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https://www.readbyqxmd.com/read/28737741/rapidly-inducible-cas9-and-dsb-ddpcr-to-probe-editing-kinetics
#2
John C Rose, Jason J Stephany, William J Valente, Bridget M Trevillian, Ha V Dang, Jason H Bielas, Dustin J Maly, Douglas M Fowler
We developed a chemically inducible Cas9 (ciCas9) and a droplet digital PCR assay for double-strand breaks (DSB-ddPCR) to investigate the kinetics of Cas9-mediated generation and repair of DSBs in cells. ciCas9 is a rapidly activated, single-component Cas9 variant engineered by replacing the protein's REC2 domain with the BCL-xL protein and fusing an interacting BH3 peptide to the C terminus. ciCas9 can be tunably activated by a compound that disrupts the BCL-xL-BH3 interaction within minutes. DSB-ddPCR demonstrates time-resolved, highly quantitative, and targeted measurement of DSBs...
July 24, 2017: Nature Methods
https://www.readbyqxmd.com/read/28737471/quality-control-by-trans-editing-factor-prevents-global-mistranslation-of-non-protein-amino-acid-%C3%AE-aminobutyrate
#3
Jo Marie Bacusmo, Alexandra B Kuzmishin, William A Cantara, Yuki Goto, Hiroaki Suga, Karin Musier-Forsyth
Accuracy in protein biosynthesis is maintained through multiple pathways, with a critical checkpoint occurring at the tRNA aminoacylation step catalyzed by aminoacyl-tRNA synthetases (ARSs). In addition to the editing functions inherent to some synthetases, single-domain trans-editing factors, which are structurally homologous to ARS editing domains, have evolved as alternative mechanisms to correct mistakes in aminoacyl-tRNA synthesis. To date, ARS-like trans-editing domains have been shown to act on specific tRNAs that are mischarged with genetically encoded amino acids...
July 24, 2017: RNA Biology
https://www.readbyqxmd.com/read/28736942/glycaemic-control-and-hypoglycaemia-in-people-with-type-2-diabetes-switching-from-twice-daily-basal-insulin-to-once-daily-insulin-glargine-300-u-ml-or-insulin-glargine-100-u-ml-edition-1-and-edition-2-subgroup-analysis
#4
Ronan Roussel, Michael C d'Emden, Miles Fisher, Francisco Javier Ampudia-Blasco, Peter Stella, Florence Bizet, Anna M G Cali, Carol H Wysham
This post hoc analysis compared glycaemic control and hypoglycaemia between insulin glargine 300 U/mL (Gla-300) and glargine 100 U/mL (Gla-100) administered once daily (QD) in people with T2DM from the EDITION 1 (basal plus mealtime insulin) and EDITION 2 (basal insulin plus oral antihyperglycaemic drugs) trials previously receiving twice-daily (BID) insulin. At randomisation, 16.9% and 20.0% of people in EDITION 1 and 2, respectively, were receiving BID basal insulin. HbA1c change from baseline to month 6 was comparable over 6 months with Gla-300 or Gla-100 (LS mean difference -0...
July 24, 2017: Diabetes, Obesity & Metabolism
https://www.readbyqxmd.com/read/28736447/prokaryotic-argonaute-proteins-novel-genome-editing-tools
#5
REVIEW
Jorrit W Hegge, Daan C Swarts, John van der Oost
Argonaute proteins constitute a highly diverse family of nucleic acid-guided proteins. They were first discovered in eukaryotes as key proteins in RNA interference systems, but homologous prokaryotic Argonaute proteins (pAgos) have also been found in archaea and bacteria. In this Progress article, we focus on long pAgo variants, a class of pAgos that are involved in nucleic acid-guided host defence against invading nucleic acids, and discuss the potential of pAgos in genome editing.
July 24, 2017: Nature Reviews. Microbiology
https://www.readbyqxmd.com/read/28736290/regulated-proteolysis-as-an-element-of-er-stress-and-autophagy-implications-for-intestinal-inflammation
#6
REVIEW
Stephanie Stengel, Berith Messner, Maren Falk-Paulsen, Nina Sommer, Philip Rosenstiel
Endoplasmic reticulum (ER) stress and autophagy are tightly controlled cellular processes, which are responsible for maintaining protein homeostasis in a cell. Impairment of the interlinking pathways have been implicated in a number of human diseases, prominently in inflammatory bowel disease, where genetic variants in several independent autophagy and ER stress related loci have been associated to increased disease risk. Autophagy is a selective quality control process, which governs the integrity of the cell by removal of aged organelles and proteins via the lysosome, but recently has been shown to actively license the outcome of other signaling pathways by guiding the proteolytic removal of signaling protein complexes (adaptophagy)...
July 20, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28735901/mechanisms-of-pannexin1-channel-gating-and-regulation
#7
REVIEW
Paige Whyte-Fagundes, Georg Zoidl
Pannexins are a family of integral membrane proteins with distinct post-translational modifications, sub-cellular localization and tissue distribution. Panx1 is the most studied and best-characterized isoform of this gene family. The ubiquitous expression, as well as its function as a major ATP release and nucleotide permeation channel, makes Panx1 a primary candidate for participating in the pathophysiology of CNS disorders. While many investigations revolve around Panx1 functions in health and disease, more recently, details started emerging about mechanisms that control Panx1 channel activity...
July 20, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28735097/protecting-the-heart-through-mk2-modulation-toward-a-role-in-diabetic-cardiomyopathy-and-lipid-metabolism
#8
REVIEW
Matthieu Ruiz, Lise Coderre, Bruce Gordon Allen, Christine Des Rosiers
Various signaling pathways have been identified in the heart as important players during development, physiological adaptation or pathological processes. This includes the MAPK families, particularly p38MAPK, which is involved in several key cellular processes, including differentiation, proliferation, apoptosis, inflammation, metabolism and survival. Disrupted p38MAPK signaling has been associated with several diseases, including cardiovascular diseases (CVD) as well as diabetes and its related complications...
July 19, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28735096/lipid-droplet-functions-beyond-energy-storage
#9
REVIEW
Michael A Welte, Alex P Gould
Lipid droplets are cytoplasmic organelles that store neutral lipids and are critically important for energy metabolism. Their function in energy storage is firmly established and increasingly well characterized. However, emerging evidence indicates that lipid droplets also play important and diverse roles in the cellular handling of lipids and proteins that may not be directly related to energy homeostasis. Lipid handling roles of droplets include the storage of hydrophobic vitamin and signaling precursors, and the management of endoplasmic reticulum and oxidative stress...
July 19, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28735095/regulation-of-lipid-droplet-associated-proteins-by-peroxisome-proliferator-activated-receptors
#10
REVIEW
Montserrat A de la Rosa Rodriguez, Sander Kersten
Excess fatty acids are stored in cells as triglycerides in specialized organelles called lipid droplets (LD). LD can be found in nearly all cell types and may expand during certain (patho)physiological conditions. The synthesis and breakdown of triglycerides and their deposition in LD is governed by a diverse set of enzymes and LD-associated proteins. These proteins serve structural roles in and around LD and regulate the activity of key lipogenic and lipolytic enzymes. The LD-associated proteins are subject to multiple regulatory mechanisms at the protein and gene expression level...
July 19, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28734978/insights-into-the-functional-properties-of-the-marneral-oxidase-cyp71a16-from-arabidopsis-thaliana
#11
Sarah Kranz-Finger, Osama Mahmoud, Esther Ricklefs, Nikolas Ditz, Patrick J Bakkes, Vlada B Urlacher
The Arabidopsis thaliana gene encoding CYP71A16 is part of the gene cluster for the biosynthesis and modification of the triterpenoid marneral. Previous investigations of A. thaliana have revealed that CYP71A16 oxidizes marneral, while it also can accept marnerol as substrate. The aim of the present study was to investigate functional properties of CYP71A16 in vitro. For this purpose, heterologous expression of a N-terminally modified version of CYP71A16 was established in Escherichia coli, which yielded up to 50mg·L(-1) recombinant enzyme...
July 19, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28734977/modulation-of-the-interaction-between-human-p450-3a4-and-b-megaterium-reductase-via-engineered-loops
#12
Silvia Castrignanò, Serena D'Avino, Giovanna Di Nardo, Gianluca Catucci, Sheila J Sadeghi, Gianfranco Gilardi
Chimerogenesis involving cytochromes P450 is a successful approach to generate catalytically self-sufficient enzymes. However, the connection between the different functional modules should allow a certain degree of flexibility in order to obtain functional and catalytically efficient proteins. We previously applied the molecular Lego approach to develop a chimeric P450 3A4 enzyme linked to the reductase domain of P450 BM3 (BMR). Three constructs were designed with the connecting loop containing no glycine, 3 glycine or 5 glycine residues and showed a different catalytic activity and coupling efficiency...
July 19, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28734966/development-of-new-ganglioside-probes-and-unraveling-of-raft-domain-structure-by-single-molecule-imaging
#13
REVIEW
Kenichi G N Suzuki, Hiromune Ando, Naoko Komura, Takahiro Fujiwara, Makoto Kiso, Akihiro Kusumi
Gangliosides are involved in a variety of biological roles and are a component of lipid rafts found in cell plasma membranes (PMs). Gangliosides are especially abundant in neuronal PMs and are essential to their physiological functions. However, the dynamic behaviors of gangliosides have not been investigated in living cells due to a lack of fluorescent probes that behave like their parental molecules. We have recently developed, using an entirely chemical method, four new ganglioside probes (GM1, GM2, GM3, and GD1b) that act similarly to their parental molecules in terms of raft partitioning and binding affinity...
July 19, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28734794/botulinum-toxin-a-is-effective-to-treat-tension-type-headache-caused-by-hemifacial-spasm
#14
Atsushi Mizuma, Eiichiro Nagata, Takashi Yasuda, Maiko Kouchi, Taira Nakayama, Kazunari Honma, Kentaro Tokuoka, Yasuhisa Kitagawa, Shigeru Nogawa, Shunya Takizawa
OBJECTIVE: We examined the relationship between hemifacial spasm (HFS; a form of cranio-cervical dystonia) and chronic primary headache, including tension-type headache (TTH). We also examined whether botulinum toxin A (BoNT/A) therapy for HFS ameliorates concomitant TTH. METHODS: Fifty-one HFS patients receiving BoNT/A therapy were recruited. Patients' characteristics (including age, gender, chronic headache history, exercise habits, stiff neck, cervical spondylolysis history), stress factors, worsening/new onset of headache associated with HFS, and dose of BoNT/A were examined...
July 19, 2017: Journal of Clinical Neuroscience: Official Journal of the Neurosurgical Society of Australasia
https://www.readbyqxmd.com/read/28734360/revealing-the-biography-of-a-hidden-medieval-manuscript-using-synchrotron-and-conventional-imaging-techniques
#15
E Pouyet, S Devine, T Grafakos, R Kieckhefer, J Salvant, L Smieska, A Woll, A Katsaggelos, O Cossairt, M Walton
Reading the content of hidden texts from ancient manuscripts has become an increasingly important endeavor thanks to the variety of non-destructive analytical tools and image processing routines available for this task. In this study, portable macro X-Ray Fluorescence (MA-XRF-tube), Visible Hyperspectral Imaging (HSI) together with Synchrotron based macro X-Ray Fluorescence (MA-XRF-SR) are combined with signal processing methods to reveal the biography of a degraded manuscript recycled as binding material for a 16th century printed edition of Hesiod's Works and Days...
August 22, 2017: Analytica Chimica Acta
https://www.readbyqxmd.com/read/28734060/effects-of-eddy-currents-on-selective-spectral-editing-experiments-at-3t
#16
Georg Oeltzschner, Karim Snoussi, Nicolaas A Puts, Mark Mikkelsen, Ashley D Harris, Subechhya Pradhan, Kyrana Tsapkini, Michael Schär, Peter B Barker, Richard A E Edden
PURPOSE: To investigate frequency-offset effects in edited magnetic resonance spectroscopy (MRS) experiments arising from B0 eddy currents. MATERIALS AND METHODS: Macromolecule-suppressed (MM-suppressed) γ-aminobutyric acid (GABA)-edited experiments were performed at 3T. Saturation-offset series of MEGA-PRESS experiments were performed in phantoms, in order to investigate different aspects of the relationship between the effective editing frequencies and eddy currents associated with gradient pulses in the sequence...
July 22, 2017: Journal of Magnetic Resonance Imaging: JMRI
https://www.readbyqxmd.com/read/28733609/suppression-of-hbv-replication-by-the-expression-of-nickase-and-nuclease-dead-cas9
#17
Takeshi Kurihara, Takasuke Fukuhara, Chikako Ono, Satomi Yamamoto, Kentaro Uemura, Toru Okamoto, Masaya Sugiyama, Daisuke Motooka, Shota Nakamura, Masato Ikawa, Masashi Mizokami, Yoshihiko Maehara, Yoshiharu Matsuura
Complete removal of hepatitis B virus (HBV) DNA from nuclei is difficult by the current therapies. Recent reports have shown that a novel genome-editing tool using Cas9 with a single-guide RNA (sgRNA) system can cleave the HBV genome in vitro and in vivo. However, induction of a double-strand break (DSB) on the targeted genome by Cas9 risks undesirable off-target cleavage on the host genome. Nickase-Cas9 cleaves a single strand of DNA, and thereby two sgRNAs are required for inducing DSBs. To avoid Cas9-induced off-target mutagenesis, we examined the effects of the expressions of nickase-Cas9 and nuclease dead Cas9 (d-Cas9) with sgRNAs on HBV replication...
July 21, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28733196/proteasome-subunit-beta-type-1-p11a-polymorphism-is-a-new-prognostic-marker-in-multiple-myeloma
#18
Gergely Varga, Gábor Mikala, Katalin Piroska Kiss, Éva Kosóczki, Edit Szabó, Nóra Meggyesi, Katalin Balassa, Petra Kövy, Bálint Tegze, Gergely Szombath, Attila Tordai, Hajnalka Andrikovics, László Homolya, Tamás Masszi
BACKGROUND: Proteasome subunit beta type 1 (PSMB1) rs12717 polymorphism, a single nucleotide polymorphism with unknown functional effect, was recently reported to influence response to bortezomib-based therapy in follicular lymphoma. PATIENTS AND METHODS: We retrospectively analyzed the prognostic impact of this polymorphism in 211 consecutively diagnosed multiple myeloma cases, and performed in vitro experiments to look into its functional consequences. RESULTS: On univariate analysis, patients carrying the variant G allele showed significantly shorter progression-free survival (PFS) with a pattern suggestive of a gene-dose effect (PFS 26...
June 30, 2017: Clinical Lymphoma, Myeloma & Leukemia
https://www.readbyqxmd.com/read/28732771/application-of-the-gene-editing-tool-crispr-cas9-for-treating-neurodegenerative-diseases
#19
REVIEW
Nivya Kolli, Ming Lu, Panchanan Maiti, Julien Rossignol, Gary Leo Dunbar
Increased accumulation of transcribed protein from the damaged DNA and reduced DNA repair capability contributes to numerous neurological diseases for which effective treatments are lacking. Gene editing techniques provide new hope for replacing defective genes and DNA associated with neurological diseases. With advancements in using such editing tools as zinc finger nucleases (ZFNs), meganucleases, and transcription activator-like effector nucleases (TALENs), etc., scientists are able to design DNA-binding proteins, which can make precise double-strand breaks (DSBs) at the target DNA...
July 18, 2017: Neurochemistry International
https://www.readbyqxmd.com/read/28732551/novel-additions-to-the-ajcc-s-new-staging-systems-for-skin-cancer
#20
J Cañueto, C Román-Curto
The eighth edition of the staging manual of the American Joint Committee on Cancer incorporates important changes in the classification of skin cancers. Coming 40 years after the first edition, the latest manual preserves its specific system for Merkel cell carcinoma and takes into account recent publications on the prognosis of squamous cell carcinoma by defining a completely new T category for this neoplasm. Staging for squamous cell carcinoma considers head and neck tumors (excluding the eyelid) and does not offer solutions for other sites except the vulva, penis, and perianal region...
July 18, 2017: Actas Dermo-sifiliográficas
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