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Genotype Frequency of Human Papillomavirus Determined by PCR and DNA Sequencing in Korean Women

  • Lee, Kyung-Ok;Seong, Hye-Soon;Chung, Soo-Jin;Jung, Na-Young;Lee, Hye-Jung;Kim, Kyung-Tae
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.2
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    • pp.99-104
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    • 2006
  • Human Papilloma viruses (HPVs) are etiological agents for cervical cancer and are classified into low- and high-risk categories. The aim of this study was to determine the frequency of the HPV genotype in the HPV screening test of Korean women using PCR-direct sequencing. Consensus primers of L1 legion were used for the amplification of HPV DNA and the PCR products (450 bps) obtained were analyzed by automatic sequencing. Sequences were compared with those in GenBank by using the BLAST program. Cervical swab samples of 3,978 women (20-73 years) were tested and the average age was 37.6 years. In this study, 1,174 samples were HPV positive out of 3,978 cervical swab samples screened (29.5%) and 136 samples (11.6%) showed a double infection. A total of 1,310 HPV genotypes were analyzed. The HPV positive rate was the lowest in the 20 years group (69.5%) and most of the samples of the > 60 years group were found HPV positive. Among thirty seven different HPV types identified by sequencing, 21 were HPV high risk types and 16 HPV low risk types were 69.8% (914/1,310) and 26.0% (340/1,310), respectively. In HPV high-risk types, 16 (13.21%), was the most frequently found. HPV 53 (9.62%) and 58 (9.24%) were also frequently found. This group was followed by HPV types 70 (5.50%), 33 (4.73%), 66 (4.20%), 18 (4.05%), 52 (4.05%), 31 (3.97%) and 56 (3.51%) in descending order of frequency. Among HPV low-risk types, 62 (4.20%), 6 (3.59%), 81 (3.59%), 84 (3.51%), and 11 (2.6%) were frequently found. In conclusion, PCR-direct sequencing could be used for quick and reliable typing of known and novel HPVs from clinical specimens. This data could be useful for epidemiological study of HPV and it also allows type-specific follow-up of women who have been treated for cervical intraepithelial neoplasia.

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Establishment of Bovine Ovum Bank : I. Full Term Development of Vitrified In Vitro Matured Hanwoo (Korean Cattle) Oocytes by Minimum Volume Cooling (UC) Method (소 난자 은행 설립 : I. MVC 방법으로 초자화 동결된 한우 미성숙 난자의 개체 발생능 조사)

  • Kim, E.Y.;Kim, D.I.;Rhee, M.G.;Weon, Y.S.;Nam, H.K.;Lee, K.S.;Park, S.Y.;Park, E.M.;Yoon, J.Y.;Heo, Y.T.;Cho, H.J.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Korean Journal of Animal Reproduction
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    • v.25 no.1
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    • pp.1-7
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    • 2001
  • This study was to test whether in vitro matured Hanwoo oocytes can be successfully cryopreserved by a new vitrification procedure using MVC method. For the vitrification, oocytes were pretreated in 10% ethylene glycol (EG10) for 5~10 min, exposed in EG30 for 30 sec, each oocyte was individually put on the inner wall of 0.25 $m\ell$ straw, and then straws were directly plunged into L$N_2$. Thawing was taken by 4-step procedures 〔1.0 M sucrose (MS), 0.5 MS, 0.25 MS, and 0.125 MS〕 at 37$^{\circ}C$. In vitro developmental capacity (survival, cleavage ($\geq$2-cell) and blastocyst rates) in vitrified group was no significant difference compared to that in other treatment groups (exposed; 100.0, 74.4, 32.3% and control; 100.0, 78.3, 36.3%): high mean percentage of oocytes (91.2%) was survived, 69.4% of them were cleaved and 27.9% of cleaved embryos were developed to blastocyst. Especially, after transfer of in vitro developed embryos in vitrified group, four of six recipient animals were pregnant and three of them were ongoing-pregnant by manual palpation at 250 days after transfer. This result demonstrates that MVC method is very appropriate freezing method for the Hanwoo in vitro matured oocytes and that ovum bank can be maintained efficiently by MVC cryopreservation method.

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A Study on The Measurement of Cerebral Cortical Thickness in Patients with Mood Disorders (기분장애 환자의 대뇌 피질 두께 측정에 관한 연구)

  • Do-Hun Kim;Hyo-Young Lee
    • Journal of the Korean Society of Radiology
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    • v.18 no.2
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    • pp.73-81
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    • 2024
  • This study compared the cortical thickness of patients with mood disorders and a control group to assess structural abnormalities. A retrospective study was conducted from September 2020 to August 2022 at the Department of Psychiatry, P Hospital in Yangsan, Gyeongsangnam-do. The study included 44 individuals diagnosed with mood disorders and 59 healthy individuals without any pathological lesions. The 3D-T1 MPRAGE images obtained from magnetic resonance imaging examinations were utilized, and FreeSurfer software was employed to measure cortical thickness. Statistical analysis involved independent samples t-tests to measure the differences in means between the two groups, and Cohen's d test was used to compare the effect sizes of the differences. Furthermore, the correlation between the measured average cortical thickness and Positive and Negative Syndrome Scale scores was analyzed. The research results revealed that patients with mood disorders exhibited decreased cortical thickness compared to the normal control group in both superior frontal regions, both rostral middle frontal regions, both caudal middle frontal regions, both pars opercularis, pars orbitals, pars triangularis regions, both superior temporal regions, both inferior temporal regions, both lateral orbitofrontal regions, both medial orbitofrontal regions, both fusiform regions, both posterior cingulate regions, both isthmus cingulate regions, both superior parietal regions, both inferior parietal regions, both supramarginal regions, left postcentral region, right bank of the superior temporal sulcus region, right middle temporal region, right rostral anterior cingulate region, and right insula region. Among them, regions that showed differences with effect sizes of 0.8 or higher were left fusiform (d=0.82), pars opercularis (d=0.94), superior frontal (d=0.88), right lateral orbitofrontal (d=0.85), and pars orbitalis (d=0.89). Additionally, there was a weak negative correlation between PANSS scores and average cortical thickness in both the left hemisphere (r=-0.234) and right hemisphere (r=-0.230). These findings are expected to be helpful in identifying areas of cortical thickness reduction in patients with mood disorders compared to healthy individuals and understanding the relationship between symptom severity and cortical thickness changes.

Cloning and Expression of the Cathepsin F-like Cysteine Protease Gene in Escherichia coli and Its Characterization

  • Joo, Han-Seung;Koo, Kwang-Bon;Park, Kyun-In;Bae, Song-Hwan;Yun, Jong-Won;Chang, Chung-Soon;Choi, Jang-Won
    • Journal of Microbiology
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    • v.45 no.2
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    • pp.158-167
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    • 2007
  • In this study, we have cloned a novel cDNA encoding for a papain-family cysteine protease from the Uni-ZAP XR cDNA library of the polychaete, Periserrula leucophryna. This gene was expressed in Escherichia coli using the T7 promoter system, and the protease was characterized after partial purification. First, the partial DNA fragment (498 bp) was amplified from the total RNA via RT-PCR using degenerated primers derived from the conserved region of cysteine protease. The full-length cDNA of cysteine protease (PLCP) was prepared via the screening of the Uni-ZAP XR cDNA library using the $^{32}P-labeled$ partial DNA fragment. As a result, the PLCP gene was determined to consist of a 2591 bp nucleotide sequence (CDS: 173-1024 bp) which encodes for a 283-amino acid polypeptide, which is itself composed of an 59-residue signal sequence, a 6-residue propeptide, a 218-residue mature protein, and a long 3'-noncoding region encompassing 1564 bp. The predicted molecular weights of the preproprotein and the mature protein were calculated as 31.8 kDa and 25 kDa, respectively. The results of sequence analysis and alignment revealed a significant degree of sequence similarity with other eukaryotic cysteine proteases, including the conserved catalytic triad of the $Cys^{90},\;His^{226},\;and\;Asn^{250}$ residues which characterize the C1 family of papain-like cysteine protease. The nucleotide and amino acid sequences of the novel gene were deposited into the GenBank database under the accession numbers, AY390282 and AAR27011, respectively. The results of Northern blot analysis revealed the 2.5 kb size of the transcript and ubiquitous expression throughout the entirety of the body, head, gut, and skin, which suggested that the PLCP may be grouped within the cathepsin F-like proteases. The region encoding for the mature form of the protease was then subcloned into the pT7-7 expression vector following PCR amplification using the designed primers, including the initiation and termination codons. The recombinant cysteine proteases were generated in a range of 6.3 % to 12.5 % of the total cell proteins in the E. coli BL21(DE3) strain for 8 transformants. The results of SDS-PAGE and Western blot analysis indicated that a cysteine protease of approximately 25 kDa (mature form) was generated. The optimal pH and temperature of the enzyme were determined to be approximately 9.5 and $35^{\circ}C$, respectively, thereby indicating that the cysteine protease is a member of the alkaline protease group. The evaluation of substrate specificity indicated that the purified protease was more active towards Arg-X or Lys-X and did not efficiently cleave the substrates with non-polar amino acids at the P1 site. The PLCP evidenced fibrinolytic activity on the plasminogen-free fibrin plate test.

Strength Characteristics of the Soil Mixed with a Natural Stabilizer (친환경 토양안정재를 혼합한 지반의 강도특성)

  • Kwon, Youngcheul;Oh, Sewook
    • Journal of the Korean GEO-environmental Society
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    • v.13 no.1
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    • pp.45-51
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    • 2012
  • This article aims to find method to mix a harmless hardening agent and soil generated during construction to make paving materials. The main purpose of this research is to get rid of the harmfulness(Chromium (VI), etc.) of cement which has been generally and frequently used as a hardening agent and strengthen it so that it can be used for the general foundation solidification and stabilization of civil engineering/construction structures such as dredging soil treatment, marine structure foundation treatment, surface soil stabilization, and river bank erosion prevention. NSS(Natural Stabilizer Soil) used for this study takes as its chief ingredient the mixture of lime and staple fibers extracted from natural fibers. It increases the shearing strength of soil that it improves the support and durability of the foundation and prevents flooding and frost as well. The pH measured to know its eco-friendliness was 6.67~7.15, and according to the migration testing, only Pb and CN were lower than the standards, so it can be said that NSS has almost no harmful components in it. According to the result of uniaxial strength testing, when the mixture ratio of weathered soil to NSS was 6%, about 1,850kpa strength was expressed. And according to the result of CBR. testing to figure out its appropriateness as a paving material, the CBR of the foundation was 4%~6%. But when the mixture ratio of NSS is over 6%, the water immersion CBR. is over 100%; thus, it is expected that it will show great utility as a paving material.

Specific Detection of Serratia marcescens Based on a PCR Assay and Antimicrobial Susceptibility of S. marcescens Isolated from Boar Semen (Serratia marcescens 검출을 위한 PCR 기법 개발 및 돼지정액 유래균주에 대한 항생제 감수성 양상)

  • Jung, Ji-A;Kim, Aeran;Seo, Byoung Joo;Jung, Suk Chan;Kim, In Cheul;Chung, Ki Hwa;Jung, Byeong Yeal
    • Journal of Life Science
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    • v.23 no.9
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    • pp.1133-1139
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    • 2013
  • During the collection of boar semen, bacterial contamination usually occurs. The contamination has deleterious effects both on semen quality and on sow fertility. The majority of contaminants are gram-negative bacteria, especially Serratia marcescens. In this study, we developed a PCR assay for the identification of S. marcescens targeting the luxS gene (GenBank no. EF164926). S. marcescens yielded a specific 306 bp PCR product. However, no amplification was observed in the other strains tested. The detection limit of PCR was $50pg/{\mu}l$ of template DNA of S. marcescens. The antimicrobial susceptibility patterns of S. marcescens isolated from boar semen were tested using the disk diffusion method. Gentamicin, ceftiofur, florfenicol, and neomycin showed high sensitivity in this test. The minimum inhibitory concentration (MIC) was also determined by the broth microdilution method. The $MIC_{90}$ values of ceftiofur, enrofloxacin, gentamicin, and neomycin were 8, 8, 8, and $16{\mu}g/ml$, respectively. These results indicate that PCR amplification of the luxS gene is a reliable and effective method for the identification of S. marcescens and that ceftiofur, enrofloxacin, gentamicin, and neomycin are effective semen extenders for controlling S. marcescens.

Stability Analysis of Low Flow Revetments on External Forces (저수호안에 작용하는 외력에 의한 안정성분석)

  • Kim, Chul;Park, Nam-Hee;Kim, Dae-Young;Kim, Yun-Hwan
    • Journal of the Korean Society of Hazard Mitigation
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    • v.8 no.5
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    • pp.147-153
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    • 2008
  • Tractive forces by flowing water and ship generated waves are items that affect the stability of the low flow revetments among various external forces exerting on those revetments. Bank revetment stability by these external forces is analysed in this study. The study area is the section of the test construction area changing the artificial revetments to ecological revetments in Han river. Tractive forces are computed using the calculated flow velocity using RMA-2 model. The stability is analyzed comparing the calculated tractive forces with permissible tractive forces of the revetments in the study area. The calculated tractive forces at section number 93 is higher than permissible tractive forces in that section, so the section is estimated hydraulically unstable. The calculated tractive forces for the storm of 10th August 2007 are small compared to the permissible tractive forces in all sections. The sections are considered to be hydraulically stable, but have been eroded in some parts. The reason for the erosion is considered to have insufficient time for the plants taking root, and be exerted composite forces such as forces by ship generated waves. Ship generated waves by the excursion boats and small boats called river taxi was calculated. Wave forces by these calculated waves are computed and compared with the supporting forces of the revetment material. The external forces exerted by the ships in Han River on the revetments is very little compared with the permissible supporting forces of the revetments, so the revetments are estimated hydraulically stable. But considering the composite forces are exerted simultaneously, the stability consideration should include these composite forces.

An Experimental Study of the Healing Process on Several Heterogenous Bone Grafts in Rabbit Mandible;Histologic and Biometric study (가토(家兎) 상악골(上顎骨) 결손부(缺損部)에 수종(數種)의 이종골(異種骨) 이식후(移植後) 치유과정(治癒過程)에 관(關)한 연구(硏究))

  • Oh, Hee-Kyun;Ryu, Sun-Youl
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.11 no.1
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    • pp.117-129
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    • 1989
  • The commercial availability of processed heterogenous bone has provided the surgeons with almost unlimited supply, avoidance of additional operation and prevention of the postoperative complications. In addition to these merits, unnecessary bone bank, easy availibility and storage have been achieved. The purpose of this study was to compare and examine the healing capacity of Kiel bone, Pyrost and Osteovit which used as the processed heterografts for the reconstruction of bony defect. Twenty rabbits weighing about 1.7-2.0 Kg were selected and divided into two groups. In experimental group A, the left mandibular defect was allowed to fill with blood, and the right defect was filled with Kiel bone. In experimental group B, the left defect was grafted with Pyrost, and the right with Osteovit. The experimental animals were sacrified after 1, 2, 4 and 8 weeks and the grafted site was studied histologically. To evaluate the strength of healed bone, 2 rabbits from each experimental group and a nonoperated control were sacrified at the 6th week after implantation and used for biometric testing on universal testing machine. The results obtained were as follows : 1. It was considered that these heterogenous bone grafts has feeble or absent immunogenicity since all of them appeared to evoke little inflammatory or forign body reaction. 2. In all experimental groups, new bone formation began from the adjacent region of host bone and extended progressively into the defect sites. New bone was partly formed within the intertrabecular space of the implant and gradually united with the bone that formed at the margin of the host bone. 3. With Pyrost bone formation was rapid and prominent comparing with other graft materials. 4. Osteovit was begun to be absorbed from 2 weeks, and Kiel bone from 4 weeks, however Pyrost was remained to be intact until the end of 8 weeks. 5. As the results of tensile test, the mean values of maximum tensile stress were 1.11${\uparrow}$ $Kgf/mm^{2}$ in Pyrost implanted specimens, 0.85 $Kgf/mm^{2}$ in Osteovit, 0.42 $Kgf/mm^{2}$ in Kiel bone, 0.66 $Kgf/mm^{2}$ in blood filled specimens and 1..13 $Kgf/mm^{2}$ in control. These results indicate that heterogenous bones grafted have little antigenicity to the host tissue, and that they mediate effectively osteoconduction by providing the scaffold for the bone formation. Pyrost and Osteovit appeared to be suitable for the clinical use.

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A Study on Development of Evaluation Method on Riverine Ecobelt (수변 생태벨트 평가방법 개발에 관한 연구)

  • Cho, Yong-Hyeon;Choi, Dae-Hee
    • Journal of the Korean Institute of Landscape Architecture
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    • v.42 no.1
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    • pp.123-132
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    • 2014
  • This study aims to develop the diagnostic evaluation method of the riverine ecobelt for construction, conservation, and maintenance of the riverine ecobelt. The value indices in the proposed evaluation method are composed of total 5 fields and 19 elements. The 5 fields are flood control, environmental function, growth of plants, ecobelt function, and restoration potential. Flood control field is composed of total 3 elements such as length, width, and density of green area. Environmental function field is composed of 4 elements such as park use, landscape boundary and edge, microclimate control, non-point pollution control. Growth of plants field is composed of 6 elements such as species composition, forest height, stratum structure, vine plants, plant vitality, and succession of plants. Ecobelt function field is composed of 4 elements such as longitudinal connectivity, lateral connectivity, in-stream forest or habitat, roads on bank top. Restoration potential field is composed of 2 elements such as landform and land use of the immediate vicinity. The score system ranging 1~4 was adopted. The weighting parameters of elements were unified with each other. The final grade system ranging 1~5(1: very good~5: very bad) was adopted, and the final grade was evaluated by the mean values of each field. According to the test application of the diagnostic evaluation method of the riverine ecobelt, the final grades showed effectively the real condition of each site.

Stem Rot on Ligularia fischeri Caused by Sclerotium rolfsii in Korea (Sclerotium rolfsii에 의한 곰취 흰비단병)

  • Moon, Youn-Gi;Kim, Se-Won;Choi, Jun-Keun;Kwon, Soon-Bae;Shim, Hong-Sik;Ju, Ho-Jong;Choi, In-Young
    • Research in Plant Disease
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    • v.21 no.1
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    • pp.36-39
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    • 2015
  • In June 2012 and 2013, a destructive stem rot symptoms of Ligularia fischeri occurred sporadically in Hoengseong-gun and Pyeongchang-gun Gangwon-do, Korea. The typical symptom included water-soaking on the main stem, rotting, wilting and blighting, which eventually leads to death of the plant. White mycelial mats were spread over lesions and brown sclerotia were formed on stems and near soil surface. The sclerotia were white to brown, spherical or irregular, 1-3 mm in size on potato dextrose agar (PDA), The optimum temperature range of hyphal growth was $25-30^{\circ}C$ and the hyphal diameter was $4-10{\mu}m$. The typical clamp connections were observed in the hyphae of the fungus grown on PDA. The resulting sequence of 695 bp was deposited in GenBank. A BLAST search revealed that sequences of the this isolates showed >99% identity with those of Sclerotium rolfsii. On the basis of the morphological characteristics and phylogenetic analyses of molecular markers ITS rDNA, the fungi were identified as S. rolfsii. A pathogenicity test was carried out to fulfill Koch's postulates. To our knowledge, this is the first report of S. rolfsii on Ligularia fischeri in Korea.