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Polymerase engineering

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https://www.readbyqxmd.com/read/28420224/silk-fibroin-alginate-hydroxyapatite-composite-particles-in-bone-tissue-engineering-applications-in-vivo
#1
You-Young Jo, Seong-Gon Kim, Kwang-Jun Kwon, HaeYong Kweon, Weon-Sik Chae, Won-Geun Yang, Eun-Young Lee, Hyun Seok
The aim of this study was to evaluate the in vivo bone regeneration capability of alginate (AL), AL/hydroxyapatite (HA), and AL/HA/silk fibroin (SF) composites. Forty Sprague Dawley rats were used for the animal experiments. Central calvarial bone (diameter: 8.0 mm) defects were grafted with AL, AL/HA, or AL/HA/SF. New bone formation was evaluated by histomorphometric analysis. To demonstrate the immunocompatibility of each group, the level of tumor necrosis factor (TNF)-α expression was studied by immunohistochemistry (IHC) and quantitative reverse transcription polymerase chain reaction (qRT-PCR) at eight weeks post implantation...
April 18, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28419944/electrochemical-synthesis-of-three-dimensional-porous-reduced-graphene-oxide-film-preparation-and-in-vitro-osteogenic-activity-evaluation
#2
Zizhu Tian, Lixun Huang, Xibo Pei, Junyu Chen, Tong Wang, Tao Yang, Han Qin, Lei Sui, Jian Wang
In this study, three-dimensional reduced graphene oxide (3D-rGO) porous films were fabricated using a two-step electrochemical method, including an electrochemical deposition process for the self-assembly of GO and an electrochemical bubbling-based transfer. The morphology, physical properties, and phase composition of the 3D-rGO films were characterized, and the cellular bioactivities were evaluated using pre-osteoblasts (MC3T3-E1 cells). The attachment, proliferation and differentiation of the MC3T3-E1 cells on the 3D-rGO films was analyzed by scanning electron microscopy (SEM), Cell Counting Kit-8 (CCK-8) assays and live/dead cell staining, and alkaline phosphatase (ALP) activity assays, respectively...
April 12, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28409292/effects-of-the-donor-age-on-proliferation-senescence-and-osteogenic-capacity-of-human-urine-derived-stem-cells
#3
Peng Gao, Peilin Han, Dapeng Jiang, Shulong Yang, Qingbo Cui, Zhaozhu Li
To study the effects of the donor age on the application potential of human urine-derived stem cells (hUSCs) in bone tissue engineering, by comparing proliferation, senescence and osteogenic differentiation of hUSCs originated from volunteers with different ages. The urine samples were collected from 19 healthy volunteers (6 cases from children group aged from 5 to 14, 5 cases from middle-aged group aged from 30 to 40, and 8 cases from the elder group aged from 65 to 75), and hUSCs were isolated and cultured...
April 13, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28383245/exploring-the-chemistry-of-genetic-information-storage-and-propagation-through-polymerase-engineering
#4
Gillian Houlihan, Sebastian Arangundy-Franklin, Philipp Holliger
Nucleic acids are a distinct form of sequence-defined biopolymer. What sets them apart from other biopolymers such as polypeptides or polysaccharides is their unique capacity to encode, store, and propagate genetic information (molecular heredity). In nature, just two closely related nucleic acids, DNA and RNA, function as repositories and carriers of genetic information. They therefore are the molecular embodiment of biological information. This naturally leads to questions regarding the degree of variation from this seemingly ideal "Goldilocks" chemistry that would still be compatible with the fundamental property of molecular heredity...
April 6, 2017: Accounts of Chemical Research
https://www.readbyqxmd.com/read/28381166/downregulation-of-microrna-181d-had-suppressive-effect-on-pancreatic-cancer-development-through-inverse-regulation-of-knain2
#5
Guopeng Zhang, Dongbo Liu, Guoxian Long, Lei Shi, Hong Qiu, Guangyuan Hu, Guoqing Hu, Shunfang Liu
We explored the expression and function of miR-181d (microRNA-181d) in human pancreatic cancer. Quantitative real-time polymerase chain reaction was used to probe miR-181d expression in both pancreatic cancer cell lines and human pancreatic carcinoma. Pancreatic cancer cell lines, PANC-1 and AsPC-1 cells, were engineered to stably downregulate endogenous miR-181d through lentiviral transduction. The mechanistic effects of miR-181d downregulation on pancreatic cancer development were tested by proliferation, migration, fluorouracil chemosensitivity assays in vitro, and explant assay in vivo...
April 2017: Tumour Biology: the Journal of the International Society for Oncodevelopmental Biology and Medicine
https://www.readbyqxmd.com/read/28369677/direct-isolation-of-seamless-mutant-bacterial-artificial-chromosomes
#6
George T Lyozin, Yasuhiro Kosaka, Gourab Bhattacharje, H Joseph Yost, Luca Brunelli
Seamless (i.e., without unwanted DNA sequences) mutant bacterial artificial chromosomes (BACs) generated via recombination-mediated genetic engineering (recombineering) are better suited to study gene function compared to complementary DNA (cDNA) because they contain only the specific mutation and provide all the regulatory sequences required for in vivo gene expression. However, precisely mutated BACs are typically rare (∼1:1,000 to 1:100,000), making their isolation quite challenging. Although these BACs have been classically isolated by linking the mutation to additional genes, i...
April 3, 2017: Current Protocols in Molecular Biology
https://www.readbyqxmd.com/read/28367415/effects-of-electromagnetic-stimulation-on-gene-expression-of-mesenchymal-stem-cells-and-repair-of-bone-lesions
#7
Maryam Jazayeri, Mohammad Ali Shokrgozar, Nooshin Haghighipour, Bahram Bolouri, Fereshteh Mirahmadi, Mehdi Farokhi
OBJECTIVE: Most people experience bone damage and bone disorders during their lifetimes. The use of autografts is a suitable way for injury recovery and healing. Mesenchymal stem cells (MSCs) are key players in tissue engineering and regenerative medicine. Their proliferation potential and multipotent differentiation ability enable MSCs to be considered as appropriate cells for therapy and clinical applications. Differentiation of stem cells depends on their microenvironment and biophysical stimulations...
April 2017: Cell Journal
https://www.readbyqxmd.com/read/28352329/matrigel-scaffold-combined-with-ad-hbmp7-transfected-chondrocytes-improves-the-repair-of-rabbit-cartilage-defect
#8
Xiaopeng Xia, Jing Li, Bo Xia, Huilin Yang, Dongmei Zhang, Bin Zhou, Jie Zhang, Man Zhou, Fan Liu
The aim of this study was to explore an effective method for the repair of cartilage defects using chitosan/glycerophosphate (C/GP) gel- and Matrigel-engineered human bone morphogenetic protein 7 (hBMP7)-expressing chondrocytes. Rabbit chondrocytes were obtained, cultured in vitro and transfected with an adenovirus containing hBMP7 and green fluorescent protein (Ad-hBMP7-GFP). The expression of hBMP7 in the transfected cells was tested by reverse transcription-polymerase chain reaction (RT-PCR) and western blotting...
February 2017: Experimental and Therapeutic Medicine
https://www.readbyqxmd.com/read/28350666/long-term-comparison-between-human-normal-conchal-and-microtia-chondrocytes-regenerated-by-tissue-engineering-on-nanofiber-polyglycolic-acid-scaffolds
#9
Hitomi Nakao, Robin D Jacquet, Mark Shasti, Noritaka Isogai, Ananth S Murthy, William J Landis
BACKGROUND: Previous regeneration studies of auricle-shaped cartilage by tissue engineering leave unresolved whether the chondrocyte phenotype from human auricular chondrocytes seeded onto polymeric scaffolds is retained over the long term and whether microtia remnants may be a viable cell source for auricular reconstruction. METHODS: Chondrocytes were isolated from human ears, either normal conchal ear or microtia cartilage remnants, expanded in vitro, and seeded onto nanoscale-diameter polyglycolic acid sheets...
April 2017: Plastic and Reconstructive Surgery
https://www.readbyqxmd.com/read/28349897/3d-printing-scaffold-coupled-with-low-level-light-therapy-for-neural-tissue-regeneration
#10
Wei Zhu, Jonathan K George, Volker J Sorger, Lijie Grace Zhang
3D printing has shown promise for neural regeneration by providing customized nerve scaffolds to structurally support and bridge the defect gap as well as deliver cells or various bioactive substances. Low-level light therapy (LLLT) exhibits positive effects on rehabiliation of degenerative nerves and neural disorders. With this in mind, we postulate that 3D printed neural scaffold coupling with LLLT will generate a new strategy to repair neural degeneration. To achieve this goal, we applied red laser light to stimualte neural stem cells on 3D printed scaffolds and investigated the subsequent cell response with respect to cell proliferation and differentiation...
April 12, 2017: Biofabrication
https://www.readbyqxmd.com/read/28349101/data-of-self-made-taq-dna-polymerase-prepared-for-screening-purposes
#11
E V Konovalova, A A Schulga, T I Lukyanova, E J Woo, S M Deyev
DNA analysis is a key procedure in genetic engineering. Nowadays the analysis is often done by PCR with Taq DNA polymerase. Although the last enzyme price is quite low, demand for numerous analyses results in much money expenditure which are not affordable for many laboratories. In a meanwhile, many screening tasks do not require the highly purified enzyme. Taking into account the enzyme unique properties it makes possible to marginally simplify its production without resorting to costly or lengthy techniques such as column chromatography and/or dialysis...
April 2017: Data in Brief
https://www.readbyqxmd.com/read/28345449/biocompatibility-of-pcl-plga-bcp-porous-scaffold-for-bone-tissue-engineering-applications
#12
Nguyen Thi Hiep, Huynh Chan Khon, Nguyen Dai Hai, Lee Byong-Taek, Vo Van Toi, Le Thanh Hung
In this study, biomimic porous polycaprolactone/poly (lactide-co-glycolide) loading biphasic tricalcium phosphate (PCL/PLGA-BCP) scaffolds were fabricated successfully by solvent evaporation method. The distribution of biphasic tricalcium phosphate (BCP) in polycaprolactone/poly (lactide-co-glycolide) (PCL/PLGA) scaffold was confirmed by micro-computed tomography (micro-CT) scanning, scanning electron microscope (SEM) observation and Energy-dispersive X-ray Spectroscopy (EDS) analysis. The hydrophilicity of the scaffolds was confirmed by contact angle measurement...
June 2017: Journal of Biomaterials Science. Polymer Edition
https://www.readbyqxmd.com/read/28303150/development-of-synebrick-vectors-as-a-synthetic-biology-platform-for-gene-expression-in-synechococcus-elongatus-pcc-7942
#13
Wook Jin Kim, Sun-Mi Lee, Youngsoon Um, Sang Jun Sim, Han Min Woo
Cyanobacteria are oxygenic photosynthetic prokaryotes that are able to assimilate CO2 using solar energy and water. Metabolic engineering of cyanobacteria has suggested the possibility of direct CO2 conversion to value-added chemicals. However, engineering of cyanobacteria has been limited due to the lack of various genetic tools for expression and control of multiple genes to reconstruct metabolic pathways for biochemicals from CO2. Thus, we developed SyneBrick vectors as a synthetic biology platform for gene expression in Synechococcus elongatus PCC 7942 as a model cyanobacterium...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28292602/investigation-of-human-dental-pulp-cells-on-a-potential-injectable-poly-lactic-co-glycolic-acid-microsphere-scaffold
#14
Huiru Zou, Guanhua Wang, Fang Song, Xudong Shi
INTRODUCTION: Poly(lactic-co-glycolic acid) (PLGA) has been extensively explored in the tissue engineering field with good biocompatibility and biodegradability. PLGA microspheres' injectable potency makes it highly desirable in dentin-pulp complex regeneration. Therefore, we investigated the cell adhesion, proliferation, odontogenic differentiation, and matrix mineralization of human dental pulp cells (HDPCs) on a PLGA microsphere scaffold. We hypothesized that this scaffold might be suitable for dentin-pulp complex regeneration...
March 11, 2017: Journal of Endodontics
https://www.readbyqxmd.com/read/28254397/high-level-expression-of-two-thermophilic-%C3%AE-mannanases-in-yarrowialipolytica
#15
Wang YaPing, Rao Ben, Zhang Ling, Ma Lixin
Two thermophilic β-mannanases (ManA and ManB)were successfully expressed in Yarrowialipolytica using vector pINA1296I. The sequences of manA from Aspergillus niger CBS 513.88 and manB from Bacillus subtilis BCC41051 were optimized based on codon-usage bias in Y.lipolytica and synthesized by overlapping polymerase chain reaction (PCR). We utilized the pINA1296I vector, which allows inserting and expression of multiple copies of an expression cassette, to engineer recombinant strains containing multiple copies of manA or manB...
February 22, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28247198/development-and-application-of-a-t7-rna-polymerase-dependent-expression-system-for-antibiotic-production-improvement-in-streptomyces
#16
Junhong Wei, Jinjin Tian, Guoqing Pan, Jie Xie, Jialing Bao, Zeyang Zhou
OBJECTIVE: To develop a reliable and easy to use expression system for antibiotic production improvement of Streptomyces. RESULTS: A two-compound T7 RNA polymerase-dependent gene expression system was developed to fulfill this demand. In this system, the T7 RNA polymerase coding sequence was optimized based on the codon usage of Streptomyces coelicolor. To evaluate the functionality of this system, we constructed an activator gene overexpression strain for enhancement of actinorhodin production...
February 28, 2017: Biotechnology Letters
https://www.readbyqxmd.com/read/28245548/high-prevalence-and-genetic-polymorphisms-of-legionella-in-natural-and-man-made-aquatic-environments-in-wenzhou-china
#17
Leyi Zhang, Yi Li, Xin Wang, Zhihui Shangguan, Haijian Zhou, Yuejin Wu, Lianghuai Wang, Hongyu Ren, Yun Hu, Meifen Lin, Tian Qin
Natural and engineered water systems are the main sources of Legionnaires' disease. It is essential from a public health perspective to survey water environments for the existence of Legionella. To analyze the main serogroups, genotypes and pathogenicity of the pathogen, a stratified sampling method was adopted to collect water samples randomly from shower water, cooling tower water, and local public hot springs in Wenzhou, China. Suspected strains were isolated from concentrated water samples. Serum agglutination assay and real-time PCR (Polymerase chain reaction) were used to identify L...
February 24, 2017: International Journal of Environmental Research and Public Health
https://www.readbyqxmd.com/read/28230059/synergistic-effects-of-dimethyloxallyl-glycine-and-recombinant-human-bone-morphogenetic-protein-2-on-repair-of-critical-sized-bone-defects-in-rats
#18
Xin Qi, Yang Liu, Zhen-Yu Ding, Jia-Qing Cao, Jing-Huan Huang, Jie-Yuan Zhang, Wei-Tao Jia, Jing Wang, Chang-Sheng Liu, Xiao-Lin Li
In bone remodeling, osteogenesis is closely coupled to angiogenesis. Bone tissue engineering using multifunctional bioactive materials is a promising technique which has the ability to simultaneously stimulate osteogenesis and angiogenesis for repair of bone defects. We developed mesoporous bioactive glass (MBG)-doped poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) composite scaffolds as delivery vehicle. Two bioactive molecules, dimethyloxalylglycine (DMOG), a small-molecule angiogenic drug, and recombinant human bone morphogenetic protein-2 (rhBMP-2), an osteoinductive growth factor, were co-incorporated into the scaffold...
February 23, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28218004/synthesis-and-characterization-of-some-abundant-nanoparticles-their-antimicrobial-and-enzyme-inhibition-activity
#19
Shams T Khan, Ajmaluddin Malik, Rizwan Wahab, Omar H Abd-Elkader, Maqusood Ahamed, Javed Ahmad, Javed Musarrat, Maqsood A Siddiqui, Abdulaziz A Al-Khedhairy
Although the antimicrobial activity of the engineered nanoparticles (NPs) is well known, the biochemical mechanisms underlying this activity are not clearly understood. Therefore, four NPs with the highest global production, namely SiO2, TiO2, ZnO, and Ag, were synthesized and characterized. The synthesized SiO2, TiO2, ZnO, and Ag NPs exhibit an average size of 11.12, 13.4, 35, and 50 nm, respectively. The antimicrobial activity of the synthesized NPs against bacteria and fungi were also determined. NPs-mediated inhibition of two very important enzymes, namely urease and DNA polymerase, is also reported...
February 20, 2017: Acta Microbiologica et Immunologica Hungarica
https://www.readbyqxmd.com/read/28208291/growth-based-bacterial-viability-assay-for-interference-free-and-high-throughput-toxicity-screening-of-nanomaterials
#20
Tian A Qiu, Thu Ha Thi Nguyen, Natalie V Hudson-Smith, Peter L Clement, Dona-Carla Forester, Hilena Frew, Mimi N Hang, Catherine J Murphy, Robert J Hamers, Z Vivian Feng, Christy L Haynes
Current high-throughput approaches evaluating toxicity of chemical agents toward bacteria typically rely on optical assays, such as luminescence and absorbance, to probe the viability of the bacteria. However, when applied to toxicity induced by nanomaterials, scattering and absorbance from the nanomaterials act as interferences that complicate quantitative analysis. Herein, we describe a bacterial viability assay that is free of optical interference from nanomaterials and can be performed in a high-throughput format on 96-well plates...
February 7, 2017: Analytical Chemistry
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