• The Journal of the Korean Society for Microbiology
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• v.34 no.5
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• pp.453-459
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• 1999

Astrovirus is frequently associated with diarrhea in children. It can not be readily isolated by cell culture, and an electronmicroscope is usually used for detection of this agent. Recently in 1995 a combined method of reverse transcription-polymerase chain reaction (RT-PCR) was designed for easier detection of astrovirus, which is based on the conserved sequence in 3'-end of genomes of the 7 known serotypes of human astrovirus. As of yet there has not been any report of astrovirus data in Korea using the RT-PCR methods. The purpose of this study was to detect astrovirus incidence, severity of symptoms, seasonal variation and co infection rate with rotavirus in Korean children inpatients with diarrhea. Fecal specimens from 61 young children hospitalized with gasteroenteritis Korea from Jan. 1996 through Mar. 1997. They were examined for astroviurs infection by RT-PCR method. Results are as follows:1. Astrovirus was detected at 9.8% (6/61) from fecal specimens of children with severe diarrhea by EIA using monoclonal antibody coated plates. 2. Astorvirus was detected at 29.5% (18/61) from fecal specimens of children with severe diarrhea by RT-PCR. 3. The age of the 18 children affected by astrovirus ranged from 2 monthes to 7 years with mean of 3.0 years. 4. Mean hospital stay of the 18 children was 6.1 days. 5. Five (27.8%) astrovirus RT-PCR positive strains were confirmed in November and in December, respectively out of 18 specimens in total. 6. Astrovirus coinfection with rotavirus type G1 was confirmed in 15/16 specimens (93.8%), and with type G2 was in 1/16 specimens (6.3%).

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.4 no.4
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• pp.363-370
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• 1999

When the first red tide alert by Cochlodinium polykrikoides was alarmed around the Oenarodo coast on Aug. 27, 1997, there co-occurred two chain-forming naked dinoflagellates which were different sized but looked fairly similar. The analyses of 24S rRNA sequences of these species showed that their gene sequences had only 74.9% identity. This low value implies that they are quite different species. After isolation and cultivation of each species, the morphological characteristics were observed. This revealed that the larger species ranging from 20 to 35 ${\mu}m$ was the well known, Cochlodinium polykrikoides and the smaller one ranging from 12 to 25 ${\mu}m$ was Gyrodinium impudicum which had not been reported in Korea. As their 24S rRNA sequences had not been analysed yet, we deposited the sequences in Genbank. At that time of the investigation. the red tide was caused by G. impudicum of which maximum cell counts reached up to 30,000 cells $ml^{-1}$. In this study we describe the morphological characteristics and the behavioral patterns of each species which can be easily observed with light microscope or stereomicroscope. In addition, their morphology transformed by the fixation with Lugol's solution are also characterized. which can help to discriminate each one in the fixed sample.

• Research in Plant Disease
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• v.26 no.3
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• pp.144-158
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• 2020

Quantitative reverse transcription PCR is used for gene expression analysis as the accurate and sensitive method. To analyze quantification of gene expression changes in apple plants, 10 housekeeping genes (ACT, CKL, EF-1α, GAPDH, MDH, PDI, THFs, UBC, UBC10, and WD40) were evaluated for their stability of expression during infection by Apple stem grooving virus (ASGV) or in cold-stress apple plant buds. Five reference-gene validation programs were used to establish the order of the most stable genes for ASGV as CKL>THFs>GAPDH>ACT, and the least stable genes WD40CKL>UBC10, and the least stable genes were ACT

• Pediatric Infection and Vaccine
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• v.23 no.2
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• pp.149-154
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• 2016

Although an association of Kawasaki disease (KD) with infectious agents has been suggested, none have been proven to cause KD. In this case study, we present a case of KD with concurrent onset of influenza and Mycoplasma pneumoniae (MP) infections. A 27-month-old boy presented with prolonged fever, cough, and rhinorrhea. During the initial testing, influenza A infection was identified, and he was treated with oseltamivir. Despite the antiviral therapy, the fever persisted, and he had cervical lymph node enlargement, bilateral conjunctival injection, fissured red lips, strawberry tongue, and erythematous skin lesions on the Bacillus Calmette-$Gu{\acute{e}}rin$ vaccination site. Thus, the patient was diagnosed with KD and was treated with intravenous immunoglobulin (IVIG). The result of the initial antimycoplasma immunoglobulin M (IgM) antibody testing and was positive, and an increased IgM titer from baseline was found in a repeat test. We reviewed the hypotheses on pathogens known to be associated with KD and the etiology of KD. Based on our findings, we suspect that symptoms of KD and coronary artery lesions can occur from various infections besides those caused by Mycoplasma species and influenza viruses.

• The Korean Journal of Applied Statistics
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• v.21 no.1
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• pp.81-94
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• 2008

Family-based tests such as the transmission and disequilibrium tests(TDT) have proved to be powerful tools in the search for disease genes. Unlike case-control studies, the tests are not affected by population admixture, which can lead to spurious association of multiple highly linked makers with disease-susceptible genes. Those tests have largely required knowledge of parental marker genotypes. However, parental data are often not available for late-onset diseases. In this article we propose sib-TDTs that overcome this problem by use of marker data from unaffected sib(s) instead of parents. To do this end, we fist defined a Mantel-Haenszel-type statistic for each haplotype and then proposed two tests based on this statistic. Simulation studies suggest that the proposed tests are robust to population admixture and are monotone increasing as a relative risk increases irrespective of mode of inheritance. We also illustrated the proposed tests with data adopted from Yonsei Cardiovascular Genome Center.

• Korean Journal of Materials Research
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• v.4 no.1
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• pp.9-23
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• 1994

Epitaxial $CoSi_2$ layer has been grown on NaCl(100) substrate at low deposition temperature($200^{\circ}C$) by multitarget bias cosputter deposition(MBCD). The phase sequence and crystallinity of deposited silicide as a function of deposition temperature and substrate bias voltage were studied by X-ray diffraction(XRD) and transmission electron microscopy(TEM) analysis. Crystalline Si was grown at $200^{\circ}C$ by metal induced crystallization(M1C) and self bias effect. In addition to, the MIC was analyzed both theoretically and experimentally. The observed phase sequence was $Co_2Si \to CoSi \to Cosi_2$ and was in good agreement with that predicted by effective heat of formation rule. The phase sequence, the CoSi(l11) preferred orientation, and the crystallinity had stronger dependence on the substrate bias voltage than the deposition temperature due to the collisional cascade mixing, the in-situ cleaning, and the increase in the number of nucleation sites by ion bombardment of growing surface. Grain growth induced by ion bombardment was observed with increasing substrate bias voltage at $200^{\circ}C$ and was interpreted with ion bombardment dissociation model. The parameters of $E_{Ar}\;and \alpha(V_s)$ were chosen to properly quantify the ion bombardment effect on the variation in crystallinty at $200^{\circ}C$ with increasing substrate bias voltage using Langmuir probe.

• Korean Journal of Microbiology
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• v.47 no.1
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• pp.14-21
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• 2011

Riboflavin synthase catalyzes the formation of one molecule of each riboflavin and 5-amino-6-ribitylamino-2,4-pyrimidinedione by the transfer of a 4-carbon moiety between two molecules of the substrates, 6,7-dimetyl-8-ribityllumazine. The most remarkable feature is the sequence similarity between the N-terminal half (1-97) and the C-terminal half domain (99-213). To investigate the structure and fluorescent characteristics of the N-terminal half of riboflavin synthase (N-RS) in Escherichia coli, more than 10 mutant genes coding for the mutated N-terminal domain of riboflavin synthase were generated by polymerase chain reaction. The genes coding for the proteins were inserted into pQE vector designed for easy purification of protein by 6X-His tagging system, expressed, and the proteins were purified. Almost all mutated N-terminal domain of riboflavin synthases bind to 6,7-dimethyl-8-ribityllumazine and riboflavin as fluorescent ligands. However, N-RS C47D and N-RS ET66,67DQ mutant proteins show colorless, indicating that fluorescent ligands were dissociated during purification. In addition, most mutated proteins show low fluorescent intensity comparing to N-RS wild type, whereas N-RS C48S posses stronger fluorescent intensity than that of wild type protein. Based on this result, N-RS C48S can be used as the tool for high throughput screening system for searching for the compound with inhibitory effect for the riboflavin synthase.

• The Korean Journal of Mycology
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• v.29 no.1
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• pp.34-40
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• 2001

RAPD test and the observation of morphological, cultural characteristics of fourteen selected entomogenous fungi were conducted to investigate the analysis of their internal relationships. Paecilomyces tenuipes showed snow flower form attached to numerous white conidiophores, produced globular and semi-egg types on the club types of phialides. Cordyceps militaris formed globosely conidiophores, dark yellow fruiting body on pupa. The phialide as on Acremonium-type in global conidiophores. Beauveria bassiana covered with conidia was not formed fruiting body and adhered conidia on conidiophore of zigzag type. The PDA and SDAY medium were confirmed as an optimum growth of them. P. tenuipes showed to velvet and plane types in several media whereas C. militaris was belong to centrally raised and floccose in the morphological type. In contrast, B. bassiana covered with conidia on velvet shape. The size of amplified products were analyzed by RAPD using URP primer and were from 100 bp to 2.0 kb with $10{\sim}14$ geuomic DNA. Total similarities of two groups were by dendrogram of UPGMA analysis. The homology of P. tenuipes groups was 94.8 to 100%. It also showed 70.1 to 96.6% in C. militaris group and B. bassiana was higher similarity than any other. The internal change of C. militaris, produced telemorph fruiting body, was higher seperated in species than P. tenuipes and B. bassiana in the RAPD.

• Restorative Dentistry and Endodontics
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• v.24 no.1
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• pp.13-25
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• 1999

P. endodontalis which was known to be associated with the infected root canals and periapical lesions is very difficult to detect by culture methods or traditional methods. Detection of bacteria using polymerase chain reaction(PCR) for 16S ribosomal DNA(rDNA) is fast, simple, and accurate with relatively small amount of target cells. 16S rDNA consist of conserved regions those are same to all species, and variable regions which represent species specificity. The 16S rDNA sequences of P. endodontalis and P. gingivalis were aligned and two highly variable regions were selected as a pair of species specific oligonucleotide primers for P. endodontalis. And then the pair of primers for PCR amplification was synthesized to identify P. endodontalis. The sequences of the species specific primers for the 16S rDNA of P. endodontalis were as follows ; sense primer[endo1]: 5'-CTATATTCTTCTTTCTCCGCATGGAGGAGG-3' antisense primer[endo2]: 5'-GCATACCTTCGGTCTCCTCTAGCATAT-3' In this study, for the identification of P. endodontalis without culture from the mixed clinical samples, PCR was done with species specific primers for the 16S rDNA sequences of P. endodontalis. The results were as follows : 1. The species specificity of the primers for the 16S rDNA of P. endodntalis was determined by the PCR methods. About 490bp amplicon which was specific only for P. endodntalis was produced with P. endodontalis. No amplicon was produced by PCR with other strains similar to P. endodontalis. 2. The synthesized species specific primers reacted with conventionally identified P. endodontalis which we have in conservative dentistry laboratory. 3. The identification of P. endodontalis using PCR technique with samples collected from infected root canals or periapical lesions was more sensitive than that of culture methods. 4. Seven samples revealed including P. endodontalis by PCR technique. Five of them were related with pains, two of them with sinus tract, three of them with foul odor, and three of them with purulent drainage. P. endodontalis was shown to have great relation with pains.

• Restorative Dentistry and Endodontics
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• v.24 no.1
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• pp.40-48
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• 1999

This study was designed to examine the specificities of the designed primers for F. nucleatum and F. necrophorum and to compare the PCR results using clinical samples with those of the anaerobic culture method. F. nucleatum and F. necrophorum spp. are frequently isolated in infected root canals, and they are related to periapical diseases. F. nucleatum(VPI 10197) and F. necrophorum(ATCC 25286) were used as references for PCR reaction, and thirty five teeth with one canal and periapical lesion were used. The samples were cultured anaerobically and identified using Rapid ID 32A(BioMerieux Vitek, Inc., France) as biochemical battery. In the GenBank database, species-specific PCR primers(nuc1/nuc2 primers for F. nucleatum and nec1/nec2 primers for F. necrophorum) were designed from the 16S ribosomal DNA(rDNA) sequences of F. nucleatum(accession number M58683) and F. necrophorum(accession number AF044948). PCR procedures of F. nucleatum(VPI 10197) and F. necrophorum (ATCC 25286) were simulated on a computer software. Amplify(v.1.2${\beta}$ for Macintosh). 820 bps and 817 bps of nucleotides were expected, respectively. Using extracted DNAs with QiaAmp tissue kit(Qiagen co., Germany), PCR was done. The results were as follows : 1. The nuc1/nuc2 primers produced an amplicon of 820 bps and the nec1/nec2 primers produced an amplicon of 817 bps. 2. The nuc1/nuc2 primers and the nec1/nec2 primers were specific and did not react with species other than the designated ones(i.e. nuc1/nuc2 primers did not produce amplicons for F. necrophorum, and vice versa.). And the PCR products of Porphyromonas endodontalis(ATCC 35406), Porphyromonas gingivalis(ATCC 33277), Prevotella intermedia(ATCC 25611), and Prevotella nigrescens(ATCC 33563), frequently isolated in infected root canals and periapical lesions, were not amplified by the primers specific for Fusobacterium nucleatum and Fusobacterium necrophorum. 3. This method utilizing PCR could detect F. nucleatum and F. necrophorum in clinical samples, while anaerobic culture method could detect neither.