• Title/Summary/Keyword: DNA quantity

Search Result 111, Processing Time 0.026 seconds

Investigation of Nanofiber and Thermosensitive Scaffold for Intervertebral Disc through Organ Culture (기관배양을 통한 추간판 재생용 나노파이버 및 온도 감응성 지지체에 대한 검증)

  • Lee, Yong-Jae;Shin, Ji-Won;Shin, Ho-Jun;Kim, Chan-Hwan;Park, Ki-Dong;Bae, Jin-Woo;Seo, Hyoung-Yeon;Kim, Young-Jick;Shin, Jung-Woog
    • Journal of Biomedical Engineering Research
    • /
    • v.28 no.4
    • /
    • pp.512-519
    • /
    • 2007
  • The purpose of this study is to investigate the potential of a novel tissue engineering approach to regenerate intervertebral disc. In this study, thermosensitive scaffold (chitosan-Pluronic hydrogel) and nanofiber were used to replace the nucleus pulposus (NP) and annulus fibrosus of a degenerated intervertebral disc, leading to an eventual regeneration of the disc using the minimally invasive surgical procedure and organ culture. In preliminary study, disc cells were seeded into the scaffolds and cellular responses were assessed by MTT assay and scanning electron microscopy (SEM). Based on these results, we could know that tissue engineered scaffolds might provide favorable environments for the regeneration of tissues. Organ culture was performed in fresh porcine spinal motion segments with endplates on both sides. These spinal motion segments were classified into three groups: control (Intact), injured NP (Defect), and inserting tissue engineered scaffolds (Insert). The specimens were cultivated for 7 days, subsequently structural stability, cell proliferation and morphological changes were evaluated by the relaxation time, quantity of DNA, GAG and histological examination. In these results, inserting group showed higher relaxation time, reduced decrement of DNA contents, and accumulated GAG amount. Consequently, the tissue engineered scaffolds used in this study seen to be a promising base scaffolds for regenerative intervertebral disc due to its capacity to absorb external dynamic loading and the possible ideal environment provided for disc cell growing.

Detection of Hepatitis B Virus Using Micro-PCR and Real-Time PCR Methods (Micro-PCR과 Real-Time PCR을 이용한 B형 간염 바이러스 검출)

  • Kang, Won;Park, Sang-Bum;Nam, Youn-Hyoung;An, Young-Chang;Lee, Sang-Hyun;Jang, Won-Cheoul;Park, Su-Min;Kim, Jong-Wan;Chong, Song-Chun
    • Journal of the Korean Chemical Society
    • /
    • v.51 no.1
    • /
    • pp.36-42
    • /
    • 2007
  • Hepatitis B is a serious public health problem leading to chronic infection and liver cancer. Quantitation of circulating hepatitis B virus (HBV) is important for monitoring disease progression and for assessing the response to antiviral therapy. In this study, by using Real-Time PCR and novel Micro-PCR assay method, we measured HBV concentration in the clinical sample. A total of 120 serum samples from patients with HBV infection collected was in Dankook university hospital to compare the detection limit, sensitivity, specificity and reproducibility of the two assay methods. These findings of this study suggest that Micro-PCR and Real-Time PCR assay methods are comparable to each other in there detection limit, sensitivity, and reproducibility for HBV DNA quantitation. However, Micro-PCR assay is more efficient than Real-Time PCR method, because Real-Time PCR is not so time - consuming, technically easy and need to reagent of a small quantity. It will be useful for rapid and reliable clinical diagnosis of HBV in many countries.

Chromosomal Localization of Korean Cattle (Hanwoo) BAC Clones via BAC end Sequence Analysis

  • Chae, Sung-Hwa;Kim, Jae-Woo;Choi, Jae Min;Larkin, Denis M.;Everts-van der Wind, Annelie;Park, Hong-Seog;Yeo, Jung-Sou;Choi, Inho
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.20 no.3
    • /
    • pp.316-327
    • /
    • 2007
  • In this study, a Korean native cattle strain (Hanwoo) evidencing high performance in terms of both meat quality and quantity was employed in the generation of 150,000 BAC clones with an average insert size of 140 kb, and corresponding to about a 6X coverage of bovine chromosomal DNA. The BAC clones were pooled in a mini-scale via three rounds of a pooling protocol, and the efficiency of this pooling protocol was evaluated by testing the accuracy of accessibility to the positive clones, via a PCR-based screening method. Two sets of primers designed from each of two known genes were tested, and each yielded 2 or 3 positive clones for each gene, thereby indicating that the BAC library pooling system was appropriate with regard to the accession of the target BAC clones. Analyses of $3.3{\times}10^6$ base pairs obtained from the 7,090 BAC end sequence (BES) showed that 34.88% of the DNA sequence harbored the repetition sequence. Analysis of the 7,090 BES to the $1^{st}$ and $2^{nd}$ generation radiation hybrid map of the cattle genome, using the COMPASS program designed for the construction of a cattle-human comparative mapping, resulted in the localization of a total of 1,374 clones proximal to 339 $1^{st}$ generation markers, and 1,721 clones proximal to 664 $2^{nd}$ generation markers. Collectively, the BAC library and pooling system of the BAC clones from the Korean cattle, coupled with the chromosome-localized BAC clones, will provide us with novel tools for the excavation of desired clones for genome mapping and sequencing, and will also furnish us with additional information regarding breed differences in cattle.

Characteristics and breeding of a new cultivar Pleurotus eryngii, Seolsong (큰느타리버섯 신품종 '설송'의 육성 및 그 특성)

  • Shin, Pyung-Gyun;Yoo, Young-Bok;Kong, Won-Sik;Jang, Kab-Yeul;Oh, Youn-Lee;Cheong, Jong-Chun
    • Journal of Mushroom
    • /
    • v.11 no.2
    • /
    • pp.77-81
    • /
    • 2013
  • To develop a new cultivar of king oyster mushroom(Pleurotus eryngii), G09-21 as parental strain was selected by the method of Di-mon crossing between monokaryotic strain ASI 2824-21 derived from ASI 2824(Keunneutari No. 2) and dikaryotic strain ASI 2887(Aeryni 3). The Pe21-53(G09-21-10 x ASI 2844-9) was shown the best cultural characteristics, selected to be a new cultivar and designated as 'Seolsong'. The 'Seolsong' was distinctly formed incompatibility line in the confrontation growth of parental strains Keunneutari No. 2, Aeryni 3 and ASI 2844. Analysis of the genetic characteristics of the new cultivar 'Seolsong' showed a different DNA profile as that of the control strains, Keunneutari No. 2, Aeryni 3 and ASI 2844, when RAPD(Random Amplified Polymorphic DNA) primer URP4 was used. The optimum temperature and pH arrange for mycelial growth were $25{\sim}30^{\circ}C$ and pH 5~8, respectively. This new cultivar 'Seolsong' of fruiting body production per bottle was about $131.4{{\pm}}43.1$ g which is about 102% quantity compared to that of other cultivar Keunneutari No. 2. And also the stipe is thick and long, but the number of available stipe is few. Particularly, it was tolerant of high moisture above 90% during the growth period after primodia formation. We therefore expect that this new strain will save of labor and cost of cultivation by without culling work.

Phylogenetic characterization of bacterial populations in different layers of oak forest soil (상수리나무림의 토양 층위별 세균군집의 계통학적 특성)

  • Han, Song-Ih
    • Korean Journal of Microbiology
    • /
    • v.51 no.2
    • /
    • pp.133-140
    • /
    • 2015
  • We have examined the correlation between the physicochemical and microbiological environment variables for the different layers of oak forest soil in Mt. Gyeryong, Korea. The result shows that there is a high correlation in the environment variables between the soil parameters of the fermented (F) layer and humus (H) layer. In particular, the pH level in the F layer shows a high correlation with C and N, while the various organic acids of the H layer turns out to be closely correlated with soil bacteria density. As we evaluated phylogenetic characteristics of bacterial populations by DGGE analysis with DNA extracted. Total of 175 bands including 43 bands from litter (L) layer, 42 bands from F layer, 43 bands from H layer and 47 bands from rhizosphere (A) layer were selected as the major DGGE band of oak forest soil. Based on the 16S rRNA gene sequences, 175 DGGE bands were classified into 32 orders in 7 phylum. The heat map was analyzed in order to compare the quantity of the base sequences of each order and based on the clustering of the different layers of oak forest soil, the result confirms that the F layer and H layer belong to a different cluster from that of L layer and A layer. Furthermore, it also showed that approximately 50% of the total microbial population in different layers is ${\alpha}$-proteobacteria, which indicates that they belong to the dominant system group. In particular, Rhizobiales, Burkholderiales and Actinobacteriales were observed in all the seasons and layers of oak forest soil, which confirms that they are the indigenous soil bacterial community in oak forest soil.

Molecular Survey of Latent Pseudorabies Virus Infection in Nervous Tissues of Slaughtered Pigs by Nested and Real-time PCR

  • Yoon Hyun A;Eo Seong Kug;Aleyas Abi George;Park Seong Ok;Lee John Hwa;Chae Joon Seok;Cho Jeong Gon;Song Hee Jong
    • Journal of Microbiology
    • /
    • v.43 no.5
    • /
    • pp.430-436
    • /
    • 2005
  • In this study, the prevalence and quantity of a latent pseudorabies virus (PrV) infection in the nervous tissues of randomly selected pigs was determined via nested and real-time PCR. The nervous tissues, including the trigeminal ganglion (TG), olfactory bulb (OB), and brain stem (BS), were collected from the heads of 40 randomly selected pigs. The majority of the nervous tissues from the selected pigs evidenced a positively amplified band on nested PCR. In particular, nested PCR targeted to the PrV glycoprotein B (gB) gene yielded positive results in all of the BS samples. Nested PCR for either the gE or gG gene produced positive bands in a less number of nervous tissues ($57.5\%$ and $42.5\%$, respectively). Real-time PCR revealed that the examined tissues harbored large copy numbers of latent PrV DNA, ranging between $10^{0.1}\;and\;10^{7.2}(1-1.58{\times}10^7)$ copies per $1{\mu}g$ of genomic DNA. Real-time PCR targeted to the PrV gE gene exhibited an accumulated fluorescence of reporter dye at levels above threshold, thereby indicating a higher prevalence than was observed on the nested PCR ($100\%$ for BS, $92\%$ for OB, and $85\%$ for TG). These results indicate that a large number of farm-grown pigs are latently infected with a field PrV strain with a variety of copy numbers. This result is similar to what was found in association with the human herpes virus.

Analysis of Total Bacteria, Enteric Members of γ-proteobacteria and Microbial Communities in Seawater as Indirect Indicators for Quantifying Biofouling

  • Lee, Jin-Wook;Kim, Sung-Min;Jung, Ji-Yeon;Oh, Byung-Soo;Kim, In S.;Hong, Soon-Kang
    • Environmental Engineering Research
    • /
    • v.14 no.1
    • /
    • pp.19-25
    • /
    • 2009
  • In this study, total bacteria, enteric members of the $\gamma$-proteobacteria, and microbial communities in seawater were analyzed as indirect indicators for quantifying biofouling. Biomass in seawater can significantly affect feed water pretreatment and membrane biofouling of reverse osmosis desalination processes. The purpose of this paper is to investigate microbiological quantity and quality of seawater at the potential intake of a desalination plant. For this analysis, the total direct cell count (TDC) using 4'-6-diamidino-2-phenylindole (DAPI)-staining and DNA-based real-time PCR were used to quantify the total bacteria and relative content of enteric members of $\gamma$-proteobacteria in seawater, respectively. In addition, microbial communities were examined using 16S rRNA gene cloning and bacterial isolation to identify the most abundant bacteria for a further biofouling study. The experimental results of this study identified about $10^6$ cells/mL of (total) bacteria, $10^5$ 16S rRNA gene copies/mL of enteric $\gamma$-proteobacteria, and the presence of more than 20 groups of bacteria.

AN EXPERIMENTAL STUDY OF PLATELET-DERIVED GROWTH FACTOR ABOUT BONE FORMATION IN DENTAL IMPLANT (혈소판 유래 성장인자가 임플란트 주위 골형성에 미치는 효과에 대한 실험적 연구)

  • Ryu, Jae-Jun;Kwon, Jong-Jin;Kim, Han-Kyum
    • The Journal of Korean Academy of Prosthodontics
    • /
    • v.36 no.6
    • /
    • pp.816-831
    • /
    • 1998
  • Bony fixation of implants during the early phase of healing is important in order to get secondary stability of the implant assuring the success of the treatment. Because the successful placement of the implant is limited by the quality and quantity of bone, other agents which stimulate bone formation in the peri-implant spaces has been illustrated. Platelet-derived growth factor (PDGF) has been shown to regulate DNA and protein synthesis in bone cells in vitro and to interact synergistically to enhance soft tissue wound healing in vivo. The purpose of this study was to evaluate bone promotion around implants which were augmented with sagittal split osteotomy or autogenous veneer bone graft using the platelet derived growth factor(PDGF). After placement of newly designed twenty four screw-type implants, which were 12mm in length and 4mm in diameter in 6 dogs. $4{\mu}g$ of PDGF B/B was applied with surgicel carriers. The dogs were sacrificed at 3 days, 1, 2, 3, 6, and 12 weeks after implantation. Specimens were examined clinically, radiographically, histologically, and histomorphometrically. The results were as follows: 1. Clinically and radiologically, there was no significant difference in bone formation and healing pattern between experimental and control group. 2. In autogenous veneer bone graft group, bone formation was observed at 1st week in the experimental groups but 2nd week in the control groups. At 3rd week, the expeimental groups showed more bone formation comparing to the control groups. 3. In sagittal split osteotomy group, bone formation was observed at 1st week in both groups. But the experimental groups showed more bone formation comparing to the control groups after 2nd week. 4. The bone growth rate of experimental group was more rapid than that of control group. These results indicated that PDGF did not affect the initiation of new bone formation, but it accelerated the bone formation at the early period.

  • PDF

Relationship between Differential Gene Expression in Ovary and Heterosis of Egg Number Traits in a Chicken Diallel Cross

  • Wang, Hui;Sun, Dong-Xiao;Yu, Ying;Wang, Dong;Zhang, Yuan
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.18 no.6
    • /
    • pp.767-771
    • /
    • 2005
  • In order to understand the molecular basis of chicken heterosis in reproduction traits, mRNA differential display (DDRT-PCR) methods were used to analyze the differential gene expression of ovary tissue between hybrids and their parental lines in a 4${\times}$4 diallel cross, involving 4 chicken breeds, which were White Plymouth Rock (E), CAU Brown (D), Silkies (C) and White Leghorn (A). Total of 331 differential displayed cDNA bands from 1,161 were displayed in the 4${\times}$4 diallel cross combinations with 30 pairs of primers, which shows the differences of gene expression between hybrids and their parental lines were very obvious in quantity and quality. Seven types of differential expression patterns were found: Co-dominance expressed pattern (T1), under-expression of parental fragments in hybrids (T2), over-expression of parental fragments in hybrids (T3), hybrid-absence expressed pattern (T4), single parentspecific expressed pattern (T5), dominant expression fragments of single parent in hybrids (T6), hybrid-specific expressed pattern (T7). Correlation analysis indicated that there were significant correlations between the pattern of T3 and the heterosis percentage of egg number of 32-week and 42-week old chickens(p<0.01), while there were negative significant correlations between the pattern of T7 and the heterosis percentage of egg number of 32-week and 42 week-old birds (p<0.01).

Quantitative distribution of denitrifying bacteria with nirS and nirK in MLE and A/O process (MLE와 A/O 공정에서의 nirS 와 nirK 를 가진 탈질미생물의 정량적 분포)

  • Lim, Dong-Seok;Kim, Yun-Jung;Kim, Hyung-Gun;Park, Seung-Guk;Chung, Tai-Hak
    • Journal of Korean Society of Water and Wastewater
    • /
    • v.26 no.4
    • /
    • pp.591-598
    • /
    • 2012
  • Denitrification is an important biological mechanism in wastewater treatment process because this process is technically to remove nitrogen from water to air. There have been lots of study about denitrification engineering and molecular biological research about denitrifying bacteria, respectively. However, combination of these researches was unusual and rare. This study is about the correlation between quantity of denitrifying bacteria and denitrification potential, and consists of NUR batch test as analysis method of denitrification potential and quantitative molecular analysis for denitrifying bacteria. Three reactors (A/O, MLE and A/O of nitrogen deficiency) are operated to get activated sludge with various denitrification potential. All samples which were acquired from reactors were measured denitrification potential by NUR test and NUiR test. Also, Real-time PCR was conducted for quantification of denitrifying bacteria composition in activated sludge. The various denitrification potentials were measured in the reactors. The denitrifiaction potential was the highest in MLE process and the reactor of the nitrogen deficiency showed the lowest. Genomic DNA of activated sludge was obtained and consequently, real-time PCRuse the primer sets of nirK and nirS were conducted to quantify genes involving denitrification reductase production. As the result of real-time PCR, nirK gene showed more significant influence on denitrification potential comapred with nirS gene.