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https://www.readbyqxmd.com/read/28922845/pot1-mediated-%C3%AE-integration-strategy-for-high-copy-stable-expression-of-heterologous-proteins-in-saccharomyces-cerevisiae
#1
Xiaofei Song, Quanli Liu, Jiwei Mao, Yuzhen Wu, Yuanzi Li, Kai Gao, Xiuming Zhang, Yanling Bai, Haijin Xu, Mingqiang Qiao
In biotechnological industry, increased expression cassette stability and copy number serve as important means of maintaining consistently high production levels of heterologous proteins in Saccharomyces cerevisiae. In this study, we combined δ sequences for site-specific integration with TPI1 gene from Schizosaccharomyces pombe (POT1) as a selection marker to realize high-copy integration and stable expression of heterologous proteins in S. cerevisiae. With the newly developed POT1 platform, a 32-copy integration of enhanced green fluorescent protein (EGFP) expression cassette was obtained in a single round and was stably maintained after 100 generations of growth in a rich complex medium...
September 1, 2017: FEMS Yeast Research
https://www.readbyqxmd.com/read/28922717/magnetic-nanoparticle-loaded-alginate-beads-for-local-micro-actuation-of-in-vitro-tissue-constructs
#2
Awatef M Alshehri, Otto C Wilson, Bishnu Dahal, John Philip, Xiaolong Luo, Christopher B Raub
Magnetic nanoparticles (MNPs) self-align and transduce magnetic force, two properties which lead to promising applications in cell and tissue engineering. However, the toxicity of MNPs to cells which uptake them is a major impediment to applications in engineered tissue constructs. To address this problem, MNPs were embedded in millimeter-scale alginate beads, coated with glutaraldehyde cross-linked chitosan, and loaded in acellular and MDA-MB-231 cancer cell-seeded collagen hydrogels, providing local micro-actuation under an external magnetic field...
September 4, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28922664/patchy-particles-made-by-colloidal-fusion
#3
Zhe Gong, Theodore Hueckel, Gi-Ra Yi, Stefano Sacanna
Patches on the surfaces of colloidal particles provide directional information that enables the self-assembly of the particles into higher-order structures. Although computational tools can make quantitative predictions and can generate design rules that link the patch motif of a particle to its internal microstructure and to the emergent properties of the self-assembled materials, the experimental realization of model systems of particles with surface patches (or 'patchy' particles) remains a challenge. Synthetic patchy colloidal particles are often poor geometric approximations of the digital building blocks used in simulations and can only rarely be manufactured in sufficiently high yields to be routinely used as experimental model systems...
September 18, 2017: Nature
https://www.readbyqxmd.com/read/28922651/evaluating-the-effect-of-smoking-cessation-treatment-on-a-complex-dynamical-system
#4
Korkut Bekiroglu, Michael A Russell, Constantino M Lagoa, Stephanie T Lanza, Megan E Piper
OBJECTIVE: To understand the dynamic relations among tobacco withdrawal symptoms to inform the development of effective smoking cessation treatments. Dynamical system models from control engineering are introduced and utilized to evaluate complex treatment effects. We demonstrate how dynamical models can be used to examine how distinct withdrawal-related processes are related over time and how treatment influences these relations. METHOD: Intensive longitudinal data from a randomized placebo-controlled smoking cessation trial (N=1504) are used to estimate a dynamical model of withdrawal-related processes including momentary craving, negative affect, quitting self-efficacy, and cessation fatigue for each of six treatment conditions (nicotine patch, nicotine lozenge, bupropion, patch + lozenge, bupropion + lozenge, and placebo)...
August 25, 2017: Drug and Alcohol Dependence
https://www.readbyqxmd.com/read/28922528/mass-production-and-pore-size-control-of-holey-carbon-microcages
#5
Lei Zhang, Xiaoxiao Liu, Yuhai Dou, Binwei Zhang, Huiling Yang, Shixue Dou, Huakun Liu, Yunhui Huang, Xianluo Hu
Architectural control of porous solids, such as porous carbon cages, has received considerable attention for versatile applications because of their attractive ability to interact with liquids and gases not only at the surface, but throughout the bulk. Here we report a scalable, facile spray-pyrolysis route to synthesize holey carbon microcages with mosquito-net-like shells. Using the surfaces of water droplets as the growth templates, styrene-butadiene rubber macromolecules are controllably cross-linked, and size-tunable holes on the carbon shells are generated...
September 18, 2017: Angewandte Chemie
https://www.readbyqxmd.com/read/28922409/purification-and-characterisation-of-the-yeast-plasma-membrane-atp-binding-cassette-transporter-pdr11p
#6
Katrine Rude Laub, Magdalena Marek, Lyubomir Dimitrov Stanchev, Sara Abad Herrera, Tamara Kanashova, Adèle Bourmaud, Gunnar Dittmar, Thomas Günther Pomorski
The ATP binding cassette (ABC) transporters Pdr11p and its paralog Aus1p are expressed under anaerobic growth conditions at the plasma membrane of the yeast Saccharomyces cerevisiae and are required for sterol uptake. However, the precise mechanism by which these ABC transporters facilitate sterol movement is unknown. In this study, an overexpression and purification procedure was developed with the aim to characterise the Pdr11p transporter. Engineering of Pdr11p variants fused at the C terminus with green fluorescent protein (Pdr11p-GFP) and containing a FLAG tag at the N terminus facilitated expression analysis and one-step purification, respectively...
2017: PloS One
https://www.readbyqxmd.com/read/28922403/the-declining-interest-in-an-academic-career
#7
Michael Roach, Henry Sauermann
There is increasing evidence that science & engineering PhD students lose interest in an academic career over the course of graduate training. It is not clear, however, whether this decline reflects students being discouraged from pursuing an academic career by the challenges of obtaining a faculty job or whether it reflects more fundamental changes in students' career goals for reasons other than the academic labor market. We examine this question using a longitudinal survey that follows a cohort of PhD students from 39 U...
2017: PloS One
https://www.readbyqxmd.com/read/28922354/reconstructing-promoter-activity-from-lux-bioluminescent-reporters
#8
Mudassar Iqbal, Neil Doherty, Anna M L Page, Saara N A Qazi, Ishan Ajmera, Peter A Lund, Theodore Kypraios, David J Scott, Philip J Hill, Dov J Stekel
The bacterial Lux system is used as a gene expression reporter. It is fast, sensitive and non-destructive, enabling high frequency measurements. Originally developed for bacterial cells, it has also been adapted for eukaryotic cells, and can be used for whole cell biosensors, or in real time with live animals without the need for euthanasia. However, correct interpretation of bioluminescent data is limited: the bioluminescence is different from gene expression because of nonlinear molecular and enzyme dynamics of the Lux system...
September 18, 2017: PLoS Computational Biology
https://www.readbyqxmd.com/read/28922184/subjective-measures-of-work-related-fatigue-in-automobile-factory-employees
#9
Fu Guo, Tianbo Wang, Zuojiang Ning
BACKGROUND: Work-related fatigue is common among automobile factory employees. OBJECTIVES: The purpose of this study was to assess fatigue of employees at a Chinese automobile factory. METHODS: 238 employees (119 engineers and 119 workers) participated in this study. The following questionnaires were completed: demographic survey questionnaire, working condition questionnaire (WCQ), functional assessment of chronic illness therapy-fatigue (FACIT-F), subscales of multidimensional fatigue inventory (MFI), and Pittsburgh sleep quality index (PSQI)...
September 14, 2017: Work: a Journal of Prevention, Assessment, and Rehabilitation
https://www.readbyqxmd.com/read/28921994/anisotropic-cracking-of-nanocrystal-superlattices
#10
Benjamin T Diroll, Xuedan Ma, Yaoting Wu, Christopher B Murray
The synthesis colloidal nanocrystals in non-polar organic solvents has led to exceptional size- and shape-control, enabling the formation of nanocrystal superlattices isostructural to atomic lattices built with nanocrystals rather than atoms. The long aliphatic ligands (e.g. oleic acid) used to achieve this control separate nanocrystals too far in the solid-state for most charge-transporting devices. Solid-state ligand exchange, which brings particles closer together and enhances conductivity, necessitates large changes in the total volume of the solid (compressive stress), which leads to film cracking...
September 18, 2017: Nano Letters
https://www.readbyqxmd.com/read/28921949/effects-of-high-temperature-and-thermal-cycling-on-the-performance-of-perovskite-solar-cells-acceleration-of-charge-recombination-and-deterioration-of-charge-extraction
#11
Arif D Sheikh, Rahim Munir, Md Azimul Haque, Ashok Bera, Weijin Hu, Parvez Shaikh, Aram Amassian, Tom Wu
In this work, we investigated the effects of high operating temperature and thermal cycling on the photovoltaic performance of perovskite solar cells (PSCs) with a typical mesostructured (m)-TiO2-CH3NH3PbI3-xClx-spiro-OMeTAD architecture. After carrying out temperature-dependent grazing incidence wide-angle X-ray scattering (GIWAXS), in-situ X-ray diffraction (XRD) and optical absorption experiments, thermal durability of PSCs was tested by subjecting the devices to repetitive heating to 70 °C and cooling to room temperature (20 °C)...
September 18, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28921946/biomimetic-magnetosomes-as-versatile-artificial-antigen-presenting-cells-to-potentiate-t-cell-based-anticancer-therapy
#12
Qianmei Zhang, Wei Wei, Peiling Wang, Liping Zuo, Feng Li, Jin Xu, Xiaobo Xi, Xiaoyong Gao, Guanghui Ma, Hai-Yan Xie
Adoptive T-cell transfer for cancer therapy relies on both effective ex vivo T cell expansion and in vivo targeting performance. One promising but challenging method for accomplishing this purpose is to construct multifunctional artificial antigen-presenting cells (aAPCs). We herein developed biomimetic magnetosomes as versatile aAPCs, wherein magnetic nanoclusters were coated with azide-engineered leucocyte membranes and then decorated with T-cell stimuli through copper-free click chemistry. These novel nano aAPCs not only exhibited high performance for antigen-specific cytotoxic T cell (CTL) expansion and stimulation but also visually and effectively guided reinfused CTLs to tumor tissues through magnetic resonance imaging and magnetic control...
September 18, 2017: ACS Nano
https://www.readbyqxmd.com/read/28921830/trends-application-and-future-prospectives-of-microbial-carbonic-anhydrase-mediated-carbonation-process-for-ccus
#13
REVIEW
Chintan Bhagat, Pravin Dudhagara, Shantilal Tank
Growing industrialisation and desire for better economy of countries accelerated the emission of greenhouse gasses, more than the buffering capacity of earth atmosphere. Among, various Green House Gasses (GHGs), CO2 occupies the first position in anthroposphere and has detrimental effects on the ecosystem. For decarbonization, several non-biological methods of carbon capture, utilisation, and storage (CCUS) are in use from last few decades, but they are suffering from narrow applicability. Recently, CO2 emission and its disposal related problem encourage for implementation of bioprocessing to achieve zero waste economy for a sustainable environment...
September 18, 2017: Journal of Applied Microbiology
https://www.readbyqxmd.com/read/28921822/construction-of-human-induced-pluripotent-stem-cell-derived-oriented-bone-matrix-microstructure-by-using-in-vitro-engineered-anisotropic-culture-model
#14
Ryosuke Ozasa, Aira Matsugaki, Yoshihiro Isobe, Taro Saku, Hui-Suk Yun, Takayoshi Nakano
Bone tissue has anisotropic microstructure based on collagen/biological apatite orientation, which plays essential roles in the mechanical and biological functions of bone. However, obtaining an appropriate anisotropic microstructure during the bone regeneration process remains a great challenging. A powerful strategy for the control of both differentiation and structural development of newly-formed bone is required in bone tissue engineering, in order to realize functional bone tissue regeneration. In this study, we developed a novel anisotropic culture model by combining human induced pluripotent stem cells (hiPSCs) and artificially-controlled oriented collagen scaffold...
September 16, 2017: Journal of Biomedical Materials Research. Part A
https://www.readbyqxmd.com/read/28921815/low-gluten-non-transgenic-wheat-engineered-with-crispr-cas9
#15
Susana Sánchez-León, Javier Gil-Humanes, Carmen V Ozuna, María J Giménez, Carolina Sousa, Daniel F Voytas, Francisco Barro
Celiac disease is an autoimmune disorder triggered in genetically predisposed individuals by the ingestion of gluten proteins from wheat, barley, and rye. The -gliadin gene family of wheat contains four highly stimulatory peptides, of which the 33-mer is the main immunodominant peptide in celiac patients. We designed two sgRNAs to target a conserved region adjacent to the coding sequence for the 33-mer in the -gliadin genes. Twenty-one mutant lines were generated, all showing strong reduction in -gliadins. Up to 35 different genes were mutated in one of the lines of the 45 different genes identified in the wild type, while immunoreactivity was reduced by 85%...
September 18, 2017: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/28921578/the-darknet-the-new-street-for-street-drugs
#16
J V Pergolizzi, J A LeQuang, R Taylor, R B Raffa
WHAT IS KNOWN AND OBJECTIVE: The "Darknet" ("dark web") has emerged as a means by which illegal drug buys and deliveries can be arranged with apparent anonymity and impunity. Healthcare providers should be aware of this growing source of illicit drugs. COMMENT: The "Darknet" refers to networks isolated from the Internet that cannot be accessed via conventional search engines. They require special software that is protected by special encryption. The initial legitimate use of a "Darknet" to conceal personal information against misuse or political reprisal is being exploited to conceal the identity of buyers and sellers in illegal drug transactions...
September 15, 2017: Journal of Clinical Pharmacy and Therapeutics
https://www.readbyqxmd.com/read/28921549/%C3%AE-flucs-brighter-photinus-pyralis-firefly-luciferases-identified-by-surveying-consecutive-single-amino-acid-deletion-mutations-in-a-thermostable-variant
#17
Lisa M Halliwell, Amit P Jathoul, Jack P Bate, Harley L Worthy, James C Anderson, Dafydd D Jones, James A H Murray
The bright bioluminescence catalysed by Photinus pyralis firefly luciferase (Fluc) enables a vast array of life science research such as bioimaging in live animals and sensitive in vitro diagnostics. The effectiveness of such applications is improved using engineered enzymes that to date have been constructed using amino acid substitutions. We describe ΔFlucs: consecutive single amino acid deletion mutants within 6 loop structures of the bright and thermostable x11 Fluc. Deletion mutations are a promising avenue to explore new sequence and functional space and isolate novel mutant phenotypes...
September 16, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28921540/imaging-stem-cell-distribution-growth-migration-and-differentiation-in-3-d-scaffolds-for-bone-tissue-engineering-using-mesoscopic-fluorescence-tomography
#18
Qinggong Tang, Charlotte Piard, Jonathan Lin, Kai Nan, Ting Guo, John Caccamese, John Fisher, Yu Chen
Regenerative medicine has emerged as an important discipline that aims to repair injury or replace damaged tissues or organs by introducing living cells or functioning tissues. Successful regenerative medicine strategies will likely depend upon a simultaneous optimization strategy for the design of biomaterials, cell-seeding methods, cell-biomaterial interactions and molecular signaling within the engineered tissues. It remains a challenge to image three-dimensional (3-D) structures and functions of the cell-seeded scaffold in mesoscopic scale (>2∼3 mm)...
September 16, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28921510/development-of-a-formaldehyde-biosensor-with-application-to-synthetic-methylotrophy
#19
Benjamin M Woolston, Timothy Roth, Ishwar Kohale, David Liu, Gregory Stephanopoulos
Formaldehyde is a prevalent environmental toxin and a key intermediate in single carbon metabolism. The ability to monitor formaldehyde concentration is therefore of interest for both environmental monitoring and for metabolic engineering of native and synthetic methylotrophs, but current methods suffer from low sensitivity, complex workflows, or require expensive analytical equipment. Here we develop a formaldehyde biosensor based on the FrmR repressor protein and cognate promoter of E. coli. Optimization of the native repressor binding site and regulatory architecture enabled detection at levels as low as 1 µM...
September 16, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28921443/polyester-based-nanoparticles-for-the-encapsulation-of-monoclonal-antibodies
#20
Flávia Sousa, Pedro Fonte, Andreia Cruz, Patrick J Kennedy, Inês Mendes Pinto, Bruno Sarmento
Aliphatic polyesters have been widely explored for biomedical applications (e.g., drug delivery systems, biomedical devices, and tissue engineering). Recently, polyesters have been used in nanoparticle formulations for the controlled release of monoclonal antibodies (mAbs) for the enhanced efficacy of antibody-based therapy. Polyester-based nanoparticles for mAb delivery provide decreased antibody dosage, increased antibody stability and protection and longer therapeutic action, ultimately translating to an increased therapeutic index...
2018: Methods in Molecular Biology
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