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https://www.readbyqxmd.com/read/28811466/time-lapse-crystallography-snapshots-of-a-double-strand-break-repair-polymerase-in-action
#1
Joonas A Jamsen, William A Beard, Lars C Pedersen, David D Shock, Andrea F Moon, Juno M Krahn, Katarzyna Bebenek, Thomas A Kunkel, Samuel H Wilson
DNA polymerase (pol) μ is a DNA-dependent polymerase that incorporates nucleotides during gap-filling synthesis in the non-homologous end-joining pathway of double-strand break repair. Here we report time-lapse X-ray crystallography snapshots of catalytic events during gap-filling DNA synthesis by pol μ. Unique catalytic intermediates and active site conformational changes that underlie catalysis are uncovered, and a transient third (product) metal ion is observed in the product state. The product manganese coordinates phosphate oxygens of the inserted nucleotide and PPi...
August 15, 2017: Nature Communications
https://www.readbyqxmd.com/read/28808850/non-rem-sleep-disordered-breathing-affects-performance-on-the-psychomotor-vigilance-task
#2
Takuro Kitamura, Soichiro Miyazaki, Hiroshi Kadotani, Takashi Kanemura, Harun Bin Sulaiman, Shoko Takeuchi, Takahisa Tabata, Hideaki Suzuki
PURPOSE: Although many studies have investigated the clinical importance of sleep apnea on rapid eye movement (REM) and non-REM (NREM) sleep, the relationship between behavioral performance and apneic events during different sleep phases remains unclear. In the present study, we sought to investigate the effect of sleep phase fragmentation due to sleep-disordered breathing (SDB) during REM and NREM on the vigilance and sustainability of attention based on psychomotor vigilance task (PVT) performance...
August 14, 2017: Sleep & Breathing, Schlaf & Atmung
https://www.readbyqxmd.com/read/28808446/neuron-specific-fluorescence-reporter-based-live-cell-tracing-for-transdifferentiation-of-mesenchymal-stem-cells-into-neurons-by-chemical-compound
#3
Do Won Hwang, Hyun Woo Kwon, Jaeho Jang, Hee Jung Jung, Kwang Rok Kim, Dong Soo Lee
Although transdifferentiation of mesenchymal stem cells (MSCs) into neurons increases the possibility of therapeutic use of MSCs for neurodevelopmental disorders, the use of MSCs has the limitation on differentiation efficiency to neuronal lineage and lack of an easy method to monitor the transdifferentiation. In this study, using time-lapse live cell imaging, we assessed the neuronal differentiation of MSCs induced by a small molecule "NHPDQC (N-hydroxy-2-oxo-3-(3-phenylprophyl)-1,2-dihydroquinoxaline-6-carboxamide, C18H17N3O3)...
2017: Stem Cells International
https://www.readbyqxmd.com/read/28807036/t-time-a-data-repository-of-t-cell-and-calcium-release-activated-calcium-channel-activation-imagery
#4
Cody Arbuckle, Milton Greenberg, Adrienne Bergh, Rene German, Nick Sirago, Erik Linstead
BACKGROUND: A fundamental understanding of live-cell dynamics is necessary in order to advance scientific techniques and personalized medicine. For this understanding to be possible, image processing techniques, probes, tracking algorithms and many other methodologies must be improved. Currently there are no large open-source datasets containing live-cell imaging to act as a standard for the community. As a result, researchers cannot evaluate their methodologies on an independent benchmark or leverage such a dataset to formulate scientific questions...
August 15, 2017: BMC Research Notes
https://www.readbyqxmd.com/read/28803829/the-vascular-niche-regulates-hematopoietic-stem-and-progenitor-cell-lodgment-and-expansion-via-klf6a-ccl25b
#5
Yuanyuan Xue, Junhua Lv, Chunxia Zhang, Lu Wang, Dongyuan Ma, Feng Liu
In mammals, hematopoietic stem and progenitor cells (HSPCs) rapidly expand in the fetal liver (FL), but the underlying mechanism remains unclear. Here, we characterize zebrafish caudal hematopoietic tissue (CHT) and identify an important cellular and molecular mechanism of HSPC expansion. Time-lapse imaging showed that HSPCs localize adjacent to vascular endothelial cells (ECs), and their migration and expansion display caudal vein-specific orientation in the CHT. RNA sequencing and functional analysis identified that an EC-expressed transcription factor, Krüppel-like factor 6a (Klf6a), is essential for the CHT niche...
August 9, 2017: Developmental Cell
https://www.readbyqxmd.com/read/28802829/embryonic-zebrafish-primary-cell-culture-for-transfection-and-live-cellular-and-subcellular-imaging
#6
Wiebke A Sassen, Franziska Lehne, Giulio Russo, Sven Wargenau, Stefan Dübel, Reinhard W Köster
Although having great potential for live cell imaging to address numerous cell biological questions with high spatial and temporal resolution, primary cell cultures of zebrafish embryos are not widely used. We present an easy-to-use protocol for preparing primary cell cultures of 2 dpf zebrafish embryos allowing for live cell imaging of fully differentiated cells such as neurons and myocytes. We demonstrate that different cell types can be identified by morphology and expression of transgenic cell type-specific fluorescent reporters and that fluorescent cells can be sorted by flow cytometry to prepare an enriched culture...
August 9, 2017: Developmental Biology
https://www.readbyqxmd.com/read/28802391/pathophysiology-of-esophageal-impairment-due-to-button-battery-ingestion
#7
Johannes Völker, Christine Völker, Philipp Schendzielorz, Sebastian P Schraven, Andreas Radeloff, Robert Mlynski, Rudolf Hagen, Kristen Rak
BACKGROUND: The increased use of button batteries with high energy densities in devices of daily life presents a high risk of injury, especially for toddlers and young children. If an accidental ingestion of a button battery occurs, this foreign body can become caught in the constrictions of the esophagus and cause serious damage to the adjacent tissue layers. The consequences can be ulcerations, perforations with fistula formation and damage to the surrounding anatomical structures. In order to gain a better understanding of the pathophysiology after ingestion, we carried out systematic studies on fresh preparations of porcine esophagi...
September 2017: International Journal of Pediatric Otorhinolaryngology
https://www.readbyqxmd.com/read/28799616/quantitative-investigation-of-mda-mb-231-breast-cancer-cell-motility-dependence-on-epidermal-growth-factor-concentration-and-its-gradient
#8
Tanzila Islam, Haluk Resat
Enhanced cell motility is one of the primary features of cancer. Accumulated evidence demonstrates that Epidermal Growth Factor Receptor (EGFR) mediated pathways play an important role in breast cancer cell proliferation and migration. We have quantified the MDA-MB-231 breast cancer cell migration in response to the stimulation of EGFR pathways with their ligand EGF to determine how the cell motility of MDA-MB-231 cells depends on the ligand concentration and gradient. Analysis at the single cell level combined with mathematical modeling and the ability to vary the ligand concentration and gradients locally using microfluidic devices allowed us to separate the unique contributions of ligand concentration and ligand gradient to cell motility...
August 11, 2017: Molecular BioSystems
https://www.readbyqxmd.com/read/28798144/frontline-science-leishmania-mexicana-amastigotes-can-replicate-within-neutrophils
#9
Benjamin P Hurrell, Manon Beaumann, Sandrina Heyde, Ivo B Regli, Andreas J Müller, Fabienne Tacchini-Cottier
Cutaneous leishmaniasis is a neglected tropical disease, causing a spectrum of clinical manifestations varying from self-healing to unhealing lesions that may be very difficult to treat. Emerging evidence points to a detrimental role for neutrophils during the first hours following infection with many distinct Leishmania species (spp.) at a time when the parasite is in its nonreplicative promastigote form. Neutrophils have also been detected at later stages of infection in unhealing chronic cutaneous lesions...
August 10, 2017: Journal of Leukocyte Biology
https://www.readbyqxmd.com/read/28798065/self-organization-process-in-newborn-skin-organoid-formation-inspires-strategy-to-restore-hair-regeneration-of-adult-cells
#10
Mingxing Lei, Linus J Schumacher, Yung-Chih Lai, Wen-Tau Juan, Chao-Yuan Yeh, Ping Wu, Ting-Xin Jiang, Ruth E Baker, Randall Bruce Widelitz, Li Yang, Cheng-Ming Chuong
Organoids made from dissociated progenitor cells undergo tissue-like organization. This in vitro self-organization process is not identical to embryonic organ formation, but it achieves a similar phenotype in vivo. This implies genetic codes do not specify morphology directly; instead, complex tissue architectures may be achieved through several intermediate layers of cross talk between genetic information and biophysical processes. Here we use newborn and adult skin organoids for analyses. Dissociated cells from newborn mouse skin form hair primordia-bearing organoids that grow hairs robustly in vivo after transplantation to nude mice...
August 10, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28797528/actinin-1-and-actinin-4-play-essential-but-distinct-roles-in-invadopodia-formation-by-carcinoma-cells
#11
Hideki Yamaguchi, Yuumi Ito, Nami Miura, Yuko Nagamura, Ayaka Nakabo, Kiyoko Fukami, Kazufumi Honda, Ryuichi Sakai
Invadopodia are ventral membrane protrusions formed by cancer cells that degrade the extracellular matrix (ECM) during tumor invasion and metastasis. Formation of invadopodia is initiated by the assembly of actin filaments (F-actin) that results from the coordinated activation of several actin regulatory proteins. Actinin-1 and actinin-4 are actin bundling proteins expressed in non-muscle cells and actinin-4 is preferentially associated with malignant phenotypes of carcinoma cells. In this study, we investigated the role of actinin-1 and -4 in invadopodia formation...
August 1, 2017: European Journal of Cell Biology
https://www.readbyqxmd.com/read/28795134/in-vivo-analysis-of-the-neurovascular-niche-in-the-developing-xenopus-brain
#12
Melissa Lau, Jianli Li, Hollis T Cline
The neurovascular niche is a specialized microenvironment formed by the interactions between neural progenitor cells (NPCs) and the vasculature. While it is thought to regulate adult neurogenesis by signaling through vascular-derived soluble cues or contacted-mediated cues, less is known about the neurovascular niche during development. In Xenopus laevis tadpole brain, NPCs line the ventricle and extend radial processes tipped with endfeet to the vascularized pial surface. Using in vivo labeling and time-lapse imaging in tadpoles, we find that intracardial injection of fluorescent tracers rapidly labels Sox2/3-expressing NPCs and that vascular-circulating molecules are endocytosed by NPC endfeet...
July 2017: ENeuro
https://www.readbyqxmd.com/read/28795048/evaluation-of-human-embryo-development-in-in-vitro-fertilization-and-intracytoplasmic-sperm-injection-fertilized-oocytes-a-time-lapse-study
#13
Hyung Jun Kim, Hye Jin Yoon, Jung Mi Jang, Won Don Lee, San Hyun Yoon, Jin Ho Lim
OBJECTIVE: We investigated whether the insemination method (in vitro fertilization [IVF] or intracytoplasmic sperm injection [ICSI]) affected morphokinetic events and abnormal cleavage events in embryonic development. METHODS: A total of 1,830 normal fertilized embryos were obtained from 272 IVF and ICSI cycles that underwent ovum retrieval culture using a time-lapse system (Embryoscope) from June 2013 to March 2015. All embryos were investigated by a detailed time-lapse analysis that measured the developmental events in the hours after IVF or ICSI insemination...
June 2017: Clinical and Experimental Reproductive Medicine
https://www.readbyqxmd.com/read/28794472/high-resolution-transport-of-intensity-quantitative-phase-microscopy-with-annular-illumination
#14
Chao Zuo, Jiasong Sun, Jiaji Li, Jialin Zhang, Anand Asundi, Qian Chen
For quantitative phase imaging (QPI) based on transport-of-intensity equation (TIE), partially coherent illumination provides speckle-free imaging, compatibility with brightfield microscopy, and transverse resolution beyond coherent diffraction limit. Unfortunately, in a conventional microscope with circular illumination aperture, partial coherence tends to diminish the phase contrast, exacerbating the inherent noise-to-resolution tradeoff in TIE imaging, resulting in strong low-frequency artifacts and compromised imaging resolution...
August 9, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28792434/collective-migration-of-lens-epithelial-cell-induced-by-differential-microscale-groove-patterns
#15
Chunga Kwon, Youngjun Kim, Hojeong Jeon
Herein, a micro-patterned cell adhesive surface is prepared for the future design of medical devices. One-dimensional polydimethylsiloxane (PDMS) micro patterns were prepared by a photolithography process. We investigated the effect of microscale topographical patterned surfaces on decreasing the collective cell migration rate. PDMS substrates were prepared through soft lithography using Si molds fabricated by photolithography. Afterwards, we observed the collective cell migration of human lens epithelial cells (B-3) on various groove/ridge patterns and evaluated the migration rate to determine the pattern most effective in slowing down the cell sheet spreading speed...
August 9, 2017: Journal of Functional Biomaterials
https://www.readbyqxmd.com/read/28790449/single-cell-analyses-identify-bioengineered-niches-for-enhanced-maintenance-of-hematopoietic-stem-cells
#16
Aline Roch, Sonja Giger, Mukul Girotra, Vasco Campos, Nicola Vannini, Olaia Naveiras, Samy Gobaa, Matthias P Lutolf
The in vitro expansion of long-term hematopoietic stem cells (HSCs) remains a substantial challenge, largely because of our limited understanding of the mechanisms that control HSC fate choices. Using single-cell multigene expression analysis and time-lapse microscopy, here we define gene expression signatures and cell cycle hallmarks of murine HSCs and the earliest multipotent progenitors (MPPs), and analyze systematically single HSC fate choices in culture. Our analysis revealed twelve differentially expressed genes marking the quiescent HSC state, including four genes encoding cell-cell interaction signals in the niche...
August 9, 2017: Nature Communications
https://www.readbyqxmd.com/read/28790202/tethered-magnets-are-the-key-to-magnetotaxis-direct-observations-of-magnetospirillum-magneticum-amb-1-show-that-mamk-distributes-magnetosome-organelles-equally-to-daughter-cells
#17
Azuma Taoka, Ayako Kiyokawa, Chika Uesugi, Yousuke Kikuchi, Zachery Oestreicher, Kaori Morii, Yukako Eguchi, Yoshihiro Fukumori
Magnetotactic bacteria are a unique group of bacteria that synthesize a magnetic organelle termed the magnetosome, which they use to assist with their magnetic navigation in a specific type of bacterial motility called magneto-aerotaxis. Cytoskeletal filaments consisting of the actin-like protein MamK are associated with the magnetosome chain. Previously, the function of MamK was thought to be in positioning magnetosome organelles; this was proposed based on observations via electron microscopy still images...
August 8, 2017: MBio
https://www.readbyqxmd.com/read/28783625/non-rigid-contour-based-registration-of-cell-nuclei-in-2d-live-cell-microscopy-images-using-a-dynamic-elasticity-model
#18
Dmitry V Sorokin, Igor Peterlik, Marco Tektonidis, Karl Rohr, Pavel Matula
The analysis of the pure motion of subnuclear structures without influence of the cell nucleus motion and deformation is essential in live cell imaging. In this work, we propose a 2D contour-based image registration approach for compensation of nucleus motion and deformation in fluorescence microscopy time-lapse sequences. The proposed approach extends our previous approach which uses a static elasticity model to register cell images. Compared to that scheme, the new approach employs a dynamic elasticity model for forward simulation of nucleus motion and deformation based on the motion of its contours...
July 31, 2017: IEEE Transactions on Medical Imaging
https://www.readbyqxmd.com/read/28782074/a-cell-culture-technique-for-human-epiretinal-membranes-to-describe-cell-behavior-and-membrane-contraction-in-vitro
#19
Christian Wertheimer, Kirsten H Eibl-Lindner, Denise Compera, Alexander Kueres, Armin Wolf, Denitsa Docheva, Siegfried G Priglinger, Claudia Priglinger, Ricarda G Schumann
PURPOSE: To introduce a human cell culture technique for investigating in-vitro behavior of primary epiretinal cells and membrane contraction of fibrocellular tissue surgically removed from eyes with idiopathic macular pucker. METHODS: Human epiretinal membranes were harvested from ten eyes with idiopathic macular pucker during standard vitrectomy. Specimens were fixed on cell culture plastic using small entomological pins to apply horizontal stress to the tissue, and then transferred to standard cell culture conditions...
August 7, 2017: Graefe's Archive for Clinical and Experimental Ophthalmology
https://www.readbyqxmd.com/read/28777446/characterization-of-a-novel-eb1-acetylation-site-important-for-the-regulation-of-microtubule-dynamics-and-cargo-recruitment
#20
Songbo Xie, Yang Yang, Xiaochen Lin, Jun Zhou, Dengwen Li, Min Liu
Microtubule plus ends undergo highly dynamic modifications to regulate different aspects of cellular activities. Most microtubule plus-end tracking proteins (+TIPs) are recruited to the microtubule ends by the master loading factor, end-binding protein 1 (EB1). These proteins coordinately regulate microtubule dynamics and cellular plasticity. Acetylation is known to modulate EB1 function; however, the molecular details of EB1 acetylation remain largely unclear. We mapped the acetylation pattern of EB1 and identified several previously uncharacterized sites of EB1 acetylation...
August 4, 2017: Journal of Cellular Physiology
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