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Estrogen, Endotoxin, and Alcohol-Induced Liver Injury

Also See
Alcohol Consumption – Estrogen and Progesterone In Women
How does estrogen enhance endotoxin toxicity? Let me count the ways.
PUFA and Liver Toxicity; Protection by Saturated Fats
Endotoxin: Poisoning from the Inside Out
Estrogen and Liver Toxicity
Single Bout of Binge Drinking Linked to Immune System Effects

Estrogen makes the toxic-mediator-producing cells in the liver (Kupffer cells) hypersensitive to LPS–15 times more sensitive than normal (Ikejima, et al., 1998). One way estrogen increases the toxicity of endotoxin is probably by making the intestine more permeable (Enomoto, et al., 1999). -Ray Peat, PhD

AJP – GI April 1, 1998 vol. 274 no. 4 G669-G676
Estrogen increases sensitivity of hepatic Kupffer cells to endotoxin
Kenichi Ikejima1,2, Nobuyuki Enomoto1, Yuji Iimuro1, Ayako Ikejima2, Dawn Fang1, Juliana Xu1, Donald T. Forman3, David A. Brenner2, and Ronald G. Thurman1
The relationship among gender, lipopolysaccharide (LPS), and liver disease is complex. Accordingly, the effect of estrogen on activation of Kupffer cells by endotoxin was studied. All rats given estrogen intraperitoneally 24 h before an injection of a sublethal dose of LPS (5 mg/kg) died within 24 h, whereas none of the control rats died. Mortality was prevented totally by pretreatment with gadolinium chloride, a Kupffer cell toxicant. Peak serum tumor necrosis factor-α (TNF-α) values as well as TNF-α mRNA in the liver after LPS were twice as high in the estrogen-treated group as in the untreated controls. Plasma nitrite levels and inducible nitric oxide synthase in the liver were also elevated significantly in estrogen-treated rats 6 h after LPS. Furthermore, Kupffer cells isolated from estrogen-treated rats produced about twice as much TNF-α and nitrite as controls did in response to LPS. In addition, Kupffer cells from estrogen-treated rats required 15-fold lower amounts of LPS to increase intracellular Ca2+ than controls did, and Kupffer cells from estrogen-treated animals expressed more CD14, the receptor for LPS/LPS binding protein, than controls. Moreover, estrogen treatment increased LPS binding protein mRNA dramatically in liver in 6–24 h. It is concluded that estrogen treatment in vivo sensitizes Kupffer cells to LPS, leading to increased toxic mediator production by the liver.

Am J Physiol. 1999 Sep;277(3 Pt 1):G671-7.
Estriol sensitizes rat Kupffer cells via gut-derived endotoxin.
Enomoto N, Yamashina S, Schemmer P, Rivera CA, Bradford BU, Enomoto A, Brenner DA, Thurman RG.
The relationship between gender and alcohol-induced liver disease is complex; however, endotoxin is most likely involved. Recently, it was reported that estriol activated Kupffer cells by upregulation of the endotoxin receptor CD14. Therefore, the purpose of this work was to study how estriol sensitizes Kupffer cells. Rats were given estriol (20 mg/kg ip), and Kupffer cells were isolated 24 h later. After addition of lipopolysaccharide (LPS), intracellular Ca2+ concentration was measured using a microspectrofluorometer with the fluorescent indicator fura 2, and tumor necrosis factor-alpha was measured by ELISA. CD14 was evaluated by Western analysis. One-half of the rats given estriol intraperitoneally 24 h before an injection of a sublethal dose of LPS (5 mg/kg) died within 24 h, whereas none of the control rats died. Mortality was prevented totally by sterilization of the gut with antibiotics. A similar pattern was obtained with liver histology and serum transaminases. Translocation of horseradish peroxidase was increased about threefold in gut segments by treatment with estriol. This increase was not altered by treatment with nonabsorbable antibiotics. On the other hand, endotoxin levels were increased to 60-70 pg/ml in plasma of rats treated with estriol. As expected, this increase was prevented (<20 pg/ml) by antibiotics. In isolated Kupffer cells, LPS-induced increases in intracellular Ca2+ concentration, tumor necrosis factor-alpha production, and CD14 were increased, as previously reported. All these phenomena were blocked by antibiotics. Therefore, it is concluded that estriol treatment in vivo sensitizes Kupffer cells to LPS via mechanisms dependent on increases in CD14. This is most likely due to elevated portal blood endotoxin caused by increased gut permeability.

Hepatology. 2000 Jan;31(1):117-23.
Estrogen is involved in early alcohol-induced liver injury in a rat enteral feeding model.
Yin M, Ikejima K, Wheeler MD, Bradford BU, Seabra V, Forman DT, Sato N, Thurman
The aim of this study was to investigate whether reduction in blood estrogen by removal of the ovaries would decrease the sensitivity of female rats to early alcohol-induced liver injury using an enteral ethanol feeding model, and if so, whether estrogen replacement would compensate. Livers from ovariectomized rats with or without estrogen replacement after 4 weeks of continuous ethanol exposure were compared with nonovariectomized rats in the presence or absence of ethanol. Ethanol increased serum alanine transaminase (ALT) levels from 30 +/- 6 to 64 +/- 7 U/L. This effect was blocked by ovariectomy (31 +/- 7) and totally reversed by estrogen replacement (110 +/- 23). Ethanol increased liver weight and fat accumulation, an effect that was minimized by ovariectomy and reversed partially by estrogen replacement. Infiltrating leukocytes were increased 6. 7-fold by ethanol, an effect that was blunted significantly by ovariectomy and reversed by estrogen replacement. Likewise, a similar pattern of changes was observed in the number of necrotic hepatocytes. Blood endotoxin and hepatic levels of CD14 messenger RNA (mRNA) and protein were increased by ethanol. This effect was blocked in ovariectomized rats and elevated by estrogen replacement. Moreover, Kupffer cells isolated from ethanol-treated rats with estrogen replacement produced more tumor necrosis factor alpha (TNF-alpha) than those from control and ovariectomized rats. It is concluded, therefore, that the sensitivity of rat liver to alcohol-induced injury is directly related to estrogen, which increases endotoxin in the blood and CD14 expression in the liver, leading to increased TNF-alpha production.

Am J Physiol Gastrointest Liver Physiol. 2000 Apr;278(4):G652-61.
Gender differences in early alcohol-induced liver injury: role of CD14, NF-kappaB, and TNF-alpha.
Kono H, Wheeler MD, Rusyn I, Lin M, Seabra V, Rivera CA, Bradford BU, Forman DT, Thurman RG.
The purpose of this study was to determine whether early alcohol-induced liver injury (ALI) in females is associated with changes in CD14 on Kupffer cells, activation of hepatic nuclear factor (NF)-kappaB, and expression of tumor necrosis factor (TNF)-alpha mRNA. Male and female rats were given high-fat control or ethanol-containing diets for 4 wk using the intragastric enteral protocol. Physiological parameters were similar in both genders. Ethanol was increased as tolerance developed with higher blood levels than previously observed, resulting in a fourfold increase in aspartate aminotransferase (males 389 +/- 47 IU/l vs. females 727 +/- 66 IU/l). Hepatic pathology developed more rapidly and was nearly twofold greater and endotoxin levels were significantly higher in females after ethanol. Also, expression of CD14 on Kupffer cells was 1.5-fold greater and binding of transcription factor NF-kappaB in hepatic nuclear extracts and TNF-alpha mRNA expression were threefold greater in females. These data are consistent with the hypothesis that elevated endotoxin after ethanol triggers more activation of Kupffer cells via enhanced CD14 expression in females. NF-kappaB is activated in this process, leading to increases in TNF-alpha mRNA expression in the liver and more severe liver injury in females. It is concluded that gender differences in ALI are dependent on endotoxin and a signaling cascade leading to TNF-alpha.

World J Gastroenterol. 2010 Mar 21;16(11):1377-84.
Alcoholic liver injury: influence of gender and hormones.
Eagon PK.
This article discusses several subjects pertinent to a consideration of the role of gender and hormones in alcoholic liver injury (ALI). Beginning with an overview of factors involved in the pathogenesis of ALI, we review changes in sex hormone metabolism resulting from alcohol ingestion, summarize research that points to estrogen as a cofactor in ALI, consider evidence that gut injury is linked to liver injury in the setting of alcohol, and briefly review the limited evidence regarding sex hormones and gut barrier function. In both women and female animals, most studies reveal a propensity toward greater alcohol-induced liver injury due to female gender, although exact hormonal influences are not yet understood. Thus, women and their physicians should be alert to the dangers of excess alcohol consumption and the increased potential for liver injury in females.

Am J Physiol Gastrointest Liver Physiol. 2001 Dec;281(6):G1348-56.
Increased severity of alcoholic liver injury in female rats: role of oxidative stress, endotoxin, and chemokines.
Nanji AA, Jokelainen K, Fotouhinia M, Rahemtulla A, Thomas P, Tipoe GL, Su GL, Dannenberg AJ.
Alcoholic liver injury is more severe and rapidly developing in women than men. To evaluate the reason(s) for these gender-related differences, we determined whether pathogenic mechanisms important in alcoholic liver injury in male rats were further upregulated in female rats. Male and age-matched female rats (7/group) were fed ethanol and a diet containing fish oil for 4 wk by intragastric infusion. Dextrose isocalorically replaced ethanol in control rats. We analyzed liver histopathology, lipid peroxidation, cytochrome P-450 (CYP)2E1 activity, nonheme iron, endotoxin, nuclear factor-kappa B (NF-kappa B) activation, and mRNA levels of cyclooxygenase-1 (COX-1) and COX-2, tumor necrosis factor-alpha (TNF-alpha), monocyte chemotactic protein-1 (MCP-1), and macrophage inflammatory protein-2 (MIP-2). Alcohol-induced liver injury was more severe in female vs. male rats. Female rats had higher endotoxin, lipid peroxidation, and nonheme iron levels and increased NF-kappa B activation and upregulation of the chemokines MCP-1 and MIP-2. CYP2E1 activity and TNF-alpha and COX-2 levels were similar in male and female rats. Remarkably, female rats fed fish oil and dextrose also showed necrosis and inflammation. Our findings in ethanol-fed rats suggest that increased endotoxemia and lipid peroxidation in females stimulate NF-kappa B activation and chemokine production, enhancing liver injury. TNF-alpha and COX-2 upregulation are probably important in causing liver injury but do not explain gender-related differences.

J Hepatol. 2001 Jul;35(1):46-52.
The antiestrogen toremifene protects against alcoholic liver injury in female rats.
Järveläinen HA, Lukkari TA, Heinaro S, Sippel H, Lindros KO.
BACKGROUND/AIMS:
Females are generally considered to be more susceptible to alcohol-induced liver injury than males. To elucidate whether gonadal hormones are involved, female rats were chronically treated with ethanol and with an antiestrogen.
METHODS:
Ethanol was administered in a low-carbohydrate liquid diet. Estrogen action was blocked by daily intubation of toremifene, a non-hepatotoxic second generation estrogen receptor antagonist.
RESULTS:
The female rats consuming intoxicating amounts of ethanol diet for 6 weeks developed massive microvesicular/macrovesicular steatosis, frequent inflammatory foci and spotty necrosis. Serum alanine aminotransferase increased 7-fold. Toremifene treatment did not affect steatosis, but significantly reduced inflammation and necrosis. Ethanol increased the expression of CD14 and tumor necrosis factor- (TNF) alpha mRNA and also the production of TNF-alpha by isolated Kupffer cells, but toremifene had no significant counteracting effect. However, toremifene significantly alleviated both ethanol induction of the pro-oxidant enzyme CYP2E1 and ethanol reduction of the oxidant-protective enzyme Se-glutathione peroxidase.
CONCLUSIONS:
The partial protection by toremifene against ethanol-induced liver lesions suggests a pathogenic contribution of estrogens, possibly associated with an oxygen radical mediated mechanism.

Am J Physiol. 1997 May;272(5 Pt 1):G1186-94.
Female rats exhibit greater susceptibility to early alcohol-induced liver injury than males.
Iimuro Y, Frankenberg MV, Arteel GE, Bradford BU, Wall CA, Thurman RG.
It is known that women develop hepatic injury more rapidly and with exposure to less ethanol than men; however, mechanisms remain unclear. The purpose of this study was to determine if an enteral alcohol delivery model could be used to study susceptibility of females to alcohol-induced liver injury. Male and female Wistar rats (age- or weight-matched) were given ethanol (11-12 g.kg-1.day-1) continuously for up to 4 wk via intragastric feeding, and control rats received a high-fat diet without ethanol. There were no significant differences in body weight among the groups studied. Furthermore, mean ethanol concentrations, their cyclic pattern in urine, and rates of ethanol elimination were also not different between the genders under these conditions. Ethanol treatment elevated serum aspartate aminotransferase levels in male rats to 126 +/- 10 IU/l after 4 wk. In females, however, values increased more rapidly and reached significantly higher values at 4 wk (168 +/- 18 IU/l). Steatosis, inflammation, and necrosis assessed histologically also developed more rapidly and were more severe in females than males. Steatosis due to ethanol exposure, which was localized in centrilobular areas in males, was panlobular in the female. Moreover, endotoxin in plasma, intercellular adhesion molecule 1 expression in hepatic sinusoidal-lining cells, and the number of infiltrating inflammatory cells in the liver were 2-2.5-fold greater in females than males. These changes possibly account for increased hepatic injury due to ethanol in the female.

Can J Gastroenterol. 2000 Nov;14 Suppl D:129D-135D.
Sex-related liver injury due to alcohol involves activation of Kupffer cells by endotoxin.
Thurman RG.
Females have a greater susceptibility to ethanol-induced liver injury than males. Females who drink ethanol regularly and have been overweight for 10 years or more are at greater risk for both hepatitis and cirrhosis than males, and females develop ethanol-induced liver injury more rapidly and with less ethanol than males. Female rats on an enteral ethanol protocol exhibit injury more quickly than males and have widespread fatty changes over a larger portion of the liver lobule. Moreover, levels of plasma endotoxin, intracellular adhesion molecule-1, free radical adducts, infiltrating neutrophils and nuclear factor kappa B are doubled in female rat livers compared with male rat livers after enteral ethanol treatment. Additionally, estrogen treatment in vivo increases the sensitivity of hepatic macrophages or Kupffer cells to endotoxin. Evidence has been presented that Kupffer cells are pivotal in the development of ethanol-induced liver injury. Destroying Kupffer cells with gadolinium chloride or decreasing bacterial endotoxin by sterilizing the gut with antibiotics inhibits early inflammation due to ethanol. Similar results have been obtained with anti-tumour necrosis factor-alpha antibody. These data pointed to the hypothesis that ethanol-induced liver injury involves elevations in circulating endotoxin concentrations leading to activation of Kupffer cells, which causes a hypoxia-reoxygenation injury. This theory has been tested using pimonidazole, a 2-nitroimidazole marker, to quantify hypoxia in downstream, pericentral regions of the hepatic lobule. After chronic enteral ethanol treatment, pimonidazole binding doubles. Enteral ethanol also increases free radicals detected with electron spin resonance. Radical adducts, with coupling constants such as alpha-hydroxyethyl radical, have been shown to arise from ethanol. Importantly, hypoxia and radical production detected in bile are also decreased by the destruction of Kupffer cells with gadolinium chloride. These data support the hypothesis that Kupffer cells contribute to the vital sex differences in liver injury caused by ethanol.

J Gastroenterol Hepatol. 1998 Sep;13 Suppl:S39-50.
The role of gut-derived bacterial toxins and free radicals in alcohol-induced liver injury.
Thurman RG, Bradford BU, Iimuro Y, Knecht KT, Arteel GE, Yin M, Connor HD, Wall C, Raleigh JA, Frankenberg MV, Adachi Y, Forman DT, Brenner D, Kadiiska M, Mason RP.
Previous research from this laboratory using a continuous enteral ethanol (EtOH) administration model demonstrated that Kupffer cells are pivotal in the development of EtOH-induced liver injury. When Kupffer cells were destroyed using gadolinium chloride (GdCl3) or the gut was sterilized with polymyxin B and neomycin, early inflammation due to EtOH was blocked. Anti-tumour necrosis factor (TNF)-alpha antibody markedly decreased EtOH-induced liver injury and increased TNF-mRNA. These findings led to the hypothesis that EtOH-induced liver injury involves increases in circulating endotoxin leading to activation of Kupffer cells. Pimonidazole, a nitro-imidazole marker, was used to detect hypoxia in downstream pericentral regions of the lobule. Following one large dose of EtOH or chronic enteral EtOH for 1 month, pimonidazole binding was increased significantly in pericentral regions of the liver lobule, which was diminished with GdCl3. Enteral EtOH increased free radical generation detected with electron spin resonance (ESR). These radical species had coupling constants matching alpha-hydroxyethyl radical and were shown conclusively to arise from EtOH based on a doubling of the ESR lines when 13C-EtOH was given. Alpha-hydroxyethyl radical production was also blocked by the destruction of Kupffer cells with GdCl3. It is known that females develop more severe EtOH-induced liver injury more rapidly and with less EtOH than males. Female rats on the enteral protocol exhibited more rapid injury and more widespread fatty changes over a larger portion of the liver lobule than males. Plasma endotoxin, ICAM-1, free radical adducts, infiltrating neutrophils and transcription factor NFkappaB were approximately two-fold greater in livers from females than males after 4 weeks of enteral EtOH treatment. Furthermore, oestrogen treatment increased the sensitivity of Kupffer cells to endotoxin. These data are consistent with the hypothesis that Kupffer cells participate in important gender differences in liver injury caused by ethanol.

Journal of Hepatology 35 (2001) 130±133
Alcoholic liver disease: a matter of hormones?
Han Moshage

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Childhood conditions influence adult progesterone levels

Also see:
Nutrition and Brain Growth in Chick Embryos
PUFA, Estrogen, Obesity and Early Onset of Puberty

Quotes by Ray Peat, PhD:
“It seems clear that the course of degenerative aging processes is set in young adulthood (or even earlier), and that it is never too early to be concerned with correcting processes that are going in the wrong direction. (See Walker, et al., 1988, and Smith, et al., 1992.)

In “The Biological Generality of Progesterone” (1979) I proposed that the life-long trajectory of energy production and longevity was strongly influenced by prenatal nutrition and progesterone. This idea was based on work by people such as Marion Diamond, who showed that prenatal progesterone enlarges the cortex of the brain, and that estrogen makes it smaller, and Leonell Strong, who showed that a treatment that lowered the estrogen function in a young mouse could produce cancer-free offspring for several generations. Strong’s work was very encouraging, because it showed that biological problems that had been “bred in” over many generations could be corrected by some simple metabolic treatments.”

“Many factors, including poor nutrition, climate, emotional or physical stress (even excessive running) and toxins, can cause a progesterone deficiency. Use of estrogens, birth control pills and even IUDs can also bring about a deficiency. Animal studies and clinical experience suggests that the prenatal hormonal environment (a mother’s excess of estrogen during pregnancy) can incline a person toward a deficiency of progesterone relative to estrogen.”

PLoS Med. 2007 May;4(5):e167.
Childhood conditions influence adult progesterone levels.
Núñez-de la Mora A, Chatterton RT, Choudhury OA, Napolitano DA, Bentley GR.
BACKGROUND:
Average profiles of salivary progesterone in women vary significantly at the inter- and intrapopulation level as a function of age and acute energetic conditions related to energy intake, energy expenditure, or a combination of both. In addition to acute stressors, baseline progesterone levels differ among populations. The causes of such chronic differences are not well understood, but it has been hypothesised that they may result from varying tempos of growth and maturation and, by implication, from diverse environmental conditions encountered during childhood and adolescence.
METHODS AND FINDINGS:
To test this hypothesis, we conducted a migrant study among first- and second-generation Bangladeshi women aged 19-39 who migrated to London, UK at different points in the life-course, women still resident in Bangladesh, and women of European descent living in neighbourhoods similar to those of the migrants in London (total n = 227). Data collected included saliva samples for radioimmunoassay of progesterone, anthropometrics, and information from questionnaires on diet, lifestyle, and health. Results from multiple linear regression, controlled for anthropometric and reproductive variables, show that women who spend their childhood in conditions of low energy expenditure, stable energy intake, good sanitation, low immune challenges, and good health care in the UK have up to 103% higher levels of salivary progesterone and an earlier maturation than women who develop in less optimal conditions in Sylhet, Bangladesh (F9,178 = 5.05, p < 0.001, standard error of the mean = 0.32; adjusted R(2) = 0.16). Our results point to the period prior to puberty as a sensitive phase when changes in environmental conditions positively impact developmental tempos such as menarcheal age (F2,81 = 3.21, p = 0.03) and patterns of ovarian function as measured using salivary progesterone (F2,81 = 3.14, p = 0.04).
CONCLUSIONS:
This research demonstrates that human females use an extended period of the life cycle prior to reproductive maturation to monitor their environment and to modulate reproductive steroid levels in accordance with projected conditions they might encounter as adults. Given the prolonged investment of human pregnancy and lactation, such plasticity (extending beyond any intrauterine programming) enables a more flexible and finely tuned adjustment to the potential constraints or opportunities of the later adult environment. This research is the first, to our knowledge, to demonstrate a postuterine developmental component to variation in reproductive steroid levels in women.

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Exercise Induced Menstrual Disorders

Also See:
Exercise Induced Stress
Exercise and Effect on Thyroid Hormone
Potential Adverse Cardiovascular Effects from Excessive Endurance Exercise
Ray Peat, PhD: Quotes Relating to Exercise
Ray Peat, PhD and Concentric Exercise

“Sometimes progesterone seems to be chronically deficienct (leading to slight-though possibly prolonged-menstruation, or amenorrhea), in women who exercise hard. Since progesterone can be converted into cortisone to handle stress, this would explain why well trained athletes (who need lots of cortisone) so often miss periods. It seems to be a simple over-consumption of progesterone, which is probably a reasonable biological adaptation, preventing pregnancy during times of stress.” -Ray Peat, PhD

Proc Biol Sci. 1998 October 7; 265(1408): 1847–1851.
Physical work causes suppression of ovarian function in women.
G Jasieńska and P T Ellison
The suppression of reproductive function is known to occur in women engaging in activities that require high energetic expenses, such as sport participation and subsistence work. It is still unclear, however, if reproductive suppression is a response to high levels of energy expenditure, or only to the resulting state of negative energy balance. To our knowledge, this study provides the first evidence that work-related energy expenditure alone, without associated negative energy balance, can lead to the suppression of reproductive function in women. We document suppression of ovarian function expressed as lowered salivary progesterone levels in women from an agricultural community who work hard, but remain in neutral energy balance. We propose two alternative evolutionary explanations (the ‘pre-emptive ovarian suppression’ hypothesis and the ‘constrained down-regulation’ hypothesis) for the observed results.

Croat Med J. 2001 Feb;42(1):79-82.
Influence of high intensity training on menstrual cycle disorders in athletes.
Dusek T.
AIM:
To estimate the influence of intensive training on menstrual cycles in female athletes.
METHOD:
The questionnaire was used to determine the time of menarche, and the prevalence of primary and secondary amenorrhea and dysmenorrhea in 72 active female athletes from Zagreb (10 volleyball players, 18 basketball players, 10 ballet dancers, and 34 runners) aged between 15 and 21. The control group comprised 96 girls of the same age not engaged in any sports activity.
RESULTS:
The prevalence of secondary amenorrhea was three times higher in athletes than in the control group (p=0.037). The prevalence of primary amenorrhea was substantially higher in athletes than in the control group (6/72 vs. 0/96, p=0.014), whereas the prevalence of dysmenorrhea was twofold lower in athletes than in the control group (p<0.001). The highest prevalence of secondary amenorrhea was recorded in runners (14/31), particularly long-distance runners (11/17), whereas there was only one case of secondary amenorrhea among basketball players. Menarche was significantly delayed in the athletes who started physical activities before the onset of menstruation (13.8+1.4 vs. 12.6+1.0 years, p<0.001).
CONCLUSION:
High-intensity training before menarche postpones its onset. Type of training may be related to a significantly higher prevalence of secondary amenorrhea in runners than in basketball players.

Br J Sports Med. 2005 Mar;39(3):141-7.
Participation in leanness sports but not training volume is associated with menstrual dysfunction: a national survey of 1276 elite athletes and controls.
Torstveit MK, Sundgot-Borgen J.
OBJECTIVE:
To examine the prevalence of menstrual dysfunction in the total population of Norwegian elite female athletes and national representative controls in the same age group.
METHODS:
A detailed questionnaire that included questions on training and/or physical activity patterns, menstrual, dietary, and weight history, oral contraceptive use, and eating disorder inventory subtests was administered to all elite female athletes representing the country at the junior or senior level (aged 13-39 years, n = 938) and national representative controls in the same age group (n = 900). After exclusion, a total of 669 athletes (88.3%) and 607 controls (70.2%) completed the questionnaire satisfactorily.
RESULTS:
Age at menarche was significantly (p<0.001) later in athletes (13.4 (1.4) years) than in controls (13.0 (1.3) years), and differed among sport groups. A higher percentage of athletes (7.3%) than controls (2.0%) reported a history of primary amenorrhoea (p<0.001). A similar percentage of athletes (16.5%) and controls (15.2%) reported present menstrual dysfunction, but a higher percentage of athletes competing in leanness sports reported present menstrual dysfunction (24.8%) than athletes competing in non-leanness sports (13.1%) (p<0.01) and controls (p<0.05).
CONCLUSIONS:
These novel data include virtually all eligible elite athletes, and thus substantially extend previous studies. Age at menarche occurred later and the prevalence of primary amenorrhoea was higher in elite athletes than in controls. A higher percentage of athletes competing in sports that emphasise thinness and/or a specific weight reported present menstrual dysfunction than athletes competing in sports focusing less on such factors and controls. On the basis of a comparison with a previous study, the prevalence of menstrual dysfunction was lower in 2003 than in 1993.

Hum Reprod. 2010 Feb;25(2):491-503. Epub 2009 Nov 26.
High prevalence of subtle and severe menstrual disturbances in exercising women: confirmation using daily hormone measures.
De Souza MJ, Toombs RJ, Scheid JL, O’Donnell E, West SL, Williams NI.
BACKGROUND:
The identification of subtle menstrual cycle disturbances requires daily hormone assessments. In contrast, the identification of severe menstrual disturbances, such as amenorrhea and oligomenorrhea, can be established by clinical observation. The primary purpose of this study was to determine the frequency of subtle menstrual disturbances, defined as luteal phase defects (LPD) or anovulation, in exercising women, with menstrual cycles of 26-35 days, who engage in a variety of sports, both recreational and competitive. Secondly, the prevalence of oligomenorrhea and amenorrhea was also determined via measurement of daily urinary ovarian steroids rather than self report alone.
METHODS:
Menstrual status was documented by daily measurements of estrone and pregnanediol glucuronide and luteinizing hormone across two to three consecutive cycles and subsequently categorized as ovulatory (Ovul), LPD, anovulatory (Anov), oligomenorrheic (Oligo) and amenorrheic (Amen) in sedentary (Sed) and exercising (Ex) women.
RESULTS:
Sed (n = 20) and Ex women (n = 67) were of similar (P > 0.05) age (26.3 +/- 0.8 years), weight (59.3 +/- 1.8 kg), body mass index (22.0 +/- 0.6 kg/m2), age of menarche (12.8 +/- 0.3 years) and gynecological maturity (13.4 +/- 0.9 years). The Sed group exercised less (P < 0.001) (96.7 +/- 39.1 versus 457.1 +/- 30.5 min/week) and had a lower peak oxygen uptake (34.4 +/- 1.4 versus 44.3 +/- 0.6 ml/kg/min) than the Ex group. Among the menstrual cycles studied in the Sed group, the prevalence of subtle menstrual disturbances was only 4.2% (2/48); 95.8% (46/48) of the observed menstrual cycles were ovulatory. This finding stands in stark contrast to that observed in the Ex group where only 50% (60/120) of the observed menstrual cycles were ovulatory and as many as 50% (60/120) were abnormal. Of the abnormal cycles in the Ex group, 29.2% (35/120) were classified as LPD (short, inadequate or both) and 20.8% (25/120) were classified as Anov. Among the cycles of Ex women with severe menstrual disturbances, 3.5% (3/86) of the cycles were Oligo and 33.7% (29/86) were Amen. No cycles of Sed women (0/20) displayed either Oligo or Amen.
CONCLUSIONS:
This study suggests that approximately half of exercising women experience subtle menstrual disturbances, i.e. LPD and anovulation, and that one third of exercising women may be amenorrheic. Estimates of the prevalence of subtle menstrual disturbances in exercising women determined by the presence or absence of short or long cycles does not identify these disturbances. In light of known clinical consequences of menstrual disturbances, these findings underscore the lack of reliability of normal menstrual intervals and self report to infer menstrual status.

N Engl J Med. 1985 May 23;312(21):1349-53.
Induction of menstrual disorders by strenuous exercise in untrained women.
Bullen BA, Skrinar GS, Beitins IZ, von Mering G, Turnbull BA, McArthur JW.
We performed a prospective study of 28 initially untrained college women with documented ovulation and luteal adequacy to determine whether strenuous exercise spanning two menstrual cycles would induce menstrual disorders. To ascertain the influence, if any, that weight loss might exert, we randomly assigned the subjects to weight-loss and weight-maintenance groups. Subjects were expected to run 4 miles (6.4 km) per day, progressing to 10 miles (16.1 km) per day by the fifth week, and to engage daily in 3 1/2 hours of moderate-intensity sports. The normalcy of the menstrual cycles during the period of exercise was judged independently according to clinical and hormonal criteria, the latter comprising serial measurements of gonadotropin and sex-steroid excretion. A higher percentage of abnormalities proved to be detectable by hormonal means (P less than 0.02). Only four subjects (three in the weight-maintenance group) had a normal menstrual cycle during training. In the weight-loss group, the number of women who had luteal abnormalities as compared with those who lost the surge in luteinizing hormone altered significantly over time, the latter occurring more frequently (P less than 0.01) as training progressed. Within six months of termination of the study, all subjects were again experiencing normal menstrual cycles. We conclude that vigorous exercise, particularly if compounded by weight loss, can reversibly disturb reproductive function in women.

J Clin Endocrinol Metab. 1991 Jun;72(6):1350-8.
Exercise induces two types of human luteal dysfunction: confirmation by urinary free progesterone.
Beitins IZ, McArthur JW, Turnbull BA, Skrinar GS, Bullen BA.
We have previously reported that during 2 months of strenuous exercise, untrained young women with documented ovulatory menstrual cycles developed secondary oligoamenorrhea and luteal phase defects. In this study we tested the hypothesis that such abnormalities arise by altered neuroendocrine regulation of menstrual hormone secretion and that weight loss potentiates such effects. We supply a detailed analysis of the 20 cycles, of the total of 53, in which luteal phase abnormalities occurred. During the control month and 2 exercise months, all subjects collected daily overnight urine samples for the determination of LH, FSH, estriol (E3), and free progesterone (P) excretion by RIAs and creatinine by chemical assay. The characteristics of the abnormal luteal phase cycles were determined by comparing the excreted hormone levels and patterns during the control cycles with those of exercise cycles. The area under the curve (AUC) for each hormone was calculated for the follicular and luteal phases of each cycle. Six of the exercise cycles exhibited an inadequate luteal phase. This was characterized by a mean integrated P area of 202.4 (SEM, -61.8) nmol/day.nmol creatinine, compared with 331.7 (SEM, 64.7) during the corresponding control cycles, over a period of 9 or more days after the urinary LH peak to the onset of menses. Fourteen of the exercise cycles exhibited a short luteal phase. This was characterized by a mean integrated P area of 75.9 (30.9) nmol/day.nmol creatinine, compared to 267 (61.7) during the corresponding control cycles, over a span of 8 days or less from the urinary LH peak to the onset of menses. Additional abnormalities occurred only in the short luteal phase cycles. These included an increase in the length and AUC for E3 of the follicular phase and a decrease in the AUC of LH during the luteal phase. We conclude that the initiation of strenuous endurance training in previously ovulating untrained women frequently leads to corpus luteum dysfunction associated with insufficient P secretion and, in the case of short luteal phase cycles, decreased luteal phase length. That exercise may alter the neuroendocrine system is suggested by a delay in the ovulatory LH peak in spite of increased E3 excretion; moreover, less LH is excreted during the luteal phase. The lack of positive feedback to estrogens and decreased LH secretion during the luteal phase could compromise corpus luteum function. In contrast, decreased free P excretion was the sole abnormality noted in menstrual cycles with an inadequate luteal phase.

Annu Rev Med. 1988;39:443-51.
Exercise-induced menstrual dysfunction.
Henley K, Vaitukaitis JL.
Menstrual cycle changes associated with vigorous exercise can range widely. They may be only subtle abnormalities, ranging from delayed onset of spontaneous menses or anovulatory cycles to loss of spontaneous menses. They may be more serious, however. Significant adverse bone mineral changes, resulting in clinically significant osteoporosis and fractures, may occur concomitantly with exercise-induced menstrual dysfunction.

Sports Med. 1990 Oct;10(4):218-35.
Physical exercise and menstrual cycle alterations. What are the mechanisms?
Keizer HA, Rogol AD.
The prevalence of menstrual cycle alterations in athletes is considerably higher than in sedentary controls. There appears to be a multicausal aetiology, which makes it extremely difficult to dissociate the effects of physical exercise on the menstrual cycle from the other predisposing factors. From cross-sectional studies it appeared that physical training eventually might lead to shortening of the luteal phase and secondary amenorrhoea. Prospective studies in both trained and previously untrained women have shown that the amount and/or the intensity of exercise has to exceed a certain limit in order to elicit this phenomenon. We hypothesise, therefore, that apart from a certain predisposition, athletes with a training-induced altered menstrual cycle are overreached (short term overtraining, which is reversible in days to weeks after training reduction). Menstrual cycle alterations are most likely caused by subtle changes in the episodic secretion pattern of luteinising hormone (LH) as have been found in sedentary women with hypothalamic amenorrhoea as well as in athletes after very demanding training. The altered LH secretion then, might be caused by an increased corticotrophin-releasing hormone (CRH) secretion which inhibits the gonadotrophin-releasing hormone (GnRH) release. In addition, increased CRH tone will lead to increased beta-endorphin levels which will also inhibit the GnRH signaller. Finally, the continuous activation of the adrenals will result in a higher catecholamine production, which may be converted to catecholestrogens. These compounds are known to be potent inhibitors of GnRH secretion. In conclusion, menstrual cycle alterations are likely to occur after very demanding training, which causes an increase secretion of antireproductive hormones. These hormones can inhibit the normal pulsatile secretion pattern of the gonadotrophins.

Med Sci Sports Exerc. 2003 Sep;35(9):1553-63.
Menstrual disturbances in athletes: a focus on luteal phase defects.
De Souza MJ.
Subtle menstrual disturbances that affect the largest proportion of physically active women and athletes include luteal phase defects (LPD). Disorders of the luteal phase, characterized by poor endometrial maturation as a result of inadequate progesterone (P4) production and short luteal phases, are associated with infertility and habitual spontaneous abortions. In recreational athletes, the 3-month sample prevalence and incidence rate of LPD and anovulatory menstrual cycles is 48% and 79%, respectively. A high proportion of active women present with LPD cycles in an intermittent and inconsistent manner. These LPD cycles are characterized by reduced follicle-stimulating hormone (FSH) during the luteal-follicular transition, a somewhat blunted luteinizing hormone surge, decreased early follicular phase estradiol excretion, and decreased luteal phase P4 excretion both with and without a shortened luteal phase. LPD cycles in active women are associated with a metabolic hormone profile indicative of a hypometabolic state that is similar to that observed in amenorrheic athletes but not as comprehensive or severe. These metabolic alterations include decreased serum total triiodothyronine (T3), leptin, and insulin levels. Bone mineral density in these women is apparently not reduced, provided an adequate estradiol environment is maintained despite decreased P4. The high prevalence of LPD warrants further investigation to assess health risks and preventive strategies.

J Clin Endocrinol Metab. 2003 Jan;88(1):337-46.
Luteal phase deficiency in recreational runners: evidence for a hypometabolic state.
De Souza MJ, Van Heest J, Demers LM, Lasley BL.
Exercising women with amenorrhea exhibit a hypometabolic state. The purpose of this study was to evaluate the relationship of luteal phase deficient (LPD) menstrual cycles to metabolic hormones, including thyroid, insulin, human GH (hGH), leptin, and IGF-I and its binding protein levels in recreational runners. Menstrual cycle status was determined for three consecutive cycles in sedentary and moderately active women. Menstrual status was defined as ovulatory or LPD. Subjects were either sedentary (n = 10) or moderately active (n = 20) and were matched for age (27.7 +/- 1.2 yr), body mass (60.2 +/- 3.3 kg), menstrual cycle length (28.4 +/- 0.9 d), and reproductive age (14.4 +/- 1.2 yr). Daily urine samples for the determination of estrone conjugates, pregnanediol 3-glucuronide, and urinary levels of LH were collected. Blood was collected on a single day during the follicular phase (d 2-6) of each menstrual cycle for analysis of TSH, insulin, total T3, total T4, free T4, leptin, hGH, IGF-I, and IGF binding protein (IGFBP)-1 and IGFBP-3. Among the 10 sedentary subjects, 28 of 31 menstrual cycles were categorized as ovulatory (SedOvul). Among the 20 exercising subjects, 24 menstrual cycles were included in the ovulatory category (ExOvul), and 21 menstrual cycles were included in the LPD category (ExLPD). TSH, total T4, and free T4 levels were not significantly different among the three categories of cycles. Total T3 was suppressed (P = 0.035) in the ExLPD (1.63 +/- 0.07 nmol/liter) and the ExOvul categories of cycles (1.75 +/- 0.8 nmol/liter) compared with the SedOvul category of cycles (2.15 +/- 0.1 nmol/liter). Leptin levels were lower (P < 0.001) in both the ExOvul (5.2 +/- 0.4 microg/liter) and the ExLPD categories of cycles (5.1 +/- 0.4 microg/liter) when compared with the SedOvul category of cycles (13.7 +/- 1.7 microg/liter). Insulin was lower (P = 0.009) only in the ExLPD category of cycles (31.9 +/- 2.8 pmol/liter) compared with the SedOvul (60.4 +/- 8.3 pmol/liter) and ExOvul (61.8 +/- 10.4 pmol/liter) categories of cycles. IGF-I, IGFBP-1, IGFBP-3, IGF-I/IGFBP-1, IGF-I/IGFBP-3, and hGH were comparable among the different categories of cycles. These data suggest that exercising women with LPD menstrual cycles exhibit hormonal alterations consistent with a hypometabolic state that is similar to that observed in amenorrheic athletes and other energy-deprived states, although not as comprehensive. These alterations may represent a metabolic adaptation to an intermittent short-term negative energy balance.

J Clin Endocrinol Metab. 1998 Dec;83(12):4220-32.
High frequency of luteal phase deficiency and anovulation in recreational women runners: blunted elevation in follicle-stimulating hormone observed during luteal-follicular transition.
De Souza MJ, Miller BE, Loucks AB, Luciano AA, Pescatello LS, Campbell CG, Lasley BL.
The purposes of this investigation were to evaluate the characteristics of three consecutive menstrual cycles and to determine the frequency of luteal phase deficiency (LPD) and anovulation in a sample of sedentary and moderately exercising, regularly menstruating women. For three consecutive menstrual cycles, subjects collected daily urine samples for analysis of FSH, estrone conjugates (E1C), pregnanediol-3-glucuronide (PdG), and creatinine (Cr). Sedentary (n=11) and exercising (n=24) groups were similar in age (27.0+/-1.3 yr), weight (60.3+/-3.1 kg), gynecological age (13.8+/-1.2 yr), and menstrual cycle length (28.3+/-0.8 days). Menstrual cycles were classified by endocrine data as ovulatory, LPD, or anovulatory. No sedentary women (0%) had inconsistent menstrual cycle classifications from cycle to cycle, but 46% of the exercising women were inconsistent. The sample prevalence of LPD in the exercising women was 48%, and the 3-month sample incidence was 79%. In the sedentary women, 90% of all menstrual cycles were ovulatory (SedOvul; n=28), whereas in the exercising women only 45% were ovulatory (ExOvul; n=30); 43% were LPD (ExLPD; n=28), and 12% were anovulatory (ExAnov; n=8). In ExLPD cycles, the follicular phase was significantly longer (17.9+/-0.7 days), and the luteal phase was significantly shorter (8.2+/-0.5 days) compared to ExOvul (14.8+/-0.9 and 12.9+/-0.3 days) and SedOvul (15.9+/-0.6 and 12.9+/-0.4 days) cycles. Luteal phase PdG excretion was lower (P < 0.001) in ExLPD (2.9+/-0.3 microg/mg Cr) and ExAnov (0.8+/-0.1 microg/mg Cr) cycles compared to SedOvul cycles (5.0+/-0.4 microg/mg Cr). ExOvul cycles also had less (P < 0.01) PdG excretion during the luteal phase (3.7+/-0.3 microg/mg Cr) than the SedOvul cycles. E1C excretion during follicular phase days 2-5 was lower (P=0.05) in ExOvul, ExLPD, and ExAnov cycles compared to SedOvul cycles and remained lower (P < 0.02) in the ExLPD and ExAnov cycles during days 6-12. The elevation in FSH during the luteal-follicular transition was lower (P < 0.007) in ExLPD (0.7+/-0.1 ng/mg Cr) cycles compared to SedOvul and ExOvul cycles (1.0+/-0.1 and 1.1+/-0.1 ng/mg Cr, respectively). Energy balance and energy availability were lower (P < 0.05) in ExAnov cycles than in other menstrual cycle categories. The blunted elevation in FSH during the luteal-follicular transition in exercising women with LPD may explain their lower follicular estradiol levels. These alterations in FSH may act in concert with disrupted LH pulsatility as a primary and proximate factor in the high frequency of luteal phase and ovulatory disturbances in regularly menstruating, exercising women.

J Clin Endocrinol Metab. 1997 Sep;82(9):2867-76.
Bone health is not affected by luteal phase abnormalities and decreased ovarian progesterone production in female runners.
De Souza MJ, Miller BE, Sequenzia LC, Luciano AA, Ulreich S, Stier S, Prestwood K, Lasley BL.
The primary purpose of this study was to determine whether decreased ovarian progesterone production, associated with short and inadequate luteal phases in exercising women, was associated with decreased bone mineral density (BMD) and altered bone metabolism. Thirty-three eumenorrheic menstruating women participated in this study for 3 months. Subjects were required to collect daily urine samples for three consecutive menstrual cycles and have blood and urine collected weekly. Daily urine samples were analyzed for free LH, estrone conjugates (E1C), and pregnanediol 3-glucuronide (PdG), adjusted for creatinine, whereas weekly blood and urine samples were analyzed for bone markers, estradiol, progesterone, FSH, and LH. Based on the analyses of these samples, subjects were divided into three groups: sedentary ovulatory (SedOvul; n = 9), exercising ovulatory (ExOvul; n = 14), and exercising luteal phase defects (ExLPD; n = 10). The three groups were matched for age (27.6 +/- 1.0 yr), weight (60.6 +/- 1.9 cm), and reproductive maturity (14.5 +/- 1.0 yr), PdG production during the luteal phase was lower (P = 0.004) in the ExLPD women compared to that in the SedOvul group (2.4 +/- 0.4 vs. 5.1 +/- 0.6 ng/mL creatinine, respectively). The ExOvul group also had less (P < 0.01) PdG production during the luteal phase (3.5 +/- 0.3 ng/mL creatinine) compared to the SedOvul group. The total production of PdG, as assessed by area under the curve analysis, was also lower (P < 0.001) in the ExOvul and ExLPD groups compared to that in the SedOvul group. E1C production, however, was not different (P > 0.05) among the groups, except for E1C during the early follicular phase, which was lower (P = 0.043) in the ExLPD group than that in the SedOvul group. BMD and biochemical markers of bone metabolism were unaffected by and not associated with the compromised progesterone environment, but BMD values at the proximal femur (r = 0.354; P = 0.061) and total body (r = 0.359; P = 0.056) were associated with decreased early follicular E1C production. We conclude the following. 1) Luteal phase disturbances occur independent of training volume, and volume of training does not have to be severe to result in menstrual disturbances. 2) As a result of exercise, disturbance in progesterone production is not associated with decreased bone mass. 3) Long follicular phases are associated with reduced estrogen production during the early follicular phase, which are both associated with decreased bone mass. 4) Provided the estradiol status is adequately maintained, BMD is unaffected by decreased progesterone production associated with short and inadequte luteal phases in exercising women.

Clin Endocrinol (Oxf). 1990 Sep;33(3):345-53.
Luteinizing hormone and follicle stimulating hormone secretion patterns in female athletes with and without menstrual disturbances.
Pirke KM, Schweiger U, Broocks A, Tuschl RJ, Laessle RG.
Thirty-one young female athletes and 13 age-matched sedentary controls were studied throughout one menstrual cycle or over a 6 week period. Blood was sampled on 5 days per week. Episodic gonadotrophin secretion was measured in the early follicular phase and in the late luteal phase by blood sampling over a 12-h period at 15-min intervals. Eight athletes had anovulatory cycles, nine had impaired progesterone (P4) secretion during the luteal phase and 14 had normal cycles as judged from oestradiol (E2) and P4 plasma levels. Athletes with normal cycles had shorter cycles, lower E2 maxima at midcycle, and lower E2 and P4 concentrations during the luteal phase than had sedentary controls. Episodic luteinizing hormone (LH) secretion in the early follicular phase was significantly impaired in the anovulatory athletes: the average LH values over 12 h and the number of secretion episodes were significantly reduced. No significant changes were seen in follicle stimulating hormone secretion.

Br J Obstet Gynaecol. 1982 Jul;89(7):507-10.
Body weight, exercise and menstrual status among ballet dancers in training.
Abraham SF, Beumont PJ, Fraser IS, Llewellyn-Jones D.
A prospective study of the menstrual pattern and weight changes was made in the first year of training of 29 new female entrants to a professional ballet school. Seventy-nine per cent of the student girls had menstrual disturbances at entry: primary amenorrhoea, four; secondary amenorrhoea, 11; irregular menses, eight. The incidence of secondary amenorrhoea increase substantially by the end of the year (20), but was not associated with any significant change in body weight. Only three students menstruated regularly during the year. Menstrual regularity improved during periods of injury and long vacation and it appears that deterioration of the menstrual pattern during dancing periods was related to strenuous physical exercise rather than to any change in body weight.

Gynecol Endocrinol. 2006 Jan;22(1):31-5.
Influence of high-intensity training and of dietetic and anthropometric factors on menstrual cycle disorders in ballet dancers.
Castelo-Branco C, Reina F, Montivero AD, Colodrón M, Vanrell JA.
Background. Intensity of exercise and low energy consumption, specific type and amount of training, early age at initiation, previous menstrual dysfunctions, low body mass index (BMI) or percentage body fat, pathological feeding habits and psychological stress have been suggested as potential factors accountable for menstrual irregularities in female athletes.
Aim. To evaluate the influence of intensive training and of dietetic and anthropometric factors on menstrual cycles in female ballet dancers.
Method. A case-control study, in which a structured interview and physical examination were carried out in two groups of teenagers aged between 12 and 18 years. The study included a total of 115 adolescent girls distributed in two groups: dancers (group B, n = 38) and girls of the same age not engaged in any sports activity (group C, n = 77).
Results. Early starting high-intensity training delayed the onset of menarche ( p < 0.001). Dancers had a higher prevalence of oligomenorrhea and amenorrhea than control girls ( p = 0.004). Additionally, the dancers had lower scores in anthropometric variables: breast circumference 80 cm vs. 86.6 cm for controls ( p = 0.0001), low weight in 18% of dancers vs. 2.6% of controls ( p = 0.0001), and low height in 18% of dancers vs. 9% of controls ( p = 0.016). In addition, in dancers, low BMI was observed in 21% compared with 13% of controls ( p = 0.0001). Finally, 32% of the dancers were on a weight-control diet while this percentage decreased to 12% for the girls in control group (odds ratio = 3.49, 95% confidence interval = 1.31-9.25).
Conclusions. In ballet dancers, high-intensity training was associated with late onset of menarche, menstrual disorders, lower weight and height development, and abnormal feeding behaviors.

Clin Obstet Gynecol. 1983 Sep;26(3):728-35.
Exercise, sports, and menstrual dysfunction.
Hale RW.
With the increasing involvement of women in exercise programs, the physician is faced with more and more questions regarding the effect of exercise upon the reproductive system. Currently, it appears that premenarchal training may have the effect of delaying the onset of menses in some girls. There is no evidence that it delays the other stage of puberty or that it causes any harmful development by this delay. In the postmenarchal woman, strenuous exercise can definitely alter her bleeding pattern. The usual result is oligomenorrhea progressing toward amenorrhea as the exercise increases. This is not a universal phenomenon, however, and other factors such as percentage of body fat, stress, diet, and energy drain also play a role. The menses will usually resume its preexercise pattern after a period of rest.

Med Clin North Am. 1985 Jan;69(1):83-95.
Causes, evaluation, and management of athletic oligo-/amenorrhea.
Shangold MM.
Oligomenorrhea and amenorrhea are more common among athletes than among the general population. Although these conditions in athletes are often related to exercise and thinness, they may be caused by serious pathology too. All athletes with menstrual dysfunction deserve thorough evaluation and most need treatment.

Aust Fam Physician. 1984 Sep;13(9):659-63.
Oligomenorrhoea and amenorrhoea associated with exercise. A literature review.
Williams M.
Increasing numbers of women are embarking on more strenuous and constant exercise; their menstrual patterns are changing as a result. This review of the literature indicates that oligomenorrhoea and amenorrhoea in the physically active (particularly distance runners, gymnasts, ballet dancers and swimmers) are related to each woman’s physiological and psychological makeup.

Am Fam Physician. 1984 May;29(5):233-7.
The female athlete.
Wilkerson LA.
Anatomic considerations are the female athlete’s wider pelvis, shorter extremities and lower center of gravity. There is little qualitative difference in the muscle tissue of men and women; differences in strength stem from the amount of muscle mass. Amenorrhea/oligomenorrhea is common in runners, ballet dancers, cyclists, gymnasts, body builders, figure skaters and, to a lesser extent, swimmers. Pregnancy limits activity, but current evidence indicates that exercise during pregnancy is not harmful to either the mother or the fetus.

Int J Neurosci. 2006 Dec;116(12):1549-63.
Effects of menstrual cycle on sports performance.
Kishali NF, Imamoglu O, Katkat D, Atan T, Akyol P.
The aim of this study was to examine the effects of menstrual cycle on female athletes’ performance. Forty-eight teak-wondo athletes, 76 judoka, 81 volleyball, and 36 basketball players (total 241) elite athletes participated in the study. A questionnaire constituted from 21 questions about menstrual cycle applied. A one-way analysis of variance and scheffe tests were performed to assess differences between sport branches about physical and physiological characteristics. Chi square was used to evaluate the regularity of menstrual cycle, performance, and drug taking. The mean age of teak-wondo athletes, judokas, volleyball and basketball players were 20.71 +/- 0.41, 16.91 +/- 0.27, 21.22 +/- 0.26, and 21.03 +/- 0.63 years, respectively. The menarche ages of the athletes were 13.92, 13.22, 13.75, 13.86 years, respectively. 27.8% participated in regional competitions, 46.1% participated in just the national competitions, and 26.1% participated in the international competitions. Whereas the menstrual disorder was seen in 14.5% of the athletes in normal time, during the intensive exercise this ratio was increased to 20.7%. It was determined that during the competition 11.6% of the athletes used drug, 36.9% had a painful menstruation, 17.4% did not have a painful menstruation, 45.6% sometimes had a painful menstruation, and 63.1% of the athletes said that their pain decreased during the competition. First 14 days after the menstruation began, 71% of the athletes said that they felt themselves well. 71% of the athletes felt worst just before the menstruation period, 62.2% of the athletes said that their performance was same during the menstruation, and 21.2% said that their performance got worse. Both in general and during the training the menstruation period of the athletes was found to be regular (p < .01). Most of the athletes said that they have a painful menstruation period, and during the competition their pain decreased. As a result of the questionnaire, during the training and competition the number of athletes that did not use drugs were higher than the athletes that used drug (p < .01). The number of athletes that felt good before and during the menstruation were significantly higher (p < .05, p < .01). Between the menstruation periods the athletes said that they felt better in the first 14 days than the second 14 days (p < .01). When the non-menses period and menses period were compared the athletes said that their performance did not change (p < .01). It has been concluded that the menarche age was high in the athletes. It has found that the physical performance was not affected by the menstrual period and the pain decreased during the training and competition.

Am Fam Physician. 1989 Feb;39(2):213-21.
Exercise-induced amenorrhea.
Olson BR.
Strenuous exercise may cause menstrual abnormalities, including amenorrhea. The hypoestrogenemia that accompanies amenorrhea has been associated with a low bone mineral content and an increased incidence of stress fractures. With the resumption of menses, which usually occurs soon after female athletes decrease the intensity of their training or increase their body weight, bone mineral content increases and the incidence of stress fractures decreases.

Med Sci Sports Exerc. 1990 Jun;22(3):275-80.
Effects of exercise training on the menstrual cycle: existence and mechanisms.
Loucks AB.
This review evaluates the status of the evidence that exercise training affects the menstrual cycle beginning with evidence for the existence of delayed menarche, amenorrhea, and luteal suppression in athletes. A later age of menarche and a higher prevalence of amenorrhea and luteal suppression have been observed in athletes, but there is no experimental evidence that athletic training delays menarche, and alternative sociological and statistical explanations for delayed menarche have been offered. Cross-sectional studies of amenorrheic athletes have revealed abnormal reproductive hormone patterns, suggesting that the GnRH pulse generator in the hypothalamus is failing to initiate normal hypothalamic-pituitary-ovarian function. Longitudinal data show that the abrupt initiation of a high volume of aerobic training can disrupt the menstrual cycle in at least some women, but these women may be more susceptible to reproductive disruption than others, and some aspect of athletic training other than exercise (such as caloric deficiency) may be responsible for the observed disruption. Luteal suppression may be an intermediate condition between menstrual regularity and amenorrhea in athletes, or it may be the endpoint of a successful acclimation to exercise training. A potential endocrine mechanism of menstrual disruption in athletes involving the hypothalamic-pituitary-adrenal axis is discussed. Finally, promising future directions for research on this topic are described.

Appetite. 2010 Dec;55(3):379-87. Epub 2010 Aug 13.
Are female athletes at increased risk for disordered eating and its complications?
Coelho GM, Soares Ede A, Ribeiro BG.
The purpose of the study was to make a systematic review and describe and confront recent studies that compare the presence of disordered eating and its complications in young female athletes and controls subjects – PubMed, Scielo, Medline, ScienceDirect, WILEY InterScience, Lilacs and Cochrane were the databases used for this review. Out of 169 studies 22 were selected and 11,000 women from 68 sports were studied. The short version of the EAT was the most common instrument used to track disordered eating. Results showed that 55% found no significant difference in the percentage of disordered eating between athletes and controls. Also a higher percentage of studies reported higher frequency of menstrual dysfunction in athletes than controls and finally 50% of the studies found incidence of low bone mass in controls. Not all the studies that investigated all the conditions in the triad, but the authors concluded that it seemed that athletes were in more severe stage of this disorder. Due to the heterogeneity of the studies, a definitive conclusion about the groups and at highest risk for disordered eating and its complications remains to be elucidated.

Bone. 2009 Oct;45(4):760-7. Epub 2009 Jun 30.
History of amenorrhoea compromises some of the exercise-induced benefits in cortical and trabecular bone in the peripheral and axial skeleton: a study in retired elite gymnasts.
Ducher G, Eser P, Hill B, Bass S.
BACKGROUND:
Female gymnasts frequently present with overt signs of hypoestrogenism, such as late menarche or menstrual dysfunction. The objective was to investigate the impact of history of amenorrhoea on the exercise-induced skeletal benefits in bone geometry and volumetric density in retired elite gymnasts.
SUBJECTS AND METHODS:
24 retired artistic gymnasts, aged 17-36 years, who had been training for at least 15 h/week at the peak of their career and had been retired for 3-18 years were recruited. They had not been engaged in more than 2 h/week of regular physical activity since retirement. Former gymnasts who reported history of amenorrhoea (‘AME’, n=12: either primary or secondary amenorrhoea) were compared with former gymnasts (‘NO-AME’, n=12) and controls (‘C’, n=26) who did not report history of amenorrhoea. Bone mineral content (BMC), total bone area (ToA) and total volumetric density (ToD) were measured by pQCT at the radius and tibia (4% and 66%). Trabecular volumetric density (TrD) and bone strength index (BSI) were measured at the 4% sites. Cortical area (CoA), cortical thickness (CoTh), medullary area (MedA), cortical volumetric density (CoD), stress-strain index (SSI) and muscle and fat area were measured at the 66% sites. Spinal BMC, areal BMD and bone mineral apparent density (BMAD) were measured by DXA.
RESULTS:
Menarcheal age was delayed in AME when compared to NO-AME (16.4+/-0.5 years vs. 13.3+/-0.4 years, p<0.001). No differences were detected between AME and C for height-adjusted spinal BMC, aBMD and BMAD, TrD and BSI at the distal radius and tibia, CoA at the proximal radius, whereas these parameters were greater in NO-AME than C (p<0.05-0.005). AME had lower TrD and BSI at the distal radius, and lower spinal BMAD than NO-AME (p<0.05) but they had greater ToA at the distal radius (p<0.05).
CONCLUSION:
Greater spinal BMC, aBMD and BMAD as well as trabecular volumetric density and bone strength in the peripheral skeleton were found in former gymnasts without a history of menstrual dysfunction but not in those who reported either primary or secondary amenorrhoea. History of amenorrhoea may have compromised some of the skeletal benefits associated with high-impact gymnastics training.

J Clin Endocrinol Metab. 1980 Nov;51(5):1150-7.
The effects of exercise on pubertal progression and reproductive function in girls.
Warren MP.
To determine whether a significant energy drain during adolescence had a significant effect on puberty and normal reproductive function, 15 ballet dancers, aged 13–15 yr, who maintained a high level of physical activity from early adolescence were followed for 4.0 yr. Menarche was remarkably delayed in this group, occurring at a mean of 15.4 yr, significantly different (P < 0.01) from normal controls (12.5 yr) and normal music students (12.6 yr). In 2 dancers aged 18 yr, primary amenorrhea has persisted. While premenarchial, all of the dancers had varying breast development (Tanner stages 2–4) and low to low normal gonadotropin levels, normal PRL and T4 levels, and normal skull x-rays. The dancers’ mean body weight and calculated body fat were significantly less than in controls (P < 0.05). The progression of sexual development and the onset of menarche correlated in 10 or 15 subjects with a decrease in exercise and/or injury causing forced rest of at least 2-month duration. During this interval, weight gain was minimal or absent, with no significant change in body composition. A significant dichotomy in the order of pubertal development was also noted; while breast development and menarche were delayed, pubic hair development was not affected. Reversion to the amenorrheic state occurred in 11 of 13 patients with a return to exercise without a change in weight. In conclusion, energy drain may have an important modulatory effect on the hypothalamic pituitary set point at puberty and, in combination with low body weight, may prolong the prepubertal state and induce amenorrhea.

Clin Obstet Gynecol. 1985 Sep;28(3):573-9.
Body weight and the initiation of puberty.
Baker ER.
The onset and progression through the various stages of puberty are influenced by a number of factors (Fig. 2). In both animals and humans, the age of puberty appears to be related more to body weight than to chronologic age. Undernutrition and low body fat, or an altered ratio of lean mass to body fat, seem to delay the adolescent spurt and to retard the onset of menarche. According to Frisch, a minimum level of fatness (17% of body weight) is associated with menarche; however, a heavier minimum weight for height, representing an increased amount of body fat (22%), appears necessary for the onset and maintenance of regular menstrual cycles in girls over 16 years of age. This critical amount of body fat implies that a particular body composition, in addition to other environmental and psychosocial factors, is important in triggering and maintaining the pubertal process.
PIP:
Biological factors which influence the progression through female puberty stages are delineated, and an increase in the proportion of the body’s fat content is identified as a critical prerequisite for the onset and maintenance of regular ovulatory cycles. Excessive exercise or malnutrition may interfere with the normal increase in the proportion of body fat and retard the onset of menarche. Pubic hair growth and breast development begins in most American females between the ages of 8-13. Menarche follows 4.2 years later for 50% of the females, but of others, the time period ranges from 18 months to 6 mor years. Both males and females experience hormonal changes before the 1st physical signs of puberty are manifested. As sex hormones increase, changes in the body’s proportion of lean, fat, and skeletal mass occur. For females an increase in body fat begins at 7 years and continues through ages 16-18 years. Studies indicate that the body’s fat content must account for 17% of the body’s weight before menarche can occur and that, at age 18 years, the fat content must be at least 22% for the maintenance of regular menstrual cycles. Apparently, hypothalamic sensitivity to estrogens is decreased when the critical ratio of lean mass to body fat is reached, and changes in the hypothalamic and pituitary hormones promote pubertal progression and the establishment of reproductive functions. Poor nutrition alters the ratio of lean mass to body fat and delays the onset of menarche. In the US, the age at menarche decreased by 3 years since 1840 due to improvements in the population’s nutritional status. Underweight females generally experience menarche at later ages than normal weight females. In contrast overweight females often experience menarche earlier than the average weight female. Athletic females and ballet dancers frequently experience late menarche, and these delays may be due to the disruption in fat accumulation which results from excessive exercise. Physically, inactive adolescents, on the other hand, tend to experience menarche at an earlier age than normally active females. In conclusion, the body’s fat content along with a variety of environmental and psychosocial factors are responsible for the development and maintenance of female reproductive functions.

J Sports Med Phys Fitness. 1996 Mar;36(1):49-53.
Gymnasts, distance runners, anorexics body composition and menstrual status.
Bale P, Doust J, Dawson D.
Ten top class female distance runners, ten female anorexics and twenty female gymnasts of a similar age were compared for height, mass, %fat, fat mass, lean body mass, age of menarche and incidence of amenorrhoea. The mean age of the distance runners, anorexics, and gymnasts was 13.6 years, 14.7 years, and 13.3 years respectively. In comparison to normal data on females of a similar age they were shorter, lighter, had lower fat masses, and %fat, and the gymnasts and anorexics had lower lean body masses. However, the gymnasts and runners had higher lean body masses compared with the anorexic group. There were no significant differences in body composition by hydrostatic weighing but of these three groups the anorexics tended to have the highest total skinfold, %fat and fat masses. Only 20% of the gymnasts, 40% of the runners and 70% of the anorexics had started menstruating compared with 95% of girls of a similar age. Of the girls in our study who had started menstruating one gymnast, (25% of sub-group) two runners (50% of sub-group) and seven anorexics (100% of sub-group) had developed secondary amenorrhoea. The low body masses, low fat masses, delayed menarche and secondary amenorrhoea in athletes are discussed in relation to low caloric intake, stress, hormone levels, high training loads and genetic factors. Our data demonstrating no significant differences in body composition variables between the three groups of young girls, support the main contention that this type of physique may arise through different mechanisms leading to a common outcome, but without a proven causal link between anorexia and athletic performance.

Am Fam Physician. 1996 Mar;53(4):1185-94.
Evaluation of amenorrhea.
Kiningham RB, Apgar BS, Schwenk TL.
Pregnancy is the most common cause of amenorrhea and must be ruled out before proceeding with diagnostic evaluation. A careful history and physical examination may reveal evidence of androgen excess, estrogen deficiency or other endocrinopathies. Serum prolactin and thyroid-stimulating hormone (TSH) levels should be checked in all women who are not pregnant. Galactorrhea by history or on examination and/or an elevated prolactin level should be investigated with an imaging study to rule out a pituitary adenoma. If serum prolactin and TSH levels are normal, a progesterone challenge test should be performed to determine outflow tract patency and estrogen status. In women with hypoestrogenic amenorrhea, indicated by a negative challenge test and a competent outflow tract, serum gonadotropin, follicle-stimulating hormone and luteinizing hormone levels may be measured to determine whether amenorrhea represents ovarian failure or pituitary or hypothalamic dysfunction. Hypothalamic amenorrhea is common in women with a history of weight loss, stress or vigorous exercise. Amenorrheic women with adequate estrogen levels should receive cyclic progesterone. Hormonal therapy and calcium supplementation in hypoestrogenic amenorrhea.

Obstet Gynecol. 1979 Jul;54(1):47-53.
Menstrual dysfunction in distance runners.
Dale E, Gerlach DH, Wilhite AL.
The problem of menstrual dysfunction in women who engage in endurance training for participation in distance running events has been studied. Through survey, selected aspects of the personal, training, menstrual, and contraceptive histories of 168 women who were defined as runners, joggers, or controls were evaluated. In addition, defined subsets of the study subjects were evaluated for serum levels of pituitary and ovarian hormones and determination of percentage body fat. The data show significant differences among the 3 groups. It is concluded that menstrual dysfunction in distance runners is a real phenomenon. Presumably this is related to decreased percentage of body fat and/or minimal ovarian function secondary to diminished hypothalamic or pituitary hormone secretion.

Ugeskr Laeger. 1994 Nov 28;156(48):7219-23.
[Bone metabolism in female runners. Menstruation disorders are frequent among long-distance runners, but the bone mass is not influenced, with the exception of runners with amenorrhea].
[Article in Danish]
Hetland ML, Haarbo J, Christiansen C, Larsen T.
The purpose of the study was to investigate the prevalence of exercise-related menstrual and sex hormonal disturbances and the effect of exercise on bone mass and metabolism in female runners at various training levels. Two hundred and five premenopausal women (running 0-140 km a week) were recruited from a large population of female runners, who had responded to a questionnaire regarding exercise habits. Maximum oxygen uptake was determined by treadmill testing. Gynaecological status was assessed on entries in a menstrual calendar and by transvaginal ultrasonography; and sex hormonal status was measured three times with 10-day intervals. Bone mass was measured in the lumbar spine, proximal femurs and total body by dual energy x-ray absorptiometry, and in the forearm by single photon absorptiometry. Bone turnover was assessed by plasma osteocalcin, serum alkaline phosphatase, and urinary calcium and hydroxyproline. The results showed that sex hormonal disturbances were significantly related to training intensity. Compared with the normally active women, the baseline levels and fluctuations of oestradiol and progesterone in the elite runners were reduced by up to 25-44%, (0.01 < p < 0.05). The prevalence of amenorrhoea increased from 1% in the normally active to 11% in the elite runners. No statistically significant relation was found between running activity and bone mass or bone turnover. However, the group of amenorrhoeic runners had a 10% reduction in lumbar bone mass as compared to the normally menstruating runners (p < 0.05), but the bone turnover was similar.(ABSTRACT TRUNCATED AT 250 WORDS)

Am J Med. 1993 Jul;95(1):53-60.
Running induces menstrual disturbances but bone mass is unaffected, except in amenorrheic women.
Hetland ML, Haarbo J, Christiansen C, Larsen T.
PURPOSE:
To investigate the prevalence of exercise-related menstrual and sex hormonal disturbances and the effect of exercise on bone mass and metabolism in female runners at various training levels.
SUBJECTS AND METHODS:
Two hundred five premenopausal women (running 0 to 140 km a week) were recruited from a large population of female runners who had responded to a questionnaire regarding exercise habits. Maximum oxygen uptake was determined by treadmill testing. Gynecologic status was assessed on entries in a menstrual calendar and by transvaginal ultrasonography; sex hormonal status was measured three times with 10-day intervals. Bone mass was measured in the lumbar spine, proximal femurs, and total body by dual-energy x-ray absorptiometry, and in the forearm by single-photon absorptiometry. Bone turnover was assessed by measurement of plasma osteocalcin, serum alkaline phosphatase, and urinary calcium and hydroxyproline.
RESULTS:
Sex hormonal disturbances were significantly related to training intensity. Compared with the normally active women, the baseline levels and fluctuations of estradiol and progesterone in the elite runners were reduced by up to 25% to 44% (0.01 < p < 0.05). The prevalence of amenorrhea increased from 1% in the normally active subjects to 11% in the elite runners. No statistically significant relation was found between running activity and bone mineral measurements or bone turnover. However, the group of amenorrheic runners had a 10% reduction in lumbar bone density as compared with the normally menstruating runners (p < 0.05), but the bone turnover was similar. CONCLUSION: In the large majority of the female runners, no skeletal affection was found despite significant sex hormonal and menstrual disturbances. Only the runners with amenorrhea might be at increased risk of osteoporosis.

Fertil Steril. 1981 Dec;36(6):691-6.
Menstrual dysfunction and hormonal status in athletic women: a review.
Baker ER.
Since women have become more involved in physical fitness and competitive endurance sports, the incidence of menstrual dysfunction has increased. Long-distance running and other sports may lead to alterations in gonadotropins, androgens, estrogens, progesterone, or prolactin, which in some women may directly or indirectly result in amenorrhea or infertility. The effects of running and strenuous exercise on the menstrual cycle and reproductive hormones remain controversial. Reported incidences of menstrual dysfunction vary widely, and many factors have been implicated in the onset of this problem. Exercise associated menstrual dysfunction seems to occur more frequently in nulliparous athletes, in athletes with delayed menarche, and in athletes with low body fat. It is important to realize that disruption of the menstrual cycle, ranging from mild changes in flow to amenorrhea, is a relatively common problem for the female athlete engaged in strenuous endurance sports. Yet no evidence exists at present to indicate conclusively that this menstrual dysfunction is harmful to the female athlete’s reproductive system.

South Med J. 1983 May;76(5):619-24.
Athletic activity and menstruation.
Diddle AW.
Menstrual dysfunction characterized by delayed menarche, irregular menses, or secondary amenorrhea often affects women who compete in athletics over a prolonged time. Loss of body fat and emotional stress are important predisposing factors. Under these circumstances, hypoestrogenism, an altered ratio of follicle-stimulating hormone to luteinizing hormone, and elevation of serum testosterone, prolactin, catecholamines, and opioids are fairly common. There is controversy over whether the working capacity and performance of the average woman varies appreciably during various phases of the menstrual cycle. Apparently, those who suffer from premenstrual tension do have a diminution in working capacity. Generally, the causes of menstrual dysfunction are the same for athletes and nonathletes, and there are currently no data to forbid athletes’ training at any time during the menses. A gynecologic examination should be done before menstrual dysfunction is considered to be due to physical exercise. If this assumption is substantiated, and if amenorrhea persists for one year or more, a periodic estrogen-progesterone regimen should be offered to minimize vascular problems, osteoporosis, and stress fractures, and to protect the endometrium and ovarian function.

Br J Sports Med. 2005 Mar;39(3):141-7.
Participation in leanness sports but not training volume is associated with menstrual dysfunction: a national survey of 1276 elite athletes and controls.
Torstveit MK, Sundgot-Borgen J.
OBJECTIVE:
To examine the prevalence of menstrual dysfunction in the total population of Norwegian elite female athletes and national representative controls in the same age group.
METHODS:
A detailed questionnaire that included questions on training and/or physical activity patterns, menstrual, dietary, and weight history, oral contraceptive use, and eating disorder inventory subtests was administered to all elite female athletes representing the country at the junior or senior level (aged 13-39 years, n = 938) and national representative controls in the same age group (n = 900). After exclusion, a total of 669 athletes (88.3%) and 607 controls (70.2%) completed the questionnaire satisfactorily.
RESULTS:
Age at menarche was significantly (p<0.001) later in athletes (13.4 (1.4) years) than in controls (13.0 (1.3) years), and differed among sport groups. A higher percentage of athletes (7.3%) than controls (2.0%) reported a history of primary amenorrhoea (p<0.001). A similar percentage of athletes (16.5%) and controls (15.2%) reported present menstrual dysfunction, but a higher percentage of athletes competing in leanness sports reported present menstrual dysfunction (24.8%) than athletes competing in non-leanness sports (13.1%) (p<0.01) and controls (p<0.05).
CONCLUSIONS:
These novel data include virtually all eligible elite athletes, and thus substantially extend previous studies. Age at menarche occurred later and the prevalence of primary amenorrhoea was higher in elite athletes than in controls. A higher percentage of athletes competing in sports that emphasise thinness and/or a specific weight reported present menstrual dysfunction than athletes competing in sports focusing less on such factors and controls. On the basis of a comparison with a previous study, the prevalence of menstrual dysfunction was lower in 2003 than in 1993.

Gynecol Obstet Invest. 2000;49(1):41-6.
Women endurance runners with menstrual dysfunction have prolonged interruption of training due to injury.
Beckvid Henriksson G, Schnell C, Lindén Hirschberg A.
Strenuous exercise by women is associated with menstrual dysfunction, eating disorders and osteoporosis. Intensive training may also increase the susceptibility to infections. In this study, we investigated whether menstrual dysfunction was related to musculoskeletal injuries and/or upper respiratory tract infections in women middle/long-distance runners. A questionnaire was mailed to 127 Swedish female runners of whom 75% answered. This retrospective study showed a higher frequency of menstrual disorders (25%) in runners than in the general population. Furthermore, almost half of the athletes (46%) were classified as at risk of developing eating disorders. Women athletes with menstrual dysfunction were found to have had a longer interruption of training due to musculoskeletal injuries than those with regular cycles (34.1 +/- 3.0 vs. 9.0 +/- 9. 4 days, p < 0.05). However, no relation was found between susceptibility to infections and menstrual status.

Am J Obstet Gynecol. 1982 Aug 15;143(8):862-9.
The effect of marathon training upon menstrual function.
Shangold MM, Levine HS.
Detailed questionnaires were distributed to the 1,841 women who entered the 1979 New York City Marathon; the questions pertained to obstetric, gynecologic, and athletic histories, as well as height and weight. The incidence of oligomenorrhea/amenorrhea among the 394 respondents was 24% during training and 19% prior to training. The incidence of infertility among respondents was 10%. Of those women who had had regular menses prior to training, 93% continued to have regular menses during training. Amenorrheic women were significantly lighter (P less than 0.005) than regularly menstruating women and had significantly lower weight/height ratios (P less than. 0.0005). The best predictor of a women’s menstrual pattern during training was her pretraining menstrual pattern. Thinness was associated with amenorrhea, regardless of training.

Am J Physiol. 1994 Mar;266(3 Pt 2):R817-23.
Induction of low-T3 syndrome in exercising women occurs at a threshold of energy availability.
Loucks AB, Heath EM.
To investigate the relationship between energy availability (dietary energy intake minus energy expended during exercise) and thyroid metabolism, we studied 27 untrained, regularly menstruating women who performed approximately 30 kcal.kg lean body mass (LBM)-1.day-1 of supervised ergometer exercise at 70% of aerobic capacity for 4 days in the early follicular phase. A clinical dietary product was used to set energy availability in four groups (10.8, 19.0, 25.0, 40.4 kcal.kg LBM-1.day-1). For 9 days beginning 3 days before treatments, blood was sampled once daily at 8 A.M. Initially, thyroxine (T4) and free T4 (fT4), 3,5,3′-triiodothyronine (T3) and free T3 (fT3), and reverse T3 (rT3) were in the normal range for all subjects. Repeated-measures one-way analysis of variance followed by one-sided, two-sample post hoc Fischer’s least significant difference tests of changes by treatment day 4 revealed that reductions in T3 (16%, P < 0.00001) and fT3 (9%, P < 0.01) occurred abruptly between 19.0 and 25.0 kcal.kg LBM-1.day-1 and that increases in fT4 (11%, P < 0.05) and rT3 (22%, P < 0.01) occurred abruptly between 10.8 and 19.0 kcal.kg LBM-1.day-1. Changes in T4 could not be distinguished. If energy deficiency suppresses reproductive as well as thyroid function, athletic amenorrhea might be prevented or reversed by increasing energy availability through dietary reform to 25 kcal.kg LBM-1.day-1, without moderating the exercise regimen.

Am J Physiol. 1993 May;264(5 Pt 2):R924-30.
Induction and prevention of low-T3 syndrome in exercising women.
Loucks AB, Callister R.
To investigate the influence of exercise on thyroid metabolism, 46 healthy young regularly menstruating sedentary women were randomly assigned to a 3 x 2 experimental design of aerobic exercise and energy availability treatments. Energy availability was defined as dietary energy intake minus energy expenditure during exercise. After 4 days of treatments, low energy availability (8 vs. 30 kcal.kg body wt-1.day-1) had reduced 3,5,3′-triiodothyronine (T3) by 15% and free T3 (fT3) by 18% and had increased thyroxine (T4) by 7% and reverse T3 (rT3) by 24% (all P < 0.01), whereas free T4 (fT4) was unchanged (P = 0.08). Exercise quantity (0 vs. 1,300 kcal/day) and intensity (40 vs. 70% of aerobic capacity) did not affect any thyroid hormone (all P > 0.10). That is, low-T3 syndrome was induced by the energy cost of exercise and was prevented in exercising women by increasing dietary energy intake. Selective observation of low-T3 syndrome in amenorrheic and not in regularly menstruating athletes suggests that exercise may compromise the availability of energy for reproductive function in humans. If so, athletic amenorrhea might be prevented or reversed through dietary reform without reducing exercise quantity or intensity.

=======================
Effect of luteal deficiency.

Acta Obstet Gynecol Scand. 1971;50(1):61-2.
Luteal insufficiency and pelvic adhesions.
Johansson ED, Persson BH, Gemzell C.
A young woman with a history of a septic abortion and left oophorectomy for a dermoid cyst was investigated before and after laparotomy with regard to the function of the corpus luteum. At laparotomy the remaining right ovary was surrounded and fixed by thick adhesions around the ovary. The adhesions were removed. During two regular menstrual cycles before operation, low plasma levels of progesterone were found during the luteal phase. After the removal of the adhesions normal plasma levels of progesterone were found. The urinary excretion of oestrogens also improved. Severe pelvic adhesions might be one cause of insufficient luteal function.

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Endotoxin-lipoprotein Hypothesis

Also see:
Ray Peat, PhD on the Benefits of the Raw Carrot
Endotoxin: Poisoning from the Inside Out
Protective Bamboo Shoots
The effect of raw carrot on serum lipids and colon function
Protection from Endotoxin
The Truth about Low Cholesterol

“The rate of cholesterol production, and the amount in circulation, tend to be inversely related to systemic inflammation. All of the types of lipoprotein absorb, bind, and help to eliminate endotoxin, for example.” -Ray Peat, PhD

“Cholesterol has a long history as a protectant against many toxins; I think this relates to the fact that people with very low cholesterol have such a high incidence of endotoxin-related symptoms.” -Ray Peat, PhD

Lancet. 2000 Sep 9;356(9233):930-3.
The endotoxin-lipoprotein hypothesis.
Rauchhaus M, Coats AJ, Anker SD.
The advent of 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors (statins) has revolutionised the treatment of hypercholesterolaemia. Statin treatment, by lowering the atherogenic lipoprotein profile, reduces morbidity and mortality in patients with cardiovascular disease. Treatment with simvastatin causes a reduction of events of new-onset heart failure, but this may be attributable to properties other than its lipid-lowering effects. There is some evidence that lower serum cholesterol concentrations (as a surrogate for the totality of lipoproteins) relate to impaired survival in patients with chronic heart failure (CHF). Inflammation is a feature in patients with CHF and increased lipopolysaccharide may contribute substantially. We postulate that higher concentrations of total cholesterol are beneficial in these patients. This is potentially attributable to the property of lipoproteins to bind lipopolysaccharide, thereby preventing its detrimental effects. We hypothesise there is an optimum lipoprotein concentration below which lipid reduction would, on balance, be detrimental. We also propose that, in patients with CHF, a non-lipid-lowering statin (with ancillary properties such as immune modulatory and anti-inflammatory actions) could be as effective or even more beneficial than a lipid-lowering statin.

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Oral Contraceptives, Estrogen, and Clotting

Also see:
High Estrogen and Heart Disease in Men
Aldosterone and Thrombosis

In the 1970s, after reading Szent-Gyorgyi’s description of the antagonistic effect of progesterone and estrogen on the heart, I reviewed the studies that showed that progesterone protects against estrogen’s clotting effect. I experimented with progesterone, showing that it increases the muscle tone in the walls of veins, which is very closely related to the effects Szent-Gyorgyi described in the heart. And progesterone opposes estrogen’s ability to increase the amount of free fatty acids circulating in the blood. -Ray Peat, PhD

Br Med J. 1970 April 25; 2(5703): 203–209.
Thromboembolic Disease and the Steroidal Content of Oral Contraceptives. A Report to the Committee on Safety of Drugs
W. H. W. Inman, M. P. Vessey, Barbro Westerholm, and A. Engelund
Reports of thromboembolism following the use of oral contraceptives received by drug safety committees in the United Kingdom, Sweden, and Denmark have been analysed to investigate possible differences in the risks associated with the various preparations. For this purpose the numbers of reports of thromboembolism attributed to each product were compared with the distribution that would have been expected from market research estimates of sales, assuming that all products carried the same risk.

A positive correlation was found between the dose of oestrogen and the risk of pulmonary embolism, deep vein thrombosis, cerebral thrombosis, and coronary thrombosis in the United Kingdom. A similar association was found for venous thrombosis and pulmonary embolism in Sweden and Denmark.
No significant differences could be detected between sequential and combined preparations containing the same doses of oestrogen, nor between the two oestrogens, ethinyloestradiol and mestranol.

Certain discrepancies in the data suggest that the dose of oestrogen may not be the only factor related to the risk of thromboembolism; thus there was a significant deficit of reports associated with the combination of mestranol 100 μg. with norethynodrel 2·5 mg. and a significant excess of reports associated with the combination of ethinyloestradiol 50 μg. with megestrol acetate 4 mg. An excess of reports also occurred with other combined preparations containing megestrol acetate.

The data obtained in earlier epidemiological studies were re-examined and, though no trend was obvious in any one of them, the combined results showed an excess of cases of thromboembolism at the highest dose of oestrogen.

Lancet. 1976 Mar 6;1(7958):509-11.
Oral contraceptives, antithrombin- III activity, and postoperative deep-vein thrombosis.
Sagar S, Stamatakis JD, Thomas DP, Kakkar VV.
Deep-vein thrombosis (D.V.T.) was detected by the fibrinogen-uptake test in six out of a total of thirty-one young women undergoing emergency abdominal surgery who gave a history of recent oral contraceptive intake. In contrast, no D.V.T. developed in nineteen similar patients who were not on oral contraceptives (P less than 0.01). Plasma-antithrombin-III activity was significantly lower preoperatively in patients taking oral contraceptives; postoperative D.V.T. subsequently developed in three out of five patients with preoperative antithrombin-III activity below 50%. In seventy-eight dental patients undergoing molar extraction, antithrombin-III activity was measured before, during, and after operation. Activity fell in all patients during operation, but the fall was significantly greater in women taking oral contraceptives (P less than 0.01). The intra-operative fall in antithrombin-III activity was prevented by a small preoperative dose of subcutaneous heparin.

Am J Obstet Gynecol. 1975 Jul 15;122(6):688-92.
Conjugated estrogens and hypercoagulability.
von Kaulla E, Droegemueller W, von Kaulla KN.
A group of 11 menopausal women receiving 1.25 mg. of conjugated estrogens daily had coagulation tests to determine the development of hypercoagulability after taking 5 and 21 tablets. There was no essential change in thrombin generation or fibrinolytic activity as measured by euglobin lysis time. There was a shift toward hypercoagulability in all three parameters of the thrombelastograms. The decrease of the antithrombin III activity was not as pronounced following the administration of conjugated estrogens as had been the change associated with oral contraceptives. Fibrin monomers were observed in some women during the first week of Premarin therapy.

Arch Pathol. 1970 Jan;89(1):1-8.
Vascular lesions in women taking oral contraceptives.
Irey NS, Manion WC, Taylor HB.
Distinctive vascular lesions in association with thrombosis were found in arteries and veins in 20 relatively young women receiving oral contraceptives. These lesions were characterized by structural and histochemical changes in the intima and media. Occlusive thrombi were associated with relatively small, organized bases, the age of the latter measured in days to weeks. Nonocclusive and possibly earlier lesions were dominated by endothelial proliferation with minimal thrombus formation. It is postulated that this endothelial and intimal hyperplasia may be related to the steroids received and that it parallels similarly induced hyperplasias that have been found in cervical gland epithelium, in leiomyomas, and in a variety of mesenchymal derivatives under experimental conditions. Further control and experimental studies are required to clarify the possible relationship between these vascular lesions and oral contraceptives.

Br Med J. 1973 December 1; 4(5891): 507–512.
Cryptogenic Cerebral Embolism in Women Taking Oral Contraceptives
Karin Enzell and Gunnar Lindemalm
Fourteen women taking oral contraceptives were admitted during a five-year period because of acute cerebrovascular lesions. A diagnosis of major cerebral embolism was established in four of them. No source of embolism was found, and thorough investigation failed to reveal any predisposing illness. Cerebral embolism was a probable diagnosis in several of the remaining 10 patients. A comparison was made with the strokes occurring in women not taking contraceptive pills in corresponding age groups.

Lancet. 1973 Jun 23;1(7817):1399-404.
Oral contraceptives and venous thromboembolic disease, surgically confirmed gallbladder disease, and breast tumours. Report from the Boston Collaborative Drug Surveillance Programme.
[No authors listed]
A large survey of 24 hospitals was conducted to identify associations between commonly used drugs and various diseases. The results of 3 such studies–on venous thromboembolism, gall bladder disease, and breast tumors–are summarized in this article. Trained nurses in various hospital wards interviewed admissions, asking questions designed to determine smoking behavior, coffee and tea drinking, drug use, marital status, and parity and menopausal status, where appropriate. This report specifically centers on associations between oral contraceptive use and development of the 3 conditions under study. Women reported on in this portion of the study were aged 20-44 years. Compared with nonusers, the estimate of relative risk for thromboembolism in users was 11, and the estimated attack rate attributable to oral contraceptives was 60/100,000 users/year. For gall bladder disease (surgically confirmed) the corresponding relative risk estimate was 2.0, and the estimated annual attack rate was 79/100,000. The frequency of gall bladder disease in women under 35 years was significantly higher in oral contraceptive users of 6-12 months duration, compared with women who had taken the pills for longer periods. Breast cancer studies showed no evidence of a higher risk in oral contraceptive users relative to nonusers. In fact, a negative association between oral contraceptive use and breast tumors was found, and this was more pronounced in women with fibroadenoma of the breast. Most of the women surveyed for this report took low-dose estrogen formulations, but the role of dose to the above findings was not investigated.

The finding of a positive correlation between the dose of oestrogen and the risk of coronary thrombosis is of special interest since previous studies have failed to provide clear evidence of a relationship between oral contraceptives and this condition.

Am J Obstet Gynecol. 1987 Oct;157(4 Pt 2):1042-8.
Coagulation effects of oral contraception.
Bonnar J.
In Europe and North America, estrogen/progestogen oral contraception has been associated with an increase in venous thromboembolism, myocardial infarction, and stroke. These hazards are found mainly in smokers and in women over the age of 35. Venous thromboembolism appears to correlate with the estrogen dosage, and the arterial complications with both the estrogen and progestogen components. Blood coagulation and vascular thrombosis are intimately related. Estrogen/progestogen oral contraception affects blood clotting by increasing plasma fibrinogen and the activity of coagulation factors, especially factors VII and X; antithrombin III, the inhibitor of coagulation, is usually decreased. Platelet activity is also enhanced with acceleration of aggregation. These changes create a state of hypercoagulability that, to a large extent, appears to be counterbalanced by increased fibrinolytic activity. Studies of the oral contraceptives in current use show that the coagulation effects depend on the dosage of estrogen and the type of progestogen used in combination. Current research is aimed at finding the estrogen/progestogen formulations that induce the least changes in the coagulation system and other physiologic processes. In this respect, the new low-dose formulations are a major step forward and should reduce the risk of vascular thrombotic complications.

Lancet. 1980 May 24;1(8178):1097-101.
Oral contraceptives and thromboembolic disease: effects of lowering oestrogen content.
Böttiger LE, Boman G, Eklund G, Westerholm B.
The introduction of low-oestrogen oral contraceptives in Sweden and the concomitant disappearance of high-dose preparations did not result in a lowering of the mortality of fertile women from thromboembolic disease. Morbidity due to thromboembolism seems to have fallen, and the number of thromboembolic incidents reported to the Swedish Adverse Drug Reaction Committee decreased dramatically. The decrease was due exclusively to a reduction in venous thromboembolic disease: the frequency of arterial complications (cerebral and coronary) remained constant.

Estrogen has many pro-clotting effects, and one of them is a decreased activity of vascular plasminogen activator. K. E. Miller and S. V. Pizzo, “Venous and arterial thromboembolic disease in women using oral contraceptives,” Am. J. Obst. Gyn. 144, 824, 1982. -Ray Peat, PhD

Am J Obstet Gynecol. 1982 Dec 1;144(7):824-7.
Venous and arterial thromboembolic disease in women using oral contraceptives.
Miller KE, Pizzo SV.
Vascular plasminogen activator was measured by means of a new chromogenic assay in 24 women who had suffered from oral contraceptive-associated thrombotic disease and was compared to that in a control group of 78 premenopausal women. Vascular plasminogen activator levels were significantly reduced in the subjects who had venous thrombosis but not in the five women who had arterial thrombosis (0.04 +/- 0.03 versus 0.38 +/- 0.31, respectively) when compared to the levels in the control group (0.19 +/- 0.20). Since vascular activator levels distribute in a non-Gaussian manner, cases and controls were also stratified into deciles. Seventeen subjects who had suffered from venous thrombosis were stratified in the lowest three deciles, and two subjects, in the fourth and fifth deciles. Subjects who had suffered from arterial thrombosis were in the fourth or higher deciles. The conclusion is that, although there is a correlation between low levels of vascular plasminogen activator and venous thrombosis, no such correlation exists for arterial thrombosis.

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Protective Bamboo Shoots

Also see:
Ray Peat, PhD on the Benefits of the Raw Carrot
The effect of raw carrot on serum lipids and colon function
Endotoxin: Poisoning from the Inside Out
Endotoxin-lipoprotein Hypothesis
Protection from Endotoxin
Bowel Toxins Accelerate Aging

“The food industry is promoting the use of various gums and starches, which are convenient thickeners and stabilizers for increasing self-life, with the argument that the butyric acid produced when they are fermented by intestinal bacteria is protective. However, intestinal fermentation increases systemic and brain serotonin, and the short-chain fatty acids can produce a variety of inflammatory and cytotoxic effect. Considering the longevity and stress-resistance of germ-free animals, choosing foods (such as raw carrots or cooked bamboo shoots or cooked mushrooms) which accelerate peristalsis and speed transit through the bowel, which suppressing bacterial growth, seems like a convenient approach to increasing longevity.” -Ray Peat, PhD

Comprehensive Reviews in Food Science and Food Safety Volume 10, Issue 3, pages 153–168, May 2011
Nutritional Properties of Bamboo Shoots: Potential and Prospects for Utilization as a Health Food
Nirmala Chongtham, Madho Singh Bisht, Sheena Haorongbam
Bamboo is intricately associated with humans from times immemorial. Popularly known for their industrial uses, a lesser known fact of bamboos is the usage of its young shoots as a food that can be consumed fresh, fermented, or canned. The juvenile shoots are not only delicious but are rich in nutrient components, mainly proteins, carbohydrates, minerals, and fiber and are low in fat and sugars. In addition, they contain phytosterols and a high amount of fiber that can be labeled as nutraceuticals or natural medicines that are attracting the attention of health advocates and scientists alike. The shoots are free from residual toxicity and grow without the application of fertilizers. Modern research has revealed that bamboo shoots have a number of health benefits: improving appetite and digestion, weight loss, and curing cardiovascular diseases and cancer. The shoots are reported to have anticancer, antibacterial, and antiviral activity. Shoots have antioxidant capacity due to the presence of phenolic compounds. The increasing trends of health consciousness among consumers have stimulated the field of functional foods and bamboo shoots can be one of them. Bamboo fiber is now a common ingredient in breakfast cereals, fruit juices, bakery and meat products, sauces, shredded cheeses, cookies, pastas, snacks, frozen desserts, and many other food products. This review emphasizes the health benefits of bamboo shoots and their potential for utilization as a health food.

Nutrition. 2009 Jul-Aug;25(7-8):723-8. Epub 2009 Mar 13.
Effects of bamboo shoot consumption on lipid profiles and bowel function in healthy young women.
Park EJ, Jhon DY.
OBJECTIVE:
This study evaluated the short-term effect of bamboo shoot consumption as a dietary fiber source on blood glucose, lipid profiles, hepatic function, and constipation symptoms in healthy women.
METHODS:
Eight subjects, 21- to 23-y-old women, with normal health status received a dietary fiber-free diet (control), a diet containing 25 g of cellulose, and a diet containing 360 g of bamboo shoots, with each diet segment lasting 6 d. At the end of each diet, blood biochemical parameters, such as glucose, triacylglycerols, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, glutamic pyruvic transaminase, glutamic oxaloacetic transaminase, and atherogenic index were measured and a questionnaire test for the evaluation of fecal excretion was taken. For statistical analysis, analysis of variance was performed.
RESULTS:
Serum total cholesterol, low-density lipoprotein cholesterol, and the atherogenic index were decreased with the bamboo shoot diet feeding compared with the dietary fiber-free diet. There were no differences in serum glucose levels among the tested diets. Fecal volume and bowel movement frequency in subjects fed the bamboo shoot diet were significantly increased.
CONCLUSION:
Bamboo shoots as a dietary fiber source has beneficial effects on lipid profile and bowel function.

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Calcium to Phosphorus Ratio, PTH, and Bone Health

Also see:
Calcium Paradox
Source of Dietary Calcium: Chicken Egg Shell Powder
Low CO2 in Hypothyroidism
Blood Pressure Management with Calcium & Dairy
Hypertension and Calcium Deficiency
Excess Dietary Phosphorus Lowers Vitamin D Levels
Fatty Acid Synthase (FAS), Vitamin D, and Cancer
Phosphate, activation, and aging
Parmigiano Reggiano cheese and bone health
Dairy, Calcium, and Weight Management in Adults and Children

Picture 3

Quotes by Ray Peat, PhD:
“The ratio of calcium to phosphate is very important; that’s why milk and cheese are so valuable for weight loss, or for preventing weight gain. For people who aren’t very active, low fat milk and cheese are better, because the extra fat calories aren’t needed.”

“The foods highest in phosphate, relative to calcium, are cereals, legumes, meats, and fish. Many prepared foods contain added phosphate. Foods with a higher, safe ratio of calcium to phosphate are leaves, such as kale, turnip greens, and beet greens, and many fruits, milk, and cheese.”

“Free unsaturated fatty acids turn on the stress hormones, and cortisol blocks oxidation of sugar and turns it into fatty acids and triglycerides. Keeping cortisol and stress low is the main thing. Keeping a high ratio of calcium to phosphate helps to oppose the stress metabolism.”

“Recent publication are showing that excess phosphate can increase inflammation, tissue atrophy, calcification of blood vessels, cancer, dementia, and, in general, the processes of aging.”

Br J Nutr. 2006 Sep;96(3):545-52.
High phosphorus intakes acutely and negatively affect Ca and bone metabolism in a dose-dependent manner in healthy young females.
Kemi VE, Kärkkäinen MU, Lamberg-Allardt CJ.
Ca and P are both essential nutrients for bone and are known to affect one of the most important regulators of bone metabolism, parathyroid hormone (PTH). Too ample a P intake, typical of Western diets, could be deleterious to bone through the increased PTH secretion. Few controlled dose-response studies are available on the effects of high P intake in man. We studied the short-term effects of four P doses on Ca and bone metabolism in fourteen healthy women, 20-28 years of age, who were randomized to four controlled study days; thus each study subject served as her own control. P supplement doses of 0 (placebo), 250, 750 or 1500 mg were taken, divided into three doses during the study day. The meals served were exactly the same during each study day and provided 495 mg P and 250 mg Ca. The P doses affected the serum PTH (S-PTH) in a dose-dependent manner (P=0.0005). There was a decrease in serum ionized Ca concentration only in the highest P dose (P=0.004). The marker of bone formation, bone-specific alkaline phosphatase, decreased (P=0.05) and the bone resorption marker, N-terminal telopeptide of collagen type I, increased in response to the P doses (P=0.05). This controlled dose-response study showed that P has a dose-dependent effect on S-PTH and increases PTH secretion significantly when Ca intake is low. Acutely high P intake adversely affects bone metabolism by decreasing bone formation and increasing bone resorption, as indicated by the bone metabolism markers.

Br J Nutr. 2008 Apr;99(4):832-9. Epub 2007 Oct 1.
Increased calcium intake does not completely counteract the effects of increased phosphorus intake on bone: an acute dose-response study in healthy females.
Kemi VE, Kärkkäinen MU, Karp HJ, Laitinen KA, Lamberg-Allardt CJ.
A high dietary P intake is suggested to have negative effects on bone through increased parathyroid hormone secretion, as high serum parathyroid hormone (S-PTH) concentration increases bone resorption. In many countries the P intake is 2- to 3-fold above dietary guidelines, whereas Ca intake is too low. This combination may not be optimal for bone health. In a previous controlled study, we found that dietary P dose-dependently increased S-PTH and bone resorption and decreased bone formation. The aim of the present study was to investigate the dose-response effects of Ca intake on Ca and bone metabolism with a dietary P intake higher than recommended.
Each of the twelve healthy female subjects aged 21-40 years attended three 24-h study sessions, which were randomized with regard to a Ca dose of 0 (control day), 600 or 1200 mg, and each subject served as her own control. The meals on each study day provided 1850 mg P and 480 mg Ca. S-PTH concentration decreased (P < 0.001) and serum ionized Ca concentration increased (P < 0.001) with increasing Ca doses. The bone formation marker, serum bone-specific alkaline phosphatase, did not differ significantly (P = 0.4). By contrast, the bone resorption marker, urinary N-terminal telopeptide of collagen type I, decreased significantly with both Ca doses (P = 0.008). When P intake was above current recommendations, increased Ca intake was beneficial for bone, as indicated by decreased S-PTH concentration and bone resorption. However, not even a high Ca intake could affect bone formation when P intake was excessive.

Br J Nutr. 2010 Feb;103(4):561-8. Epub 2009 Sep 28.
Low calcium:phosphorus ratio in habitual diets affects serum parathyroid hormone concentration and calcium metabolism in healthy women with adequate calcium intake.
Kemi VE, Kärkkäinen MU, Rita HJ, Laaksonen MM, Outila TA, Lamberg-Allardt CJ.
Excessive dietary P intake alone can be deleterious to bone through increased parathyroid hormone (PTH) secretion, but adverse effects on bone increase when dietary Ca intake is low. In many countries, P intake is abundant, whereas Ca intake fails to meet recommendations; an optimal dietary Ca:P ratio is therefore difficult to achieve. Our objective was to investigate how habitual dietary Ca:P ratio affects serum PTH (S-PTH) concentration and other Ca metabolism markers in a population with generally adequate Ca intake. In this cross-sectional analysis of 147 healthy women aged 31-43 years, fasting blood samples and three separate 24-h urinary samples were collected. Participants kept a 4-d food record and were divided into quartiles according to their dietary Ca:P ratios. The 1st quartile with Ca:P molar ratio < or = 0.50 differed significantly from the 2nd (Ca:P molar ratio 0.51-0.57), 3rd (Ca:P molar ratio 0.58-0.64) and 4th (Ca:P molar ratio > or = 0.65) quartiles by interfering with Ca metabolism. In the 1st quartile, mean S-PTH concentration (P = 0.021) and mean urinary Ca (U-Ca) excretion were higher (P = 0.051) than in all other quartiles. These findings suggest that in habitual diets low Ca:P ratios may interfere with homoeostasis of Ca metabolism and increase bone resorption, as indicated by higher S-PTH and U-Ca levels. Because low habitual dietary Ca:P ratios are common in Western diets, more attention should be focused on decreasing excessively high dietary P intake and increasing Ca intake to the recommended level.

Academic Dissertation by Virpi Kemi
Effects of dietary phosphorus and calcium-to phosphorus ratio on calcium and bone metabolism in healthy 20- to 43-year-old Finnish women
“This thesis contributes novel information related to the effects of dietary phosphorus (P) and the combined effects of P and calcium (Ca) on Ca and bone metabolism in healthy individuals. It is already well established that high P intake is detrimental for patients with impaired renal functioning, but the effects of high P intake in healthy humans have been investigated seldom. In this thesis, an excessively high P intake, which is common in Western countries, was observed to negatively affect Ca and bone metabolism. Moreover, P doses affected Ca and bone metabolism in a dose-dependent manner, and P sources also differed in their effects on the essential regulator of Ca and bone metabolism. Finally, we demonstrated that by increasing dietary Ca intake the negative effects of a high P intake could be reduced. However, based on the findings of the controlled study, even a high dietary Ca intake could not completely overcome all of the negative effects caused by a high dietary P intake.

Specific findings of each study were as follows:

Study I
In a controlled study with healthy young women, the oral intake of P in doses
comparable with normal dietary intakes (495, 745, 1245 and 1995 mg/d) with a low
Ca intake (250 mg/d) increased serum parathyroid hormone (S-PTH) concentration in
a dose-dependent manner. The highest P dose had the most negative effects, as with
the highest dose there was also a decrease in serum ionized Ca (S-iCa) concentration
and bone formation and an increase in bone resorption. Furthermore, a high P intake
(1995 mg/d) inhibited the increase in serum 1,25(OH)2D in response to a low dietary
Ca intake, implying that the normal relationship between Ca intake and Ca absorption
is disturbed in diets high in P and low in Ca. This study showed that P has a dose dependent effect on S-PTH and increases PTH secretion significantly when Ca intake
is low. An acutely high P intake adversely affects bone metabolism by decreasing bone
formation and increasing bone resorption.

Study II
In a controlled study of healthy 20-to 40-year-old women with a dietary P intake that was 3-fold above the dietary guidelines (1850 mg/d), by increasing the Ca intake from 480 mg/d to 1080 mg/d and then to 1680 mg/d, the S-PTH concentration decreased,
the S-iCa concentration increased and bone resorption decreased dose-dependently.
This study showed that a dietary Ca intake above the recommended level offers
several advantages in preventing the negative effects of a high P intake. However, not
even the highest Ca intake (1680 mg/d) could counteract the effect of high dietary P on
bone formation, as indicated by unchanged bone formation activity.

Study III
In a cross-sectional study with healthy 31- to 43-year-old women, a high habitual
dietary P intake was associated with increased S-PTH and decreased S-iCa concentrations. These results are in line with observations in our controlled short-term
study. Furthermore, in the habitual diets, phosphate additives, unlike other P sources,
were associated with higher mean S-PTH concentrations. The association of S-PTH
with natural P was the opposite of that with foods containing phosphate additives.
Thus, these results suggest that P sources might differ in their effects on the central
regulator of Ca and bone metabolism, parathyroid hormone.

Study IV
In a cross-sectional study of healthy 31- to 43-year-old women with an adequate Ca
intake, low habitual dietary Ca:P ratios (Ca:P molar ratio 0.50) were associated with
both higher S-PTH and U-Ca levels. Interestingly, the lowest Ca:P quartile with a Ca:P
molar ratio 0.50 differed from all other quartiles by having the most deleterious
associations with Ca metabolism. These results imply that a cut-off Ca:P ratio may
exist that is lower than the suggested Ca:P molar ratio of 1, below which the effects on
mineral metabolism and bone health are more severe. None of the study subjects
achieved the suggested dietary Ca:P molar ratio of 1.

A question arising from the findings of this thesis is how to reduce the current excessively high dietary P intake. The answer to this is not straightforward, as the current P intakes are not well known since the use of phosphate additives has not been taken into consideration when establishing food composition databases. In the future, food composition databases should be updated and the actual P contents of foodstuffs determined by laboratory analyses. In addition, it is impossible for an individual to know how much P foods contain because P is not included in the compulsory food ingredient list. While waiting for these issues to be addressed, one can reduce dietary P intake by restricting the consumption of highly processed foods and increasing the consumption of raw or unprocessed foods.

As there are still many open questions concerning the effects of high P intake on bone health, we will continue to investigate whether dietary P intakes and different P sources affect bone mass and structure in adult Finns. However, nowadays, when considering public health, it is not only osteoporosis but also other public health diseases in which a high P intake may play an important role in a negative sense. An alarming rise has been seen among Western populations in the incidence of type 2 diabetes, a major cause of end stage renal disease, in which dietary P restriction is a part of the treatment. Among patients with end-stage renal disease and diabetes, vascular calcification correlates highly with cardiovascular disease mortality. Recent results suggest that an excessive P intake may be involved in this vascular calcification process, even in healthy humans. Therefore, in the near future, our studies will expand to investigate the relationship between dietary P and vascular calcification in Finnish adults.” (pp. 85, 86)

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Source of Dietary Calcium: Chicken Egg Shell Powder

Also see:
Calcium Paradox
Hypertension and Calcium Deficiency
Blood Pressure Management with Calcium & Dairy
Calcium to Phosphorus Ratio, PTH, and Bone Health
Low CO2 in Hypothyroidism
Fatty Acid Synthase (FAS), Vitamin D, and Cancer
Parmigiano Reggiano cheese and bone health
Phosphate, activation, and aging

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.
Chicken eggshell powder (ESP) might be an attractive source of Ca for human nutrition. To study its nutritional value, we analyzed minerals, amino acids, and hormones in commercially available Slovakian ESP. The mineral composition was compared with three Dutch ESP samples that differed in feed and housing, a Japanese ESP, refined CaCO3, and an oyster shell supplement. Chicken eggshell powder contains high levels of Ca (mean +/- SD/g EPS: 401+/-7.2 mg) and Sr (372+/-161 microg) when compared with recommended or estimated daily intakes for humans 51 to 70 yr of age. Levels of potentially toxic Pb, Al, Cd, and Hg were very low as were levels of V, B, Fe, Zn, P, Mg, N, F, Se, Cu, and Cr. Large differences in the levels of F, Se, Cu, Cr, and Sr in the Dutch and Slovakian ESP indicated a strong influence of feed and environment. The small protein fraction of ESP contains high levels of Gly and Arg. Furthermore, small amounts of transforming growth factor-beta1 (0.75 to 7.28 ng/g ESP), calcitonin (10 to 25 ng/g ESP), and progesterone (0.30 to 0.33 ng/g ESP) were detected. Estradiol-17beta and calcitriol were below the detection limit of the methods used. Compared with ESP, refined CaCO3 was found to contain increased levels of Cd, and the oyster shell supplement showed increased levels of Al and Cd. Therefore, ESP seems to have a beneficial composition with about 39% of elemental Ca, relevant amounts of Sr, and low levels of Al, Pb, Cd and Hg. It may be used as a Ca source in human nutrition.

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.
Although bone metabolism is largely under genetic control, the role of nutrition is considerable. The present study evaluates the effects of chicken eggshell powder, a new source of dietary Ca, and purified CaCO3 on bone mineral density (BMD) of the lumbar spine and hip. Besides BMD we also looked at biochemical markers of bone and Ca metabolism. Both Ca sources were provided in combination with minerals and vitamins including Mg, cholecalciferol and phylloquinone. We designed a randomised, double-blind, placebo-controlled study to take place over 12 months. Healthy Caucasian women (n 85), selected by age (> or =50 and <70 years), from the databases of general practitioners were recruited by telephone calls. They had to be at least 5 years post-menopausal, with lumbar spine T-score being > – 2.5. At baseline, their mean habitual daily Ca intake was adequate. The women were randomly allocated to: eggshell powder-enriched (group A; n 24), purified CaCO3-enriched (group B; n 22), or a placebo product (group C; n 27). BMD was measured at baseline and then after 6 and 12 months of supplementation as were the biochemical markers bone-specific alkaline phosphatase, amino-terminal propeptide extension of type I collagen, deoxypyridinoline, calcitonin, intact parathyroid hormone, calcidiol, and urinary Ca. After 12 months of supplementation, only mean BMD of the femoral neck in group A was significantly increased (P=0.014) by 1.75% (95% CI 0.18, 3.32) compared with a decrease of -0.60% (95% CI -1.92, 0.72) in group C. This increase coincided with significant decreases in markers of bone resorption and formation. No significant changes were seen in BMD at other sites, including lumbar spine, nor in groups B and C. No differences were found between groups A and B, or B and C. The present study indicates that healthy late post-menopausal women with an adequate Ca intake at baseline may increase BMD of the hip within 12 months following supplementation with the chicken eggshell powder-enriched supplement.

Clin Calcium. 2005 Jan;15(1):95-100.
[Hen’s eggshell calcium].
[Article in Japanese]
Masuda Y.
In Japan, insufficient calcium (Ca) intake is serious problem for health which may be associated with the high prevalence of osteoporosis among the aged. The intake of most nutrients has been sufficient, however, the Ca intake has never been sufficient. Eggshell Ca has as much as 38% of Ca and low phosphorus content. Eggshell Ca was more soluble than Ca carbonate and was as much as milk products. Eggshell Ca has been shown to exhibit higher absorptivity and availability than Ca carbonate. Furthermore, it has been reported that eggshell Ca is more effective in increasing bone mineral density in ovariectomized osteoporotic rats. These results suggest that eggshell Ca could be beneficial for bone and we propose Ca fortified foods which contain eggshell Ca as a nutraceutical.

Int J Clin Pharmacol Res. 2003;23(2-3):83-92.
Eggshell calcium in the prevention and treatment of osteoporosis.
Rovenský J, Stancíková M, Masaryk P, Svík K, Istok R.
In this paper the most significant biological and clinical aspects of a biopreparation made of chicken eggshells are reviewed. Eggshell powder is a natural source of calcium and other elements (e.g. strontium and fluorine) which may have a positive effect on bone metabolism. Experimental and clinical studies performed to date have shown a number of positive properties of eggshell powder, such as antirachitic effects in rats and humans. A positive effect was observed on bone density in animal models of postmenopausal osteoporosis in ovariectomized female rats. In vitro eggshell powder stimulates chondrocyte differentiation and cartilage growth. Clinical studies in postmenopausal women and women with senile osteoporosis showed that eggshell powder reduces pain and osteoresorption and increases mobility and bone density or arrests its loss. The bioavailability of calcium from this source, as tested in piglets, was similar or better than that of food grade purified calcium carbonate. Clinical and experimental studies showed that eggshell powder has positive effects on bone and cartilage and that it is suitable in the prevention and treatment of osteoporosis.

Bratisl Lek Listy. 1999 Dec;100(12):651-6.
Short-term effects of a chicken egg shell powder enriched dairy-based products on bone mineral density in persons with osteoporosis or osteopenia.
Schaafsma A, Pakan I.
Based on the high calcium content, chicken egg shells are an interesting source of calcium. We studied the short-term effects on bone mineral density (BMD) of the lumbar spine and hip in 9 women and one man (mean age +/- SD, 63.9 +/- 8.1 years) with osteoporosis or osteopenia. Also the effects on pain and general well-being were monitored. Ten women (62.5 +/- 5.0 years) from a population study on BMD served as a control group. During a study period of 4-8 months, the intervention group consumed twice daily a dairy-based supplement which resulted in a daily intake of, among others, 3.0 g of egg shell powder, 400 IU of vitamin D3 and 400 mg of magnesium. BMD of the lumbar spine (anteroposterior (AP) and lateral (LA) position) and hip were measured by dual-energy X-ray absorptiometry. After the intervention period, BMDs of the lumbar spine, total proximal femur and trochanter were significantly (p < 0.05) increased with (median) 4.4%: (range) 1.7 to 10.4% (lumbar spine AP), 5.7%: -1.3 to 15.9% (lumbar spine LA), 2.2%: -1.9 to 9.4% (total proximal femur), 1.8%: -1.8 to 9.0% (trochanter). Within a period of 4 months, an important reduction in pain was reported and as a consequence an improvement in general well-being. In the control group, BMDs of the lumbar spine AP and of the femoral neck significantly decreased over a period of 8 months with -0.7% (-1.3 to 0.2%) and -0.9% (-2.4 to -0.1%) respectively. Six women of the intervention group continued to use the supplement on their own free will and without any check on compliance, up to 24 months. They consumed the supplement only once daily except for the last three months when they were asked to take the double dosage again. After 24 months BMDs did not differ from baseline. This study shows that egg shell powder is a source of bioavailable calcium. Furthermore, this pilot study indicates that the chicken egg shell powder enriched dairy-based supplement increases BMD of subjects with a low bone mass in the short term and as a consequence delays bone demineralisation for a longer period.

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Bone Health and Vitamin K

Also see:
Intestinal Serotonin and Bone Loss
Carbohydrates and Bone Health
Parmigiano Reggiano cheese and bone health
Calcium Paradox

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.
Vitamin K2 (MK4) functions were investigated by using the induced skin cell into osteoblast compared with the control media.The real time PCR measured gene expression in both cultures at the fourth, seventh, fourteenth, twenty first, twenty eighth, thirty fifth and forty second days of culture. The gene expressions of osteocalcin, osteonectin, osteopontin, bone sialoprotein, CBFA1, Interleukin-6, Estrogen receptors and collagen type) were monitored by real time PCR. MK4 had strong power to stimulate gene expression of osteocalcin and osteonectin after one week of culture but MK4 showed weak action on gene of osteopontin, bone sialoprotein and interleukin-6. The gene of estrogens showed the marked expression of estrogen receptor beta at the fourteenth day of culture while estrogen receptor alpha did not respond. MK4 could stimulate genes of RANKL and collagen type 1. This study supported the action of vitamin K2 for enhancing the bone matrix.

Nutrition. 2001 Oct;17(10):880-7.
Vitamin K and bone health.
Weber P.
In the past decade it has become evident that vitamin K has a significant role to play in human health that is beyond its well-established function in blood clotting. There is a consistent line of evidence in human epidemiologic and intervention studies that clearly demonstrates that vitamin K can improve bone health. The human intervention studies have demonstrated that vitamin K can not only increase bone mineral density in osteoporotic people but also actually reduce fracture rates. Further, there is evidence in human intervention studies that vitamins K and D, a classic in bone metabolism, works synergistically on bone density. Most of these studies employed vitamin K(2) at rather high doses, a fact that has been criticized as a shortcoming of these studies. However, there is emerging evidence in human intervention studies that vitamin K(1) at a much lower dose may also benefit bone health, in particular when coadministered with vitamin D. Several mechanisms are suggested by which vitamin K can modulate bone metabolism. Besides the gamma-carboxylation of osteocalcin, a protein believed to be involved in bone mineralization, there is increasing evidence that vitamin K also positively affects calcium balance, a key mineral in bone metabolism. The Institute of Medicine recently has increased the dietary reference intakes of vitamin K to 90 microg/d for females and 120 microg/d for males, which is an increase of approximately 50% from previous recommendations.

Clin Calcium. 2007 Nov;17(11):1752-60.
[Experience of vitamin K2 in Thailand].
[Article in Japanese]
Bunyaratavej N.
Vitamin K(2) has dual actions, stimulates osteoblastic functions, for synthesis of osteocalcin, osteonectin and other matrix bone proteins, in addition, new finding, in stem cell culture found osteoblast producing gene expression of collagen type 1, the other action, vitamin K(2) contains mild antiresorpion by inducing the osteoclastic apoptosis. Our study found that postmenopausal and elderly women have high risk of vitamin K(2) deficiency, comparing to the normal value of young, reproductive females. The efficacy of vitamin K(2) will be fulfill benefit after 6 months of administration, prolong use will enhance of bone quality that prevent fracture.

Clin Calcium. 2008 Oct;18(10):1476-82.
[Vitamin K2].
[Article in Japanese]
Ishida Y.
Vitamin K2 has been approved for the treatment of osteoporosis in Japan since 1995. Vitamin K2 treatment in osteoporosis has been shown to inhibit the occurrence of new bone fractures and to maintain BMD. The uniqueness of the prevention of bone fractures by vitamin K2 is that there has been no direct evidence of the relationship between increase of BMD and a decrease in the occurrence of bone fractures. A recent systematic review of seven Japanese randomized controlled trials by Cockayne has also shown that supplementation with phytonadione (Vitamin K1) and menaquinone (Vitamin K2) , particularly menaquinone-4, is associated with increased BMD and reduced fracture incidence. To confirm these results, a larger well design RCT using fractures as the primary endpoint is clearly needed.

Am J Clin Nutr January 1999 vol. 69 no. 1 74-79
Vitamin K intake and hip fractures in women: a prospective study
Diane Feskanich, Peter Weber, Walter C Willett, Helaine Rockett, Sarah L Booth and Graham A Colditz
Background: Vitamin K mediates the γ-carboxylation of glutamyl residues on several bone proteins, notably osteocalcin. High serum concentrations of undercarboxylated osteocalcin and low serum concentrations of vitamin K are associated with lower bone mineral density and increased risk of hip fracture. However, data are limited on the effects of dietary vitamin K.
Objective: We investigated the hypothesis that high intakes of vitamin K are associated with a lower risk of hip fracture in women.
Design: We conducted a prospective analysis within the Nurses’ Health Study cohort. Diet was assessed in 72327 women aged 38–63 y with a food-frequency questionnaire in 1984 (baseline). During the subsequent 10 y of follow-up, 270 hip fractures resulting from low or moderate trauma were reported.
Results: Women in quintiles 2–5 of vitamin K intake had a significantly lower age-adjusted relative risk (RR: 0.70; 95% CI: 0.53, 0.93) of hip fracture than women in the lowest quintile (<109 µg/d). Risk did not decrease between quintiles 2 and 5 and risk estimates were not altered when other risk factors for osteoporosis, including calcium and vitamin D intakes, were added to the models. Risk of hip fracture was also inversely associated with lettuce consumption (RR: 0.55; 95% CI: 0.40, 0.78) for one or more servings per day compared with one or fewer servings per week), the food that contributed the most to dietary vitamin K intakes.
Conclusions: Low intakes of vitamin K may increase the risk of hip fracture in women. The data support the suggestion for a reassessment of the vitamin K requirements that are based on bone health and blood coagulation.

Clin Calcium. 2005 Apr;15(4):605-10.
[Vitamin K2 as a protector of bone health and beyond].
[Article in Japanese]
Kaneki M.
Several lives of evidence indicate a protective effect of vitamin K against osteoporosis. Epidemiological studies showed that low vitamin K intake is associated with the increased risk of osteoporosis. Vitamin K2 (menatetrenone, MK-4) has been clinically used in the treatment of patients with osteoporosis in Japan, Korea and Thailand. Previous studies demonstrated the efficacy of vitamin K2 (45 mg/day) to prevent bone loss and reduce the rate of vertebral fractures, although a large, randomized intervention study is anticipated to provide more detailed evidence. Recently, vitamin K2 has been shown to reduce the progression of hepatocarcinoma. Moreover, it has been proposed that vitamin K may also have beneficial effects to prevent atherogenesis. The clarification of molecular mechanisms by which vitamin K2 exerts these salutary effects deserve further investigations.

Int J Gynaecol Obstet. 1997 Jan;56(1):25-30.
Serum vitamin K level and bone mineral density in post-menopausal women.
Kanai T, Takagi T, Masuhiro K, Nakamura M, Iwata M, Saji F.
OBJECTIVE:
Vitamin K is known to influence bone metabolism by facilitating the synthesis of osteocalcin (BGP). The bone mineral density decreases drastically after menopause. We investigated the relationship of bone mineral density, vitamin K levels and other biological parameters of bone metabolism in post-menopausal women.
METHODS:
Serum levels of vitamin K, BGP and other markers of bone metabolism were measured in 71 post-menopausal women (19 with reduced bone mineral density and 52 with normal bone density), and 24 women with climacteric symptoms receiving hormone replacement therapy (HRT), (6 with reduced bone mineral density and 18 with normal density).
RESULTS:
In the first group, women with reduced bone mineral density showed lower levels of vitamin K1 and K2 than those with normal bone mineral density. In the other group, the level of BGP decreased but levels of vitamin K showed no increase during HRT.
CONCLUSION:
The present findings suggested that vitamin K was related to post-menopausal bone mineral loss.

Annu Rev Nutr. 1995;15:1-22.
Role of vitamin K in bone metabolism.
Vermeer C, Jie KS, Knapen MH.
Vitamin K is a cofactor required for the formation of gamma-carboxyglutamate (Gla) residues in proteins. Osteoblasts produce at least three different Gla-containing proteins: osteocalcin, matrix Gla-protein, and protein S. After cellular secretion of these proteins, the main part of each remains bound to the hydroxyapatite matrix in bone, but their function remains unclear. Part of the newly synthesized osteocalcin is also set free into the bloodstream, where it may be used as a diagnostic marker for bone formation. Several studies have demonstrated that a poor vitamin K status is associated with an increased risk of osteoporotic bone fractures. Whether vitamin K supplementation will reduce the rate of bone loss in postmenopausal women remains a matter of debate.

J Gravit Physiol. 1998 Oct;5(2):65-9.
Bone markers during a 6-month space flight: effects of vitamin K supplementation.
Vermeer C, Wolf J, Craciun AM, Knapen MH.
Rapid bone loss is a serious health problem for astronauts during long lasting missions in space. We have recorded the changes of biochemical markers for bone metabolism in one of the astronauts during the 6-month space flight of the EUROMIR-95 mission. Immediately after launch both bone resorption markers and urinary calcium excretion increased about two fold, whereas bone formation markers remained unchanged. After 12 1/2 weeks the astronaut received vitamin K1 (10 mg/day for 6 weeks). Vitamin K is known to be involved in the formation of gamma-carboxyglutamate (Gla) in proteins, such as the calcium-binding bone Gla-proteins osteocalcin and matrix Gla-protein. Concomitant with the start of vitamin K treatment, the calcium-binding capacity of osteocalcin increased, and so did the urinary excretion of free Gla. This is suggestive for a subclinical vitamin K-deficiency in the astronaut before vitamin K-supplementation. During periods of high vitamin K status markers for bone formation (osteocalcin and bone alkaline phosphatase) had increased as compared to the first part of the flight. The mean increases were 14 and 23%, respectively. Our data suggest that increased intake of vitamin K may contribute to counteracting microgravity-induced loss of bone mass during long lasting space missions, but need confirmation in more astronauts.

Am J Clin Nutr. 2000 May;71(5):1201-8.
Dietary vitamin K intakes are associated with hip fracture but not with bone mineral density in elderly men and women.
Booth SL, Tucker KL, Chen H, Hannan MT, Gagnon DR, Cupples LA, Wilson PW, Ordovas J, Schaefer EJ, Dawson-Hughes B, Kiel DP.
BACKGROUND:
Vitamin K has been associated with bone mineral density (BMD) and risk of hip fracture. The apolipoprotein (apo) E4 allele (APOE*E4) has been associated with bone fracture through a putative effect on vitamin K transport in blood.
OBJECTIVE:
The objective was to determine the associations between vitamin K intake, apo E genotype, BMD, and hip fracture in a population-based cohort of elderly men and women.
DESIGN:
Dietary vitamin K intake was assessed with a food-frequency questionnaire in 335 men and 553 women (average age: 75.2 y) participating in the Framingham Heart Study in 1988-1989. Incidence of hip fractures was recorded from 1988 to 1995. BMD at the hip, spine, and arm was assessed on 2 separate occasions (1988-1989 and 1992-1993). Comparisons between apo E genotype and BMD were made relative to E4 allele status (at least 1 epsilon4 allele compared with no epsilon4 allele).
RESULTS:
Individuals in the highest quartile of vitamin K intake (median: 254 microg/d) had a significantly lower fully adjusted relative risk (0.35; 95% CI: 0. 13, 0.94) of hip fracture than did those in the lowest quartile of intake (median: 56 microg/d). There were no associations between vitamin K intake and BMD in either men or women. No association was found between the E4 allele and BMD, and there were no significant interactions between the E4 allele and phylloquinone intake and BMD or hip fracture.
CONCLUSIONS:
Low vitamin K intakes were associated with an increased incidence of hip fractures in this cohort of elderly men and women. Neither low vitamin K intake nor E4 allele status was associated with low BMD.

Am J Clin Nutr. 2003 Feb;77(2):512-6.
Vitamin K intake and bone mineral density in women and men.
Booth SL, Broe KE, Gagnon DR, Tucker KL, Hannan MT, McLean RR, Dawson-Hughes B, Wilson PW, Cupples LA, Kiel DP.
BACKGROUND:
Low dietary vitamin K intake has been associated with an increased risk of hip fracture in men and women. Few data exist on the association between dietary vitamin K intake and bone mineral density (BMD).
OBJECTIVE:
We studied cross-sectional associations between self-reported dietary vitamin K intake and BMD of the hip and spine in men and women aged 29-86 y.
DESIGN:
BMD was measured at the hip and spine in 1112 men and 1479 women (macro x +/- SD age: 59 +/- 9 y) who participated in the Framingham Heart Study (1996-2000). Dietary and supplemental intakes of vitamin K were assessed with the use of a food-frequency questionnaire. Additional covariates included age, body mass index, smoking status, alcohol use, physical activity score, and menopause status and current estrogen use among the women.
RESULTS:
Women in the lowest quartile of vitamin K intake (macro x: 70.2 microg/d) had significantly (P < or = 0.005) lower mean (+/- SEM) BMD at the femoral neck (0.854 +/- 0.006 g/cm(2)) and spine (1.140 +/- 0.010 g/cm(2)) than did those in the highest quartile of vitamin K intake (macro x: 309 microg/d): 0.888 +/- 0.006 and 1.190 +/- 0.010 g/cm(2), respectively. These associations remained after potential confounders were controlled for and after stratification by age or supplement use. No significant association was found between dietary vitamin K intake and BMD in men.
CONCLUSIONS:
Low dietary vitamin K intake was associated with low BMD in women, consistent with previous reports that low dietary vitamin K intake is associated with an increased risk of hip fracture. In contrast, there was no association between dietary vitamin K intake and BMD in men.

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Parmigiano Reggiano cheese and bone health

Also see:
Intestinal Serotonin and Bone Loss
Carbohydrates and Bone Health
Bone Health and Vitamin K
Calcium Paradox
Nutritional Values – Parmigiano Reggiano

Clin Cases Miner Bone Metab. 2011 Sep;8(3):33-6.
Parmigiano Reggiano cheese and bone health.
Pampaloni B, Bartolini E, Brandi ML.
Osteoporosis is a multifactorial disease characterized by loss of bone mass and microarchitectural deterioration of bone tissue, which leads to a consequent increase in the risk of skeletal fractures. Diet awakes a critical interest in osteoporosis, because it is one of the few determinants that can be safely modified. A healthy well balanced nutrition can play an important role in prevention and pathogenesis of osteoporosis, but also in support of a pharmacological therapy. Numerous evidences have already established that dietary calcium, proteins and vitamin D are essential nutrients for achieved peak bone mass and maintaining skeletal health. Dairy products, by providing both calcium and proteins, represent the optimal source of highly bioavailable nutrients for bone health. Among dairy foods in particular cheese results one of the major source of calcium in the adults western diet and also in the Italian adults diet. Parmigiano Reggiano cheese is an homemade Italian food whose denomination “Protected Designation of Origin” is linked to an artisanal manufacturing process in limited geographic area of Northern Italy and is an optimal source of essential nutrients for acquisition and maintenance of bone health. Parmigiano Reggiano is a cheese easy digested, for the presence of ready to use proteins and lipids, lactose free, rich in calcium, with possible prebiotic and probiotic effect. On the basis of its nutritional characteristics and of its easy digestibility Parmigiano Reggiano cheese is recommended in all feeding age groups.

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