• International Journal of Oral Biology
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• v.33 no.1
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• pp.33-43
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• 2008

In the process of bone remodeling, mineral phase of bone is dissolved by osteoclasts, resulting in elevation of calcium concentration in micro-environment. This study was performed to explore the effect of high extracellular calcium ($Ca{^{2+}}_e$) on mineralized nodule formation and on the expression of progressive ankylosis (Ank), plasma cell membrane glycoprotein-1 (PC-1) and osteopontin by primary cultured mouse calvarial cells. Osteoblastic differentiation and mineralized nodule formation was induced by culture of mouse calvarial cells in osteoblast differentiation medium containing ascorbic acid and ${\beta}$-glycerophosphate. Although Ank, PC-1 and osteopontin are well known inhibitors of mineralization, expression of these genes were induced at the later stage of osteoblast differentiation during when expression of osteocalcin, a late marker gene of osteoblast differentiation, was induced and mineralization was actively progressing. High $Ca{^{2+}}_e$(10 mM) treatment highly enhanced mRNA expression of Ank, PC-1 and osteopontin in the late stage of osteoblast differentiation but not in the early stage. Inhibition of p44/42 MAPK activation but not that of protein kinase C suppressed high $Ca{^{2+}}_{e^-}$induced expression of Ank, PC-1 and osteopontin. When high $Ca{^{2+}}_e$(5 mM or 10 mM) was present in culture medium during when mineral deposition was actively progressing, matrix calcifiation was significantly increased by high $Ca{^{2+}}_e$. This stimulatory effect was abolished by pyrophosphate (5 mM) or levamisole (0.1-0.5 mM), an alkaline phosphatase inhibitor. In addition, probenecid (2mM), an inhibitor of Ank, suppressed matrix calcification in both control and high $Ca{^{2+}}_{e^-}$treated group, suggesting the possible role of Ank in matrix calcification by osteoblasts. Taken together, these results showed that high $Ca{^{2+}}_e$ stimulates expression of Ank, PC-1 and osteopontin as well as matrix calcification in late differentiation stage of osteoblasts and that p44/42 MAPK activation is involved in high $Ca{^{2+}}_{e^-}$induced expression of Ank, PC-1 and osteopontin.

• Journal of the Korean Academy of Child and Adolescent Psychiatry
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• v.1 no.1
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• pp.161-171
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• 1990

The study was to examine the effectiveness of the educational program for the parents of autistic children in promoting the development and amelioration of psychopathology. Behavioral pathology and developmental function of autistic children of two parent groups, parents who participated in educational program and parents who did not participate in that program, were compared before and after partent education program. For these purpose, the data were collected from 30 subjects(15 for participant group and 15 for non-participant group) who were diagnosed as Autism according to DSM-III at Child-Psychiatry of Seoul National University Hospital during May 16, 1987 through April 30, 1988. Pre and post tests by Schopler's Psycho-Educational Profile scale were performed for all of the subjects and the data were analyzed by Wilcoxon Rank Sum test, Wilcoxon Sign Rank test and $X^2-test.$ The results obtained were as follows ; 1) The autistic children of participant group were significantly higher than those of non-participant group after education on Imitation, Gross motor and Eye-hand Integration scale of developmental function area (p<0.05) 2) After education, the autistic children of participant group were significantly improved in Affect and Language scale of behavioral pathology area (p<0.05). 3) After education, the autistic children of participant group were significantly improved in developmental function, such as perception, Fine motor, Gross motor, Eye-hand Integration, Cognitive Performance, Cognitive Language (p<0.05). 4) The autistic children of non-participant group were significantly improved in Relating behavior and developmental functions such as Perception, Fine motor and Eye-hand Integration at post test in comparison to pre test (p<0.05). From these results, it might be concluded that the autistic children of participant group were improved much more, and obtained higher score in developmental function area than those of non-participant group. It is suggested that this kind of education program helped and supported the parents to do some more appropriate approach for the development of their children.

• International Journal of Oral Biology
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• v.32 no.3
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• pp.85-91
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• 2007

Dlx3 is a homeodomain protein and is known to playa role in development and differentiation of many tissues. Deletion of four base pairs in DLX3 (NT3198) is causally related to tricho-dento-osseous (TDO) syndrome (OMIM # 190320), a genetic disorder manifested by taurodontism, hair abnormalities, and increased bone density in the cranium. Although the observed defects of TDO syndrome involves bone, little is known about the role of Dlx3 in bone remodeling process. In this study, we examined the effect of wild type DLX3 (wtDlx3) expression on osteoclast differentiation and compared it with that of 4-BP DEL DLX3 (TDO mtDlx3). To examine whether Dlx3 is expressed during RANKL-induced osteoclast differentiation, RAW264.7 cells were cultured in the presence of receptor activator of nuclear factor-B ligand (RANKL). Dlx3 protein level increased slightly after RANKL treatment for 1 day and peaked when the fusion of prefusion osteoclasts actively progressed. When wtDlx3 and TDO mtDlx3 were overexpressed in RAW264.7 cells, they enhanced RANKL-induced osteoclastogenesis and the expression of osteoclast differentiation marker genes such as calcitonin receptor, vitronectin receptor and cathepsin K. Since osteoclast differentiation is critically regulated by the balance between RANKL and osteoprotegerin (OPG), we examined the effect of Dlx3 overexpression on expression of RANKL and OPG in C2C12 cells in the presence of bone morphogenetic protein 2. Overexpression of wtDlx3 enhanced RANKL mRNA expression while slightly suppressed OPG expression. However, TDO mtDlx3 did not exert significant effects. This result suggests that inability of TDO mtDlx3 to regulate expression of RANKL and OPG may contribute to increased bone density in TDO syndrome patients. Taken together, it is suggested that Dlx3 playa role as a positive regulator of osteoclast differentiation via up-regulation of osteoclast differentiation-associated genes in osteoclasts, as well as via increasing the ratio of RANKL to OPG in osteoblastic cells.

• Journal of Embryo Transfer
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• v.33 no.3
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• pp.139-147
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• 2018

Somatic cell nuclear transfer (SCNT) is a useful biotechnological tool for animal cloning. Until now, SCNT has been inefficient, especially in dog. It is believed that an embryo developmental block in SCNT embryos is cause of low production efficiency. However, no studies have been performed on canines for embryo developmental block. In this study, we attempted to evaluate the beneficial role of EDTA in canine parthenogenic (PA) embryos development to overcome embryo developmental block. The PA embryos were divided into 0.01 mM EDTA treated and non-treated groups. Embryo developmental efficiency was measured by activating chemically parthenote. After EDTA induction, PA embryos were evaluated for embryonic development, Reactive Oxygen Species (ROS) activity, mitochondrial integrity, ATP production and genomic activation. The EDTA treated PA embryos showed significantly higher survival rate and improved cavity formation compared to non-treated. Furthermore, cytoplasmic ROS level was mitigated and mitochondrial membrane potential was found significantly higher in EDTA treated group followed by higher ATP production. Moreover, major embryonic genomic activation specific markers/factors were also elevated in EDTA treated group. Conclusively, we elucidated that EDTA showed substantially positive effect to overcome embryo developmental block in canine.

• Korean Journal of Veterinary Research
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• v.40 no.3
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• pp.421-430
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• 2000

The regional distribution and relative frequency of the pancreatic polypeptide (PP)-immunoreactive cells in the pancreas of the Korean native goat (Capra hircus) were studied by immunohistochemical methods (PAP methods) using specific antisera against to PP during developmental stage, fetus, neonate, 1-month-old, 6-month-old and adult. The different regional distribution and relative frequency of PP-immunoreactive cells in the pancreas of Korean native goat were observed during development. In the exocrine portions, they were detected in the all ages and distributed in the interacinar regions. PP-immunoreactive cells were increased from neonate to 1-month-old but thereafter decreased with developmental stage. In the pancreatic duct, PP-immunoreactive cells were demonstrated from 1-month-old but they were decreased with developmental stages in these regions. These cells were observed in the subepithelial connective tissue of the pancreatic duct with a few frequencies in 1-and 6-month-old. However, they were located in the interepithelial cells of the pancreatic duct in the adult with rare frequency. In the endocrine portion (pancreatic islets), PP-immunoreactive cells were observed from neonate, and the regional distribution of PP-immunoreactive cells from 1-month-old was divided into two patterns : dispersed 1) in the marginal regions of the pancreatic islets with moderate or a few frequencies, and 2) in the whole pancreatic islets (in a case of 1-month-old) or in the central cores (in a case of 6-month-old and adult) with numerous frequency. In conclusion, the different regional distribution and relative frequency of PP-immunoreactive cells in the pancreas of the Korean native goat were observed during development. It is suggested that the changing of feeding habits and physiological conditions during different developmental stages may cause these differences.

• Development and Reproduction
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• v.14 no.2
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• pp.59-63
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• 2010

This study was performed to examine the influence of water temperature on egg developmental speed for determining the required time and optimum water temperature for hatching of olive flounder Paralichthys olivaceus eggs. The fertilized eggs were collected from the naturally spawned adults in November 2007. The eggs were randomly divided into 6 groups of temperature (5, 10, 15, 20, 25 and $30^{\circ}C$) and transferred in $1{\ell}$ beaker, respectively. The fertilized eggs of the olive flounder did not hatched at $5^{\circ}C$ and $30^{\circ}C$ and hatching rates at 10, 15, 20 and $25^{\circ}C$ were 3, 12, 25 and 50%, respectively. The relationships between the water temperature (T, $^{\circ}C$) and required time (1/t, hour) from egg to each developmental stage were given as follows ; Blastula: 1/t=0.0208T-0.0951 ($r^2$=0.8593) Kupffer's vesicle: 1/t=0.0052T-0.0176 ($r^2$=0.9819) Myotome: 1/t=0.0034T-0.0172 ($r^2$=0.8508) Hatching: 1/t=0.0016T-0.0068 ($r^2$=0.9915) Biological minimum temperature in egg development was calculated to be $4.3^{\circ}C$.

• The Korean Journal of Pesticide Science
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• v.4 no.4
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• pp.72-76
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• 2000

The study was conducted to investigate the developmental periods and effect of several insecticides on Hemiptarsenus sp., ectoparasitoid of Liriomyza trifolii. The mean length and width of egg were 0.5mm and 0.1mm. The mean length of larva, pupae, abult female, and abult male were 1.9mm, 2.0mm, 2.2mm, and 1.8mm, respectively. Developmental periods of Hemiptarsenus sp. from egg to larva at 15, 20, 25, $30^{\circ}C$ were 16.9, 8.8, 5.9, and 4.5 days, and those of pupa were 20.7, 9.7, 5.6, and 3.4 days, respectively. Based on these results, developmental threshold temperatures and effective temperatures were $9.5^{\circ}C$, 91.5 degree-days in egg-larval stage, $10.9^{\circ}C$, 142.3 degree-days in pupal stage. When several insecticides were evaluated to Hemiptarsenus sp. at the recommended concentrations, B.t WP, diflubenzuron WP, and cyromazin were negligiblly effective all life stages. Fipronil SC, cartap SP, spinosad GW were less toxic to larva and pupa, but highly toxic to adults. Abamectin EC was less toxic to all life stages, but inhibited oviposition of 50% more to Hemiptarsenus sp. female.

• Journal of Embryo Transfer
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• v.16 no.2
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• pp.139-144
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• 2001

This study was carried out to investigate the effects of the bull, sperm type and sperm pretreatment on the pronuclear formation and in vitro development after injection of spermatozoa into in vitro matured bovine oocytes. 1. Spermatozoa derived room four bulls(A, B, C and D) were used for ICSI. The male pronuclear formation and developmental rates were 73.9∼87.0% and 33.3∼60.9%, respectively. 2. The effects of sperm type were examined. Male pronuclear formation rates by using fresh-and frozen-sperm, tail-cutting and tail-scoring sperm were 82.0%. 78.0%. 42.2% and 51.1% (p<0.05) while development rates were 56.0%. 42.0%, 17.8% and 22.2%, respectively. Fresh sperm achived a high mail pronuclear- and development rates than those of other groups. 3. Cheroical pretreatments were tested and compared. When sperm were pretreated with heparin, BFF(bovine follicula fluid), His, Ca Ionophore(I) and I + caffeine, mate pronuclear formation and developmental rates were 66.7∼82.2% and 33.3∼60.6%. respectively. and these values of treatment of I + caffeine were higher than that of other methods. 4. The male pronuclear formation and developmental rates of oocytes obtained by ICSI treated with or without zona pellucida were 80.0%. 72.0% and 46.0%, 36.0%, respectively.

• International Journal of Oral Biology
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• v.34 no.1
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• pp.21-28
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• 2009

Mutations in DLX3 are associated with both autosomal dominant hypoplastic hypomaturation amelogenesis imperfecta (ADHHAI) and tricho-dento-osseous (TDO) syndrome. ADHHAI is caused by a c.561_562delCT (2bp-del DLX3) mutation whereas TDO syndrome is associated with a c.571_574delGGGG (4bp-del DLX3) mutation. However, although the causal relationships between DLX3 and an enamel phenotype have been established, the pathophysiological role of DLX3 mutations in enamel development has not yet been clarified. In our current study, we prepared expression vectors for wild type and deletion mutant DLX3 products (4bp-del DLX3, 2bp-del DLX3) and examined the effects of their overexpression on the expression of the enamel matrix proteins and proteases. Wild type DLX3 enhanced the expression of matrix metalloprotease 20 (MMP20) mRNA and protein in murine ameloblast-like cells. However, neither a 4bp-del nor 2bp-del DLX3 increased MMP20 expression. Wild type DLX3, but not the above DLX3 mutants, also increased the activity of reporters containing 1.5 kb or 0.5 kb of the MMP20 promoter. An examination of protein stability showed that the half-life of wild type DLX3 protein was less than 12 h whilst that of both deletion mutants was longer than 24 h. Endogenous Dlx3 was also found to be continuously expressed during ameloblast differentiation. Since inactivating mutations in the gene encoding MMP20 are associated with amelogenesis imperfecta, the inability of 4bp-del or 2bp-del DLX3 to induce MMP20 expression suggests a possible involvement of such mutations in the enamel phenotype associated with TDO syndrome or ADHHAI.

• International Journal of Oral Biology
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• v.32 no.4
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• pp.119-125
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• 2007

Dlx3 is a homeodomain protein and is known to play a role in development and differentiation of many tissues. Deletion of four base pairs in DLX3 (NT3198) is causally related to tricho-dento-osseous (TDO) syndrome (OMIM #190320), a genetic disorder manifested by taurodontism, hair abnormalities, and increased bone density in the cranium. The molecular mechanisms that explain the phenotypic characteristics of TDO syndrome have not been clearly determined. In this study, we examined phenotypic characteristics of wild type DLX3(wtDlx3) and 4-BP DEL DLX3 (TDO mtDlx3) in C2C12 cells. To investigate how wtDlx3 and TDO mtDlx3 differentially regulate osteoblastic differentiation, reporter assays were performed by using luciferase reporters containing the promoters of alkaline phosphatase, bone sialoprotein or osteocalcin. Both wtDlx3 and TDO mtDlx3 enhanced significantly all the reporter activities but the effect of mtDlx3 was much weaker than that of wtDlx3. In spite of these differences in reporter activity, electrophoretic mobility shift assay showed that both wtDlx3 and TDO mtDlx3 formed similar amounts of DNA binding complexes with Dlx3 binding consensus sequence or with ALP promoter oligonucleotide bearing the Dlx3 binding core sequence. TDO mtDlx3 exhibits a longer half-life than wtDlx3 and it corresponds to PESTfind analysis result showing that potential PEST sequence was missed in carboxy terminal of TDO mtDlx3. In addition, co-immunoprecipitation demonstrated that TDO mtDlx3 binds to Msx2 more strongly than wtDlx3. Taken together, though TDO mtDlx3 acted as a weaker transcriptional activator than wtDlx3 in osteoblastic cells, there is possibility that during in vivo osteoblast differentiation TDO mtDlx3 may antagonize transcriptional repressor activity of Msx2 more effectively and for longer period than wtDlx3, resulting in enhancement of osteoblast differentiation.