• IMMUNE NETWORK
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• v.3 no.3
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• pp.219-226
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• 2003

Background: Phage display is the most widely used technique among display methods to produce monoclonal antibody fragment with a specific binding activity. Having a large library for efficient antibody display/selection is quite laborious process to have more than $10^9$ members of transformants. To overcome these limitations, several in vitro selection approaches have been reported. Ribosome display that links phenotypes, proteins, directly to genotype, mRNA, is one of the in vitro display methods. Ribosome display can reach the size of scFv library up to $10^{14}$ molecules and it can be further diversified during PCR steps. To select the high affinity scFv from one pot library, we established ribosome display technique by modifying the previously reported eukaryotic translation system. Methods: To establish the antibody selection system by ribosome display, we used 3D8, anti-DNA antibody. A 3D8 scFv was synthesized in vitro by an in vitro transcription-translation system. The translated 3D8 scFv and the encoding 3D8 mRNA are connected to the ribosome. These scFv-ribosome-mRNA complexes were selected by binding to their specific antigens. The eluted mRNAs from the complexes are reverse transcribed and re-amplified by PCR. To apply this system, antibody library from immunized mouse with terminal protein (TP)-peptide of hepatitis B virus DNA polymerase TP domain was also used. This TP-peptide encompasses the 57~80 amino acid residues of TP. These mRNA/ribosome/scFv complexes by our system were panned three times against TP-peptide. The enrichment of antibody from library was determined by radioimmunoassay. Results: We specifically selected 3D8, anti-DNA antibody, against ssDNA as a model system. The selected 3D8 RNAs sequences from translation complexes were recovered by RT-PCR. By applying this model system, we enriched TP-peptide-specific scFv pools through three cycles of panning from immunized library. Conclusion: We show that our translating ribosome complexes are well maintained and we can enrich the TP-specific scFv pools. This system can be applied to select specific antibody from an antibody library.

• KSBB Journal
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• v.21 no.6 s.101
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• pp.405-409
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• 2006

Bokbunja (Rubus coreanus Miquel) is a wild berry belonging to Rosaceae genus of which application areas have been expanded due to many health effects. Bokbunja berry contains carbohydrates, protein, fats and dietary fibers as major components and various flavonoids such as tannins, volatile components including organic acids, alcohols and hydrocarbons as minor ones. Those active compounds attribute to the following health effects: increase in immune function and antioxidant activity, rise in hormone secretion, suppression of the growth of type B hepatitis virus, and control of weight gain. To utilize the above activities, many products have been developed by using Bokbunja berry extracts in the area of foods, liquors, and cosmetics. Recently, pure compounds have been isolated from the extracts for the development of medicines. In this mini review, the state-of-the-art of Bokbunja researches and applications including papers and patents is summarized.

• Archives of Pharmacal Research
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• v.28 no.4
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• pp.451-457
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• 2005

Experimental studies have demonstrated that the triple combination of amantadine (A)/ ursodeoxycholic acid (UDCA, U)/ biphenyl dimethyl dicarboxylate (DDB, D) might have a preferential antiviral effect compared with that observed in interferon-induced antiviral signal pathways, such as those of $STAT1\alpha$ and the 6-16 genes. To confirm the results, this study examined whether th signal transduction for the antiviral activity in HepG2 2.2.15 was induced dependently or independently of interferon. To accomplish this, the correlation between the $STAT1\alpha$ and 6-16 genes, and nitric oxide, for the mediation of the antiviral activity was assessed. The increase in nitric oxide in the UDCA groups suggests that the inhibition of viral gene replication was enhanced by the amantadine combinations (AU and AUD), and might be more effective if incubated for longer periods. It was found that $STAT1\alpha$ was activated by the amantadine combination, although to a lesser extent than that of $interferon-\alpha$, and the primary endpoints examined for the inhibition of gene expression (HBsAg and HBcAg) were remarkably well regulated. This suggests that the amantadine triple, or at least the double, combination had better clinical benefits than those of $IFN-\alpha$ and the nucleoside analogue single treatment. This demonstrates that the amantadine combination might be a substitute for the existing HBV therapy if the results of in vivo and in vitro studies concur.

• Biomolecules & Therapeutics
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• v.20 no.3
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• pp.273-279
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• 2012

Hesperidin, a flavanone present in citrus fruits, has been studied as potential therapeutic agents that have anti-tumor activity and apoptotic effects in several cancers, but there is no report about the apoptotic effect of hesperidin in human malignant pleural mesothelioma through the specificity protein 1 (Sp1) protein. We investigated whether hesperidin inhibited cell growth and regulated Sp1 target proteins by suppressing the levels of Sp1 protein in MSTO-211H cells. The $IC_{50}$ value of hesperidin was determined to be 152.3 ${\mu}M$ in MSTO-211H cells for 48 h. Our results suggested that hesperidin (0-160 ${\mu}M$) decreased cell viability, and induced apoptotic cell death. Hesperidin increased Sub-$G_1$ population in MSTO-211H cells. Hesperidin significantly suppressed mRNA/protein level of Sp1 and modulated the expression level of the Sp1 regulatory protein such as p27, p21, cyclin D1, Mcl-1, and survivin in mesothelioma cells. Also, hesperidin induced apoptotic signaling including: cleavages of Bid, caspase-3, and PARP, upregulation of Bax, and down-regulation of Bcl-$_{xl}$ in mesothelioma cells. These results show that hesperidin suppressed mesothelioma cell growth through inhibition of Sp1. In this study, we demonstrated that Sp1 acts as a novel molecular target of hesperidin in human malignant pleural mesothelioma.

• Natural Product Sciences
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• v.28 no.4
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• pp.187-193
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• 2022

Lysimachia christinae Hance was commonly used in Oriental medicine for treating the hepatitis virus, cholecystitis and cholagogic efficiency. According to the previous study, it possesses high antioxidant and anti-inflammatory activity. Simultaneous determination analytical method of isolated eight compounds, cynaroside (1), 2-(3,4-dimethoxyphenyl) ethyl O-α-L-arabinopyranosyl-(1→2)-O-[6-deoxy-α-L-mannopyranosyl-(1→3)] β-D-glucopyranoside (2), stearylester ricinoleic acid (3), (E)-4-(3,4-dimethoxyphenyl) but-3-en-1-yl palmitate (4), 2-hydroxy-24-methoxy-4-tetracosenoic acid (5), 2-hydroxy-24-propoxy-4-tetracosenoic acid (6), β-sitosterol (7), and androst-16-ene-3,6-diol (8) were established by using HPLC-DAD. This HPLC analysis was detected on a Dionex C18 column (5 ㎛, 120 Å, 4.6 mm × 150 mm) at 25℃. The mobile phase consisted of 0.1% trifluoroacetic acid and acetonitrile at a flow rate of 1 mL/min. Validation of the method was assessed by linearity, precision and accuracy test. Calibration curve was good at r² > 0.9998. Limits of detection (LOD) ranged from 0.19 to 8.18 g/ml and Limits of quantification (LOQ) ranged from 0.19 to 24.80 g/ml. The relative standard deviations (RSD) values of precision test, intra- and inter- day, were less than 0.99% and 1.0%. The accuracy test results ranged from 98.81% to 106.49% and RSD values were less than 0.95%. These results showed that the HPLC-DAD method was very reliable and accurate for the quantity analysis of eight compounds in L. christinae extract for quality control.

• Korean Journal of Plant Resources
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• v.22 no.5
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• pp.454-460
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• 2009

Phyllanthus urinaria was an important species in Korea and distributed in all around of Korea. The roots and stems of this plant have been used for natural medicine for the treatment of diabetes, the hepatitis B virus and disturbances of the kidney and urinary bladder. Production of adventitious roots in P. urinaria by in vitro cultures could be used as alternatives materials. Shoot and root segments from P. urinaria seedling were cultured on Murashige and Skoog (MS) medium supplemented with 3.0 mg/L IBA and 30 g/L sucrose. After 4 weeks of culture, the highest induction of adventitious roots was obtained from the shoot part. Frequency of adventitious root formation on medium with various kinds of auxins (IAA, NAA, 2,4-D, and IBA) and various concentrations of IBA (0, 0.1, 0.5, 1.0, 3.0, and 5.0 mg/L) was tested. The maximun induction of adventitious root was obtained on medium with 0.5 mg/L IBA. In liquid culture, growth of root was best on medium supplemented with 30 g/L sucrose. Adventitious roots were cultured in 5 L bioreactor containing 1/2 MS medium supplemented with 0.5 mg/L IBA and 30 g/L sucrose and mass-production of adventitious roots was successfully achieved. These results revealed the first attempt for the production of adventitious roots in P. urinaria.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.11
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• pp.1651-1659
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• 2017

Objective: Lao PDR's recent accession to the World Trade Organization necessitates a greater understanding of the patterns and risk of livestock production in order to better align national policy with the Agreement on the Application of Sanitary and Phytosanitary Measures. This eco-health study was conducted to improve understanding of the interrelations between market chains and zoonotic infection risks at two strategic cross border points between Lao PDR, Thailand and Viet Nam. Methods: Information gained from smallholder farmer/trader interviews was integrated with serological surveys for pig-associated zoonoses-including hepatitis E virus (HEV), Taenia solium (T. solium) and trichinella-to identify potential linkages between disease risk and pig production and slaughter in low input systems common across the country. Results: Trichinella and HEV exposure was high in both humans and pigs in both study areas, significantly associated with pig slaughter and the subsequent consumption and handling of raw pork products. T. solium demonstrated a strong geographical and ethnic association with the northern study area bordering Vietnam. With the right knowledge and accessible, affordable inputs, the majority of smallholder farmers indicated a willingness to invest more in pig production, which could simultaneously improve livelihoods and decrease exposure to HEV, Trichinella, and T. solium through increased access to formal markets and an improved slaughter processes. Conclusion: The linkages identified when assessing disease risk in the context of potential economic and cultural drivers of transmission highlight the importance of a systems-based approach for the detection and control of zoonotic disease, and contributes to an improved understanding of the Lao PDR livestock sector.