Broad-specificity amino acid racemase, a novel non-antibiotic selectable marker for transgenic plants

Yi‑Chia Kuan,Venkatesan Thiruvengadam,Jia‑Shin Lin,Jia‑Hsin Liu,Tsan‑Jan Chen,Hsin‑Mao Wu,Wen‑Ching Wang,Liang‑Jwu Chen 한국식물생명공학회 2018 Plant biotechnology reports Vol.12 No.1

The broad-specificity amino acid racemase (Bsar) from Pseudomonas putida catalyzes the racemization of various amino acids, offering a flexible and feasible platform to develop a new non-antibiotic selectable marker system for plant transformation. In the present study, we demonstrated that a Bsar variant, Bsar-R174K, that is useful as a selectable marker gene in Arabidopsis and rice that were susceptible to l-lysine and D-alanine. The introduction of wild-type Bsar, Bsar-R174K or Bsar-R174A into E. coli lysine or asparagine auxotrophs was able to rescue the growth of these microorganisms in minimal media supplemented with selectable amino acid enantiomers. The transformation of Arabidopsis with Bsar or Bsar variants based on d-alanine selection revealed that Bsar-R174K had the greatest efficiency (2.40%), superior to kanamycin selectionbased transformation (1.10%). Whereas, l-lysine-based selection exhibited lower efficiency for Bsar-R174K (0.17%). The progenies of selected Bsar-R174K transgenic Arabidopsis revealed normal growth properties. In addition, Bsar-R174K transgenic rice was obtained on l-lysine medium with an efficiency of 0.9%, and the progenies of the transgenic rice revealed morphologically normal phenotypes comparable with their wild-type counterparts. This study presents the first report of broad range amino acid racemase Bsar-R174K as a non-antibiotic selectable marker system applied in transgenic plants.

Comparison of stomach microRNA transcriptomes of Tibetan and Yorkshire pigs by deep sequencing

Wen‑kui Sun,Yanyue Li,Chi Cheng,Yi‑hui Chen,Kai Zeng,Xiaohui Chen,Yiren Gu,Rui Liu,Xuebin Lv,Rong Gao 한국유전학회 2018 Genes & Genomics Vol.40 No.9

MiRNAs regulate the expression of target genes in diverse cellular processes and hence play important roles in different physiological processes, yet little is known about the stomach microRNAome (miRNAome) of the Tibetan pig. The objective of this experiment was to investigate differentially expressed stomach miRNAs participating in digestion. Firstly, we isolated total RNA by Trizol reagent from three Tibetan and three Yorkshire purebred pigs stomach samples at 90-day-old. Secondly, a comprehensive analysis of Tibetan and Yorkshire pig stomach miRNAomes was performed by small RNA sequencing in the Illumina HiSeq 2000 system. Finally, SYBR Green Real-time RT-PCR was performed to validate the differentially expressed miRNAs. We identified 318 unique miRNAs, 260 were co-expressed in both libraries, 17 and 31 miRNAs were specifically expressed in Tibetan and Yorkshire pigs respectively. Fifty six differentially expressed miRNAs were identified by the identifying differentially expressed genes 6 (IDEG6). Kyoto encyclopedia of genes and genomes analysis revealed that some of the differentially expressed miRNAs were associated with protein and fat digestion. Two differentially expressed miRNAs (miR-214-3p and ssc-un39) participating in the digestion of lipid were identified. Additionally, qRT-PCR results suggested that a higher expression of miR-214-3p in the Tibetan pig stomach could lead to relatively lower expression of calcium-dependent phospholipase A2, which is an enzyme important for the digestion of glycerol phospholipid. This study has delineated the different stomach miRNAs expression patterns of Tibetan and Yorkshire pigs, which would help explain the regulatory mechanisms of miRNAs in digestion of Tibetan pigs, and contribute to utilize a the unique digestion merits of Tibetan pig in future porcine hybridization breeding.

Rheinheimera aquatica sp. nov., Antimicrobial Activity-Producing Bacterium Isolated from Freshwater Culture Pond

( Wen Ming Chen ),( Yi Lin Chang ),( Chung Young Chiu ),( Yi Sheu Shih ) 한국미생물 · 생명공학회 2010 Journal of microbiology and biotechnology Vol.20 No.10

A bacterial strain designated GR5T, previously isolated from a freshwater culture pond in Taiwan while screening for bacteria for antimicrobial compounds, was characterized using a polyphasic taxonomic approach. Strain GR5T was found to be Gram-negative, aerobic, greenish-yellow colored, rod-shaped, and motile by means of a single polar flagellum. Growth occurred at 10-40oC (optimum, 35oC), pH 7.0-8.0 (optimum pH 8.0), and with 0-2.0% NaCl (optimum, 0.5-1.0%). The major fatty acids were C16:1ω7c (36.3%), C16:0 (16.6%), C12:0 3-OH (12.5%), and C18:1ω7c (9.1%). The major respiratory quinone was Q-8, and the DNA G+C content of the genomic DNA was 51.9 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain GR5T belongs to the genus Rheinheimera, where its most closely related neighbors are Rheinheimera texasensis A62-14BT and Rheinheimera tangshanensis JA3-B52T with sequence similarities of 98.1% and 97.5%, respectively, and the sequence similarities to any other recognized species within Gammaproteobacteria are less than 96.5%. The mean level of DNA-DNA relatedness between strain GR5 T and R. texasensis A62-14BT, the strain most closely related to the isolate, was 26.5±7.6%. Therefore, based on the phylogenetic and phenotypic data, strain GR5T should be classified as a novel species, for which the name Rheinheimera aquatica sp. nov. is proposed. The type strain is GR5T (=BCRC 80081T= LMG 25379T).

Anti-colonic Inflammation by Black Raspberries through Regulating Toll-like Receptor-4 Signaling in Interlukin-10 Knockout Mice

Yi-Wen Huang,Carla Elena Echeveste,Kiyoko Oshima,Jianying Zhang,Martha Yearsley,Jianhua Yu,Li-Shu Wang 대한암예방학회 2020 Journal of cancer prevention Vol.25 No.2

Ulcerative colitis (UC) is a chronic inflammatory disease of the colon, with a steadily rising prevalence in Western and newly industrialized countries. UC patients have a cancer incidence as high as 10% after 20 years of the disease. Although the importance of fruits and vegetables in defense against UC is beginning to be appreciated, the mechanisms remain largely unclear. In the current study, we reported that dietary black raspberries (BRBs) decreased colonic inflammation in the mucosa and submucosa of interleukin (IL)-10 knockout (KO) mice. We then used colon, spleen, and plasma from those mice to investigate whether BRBs exert their anti-inflammatory effects by correcting dysregulated toll-like receptor (TLR)-4 signaling to downregulate prostaglandin E2 (PGE2). Other studies reported that spleen is the reservoir of macrophages and depletion of macrophages in IL-10 KO mice prevents the development of colitis. Our results showed that BRBs decreased the percentages of macrophages in spleens of IL-10 KO mice. Moreover, mechanistically, the BRB diet corrected dysregulated TLR-4 signaling in cells from the colon and spleen, decreased PGE2 and prostaglandin I2, and increased 15-lipoxygenase and its product, 13-S-hydroxyoctadecadienoic acid, in plasma of IL- 10 KO mice. Therefore, we have elucidated one of the anti-inflammatory mechanisms of BRBs, and have identified biomarkers that could be indicators of response in UC patients treated with them. Our findings with BRBs could well apply to many other commonly consumed fruits and vegetables.

PF2405, standardized fraction of Scutellaria baicalensis, ameliorates colitis in vitro and in vivo

Wen-Yi Jiang,서검석,김윤철,손동환,이성희 대한약학회 2015 Archives of Pharmacal Research Vol.38 No.6

Standardized extraction procedures for herb areas important as their authentication to maintain theirquality and ensure their safe use. We had prepared astandardized and purified Scutellaria baicalensis Georgiextract, PF2405, which was enriched with three majorcomponents, baicalein, oroxylin A and wogonin. In thepresent study, we investigated the potential anti-inflammatoryeffects of PF2405 in vitro and in two differentexperimental animal models of inflammatory bowel disease. Effect of PF2405 studied in tumor necrosis factor(TNF)-a-induced HT-29 cells in vitro. In vivo experimentalcolitis models were induced by administration oftrinitrobenzene sulfonic acid (TNBS) or dextran sulfatesodium (DSS). PF2405 (50 lg/ml) decreased TNF-ainducedcyclooxygenase (COX)-2 expressions throughinhibition of phosphorylation of c-Jun N-terminal kinasesand p38 mitogen-activated protein kinase in HT-29 cells. Combination of baicalein (20 lg/ml), oroxylin A (8 lg/ml),and wogonin (2 lg/ml) markedly inhibits TNF-a-inducedCOX-2 expression when compared with individual components. PF2405 (25 mg/kg b.w.) treatment significantlyreduced histopathological severity; suppressed expression ofCOX-2, TNF-a, and interleukin-1b in TNBS-induced mice. Moreover, PF2405 (25 mg/kg b.w.) has both potent preventiveand therapeutic activities in DSS-induced colitis. Collectively, PF2405 shows prominent anti-inflammatoryeffect that can be used as a new therapeutic approach forintestinal inflammatory disorders.

Wave propagation and vibration of FG pipes conveying hot fluid

Yi-Wen Zhang,Gui-Lin She 국제구조공학회 2022 Steel and Composite Structures, An International J Vol.42 No.3

The existing researches on the dynamics of the fluid-conveying pipes only focus on stability and vibration problems, and there is no literature report on the wave propagation of the fluid-conveying pipes. Therefore, the purpose of this paper is to explore the propagation characteristics of longitudinal and flexural waves in the fluid-conveying pipes. First, it is assumed that the material properties of the fluid-conveying pipes vary based on a power function of the thickness. In addition, it is assumed that the material properties of both the fluid and the pipes are closely depended on temperature. Using the Euler-Bernoulli beam equation and based on the linear theory, the motion equations considering the thermal-mechanical-fluid coupling is derived. Then, the exact expressions of phase velocity and group velocity of longitudinal waves and bending waves in the fluidconveying pipes are obtained by using the eigenvalue method. In addition, we also studied the free vibration frequency characteristics of the fluid-conveying pipes. In the numerical analysis, we successively studied the influence of temperature, functional gradient index and liquid velocity on the wave propagation and vibration problems. It is found that the temperature and functional gradient exponent decrease the phase and group velocities, on the contrary, the liquid flow velocity increases the phase and group velocities. However, for vibration problems, temperature, functional gradient exponent parameter, and fluid velocity all reduce the natural frequency.

Antioxidant Activity of Cleomiscosins A and C Isolated from Acer okamotoanum

Wen Yi Jin,Phuong Thien Thuong,Nguyen Duy Su,민병선,손건호,장현욱,김현표,강삼식,석대은,배기환 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.3

Phytochemical investigation of Acer okamotoanum leaf and twig led to the isolation of two coumarinolignans, cleomiscosin A (1) and cleomiscosin C (2). Here, we found that 2 dose-dependently inhibits LDL oxidation mediated by either catalytic copper ions (Cu2+) or free radicals generated with the azo compound 2,2'-azobis-(2-amidinopropane)dihydro-chloride (AAPH) with IC50s of 29.5 and 11.9 µM, respectively. By electrophoretic analysis, we also observed that 2 protects apolipoprotein B-100 (apoB-100) against Cu2+-induced fragmentation (65.3% inhibition at 5 µM). Furthermore, fluorescence analyses clearly indicated that both 1 and 2 protect against the oxidative modification of apoB-100 induced by either Cu2+ or HOCl (1, IC50s of 13.4 and 8.1 µM, respectively; 2, IC50s of 23.6 and 3.9 µM, respectively). These findings suggest that 1 and 2 could be beneficial in preventing LDL oxidation in atherosclerotic lesions.

Effects of Modafinil on Behavioral Learning and Hippocampal Synaptic Transmission in Rats

Wen-Wen Yan,Li-Hua Yao,Chong Chen,Hai-Xia Wang,Chu-Hua Li,Jun-Ni Huang,Peng Xiao,Cheng-Yi Liu 대한배뇨장애요실금학회 2015 International Neurourology Journal Vol.19 No.4

Purpose: Modafinil is a wake-promoting agent that has been proposed to improve cognitive performance at the preclinical and clinical levels. Since there is insufficient evidence for modafinil to be regarded as a cognitive enhancer, the aim of this study was to investigate the effects of chronic modafinil administration on behavioral learning in healthy adult rats. Methods: Y-maze training was used to assess learning performance, and the whole-cell patch clamp technique was used to assess synaptic transmission in pyramidal neurons of the hippocampal CA1 region of rats. Results: Intraperitoneal administration of modafinil at 200 mg/kg or 300 mg/kg significantly improved learning performance. Furthermore, perfusion with 1mM modafinil enhanced the frequency and amplitude of spontaneous postsynaptic currents and spontaneous excitatory postsynaptic currents in CA1 pyramidal neurons in hippocampal slices. However, the frequency and amplitude of spontaneous inhibitory postsynaptic currents in CA1 pyramidal neurons were inhibited by treatment with 1mM modafinil. Conclusions: These results indicate that modafinil improves learning and memory in rats possibly by enhancing glutamatergic excitatory synaptic transmission and inhibiting GABAergic (gamma-aminobutyric acid-ergic) inhibitory synaptic transmission.

Surveillance of the progression and assessment of treatment endpoints for nonalcoholic steatohepatitis

Yi-Wen Shi,Jian-Gao Fan 대한간학회 2023 Clinical and Molecular Hepatology(대한간학회지) Vol.29 No.-

Nonalcoholic steatohepatitis (NASH) is an aggressive form of nonalcoholic fatty liver disease (NAFLD) characterized by steatosis-associated inflammation and liver injury. Without effective treatment or management, NASH can have life-threatening outcomes. Evaluation and identification of NASH patients at risk for adverse outcomes are therefore important. Key issues in screening NASH patients are the assessment of advanced fibrosis, differentiation of NASH from simple steatosis, and monitoring of dynamic changes during follow-up and treatment. Currently, NASH staging and evaluation of the effectiveness for drugs still rely on pathological diagnosis, despite sample error issues and the subjectivity associated with liver biopsy. Optimizing the pathological assessment of liver biopsy samples and developing noninvasive surrogate methods for accessible, accurate, and safe evaluation are therefore critical. Although noninvasive methods including elastography, serum soluble biomarkers, and combined models have been implemented in the last decade, noninvasive diagnostic measurements are not widely applied in clinical practice. More work remains to be done in establishing cost-effective strategies both for screening for at-risk NASH patients and identifying changes in disease severity. In this review, we summarize the current state of noninvasive methods for detecting steatosis, steatohepatitis, and fibrosis in patients with NASH, and discuss noninvasive assessments for screening at-risk patients with a focus on the characteristics that should be monitored at follow-up.

Dysregulated Free Fatty Acid Receptor 2 Exacerbates Colonic Adenoma Formation in ApcMin/+ Mice: Relation to Metabolism and Gut Microbiota Composition

Yi-Wen Huang,Chien-Wei Lin,Pan Pan,Carla Elena Echeveste,Athena Dong,Kiyoko Oshima,Martha Yearsley,Jianhua Yu,Li-Shu Wang 대한암예방학회 2021 Journal of cancer prevention Vol.26 No.1

Free fatty acid receptor 2 (FFAR2) has been reported as a tumor suppressor in colon cancer development. The current study investigated the effects of FFAR2 signaling on energy metabolism and gut microbiota profiling in a colorectal cancer mouse model (ApcMin/+). Ffar2 deficiency promoted colonic polyp development and enhanced fatty acid oxidation and bile acid metabolism. Gut microbiome sequencing analysis showed distinct clustering among wild-type, ApcMin/+, and ApcMin/+-Ffar2-/- mice. The relative abundance of Flavobacteriaceae and Verrucomicrobiaceae was significantly increased in the ApcMin/+-Ffar2-/- mice compared to the ApcMin/+ mice. In addition, knocking-down FFAR2 in the human colon cancer cell lines (SW480 and HT29) resulted in increased expression of several key enzymes in fatty acid oxidation, such as carnitine palmitoyltransferase 2, acyl-CoA dehydrogenase, longchain acyl-CoA dehydrogenase, C-2 to C-3 short chain, and hydroxyacyl-CoA dehydrogenase/3-ketoacyl-CoA thiolase/enoyl-CoA hydratase, alpha subunit. Collectively, these results demonstrated that Ffar2 deficiency significantly altered profiles of fatty acid metabolites and gut microbiome, which might promote colorectal cancer development.