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confocal microscopy

Gerardo Pedro García-García, Magdalena Martínez-Rubio, Medina-Azahara Moya-Moya, Juan José Pérez-Santonja, Julio Escribano
PURPOSE: To identify the spectrum of disease-causing CYP4V2 variants in Spanish patients with clinically diagnosed Bietti crystalline dystrophy (BCD) over an 8-year period and to analyse the phenotype-genotype correlation of the identified variants. METHODS: Four unrelated Spanish probands with a clinical diagnosis of BCD were recruited. Ophthalmological examination included visual acuity (VA), slit lamp examination, in vivo corneal confocal microscopy, funduscopy and fluoresceinic angiography...
April 25, 2018: Acta Ophthalmologica
Cihan Kayasandik, Pooran Negi, Fernanda Laezza, Manos Papadakis, Demetrio Labate
Fluorescence confocal microscopy has become increasingly more important in neuroscience due to its applications in image-based screening and profiling of neurons. Multispectral confocal imaging is useful to simultaneously probe for distribution of multiple analytes over networks of neurons. However, current automated image analysis algorithms are not designed to extract single-neuron arbors in images where neurons are not separated, hampering the ability map fluorescence signals at the single cell level. To overcome this limitation, we introduce NeuroTreeTracer - a novel image processing framework aimed at automatically extracting and sorting single-neuron traces in fluorescent images of multicellular neuronal networks...
April 24, 2018: Scientific Reports
Julie Gaburro, Asim Bhatti, Jenni Harper, Isabelle Jeanne, Megan Dearnley, Diane Green, Saeid Nahavandi, Prasad N Paradkar, Jean-Bernard Duchemin
Understanding Zika virus infection dynamics is essential, as its recent emergence revealed possible devastating neuropathologies in humans, thus causing a major threat to public health worldwide. Recent research allowed breakthrough in our understanding of the virus and host pathogenesis; however, little is known on its impact on its main vector, Aedes aegypti. Here we show how Zika virus targets Aedes aegypti's neurons and induces changes in its behavior. Results are compared to dengue virus, another flavivirus, which triggers a different pattern of behavioral changes...
April 25, 2018: Emerging Microbes & Infections
Joanna Klubo-Gwiezdzinska, Shilpa Thakur, Brianna Daley, Kelli Gaskins, Vasyl V Vasko, Myriem Boufraqech, Dhaval Patel, Carole Sourbier, Jeff M Reece, Sheue-Yann Cheng, Electron Kebebew, Sunita K Agarwal
PURPOSE: Mitochondrial glycerophosphate dehydrogenase (mGPDH) is the key enzyme connecting oxidative phosphorylation (OXPHOS) and glycolysis as well as a target of the antidiabetic drug metformin (MF) in the liver. There are no data on the expression and role of mGPDH as a metformin target in cancer. In this study, we evaluated mGPDH as a potential target of metformin in thyroid cancer and investigated its contribution in thyroid cancer metabolism. EXPERIMENTAL DESIGN: We analyzed mGPDH expression in 253 thyroid cancer and normal tissues by immunostaining and examined its expression and localization in thyroid cancer-derived cell lines (FTC133, BCPAP) by confocal microscopy...
April 24, 2018: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
Hechao Du, Jie Yang, Zhaoxin Lu, Xiaomei Bie, Haizhen Zhao, Chong Zhang, Fengxia Lu
Bacillus cereus is an opportunistic human pathogen that causes serious foodborne diseases and need to be controlled using natural preservatives. We isolated and elucidated the mode of action of plantaricin GZ1-27, a novel bacteriocin that potently inhibits B. cereus. Plantaricin GZ1-27 was purified using ammonium sulfate precipitation, gel-filtration chromatography, and reverse-phase HPLC, and Q-TOF/MS analysis revealed that its molecular mass was 975 Da. Plantaricin GZ1-27 was highly thermostable, pH and organic-solvent tolerant, and trypsin and pepsin sensitive...
April 25, 2018: Journal of Agricultural and Food Chemistry
Pakorn Kraisit, Narong Sarisuta
The aim of this present work was to prepare triamcinolone acetonide (TA)-loaded nanostructured lipid carriers (TA-loaded NLCs) for buccal drug delivery systems using the Box-Behnken design. A hot homogenization method was used to prepare the TA-loaded NLCs. Spermaceti (X₁), soybean oil (X₂), and Tween 80 (X₃) were used as solid lipid, liquid lipid, and stabilizer, respectively. The particle size of TA-loaded NLCs was lower than 200 nm and the zeta potential displayed the negative charge in all formulations...
April 23, 2018: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
Manuela Leri, Matteo Ramazzotti, Marzia Vasarri, Sara Peri, Emanuela Barletta, Carlo Pretti, Donatella Degl'Innocenti
Posidonia oceanica (L.) Delile is a marine plant with interesting biological properties potentially ascribed to the synergistic combination of bioactive compounds. Our previously described extract, obtained from the leaves of P. oceanica , showed the ability to impair HT1080 cell migration by targeting both expression and activity of gelatinases. Commonly, the lack of knowledge about the mechanism of action of phytocomplexes may be an obstacle regarding their therapeutic use and development. The aim of this study was to gain insight into the molecular signaling through which such bioactive compounds impact on malignant cell migration and gelatinolytic activity...
April 21, 2018: Marine Drugs
Mariana E Elizondo-García, Valeria Márquez-Miranda, Ingrid Araya-Durán, Jesús A Valencia-Gallegos, Fernando D González-Nilo
Amphiphilic Janus dendrimers (JDs) are repetitively branched molecules with hydrophilic and hydrophobic components that self-assemble in water to form a variety of morphologies, including vesicles analogous to liposomes with potential pharmaceutical and medical application. To date, the self-assembly of JDs has not been fully investigated thus it is important to gain insight into its mechanism and dependence on JDs’ molecular structure. In this study, the aggregation behavior in water of a second-generation bis-MPA JD was evaluated using experimental and computational methods...
April 21, 2018: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
L C Gomes, J Deschamps, R Briandet, F J Mergulhão
This work investigated the effects of diamond-like carbon (DLC) coatings on the architecture and biocide reactivity of dual-species biofilms mimicking food processing contaminants. Biofilms were grown using industrial isolates of Escherichia coli and Pantoea agglomerans on bare stainless steel (SST) and on two DLC surface coatings (a-C:H:Si:O designated by SICON® and a-C:H:Si designated by SICAN) in order to evaluate their antifouling activities. Quantification and spatial organization in single- and dual-species biofilms were examined by confocal laser scanning microscopy (CLSM) using a strain specific labelling procedure...
April 12, 2018: International Journal of Food Microbiology
P D'Incecco, L Ong, S Gras, L Pellegrino
Non-pathogenic spore-forming Clostridia are of increasing interest due to their application in biogas production and their capability to spoil different food products. The life cycle for Clostridium includes a spore stage that can assist in survival under environmentally stressful conditions, such as extremes of temperature or pH. Due to their size, spores can be investigated by a range of microscopic techniques, many of which involve sample pre-treatment. We have developed a quick, simple and non-destructive fluorescent staining procedure that allows a clear differentiation between spores and vegetative cells and effectively stains spores, allowing recovery and tracking in subsequent experiments...
April 18, 2018: Micron: the International Research and Review Journal for Microscopy
Nan Jiang, Hyeon-Jin Kim, Tyler J Chozinski, Jorge E Azpurua, Benjamin A Eaton, Joshua C Vaughan, Jay Z Parrish
The limited resolving power of conventional diffraction-limited microscopy hinders analysis of small, densely packed structural elements in cells. Expansion Microscopy (ExM) provides an elegant solution to this problem, allowing for increased resolution with standard microscopes via physical expansion of the specimen in a swellable polymer hydrogel. Here, we apply, validate, and optimize ExM protocols that enable the study of Drosophila embryos, larval brains, and larval and adult body walls. We achieve a lateral resolution of ∼70 nm in Drosophila tissues using a standard confocal microscope, and we use ExM to analyze fine intracellular structures and intercellular interactions...
April 24, 2018: Molecular Biology of the Cell
Shengyu Zhang, Yanjun Liu, Ye Gan, Nanqing Qiu, Yueqing Gu, Hongyan Zhu
The key for better anti-tumor efficacy is to improve the specificity of anti-tumor drugs for tumor cells and diminish their cytotoxicity to normal tissues. Targeted nanoparticles as anti-tumor drug delivery system can resolve this problem. In this study, we prepared folate and TAT (arginine-rich cell penetrating peptide) modified N-PEG-N'-octyl-chitosan to form the folate/TAT-PEG-OC micelles. Then the molecular structure, morphology, size distribution and bio-safety of the micelles were characterized. In order to investigate the drug loading capacity of folate/TAT-PEG-OC micelles, doxorubicin (DOX) was used as model drug to prepare DOX-loaded chitosan micelles...
April 24, 2018: Pharmaceutical Development and Technology
Roghayeh Rastegar, Hamid Akbari Javar, Mehdi Khoobi, Poua Dehghan Kelishadi, Gholam Hossein Yousefi, Mahmoud Doosti, Mohammad Hossien Ghahremani, Ahmad Sharif Tabrizi, Fatemeh Imanparast, Elham Gholibeglu, Mahdi Gholami
Codelivery of chemo-sensitizers with chemotherapeutics using combo nanomedicine is a promising platform for overcoming chemoresistance in breast cancer. However, tumor accumulation of nano-carriers based on enhanced permeability and retention (EPR) effect is confounded by heterogeneity in tumor microenvironment. Adsorption of protein corona on surface of nanoparticle boost up clearance by reticulo-endothelial system. In this study, a surface functionalized magnetic nanocomposite (NC) for codelivery of doxorubicin (DOX) and curcumin (CUR) is developed...
April 24, 2018: Artificial Cells, Nanomedicine, and Biotechnology
E Benati, R Pampena, C Bombonato, S Borsari, M Lombardi, C Longo
Actinic cheilitis (AC) can precede the development of squamous cell carcinoma (SCC) of the lip, a location with higher risk of invasiveness and metastasis. Herein, we reported the use of ingenol mebutate (IngMeb) 0.015% gel on three consecutive days to treat three patients suffering from AC. All the three patients achieved complete clearance of AC with rapid clinical effect, favorable safety profile, good patient's compliance related to short time of applications, and few local skin reactions. So IngMeb is an attractive new therapy for AC...
April 24, 2018: Dermatologic Therapy
Shuai Ye, Wei Yan, Mengjie Zhao, Xiao Peng, Jun Song, Junle Qu
Stimulated emission depletion (STED) nanoscopy is one of the most promising super-resolution imaging techniques for microstructure imaging. Commercial CdSe@ZnS quantum dots are used as STED probes and ≈50 nm lateral resolution is obtained. Compared with other quantum dots, perovskite CsPbBr3 nanoparticles (NPs) possess higher photoluminescence quantum yield and larger absorption cross-section, making them a more effective probe for STED nanoscopy. In this study, CsPbBr3 NPs are used as probes for STED nanoscopy imaging...
April 24, 2018: Advanced Materials
Dominique Mayrand, Julie Fradette
Optimal imaging methods are necessary in order to perform a detailed characterization of thick tissue samples from either native or engineered tissues. Tissue-engineered substitutes are featuring increasing complexity including multiple cell types and capillary-like networks. Therefore, technical approaches allowing the visualization of the inner structural organization and cellular composition of tissues are needed. This chapter describes an optical clearing technique which facilitates the detailed characterization of whole-mount samples from skin and adipose tissues (ex vivo tissues and in vitro tissue-engineered substitutes) when combined with spectral confocal microscopy and quantitative analysis on image renderings...
2018: Methods in Molecular Biology
Gloria Roberti, Luca Agnifili, Francesca Berardo, Ivano Riva, Michele Figus, Gianluca Manni, Luciano Quaranta, Francesco Oddone
INTRODUCTION: To compare the effects of a preservative-free (PF) ophthalmic solution containing hyaluronic acid (HA) 0.4% and taurine (TAU) 0.5% with those of a PF ophthalmic solution containing HA 0.2% on ocular surface signs, symptoms, and morphological parameters in glaucoma patients under multiple long-term topical hypotensive therapy. METHODS: Eligible patients underwent evaluation of ocular surface parameters by ocular surface disease index (OSDI) and glaucoma symptom scale (GSS) questionnaires, breakup time test (BUT), Schirmer I test, corneal and conjunctival staining (Oxford scale), and conjunctival in vivo confocal microscopy (Heidelberg Retina Tomograph 3, Heidelberg Engineering GmbH, Heidelberg, Germany)...
April 23, 2018: Advances in Therapy
Lazar Kopanja, Zorana Kovacevic, Marin Tadic, Monika Cecilija Žužek, Milka Vrecl, Robert Frangež
Detailed shape analysis of cells is important to better understand the physiological mechanisms of toxins and determine their effects on cell morphology. This study aimed to develop a procedure for accurate morphological analysis of cell shape and use it as a tool to estimate toxin activity. With the aim of optimizing the method of cell morphology analysis, we determined the influence of ostreolysin A and pleurotolysin B complex (OlyA/PlyB) on the morphology of murine neuronal NG108-15 cells. A computational method was introduced and successfully applied to quantify morphological attributes of the NG108-15 cell line before and after 30 and 60 min exposure to OlyA/PlyB using confocal microscopy...
April 23, 2018: Histochemistry and Cell Biology
Martin Hruska, Nathan Henderson, Sylvain J Le Marchand, Haani Jafri, Matthew B Dalva
Experience results in long-lasting changes in dendritic spine size, yet how the molecular architecture of the synapse responds to plasticity remains poorly understood. Here a combined approach of multicolor stimulated emission depletion microscopy (STED) and confocal imaging in rat and mouse demonstrates that structural plasticity is linked to the addition of unitary synaptic nanomodules to spines. Spine synapses in vivo and in vitro contain discrete and aligned subdiffraction modules of pre- and postsynaptic proteins whose number scales linearly with spine size...
April 23, 2018: Nature Neuroscience
Shu-Ta Wu, Anthony J Fowler, Corey B Garmon, Adam B Fessler, Joshua D Ogle, Kajal R Grover, Bailey C Allen, Chandra D Williams, Ru Zhou, Mahboubeh Yazdanifar, Craig A Ogle, Pinku Mukherjee
BACKGROUND: Pancreatic ductal adenocarcinoma (PDA) remains the most aggressive cancers with a 5-year survival below 10%. Systemic delivery of chemotherapy drugs has severe side effects in patients with PDA and does not significantly improve overall survival rate. It is highly desirable to advance the therapeutic efficacy of chemotherapeutic drugs by targeting their delivery and increasing accumulation at the tumor site. MUC1 is a membrane-tethered glycoprotein that is aberrantly overexpressed in > 80% of PDA thus making it an attractive antigenic target...
April 23, 2018: BMC Cancer
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