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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.
Lytic Bacteriophage as a Biomaterial to Prevent Biofilm Formation and Promote Neural Growth
Liu Zi-Hao,Chiang Ming-Tse,Lin Hsin-Yi 한국조직공학과 재생의학회 2022 조직공학과 재생의학 Vol.19 No.5
BACKGROUND Although non-lytic filamentous bacteriophages have been made into biomaterial to guide tissue growth, they had limited ability to prevent bacterial infection. In this work a lytic bacteriophage was used to make an antibacterial biomaterial for neural tissue repair. METHODS Lytic phages were chemically bound to the surface of a chitosan film through glutaraldehyde crosslinking. After the chemical reaction, the contact angle of the sample surface and the remaining lytic potential of the phages were measured. The numbers of bacteria on the samples were measured and examined under scanning electron microscopy. Transmission electron microscopy (TEM) was used to observe the phages and phage-infected bacteria. A neuroblast cell line was cultured on the samples to evaluate the sample’s biocompatibility. RESULTS The phages conjugated to the chitosan film preserved their lytic potential and reduced 68% of bacterial growth on the sample surface at 120 min (p\0.001). The phage-linked surface had a significantly higher contact angle than that of the control chitosan (p\0.05). After 120 min a bacterial biofilm appeared on the control chitosan, while the phagelinked sample effectively prevented biofilm formation. The TEM images demonstrated that the phage attached and lysed the bacteria on the phage-linked sample at 120 min. The phage-linked sample significantly promoted the neuroblast cell attachment (p\0.05) and proliferation (p\0.01). The neuroblast on the phage-linked sample demonstrated more cell extensions after day 1. CONCLUSION The purified lytic phages were proven to be a highly bioactive nanomaterial. The phage-chitosan composite material not only promoted neural cell proliferation but also effectively prevent bacterial growth, a major cause of implant failure and removal.
Yi-Chih Chang,Hao-Ping Liu,Hsiao-Li Chuang,Jiunn-Wang Liao,Pei-Ling Kao,Hsun-Lung Chan,Ter-Hsin Chen,Yu-Chih Wang 한국실험동물학회 2023 Laboratory Animal Research Vol.39 No.4
Background: Feline mammary carcinoma (FMC) is one of the most prevalent malignancies of female cats. FMC is highly metastatic and thus leads to poor disease outcomes. Among all metastases, liver metastasis occurs in about 25% of FMC patients. However, the mechanism underlying hepatic metastasis of FMC remains largely uncharacterized. Results: Herein, we demonstrate that FMC-derived extracellular vesicles (FMC-EVs) promotes the liver metastasis of FMC by activating hepatic stellate cells (HSCs) to prime a hepatic premetastatic niche (PMN). Moreover, we provide evidence that sphingosine kinase 1 (SK1) delivered by FMC-EV was pivotal for the activation of HSC and the formation of hepatic PMN. Depletion of SK1 impaired cargo sorting in FMC-EV and the EV-potentiated HSC activation, and abolished hepatic colonization of FMC cells. Conclusions: Taken together, our findings uncover a previously uncharacterized mechanism underlying liver-metastasis of FMC and provide new insights into prognosis and treatment of this feline malignancy.
Piperacillin-Tazobactam-Induced Linear IgA Bullous Dermatosis Supported by a T-Cell Activation Assay
( Yi-hsin Ho ),( Yun-wen Chiu ),( Han-nan Liu ) 대한피부과학회 2018 Annals of Dermatology Vol.30 No.5
Linear immunoglobulin (Ig) A bullous dermatosis (LABD) is a rare subepidermal autoimmune blistering disease characterized by linear IgA deposits at the basement membrane zone visualized with direct immunofluorescence (DIF). Most cases of LABD are idiopathic, but some are drug-induced with vancomycin being the most common causative agent. We herein report a patient presenting with blisters and erosive lesions, primarily in the intertriginous and flexor areas, consistent with a diagnosis of piperacillin-tazobactam-induced LABD based on the patient’s clinical course and histopathology, DIF, and in vitro T-cell activation assay (TAA) findings. Only one case of piperacillin-tazobactam-induced LABD has been previously reported. In addition to its rarity, our case was also unique in that the skin lesions occurred in the intertriginous and flexor areas, uncommon locations for typical adult patients with LABD, and TAA strongly suggested an association with the causative drug. (Ann Dermatol 30(5) 588∼591, 2018)
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%.
Wan-Hsin Chang,Pei-Yi Liu,Dai-En Lin,Yuan-Ting Jiang,Chien-Ju Lu,Yuan-Hao Howard Hsu 한국고분자학회 2022 Macromolecular Research Vol.30 No.1
Adsorption of secreted protein on contact lenses is a dynamic and complex behavior. To understand this behavior, we used three-dimensional (3D) printing technology to create an eye model that simulated the anterior segment of the actual human eyeball. In this model, the fluid inlet was connected to a syringe pump to mimic the rate of human tear secretion and the outlet was connected to an ultraviolet- visible (UV-Vis) spectrophotometer. The experimental results revealed that the symmetrical eye model with a 180° inlet-outlet angle was suitable for dynamic analysis of protein adsorption. In this model, protein adsorption was slow and desorption was rapid. The contact lens was soaked in poly(2-methacryloyloxyethyl phosphorylcholine- co-butyl methacrylate) (PMB) to confirm the anti-protein adsorption property of this polymer through dynamic adsorption and desorption eye model analysis.
( Wen Fu Wu ),( Yi Ching Liu ),( Hsin Yun Chang ),( Chen Wei Huang ),( Ying Chen Lu ) 한국농업기계학회 2018 한국농업기계학회 학술발표논문집 Vol.23 No.1
Pineapples are tropical fruit that are rich in vitamins, enzymes and fibers with many health benefits not only in pulps but also peels. However, the peels of pineapples are normally removed and dropped. The pulps and peels of four pineapple cultivars, Milk, Honey Fragrance, Smooth Cayenne and Golden Diamond pineapples, were juiced, filtered and analyzed, and the Golden Diamond pineapple was found to have highest antioxidant capacity. The pulp and peel juices of Golden Diamond pineapple were inoculated with Enterococcus faecium LYC91 and various concentration calcium carbonate to ferment at 37°C for 24 hours follow by bacterial and antioxidant capacity analysis. The results indicated that the fermented pineapple peel juice with calcium carbonate had highest bacterial number, total polyphenol content, DPPH radical scavenging ability, Fe2 + chelating ability and reducing power activity than pulp juice and unfermented pineapple juice. The viable bacterial number of fermented pineapple pulp and peel juices with calcium carbonate remain 8.0 log CFU/mL after stored at 4°C for 1 month. The freeze-dried powder of the fermented pineapple pulp and peel juices with calcium carbonate retained the viable cell number of 9.0 log CFU/g after stored at 4°C for three months. These results showed that the fermented pineapple pulp and peel juices with calcium carbonate were novel pineapple lactic acid fermented drinks with growth potential.
Sheng-Fuu Lin,Jau-Chi Ke,Hsin-Yi Liu,Jiao-Rou Liao 제어로봇시스템학회 2014 제어로봇시스템학회 국제학술대회 논문집 Vol.2014 No.10
In this paper, a new method for three-dimensional measurement with two horizontally-aligned omni-directional cameras is proposed. Some omni-camera stereo vision systems calculate three-dimensional information which always considers either azimuth angles or elevation angles causing inaccuracy easily. The proposed method takes into account both azimuth angles and elevation angles in order to reduce errors. Besides, the range of interest (ROI) is 3 to 8 meters around cameras in the system. The proposed method has better performance than the vertically- or horizontally-aligned in the 3D measurement.