• Journal of Preventive Medicine and Public Health
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• v.44 no.1
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• pp.9-13
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• 2011

Objectives: Although increased reactive oxygen species (ROS) is caused by stress accelerates collagen degradation, there was no data on the relationship between stress and urinary hydroxyproline (Hyp) and proline (Pro), a good marker of collagen degradation. The purpose of this study was to evaluate the relationship between depression, anxiety, and stress (DAS) and concentrations of urinary Hyp and Pro. Methods: 97 hospital employees aged 20 to 58 were asked to fill out comprehensive self-administrated questionnaires containing information about their medical history, lifestyle, length of the work year, shit-work and DAS. Depression Anxiety Stress Scale (DASS) was applied to evaluate chronic mental disorders. Urine samples were analyzed using High Performance Liquid Chromatography (HPLC) with double derivatization for the assay of hydroxyproline and proline. Results: The mean value of Hyp and Pro concenturation in all subjects was $194.1{\pm}113.4\;{\mu}mol/g$ and $568.2{\pm}310.7\;{\mu}mol/g$. DASS values and urinary Pro concentrations were differentiated by sex (female > male, p < 0.05) and type of job (nurse > others, p < 0.05). In the stepwise multiple linear regressions, urinary Hyp and Pro concentrations were influenced by stress (Adjusted $r^2$ = 0.051) and anxiety and job (Adjusted $r^2$ = 0.199), respectively. Conclusions: We found that stress and anxiety were correlated with urinary Hyp and Pro concentrations. To identifying a definite correlation, further study in large populations will be needed.

• Journal of Nutrition and Health
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• v.29 no.9
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• pp.1013-1020
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• 1996

The purpose of this study was to investigate nutrient intake and bone status in rural postmenopausal women in Korea. Ten postmenopausal women in An-Sung area participated in this study and they were divided into two groups ; women in group I had been postmenopausal for 4 years or less and those in group II had been postmenopausal for 5 years or more before the present study began. Their environmental factors and dietary intakes were surveyed through the personal interviews. Serum levels of calcium, phosphorus, parathyroid hormone(PTH), estradiol and urinary Ca, P, creatinine, hydroxyproline levels were measured from December 4 to December 27 in 1993. The reults of this study are summarized as follows : Average ages of group I and II were 54.8 and 57.2 years. Average menopausal ages of group I and II were 50.8 and 47.3 years. The nutrient intakes of subjects were higher than recommended dietary allowances(RDA) except calorie, protein, calcium, and vitamin A. The nutrient status did not show any significant difference between group I and II. Serum levels of Ca, P, PTH, estradiol and urinary P, creatinine excretion did not show any significant difference between group I and II, and all levels were in normal range. Urinary Ca excretion(p<0.05) and hydroxyproline excretion(p<0.01) were significantly lower in group I than in group II. Urinary Ca/creatinine(Ca/cr) and hydroxyproline/creatinine(Hpr/cr) rations were significantly higher in group II than in group I(p<0.01. And Hpr/cr levels of group I were in normal range, but most of subjects in group II were higher than 0.017 indicating sign of osteoporosis. Correlations between parameters showed that serum PTH adn urinary Ca, Ca/cr levels were positive related (p<0.01), and the years of the after menopausal year and urinary Hpr/cr was also positive related(p<0.05). The present results suggests that it is difficult to protect postmenopausal women's bone destruction having Korean usual diets. Therefore, to prevent osteoporosis with aging, minimizing the hormonal changes in postmenopausal women is needed as well as Ca supplementation and proper exercise before menopause begins.

• Journal of Nutrition and Health
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• v.26 no.8
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• pp.915-924
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• 1993

To study the effect of menopause and dietary protein level on Ca metabolism, ovariectomy (OVAX) and sham operations were performed in 16-week old female rats. Each treatment group was fed for 16 weeks either 5%(L) or 50%(H) casein diets, forming SH, SL, OH, OL groups. High protein groups(SH, OH) showed higher Ca and hydroxyproline excretion in urine. Urinay hydroxyproline was also higher in OVAX, which tells the possibility of increased bone resorption by OVAX and by high dietary protein. At 16th wee, however, urinary Ca and hydroxyproline of SH caught up with OH group, whereas those of OL remained higher than SL. Therefore it seems that high dietary protein overrides the effect of OVAX with time. Urnary protein measured at 8th week was higher in high protein groups, especially in OH. GFR was not differ significantly among groups at 8th week. At 16th week, however, high protein groups showed twice the GFR value of low protein groups. Therefore the increase of urinary Ca and hydroxyproline in SH and OH groups can be explained partly by the increased GFR. The tendency of increased GFR and urinary excretion of protein, Ca, and hydroxyproline was most obvious in OH group. It seems that the effect of high protein diet is likely to accelerated by ovariectomy. The effect of Ovax and dietary protein on the composition of femur, scapular, and lumbar bones, was not pronounced. However, when only the high protein groups were compared, OVAX resulted in the reduction of bone weight, ash and Ca contents, especially in femur. The reason that was no significant effect on bone might be due to the short experimental period to induce that was no significant effect on bone might be due to the short experimental period to induce the changes on bone composition and dietary Ca content used in this experiment may have been high enough to prevent bone loss.

• Journal of Nutrition and Health
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• v.28 no.5
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• pp.415-425
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• 1995

This study was performed to investigate the effect of dietary protein level on the metabolic changes of Ca and skeletons in postmenopausal women, using ovariecotomized rats as an animal model. The female rats of 200∼250g were fed either 8%(L) or 50%(H) casein diet for 15 weeks(1st experiment). At 15th week, the rats of each diet group were undergone ovariectomy or sham-operation and they were continued to feed the same experimantal diet for 9 more months(2nd experiment). Ca metabolism, kidney function and bone composition were determined at the end of 1st experiment, 3rd and 9th month of 2nd experiment. After 1st experiment, high protein group showed higher urinary Ca and protein excretion, however, there was no difference in GFR and urinary hydroxyproline excretion. The weights, ash and Ca content of femur, scapular and vertebra tended to be higher in high protein groups which tells that high protein promoted skeletal growth. In 2nd experiment, high protein group showed higher urinary Ca and protein excretion and lower Ca absorption and balance. GFR was not affected by dietary protein and ovariectomy but increased with time, as well as kidney weight which shows the continuous development of kidney at this age of 15 month in rats. There were no difference in urinary hydroxyproline, serum ALP, and PTH among experimental groups. The weights of femur, scapular, 4th vertebra increased with time, showing the skeleton continues to grow at this age in rats. However, Ca contents, Ca/wt, Ca/ash were decreased with time and tended to be lower in high protein group especially in femur. In conclusion, prolonged feeding of high protein diet deteriorated Ca metabolism and induced bone loss as time after menopause is extended.

• Journal of Nutrition and Health
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• v.28 no.4
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• pp.298-308
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• 1995

To investigate the effect of estrogen and dietary protein level on Ca metabolism, female rats were undergone ovariectomy or sham-operation. Ovariectomized rate were divided into either estrogen-or vehicle-treated groups. Each treatment group was again divided into 40%-casein(H) or 10%-casein(L) diet groups. All experimental diets contained 0.2% Ca, 0.4% P and fed to rats for 8 weeks. Apparant Ca absorption and Ca balance were not affected by dietary protein level and ovariectomy, however they were increased by estrogen injection and this effect was even higher in low protein groups. Urinary Ca excretion were higher in high protein groups. GFR was not affected by dietary protein level, ovariectomy, or by estrogen injection. Urinary protein excretion was higher in high protein groups, which implies that the kidney funtion was deteriorated by high protein diet, and this may account partly for the higher urinary Ca in high protein groups. Ovariectomy or estrogen treatment had no effect on urinary protein excretion. Urinary hydroxyproline was higher in ovariectomized rats and increased in high protein grous. Elevated value of ovarictomized rats was lowered by estrogen injection, especially in low protein group. Alkaline phosphatase tended to increase in ovariectomized groups and lowered with estrogen treatment, but this difference was not statistically significant. Serum PTH was not affected by ovariectomy and dietary protein level. Therefore the increased hydroxproline excretion does not seem to be attributed to PTH. Dietary protein level, ovariectomy and estrogen treatment did not affect the weights and components of femur, scapular, and 4th vertebra. Ash/wt ratio of femur was, however, lower in ovariectomized rats and increased with estrogen treatment. Therefore, among the bones studied, femur seemed to be the most vulnerable. The results of this study shows that estrogen treatment may alleviate or reduce bone loss in postmenopausal women somewhat, especially for those people with low protein diet.

• Journal of Preventive Medicine and Public Health
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• v.35 no.4
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• pp.282-286
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• 2002

Objectives : The level of 4-hydroxyproline (4-Hyp) in human urine was measured using high performance liquid chromatography (HPLC) with a fluorescence detector. This method is useful for medical examinations and investigating the radicals induced by physical, chemical, mental stresses. This method is superior to many published several methods in terms of its low cost and ability to analyze many samples. Methods : The urine from workers in a tire manufacturing company (22 male pre- and post-shift workers) and 18 office-workers as controls were analyzed. Data concerning age, the cumulative drinking amount and the cumulative smoking amount was collected with a questionnaire. The optimum applied amount of dansyl-Cl, the optimum reaction temperature and time, the recoveries and the optimum pH of the eluent and buffer were determined.4-Hyp from human urine was derivatized with dansyl-Cl (dimethylamino-naphthalene-1-sulfonyl chloride) after removing the a-amino acid by a treatment with phthalic dicarboxaldehyde (OPA) and cleaned with Bond Elut C18 column. The 4-Hyp derivatives were separated on a reversed phase column by gradient elution with a phosphate buffer (5 mmol, pH 8.0) and acetonitrile, and detected by fluorescence measurements at 340 nm (excitation) and 538 nm (emission). Results : The detection limit for the urinary free 4-Hyp was $0.364{\mu}mol/l$. The recovery rate of 4-Hyp was 99.7%, and the effective pH of the phosphate buffer and borate buffer were 3.0 and 8.0, respectively. From statistical analysis, age, drinking and smoking did not affect the urinary free 4-Hyp in both the controls and workers. The range of urinary 4-Hyp in the controls, pre-shift, and post-shift workers were 0.33-16.44, N.D-49.06, and $0.32-56.27{\mu}mol/l$. From the pared-sample t-test, the urinary 4-Hyp levels in post-shift workers ($11.82{\pm}6.73\;nmmol/mg\;Cre$) were 2-fold higher than in pre-shift workers ($5.36{\pm}5.53\;nmol;/mg\;Cre$) and controls ($4.91{\pm}4.89\;nmol;/mg\;Cre$). Conclusions : This method was developed with high sensitivity, accuracy, and precision. The present method was effectively applied to analyze the urinary free 4-Hyp in both controls and workers.

• YAKHAK HOEJI
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• v.43 no.4
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• pp.526-534
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• 1999

We investigated the effect of safflower-seed on fracture healing of fracture model in rat. Fracture healing was evaluated by examining the degree of wound healing macroscopically, radiography, bone histomorphometry and biochemical examination. After 1, 3, 5, 7 days, the would healing was accelerate in safflower-seed diet group. Radiography does not reveal the difference in fracture healing between two group. After 2 weeks, safflower-seed had a significant, stimulatory effect on external callus formation (p<0.05). But after 4, 6, 8 weeks, no difference was observed between normal and safflower-seed dietgroup in callus size. Urinary hydroxyproline, osteocalcin and total alkaline phosphatase decreased significantly (p<0.05) in safflower-seed treated group at 2 week after tracture.

• Korean Journal of Community Nutrition
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• v.1 no.1
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• pp.71-78
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• 1996

This study was conducted to compare the dietary factors which influence on the bone status of 28 women in urban and 30 women in rural area. Urinary excretion of hydroxyproline(Hpr) and Calcium(Ca) were measured as biological markers of bone resorption. Mean daily intake levels of total protein, animal protein, total calcium, calcium, calcium from milk and milk products, animal calcium, Ca / P ratio by 24 hr recall method were significantly higher in urban women. However, mean daily sodium(Na) intake levels were not significantly different between two groups. Ca Index score and Na Index score by food frequency methods were also significantly higher in urban than in rural subjects. While urinary Ca excretion elves of two groups were similar, Na excretion levels were significantly higher in rural women. Mean urniary levels of Ca / creatinine(cr) and Hpr / cr as bone status index were within normal range and not significantly different between two groups. However, prevalence of poor bone status as assessed by hydroxyproline was higher in rural women. Na Index, urinary Ca excretion and Ca / cr ratio were significantly correlated with bone status(Hpr / cr) in urban women, while only age was related to bone status in rural women. These demonstrated that high Na intake results in increased urinary excretion of Na and Ca and could cause bone resorption. Multiple regression analysis indicated that Na Index score and age have greater effect than other variables in urban women and only age has greater effect in rural women.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.4
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• pp.657-661
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• 2001

Osteoporosis that is associated with ovarian hormone deficiency following menopause (postmenopausal osteoporosis) is by far the most common cause of age-related bone loss. Isoflavone has been reported as a natural substance that possibly minimizes bone loss in postmenopausal women. This study was conducted to investigate the preventing, treating effects of isoflavone on bone loss in ovariectomized rats. 120 Sprague Dawley rats of 13 week-old were devided into 2 groups, a treatment group and prevention group. Each group was consisted of six subgroups; control (CON), sham operated (SH) or ovariectomized (OVX) and isoflavone supplemented goups: OVX+0.25mg isoflavone/kg diet (OL), OVX+0.8mg isoflavone/kg diet(OM) and OVX+2.5mg isoflavone/kg diet(OH). to study the preventing effects of isoflavone on bone loss, OL, OM and OH groups were fed with isoflavone from 4 days after ovariectomization. Treating effects of isoflavone on bone metabolism were investigated with OL, OM, OH groups supplemented with isoflavone from 8 weeks after ovariectomization. Isoflavone supplementation continued for 8 weeks. At 8 weeks after ovariectomization significant increase in alkaline phosphatase occurred comparing with CON and SH group. By isoflavone supplementation from 4 days after ovariectomy alkaline phosphatase and urinary hydroxyproline were lowered and bone mineral density, bone strength of the femur and tibia and bone dry weight were slightly enhanced with no significant difference. Isoflavone supplemented group at the level of 0.8mg/kg diet (OM group) had significantly lower serum alkaline phosphatase, urinary hydroxyproline, and higher strength of femur than OVX group. Groups with isoflavone supplementation fro 8 weeks after ovariectomy had lower level of serum alkaline phosphatase, urinary hydroxyproline than OVX group. Bone mineral density, bone dry weight and bone strength of the femur and tibia were slightly enhanced by isoflavone supplementation. However there was no significanct difference between OVS ad isoflavone supplementation groups. The results suggest that isoflavone might have potential role for preventing postmenopausal bone loss. Isoflavone supplementation at early stage of postemenopause may be beneficial to age-related bone health.

• Journal of Nutrition and Health
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• v.22 no.6
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• pp.497-506
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• 1989

To investigate the effect of levels of dietary protein and age on bone metabolism 40% and 5% casein were fed to the rats of 2 & 13 months of age for 12 weeks. High protein groups showed higher bone weight and Ca content than low protein groups and urinary Ca loss was increased in high protein groups but the difference disappeared gradually. A significant increase in urinary hydroxyproline excretion was noted in high protein groups of both age. Another short term study was undertaken to study if the above effect was related with renal function or PTH. Extremely high and low protein diets(60%, 6%) were fed to the rats of different ages(6wks, 6mos.) for 2 weeks, Urinary Ca excretion was significantly increased in high protein groups of young and aged rats and GFR was increased as well. There was no difference in serum iPTH levels between low and high protein groups, but it was elevated in aged rats. Alkaline phosphatase activity was higher in young rats, reflecting faster bone formation. The observed hypercalciuria in high protein groups, especially in aged rats, seems to be related to higher GFR, and PTH dose not appear to be a major mediator.