• Journal of the Korean Applied Science and Technology
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• v.40 no.6
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• pp.1268-1277
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• 2023

In this study, to evaluate the possibility of utilizing Chamaecyparis obtusa (Siebold & Zucc.) Endl. (C. obtusa) leaf fractions as anti-inflammatory functional materials, C. obtusa extract extracted with 99% ethanol (CO99EL) was fractionated with hexane (CO99EL-H), chloroform (CO99EL-C), ethyl acetate (CO99EL-E), butanol (CO99EL-B) and distilled water (CO99EL-W). The anti-inflammatory effects of each fraction was performed using lipopolysaccharide (LPS)-induced RAW264.7 mouse macrophages. Cytotoxicity was highest in CO99EL-H and CO99EL-C and lowest in CO99EL-W. Interestingly, LPS-induced iNOS expression and NO production were significantly reduced by CO99EL-H and CO99EL-E, and COX-2 expression was significantly reduced by CO99EL-B and CO99EL-W. In addition, interleukin (IL)-1𝛽, an inflammatory cytokine increased by LPS, was significantly reduced by CO99EL-C, CO99EL-E, CO99EL-B and CO99EL-W, and IL-6 was significantly reduced by CO99EL-B and CO99EL-W. Therefore, the janus kinase (JAK)/signaling transducer and activator of transcription (STAT) signaling pathway activated by LPS was significantly reduced by CO99EL-H and CO99EL-C, and the mitogen-activated protein kinase (MAPK) signaling pathway was slightly reduced by CO99EL-H and CO99EL-C. However, nuclear factor (NF)-𝜅B activity was not reduced by any fractions. Based on the results of this study, it was confirmed that CO99EL fractions have different anti-inflammatory mechanisms depending on the solvent used for fractionation.

• Korean Journal of Microbiology
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• v.32 no.1
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• pp.47-52
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• 1994

The cells of Campylobacter jejuni heat-shocked at 48${\circ}C$ for 30 min synthesized the heat shock proteins of HSP90, HSP66 and HSP60. Those heat shock proteins were found to correspond to the heat shock proteins of HSP87, HSP66 (DnaK), and HSP58 (GroEL) of E. coli, respectively. By Southern blot analysis of the chromosomal DNAs of C. jejuni with groESL and dnaK genes of E. coli as DNA probes, the heat shock genes of C. jejuni which are homologous to the E. coli groESL and dnaK genes were found to exist in the chromosomal DNA. The genomic libraries of C. jejuni were constructed with the cosmid vector pWE15 and the groEL gene of C. jejuni were cloned in E. coli B178 groEL44 temperature senstive mutant. The hybrid plasmid (pLC1) was inserted with the DNA fragment (about 5.7kb in size) containing the groEL gene. E. coli groEL44 mutant cell transformed with the pLC1 could grow at 42${\circ}C$ by synthesizing the HSP60 of C. jejuni and regained the susceptibility to the ${\lambda}$ vir phage by expression of the groEL gene in the cloned cells. These indicated that the groEL products of C. jejuni had chaperon effects by synthesizing the heat shock proteins in the cloned cells of E. coli.

• Ocean and Polar Research
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• v.31 no.1
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• pp.51-61
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• 2009

In this study we define the two different types of El $Ni{\tilde{n}}o$, i.e., the eastern Pacific El $Ni{\tilde{n}}o$ (i.e., EP-El $Ni{\tilde{n}}o$) versus the central Pacific El $Ni{\tilde{n}}o$ (i.e., CP-El $Ni{\tilde{n}}o$), during the boreal summer (June-July-August, JJA) and winter (December-January-February, DJF) using the two NINO indices in the tropical Pacific. The two different types of El $Ni{\tilde{n}}o$ significantly differ in terms of the location of the maximum anomalous sea surface temperature (SST) in the tropical Pacific. The CP-El $Ni{\tilde{n}}o$ has been observed more frequently during recent decades compared to the EP-El $Ni{\tilde{n}}o$. In addition, our analysis indicates that the statistics of CP-El $Ni{\tilde{n}}o$ during JJA is closely associated with the warming trend in the central equatorial Pacific. We also examine the different responses of the East Asian marginal SST to the two types of El $Ni{\tilde{n}}o$ during JJA and DJF. The CP-El $Ni{\tilde{n}}o$ during both JJA and DJF is concurrent with warm SST anomalies around the Korean Peninsula including the East China Sea, which is in contrast to the EP-El $Ni{\tilde{n}}o$. Such different responses are associated with the difference in tropics/mid-latitude teleconnections via atmosphere between the two types of El $Ni{\tilde{n}}o$. Furthermore, our results indicate that atmospheric diabatic forcing in relation to the precipitation variability is different in the tropical Pacific between the EP-El $Ni{\tilde{n}}o$ and the CP-El $Ni{\tilde{n}}o$.

• Korean Journal of Veterinary Research
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• v.29 no.4
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• pp.469-474
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• 1989

The changes of plasma prostaglandin $E_2$ level, natural killer cell activity and tumor cell growth were assayed after transplantation of EL-4 leukemia cells in C57BL/6 mice. The results were summarized as follows; 1. Plasma prostaglandin $E_2$ level was increased in EL-4 bearing mice, but indomethacin treated mice group showed low level. 2. The tumor-derived prostaglandin $E_2$ inhibited the post-target binding cytolytic process of natural killer activity. 3. Indomethacin inhibited the growth of prostaglandin secreting EL-4 solid tumor.

• Journal of Microbiology
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• v.38 no.3
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• pp.145-149
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• 2000

GroEL is a typical molecular chaperone. GroEL synthesis patterns at various culture temperatures in Escherichia coli were investigated in this study. No significant differences in the amount of GroEL produced from the chromosome were found at 30 and 37$^{\circ}C$. However, GroEL production increased 3.4-fold at 42$^{\circ}C$. GroEL synthesis was not transient but continuous at 42$^{\circ}C$, although most heat shock gene expression is known to be transient. To understand the role of the groEL structural gene, a groE promoter-lacZ fusion was constructed. Interestingly , while transcriptional fusion is not thermally inducible, it is inducible by ethanol, suggesting that the secondary structure of the groEL transcript is involved in thermal regulation of the groEL gene. Secondary structures of groE mRNA at 37 and 42$^{\circ}C$ were compared using the computer program RNAdraw. Distinct structures at the two temperatures were found, and these structures may be related to a high level of GroEL expression at 42$^{\circ}C$.

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.337-346
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• 2016

Three combinations of molecular chaperones from Escherichia coli (i.e., DnaK-DnaJ-GrpE-GroEL-GroES, GroEL-GroES, and DnaK-DnaJ-GrpE) were overproduced in E. coli BL21, and their in vitro stabilizing effects on a nitrile hydratase (NHase) were assessed. The optimal gene combination, E. coli groEL-groES (ecgroEL-ES), was introduced into Rhodococcus ruber TH3. A novel engineered strain, R. ruber TH3G was constructed with the native NHase gene on its chromosome and the heterologous ecgroEL-ES genes in a shuttle plasmid. In R. ruber TH3G, NHase activity was enhanced 37.3% compared with the control, TH3. The in vivo stabilizing effect of ecGroEL-ES on the NHase was assessed using both acrylamide immersion and heat shock experiments. The inactivation behavior of the in vivo NHase after immersion in a solution of dynamically increased concentrations of acrylamide was particularly evident. When the acrylamide concentration was increased to 500 g/l (50%), the remaining NHase activity in TH3G was 38%, but in TH3, activity was reduced to 10%. Reactivation of the in vivo NHases after varying degrees of inactivation was further assessed. The activity of the reactivated NHase was more than 2-fold greater in TH3G than in TH3. The hydration synthesis of acrylamide catalyzed by the in vivo NHase was performed with continuous acrylonitrile feeding. The final concentration of acrylamide was 640 g/l when catalyzed by TH3G, compared with 490 g/l acrylamide by TH3. This study is the first to show that the chaperones ecGroEL-ES work well in Rhodococcus and simultaneously possess protein-folding assistance functions and the ability to stabilize and reactivate the native NHases.

• Archives of Pharmacal Research
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• v.17 no.3
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• pp.194-198
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• 1994

The antitumor effects of the 2E4 and anti--Tac, monoclonal antibodies directed to Il-2 receptor (IL-2R) conjugated with .alpha.-particle emitting radionuclide bismuth-212., were compared. The $^{212}Bi-2E4$ demonstratedspecific cytotocicity to EL4J3, 4, a I$L-2R^+$ cell line, than to EL4J, a $IL-2R^-$ cell line in thymidine incorporation assy. TEX>$^{212}Bi-2E4$ exerted the maximal antitumor effect in that % T/C in C57BL/6 mice implanted with EL4J3.4 ascitic tumor was 331% at the concentration of $50{\;}{\mu}Ci$, while that of $^{212}Bi-anti-Tac$ was 258% at $100{\;}{\mu}Ci$.

• Biomedical Science Letters
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• v.4 no.2
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• pp.129-135
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• 1998

We have carried out polymerase chain reaction (PCR) to investigate if retropseudogene for CYP2El is present in human genome. PCR primers were designed based on the structure of functional CYP2El gene and used to amplify both functional gene and retropseudogene in one reaction. From the repeated experiments we were able to amplify a previously unidentified CYP2El retropseudogene that was present in human genome. Its detailed structure was confirmed by Southern blotting and DNA sequencing. Nucleotide sequence of this retropseudogene was completely matched up to human liver CYP2El mRNA suggesting that the development of this retropseudogene might be a relatively recent event.

• KSBB Journal
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• v.14 no.4
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• pp.429-433
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• 1999

$\beta$-Glucosidaes from Cellvibrio gilvus(CG) was successfully overproduced in soluble form in E. coli with the coexpression of GroEL/ES/. Without the GroEL/ES protein, the $\beta$-glucosidase overexpressed in E. coli constituted a huge amount(80%) of total cellular protein, but was localized in the insoluble fraction, and little activity was detected in the soluble fraction. Coexpression of the E. coli GroEL/ES had a drastic impact on the proper folding of the $\beta$-glucosidase; 20% of the overexpressed enzyme was recovered in the soluble fraction in active form. Similar effects of GroEL/ES were also observed on the overexpressed $\beta$-glucosidase from Agrobacterium tumefaciens(AT). And pET28(a)-RGRAR, partially deleted mutant lacking 5-amino acid residues at carboxy teminus also could be folded into an active form when expressed with the molecular chaperonin GroEL/ES, and its activity was higher than that of the without GroEL/ES system, In addition, the synergistic effect of GroEL/ES and the low induction temperature were important factors for solubilization of the inclusion body from overproduced $\beta$-glucosidases.

• Journal of Life Science
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• v.12 no.6
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• pp.688-693
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• 2002

Molecular chaperones prevent the misfolding of newly synthesized polypeptides in the cell. The coexpression of molecular chaperones could be expected to improve the production of soluble and active recombinant proteins. In this study, the effect of coexpression of E. coli GroEL/ES chaperone on the active production of Bacillus macerans cyclodextrin glucanotransferase (CGTase) in E. coli was investigated. Two plasmids, pTCGT1 and pGro7 in which the cgt and the groEL/ES genes are under the control of 77 promoter and araB promoter, respectively, were co-transformed into E. coli. With a series of cultures of recombinant E. coli cells, the optimal concentrations of IPTG and L-arabinose were found be 1 mM and 0.3 mg/$m\ell$, respectively. When IPTG and L-arabinose were added at 0.8~1.0 $OD_{600}$ and 0.4~0.5 $OD_{600}$, active CGTase production was increased significantly. This coexpression condition resulted in 1.5-fold increased level of soluble CGTase (0.7~0.73 unit/$m\ell$), compared to the level of CGTase in the single expression (0.36~0.56 unit/$m\ell$). An SDS-PACE analysis revealed that about 33.6% of CGTase in the total CGTase protein was found in the soluble fraction by coexpression of GroEL/ES chaperone.