• Clinical and Experimental Reproductive Medicine
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• v.47 no.1
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• pp.42-53
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• 2020

Objective: Recently, microRNA (miRNA) has been identified both as a powerful regulator involved in various biological processes through the regulation of numerous genes and as an effective biomarker for the prediction and diagnosis of various disease states. The objective of this study was to identify and validate miRNAs and their target genes involved in inflammation in placental tissue. Methods: Microarrays were utilized to obtain miRNA and gene expression profiles from placentas with or without inflammation obtained from nine normal pregnant women and 10 preterm labor patients. Quantitative real-time polymerase chain reaction and Western blots were performed to validate the miRNAs and differentially-expressed genes in the placentas with inflammation. Correlations between miRNA and target gene expression were confirmed by luciferase assays in HTR-8/SVneo cells. Results: We identified and validated miRNAs and their target genes that were differentially expressed in placentas with inflammation. We also demonstrated that several miRNAs (miR-371a-5p, miR-3065-3p, miR-519b-3p, and miR-373-3p) directly targeted their target genes (LEF1, LOX, ITGB4, and CD44). However, some miRNAs and their direct target genes showed no correlation in tissue samples. Interestingly, miR-373-3p and miR-3065-3p were markedly regulated by lipopolysaccharide (LPS) treatment, although the expression of their direct targets CD44 and LOX was not altered by LPS treatment. Conclusion: These results provide candidate miRNAs and their target genes that could be used as placental biomarkers of inflammation. These candidates may be useful for further miRNA-based biomarker development.

• Journal of Microbiology and Biotechnology
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• v.23 no.1
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• pp.1-6
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• 2013

Tetanus is a specific infectious disease, which is often associated with catastrophic events such as earthquakes, traumas, and war wounds. The obligate anaerobe Clostridium tetani is the pathogen that causes tetanus. Once the infection of tetanus progresses to an advanced stage within the wounds of limbs, the rates of amputation and mortality increase manifold. Therefore, it is necessary to devise a rapid and sensitive point-of-care detection method for C. tetani so as to ensure an early diagnosis and clinical treatment of tetanus. In this study, we developed a detection method for C. tetani using loop-mediated isothermal amplification (LAMP) assay, wherein the C. tetani tetanus toxin gene was used as the target gene. The method was highly specific and sensitive, with a detection limit of 10 colony forming units (CFU)/ml, and allowed quantitative analysis. While detecting C. tetani in clinical samples, it was found that the LAMP results completely agreed with those of the traditional API 20A anaerobic bacteria identification test. As compared with the traditional API test and PCR assay, LAMP detection of C. tetani is simple and rapid, and the results can be identified through naked-eye observation. Therefore, it is an ideal and rapid point-of-care testing method for tetanus.

• The Plant Pathology Journal
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• v.31 no.4
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• pp.371-378
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• 2015

Surveys of yellowing viruses in plastic tunnels and in open field crops of melon (Cucumis melo cultivar catalupo), oriental melon (C. melo cultivar oriental melon), and cucumber (C. sativus) were carried out in two melon-growing areas in 2014, Korea. Severe yellowing symptoms on older leaves of melon and chlorotic spots on younger leaves of melon were observed in the plastic tunnels. The symptoms were widespread and included initial chlorotic lesions followed by yellowing of whole leaves and thickening of older leaves. RT-PCR analysis using total RNA extracted from diseased leaves did not show any synthesized products for four cucurbit-infecting viruses; Beet pseudo-yellows virus, Cucumber mosaic virus, Cucurbit yellows stunting disorder virus, and Melon necrotic spot virus. Virus identification using RT-PCR showed Cucurbit aphid-borne yellows Virus (CABYV) was largely distributed in melon, oriental melon and cucumber. This result was verified by aphid (Aphis gossypii) transmission of CABYV. The complete coat protein (CP) gene amplified from melon was cloned and sequenced. The CP gene nucleotide and the deduced amino acid sequence comparisons as well as phylogenetic tree analysis of CABYV CPs showed that the CABYV isolates were undivided into subgroups. Although the low incidence of CABYV in infections to cucurbit crops in this survey, CABYV may become an important treat for cucurbit crops in many different regions in Korea, suggesting that CABYV should be taken into account in disease control of cucurbit crops in Korea.

• The Plant Pathology Journal
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• v.26 no.4
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• pp.328-339
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• 2010

A tobamovirus, Ribgrass mosaic virus (RMV), was identified newly from chinese cabbage (Brassica campestris L. pekinensis) in Korea. Virus disease incidence of RMV on chinese cabbage was 37.9% in alpine area on August in 1993. RMV induced the symptoms of necrotic ring spots, necrotic streak on midrib and malformation. RMV, Ca1 and Ca3 isolate, could infect 35 species out of 45 plants including Chenopodium amaranticolor. Physical properties of RMV Ca1 isolate were very stable as 10.8 over for dilution end point, $95^{\circ}C$ for temperature inactivation point and 18 weeks for longevity in vitro. RMV had the soil transmission rate of 75.0% for the chinese cabbages, 'Chunhawang' and 'Seoul' cultivars. The purified virions of RMV had the typical ultraviolet absorption spectrum of maximum at 260 nm and minimum at 247 nm. RMV of Ca1 isolate was related serologically with antisera of Tobacco mosaic virus (TMV)-Cym, TMV-O and Pepper mottle virus, but not related with antiserum of Odontoglossum ring spot virus. coat protein gene of RMV-Ca1, sized 473 nucleotides, encoded 158 amino acid residues. Nucleotide identity of RMV-Ca1 CP gene was 96.4% with RMV-Shanghai (GenBank accession No. of AF185272) from China and 96.0% with RMV-Impatiens (GenBank accession No. of AM040974) from Germany. Identity of amino acids between RMV-Ca1 and the two RMV isolates was 96.8%. Specific three primers were selected for rapid and easy genetic detection of RMV using Virion Captured (VC)/RT-PCR method.

• Neonatal Medicine
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• v.28 no.3
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• pp.133-138
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• 2021

Osteopetrosis refers to a group of genetic skeletal disorders characterized by osteosclerosis and fragile bones. Osteopetrosis can be classified into autosomal dominant, autosomal recessive, or X-linked forms, which might differ in clinical characteristics and disease severity. Autosomal recessive osteopetrosis, also known as malignant osteopetrosis, has an earlier onset, more serious clinical symptoms, and is usually fatal. We encountered a 1-day-old girl who was born full-term via vaginal delivery, which was complicated by meconium-stained amniotic fluid, cephalo-pelvic disproportion, and nuchal cord. Routine neonatal care was provided, in addition to blood tests and chest radiography to screen for sepsis, as well as skull radiography to rule out head injuries. Initial blood tests revealed hypocalcemia, which persisted on follow-up tests the next day. Radiographic examinations revealed diffusely increased bone density and a "space alien" appearance of the skull. Based on radiographic and laboratory findings, the infantile form of osteopetrosis was suspected and genetic testing for identification of the responsible gene. Eventually, a heterozygous mutation of the T cell immune regulator 1, ATPase H+ transporting V0 subunit a3 (TCIRG1) gene (c.292C>T) was identified, making this the first reported case of neonatal-onset malignant osteopetrosis with TCIRG1 mutation in South Korea. Early-onset hypocalcemia is common and usually results from prematurity, fetal growth restriction, maternal diabetes, perinatal asphyxia, and physiologic hypoparathyroidism. However, if hypocalcemia persists, we recommend considering 'infantile of osteopetrosis' as a rare cause of neonatal hypocalcemia and performing radiographic examinations to establish the diagnosis.

• The Plant Pathology Journal
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• v.38 no.1
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• pp.1-11
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• 2022

Fusarium culmorum is one of the most important causal agents of root rot of wheat. In this study, 10 F. culmorum isolates were collected from farms located in five agro-ecological regions of Morocco. These were used to challenge 20 durum wheat genotypes via artificial inoculation of plant roots under controlled conditions. The isolate virulence was determined by three traits (roots browning index, stem browning index, and severity of root rot). An alpha-lattice design with three replicates was used, and the resulting ANOVA revealed a significant (P < 0.01) effect of isolate (I), genotype (G), and G × I interaction. A total of four response types were observed (R, MR, MS, and S) revealing that different genes in both the pathogen and the host were activated in 53% of interactions. Most genotypes were susceptible to eight or more isolates, while the Moroccan cultivar Marouan was reported resistant to three isolates and moderately resistant to three others. Similarly, the Australian breeding line SSD1479-117 was reported resistant to two isolates and moderately resistant to four others. The ICARDA elites Icaverve, Berghisyr, Berghisyr2, Amina, and Icaverve2 were identified as moderately resistant. Principal component analysis based on the genotypes responses defined two major clusters and two sub-clusters for the 10 F. culmorum isolates. Isolate Fc9 collected in Khemis Zemamra was the most virulent while isolate Fc3 collected in Haj-Kaddour was the least virulent. This work provides initial results for the discovery of differential reactions between the durum lines and isolates and the identification of novel sources of resistance.

• Journal of Genetic Medicine
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• v.19 no.2
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• pp.63-75
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• 2022

Purpose: Osteoporosis is a common calcium and metabolic skeletal disease which is characterized by decreased bone mass, microarchitectural deterioration of bone tissue and impaired bone strength, thereby leading to enhanced risk of bone fragility. In this study, we aimed to identify novel genes for susceptibility to osteoporosis and/or bone density. Materials and Methods: To identify differentially expressed genes (DEGs) between control and osteoporosis-induced cells, annealing control primer-based differential display reverse-transcription polymerase chain reaction (RT-PCR) was carried out in pre-osteoblast MC3T3-E1 cells. Expression levels of the identified DEGs were evaluated by quantitative RT-PCR. Association studies for the quantitative bone density analysis and osteoporosis case-control analysis of single nucleotide polymorphism (SNPs) were performed in Korean women (3,570 subjects) from the Korean Association REsource (KARE) study cohort. Results: Comparison analysis of expression levels of the identified DEGs by quantitative RT-PCR found seven genes, Anxa6, Col5a1, Col6a2, Eno1, Myof, Nfib, and Scara5, that showed significantly different expression between the dexamethason-treated and untreated MC3T3-E1 cells and between the ovariectomized osteoporosis-induced mice and sham mice. Association studies revealed that there was a significant association between the SNPs in the five genes, ANXA6, COL5A1, ENO1, MYOF, and SCARA5, and bone density and/or osteoporosis. Conclusion: Using a whole-genome comparative expression analysis, gene expression evaluation analysis, and association analysis, we found five genes that were significantly associated with bone density and/or osteoporosis. Notably, the association P-values of the SNPs in the ANXA6 and COL5A1 genes were below the Bonferroni-corrected significance level.

• 한국균학회소식:학술대회논문집
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• 2014.05a
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• pp.41-41
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• 2014

Pinewood nematode (PWN, Bursaphelenchus xylophilus) is a serious pathogenic worm that quickly dry pine trees to death. Recently, PWN has been devastating huge amounts of conifer trees in Korea. As a first step to explore the association and ecological roles of fungi in PWN life cycle in Korea, in this study we first isolated and indentified fungi from PWN-infested Korean pine and Japanese black pine wood sampled in Jinju, Sacheon, Pocheon, Chuncheon, Gwangju, and Hoengseong in Korea. A total of 144 fungal isolates were obtained from Japanese black pine wood and 264 fungal isolates from Korean pine wood. Their morphology and nucleotide sequences of the ITS rDNA and ♌-tubulin gene were examined for species identification. Ophiostoma ips, Botrytis anthophila, Penicillium sp., Hypocrea lixii, Trichoderma atroviride, O. galeiforme, Fusarium proliferatum were identified from Japanese black pine wood. Leptographium koreanum, L. pini-densiflorae, Ophiostoma ips, Penicillium raistrick, Trichoderma sp. were isolated from Korean pine wood. O. ips and L. koreanum were the major species on the two different PWN-infected pine tree. The cultivation of PWN on fungal mat of the identified species did some enhance PWN reproduction. The ambrosia beetle, Platypus koryoensis, is a serious pest of oak trees in Korea. In this study we investigated filamentous fungi present in the body of the beetle. Fourteen genera of filamentous fungi belonging to Ascomycota and Basidiomycota were isolated. All the obtained genera were isolated in the mitosporic state. The identified fungi were classified in 11 distinct orders including the Ascomycota (Eurotiales, Hypocreales, Microascales, Ophiostomatales, Pleosporales, and Sordiales) and Basidiomycota (Agaricales, Corticiales, Polyporales, and Russulales Xylariales). Within Ascomycota, 13 species were found. Meanwhile five species were found within Basidiomycota. The results showed the presence of diverse fungi in P. koryoensis. Among the isolated fungi, some were able to produce wood degrading enzymes. Further fungal isolation was performed with P. koryoensis infested Quercus mongolica trees sampled at Kumdan mountain in Hanam-Si, Gyeonggi province from June of 2009 to June of 2010. Penicillin spp. and Trichoderma spp. were the major species of mold fungi group. Pichia guilliermondii was the major species of mold yeast group. Raffaelea quercus-mongolicae was also isolated, but its isolation frequency was not high. Other species identified were Ambrosiella xylebori, Fusarium solani, Cryphonectria nitschke, Chaetomium globosum, and Gliocladium viride, Candida kashinagacola, C. maritima, C. vanderkliftii, Saccharomycopsis crataegensis.

• The Plant Pathology Journal
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• v.24 no.2
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• pp.207-210
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• 2008

In March of 2003, tumors (galls) were observed on ginseng seedling roots in ginseng seedbeds at Yeoju, Gyeonggi province, Korea. Symptoms were spherical or galls with about 0.5-1.0cm in diameter formed on the upper through middle parts of the primary roots. Bacterial isolates obtained from the root galls were Gram-negative, rod-shaped with peritrichous flagella, aerobic, not forming yellow or orange colonies on nutrient glucose agar, yeast extract-dextrose $CaCO_3$ agar and nutrient-broth yeast extract agar, non-fluorescent on King's B agar, and non-spore forming, which were identical to characteristics of the genus Agrobacterium. They were identified as Agrobacterium tumefaciens with 0.732-0.993 similarities in 100% probability by the Biolog analyses. The 16S rRNA gene partial sequences of the six isolates tested (Genbank Accession EF486308-EF486313) were 100% homologous to those of other A. tumefaciens strains (GenBank accession AF501343, AY701900, AY701898, AY701899). The above results confirmed that this bacterium is A. tumefaciens. Pathogenicity of the bacteria was proved by the inoculation test on carrot root discs and tomato seedlings. This is the first description of A. tumefaciens causing root gall in ginseng seedling. The disease occurred locally and sparsely, but considering its appearances in seedbeds suggests that the ginseng root gall may become a threat to ginseng in Korea.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.12
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• pp.1830-1836
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• 2003

Studies using natural resistance-associated macrophage protein (NRAMP) identification indicated that cattle could be selected for immunity. Several studies performed on intracellular organisms such as Mycobacterium, Salmonella, Brucella and Leishmania in human and mouse revealed that resistance against these bacteria was dependent on high activity of NRAMP1 in macrophages. However, hardly any researches have been done on Staphylococcus aureus in bovine mastitis, which is an intracellular organism and the main cause of bovine mastitis. The objectives of this study were to establish reverse transcriptase polymerase chain reaction (RT-PCR) methods, through which NRAMP1 mRNA expression could be compared and analyzed between mastitis-resistant and -susceptible cows. NRAMP1 gene and its expression were investigated using 20 cows (Holstein Friesian) in Korea. Cows were evenly split into two groups, with and without histories of clinical mastitis. Equivalent numbers of cows were randomly selected from each group. Monocytes were isolated from the bovine peripheral blood of each selected cows and activated with lipopolysaccharide (LPS). mRNA was separated from the monocytes and cDNA of NRAMP1 was synthesized and amplified using RT-PCR with amplification of $\beta$-actin as a control. The difference in NRAMP1 expressions of mastitis-resistant (n=10) and -susceptible (n=10) Holstein cows was analyzed. Results demonstrate that resistant cows produced more NRAMP1 mRNA than the susceptible ones, and ratios of NRAMP1:$\beta$-actin expression were higher in resistant cows with or without LPS activation. Therefore, this study could be applied to select bovine mastitis resistant cows before infection based on the expression of NRAMP1.