In the post-genome period, the technique for identifying gene expression has been progressed to high throughput screening. In the field of molecular nutrition, the use of screening techniques to clarify molecular function of specific nutrients would be very advantageous. In this study, we have evaluated Zn-regulated gene expression in Zn-deficient, homocystein-treated EA.hy926 cells, using cDNA microarray, which can be used to screen the expression of many genes simultaneously. The information obtained can be used for preliminary assessment of molecular and signaling events modulated by Zn under pro-atherogenic conditions. EA.hy926 cells derived from human umbilical vein endothelial cells were cultured in Zn-adequate (control, 15 $\mu$M Zn) or Zn-deficient (experimental, 0 $\mu$M Zn) Dulbecco's MEM media under high homocysteine level (100 $\mu$M) for 3 days of post-confluency. Cells were harvested and RNA was extracted. Total RNA was reverse-transcribed and the synthesized cDNA was labeled with Cy3 or Cy5. Fluorescent labeled cDNA probe was applied to microarray slides for hybridization, and the slide was then scanned using a fluorescence scanner. The expression of seven genes was found to be significantly decreased, and one significantly increased, in response to treatment of EA.hy926 cells with Zn-deficient medium, compared with Zn-supplemented medium. The upregulated genes were oncogenes and tumor suppressor genes, cell cycle-related genes and transporter genes. The down-regulated gene was RelB, a component of the NF-kappaB complex of transcription factors. The results of this study imply the effectiveness of cDNA microarray for expression profiling of a singly nutrient deficiency, namely Zn. Furthur study, using tailored-cDNA array and vascular endothelial cell lines, would be beneficial to clarify the molecular function of Zn in atherosclerosis, more in detail.
RAPD analysis showed that all the OTUs of 'Sandolbae' were the same species because amplified band patterns of all samples generated by each of 5 random primers were identical. Even though there were different environmental conditions, all the "Chuiangne" trees from three different places were the same species, and also all the "Cheongshilli" trees were the same species too. No genetic variations were detected between native Korean pears grown in the habitats and in the research field. Because 212 polymorphic bands were generated by 9 primers selected through primer screening, they were possible to analyze genetic relationship among naturally growing three native Korean pears and nine cultivars of Pyrus pyrifolia and P. communis. Based on the RAPD analysis, three main groups were formed. The first group represented the Six P. pyrifoia cultivars, the second group was the three native Korean pears, and the last group was the three P. communis cultivars. Genetic distance between 'Wonwhang' and 'Chojuro' was closer than other cultivars in group 1 since dissimilarity index value between these two cultivars was 50.82. However, genetic distance between 'Niitaka' and 'Chojuro' was the most distant compared to the others in group 1. In group 2, 'Sandlobae' was genetically closer to 'Chuiangne' than 'Cheongshilli' because dissimilarity index value between 'Sandlobae' and 'Chuiangne' was smaller, 50.82, than the value between 'Sandlobae' and 'Cheongshilli', 63.636. In group 3, 'Old Home' was genetically closer to 'Bartlett' than 'Kaiser Alexander(or Bosc)'. Group 3 composed of P. communis cultivars was genetically further than other two groups, P. pyrifolia cultivars and native Korean pears.
Although financial information is a great influence upon determining of the group which use them, detection of management fraud and earning manipulation is a difficult task using normal audit procedures and corporate credit evaluation processes, due to the shortage of knowledge concerning the characteristics of management fraud, and the limitation of time and cost. These limitations suggest the need of systemic process for !he effective risk of earning manipulation for credit evaluators, external auditors, financial analysts, and regulators. Moot researches on management fraud have examined how various characteristics of the company's management features affect the occurrence of corporate fraud. This study examines financial characteristics of companies engaged in fraudulent financial reporting and suggests a model and system for detecting GAAP violations to improve reliability of accounting information and transparency of their management. Since the detection of management fraud has limited proven theory, this study used the detecting method of outlier(upper, and lower bound) financial ratio, as a real-field application. The strength of outlier detecting method is its use of easiness and understandability. In the suggested model, 14 variables of the 7 useful variable categories among the 76 financial ratio variables are examined through the distribution analysis as possible indicators of fraudulent financial statements accounts. The developed model from these variables show a 80.82% of hit ratio for the holdout sample. This model was developed as a financial outlier detecting system for a financial institution. External auditors, financial analysts, regulators, and other users of financial statements might use this model to pre-screen potential earnings manipulators in the credit evaluation system. Especially, this model will be helpful for the loan evaluators of financial institutes to decide more objective and effective credit ratings and to improve the quality of financial statements.
In post-genome period, the technique for identifying gene expression has been changed to high throughput screening. In the field of molecular nutrition, the need for this technique to clarify molecular function of the specific nutrient is essential. In this study, we have tested the zinc-regulated gene expression in zinc-deficient U937 cells, using cDNA microarray which is the cutting-edge technique to screen large numbers of gene expression simultaneously. The study result can be used for the preliminary gene screening data for clarifying, using monocyte U937 cell line, molecular Zn aspect in atherosclerosis. U937 cells were cultured in Zn-adequate (control, 12 $\mu$M Zn) or Zn-deficient (experimental, 0 $\mu$M Zn) ESMI media during 2 days, respectively. Cells were harvested and RNA was extracted. Total RNA was reverse-transcriptinized and synthesized cDNA probe labeled with Cy-3. fluorescent labeled cDNA probe was applied to microarray slide for hybridization slide, and after then, the slide was scanned using fluorescence scanner. ‘Highly expressed genes’ in Zn-deficient U937 cells, comparing to Zn-adequate group, are mainly about the genes for motility protein, immune system protein, oncogene and tumor suppressor and ‘Less highly expressed genes’ are about the genes for transcription, apoptosis associated protein, cell cycle, and several basic transcription factors. The results of this preliminary study imply the effectiveness of cDNA microarray for expression profiling of a singly nutrient deficiency, specially Zn. Furthur study, using tailored-cDNA array and capillary endothelial cell lines, would be beneficial to clarify molecular Zn function, more in detail.
This experiment was conducted to find out an effective propagation method for reed(Phragmites communis Trin.), ensuring a continuous herbicide screening for reed control. Reed propagation methods were compared under a greenhouse condition using tour different materials; seeds, rhizomes, depressed stolons of P. japonica Steud., and stem cuttings. Although reed seeds were easy to harvest and store, their germination rate(${\leq}$5%) was very low and seedling growth from the seeds was slow. Rhizomes were difficult to harvest and their harvest time was limited from November to March. Furthermore, reed propagation using rhizomes had problems of a relatively low germination rate(46%), no uniformity in size and shape, individual differences at the early stage of growth, and difficulties in material storage. Rate of reed growth from rhizomes was higher in commercial soil mix(Boo Nong soil) than in sand or in sand+upland soil(1:1). Depressed stolons of P. japonica had a moderate germination rate(65%) and were relatively easy to harvest. However, their harvest time was limited only from August to September. Propagation method using stem cuttings had several advantages over the above methods using other materials. Reed plants could uniformly be propagated from the stem cuttings with a relatively high germination rate(75%). Stem cuttings of central nodes showed a higher germination rate compared to those of upper or lower nodes. Stem cuttings from the field should be used immediately after harvest, since their germination rate decreased rapidly when they were stored under a wet- or a dry-refrigerated condition. Furthermore, the germination of stem cuttings tended to decrease when they were collected from the field after August. This indicates that there is a limitation of harvest time for stem cuttings. However, a year-round propagation of reed using stem cuttings is possible if parent plants are grown in a greenhouse, and thus herbicide screening for reed control could continuously be performed.
Song, Jang-Hoon;Md, Abdul Alim;Choi, Eu-Ddeum;Choi, Duck-Soo;Seo, Ho-Jin
Korean Journal of Organic Agriculture
/
v.26
no.2
/
pp.259-268
/
2018
The study investigated the efficacy of four different essential oils on the repellent responses of large black chafer (Holotrichia parallela) Motschulsky (Coleoptera: Scarabaeidae) in organic pear (Pyrus pyrifolia) orchards. Cinnamon, pine, peppermint, and eucalyptus oils were used, and the behavioral responses and repellent effects against H. parallela were investigated under laboratory and field conditions. Adult beetle responses to different oils were examined using a Y-tube olfactometer in the laboratory and four absorbent blocks with each oil in the field. The repellent responses rates of H. parallela were 100% for cinnamon oil; however, only 67% of adult beetles avoided peppermint and eucalyptus oil in the Y-tube olfactometer bioassay. In the field tests, the least damage to leaves was observed on trees treated with cinnamon oil, whereas the most damage was observed in the control (non-treated) trees and those treated with peppermint oil, followed by eucalyptus and pine oil. Therefore, cinnamon oil can be used as a repellent to avoid damage form large black chafers in organic pear orchards.
Bt-cotton germplasm, consisting of 75 genotypes was evaluated against cotton leaf curl disease (CLCuD) under high inoculum pressure in the field and using graft inoculation in glasshouse by visual symptom scoring assessments. None of the tested genotype was found disease free under both evaluation tests. Under field conditions in 2011, 3 genotypes were found resistant, 4 moderately resistant, 3 tolerant, 2 moderately susceptible and one susceptible; in 2012, 3 genotypes were tolerant, 7 moderately susceptible, 5 susceptible and 38 highly susceptible; in 2013, one was moderately susceptible and 51 were highly susceptible with varying degree of percent disease index (PDI) and severity index (SI). However, through graft evaluation in glasshouse, none of the graft inoculated plant was symptomless. All tested genotypes showed disease symptoms with SI values ranging between 5.0 and 6.0, and latent period between 12 and 14 days. Of the 75 genotypes evaluated using graft inoculation, 11 were found susceptible with SI values of 5.0 to 5.4 while remaining 64 were highly susceptible with SI values of 5.5 to 6.0. Inoculated plants of all tested genotypes exhibited severe disease symptoms within 10 days after the appearance of initial symptoms. No reduction in SI value was observed until the end of the experiment i.e., 90 days after grafting. Information generated under the present study clearly demonstrates that no sources of resistance to CLCuD are available among the tested Bt-cotton genotypes. So, a breeding programme is needed to introgress the CLCuD-resistance from other resistant sources to agronomically suitable Bt-cotton genotypes.
Choe, Hyeokmin;Nho, Hyun Woo;Park, Jonghoon;Kim, Jin Bae;Yoon, Tae Hyun
Bulletin of the Korean Chemical Society
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v.35
no.2
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pp.365-370
/
2014
For real-time monitoring of colloidal nanoparticles (NPs) in aqueous media, a light sheet type dark-field microscopy system combined with a microfluidic concentration gradient generator (${\mu}FCGG$-LSDFM) was developed. Various concentrations of colloidal Au NPs were simultaneously generated with the iFCGG and characterized with the LSDFM setup. The number concentrations and hydrodynamic size distributions were measured via particle counting and tracking analysis (PCA and PTA, respectively) approaches. For the 30 nm Au NPs used in this study, the lower detection limit of the LSDFM setup was 3.6 ng/mL, which is about 400 times better than that of optical density measurements under the same ${\mu}FCGG$ system. Additionally, the hydrodynamic diameter distribution of Au NPs was estimated as $39.7{\pm}12.2nm$ with the PTA approach, which agrees well with DLS measurement as well as the manufacturer's specification. We propose this ${\mu}FCGG$-LSDFM setup with features of automatic generation of NP concentration gradient and real-time monitoring of their physicochemical characteristics (e.g., number concentration, and hydrodynamic size distribution) as an important component of future high-throughput screening or high-content analysis platforms of nanotoxicity.
Kim, Hun;Jo, Eun Ju;Choi, Yong Ho;Jang, Kyoung Soo;Choi, Gyung Ja
The Plant Pathology Journal
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v.32
no.5
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pp.423-430
/
2016
Clubroot disease caused by Plasmodiophora brassicae is one of the most serious diseases in Brassica crops worldwide. In this study, the pathotypes of 12 Korean P. brassicae field isolates were determined using various Chinese cabbage including 22 commercial cultivars from Korea, China, and Japan, and 15 inbred lines. All P. brassicae isolates exhibited the typical clubroot disease on non-clubroot resistant cultivar, indicating that the isolates were highly pathogenic. According to the reactions on the Williams' hosts, the 12 field isolates were initially classified into five races. However, when these isolates were inoculated onto clubroot-resistant (CR) cultivars of Chinese cabbage, several isolates led to different disease responses even though the isolates have been assigned to the same race by the Williams' host responses. Based on the pathogenicity results, the 12 field isolates were reclassified into four different groups: pathotype 1 (GN1, GN2, GS, JS, and HS), 2 (DJ and KS), 3 (HN1, PC, and YC), and 4 (HN2 and SS). In addition, the CR cultivars from Korea, China, and Japan exhibited distinguishable disease responses to the P. brassicae isolates, suggesting that the 22 cultivars used in this study, including the non-CR cultivars, are classified into four different host groups based on their disease resistance. Combining these findings, the four differential hosts of Chinese cabbage and four pathotype groups of P. brassicae might provide an efficient screening system for resistant cultivars and a new foundation of breeding strategies for CR Chinese cabbage.
Tirumalareddy Danda;Jong-Won Park;Kimberly L. Timmons;Mamoudou Setamou;Eliezer S. Louzada;Madhurababu Kunta
The Plant Pathology Journal
/
v.39
no.4
/
pp.309-318
/
2023
Huanglongbing (HLB) is one of the most destructive diseases in citrus, which imperils the sustainability of citriculture worldwide. The presumed causal agent of HLB, 'Candidatus Liberibacter asiaticus' (CLas) is a non-culturable phloem-limited α-proteobacterium transmitted by Asian citrus psyllids (ACP, Diaphorina citri Kuwayama). A widely adopted method for HLB diagnosis is based on quantitative real-time polymerase chain reaction (qPCR). Although HLB diagnostic qPCR provides high sensitivity and good reproducibility, it is limited by time-consuming DNA preparation from plant tissue or ACP and the requirement of proper lab instruments including a thermal cycler to conduct qPCR. In an attempt to develop a quick assay that can be deployed in the field for CLas detection, we developed a real-time loop-mediated isothermal amplification (rt-LAMP) assay by targeting the CLas five copy nrdB gene. The rt-LAMP assay using various plant sample types and psyllids successfully detected the nrdB target as low as ~2.6 Log10 copies. Although the rt-LAMP assay was less sensitive than laboratory-based qPCR (detection limit ~10 copies), the data obtained with citrus leaf and bark and ACP showed that the rt-LAMP assay has >96% CLas detection rate, compared to that of laboratory-based qPCR. However, the CLas detection rate in fibrous roots was significantly decreased compared to qPCR due to low CLas titer in some root DNA sample. We also demonstrated that the rt-LAMP assay can be used with a crude leaf DNA extract which is fully deployable in the field for quick and reliable HLB screening.
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