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Polymerase engineering

Jeong Eun Hyeon, Da Woon Jeong, Young Jin Ko, Seung Wook Kim, Chulhwan Park, Sung Ok Han
Regardless of the malaria disease risk, the malaria parasite Plasmodium falciparum has an interesting mechanism. During its growth within the red blood cell, toxic free heme is converted into an insoluble crystalline form called the malaria pigment, or hemozoin. In particular, natural hemozoin nanocrystals can provide multiple applications in biosensing fields for health care and diagnosis as similar to artificial metal nanoparticles. In this study, the heme was biologically synthesized and polymerized by Corynebacterium glutamicum and final polymer was applied as a biomimetic conductive biopolymer...
May 10, 2018: Biosensors & Bioelectronics
Bing-Bing Li, Yuan-Yuan Cheng, Yang-Yang Fan, Dong-Feng Liu, Cai-Yun Fang, Chao Wu, Wen-Wei Li, Zong-Chuang Yang, Han-Qing Yu
Shewanella species have a diverse respiratory ability and wide distribution in environments and play an important role in bioremediation and the biogeochemical cycles of elements. Primers with more accuracy and broader coverage are required with consideration of the increasing number of Shewanella species and evaluation of their roles in various environments. In this work, a new primer set of 640F/815R was developed to quantify the abundance of Shewanella species in natural and engineered environments. In silico tools for primer evaluation, quantitative polymerase chain reaction (qPCR) and clone library results showed that 640F/815R had a higher specificity and coverage than the previous primers in quantitative analysis of Shewanella...
May 12, 2018: Science of the Total Environment
Federico Mussano, Tullio Genova, Sara Petrillo, Ilaria Roato, Riccardo Ferracini, Luca Munaron
Great efforts have been made to improve bone regeneration techniques owing to a growing variety of sources of stem cells suitable for autologous transplants. Specifically, adipose-derived stem cells (ASCs) and stems cells from human exfoliated deciduous teeth (SHED) hold great potential for bone tissue engineering and cell therapy. After a preliminary characterization of the main biomolecules ASCs and SHED released in their conditioned media, cells were kept both in normal and osteo-inducing conditions. Conventional assays were performed to prove their osteogenic potential such as quantitative real-time polymerase chain reaction (qRT-PCR) (for RUNX-2, collagen type I, osteopontin and osteonectin), alkaline phosphatase activity, osteocalcin production, and von Kossa staining...
May 14, 2018: International Journal of Molecular Sciences
Zhen Kang, Wenwen Ding, Peng Jin, Guocheng Du, Jian Chen
In past decades, DNA engineering protocols have led to the rapid development of synthetic biology. To engineer the natural proteins, many directed evolution methods based on molecular biology have been presented for generating genetic diversity or obtaining specific properties. Here, we provide a simple (PCR operation), efficient (larger amount of products), and powerful (multiple point mutations, deletions, insertions, and combinatorial multipoint mutagenesis) RECODE method, which is capable of reediting the target DNA flexibly to restructure regulatory regions and remodel enzymes by using the combined function of the thermostable DNA polymerase and DNA ligase in one pot...
2018: Methods in Molecular Biology
Xun Sun, Yun Zhu, He-Yong Yin, Zhi-Yuan Guo, Feng Xu, Bo Xiao, Wen-Li Jiang, Wei-Min Guo, Hao-Ye Meng, Shi-Bi Lu, Yu Wang, Jiang Peng
BACKGROUND: Peripheral nerve injury (PNI) is a worldwide issue associated with severe social and economic burden. Autologous nerve grafting, the gold standard treatment for peripheral nerve defects, still has a number of technical limitations. Tissue engineering technology is a novel therapeutic strategy, and mesenchymal stromal cells (MSCs) are promising seed cells for nerve tissue engineering. However, the efficiency of traditional methods for inducing the differentiation of MSCs to Schwann cell-like cells (SCLCs) remains unsatisfactory...
May 11, 2018: Stem Cell Research & Therapy
Se-Joon Oh, Hee-Young Park, Kyung-Un Choi, Sung-Won Choi, Sung-Dong Kim, Soo-Keun Kong, Kyu-Sup Cho
Tissue engineering cell-based therapy using induced pluripotent stem cells and adipose-derived stem cells (ASCs) may be promising tools for therapeutic applications in tissue engineering because of their abundance, relatively easy harvesting, and high proliferation potential. The purpose of this study was to investigate whether ASCs can promote the auricular cartilage regeneration in the rabbit. In order to assess their differentiation ability, ASCs were injected into the midportion of a surgically created auricular cartilage defect in the rabbit...
2018: Mediators of Inflammation
Muting Yan, Chen Xu, Yumei Huang, Huayue Nie, Jun Wang
The Three Gorges Project significantly impacted water quality and ecological balance in this area. The special engineered aquatic environment could be an important reservoir for antibiotic resistance genes (ARGs). Fifteen ARGs corresponding to three groups of antibiotics (tetracyclines, sulfonamides and quinolones) were determined in surface water, soil and sediment in this study. Total concentrations of antibiotics ranged from 21.55 to 536.86ng/L, 3.69 to 438.76ng/g, 15.78 to 213.84ng/g in water, soil and sediment, respectively...
August 1, 2018: Science of the Total Environment
Xin Qi, Hui Wang, Yadong Zhang, Libin Pang, Wei Xiao, Weitao Jia, Shichang Zhao, Deping Wang, Wenhai Huang, Qiugen Wang
Background: In the field of tissue engineering, there is currently increasing interest in new biomedical materials with high osteogenic ability and comparable mechanical function to repair bone defects. Three-dimensional (3-D) bioactive borosilicate glass (BG) scaffolds exhibit uniform interconnected macro-pores, high porosity and high compressive strength. In this study, we fabricated 3-D BG scaffolds by the 3D printing technique, then coated the surface of the 3-D BG scaffolds with mesoporous bioactive glass (MBG) (BG-MBG scaffold)...
2018: International Journal of Biological Sciences
Ling Ouyang, Yi Deng, Lei Yang, Xiuyuan Shi, Taosheng Dong, Youyi Tai, Weizhong Yang, Zhi-Gang Chen
Due to its similar elastic modulus of human bones, polyetheretherketone (PEEK) has been considered as an excellent cytocompatible material. However, the bioinertness, poor osteoconduction, and weak antibacterial activity of PEEK limit its wide applications in clinics. In this study, a facile strategy is developed to prepare graphene oxide (GO) modified sulfonated polyetheretherketone (SPEEK) (GO-SPEEK) through a simple dip-coating method. After detailed characterization, it is found that the GO closely deposits on the surface of PEEK, which is attributed to the π-π stacking interaction between PEEK and GO...
May 2, 2018: Macromolecular Bioscience
Mohammad M Pourseif, Gholamali Moghaddam, Hossein Daghighkia, Ahmad Nematollahi, Yadollah Omidi
Introduction: In this study, we targeted the worm stage of Echinococcus granulosus to design a novel multi-epitope B- and helper T-cell based vaccine construct for immunization of dogs against this multi-host parasite. Methods: The vaccine was designed based on the local Eg14-3-3 antigen (Ag). DNA samples were extracted from the protoscoleces of the infected sheep's liver, and then subjected to the polymerase chain reaction (PCR) with 14-3-3 specific forward and reverse primers. For the vaccine designing, several in silico steps were undertaken...
2018: BioImpacts: BI
Man Wang, Guan-Nan Liu, Hong Liu, Lu Zhang, Bing-Zhi Li, Xia Li, Duo Liu, Ying-Jin Yuan
Background: Evolution of complex phenotypes in cells requires simultaneously tuning expression of large amounts of genes, which can be achieved by reprograming global transcription. Lipophilicity is an important complex trait in oleaginous yeast Yarrowia lipolytica . It is necessary to explore the changes of which genes' expression levels will tune cellular lipophilic properties via the strategy of global transcription engineering. Results: We achieved a strategy of global transcription engineering in Y...
2018: Biotechnology for Biofuels
Joo Hee Choi, Hayan Jeon, Jeong Eun Song, Joaquim Miguel Oliveira, Rui Luis Reis, Gilson Khang
Cornea endothelial cells (CEnCs) tissue engineering is a great challenge to repair diseased or damaged CEnCs and require an appropriate biomaterial to support cell proliferation and differentiation. Biomaterials for CEnCs tissue engineering require biocompatibility, tunable biodegradability, transparency, and suitable mechanical properties. Silk fibroin-based film (SF) is known to meet these factors, but construction of functionalized graft for bioengineering of cornea is still a challenge. Herein, lysophosphatidic acid (LPA) is used to maintain and increase the specific function of CEnCs...
April 30, 2018: Nanomaterials
Zhicong Wang, Kaihua Li, Huijun Sun, Ji Wang, Zhuodong Fu, Mozhen Liu
Icariin, a traditional Chinese medicine, has previously been demonstrated to promote chondrogenesis of bone marrow mesenchymal stem cells (BMSCs) in traditional 2D cell culture. The present study investigated whether icariin has the potential to promote stable chondrogenic differentiation of BMSCs without hypertrophy in a 3D microenvironment. BMSCs were cultivated in a self‑assembling peptide nanofiber hydrogel scaffold in chondrogenic medium for 3 weeks. Icariin was added to the medium throughout the culture period at concentrations of 1x10‑6 M...
April 23, 2018: Molecular Medicine Reports
Caiyun Ma, Tengfei Lu, Hebao Wen, Yanjie Zheng, Xiao Han, Xongda Ji, Weijun Guan
Cartilage stem/progenitor cells (CSPCs) are a novel stem cell population and function as promising therapeutic candidates for cell‑based cartilage repair. Until now, numerous existing research materials have been obtained from humans, horses, cows and other mammals, but rarely from sheep. In the present study, CSPCs with potential applications in repairing tissue damage and cell‑based therapy were isolated from 45‑day‑old Small‑tailed Han Sheep embryos, and examined at the cellular and molecular level...
April 17, 2018: International Journal of Molecular Medicine
Hui Mei, Jen-Yu Liao, Randi M Jimenez, Yajun Wang, Saikat Bala, Cailen McCloskey, Christopher Switzer, John C Chaput
In vitro selection experiments carried out on artificial genetic polymers require robust and faithful methods for copying genetic information back and forth between DNA and xeno-nucleic acids (XNA). Previously, we have shown that Kod-RI, an engineered polymerase developed to transcribe DNA templates into threose nucleic acid (TNA), can function with high fidelity in the absence of manganese ions. However, the transcriptional efficiency of this enzyme diminishes greatly when individual templates are replaced with libraries of DNA sequences, indicating that manganese ions are still required for in vitro selection...
April 18, 2018: Journal of the American Chemical Society
Renze Shen, Weihong Xu, Yanxiang Xue, Luyuan Chen, Haicheng Ye, Enyi Zhong, Zhanchao Ye, Jie Gao, Yurong Yan
OBJECTIVE: In this study, nanofibrous scaffolds base on pure polylactic acid (PLA) and chitosan/PLA blends were fabricated by emulsion eletrospinning. By modulating their mechanical and biological properties, cell-compatible and biodegradable scaffolds were developed for periodontal bone regeneration. METHODS: Pure PLA and different weight ratios of chitosan nano-particle/PLA nano-fibers were fabricated by emulsion eletrospinning. Scanning electron microscope (SEM) was performed to observe the morphology of nano-fibers...
April 16, 2018: Artificial Cells, Nanomedicine, and Biotechnology
Pruthvi Raj Bejugam, Shailza Singh
Post-transcriptional gene regulation is a vital process to regulate expression of the key genes in the eukaryotic cell. Such processes are essential for pathogens which reside inside the host cell. One such pathogen is Leishmania major, which causes cutaneous leishmaniasis. The parasite lives inside the macrophages of mammalian host (mostly human). Inside the macrophage, Leishmania genes show complex host-pathogen interaction regulating a plethora of gene expression. Till date, most of the studies have shown this kind of regulation with respect to the host macrophages...
April 12, 2018: International Journal of Biological Macromolecules
Ying Lu, Zhuojun Wang, Lu Chen, Jia Wang, Shulin Li, Caixia Liu, Dong Sun
Amniotic fluid is an alternative source of stem cells, and human amniotic fluid-derived stem cells (AFSCs) obtained from a small amount of amniotic fluid collected during the second trimester represent a novel source for use in regenerative medicine. These AFSCs are characterized by lower diversity, a higher proliferation rate and a wider differentiation capability than adult mesenchymal stem cells (MSCs). AFSCs are selected based on the cell surface marker c-kit receptor (CD117) using immunomagnetic sorting...
March 15, 2018: Stem Cells and Development
X T Wang, N Q Rao, T Jz Fang, Y M Zhao, L H Ge
OBJECTIVE: Stem cells from human exfoliated teeth (SHED) were sorted by magnetically activated cell sorting (MACS) technique to obtain the CD146 positive and negative cell subpopulation. Then the biological characteristics of these subpopulations were compared to explore their specific application potential in tissue engineering. METHODS: In this study, freshly extracted deciduous teeth without any caries or dental pulp disease were obtained. SHED was isolated using enzyme digestion method and then sorted by MACS, CD146 positive cells and CD146 negative cells were obtained after cell sorting...
April 18, 2018: Beijing da Xue Xue Bao. Yi Xue Ban, Journal of Peking University. Health Sciences
Haihui Liu, Mingtai Chen, Lulu Liu, Saisai Ren, Panpan Cheng, Hao Zhang
Accumulating evidence indicates that mesenchymal stem cells (MSCs) have been widely used in tissue engineering and regenerative medicine due to their multilineage differentiation potentials. Recent studies show that germ-like cells can also be derived from stem cells, such as human umbilical cord MSCs and human bone marrow MSCs in vitro. However, whether human adipose-derived MSCs (hAD-MSCs) can be induced into germ-like cells has never been reported. In this study, we isolated hAD-MSCs and confirmed that their characteristics were in accordance with that of MSCs established before...
April 2018: Cellular Reprogramming
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