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https://www.readbyqxmd.com/read/28054419/identification-and-initial-characterization-of-matrix-metalloproteinases-in-the-yellow-fever-mosquito-aedes-aegypti
#1
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
#2
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
#3
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
#4
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
#5
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
https://www.readbyqxmd.com/read/27917008/visualization-of-neutrophil-extracellular-traps-and-fibrin-meshwork-in-human-fibrinopurulent-inflammatory-lesions-iii-correlative-light-and-electron-microscopic-study
#6
Takanori Onouchi, Kazuya Shiogama, Yasuyoshi Mizutani, Takashi Takaki, Yutaka Tsutsumi
Neutrophil extracellular traps (NETs) released from dead neutrophils at the site of inflammation represent webs of neutrophilic DNA stretches dotted with granule-derived antimicrobial proteins, including lactoferrin, and play important roles in innate immunity against microbial infection. We have shown the coexistence of NETs and fibrin meshwork in varied fibrinopurulent inflammatory lesions at both light and electron microscopic levels. In the present study, correlative light and electron microscopy (CLEM) employing confocal laser scanning microscopy and scanning electron microscopy was performed to bridge light and electron microscopic images of NETs and fibrin fibrils in formalin-fixed, paraffin-embedded, autopsied lung sections of legionnaire's pneumonia...
November 1, 2016: Acta Histochemica et Cytochemica
https://www.readbyqxmd.com/read/27862974/phenotypic-variation-in-food-utilization-in-an-outbreak-insect-herbivore
#7
Roberto Quezada-García, Álvaro Fuentealba, Éric Bauce
The effects of nutrition may have subtantial impact on insect evolution by shaping different components of phenotypes. The key to undestanding this evolutionary process is to know how nutritional condition affects additive and non-additive components of the phenotype. However, this is poorly understood in outbreaking insects. We investigated the additive and non-additive variation present in food utilization traits in spruce budworm individuals subjected to chronic nutritional stress. 160 full-sib families of spruce budworm (Choristoneura fumiferana Clem...
November 8, 2016: Insect Science
https://www.readbyqxmd.com/read/27855547/traumatic-brain-injury-history-is-associated-with-earlier-age-of-onset-of-alzheimer-disease
#8
Christian LoBue, Hannah Wadsworth, Kristin Wilmoth, Matthew Clem, John Hart, Kyle B Womack, Nyaz Didehbani, Laura H Lacritz, Heidi C Rossetti, C Munro Cullum
OBJECTIVE: This study examined whether a history of traumatic brain injury (TBI) is associated with earlier onset of Alzheimer disease (AD), independent of apolipoprotein ε4 status (Apoe4) and gender. METHOD: Participants with a clinical diagnosis of AD (n = 7625) were obtained from the National Alzheimer's Coordinating Center Uniform Data Set, and categorized based on self-reported lifetime TBI with loss of consciousness (LOC) (TBI+ vs. TBI-) and presence of Apoe4...
January 2017: Clinical Neuropsychologist
https://www.readbyqxmd.com/read/27851087/1451-norepinephrine-and-vasopressin-vs-norepinephrine-alone-for-septic-shock-retrospective-cohort-study
#9
Drayton Hammond, Julia Cullen, Kelsey McCain, Oktawia Clem, Amy Brotherton, Divyan Chopra, Nikhil Meena, Jacob Painter
No abstract text is available yet for this article.
December 2016: Critical Care Medicine
https://www.readbyqxmd.com/read/27851068/1432-discontinuation-of-vasopressin-before-norepinephrine-in-recovery-phase-of-septic-shock-cohort-study
#10
Drayton Hammond, Oktawia Clem, Amy Brotherton, Kelsey McCain, Julia Cullen, Divyan Chopra, Nikhil Meena, Jacob Painter
No abstract text is available yet for this article.
December 2016: Critical Care Medicine
https://www.readbyqxmd.com/read/27850986/1350-norepinephrine-and-vasopressin-vs-norepinephrine-alone-for-septic-shock-randomized-controlled-trial
#11
Oktawia Clem, Jacob Painter, Julia Cullen, Kelsey McCain, Krishna Kakkera, Nikhil Meena, Drayton Hammond
No abstract text is available yet for this article.
December 2016: Critical Care Medicine
https://www.readbyqxmd.com/read/27847357/visualization-of-hiv-t-cell-virological-synapses-and-virus-containing-compartments-by-three-dimensional-correlative-light-and-electron-microscopy
#12
Lili Wang, Edward T Eng, Kenneth Law, Ronald E Gordon, William J Rice, Benjamin K Chen
: Virological synapses (VS) are adhesive structures that form between infected and uninfected cells to enhance the spread of HIV-1. During T cell VS formation, viral proteins are actively recruited to the site of cell-cell contact where the viral material is efficiently translocated to target cells into heterogeneous, protease-resistant, antibody-inaccessible compartments. Using correlative light and electron microscopy (CLEM), we define the membrane topography of the virus-containing compartments (VCC) where HIV is found following VS-mediated transfer...
January 15, 2017: Journal of Virology
https://www.readbyqxmd.com/read/27834401/3-d-em-exploration-of-the-hepatic-microarchitecture-lessons-learned-from-large-volume-in-situ-serial-sectioning
#13
Gerald John Shami, Delfine Cheng, Minh Huynh, Celien Vreuls, Eddie Wisse, Filip Braet
To-date serial block-face scanning electron microscopy (SBF-SEM) dominates as the premier technique for generating three-dimensional (3-D) data of resin-embedded biological samples at an unprecedented depth volume. Given the infancy of the technique, limited literature is currently available regarding the applicability of SBF-SEM for the ultrastructural investigation of tissues. Herein, we provide a comprehensive and rigorous appraisal of five different SBF-SEM sample preparation protocols for the large-volume exploration of the hepatic microarchitecture at an unparalleled X, Y and Z resolution...
November 11, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27832204/expression-of-the-lncrna-maternally-expressed-gene-3-meg3-contributes-to-the-control-of-lung-cancer-cell-proliferation-by-the-rb-pathway
#14
Traci L Kruer, Susan M Dougherty, Lindsey Reynolds, Elizabeth Long, Tanya de Silva, William W Lockwood, Brian F Clem
Maternally expressed gene 3 (MEG3, mouse homolog Gtl2) encodes a long noncoding RNA (lncRNA) that is expressed in many normal tissues, but is suppressed in various cancer cell lines and tumors, suggesting it plays a functional role as a tumor suppressor. Hypermethylation has been shown to contribute to this loss of expression. We now demonstrate that MEG3 expression is regulated by the retinoblastoma protein (Rb) pathway and correlates with a change in cell proliferation. Microarray analysis of mouse embryonic fibroblasts (MEFs) isolated from mice with genetic deletion of all three Rb family members (TKO) revealed a significant silencing of Gtl2/MEG3 expression compared to WT MEFs, and re-expression of Gtl2/MEG3 caused decrease in cell proliferation and increased apoptosis...
2016: PloS One
https://www.readbyqxmd.com/read/27796326/shedding-light-on-cell-compartmentation-in-the-candidate-phylum-poribacteria-by-high-resolution-visualisation-and-transcriptional-profiling
#15
Martin T Jahn, Sebastian M Markert, Taewoo Ryu, Timothy Ravasi, Christian Stigloher, Ute Hentschel, Lucas Moitinho-Silva
Assigning functions to uncultivated environmental microorganisms continues to be a challenging endeavour. Here, we present a new microscopy protocol for fluorescence in situ hybridisation-correlative light and electron microscopy (FISH-CLEM) that enabled, to our knowledge for the first time, the identification of single cells within their complex microenvironment at electron microscopy resolution. Members of the candidate phylum Poribacteria, common and uncultivated symbionts of marine sponges, were used towards this goal...
October 31, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27738022/complete-genome-sequence-of-bacillus-phage-belinda-from-grand-cayman-island
#16
Eileen F Breslin, Jessica Cornell, Zachary Schuhmacher, Madison Himelright, Aya Andos, Ariel Childs, Ana Clem, Monica Gerber, Arissa Gordillo, Laith Harb, Reafa Hossain, Taylor Hutchinson, Isaac Miller, Edmund Morton, Ryan Walters, Destin Webb, Louise Temple
Soil from George Town, Grand Cayman Island, yielded the bacteriophage Belinda, isolated on Bacillus thuringiensis DSM 350. We present here the analysis of the complete genome sequence of 162,308 bp, with 298 predicted genes. The genome also contains three tRNA genes. Belinda belongs to the C1 cluster of Bacillus phages.
October 13, 2016: Genome Announcements
https://www.readbyqxmd.com/read/27613609/inhibition-of-6-phosphofructo-2-kinase-pfkfb3-suppresses-glucose-metabolism-and-the-growth-of-her2-breast-cancer
#17
Julie O'Neal, Amy Clem, Lindsey Reynolds, Susan Dougherty, Yoannis Imbert-Fernandez, Sucheta Telang, Jason Chesney, Brian F Clem
PURPOSE: Human epidermal growth factor receptor-2 (HER2) has been implicated in the progression of multiple tumor types, including breast cancer, and many downstream effectors of HER2 signaling are primary regulators of cellular metabolism, including Ras and Akt. A key downstream metabolic target of Ras and Akt is the 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 isozyme (PFKFB3), whose product, fructose-2,6-bisphosphate (F26BP), is a potent allosteric activator of a rate-limiting enzyme in glycolysis, 6-phosphofructo-1-kinase (PFK-1)...
November 2016: Breast Cancer Research and Treatment
https://www.readbyqxmd.com/read/27609565/ftsz-does-not-initiate-membrane-constriction-at-the-onset-of-division
#18
Daniel O Daley, Ulf Skoglund, Bill Söderström
The source of constriction required for division of a bacterial cell remains enigmatic. FtsZ is widely believed to be a key player, because in vitro experiments indicate that it can deform liposomes when membrane tethered. However in vivo evidence for such a role has remained elusive as it has been challenging to distinguish the contribution of FtsZ from that of peptidoglycan-ingrowth. To differentiate between these two possibilities we studied the early stages of division in Escherichia coli, when FtsZ is present at the division site but peptidoglycan synthesizing enzymes such as FtsI and FtsN are not...
2016: Scientific Reports
https://www.readbyqxmd.com/read/27601992/correlative-light-electron-microscopy-connecting-synaptic-structure-and-function
#19
REVIEW
Isabell Begemann, Milos Galic
Many core paradigms of contemporary neuroscience are based on information obtained by electron or light microscopy. Intriguingly, these two imaging techniques are often viewed as complementary, yet separate entities. Recent technological advancements in microscopy techniques, labeling tools, and fixation or preparation procedures have fueled the development of a series of hybrid approaches that allow correlating functional fluorescence microscopy data and ultrastructural information from electron micrographs from a singular biological event...
2016: Frontiers in Synaptic Neuroscience
https://www.readbyqxmd.com/read/27576708/correlative-light-and-electron-microscopy-of-nucleolar-transcription-in-saccharomyces-cerevisiae
#20
Christophe Normand, Maxime Berthaud, Olivier Gadal, Isabelle Léger-Silvestre
Nucleoli form around RNA polymerase I transcribed ribosomal RNA (rRNA) genes. The direct electron microscopy observation of rRNA genes after nucleolar chromatin spreading (Miller's spreads) constitutes to date the only system to quantitatively assess transcription at a single molecule level. However, the spreading procedure is likely generating artifact and despite being informative, these spread rRNA genes are far from their in vivo situation. The integration of the structural characterization of spread rRNA genes in the three-dimensional (3D) organization of the nucleolus would represent an important scientific achievement...
2016: Methods in Molecular Biology
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