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https://www.readbyqxmd.com/read/28533524/enhancing-the-genome-editing-toolbox-genome-wide-crispr-arrayed-libraries
#1
Emmanouil Metzakopian, Alex Strong, Vivek Iyer, Alex Hodgkins, Konstantinos Tzelepis, Liliana Antunes, Mathias J Friedrich, Qiaohua Kang, Teresa Davidson, Jacob Lamberth, Christina Hoffmann, Gregory D Davis, George S Vassiliou, William C Skarnes, Allan Bradley
CRISPR-Cas9 technology has accelerated biological research becoming routine for many laboratories. It is rapidly replacing conventional gene editing techniques and has high utility for both genome-wide and gene-focussed applications. Here we present the first individually cloned CRISPR-Cas9 genome wide arrayed sgRNA libraries covering 17,166 human and 20,430 mouse genes at a complexity of 34,332 sgRNAs for human and 40,860 sgRNAs for the mouse genome. For flexibility in generating stable cell lines the sgRNAs have been cloned in a lentivirus backbone containing PiggyBac transposase recognition elements together with fluorescent and drug selection markers...
May 22, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28532483/naturally-acquired-humoral-and-cellular-immune-responses-to-plasmodium-vivax-merozoite-surface-protein-8-in-patients-with-p-vivax-infection
#2
Yang Cheng, Bo Wang, Siriruk Changrob, Jin-Hee Han, Jetsumon Sattabongkot, Kwon-Soo Ha, Patchanee Chootong, Feng Lu, Jun Cao, Myat Htut Nyunt, Won Sun Park, Seok-Ho Hong, Chae Seung Lim, Takafumi Tsuboi, Eun-Taek Han
BACKGROUND: Thirty-one glycosylphosphatidylinositol (GPI)-anchored proteins of Plasmodium vivax, merozoite surface protein 1 (MSP1), MSP1 paralogue, MSP4, MSP5, MSP8, and MSP10 have been reported from homologs of Plasmodium falciparum by gene annotation with bioinformatics tools. These GPI-anchored proteins contain two epidermal growth factor (EGF)-like domains at its C-terminus. Here, P. vivax merozoite surface protein 8 (PvMSP8) are considered as potential targets of protective immunity...
May 22, 2017: Malaria Journal
https://www.readbyqxmd.com/read/28531887/expression-and-characterization-of-membrane-type-4-matrix-metalloproteinase-mt4-mmp-and-its-different-forms-in-melanoma
#3
Bettina Hieronimus, Julian Pfohl, Christian Busch, Lutz Graeve
BACKGROUND/AIMS: Membrane-type matrix metalloproteinases (MT-MMPs) are expressed on the cell surface and hydrolyze extracellular matrix components and signaling molecules by which they influence cancer cell migration and metastasis. Two of the six known MT-MMPs are anchored to the plasma membrane via a GPI anchor, one of which is MT4-MMP. Only little is known about MT4-MMP expression, synthesis, regulation and degradation. METHODS: We analyzed several human cancer cell lines as well as tissue homogenates using Western blotting and quantitative PCR for the expression of MT4-MMP...
May 25, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28526096/pvgama-reticulocyte-binding-activity-predicting-conserved-functional-regions-by-natural-selection-analysis
#4
Luis A Baquero, Darwin A Moreno-Pérez, Diego Garzón-Ospina, Johanna Forero-Rodríguez, Heidy D Ortiz-Suárez, Manuel A Patarroyo
BACKGROUND: Adhesin proteins are used by Plasmodium parasites to bind and invade target cells. Hence, characterising molecules that participate in reticulocyte interaction is key to understanding the molecular basis of Plasmodium vivax invasion. This study focused on predicting functionally restricted regions of the P. vivax GPI-anchored micronemal antigen (PvGAMA) and characterising their reticulocyte binding activity. RESULTS: The pvgama gene was initially found in P...
May 19, 2017: Parasites & Vectors
https://www.readbyqxmd.com/read/28518050/acetylcholinesterase-provides-new-insights-into-red-blood-cell-ageing-in-vivo-and-in-vitro
#5
Joames K Freitas Leal, Merel J W Adjobo-Hermans, Roland Brock, Giel J C G M Bosman
BACKGROUND: During its 120 days sojourn in the circulation, the red blood cell (RBC) remodels its membrane. Acetylcholinesterase (AChE) is a glycosylphosphatidylinositol (GPI)-linked enzyme that may serve as a marker for membrane processes occurring this ageing-associated remodelling process. MATERIALS AND METHODS: Expression and enzymatic activity of AChE were determined on RBCs of various ages, as obtained by separation based on volume and density (ageing in vivo), and on RBCs of various times of storage in blood bank conditions (ageing in vitro), as well as on RBC-derived vesicles...
May 2017: Blood Transfusion, Trasfusione del Sangue
https://www.readbyqxmd.com/read/28516949/paroxysmal-nocturnal-haemoglobinuria
#6
REVIEW
Anita Hill, Amy E DeZern, Taroh Kinoshita, Robert A Brodsky
Paroxysmal nocturnal haemoglobinuria (PNH) is a clonal haematopoietic stem cell (HSC) disease that presents with haemolytic anaemia, thrombosis and smooth muscle dystonias, as well as bone marrow failure in some cases. PNH is caused by somatic mutations in PIGA (which encodes phosphatidylinositol N-acetylglucosaminyltransferase subunit A) in one or more HSC clones. The gene product of PIGA is required for the biosynthesis of glycosylphosphatidylinositol (GPI) anchors; thus, PIGA mutations lead to a deficiency of GPI-anchored proteins, such as complement decay-accelerating factor (also known as CD55) and CD59 glycoprotein (CD59), which are both complement inhibitors...
May 18, 2017: Nature Reviews. Disease Primers
https://www.readbyqxmd.com/read/28510056/the-two-sides-of-a-lipid-protein-story
#7
REVIEW
Luis G Mansor Basso, Luis F Santos Mendes, Antonio J Costa-Filho
Protein-membrane interactions play essential roles in a variety of cell functions such as signaling, membrane trafficking, and transport. Membrane-recruited cytosolic proteins that interact transiently and interfacially with lipid bilayers perform several of those functions. Experimental techniques capable of probing changes on the structural dynamics of this weak association are surprisingly limited. Among such techniques, electron spin resonance (ESR) has the enormous advantage of providing valuable local information from both membrane and protein perspectives by using intrinsic paramagnetic probes in metalloproteins or by attaching nitroxide spin labels to proteins and lipids...
June 2016: Biophysical Reviews
https://www.readbyqxmd.com/read/28507137/arabidopsis-glycosylphosphatidylinositol-anchored-protein-llg1-associates-with-and-modulates-fls2-to-regulate-innate-immunity
#8
Qiujing Shen, Gildas Bourdais, Huairong Pan, Silke Robatzek, Dingzhong Tang
Plants detect and respond to pathogen invasion with membrane-localized pattern recognition receptors (PRRs), which recognize pathogen-associated molecular patterns (PAMPs) and activate downstream immune responses. Here we report that Arabidopsis thaliana LORELEI-LIKE GPI-ANCHORED PROTEIN 1 (LLG1), a coreceptor of the receptor-like kinase FERONIA, regulates PRR signaling. In a forward genetic screen for suppressors of enhanced disease resistance 1 (edr1), we identified the point mutation llg1-3, which suppresses edr1 disease resistance but does not affect plant growth and development...
May 15, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28507000/the-cell-wall-localized-atypical-%C3%AE-1-3-glucanase-zerzaust-controls-tissue-morphogenesis-in-arabidopsis-thaliana
#9
Prasad Vaddepalli, Lynette Fulton, Jennifer Wieland, Katrin Wassmer, Milena Schaeffer, Stefanie Ranf, Kay Schneitz
Orchestration of cellular behavior in plant organogenesis requires integration of intercellular communication and cell wall dynamics. The underlying signaling mechanisms are poorly understood. Tissue morphogenesis in Arabidopsis depends on the receptor-like kinase STRUBBELIG. Mutations in ZERZAUST were previously shown to result in a strubbelig-like mutant phenotype. Here we report on the molecular identification and functional characterization of ZERZAUST We show that ZERZAUST encodes a putative GPI-anchored β-1,3 glucanase suggested to degrade the cell wall polymer callose...
May 15, 2017: Development
https://www.readbyqxmd.com/read/28500877/clinically-isolated-aortitis-pitfalls-progress-and-possibilities
#10
REVIEW
Ilkay Cinar, He Wang, James R Stone
Non-infectious aortitis may be caused by several distinct systemic rheumatologic diseases. In some patients, aortitis is identified either pathologically or radiologically in the absence of clinical evidence of a systemic vasculitis. By consensus nomenclature, such cases are referred to as clinically isolated aortitis (CIA). Some systemic disorders may initially present as CIA including giant cell arteritis (GCA), IgG4-related disease, infectious aortitis, and granulomatosis with polyangiitis. CIA most commonly occurs in women of European descent over the age of 50 and, thus, mirrors the gender, age, and geographic distribution of GCA...
April 23, 2017: Cardiovascular Pathology: the Official Journal of the Society for Cardiovascular Pathology
https://www.readbyqxmd.com/read/28497740/-the-pig-a-gene-as-a-new-biomarker-of-mutagenesis-proof-of-concept-and-technical-specifications
#11
Pierre Castel, Xavier Carcopino, Stéphane Robert, Rémi Bonetto, Didier Cowen, Thierry Orsiere
Gene mutations are not directly detected by current genotoxicity assays and most of them need a cell culture step. The whole blood PIG-A assay consists in the detection of the mutation frequency within the PIG-A sentinel gene by identification of glycosyl-phosphatidyl-inositol (GPI-) deficient cells. PIG-A mutated/GPI-deficient cells can be detected by flow cytometry as they no longer express surface fluorescence for GPI-linked markers. The last researches have focused on cell enrichment techniques leading to increased throughput and sensitivity...
April 2017: Médecine Sciences: M/S
https://www.readbyqxmd.com/read/28495885/proteolytic-cleavage-of-the-hydrophobic-domain-in-the-cav%C3%AE-2%C3%AE-1-subunit-improves-assembly-and-activity-of-cardiac-cav1-2-channels
#12
Emilie Segura, Benoîte Bourdin, Marie-Philippe Tétreault, Julie Briot, Bruce G Allen, Gaétan Mayer, Lucie Parent
Voltage-gated L-type CaV1.2 channels in cardiomyocytes exist as heteromeric complexes with the pore-forming CaVα1, CaVβ, and CaVα2δ1 subunits. The full complement of subunits is required to reconstitute the native-like properties of L-type Ca(2+) currents but the molecular determinants responsible for the formation of the heteromeric complex are still being studied. Enzymatic treatment with PI-PLC, a phospholipase C specific for the cleavage of glycosyl-phosphatidylinositol (GPI)-anchored proteins, disrupted plasma membrane localization of the cardiac CaVα2δ1 prompting us to investigate deletions of its hydrophobic transmembrane domain...
May 11, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28491942/a-dataset-of-multi-contrast-population-averaged-brain-mri-atlases-of-a-parkinson-s-disease-cohort
#13
Yiming Xiao, Vladimir Fonov, M Mallar Chakravarty, Silvain Beriault, Fahd Al Subaie, Abbas Sadikot, G Bruce Pike, Gilles Bertrand, D Louis Collins
Parkinson׳s disease (PD) is a neurodegenerative disease that primarily affects the motor functions of the patients. Research and surgical treatment of PD (e.g., deep brain stimulation) often require human brain atlases for structural identification or as references for anatomical normalization. However, two pitfalls exist for many current atlases used for PD. First, most atlases do not represent the disease-specific anatomy as they are based on healthy young subjects. Second, subcortical structures, such as the subthalamic nucleus (STN) used in deep brain stimulation procedures, are often not well visualized...
June 2017: Data in Brief
https://www.readbyqxmd.com/read/28490634/inflammatory-cytokines-down-regulate-the-barrier-protective-prostasin-matriptase-proteolytic-cascade-early-in-experimental-colitis
#14
Marguerite S Buzza, Tierra A Johnson, Gregory D Conway, Erik W Martin, Subhradip Mukhopadhyay, Terez Shea-Donohue, Toni M Antalis
Compromised gastrointestinal barrier function is strongly associated with the progressive and destructive pathologies of the two main forms of IBD, Ulcerative Colitis (UC) and Crohns disease (CD). Matriptase is a membrane-anchored serine protease encoded by Suppression of Tumorigenicity-14 (ST14) gene, which is critical for epithelial barrier development and homeostasis. Matriptase barrier protective activity is linked with the glycosyl-phosphatidyl-inositol (GPI)-anchored serine protease prostasin, which is a co-factor for matriptase zymogen activation...
May 10, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28486994/the-emergence-of-a-novel-sequence-type-of-mdr-acinetobacter-baumannii-from-the-intensive-care-unit-of-an-egyptian-tertiary-care-hospital
#15
Doaa Mohammad Ghaith, Mai Mahmoud Zafer, Mohamed Hamed Al-Agamy, Essam J Alyamani, Rayan Y Booq, Omar Almoazzamy
BACKGROUND AND AIM OF WORK: Acinetobacter baumannii is known for nosocomial outbreaks worldwide. In this study, we aimed to investigate the antibiotic susceptibility patterns and the clonal relationship of A. baumannii isolates from the intensive care unit (ICU) of an Egyptian hospital. METHODS: In the present study, 50 clinical isolates of multidrug resistant (MDR)-A. baumannii were obtained from patients admitted into the ICU from June to December 2015. All isolates were analyzed for antimicrobial susceptibilities...
May 10, 2017: Annals of Clinical Microbiology and Antimicrobials
https://www.readbyqxmd.com/read/28484941/proteomics-as-a-tool-to-identify-new-targets-against-aspergillus-and-scedosporium-in-the-context-of-cystic-fibrosis
#16
Andoni Ramirez-Garcia, Aize Pellon, Idoia Buldain, Aitziber Antoran, Aitana Arbizu-Delgado, Xabier Guruceaga, Aitor Rementeria, Fernando L Hernando
Cystic fibrosis (CF) is a genetic disorder that increases the risk of suffering microbial, including fungal, infections. In this paper, proteomics-based information was collated relating to secreted and cell wall proteins with potential medical applications from the most common filamentous fungi in CF, i.e., Aspergillus and Scedosporium/Lomentospora species. Among the Aspergillus fumigatus secreted allergens, β-1,3-endoglucanase, the alkaline protease 1 (Alp1/oryzin), Asp f 2, Asp f 13/15, chitinase, chitosanase, dipeptidyl-peptidase V (DppV), the metalloprotease Asp f 5, mitogillin/Asp f 1, and thioredoxin reductase receive a special mention...
May 8, 2017: Mycopathologia
https://www.readbyqxmd.com/read/28484166/molecular-genetics-biochemistry-and-biology-of-pnh
#17
Taroh Kinoshita
Paroxysmal nocturnal hemoglobinuria (PNH) manifests by clonal expansion of mutant hematopoietic stem cells (HSCs) bearing a somatic mutation in the X-linked PIGA gene. PIGA mutations cause defective biosynthesis of GPI and cell surface deficiency of GPI-anchored proteins such as DAF and CD59, leading to intravascular hemolysis and thrombosis. These two major symptoms of PNH can be controlled by eculizumab, an anti-C5 monoclonal antibody. Bone marrow failure, the third major symptom of PNH, is autoimmune-mediated and contributes to the clonal expansion of GPI-defective HSCs by selectively attacking GPI-positive wild-type HSCs...
2017: [Rinshō Ketsueki] the Japanese Journal of Clinical Hematology
https://www.readbyqxmd.com/read/28482115/arabidopsis-thaliana-fla4-functions-as-a-glycan-stabilized-soluble-factor-via-its-carboxy-proximal-fasciclin-1-domain
#18
Hui Xue, Christiane Veit, Lindy Abas, Theodora Tryfona, Daniel Maresch, Martiniano M Ricardi, Jose M Estevez, Richard Strasser, Georg J Seifert
Fasciclin like arabinogalactan proteins (FLAs) are involved in numerous important functions in plants but the relevance of their complex structure to physiological function and cellular fate is unresolved. Using a fully functional fluorescent version of Arabidopsis thaliana FLA4 we show that this protein is localized at the plasma membrane as well as in endosomes and soluble in the apoplast. FLA4 is likely to be GPI-anchored, is highly N-glycosylated and carries two O-glycan epitopes previously associated with AGPs...
May 8, 2017: Plant Journal: for Cell and Molecular Biology
https://www.readbyqxmd.com/read/28479586/comparison-of-pallidal-and-subthalamic-deep-brain-stimulation-in-parkinson-s-disease-therapeutic-and-adverse-effects
#19
Ho-Sung Ryu, Mi-Sun Kim, Sooyeoun You, Mi-Jung Kim, Young Jin Kim, Juyeon Kim, Kiju Kim, Sun Ju Chung
OBJECTIVE: To compare the therapeutic and adverse effects of globus pallidus interna (GPi) and subthalamic nucleus (STN) deep brain stimulation (DBS) for the treatment of advanced Parkinson's disease (PD). METHODS: We retrospectively analyzed the clinical data of patients with PD who underwent GPi (n = 14) or STN (n = 28) DBS surgery between April 2002 and May 2014. The subjects were matched for age at surgery and disease duration. The Unified Parkinson's Disease Rating Scale (UPDRS) scores and levodopa equivalent dose (LED) at baseline and 12 months after surgery were used to assess the therapeutic effects of DBS...
May 2017: Journal of Movement Disorders
https://www.readbyqxmd.com/read/28475061/modulation-of-parkinsonian-state-with-uncertain-disturbance-based-on-sliding-mode-control
#20
Yulin Zhu, Jiang Wang, Huiyan Li, Bin Deng, Chen Liu
Parkinson's disease (PD) is a degenerative disorder of central nervous system that endangers the olds' health seriously. The motor symptoms of PD can be attributed to the distorted relay reliability of thalamus to cortical sensorimotor input that results from the increase of inhibitory input from internal segment of the globus pallidum (GPi). Based on this, we construct the GPithalamocortical computational model to generate the normal and pathological firing patterns by varying GPi spike train input. A kind of closed-loop deep brain stimulation (DBS) strategy is proposed here...
May 2, 2017: IEEE Transactions on Neural Systems and Rehabilitation Engineering
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