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Proof of concept

Susumu Ishiguro, Nabil A Alhakamy, Deepthi Uppalapati, Jennifer Delzeit, Cory J Berkland, Masaaki Tamura
To evaluate the potential of cell-penetrating peptide-based delivery of apoptosis-inducer gene in cancer therapy, a modified HIV-1 TAT peptide (dimerized TAT peptide, dTAT) was studied. The dTAT and plasmid DNA (pDNA) complexes (dTAT-pDNA) were condensed using calcium chloride (dTAT-pDNA-Ca(2+)). This simple nonviral formulation approach showed high levels of gene expression in vitro without any cytotoxicity. In mouse studies, a single intratracheal (IT) aerosol spray or 2 intravenous (IV) injections of the dTAT, apoptosis-inducer gene, angiotensin II type 2 receptor (AT2R), and Ca(2+) complexes (dTAT-pAT2R-Ca(2+)) significantly attenuated the acutely growing mouse Lewis lung carcinoma allografts in mouse lungs...
October 18, 2016: Journal of Pharmaceutical Sciences
Seshasailam Venkateswaran, Maria Angélica Luque-González, Mavys Tabraue-Chávez, Mario Antonio Fara, Barbara López-Longarela, Victoria Cano-Cortes, Francisco Javier López-Delgado, Rosario María Sánchez-Martín, Hugh Ilyine, Mark Bradley, Salvatore Pernagallo, Juan José Díaz-Mochón
Over the last decade, circulating microRNAs have received attention as diagnostic and prognostic biomarkers. In particular, microRNA122 has been demonstrated to be an early and more sensitive indicator of drug-induced liver injury than the widely used biomarkers such as alanine aminotransferase and aspartate aminotransferase. Recently, microRNA122 has been used in vitro to assess the cellular toxicity of new drugs and as a biomarker for the development of a rapid test for drug overdose/liver damage. In this proof-of-concept study, we report a PCR-free and label-free detection method that has a limit of detection (3 standard deviations) of 15 fmoles of microRNA122, by integrating a dynamic chemical approach for "Single Nucleobase Labelling" with a bead-based platform (Luminex(®)) thereby, in principle, demonstrating the exciting prospect of rapid and accurate profiling of any microRNAs related to diseases and toxicology...
December 1, 2016: Talanta
Seokjin Suh, Xiangpeng Meng, Sanghoon Ko
The objective of this study was to develop different-sized chitosan nanoparticles as CO2-based food quality indicators. Chitosan nanoparticles were fabricated with different sizes (small, 300nm; medium, 500nm; and large, 1000nm) by ionic gelation. To investigate the performance of chitosan nanoparticles as CO2 indicators, they were suspended in aqueous solution at pH 8.0. Changes in the pH and absorbance of the suspension were measured over time, the absorbance at the transition appearance time being calculated using the inverse-Hill function...
December 1, 2016: Talanta
Pavel Kubáň, Knut Fredrik Seip, Astrid Gjelstad, Stig Pedersen-Bjergaard
This work investigated selective micro-electromembrane extractions (μ-EMEs) of the colored indicators metanil yellow and congo red (visual proof-of-principle) and the small drug substances nortriptyline, papaverine, mianserin, and citalopram (model analytes) based on their acid-base strength. With two free liquid membranes (FLMs), the target analytes were extracted from aqueous donor solution, across FLM 1 (1-pentanol, 1-ethyl-2-nitrobenzene (ENB) or 4-nitrocumene (4-NC)), into aqueous acceptor solution 1, further across FLM 2 (1-pentanol, ENB or 4-NC), and finally into aqueous acceptor solution 2...
November 2, 2016: Analytica Chimica Acta
Brenda F Narice, Nicola H Green, Sheila MacNeil, Dilly Anumba
BACKGROUND: During labour, the cervix undergoes a series of changes to allow the passage of the fetoplacental unit. While this visible transformation is well-described, the underlying and causative microscopic changes, in which collagen plays a major role, are poorly understood and difficult to visualise. Recent studies in mice and humans have shown that Second Harmonic Generation (SHG) microscopy, a non-destructive imaging technique, can detect changes in the cervical collagen. However, the question of whether SHG can identify changes in the arrangement of cervical collagen at different physiological stages still needs addressing...
October 21, 2016: Reproductive Biology and Endocrinology: RB&E
Alexis K Huynh, Martin L Lee, Melissa M Farmer, Lisa V Rubenstein
BACKGROUND: Stepped wedge designs have gained recognition as a method for rigorously assessing implementation of evidence-based quality improvement interventions (QIIs) across multiple healthcare sites. In theory, this design uses random assignment of sites to successive QII implementation start dates based on a timeline determined by evaluators. However, in practice, QII timing is often controlled more by site readiness. We propose an alternate version of the stepped wedge design that does not assume the randomized timing of implementation while retaining the method's analytic advantages and applying to a broader set of evaluations...
October 21, 2016: BMC Medical Research Methodology
Giulia Suarato, Seong-Il Lee, Weiyi Li, Sneha Rao, Tanvir Khan, Yizhi Meng, Maya Shelly
During mammalian embryonic development, neurons polarize to create distinct cellular compartments of axon and dendrite that inherently differ in form and function, providing the foundation for directional signaling in the nervous system. Polarization results from spatio-temporal segregation of specific proteins' activities to discrete regions of the neuron to dictate axonal vs. dendritic fate. We aim to manipulate axon formation by directed subcellular localization of crucial intracellular protein function...
October 8, 2016: Biomaterials
Christopher D Morrone, Lynsie A M Thomason, Mary E Brown, Isabelle Aubert, JoAnne McLaurin
Although it is recognized that multi-drug therapies may be necessary to combat AD, there is a paucity of preclinical proof of concept studies. We present a combination treatment paradigm, which temporally affects different aspects of Alzheimer's disease (AD)-like pathology, specifically Aβ-toxicity and neurogenesis. At early stages of AD-like pathology, in TgCRND8 mice, we found that combating Aβ pathology with scyllo-inositol ameliorated deficits in neurogenesis. Older TgCRND8 mice with established amyloid load had decreased progenitor cell proliferation and survival compared to non-transgenic mice, regardless of scyllo-inositol treatment...
2016: PloS One
Pavel Damborský, Katarzyna M Koczula, Andrea Gallotta, Jaroslav Katrlík
Lateral flow assays (LFAs) enable the simple and rapid detection and quantification of analytes and is popular for point-of-care (PoC), point-of-use and outdoor testing applications. LFAs typically depend on antibody or nucleic acid based recognition. We present the innovative concept of a LFA using lectins in the role of the biorecognition element. Lectins are a special kind of glycan-binding protein and the lectin-based LFA herein described was developed for the determination of the glycosylation of free prostate specific antigen (PSA)...
October 21, 2016: Analyst
Ludovic Enkler, Delphine Richer, Anthony L Marchand, Dominique Ferrandon, Fabrice Jossinet
Among Candida species, the opportunistic fungal pathogen Candida glabrata has become the second most common causative agent of candidiasis in the world and a major public health concern. Yet, few molecular tools and resources are available to explore the biology of C. glabrata and to better understand its virulence during infection. In this study, we describe a robust experimental strategy to generate loss-of-function mutants in C. glabrata. The procedure is based on the development of three main tools: (i) a recombinant strain of C...
October 21, 2016: Scientific Reports
Sasha C Voss, Paola Magni, Ian Dadour, Christian Nansen
Forensic entomology is primarily concerned with the estimation of time since death and involves determination of the age of immature insects colonising decomposing remains. Accurate age determination of puparia is usually accomplished by dissection, which means destructive sampling of evidence. As part of improving abilities to correctly identify species and developmental age, it is highly desirable to have available non-destructive methods. In this study, we acquired external hyperspectral imaging (HSI) data (77 spectral bands, 389-892 nm) from the dorsal and ventral sides of individual puparia of two species of blowfly (Diptera: Calliphoridae), Calliphora dubia Macquart 1855 and Chrysomya rufifacies Macquart 1842...
October 21, 2016: International Journal of Legal Medicine
Jean-Marie E P Cols, Cameron E Taylor, Kevin J Gagnon, Simon J Teat, Ruaraidh D McIntosh
The synthesis and full characterisation of four discrete tetrametallic titanium complexes is reported. These well-defined compounds are isostructural in the solid state and share the same general formula: Ti4(μ-O)2L4 (L = 1, 2, 3 or 4). Using a combination of NMR techniques the complexes are found to be stable in solution, even at elevated temperatures. Further studies show that the carboxylate moieties of the supporting amine bis(phenolate) ligands can be displaced by a more strongly coordinating solvent...
October 21, 2016: Dalton Transactions: An International Journal of Inorganic Chemistry
Laleh Majari Kasmaee, Asghar Aryanfar, Zarui Chikneyan, Michael R Hoffmann, Agustín J Colussi
Understanding the mechanism of formation of solid-electrolyte interphases (SEI) is key to the prospects of lithium metal batteries (LMB). Here, we investigate via cyclic voltammetry, impedance spectroscopy and chronoamperometry the role of kinetics in controlling the properties of the SEI generated from the reduction of propylene carbonate (PC, a typical solvent in LMB). Our observations are consistent with the operation of a radical chain PC electropolymerization into polymer units whose complexity increases at lower initiation rates...
September 16, 2016: Chemical Physics Letters
Yu Sun, Vaibhav A Narayan, Gayle M Wittenberg
BACKGROUND: Side effects, or the adverse effects of drugs, contain important clinical phenotypic information that may be useful in predicting novel or unknown targets of a drug. It has been suggested that drugs with similar side-effect profiles may share common targets. The diagnostic class, Major Depressive Disorder, is increasingly viewed as being comprised of multiple depression subtypes with different biological root causes. One 'type' of depression generating substantial interest today focuses on patients with high levels of inflammatory burden, indicated by elevated levels of C-reactive proteins (CRP) and pro-inflammatory cytokines such as interleukin 6 (IL-6)...
October 21, 2016: BMC Pharmacology & Toxicology
Kun Yang, Yun Gao, Mingfu Yang, Zuoshang Xu, Qian Chen
Because the function of most non-coding (nc) RNAs is unknown, Cre-lox transgenic mice are useful tools to determine their functions in a tissue or developmental stage-specific manner. However, the technology faces challenges because expression of ncRNA-transgene lacks protein product. No antibody or peptide-tag can be used to trace ncRNA expression in mouse tissues in real time. Furthermore, transgene integration at different locus or orientations in the genome may result in recombination of genomic fragments in the Cre-lox system...
October 20, 2016: Connective Tissue Research
Bratindranath Mukherjee, Swagotom Sarker, Eric Crone, Pawan Pathak, Vaidyanathan Ravi Subramanian
This work presents a hitherto unreported approach to assemble a 1D oxide-1D chalcogenide heterostructured photoactive film. As a representative system, bismuth (Bi), a low melting point catalyst, facilitated 1D CdSe nanowires are directly grown on anodized 1D-TiO2 nanotube (T_NT). A combination of the reductive successive-ionic-layer-adsorption-reaction (R-SILAR) and the solution-liquid-solid (S-L-S) approach is implemented towards the assembly of this heterostructure, reported in this 1D-1D form for the first time...
October 20, 2016: ACS Applied Materials & Interfaces
Chao-Chen Lin, Hsin-Fang Hsu, Peter Jomo Walla
The fusion of two opposing membranes is essential in biological functions such as fertilization, viral entry, membrane trafficking and synaptic transmission. Before the membrane bilayers are fully connected, at some stage a hemifusion intermediate-when the outer leaflets are merged but not the inner leaflets-is formed. However, the position of hemifusion in the energy landscape and the duration of it vary and have not been fully mapped out. To date, there has not been a way to differentiate lipid mixing of the two leaflets directly in a single experiment...
October 20, 2016: Journal of Physical Chemistry. B
Sonia Fieulaine, Rodolphe Alves de Sousa, Laure Maigre, Karim Hamiche, Mickael Alimi, Jean-Michel Bolla, Abbass Taleb, Alexis Denis, Jean-Marie Pagès, Isabelle Artaud, Thierry Meinnel, Carmela Giglione
Peptide deformylase (PDF) is considered an excellent target to develop antibiotics. We have performed an extensive characterization of a new PDF from the pathogen Streptococcus agalactiae, showing properties similar to other known PDFs. S. agalactiae PDF could be used as PDF prototype as it allowed to get complete sets of 3-dimensional, biophysical and kinetic data with virtually any inhibitor compound. Structure-activity relationship analysis with this single reference system allowed us to reveal distinct binding modes for different PDF inhibitors and the key role of a hydrogen bond in potentiating the interaction between ligand and target...
October 20, 2016: Scientific Reports
Jennifer E Adair, Timothy Waters, Kevin G Haworth, Sara P Kubek, Grant D Trobridge, Jonah D Hocum, Shelly Heimfeld, Hans-Peter Kiem
Haematopoietic stem cell (HSC) gene therapy has demonstrated potential to treat many diseases. However, current state of the art requires sophisticated ex vivo gene transfer in a dedicated Good Manufacturing Practices facility, limiting availability. An automated process would improve the availability and standardized manufacture of HSC gene therapy. Here, we develop a novel program for semi-automated cell isolation and culture equipment to permit complete benchtop generation of gene-modified CD34(+) blood cell products for transplantation...
October 20, 2016: Nature Communications
Yinghe Huo, Renske D Veldhuizen, Desiree M van der Heijde, Nick J Besselink, Johannes W G Jacobs, Jacob M van Laar, Max A Viergever, Koen L Vincken, Floris P Lafeber, Maria J H de Hair
OBJECTIVES: To compare as proof of concept the sensitivity to change of automated quantification of radiographic wrist and hand joint space width (JSW) with scoring JSW according to the Sharp/van der Heijde scoring method (SHS) in two strategy groups of a treat-to-target and tight-control early rheumatoid arthritis (RA) study. METHODS: Digital radiographs were assessed for JSW changes of 134 patients of the 236 patients participating in the second Computer Assisted Management in Early Rheumatoid Arthritis trial, of whom both baseline and year 2 radiographs were available (year 1 radiographs n=125)...
September 2016: Clinical and Experimental Rheumatology
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