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.

Combined Assessment of Serum Alpha-Synuclein and Rab35 is a Better Biomarker for Parkinson’s Disease

Hung-Li Wang,Chin-Song Lu,Tu-Hsueh Yeh,Yu-Ming Shen,Yi-Hsin Weng,Ying-Zu Huang,Rou-Shayn Chen,Yu-Chuan Liu,Yi-Chuan Cheng,Hsiu-Chen Chang,Ying-Ling Chen,Yu-Jie Chen,Yan-Wei Lin,Chia Chen Hsu,Huang-Li 대한신경과학회 2019 Journal of Clinical Neurology Vol.15 No.4

Background and Purpose It is essential to develop a reliable predictive serum biomarker for Parkinson’s disease (PD). Te accumulation of alpha-synuclein (αSyn) and up-regulated expression of Rab35 participate in the etiology of PD. Te purpose of this investigation was to determine whether the combined assessment of serum αSyn and Rab35 is a useful predictive biomarker for PD. Methods Serum levels of αSyn or Rab35 were determined in serum samples from 59 sporadic PD patients, 19 progressive supranuclear palsy (PSP) patients, 20 multiple system atrophy (MSA) patients, and 60 normal controls (NC). Receiver operating characteristics (ROC) curves were calculated to determine the diagnostic accuracy of αSyn or/and Rab35 in discriminating PD patients from NC or atypical parkinsonian patients. Results The levels of αSyn and Rab35 were increased in PD patients. The serum level of Rab35 was positively correlated with that of αSyn in PD patients. Compared to analyzing αSyn or Rab35 alone, the combined analysis of αSyn and Rab35 produced a larger area under the ROC curve and performed better in discriminating PD patients from NC, MSA patients, or PSP patients. When age was dichotomized at 55, 60, 65, or 70 years, the combined assessment of αSyn and Rab35 for classifying PD was better in the group below the cutof age than in the group above the cutof age. Conclusions Combined assessment of serum αSyn and Rab35 is a better biomarker for discriminating PD patients from NC or atypical parkinsonian patients, and is a useful predictive biomarker for younger sporadic PD patients.

Risk Factors for Anxiety in Major Depressive Disorder Patients

Li-Min Xin,Lin Chen,Zhen-Peng Ji,Suo-Yuan Zhang,Jun Wang,Yan-Hong Liu,Da-Fang Chen,Fu-De Yang,Gang Wang,Yi-Ru Fang,Zheng Lu,Hai-Chen Yang,Jian Hu,Zhi-Yu Chen,Yi Huang,Jing Sun,Xiao-Ping Wang,Hui-Chun 대한정신약물학회 2015 CLINICAL PSYCHOPHARMACOLOGY AND NEUROSCIENCE Vol.13 No.3

Objective: To analyze the sociodemographic and clinical factors related to anxiety in patients with major depressive disorder (MDD). Methods: This study involved a secondary analysis of data obtained from the Diagnostic Assessment Service for People with Bipolar Disorders in China (DASP), which was initiated by the Chinese Society of Psychiatry (CSP) and conducted from September 1, 2010 to February 28, 2011. Based on the presence or absence of anxiety-related characteristics, 1,178 MDD patients were classified as suffering from anxious depression (n=915) or non-anxious depression (n=263), respectively. Results: Compared with the non-anxious group, the anxious-depression group had an older age at onset (t=−4.39, p<0.001), were older (t=−4.69, p<0.001), reported more lifetime depressive episodes (z=−3.24, p=0.001), were more likely to experience seasonal depressive episodes (χ2=6.896, p=0.009) and depressive episodes following stressful life events (χ2=59.350, p <0.001), and were more likely to have a family history of psychiatric disorders (χ2=6.091, p=0.014). Their positive and total scores on the Mood Disorder Questionnaire (MDQ) and the 32-item Hypomania Checklist (HCL-32) (p<0.05) were also lower. The logistic regression analysis indicated that age (odds ratio [OR]=1.03, p<0.001), a lower total MDQ score (OR=0.94, p=0.011), depressive episodes following stressful life events (OR=3.04, p<0.001), and seasonal depressive episodes (OR=1.75, p=0.039) were significantly associated with anxious depression. Conclusion: These findings indicate that older age, fewer subclinical bipolar features, an increased number of depressive episodes following stressful life events, and seasonal depressive episodes may be risk factors for anxiety-related characteristics in patients with MDD.

Prognostic Significance of α5β1-integrin Expression in Cervical Cancer

Wang, Hua-Yi,Chen, Zhe,Wang, Zhu-Hui,Wang, Hong,Huang, Li-Ming Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.6

The purpose of this study was to evaluate the association of expression of ${\alpha}5{\beta}1$-integrin with clinicopathologic features and prognosis in cervical cancer. Levels of ${\alpha}5{\beta}1$-integrin in normal cervical mucosa and cervical cancer tissue were detected with immunohistochemistry. Survival analysis by the Kaplan-Meier method was performed to assess prognostic significance. ${\alpha}5{\beta}1$-integrin expression was detected in 84.6% (143/169) cervical cancer samples, significantly different from that in normal cervical mucosa (P < 0.05). Positive expression rates of ${\alpha}5{\beta}1$-integrin in patients with poor histologic differentiation, lymph node metastasis, and recurrence were elevated. Using Kaplan-Meier analysis, a comparison of survival curves of low versus high expression of ${\alpha}5{\beta}1$-integrin revealed a highly significant difference in human cervical cancer cases (P < 0.05), suggesting that overexpression of ${\alpha}5{\beta}1$-integrin is associated with a worse prognosis.The ${\alpha}5{\beta}1$-integrin promotes angiogenesis and associates with lymph node metastasis, vascular invasion and poor prognosis of cervical cancer. The current study indicated that ${\alpha}5{\beta}1$-integrin may be an independent prognostic factor for cervical cancer patients.

Temperature dependent development of black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae) from Yangtze River Delta region of China

Wang Yinghui,Zhang Yanan,Wang Jiangfeng,Kang Chengtao,Hu Gengwang,Guo Yi,Chen Jie,Yang Lijun,Wang Yu 한국응용곤충학회 2023 Journal of Asia-Pacific Entomology Vol.26 No.4

The black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae), isomnivorous andwidely distributed in the world. Black soldier flies usually colonize corpse at a relatively late stage and have a longer developmental time compared to other flies. It can serve as a good supplementary indicator for estimating a longer minimum postmortem interval (PMI min ) in forensic investigations. In this paper, the development of H. illucens was investigated at six constant temperatures between 19 and 34 ◦ C. Under temperatures of 19, 22, 25, 28, 31, and 34 ◦ C, the mean developmental time from eggs to adults was 90.96, 58.29, 54.61, 42.33, 32.33 and 34.52 days, respectively. Developmental time and accumulated degree days results were used to construct thermal sum mation model and isomorphen diagram. The calculated values of lower developmental thresholds and thermal summation constant estimated by a linear model were 11.88 ◦ C and 650.89 degree days, respectively. Through a nonlinear model, the upper lethal developmental threshold temperature, intrinsic optimum temperature and lower developmental thresholds were estimated to be 35.18, 20.50 and 13.17 ◦ C, respectively. A growth curve, an equation and isomegalen diagram based on them show the relationship between developmental time and larval body length. The larval body length increased with increased developmental time and eventually pla teaued, revealing an S-shape growth curve. These results provide important basic developmental data for H. illucens, which can be used to estimate the PMI min .

Freeze-Casting Produces a Graphene Oxide Aerogel with a Radial and Centrosymmetric Structure

Wang, Chunhui,Chen, Xiong,Wang, Bin,Huang, Ming,Wang, Bo,Jiang, Yi,Ruoff, Rodney S. American Chemical Society 2018 ACS NANO Vol.12 No.6

<P>We report the assembly of graphene oxide (G-O) building blocks into a vertical and radially aligned structure by a bidirectional freeze-casting approach. The crystallization of water to ice assembles the G-O sheets into a structure, a G-O aerogel whose local structure mimics turbine blades. The centimeter-scale radiating structure in this aerogel has many channels whose width increases with distance from the center. This was achieved by controlling the formation of the ice crystals in the aqueous G-O dispersion that grew radially in the shape of lamellae during freezing. Because the shape and size of ice crystals is influenced by the G-O sheets, different additives (ethanol, cellulose nanofibers, and chitosan) that can form hydrogen bonds with H<SUB>2</SUB>O were tested and found to affect the interaction between the G-O and formation of ice crystals, producing ice crystals with different shapes. A G-O/chitosan aerogel with a spiral pattern was also obtained. After chemical reduction of G-O, our aerogel exhibited elasticity and absorption capacity superior to that of graphene aerogels with “traditional” pore structures made by conventional freeze-casting. This methodology can be expanded to many other configurations and should widen the use of G-O (and reduced G-O and “graphenic”) aerogels.</P> [FIG OMISSION]</BR>

The GntR-Type Regulators GtrA and GtrB Affect Cell Growth and Nodulation of Sinorhizobium meliloti

Yi Wang,Ai-Min Chen,Ai-Yuan Yu,Li Luo,Guan-Qiao Yu,Jia-Bi Zhu,Yan-Zhang Wang 한국미생물학회 2008 The journal of microbiology Vol.46 No.2

GntR-type transcriptional regulators are involved in the regulation of various biological processes in bacteria, but little is known about their functions in Sinorhizobium meliloti. Here, we identified two GntR-type transcriptional regulator genes, gtrA and gtrB, from S. meliloti strain 1021. Both the gtrA1 mutant and the gtrB1 mutant had lower growth rates and maximal cell yields on rich and minimal media, as well as lower cell motility on swimming plates, than did the wild-type strain. Both mutants were also symbiotically deficient. Alfalfa plants inoculated with wild-type strain 1021 formed pink elongated nodules on primary roots. In contrast, the plants inoculated with the gtrA1 and gtrB1 mutants formed relatively smaller, round, light pink nodules mainly on lateral roots. During the first 3~4 weeks post-inoculation, the plants inoculated with the gtrA1 and gtrB1 mutants were apparently stunted, with lower levels of nitrogenase activity, but there was a remarkable increase in the number of nodules compared to those inoculated with the wild-type strain. Moreover, the gtrA1 and gtrB1 mutants not only showed delayed nodulation, but also showed markedly reduced nodulation competition. These results demonstrated that both GtrA and GtrB affect cell growth and effective symbiosis of S. meliloti. Our work provides new insight into the functions of GntR-like transcriptional regulators.

Characterization and Functional Analysis of Bacillus aryabhattai CY for Acrylic Acid Biodegradation: Immobilization and Metabolic Pathway

Yi Chen,Min Zhao,Liyong Hu,Zeyu Wang,Dzmitry Hrynsphan,Jun Chen 한국생물공학회 2021 Biotechnology and Bioprocess Engineering Vol.26 No.6

Acrylic acid has been widely used in various industrial applications but is harmful to human health and the environment. A novel and efficient degrading acrylic acid bacterium was isolated and identified as Bacillus aryabhattai CY. In this study, batch experiments were conducted to evaluate the biodegradation of acrylic acid by B. aryabhattai CY, which were immobilized in calciumalginate beads under different conditions. The components of the alginate beads were optimized by the response surface method, and the degradation performance of the immobilized cells was determined. Relative to the free cells, experiment results showed that the immobilized cells can achieve complete degradation of 100 mg/L acrylic acid in 24 h under the optimal conditions of SA 6% (w/v), CaCl2 1% (w/v), and immobilization time of 6 h. According to Haldane's model, the maximum specific growth rate (μmax) of the free cells and immobilized cells were 0.165/h and 0.210/h, respectively. Experiment data revealed that acrylic acid showed an inhibitory effect on biodegradation by B. aryabhattai CY, especially at concentration higher than 100 mg/L. Furthermore, the reusability of the immobilized cells revealed that the acrylic acid removal rate was above 93.70% within the eight cycles. The immobilized cells also showed higher removal efficiencies in wider ranges of temperature (20°C-60°C) and pH (5.0-10.0) than the free cells. Moreover, the possible degradation intermediates were proposed during the biodegradation of acrylic acid by GC-MS analysis. Results indicated that immobilized beads might have a potential environmental implication in the purification of practical acrylic acid wastewater.

Notch1 promotes the pericytemyofibroblast transition in idiopathic pulmonary fibrosis through the PDGFR/ ROCK1 signal pathway

Yi-Chun Wang,Qiong Chen,Jun-Ming Luo,Jing Nie,Qing-He Meng,Wei Shuai,Han Xie,Jia-Mei Xia,Hui Wang 생화학분자생물학회 2019 Experimental and molecular medicine Vol.51 No.-

The goals of this study were to investigate the role of the Notch1/PDGFRβ/ROCK1 signaling pathway in the pathogenesis of pulmonary fibrosis and to explore the possibility of treating fibrosis by targeting Notch1. Lung tissues from patients with pulmonary fibrosis were examined for the expression of Notch1/PDGFRβ/ROCK1 using RT-qPCR, western blotting, and immunostaining. Cultured mouse lung pericytes were transfected with Notch1-overexpressed vectors or shRNA targeting PDGFRβ/ROCK1 to examine cell behaviors, including proliferation, cell cycle arrest, and differentiation toward myofibroblasts. Finally, a mouse pulmonary fibrosis model was prepared, and a Notch1 inhibitor was administered to observe tissue morphology and pericyte cell behaviors. Human pulmonary fibrotic tissues presented with overexpression of Notch1, PDGFRβ, and ROCK1, in addition to a prominent transition of pericytes into myofibroblasts. In cultured mouse lung pericytes, overexpression of Notch1 led to the accelerated proliferation and differentiation of cells, and it also increased the expression of the PDGFRβ and ROCK1 proteins. The knockdown of PDGFRβ/ROCK1 in pericytes remarkably suppressed pericyte proliferation and differentiation. As further substantiation, the administration of a Notch1 inhibitor in a mouse model of lung fibrosis inhibited the PDGFRβ/ROCK1 pathway, suppressed pericyte proliferation and differentiation, and alleviated the severity of fibrosis. Our results showed that the Notch1 signaling pathway was aberrantly activated in pulmonary fibrosis, and this pathway may facilitate disease progression via mediating pericyte proliferation and differentiation. The inhibition of the Notch1 pathway may provide one promising treatment strategy for pulmonary fibrosis.

Influence of Ga(Al)As substrates on surface morphology and critical thickness of InGaAs quantum dots

Yi Wang,Xiang Guo,Jiemin Wei,Chen Yang,Zijiang Luo,Jihong Wang,Zhao Ding 한국물리학회 2019 Current Applied Physics Vol.19 No.4

Influence of Ga(Al)As substrates on surface morphology of InGaAs quantum dots and critical thickness of In0.5Ga0.5As film grown by molecular beam epitaxy is investigated. The In0.5Ga0.5As quantum dots are grown on (001) surfaces of GaAs and Al0.25Ga0.75 A at 450 °C, scanning tunneling microscope images show that the size of quantum dots varied slightly for 10 ML of In0.5Ga0.5As grown on GaAs and Al0.25Ga0.75As surfaces. Reflection high energy electron diffraction (RHEED) is used to monitor the growth of 4 monolayers (ML) In0.5Ga0.5As on Al0.25Ga0.75As and GaAs surfaces during deposition. The critical thickness is theoretically calculated by adding energy caused by surface roughness and heat from substrate. The calculations show that the critical thickness of In0.5Ga0.5As grown on GaAs and Al0.25Ga0.75As are 3.2 ML and 3.8 ML, respectively. The theoretical calculation agrees with the experimental results.