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https://www.readbyqxmd.com/read/29353442/flexible-pseudotyping-of-retrovirus-using-recombinase-mediated-cassette-exchange
#1
Hugo R Soares, Ana I Almeida, Hélio A Tomás, Paula M Alves, Ana S Coroadinha
OBJECTIVE: Develop an engineered cell line containing two flexible gene expression systems enabling the continuous production of tailor-made recombinant gammaretrovirus with predictable productivities through targeted integration. RESULTS: Dual-FLEX cells (dFLEX) contain two independent recombinase-mediated cassette exchange (RMCE) systems which confer flexibility to the expression of different transgene and envelope combinations. The flexible envelope expression in dFLEX cells was validated by pseudotyping retrovirus particles with three different viral envelope proteins-GaLV, 4070A and VSV-G...
January 20, 2018: Biotechnology Letters
https://www.readbyqxmd.com/read/29351814/setdb1-prevents-tet2-dependent-activation-of-iap-retroelements-in-na%C3%A3-ve-embryonic-stem-cells
#2
Özgen Deniz, Lorenzo de la Rica, Kevin C L Cheng, Dominik Spensberger, Miguel R Branco
BACKGROUND: Endogenous retroviruses (ERVs), which are responsible for 10% of spontaneous mouse mutations, are kept under control via several epigenetic mechanisms. The H3K9 histone methyltransferase SETDB1 is essential for ERV repression in embryonic stem cells (ESCs), with DNA methylation also playing an important role. It has been suggested that SETDB1 protects ERVs from TET-dependent DNA demethylation, but the relevance of this mechanism for ERV expression remains unclear. Moreover, previous studies have been performed in primed ESCs, which are not epigenetically or transcriptionally representative of preimplantation embryos...
January 19, 2018: Genome Biology
https://www.readbyqxmd.com/read/29351216/in-silico-and-in-vitro-study-of-the-bromelain-phytochemical-complex-inhibition-of-phospholipase-a2-pla2
#3
Fatahiya Mohamed Tap, Fadzilah Adibah Abd Majid, Hassan Fahmi Ismail, Tet Soon Wong, Kamyar Shameli, Mikio Miyake, Nurul Bahiyah Ahmad Khairudin
Phospholipase A2 (Pla2) is an enzyme that induces inflammation, making Pla2 activity an effective approach to reduce inflammation. Therefore, investigating natural compounds for this Pla2 inhibitory activity has important therapeutic potential. The objective of this study was to investigate the potential in bromelain-phytochemical complex inhibitors via a combination of in silico and in vitro methods. Bromelain-amenthoflavone displays antagonistic effects on Pla2. Bromelian-asiaticoside and bromelain-diosgenin displayed synergistic effects at high concentrations of the combined compounds, with inhibition percentages of more than 70% and 90%, respectively, and antagonistic effects at low concentrations...
January 19, 2018: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/29350135/plasmid-encoded-transferable-mecb-mediated-methicillin-resistance-in-staphylococcus-aureus
#4
Karsten Becker, Sarah van Alen, Evgeny A Idelevich, Nina Schleimer, Jochen Seggewiß, Alexander Mellmann, Ursula Kaspar, Georg Peters
During cefoxitin-based nasal screening, phenotypically categorized methicillin-resistant Staphylococcus aureus (MRSA) was isolated and tested negative for the presence of the mecA and mecC genes as well as for the SCCmec-orfX junction region. The isolate was found to carry a mecB gene previously described for Macrococcus caseolyticus but not for staphylococcal species. The gene is flanked by β-lactam regulatory genes similar to mecR, mecI, and blaZ and is part of an 84.6-kb multidrug-resistance plasmid that harbors genes encoding additional resistances to aminoglycosides (aacA-aphD, aphA, and aadK) as well as macrolides (ermB) and tetracyclines (tetS)...
February 2018: Emerging Infectious Diseases
https://www.readbyqxmd.com/read/29345269/tributyltin-induces-epigenetic-changes-and-decreases-the-expression-of-nuclear-respiratory-factor-1
#5
Saki Hanaoka, Keishi Ishida, Saki Tanaka, Shuichiro Sakamoto, Katsuhiro Okuda, Seigo Sanoh, Shigeru Ohta, Yaichiro Kotake
Tributyltin (TBT), a common organotin environmental pollutant, has been widely used as a component of marine antifouling paints. We previously reported that exposure to TBT inhibits the expression and DNA binding of nuclear respiratory factor-1 (NRF-1) and causes neurotoxicity. In the present study, we focused on the epigenetic effects of TBT and investigated whether TBT decreases NRF-1 expression via epigenetic modifications in SH-SY5Y human neuroblastoma cells. First, we found that exposure to 300 nM TBT decreases NRF-1 expression...
January 18, 2018: Metallomics: Integrated Biometal Science
https://www.readbyqxmd.com/read/29339836/the-role-of-metabolic-enzymes-in-mesenchymal-tumors-and-tumor-syndromes-genetics-pathology-and-molecular-mechanisms
#6
REVIEW
Inga-Marie Schaefer, Jason L Hornick, Judith V M G Bovée
The discovery of mutations in genes encoding the metabolic enzymes isocitrate dehydrogenase (IDH), succinate dehydrogenase (SDH), and fumarate hydratase (FH) has expanded our understanding not only of altered metabolic pathways but also epigenetic dysregulation in cancer. IDH1/2 mutations occur in enchondromas and chondrosarcomas in patients with the non-hereditary enchondromatosis syndromes Ollier disease and Maffucci syndrome and in sporadic tumors. IDH1/2 mutations result in excess production of the oncometabolite (D)-2-hydroxyglutarate...
January 16, 2018: Laboratory Investigation; a Journal of Technical Methods and Pathology
https://www.readbyqxmd.com/read/29339137/oxygen-induced-alterations-in-the-expression-of-chromatin-modifying-enzymes-and-the-transcriptional-regulation-of-imprinted-genes
#7
William M Skiles, Avery Kester, Jane H Pryor, Mark E Westhusin, Michael C Golding, Charles R Long
Embryo culture and assisted reproductive technologies have been associated with a disproportionately high number of epigenetic abnormalities in the resulting offspring. However, the mechanisms by which these techniques influence the epigenome remain poorly defined. In this study, we evaluated the capacity of oxygen concentration to influence the transcriptional control of a selection of key enzymes regulating chromatin structure. In mouse embryonic stem cells, oxygen concentrations modulated the transcriptional regulation of the TET family of enzymes, as well as the de novo methyltransferase Dnmt3a...
January 12, 2018: Gene Expression Patterns: GEP
https://www.readbyqxmd.com/read/29331106/kbtbd2-inhibits-the-cytotoxic-activity-of-immortalized-nk-cells-through-down-regulating-mtor-signaling-in-a-mouse-hepatocellular-carcinoma-model
#8
Kai Dai, Yabing Huang, Zubing Chen, Xiaomei Sun, Lihua Yang, Yingan Jiang
Natural killer cell (NK cell)-based immunotherapy is a promising therapeutic strategy for hepatocellular carcinoma (HCC). However, the molecular mechanisms underlying the regulation of NK cell function in the tumor sites are not completely elucidated. In this study, we identified the enhanced expression of kelch repeat and BTB (POZ) domain containing 2 (Kbtbd2) in intratumoral NK cells in a mouse HCC implantation model as a negative regulator of NK cells. To investigate this interaction,, we used a Tet-on inducible expression system to control Kbtbd2 expression in an immortalized mouse NK cell line KIL C...
January 13, 2018: European Journal of Immunology
https://www.readbyqxmd.com/read/29329231/development-of-a-cytocompatible-scaffold-from-pig-immature-testicular-tissue-allowing-human-sertoli-cell-attachment-proliferation-and-functionality
#9
Maxime Vermeulen, Federico Del Vento, Francesca de Michele, Jonathan Poels, Christine Wyns
Cryopreservation of immature testicular tissue before chemo/radiotherapy is the only option to preserve fertility of cancer-affected prepubertal boys. To avoid reintroduction of malignant cells, development of a transplantable scaffold by decellularization of pig immature testicular tissue (ITT) able to support decontaminated testicular cells could be an option for fertility restoration in these patients. We, therefore, compared decellularization protocols to produce a cytocompatible scaffold. Fragments of ITT from 15 piglets were decellularized using three protocols: sodium dodecyl sulfate (SDS)-Triton (ST), Triton-SDS-Triton (TST) and trypsin 0...
January 12, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29325442/pooled-generation-of-lentiviral-tetracycline-regulated-microrna-embedded-short-hairpin-rna-libraries
#10
Felix Ferdinand Adams, Thomas Hoffmann, Johannes Zuber, Dirk Heckl, Axel Schambach, Adrian Schwarzer
Short hairpin RNA (shRNA) screens are powerful tools to probe genetic dependencies in loss-of-function studies, like the identification of therapeutic targets in cancer research. Lentivirally delivered shRNAs embedded in endogenous microRNA contexts (shRNAmiRs) mediate efficient long-term suppression of target genes suitable for numerous experimental contexts and clinical applications. Here, we describe an easy-to-use laboratory protocol covering the design and pooled assembly of focused shRNAmiR libraries into an optimized, Tet-inducible all-in-one lentiviral vector, packaging of viral particles, followed by retrieval and quantification of hairpin sequences after cellular DNA-recovery...
January 12, 2018: Human Gene Therapy Methods
https://www.readbyqxmd.com/read/29324752/tet-protein-function-during-drosophila-development
#11
Fei Wang, Svetlana Minakhina, Hiep Tran, Neha Changela, Joseph Kramer, Ruth Steward
The TET (Ten-eleven translocation) 1, 2 and 3 proteins have been shown to function as DNA hydroxymethylases in vertebrates and their requirements have been documented extensively. Recently, the Tet proteins have been shown to also hydroxylate 5-methylcytosine in RNA. 5-hydroxymethylcytosine (5hmrC) is enriched in messenger RNA but the function of this modification has yet to be elucidated. Because Cytosine methylation in DNA is barely detectable in Drosophila, it serves as an ideal model to study the biological function of 5hmrC...
2018: PloS One
https://www.readbyqxmd.com/read/29323188/a-tet-off-gene-expression-system-for-validation-of-antifungal-drug-targets-in-a-murine-invasive-pulmonary-aspergillosis-model
#12
Yutian Peng, Hua Zhang, Min Xu, Man-Wah Tan
Aspergillus fumigatus is one of the major causes of invasive pulmonary aspergillosis in immunocompromised patients. Novel antifungal therapy is in urgent need due to emerging resistance and adverse toxicity of current antifungal drugs. Gene products that are essential for Aspergillus viability during infection are attractive drug targets. To characterize these genes in vivo we developed a Tet-Off gene expression system in A. fumigatus, whereby the administration of doxycycline resulted in down regulation of the gene whose expression is under the control of the Tet-Off promoter...
January 11, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29315376/autologous-cd34-cell-therapy-improves-exercise-capacity-angina-frequency-and-reduces-mortality-in-no-option-refractory-angina-a-patient-level-pooled-analysis-of-randomized-double-blinded-trials
#13
Timothy D Henry, Douglas W Losordo, Jay H Traverse, Richard A Schatz, E Marc Jolicoeur, Gary L Schaer, Robert Clare, Karen Chiswell, Christopher J White, F David Fortuin, Dean J Kereiakes, Andreas M Zeiher, Warren Sherman, Andrea S Hunt, Thomas J Povsic
Aims: Autologous CD34+ (auto-CD34+) cells represent an attractive option for the treatment of refractory angina. Three double-blinded randomized trials (n = 304) compared intramyocardial (IM) auto-CD34+ cells with IM placebo injections to affect total exercise time (TET), angina frequency (AF), and major adverse cardiac events (MACE). Patient-level data were pooled from the Phase I, Phase II ACT-34, ACT-34 extension, and Phase III RENEW trials to determine the efficacy and safety of auto-CD34+ cells...
January 5, 2018: European Heart Journal
https://www.readbyqxmd.com/read/29314736/development-of-a-chlamydia-suis-specific-antibody-enzyme-linked-immunosorbent-assay-based-on-the-use-of-a-b-cell-epitope-of-the-polymorphic-membrane-protein-c
#14
K De Puysseleyr, E Kieckens, L De Puysseleyr, H Van den Wyngaert, B Ahmed, S Van Lent, H H Creasy, G S A Myers, D Vanrompay
Chlamydia suis infections lead to economic loss in the pork industry. Chlamydia suis infections could be successfully treated with tetracyclines until the appearance of a tetracycline resistant phenotype, which was acquired via horizontal gene transfer of the tet(C) gene. Given the importance of C. suis as a swine pathogen and as a recently emerged tetracycline resistant pathogen with zoonotic potential, our aim was to develop a sensitive C. suis-specific antibody ELISA based on the polymorphic membrane proteins (Pmps)...
January 4, 2018: Transboundary and Emerging Diseases
https://www.readbyqxmd.com/read/29313467/temporal-expression-of-mutant-tdp-43-correlates-with-early-amyotrophic-lateral-sclerosis-phenotype-and-motor-weakness
#15
Qihua Chen, Jinxia Zhou, Cao Huang, Bo Huang, Fangfang Bi, Hongxia Zhou, Bo Xiao
BACKGROUND: Mutant transactive response DNA-binding protein (TDP-43) is closely correlated to inherited form of amyotrophic lateral sclerosis (ALS). TDP-43 transgenic rats can reproduce the core phenotype of ALS and constitutive expression of TDP-43 caused postnatal death. OBJECTIVE: To understand whether neurologic deficiency caused by mutant TDP-43 is dependent on its temporal expression. METHOD: Transgenic rats were established that express mutant human TDP-43 (M337V substitution) in neurons, then a Tet-off system was used to regulate its expression...
January 9, 2018: Current Neurovascular Research
https://www.readbyqxmd.com/read/29312823/a-historical-legacy-of-antibiotic-utilization-on-bacterial-seed-banks-in-sediments
#16
Laura Madueño, Christophe Paul, Thomas Junier, Zhanna Bayrychenko, Sevasti Filippidou, Karin Beck, Gilbert Greub, Helmut Bürgmann, Pilar Junier
The introduction of antibiotics for both medical and non-medical purposes has had a positive effect on human welfare and agricultural output in the past century. However, there is also an important ecological legacy regarding the use of antibiotics and the consequences of increased levels of these compounds in the environment as a consequence of their use and disposal. This legacy was investigated by quantifying two antibiotic resistance genes (ARG) conferring resistance to tetracycline (tet(W)) and sulfonamide (sul1) in bacterial seed bank DNA in sediments...
2018: PeerJ
https://www.readbyqxmd.com/read/29310046/chitosan-gel-sheet-containing-drug-carriers-with-controllable-drug-release-properties
#17
Tomoki Ito, Taku Takami, Yusuke Uchida, Yoshihiko Murakami
We prepared the "sheet-type hydrogel" (gel sheet), a sheet consisting of PEG-grafted chitosan and cross-linkable polymeric micelles, that were expected to be used for wound healing. We optimized the PEG-modification process, evaluated the strain-dependence of the gel's properties to obtain flexible gel sheets, and evaluated the drug-release properties of the gel sheets. Finally, we succeeded in observing that the release of the antibiotic tetracycline (TET) from the gel sheet in which TET existed only in the cross-linkers was lower than the release of TET from the sheet in which TET was dispersed in the polymer networks...
December 30, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/29307861/differences-in-tetracycline-resistance-determinants-carriage-among-shigella-flexneri-and-shigella-sonnei-are-not-related-to-different-plasmid-inc-type-carriage
#18
María J Pons, Alba Torrents de la Peña, Laura Mensa, Pilar Martón, Lidia Ruiz-Roldán, Sandra Martínez-Puchol, Jordi Vila, Joaquim Gascón, Joaquim Ruiz
OBJECTIVE: The aim of this study was to establish the different prevalence of the most common molecular mechanisms involved in tetracycline resistance as well as their relationship with plasmid incompatibility groups in a collection of Shigella spp. causing traveler's diarrhea. METHODS: Tetracycline susceptibility was established in 187 Shigella spp. (74S. flexneri and 113S. sonnei). Of these, 153 isolates were recovered as a confirmed cause of traveler's diarrhea...
January 4, 2018: Journal of Global Antimicrobial Resistance
https://www.readbyqxmd.com/read/29305959/enhanced-in-vitro-tigecycline-activity-in-the-presence-of-chelating-agents
#19
Amelia N Deitchman, Ravi Shankar Prasad Singh, Kenneth H Rand, Hartmut Derendorf
Lack of availability of novel antibiotic agents and the rise of resistance to existing therapies has led clinicians to utilize combination therapy to adequately treat bacterial infections. Here we examine how chelators may impact the in vitro activity of tigecycline (TIG) against Pseudomonas aeruginosa, Escherichia coli, and Klebsiella pneumoniae. Minimum inhibitory concentrations (MICs) were determined by broth dilution with and without various combinations of chelators (EDTA and other tetracyclines), and metal ions (i...
January 3, 2018: International Journal of Antimicrobial Agents
https://www.readbyqxmd.com/read/29296817/runx1-regulates-site-specificity-of-dna-demethylation-by-recruitment-of-dna-demethylation-machineries-in-hematopoietic-cells
#20
Takahiro Suzuki, Yuri Shimizu, Erina Furuhata, Shiori Maeda, Mami Kishima, Hajime Nishimura, Saaya Enomoto, Yoshihide Hayashizaki, Harukazu Suzuki
RUNX1 is an essential master transcription factor in hematopoietic development and plays important roles in immune functions. Although the gene regulatory mechanism of RUNX1 has been characterized extensively, the epigenetic role of RUNX1 remains unclear. Here, we demonstrate that RUNX1 contributes DNA demethylation in a binding site-directed manner in human hematopoietic cells. Overexpression analysis of RUNX1 showed the RUNX1-binding site-directed DNA demethylation. The RUNX1-mediated DNA demethylation was also observed in DNA replication-arrested cells, suggesting an involvement of active demethylation mechanism...
September 12, 2017: Blood Advances
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