Proceedings of the Korean Society of Plant Biotechnology Conference
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2003.04a
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pp.65-71
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2003
Bax, a mammalian pro-apoptotic member of the Bcl-2 family, induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorho-damine 123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MBP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to play a novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.
The residential and welfare facilities for the elderly are continuously increasing due to change of value on family under situation of aging population increase. And it is predicted that accommodation capacity of facilities for the elderly and its rate those facility takes compared to whole social welfare facilities will be accelerated considering past increase speed. On the other hand, about 60% of elderly people have low physical and mental level almost dose to handicapped people therefore special environmental concerns helping their independent living are necessary. The purpose of this study is to analyze whether facilities for the elderly are adequate to accommodate their request condition by understanding color among environmental factors is one of most important factor for smooth understanding, communication and psychological remedy effect for thou. For this purpose, importance and effect of color and visual characteristic and reaction to color in elderly environment are researched through documents and visited 10 facilities in Seoul and Kyunggi region to research interior color status. And measuring of color on 5 main spaces of the facilities such as lobby/lounge, corridor, dining room, bedroom, stairway/ramp are done under analysis of its functional and aesthetic level based on Moon & Spencer's color theory.
The purposes of this study is to discuss and analyze the effect on the recovery from cut in sciatic nerve. This study used 9 weeks male rats of Sprague-Dawley family. Rat in groups 4 were treated with pulsed therapeutic ultrasound for 3 minutes. 3 times weekly at 3MHz respectively (intensity; $0.2W/cm^2,\;0.5W/Cm^2,\;10W/cm^2$); rat in group 1 received placebo ultrasound. In addition, changes of serum aspartate amino-transferase(AST) and creatine phosphokinase(CPK) levels were also demonstrated with diameter of individual muscle fasciculate and number of muscle fiber in each of three types of muscles located in gastrocnemius, soles. The results of comparing the changes in groups are as follows; 1. We found out that hypertrophic epineurium was present in sciatic nerve injured ultrasound treatment of groups. 2. In the gastrocnemius morphological investigation of the group I (control group), severe muscle atrophy were observed at the 7th days of the sciatic nerve injury. however, muscle atrophy of the group IV ($1.0W/cm^2$) were slightly recovered at the 14th days after treatment ultrasound. At the 28th days, muscular fibers were formed in polygon and were significantly recovered. 3. C-fos immunoreactive of the group II ($0.2W/cm^2$), III ($0.5W/cm^2$) were remarkably increased at the 1th day after treatment of ultrasound. Group IV were markedly deceased. 4. Brain-Derived Neurotrophic Factor(BDNF) immunoreactive of the group II, III were increased after 7 days of the sciatic nerve injury. Group IV were markedly increased from 14th days to 28th days after treatment of ultrasound. 5. A significant increase of serum AST levels were demonstrated in control group. However, serum AST levels of massage groups were significantly decreased compared to that of control group in followed order ; ($0.2W/cm^2<0.5W/cm^2<1.0W/cm^2$). 6. A significant increase of serum CK levels were demonstrated in control of group. However, serum CK levels of massage groups were significantly decreased compared to that of control group in followed order ; ($0.2W/cm^2<0.5W/cm^2<1.0W/cm^2$). The above results suggest that ultrasound treatment after peripheral nerve injury might reduce noxious stimuli, facilitate nerve recovery and effective in the functional improvement delaying muscle atrophy or degeneration.
A putative cyclomaltodextrinase gene (licd) was found from the genome of Listeria innocua ATCC 33090. The licd gene is located in the gene cluster involved in maltose/maltodextrin utilization, which consists of various genes encoding maltose phosphorylase and sugar ABC transporters. The structural gene encodes 591 amino acids with a predicted molecular mass of 68.6 kDa, which shares less than 58% of amino acid sequence identity with other known CDase family enzymes. The licd gene was cloned, and the dimeric enzyme with C-terminal six-histidines was successfully produced and purified from recombinant Escherichia coli. The enzyme showed the highest activity at pH 7.0 and 37℃. licd could hydrolyze β-cyclodextrin, starch, and maltotriose to mainly maltose, and it cleaved pullulan to panose. It could also catalyze the hydrolysis of acarbose to glucose and acarviosine-glucose. In particular, it showed significantly higher activity towards β-cyclodextrin and maltotriose than towards starch and acarbose. licd also showed transglycosylation activity, producing α-(1,6)- and/or α-(1,3)-linked transfer products from the acarbose donor and α-methyl glucopyranoside acceptor.
An isoflavone glucosidase that catalyzes the hydrolysis of isoflavone glucosides into glucose and corresponding aglycones was purified from Candida fermentati SI. The N-terminal sequence was determined to be GLNCDYCN. We designed degenerate primers on the basis of these amino acid sequences and successfully cloned the full structural gene sequence of the isoflavone glucosidase using inverse PCR. The exo-β-(1,3)-glucanase gene consists of 1227 base-pair nucleotides, encoding a 408-amino-acid sequence that shares 41–96% amino acid homology with other yeast exo-β-(1,3)-glucanases belonging to glycoside hydrolase family 5. The recombinant exo-β-(1,3)-glucanase was expressed in Pichia pastoris X-33, using a pPICZA vector system, and further characterized. The molecular mass of the purified exo-β-(1,3)-glucanase was estimated by SDS-PAGE to be 47 kDa. The optimal pH and temperature were pH 4.5 and 40℃, respectively. The Km values of the purified exo-β-(1,3)-glucanase for daidzin and genistin were 0.12 mM and 0.14 mM, respectively. The Vmax values of the purified isoflavone glucosidase were 945.03 U/mg for daidzin and 835.92 U/mg and for genistin.
The oxidation ditch (OD) is one of the most widely used processes for treating municipal wastewater. However, the microbial communities in the OD systems have not been well characterized, and little information about the shift of bacterial community during the startup process of the OD systems is available. In this study, we investigated the bacterial community changes during the startup period (over 100 days) of a full-scale OD. The results showed that the bacterial community dramatically changed during the startup period. Similar to the activated sludge samples in other studies, Proteobacteria (accounting for 26.3%-48.4%) was the most dominant bacterial phylum in the OD system, but its relative abundance declined nearly 40% during the startup process. It was also found that Planctomycetes proliferated greatly (from 4.79% to 13.5%) and finally replaced Bacteroidetes as the second abundant phylum in the OD system. Specifically, some bacteria affiliated with genus Flavobacterium exhibited remarkable decreasing trends, whereas bacterial species belonging to the OD1 candidate division and Saprospiraceae family were found to increase during the startup process. Despite of the bacterial community shift, the organic matter, nitrogen, and phosphorus in the effluent were always in low concentrations, suggesting the functional redundancy of the bacterial community. Moreover, by comparing with the bacterial community in other municipal wastewater treatment bioreactors, some potentially novel bacterial species were found to be present in the OD system. Collectively, this study improved our understandings of the bacterial community structure and microbial ecology during the startup of a full-scale wastewater treatment bioreactor.
Lee, Ran;Park, Hyun Jung;Lee, Won Young;Kim, Ji Hyuk;Kim, In Chul;Kim, Dong Woon;Lee, Sung Dae;Jung, Hyun Jung;Kim, Jong Moon;Yoon, Hyung Moon;Kwon, Hyuk Jung;Song, Hyuk
Reproductive and Developmental Biology
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v.36
no.3
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pp.225-230
/
2012
Interferon induced transmembrane protein-1 (Ifitm-1) has been reported to have an important role in primordial germ cell formation, and it has expressed in female reproductive organ. In the present study, Ifitm-1 gene expression was identified in testes and all part of epididymis using western immunoblot and immunohistochemistry. Interestingly, Ifitm-1 expression was observed on the head of spermatozoa. To investigate the role of Ifitm-1 gene expression in behavior of spermatozoa after acrosome reaction, fresh sperm was incubated with calcium ionophore to induce acrosome reaction, whereas the expression of Ifitm-1 was not altered after the acrosome reaction. Then to identify the effect of Ifitm-1 in sperm motility and other seminal parameters, different concentration of Ifitm-1 antibody was incubated with spermatozoa, and seminal parameters were assessed using computer-assisted semen analysis (CASA). Interestingly, motility, progressive, and VAP were increased in the sperm with Ifitm-1 antibody treated compared to rabbit serum, however other parameters such as straightness were not changed. In order to identify the functional significance of Ifitm-1 in fertilization, capacitated spermatozoa were pre-incubated with anti-Ifitm-1 antibody and subsequently examined the ability to adhere to mouse oocytes. However, any defection or alteration in sperm-egg fusion was not found, Ifitm-1 antibody treated or non-treated spermatozoa showed a normal penetration. Although the precise role of Ifitm-1 in sperm motility and following fertilization need to be elucidated, this study suggests that the activation of Ifitm-1 on the sperm may enhance the motility of spermatozoa in mice.
Journal of the Korea Academia-Industrial cooperation Society
/
v.21
no.7
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pp.656-666
/
2020
This study was a descriptive research study for hemodialysis patients to survey the effects of social support and health literacy on treatment adherence. The subjects of this study were 140 hemodialysis patients aged over 40 years who had been receiving treatment for more 1 year in artificial kidney rooms at two general hospitals in Y city. The data were collected from November 1, 2019 to December 31, 2019 and were analyzed using descriptive statistics, t-tests, one-way ANOVA, Scheffe test, Pearson's correlation coefficients, and multiple regression with the SPSS/WIN 26.0 statistical program. The results of this study show that social support (family, friends, medical staff) and health literacy (functional, communication, critical) were positively correlated with treatment adherence. The variables affecting treatment adherence in hemodialysis patients were identified by social support and health literacy with 69.6% explanatory power. To improve the treatment adherence of hemodialysis patients, it is necessary to develop education programs to improve health literacy based on social support.
Kim, Yeon-Kye;Moon, Ho-Sung;Lee, Moon-Hee;Park, Mi-Ju;Lim, Chi-Won;Park, Hee-Yeon;Park, Jin-Il;Yoon, Ho-Dong;Kim, Dae-Hee
Korean Journal of Fisheries and Aquatic Sciences
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v.42
no.5
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pp.434-441
/
2009
This study was conducted to compare the biological activities of 7 melania snails from the family Pleuroceridae (Semisulcospira coreana, Koreanomelania nodifila, Semisulcospira forticosta, Koreoleptoxis globus ovalis, Semisulcospira libertina, Semisulcospira tegulata and Semisulcospira gottschei) in Korea. Among the 7 species, S. coreana, Korean. nodifila, S. forticosta and S. gottschei showed over 80% cytotoxicities on three cancer cell lines (SNU-1, A549 and Hep 3B) compared to the non-treatment, whereas S. libertina and S. tegulata showed almost no growth inhibition activities on the same cancer cell lines. In relation to ACE inhibition activity, only S. coreana, Korean. nodifila, and S. forticosta showed over 60% ACE inhibition activities, whereas other melania snails exhibited inhibition activities of lower than 25%. DPPH radical scavenging activities were also determined, and used to categories melania snails into three groups based on Duncan's multiple range test at P<0.05. The amount of TNF-${\alpha}$ produced by in vitro mouse peritoneal macrophage was determined according to bioactivity on L-929 cells. Three melania snails, S. coreana, Korean. nodifila and S. gottschei, exhibited 95.2%, 89.7% and 93.7% cell death(%) on L-929 cells, respectively. Glucose-6-phosphate dehydrogenase inhibitory activity was also obtained in the extract of S. coreana (31.9%) and Korean. nodifila (28.1%), showing that these extracts can be used as supplemental dietary health foods. In conclusion, we believe that the extracts of melania snails should be given due consideration in functional health food development.
Activation of c-Jun N-terminal kinase (JNK), a member of the mitogen-activated protein kinase family, is an important cellular response that modulates the outcome of the cells which are exposed to the tumor necrosis factor (TNF) or the genotoxic stress including DNA damaging agents. Although it is known that JNK is activated in response to genotoxic stress, neither the pathways to transduce signals to activate JNK nor the primary sensors of the cells that trigger the stress response have been identified. Here, we report that the receptor interacting protein (RIP), a key adaptor protein of TNF signaling, was required to activate JNK in the cells treated with certain DNA damaging agents such as adriamycin (Adr) and 1-${\beta}$-D-arabinofuranosylcytosine (Ara-C) that cause slow and sustained activation, but it was not required when treated with N-methyl-N-nitro-N-nitrosoguanidine (MNNG) and short wavelength UV, which causes quick and transient activation. Our findings revealed that this sustained JNK activation was not mediated by the TNF (tumor necrosis factor) receptor signaling, but it required a functional ATM (ataxia telangiectasia) activity. In addition, JNK inhibitor SP-600125 significantly blocked the Adr-induced cell death, but it did not affect the cell death induced by MNNG. These findings suggest that the sustained activation of JNK mediated by RIP plays an important role in the DNA damage-induced cell death, and that the duration of JNK activation relays a different stress response to determine the cell fate.
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