Bioconversion of ginsenoside Rc into Rd by a novel α-L-arabinofuranosidase, Abf22-3 from Leuconostoc sp. 22-3: cloning, expression, and enzyme characterization.

Liu, Qing-Mei,Jung, Hae-Min,Cui, Chang-Hao,Sung, Bong-Hyun,Kim, Jin-Kwang,Kim, Song-Gun,Lee, Sung-Taik,Kim, Sun-Chang,Im, Wan-Taek N.V. Swets en Zeitlinger 2013 Antonie van Leeuwenhoek Vol.103 No.4

<P>A novel α-L-arabinofuranosidase (Abf22-3) that could biotransform ginsenoside Rc into Rd was obtained from the ginsenoside converting Leuconostoc sp. strain 22-3, isolated from the Korean fermented food kimchi. The gene, termed abf22-3, consisting of 1,527 bp and encoding a protein with a predicted molecular mass of 58,486 Da was cloned into the pMAL-c2x (TEV) vector. A BLAST search using the Abf22-3's amino acid sequence revealed significant homology to that of family 51 glycoside hydrolases. The over-expressed recombinant Abf22-3 in Escherichia coli BL21 (DE3) catalyzed the hydrolysis of the arabinofuranoside moiety attached to the C-20 position of ginsenoside Rc under optimal conditions of pH 6.0 and 30 C. This result indicated that Abf22-3 selectively converts ginsenoside Rc into Rd, but did not catalyze the hydrolysis of glucopyranosyl groups from Rc or other ginsenosides such as Rb1 and Rb2. Over-expressed recombinant enzymes were purified by two steps with amylose-affinity and DEAE-cellulose chromatography and then characterized. The kinetic parameters for α-L-arabinofuranosidase showed apparent Km and Vmax values of 0.95 0.02 μM and 1.2 0.1 μmol min(-1) mg of protein(-1) against p-nitrophenyl-α-L-arabinofuranoside, respectively. Using a purified MBP-Abf22-3 (10 μg/ml), 0.1 % of ginsenoside Rc was completely converted to ginsenoside Rd within 20 min.</P>

Novel enzymatic elimination method for the chromatographic purification of ginsenoside Rb₃ in an isomeric mixture

Cui, Chang-Hao,Fu, Yaoyao,Jeon, Byeong-Min,Kim, Sun-Chang,Im, Wan-Taek The Korean Society of Ginseng 2020 Journal of Ginseng Research Vol.44 No.6

Background: The separation of isomeric compounds from a mixture is a recurring problem in chemistry and phytochemistry research. The purification of pharmacologically active ginsenoside Rb₃ from ginseng extracts is limited by the co-existence of its isomer Rb₂. The aim of the present study was to develop an enzymatic elimination-combined purification method to obtain pure Rb₃ from a mixture of isomers. Methods: To isolate Rb₃ from the isomeric mixture, a simple enzymatic selective elimination method was used. A ginsenoside-transforming glycoside hydrolase (Bgp2) was employed to selectively hydrolyze Rb₂ into ginsenoside Rd. Ginsenoside Rb₃ was then efficiently separated from the mixture using a traditional chromatographic method. Results: Chromatographic purification of Rb₃ was achieved using this novel enzymatic elimination-combined method, with 58.6-times higher yield and 13.1% less time than those of the traditional chromatographic method, with a lower minimum column length for purification. The novelty of this study was the use of a recombinant glycosidase for the selective elimination of the isomer. The isolated ginsenoside Rb₃ can be used in further pharmaceutical studies. Conclusions: Herein, we demonstrated a novel enzymatic elimination-combined purification method for the chromatographic purification of ginsenoside Rb₃. This method can also be applied to purify other isomeric glycoconjugates in mixtures.

Inhibition of Transient Receptor Potential Melastain 7 Enhances Apoptosis Induced by TRAIL in PC-3 cells

Lin, Chang-Ming,Ma, Ji-Min,Zhang, Li,Hao, Zong-Yao,Zhou, Jun,Zhou, Zhen-Yu,Shi, Hao-Qiang,Zhang, Yi-Fei,Shao, En-Ming,Liang, Chao-Zhao Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.10

Transient receptor potential melastain 7 (TRPM7) is a bifunctional protein with dual structure of both ion channel and protein kinase, participating in a wide variety of diseases including cancer. Recent researches have reported the mechanism of TRPM7 in human cancers. However, the correlation between TRPM7 and prostate cancer (PCa) has not been well studied. The objective of this study was to investigate the potential the role of TRPM7 in the apoptosis of PC-3 cells, which is the key cell of advanced metastatic PCa. In this study, we demonstrated the influence and potential function of TRPM7 on the PC-3 cells apoptosis induced by TNF-related apoptosis inducing-ligand (TRAIL). The study also found a novel up-regulated expression of TRPM7 in PC-3 cells after treating with TRAIL. Suppression of TRPM7 by TRPM7 non-specific inhibitors ($Gd^{3+}$ or 2-aminoethoxy diphenylborate (2-APB) ) not only markedly eliminated TRPM7 expression level, but also increased the apoptosis of TRAIL-treated PC-3 cells, which may be regulated by the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway accompany with up-regulated expression of cleaved Caspase-3, (TRAIL-receptor 1, death receptors 4) DR4, and (TRAIL-receptor 2, death receptors 5) DR5. Taken together, our findings strongly suggested that TRPM7 was involved in the apoptosis of PC-3 cells induced by TRAIL, indicating that TRPM7 may be applied as a therapeutic target for PCa.

Novel Heterogeneous Recommender using Deep Representations for Music Streaming Services

Yuan-Hao Chan,Jia-Wei Chang,Hung-Min Sun,Chiu, Ching-Yi 한국정보기술학회 2019 Proceedings of The International Workshop on Futur Vol.2018 No.1

Association Between TP53 Arg72Pro Polymorphism and Hepatocellular Carcinoma Risk: A Meta-analysis

Xu, Chang-Tao,Zheng, Fang,Dai, Xin,Du, Ji-Dong,Liu, Hao-Run,Zhao, Li,Li, Wei-Min Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.9

Background: Previous studies on the association between the TP53 Arg72Pro polymorphism and hepatocellular carcinoma (HCC) risk obtained controversial findings. This study aimed to quantify the strength of the association by meta-analysis. Methods: We searched PubMed and Wangfang databases for published studies on the association between the TP53 Arg72Pro polymorphism and HCC risk, using the pooled odds ratio (OR) with its 95% confidence intervals (95% CI) for assessment. Results: 10 studies with a total of 2,026 cases and 2,733 controls were finally included into this meta-analysis. Overall, the TP53 Arg72Pro polymorphism was not associated with HCC risk (all P values greaterth HCC risk in Caucasians in three genetic models (For Pro versus Arg, OR = 1.20, 95%CI 1.03-1.41; For ProPro versus ArgArg, OR = 1.74, 95%CI 1.23-2.47; For ProPro versus ArgPro/ArgArg, OR = 1.85, 95%CI 1.33-2.57). However, there was no significant association between the TP53 Arg72Pro polymorphism and HCC risk in East Asians (all P values greater than 0.10). No evidence of publication bias was observed. Conclusion: Meta-analyses of available data suggest an obvious association between the TP53 Arg72Pro and HCC risk in Caucasians. However, the TP53 Arg72Pro polymorphism may have a race-specific effect on HCC risk and further studies are needed to elucidate this possible effect.

An Efficient Implementation of Mobile Raspberry Pi Hadoop Clusters for Robust and Augmented Computing Performance

Srinivasan, Kathiravan,Chang, Chuan-Yu,Huang, Chao-Hsi,Chang, Min-Hao,Sharma, Anant,Ankur, Avinash Korea Information Processing Society 2018 Journal of information processing systems Vol.14 No.4

Rapid advances in science and technology with exponential development of smart mobile devices, workstations, supercomputers, smart gadgets and network servers has been witnessed over the past few years. The sudden increase in the Internet population and manifold growth in internet speeds has occasioned the generation of an enormous amount of data, now termed 'big data'. Given this scenario, storage of data on local servers or a personal computer is an issue, which can be resolved by utilizing cloud computing. At present, there are several cloud computing service providers available to resolve the big data issues. This paper establishes a framework that builds Hadoop clusters on the new single-board computer (SBC) Mobile Raspberry Pi. Moreover, these clusters offer facilities for storage as well as computing. Besides the fact that the regular data centers require large amounts of energy for operation, they also need cooling equipment and occupy prime real estate. However, this energy consumption scenario and the physical space constraints can be solved by employing a Mobile Raspberry Pi with Hadoop clusters that provides a cost-effective, low-power, high-speed solution along with micro-data center support for big data. Hadoop provides the required modules for the distributed processing of big data by deploying map-reduce programming approaches. In this work, the performance of SBC clusters and a single computer were compared. It can be observed from the experimental data that the SBC clusters exemplify superior performance to a single computer, by around 20%. Furthermore, the cluster processing speed for large volumes of data can be enhanced by escalating the number of SBC nodes. Data storage is accomplished by using a Hadoop Distributed File System (HDFS), which offers more flexibility and greater scalability than a single computer system.

Reduction of Physical Strength and Enhancement of Anti-Protein and Anti-Lipid Adsorption Abilities of Contact Lenses by Adding 2-Methacryloyloxyethyl Phosphorylcholine

Wan-Hsin Chang,Pei-Yi Liu,Chien-Ju Lu,Dai-En Lin,Min-Hsuan Lin,Yuan-Ting Jiang,Yuan-Hao Howard Hsu 한국고분자학회 2020 Macromolecular Research Vol.28 No.12

Biocompatible 2-methacryloyloxyethyl phosphorylcholine (MPC) can enhance the adsorption of water molecules and is therefore used for manufacturing contact lenses. This study investigated the mechanical strength, anti-protein deposition, and anti-lipid adsorption effects of MPC addition to contact lenses. Experimental contact lenses produced by copolymerizing multiple ratios of MPC to 2-hydroxyethyl methacrylate (HEMA) were analyzed. Atomic force microscopy revealed that MPC addition increased surface roughness. The anti-protein deposition and anti-lipid adsorption effects on poly(HEMA-MPC) polymers of various phosphorylcholine quantities were experimentally confirmed. The water content of the contact lenses was proportional to the MPC content in the polymer. The hydrated PC moiety of MPC drastically altered the network of the poly-HEMA polymer by inserting water molecules, which were trapped in the concave region of the surface. MPC addition had negative effects on all examined strength factors because of structural destabilization of the copolymer through water insertion. The anti-deposition effects of MPC were verified by examining the lysozyme and lipid adsorption abilities of the prepared contact lenses. Our results revealed that MPC enhanced interactions of the poly(HEMA-MPC) copolymer with water molecules; these interactions weakened the mechanical strength of the copolymer but markedly improved the anti-adsorption property of the biomolecules. The optimal proportion of HEMA–MPC for contact lenses is in the range 14.9%-28.5%.

High Frequency Plant Regeneration from Leaf, Petiole and Internode Explants of Codonopsis lanceolata Benth.

Bimal Kumar Ghimire,Chul-Min Shin,Cheng Hao Li,Na-Young Kim,III-Min Chung,Jung-Dae Lim,Jae-Kwang Kim,Myong-Jo Kim,Dong-Ha Cho,Chang-Yeon Yu 한국약용작물학회 2007 한국약용작물학회지 Vol.15 No.2

An efficient regeneration system was developed using leaf, petiole, and internode explants. Highly embryogenic callus was obtained following cultivation on MS basal nutrient supplemented with 2 mg/l 2,4-D. Globular, heart, torpedo and cotyledon shaped somatic embryo were produced from the surface of embryogenic callus. Direct shoot regeneration without intermediate callus formation has been achieved on MS medium supplemented NAA and BAP. The percentage of response varies with different concentration of auxin and cytokinin treated individually or in combination. The best shoot regeneration response (54.28%) and number of shoot per explant (12.67) were achieved on the medium supplemented with 0.1 mg/l NAA and 1 mg/l BAP. The regenerated shoot transformed into young plant when cultured into elongation and root induction medium. More than 90% of in vitro propagated plants could survive when transferred to the greenhouse for acclimation. This optimized regeneration system can be used for rapid shoot proliferation and genetic transformation.

Malignant transformation of ovarian mature cystic teratoma into squamous cell carcinoma: a Taiwanese Gynecologic Oncology Group (TGOG) study

An Jen Chiang,Min-Yu Chen,Chia-Sui Weng,Hao Lin,Chien-Hsing Lu,Peng-Hui Wang,Yu-Fang Huang,Ying-Cheng Chiang,Mu-Hsien Yu,Chih-Long Chang 대한부인종양학회 2017 Journal of Gynecologic Oncology Vol.28 No.5

Objective: The malignant transformation (MT) of ovarian mature cystic teratoma (MCT)to squamous cell carcinoma (SCC) is very rare. This study analyzed cases from multiplemedical centers in Taiwan to investigate the clinicopathologic characteristics, treatment, andprognostic factors of this disease and reviewed related literature. Methods: Pathological reports of 16,001 patients with primary ovarian cancer who weretreated at Taiwan medical centers from 1990 to 2011 were reviewed. In total, 52 patients withMT of MCT to SCC were identified. Results: Among all ovarian MCTs, the incidence of MT to SCC is 0.2%. The median age ofpatients was 52 years (range, 29–89 years), and the mean tumor size was 10.5 cm (range, 1–40cm). We analyzed the patients in our study and those in the literature and determined thatearly identification and complete surgical resection of the tumor are essential for long-termsurvival. In addition, adjuvant chemotherapy or concurrent chemoradiotherapy can be usedto treat this malignancy. Old age, large tumor size (≥15.0 cm), and solid components in MCTsare suitable indicators predicting the risk of MT of MCT to SCC. Conclusion: Similar to general epithelial ovarian cancers, the early detection of MT of MCTto SCC is critical to long-term survival. Therefore, older patients with a large tumor or those with a tumor containing a solid component in a clinically diagnosed MCT should beevaluated to exclude potential MT to SCC.

Field Performance and Morphological Characterization of Transgenic Codonopsis lanceolata Expressing γ-TMT Gene.

Bimal Kumar Ghimire,Cheng Hao Li,Hyun-Young Kil,Na-Young Kim,Jung-Dae Lim,Jae-Kwang Kim,Myong-Jo Kim,Ill-Min Chung,Sun-Joo Lee,Seok-Hyun Eom,Dong-Ha Cho,Chang-Yeon Yu 한국약용작물학회 2007 한국약용작물학회지 Vol.15 No.5

Field performance and morphological characterization was conducted on seven transgenic lines of Codonopsis lanceolata expressing γ-TMT gene. The shoots were obtained from leaf explants after co-cultivation with Agrobacterium tume-faciens strain LBA 4404 harboring a binary vector pYBI 121 that carried genes encoding γ-Tocopherol methyltransferase gene (γ-TMT) and a neomycin phosphotransferase II gene (npt II) for kanamycin resistance. The transgenic plants were transferred to a green house for acclimation. Integration of T-DNA into the T0 and T1 generation of transgenic Codonopsis lanceolata genome was confirmed by the polymerase chain reaction and southern blot analysis. The progenies of transgenic plants showed phenotypic differences within the different lines and with relative to control plants. When grown in field, the transgenic plants in general exhibited increased fertility, significant improvement in the shoot weight, root weight, shoot height and rachis length with relation to the control plants. However, all seven independently derived transgenic lines produced normal flower with respect to its shape, size, color and seeds number at its maturity. Indicating that the addition of a selectable marker gene in the plant genome does not effect on seed germination and agronomic performance of transgenic Codonopsis lanceolata. T1 progenies of these plants were obtained and evaluated together with control plant in a field experiment. Overall, the agronomic performance of T1 progenies of transgenic Codonopsis lanceolata showed superior to that of the seed derived non-transgenic plant. In this study, we report on the morphological variation and agronomic performance of transgenic Codonopsis lanceolata developed by Agrobacterium transformation.