• Korean Chemical Engineering Research
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• v.58 no.1
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• pp.106-112
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• 2020

Analyzing vitamin D levels is important for monitoring health conditions because vitamin D deficiency is associated with various diseases such as rickets, osteomalacia, cardiovascular disorders and some cancers. However, vitamin D concentration in the blood is very low with optimal level of 75 nmol/L, making quantitative analysis difficult. The objective of this study was to develop a highly sensitive analysis method for vitamin D using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-MS). 25-hydroxyvitamin D (25(OH)D), which has been used as an indicator of vitamin D metabolites in human biofluids was chemically derivatized using a secosteroid signal enhancing tag (SecoSET) with powerful dienophile and permanent positive charge. The SecoSET-derivatized 25(OH)D provided good linearity (R² > 0.99) and sensitivity (limit of quantitation: 11.3 fmol). Chemical derivatization of deuterated 25-hydroxyvitamin D₃ (d₆-25(OH)D3) with SecoSET enabled absolute quantitative analysis using MALDI-MS. The highly sensitive method could be successfully applied into monitoring of quantitative changes of bioactive vitamin D metabolites after treatment with ketoconazole to inhibit 1α-hydroxylase reaction related to vitamin D metabolism in human breast cancer cells. Taken together, we developed a MALDI-MS-based platform that could quantitatively analyze vitamin D metabolites from cell products, blood and other biofluids. This platform may be applied to monitor various diseases associated with vitamin D deficiency such as rickets, osteomalacia and breast cancer.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.17
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• pp.7337-7341
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• 2014

Background: The physiological role of vitamin D extends beyond bone health and calcium-phosphate homeostasis to effects on cancer risk, mainly for colorectal cancer. Vitamin D may have an anticancer effect in colorectal cancer mediated by binding of the active form $1,25(OH)_2D$ to the vitamin D receptor (VDR). The Taq1 VDR gene polymorphism, a C-to-T base substitution (rs731236) in exon 9 may influence its expression and function. The aim of this study wass to determine the 25(OH)D vitamin D level and to investigate the association between circulating vitamin D level and Taq1VDR gene polymorphism among Jordanian colorectal cancer patients. Materials and Methods: This case control study enrolled ninety-three patients and one hundred and two healthy Jordanian volunteers from AL-Basheer Hospital/Amman (2012-2013). Ethical approval and signed consent forms were obtained from all participants before sample collection. 25(OH)D levels were determined by competitive immunoassay Elecsys (Roche Diagnostic, France). DNA was extracted (Promega, USA) and amplified by PCR followed by VDR Taq1 restriction enzyme digestion. The genotype distribution was evaluated by paired t-test and chi-square. Comparison between vitamin D levels among CRC and control were assessed by odds ratio with 95% confidence interval. Results: The vitamin D serum level was significantly lower among colorectal cancer patients (8.34 ng/ml) compared to the healthy control group (21.02ng/ml). Patients deficient in vitamin D (less than 10.0 ng/ml) had increased colorectal cancer risk 19.2 fold compared to control. Only 2.2% of CRC patients had optimal vitamin D compared to 23.5% among healthy control. TT, Tt and tt Taq1 genotype frequencies among CRC cases was 35.5%, 50.5% and 14% compared to 43.1%, 41.2% and 15.7% among healthy control; respectively. CRC patients had lower mean vitamin D level among TT ($8.91{\pm}4.31$) and Tt ($9.15{\pm}5.25$) genotypes compared to control ($21.3{\pm}8.31$) and ($19.3{\pm}7.68$); respectively. Conclusions: There is significant association between low 25(OH)D serum level and colorectal cancer risk. The VDRTaq1 polymorphism was associated with increased colorectal cancer risk among patient with VDRTaq1 TT and Tt genotypes. Understanding the functional mechanism of VDRTaq1 TT and Tt may provide a strategy for colorectal cancer prevention and treatment.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.3
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• pp.408-415
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• 2013

The objective of this experiment was to assess the use of different vitamin D metabolites in the feed of broiler chickens and the effects of the metabolites on performance, bone parameters and meat quality. A total of 952 one-day-old male broiler chicks were distributed in a completely randomised design, with four treatments, seven replicates and 34 birds per experimental unit. The treatments consisted of four different sources of vitamin D included in the diet, $D_3$, $25(OH)D_3$, $1,25(OH)_2D_3$, and $1{\alpha}(OH)D_3$, providing 2000 and 1600 IU of vitamin D in the starter (1 to 21 d) and growth phases (22 to 42 d), respectively. Mean weight, feed:gain and weight gain throughout the rearing period were less in animals fed $1{\alpha}(OH)D_3$ when compared with the other treatments (p<0.05). No significant differences were noted among the treatments (p>0.05) for various bone parameters. Meat colour differed among the treatments (p>0.05). All of the metabolites used in the diets, with the exception of $1{\alpha}(OH)D_3$, can be used for broiler chickens without problems for performance and bone quality, however, some aspects of meat quality were affected.

• Journal of Nutrition and Health
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• v.45 no.1
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• pp.20-29
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• 2012

Metabolic syndrome (MetS) has become a global epidemic. In particular, it is known that there is a dramatic increase in the prevalence of MetS among women during the postmenopausal period. Recently, accumulating studies have suggested that vitamin D deficiency may be inversely associated with the risk factors regarding MetS. However, evidence from postmenopausal women is limited. In this study, we examined the association between the serum 25-hydroxyvitamin D [25(OH)D] and the MetS in Korean adult women aged 20-69 years (n = 2,618) by using the 2007-2008 Korean National Health and Nutrition Examination Survey data. The geometric mean of plasma 25(OH)D were $17.16{\pm}6.28\;ng$/mL and $20.20{\pm}7.69\;ng$/mL for premenopausal and postmenopausal women, respectively. The percentages of vitamin D deficiency [25(OH)D < 12 ng/mL] were 22.5% and 14.4%, respectively. MetS was more prevalent in postmenopausal women (43.0%) compared with premenopausal women (11.2%). When serum concentrations of 25(OH)D were categorized in quintiles, there was no relationship in the prevalence of MetS in both premonopausal and postmenopausal women. However, in premenopausal women, compared with the lowest 25(OH)D quintile, the odds ratio for hypertriglyceridemia in the highest quintile was 0.57 (95% CI 0.34-0.95, $P_{trend}$ = 0.041) and for low serum HDL cholesterol 0.60 (95% CI 0.42-0.85, $P_{trend}$ = 0.014) after adjusting for all potential confounders. On the other hand, we observed the tendency of an inverse relationship for 25(OH)D regarding low serum HDL cholesterol (OR 0.78, 95% CI 0.50-1.22, $P_{trend}$ = 0.029) and a direct relationship with abdominal obesity (OR 1.94, 95% CI 1.01, 3.74, $P_{trend}$ = 0.049) in postmenopausal women. Further studies are needed to confirm these findings in other research settings.

• The korean journal of orthodontics
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• v.27 no.2
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• pp.335-347
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• 1997

Vitamin D is known to exert its action by activating DNA and RBA within target cells to produce proteins and enzymes that can be used in bone resorption process. Particularly, the active form of vitmain D, 1,25-dihydroxycholecalciferol $[1,25-(OH)_2D_3]$, is considered to be one of the most potent stimulators of osteoclatic acitivity in vitro. The purpose of this study was to evaluate the effect of 1,25-Dihydroxyvitamin $D_3$ on the avtivity of periodotal ligament cells and, the experimental tooth movement. Human periodontal ligament cells were collected from the first premolar tooth extracted for the orthodontic treatment, and were incubated in the environment of $37^{\circ}C$, 5% $CO_2$ and 95% humidity. Microtitration(MIT) assay was done at 10, 25, 50 and 100ng/ml of 1,25-Dihydroxyvitamin $D_3$. 21 Sprague-Daft rats were divided into a control gmup(3), and experimental groups(18) where 100g of force from helical spring was applied across the maxillary incisors 1,25-Dihydroxyvitamin $D_3$ was injected into periodontal ligament at the mesial or distal surface of maxillary incisors so that we can compare the control side and the experimental side. Expreimental groups were sac rifled at 12, 24, 36, 48, 72hours and 7 days after force application, respectively. And the obtained tissues were evaluated histologically. The observed results were as follows. 1. The activity of periodontal ligament cells in l0ng/ml or 25ng/ml of 1,25-Dihydroxyvitamin $D_3$ 1,25-Dihydroxyvitamin $D_3$ was not significantly different to the control at the cultivation of 1, 2 and 3 days. 2. The activity of periodontal ligament cells was significantly increased at 3 days in 50 ng/ml of 1,25-Dihydroxyvitamin $D_3$ and 2, 3 days in 100g/ml of 1,25-Dihydroxyvitamin $D_3$. 3. Up to 7 days after force application, there was no difference in osteoblastic activity, tearing of periodontal ligament and proliferation of capillary at tension side between 1,25-Dihydroxyvitamin $D_3$ injection side and the control side. 4. The osteoclastic activity and the resorption of alveolar bone was greater in 1,25-Dihydroxyvitamin $D_3$ injection side than the control side at 36 hours after force application.

• Journal of Animal Science and Technology
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• v.45 no.3
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• pp.455-462
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• 2003

A study was conducted to determine the effects of feeding a diet containing no Ca supplement (limestone) during the late finishing period on growth, marbling and serum 1,25-dihydroxy vitamin $D_3$ level in Korean native cattle. Twenty-four steers (20${\sim}$24 mo of age) were divided into two groups of 12 each: one group assigned to a control diet (concentrates containing 2.5% limestone) and the other to a diet containing no calcium supplement. They were allowed to have free access to diets (concentrates and orchard grass hay) and water during the entire feeding period (223 d). Serum $Ca^{2+}$, Ca and P concentrations were not influenced by diets, but serum 1,25-dihydroxy vitamin $D_3$ concentrations determined 2 or 6 mo after the beginning of feeding the experimental diets were higher (P<0.01) in steers fed the diet without Ca supplement than in those fed the control diet (78.3 vs 51.7 and 80.3 vs 51.1 pg/mL, respectively). Steers fed the diet without Ca supplement tended to have a higher intake of concentrates, but a lower intake of hay, compared to those fed the control diet. Average daily gain was higher (P<0.05) in steers fed the diet without Ca supplement than in those fed the control diet. Feeding the diet without Ca supplement remarkably (P<0.01) increased the marbling score (5.1 vs 2.2) and the muscle (M. longissimus dorsi) fat content (10.2 vs 6.7%) with a concomitant decrease in moisture content (67.6 vs 70.4%), compared to feeding the control diet. Ribeye area was increased (77.2 vs 82.8 $cm^2$) with the diet without Ca supplement, compared to the control diet (P<0.05). Meat color, pH and water-holding capacity in longissimus muscle were not different between the two groups. The Warner-Brazler Shear (WBS) force of the longissimus muscle was slightly (P=0.08) lower in steers fed the diet without Ca supplement than in steers fed the control diet (2.9 vs 3.2 kg/1.27-cm diameter core). Sensory evaluation showed that feeding the diet without Ca supplement slightly (P<0.05) improved tenderness (4.9 vs 4.5) and flavor (4.9 vs 4.6), compared to feeding the control diet, but juiciness was not affected by diets. Results showed that deletion of Ca supplement from finishing diets is beneficial, increasing growth and marbling partly through an increased energy intake and induced 1,25-dihydroxy vitamin $D_3$ synthesis that may increase intracellular $Ca^{2+}$ concentration and in turn fat synthesis.

• The Journal of Korean Academy of Prosthodontics
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• v.46 no.6
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• pp.620-627
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• 2008

STATEMENT OF PROBLEM: A biochemical approach for surface modification has offered an alternative for physicochemical and morphological methods to obtain desirable bone-implant interfaces. PURPOSE: The purpose of the present study was to investigate cell responses to poly (D,L-lactide-co-glycolide) (PLGA)/$1{\alpha}$,25-(OH)$_2D_3$ coating with reference to cellular proliferation and differentiation in vitro. MATERIAL AND METHODS: 96 titanium discs were fabricated and divided into four groups. Group 1 was anodized under 300 V as control. Group 2, 3 and 4 were anodized then coated with 3 ml PLGA/$1{\alpha}$,25-(OH)$_2D_3$ solutions. Amount of the solutions were 2 ul, 20 ul and 200ul respectively. The osteoblast-like Human Osteogenic Sarcoma (HOS) cells were seeded and cultured for 1, 3 and 7 days. MTSbased cell proliferation assay and ALPase activity test were carried out. RESULTS: PLGA nanoparticles were observed as fine, smooth and round and HOS cells attached to the anodized surfaces through strand-like and sheet-like filopodia. After 3 days of culture, the dendritic filopodia were exaggerated and sheet-like cytoplasmic projections covered the coated titanium surfaces. After 3 days of culture, all of the groups showed increased cellular proliferation and the lowest proliferation rate was measured on group 2. Higher amount of incorporated $1{\alpha}$,25-(OH)$_2D_3$ (Group 3 and 4) improved cellular proliferation but the differences were not significant statistically (P > .05). But they increased the rate of ALP activities than the control group at day 3 (P < .05). CONCLUSION: Biodegradable PLGA nanoparticles incorporated with vitamin D metabolite positively affected proliferation and differentiation of cells on the anodized titanium surface.

• The korean journal of orthodontics
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• v.25 no.6 s.53
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• pp.705-714
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• 1995

Due to the great deal of effort that has gone into the study of osteoclastic differentiation and activation over the last few decades, the mechanisms of these two events have been discovered gradually. Nitric oxide($NO^-$), which is produced from arginine by a nitric oxide synthase, opened up a new area of biological research. Recently, it has been reported that $NO^-$ is produced by osteoblasts stimulated by lipopolysaccharide and several other cytokines. In this study, the effect of sodium nitroprusside(SNP), a donor of nitric oxide($NO^-$), on osteoclast-like cell formation and on mature osteoclast function was examined. To determine the mechanism of the inhibitory effects of SNP decreased not only the basal $^{45}Ca$ release but also thee bone resorption induced by PTH and 1,25-dihydroxyvitamin $D_3\;(1,25[OH]_{2}D_3)$. The inhibitory effect of SNP on bone resorption induced by PTH appeared 2 dyas after treatment, whereas SNP effect on inhibiting bone resorption induced by $1,25[OH]_{2}D_3$ appeared at the thhird days. When chicken and rat osteeoclasts were cultured on dentin slices, treatment of $300{\mu}M$ SNP resulted in a significant decrease in dentin resorption by osteoclasts in terms of total resolution area and average individual area. We also examind the effect of SNP on formation of osteoclast-like cells that is TRAP-positive multinucleated cells from chicken and rat bone marrow cells in the presence or absence of $10^{-8}\;M\;1,25[OH]_{2}D_3$. The addition of $300{\mu}M$ SNP inhibiteed the formation of TRAP-positive multinucleated cells. The present data suggest that SNP, possibly as a $NO^-$ donor, inhibits the osteoclastic differentiation and osteoclastic activity.

• The korean journal of orthodontics
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• v.28 no.4 s.69
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• pp.627-640
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• 1998

[$TGF-{\beta}$] is a polypeptide with multiple physiological functions in regulation of cell-to-cell interaction and in growth and development. The active form of vitmain $D_3$, 1,25-dihydroxycholecalciferol $[1,25-(OH)_2D_3]$, is one of the most potent stimulators of osteoclastic acitivity. The purpose of this study was to evaluate the effect of Vitamin $D_3$ and/or $TGF-{\beta}$ on the periodontal ligament(PDL) cells. Human PDL cells were prepared from the first premolars extracted for the orthodontic treatment and were incubated in the environment of , $37^{\circ},\;5\%\;CO_2\;and\;95\%$ humidity. 10, 50 or 100ng/m1 of $1,25-(OH)_2D_3$ and 0.1, 1, 5 or 10ng/ml of $TGF-{\beta}$ were administered to the culture wells, separately or in combination. And the viability of PDL cells was evaluated by MTT assay The obtained results were as follows. 1. The viability of PDL cells in 10ng/ml of vitamin $D_3$ was not significantly differenent from that of the control group at 1, 2 and 3-day of cultivation, but it was significantly increased in 50ng/ml of Vitamin $D_3$ at 3-day and in 100ng/m1 of Vitamin $D_3$ at 2 and 3-day. 2. The viability of PDL cells in 0.1ng/ml of $TGF-{\beta}$ was not significantly differenent from that of the control group at 1, 2 and 3-day of cultivation, but it was significantly increased in 1 and 5ng/ml of $TGF-{\beta}$ at 3-day of cultivation, and in 10ng/ml of $TGF-{\beta}$ at 2 and 3-day of cultivation. 3. In case of admixture of 1ng/ml of $TGF-{\beta}$ and the various concentrations of vitamin $D_3$, the viability of PDL cells was significantly increased in the admixture of 100ng/ml of vitamin $D_3$ at 3-day of cultivation 4. In case of admixture of 5ng/ml of $TGF-{\beta}$ and the various concentrations of vitamin $D_3$, the viability of PDL cells began to be increased from 2-day of cultivation in the admixture of 10 50 and 100ng/ml of vitamin $D_3$, but it was not maintained at 3-day in the admixture of 10ng/m of vitamin $D_3$. 5. In case of admixture of 10ng/ml of $TGF-{\beta}$ and the various concentrations of vitamin $D_3$ the viability of PDL cells was significantly increased in the admixture of 50ng/ml of vitamin $D_3$ at 2 and 3-day of cultivation, and in the admixture of 100ng/ml at 1, 2 and 3-day.

• International Journal of Oral Biology
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• v.38 no.3
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• pp.127-134
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• 2013

Xylitol is a sugar alcohol with a variety of functions including bactericidal and anticariogenic effects. However, the cellular mechanisms underlying the role of xylitol in bone metabolism are not yet clarified. In our present study, we exploited the physiological role of xylitol on osteoclast differentiation in a co-culture system of osteoblastic and RAW 264.7 cells. Xylitol treatment of these co-cultures reduced the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells induced by 10 nM $1{\alpha},25(OH)_2D_3$ in a dose-dependent manner. A cell viability test revealed no marked cellular damage by up to 100 mM of xylitol. Exposure of osteoblastic cells to xylitol decreased RANKL, but not OPG, mRNA expression in the presence of $10^{-8}M$ $1{\alpha},25(OH)_2D_3$ in a dose-dependent manner. Furthermore, bone resorption activity, assessed on bone slices in the coculture system, was found to be dramatically decreased with increasing xylitol concentrations. RANKL and OPG proteins were assayed by ELISA and the soluble RANKL (sRANKL) concentration was decreased with an increased xylitol concentration. In contrast, OPG was unaltered by any xylitol concentration in this assay. These results indicate that xylitol inhibits $1{\alpha},25(OH)_2D_3$-induced osteoclastogenesis by reducing the sRANKL/OPG expression ratio in osteoblastic cells.