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David J Lea-Smith, Ravendran Vasudevan, Christopher J Howe
Cyanobacteria are ecologically important organisms and potential platforms for production of biofuels and useful industrial products. Genetic manipulation of cyanobacteria, especially model organisms such as Synechocystis sp. PCC6803 and Synechococcus sp. PCC7002, is a key tool for both basic and applied research. Generation of unmarked mutants, whereby chromosomal alterations are introduced into a strain via insertion of an antibiotic resistance cassette (a manipulatable fragment of DNA containing one or more genes), followed by subsequent removal of this cassette using a negative selectable marker, is a particularly powerful technique...
2016: Journal of Visualized Experiments: JoVE
Aayushi Uberoi, Satoshi Yoshida, Ian H Frazer, Henry C Pitot, Paul F Lambert
Human papillomaviruses are causally associated with 5% of human cancers. The recent discovery of a papillomavirus (MmuPV1) that infects laboratory mice provides unique opportunities to study the life cycle and pathogenesis of papillomaviruses in the context of a genetically manipulatable host organism. To date, MmuPV1-induced disease has been found largely to be restricted to severely immunodeficient strains of mice. In this study, we report that ultraviolet radiation (UVR), specifically UVB spectra, causes wild-type strains of mice to become highly susceptible to MmuPV1-induced disease...
May 2016: PLoS Pathogens
Anselm Sommer, Felix Kordowski, Joscha Büch, Thorsten Maretzky, Astrid Evers, Jörg Andrä, Stefan Düsterhöft, Matthias Michalek, Inken Lorenzen, Prasath Somasundaram, Andreas Tholey, Frank D Sönnichsen, Karl Kunzelmann, Lena Heinbockel, Christian Nehls, Thomas Gutsmann, Joachim Grötzinger, Sucharit Bhakdi, Karina Reiss
ADAM17, a prominent member of the 'Disintegrin and Metalloproteinase' (ADAM) family, controls vital cellular functions through cleavage of transmembrane substrates. Here we present evidence that surface exposure of phosphatidylserine (PS) is pivotal for ADAM17 to exert sheddase activity. PS exposure is tightly coupled to substrate shedding provoked by diverse ADAM17 activators. PS dependency is demonstrated in the following: (a) in Raji cells undergoing apoptosis; (b) in mutant PSA-3 cells with manipulatable PS content; and (c) in Scott syndrome lymphocytes genetically defunct in their capacity to externalize PS in response to intracellular Ca(2+) elevation...
2016: Nature Communications
Jonathan A Nye, Dana Tudorascu, Fabio Esteves, John R Votaw
Myocardial perfusion studies suffer from artifacts caused by misalignment of the transmission and emission data due to the influences of voluntary and involuntary patient motion. Regardless of 68Ge or respiratory-averaged CT based attenuation correction and good patient cooperation, approximately 21% of perfusion studies exhibit artifacts arising from misalignment that cannot be corrected by manipulating the attenuation acquisition protocol. This misalignment, termed cardiac drift, is caused by slow-moving abdominal cavity contents that reposition the heart in the thorax and appear as myocardial uptake overlying the left CT lung in fused PET/CT images...
2016: Journal of Applied Clinical Medical Physics
Cameron A McConkey, Elizabeth Delorme-Axford, Cheryl A Nickerson, Kwang Sik Kim, Yoel Sadovsky, Jon P Boyle, Carolyn B Coyne
In eutherians, the placenta acts as a barrier and conduit at the maternal-fetal interface. Syncytiotrophoblasts, the multinucleated cells that cover the placental villous tree surfaces of the human placenta, are directly bathed in maternal blood and are formed by the fusion of progenitor cytotrophoblasts that underlie them. Despite their crucial role in fetal protection, many of the events that govern trophoblast fusion and protection from microbial infection are unknown. We describe a three-dimensional (3D)-based culture model using human JEG-3 trophoblast cells that develop syncytiotrophoblast phenotypes when cocultured with human microvascular endothelial cells...
March 2016: Science Advances
Maisem Laabei, Ruth Massey
From the first identification of bacteria as a causative agent of disease, researchers have been developing methods and techniques to understand their pathogenic processes. For decades, this work has been limited to looking at a small number of genetically manipulatable isolates in in vitro assays or animal models of infection. Despite these limitations such work has facilitated the development of successful therapeutic strategies, most notably vaccines that target specific virulence-related features. There are however many antimicrobial resistant pathogens for which vaccination strategies have not worked, as we simply do not know enough about how they cause disease...
August 2016: Current Genetics
Marthe T C Walvoort, Vinita Lukose, Barbara Imperiali
Phosphoglycosyltransferases (PGTs) represent "gatekeeper" enzymes in complex glycan assembly pathways by catalyzing transfer of a phosphosugar from an activated nucleotide diphosphosugar to a membrane-resident polyprenol phosphate. The unique structures of selected nucleoside antibiotics, such as tunicamycin and mureidomycin A, which are known to inhibit comparable biochemical transformations, are exploited as the foundation for the development of modular synthetic inhibitors of PGTs. Herein we present the design, synthesis, and biochemical evaluation of two readily manipulatable modular scaffolds as inhibitors of monotopic bacterial PGTs...
March 7, 2016: Chemistry: a European Journal
Emily R Clark, Donald M Kurtz
We report photosensitized H2 generation using a protein scaffold that nucleates formation of platinum nanoparticles (Pt NPs) and contains "built-in" photosensitizers. The photosensitizers, zinc-protoporphyrin IX or zinc-mesoporphyrin IX (ZnP) were incorporated in place of the naturally occurring heme in the 24-subunit iron storage protein bacterioferritin (Bfr) when the ZnPs were added to the E. coli expression medium. We engineered a stable dimeric Bfr variant with two protein subunits sandwiching a ZnP. Ten glycines were also substituted in place of residues surrounding the vinyl side of the porphyrin in order increase access of solvent and/or redox agents...
January 14, 2016: Dalton Transactions: An International Journal of Inorganic Chemistry
Kartik A Shah, John J Clark, Brittany A Goods, Timothy J Politano, Nicholas J Mozdzierz, Ross M Zimnisky, Rachel L Leeson, J Christopher Love, Kerry R Love
Monoclonal antibodies (mAbs) that bind and neutralize human pathogens have great therapeutic potential. Advances in automated screening and liquid handling have resulted in the ability to discover antigen-specific antibodies either directly from human blood or from various combinatorial libraries (phage, bacteria, or yeast). There remain, however, bottlenecks in the cloning, expression and evaluation of such lead antibodies identified in primary screens that hinder high-throughput screening. As such, "hit-to-lead identification" remains both expensive and time-consuming...
December 2015: Biotechnology and Bioengineering
Qingtao Li, Liangpeng Ge, Wenbing Wan, Junzi Jiang, Wen Zhong, Jun Ouyang, Malcolm Xing
A persistent challenge in tissue engineering is the fabrication of manipulatable scaffolds for implantation in clinical treatments and use in disease models for drug screening. Electrospinning of nanofibrous membranes is an emerging technology in artificial extracellular matrix (ECM) design that can offer precisely tunable microenvironments upon assembly into three-dimensional (3D) scaffolds that mimic the in vivo ECM structure. In this study, we report a facile and versatile strategy for preparing 3D multilayered constructs from Fe3O4/polycaprolactone (PCL)/gelatin nanofibrous membranes...
October 2015: Tissue Engineering. Part C, Methods
Zuhaib Fayaz Bhat, Sunil Kumar, Hina Fayaz Bhat
In vitro meat production is a novel idea of producing meat without involving animals with the help of tissue engineering techniques. This biofabrication of complex living products by using various bioengineering techniques is a potential solution to reduce the ill effects of current meat production systems and can dramatically transform traditional animal-based agriculture by inventing 'animal-free' meat and meat products. Nutrition-related diseases, food borne illnesses, resource use and pollution, and use of farm animals are some serious consequences associated with conventional meat production methods...
May 5, 2015: Critical Reviews in Food Science and Nutrition
Ang Li, Yung-Chih Lai, Seth Figueroa, Tian Yang, Randall B Widelitz, Krzysztof Kobielak, Qing Nie, Cheng Ming Chuong
BACKGROUND: How tissue patterns form in development and regeneration is a fundamental issue remaining to be fully understood. The integument often forms repetitive units in space (periodic patterning) and time (cyclic renewal), such as feathers and hairs. Integument patterns are visible and experimentally manipulatable, helping us reveal pattern formative processes. Variability is seen in regional phenotypic specificities and temporal cycling at different physiological stages. RESULTS: Here we show some cellular/molecular bases revealed by analyzing integument patterns...
August 2015: Developmental Dynamics: An Official Publication of the American Association of Anatomists
Purnima K Wagh, Margaret A Gardner, Xiaolan Ma, Melissa Callahan, John M Shannon, Susan E Wert, Yoav H Messinger, Louis P Dehner, D Ashley Hill, Kathryn A Wikenheiser-Brokamp
Inherited syndromes provide unique opportunities to identify key regulatory mechanisms governing human disease. We previously identified germline loss-of-function DICER1 mutations in a human syndrome defined by the childhood lung neoplasm pleuropulmonary blastoma (PPB), which arises during lung development. DICER1 regulates many biological processes critical in development and disease pathogenesis. Significant challenges in defining the role of DICER1 in human disease are identifying cause-effect relationships and generating manipulatable systems that model the complexity of organ development and disease pathogenesis...
May 2015: Journal of Pathology
Sally A Linkenauger, Michael N Geuss, Jeanine K Stefanucci, Markus Leyrer, Beth H Richardson, Dennis R Proffitt, Heinrich H Bülthoff, Betty J Mohler
The hand is a reliable and ecologically useful perceptual ruler that can be used to scale the sizes of close, manipulatable objects in the world in a manner similar to the way in which eye height is used to scale the heights of objects on the ground plane. Certain objects are perceived proportionally to the size of the hand, and as a result, changes in the relationship between the sizes of objects in the world and the size of the hand are attributed to changes in object size rather than hand size. To illustrate this notion, we provide evidence from several experiments showing that people perceive their dominant hand as less magnified than other body parts or objects when these items are subjected to the same degree of magnification...
November 2014: Psychological Science
Biswajit Mukherjee
Nanosize materials provide hopes, speculations and chances for an unprecedented change in drug delivery in near future. Nanotechnology is an emerging field to produce nanomaterials for drug delivery that can offer a new tool, opportunities and scope to provide more focused and fine-tuned treatment of diseases at a molecular level, enhancing the therapeutic potential of drugs so that they become less toxic and more effective. Nanodimensional drug delivery systems are of great scientific interest as they project their tremendous utility because of their capability of altering biodistribution of therapeutic agents so that they can concentrate more in the target tissues...
2013: Current Pharmaceutical Biotechnology
Joe Alcock, Carlo C Maley, C Athena Aktipis
Microbes in the gastrointestinal tract are under selective pressure to manipulate host eating behavior to increase their fitness, sometimes at the expense of host fitness. Microbes may do this through two potential strategies: (i) generating cravings for foods that they specialize on or foods that suppress their competitors, or (ii) inducing dysphoria until we eat foods that enhance their fitness. We review several potential mechanisms for microbial control over eating behavior including microbial influence on reward and satiety pathways, production of toxins that alter mood, changes to receptors including taste receptors, and hijacking of the vagus nerve, the neural axis between the gut and the brain...
October 2014: BioEssays: News and Reviews in Molecular, Cellular and Developmental Biology
Marc D Bullock, Max Mellone, Karen M Pickard, Abdulkadir Emre Sayan, Richard Mitter, John N Primrose, Graham K Packham, Gareth Thomas, Alexander H Mirnezami
Invading colorectal cancer (CRC) cells have acquired the capacity to break free from their sister cells, infiltrate the stroma, and remodel the extracellular matrix (ECM). Characterizing the biology of this phenotypically distinct group of cells could substantially improve our understanding of early events during the metastatic cascade. Tumor invasion is a dynamic process facilitated by bidirectional interactions between malignant epithelium and the cancer associated stroma. In order to examine cell-specific responses at the tumor stroma-interface we have combined organotypic co-culture and laser micro-dissection techniques...
2014: Journal of Visualized Experiments: JoVE
Ryan A Koppes, Douglas M Swank, David T Corr
Force depression (FD) is a decrease in isometric force following active muscle shortening. Despite being well characterized experimentally, its underlying mechanism remains unknown. To develop a new, genetically manipulatable experimental model that would greatly improve our ability to study the underlying mechanism(s) of FD, we tested the Drosophila jump muscle for classical FD behavior. Steady-state force generation following active shortening decreased by 2, 8, and 11% of maximum isometric force with increasing shortening amplitudes of 5, 10, and 20% of optimal fiber length, and decreased by 11, 8, and 5% with increasing shortening velocities of 4, 20, and 200% of optimal fiber length per second...
June 15, 2014: Journal of Applied Physiology
Vladimir Zlateski, Roland Fuhrer, Fabian M Koehler, Scott Wharry, Martin Zeltner, Wendelin J Stark, Thomas S Moody, Robert N Grass
In the pursuit of robust and reusable biocatalysts for industrial synthetic chemistry, nanobiotechnology is currently taking a significant part. Recently, enzymes have been immobilized on different nanoscaffold supports. Carbon coated metallic nanoparticles were found to be a practically useful support for enzyme immobilization due to their large surface area, high magnetic saturation, and manipulatable surface chemistry. In this study carbon coated cobalt nanoparticles were chemically functionalized (diazonium chemistry), activated for bioconjugation (N,N-disuccinimidyl carbonate), and subsequently used in enzyme immobilization...
April 16, 2014: Bioconjugate Chemistry
Meng Cheng, Duoming Wang, Zhaoru Sun, Jing Zhao, Rong Yang, Guole Wang, Wei Yang, Guibai Xie, Jing Zhang, Peng Chen, Congli He, Donghua Liu, Limei Xu, Dongxia Shi, Enge Wang, Guangyu Zhang
Manipulation of an isolated water nanodroplet (WN) on certain surfaces is important to various nanofluidic applications but challenging. Here we present a digital nanofluidic system based on a graphene/water/mica sandwich structure. In this architecture, graphene provides a flexible protection layer to isolate WNs from the outside environment, and a monolayer ice-like layer formed on the mica surface acts as a lubricant layer to allow these trapped WNs to move on it freely. In combination with scanning probe microscope techniques, we are able to move, merge, and separate individual water nanodroplets in a controlled manner...
April 22, 2014: ACS Nano
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