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https://www.readbyqxmd.com/read/29054027/sapofectosid-ensuring-non-toxic-and-effective-dna-and-rna-delivery
#1
Simko Sama, Gerold Jerz, Peter Schmieder, Eric Woith, Matthias F Melzig, Alexander Weng
Different methods are being deployed for non-viral DNA/RNA delivery. However non-viral formulations for DNA/RNA-delivery are often accompanied by severe toxicity and thus low efficiency. Particular costly cell culture media are required as well. Here we introduce sapofection as a valuable enhancing method for non-viral DNA/RNA delivery. Sapofection is based on the application of DNA/RNA nanoplexes and sapofectosid, a plant derived natural transfection reagent. Sapofectosid was produced from plant raw material by chromatographic methods and characterized by tandem mass spectrometry and intensive one and two dimensional NMR-spectroscopy...
October 17, 2017: International Journal of Pharmaceutics
https://www.readbyqxmd.com/read/29053825/pdm3-a-pentatricopeptide-repeat-containing-protein-affects-chloroplast-development
#2
Jian Zhang, Jianwei Xiao, Yuqian Li, Bodan Su, Huimin Xu, Xiaoyi Shan, Chengwei Song, Jianbo Xie, Ruili Li, Robert Sharwood
The chloroplast, as the photosynthetic organelle of plants, plays a crucial role in plant development. Extensive studies have been conducted on chloroplast development; however, the related regulatory mechanism still remains elusive. Here, we characterized a mutant with defective chloroplasts in Arabidopsis, termed pigment-defective mutant3 (pdm3), which exhibits a distinct albino phenotype in leaves, eventually leading to pdm3 seedling lethality under autotrophic growth conditions. Electron microscopy demonstrated that the number of thylakoids was reduced and the structure of those thylakoids was disrupted in the pdm3 mutant, which eventually led to the breakdown of chloroplasts...
October 19, 2017: Journal of Experimental Botany
https://www.readbyqxmd.com/read/29053699/evaluation-of-synapse-density-in-hippocampal-rodent-brain-slices
#3
Faye McLeod, Aude Marzo, Marina Podpolny, Soledad Galli, Patricia Salinas
In the brain, synapses are specialized junctions between neurons, determining the strength and spread of neuronal signaling. The number of synapses is tightly regulated during development and neuronal maturation. Importantly, deficits in synapse number can lead to cognitive dysfunction. Therefore, the evaluation of synapse number is an integral part of neurobiology. However, as synapses are small and highly compact in the intact brain, the assessment of absolute number is challenging. This protocol describes a method to easily identify and evaluate synapses in hippocampal rodent slices using immunofluorescence microscopy...
October 6, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29053691/quantitative-whole-mount-immunofluorescence-analysis-of-cardiac-progenitor-populations-in-mouse-embryos
#4
Evan Bardot, Nikos Tzavaras, Deanna L Benson, Nicole C Dubois
The use of ever-advancing imaging techniques has contributed broadly to our increased understanding of embryonic development. Pre-implantation development and organogenesis are two areas of research that have benefitted greatly from these advances, due to the high quality of data that can be obtained directly from imaging pre-implantation embryos or ex vivo organs. While pre-implantation embryos have yielded data with especially high spatial resolution, later stages have been less amenable to three-dimensional reconstruction...
October 12, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29053678/assay-for-phosphorylation-and-microtubule-binding-along-with-localization-of-tau-protein-in-colorectal-cancer-cells
#5
Md Nazmul Huda, Erdenedolgor Erdene-Ochir, Cheol-Ho Pan
The microtubule-associated protein tau is a neuronal protein that localizes mostly in axons. Generally tau is essential for normal neuronal functioning because it is involved in microtubule assembly and stabilization. Besides neurons, tau is expressed in human breast, prostate, gastric, colorectal, and pancreatic cancers where it shows nearly similar structure and exerts similar functions as the neuronal tau. The amount of tau and its phosphorylation can change its function as a stabilizer of microtubules, and lead to the development of paired helical filaments in different neurodegenerative disorders, such as Alzheimer's disease...
October 10, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29053558/in-vivo-confocal-microscopy-reveals-corneal-reinnervation-after-treatment-of-neurotrophic-keratopathy-with-corneal-neurotization
#6
Simon S M Fung, Joseph Catapano, Uri Elbaz, Ronald M Zuker, Gregory H Borschel, Asim Ali
PURPOSE: To document the presence and location of new sensory nerve fibers after corneal neurotization using in vivo confocal microscopy (IVCM) in 2 patients with neurotrophic keratopathy (NK). METHODS: Two patients with unilateral advanced NK received corneal neurotization to surgically reinnervate the cornea. IVCM was used to identify subbasal nerve fibers and document corneal reinnervation. In 1 patient (case 1), IVCM was performed before and after corneal neurotization; in the second patient (case 2), IVCM was performed after neurotization and corneal transplantation...
October 17, 2017: Cornea
https://www.readbyqxmd.com/read/29053406/super-resolution-nanoscopy-imaging-applied-to-dna-double-strand-breaks
#7
Sofia D'Abrantes, Sarah Gratton, Pamela Reynolds, Verena Kriechbaumer, Joseph McKenna, Stephen Barnard, Dave Clarke, Stanley W Botchway
Genomic deoxyribonucleic acid (DNA) is continuously being damaged by endogenous processes such as metabolism or by exogenous events such as radiation. The specific phosphorylation of histone H2AX on serine residue 139, described as γ-H2AX, is an excellent indicator or marker of DNA double-strand breaks (DSBs). The yield of γ-H2AX (foci) is shown to have some correlation with the dose of radiation or other DSB-causing agents. However, there is some discrepancy in the DNA DSB foci yield among imaging and other methods such as gel electrophoresis...
October 20, 2017: Radiation Research
https://www.readbyqxmd.com/read/29052767/acute-regulation-of-multidrug-resistance-associated-protein-2-localization-and-activity-by-camp-and-estradiol-17%C3%AE-d-glucuronide-in-rat-intestine-and-caco-2-cells
#8
Guillermo Nicolás Tocchetti, Agostina Arias, Maite Rocío Arana, Juan Pablo Rigalli, Camila Juliana Domínguez, Felipe Zecchinati, María Laura Ruiz, Silvina Stella Maris Villanueva, Aldo Domingo Mottino
Multidrug resistance-associated protein 2 (MRP2) is an ATP-dependent transporter expressed at the brush border membrane of the enterocyte that confers protection against absorption of toxicants from foods or bile. Acute, short-term regulation of intestinal MRP2 activity involving changes in its apical membrane localization was poorly explored. We evaluated the effects of dibutyryl-cAMP (db-cAMP), a permeable analog of cAMP, and estradiol-17β-D-glucuronide (E217G), an endogenous derivative of estradiol, on MRP2 localization and activity using isolated rat intestinal sacs and Caco-2 cells, a model of human intestinal epithelium...
October 20, 2017: Archives of Toxicology
https://www.readbyqxmd.com/read/29052755/validation-of-an-easily-applicable-three-dimensional-immunohistochemical-imaging-method-for-a-mouse-brain-using-conventional-confocal-microscopy
#9
Tetsuhiro Kakimoto
Histological analysis has been largely confined to two-dimensional analysis of thin tissue sections, hampering detailed understanding of three-dimensional cellular distribution in biological tissues. Tissue optical clearing methods enable three-dimensional histological analysis by rendering tissues transparent and suitable for microscopic detection of the fluorescent signals inside. Despite their great potential in histological research, the tissue clearing methods are not readily accessible to many researchers because of hazardous chemicals, complicated protocols and advanced microscopy...
October 20, 2017: Histochemistry and Cell Biology
https://www.readbyqxmd.com/read/29051962/structural-disorder-filament-growth-and-self-poisoning-in-short-rods-confined-onto-a-flat-wall
#10
Yongxiang Gao, Violet Farkas, Roel P A Dullens, Dirk G A L Aarts
Confocal microscopy was used to directly observe the structural coarsening of the first layer of short colloidal rods sedimented onto a flat wall. Based on an image analysis algorithm we devised, quantitative information on the location, orientation and length of each particle can be extracted with high precision. At high density the system undergoes structural arrest, and becomes trapped in a disordered state of randomly arranged filaments that are composed of side-by-side aligned rods. The frustration of structural order is signalled by a new peak that emerges in the radial distribution function...
October 20, 2017: Soft Matter
https://www.readbyqxmd.com/read/29051762/specific-pathogen-recognition-by-multiple-innate-immune-sensors-in-an-invertebrate
#11
Guillaume Tetreau, Silvain Pinaud, Anaïs Portet, Richard Galinier, Benjamin Gourbal, David Duval
Detection of pathogens by all living organisms is the primary step needed to implement a coherent and efficient immune response. This implies a mediation by different soluble and/or membrane-anchored proteins related to innate immune receptors called PRRs (pattern-recognition receptors) to trigger immune signaling pathways. In most invertebrates, their roles have been inferred by analogy to those already characterized in vertebrate homologs. Despite the induction of their gene expression upon challenge and the presence of structural domains associated with the detection of pathogen-associated molecular patterns in their sequence, their exact role in the induction of immune response and their binding capacity still remain to be demonstrated...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/29051198/abscisic-acid-induced-reactive-oxygen-species-are-modulated-by-flavonols-to-control-stomata-aperture
#12
Justin Watkins, Jordan M Chapman, Gloria K Muday
Abscisic acid (ABA) increases reactive oxygen species (ROS) in guard cells to close Arabidopsis stomata. In Solanum lycopersicum we find that ABA increased ROS is followed by stomatal closure and both responses are blocked by inhibitors of ROS-producing respiratory burst oxidase enzymes. ABA-induced ROS sensor fluorescence accumulates in the nucleus, chloroplasts, and in endomembranes. Accumulation of flavonol antioxidants in guard cells, but not surrounding pavement cells, were visualized by confocal microscopy using a flavonol-specific fluorescent dye...
October 19, 2017: Plant Physiology
https://www.readbyqxmd.com/read/29051187/in-vivo-magnetofection-a-novel-approach-for-targeted-topical-delivery-of-nucleic-acids-for-rectoanal-motility-disorders
#13
Jagmohan Singh, Ipsita Mohanty, Satish Rattan
In these studies, we developed a novel approach of in vivo magnetofection for localized delivery of nucleic acids such as micro-RNA-139-p (miR-139-5p, known to target Rho kinase2) to the circular smooth muscle layer of the internal anal sphincter (IAS). The IAS tone is known to play a major role in the rectoanal continence via activation of RhoA-associated kinase (RhoA/ROCK2). These studies established an optimized protocol for efficient gene delivery using an assembly of equal volumes of in vivo PolyMag and miR139-5p or anti-miR-139-5p (100 nM each) injected in the circular smooth muscle layer in the pin-pointed areas of the rat perianal region, and then incubated for 20 minutes under magnetic field...
October 19, 2017: American Journal of Physiology. Gastrointestinal and Liver Physiology
https://www.readbyqxmd.com/read/29051061/precise-laser-poration-to-control-drug-delivery-into-and-through-human-nail
#14
Simon Vanstone, Sarah F Cordery, James M Stone, Sergey N Gordeev, Richard H Guy
Drug treatment of diseases of the human nail remains a difficult challenge; topical therapy, in particular, is limited by very poor transport of active agents across the nail itself. The objective of this research was to examine the potential of controlled, and fibre-optic delivered, femtosecond laser light pulses to provide new pathways and opportunities for drug access to targets within and beneath the nail plate. Optical, confocal fluorescence and scanning electron microscopies demonstrated partial and complete laser poration of human nail samples, with the energy per pore and the exposure duration being the key modulating parameters that determined the extent of ablation achieved...
October 16, 2017: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/29050619/novel-starch-based-emulsion-gels-and-emulsion-microgel-particles-design-structure-and-rheology
#15
Ophelie Torres, Nidia Mercado Tena, Brent Murray, Anwesha Sarkar
Novel starch-based emulsion microgel particles were designed using a facile top-down shear-induced approach. The emulsion droplets were stabilized using octenyl succinic anhydride (OSA) modified starch and incorporated into heat-treated and sheared native starch gels, forming emulsion gels. Using gelation kinetics and small deformation rheological measurements of sheared native starch gels and emulsion gels, OSA starch-stabilized emulsion droplets were demonstrated to act as "active fillers". By varying native starch concentrations (15-20wt%) and oil fractions (5-20wt%), optimal concentrations for the formation of emulsion microgel particles were identified...
December 15, 2017: Carbohydrate Polymers
https://www.readbyqxmd.com/read/29050594/novel-bioactive-surface-functionalization-of-bacterial-cellulose-membrane
#16
Wei Shao, Jimin Wu, Hui Liu, Shan Ye, Lei Jiang, Xiufeng Liu
Bacterial cellulose (BC) membrane is a promising biopolymer which can be used for tissue implants, wound healing, and drug delivery due to its unique properties, such as high crystallinity, high mechanical strength, ultrafine fiber network structure, good water holding capacity and biocompatibility. However, BC does not intrinsically present antibacterial properties. In the present study, functionalized BC membranes were prepared. FTIR, SEM and XPS were used to characterize the chemical composition and surface morphology...
December 15, 2017: Carbohydrate Polymers
https://www.readbyqxmd.com/read/29050444/treatment-of-ichthyosis-vulgaris-with-a-urea-based-emulsion-videodermatoscopy-and-confocal-microscopy-evaluation
#17
Clara Benintende, Simona Boscaglia, Franco Dinotta, Francesco Lacarrubba, Giuseppe Micali
BACKGROUND: Ichthyosis vulgaris is a common disorder of keratinization caused by mutations in the filaggrin gene and clinically characterized by variable degree of xerosis. METHODS: Five patients affected by ichthyosis vulgaris and moderate to severe xerosis of the lower limbs, were treated twice daily for 30 days with an emulsion containing 10% urea, ceramides, and natural moisturizing factors (NMF). Evaluation was performed at baseline and at the end of treatment by clinical examination, Visual Analogue Scale to quantify itch, videodermatoscopy (VD), and reflectance confocal microscopy (RCM)...
December 2017: Giornale Italiano di Dermatologia e Venereologia: Organo Ufficiale, Società Italiana di Dermatologia e Sifilografia
https://www.readbyqxmd.com/read/29050189/-clinical-features-and-treatments-of-retrocorneal-fungal-infection
#18
J Cheng, H L Zhai, J Y Wang, L X Xie
Objective: To evaluate the clinical features of retrocorneal fungal infection and the therapeutic effects. Methods: This was a retrospective, noncomparative study of nine patients with retrocorneal fungal infection and an intact corneal epithelium treated at Qingdao Eye Hospital. The history, clinical features, diagnostic methods, pathogens and therapeutic effects were analyzed. Results: Five patients had a history of trauma by plant, two had a non-plant injury, and two had unidentified reasons. The duration between the initial onset and the first visit to our hospital was 7...
October 11, 2017: [Zhonghua Yan Ke za Zhi] Chinese Journal of Ophthalmology
https://www.readbyqxmd.com/read/29049733/tunneling-nanotubes-are-novel-cellular-structures-that-communicate-signals-between-trabecular-meshwork-cells
#19
Kate E Keller, John M Bradley, Ying Ying Sun, Yong-Feng Yang, Ted S Acott
Purpose: The actin cytoskeleton of trabecular meshwork (TM) cells plays a role in regulating aqueous humor outflow. Many studies have investigated stress fibers, but F-actin also assembles into other supramolecular structures including filopodia. Recently, specialized filopodia called tunneling nanotubes (TNTs) have been described, which communicate molecular signals and organelles directly between cells. Here, we investigate TNT formation by TM cells. Methods: Human TM cells were labeled separately with the fluorescent dyes, DiO and DiD, or with mitochondrial dye...
October 1, 2017: Investigative Ophthalmology & Visual Science
https://www.readbyqxmd.com/read/29049729/myocilin-regulates-metalloprotease-2-activity-through-interaction-with-timp3
#20
Myung Kuk Joe, Raquel L Lieberman, Naoki Nakaya, Stanislav I Tomarev
Purpose: To elucidate functions of wild-type myocilin, a secreted glycoprotein associated with glaucoma. Methods: Lysates of mouse eyes were used for immunoprecipitation with affinity-purified antibodies against mouse myocilin. Shotgun proteomic analysis was used for the identification of proteins interacting with myocilin. Colocalization of myocilin and tissue inhibitor of metalloproteinases 3 (TIMP3) in different eye structures was investigated by a multiplex fluorescent in situ hybridization and immunofluorescent labeling with subsequent confocal microscopy...
October 1, 2017: Investigative Ophthalmology & Visual Science
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