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Immunomodulating Activity of the Exopolymer from Submerged Mycelial Culture of Phellinus pini

  • Jeong, Sang-Chul;Cho, Sung-Pill;Yang, Byung-Keun;Jeong, Yong-Tae;Ra, Kyung-Soo;Song, Chi-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.15-21
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    • 2004
  • The immunomodulating activities and chemical characteristics of a water-soluble exopolymer from submerged mycelial culture of Phellinus pini were studied. Anticomplementary activity of this polymer was found to be $73.2\%$, and its activation system occurred through both classical and alternative pathways, where the classical pathway was detected to be the major one by crossed immunoelectrophoresis. Nitric oxide (NO) release ability and acid phosphatase activity of macrophage were increased by 1.6-fold ($100{\mu}g/ml$) and 3.4-fold ($500{\mu}g/ml$), respectively, and splenocyte proliferation in mixed lymphocyte reaction (MLR) was also increased by 2.6-fold ($200{\mu}g/ml$), compared to the control. The molecular weight of this polymer, determined by HPLC, was under 5 kDa. Total sugar and protein contents were 89.7 and 10.3%, respectively. Both sugar and amino acid compositions of the exopolymer were also analyzed.

Overexpression of Arylsulfatase in E. coli and Its Application to Desulfatation of Agar

  • Lim, Jae-Myung;Jang, Yeon-Hwa;Kim, Hyeung-Rak;Kim, Young-Tae;Choi, Tae-Jin;Kim, Joong-Kyun;Nam, Soo-Wan
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.777-782
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    • 2004
  • The arylsulfatase gene (astA, 984 bp ORF) from the P. carrageenovora genome was amplified by PCR and subcloned into the pET21a vector. When the constructed plasmid pAST-A1 (6.4 kb) was introduced into E. coli BL21(DE3), the transformant on the LB plate containing IPTG showed a hydrolyzing activity for 4-methylumbelliferyl sulfate and p-nitrophenyl sulfate. The highest arylsulfatase activity (2.1 unit/ml) was obtained at 10 mM IPTG. Most arylsulfatase activity was found in the cell lysate, whereas no significant activity was detected in the culture supernatant. The molecular weight of the recombinant enzyme was estimated to be 33.1 kDa by SDS-PAGE. After the reaction of agar with arylsulfatase for 12 h at $40^{\circ}C$, the gel strength of the agar increased by 2-fold, and 73% of the sulfate in the agar had been removed. This result suggests that arylsulfatase expressed in E. coli could be useful in the production of electrophoretic grade agarose.

A Toxicogenomic Study to Assess Methylmercury-induced Neurotoxicity

  • Kim, Youn-Jung;Yun, Hye-Jung;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.10a
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    • pp.177-177
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    • 2003
  • Methylmercury (MeHg) is a well-known neurotoxicant that causes severe damage to the central nervous system in humans. Many reports have shown that MeHg is poisonous to human body through contaminated foods and has released into the environment. Despite many studies on the pathogenesis of MeHg-induced central neuropathy, no useful mechanism of toxicity has been established so far. In this study, suppressive subtractive hybridization (SSH) was performed to identify differentially expressed genes on human neuroblastoma cell line, SH-SY5Y treated with DMSO and MeHg (6.25 uM) for 6 hr. Differentially expressed cDNA clones were sequenced and were screened by dot blot to eliminate false positive clones. 13 of 35 screened genes were confirmed using real time RT-PCR. These genes include EB1,90-kDa heat-shock protein, chromosome condensation-related SMC-associated protein and brain peptide Al, etc. Analysis of these genes may provide an insight into the neurotoxic effects of MeHg in human neuronal cells and a possibility to develop more efficient and exact monitoring system of heavy metals as ubiquitous environmental pollutants.

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Centrobin/Nip2 Expression In Vivo Suggests Its Involvement in Cell Proliferation

  • Lee, Jungmin;Kim, Sunmi;Jeong, Yeontae;Rhee, Kunsoo
    • Molecules and Cells
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    • v.28 no.1
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    • pp.31-36
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    • 2009
  • Centrobin/Nip2 was initially identified as a centrosome protein that is critical for centrosome duplication and spindle assembly. In the present study, we determined the expression and subcellular localization of centrobin in selected mouse tissues. Immunoblot analysis revealed that the centrobin-specific band of 100 kDa was detected in all tissues tested but most abundantly in the thymus, spleen and testis. In the testis, centrobin was localized at the centrosomes of spermatocytes and early round spermatids, but no specific signal was detected in late round spermatids and elongated spermatids. Our results also revealed that the centrosome duplication occurs at interphase of the second meiotic division of the mouse male germ cells. The centrobin protein was more abundant in the mitotically active ovarian follicular cells and thymic cortex cells than in non-proliferating corpus luteal cells and thymic medullary cells. The expression pattern of centrobin suggests that the biological functions of centrobin are related to cell proliferation. Consistent with the proposal, we observed reduction of the centrobin levels when NIH3T3 became quiescent in the serum-starved culture conditions. However, a residual amount of centrobin was also detected at the centrosomes of the resting cells, suggesting its role for maintaining integrity of the centrosome, especially of the daughter centriole in the cells.

Effect of Weight Shift Training with Electrical Sensory Stimulation Feedback on Standing Balance in Stroke patients

  • Kim, Da-young;Cha, Yong-jun
    • Journal of the Korean Society of Physical Medicine
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    • v.10 no.3
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    • pp.39-45
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    • 2015
  • PURPOSE: The purpose of this study was to investigate the effect of weight shift training with electrical sensory simulation feedback on quiet standing balance in hemiplegic stroke patients. METHODS: 30 stroke patients were equally allocated at random to an experimental group or a control group. Patients in both two groups underwent comprehensive rehabilitation physical therapy for 30 minutes per day for 5 days per week for 4 weeks. Members of the experimental group received additional weight shift training with electrical sensory simulation feedback was conducted for 15 minutes after 30 minute sessions, whereas members of the control group underwent additional leftward/rightward weight shift training by themselves after 30 minutes per day for four weeks. COP (center of pressure) path lengths, COP velocities, and foot forces were measured before and immediately after the 4-week training period in both groups and results were compared. RESULTS: COP path lengths significantly decreased by 3% after training in the experimental group and this was significantly greater than that observed in the control group (p<0.05). In both groups, foot forces of affected sides showed significant increases after intervention, whereas foot forces of unaffected sides showed significant decreases (p<0.05). No significant difference was observed between the two groups with respect to these changes. CONCLUSION: Weight shift training using electrical sensory simulation feedback has a positive effect on quiet standing balance in hemiplegic stroke.

Internal Consistency and Concurrent Validity of Korean Language Version of WHODAS 2.0: 12 Item-Self Administered

  • Lee, Hae-Jung;Kim, Da-Jeong
    • The Journal of Korean Physical Therapy
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    • v.23 no.6
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    • pp.23-29
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    • 2011
  • Purpose: The aim of this study was to validate the Korean version of World Health Organization Disability Assessment Schedule 2.0 (KWHODAS 2.0) in 12 item-self administered version (12-self). Methods: The KWHODAS 2.0 and Korean Functional Rating index (KFRI) were tested for internal consistency, ceiling and floor effects, and concurrent validity in 111 patients with low back pain and/or neck pain. Results: A very high level of internal consistency was shown for both instruments; ${\alpha}$=0.96 with KWHODAS 2.0; 12-self and ${\alpha}$=0.97 for KFRI. No ceiling and/or floor effects were found in both the instruments. The KWHODAS 2.0 and KFRI were highly correlated (r=0.77), and the relationship of each item between KWHODAS 2.0 and KFRI was ranging from r=0.09 to 0.72. Conclusion: We conclude that the KWHODAS 2.0: 12-self and KFRI are reliable and are valid instruments for the measurement of disability in Korean speaking patients with low back and/or neck pain. Both instruments, the KWHODAS 2.0; 12-Self and KFRI are now suitable for use in clinical practice and research applications.

Growth Inhibitory Effect of Fermented Kimchi on Food-borne Pathogens

  • Lee, Jong-Kyung;Jung, Da-Wa;Kim, Yun-Ji;Cha, Seong-Kwan;Lee, Myung-Ki;Ahn, Byung-Hak;Kwak, No-Seong;Oh, Se-Wook
    • Food Science and Biotechnology
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    • v.18 no.1
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    • pp.12-17
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    • 2009
  • The effect of kimchi, traditional Korean fermented vegetables, on inactivating food-borne pathogens and the kimchi factors affecting the antimicrobial activity were investigated. More cells of Listeria monocytogenes, Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella typhimurium were inactivated in the kimchi that had low pH and high titratable acidity. Of the raw ingredients in kimchi, raw garlic showed the strongest antimicrobial activity against the pathogens. When kimchi was fermented at 0, 4, 10, or $20^{\circ}C$ to pH 4.4, higher kimchi fermentation temperature resulted in higher titratable acidity. The greatest inactivation of S. typhimurium occurred in kimchi fermented at $20^{\circ}C$, while L. monocytogenes were inactivated in kimchi fermented at $0^{\circ}C$ in situ. This study showed that appropriately fermented kimchi can inactivate various food-borne pathogens and that the fermentation temperature of the kimchi is an important factor in determining the ability of the kimchi to inactivate specific pathogens. Lactic acid bacteria (LAB) multiplication and organic acids produced according to LAB metabolism play a role in inactivating food-borne pathogens in kimchi.

Allergenicity of Hot Peppers Cultivated in Korea

  • Lee, Jeong-Ok;Kim, Eun-Jung;Ko, Yu-Jin;Lee, Sang-Il;Lee, Won-Sup;Ryu, Chung-Ho
    • Food Science and Biotechnology
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    • v.18 no.2
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    • pp.317-322
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    • 2009
  • The proteins from 15 types of cultivar of hot peppers cultivated in Korea were extracted and its allergenicity was investigated by immunoblotting and enzyme-linked immunosorbent assay (ELISA). The immunoblotting of hot pepper proteins extracts (HPEs) against serum of hot pepper sensitized patients revealed dominant IgE binding to 14, 37, and 40 kDa molecules. The specific levels of IgE to HPEs sample No. 1, 3, and 7 were much higher than the other samples in patients. Also, IgE binding capacity of HPEs were not reduced by thermal processing and digestion in ELISA using human IgE antibody acquired from hot pepper sensitized patients. By means of Western blotting using anti-thaumatin IgY, thaumatin-like protein (TLP) acting as allergen in several plants and fruits was detected in tested hot peppers. This study demonstrates that the antigenic protein in hot peppers are present but are differently contained according to cultivars.

Detection of Fish Virus by Using Immunomagnetic Separation and Polymerase Chain Reaction (IMS-PCR)

  • KIM Soo Jin;OH Hae Keun;CHOI Tae-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.6
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    • pp.948-955
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    • 1997
  • Immunomagnetic separation of virus coupled with .reverse transcription-polymerase chain reaction (IMS-PCR) was performed with infectious hematopoietic necrosis virus (IHNV). A DNA fragment of expected size was synthesized in the RT-PCR with total RNA extracted from IHNV inoculated CHSE-214. In a SDS-PAGE analysis, a protein band of over 70kDa was detected from non-infected cells and cells inoculated with IHNV and infectious pancreatic necrosis virus (IPNV). This protein was detected in the Western blot analysis probably because of non-specific reaction to monoclonal antibody against IHNV nucleocapsid protein. In the immunomagnetic separation, magnetic beads coated with monoclonal antibody against the IHNV nucleocapsid protein was incubated with supernatant from IHNV inoculated CHSE-214 cells. During this process, the non-specifically reacting protein could be removed by washing the magnetic bead with PBS in the presence of an external magnetic field, and viral proteins were detected from the remaining, cleaned magnetic beads. It was necessary to extract viral RNA from the captured virus particles before RT-PCR, and no DNA product was detected when the captured virus was only heated 5 min at $95^{\circ}C$. A PCR-product of expected size was synthesized from IMS-PCR with magnetic beads double coated either by goat anti-mouse IgG antibody -monoclonal antibody or streptavidin - biotin conjugated monoclonal antibody.

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