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confocal and microscopy

Lélia Abad, Virginie Tafani, Jason Tasse, Jérôme Josse, Christian Chidiac, Sébastien Lustig, Tristan Ferry, Alan Diot, Frédéric Laurent, Florent Valour
Objectives: Prolonged use of linezolid for bone and joint infection (BJI) is limited by its long-term toxicity. The better safety profile of tedizolid, a recently developed oxazolidinone, could offer an alternative. However, its efficacy against biofilm-embedded and intracellular Staphylococcus aureus, the two main bacterial reservoirs associated with BJI chronicity, is unknown. Methods: Using three S. aureus strains (6850 and two clinical BJI isolates), linezolid and tedizolid were compared regarding their ability: (i) to target the S...
December 4, 2018: Journal of Antimicrobial Chemotherapy
Jianhua Liu, Weilin Liu, Louise J Salt, Mike J Ridout, Yuting Ding, Peter J Wilde
Incorporation of fish oil containing ω-3 polyunsaturated fatty acids (PUFAs) into functional foods remains challenging. In this study, caseinate and glycoconjugates (CD6, CD40, CD70, CD100) of caseinate to dextrans of different molecular weights (D6, D40, D70, D100 kDa) were used to stabilize fish oil emulsions, and the impact on physicochemical stability and gastrointestinal fate was investigated. The glycoconjugate of CD6 exhibited significantly higher conjugation efficiency, lower surface hydrophobicity (H0) and lower surface activity than other glycoconjugates...
December 5, 2018: Journal of Agricultural and Food Chemistry
Cuiping Jiang, Zitong Qi, Hengbo Jia, Yilei Huang, Yunbo Wang, Wenli Zhang, Zimei Wu, Hu Yang, Jianping Liu
In this work, we report on an ATP-responsive low-molecular-weight polyethylenimine (LMW-PEI)-based supramolecular assembly. It formed via host-guest interaction between PEI (MW=1.8 kDa)-α-cyclodextrin (α-CD) conjugates and PEI1.8k-phenylboronic acid (PBA) conjugates. The host-guest interaction between PEI1.8k-α-CD and PEI1.8k-PBA was confirmed by the 2D-NOESY chromatogram experiment and competition test. The ATP-responsive property of the supramolecular assembly was evaluated by a series of ATP-triggered degradation and siRNA release studies in terms of fluorescence resonance energy transfer, agarose gel electrophoresis assay and the time course monitoring of the particle size and morphology...
December 5, 2018: Biomacromolecules
Hidetaka Miyagi, Soohyun Kim, Jennifer Li, Christopher J Murphy, Sara M Thomasy
PURPOSE: Corneal endothelial cell regeneration varies by species, with nonhuman primates (NHPs) and rabbits displaying low and high proliferative capacities, respectively. Recent studies report that topical application of rho-associated kinase (ROCK) inhibitors accelerates corneal endothelial wound healing in animal models and human patients with endothelial dysfunction. This study determines the regenerative capacity of canine corneal endothelial cells in vivo and their response to a topical ROCK inhibitor, Y27632, after transcorneal freezing...
November 29, 2018: Cornea
Congshan Liu, Jiaqing Yao, Jianhai Yin, Jian Xue, Haobing Zhang
Echinococcosis, which causes a high disease burden and is of great public health significance, is caused by the larval stage of Echinococcus species. It has been suggested that tubulin is the target of benzimidazoles, the only drugs for the treatment of echinococcosis. This study evaluated the characteristics of tubulins from Echinococcus granulosus. The full-length cDNAs of E. granulosus α- and β-tubulin isoforms were cloned by reverse transcription PCR from protoscolex RNA. Then, these two tubulin isoforms (α9 and β4 ) were recombinantly expressed as insoluble inclusion bodies in Escherichia coli...
2018: Parasite: Journal de la Société Française de Parasitologie
Nida Mubin, Mohd Saad Umar, Swaleha Zubair, Mohammad Owais
In the present study, we investigated potential of chitosan-based nanoparticles (CNPs) to deliver loaded therapeutic molecules to pathogen harboring macrophages. We fabricated stable CNPs employing ionic cross-linking method and evaluated their potential to target RAW 264.7 cells. The physicochemical characterization of as-synthesized CNPs was determined using electron microscopy, infrared microscopy and zeta potential measurement. Next, cellular uptake and intracellular localization studies of CNPs were followed in living RAW264...
2018: Frontiers in Microbiology
Inmaculada Mora-Espí, Lleonard Barrios, Elena Ibáñez, Jorge Soriano, Carme Nogués
Therapeutic drug carriers can drive their cargo to their target cells. However, an obstacle is usually the entrapment of the drug inside the endolysosomal compartment, which physically impedes its actuation by the impossibility of reaching its molecular site of action. To overcome this hurdle, photochemical internalization (PCI) has been proposed, but the extent of PCI-induced membrane disruption and its capability to allow the release of microparticles is unknown. The aim of the present study was to determine if PCI allows the release of microparticles from the endolysosomal compartment to the cytosol and to analyze at the ultrastructural level the effect of PCI on the membrane surrounding the particles...
December 4, 2018: Scientific Reports
Emanuela Roscetto, Patrizia Contursi, Adriana Vollaro, Salvatore Fusco, Eugenio Notomista, Maria Rosaria Catania
Candida species cause cutaneous and systemic infections with a high mortality rate, especially in immunocompromised patients. The emergence of resistance to the most common antifungal drugs, also due to biofilm formation, requires the development of alternative antifungal agents. The antimicrobial peptide VLL-28, isolated from an archaeal transcription factor, shows comparable antifungal activity against 10 clinical isolates of Candida spp. Using a fluoresceinated derivative of this peptide, we found that VLL-28 binds to the surface of planktonic cells...
December 4, 2018: Scientific Reports
Frederike M Muskens, Richard J Ward, Dominik Herkt, Helmus van de Langemheen, Andrew B Tobin, Rob M J Liskamp, Graeme Milligan
Chemoproteomic approaches to identify ligand-receptor interactions have gained popularity however, identifying trans-membrane receptors remains challenging. A new tri-functional probe to aid the non-biased identification of such receptors was developed and synthesized using a convenient 7-step synthesis. This probe contained three functional groups: (1) an NHS-ester for ligand-coupling through free amines; (2) a diazirine moiety to capture the receptor of interest upon irradiation with UV-light; and (3) a biotin group which allowed affinity purification of the final adduct using streptavidin...
December 4, 2018: Molecular Pharmacology
Xuechao Han, Jingman Xu, Sen Xu, Yahan Sun, Mali He, Xiaodong Li, Xinyu Li, Jiayi Pi, Rui Yu, Wei Tian
OBJECTIVE: To explore the role of mitochondrial permeability transition pore (mPTP) in mediating the protective effect of gastrodin against oxidative stress damage in H9c2 cardiac myocytes. METHODS: H9c2 cardiac myocytes were treated with H2 O2 , gastrodin, gastrodin+H2 O2 , cyclosporin A (CsA), or CsA+gas+H2 O2 group. MTT assay was used to detect the survival ratio of H9c2 cells, and flow cytometry with Annexin V-FITC/PI double staining was used to analyze the early apoptosis rate after the treatments...
November 30, 2018: Nan Fang Yi Ke da Xue Xue Bao, Journal of Southern Medical University
Olivia L Lee, Tudor C Tepelus, Jianyan Huang, Anne G Irvine, Christopher Irvine, Gloria B Chiu, SriniVas R Sadda
BACKGROUND: The corneal epithelium is directly affected in dry eye syndrome. Thus, we attempted to describe the morphological features and evaluate the cellular density within the corneal epithelial layers in patients with non-Sjögren's (NSDE) and Sjögren's syndrome dry eyes (SSDE) by in vivo confocal microscopy (IVCM). METHODS: Central cornea was prospectively imaged by IVCM in 68 clinically diagnosed aqueous tear-deficient dry eyes and 10 healthy age-matched control eyes...
December 4, 2018: BMC Ophthalmology
Carol Korzeniewski, Jay P Kitt, Saheed Bukola, Stephen E Creager, Shelley D Minteer, Joel M Harris
Single layer graphene (SLG), with its angstrom-scale thickness and strong Raman scattering cross section, was adapted for measurement of the axial (Z-direction) probe beam profile in confocal Raman microscopy depth-profiling experiments. SLG adsorbed to a glass microscope coverslip (SLG/SiO2 ) served as a platform for estimation of axial spatial resolution. Profiles were measured by stepping the confocal probe volume through the SLG/SiO2 interface while measuring Raman scattering from the sample. Using a high numerical aperture (1...
December 4, 2018: Analytical Chemistry
C Longo, R Pampena, C Bombonato, S Gardini, S Piana, M Mirra, M Raucci, A Kyrgidis, G Pellacani, M Ragazzi
BACKGROUND: Frozen histological sections are currently used for intra-operative margin assessment during Mohs surgery. Fluorescence confocal microscopy (FCM) is a new tool, which offers a promising and faster alternative to frozen histology. OBJECTIVE: The aim of the present study is to prospectively evaluate in a clinical setting, the accuracy of FCM as compared to frozen sections in BCC's margin assessment. METHODS: Patients with BCC, scheduled for Mohs surgery were prospectively enrolled...
December 4, 2018: British Journal of Dermatology
Gil-Hyun Kim, Hang-Gu Kim, Chang-Jin Jeon
The purpose of the present study was to investigate the organization of choline acetyltransferase (ChAT)-immunoreactive (IR) fibers in the visual cortex of the microbat, using standard immunocytochemistry and confocal microscopy. ChAT-IR fibers were distributed throughout all layers of the visual cortex, with the highest density in layer III and the lowest density in layer I. However, no ChAT-IR cells were found in the microbat visual cortex. ChAT-IR fibers were classified into two types: small and large varicose fibers...
October 31, 2018: Acta Histochemica et Cytochemica
Christian Møller-Olsen, Siu Fung Stanley Ho, Ranti Dev Shukla, Tamas Feher, Antonia P Sagona
Bacterial infections can be treated with bacteriophages that show great specificity towards their bacterial host and can be genetically modified for different applications. However, whether and how bacteriophages can kill intracellular bacteria in human cells remains elusive. Here, using CRISPR/Cas selection, we have engineered a fluorescent bacteriophage specific for E. coli K1, a nosocomial pathogen responsible for urinary tract infections, neonatal meningitis and sepsis. By confocal and live microscopy, we show that engineered bacteriophages K1F-GFP and E...
December 3, 2018: Scientific Reports
Dmitri I Kotov, Thomas Pengo, Jason S Mitchell, Matthew J Gastinger, Marc K Jenkins
Advances in imaging have led to the development of powerful multispectral, quantitative imaging techniques, like histo-cytometry. The utility of this approach is limited, however, by the need for time consuming manual image analysis. We therefore developed the software Chrysalis and a group of Imaris Xtensions to automate this process. The resulting automation allowed for high-throughput histo-cytometry analysis of three-dimensional confocal microscopy and two-photon time-lapse images of T cell-dendritic cell interactions in mouse spleens...
December 3, 2018: Journal of Immunology: Official Journal of the American Association of Immunologists
Zhenhua Chen, Guanyu Chen, Kang Zhou, Peng Zhang, Xiuli Ren, Xifan Mei
In this paper, the effect of commonly used food sweetener (sodium cyclamate) on the proliferation and differentiation of osteoblasts has been researched. The morophology change of osteoblasts was investigated by confocal laser scanning microscopy. Cell viability was studied by MTT analysis. BMP2 expression was analyzed by western blot and immunofluorescence. Mineralization ability of osteoblasts was researched by using alizarin red staining method. The results indicate that a very low concentration (0.06 μM) of sodium cyclamate can curle and fold microfilament and microtubule of osteoblasts...
November 30, 2018: Biochemical and Biophysical Research Communications
U Daood, C K Y Yiu
OBJECTIVE: To evaluate the transdentinal cytotoxicity and macrophage phenotype response to a novel quaternary ammonium silane (QAS) cavity disinfectant. METHODS: NIH 3T3 mouse fibroblasts were cultured in Dulbecco's Modified Eagle's Medium and incubated for 3 days. The cells (3×104 ) were seeded on the pulpal side of dentine discs and the occlusal side of the discs were treated with different cavity disinfectants: Group 1: de-ionized water (control); Group 2: 2% chlorhexidine (CHX); Group 3: 2% QAS; Group 4: 5% QAS, and Group 5: 10% QAS...
November 30, 2018: Dental Materials: Official Publication of the Academy of Dental Materials
Petra Švančarová, Tatiana Betáková
BACKGROUND: The influenza matrix protein (M1) layer under the viral membrane plays multiple roles in virus assembly and infection. N-domain and C-domain are connected by a loop region, which consists of conserved RQMV motif. METHODS: The function of the highly conserve RQMV motif in the influenza virus life cycle was investigated by site-directed mutagenesis and by rescuing mutant viruses by reverse genetics. Co-localization of M1 with nucleoprotein (NP), clustered mitochondria homolog protein (CLUH), chromosome region maintenance 1 protein (CRM1), or plasma membrane were studied by confocal microscopy...
December 3, 2018: Virology Journal
Hye-Yeong Kim, Ran Li, Thomas S C Ng, Gabriel Courties, Christopher Blake Rodell, Mark Prytyskach, Rainer H Kohler, Mikael Pittet, Matthias Nahrendorf, Ralph Weissleder, Miles A Miller
Tumor associated macrophages (TAMs) are widely implicated in cancer progression, and TAM levels can influence drug responses, particularly to immunotherapy and nanomedicines. However, it has been difficult to quantify total TAM numbers and their dynamic spatiotemporal distribution in a non-invasive and translationally relevant manner. Here, we address this need by developing a pharmacokinetically optimized, 64Cu-labeled polyglucose nanoparticle (Macrin) for quantitative positron emission tomography (PET) imaging of macrophages in tumors...
December 3, 2018: ACS Nano
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