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https://www.readbyqxmd.com/read/29155886/insights-into-the-mechanism-of-isoenzyme-specific-signal-peptide-peptidase-mediated-translocation-of-heme-oxygenase
#1
Bianca Schaefer, Kohji Moriishi, Soenke Behrends
It has recently been shown that signal peptide peptidase (SPP) can catalyze the intramembrane cleavage of heme oxygenase-1 (HO-1) that leads to translocation of HO-1 into the cytosol and nucleus. While there is consensus that translocated HO-1 promotes tumor progression and drug resistance, the physiological signals leading to SPP-mediated intramembrane cleavage of HO-1 and the specificity of the process remain unclear. In this study, we used co-immunoprecipitation and confocal laser scanning microscopy to investigate the translocation mechanism of HO-1 and its regulation by SPP...
2017: PloS One
https://www.readbyqxmd.com/read/29155761/advanced-confocal-microscopy-techniques-to-study-protein-protein-interactions-and-kinetics-at-dna-lesions
#2
Soňa Legartová, Jana Suchánková, Jana Krejčí, Alena Kovaříková, Eva Bártová
Local microirradiation with lasers represents a useful tool for studies of DNA-repair-related processes in live cells. Here, we describe a methodological approach to analyzing protein kinetics at DNA lesions over time or protein-protein interactions on locally microirradiated chromatin. We also show how to recognize individual phases of the cell cycle using the Fucci cellular system to study cell-cycle-dependent protein kinetics at DNA lesions. A methodological description of the use of two UV lasers (355 nm and 405 nm) to induce different types of DNA damage is also presented...
November 12, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29155738/imaging-amyloid-tissues-stained-with-luminescent-conjugated-oligothiophenes-by-hyperspectral-confocal-microscopy-and-fluorescence-lifetime-imaging
#3
Sofie Nyström, Marcus Bäck, K Peter R Nilsson, Per Hammarström
Proteins that deposit as amyloid in tissues throughout the body can be the cause or consequence of a large number of diseases. Among these we find neurodegenerative diseases such as Alzheimer's and Parkinson's disease afflicting primarily the central nervous system, and systemic amyloidosis where serum amyloid A, transthyretin and IgG light chains deposit as amyloid in liver, carpal tunnel, spleen, kidney, heart, and other peripheral tissues. Amyloid has been known and studied for more than a century, often using amyloid specific dyes such as Congo red and Thioflavin T (ThT) or Thioflavin (ThS)...
October 20, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29155726/visualization-and-live-imaging-of-oligodendrocyte-organelles-in-organotypic-brain-slices-using-adeno-associated-virus-and-confocal-microscopy
#4
Lauritz Hagen Kennedy, Johanne Egge Rinholm
Neurons rely on the electric insulation and trophic support of myelinating oligodendrocytes. Despite the importance of oligodendrocytes, the advanced tools currently used to study neurons, have only partly been taken on by oligodendrocyte researchers. Cell type-specific staining by viral transduction is a useful approach to study live organelle dynamics. This paper describes a protocol for visualizing oligodendrocyte mitochondria in organotypic brain slices by transduction with adeno-associated virus (AAV) carrying genes for mitochondrial targeted fluorescent proteins under the transcriptional control of the myelin basic protein promoter...
October 23, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29155721/visualizing-macrophage-extracellular-traps-using-confocal-microscopy
#5
Roleen Sharma, Kim M O'Sullivan, Stephen R Holdsworth, Philip G Bardin, Paul T King
A primary method used to define the presence of neutrophil extracellular traps (NETs) is confocal microscopy. We have modified established confocal microscopy methods to visualize macrophage extracellular traps (METs). These extracellular traps are defined by the presence of extracellular chromatin with co-expression of other components such as granule proteases, citrullinated histones, and peptidyl arginase deiminase (PAD). The expression of METs is generally measured after exposure to a stimulus and compared to un-stimulated samples...
October 19, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29155399/green-synthesis-of-anisotropic-gold-nanoparticles-using-hordenine-and-their-antibiofilm-efficacy-against-pseudomonas-aeruginosa
#6
Jobina Rajkumari, Himani Meena, Muralitharan Gangatharan, Siddhardha Busi
Pseudomonas aeruginosa is a notorious pathogen that causes biofilm aided infections in patients with cystic fibrosis and burn wounds, resulting in significant mortality in immunocompromised individuals. This study reports a novel one-step biosynthesis of gold nanoparticles using phytocompound, hordenine (HD), as a reducing and capping agent. The synthesis of the anisotropic hordenine-fabricated gold nanoparticles (HD-AuNPs) with an average particle size of 136.87 nm was achieved within 12 h of incubation at room temperature...
December 2017: IET Nanobiotechnology
https://www.readbyqxmd.com/read/29155139/the-generation-and-characterisation-of-neutralising-antibodies-against-the-theiler-s-murine-encephalomyelitis-virus-tmev-gdvii-capsid-reveals-the-potential-binding-site-of-the-host-cell-co-receptor-heparan-sulfate
#7
Nicole Upfold, Caroline Ross, Özlem Tastan Bishop, Garry A Luke, Caroline Knox
The early stages of picornavirus capsid assembly and the host factors involved are poorly understood. Since the localisation of viral proteins in infected cells can provide information on their function, antibodies against purified Theiler's murine encephalomyelitis virus (TMEV) GDVII capsids were generated by immunisation of rabbits. The resultant anti-TMEV capsid antibodies recognised a C-terminal region of VP1 but not VP2 or VP3 by Western analysis. Examination of the sites of TMEV capsid assembly by indirect immunofluorescence and confocal microscopy showed that at 5hours post infection, capsid signal was diffusely cytoplasmic with strong perinuclear staining and moved into large punctate structures from 6 to 8hours post infection...
November 16, 2017: Virus Research
https://www.readbyqxmd.com/read/29155097/14-3-3z-sequesters-cytosolic-t-bet-up-regulating-il-13-in-tc2-and-scleroderma-cd8-lymphocytes
#8
Sandra Cascio, Thomas A Medsger, William F Hawse, Simon C Watkins, Christine Milcarek, Larry W Moreland, Robert A Lafyatis, Patrizia Fuschiotti
BACKGROUND: Interleukin(IL)-13-producing CD8+ T cells have been implicated in the pathogenesis of type-2 driven inflammatory human conditions. We have shown that CD8+IL-13+ cells play a critical role in cutaneous fibrosis, the most characteristic feature of systemic sclerosis (scleroderma; SSc). However, the molecular mechanisms underlying IL-13 and other type-2 cytokine production by CD8+ T cells remain unclear. OBJECTIVE: Establish the molecular basis of IL-13 over-production by SSc CD8+ T cells, focusing on T-bet modulation of GATA-3 activity, which we showed to underlie IL-13 over-production in SSc CD8+IL-13+ cells...
November 15, 2017: Journal of Allergy and Clinical Immunology
https://www.readbyqxmd.com/read/29153956/effect-of-laser-activated-bleaching-on-the-chemical-stability-and-morphology-of-intracoronal-dentin
#9
Fabiane Carneiro Lopes, Renato Roperto, Anna Akkus, Ozan Akkus, Regina Guenka Palma-Dibb, Manoel Damião de Sousa-Neto
OBJECTIVES: To evaluate the effect of the bleaching with 35% hydrogen peroxide either activated or not by a 970nm diode laser on the chemical stability and dentin surface morphology of intracoronary dentin. METHODS: Twenty-seven slabs of intracoronary dentin specimens (3×3mm) were distributed into three groups (n=9), according to surface treatment: HP - 35% hydrogen peroxide (1×4'), DL - 970nm diode laser (1×30"/0,8W/10Hz), HP+DL - 35% HP activated with 970nm diode laser (1×30"/0,8W/10Hz leaving the gel in contact to the surface for 4' after activation)...
November 13, 2017: Archives of Oral Biology
https://www.readbyqxmd.com/read/29153715/cisplatin-and-doxorubicin-high-loaded-nanodrug-based-on-biocompatible-thioether-and-ethane-bridged-hollow-mesoporous-organosilica-nanoparticles
#10
Junjie Zhang, Lixing Weng, Xiaodan Su, Guangming Lu, Wenfei Liu, Yuxia Tang, Yunlei Zhang, Jun Wen, Zhaogang Teng, Lianhui Wang
Herein, a mesoporous organosilica nanoparticle (MON) based nanodrug highly loaded with cisplatin (CDDP) and doxorubicin (DOX) (denoted as MONs/CDDP/DOX) has been successfully prepared for the first time. The MONs are characterized with core-contained double hollow shells, thioether and ethane groups separately incorporated frameworks, uniform diameter (420 nm), large surface area (592 m(2)/g), and ordered pore size (2.5 nm). The safety evaluation of the MONs based on cell viability, haemolytic activity, histological change, and serum biochemical index demonstrates that they have excellent biological compatibility...
November 2, 2017: Journal of Colloid and Interface Science
https://www.readbyqxmd.com/read/29152726/autoantibodies-to-full-body-vascular-cell-junctions-colocalize-with-myzap-arvcf-desmoplakins-i-and-ii-and-p0071-in-endemic-pemphigus-in-colombia-south-america
#11
Ana M Abreu Velez, Hong Yi, Gunnar Warfvinge, Michael S Howard
BACKGROUND: We previously described a new variant of endemic pemphigus foliaceus in El Bagre, Colombia (El Bagre-EPF). METHODS: Here we aimed to investigate disease autoreactivity to vessels in all body organs/systems. We compared 57 patients and 57 controls from the endemic area, matched by demographics, age, sex, and work activity. We performed immunofluorescence, immunohistochemistry, confocal microscopy, immunoblotting, indirect immune electron microscopy studies, and autometallographic studies...
November 20, 2017: International Journal of Dermatology
https://www.readbyqxmd.com/read/29152606/ethanol-induced-steatosis-involves-impairment-of-lipophagy-associated-with-reduced-dynamin2-activity
#12
Karuna Rasineni, Terrence M Donohue, Paul G Thomes, Li Yang, Dean J Tuma, Mark A McNiven, Carol A Casey
Background: Lipid droplets (LDs), the organelles central to alcoholic steatosis, are broken down by lipophagy, a specialized form of autophagy. Here, we hypothesize that ethanol administration retards lipophagy by down-regulating Dynamin 2 (Dyn2), a protein that facilitates lysosome re-formation, contributing to hepatocellular steatosis. Methods: Primary hepatocytes were isolated from male Wistar rats fed Lieber-DeCarli control or EtOH liquid diets for 6-8 wk. Hepatocytes were incubated in complete medium (fed) or nutrient-free medium (fasting) with or without the Dyn2 inhibitor Dynasore or the Src inhibitor SU6656...
August 2017: Hepatol Commun
https://www.readbyqxmd.com/read/29152089/high-glucose-downregulates-myocardin-expression-in-rat-glomerular-mesangial-cells-via-the-erk-signaling-pathway
#13
Ming Li, Lijuan Xu, Guowei Feng, Yan Zhang, Xin Wang, Yuebing Wang
Mesangial cells (MCs), which are vascular smooth muscle-derived cells, occupy the central position in the glomerulus. Diabetic nephropathy (DN) is one of the most common diabetes complications and is likely attributed to the loss of MC contractility. Myocardin stimulates downstream vascular smooth muscle genes and regulates the contractility of vascular smooth muscle cells. Therefore, we hypothesized that myocardin is expressed in MCs and that high glucose is involved in the regulation of myocardin and downstream contractile genes in the context of DN...
October 20, 2017: Oncotarget
https://www.readbyqxmd.com/read/29151647/7-10-epoxyoctadeca-7-9-dienoic-acid-a-small-molecule-adjuvant-that-potentiates-%C3%AE-lactam-antibiotics-against-multidrug-resistant-staphylococcus-aureus
#14
Chakradhar Dasagrandhi, Young-Soon Kim, In-Hwan Kim, Ching T Hou, Hak-Ryul Kim
The emergence of methicillin-resistant Staphylococcus aureus (MRSA) infections with multi-drug resistance needs effective and alternative control strategies. In this study we investigated the adjuvant effect of a novel furan fatty acid, 7,10-epoxyoctadeca-7,9-dienoic acid (7,10-EODA) against multidrug-resistant S. aureus (MDRSA) strain 01ST001 by disc diffusion, checker board and time kill assays. Further the membrane targeting action of 7,10-EODA was investigated by spectroscopic and confocal microscopic studies...
December 2017: Indian Journal of Microbiology
https://www.readbyqxmd.com/read/29151350/tracking-of-fluorescently-labeled-polymer-particles-reveals-surface-effects-during-shear-controlled-aggregation
#15
Stefano Caimi, Alberto Cingolani, Baptiste Jaquet, Marc Siggel, Marco Lattuada, Massimo Morbidelli
Surface chemistry is believed to be the key parameter affecting the aggregation and breakage of colloidal suspensions when subjected to shear. To date, only a few works dealt with the understanding of the role of the physical and chemical properties of the particles' surface upon aggregation under shear. Previous studies suggested that surface modifications strongly affect polymer particles' adhesion, but it resulted very challenging to demonstrate this effect and monitor these alterations upon prolonged exposure to shear forces...
November 20, 2017: Langmuir: the ACS Journal of Surfaces and Colloids
https://www.readbyqxmd.com/read/29150331/anti-amyloidogenic-and-anti-apoptotic-effect-of-%C3%AE-bisabolol-against-a%C3%AE-induced-neurotoxicity-in-pc12-cells
#16
Balakrishnan Shanmuganathan, Venkatesan Suryanarayanan, Sethuraman Sathya, Muralidharan Narenkumar, Sanjeev Kumar Singh, Kandasamy Ruckmani, Kasi Pandima Devi
Alzheimer's disease (AD) is a life-threatening neurodegenerative disorder leading to dementia, with a progressive decline in memory and other thinking skills of elderly populace. Of the multiple etiological factors of AD, the accumulation of senile plaques (SPs) particularly as Aβ oligomers correlates with the relentlessness cognitive impairment in AD patients and play a vital role in AD pathology. Since natural essential oil constituents have successfully served as a source of drugs for AD treatment, the present study aims at the in vitro and in silico investigation of anti-amyloidogenic potential and anti-apoptotic property of the α-bisabolol against Aβ25-35 induced neurotoxicity in PC12 cells...
October 10, 2017: European Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/29149542/spectrally-encoded-coherence-tomography-and-reflectometry-sectr-simultaneous-en-face-and-cross-sectional-imaging-at-2-gigapixels-per-second
#17
Mohamed T El-Haddad, Ivan Bozic, Yuankai K Tao
Non-invasive biological imaging is crucial for understanding in vivo structure and function. Optical coherence tomography (OCT) and reflectance confocal microscopy are two of the most widely used optical modalities for exogenous contrast-free high-resolution three-dimensional imaging in non-fluorescent scattering tissues. However, sample motion remains a critical barrier to raster-scanned acquisition and reconstruction of wide-field anatomically accurate volumetric datasets. We introduce spectrally encoded coherence tomography and reflectometry (SECTR), a high-speed multimodality system for simultaneous OCT and spectrally-encoded reflectance (SER) imaging...
November 17, 2017: Journal of Biophotonics
https://www.readbyqxmd.com/read/29149370/-hematological-diagnosis-in-the-corneal-consultation
#18
J Wasielica-Poslednik, A Gericke, M Munder, N Pfeiffer, W Lisch
This case report describes a diagnostic chain in a patient with atypical bilateral corneal opacity, which led to the diagnosis of a hematological disorder. The patient's medical history, clinical appearance and findings in confocal microscopy gave rise to the suspicion of a paraproteinemic keratopathy. The hematological laboratory diagnostics revealed a monoclonal gammopathy of the IgG kappa type. The bone marrow puncture led to the diagnosis of a lymphoplasmacytic lymphoma, which belongs to the group of B‑cell non-Hodgkin's lymphomas...
November 17, 2017: Der Ophthalmologe: Zeitschrift der Deutschen Ophthalmologischen Gesellschaft
https://www.readbyqxmd.com/read/29148780/a-spectroscopic-study-of-the-interaction-of-carboxyl-modified-gold-nanoparticles-with-liposomes-of-different-chain-lengths-and-controlled-drug-release-by-layer-by-layer-technology
#19
Nishu Kanwa, Soumya Kanti Dey, Chandan Adhikari, Anjan Chakraborty
In this manuscript, we investigate the interactions of carboxyl modified gold nanoparticles (AuC) with zwitterionic phospholipid liposomes of different chain lengths using a well-known membrane probe PRODAN by steady state and time resolved spectroscopy. We used three zwitterionic lipids, namely, dipalmitoylphosphatidylcholine (DPPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), and 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC); which are widely different in their phase transition temperatures to form liposome-AuC assemblies...
November 17, 2017: Journal of Physical Chemistry. B
https://www.readbyqxmd.com/read/29148709/nir-emitting-nanoprobes-based-on-cyclic-rgd-motif-conjugated-pbs-quantum-dots-for-integrin-targeted-optical-bioimaging
#20
Nicoletta Depalo, Michela Corricelli, Ivan de Paola, Gianpiero Valente, Rosa M Iacobazzi, Emiliano Altamura, Doriana Debellis, Daniela Comegna, Elisabetta Fanizza, Nunzio Denora, Valentino Laquintana, Fabio Mavelli, Marinella Striccoli, Michele Saviano, Angela Agostiano, Annarita Del Gatto, Laura Zaccaro, Maria Lucia Curri
Here, silica coated PbS quantum dots (QDs) with photoluminesce emission properties in the near-infrared (NIR) region are proposed as potential effective single particle optical nanoprobes for future in vivo imaging of tumors. The dispersibility in aqueous medium of hydrophobic PbS QDs was accomplished by growing a silica shell on their surface by exploiting a base assisted water-in-oil microemulsion method. The silica coated PbS QDs were then conjugated with specifically designed cyclic arginine-glycine-aspartic acid (cRGD) peptide, that are able to specifically recognizes the αvβ3 integrins which is overexpressed in the angiogenic tumor induced vasculatures and on some solid tumours, in order to achieve a tumor-specific targeting...
November 17, 2017: ACS Applied Materials & Interfaces
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