• Journal of Life Science
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• v.18 no.8
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• pp.1023-1030
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• 2008

NtMEK2, which is the tobacco MAPK kinase that is upstream of SIPK and WIPK, was identified using the dexamethasone (DEX)-inducible gain-of-function transgenic system. Expression of $NtNEK2^{DD}$, a constitutively active mutant of NtNEK2, leads to HR-like cell death, which indicates that the NtMEK2-SIPK/WIPK cascade controls defense responses in tobacco. However, little is known about the downstream target substrates or defense-related genes that are regulated by the NtMEK2-SIPK/ WIPK cascade. In this study, ACP-based differential display RT-PCR was used to isolate the downstream effectors mediated by the NtMEK2-SIPK/WIPK cascade in $NtNEK2^{DD}$ transgenic plants. The results identified 6 novel differentially expressed genes (DEGs). These included pathogen induced protein 2-4 (pI2-4), monoterpene synthase 2 (MTS2), seven in absentia protein (SINA), cell death marker protein 1 (CDM1), hydroxyproline-rich glycoprotein (HRGP) and unknown genes (DEG45). The induction of these genes was confirmed by RT-PCR of samples obtained from $NtNEK2^{DD}$ plants. Additionally, when compared with other isolated DEGs, the pI2-4, CDM1 and HRGP genes were significantly up-regulated in response to treatment with salicylic acid and tobacco mosaic virus. Taken together, these results suggest that three novel DEGs were regulated by the NtMEK2-SIPK/WIPK cascade involved in disease resistance in tobacco.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.47 no.6
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• pp.395-401
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• 2002

The combination of early heading time, maturing time and short grain-filling period is very important to develop early varieties in winter barley. The 4 parental half diallel crosses (parents, $F_1$s, $F_2$s) were cultivated at the field. The heading date was from April 3 to 26, maturing date from May 15 to 27 and grain-filling period from 31 days to 42 days, showing that the varietal differences about the 3 traits were remarkable. According to half diallel cross analyses, Dongbori 1 for heading time (late heading) was dominant, but Oweolbori (early heading) was recessive, showing partial dominance with high additive component of genetic variance. Dongbori 1 for maturing time was dominant, but Oweolbori was recessive, showing partial dominance with high additive variance. Reno for grain-filling period (short grain-filling period) was dominant, but Oweolbori (long grain-filling period) was recessive with additive, and partial dominance. There were highly significant mean squares for both GCA and SCA effects on the heading and maturing times, and GCA/SCA ratios for all traits were high, showing the additive gene effects more important. Sacheon 6 and Oweolbori had greater GCA effects for early heading and maturing times, and Dongbori 1 and Reno had greater GCA effects for late times. GCA effects were highly significant in $F_1$ and $F_2$ generations, showing high GCA/SCA ratios (7.02). The heading and maturing times in field were positively correlated with antifreeze proteins concentrations, accumulation, resistance to photoinhibition and winter survival, respectively) but the grain-filling period did negatively correlated with the trails.

• Journal of Genetic Medicine
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• v.4 no.2
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• pp.160-166
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• 2007

Purpose : The ${\beta}$-adrenoceptors are pharmacologically classified into ${\beta}_1$-, ${\beta}_2$- and ${\beta}_3$-adrenoceptor. The gene of each subtype has polymorphisms related to their function (${\beta}_1$-adrenoceptor: Ser49Gly, ${\beta}_2$-adrenoceptor: Gln27Glu, ${\beta}_3$-adrenoceptor: Trp64Arg). The objectives of this study were to analyse the allelic and genotypic distribution of the representative polymorphism of ${\beta}$-adrenoceptors in preeclampsia and to investigate whether combined genotype of ${\beta}$-adrenoceptors may be associated with preeclampsia. Methods : Blood samples were collected from a Korean population (159 preeclamptic pregnancies and 168 normotensive pregnancies). The ${\beta}_1$-, ${\beta}_2$- and ${\beta}_3$-adrenoceptor genotypes was determined using polymerase chain reaction-restriction fragment length polymorphism. Results : There were no differences in allelic and genotypic distribution of ${\beta}_1$- and ${\beta}_2$-adrenoceptor polymorphisms between the two groups. However, the Arg allele of ${\beta}_3$-adrenoceptor polymorphism were more frequent in preecalmpsia than in controls (P<0.05, OR=1.57, 95% CI=1.01-2.46). Moreover, prevalence of genotype carrying heterozygote of ${\beta}_3$-adrenoceptor polymorphism was increased in preeclampsia compared with controls (P<0.05, OR 1.76, 95% CI 1.06-2.92). When combination of the three polymorphisms were evaluated, pregnancies with the particular combined genotype that is consisted of heterozygote of ${\beta}_1$-, ${\beta}_3$-adrenoceptor and wild homozygote of ${\beta}_2$-adrenoceptor (Ser/Gly, Gln/Gln, Trp/Arg), showed a significant increase in the risk of preeclampsia (P<0.05, OR=3.01, 95% CI 1.12-8.08). Conclusion : A particular combined genotype (Ser/Gly, Gln/Gln, Trp/Arg) of - adrenoceptors was associated with the risk of preeclampsia.

• Journal of Plant Biotechnology
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• v.39 no.1
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• pp.33-48
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• 2012

As a scientific curiosity to understand the structure and the function of crops and experimental efforts to apply it to plant breeding, genetic maps have been constructed in various crops. Especially, in the case of Brassica crop, genetic mapping has been accelerated since genetic information of model plant $Arabidopsis$ was available. As a result, the whole $B.$ $rapa$ genome (A genome) sequencing has recently been done. The genome sequences offer opportunities to develop molecular markers for genetic analysis in $Brassica$ crops. RFLP markers are widely used as the basis for genetic map construction, but detection system is inefficiency. The technical efficiency and analysis speed of the PCR-based markers become more preferable for many form of $Brassica$ genome study. The massive sequence informative markers such as SSR, SNP and InDels are also available to increase the density of markers for high-resolution genetic analysis. The high density maps are invaluable resources for QTLs analysis, marker assisted selection (MAS), map-based cloning and comparative analysis within $Brassica$ as well as related crop species. Additionally, the advents of new technology, next-generation technique, have served as a momentum for molecular breeding. Here we summarize genetic and genomic resources and suggest their applications for the molecular breeding in $Brassica$ crop.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.5
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• pp.761-767
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• 2007

This study was conducted to determine the effects of exogenous zinc-metallothionein (Zn-MT) on anti-oxidative function and pork quality. After feeding a corn-soybean meal-based diet for two weeks, 48 pigs ($Duroc{\times}Landrace{\times}Chinese\;Black Pig$) were assigned randomly to four groups. Pigs in Group 1 were maintained under non-stress conditions, whereas pigs in Groups 2, 3 and 4 were aggressively handled for 25 min to produce stress. Pigs in Groups 1, 2, 3, and 4 received intramuscular administration of saline (control group; CON), 0 (negative control group; NCON), 0.8 (low dose group; LOW), and 1.6 (high dose group; HIGH) mg rabbit liver Zn-MT per kg body weight, respectively. Pigs were slaughtered at 3 and 6 h post-injection. Zn-MT treatment increased (p<0.05) the activities of superoxide dismutase (SOD) and glutathione-peroxidase (GSH-PX) while decreasing the concentration of malondialdehyde (MDA) in liver. These responses were greater (p<0.05) at 6 h than at 3 h post Zn-MT injection. Zn-MT treatment increased (p<0.05) hepatic SOD mRNA levels in a time and dose-dependent manner and decreased (p<0.05) serum glutamate-pyruvate transaminase and lactate dehydrogenase activities (indicators of tissue integrity). Zn-MT administration decreased (p<0.05) lactate concentration and increased (p<0.05) pH and water-holding capacity in the longissimus thorasis meat. Collectively, our results indicate that intramuscular administration of Zn-MT to pre-slaughter stressed pigs improved tissue anti-oxidative ability and meat quality.

• Journal of Korean Biological Nursing Science
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• v.2 no.1
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• pp.1-19
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• 2000

The purpose of this study was to define the content of the requisite knowledge of pathophysiology, clinical microbiology, and mechanisms and effects of drugs needed for clinical knowledge for nursing practice. Contents of knowlege on pathological physiology, clinical microbiology, and mechanisms and effects of drugs were constructed from syllabus of basic nursing subjects in 4 colleges of nursing, and textbooks. The degree of need of 72 items was measured with a 4 point scale. The subjects of this study were college-graduated 136 nurses from seven university hospital in Seoul and three in Chonnam Province, Kyungbook Province, and Inchon. They have been working at internal medicine ward, surgical ward, intensive care unit, obstetrics and gynecology ward, pediatrics ward, opthalmology ward, ear, nose, and throat ward, emergency room, rehabilitation ward, cancer ward, and hospice ward. The results were as follows : 1. The highest scored items of the knowledge of pathophysiology, clinical microbiology, and mechanisms and effects of drugs necessary for nursing practice were side effects of drugs, anticoagulants, mechanisms of drugs, antihypertensive drugs, tolerance and addiction of drugs, interactions among drugs, hospital infection in the order of importance. The lowest scored item was structure of microorganisms. 2. The highest order of need according to unit was repair in tissue injury unit, definition etiology classification of inflammation in inflammation unit, transplantation and immunologic response in alterations in immunity unit, thrombus and thrombosis in disorders of cardiovascular function unit, gene disorders in genetic disorders unit, hospital infection in infection unit, virus in microorganisms unit, side reactions of drugs in introduction unit, anticonvulsants in drugs for central nervous system unit, local anesthesia in anesthesia unit, anticoagulants in drugs for cardiovascular system unit, anti-inflammatory drugs in antibiotics unit, anti-ulcer drugs in drugs for digestive system unit, and bronchodilators in drugs for respiratory system unit. 3. The common content of the knowledge of pathophysiology, clinical microbiology, and mechanisms and effects of drugs needed for all clinical areas in nursing were side effects of drugs, anticoagulants, interactions among drugs, and hospital infection. However, the degree of need of each pathological physiology, clinical microbiology, clinical microbiology, and mechanisms and effects of drugs was different depending on clinical areas. 4. Significant differences in the knowledge of pathophysiology, clinical microbiology, and mechanisms and effects of drugs necessary for nursing practice such as tissue changes due to injurious stimuli, degenerative changes of tissue, alterations in metabolism of carbohydrates, ischemia, hyperemia and congestion, hospital infection, structure of microorganism, classification of microorganism, bacteria, virus, antidepressants, antipsychotic drugs, antiemetic drugs, antiparkinsonism drugs, antianxiety drugs, antibiotics, tuberculostatics, antiviral drugs, antifungal drugs, parasiticides, antiulcer drugs, antidiarrheais, and anti constipation drugs were shown according to the work area. 5. Significant differences in the knowledge of pathophysiology, clinical microbiology, and mechanisms and effects of drugs necessary for nursing practice such as transplantation and immunologic response, alterations in the metabolism of uric acid, structure of microorganism, classification of microorganism, immunosuppressants, drugs for congestive heart failure were demonstrated according to the duration of work. Based on these findings, all the 72 items constructed by Korean Academic Society of Basic Nursing science should be included as contents of the knowledge of pathophysiology, clinical microbiology, and mechanisms and effects of drugs.

• Clinical and Experimental Pediatrics
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• v.45 no.9
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• pp.1106-1113
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• 2002

Purpose : Nuclear ($factor-{\kappa}BNF-{\kappa}B$) is now recognized as playing a potential role in programmed cell death and the adaptive response to various stress. Cellular hypoxia is a primary manifestation of many cardiovascular diseases. It seems that vascular endothelial growth factor (VEGF) and insulin like growth factor-I(IGF-I) have a function as a protective molecule in the heart against several stress including hypoxia. In this study, the role of $NF-{\kappa}B$ to the cellular response and regulation of protective molecules against the acute hypoxia in the heart was studied. Methods : To cause acute hypoxic stress to the heart, Sprague Dawley rats were exposed to hypoxic chamer($N_2$ 92% and $O_2$ 8%). After the hypoxic exposure, nuclear proteins, total proteins and mRNA were isolated from heart. Translocation of the transcription factors $NF-{\kappa}B$, NF-ATc, AP-1 and NKX-2.5 were evaluated by electrophoretic mobility shift assay(EMSA). The expression of IGF-I and VEGF were studied before and after the hypoxic stress by competitive-PCR, Northern hybridization and Western hybridization. To confirm the role of the $NF-{\kappa}B$ in the heart, the rats also were pretreated with diethyl-dithiocarbamic acid(DDTC) into peritoneal cavity to block $NF-{\kappa}B$ translocation into nucleus. Results : The expression of $NF-{\kappa}B$, AP-1 and NF-ATc were increased by the hypoxic stress. Increased expression of the VEGF and IGF-I were also observed by the hypoxic stress. However, the blocking of the $NF-{\kappa}B$ translocation reduced those expressions of VEGF and IGF-I. Conclusion : These results suggest that $NF-{\kappa}B$ has a protective role against the acute hypoxia through several gene expression, especially VEGF and IGF-I in heart muscle.

• Proceedings of the Microbiological Society of Korea Conference
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• 2008.05a
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• pp.39-41
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• 2008

The yeast Saccharomyces cerevisiae has been shown to contain three isoforms of citrate synthase (CS). The mitochondrial CS, Cit1, catalyzes the first reaction of the TCA cycle, i.e., condensation of acetyl-CoA and oxaloacetate to form citrate [1]. The peroxisomal CS, Cit2, participates in the glyoxylate cycle [2]. The third CS is a minor mitochondrial isofunctional enzyme, Cit3, and related to glycerol metabolism. However, the level of its intracellular activity is low and insufficient for metabolic needs of cells [3]. It has been reported that ${\Delta}cit1$ strain is not able to grow with acetate as a sole carbon source on either rich or minimal medium and that it shows a lag in attaining parental growth rates on nonfermentable carbon sources [2, 4, 5]. Cells of ${\Delta}cit2$, on the other hand, have similar growth phenotype as wild-type on various carbon sources. Thus, the biochemical basis of carbon metabolism in the yeast cells with deletion of CIT1 or CIT2 gene has not been clearly addressed yet. In the present study, we focused our efforts on understanding the function of Cit2 in utilizing $C_2$ carbon sources and then found that ${\Delta}cit1$ cells can grow on minimal medium containing $C_2$ carbon sources, such as acetate. We also analyzed that the characteristics of mutant strains defective in each of the genes encoding the enzymes involved in TCA and glyoxylate cycles and membrane carriers for metabolite transport. Our results suggest that citrate produced by peroxisomal CS can be utilized via glyoxylate cycle, and moreover that the glyoxylate cycle by itself functions as a fully competent metabolic pathway for acetate utilization in S. cerevisiae. We also studied the relationship between Cit1 and apoptosis in S. cerevisiae [6]. In multicellular organisms, apoptosis is a highly regulated process of cell death that allows a cell to self-degrade in order for the body to eliminate potentially threatening or undesired cells, and thus is a crucial event for common defense mechanisms and in development [7]. The process of cellular suicide is also present in unicellular organisms such as yeast Saccharomyces cerevisiae [8]. When unicellular organisms are exposed to harsh conditions, apoptosis may serve as a defense mechanism for the preservation of cell populations through the sacrifice of some members of a population to promote the survival of others [9]. Apoptosis in S. cerevisiae shows some typical features of mammalian apoptosis such as flipping of phosphatidylserine, membrane blebbing, chromatin condensation and margination, and DNA cleavage [10]. Yeast cells with ${\Delta}cit1$ deletion showed a temperature-sensitive growth phenotype, and displayed a rapid loss in viability associated with typical apoptotic hallmarks, i.e., ROS accumulation, nuclear fragmentation, DNA breakage, and phosphatidylserine translocation, when exposed to heat stress. Upon long-term cultivation, ${\Delta}cit1$ cells showed increased potentials for both aging-induced apoptosis and adaptive regrowth. Activation of the metacaspase Yca1 was detected during heat- or aging-induced apoptosis in ${\Delta}cit1$ cells, and accordingly, deletion of YCA1 suppressed the apoptotic phenotype caused by ${\Delta}cit1$ mutation. Cells with ${\Delta}cit1$ deletion showed higher tendency toward glutathione (GSH) depletion and subsequent ROS accumulation than the wild-type, which was rescued by exogenous GSH, glutamate, or glutathione disulfide (GSSG). Beside Cit1, other enzymes of TCA cycle and glutamate dehydrogenases (GDHs) were found to be involved in stress-induced apoptosis. Deletion of the genes encoding the TCA cycle enzymes and one of the three GDHs, Gdh3, caused increased sensitivity to heat stress. These results lead us to conclude that GSH deficiency in ${\Delta}cit1$ cells is caused by an insufficient supply of glutamate necessary for biosynthesis of GSH rather than the depletion of reducing power required for reduction of GSSG to GSH.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.415-422
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• 2005

library of the mutants was prepared by transposon mutagenesis of the Mycobacterium bovis BCG. We screened this library for the resistance to an anti-tuberculosis antibiotic, PA-824. Most of the mutants resistant to the PA-824 were not able to synthesize the coenzyme $F_{420}$ which is normally produced by the wild type M. bovis BCG strains. HPLC analysis of the cellular extract showed that one of those mutants which lost the ability to synthesize $F_{420}$ still produced F0. The insertion site of the transposon in this mutant was determined by an inverse PCR and the transposon was found to be inserted in the Rv2435c open reading frame (ORF). Rv2435c ORF is predicted to encode an 80.3 kDa protein. Rv2435c protein appears to be bound to the cytoplasmic membrane, its N-terminal present in the periplasm and C-terminal in the cytoplasm. The C-terminal portion of this protein is highly homologous with the adenylyl cyclases of both prokaryotes and eukaryotes. There are 15 ORFs which have homology with the class III AC proteins in the genome of the M. tuberculosis and M. bovis. Two of those, Rv1625c and Rv2435c, are highly homologous with the mammalian ACs. We cloned the cytoplasmic domain of the Rv2435c ORF and expressed it with six histidine residues attached on its C-terminal in Escherichia coli to find out if this protein is a genuine AC. Production of that protein in E. coli was proved by purifying the histidine-tagged protein by using the Ni-NTA resin. This protein, however, failed to complement the cya mutation in E. coli, indicating that this protein lacks the AC activity. All of the further attempts to convert this protein to a functional AC by a mutagenesis with UV or hydroxylamine, or construction of several different fusion proteins with Rv1625c failed. It is, therefore, possible that Rv2435c protein might affect the conversion of F0 to $F_{420}$ not by synthesizing cAMP but by some other way.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.43 no.2
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• pp.157-164
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• 2017

Stevia (Stevia rebaudiana) is a perennial plant of the genus Stevia, originated in South America. It stores many forms of glycosides, mainly stevioside and rebaudioside A, in which steviol is the basic structure. Steviol glycosides, widely used as sweeteners, are superior to sugar in sweetness. Recently, it has been reported that steviol glycosides are involved not only in the skin whitening and anti-inflammatory effect but also in enhancing skin barrier function through tight junction regulation. Thus, we examined anti-inflammatory effect of rebaudioside A and tried to identify its potential for improving atopic dermatitis as cosmetic ingredients. To investigate the anti-inflammatory effect, cell viability and mRNA expression level of inflammation-related cytokines were measured using mouse macrophage RAW264.7 cells. Cell counting kit 8 (CCK-8) assay was carried out to measure cell viability and the maximum concentration without cytotoxicity was set to $250{\mu}M$. A quantitative real-time RT-PCR method was used for the study of the inflammatory suppression of rebaudioside A. Rebaudioside A inhibited expression of inducible nitric oxide synthase (iNOS) up to 47% and COX-2 up to 41% compared to LPS treated condition. NO synthesis was decreased by rebaudioside A. Also, mRNA expression of interleukin (IL)-$1{\alpha}$, IL-$1{\beta}$ and IL-6 in LPS-stimulated RAW264.7 cells was decreased to 40%, 45% and 59%, respectively, as a concentration-dependent manner. In conclusion, rebaudioside A inhibited the inflammatory response by regulation of cytokine gene expression. From these results, we expect that steviol glycoside, such as rebaudioside A, can be used as a material for improving atopic dermatitis in the future.