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https://www.readbyqxmd.com/read/29930713/critical-factors-for-lentivirus-mediated-prdx4-gene-transfer-in-the-hepg2-cell-line
#1
Afiah Nasuha Aznan, Norwahidah Abdul Karim, Wan Zurinah Wan Ngah, Zakiah Jubri
Optimization of critical factors affects transduction efficiency and is able to reduce reagent consumption. The present study aimed to determine the optimum transduction conditions of small hairpin (sh)RNA against peroxiredoxin 4 (PRDX4) in the HepG2 cell line. Cell viability assays were conducted based on serum condition, incubation time, polybrene concentration and antibiotic dose selection. Non-targeting control shRNA was transduced into HepG2 cells in a 5-fold serial dilution, and colonies positive for green fluorescent protein were counted using ImageJ software...
July 2018: Oncology Letters
https://www.readbyqxmd.com/read/29929824/a-novel-serological-assay-for-influenza-based-on-did-fluorescence-dequenching-that-is-free-from-observer-bias-and-potentially-automatable-a-proof-of-concept-study
#2
Alexander I Makarkov, Aakash R Patel, Valentine Bainov, Brian J Ward
BACKGROUND: Serum hemagglutination inhibition (HAI) and microneutralization (MN) antibodies are often used as a correlate of protection for influenza. However, these manual assays are labor-intensive and difficult to standardize due to variability in biologic reagents used and subjective interpretation of the results. METHODS: Sera with known HAI and MN titers were used to assess a novel test based on the inhibition of fluorescence 'dequenching'. Whole influenza virions (A/California/07/2009 (H1N1), A/Hong Kong/4801/2014 (H3N2) and B/Brisbane/60/2008) labelled with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindodicarbocyanine perchlorate (DiD) were exposed to serial dilutions of serum and mixed with turkey red blood cells followed by acidification of the media (pH 5...
June 18, 2018: Vaccine
https://www.readbyqxmd.com/read/29923972/rapid-detection-of-mycoplasma-pneumoniae-by-loop-mediated-isothermal-amplification-assay
#3
Xin Yuan, Changqing Bai, Qian Cui, Han Zhang, Jing Yuan, Kaiwen Niu, Yuzhong Feng, Xin Jin, Puyuan Li, Huiying Liu
To improve the diagnosis and treatment of Mycoplasma pneumoniae (Mp) infection and reduce the misuse of antibiotics, we sought to establish a loop-mediated isothermal amplification (LAMP) assay for rapid detection of Mp.Six primers specific for the Mp P1 gene were designed, and the LAMP method was used to rapidly detect Mp. The sensitivity of the LAMP method was determined by serial dilution of the standard Mp strain FH (standard strains of Mycoplasma pneumoniae). Specificity was assessed with 17 common pathogenic microorganisms in the respiratory tract...
June 2018: Medicine (Baltimore)
https://www.readbyqxmd.com/read/29920343/development-of-nested-multiplex-polymerase-chain-reaction-pcr-assay-for-the-detection-of-toxocara-canis-toxocara-cati-and-ascaris-suum-contamination-in-meat-and-organ-meats
#4
Zhenzhen Wang, Miho Shibata, Yen Thi Hoang Nguyen, Yayoi Hayata, Nariaki Nonaka, Haruhiko Maruyama, Ayako Yoshida
Ascarid Larva Migrans Syndrome (ascarid LMS) is a clinical syndrome in humans, caused by the migration of animal roundworm larvae such as Toxocara canis, Toxocara cati and Ascaris suum. Humans may acquire infection by ingesting embryonated eggs, or infective larvae of these parasites in contaminated meat and organ meats. To detect these pathogenic contaminations, a novel nested multiplex PCR system was developed. Our novel nested multiplex PCR assay showed specific amplification of T. canis, T. cati and Ascaris spp...
June 16, 2018: Parasitology International
https://www.readbyqxmd.com/read/29914957/extinction-of-zika-virus-and-usutu-virus-by-lethal-mutagenesis-reveals-different-patterns-of-sensitivity-to-three-mutagenic-drugs
#5
Maria Rosaria Bassi, Raquel Navarro Sempere, Prashansa Meyn, Charlotta Polacek, Armando Arias
Flaviviruses constitute an increasing source of public health concern with growing numbers of pathogens causing disease, and a geographic spread to temperate climates. Despite a large body of evidence supporting mutagenesis as a conceivable antiviral strategy, there is currently no data on the sensitivity to increased mutagenesis for Zika virus (ZIKV) and Usutu virus (USUV), two emerging flaviviral threats. In this study, we demonstrate that both viruses are sensitive to three ribonucleosides that have shown mutagenic activity against other RNA viruses - favipiravir, ribavirin and 5-fluorouracil - while they remain unaffected by a mutagenic deoxyribonucleoside...
June 18, 2018: Antimicrobial Agents and Chemotherapy
https://www.readbyqxmd.com/read/29904021/overcoming-resistance-of-human-non-hodgkin-s-lymphoma-to-cd19-car-ctl-therapy-by-celecoxib-and-histone-deacetylase-inhibitors
#6
Antoni Xavier Torres-Collado, Ali R Jazirehi
Patients with B-cell non-Hodgkin’s lymphoma (B-NHL) who fail to respond to first-line treatment regimens or develop resistance, exhibit poor prognosis. This signifies the need to develop alternative treatment strategies. CD19-chimeric antigen receptor (CAR) T cell-redirected immunotherapy is an attractive and novel option, which has shown encouraging outcomes in phase I clinical trials of relapsed/refractory NHL. However, the underlying mechanisms of, and approaches to overcome, acquired anti-CD19CAR CD8⁺ T cells (CTL)-resistance in NHL remain elusive...
June 14, 2018: Cancers
https://www.readbyqxmd.com/read/29899630/evaluation-of-the-mono-instrument-wave-one-mechanical-action-on-the-bacterial-load-reduction-in-vitro-study-of-32-permanent-human-teeth
#7
Anta Seck, Khaly Bane, Amadou Diop, Diouma Ndiaye, Seydina O Niang, Babacar Touré
Context: Enterococcus faecalis is the most common bacteria found in infected root canals. Aim: The purpose of this study was to evaluate the effect of the mono instrument (wave-one® ) on the reduction of E. faecalis in root canals. Materials and Methods: Thirty-two human monoroot teeth were used. After sterilization by autoclave, the teeth were infected by E. faecalis and incubated for 24 h. Each tooth underwent sampling before and after the root canal shaping...
May 2018: Journal of Conservative Dentistry: JCD
https://www.readbyqxmd.com/read/29888711/improved-sensitivity-and-limit-of-detection-using-a-receive-only-coil-in-magnetic-particle-imaging
#8
Hendrik Paysen, James Wells, Olaf Kosch, Uwe Steinhoff, Jochen Franke, Lutz Trahms, Tobias Schaeffter, Frank Wiekhorst
Magnetic Particle Imaging (MPI) is an imaging modality capable of quantitatively determining the 3D distribution of a magnetic nanoparticle (MNP) ensemble. In this work, we present a method for reducing the MNP limit of detection by employing a new receive-only coil (Rx-coil) for signal acquisition. The new signal detector is designed to improve the sensitivity and thus quality of reconstructed images. We present characterization measurements conducted with the prototype Rx-coil installed in a preclinical MPI scanner...
June 11, 2018: Physics in Medicine and Biology
https://www.readbyqxmd.com/read/29881423/-in-vitro-activity-of-trigonella-foenum-graecum-seeds-against-a-clinical-strain-of-acanthamoeba-genotype-t4
#9
Samira Dodangeh, Maryam Niyyati, Mohammad Kamalinejad, Jacob Lorenzo-Morales, Abdolali Moshfe, Ali Haghighi, Eznolah Azargashb
Acanthamoeba keratitis (AK) is a sight-threatening infection of the cornea disease that often presents with a lengthy and not fully effective treatment. Current therapeutic options against Acanthamoeba are not very effective against the cyst. Calibrated trophozoite/cyst suspension was incubated with the same volume of serial dilutions of the Trigonella foenum graecum aqueous extract (200, 250, 350, 450, 600, and 750 mg/mL) in microcentrifuge tubes and mixed by pipetting up and down. After that, the tubes were incubated at 26 ºC for 24, 48, and 72 h...
2018: Iranian Journal of Pharmaceutical Research: IJPR
https://www.readbyqxmd.com/read/29879421/high-dose-hook-effect-in-urinary-dcr2-assay-in-patients-with-chronic-kidney-disease
#10
Jia Chen, Ke-Hong Chen, Li-Ming Wang, Wei-Wei Zhang, Feng Lei, Huan-Zi Dai, Ya-Ni He
BACKGROUND: Urinary DcR2 (uDcR2) is a biomarker for the early detection the tubulointerstitial injury (TII) in patients with chronic kidney disease (CKD), but the high-dose hook effect may lead to falsely low or even negative results when using an enzyme-linked immunosorbent assay (ELISA). This study aimed to investigate if the high-dose hook effect exists with ELISA testing, and to uncover a potential approach for reducing this effect. METHODS: 72 CKD patients were recruited and categorized into four groups based on TII scores...
June 4, 2018: Clinical Biochemistry
https://www.readbyqxmd.com/read/29875567/antibacterial-efficacy-of-biosynthesized-silver-nanoparticles-against-enterococcus-faecalis-biofilm-an-in-vitro-study
#11
Kiran Rahul Halkai, Jayashree A Mudda, Vasundhara Shivanna, Vandana Rathod, Rahul Halkai
Aim: This study aims to evaluate the antibacterial efficacy of biosynthesized silver nanoparticles (AgNPs) produced using the fungi against Enterococcus faecalis biofilm model on root dentin. Materials and Methods: AgNPs were biosynthesized using the fungi Fusarium semitectum isolated from healthy leaves of Withania somnifera . Minimum inhibitory concentration (MIC) of AgNPs was determined by microbroth dilution method using series of dilutions. MIC dose was standardized to evaluate the antibacterial efficacy...
April 2018: Contemporary Clinical Dentistry
https://www.readbyqxmd.com/read/29869437/rhizospheric-bacterial-isolates-of-grass-pea-lathyrus-sativus-l-endowed-with-multiple-plant-growth-promoting-traits
#12
Adal Mussa, Tadege Million, Fassil Assefa
AIM: The main aim of this work was to examine the plant growth promoting, biocontrol activities and stress competency of grass pea rhizobacterial strains from Ethiopia. METHODS AND RESULTS: Serial dilutions were carried out to isolate rhizobacterial strains from the rhizosphere soil samples. The isolates were characterized for their plant growth promoting, biocontrolling and stress tolerance potential. The isolates that showed better performance in the tested parameters were identified by 16S rRNA gene sequencing...
June 5, 2018: Journal of Applied Microbiology
https://www.readbyqxmd.com/read/29859076/antibacterial-activity-and-in-situ-efficacy-of-bidens-pilosa-linn-and-dichrostachys-cinerea-wight-et-arn-extracts-against-common-diarrhoea-causing-waterborne-bacteria
#13
Pfarelo Daphney Shandukani, Shonisani Cathphonia Tshidino, Peter Masoko, Kgabo Maureen Moganedi
BACKGROUND: Bidens pilosa and Dichrostachys cinerea extracts were investigated for the antibacterial properties against waterborne diarrhoeagenic bacteria. METHODS: The plant materials were extracted using the direct and serial exhaustive methods using solvents of varying polarities, namely, hexane, dichloromethane, ethyl acetate, acetone and methanol. Qualitative phytochemical analysis and quantitative determination of total phenolic content of the leaf powders of the two plants were tested...
June 1, 2018: BMC Complementary and Alternative Medicine
https://www.readbyqxmd.com/read/29851964/development-of-a-micromanipulation-method-for-single-cell-isolation-of-prokaryotes-and-its-application-in-food-safety
#14
Marisa Hohnadel, Myriam Maumy, Renaud Chollet
For nearly a century, conventional microbiological methods have been standard practice for detecting and identifying pathogens in food. Nevertheless, the microbiological safety of food has improved and various rapid methods have been developed to overcome the limitations of conventional methods. Alternative methods are expected to detect low cell numbers, since the presence in food of even a single cell of a pathogenic organism may be infectious. With respect to low population levels, the performance of a detection method is assessed by producing serial dilutions of a pure bacterial suspension to inoculate representative food matrices with highly diluted bacterial cells (fewer than 10 CFU/ml)...
2018: PloS One
https://www.readbyqxmd.com/read/29845618/solvent-orange-60-is-a-potent-contact-sensitizer-in-occupational-and-everyday-life
#15
Kotryna Linauskienė, Erik Zimerson, Annarita Antelmi, Magnus Bruze, Lina Hagvall, Nils Hamnerius, Inese Hauksson, Kristina Ryberg, Marléne Isaksson
BACKGROUND: Solvent Orange (SO) 60 is a perinone-type dye that is often used in plastic materials such as spectacle frames and has been shown to cause contact allergy. The first case of SO 60 allergic contact dermatitis caused by spectacle frames was reported in 1999, and the second in 2011. We have recently seen 10 patients, of whom 6 developed dermatitis in the retroauricular/temporal area after wearing plastic spectacles. OBJECTIVES: To report the cause of the dermatitis in the 10 patients and to describe our first case with occupational SO 60 contact allergy...
May 30, 2018: Contact Dermatitis
https://www.readbyqxmd.com/read/29844722/pharmaceutical-significance-of-leuconostoc-mesenteroides-ks-tn11-isolated-from-nile-tilapia-oreochromis-niloticus
#16
Bilal Ahmad Paray, Irfan A Rather, Mohammad K Al-Sadoon, Al-Shammari Fanar Hamad
Aquatic animals are known for their myriad of beneficial bacteria with diverse biologically active compounds. The current study was aimed to isolate and characterize potentially beneficial lactic acid bacteria from Nile Tilapia and evaluate their pharmaceutical applications. The fish samples were dissected and stomach, intestine, and gills were collected and serially diluted for the isolation of lactic acid bacteria (LAB) on BCP agar media. Identification of isolate was carried by biochemical and molecular characterization using API kit and 16S rRNA gene sequencing analysis, respectively...
May 2018: Saudi Pharmaceutical Journal: SPJ: the Official Publication of the Saudi Pharmaceutical Society
https://www.readbyqxmd.com/read/29805209/standardization-and-application-of-real-time-polymerase-chain-reaction-for-rapid-detection-of-bluetongue-virus
#17
I Karthika Lakshmi, Kalyani Putty, Satya Samparna Raut, Sunil R Patil, P P Rao, B Bhagyalakshmi, Y Krishna Jyothi, B Susmitha, Y Vishnuvardhan Reddy, Sowmya Kasulanati, J Shiva Jyothi, Y N Reddy
Aim: The present study was designed to standardize real-time polymerase chain reaction (PCR) for detecting the bluetongue virus from blood samples of sheep collected during outbreaks of bluetongue disease in the year 2014 in Andhra Pradesh and Telangana states of India. Materials and Methods: A 10-fold serial dilution of Plasmid PUC59 with bluetongue virus (BTV) NS3 insert was used to plot the standard curve. BHK-21 and KC cells were used for in vitro propagation of virus BTV-9 at a TCID50/ml of 105 ml and RNA was isolated by the Trizol method...
April 2018: Veterinary World
https://www.readbyqxmd.com/read/29802191/experimental-evolution-of-escherichia-coli-k-12-at-high-ph-and-rpos-induction
#18
Issam Hamdallah, Nadia Torok, Katarina M Bischof, Nadim Majdalani, Sriya Chadalavada, Nonto Mdluli, Kaitlin E Creamer, Michelle Clark, Chase Holdener, Preston J Basting, Susan Gottesman, Joan L Slonczewski
Experimental evolution of Escherichia coli K-12 W3110 by serial dilutions for 2,200 generations at high pH extended the range of sustained growth from pH 9.0 to pH 9.3. pH 9.3-adapted isolates showed mutations in DNA-binding regulators and envelope proteins. One population showed an IS1 knockout of phoB (positive regulator of the phosphate regulon). A phoB :: kanR knockout increased growth at high pH. phoB mutants are known to increase production of fermentation acids, which could enhance fitness at high pH...
May 25, 2018: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/29793968/comparison-of-detection-limits-of-4-th-generation-combination-hiv-antigen-antibody-p24-antigen-and-viral-load-assays-on-diverse-hiv-isolates
#19
Mars Stone, John Bainbridge, Ana M Sanchez, Sheila M Keating, Andrea Pappas, Wes Rountree, Chris Todd, Sonia Bakkour, Mark Manak, Sheila A Peel, Robert W Coombs, Eric M Ramos, M Kathleen Shriver, Paul Contestable, Sangeetha Vijaysri Nair, David H Wilson, Martin Stengelin, Gary Murphy, Indira Hewlett, Thomas N Denny, Michael P Busch
BACKGROUND: Detection of acute HIV infection is critical for HIV public health and diagnostics. Clinical 4th generation antigen-antibody (Ag/Ab) combination (combo) and p24 Ag immunoassays have enhanced detection of acute infection compared to Ab alone assays, but require ongoing evaluation with currently circulating diverse subtypes. Genetically and geographically diverse HIV clinical isolates were used to assess clinical HIV diagnostic, blood screening and next generation assays. METHODS: Blinded 300 member panels of 20 serially diluted well-characterized antibody negative HIV isolates were distributed to manufacturers and end-user labs to assess relative analytic sensitivity of currently approved and pre-approved clinical HIV 4th generation Ag/Ab combo or p24 Ag alone immunoassays across diverse subtypes...
May 23, 2018: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/29792124/hypersensitivity-to-alemtuzumab-a-safe-and-effective-desensitization-protocol-a-case-report
#20
Diego Gutiérrez-Fernández, Mónica Saldaña-Valderas, Raquel de la Varga-Martínez, Antonio Foncubierta-Fernández, María J Fernández-Anguita, Maria Del C Fernández-Valle, Fermín Medina-Varo
We describe a successful desensitization to alemtuzumab in one patient diagnosed with T-cell prolymphocytic leukaemia. Alemtuzumab treatment was initiated during infusion number 18, the patient showed cutaneous eruption with a miliary pattern, despite premedication with corticosteroids and antihistamines. The eruption returned with successive alemtuzumab infusions (infusions 19, 20 and 21), remained present for longer and was more severe with each infusion. The patient was referred to our Allergy Unit as it was necessary to maintain alemtuzumab treatment...
January 1, 2018: Journal of Oncology Pharmacy Practice
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