• Title/Summary/Keyword: COL8A2

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Periodontal healing using a collagen matrix with periodontal ligament progenitor cells in a dehiscence defect model in beagle dogs

  • Yoo, Seung-Yoon;Lee, Jung-Seok;Cha, Jae-Kook;Kim, Seul-Ki;Kim, Chang-Sung
    • Journal of Periodontal and Implant Science
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    • v.49 no.4
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    • pp.215-227
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    • 2019
  • Purpose: To histologically characterize periodontal healing at 8 weeks in surgically created dehiscence defects in beagle dogs that received a collagen matrix with periodontal ligament (PDL) progenitor cells. Methods: The bilateral maxillary premolars and first molars in 6 animals were used. Standardized experimental dehiscence defects were made on the buccal side of 3 premolars, and primary culturing of PDL progenitor cells was performed on the molars. Collagen matrix was used as a scaffold and a delivery system for PDL progenitor cells. The experimental sites were grafted with collagen matrix (COL), PDL progenitor cells with collagen matrix (COL/CELL), or left without any material (CTL). Histologic and histomorphometric analyses were performed after 8 weeks. Results: The defect height from the cementoenamel junction to the most apical point of cementum removal did not significantly differ across the CTL, COL, and COL/CELL groups, at $4.57{\pm}0.28$, $4.56{\pm}0.41$, and $4.64{\pm}0.27mm$ (mean ${\pm}$ standard deviation), respectively; the corresponding values for epithelial adhesion were $1.41{\pm}0.51$, $0.85{\pm}0.29$, and $0.30{\pm}0.41mm$ (P<0.05), the heights of new bone regeneration were $1.32{\pm}0.44$, $1.65{\pm}0.52$, and $1.93{\pm}0.61mm$ (P<0.05), and the cementum regeneration values were $1.15{\pm}0.42$, $1.81{\pm}0.46$, and $2.57{\pm}0.56mm$ (P<0.05). There was significantly more new bone formation in the COL/CELL group than in the CTL group, and new cementum length was also significantly higher in the COL/CELL group. However, there were no significant differences in the width of new cementum among the groups. Conclusions: PDL progenitor cells carried by a synthetic collagen matrix may enhance periodontal regeneration, including cementum and new bone formation.

Production of ColE1 Type Plasmid by Escherichia coli $DH5\alpha$ Cultured Under Nonselective Conditions

  • PASSARINHA L. A.;DIOGO M. M.;QUEIROZ J. A.;MONTEIRO G. A.;FONSECA L. P.;PRAZERES D. M. F.
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.20-24
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    • 2006
  • Plasmid DNA (pDNA) is a product of interest for many biopharmaceutical companies and research laboratories, because of increase in the number of gene therapy protocols that use nonviral vectors. This work was undertaken to study the effect of antibiotic and dissolved oxygen concentration (DOC) on the production of a ColE 1-type plasmid (pVAX1-LacZ) hosted in Escherichia coli $DH5\alpha$ and cultured in a batch fermentor with 0.751 of Terrific Broth. A decrease in the DOC from $60\%\;to\;5\%$ was shown to increase the specific pDNA concentration approximately 1.5-fold, due to the downregulation of growth. Additionally, this increase in the pDNA concentration led to a 2.2-fold increase in the purity of cell lysates obtained after cell lysis. However, the use of higher DOC led to 2.8-fold higher volumetric productivity as a consequence of a faster growth rate, reducing the fermentation time from 24 to 8 h. Interestingly, the specific pDNA concentration, and pDNA productivity and purity were always higher $(10-15\%)$ in the absence of antibiotic. Overall, the data indicate that nonselective conditions can be used without compromising yield, productivity, and purity of pDNA.

Development of an E. coli Expression Cassette for the Efficient Production of a Harmful Protein

  • Kim Ok Soo;Kwak Hwan Jong;Lee Jae-Hwa;Ha Jong Myung;Ha Bae-Jin;Lee Sang-Hyeon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.5
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    • pp.389-392
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    • 2004
  • In order to produce a harmful protein more efficiently, this expression cassette, dubbed pCol-MICT, is directed by the colicin promoter, and was constructed by the insertion of a $rrnBT_1T_2$ fragment of pEXP7, and a MxelnteinCBD fragment of pTXB3, into pSH375. To test whether harmful proteins, including proteolytic enzymes, could be effectively produced by this cassette, the carboxypeptidase (CPase) Taq gene was inserted into the pCol-MICT cassette to yield pCol-CPase Taq-MICT. E coli W3l 10 tells harboring pCol-CPase Taq-MICT produced a large quantity of this enzyme, as much as 47.2 mg of purified from per liter of culture, when cultured in the presence of mitomycin C ($0.4{\mu}g/mL$). This indicates that the colicin promoter-controlled E, coli expression cassette was able to produce almost 8 times of protein than the conventional tar promoter-based system, and that this cassette may be useful in the Synthesis of other harmful proteins.

A Study on the Dielectric and Pyroelectric Properties of the $Pb({Sn}_{1/2}Nb_{1/2})O_3-PbTiO_3-PhZrO_3$ Ceramics ($Pb({Sn}_{1/2}Nb_{1/2})O_3-PbTiO_3-PhZrO_3$ 세라믹의 유전 및 초전특성에 관한 연구)

  • Ham, Young-Wook;Lee, Neung-Hun;Kim, Yong-Hyuk;Kim, Jin-Soo;Kim, Young-Il
    • Proceedings of the KIEE Conference
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    • 1993.07b
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    • pp.1119-1121
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    • 1993
  • The PSN-PT-PZ ceramics doped with the Mn-oxide(0.5wt%) were fabricated by the mixed oxide method at 1250($^{\circ}C$) for 2(hr) and then the dielectric and pyroelectric properties were investigated with the compositions. In the 0.05PSN-0.4PT-0.55PZ specimen with 0.5(wt%) $MnO_2$, the pyroelectric coefficient was $6.6{\times}10^{-8}(C/Cm^2^{\circ}C)$, and the figure of merits for pyroelectric current and voltage were $27{\times}10^{-9},\;45{\times}10^{-12}$(C.Cm/J), respectibly.

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인공 항문계를 이용한 대두박 isoflavones의 생체 이용성 평가

  • 김경진;지규만
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2000.11a
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    • pp.91-93
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    • 2000
  • Soybean meal(SMB) has not only been an important source of protein, but also a major source of isolfavones(IF) for poultry diet. Since the IF is one of effective phytoestrogens, evaluation of bioavailability of the phytochemicals for poultry and development of an animal model for assessment of the availability should be essential for its practical application. Present study was to develop bird model for assessing their bioavailability. No reports for such measurments with poultry could have every been found. Eight, 6-wk-old, layer-type, male birds were divided into two groups of four birds; normal (N) and colostomized(COL). Each bird was tube-fed a SMB sample at a level of 1% body weight. Samples from blood, urine and feces(COL birds) or urine+feces (N) were collected at 0, 2, 8, 15, 24 and 48 hours after the feeding. Plasma and urine samples were treated with -glucuronidase and fecal samples with 4N HCl for measurement of genistein(GE) and daidzein (DA) with HPLC set with an electrochemical detector. Maximum peaks of plasma IF appeared at 2 h after the meal for both birds groups. However the IF peak for N group was GE, while that for COL group was DA. Plasma DA levels for COL birds were always higher than those of GE, which was in opposite trend for N birds. This trend observed in the N birds is in accordance with those reported in rats fed SBM. In conclusion the results seem to suggest that colostomy as an attempt to separate urine from feces in birds affected the IF metabolism in the intestine and may not be a proper bird model to measure the IF bioavailability.

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The complete mitochondrial genome of Arabidopsis thaliana (Brassicaceae) isolated in Korea

  • PARK, Jongsun;XI, Hong;KIM, Yongsung
    • Korean Journal of Plant Taxonomy
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    • v.51 no.2
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    • pp.176-180
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    • 2021
  • Arabidopsis thaliana (L.) Heynh. is a small plant species that serves as a model organism of plant biology and genetics. Here, we present the first complete mitochondrial genome of Korean A. thaliana natural isolate (named as 180404IB4), which is 368,875 bp long and contains 58 genes (33 protein-coding genes, 22 tRNAs, and three rRNAs), with a GC ratio of 44.8%. Sixty-four single-nucleotide polymorphisms and 11 insertion and deletion regions (1,089 bp in length) are identified against the Col-0 ecotype, showing one large insertion of 1,069 bp without structural variation. Phylogenetic trees constructed from 30 conserved genes indicate that the 180404IB4 mitochondrial genome is clustered with Col-0 and three East Asian ecotypes.

Effects of 2-deoxy-D-glucose and quercetin on cytokine secretion and gene expression of type I collagen during osteoblastic differentiation in irradiated MC3T3-El cells

  • Song Haeng-Un;Ahn Hyoun-Suk;Lee Sang-Rae;Koh Kwang-Joon
    • Imaging Science in Dentistry
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    • v.35 no.4
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    • pp.191-198
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    • 2005
  • Purpose: To characterize the effects of 2-deoxy-D-glucose (2DG) and quercetin (QCT) on cytokine secretion of IL-6, $TGF-\beta$ and gene expression of Col I in irradiated MC3T3-E1 cells Materials and Methods: The MC3T3-El cells were cultured in an a-MEM supplemented with 5mM 2DG or 10mM QCT and then the cells were incubated 12h before irradiation with 2, 4, 6, and 8Gy X-ray using a linear accelerator delivered at a dose rate of 1.5Gy/min. Level of IL-6 and $TGF-\beta$ was determined by ELISA. Also expression of Col I was examined by RT-PCR. Results: In accordance with the radiation dose, the amount of $TGF-\beta$ was not different in RA + QCT, but it showed a peak value in control and RA + 2DG at 4Gy on the 3rd day. However, all groups showed a decreasing tendency dose-dependently in RA+QCT on the 7th day (p<0.01). In accordance with the radiation dose, the amount of IL-6 increased dose-dependently in all groups on the 3rd day. On the 7th and 21st day, all groups showed peak values at 4Gy. RA+QCT showed a slightly increased amount of IL-6 at 2Gy, but it showed a slightly decreased amount at 4, 6, and 8Gy. In accordance with the period of culture after irradiation, the expression of Col I increased dose-dependently in RA+QCT. Conclusion: The result showed that QCT acted as radiosensitizer in the secretion of $TGF-\beta$ and gene expression of Col I during differentiation in irradiated MC3T3-E1 cells at the cellular level.

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Establishment and Application of Molecular Genetic Techniques for Preimplantation Genetic Diagnosis of Osteogenesis Imperfecta (골형성부전증의 착상전 유전진단을 위한 분자유전학적 방법의 조건 확립과 적용)

  • Kim, Min-Jee;Lee, Hyoung-Song;Choi, Hye-Won;Lim, Chun-Kyu;Cho, Jae-Won;Kim, Jin-Young;Song, In-Ok;Kang, Inn-Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.35 no.2
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    • pp.99-110
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    • 2008
  • Objectives: Preimplantation genetic diagnosis (PGD) has become an assisted reproductive technique for couples carrying genetic conditions that may affect their offspring. Osteogenesis imperfecta (OI) is an autosomal dominant disorder of connective tissue characterized by bone fragility and low bone mass. At least 95% of cases are caused by dominant mutations in the COL1A1 or COL1A2. In this study, we report on our experience clinical outcomes with 5 PGD cycles for OI in two couples. Methods: Before clinical PGD, we assessed the amplification rate and allele drop-out (ADO) rate of alkaline lysis and nested PCR protocol using heterozygous patient's single lymphocytes in the pre-clinical diagnostic tests for OI. We performed 5 cycles of PGD for OI by nested PCR for the causative mutation loci, COL1A1 c.2452G>A and c.3226G>A, in case 1 and case 2, respectively. The PCR products were analyzed by agarose gel electrophoresis, restriction fragment length polymorphism (RFLP) analysis with HaeIII restriction enzyme in the case 1 and direct DNA sequencing. Results: We confirmed the causative mutation loci, COL1A1 c.2452G>A in case 1 and c.3226G>A in case 2. In the pre-clinical tests, the amplification rate was 94.2% and ADO rate was 22.5% in case 1, while 98.1% and 1.9% in case 2, respectively. In case 1, a total of 34 embryos were analyzed and 31 embryos (91.2%) were successfully diagnosed in 3 PGD cycles. Eight out of 19 embryos diagnosed as unaffected embryos were transferred in all 3 cycles, and in the third cycle, pregnancy was achieved and a healthy baby was delivered without any complications in July, 2005. In case 2, all 19 embryos (100.0%) were successfully diagnosed and 4 out of 11 unaffected embryos were transferred in 2 cycles. Pregnancy was achieved in the second cycle and the healthy baby was delivered in March, 2008. The causative locus was confirmed as a normal by amniocentesis and postnatal diagnosis. Conclusions: To our knowledge, these two cases are the first successful PGD for OI in Korea. Our experience provides a further demonstration that PGD is a reliable and effective clinical techniques and a useful option for many couples with a high risk of transmitting a genetic disease.

Genetic analysis using whole-exome sequencing in pediatric chronic kidney disease: a single center's experience

  • Lee, Hyeonju;Min, Jeesu;Ahn, Yo Han;Kang, Hee Gyung
    • Childhood Kidney Diseases
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    • v.26 no.1
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    • pp.40-45
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    • 2022
  • Purpose: Chronic kidney disease (CKD) has various underlying causes in children. Identification of the underlying causes of CKD is important. Genetic causes comprise a significant proportion of pediatric CKD cases. Methods: In this study, we performed whole-exome sequencing (WES) to identify genetic causes of pediatric CKD. From January to June 2021, WES was performed using samples from pediatric patients with CKD of unclear etiology. Results: Genetic causes were investigated using WES in 37 patients (17 males) with pediatric CKD stages 1 (n=5), 2 (n=7), 3 (n=2), 4 (n=2), and 5 (n=21). The underlying diseases were focal segmental glomerulosclerosis (n=9), congenital anomalies of the kidney and urinary tract including reflux nephropathy (n=8), other glomerulopathies (n=7), unknown etiology (n=6), and others (n=7). WES identified genetic causes of CKD in 12 of the 37 patients (32.4%). Genetic defects were discovered in the COL4A4 (n=2), WT1 (n=2), ACTN4, CEP290, COL4A3, CUBN, GATA3, LAMA5, NUP107, and PAX2 genes. WT1 defects were found in patients whose pathologic diagnosis was membranoproliferative glomerulonephritis, and identification of CUBN defects led to discontinuation of immunosuppressive agents. Genetic diagnosis confirmed the clinical diagnosis of hypoparathyroidism, sensorineural deafness, and renal disease; Alport syndrome; and Joubert syndrome in three of the patients with CKD of unknown etiology (COL4A4 [n=2], CUBN [n=1]). Extrarenal symptoms were considered phenotypic presentations of WT1, PAX2, and CEP290 defects. Conclusions: WES provided a genetic diagnosis that confirmed the clinical diagnosis in a significant proportion (32.4%) of patients with pediatric CKD.

An epidemiologic study on the cleft lip and/or palate patients who visited Dept. of Orthodontics, Pusan National University Hospital (부산대학교병원 교정과에 내원한 구순구개열 환자들에 대한 역학조사)

  • Son Woo-Sung;Baek Jae-Ho;Lee Won-Chul
    • Korean Journal of Cleft Lip And Palate
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    • v.5 no.1
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    • pp.43-58
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    • 2002
  • A General survey on the cleft lip and/or palate patients in Pusan National University Hospital was done. The conclusions were as follow. 1. In gender ratio, male was higher than female. 2. In the cleft type, unilateral cleft lip and palate, cleft lip and alveolus, and bilateral cleft lip and palate in higher ratio order. Left unilateral cleft lip and palate was higher than right in ratio. 3. In first hospital visiting age for dental treatment, the group from 6 years old to 12 years old was most and in that the age group for attending elementary school (about 7-8 years old) showed especially higher ratio. The large majority of patient were born in 1980's and lived in Pusan metropolitan city, KyungSangNamDo. Especially, most of them lived in the neighboring Held of Pusan National University Hospital. 4. Class III skeletal pattern and Angle's Class III molar relationship was most prevalent. Angle's Class II molar relationship showed relative higher ratio because tooth loss and malposition. 5. Primary lip closure in 3 months, secondary lip closure in 6-7 years old, palatal closure in 12-16 months and bone graft in 8-10 years old were operated mostly. Rhinoplasty and scar revision were operated in too early age so the need of infantile orthopedics must be considered. 6. Maxillary expansion and protraction were operated mostly in 8-11 years old but operating in primary dentition must be considered. 7. Tooth alignment were started mostly in mixed dentition and consideration about prosthodontic treatment and retention will be need. 8. In tooth anomaly, tooth malformation and missing were most prevalent.

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