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https://www.readbyqxmd.com/read/28806770/increasing-creatine-kinase-activity-protects-against-hypoxia-reoxygenation-injury-but-not-against-anthracycline-toxicity-in-vitro
#1
Sevasti Zervou, Hannah J Whittington, Philip J Ostrowski, Fang Cao, Jack Tyler, Hannah A Lake, Stefan Neubauer, Craig A Lygate
The creatine kinase (CK) phosphagen system is fundamental to cellular energy homeostasis. Cardiomyocytes express three CK isoforms, namely the mitochondrial sarcomeric CKMT2 and the cytoplasmic CKM and CKB. We hypothesized that augmenting CK in vitro would preserve cell viability and function and sought to determine efficacy of the various isoforms. The open reading frame of each isoform was cloned into pcDNA3.1, followed by transfection and stable selection in human embryonic kidney cells (HEK293). CKMT2- CKM- and CKB-HEK293 cells had increased protein and total CK activity compared to non-transfected cells...
2017: PloS One
https://www.readbyqxmd.com/read/28804739/selection-of-genetically-modified-bacteriophages-using-the-crispr-cas-system
#2
Miriam Manor, Udi Qimron
We present a CRISPR-Cas based technique for deleting genes from the T7 bacteriophage genome. A DNA fragment encoding homologous arms to the target gene to be deleted is first cloned into a plasmid. The T7 phage is then propagated in Escherichia coli harboring this plasmid. During this propagation, some phage genomes undergo homologous recombination with the plasmid, thus deleting the targeted gene. To select for these genomes, the CRISPR-Cas system is used to cleave non-edited genomes, enabling isolation of the desired recombinant phages...
August 5, 2017: Bio-protocol
https://www.readbyqxmd.com/read/28803390/cancer-clonal-theory-immune-escape-and-their-evolving-roles-in-cancer-multi-agent-therapeutics
#3
REVIEW
Jonathan L Messerschmidt, Prianka Bhattacharya, Gerald L Messerschmidt
PURPOSE OF REVIEW: The knowledge base of malignant cell growth and resulting targets is rapidly increasing every day. Clonal theory is essential to understand the changes required for a cell to become malignant. These changes are then clues to therapeutic intervention strategies. Immune system optimization is a critical piece to find, recognize, and eliminate all cancer cells from the host. Only by administering (1) multiple therapies that counteract the cancer cell's mutational and externally induced survival traits and (2) by augmenting the immune system to combat immune suppression processes and by enhancing specific tumor trait recognition can cancer begin to be treated with a truly targeted focus...
August 12, 2017: Current Oncology Reports
https://www.readbyqxmd.com/read/28801903/synthetic-tumor-specific-promoters-for-transcriptional-regulation-of-viral-replication
#4
Maria Veronica Lopez, Eduardo G Cafferata, Diego L Viale, Osvaldo L Podhajcer
Here we describe a collection of methods that have been adapted to isolate and modify tumor-specific promoters (TSPs ) to drive viral replication for cancer therapy and other uses. We will describe as examples the secreted protein acidic and rich in cysteine (SPARC ) and the protease-activated receptor-1 (PAR-1) promoter. We outline strategies to select appropriate TSPs using bioinformatics resources and the methods utilized in their subsequent cloning, assessment of transcriptional activity, and their use in conditionally replicative oncolytic adenoviruses ...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28800944/systematic-review-tumor-associated-antigen-autoantibodies-and-ovarian-cancer-early-detection
#5
REVIEW
Renée Turzanski Fortner, Antje Damms-Machado, Rudolf Kaaks
OBJECTIVES: Tumor-associated autoantibodies (AAbs), produced as an immune response to tumor-associated antigens (TAAs), are a novel pathway of early detection markers. METHODS: We conducted a systematic review on AAbs and ovarian cancer to summarize the diagnostic performance of individual AAbs and AAb panels. A total of 29 studies including 85 AAbs were included; 27 of the studies were conducted in prevalent cases and cancer-free controls and 2 investigations included pre-diagnosis samples...
August 8, 2017: Gynecologic Oncology
https://www.readbyqxmd.com/read/28800723/a-clca-regulatory-protein-present-in-the-chemosensory-cilia-of-olfactory-sensory-neurons-induces-a-ca-2-activated-cl-current-when-transfected-into-hek293
#6
Casilda V Mura, Ricardo Delgado, María Graciela Delgado, Diego Restrepo, Juan Bacigalupo
BACKGROUND: CLCA is a family of metalloproteases that regulate Ca(2+)-activated Cl(-) fluxes in epithelial tissues. In HEK293 cells, CLCA1 promotes membrane expression of an endogenous Anoctamin 1 (ANO1, also termed TMEM16A)-dependent Ca(2+)-activated Cl(-) current. Motif architecture similarity with CLCA2, 3 and 4 suggested that they have similar functions. We previously detected the isoform CLCA4L in rat olfactory sensory neurons, where Anoctamin 2 is the principal chemotransduction Ca(2+)-activated Cl(-) channel...
August 11, 2017: BMC Neuroscience
https://www.readbyqxmd.com/read/28800630/measuring-single-constructs-by-single-items-constructing-an-even-shorter-version-of-the-short-five-personality-inventory
#7
Kenn Konstabel, Jan-Erik Lönnqvist, Sointu Leikas, Regina García Velázquez, Hiaying Qin, Markku Verkasalo, Gari Walkowitz
The aim of this study was to construct a short, 30-item personality questionnaire that would be, in terms of content and meaning of the scores, as comparable as possible with longer, well-established inventories such as NEO PI-R and its clones. To do this, we shortened the formerly constructed 60-item "Short Five" (S5) by half so that each subscale would be represented by a single item. We compared all possibilities of selecting 30 items (preserving balanced keying within each domain of the five-factor model) in terms of correlations with well-established scales, self-peer correlations, and clarity of meaning, and selected an optimal combination for each domain...
2017: PloS One
https://www.readbyqxmd.com/read/28800475/biological-immunological-and-functional-properties-of-two-novel-multi-variant-chimeric-recombinant-proteins-of-csp-antigens-for-vaccine-development-against-plasmodium-vivax-infection
#8
Samaneh H Shabani, Sedigheh Zakeri, Ali H Salmanian, Jafar Amani, Akram A Mehrizi, Georges Snounou, François Nosten, Chiara Andolina, Yousef Mourtazavi, Navid D Djadid
The circumsporozoite protein (CSP) of the malaria parasite Plasmodium vivax is a major pre-erythrocyte vaccine candidate. The protein has a central repeat region that belongs to one of repeat families (VK210, VK247, and the P. vivax-like). In the present study, computer modelling was employed to select chimeric proteins, comprising the conserved regions and different arrangements of the repeat elements (VK210 and VK247), whose structure is similar to that of the native counterparts. DNA encoding the selected chimeras (named CS127 and CS712) were synthetically constructed based on E...
August 8, 2017: Molecular Immunology
https://www.readbyqxmd.com/read/28800012/quality-assessment-of-prognostic-studies-using-cancer-stem-cell-markers-in-oral-squamous-cell-carcinoma
#9
Marina G Teixeira, Luciana Corrêa
Cancer stem cells (CSC) have been investigated as prognostic markers in oral squamous cell carcinoma (OSCC). However, an assessment of the reporting quality of these studies has not been performed yet. The aim of this study was to describe the reporting quality of prognostic studies involving CSCs and OSCC, focusing mainly on the immunohistochemical reproducibility. By means a systematic review, 34 articles were selected. Analyses of both general reporting quality and immunohistochemistry technique were performed by using checklists for multiple aspects related to study reproducibility...
August 9, 2017: Applied Immunohistochemistry & Molecular Morphology: AIMM
https://www.readbyqxmd.com/read/28799737/method-to-assemble-genomic-dna-fragments-or-genes-on-human-artificial-chromosome-with-regulated-kinetochore-using-a-multi-integrase-system
#10
Nicholas C O Lee, Jung-Hyun Kim, Nikolai Petrov, Hee-Sheung Lee, Hiroshi Masumoto, William C Earnshaw, Vladimir Larionov, Natalay Kouprina
The production of cells capable of carrying multiple transgenes to Mb-size genomic loci has multiple applications in biomedicine and biotechnology. In order to achieve this goal, three key steps are required: i) Cloning of large genomic segments; ii) Insertion of multiple DNA blocks at a precise location and iii) The capability to eliminate the assembled region from cells. In this study, we designed the Iterative Integration System (IIS) that utilizes recombinases Cre, ΦC31 and ΦBT1, and combined it with Human Artificial Chromosome (HAC) possessing a regulated kinetochore (alphoid(tetO)-HAC)...
August 11, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/28798470/identification-of-the-novel-3-utr-sequences-of-human-il-21-mrna-as-potential-targets-of-mirnas
#11
Yutaka Enomoto, Rie Takagi, Yutaka Naito, Tsuyoshi Kiniwa, Yasuhito Tanaka, Susumu Hamada-Tsutsumi, Masaaki Kawano, Sho Matsushita, Takahiro Ochiya, Atsushi Miyajima
Hepatitis B virus (HBV) infection is a leading cause of hepatocellular carcinoma worldwide. However, the strategy of HBV to escape from the host immune system remains largely unknown. In this study, we examined extracellular vesicles (EVs) secreted from human hepatocytes infected with HBV. EVs includeing exosomes are nano-size vesicles with proteins, mRNAs, and microRNAs (miRNAs), which can be transmitted to different cells. We found that 104 EV associated miRNAs were increased in hepatocytes more than 2-fold by HBV infection...
August 10, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28796894/subtractive-phage-display-selection-for-screening-and-identification-of-peptide-sequences-with-potential-use-in-serodiagnosis-of-paracoccidioidomycosis-caused-by-paracoccidioides-brasiliensis
#12
Leticia da Silva Portes, Erika Seki Kioshima, Zoilo Pires de Camargo, Wagner Luiz Batista, Patricia Xander
Paracoccidioidomycosis (PCM) is a systemic granulomatous disease endemic in Latin America whose aetiologic agents are the thermodimorphic fungi Paracoccidioides brasiliensis and Paracoccidioides lutzii. Despite technological advances, some problems have been reported for the fungal antigens used for serological diagnosis, and inconsistencies among laboratories have been reported. The use of synthetic peptides in the serological diagnosis of infectious diseases has proved to be a valuable strategy because in some cases, the reactions are more specific and sensitive...
August 10, 2017: Letters in Applied Microbiology
https://www.readbyqxmd.com/read/28795850/selection-and-characterization-of-antibody-clones-are-critical-for-accurate-flow-cytometry-based-monitoring-of-cd123-in-acute-myeloid-leukemia
#13
Nicole M Cruz, Mayumi Sugita, Nathan Ewing-Crystal, Linda Lam, Roman Galetto, Agnes Gouble, Julianne Smith, Duane C Hassane, Gail J Roboz, Monica L Guzman
No abstract text is available yet for this article.
August 10, 2017: Leukemia & Lymphoma
https://www.readbyqxmd.com/read/28794462/fear-of-predation-alters-clone-specific-performance-in-phloem-feeding-prey
#14
Mouhammad Shadi Khudr, Oksana Y Buzhdygan, Jana S Petermann, Susanne Wurst
Fear of predation has been shown to affect prey fitness and behaviour, however, to date little is known about the underlying genetics of responses to predator-associated risk. In an effort to fill this gap we exposed four naïve clones of green peach aphid (Myzus persicae), maintained on the model crop Brassica oleracea, to different types of cues from aphid lion (Chrysoperla carnea). The respective predation risks, we termed Fear Factors, were either lethal (consumption by predator), or non-lethal (non-consumptive predator-associated cues: plant-tethered predator cadavers and homogenised shoot-sprayed or soil-infused blends of predator remains)...
August 9, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28794126/a-refined-method-to-study-gene-dosage-changes-in-vitro-using-crispr-cas9
#15
Teresa P Raposo, Henry O Ebili, Mohammad Ilyas
AIMS: Gene dosage can have a major impact on cell biology, although, hitherto, it has been difficult to study using in vitro models. We sought to refine and accelerate the development of 'gene dosage' models through using CRISPR/Cas9 (a gene editing technology) for sequential knockout of gene alleles. METHODS: Our method involved (1) using Cas9 nuclease mRNA rather than expression plasmids, (2) using a fluorescently labelled FAM-6 tracr complexed with guide RNA and (3) using high-resolution melting (HRM) analysis to screen for mutations...
August 9, 2017: Journal of Clinical Pathology
https://www.readbyqxmd.com/read/28793859/investigating-knockdown-resistance-kdr-mechanism-against-pyrethroids-ddt-in-the-malaria-vector-anopheles-funestus-across-africa
#16
Helen Irving, Charles S Wondji
BACKGROUND: Understanding the molecular basis of insecticide resistance is key to improve the surveillance and monitoring of malaria vector populations under control. In the major malaria vector Anopheles funestus, little is currently known about the role of the knockdown resistance (kdr) mechanism. Here, we investigated the presence and contribution of knockdown resistance (kdr) to pyrethroids/DDT resistance observed in Anopheles funestus across Africa. RESULTS: Pyrosequencing genotyping and sequencing of the voltage gated sodium channel (VGSC) gene did not detect the common L1014F mutation in field collected An...
August 9, 2017: BMC Genetics
https://www.readbyqxmd.com/read/28793235/-r-r-butane-2-3-diol-dehydrogenase-from-bacillus-clausii-dsm-8716-t-cloning-and-expression-of-the-bdha-gene-and-initial-characterization-of-enzyme
#17
Lukas Muschallik, Denise Molinnus, Johannes Bongaerts, Martina Pohl, Torsten Wagner, Michael J Schöning, Petra Siegert, Thorsten Selmer
The gene encoding a putative (R,R)-butane-2,3-diol dehydrogenase (bdhA) from Bacillus clausii DSM 8716(T) was isolated, sequenced and expressed in Escherichia coli. The amino acid sequence of the encoded protein is only distantly related to previously studied enzymes (identity 33-43%) and exhibited some uncharted peculiarities. An N-terminally StrepII-tagged enzyme variant was purified and initially characterized. The isolated enzyme catalyzed the (R)-specific oxidation of (R,R)- and meso-butane-2,3-diol to (R)- and (S)-acetoin with specific activities of 12U/mg and 23U/mg, respectively...
August 6, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28791654/pharmacotherapy-of-myelofibrosis
#18
REVIEW
Douglas Tremblay, Bridget Marcellino, John Mascarenhas
Myelofibrosis (MF) is a myeloproliferative neoplasm that is pathologically characterized by bone marrow myeloproliferation, reticulin and collagen fibrosis, and extramedullary hematopoiesis. Constitutive activation of the Janus associated kinase (JAK)-signal transducers and activators of transcription signaling pathway with resultant elevation in pro-inflammatory cytokine levels is the pathogenic hallmark of MF. JAK inhibitors, namely ruxolitinib, have been successful in alleviating symptoms and reducing splenomegaly, but therapy-related myelosuppression has led to the further development of highly selective JAK2 inhibitors...
August 8, 2017: Drugs
https://www.readbyqxmd.com/read/28791368/two-novel-cross%C3%A2-protective-antigens-for-bovine-pasteurella-multocida
#19
Huihui Du, Chenlu Wu, Chunming Li, Rendong Fang, Jianwei Ma, Jiale Ji, Zhihong Li, Nengzhang Li, Yuanyi Peng, Zeyang Zhou
Pasteurella multocida is an important pathogen that leads to a range of diseases that have severe economic consequences on cattle production. In order to develop an effective cross‑protective component vaccine, an immunoproteomics approach was used to analyze outer membrane proteins (OMPs) of the P. multocida serotype A, B and F strains. Candidate antigen molecules from the whole genome were screened via linear trap quadrupole mass spectrometry and bioinformatics analysis, and the reactogenicity of the candidate antigen molecules was analyzed via cloning, expression, and ELISA or protein immunoblotting, and the vaccine efficacy of the candidate molecules was determined in infective animal models and cross‑protective antigens may be obtained...
August 3, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28790814/bacteriological-incidence-in-pneumonia-patients-with-pulmonary-emphysema-a-bacterial-floral-analysis-using-the-16s-ribosomal-rna-gene-in-bronchoalveolar-lavage-fluid
#20
Keisuke Naito, Kei Yamasaki, Kazuhiro Yatera, Kentaro Akata, Shingo Noguchi, Toshinori Kawanami, Kazumasa Fukuda, Takashi Kido, Hiroshi Ishimoto, Hiroshi Mukae
Pulmonary emphysema is an important radiological finding in chronic obstructive pulmonary disease patients, but bacteriological differences in pneumonia patients according to the severity of emphysematous changes have not been reported. Therefore, we evaluated the bacteriological incidence in the bronchoalveolar lavage fluid (BALF) of pneumonia patients using cultivation and a culture-independent molecular method. Japanese patients with community-acquired pneumonia (83) and healthcare-associated pneumonia (94) between April 2010 and February 2014 were evaluated...
2017: International Journal of Chronic Obstructive Pulmonary Disease
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