• International Journal of Industrial Entomology and Biomaterials
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• 제27권1호
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• pp.159-165
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• 2013

Bone morphogenetic proteins (BMPs) belong to the transforming growth factor (TGF-${\beta}$) superfamily and are involved in osteoblastic differentiation. The largest TGF-${\beta}$ superfamily subgroup shares genetic homology with human BMPs (hBMPs) and silkworm decapentaplegic (dpp). In addition, hBMPs are functionally interchangeable with Drosophila dpp. Bombyx mori dpp may induce bone formation in mammalian cells. To test this hypothesis, we synthesized the 1,285-base pairs cDNA of full-length B. mori dpp using total RNAs obtained from the fat body of 3-day-old of the $5^{th}$ instar larvae and cloned the cDNA into the pCEP4 mammalian expression vector. Next, B. mori dpp was expressed in C3H10T1/2 cells. The target cells transfected with the pCEP4-Bm dpp plasmid showed biological functions similar to those of osteogenic differentiation induction growth factors such as hBMPs. We determined the relative mRNA expression rates of Runt-related transcription factor 2 (RUNX2), osterix, osteocalcin, and alkaline phosphatase (ALP) to validate the osteoblast-specific differentiation effects of B. mori dpp by performing quantitative real-time RT-PCR. Interestingly, mRNA expression levels of the 3 marker genes except RUNX2, in cells expressing B. mori dpp were much higher than those in control cells and C3H10T1/2 cells transfected with pCEP4. These results suggested that B. mori dpp signaling regulates osterix expression during osteogenic differentiation via RUNX2-independent mechanisms.

• 한국임상수의학회지
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• 제15권2호
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• pp.442-449
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• 1998

Freeze-dried cortical bones of the goat were transplanted to the experimental fibular defect of 10 dogs for valuating the possibility of xenogeneic bone implantation and the specificity of BM(Bone Morphogenetic Protein). The . freeze-dried cortical bone eliminated antigens and defatted with chloroform and methanol were freeze-dried at $-80{\circ}C$ for preservation of BMP and then sterilized with 50 gas and storaged in room temperature. Ten freeze-dried cortical implants of the goat were transplanted in experimentally defected regions of bilateral fibula of 5 dogs in clinically normal. The transplanted region had been radiographed for observing state of bone union and BALPOone Alkaline Phosphatase) in the serum of the host was measured for valuating activity of oteoblast per 2 week-interval after transplant procedures. New bone formation had been observed early in one of ten regions around implants about the same time as autoimplant regions. It was incorporated with its host bone during 4-12 weeks after transplantation. In another 2 cases of 2 dogs, new bone formation and absorption of implant had been observed from 4 weeks but they were not incorporated completely until 20 weeks. The rest of the freeze-dried bone implants, 7 cases of 4 dogs had not been observed new bone formation nor absorption of implants. The freeze-drying method for implants means to not influence bone incorporation. Although less of union percentages the union form of this experiment were similar to alloimplantation and it may mean to block immunity reaction that disturbs the bone induction by BMP. It demonsknted that the possibility of the xenogenous bone implantation is recognized by reason of the low specificity of BMP between goat and dog.

• Journal of Periodontal and Implant Science
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• 제41권4호
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• pp.167-175
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• 2011

Purpose: Periodontal ligament (PDL) cell differentiation into osteoblasts is important in bone formation. Bone formation is a complex biological process and involves several tightly regulated gene expression patterns of bone-related proteins. The expression patterns of bone related proteins are regulated in a temporal manner both in vivo and in vitro. The aim of this study was to observe the gene expression profile in PDL cell proliferation, differentiation, and mineralization in vitro. Methods: PDL cells were grown until confluence, which were then designated as day 0, and nodule formation was induced by the addition of 50 ${\mu}g$/mL ascorbic acid, 10 mM ${\beta}$-glycerophosphate, and 100 nM dexamethasone to the medium. The dishes were stained with Alizarin Red S on days 1, 7, 14, and 21. Real-time polymerase chain reaction was performed for the detection of various genes on days 0, 1, 7, 14, and 21. Results: On day 0 with a confluent monolayer, in the active proliferative stage, c-myc gene expression was observed at its maximal level. On day 7 with a multilayer, alkaline phosphatase, bone morphogenetic protein (BMP)-2, and BMP-4 gene expression had increased and this was followed by maximal expression of osteocalcin on day 14 with the initiation of nodule mineralization. In relationship to apoptosis, c-fos gene expression peaked on day 21 and was characterized by the post-mineralization stage. Here, various genes were regulated in a temporal manner during PDL fibroblast proliferation, extracellular matrix maturation, and mineralization. The gene expression pattern was similar. Conclusions: We can speculate that the gene expression pattern occurs during PDL cell proliferation, differentiation, and mineralization. On the basis of these results, it might be possible to understand the various factors that influence PDL cell proliferation, extracellular matrix maturation, and mineralization with regard to gene expression patterns.

• Genomics & Informatics
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• 제2권2호
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• pp.75-80
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• 2004

In a variety of animal species, the perinatal exposure of experimental animals to the 2,3,7,8-tetrachlorodibenzo­p-dioxin (TCDD) leads to the immune dysfunction, which is more severe and persistent than that caused by adult exposure. We report here the changes of gene expression and the identification of the marker-genes representing the dioxin exposure. The expressions of the transcripts were analyzed using the 11 K oligonucleotide­microarray from the bone marrow cells of male C57BL/6J mice after an intraperitoneal injection of $1{\mu}g$ TCDD/kg body weight at various time intervals: gestational 6.5 day(G6.5), 13.5 day(G13.5), 18.5 day(G18.5), and postnatal 3 (P3W)and 6 week (P6W). The type of self-organizing maps(SOM) representing the specific exposure dioxin could be identified as follows; G6.5D(C14), G13.5D(C0, C5, C10, C18), G18.5D(7): P3W(C2, C21), and P6W(C4, C15, C20). The candidate marker-genes were restricted to the transcripts, which could be consistently expressed greater than $\pm$2-fold in three experiments. The resulting candidates were 85 genes, the characteristics of that were involved in cell physiology and cell functions such as cell proliferation and immune function. We identified the biomarker-genes for dioxin exposure: smc -like 2 from SOM C14 for the dioxin exposure at G6.5D, focal adhesion kinase and 6 other genes from C0, and protein tyrosine phosphatase 4a2 and 3 other genes from C5 for G13.5D, platelet factor 4 from C7 for G18.5D, fos from C2 for P3W.

• The Korean Journal of Physiology and Pharmacology
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• 제10권5호
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• pp.243-249
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• 2006

The effect of genistein, widely used as a specific tyrosine kinase inhibitor, on rat brain Kv1.5 channels which were stably expressed in Chinese hamster ovary cells was investigated using the whole-cell patch-clamp technique. Genistein inhibited Kv1.5 currents at +50 mV in a concentration-dependent manner, with an $IC_{50}$ of $54.7{\pm}8.2\;{\mu}M$ and a Hill coefficient of $1.1{\pm}0.2$. Pretreatment of Kv1.5 with protein tyrosine kinase inhibitors ($10\;{\mu}M$ lavendustin A and $100\;{\mu}M$ AG1296) and a tyrosine phosphatase inhibitor ($500\;{\mu}M$ sodium orthovanadate) did not block the inhibitory effect of genistein. The inhibition of Kv1.5 by genistein showed voltage-independence over the full activation voltage range positive to 0 mV. The activation (at +50 mV) kinetics was significantly delayed by genistein: time constant for an activation of $1.4{\pm}0.2$ msec under control conditions and $10.0{\pm}1.5$ msec in the presence of $60\;{\mu}M$ genistein. Genistein also slowed the deactivation of the tail currents, resulting in a crossover phenomenon: a time constant of $11.4{\pm}1.3$ msec and $40.0{\pm}4.2$ msec under control conditions and in the presence of $60\;{\mu}M$ genistein, respectively. Inhibition was reversed by the application of repetitive depolarizing pulses, especially during the early part of the activating pulse. These results suggest that genistein directly inhibits Kv1.5 channels, independent of phosphotyrosine-signaling pathway.

• 한국토양비료학회지
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• 제22권2호
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• pp.93-99
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• 1989

보릿짚시용(施用)이 수도재배(水稻栽培) 논 토양(土壤)에 미치는 영향(影響)을 규명하기 위하여 투수(透水)를 실시하여 토양중(土壤中) 질소고정(窒素固定)에 관여하는 미생물(微生物), Acetylene 환원력(還元力), 효소활성(酵素活性), 당(糖)을 조사분석(調査分析)한 결과(結果)를 요약(要約)하면 다음과 같다. 1. 질소고정(窒素固定) 미생물중(微生物中) Azotobacter는 경시적(經時的)으로 증가(增加)하였고 재배구(栽培區)에서 보릿짚시용(施用)에 의해 $17.61{\times}10^4$ 으로 증가(增加)하였으며 Clostridia는 재배구(栽培區)에서 보릿짚시용(施用)에 의해 분열기에 $34.56{\times}10^4$ 으로 증가(增加)하였다가 그 후(後)에는 $0.3{\times}10^4$ 으로 감소(減少)하였다. Blue-green algae는 출수기에 $18.0{\times}10^3$ 으로 증가(增加)하였다가 그 후(後)에는 $0.46{\times}10^4$ 으로 감소(減少)하였는데 보릿짚시용(施用)에 의해 증가(增加)하는 경향(傾向)이었다. 2. Acetylene 환원력(還元力)은 출수기에 0.012nmol/hour로 감소(減少)하였다가 수확기에 0.44nmol/g/hour로 증가(增加)하였으며 보릿짚시용(施用)에 의한 뚜렷한 차이(差異)는 없었다. 3. 효소활성(酵素活性)의 변화(變化)는 ${\beta}$-glucosidase가 출수기에 $426.8{\mu}mol/g/hour$로 크게 증가(增加)하였다가 그 후(後)에는 $71.64{\mu}mol/g/hour$로 감소(減少)하였으며 Phosphatase는 무재배구(無栽培區)에서 경시적(經時的)으로 감소(減少)하였으나 재배구(栽培區)에서는 경시적(經時的)으로 증가(增加)하였다. Protease는 출수기에 $73.1{\mu}mol/g/hour$로 증가(增加)하였으며 보릿짚시용(施用)에 의해 분얼기 무재배구(無栽培區)에서 $94.71{\mu}mol/g/hour$, 출수기 재배구(栽培區)에서 $73.1{\mu}mol/g/hour$로 증가(增加)하였다. 4. 토양중(土壞中) 당(糖)의 변화(變化)는 경시적(經時的)으로 감소(減少)하였으며 보릿짚시용(施用)에 의해 분얼기 재배구(栽培區)의 hexose가 $58.73{\mu}g/g$로 크게 증가(增加)되었다. 5. 유기물(有機物)의 형태(形態)는 ligno-protein이 재배구(栽培區)와 무재배구(無栽培區)에서 보릿짚시용(施用)에 의해 각각(各各) 3840.8mg/100g dry soil, 3898.6mg/100g dry soil로 가장 높았다.

• 한국동물위생학회지
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• 제20권3호
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• pp.297-306
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• 1997

This study was conducted to identify the effects of caffeine on the change of blood chemistry components in the serum of the rat(Sprague-Dawley, female). The experimental groups were divided into 7 groups according to the time lapsed after a single oral administration of 100mg/kg caffeine(that is control, 2, 4, 8, 24, 48 and 72 hrs lapsed group) to the rats. The concentrations of glucose, urea nitrogen, uric acid, creatinine, total protein, albumin, A/G ratio, triglyceride, total cholesterol, HDL-cholesterol, free fatty acid and 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 results obtained from this study were summarized as follows ; 1. The concentrations of serum glucose were significantly higher($\rho$<0.01) between 4 (143.0 mg/dl) and 8 hrs(138.0mg/dl) in comparison to the control(101.1mg/dl) after a single oral administration of caffeine(100mg/kg). Whereas there were no significant differences in the concentrations of urea nitrogen, uric acid, creatinine, total protein, albumin and albumin/globulin(A/G) ratio in comparison to the control. 2. The concentrations of total cholesterol and HDL-cholesterol in serum were significantly higher ($\rho$<0.01) between 4(77.4mg/dl, total cholesterol) and 8 hrs(64.7mg/dl, HDL-cholesterol) in comparis to the control(62.8, 46.7mg/dl) after a single oral administration of caffeine (100 mg/kg). On the other hand, the concentrations of triglyceride in serum were significantly lower($\rho$<0.01) after 8 hrs(38.8mg/dl) in comparison to the control(66.5mg/dl). 3. The activities of AST in serum was significantly higher($\rho$<0.05) from 2 hrs(149 U/L) to 8 hrs(178 U/L) in comparison to the control(112 U/L) after a single oral administration of caffeine (100mg/kg). The activities of ALT in serum were significantly higher($\rho$<0.01) at 4(45.5 U/L), 24(49.3 U/L), 48(46.8 U/L) and 72 hrs(42.3 U/L) in comparison to the control (39.7 U/L) after a single oral administration of caffeine (100mg/kg) On the other hand, there were no significant differences in the activities of ALP in comparison to the control.

• 대한수의학회지
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• 제36권4호
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• pp.795-807
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• 1996

This study was conducted to identify the effects of caffeine 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. Acute test were conducted to determine those effects. The acute test was conducted by dividing rats into 7 groups according to the time lapsed after a single oral administration of 100mg/kg caffeine(that is control, 2hrs, 4hrs, 8hrs, 24hrs, 48hrs and 72hrs lapsed group). 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, malondialdehyde(MDA) and the patterns of the SDS-PAGE(Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis) 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. The concentrations of serum glucose were significantly higher(p<0.01) between 4(143.0mg/dl) and 8hrs(138.0mg/dl) in comparison to that of the control(101.1mg/dl) after a single oral administration of caffeine(100mg/kg). While on the other, there were no significant differences in the concentrations of urea nitrogen, uric acid, creatinine, total protein, albumin and albumin/globulin(A/G) ratio in comparison to those of the control. 2. The concentrations of total cholesterol and HDL-cholesterol in serum were significantly higher(p<0.01) between 4(77.4mg/dl, total cholesterol) and 8hrs(64.7mg/dl, HDL-cholesterol) in comparison to those of the control(62.8, 46.7mg/dl) after a single oral administration of caffeine(100mg/kg). On the other hand, the concentrations of triglyceride in serum were significantly lower(p<0.01) after 8hrs(38.8mg/dl) in comparison to that of the control(66.5mg/dl). 3. The activities of AST in serum was significantly higher(p<0.05) from 2hrs(149U/L) to 8hrs(178U/L) in comparison to the control(112U/L) after a single oral administration of caffeine(100mg/kg). The activities of ALT in serum were significantly higher(p<0.01) at 4(45.5U/L), 24(49.3U/L), 48(46.8U/L) and 72 hrs(42.3U/L) in comparison to that of the control(39.7U/L) after a single oral administration of caffeine(100mg/kg). On the other hand, there were no significant differences in the activities of ALP in comparison to that of the control. 4. The concentrations of free fatty acid in serum were significantly higher(p<0.01) at 8hrs(65.0mg/dl) in comparison to that of the control(37.6mg/dl) after a single oral administration of caffeine(100mg/kg). However, there were no significant differences in the concentrations of carbonyl group and malondialdehyde within serum, and liver homogenate, mitochondrial and microsomal fractions in comparison to that of the control. 5. The patterns of SDS-PAGE in serum, mitochondrial and microsomal fraction of the liver showed no significant differences.

• Journal of Periodontal and Implant Science
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• 제29권1호
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• pp.173-192
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• 1999

The purpose of this study was to evaluate the effects of bioactive glass and natural coral on the human periodontal ligament fibroblast(HPLF) behaviors during the regeneration process of peridontium. To determine the cellular events occuring in the presence of the particles of bioactive glass and natural coral, HPLF were isolated from healthy premolar teeth extracted for orthodontic treatment. Cells were cultured in ${\alpha}$MEM at 37$^{\circ}C$, 5% $CO_2$, 95% humidity incubator. Bioactive glass and natural coral were powdered, and each particles(<40${\mu}$m) were placed on the cultured cells at the concentration of 0.3mg/ml, and 1,0mg/ml for experimental group. In control group no particles were added. And each group was evaluated by examining the cell morphology under phase-contrast micrograph at 4 day and transmission electron micrograph(TEM) and scanning electron micrograph(SEM) at 14 day, alkaline phosphatase activity at 5 and 9 day, protain synthesis at 4 day, DNA synthesis at 1, 2, 3 and 4 day, cell proliferation at 1, 3, 5,7 and 9 day and the formation of bone nodule at 30 day after culturing all groups in mineralizing supplemented mediun, No significant changes in cell morphology by adding these two matirials were found under phase contrast microscopy and TEM. HPLF phagocytocized each particles suggesting that HPLF is involved in the process of resorbing each particles and that bioactive glass were more biocompatible than natural coral. The ALPase activity of bioactive glass 0.3 mg/ml was similar with control groups and all the rests of control groups were significantly low(P<0.01) indicating a transient dedifferentiation of HPLF in the presence of bioactive glass and natural coral particles. There were no significant differences of protein synthesis between all groups. The DNA synthesis in experimental groups were significantly lower than control groups at 1, 2 and 3 day (P<0.01) but became similar to control groups at 4 day. Between control groups, the DNA synthesis in bioactive glass O.3mglml group was significantly higher than other groups(P<0.01). Cell proliferation in natural coral 1.0mg/ml and bioactive glass 1.0mglml groups were significantly lower than control group at 3 day(P<0.05) and there were no differences at 5, 7, 9 day. There were more bone nodule formation in experimental groups than in control groups. In conclusion, these results indicated that bioactive glass and natural coral have some effects of a transient dedifferentiation on HPLF and regeneration of periodontal tissues, however any significant cytotoxic effect on HPLF by these two particles were not found.

• 대한수의학회지
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• 제36권2호
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• pp.313-325
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• 1996

Experiments were undertaken to examine the ability of selenium to protect against alcohol and/or paraquat-induced hepatotoxicity and to examine the additive effect between alcohol and paraquat. Protective effect against hepatotoxic functions was measured in serum from alcohol(15% v/v), paraquat(200ppm), alcohol and paraquat, and combination of sodium selenite(4ppm) in drinking water-fed guinea pigs ad libitum for 4 weeks. A total of 68 healthy 7-weeks-old male animals were assigned at random to 8 treatment groups(9~13 animals/group). Body and liver weight losses, and high serum concentrations in aspartate aminotransferase(AST), alanine aminotransferase(ALT, in only paraquat group), $\gamma$-glutamyltranspeptidase($\gamma$-GTP), cholesterol(Cho), creatinine, blood urea nitrogen(BUN), total bilirubin(TB), direct bilirubin(DB), total protein(TP), albumin and globulin as well as low values in alkaline phosphatase(ALP) and glucose were produced in a groups of alcohol or paraquat-fed. These values were not potentiated in a group given the combination of alcohol plus paraquat. Morphological changes in the liver were also observed in the alcohol or paraquat-fed group. Lipid droplet and cell swelling in the hepatocytes were observed in alcohol-fed guinea pig, especially Mallory's hyaline arounded hepatic vein. In the paraquat-fed guinea pig, lipid droplet, pyknosis and karyolysis were observed. When alcohol or paraquat was combined with selenium-fed, hyperplasia of Kupffer cell in liver were observed. However, the mean ALT, $\gamma$-GTP, Cho, BUN, TB, TP, albumin and globulin values were lower in groups given the combination of alcohol and/or paraquat plus selenium, compared with groups given alcohol and/or paraquat. Also, the ratio of liver weight to body weight and ALP values(exception of paraquat plus selenium group) were increased by selenium. These results suggest that an adequate selenium confers marked protection against alcohol and paraquat-induced hepatotoxicity.