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The Effects of Mechanical Stress on Alkaline Phosphatase Activity of MC3T3-E1 Cells (기계적 자극이 MC3T3-EI 세포의 Alkaline Phosphatase Activity에 미치는 영향)

  • BAE, Sung-Min;KYUNG, Hee-Moon;SUNG, Jae-Hyun
    • The korean journal of orthodontics
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    • v.26 no.3
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    • pp.291-299
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    • 1996
  • Orthodontic force is a mechanical stress controlling both of tooth movement and skeletal growth. The mechanical stress stimulate bone cells that may exert some influence on bone remodeling. The purpose of this study was to evaluate the difference in cellular activity depending on mechanical stresses such as compressive and tensile force by determining the alkaline phosphatase(ALP) activity. A clonal osteogenic cell line MC3T3-E1 was seeded into a 24-well plate($2{\times}10^4/well$). At the confluent phase, a continuous compressive hydrostatic pressure($25g/cm^2$, $300g/cm^2$) and continuous tensile hydrostatic pressure($-25g/cm^2$, $-300g/cm^2$) were applied for 4, 6, 10, 14, 18, 20 days respectively by a diaphgragm pump. At the end of the stimulation period, cell layers were prepared for ALP activity assay. The ALP activity of the compressive group increased more than that of the tensile group at same force magnitude, whereas the cells responded to a similar pattern regardless of the type of mechanical stress The ALP activity of the compressive and tensile group turned into the level of the control group as the length of time increased. These results indicated that a mechanical stress may be more effective on cellular activity during active cellular proliferation and differentiation periods. The time to achieve maximum ALP activity was delayed as the mechanical stress increased in both the compressive and the tensile group. Accordingly, the magnitude of the stress rather than the type of mechanical stress may have more influence on cellular activity.

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Sequence characterization and polymorphism of melanocortin 1 receptor gene in some goat breeds with different coat color of Mongolia

  • Ganbold, Onolragchaa;Manjula, Prabuddha;Lee, Seung-Hwan;Paek, Woon Kee;Seo, Dongwon;Munkhbayar, Munkhbaatar;Lee, Jun Heon
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.7
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    • pp.939-948
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    • 2019
  • Objective: Extension and Agouti loci play a key role for proportions of eumelanin and pheomelanin in determining coat color in several species, including goat. Mongolian goats exhibit diverse types of coat color phenotypes. In this study, investigation of the melanocortin 1 receptor (MC1R) coding region in different coat colors in Mongolian goats was performed to ascertain the presence of the extension allele. Methods: A total of 105 goat samples representing three goat breeds were collected for this study from middle Mongolia. A 938 base pair (bp) long coding region of the MC1R gene was sequenced for three different breeds with different coat colors (Gobi Gurwan Saikhan: complete black, Zalaa Jinstiin Tsagaan: complete white, Mongolian native goat: admixture of different of coat colors). The genotypes of these goats were obtained from analyzing and comparing the sequencing results. Results: A total of seven haplotypes defined by five substitution were identified. The five single nucleotide polymorphisms included two synonymous mutations (c.183C>T and c.489G>A) and three missense (non-synonymous) mutations (c.676A>G, c.748T>G, and c.770T>A). Comparison of genotypes frequencies of two common missense mutions using chi-sqaure ($x^2$) test revealed significant differences between coat color groups (p<0.001). A logistic regression analysis additionally suggested highly significant association between genotypes and variation of black versus white uniform combination. Alternatively, most investigated goats (60.4%) belonged to H2 (TGAGT) haplotype. Conclusion: According to the findings obtained in this study on the investigated coat colors, mutations in MC1R gene may have the crucial role for determining eumelanin and pheomelanin phenotypes. Due to the complication of coat color phenotype, more detailed investigation needed.

The variation of insulin like growth factor 2 maker is associated with growth traits in Thai native (Kradon) pigs

  • Kessara Ampaporn;Rattikan Suwannasing;Pitchayanipa Phongphanich;Supanon Tunim;Monchai Duangjinda
    • Animal Bioscience
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    • v.36 no.9
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    • pp.1350-1356
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    • 2023
  • Objective: This study was conducted to investigate polymorphisms of the melanocortin-4 receptor (MC4R) and insulin like growth factor 2 (IGF2) genes and to evaluate the growth traits affected by such polymorphisms in Thai native (Kradon) pigs. Methods: Blood samples and productive data from 91 Kradon pigs were collected. DNA was extracted and quantified, the IGF2 and MC4R genes were amplified, and the polymerase chain reaction (PCR) produces were digested using the PCR-restriction fragment length polymorphism (PCR-RFLP) technique. Genotyping was performed, and the association between genotypes and growth traits on the birth and weaning weights were evaluated. Results: The IGF2 intron7 g.162G>C variations in Kradon pigs were found in three genotypes: i) GG, ii) GC, and iii) CC. The GG genotype frequency was the highest followed by the GC and CC genotypes. The frequencies of the G and C alleles were 0.703 and 0.297, respectively. The MC4R genotype was found in only one genotype (GG). The IGF2 gene pattern was not associated with birth weight traits, whereas the IGF2 gene pattern was related to the weaning weight trait in Kradon pigs. Pigs with the CC and GC genotypes had higher weaning weights than ones with the GG genotype (p<0.001). Conclusion: Thai native Kradon pigs with the CC and GC genotypes of the IGF2 gene have higher weaning weights than pigs with the GG genotype.

Effects of Scutellaria radix Extract on Osteoblast Differentiation and Osteoclast Formation (황금 추출물이 조골세포와 파골세포의 활성에 미치는 영향)

  • Shin, Jeong-Min;Park, Chan-Kyung;Shin, Eun-Ju;Jo, Tae-Hyung;Hwang, In-Kyeong
    • Korean Journal of Food Science and Technology
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    • v.40 no.6
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    • pp.674-679
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    • 2008
  • Scutellaria radix (SR) has been utilized as a traditional medicine for a variety of diseases including Rheumatoid arthritis and its major flavonoids - baicalein, baicalin, and wogonin - have been reported to exert beneficial health effects, including anti-bacterial, anti-viral, anti-inflammatory, and free-radical scavenging. However, the mechanisms underlying this effect remain poorly understood. The principal objective of this study was to determine the effect of SR on osteoblast and osteoclast cells. SR extract was prepared using 70% ethanol solvent. Osteoblastic MC3T3-E1 cells and osteoclast precursor Raw 264.7 macrophage cells were utilized. SR extract increased MC3T3-E1 cell proliferation and stimulated alkaline phosphatase activity dose-dependently, 152.0% of the control at concentration $1{\mu}g/mL$. Additionally, SR extract ($1{\mu}g/mL$) stimulated Bone nodule formation activity in MC3T3-E1 cells, approximately 223.3% of the control, 20 days after the exposure. In addition, SR extract significantly reduced the number of tartrate-resistant acid phosphatase-positive (TRAP+) multinucleated cells from Raw 264.7 cells. In conclusion, SR extract stimulates the proliferation and bioactivities of boneforming osteoblasts, and inhibits the activities of bone-resorbing osteoclasts to a certain degree.

Accelerating Effects of Quercetin on the $TNF-{\alpha}-Induced$ Apoptosis in MC3T3-E1 Osteoblastic Cells

  • Choi, Yong-Sung;Chung, Song-Woo;Jeon, Young-Mi;Kim, Jong-Ghee;Lee, Jeong-Chae
    • Natural Product Sciences
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    • v.11 no.3
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    • pp.139-144
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    • 2005
  • Bioflavone quercetin is believed to play an important role preventing bone loss by affecting osteoclastogenesis and regulating many systemic and local factors including hormones and cytokines. This study examined how quercetin acts on tumor necrosis factor-alpha ($TNF-{\alpha}$)-mediated apoptosis in MC3T3-E1 osteoblastic cells. Apoptosis assays revealed the dose-dependent acceleration of quercetin on $TNF-{\alpha}-induced$ apoptosis in MC3T3-E1 cells, which was demonstrated by the increased number of positively stained cells in the trypan blue staining and TUNEL assay, and the migration of many cells to the $sub-G_0/G_1$ phase in flow cytometric analysis. In particular, quercetin treatment alone increased the expression of p53 and p21 proteins in the cells. Consequently, this study showed that quercetin accelerates the $TNF-{\alpha}-induced$ apoptosis in MC3T3-E1 osteoblastic cells.

The effect of safflower seed fraction extract on periodontal ligament fibroblast and MC3T3-E1 cell in vitro (홍화씨 분획 추출물이 치주인대 섬유아세포와 MC3T3-E1 세포에 미치는 영향)

  • Huh, Ji-Sun;Kang, Jung-Hwa;Yoo, Yun-Jung;Kim, Chang-Sung;Cho, Kyoo-Sung;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.31 no.4
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    • pp.833-846
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    • 2001
  • Recently, use of natural medicine is getting more attention, and some of them are believed to be effective in the treatment of periodontitis. Among them, the seeds of safflower(Carthamus tinctrorius L.) have been proven to be effective through its use in bone diseases such as fracture and osteoporosis. During the last few years, studies using the seeds of safflower gown in Korea have been active, and it has been reported that safflower seed extract increase the proliferation and the alkaline phosphatase(ALP) activity of human periodontal ligament fibroblast(hPDLF), osteoblast, and that they promote the mineralization process. In animal studies, when safflower seed extract were administered orally new bone formation was promoted. Recently, in an effort to find out the most effective osteogenic components, among many components of the safflower seed, various safflower seed fraction extracts were obtained by multistep extraction of the safflower components using various solvents. Among these, saf-M-W fraction extracted by methanol and water was most effective in increasing osteogenic potential of osteoblasts. In this study, the effect of safflower seed fraction extract, saf-M-W, on the growth and differentiation of hPDLF and MC3T3-E1 cell was investigated. The toxicity of saf-M-W on both cells was measured using M'IT(3-(4,5dimethylthiazol-2-y1)-2,5-diphenyl tetrazolium bromide) test, and ALP activity was measured using the colorimetric assay of hPDLF. In addition, in MC3T3-El cells, the expression of ALP, bone sialoprotein(BSP) mRNA was observed using Northern blot, and the mineralized nodule formation Was observed using von Kossa stain and phase-contrast microscope. 1. In concentrations below $10{\mu}g/ml$, saf-M-W didn't show any toxicity on hPDLF and MC3T3-El cell. 2. The change in saf-M-W concentration had no effect on the ALP activity of hPDLF. 3. In MC3T-E1 cells, mRNA expressions of ALP and BSP were greater in the experimental group treated with $10{\mu}g/ml$ concentration of saf-M-W compared with the control group. 4. In MC3T3-El cells, abundance of mineralized nodules were formed in the experimental group treated with $10{\mu}g/ml$ Concentration of saf-M-W, while no mineralized nodule was formed in the control group. These results suggest that safflower seed fraction extract, saf-M-W. didn't show any toxicity on hPDLF and MC3T3-E1 cell at concentrations below $10{\mu}g/ml$ and effectively enhanced the differentiation and osteogenic potential of MC3T3-El cell.

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Antioxidative and Anti-aging Effects of Sancho (Zanthoxylum schinifolium) Extract in Rats Fed High Fat Diet

  • Jang Mi-Jin;Woo Mi-Hee;Rhee Soon-Jae;Cho Sung-Hee
    • Nutritional Sciences
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    • v.9 no.3
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    • pp.159-166
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    • 2006
  • This study was performed to investigate antioxidative and anti-aging action of extracts from Sancho (Zanthoxylum schinifolium) leaves. Two extracts were obtained by 80% methanol extraction followed by subsequent fractionation with methylene chloride (MC) and n-butanol (B) and fed at one or three levels to rats on normal level (5%) of fat (control) and high fat(20%) in diets. Male Sprague-Dawley rats weighing about 100 g were divided into ten groups such as control diet group(C), control diet+0.50%B group (CB), control diet+0.50%MC group (CMC), high-fat diet group (HF), high-fat diet+0.25%B group (HBL), high-fat diet+0.50%B group (HBM), high-fat diet+0.75%B group (HBH), high-fat diet+0.25%MC group (HMCL), high-fat diet+0.50%MC group (HMCM) and high-fat diet+ 0.75%MC group (HMCH) and fed each diet for four weeks. The effects of the extracts on antioxidant enzyme activities and indices of lipid peroxidation and aging were seen only in high fat diet groups. Hepatic superoxide dismutase and aryleaterase activities were not changed by Sancho extracts. But glutathione peroxidase, catalase and paraxonase activities were significantly restored by both MC and B at the level of 0.75% lipid peroxide which was increased by high fat diet was significantly reduced by B and MC at the level of 0.25% and over. Lipofuscin fluorescence and cabonyl value were increased by high fat diet were reduced by B and MC at the level of 0.5% and over. It is concluded that the Sancho extracts can be utilized as functional ingredients of health foods for reducing oxidative stress.

Anti-obesity effects of two herbal extracts in C57BL/6N mice fed high-fat diet

  • Hyuck Kim;Jihwan Lee;Seyeon Chang;Dongcheol Song;Jaewoo An;Sehyun Park;Kyeongho Jeon;Jungseok Choi;Jinho Cho
    • Korean Journal of Agricultural Science
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    • v.51 no.1
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    • pp.87-95
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    • 2024
  • The objective of this study was to investigate the anti-obesity effects of adding Momordica charantia (MC) and Chrysanthemum zawadskii var. latilobum (CZ) extracts to drinking water on obesity-induced mice. A total of 84 eight-week-old C57BL/6N male mice with an initial body weight (BW) of 28.11 ± 1.39 g were used in this study. All treatments were fed a high-fat diet for d 28. Mice were randomly divided into seven drinking treatments (six replicate each treatment) based on the initial BW. Treatments are as follows: control (CON), normal tap water, MC 1, CON with 1% MC aqueous extract, MC 2, CON with 2% MC aqueous extract, CZ 1, CON with 1% CZ aqueous extract, CZ 2, CON with CZ aqueous extract (2%), MCZ 1, CON with 1% MC aqueous extract and 1% CZ aqueous extract, MCZ 2, CON with 2% MC aqueous extract and 2% CZ aqueous extract. During the entire period, the CZ 1, MCZ 1, and MCZ 2 significantly decreased (p < 0.05) gain to feed than CON. The CON significantly higher (p < 0.05) water intake than other treatments on d 0 to 14. The MCZ 1 significantly decreased (p < 0.05) relative (ratio of absolute organ weight to BW) organ weights, including retroperitoneal white adipose tissue (RWAT) weight and inguinal white adipose tissue (IWAT) weight, compared to CON. In conclusion, our study suggests that there was no significant difference in the anti-obesity effects between MC and CZ, and MCZ 1 has synergistic effects by regulating adipose tissue.

A Study on the Changes of Coat Color-Related Genes according to Generational Changes in Jeju Horses (제주마 집단의 세대 경과에 따른 모색 유전자 변화 연구)

  • Kim, Nam-Young;Chae, Hyun-Seok;Baek, Kwang-Soo;Cho, In-Chul;Jung, Young-Hun;Woo, Jae-Hoon;Park, Seol-Hwa;Kim, Ji-Hyang;Lee, Sung-Soo;Yang, Young-Hoon
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.183-188
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    • 2015
  • This study analyzed the coat color-related genes of MC1R, ASIP, ECA3-inversion, and STX17 of 1,462 Jeju horses administered by the Jeju Special Self-Governing Province. This was done to investigate the distributional characteristics of coat color-related genes in the Jeju horse group and the changes of its coat color-related genes by generation. The genotype frequency of the MC1R gene of $E^+/E^+$ and $E^+/E^e$ related to black coat color was 0.122 and 0.447, respectively, while $E^e/E^e$ of the chestnut genotype was 0.429. The genotype frequency of the ASIP gene of $A^A/A^A$, $A^A/A^a$, and $A^a/A^a$ was 0.46, 0.448, and 0.091, respectively, where the genotype frequency of $A^a/A^a$ turned out to be relatively low. The To/To and +/To genotype that manifests the Tobiano shape was 0.001 and 0.119, respectively, with the share of Tobiano shape around 12%. The genotype frequency of G/G and G/g of STX17 related to grey coat color was 0.002 and 0.680, respectively, with the share of grey horses among the Jeju horse group at 68.2%. As for the change of coat color genes by generation, no large changes were observed in the MC1R and ASIP genes. In ECA3-inversion, the To allele that manifests Tobiano significantly decreased following the generational change (p<0.05), while the STX17 G allele related to grey coat color significantly increased following the generational change (p<0.05). It will be necessary to examine the coat color genes when selecting breeding horses so that the diversity of coat colors among the Jeju horse group can be maintained.

Degradation of Microcystin-LR, Taste and Odor, and Natural Organic Matter by UV-LED Based Advanced Oxidation Processes in Synthetic and Natural Water Source (UV-LED기반 고도산화공정을 이용한 수중 마이크로시스틴-LR, 이취미 물질, 자연유기물 분해)

  • Yang, Boram;Park, Jeong-Ann;Nam, Hye-Lim;Jung, Sung-Mok;Choi, Jae-Woo;Park, Hee-Deung;Lee, Sang-Hyup
    • Journal of Korean Society of Environmental Engineers
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    • v.39 no.5
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    • pp.246-254
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    • 2017
  • Microcystin-LR (MC-LR) is one of most abundant microcystins, and is derived from blue-green algae bloom. Advanced oxidation processes (AOPs) are effective process when high concentrations of MC-LR are released into a drinking water treatment system from surface water. In particular, UV-based AOPs such as UV, $UV/H_2O_2$, $UV/O_3$ and $UV/TiO_2$ have been studied for the removal of MC-LR. In this study, UV-LED was applied for the degradation of MC-LR because UV lamps have demonstrated some weaknesses, such as frequent replacements; that generate mercury waste and high heat loss. Degradation efficiencies of the MC-LR (initial conc. = $100{\mu}g/L$) were 30% and 95.9% using LED-L (280 nm, $0.024mW/cm^2$) and LED-H (280 nm, $2.18mW/cm^2$), respectively. Aromatic compounds of natural organic matter changed to aliphatic compounds under the LED-H irradiation by LC-OCD analysis. For application to raw water, the Nak-dong River was sampled during summer when blue-green algae were heavy bloom in 2016. The concentration of extracellular and total MC-LR, geosmin and 2-MIB slightly decreased by increasing the LED-L irradiation; however, the removal of MC-LR by UV-LED (${\lambda}=280nm$) was insufficient. Thus, advanced UV-LED technology or the addition of oxidants with UV-LED is required to obtain better degradation efficiency of MC-LR.