• Title/Summary/Keyword: serial dilution

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Characteristics on Equine Herpesvirus Type 3 from Korea (국내분리 말구진 원인병원체(Equine herpesvirus type 3)의 특징)

  • Yang, Jae-Hyuk;Lim, Yoon-Kyu
    • Journal of Life Science
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    • v.21 no.8
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    • pp.1156-1162
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    • 2011
  • Equine coital exanthema caused by equine herpesvirus type 3 (EHV-3) is a venereal disease which seriously drops horse reproduction rates. Here, we isolated EHV-3 from infected horses and investigated their biological characteristics. Initial cytopathic effects such as rounding of cells were detected 48 hours post infection of the virus into RK-13 cells. The infected cells were going to detach from the surface of culture flasks 72 hours post infection. The type of isolated viruses from swabbed samples was EHV-3 by PCR analysis. Glycoprotein G (gG) of isolated EHV-3 has a 99.25 percent similarity rate to that of EHV-3 334/74 control strain. The isolated EHV-3 was named Georo strain. Georo strain consisted of four major proteins including 145 kD, 60 kD, 45 kD and 40 kD, as shown by SDS-PAGE analysis. We hope the newly isolated Georo strain of EHV-3 can be used for studying various aspects of Korean equine coital exanthema.

Antimicrobial Activity of Elfvingia applanata Extract Alone and in Combination with Naringenin (Elfvingia applanata 엑스의 항균력 및 Naringenin과의 병용효과)

  • Cheon, Jun-Hee;Lee, Sun-Ok;Rym, Kyo-Hwan;Eo, Seong-Kug;Lee, Chong-Kil;Han, Seong-Sun
    • The Korean Journal of Mycology
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    • v.23 no.2 s.73
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    • pp.153-160
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    • 1995
  • As part of our search for less toxic antimicrobial agents from natural resources, the antimicrobial activity of Elfvingia applanata $(P_{ers.})\;K_{arst.}$ extract was examined alone and in combination with naringenin. EA, the aqueous extract from the carpophores of E. applanata, was lyophilized and a dark brownish powder was obtained. Antimicrobial activity of EA was tested in vitro against nineteen strains of bacteria and eleven strains of fungi by serial broth dilution method, and expressed by minimal inhibitory concentration (MIC). Among nineteen strains of bacteria tested, the antimicrobial activity of EA was the most potent against Proteus vulgaris showing MIC of 1.125 mg/ml. EA also inhibited the growth of the selected fungi at higher concentrations ranging from 7.5 mg/ml to 15.0 mg/ml. To investigate the effect of antimicrobial combinations of EA with naringenin, the fractional inhibitory concentration index (FICI) was determined by checkerboard assay for each strain. The antimicrobial combinations of EA with naringenin resulted in partial synergism against Staphylococcus aureus only, and showed additive effect in two strains including Klebsiella pneumoniae and Salmonella typhi. Antagonism was not found.

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Biological Control of Sclerotinia sclerotiorum in Lettuce Using Antagonistic Bacteria (길항세균을 이용한 상추 균핵병의 생물학적 방제)

  • Chon, Bong-Goan;Park, Suji;Kim, Jin-Won
    • Research in Plant Disease
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    • v.19 no.1
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    • pp.12-20
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    • 2013
  • To isolate antagonistic bacteria against sclerotinia rot of lettuce, caused by Sclerotinia sclerotiorum, soil samples were collected from the diseased greenhouse field in Namyangju city, Gyeong-gi province from 2007 to 2008. A total of 196 bacterial isolates were isolated using serial dilution method. In dual culture assay in vitro, 26 isolates showed more than 80% of inhibition rates of mycelial growth of S. sclerotiorum. Based on 16S rDNA sequence analysis, the 26 isolates were identified as Bacillus megaterium, B. cereus, B. subtilis, Arthrobacter nicotianae, A. ramosus, Pseudomonas filiscindens, Stenotrophomonas maltophilia, Brevibacterium frigoritolerans and Sphingobacterium faecium. The 26 isolates inhibited the mycelial growth of S. sclerotiorum up to 80% and the sclerotial germination 0-100%. In the greenhouse pot test of ten isolates conducted in summer, 2 isolates B. megaterium (DK6) and B. cereus (C210) showed control efficacy on sclerotia viability of S. sclerotiorum, 20% and 35%, respectively. In the greenhouse pot test in winter, the disease incidence of the control group was 80%, whereas those of 9 isolates among 26 were approximately 20%. From the result, the 9 isolates are expected as potentially antagonistic bacteria for biological control of sclerotinia rot of lettuce caused by S. sclerotiorum.

Phylogenetic diversity and UV resistance analysis of radiation-resistant bacteria isolated from the water in Han River (한강물로부터 분리된 방사선 내성 세균들의 계통학적 다양성 및 UV 내성 분석)

  • Lee, Jae-Jin;Joo, Eun Sun;Lee, Do Hee;Jung, Hee-Young;Kim, Myung Kyum
    • Korean Journal of Microbiology
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    • v.52 no.1
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    • pp.65-73
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    • 2016
  • The aim of this study was to investigate the UV-resistance of radiation-resistant bacteria isolated from the water of Han River, South Korea. The water sample was irradiated with 3 kGy gamma radiation prior to isolation. Radiation-resistant bacterial strains were isolated by standard serial dilution method on R2A and 1/10 diluted R2A agar. The resulting purely isolated 60 cultures of bacteria were analysed for UV resistance and used in further studies. Based on the comparative analyses of 16S rRNA gene sequences, the bacterial isolates were divided into 3 phyla (4 genera): the phylum Deinococcus-Thermus (the genus Deinococcus) was 61.7%, Bacteroidetes (Hymenobacter and Spirosoma) was 23.4%, and Firmicutes (Exiguobacterium) was 15%. The results suggested that twenty-nine isolates are candidates new species belonging to Deinococcus, Hymenobacter, and Spirosoma, or other new genera. Nine bacterial strains were selected among the novel candidates and the UV-resistance analysis was conducted. All the candidate bacterial strains showed high UV resistance, similar to that of D. radiodurans R1.

Antimicrobial Activities of Combined Extract of Aloe vera with Propolis against Oral Pathogens (알로에 베라 및 프로폴리스 혼합 추출물의 구강내 병원균에 대한 항균활성)

  • 임지영;문유선;정승희;이규임;유수연;박원봉;심창섭
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.5
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    • pp.899-904
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    • 2002
  • Aloe and propolis are extensively used in folk medicine. Ethanol extracts of Aloe vera (AE), ethanol extract of propolis (PE) and waxfree extract of propolis (PW) were prepared to test antimicrobial activities against five oral microorganisms (Streptococcus mutans, Enterococcus faecalis, Enteococcus hirae, Escherichia coli, Candida albicans). Antimicrobial activities were tested by serial broth dilution method and expressed by minimal inhibitory concentration (MIC). The AE showed relatively weak antimicrobial activities, while both of PE and PW greatly inhibited all microorganisms tested. To investigate the antimicrobial effects of the combined extracts of aloe with propolis, the fractional inhibitory concentration index (FICI) was determined by checkerboard assay for each strain. The combination of AE with PE or PW resulted in Synergistic effect against oral microorganisms tested (FICI=0.375) except Escherichia coli (FICI=1.0 for PE, FICI=0.75 for PW).

Fusion Analytical Sensitivity of Rapid Influenza Antigen Limit of Detection Tests for Human Influenza virus (인플루엔자 바이러스에 대한 신속 항원 검출 검사 검출한계의 융합적 분석)

  • Song, Chang-Sub;Sung, Hyun-Ho;Kim, Jung-Hyun;Kim, Dae-Eun;Park, Chang-Eun;Yoon, Joong-Soo
    • Journal of the Korea Convergence Society
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    • v.9 no.3
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    • pp.165-171
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    • 2018
  • In this study, to analyze the detection of limit for sensitivity of the influenza rapid antigen test kit, the positive detection of limits were analyzed by serial dilution of influenza virus A and B type for five influenza rapid antigen test kits in Korea. As a result of analysis, visual measurement of type A were up to 1:8192 for the Wellsbio product and up to 1:4096 for the II product, up to 1:512 for the I and III products, and only 1:128 for the IV product, and type B were positive for up to 1:8192 for the Wellsbio product, up to 1: 4096 for the II product and up to 1:1024 for the I, III and IV products. For instrument readings with the same specimen, both A and B types were found to be positive for up to 1:8192 for the Wellsbio product, up to 1: 4096 for the II product, and up to 1:2048 for the I product. The sensitivity of the rapid antigen test for influenza differs greatly depending on the sampling area of the patient, infection period, specimen volume, etc. Therefore, it is necessary to observe exactly the collection timing and method of the specimen. And it is necessary further study to improve the sensitivity for influenza rapid antigen test.

Effects of Swim Training on Tuberculosis infection in the Mouse Model at Different Temperature (온도 차이에 따른 수영훈련이 결핵균 감염정도에 미치는 영향)

  • Kwak Yi-Sub
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.652-656
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    • 2005
  • Tuberculosis is the leading infectious disease in the world. It is urgent to develop new vaccine and treating drugs. Besides vaccines, we want to know the effects of regular swim training on TB infection in the mouse model. This study was designed to examine the effects of regular swim training on lung and spleen TB counts and $INF-\gamma$ activity in the trained mice at different temperature. The trained mice underwent a 10-wk endurance swim training (5 times/wk) in water at $29\~33^{\circ}C$ (WWG) and $21\~23^{\circ}C$(CWG) for 60 min. And they were divided into 3 groups according to the regular swim training (CG; control, WWG; warm water group, and CWG; cold water group). Mice were challenged by aerosol infection with M. tuberculosis H37Rv using an inhalation device (Glas-Col, Terre Haute, Ind.) calibrated to deliver bacteria into lungs. Three weeks after immunization, the mice were challenged. Four weeks after challenge, the mice were sacrificed and the numbers of viable bacteria in lung and spleen were determined by plating serial dilution of whole organ homogenates on nutrient Middlebrook 7H11 agar (Difco, Detroit, MI). Colonies were counted after four weeks incubation at $37^{\circ}C$. All data were expressed as mean, standard deviation by using SPSS package program (win 10.0). The result through the statistical analysis of this data were summarized as follows; In the weight changes, there were significant differences among CG, WWG, and CWG following the swim training at different temperature, and CWG was the lowest. In the change of $INF-\gamma$ following the swim training, there were significant differences (p<.05) among CG, WWG, and CWG after stimulated with media and CFP. In MTB counts, there were significant differences (p<.05) between CG and WWG in the lung. And also there were significant differences (p<.05) among CG, WWG, and CWG. These results suggest that regular swim training suppress Th1 immune response caused by decreased $INF-\gamma$ level in the WWG, Also For the WWG, highly increased level of TB counts appear in the lung and spleen compare to CG.

Species-specific Marker Development for Environmental DNA Assay of Endangered Bull-head Torrent Catfish, Liobagrus obesus (멸종위기어류 퉁사리의 환경 DNA 분석을 위한 종 특이 마커 개발)

  • Yun, Bong Han;Kim, Yong Hwi;Sung, Mu Sung;Han, Ho-Seop;Han, Jeong-Ho;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.34 no.3
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    • pp.208-217
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    • 2022
  • We wanted to develop a real-time PCR assay capable of detecting Liobagrus obesus in environmental DNA (eDNA) extracted from freshwater samples using a pair of species-specific primers and probe for the endangered fish, L. obesus. The species-specific primers and probe were designed in consideration of single nucleotide polymorphisms between 65 species of freshwater fish living in the Republic of Korea within the cytochrome b (cytb) gene of mitochondrial DNA. The species-specific primers and probe, in the real-time PCR assay, showed high specificity as only the L. obesus genomic DNA (gDNA) was found to be positive in the specificity verification using 65 species gDNA of freshwater fish in the Republic of Korea. In addition, in the detection limit analysis using the serial dilution concentrations of L. obesus gDNA, it was found that it was possible to detect up to 0.2 pg, showing high sensitivity. Afterwards, using the species-specific primers and probe, real-time PCR assay was performed on freshwater samples obtained from 8 stations in the mid-upper basin of Geum River. As a result, the cytb gene of L. obesus was detected in total 5 stations including all 3 stations where this species was collected at the time of field survey. Therefore, the species-specific primers and probe developed in present study, and the real-time PCR assay using them, can accurately detect the cytb gene of L. obesus from eDNA samples, which can be utilized to monitor the existing habitats of this species and to discover potential new habitats.

A STUDY ON THE DISTRIBUTION OF CALCITONIN GENE-RELATED PEPTIDE CONTAINING NERVE FIBERS IN RAT PULP FOLLOWING DENTINAL INJURY (상아질 손상 후 흰쥐 대구치 치수의 calcitonin gene-related peptide(CGRP) 함유 신경섬유 분포에 관한 연구)

  • Moon, Joo-Hoon;Park, Sang-Jin;Min, Byung-Soon;Choi, Ho-Young;Cho, Gi-Woon
    • Restorative Dentistry and Endodontics
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    • v.24 no.1
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    • pp.100-115
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    • 1999
  • The purpose of this study was to investigate the distribution of calcitonin gene-related peptide containing nerve fibers in rat pulp after dentinl injury by means of immunohistochemistry and confocal laser scanning microscope. The Spague-Dawley rats weighing about 250-300gm were used. The animals were devided into normal control and experimental groups. Experimental animals were sacrified 1, 2, 4, 7, 10, 21days after dentinal injury (dentin cutting, and then acid etching with 35% phosphoric acid) on the maxillary molar teeth. The maxillary teeth and alveolar bone were removed and immersed in the 4% paraformaldehyde in 0.1M phosphate buffer (pH 7.4), then were decalcified with 15% formic acid for 10 days. Serial frozen $50{\mu}m$ thick sections were cut on a cryostat. The rabbit CGRP antibody was used as a primary antibody with a dilution of 1:2000 in 0.01M PB. The sections were incubated for 48 hours at $4^{\circ}C$, and placed into biotinylated antirabbit Ig G as a secondary anti body with dilution of 1:200 in 0.01M PB and incubated in ABC(avidin-biotin complex). The peroxidase reaction was visualized by incubating the sections in 0.05% 3,3 diaminobenzidine tetrahydrochloride containing 0.02% $H_2O_2$. For the confocal laser scanning microscopic examination, Primary antibody reaction was same as immunoperoxidase stainning, but fluorescein isothiocyanate(FITC)-conjugate antirabbit IgG as a secondary antibody was used. The confocal laser scanning microscope was used for the examination. A series of images of optical sections was collected with a 20x objective at $3{\mu}m$ intervals throughout the depth of specimen. FITC fluerescence was registrated through a 488nm and 568nm excitation filter, and images were saved on optical disk. The stereoscopic images and three dimentionnal images were reconstructed by computer software, and then were analyzed. The results were as follows : 1. In normal control group, CGRP containing nerve fibers were coursed through the root with very little branching, and then formed a dense network of terminals in coronal pulp. 2. A slight increase in CGRP containing nerve fibers at 1 and 2day postinjury was noted subjacent to the injury site. In the 4day group, there were an extensive increase in the number of reactive fibers, followed by a partial return toward normal levels at 7~10 day postinjury, and return by 21days. 3. The sprouting of the CGRP containing nerve fibers was evident within 2day after dentinal injury, and by 4days there was a maximal increased, but was decreased at 7days and returned to normal 10~21 day postinjury. 4. In confocal laser scanning microscopic exammination, the distinct distribution pattern and sprouting reaction of CGRP containing nerve fibers were observed in stereoscopic images and three dimentional images. These results suggest that CGRP containing nerve fiber can be important role in the response to dental injury and pain regulation.

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Polishing characteristics of polyetherketoneketone on Candida albicans adhesion (Polyetherketoneketone의 연마 특성이 Candida albicans의 부착에 미치는 영향)

  • Kim, Hyunyoung;Lee, Jonghyuk;Lee, Sung-Hoon;Baek, Dongheon
    • The Journal of Korean Academy of Prosthodontics
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    • v.58 no.3
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    • pp.207-216
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    • 2020
  • Purpose: To compare the polishing characteristics and their influence on Candida albicans adhesion to the recently introduced polyetherketoneketone (PEKK) and the conventional polymethylmethacrylate (PMMA) denture resin material. Materials and methods: Specimens from PEKK (Group E) and PMMA (Group M) were made in dimensions of 8 mm in diameter and 2 mm in thickness. The specimens were further divided into sub-groups according to the extent of polishing (ER, MR: rough; EP, MP: polished, N = 12 each). The specimens were polished using polishing machine and SiC foil. ER and MR group specimens were polished with 600 grit SiC foil only. EP and MP groups were further polished with 800, 1,000, 1,200 grit SiC foils sequentially. To measure the surface roughness values (Sa) of specimens, atomic force microscope (AFM) was used and scanning electron microscope (SEM) observation under 1,000, and 20,000 magnifications was performed to investigate surface topography. The polished specimens were soaked in C. albicans suspension for 2 hours with shaking to promote adhesion. The attached C. albicans were detached from the surface with 10 times of pipetting. The suspension of detached C. albicans was performed by serial dilution to 103 times, and the diluted suspensions were inoculated on Sabouraud dextrose agar plates using spread plate method. After incubating the plate for 48 hours, colony forming unit (CFU)/plate of C. albicans was counted. Statistical analysis was performed using one-way ANOVA and Tukey HSD test to confirm significant difference between the groups (α=.05). Results: Average Sa value was significantly higher in MR group compared to other groups (P<.05), meaning that additional polishing steps reduced surface roughness effectively only in the PMMA specimens. There was no significant difference in Sa values between MP and EP groups. In SEM images, PEKK specimens showed numerous spikes of abraded material protruding from the surface and this phenomenon was more significant in EP group. The mean CFU/plate value was the highest in EP group and this was significant when it was compared to MP group (P<.05) which was the lowest. Conclusion: Polishing PEKK using serial SiC abrasive foil may result in higher adhesion of C. albicans. In clinic, this should be considered carefully.