• Journal of Life Science
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• v.27 no.9
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• pp.1031-1039
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• 2017

The aim of this study was to investigate the potential effects of extracts from silkworm Bombyx mori L. fermented with Bacillus subtilis KACC 91157 at levels of 5%(v/w) and 10%(v/w) in Sprague-Dawley rats intoxicated with 1%(w/w) orotic acid (OA) for 10 days. The rats were divided into a normal group (N), a control group (C: OA), and treatment groups (SP10: OA + 10% extracts from B. mori L.; BSP5: OA + 5% extracts from B. mori L. fermented with B. subtilis KACC 91157; BSP10: OA + 10% extracts from B. mori L. fermented with B. subtilis KACC 91157). Serum activities of aspartate aminotransferase (AST), alanine transferase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) increased following OA feeding, but the rise was slightly reduced by administration of BSP10. The total lipid, free fatty acid, phospholipid, total cholesterol, and triglyceride contents in serum were significantly lower in the OA treatment groups than in the N group. However, the contents slightly increased following the administration of BSP10. Glutathione concentrations in liver and serum were reduced in the OA-induced fatty liver, but they increased following the administration of BSP10. Hepatocytes in the OA-induced fatty liver contained numerous large droplets. However, SP10, BSP5, and BSP10 feeding prevented OA-induced lipid droplet accumulation in hepatocytes. Accordingly, extracts from silkworm powder fermented with B. subtilis could be an ideal material as a dietary supplement in healthy functional foods to improve the effects of fatty liver.

• Polymer(Korea)
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• v.39 no.3
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• pp.418-425
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• 2015

Polycaprolactone (PCL) and biphasic calcium phosphate (BCP) have been considered as useful materials for orthopedic devices and osseous implants because of their biocompatibility and bone-forming activity. However, PCL-based scaffolds have hydrophobic surfaces reducing initial cell adhesion or proliferation. To overcome the limitation, we fabricated surface-modified PCL/BCP nanofibers using gamma-irradiation for bone tissue engineering. PCL/BCP nanofibers were prepared by electrospinning and then we supplemented hydrophilicity by introducing acrylic acid (AAc) through gamma-irradiation. We confirmed the surface of nanofibers by SEM, and then the initial viability of MG63 was significantly increased on the AAc grafted nanofibers, and alkaline phosphatase activity($1.239{\pm}0.226nmole/{\mu}g/min$) improved on the modified nanofibers than that on the non-modified nanofibers($0.590{\pm}0.286nmole/{\mu}g/min$). Therefore, AAc-grafted nanofibers may be a good tool for bone tissue engineering applications.

• Journal of Oral Medicine and Pain
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• v.26 no.4
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• pp.319-329
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• 2001

Endothelin-1 (ET-1) is a recently discovered potent vasoconstrictive peptide. It was first identified in vascular endothelial cells. ET-1 is a 21-amino acid peptide and elicits systemic effects such as stimulation of the production of atrial natriuretic peptide and release of aldosterone and corticosterone. In this study, to examine the role of ET-1 in the bone metabolism, effect of ET-1 on the proliferation and activity of osteoblastic cells was studied using HOS cells as osteoblast model. ET-1 dose-dependently increased the cell proliferation as determined by cell counting and MTT reduction assay after 48hr treatment. Alkaline phosphatase activity was inhibited by ET-1 and showed significant inhibition by 50 and 100 nM ET-1. ET-1 increased NBT reduction by HOS cells dose-dependently showing that ET-1 may increase the superoxide production by osteoblasts. Nitrite concentration in the media of HOS cell culture without cytokine stimulation was negligible and unaffected by ET-1 after 48hr treatment. Finally, after collection and concentration of conditioned media, gelatinase activity produced by HOS cells was determined by zymography. HOS cells can produce and secrete the gelatinase (gelatinase A type as determined by molecular weight of about 65,000) into culture media, however, ET-1 had no effect on the gelatinase activity. These findings suggest that ET-1 may have diverse effects on the proliferation and differentiation of osteoblasts, therefore, it may play an important role in bone metabolism.

• Nutrition Research and Practice
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• v.12 no.4
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• pp.275-282
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• 2018

BACKGROUND/OBJECTIVE: There is intense interest in soy isoflavone as a hormone replacement therapy for the prevention of postmenopausal osteoporosis. A new kind of isoflavone-enriched whole soy milk powder (I-WSM) containing more isoflavones than conventional whole soy milk powder was recently developed. The aim of this study was to investigate the effects of I-WSM on bone metabolism in ovariectomized mice. MATERIALS/METHODS: Sixty female ICR mice individually underwent ovariectomy (OVX) or a sham operation, and were randomized into six groups of 10 animals each as follows: Sham, OVX, OVX with 2% I-WSM diet, OVX with 10% I-WSM diet, OVX with 20% I-WSM diet, and OVX with 20% WSM diet. After an 8-week treatment period, bone mineral density (BMD), calcium, alkaline phosphatase (ALP), tartrate-resistant acid phosphatase (TRAP) 5b, osteocalcin (OC), procollagen 1 N-terminal propeptide (P1NP), and osteoprotegenin (OPG) were analyzed. RESULTS: BMD was significantly lower in the OVX group compared to the Sham group but was significantly higher in OVX + 10% I-WSM and OVX + 20% I-WSM groups compared to the OVX group (P < 0.05). Serum calcium concentration significantly increased in the OVX + 10% and 20% I-WSM groups. Serum ALP levels were significantly lower in the OVX + 10% and 20% I-WSM groups compared to the other experimental groups (P < 0.05). OC was significantly reduced in the OVX group compared to the Sham group (P < 0.05), but a dose-dependent increase was observed in the OVX groups supplemented with I-WSM. P1NP and OPG levels were significantly reduced, while TRAP 5b level was significantly elevated in the OVX group compared with the Sham group, which was not affected by I-WSM (P < 0.05). CONCLUSIONS: This study suggests that I-WSM supplementation in OVX mice has the effect of preventing BMD reduction and promoting bone formation. Therefore, I-WSM can be used as an effective alternative to postmenopausal osteoporosis prevention.

• The Journal of Korean Obstetrics and Gynecology
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• v.25 no.3
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• pp.16-26
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• 2012

Objectives: This study was performed to evaluate effects of Cordyceps militaris(CM) on osteoclast differentiation and its related gene expression. Methods: We used mouse myeloid cells RAW 264.7 stimulated by receptor activator of nuclear factor kappa-B ligand(RANKL) to induce osteoclast differentiation. There are four groups of which RAW 264.7 cells are not stimulated by RANKL (Normal), stimulated by RANKL without CM(Control), stimulated by RANKL with 0.1 ${\mu}g/ml$ of CM(CM 0.1), stimulated by RANKL with 1 ${\mu}g/ml$ of CM(CM 1). Osteoclastogenesis was measured by counting Tartrate-resistant acid phosphatase-positive multinucleated cells [TRAP(+) MNC]. RT-PCR was performed to evaluate the inhibitory effect of CM on gene expression(TRAP, AKT1, JNK1, NFATc1, c-Fos, MITF). Results: 1. CM decreased the number of TRAP(+) osteoclast in RANKL-stimulated RAW 264.7 cell at the concentration of 0.1 ${\mu}g/ml$ and 1 ${\mu}g/ml$. 2. CM decreased the expression of TRAP in osteoclast at the concentration of 1 ${\mu}g/ml$. 3. CM decreased the expression of AKT1, JNK1 in osteoclast at the concentration of 1 ${\mu}g/ml$. 4. CM didn't affect the expression of NFATc1, c-Fos, MITF in osteoclast. Conclusions: Cordyceps militaris has inhibitory effects on osteoclast differentiation and its related gene expression. These results suggest that Cordyceps militaris has a potential as a treatment of osteoporosis.

• BMB Reports
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• v.52 no.11
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• pp.659-664
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• 2019

Vav1 is a Rho/Rac guanine nucleotide exchange factor primarily expressed in hematopoietic cells. In this study, we investigated the potential role of Vav1 in osteoclast (OC) differentiation by comparing the ability of bone marrow mononuclear cells (BMMCs) obtained from Vav1-deficient ($Vav1^{-/-}$) and wild-type (WT) mice to differentiate into mature OCs upon stimulation with macrophage colony stimulating factor and receptor activator of nuclear kappa B ligand in vitro. Our results suggested that Vav1 deficiency promoted the differentiation of BMMCs into OCs, as indicated by the increased expression of tartrate-resistant acid phosphatase, cathepsin K, and calcitonin receptor. Therefore, Vav1 may play a negative role in OC differentiation. This hypothesis was supported by the observation of more OCs in the femurs of $Vav1^{-/-}$ mice than in WT mice. Furthermore, the bone status of $Vav1^{-/-}$ mice was analyzed in situ and the femurs of $Vav1^{-/-}$ mice appeared abnormal, with poor bone density and fewer number of trabeculae. In addition, Vav1-deficient OCs showed stronger adhesion to vitronectin, an ${\alpha}_v{\beta}_3$ integrin ligand important in bone resorption. Thus, Vav1 may inhibit OC differentiation and protect against bone resorption.

• Korean Journal of Soil Science and Fertilizer
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• v.38 no.1
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• pp.15-24
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• 2005

Enterobacter intermedium oxidizes glucose to gluconic acid and sequentially converts gluconic acid to 2-ketogluconic acid (2-KGA) under aerobic condition. Shaking incubation of E. intermedium in a broth medium containing 22.5 g glucose, 8.2 g gluconic acid and 40 g rock phosphate per liter resulted in $1028mg\;L^{-1}$ soluble phosphate. The culture broth was used as phosphate bio-fertilizer (PBF) in this experiment. To evaluate PBF produced by E. intermedium on lettuce growth, five treatments (PBF1/3, PBF2/3, PBF3/3, BP, and MF) were used. In MF and BP treatments, $P_2O_5$ 5.9 kg of mineral fertilizer per 10a was added, while in PBF1/3, PBF2/3, and PBF3/3 treatments, culture broth containing one third, two third, and same amount of soluble $P_2O_5$ 5.9 kg per 10a was applied, respectively. At 20, 35, and 50 days after transplanting of lettuce, plant growth components, biomass, enzyme activities and soil chemical properties were analyzed. Dehydrogenase activity and available phosphate concentration of rhizosphere in phosphate bio fertilizer treatments (PBF1/3, PBF2/3, PBF3/3) were generally higher compared to MF and BP treatments. Soil biomass in PBF3/3 treatment was significantly higher than MF and BP treatments at early growth stage. However, there was no significant difference among all treatments with time. Plant fresh weights in PBF1/3, PBF2/3, and MF treatments were better than those in BP and PBF3/3 treatments. However, in PBF2/3 treatment the highest fresh weight was discovered where alkaline phosphatase activity was generally higher than other treatments at 35 and 50 days. Enhancement of lettuce growth at 35 and 50 days in PBF2/3 treatment was associated with greater phosphate uptake in lettuce tissue. As regarding all results, PBF showed better lettuce growth compared to mineral phosphate fertilizer where PBF2/3 treatment resulted in increase of lettuce fresh weight by 23% and phosphate uptake by 50%.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.577-598
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• 1996

This study was conducted to identify the effects of caffeine or combinations of caffeine and iron or vitamin E on the lipid and protein components or blood chemistry levels of the serum as well as the total homogenate, mitochondrial and microsomal fraction of the rat(Sprague-Dawley, female) liver. Chronic test were conducted to determine those effects. The chronic test was conducted by dividing rats into 5 groups according to the type of drugs and dosages administrated as follows; the control(group A), and group B was given 25mg/kg caffeine orally once daily for 30 days, group C was given 50mg/kg caffeine orally once daily for 30 days, group D was given 25mg/kg caffeine and orally ferric chloride once daily for 30 days and group E was given 25mg/kg caffeine and 25mg/kg vitamin E once daily for 30 days. The concentrations of glucose, urea nitrogen, uric acid, creatinine, total protein, albumin, A/G ratio, triglyceride, total cholesterol, HDL-cholesterol, free fatty acid, phospholipid as well as the activities of alanine aminotransferase(ALT), aspartate aminotransferase(AST) and alkaline phosphatase(ALP) were measured in the serum of each experimental groups. The concentrations of the carbonyl group and malondiaidehyde(MDA) and the patterns of the SDS-PAGE(Sodium Dodecyl Sulfate - Polyacrylamide Gel Electrophoresis) and fatty acid compositions in free fatty acids and phospholipids were analyzed to determine the oxidative damages and metabolic changes on the lipid and protein components in the serum, and total homogenate, mitochondrial and microsomal fractions of the rat liver. The results obtained from this study were summarized as follows; 1. Body weights of groups B, C, D and E were significantly decreased(p < 0.01) in comparison with that of the control in the chronic test. 2. The concentrations of serum glucose in groups B(124.5mg/dl), C(130.1mg/dl), D(122.1mg/dl), E(119.3mg/dl) were significantly higher(p < 0.01) in comparison to that of the control(101.5mg/dl). But, there were no significant differences in the concentrations of urea nitrogen, uric acid, creatinine, total protein, albumin and A/G ratio in comparison to that of the control. 3. The concentrations of total cholesterol and HDL-cholesterol in serum of groups B(69.6, 53.4mg/dl), C(73.0, 56.3mg/dl), D(68.9, 51.1mg/dl) and E(68.2, 51.3mg/dl) were significantly higher(p < 0.01) in comparison to that of the control(52.6, 38.8mg/dl). On the other hand, the concentrations of triglyceride in serum of groups B(45.0mg/dl), C(40.4mg/dl), D(33.8mg/dl) and E(47.2mg/dl) were significantly lower(p < 0.01) in comparison to that of the control(66.2mg/dl). There were no significant differences in the activities of ALT, AST and ALP in comparison to that of the control. 4. The concentrations of free fatty acid and phospholipid in serum of groups B(45.7, 154.4mg/dl), C(50.0, 167.2mg/dl), D(52.5, 148.4mg/dl) and E(41.1, 159.2mg/dl) were higher(p < 0.01) in comparison to that of the control(35.2, 125.3mg/dl). And the concentrations of the carbonyl group and malondialdehyde in serum of group D(1.82, 0.52nM/mg protein) were significantly higher(p < 0.01) in comparison to the control(1.53nM/mg protein). 5. The concentrations of carbonyl group in total homogenate, mitochondrial and microsomal fraction of group D(1.45, 0.94, 1.67nM/mg protein) were significantly higher (p < 0.01) in comparison to the control(1.16, 0.66, 1.27nM/mg protein). And the concentrations of malondialdehyde in the total homogenate, mitochondrial and microsomal fraction of group D(6.70, 6.10, 1.36nM/mg protein) were significantly higher(p < 0.01) in comparison to the control(5.17, 3.64, 0.68nM/mg protein). 6. As the analytical results of the fatty acid compositions of free fatty acid in serum, the proportions of stearic acid and arachidonic acid of groups B(16.52, 12.62%), C(17.52, 15.18%), D(19.73, 13.47%) and E(17.62, 13.28%) were significantly higher(p < 0.01) in comparison to the control(14.75, 7.88%), but the proportions of oleic acid and linoleic acid of groups B(12.97, 32.59%), C(10.88, 31.23%), D(12.37, 30.66%) and E(11.95, 32.41%) were significantly lower(p < 0.01) in comparison to the control(16.44, 35.12%). Otherwise, as the results of the fatty acid compositions of phospholipid in serum, the proportions of stearic acid and arachidonic acid of groups B(39.37, 16.39%), C(40.63, 17.83%), D(42.73, 15.39%) and E(39.16, 15.70%) were significantly higher(p < 0.01) in comparison to the control(37.74, 14.24%), but the proportions of oleic acid and linoleic acid of groups B(4.03, 14.38%), C(3.54, 12.38%), D(4.52, 11.68%) and E(4.29, 13.64%) were significantly lower(p < 0.01) in comparison to the control(5.53, 16.14%). 7. As the analytical results of the fatty acid compositions of free fatty acid in total homogenate, mitochondrial and microsomal fraction of liver, the proportions of oleic acid of groups B(7.8**, 8.73**, 6.88%) and C(6.89**, 7.75**, 6.58%) were lower(**:p < 0.01) in comparison to the control(8.67, 10.08, 7.81%), but the proportions of arachidonic acid of group C(22.62, 19.79, 23.71%) were significantly higher(p < 0.01) in comparison to the control(20.93, 18.47, 22.24%). And the proportions of palmitic acid of group D(25.95**, 26.16, 26.34**%) were significantly higher(**:p < 0.01) in comparison to the control(24.43, 25.42, 23.34%). In addition, the proportions of linoleic acid of group D(23.43, 25.02, 23.95%) were also significantly higher(p < 0.01) in comparison to the control(22.17, 23.75, 21.26%). The proportions of stearic acid of group D(19.87, 19.76**%) in mitochondrial and microsomal fraction were lower(**:p < 0.01) in comparison to the control(21.01, 24.18%), and the proportions of stearic acid of group E(16.71*, 19.65**%) in mitochondrial and microsomal fraction were significantly lower(**:p < 0.01, *:p < 0.05) in comparison to the control(21.01, 24.18%), and the proportions of linoleic acid of group E(25.04, 29.20, 26.48%) in total homogenate, mitochondria and microsome were significantly higher(p < 0.01) in comparison to the control(22.17, 23.75, 21.26%). 8. As the results of the fatty acid compositions of phospholipid in total homogenate, mitochondrial and microsomal fraction of liver, the proportions of palmitic acid of group D(17.58**, 18.78*, 18.23%**) were significantly higher(**:p < 0.01, *:p < 0.05) in comparison to the control(16.28, 17.22, 16.38%), and the proportions of stearic acid of group D(36.41, 37.23, 39.53%) were also significantly higher(p < 0.01) in comparison to the control(34.18, 34.16, 36.04%). But the proportions of oleic acid(3.41*, 3.11**, 3.12**%) and linoleic acid (18.03**, 15.79**, 14.74**%) of group D were significantly lower(**:p < 0.01, *:p < 0.05) in comparison to the control(oleic : 3.63, 3.72, 3.79%, linoleic : 20.03, 18.71, 18.48%). 9. In order to determine the oxidative damages to the protein in serum, mitochondrial and microsomal fraction of the rat liver, the patterns of the SDS-PAGE were identified, but the results of SDS-PAGE were not significantly different between the control and experimental groups.

• BMB Reports
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• v.51 no.7
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• pp.362-367
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• 2018

A major feature of type 1 diabetes mellitus (T1DM) is hyperglycemia and dysfunction of pancreatic ${\beta}$-cells. In a previous study, we have shown that Tat-DJ-1 protein inhibits pancreatic RINm5F ${\beta}$-cell death caused by oxidative stress. In this study, we examined effects of Tat-DJ-1 protein on streptozotocin (STZ)-induced diabetic mice. Wild type (WT) Tat-DJ-1 protein transduced into pancreas where it markedly inhibited pancreatic ${\beta}$-cell destruction and regulated levels of serum parameters including insulin, alkaline phosphatase (ALP), and free fatty acid (FFA) secretion. In addition, transduced WT Tat-DJ-1 protein significantly inhibited the activation of $NF-{\kappa}B$ and MAPK (ERK and p38) expression as well as expression of COX-2 and iNOS in STZ exposed pancreas. In contrast, treatment with C106A mutant Tat-DJ-1 protein showed no protective effects. Collectively, our results indicate that WT Tat-DJ-1 protein can significantly ameliorate pancreatic tissues in STZ-induced diabetes in mice.

• The Korean Journal of Physiology and Pharmacology
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• v.14 no.3
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• pp.169-176
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• 2010

The hyperosmotic stimulus is regarded as a mechanical factor for bone remodeling. However, whether the hyperosmotic stimulus affects $1{\alpha}$, 25-dihydroxyvitamin $D_3$ ($1{\alpha},25(OH)_2D_3$)-induced osteoclastogenesis is not clear. In the present study, the effect of the hyperosmotic stimulus on $1{\alpha},25(OH)_2D_3$-induced osteoclastogenesis was investigated in an osteoblast-preosteoclast co-culture system. Serial doses of sucrose were applied as a mechanical force. These hyperosmotic stimuli significantly evoked a reduced number of $1{\alpha},25(OH)_2D_3$-induced tartrate-resistant acid phosphatase-positive multinucleated cells and $1{\alpha},25(OH)_2D_3$-induced bone-resorbing pit area in a co-culture system. In osteoblastic cells, receptor activator of nuclear factor ${\kappa}B$ ligand (RANKL) and Runx2 expressions were down-regulated in response to $1{\alpha},25(OH)_2D_3$. Knockdown of Runx2 inhibited $1{\alpha},25(OH)_2D_3$-induced RANKL expression in osteoblastic cells. Finally, the hyperosmotic stimulus induced the overexpression of TonEBP in osteoblastic cells. These results suggest that hyperosmolarity leads to the down-regulation of $1{\alpha},25(OH)_2D_3$-induced osteoclastogenesis, suppressing Runx2 and RANKL expression due to the TonEBP overexpression in osteoblastic cells.