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https://www.readbyqxmd.com/read/28435429/antibacterial-and-cytotoxic-activities-of-naphthoquinone-pigments-from-onosma-visianii-clem
#1
Milena D Vukic, Nenad L Vukovic, Gorica T Djelic, Suzana Lj Popovic, Milan M Zaric, Dejan D Baskic, Gordana B Krstic, Vele V Tesevic, Miroslava M Kacaniova
In this study, the antibacterial and cytotoxic activities of isolated compounds from the roots of Onosma visianii were investigated. By using different chromatographic techniques and appropriate spectroscopic methods, the seven naphthoquinones were described: deoxyshikonin ( 1 ), isobutyrylshikonin ( 2 ), α-methylbutyrylshikonin ( 3 ), acetylshikonin ( 4 ), β-hydroxyisovalerylshikonin ( 5 ), 5,8-O-dimethyl isobutyrylshikonin ( 6 ) and 5,8-O-dimethyl deoxyshikonin ( 7 ). Among the tested compounds, 3 and 4 exhibited the highest antibacterial activities toward all tested bacterial species (MIC50 and MIC90 for gram positive bacteria: 6...
2017: EXCLI journal
https://www.readbyqxmd.com/read/28430270/atomistic-non-adiabatic-dynamics-of-the-lh2-complex-with-a-gpu-accelerated-ab-initio-exciton-model
#2
Aaron Sisto, Clem Stross, Marc W van der Kamp, Michael O'Connor, Simon McIntosh-Smith, Graham T Johnson, Edward G Hohenstein, Fred R Manby, David R Glowacki, Todd J Martinez
We recently outlined an efficient multi-tiered parallel ab initio excitonic framework that utilizes time dependent density functional theory (TDDFT) to calculate ground and excited state energies and gradients of large supramolecular complexes in atomistic detail - enabling us to undertake non-adiabatic simulations which explicitly account for the coupled anharmonic vibrational motion of all the constituent atoms in a supramolecular system. Here we apply that framework to the 27 coupled bacterio-chlorophyll-a chromophores which make up the LH2 complex, using it to compute an on-the-fly nonadiabatic surface-hopping (SH) trajectory of electronically excited LH2...
April 21, 2017: Physical Chemistry Chemical Physics: PCCP
https://www.readbyqxmd.com/read/28429122/quantifying-golgi-structure-using-em-combining-volume-sem-and-stereology-for-higher-throughput
#3
REVIEW
Sophie Ferguson, Anna M Steyer, Terry M Mayhew, Yannick Schwab, John Milton Lucocq
Investigating organelles such as the Golgi complex depends increasingly on high-throughput quantitative morphological analyses from multiple experimental or genetic conditions. Light microscopy (LM) has been an effective tool for screening but fails to reveal fine details of Golgi structures such as vesicles, tubules and cisternae. Electron microscopy (EM) has sufficient resolution but traditional transmission EM (TEM) methods are slow and inefficient. Newer volume scanning EM (volume-SEM) methods now have the potential to speed up 3D analysis by automated sectioning and imaging...
April 20, 2017: Histochemistry and Cell Biology
https://www.readbyqxmd.com/read/28425495/the-biglen-gpr171-peptide-receptor-system-within-the-basolateral-amygdala-regulates-anxiety-like-behavior-and-contextual-fear-conditioning
#4
Erin N Bobeck, Ivone Gomes, Darlene Pena, Kirstie A Cummings, Roger L Clem, Mihaly Mezei, Lakshmi A Devi
Studies show that neuropeptide-receptor systems in the basolateral amygdala (BLA) play an important role in the pathology of anxiety and other mood disorders. Since GPR171, a recently deorphanized receptor for the abundant neuropeptide BigLEN, is expressed in the BLA, we investigated its role in fear and anxiety-like behaviors. To carry out these studies we identified small molecule ligands using a homology model of GPR171 to virtually screen a library of compounds. One of the hits, MS0021570_1, was identified as a GPR171 antagonist based on its ability to block (i) BigLEN-mediated activation of GPR171 in heterologous cells, (ii) BigLEN-mediated hyperpolarization of BLA pyramidal neurons, and (iii) feeding induced by DREADD-mediated activation of BigLEN containing AgRP neurons in the arcuate nucleus...
April 20, 2017: Neuropsychopharmacology: Official Publication of the American College of Neuropsychopharmacology
https://www.readbyqxmd.com/read/28384138/diverse-protocols-for-correlative-super-resolution-fluorescence-imaging-and-electron-microscopy-of-chemically-fixed-samples
#5
Benjamin G Kopek, Maria G Paez-Segala, Gleb Shtengel, Kem A Sochacki, Mei G Sun, Yalin Wang, C Shan Xu, Schuyler B van Engelenburg, Justin W Taraska, Loren L Looger, Harald F Hess
Our groups have recently developed related approaches for sample preparation for super-resolution imaging within endogenous cellular environments using correlative light and electron microscopy (CLEM). Four distinct techniques for preparing and acquiring super-resolution CLEM data sets for aldehyde-fixed specimens are provided, including Tokuyasu cryosectioning, whole-cell mount, cell unroofing and platinum replication, and resin embedding and sectioning. The choice of the best protocol for a given application depends on a number of criteria that are discussed in detail...
May 2017: Nature Protocols
https://www.readbyqxmd.com/read/28361485/the-use-of-correlative-light-electron-microscopy-clem-to-study-pink1-parkin-mediated-mitophagy
#6
Chieko Kishi-Itakura, Folma Buss
In this chapter we describe the use of correlative light-electron microscopy (CLEM) to study, in cultured cells, the turnover of damaged mitochondria by PINK1/Parkin-dependent mitophagy. CLEM combines the advantages of light microscopy, which allows to image and rapidly screen a large number of the cells, while electron microscopy provides high-resolution imaging of these selected cells and a detailed structural analysis of their cellular organelles. We describe in detail how to prepare the cell cultures for optimum preservation of their cellular ultrastructure for CLEM using the most suitable buffers, fixatives, and embedding resins...
March 31, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28348028/examination-of-e-faecalis-toxin-antitoxin-ta-toxin-fst-function-utilizing-a-pheromone-inducible-expression-vector-with-tight-repression-and-large-dynamic-range
#7
Keith E Weaver, Yuqing Chen, Elly M Miiller, Jake N Johnson, Alex A Dangler, Dawn A Manias, Aaron M Clem, Daniel J Schjodt, Gary M Dunny
Tools for regulated gene expression in E. faecalis are extremely limited. In this study we describe the construction of an expression vector for E. faecalis, designated pCIE, utilizing the PQ pheromone-responsive promoter of plasmid pCF10. We demonstrate that this promoter is tightly repressed, responds to nanogram quantities of the peptide pheromone, and has a large dynamic range. To demonstrate its utility, the promoter was used to control expression of the toxic peptides of two par family toxin-antitoxin loci present in E...
March 27, 2017: Journal of Bacteriology
https://www.readbyqxmd.com/read/28347808/optimizing-clem-protocols-for-plants-cells-gma-embedding-and-cryosections-as-alternatives-for-preservation-of-gfp-fluorescence-in-arabidopsis-roots
#8
Jessica Marion, Romain Le Bars, Béatrice Satiat-Jeunemaitre, Claire Boulogne
Recently, a number of diverse correlative light and electron microscopy (CLEM) protocols have been developed for several model organisms. However, these CLEM methods have largely bypassed plant cell research, with most protocols having little application to plants. Using autophagosome identification as a biological background, we propose and compare two CLEM protocols that can be performed in most plant research laboratories, providing a good compromise that preserves fluorescent signals as well as ultrastructural features...
March 24, 2017: Journal of Structural Biology
https://www.readbyqxmd.com/read/28323681/predictors-that-a-diagnosis-of-mild-cognitive-impairment-will-remain-stable-3-years-later
#9
Matthew A Clem, Ryan P Holliday, Seema Pandya, Linda S Hynan, Laura H Lacritz, Fu L Woon
BACKGROUND AND OBJECTIVE: In half to two thirds of patients who are diagnosed with mild cognitive impairment (MCI), the diagnosis neither converts to dementia nor reverts to normal cognition; however, little is known about predictors of MCI stability. Our study aimed to identify those predictors. METHODS: We obtained 3-year longitudinal data from the National Alzheimer's Coordinating Center Uniform Data Set for patients with a baseline diagnosis of MCI. To predict MCI stability, we used the patients' baseline data to conduct three logistic regression models: demographics, global function, and neuropsychological performance...
March 2017: Cognitive and Behavioral Neurology: Official Journal of the Society for Behavioral and Cognitive Neurology
https://www.readbyqxmd.com/read/28317888/direct-evidence-of-lack-of-colocalisation-of-fluorescently-labelled-gold-labels-used-in-correlative-light-electron-microscopy
#10
Benjamin T Miles, Alexander B Greenwood, David Benito-Alifonso, Hugh Tanner, M Carmen Galan, Paul Verkade, Henkjan Gersen
Fluorescently labelled nanoparticles are routinely used in Correlative Light Electron Microscopy (CLEM) to combine the capabilities of two separate microscope platforms: fluorescent light microscopy (LM) and electron microscopy (EM). The inherent assumption is that the fluorescent label observed under LM colocalises well with the electron dense nanoparticle observed in EM. Herein we show, by combining single molecule fluorescent imaging with optical detection of the scattering from single gold nanoparticles, that for a commercially produced sample of 10 nm gold nanoparticles tagged to Alexa-633 there is in fact no colocalisation between the fluorescent signatures of Alexa-633 and the scattering associated with the gold nanoparticle...
March 20, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28278087/laccase-immobilized-dendritic-nanofibrous-membranes-as-a-novel-approach-towards-the-removal-of-bisphenol-a
#11
Lebohang E Koloti, Nonjabulo P Gule, Omotayo A Arotiba, Soraya P Malinga
Laccase enzymes from Rhus vernificera were covalently bound on hyperbranched polyethyleneimine/polyethersulfone (HPEI/PES) electrospun nanofibrous membranes and used for the removal of bisphenol A (BPA) from water. The laccase enzyme was anchored on the dendritic membranes through the abundant peripheral amine groups on the hyperbranched polyethyleneimine (HPEI) using glutaraldehyde as a cross-linker. The membranes were characterised with attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy, scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy (SEM-EDS) and ultraviolet-visible (UV-Vis) spectroscopy and correlative light and electron microscopy (CLEM)...
March 2, 2017: Environmental Technology
https://www.readbyqxmd.com/read/28230492/clem-analysis-of-the-angiogenic-process-in-a-zebrafish-model-of-spinal-cord-injury
#12
A Pinto, A Ribeiro, T Carvalho, M L Saúde
No abstract text is available yet for this article.
January 2017: Ultrastructural Pathology
https://www.readbyqxmd.com/read/28193691/optogenetic-examination-of-prefrontal-amygdala-synaptic-development
#13
Maithe Arruda-Carvalho, Wan-Chen Wu, Kirstie A Cummings, Roger L Clem
A brain network comprising the medial prefrontal cortex (mPFC) and amygdala plays important roles in developmentally regulated cognitive and emotional processes. However, very little is known about the maturation of mPFC-amygdala circuitry. We conducted anatomical tracing of mPFC projections and optogenetic interrogation of their synaptic connections with neurons in the basolateral amygdala (BLA) at neonatal to adult developmental stages in mice. Results indicate that mPFC-BLA projections exhibit delayed emergence relative to other mPFC pathways and establish synaptic transmission with BLA excitatory and inhibitory neurons in late infancy, events that coincide with a massive increase in overall synaptic drive...
March 15, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/28154417/a-global-approach-for-quantitative-super-resolution-and-electron-microscopy-on-cryo-and-epoxy-sections-using-self-labeling-protein-tags
#14
Andreas Müller, Martin Neukam, Anna Ivanova, Anke Sönmez, Carla Münster, Susanne Kretschmar, Yannis Kalaidzidis, Thomas Kurth, Jean-Marc Verbavatz, Michele Solimena
Correlative light and electron microscopy (CLEM) is a powerful approach to investigate the molecular ultrastructure of labeled cell compartments. However, quantitative CLEM studies are rare, mainly due to small sample sizes and the sensitivity of fluorescent proteins to strong fixatives and contrasting reagents for EM. Here, we show that fusion of a self-labeling protein to insulin allows for the quantification of age-distinct insulin granule pools in pancreatic beta cells by a combination of super resolution and transmission electron microscopy on Tokuyasu cryosections...
December 2017: Scientific Reports
https://www.readbyqxmd.com/read/28139674/ec-clem-flexible-multidimensional-registration-software-for-correlative-microscopies
#15
Perrine Paul-Gilloteaux, Xavier Heiligenstein, Martin Belle, Marie-Charlotte Domart, Banafshe Larijani, Lucy Collinson, Graça Raposo, Jean Salamero
No abstract text is available yet for this article.
January 31, 2017: Nature Methods
https://www.readbyqxmd.com/read/28054419/identification-and-initial-characterization-of-matrix-metalloproteinases-in-the-yellow-fever-mosquito-aedes-aegypti
#16
A M Kantor, S Dong, N L Held, E Ishimwe, A L Passarelli, R J Clem, A W E Franz
Aedes aegypti is a major vector for arboviruses such as dengue, chikungunya and Zika viruses. During acquisition of a viremic bloodmeal, an arbovirus infects mosquito midgut cells before disseminating to secondary tissues, including the salivary glands. Once virus is released into the salivary ducts it can be transmitted to another vertebrate host. The midgut is surrounded by a basal lamina (BL) in the extracellular matrix, consisting of a proteinaceous mesh composed of collagen IV and laminin. BL pore size exclusion limit prevents virions from passing through...
February 2017: Insect Molecular Biology
https://www.readbyqxmd.com/read/28011168/toxoplasma-gondii-plaque-assays-revisited-improvements-for-ultrastructural-and-quantitative-evaluation-of-lytic-parasite-growth
#17
Christoph-Martin Ufermann, Florian Müller, Nora Frohnecke, Michael Laue, Frank Seeber
Lytic growth of intracellular Toxoplasma gondii tachyzoite stages over a period of days results in plaques within mononolayers of host cells. Plaque assays are in frequent use to isolate single clones and to investigate invasion, replication and egress over a longer time frame. To allow correlating plaque morphology and/or size with ultrastructural examination of individual parasites we introduce a simple protocol for correlative light and electron microscopy (CLEM) of entire plaques. We also illustrate the advantages of visualizing only the boundaries of plaques by staining for infected cells ('positive staining') rather than the traditional staining of the intact cell monolayer, thus outlining the area of lysed cells ('negative staining')...
December 21, 2016: Experimental Parasitology
https://www.readbyqxmd.com/read/27977021/correlated-fluorescence-microscopy-and-cryo-electron-tomography-of-virus-infected-or-transfected-mammalian-cells
#18
Cheri M Hampton, Joshua D Strauss, Zunlong Ke, Rebecca S Dillard, Jason E Hammonds, Eric Alonas, Tanay M Desai, Mariana Marin, Rachel E Storms, Fredrick Leon, Gregory B Melikyan, Philip J Santangelo, Paul W Spearman, Elizabeth R Wright
Correlative light and electron microscopy (CLEM) combines spatiotemporal information from fluorescence light microscopy (fLM) with high-resolution structural data from cryo-electron tomography (cryo-ET). These technologies provide opportunities to bridge knowledge gaps between cell and structural biology. Here we describe our protocol for correlated cryo-fLM, cryo-electron microscopy (cryo-EM), and cryo-ET (i.e., cryo-CLEM) of virus-infected or transfected mammalian cells. Mammalian-derived cells are cultured on EM substrates, using optimized conditions that ensure that the cells are spread thinly across the substrate and are not physically disrupted...
January 2017: Nature Protocols
https://www.readbyqxmd.com/read/27974202/the-drosophila-clock-neuron-network-features-diverse-coupling-modes-and-requires-network-wide-coherence-for-robust-circadian-rhythms
#19
Zepeng Yao, Amelia J Bennett, Jenna L Clem, Orie T Shafer
In animals, networks of clock neurons containing molecular clocks orchestrate daily rhythms in physiology and behavior. However, how various types of clock neurons communicate and coordinate with one another to produce coherent circadian rhythms is not well understood. Here, we investigate clock neuron coupling in the brain of Drosophila and demonstrate that the fly's various groups of clock neurons display unique and complex coupling relationships to core pacemaker neurons. Furthermore, we find that coordinated free-running rhythms require molecular clock synchrony not only within the well-characterized lateral clock neuron classes but also between lateral clock neurons and dorsal clock neurons...
December 13, 2016: Cell Reports
https://www.readbyqxmd.com/read/27943680/progression-of-lung-disease-in-preschool-patients-with-cystic-fibrosis
#20
Sanja Stanojevic, Stephanie D Davis, George Retsch-Bogart, Hailey Webster, Miriam Davis, Robin C Johnson, Renee Jensen, Maria Ester Pizarro, Mica Kane, Charles C Clem, Leah Schornick, Padmaja Subbarao, Felix A Ratjen
RATIONALE: Implementation of intervention strategies to prevent lung damage in early cystic fibrosis (CF) requires objective outcome measures that capture and track lung disease. OBJECTIVE: To define the utility of the Lung Clearance Index (LCI), measured by Multiple Breath Washout (MBW), as a means to track disease progression in preschool children with CF. METHODS: CF children between the ages of 2.5 and 6 years with a confirmed diagnosis of CF and age-matched healthy controls were enrolled at three North American CF centers...
December 12, 2016: American Journal of Respiratory and Critical Care Medicine
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