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https://www.readbyqxmd.com/read/28108654/revealing-the-role-of-the-product-metal-in-dna-polymerase-%C3%AE-catalysis
#1
Lalith Perera, Bret D Freudenthal, William A Beard, Lee G Pedersen, Samuel H Wilson
DNA polymerases catalyze a metal-dependent nucleotidyl transferase reaction during extension of a DNA strand using the complementary strand as a template. The reaction has long been considered to require two magnesium ions. Recently, a third active site magnesium ion was identified in some DNA polymerase product crystallographic structures, but its role is not known. Using quantum mechanical/ molecular mechanical calculations of polymerase β, we find that a third magnesium ion positioned near the newly identified product metal site does not alter the activation barrier for the chemical reaction indicating that it does not have a role in the forward reaction...
January 20, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28108477/quantitative-microscopy-uncovers-ploidy-changes-during-mitosis-in-live-drosophila-embryos-and-their-effect-on-nuclear-size
#2
Wee Choo Puah, Rambabu Chinta, Martin Wasser
Time-lapse microscopy is a powerful tool to investigate cellular and developmental dynamics. In Drosophila melanogaster, it can be used to study division cycles in embryogenesis. To obtain quantitative information from 3D time-lapse data and track proliferating nuclei from the syncytial stage until gastrulation, we developed an image analysis pipeline consisting of nuclear segmentation, tracking, annotation and quantification. Image analysis of maternal-haploid (mh) embryos revealed that a fraction of haploid syncytial nuclei fused to give rise to nuclei of higher ploidy (2n, 3n, 4n)...
January 20, 2017: Biology Open
https://www.readbyqxmd.com/read/28108476/dynamic-analysis-of-the-mesenchymal-epithelial-transition-of-blood-brain-barrier-forming-glia-in-drosophila
#3
Tina Schwabe, Xiaoling Li, Ulrike Gaul
During development, many epithelia are formed by a mesenchymal-epithelial transition (MET). Here, we examine the major stages and underlying mechanisms of MET during blood-brain barrier formation in Drosophila We show that contact with the basal lamina is essential for the growth of the barrier-forming subperineurial glia (SPG). Septate junctions (SJs), which provide insulation of the paracellular space, are not required for MET, but are necessary for the establishment of polarized SPG membrane compartments...
January 20, 2017: Biology Open
https://www.readbyqxmd.com/read/28108377/cucurbitacin-b-purified-from-ecballium-elaterium-l-a-rich-from-tunisia-inhibits-%C3%AE-5%C3%AE-1-integrin-mediated-adhesion-migration-proliferation-of-human-glioblastoma-cell-line-and-angiogenesis
#4
Imen Touihri-Barakati, Olfa Kallech-Ziri, Wiem Ayadi, Hervé Kovacic, Belgacem Hanchi, Karim Hosni, José Luis
Integrins are essential protagonists in the complex multistep process of cancer progression and are thus attractive targets for the development of anticancer agents. Cucurbitacin B, a triterpenoid purified from the leaves of Tunisian Ecballium elaterium exhibited an anticancer effect and displayed anti-integrin activity on human glioblastoma U87 cells, without being cytotoxic at concentrations up to 500nM. Here we show that cucurbitacin B affected the adhesion and migration of U87 cells to fibronectin in a dose-dependent manner with IC50 values of 86...
January 17, 2017: European Journal of Pharmacology
https://www.readbyqxmd.com/read/28101427/dynamic-imaging-and-quantitative-analysis-of-cranial-neural-tube-closure-in-the-mouse-embryo-using-optical-coherence-tomography
#5
Shang Wang, Monica D Garcia, Andrew L Lopez, Paul A Overbeek, Kirill V Larin, Irina V Larina
Neural tube closure is a critical feature of central nervous system morphogenesis during embryonic development. Failure of this process leads to neural tube defects, one of the most common forms of human congenital defects. Although molecular and genetic studies in model organisms have provided insights into the genes and proteins that are required for normal neural tube development, complications associated with live imaging of neural tube closure in mammals limit efficient morphological analyses. Here, we report the use of optical coherence tomography (OCT) for dynamic imaging and quantitative assessment of cranial neural tube closure in live mouse embryos in culture...
January 1, 2017: Biomedical Optics Express
https://www.readbyqxmd.com/read/28092406/llfh1-mediated-interaction-between-actin-fringe-and-exocytic-vesicles-is-involved-in-pollen-tube-tip-growth
#6
Shanwei Li, Huaijian Dong, Weike Pei, Chaonan Liu, Sha Zhang, Tiantian Sun, Xiuhua Xue, Haiyun Ren
Pollen tube tip growth is an extreme form of polarized cell growth, which requires polarized exocytosis based on a dynamic actin cytoskeleton. However, the molecular basis for the connection between actin filaments and exocytic vesicles is unclear. Here, we identified a Lilium longiflorum pollen-specific formin (LlFH1) and found that it localized at the apical vesicles and plasma membrane (PM). Overexpression of LlFH1 induced excessive actin cables in the tube tip region, and downregulation of LlFH1 eliminated the actin fringe...
January 16, 2017: New Phytologist
https://www.readbyqxmd.com/read/28089989/rhoa-regulates-actin-network-dynamics-during-apical-surface-emergence-in-multiciliated-epithelial-cells
#7
Jakub Sedzinski, Edouard Hannezo, Fan Tu, Maté Biro, John B Wallingford
Homeostatic replacement of epithelial cells from basal precursors is a multistep process involving progenitor cell specification, radial intercalation and, finally, apical surface emergence. Recent data demonstrate that actin-based pushing under the control of the formin protein Fmn1 drives apical emergence in nascent multiciliated epithelial cells (MCCs), but little else is known about this actin network or the control of Fmn1. Here, we explore the role of the small GTPase RhoA in MCC apical emergence. Disruption of RhoA function reduced the rate of apical surface expansion and decreased the final size of the apical domain...
January 15, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/28087054/care-pathway-of-patients-with-metastatic-pancreatic-cancer-in-daily-practice-in-france-results-from-the-repere-national-survey
#8
Pascal Hammel, Romain Coriat, Gérard Lledo, Mariella de Bausset, Marion Selosse, Stéphane Obled, Franck Bonnetain
Data in the literature regarding the care pathway of pancreatic cancer patients are limited. The objective of the REPERE survey was to identify and describe the initial stages of the care pathway of pancreatic patients in the metastatic phase. From May to October 2015, 62 oncologists (ON) or gastroenterologists specialized in digestive oncology (GESDO) and 300 general practitioners (GP) completed an electronic questionnaire on the pathway of 728 patients recently diagnosed with metastatic pancreatic adenocarcinoma...
January 10, 2017: Bulletin du Cancer
https://www.readbyqxmd.com/read/28081137/anillin-phosphorylation-controls-timely-membrane-association-and-successful-cytokinesis
#9
Hyunjung Kim, James M Johnson, Robert F Lera, Sarang Brahma, Mark E Burkard
During cytokinesis, a contractile ring generates the constricting force to divide a cell into two daughters. This ring is composed of filamentous actin and the motor protein myosin, along with additional structural and regulatory proteins, including anillin. Anillin is a required scaffold protein that links the actomyosin ring to membrane and its organizer, RhoA. However, the molecular basis for timely action of anillin at cytokinesis remains obscure. Here, we find that phosphorylation regulates efficient recruitment of human anillin to the equatorial membrane...
January 2017: PLoS Genetics
https://www.readbyqxmd.com/read/28077222/prediction-of-relapse-after-cognitive-behavioral-treatment-of-gambling-disorder-in-individuals-with-chronic-schizophrenia-a-survival-analysis
#10
Enrique Echeburúa, Montserrat Gómez, Montserrat Freixa
Gambling disorder (GD) in individuals with chronic schizophrenia is relevant because there are higher rates of GD in schizophrenic populations (10%) than in the nonschizophrenic population (1%-5%). In addition, these patients have more severe alcohol use disorder (i.e., meeting at least 6 of the DSM-5 11 criteria for diagnosis of this disorder), higher depression scores, a poor adherence to treatment, and more frequent use of outpatient mental health care. One of the main problems in GD is therapeutic failure (defined as three or more lapse episodes during treatment) or relapse (three or more lapse episodes in the follow-up period)...
January 2017: Behavior Therapy
https://www.readbyqxmd.com/read/28077066/the-essential-requirement-of-an-animal-heme-peroxidase-protein-during-the-wing-maturation-process-in-drosophila
#11
Dondra Bailey, Mohammed Abul Basar, Sanjay Nag, Nivedita Bondhu, Shaloei Teng, Atanu Duttaroy
BACKGROUND: Thus far, a handful of genes have been shown to be related to the wing maturation process in insects. A novel heme peroxidase enzyme known as curly suppressor (Cysu)(formerly CG5873), have been characterized in this report because it is involved in wing morphogenesis. Using bioinformatics tools we found that Cysu is remarkably conserved in the genus Drosophila (>95%) as well as in invertebrates (>70%), although its vertebrate orthologs show poor homology. Time-lapse imaging and histochemical analyses have confirmed that the defective wing phenotype of Cysu is not a result of any underlying cellular alterations; instead, its wings fail to expand in mature adults...
January 11, 2017: BMC Developmental Biology
https://www.readbyqxmd.com/read/28076667/effectiveness-of-antipsychotic-drugs-for-24-month-maintenance-treatment-in-first-episode-schizophrenia-evidence-from-a-community-based-real-world-study
#12
Chen Zhang, Mei-Juan Chen, Guo-Jun Wu, Zuo-Wei Wang, Shun-Zeng Rao, Yi Zhang, Zheng-Hui Yi, Wei-Min Yang, Ke-Ming Gao, Li-Sheng Song
OBJECTIVE: Maintenance treatment of schizophrenia with antipsychotic medications has become a standard for the prevention of psychotic relapse. However, little is known about the effectiveness of antipsychotic drugs for maintenance treatment in "real-world" populations with schizophrenia. We carried out a prospective study to assess the effectiveness of the most frequently prescribed antipsychotic drugs in the maintenance treatment of schizophrenia from 2 community settings. METHODS: This study was conducted from October 2011 to December 2014...
November 2016: Journal of Clinical Psychiatry
https://www.readbyqxmd.com/read/28075574/visualization-of-mycobacterial-membrane-dynamics-in-live-cells
#13
Frances P Rodriguez-Rivera, Xiaoxue Zhou, Julie A Theriot, Carolyn R Bertozzi
Mycobacteria are endowed with a highly impermeable mycomembrane that confers intrinsic resistance to many antibiotics. Several unique mycomembrane glycolipids have been isolated and structurally characterized, but the underlying organization and dynamics of glycolipids within the cell envelope remain poorly understood. We report here a study of mycomembrane dynamics that was enabled by trehalose-fluorophore conjugates capable of labeling trehalose glycolipids in live actinomycetes. We identified fluorescein-trehalose analogs that are metabolically incorporated into the trehalose mycolates of representative Mycobacterium, Corynebacterium, Nocardia and Rhodococcus species...
January 11, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28069923/map7-regulates-axon-collateral-branch-development-in-dorsal-root-ganglia-neurons
#14
Stephen R Tymanskyj, Benjamin Yang, Aditi Falnikar, Angelo C Lepore, Le Ma
: Collateral branches from axons are key components of functional neural circuits that allow neurons to connect with multiple synaptic targets. Like axon growth and guidance, formation of collateral branches depends on the regulation of microtubules, but how such regulation is coordinated to ensure proper circuit development is not known. Based on microarray analysis, we have identified a role for microtubule-associated protein 7 (MAP7) during collateral branch development of dorsal root ganglia (DRG) sensory neurons...
January 9, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/28069186/examining-the-efficacy-of-six-published-time-lapse-imaging-embryo-selection-algorithms-to-predict-implantation-to-demonstrate-the-need-for-the-development-of-specific-in-house-morphokinetic-selection-algorithms
#15
Amy Barrie, Roy Homburg, Garry McDowell, Jeremy Brown, Charles Kingsland, Stephen Troup
OBJECTIVE: To study the efficacy of six embryo-selection algorithms (ESAs) when applied to a large, exclusive set of known implantation embryos. DESIGN: Retrospective, observational analysis. SETTING: Fertility treatment center. PATIENT(S): Women undergoing a total of 884 in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) treatment cycles (977 embryos) between September 2014 and September 2015 with embryos cultured using G-TL (Vitrolife) at 5% O2, 89% N2, 6% CO2, at 37°C in EmbryoScope instruments...
January 6, 2017: Fertility and Sterility
https://www.readbyqxmd.com/read/28068315/hyperspectral-phasor-analysis-enables-multiplexed-5d-in-vivo-imaging
#16
Francesco Cutrale, Vikas Trivedi, Le A Trinh, Chi-Li Chiu, John M Choi, Marcela S Artiga, Scott E Fraser
Time-lapse imaging of multiple labels is challenging for biological imaging as noise, photobleaching and phototoxicity compromise signal quality, while throughput can be limited by processing time. Here, we report software called Hyper-Spectral Phasors (HySP) for denoising and unmixing multiple spectrally overlapping fluorophores in a low signal-to-noise regime with fast analysis. We show that HySP enables unmixing of seven signals in time-lapse imaging of living zebrafish embryos.
January 9, 2017: Nature Methods
https://www.readbyqxmd.com/read/28065605/early-commissural-diencephalic-neurons-control-habenular-axon-extension-and-targeting
#17
Carlo A Beretta, Nicolas Dross, Luca Guglielmi, Peter Bankhead, Marina Soulika, Jose A Gutierrez-Triana, Alessio Paolini, Lucia Poggi, Julien Falk, Soojin Ryu, Marika Kapsimali, Ulrike Engel, Matthias Carl
Most neuronal populations form on both the left and right sides of the brain. Their efferent axons appear to grow synchronously along similar pathways on each side, although the neurons or their environment often differ between the two hemispheres [1-4]. How this coordination is controlled has received little attention. Frequently, neurons establish interhemispheric connections, which can function to integrate information between brain hemispheres (e.g., [5]). Such commissures form very early, suggesting their potential developmental role in coordinating ipsilateral axon navigation during embryonic development [4]...
December 29, 2016: Current Biology: CB
https://www.readbyqxmd.com/read/28064422/improved-detection-of-mineral-oil-toxicity-using-an-extended-mouse-embryo-assay
#18
Alessandra J Ainsworth, Jolene R Fredrickson, Dean E Morbeck
PURPOSE: Successful in vitro fertilization (IVF) relies on sound laboratory methods and culture conditions which depend on sensitive quality control (QC) testing. This study aimed to improve the sensitivity of mouse embryo assays (MEA) for detection of mineral oil toxicity. METHODS: Five experiments were conducted to study modifications of the standard mouse embryo assay (MEA) in order to improve sensitivity using clinical grade mineral oil with known peroxide concentrations...
January 7, 2017: Journal of Assisted Reproduction and Genetics
https://www.readbyqxmd.com/read/28064380/perioperative-factors-associated-with-hidden-blood-loss-in-intertrochanteric-fracture-patients
#19
Y Liu, Y Sun, L Fan, J Hao
BACKGROUND: The actual total blood loss after a hip fracture has been found to be considerably larger than that observed during surgery and collected in drainage postoperatively. However, no previous studies have offered reasonable explanations for the deficit between a large hemoglobin drop and a small intraoperative blood loss. The aim was to quantify blood loss associated with hip injuries and surgery, and to identify the relative risk factors for perioperative hidden blood loss. MATERIALS AND METHODS: Eighty-nine patients with a femoral intertrochanteric fracture were operated on between August 2012 and October 2013...
January 7, 2017: Musculoskeletal Surgery
https://www.readbyqxmd.com/read/28062700/intravital-imaging-of-neutrophil-recruitment-reveals-the-efficacy-of-fpr1-blockade-in-hepatic-ischemia-reperfusion-injury
#20
Masaki Honda, Takayuki Takeichi, Shintaro Hashimoto, Daiki Yoshii, Kaori Isono, Shintaro Hayashida, Yuki Ohya, Hidekazu Yamamoto, Yasuhiko Sugawara, Yukihiro Inomata
Neutrophils are considered responsible for the pathophysiological changes resulting from hepatic ischemia-reperfusion (I/R) injury, which is a complication of trauma, shock, liver resection, and transplantation. Recently, evidence is accumulating that formyl-peptide receptor (FPR) signaling constitutes an important danger signal that guides neutrophils to sites of inflammation. This study aimed to investigate dynamic neutrophil recruitment using two-photon laser-scanning microscopy (TPLSM) in response to FPR1 blockade during hepatic I/R...
January 6, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
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