• Food Science and Preservation
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• v.15 no.6
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• pp.891-896
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• 2008

Total phenolics and flavonoids, and the antioxidant capacity of plum cultivar wines (Prunus salicina L. cv. Soldam and P. salicina L. cv. Formosa) were determined using spectrophotometric methods. The total phenolic and flavanoid contents of Soldam wine were $478.4\;{\pm}\;5.6\;mg$ GAE and $202.4\;{\pm}\;7.5\;mg$ CE per L,respectively, and in Formosa wine were $200.6\;{\pm}\;7.5\;mg$ GAE and $64.4\;{\pm}\;6.8\;mg$ CE per L, respectively. Neutral and acidic phenolics in Soldam wine were extracted with ethyl acetate and 0.01 N HCl, respectively. In the 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging assay, neutral phenolics (64.5 EDA%) had $3{\sim}4$ times higher antioxidant activity than acidic phenolics (21.5 EDA%) and other related phenolic compounds such as chlorogenic acid (15.5 EDA%) and quercetin (24.6 EDA%) at a concentration of $100\;{\mu}g/mL$. The antiviral activities of neutral and acidic phenolics in Soldam wine were investigated in vitro using a virus-induced cytopathic effect (CPE) inhibition assay. Results showed that neutral and acidic phenolics at concentrations of $100\;{\mu}g/mL$ inhibited porcine epidemic diarrhea virus (PEDV) replication at rates of 78.12% and 58.37%, respectively. The inhibition rate of 10 g/mL neutral phenolics (69.42%) was higher than that of ribavirin as an antiviral reagent (57.86%). At concentrations of $100\;{\mu}g/mL$ or less, neutral and acidic phenolics of Soldam wine had no cytotoxic effect against vero cells.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.153-163
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• 2002

In vitro matured porcine oocytes have very small volume of perivitellinspace (PVS). In these respect, the effect of sucrose and polybrene on the efficiency of gene transfer was investigated. As a gene (hGH) transfer vehicle, vesicular stomatitis virus glycoprotein pseudotyped retroviral vector (VSV-G) was used. Sucrose treatment has no detrimental effect on the rates of cleavage and resulted in the enlargement of PVS for the efficient introduction of retroviral vector stocks. Introduction rates of retrovirus in 0.5, 1, 2, 3 % sucrose treatment group were higher than that of the non-treatment group (39.3, 43.3, 35.7, 40.7 % vs. 8.3 %), respectively. In addition, we observed that sucrose pretreatment during injection procedure significantly reduce the frequency of polyspermy. In general, polybrene is a polycation essential for retrovirus transduction. The groups with the addition of 0.5, 5, 50$\mu\textrm{g}$/$m\ell$ polybrene exhibited a significant effect on gene transfer compared to that of the non-addition group (56.5, 50.0, 57.1 % vs. 34.6 %), respectively But, when the oocytes were co-injected with retrovirus and 50$\mu\textrm{g}$/$m\ell$ polybrene, the rates of cleavage and blastocyst development were 43.3 and 4.6%, respectively. This rates were lower than those of the non-addition group (70.0 and 17.3 %). In conclusion, sucrose pretreatment have increased efficiency of retroviral mediated gene transfer in porcine oocytes with no damage on in vitro fertilization and embryo development. In addition, sucrose pretreatment was beneficial in polyspermy inhibition. Presence of polybrene during microinjection showed a beneficial effect on the gene transfer in porcine oocytes, in low concentration. And these results will provide an useful tool for production of transgenic pigs by retroviral mediated gene transfer.

• Korean Chemical Engineering Research
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• v.50 no.5
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• pp.866-873
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• 2012

IgY (Immunoglobulin Yolk) is a specific antibody in egg yolk, and it protects human body from virus and antigen. There are a lot of egg yolk components such as lipoprotein and protein. To separate IgY, HPLC (High Performance Liquid Chromatography) and precipitation were used in a batch mode and SMB (Simulated Moving Bed) was adopted for continuous purification of yolk proteins. IgY and other proteins in yolk were separated by using three-zone and four-zone SMB chromatography. Before performing SMB experiments, batch chromatography simulation parameters and adsorption isotherms were obtained. The parameters of batch chromatography were used to simulate SMB using Aspen chromatography. To compare three-zone and four-zone SMB chromatography, simulations in $m_2-m_3$ plane on the triangle theory were carried out. In terms of concentration and purity of both IgY and other lipoproteins, 3-zone SMB process is considered as ideal at the vertex of triangle ($m_2$, $m_3$=0.1, 1.1). 4-zone SMB yields the highest IgY purity at the coordinate ($m_2$, $m_3$=0.06, 0.5), which is the pure raffinate region. In 3-zone SMB without recycle, other lipoproteins in extract are largely affected in purity by small shift from the vertex of triangle ($m_2$, $m_3$=0.1, 1.1).

• Korean Journal of Veterinary Research
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• v.40 no.3
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• pp.471-478
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• 2000

The pharmacokinetic properties of KR-V compounds, recently developed as new anti-HIV agents, were studied after i.v. and p.o. administration in rats. The concentrations of the KR-V series were determined in rat plasma using an high-performance liquid chromatography (HPLC)-UV detection system. Of the 19 KR-V compounds investigated in the present study, only KR-V 3, 10, 14, 16 and 18-1 showed oral bioavailability. The plasma concentration-time data could be adequately described by an one-compartment open model. In the i.v. kinetic study (10mg/kg), the CLt of KR-V 3, 10, 14 and 16 (>4L/hr/kg) were significantly higher than that of KR-V 18-1 (1.1 L/hr/kg). The AUC of KR-V 18-1 was greater ($8.97{\mu}g{\cdot}hr/ml$) than that of the other compounds, but the Vd (0.58 L/kg) was lower. In the p.o. kinetic study (50mg/kg), although the t-1/2 of KR-V 18-1 was shorter than that of the other compounds, the AUC ($3.659{\mu}g{\cdot}hr/ml)$ and $C_{max}(1.891{\mu}g/ml$) were markedly higher. In a seperated in vitro experiment, only KR-V 18-1, of the 5 compounds with bioavailibility, exhibits potent activity against HIV-1 mutant strains. Therefore, KR-V 18-1 is expected to become a new potent anti-AIDS drug candidate/lead compound.

• Korean Journal of Food Science and Technology
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• v.39 no.1
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• pp.71-76
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• 2007

Foodborne illness caused by Noroviruses (NVs) is increasing rapidly in Korea. This study developed an effective detection protocol for NVs found in contaminated oysters and lettuce through an investigation using the major steps of virus particle separation, concentration and RT-PCR. As a surrogate model for NVs, the cultivable feline calicivirus (FCV) that belongs to the same Caliciviridae family was used. Instead of using a time-consuming ultracentrifugation method, efficient methods based on solvent extraction and PEG precipitation procedure were applied. Direct homogenization of a 25g sample of whole oyster and lettuce in 175mL PBS provided the simplicity that would be needed in the actual field of food product examination. The overnight PEG precipitation step at $4^{\circ}C$ was reduced to 3 h by placing the reaction tube in ice and by adjusting the PEG concentrations. The application of the use of chloroform and 0.2 ${\mu}m$ syringe filtration together showed a better detection efficiency than the use of chloroform alone in removing PCR inhibitors for both oyster and lettuce samples. Also, dilution of the extracted RNA solution before PCR provided increased sensitivity. The improved detection protocol developed in this study could be efficiently applied to detect FCV and most likely NVs from oysters and lettuce.

• Journal of Plant Biotechnology
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• v.35 no.1
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• pp.75-80
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• 2008

In vitro culture of shoot tip of garlic (Allium sativium L. cv. Seosan) was carried out to find medium condition of the induction of multiple shoots and bulbing for muliproduction of virus-free seed bulbs. For this work, tank culture was introduced. In shoot tip culture on MS solid medium the induction of multiple shoots and bulbing were better by adding 3% sucrose than 8%. Supplementation with 2mg/L 2ip and 0.2 mg/L IAA in this medium was effective. Three point three shoots including 2.7 bulbs were formed from a shoot tip after cultivation for 30 days on this medium. Bulbing of garlic in liquid culture with plastic water tank of 20L supplied air at the side of the lower part was better by adding 3% sucrose than 8% by subculture for 45 days with shoots obtained from shoot tip culture for 30 days on soid MS medium. Shoot growth was vigorous at 3% sucrose however bulb growth was more effective on the medium of 8% sucrose. Because of the effectiveness on solid medium added 3% sucrose, 2 mg/L 2ip and 0.2 mg/L IAA for initial production of multi-shoot in stem tip culture and the effectiveness in liquid culture with water tank for growth of bulbs, the method of two-step culture could be introduced for the multiple production of seed bulb of high quality. It was more desirable by supply of 0.2 mg/L BA and 0.02 mg/L NAA at tank culture time. But growth of the bulbs became poor by increasing concentration of NAA of the medium.

• Journal of Life Science
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• v.28 no.4
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• pp.478-482
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• 2018

Human rotavirus is a causative agent of acute diarrhea among children. The artificial gene encoding the truncated $VP8^*$ protein of human rotavirus A (serotype 1 strain WA) was synthesized according to the Escherichia coli codon preference. The synthetic $VP8^*$ gene also possessed the NdeI and HindIII restriction sites for the convenient in-frame cloning for translation and a 6-histidine tag at C-terminus for Ni+ affinity purification. Molecular weight of the truncated $VP8^*$ protein deduced from the nucleotide sequences of the artificial gene was a 19.7-kDa. This synthetic $VP8^*$ DNA fragment was inserted into the pT7-7 expression vector and transformed into E. coli BL21 (DE3). Transformants harboring the synthetic gene encoding the $VP8^*$ protein was induced by supplement of a final concentration of 0.05 mM ITPG at $20^{\circ}C$. Protein crude extract from the E. coli transformants was subjected to Western blotting with the mouse anti-rotavirus capsid antibody, showing ~20-kDa $VP8^*$ protein band. The truncated $VP8^*$ protein band was also observed by Western blotting using the rabbit polyclonal antibody serum made against the truncated $VP8^*$ protein. This study suggested that the synthetic gene could be used as an easy way to produce the antigenic vaccine candidate for control of virus-associated diseases or to develop antibodies for diagnostic purpose.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2002.07a
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• pp.113-113
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• 2002

Phylogenetically conserved Bcl-2 family proteins play a pivotal role in the regulation of apoptosis from virus to human. Members of the Bcl-2 family consist of antiapoptotic proteins such as Bcl-2, Bcl-xL, and Bcl-w, and proapoptotic proteins such as BAD, Bax, BOD, and Bok. It has been proposed that anti- and proapoptotic Bcl-2 proteins regulate cell death by binding to each other and forming heterodimers. A delicate balance between anti- and proapoptotic Bcl-2 family members exists in each cell and the relative concentration of these two groups of proteins determines whether the cell survives or undergoes apoptosis. Mcl-1 (Myeloid cell :leukemia-1) is a member of the Bcl-2 family proteins and was originally cloned as a differentiation-induced early gene that was activated in the human myeloblastic leukemia cell line, ML-1 . Mcl-1 is expressed in a wide variety of tissues and cells including neoplastic ones. We recently identified a short splicing variant of Mcl-1 short (Mcl-IS) and designated the known Mcl-1 as Mcl-1 long (Mcl-lL). Mcl-lL protein exhibits antiapoptotic activity and possesses the BH (Bcl-2 homology) 1, BH2, BH3, and transmembrane (TM) domains found in related Bcl-2 proteins. In contrast, Mcl-1 S is a BH3 domain-only proapoptotic protein that heterodimerizes with Mcl-lL. Although both Mc1-lL and Mcl-lS proteins contain BH domains fecund in other Bcl-2 family proteins, they are distinguished by their unusually long N-terminal sequences containing PEST (proline, glutamic acid, serine, and threonine) motifs, four pairs of arginine residues, and alanine- and glycine-rich regions. In addition, the expression pattern of Mcl-1 protein is different from that of Bcl-2 suggesting a unique role (or Mcl-1 in apoptosis regulation. Tankyrasel (TRF1-interacting, ankyrin-related ADP-related polymerasel) was originally isolated based on its binding to TRF 1 (telomeric repeat binding factor-1) and contains the sterile alpha motif (SAM) module, 24 ankyrin (ANK) repeats, and the catalytic domain of poly(adenosine diphosphate-ribose) polymerase (PARP). Previous studies showed that tankyrasel promotes telomere elongation in human cells presumably by inhibiting TRFI though its poly(ADP-ribosyl)action by tankyrasel . In addition, tankyrasel poly(ADP-ribosyl)ates Insulin-responsive amino peptidase (IRAP), a resident protein of GLUT4 vesicles, and insulin stimulates the PARP activity of tankyrase1 through its phosphorylation by mitogen-activated protein kinase (MAPK). ADP-ribosylation is a posttranslational modification that usually results in a loss of protein activity presumably by enhancing protein turnover. However, little information is available regarding the physiological function(s) of tankyrase1 other than as a PARP enzyme. In the present study, we found tankyrasel as a specific-binding protein of Mcl-1 Overexpression of tankyrasel led to the inhibition of both the apoptotic activity of Mel-lS and the survival action of Mcl-lL in mammalian cells. Unlike other known tankyrasel-interacting proteins, tankyrasel did not poly(ADP-ribosyl)ate either of the Mcl-1 proteins despite its ability to decrease Mcl-1 proteins expression following coexpression. Therefore, this study provides a novel mechanism to regulate Mcl-1-modulated apoptosis in which tankyrasel downregulates the expression of Mcl-1 proteins without the involvement of its ADP-ribosylation activity.

• Journal of Food Hygiene and Safety
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• v.30 no.2
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• pp.150-158
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• 2015

This article reports the development of an effective test procedure for detection of norovirus (NoV) in foods of diverse matrices. In this study, target foods included fermented milk, soybean paste, powders made from uncooked grains and vegetables, sesame leaves preserved in soy sauce, pickled mooli, and mooli. Viral recovery varied depending on the food matrices or elution buffers tested. Buffers were compared to determine effective elution buffers from artificially virus-contaminated foods. The conventional test procedure for concentrating viruses from food (elution-polyethylene glycol(PEG) precipitation-chloroform-PEG precipitation) was modified to save time by eliminating one PEG precipitation step. The modified procedure (elution-chloroform-PEG precipitation) was able to concentrate viruses more effectively than the conventional procedure. It also removed RT-PCR inhibitors effectively. The modified procedure was applied to target food for genogroup II NoV detection. NoV RNA was detected at the initial inoculum levels 3.125-12.5 RT-PCR units per 10-25 g tested food. The use of this newly established procedure should facilitate detection of low levels of norovirus in diverse foods.

• Korean Journal of Food Science and Technology
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• v.37 no.6
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• pp.1018-1023
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• 2005

Simplified procedure was developed for concentrating and detecting poliovirus and norovirus in oysters. Viruses were seeded into oyster tissue homogenates and concentrated through polyethylene glycol (PEG) precipitation, chloroform or Freon extraction, with additional PEG precipitation. Amount of viruses was evaluated using poliovirus plaque assay. Virus recovery during concentration procedure was approximately 16.4-26.0%. For defection, viral RNAs in oysters were examined using one-step RT-PCR after extraction with Trizol. Dilution or capturing of viral RNA using silica gel membrane allowed viruses to be detected by RT-PCR, whereas viruses could not be removed using $QIAshredder^{TM}$ Homogenizer, which is effective in removing RT-PCR inhibitors in lettuce and hamburgers. Freon extraction, generally used to concentrate viruses found in food, could be substituted with chloroform extraction using this procedure; no difference could be observed between detection limits of whole oyster extracts and digestive organ extracts indicating that RT-PCR inhibitors were distributed evenly throughout whole tissues. Nested PCR greatly improved efficiency of this procedure. Overall, this procedure could remove sufficient amount of inhibitors to allow detection of norovirus in oysters.