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https://www.readbyqxmd.com/read/28815494/use-of-the-dice-dual-integrase-cassette-exchange-system
#1
Alfonso P Farruggio, Mital S Bhakta, Michele P Calos
When constructing transgenic cell lines via plasmid DNA integration, precise targeting to a desired genomic location is advantageous. It is also often advantageous to remove the bacterial backbone, since bacterial elements can lead to the epigenetic silencing of neighboring DNA. The least cumbersome method to remove the plasmid backbone is recombinase-mediated cassette exchange (RMCE). RMCE is accomplished by arranging recombinase sites in the genome and in a donor plasmid such that a recombinase can both integrate the donor plasmid and excise its bacterial backbone...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28815492/building-cre-knockin-rat-lines-using-crispr-cas9
#2
Yuanwu Ma, Lianfeng Zhang, Xingxu Huang
Conditional gene inactivation strategy helps researchers to study the gene functions that are critical in embryogenesis or in defined tissues of adulthood. The Cre/loxP system is widely used for conditional gene inactivation/activation in cells or organisms. Cre knockin animal lines are essential for gene expression or inactivation in a spatially and temporally restricted manner. However, to generate a Cre knockin line by traditional approach is laborious. Recently, the clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9 (CRISPR/Cas9) has been proven as a simple and efficient genome-editing tool...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28812232/introgression-of-heat-shock-protein-hsp70-and-shsp-genes-into-the-malaysian-elite-chilli-variety-kulai-capsicum-annuum-l-through-the-application-of-marker-assisted-backcrossing-mab
#3
Magaji G Usman, Mohd Y Rafii, Mohammad Y Martini, Oladosu A Yusuff, Mohd R Ismail, Gous Miah
Backcrossing together with simple sequence repeat marker strategy was adopted to improve popular Malaysian chilli Kulai (Capsicum annuum L.) for heat tolerance. The use of molecular markers in backcross breeding and selection contributes significantly to overcoming the main drawbacks such as increase linkage drag and time consumption, in the ancient manual breeding approach (conventional), and speeds up the genome recovery of the recurrent parent. The strategy was adopted to introgress heat shock protein gene(s) from AVPP0702 (C...
August 15, 2017: Cell Stress & Chaperones
https://www.readbyqxmd.com/read/28811237/the-in-validity-of-volatile-poct-parameters-from-patients-beyond-normothermia
#4
Michael A Gruber, Stefan Felbermeir, Regina Lindner, Martin Kieninger
BACKGROUND: A common characteristic of all blood gas analyzers on the market is that measurements are processed at 37°C, not at the real patients´ temperature. Subsequently temperature-sensitive parameters can be mathematically corrected (alpha-stat method) or used directly (pH-stat method). National rules in Germany (Rili-BAEK) demand defined accuracy and precision without any restriction to samples´ temperatures or corrections. As consequence in the investigation at hand we tried to find out whether blood gas analyzers can fulfill the regulations for pCO2 and pO2 when normothermia of the matrix is not given...
August 12, 2017: Clinica Chimica Acta; International Journal of Clinical Chemistry
https://www.readbyqxmd.com/read/28810723/graft-survival-of-pediatric-kidney-transplant-recipients-selected-for-de-novo-steroid-avoidance-a-propensity-score-matched-study
#5
Edward J Nehus, Chunyan Liu, Bo Lu, Maurizio Macaluso, Mi-Ok Kim
Background: Steroid-avoidance protocols have gained popularity in pediatric kidney transplant recipients at low immunologic risk. The long-term safety of steroid avoidance in children with immunologic risk factors remains unknown. Methods: Pediatric kidney transplant recipients from 2004 to 2014 in the Organ Procurement and Transplantation Network database who received tacrolimus and mycophenolate immunosuppression were investigated. Propensity score matching was used to compare graft survival in 1624 children who received steroid avoidance with 1624 children who received steroid-based immunosuppression...
August 1, 2017: Nephrology, Dialysis, Transplantation
https://www.readbyqxmd.com/read/28804572/bacterial-infections-post-living-donor-liver-transplantation-in-egyptian-hepatitis-c-virus-cirrhotic-patients-a-single-center-study
#6
Mohamed F Montasser, Nadia A Abdelkader, Sara M Abdelhakam, Hany Dabbous, Iman F Montasser, Yasmine M Massoud, Waleed Abdelmoaty, Shereen A Saleh, Mohamed Bahaa, Hany Said, Mahmoud El-Meteini
AIM: To determine risk factors, causative organisms and antimicrobial resistance of bacterial infections following living-donor liver transplantation (LDLT) in cirrhotic patients. METHODS: This prospective study included 45 patients with hepatitis C virus-related end-stage liver disease who underwent LDLT at Ain Shams Center for Organ Transplant, Cairo, Egypt from January 2014 to November 2015. Patients were followed-up for the first 3 mo after LDLT for detection of bacterial infections...
July 18, 2017: World Journal of Hepatology
https://www.readbyqxmd.com/read/28801822/simultaneous-quantification-of-free-cholesterol-cholesteryl-esters-and-triglycerides-without-ester-hydrolysis-by-uhplc-separation-and-in-source-collision-induced-dissociation-coupled-ms-ms
#7
Michael S Gardner, Lisa G McWilliams, Jeffrey I Jones, Zsuzsanna Kuklenyik, James L Pirkle, John R Barr
We demonstrate the application of in-source nitrogen collision-induced dissociation (CID) that eliminates the need for ester hydrolysis before simultaneous analysis of esterified cholesterol (EC) and triglycerides (TG) along with free cholesterol (FC) from human serum, using normal phase liquid chromatography (LC) coupled to atmospheric pressure chemical ionization (APCI) tandem mass spectrometry (MS/MS). The analysis requires only 50 μL of 1:100 dilute serum with a high-throughput, precipitation/evaporation/extraction protocol in one pot...
August 11, 2017: Journal of the American Society for Mass Spectrometry
https://www.readbyqxmd.com/read/28795051/predictors-of-live-birth-and-pregnancy-success-after-in-vitro-fertilization-in-infertile-women-aged-40-and-over
#8
Hye Ok Kim, Nayoung Sung, In Ok Song
OBJECTIVE: The aim of this study was to evaluate pregnancy outcomes and the live birth rate at 1-year age increments in women aged ≥40 years undergoing fresh non-donor in vitro fertilization (IVF) and embryo transfer (ET), and to identify predictors of success in these patients. METHODS: This retrospective study was performed among women ≥40 years of age between 2004 and 2011. Of the 2,362 cycles that were conducted, ET was performed in 1,532 (73.1%). RESULTS: The clinical pregnancy rate and live birth rate in women ≥40 years significantly decreased with each year of increased age (p<0...
June 2017: Clinical and Experimental Reproductive Medicine
https://www.readbyqxmd.com/read/28790329/kdm4b-mediated-reduction-of-h3k9me3-and-h3k36me3-levels-improves-somatic-cell-reprogramming-into-pluripotency
#9
Jingwei Wei, Jisha Antony, Fanli Meng, Paul MacLean, Rebekah Rhind, Götz Laible, Björn Oback
Correct reprogramming of epigenetic marks is essential for somatic cells to regain pluripotency. Repressive histone (H) lysine (K) methylation marks are known to be stable and difficult to reprogram. In this study, we generated transgenic mice and mouse embryonic fibroblasts (MEFs) for the inducible expression of KDM4B, a demethylase that removes H3 K9 and H3K36 trimethylation (me3) marks (H3K9/36me3). Upon inducing Kdm4b, H3K9/36me3 levels significantly decreased compared to non-induced controls. Concurrently, H3K9me1 levels significantly increased, while H3K9me2 and H3K27me3 remained unchanged...
August 8, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28783901/chloramphenicol-interaction-with-functionalized-biochar-in-water-sorptive-mechanism-molecular-imprinting-effect-and-repeatable-application
#10
Mohammad Boshir Ahmed, John L Zhou, Huu Hao Ngo, Wenshan Guo, Md Abu Hasan Johir, Kireesan Sornalingam, M Sahedur Rahman
Biochar and functionalized biochar (fBC-1 and fBC-2) were prepared and applied to remove antibiotic chloramphenicol from deionized water, lake water and synthetic wastewater. Results showed that chloramphenicol removal on biochar was pH dependent and maximum sorption occurred at pH4.0-4.5. The sorption data of chloramphenicol fitted better with the Langmuir isotherm model than the Freundlich isotherm model with the maximum Langmuir sorption capacity of 233μMg(-1) using fBC-2. Chloramphenicol sorption on fBC-2 followed the trend: deionized water>lake water>synthetic wastewater...
July 31, 2017: Science of the Total Environment
https://www.readbyqxmd.com/read/28765581/oropharyngeal-mucosal-transmission-of-zika-virus-in-rhesus-macaques
#11
Christina M Newman, Dawn M Dudley, Matthew T Aliota, Andrea M Weiler, Gabrielle L Barry, Mariel S Mohns, Meghan E Breitbach, Laurel M Stewart, Connor R Buechler, Michael E Graham, Jennifer Post, Nancy Schultz-Darken, Eric Peterson, Wendy Newton, Emma L Mohr, Saverio Capuano, David H O'Connor, Thomas C Friedrich
Zika virus is present in urine, saliva, tears, and breast milk, but the transmission risk associated with these body fluids is currently unknown. Here we evaluate the risk of Zika virus transmission through mucosal contact in rhesus macaques. Application of high-dose Zika virus directly to the tonsils of three rhesus macaques results in detectable plasma viremia in all animals by 2 days post-exposure; virus replication kinetics are similar to those observed in animals infected subcutaneously. Three additional macaques inoculated subcutaneously with Zika virus served as saliva donors to assess the transmission risk from contact with oral secretions from an infected individual...
August 1, 2017: Nature Communications
https://www.readbyqxmd.com/read/28760670/standardization-of-8-color-flow-cytometry-across-different-flow-cytometer-instruments-a-feasibility-study-in-clinical-laboratories-in-switzerland
#12
Glier Hana, Heijnen Ingmar, Hauwel Mathieu, Dirks Jan, Quarroz Stéphane, Lehmann Thomas, Rovo Alicia, Arn Kornelius, Matthes Thomas, Hogan Cassandra, Keller Peter, Dudkiewicz Ewa, Stüssi Georg, Fernandez Paula
The EuroFlow Consortium developed a fully standardized flow cytometric approach from instrument settings, through antibody panel, reagents and sample preparation protocols, to data acquisition and analysis. The Swiss Cytometry Society (SCS) promoted a study to evaluate the feasibility of using such standardized measurements of 8-color data across two different flow cytometry platforms - Becton Dickinson (BD) FACSCanto II and Beckman Coulter (BC) Navios, aiming at increasing reproducibility and inter-laboratory comparability of immunophenotypic data in clinical laboratories in Switzerland...
July 28, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28759622/contamination-with-hiv-antibody-may-be-responsible-for-false-positive-results-in-specimens-tested-on-automated-platforms-running-hiv-4th-generation-assays-in-a-region-of-high-hiv-prevalence
#13
Diana Ruth Hardie, Stephen N Korsman, Nei-Yuan Hsiao, Molefi Daniel Morobadi, Sabeehah Vawda, Dominique Goedhals
INTRODUCTION: In South Africa where the prevalence of HIV infection is very high, 4th generation HIV antibody/p24 antigen combo immunoassays are the tests of choice for laboratory based screening. Testing is usually performed in clinical pathology laboratories on automated analysers. To investigate the cause of false positive results on 4th generation HIV testing platforms in public sector laboratories, the performance of two automated platforms was compared in a clinical pathology setting, firstly on routine diagnostic specimens and secondly on known sero-negative samples...
2017: PloS One
https://www.readbyqxmd.com/read/28758700/dengue-virus-infections-among-blood-donors-in-guangxi-of-china-2013-2014
#14
Z Gao, Y Zhang, Y Yang, M Xu, P Liao, W He, J Xu, Y Liu, M He
BACKGROUND: An unexpected, large dengue outbreak, spreading to 20 cities of Guangdong province, occurred in July 2014. Considering the overwhelming influence of this seasonal epidemic, whether this dengue virus outbreak has an significant impact on the blood donors of the ajacent areas in Guangdong requires at least a preliminary investigation. Liuzhou City of Guangxi was chosen as a good representative of the neighbouring geographical location of the dengue virus (DENV) outbreak areas in this research...
July 31, 2017: Transfusion Medicine
https://www.readbyqxmd.com/read/28756873/accordion-technique-combined-with-minimally-invasive-percutaneous-decortication-for-the-treatment-of-bone-non-union
#15
Qun Zhang, Wei Zhang, Zhuo Zhang, Peifu Tang, Lihai Zhang, Hua Chen
BACKGROUND: A variety of approaches have been used to treat oligotrophic or atrophic non-union. Conventional methods are often associated with great operative trauma, increased blood loss, a risk of re-infection, higher medical costs, and complications at the donor site. This study aimed to assess the clinical efficacy of the accordion technique combined with Minimally Invasive Percutaneous Decortication (MIPD) for these types of bone non-union. METHODS: From January 2010 to December 2015, 20 patients with long bone aseptic non-unions of the lower extremities without bone defects who were treated with the accordion technique combined with MIPD...
July 12, 2017: Injury
https://www.readbyqxmd.com/read/28751632/extracorporeal-membrane-oxygenation-support-for-rituximab-induced-acute-respiratory-distress-syndrome-in-an-abo-incompatible-living-donor-liver-transplant-recipient-successful-management-of-a-rare-complication-and-a-review-of-the-literature
#16
Ping-Chun Li, Ashok Thorat, Shih-Chao Hsu, Kin-Shing Poon, Horng-Ren Yang, Te-Hung Chen, Chun-Chieh Yeh, Long-Bin Jeng
BACKGROUND Rituximab is commonly used to reduce the agglutinin titer in ABO-incompatible liver transplant recipients. Although well-tolerated, rituximab infusion therapy may result in severe pulmonary adverse effects such as drug-induced pneumonitis, leading to acute respiratory distress syndrome (ARDS), which has a high mortality rate. Management of such rare cases in an ABO-incompatible patient has never been described before. Herein, we present successful use of extracorporeal membrane oxygenation (ECMO) support for rituximab-induced ARDS in an ABO-incompatible living donor liver transplantation (LDLT) recipient...
July 28, 2017: Annals of Transplantation: Quarterly of the Polish Transplantation Society
https://www.readbyqxmd.com/read/28746285/a-comparative-analysis-of-recombinant-human-bone-morphogenetic-protein-2-with-a-demineralized-bone-matrix-versus-iliac-crest-bone-graft-for-secondary-alveolar-bone-grafts-in-patients-with-cleft-lip-and-palate-review-of-501-cases
#17
Jeffrey A Hammoudeh, Artur Fahradyan, Daniel J Gould, Fan Liang, Thomas Imahiyerobo, Leo Urbinelli, JoAnna T Nguyen, William Magee, Stephen Yen, Mark M Urata
BACKGROUND: Alveolar cleft reconstruction using iliac crest bone graft is considered standard of care for children with complete cleft lip and palate at the time of mixed dentition. Harvesting bone may result in donor-site morbidity and additional operating time and length of hospitalization. Recombinant human bone morphogenetic protein (rhBMP)-2 with a demineralized bone matrix is an alternative bone source for alveolar cleft reconstruction. The authors investigated the outcomes of rhBMP-2/demineralized bone matrix versus iliac crest bone graft for alveolar cleft reconstruction by reviewing postoperative surgical complications and cleft closure...
August 2017: Plastic and Reconstructive Surgery
https://www.readbyqxmd.com/read/28743888/multiplication-rate-variation-in-the-human-malaria-parasite-plasmodium-falciparum
#18
Lee Murray, Lindsay B Stewart, Sarah J Tarr, Ambroise D Ahouidi, Mahamadou Diakite, Alfred Amambua-Ngwa, David J Conway
It is important to understand intrinsic variation in asexual blood stage multiplication rates of the most virulent human malaria parasite, Plasmodium falciparum. Here, multiplication rates of long-term laboratory adapted parasite clones and new clinical isolates were measured, using a newly standardised assay of growth from low starting density in replicate parallel cultures with erythrocytes from multiple different donors, across multiple cycles. Multiplication rates of long-term established clones were between 7...
July 25, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28734578/phosphatidylserine-is-the-signal-for-tam-receptors-and-their-ligands
#19
REVIEW
Greg Lemke
Nature repeatedly repurposes, in that molecules that serve as metabolites, energy depots, or polymer subunits are at the same time used to deliver signals within and between cells. The preeminent example of this repurposing is ATP, which functions as a building block for nucleic acids, an energy source for enzymatic reactions, a phosphate donor to regulate intracellular signaling, and a neurotransmitter to control the activity of neurons. A series of recent studies now consolidates the view that phosphatidylserine (PtdSer), a common phospholipid constituent of membrane bilayers, is similarly repurposed for use as a signal between cells and that the ligands and receptors of the Tyro3/Axl/Mer (TAM) family of receptor tyrosine kinases (RTKs) are prominent transducers of this signal...
July 19, 2017: Trends in Biochemical Sciences
https://www.readbyqxmd.com/read/28729350/structures-of-the-crispr-genome-integration-complex
#20
Addison V Wright, Jun-Jie Liu, Gavin J Knott, Kevin W Doxzen, Eva Nogales, Jennifer A Doudna
CRISPR-Cas systems depend on the Cas1-Cas2 integrase to capture and integrate short foreign DNA fragments into the CRISPR locus, enabling adaptation to new viruses. We present crystal structures of Cas1-Cas2 bound to both donor and target DNA in intermediate and product integration complexes, as well as a cryo-electron microscopy structure of the full CRISPR locus integration complex including the accessory protein Integration Host Factor (IHF). The structures show unexpectedly that indirect sequence recognition dictates integration site selection by favoring deformation of the repeat and the flanking sequences...
July 20, 2017: Science
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