• Biomolecules & Therapeutics
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• 제11권3호
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• pp.157-162
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• 2003

The generation of secretory IgA antibodies(Abs) for specific immune protection of mucosal surfaces depends on stimulation of the mucosal immune system, but this is not effectively achieved by parenteral or even oral administration of most soluble antigens. Thus, to produce a possible vaccine antigen against urinary tract infections, the uropathogenic E. coli (UPEC) adhesin was genetically coupled to the ctxa2b gene and cloned into a pMAL-p2E expression vector. The chimeric construction of pMALfimHIctxa2b was then transformed into E. coli K-12 TB1 and its nucleotide sequence was verified. The chimeric protein was then purified by applying the affinity chromatography. The purified chimeric protein was confirmed by SDS-PAGE and western blotting using antibodies to the maltose binding protein (MBP) or the cholera toxin subunit B (CTXB), plus the N-terminal amino acid sequence was analyzed. The orderly-assembled chimeric protein was confirmed by a modified $G_{M1}$-ganglioside ELISA using antibodies to adhesin. The results indicate that the purified chimeric protein was an Adhesin/CTXA2B protein containing UPEC adhesin and the $G_{M1}$-ganglioside binding activity of CTXB. This study also demonstrate that peroral administration of this chimeric immunogen in mice elicited high level of secretory IgA and serum IgG Abs to the UPEC adhesin. The results suggest that the genetically linked CTXA2B acts as a useful mucosal adjuvant, and that the adhesin/CTXA2B chimeric protein might be a potential antigen for oral immunization against UPEC.

• BMB Reports
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• 제52권6호
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• pp.397-402
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• 2019

Middle East respiratory syndrome coronavirus (MERS-CoV) uses the spike (S) glycoprotein to recognize and enter target cells. In this study, we selected two epitope peptide sequences within the receptor binding domain (RBD) of the MERS-CoV S protein. We used a complex consisting of the epitope peptide of the MERS-CoV S protein and CpG-DNA encapsulated in liposome complex to immunize mice, and produced the monoclonal antibodies 506-2G10G5 and 492-1G10E4E2. The western blotting data showed that both monoclonal antibodies detected the S protein and immunoprecipitated the native form of the S protein. Indirect immunofluorescence and confocal analysis suggested strong reactivity of the antibodies towards the S protein of MERS-CoV virus infected Vero cells. Furthermore, the 506-2G10G5 monoclonal antibody significantly reduced plaque formation in MERS-CoV infected Vero cells compared to normal mouse IgG and 492-1G10E4E2. Thus, we successfully produced a monoclonal antibody directed against the RBD domain of the S protein which could be used in the development of diagnostics and therapeutic applications in the future.

• 한국수산과학회지
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• 제54권1호
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• pp.118-123
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• 2021

This study was conducted to evaluate plant proteins as a replacement for a fishmeal diet in the rearing of coho salmon Oncorhynchus kisutch. Twelve groups of 20 fish averaging 34.0±0.62 g were randomly distributed into 12 rectangular tanks (250 L). Four experimental diets included a control diet containing 60% fishmeal (Control), and three other diets that replaced 20% of fishmeal with soy protein concentrate (SPC), fermented soybean protein concentrate (F-SPC), and enzyme-processed soy protein concentrate (E-SPC). At the end of the feeding trial, fish that were fed Control, SPC and E-SPC diets showed significantly higher weight gain, specific growth rate, feed efficiency, and protein efficiency ratio than those that were fed F-SPC diet. However, there were no significant differences among the fish that were fed Control, SPC, and E-SPC diets. No significant differences were observed in crude protein, crude lipid, and ash of whole body among the fish that were fed all the diets. Therefore, these results indicated that 20% of fishmeal could be replaced by E-SPC or SPC without any adverse effects on the growth performance of coho salmon.

• 대한한의학회지
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• 제43권3호
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• pp.106-121
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• 2022

Objectives: The purpose of this study was to investigate the anti-inflammatory effect of Cornus Officinalis fruit extract(CE) and Cornus Officinalis Fruit Cheonghyeol Plus(CCP) in Human Umbilical Vein Endothelial Cell. Methods: We measured cell viability of CE, CCP and treated HUVEC with TNF-α. We measured the mRNA expression levels of KLF2, eNOS, MCP-1, ICAM-1, VCAM-1, the protein expression levels of KLF2, eNOS, MCP-1, ICAM-1, VCAM-1, and the protein phosphorylation level of ERK, JNK, p38 and the biomarker expression levels of MCP-1, ICAM-1, VCAM-1. Results: 1.CE incresed the mRNA, protein expression levels of KLF2, eNOS at concentrations of 100㎍/㎖ compared to the control group. CE decresed the mRNA, protein and biomarker expression levels of MCP-1,ICAM-1,VCAM-1 at concentrations of 100㎍/㎖ compared to the control group. CE decresed the protein phosphorylation level of p38 at concentrations of 100㎍/㎖ compared to the control group. 2. CCP incresed the mRNA, protein expression levels of KLF2, eNOS at concentrations of 100㎍/㎖ or more compared to the control group. CCP decresed the mRNA, protein and biomarker expression levels of MCP-1, ICAM-1, VCAM-1 at concentrations of 100㎍/㎖ or more compared to the control group. CCP decresed the protein phosphorylation level of ERK at concentrations of 100㎍/㎖ or more, p38 at concentrations of 200㎍/㎖ or more, and JNK at concentrations of 400㎍/㎖ compared to the control group. Conclusions: These results present that CE and CCP has anti-inflammatory effect in HUVEC. So, it could help treat or prevent inflammation in vein caused by dyslipidemia and contribute prevention of cardiovascular and cerebrovascular cerebrovascular diseases.

• Archives of Pharmacal Research
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• 제23권4호
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• pp.267-282
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• 2000

Cytochrome P45O (CYP) 2E1 catalyzes the metabolism of a wide variety of therapeutic agents, procarcinogens, and low molecular weight solvents. CYP2E1-catalyzed metabolism may cause toxicity or DNA damage through the production of toxic metabolites, oxygen radicals, and lipid peroxidation. CYP2E1 also plays a role in the metabolism of endogenous compounds including fatty acids and ketone bodies. The regulation of CYP2E1 expression is complex, and involves transcriptional, post-transcriptional, translational, and post-translational mechanisms. CYP2E1 is transcriptionally activated in the first few hours after birth. Xenobiotic inducers elevate CYP2E1 protein levels through both increased translational efficiency and stabilization of the protein from degradation, which appears to occur primarily through ubiquitination and proteasomal degradation. CYP2E1 mRNA and protein levels are altered in response to pathophysiologic conditions by hormones including insulin, glucagon, growth hormone, and leptin, and growth factors including epidermal growth factor and hepatocyte growth factor, providing evidence that CYP2E1 expression is under tight homeostatic control.

• 한국축산식품학회지
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• 제42권2호
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• pp.321-331
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• 2022

Egg yolk is widely used to extract lecithin, which is utilized in the food and cosmetics industry. After lecithin is removed, the rest of egg yolk is generated as a by-product. Thus, it is necessary to properly utilize it. In this study, egg yolk protein extracts were produced using ethanol (EYE-E) and water (EYE-W). Their antioxidant and immunomodulatory effects were then evaluated. Antioxidant activities of EYE-E and EYE-W were determined using cellular antioxidant capacity (CAC) assay and comet assay. EYE-E and EYE-W showed significant (p<0.05) scavenging effects on intracellular reactive oxygen species (ROS) in a dose dependent manner. At a concentration of 50 ㎍/mL, EYE-W showed higher (p<0.05) antioxidant activity than EYE-E. EYE-E and EYE-W also exhibited protective effects against DNA damage caused by oxidative stress. After treatment with EYE-E and EYE-W, DNA damage level of 48.7% due to oxidative stress was decreased to 36.2% and 31.8% levels, respectively. In addition, EYE-E and EYE-W showed immunomodulatory effects by regulating Th1 cytokines (TNF-α and IL-2) and Th2 cytokines (IL-10 and IL-4) in Balb/c mouse splenocytes. These data suggest that EYE-E and EYE-W could be used as functional food ingredients with excellent antioxidant and immunomodulatory activities in the food industry.

• BMB Reports
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• 제32권1호
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• pp.82-85
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• 1999

The movement protein of cucumber mosaic virus (CMV) is required for cell-to-cell movement of viral RNA. The movement of viral RNA occurs through the plant intercellular connection, the plasmodesmata. The viral movement protein was known to be multi-functional. In this work, a series of deletion mutants of CMV movement protein gene were created to identify the functional domains. The mutated movement proteins were produced as inclusion body in E. coli, and purified and renatured. A polyclonal antibody was raised against the CMV-Kor strain (Korean isolate) movement protein expressed in E. coli. The ability of the truncated proteins to bind to ssRNA was assayed by UV cross-linking and gel retardation analyses. The results indicate that the domain between amino acids 118 and 160 of CMV movement protein is essential for ssRNA binding.

• Asian-Australasian Journal of Animal Sciences
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• 제13권4호
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• pp.535-542
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• 2000

Canola meal and seed are poor sources of ruminal undegraded protein (RUP). On average, canola meal and canola seed contains 35 and 14% RUP, respectively. Several protection methods are effective in reducing ruminal degradation of canola protein and in increasing RUP without affecting total tract protein digestibility. Heat (e.g., dry heat, moist heat and jet-sploding) and chemical (e.g., formaldehyde) treatments are the most common methods used to reduce ruminal degradability of canola protein. In most cases, heat treatments were found to be more effective than chemical treatments in protecting canola protein form ruminal degradation. Despite improvement in RUP content and intestinal availability of RUP, data form several studies showed little or no improvement in animal performance as a result of increasing the RUDP level of canola meal and seed.

• Molecules and Cells
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• 제25권3호
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• pp.446-451
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• 2008

We assessed heterologous protein expression in 64 strains obtained from the Escherichia coli Reference (ECOR) collection, a collection representing diverse natural E. coli populations. A plasmid generating a glutathione S-transferase and plant carbonic anhydrase fusion protein (GST-CA) under the control of the tac promoter was introduced into the ECOR strains, and the quantity of the fusion protein was determined by SDS-PAGE. The foreign protein was generated at various levels, from very high (40 strains, high producers) to very low (six strains, low producers). Immunoblotting showed that the high producers expressed approximately 250-500 times more GST-CA protein than the low producers. The results of semi-quantitative RT-PCR showed that the low producers generated mRNA levels comparable to those of the high producers, thereby suggesting that, at least in this case, inefficient translation is a major cause of the low production. We introduced a different plasmid, which expressed a maltose binding protein and plant guanylate kinase fusion protein (MBP-GK) into the six low producers. Interestingly, five of these expressed MBP-GK at very high levels. Thus, we conclude that the production of a particular protein from an expression vector can vary considerably, depending on the host strain. Strains in the ECOR collection could function as useful alternative hosts when a desired level of protein expression is not obtained from commonly used strains, such as E. coli K12 or B derivatives.

• Environmental Analysis Health and Toxicology
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• 제29권
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• pp.10.1-10.6
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• 2014

Objectives Cigarette smoking had been recorded as the main cause of impaired endothelium-dependent vasodilation in smokers by reducing nitric oxide (NO), a production of endothelial nitric oxide synthase (eNOS). However, the mechanism of NO impairment via eNOS activity is unclear until now. In this study, cell passage is suggested to be a relevant factor to eNOS expression under cigarette smoking stress. Methods Bovine aortic endothelial cells (BAECs) were chosen as the research subject with passages ranking from 6 to 9 (6P to 9P). After exposure of cigarette smoking extract (CSE) solution, MTT assay and Western blot method were performed to check the cell viability as well as eNOS protein concentration. In these experiments, four concentrations of CSE at 0.5, 1, 2, and 4% were selected for treatment. Results Our results showed that cells almost died at 4% of CSE. Besides, eNOS protein mass had a linear decrease under the increase of CSE concentration. In addition, the effect of CSE on eNOS expression was dissimilar between different passages. Conclusions This study indicated that CSE had effect on both cell viability and eNOS expression. Besides, a reduction in protein mass was matched with the decrease of cell viability due to CSE tress. Last but not least, the response of eNOS protein to different concentration of CSE at different passages was disparate, making the hypothesis about cell passage related inhibition of eNOS caused by CSE solution.