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https://www.readbyqxmd.com/read/28437612/combinatorial-library-based-on-restriction-enzyme-mediated-modular-assembly
#1
Cuiping Ma, Chao Liang, Yifan Wang, Mei Pan, Qianqian Jiang, Chao Shi
Combinatorial approaches in directed evolution were proven to be more efficient for exploring sequence space and innovating the function of protein. We presented modular assembly of secondary structures (MASS) for constructing a combinatorial library. In this approach, secondary structure elements were extracted from natural existing proteins, the common linkers were flanking secondary structure elements, then digested by Hinf I restriction endonuclease that was used in the construction of combinatorial library for the first time...
April 24, 2017: ACS Combinatorial Science
https://www.readbyqxmd.com/read/28437139/protection-against-common-bean-rust-conferred-by-a-gene-silencing-method
#2
Bret Cooper, Kimberly B Campbell
Rust disease of the dry bean plant, Phaseolus vulgaris, is caused by the fungus Uromyces appendiculatus. The fungus acquires its nutrients and energy from bean leaves using a specialized cell structure, the haustorium, through which it secretes effector proteins that contribute to pathogenicity by defeating the plant immune system. Candidate effectors have been identified by DNA sequencing and motif analysis, and some candidates have been observed in infected leaves by mass spectrometry. To assess their roles in pathogenicity, we have inserted small fragments of genes for five candidates into bean pod mottle virus...
April 24, 2017: Phytopathology
https://www.readbyqxmd.com/read/28437106/allosteric-targeting-of-the-fanconi-anemia-ubiquitin-conjugating-enzyme-ube2t-by-fragment-screening
#3
Francesca E Morreale, Alessio Bortoluzzi, Viduth K Chaugule, Connor Arkinson, Helen Walden, Alessio Ciulli
Ube2T is the E2 ubiquitin-conjugating enzyme of the Fanconi anemia DNA repair pathway and it's overexpressed in several cancers, representing an attractive target for the development of inhibitors. Despite the extensive efforts in targeting the ubiquitin system, very few E2 binders have currently been discovered. Herein we report the identification of a new allosteric pocket on Ube2T through a fragment screening using biophysical methods. Several fragments binding to this site inhibit ubiquitin conjugation in vitro...
April 24, 2017: Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/28435821/utilization-of-pfge-as-a-powerful-discriminative-tool-for-the-investigation-of-genetic-diversity-among-mrsa-strains
#4
Solmaz Ohadian Moghadam, Mohammad Reza Pourmand, Masoumeh Douraghi, Samira Sabzi, Parisa Ghaffari
BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is a significant challenge to the burn patient. The implementation of proper monitoring programs and prompt identification of epidemic MRSA strains are critical to consequently control and eradicate potential outbreaks. This study aimed to define the genetic relatedness of MRSA strains isolated from burn patients by analyzing the large fragments of DNA. METHODS: In this cross-sectional study, 126 pus/wound swabs from skin and soft tissue infections (SSTIs) were collected from inpatients of Shahid Motahari Burn Center (Tehran, Iran) in 2013...
March 2017: Iranian Journal of Public Health
https://www.readbyqxmd.com/read/28435197/development-of-nested-polymerase-chain-reaction-based-diagnosis-of-duck-enteritis-virus-and-detection-of-dna-polymerase-gene-from-non-descriptive-duck-breeds-of-west-bengal-india
#5
Partha Sarathi Mandal, Sunit Kumar Mukhopadhayay, Saktipada Pradhan, Samiran Mondal, Chandrakanta Jana, Nimai Chandra Patra, Rabindra Nath Hansda
AIM: The study was undertaken to detect the clinical signs, postmortem lesions of embryonated duck plague (DP) infected eggs, and histopathological changes of chorioallantoic membrane (CAM) in non-descriptive ducks of West Bengal with special reference to standardize nested polymerase chain reaction (PCR). MATERIALS AND METHODS: After postmortem of suspected carcasses, samples were collected for virus isolation and identification through specific pathogen free (Khaki Campbell) embryonated duck eggs...
March 2017: Veterinary World
https://www.readbyqxmd.com/read/28434971/prevalence-and-genetic-characterization-of-toxoplasma-gondii-in-badgers-melogale-moschata-in-southern-china-by-pcr-rflp
#6
Kai Chen, Si-Yang Huang, Jin-Lei Wang, Rong-Liang Hu, Qiu-Xia Yao, Shou-Feng Zhang, Xing-Quan Zhu, Quan Liu
Toxoplasma gondii is an obligate intracellular apicomplexan parasite which is able to infect almost all warm-blooded animals. There is no information about the prevalence and genetic characterization of T. gondii in badgers (Melogale moschata) in China. Here, a total of 367 badgers were captured from different cities in Jiangxi province, Southern China. Genomic DNA was extracted from brain tissues of each badgers, and 57 (15.45%) of them were positive for T. gondii by semi-nested PCR of the B1 gene. The positive DNA samples were typed at 11 genetic markers, including 10 nuclear loci (SAG1, 5'-SAG2 and 3'-SAG2, alternative SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1) and an apicoplast locus (Apico), with multilocus polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technology...
April 20, 2017: Infection, Genetics and Evolution
https://www.readbyqxmd.com/read/28434050/prevalence-of-toxoplasma-gondii-infection-in-hiv-infected-patients-and-food-animals-and-direct-genotyping-of-t-gondii-isolates-southern-ghana
#7
Faustina Pappoe, Weisheng Cheng, Lin Wang, Yuanling Li, Dorcas Obiri-Yeboah, Samuel Victor Nuvor, Henock Ambachew, Xiaodong Hu, Qingli Luo, Deyong Chu, Yuanhong Xu, Jilong Shen
Toxoplasma gondii is of public health and veterinary importance causing severe diseases in immunocompromised individuals including HIV/AIDS patients and in congenital cases and animals. There is limited information on the epidemiology of T. gondii infection in humans, particularly HIV patients and food animals and the parasite genotypes in Ghana. A total of 394 HIV-infected patients from three hospitals were screened for T. gondii anti-IgG and IgM using ELISA. DNAs from blood samples of seropositve participants and 95 brain tissues of food animals were PCR assayed to detect Toxoplasma gra6...
April 22, 2017: Parasitology Research
https://www.readbyqxmd.com/read/28433807/silver-nanoparticle-induced-toxicity-to-human-sperm-by-increasing-ros-reactive-oxygen-species-production-and-dna-damage
#8
Enyin Wang, Yan Huang, Qingyun Du, Yingpu Sun
Silver nanoparticles (AgNPs) have been used in medical products and industrial coatings, due to their antimicrobial properties. Excessive use of AgNPs can have adverse effects on the human body, however, their toxicity characteristics to human sperm and the potential mechanisms are not entirely clear. In this study, we exposed human sperm to different doses of AgNPs (0, 50μgml(-1), 100μgml(-1), 200μgml(-1) or 400μgml(-1)) for various times (15min, 30min, or 60min), followed by analyses of the sperm viability, motility and the ratio of abnormal to normal sperm...
April 14, 2017: Environmental Toxicology and Pharmacology
https://www.readbyqxmd.com/read/28433754/mir-146a-suppresses-stat3-vegf-pathways-and-reduces-apoptosis-through-il-6-signaling-in-primary-human-retinal-microvascular-endothelial-cells-in-high-glucose-conditions
#9
Eun-Ah Ye, Jena J Steinle
microRNA (miRNA) play critical roles in the pathological processes of diabetic retinopathy, including inflammatory responses, insulin signaling, and angiogenesis. In addition to their regulatory functions on gene expression, miRNA is considered as a potential therapeutic target, as well as a diagnostic marker for many diseases. Our understanding on the pathological mechanisms underlying diabetic retinopathy is still incomplete and additional investigations are required to develop novel therapeutic strategies...
April 19, 2017: Vision Research
https://www.readbyqxmd.com/read/28433730/detection-of-rickettsia-helvetica-and-candidatus-r-tarasevichiae-dna-in-ixodes-persulcatus-ticks-collected-in-northeastern-european-russia-komi-republic
#10
Mikhail Yu Kartashov, Ludmila I Glushkova, Tamara P Mikryukova, Igor V Korabelnikov, Yulia I Egorova, Natalia L Tupota, Elena V Protopopova, Svetlana N Konovalova, Vladimir A Ternovoi, Valery B Loktev
The number of tick-borne infections in the northern European regions of Russia has increased considerably in the last years. In the present study, 676 unfed adult Ixodes persulcatus ticks were collected in the Komi Republic from 2011 to 2013 to study tick-borne rickettsioses. Rickettsia spp. DNA was detected by PCR in 51 (7.6%) ticks. The nucleotide sequence analysis of gltA fragments (765bp) from 51 ticks indicated that 60.8% and 39.2% of the ticks were infected with Rickettsia helvetica and Candidatus R. tarasevichiae, respectively...
April 6, 2017: Ticks and Tick-borne Diseases
https://www.readbyqxmd.com/read/28433723/elimination-of-the-cryptic-plasmid-in-leuconostoc-citreum-by-crispr-cas9-system
#11
Ye-Ji Jang, Seung-Oh Seo, Seul-Ah Kim, Ling Li, Tae-Jip Kim, Sun Chang Kim, Yong-Su Jin, Nam Soo Han
Leuconostoc spp. are important lactic acid bacteria for the fermentation of foods. In particular, L. citreum strains isolated from various foods have been used as host strains for genetic and metabolic engineering studies. In order to develop a food-grade genetic engineering system, L. citreum CB2567 was isolated from Kimchi. However, the isolated bacterium contained a cryptic plasmid which was difficult to eliminate. As the existence of the plasmid might hinder strain engineering, we eliminated the plasmid using an RNA-guided DNA endonuclease CRISPR/Cas9 system...
April 19, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28433512/effect-of-1-2-3-triazole-salts-non-classical-bioisosteres-of-miltefosine-on-leishmania-amazonensis
#12
Pedro H F Stroppa, Luciana M R Antinarelli, Arturene M L Carmo, Jacy Gameiro, Elaine S Coimbra, Adilson D da Silva
Here, we report the effect of new non-classical bioisosteres of miltefosine on Leishmania amazonensis. Fifteen compounds were synthesized and the compound dhmtAc, containing an acetate anion, a side chain of 10 carbon atoms linked to N-1 and a methyl group linked to N-3, showed high and selective biological activity against L. amazonensis. On the intracellular amastigotes, stages of the parasite related to human disease, the IC50 values were near or similar to the 1.0μM (0.9, 0.8 and 1.0μM on L. amazonensis-WT, and two transgenic L...
March 28, 2017: Bioorganic & Medicinal Chemistry
https://www.readbyqxmd.com/read/28433067/chloroquine-and-hydroxychloroquine-inhibit-bladder-cancer-cell-growth-by-targeting-basal-autophagy-and-enhancing-apoptosis
#13
Yi-Chia Lin, Ji-Fan Lin, Sheng-I Wen, Shan-Che Yang, Te-Fu Tsai, Hung-En Chen, Kuang-Yu Chou, Thomas I-Sheng Hwang
Chloroquine (CQ) and hydroxychloroquine (HCQ), two antimalarial drugs, are suggested to have potential anticancer properties. in the present study, we investigated the effects of CQ and HCQ on cell growth of bladder cancer with emphasis on autophagy inhibition and apoptosis induction in vitro. The results showed that CQ and HCQ inhibited the proliferation of multiple human bladder cell lines (including RT4, 5637, and T24) in a time- and dose-dependent fashion, especially in advanced bladder cancer cell lines (5637 and T24) compared to immortalized uroepithelial cells (SV-Huc-1) or other reference cancer cell lines (PC3 and MCF-7)...
May 2017: Kaohsiung Journal of Medical Sciences
https://www.readbyqxmd.com/read/28432551/partial-p53-dependence-of-anisomycin-induced-apoptosis-in-pc12-cells
#14
R Schipp, J Varga, J Bátor, M Vecsernyés, Z Árvai, M Pap, József Szeberényi
The bacterial antibiotic anisomycin is known to induce apoptosis by activating several mitogen-activated protein kinases and by inhibiting protein synthesis. In this study, the influence of p53 protein on the apoptosis-inducing effect of anisomycin was investigated. The effect of protein synthesis-inhibiting concentration of anisomycin on apoptotic events was analyzed using Western blot, DNA fragmentation, and cell viability assays in wild-type PC12 and in mutant p53 protein expressing p143p53PC12 cells. Anisomycin stimulated the main apoptotic pathways in both cell lines, but p143p53PC12 cells showed lower sensitivity to the drug than their wild-type counterparts...
April 21, 2017: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/28429848/stuck-in-fragments-population-genetics-of-the-endangered-collared-brown-lemur-eulemur-collaris-in-the-malagasy-littoral-forest
#15
Stefania Bertoncini, Jacopo D'Ercole, Francesca Brisighelli, Jean-Baptiste Ramanamanjato, Cristian Capelli, Sergio Tofanelli, Giuseppe Donati
OBJECTIVES: The Endangered collared brown lemur (Eulemur collaris) is the largest primate living in the littoral forest of southeastern Madagascar, a top priority habitat for biodiversity conservation on the island. Because this lemur is a key seed-disperser, an evaluation of the structure and connectivity of the populations surviving in the forest fragments is urgently needed to guide conservation plans. MATERIALS AND METHODS: Genetic variability at autosomal microsatellites and mitochondrial DNA was investigated in a total of 49 collared brown lemurs sampled by non-invasive methods in three littoral forest fragments and in the nearby lowland humid forest...
April 21, 2017: American Journal of Physical Anthropology
https://www.readbyqxmd.com/read/28429650/development-of-real-time-pcr-tests-for-the-detection-of-tenebrio-molitor-in-food-and-feed
#16
Frédéric Debode, Aline Marien, Amaury Gerard, Frédéric Francis, Olivier Fumiere, Gilbert Berben
Insects are rich in proteins and could be an alternative source of proteins to feed animals and humans. Numerous companies have started the production of insects for feed purposes. In Europe, these processed animal proteins are not yet authorized by legislation, as many questions still need to be answered concerning this "novel food". Authorizations will be possible when methods of authentication of the products are available. In this study we propose real-time PCR methods for the specific detection of the mealworm (Tenebrio molitor), one of the most widely used insects for food and feed production...
April 21, 2017: Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment
https://www.readbyqxmd.com/read/28429320/euglena-gracilis-genome-and-transcriptome-organelles-nuclear-genome-assembly-strategies-and-initial-features
#17
ThankGod Echezona Ebenezer, Mark Carrington, Michael Lebert, Steven Kelly, Mark C Field
Euglena gracilis is a major component of the aquatic ecosystem and together with closely related species, is ubiquitous worldwide. Euglenoids are an important group of protists, possessing a secondarily acquired plastid and are relatives to the Kinetoplastidae, which themselves have global impact as disease agents. To understand the biology of E. gracilis, as well as to provide further insight into the evolution and origins of the Kinetoplastidae, we embarked on sequencing the nuclear genome; the plastid and mitochondrial genomes are already in the public domain...
2017: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/28427878/localized-degradation-of-foreign-dna-strands-in-cells-only-excising-the-first-nucleotide-of-5-region
#18
Hui Li, Wei Shen, Michael Hon-Wah Lam, Haojun Liang
Intracellular delivery of foreign DNA probes sharply increases the efficiency of various biodetection protocols. Spherical nucleic acid (SNA) conjugate is a new type of probe that consists of a dense oligonucleotide shell attached typically to a gold nanoparticle core. They are widely used as novel labels for in vitro biodetection and intracellular assay. However, the degradation of foreign DNA still remains a challenge that can cause significant signal leakage (false positive signal). Hence, the site and behavior of intracellular degradation need to be investigated...
April 17, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/28426837/the-high-prevalence-of-hpv-and-hpv16-european-variants-in-cervical-and-anal-samples-of-hiv-seropositive-women-with-normal-pap-test-results
#19
Lays Paula Bondi Volpini, Neide Aparecida Tosato Boldrini, Luciana Bueno de Freitas, Angelica Espinosa Miranda, Liliana Cruz Spano
Human Immunodeficiency Virus (HIV)-seropositive women are more likely to have anogenital cancer, and high risk-HPV (HR-HPV) infection is the main associated factor. Between August 2013 and December 2015, we conducted a descriptive study to determine the HPV genotypes and HPV16 variants in cervical and anal samples of HIV-seropositive women with a normal Pap test. The viral DNA was amplified by PCR using the PGMY09/11 set of primers. Reverse line blot (RLB), restriction fragment length polymorphism (RFLP) and sequencing assays were used to determine the HPV genotypes...
2017: PloS One
https://www.readbyqxmd.com/read/28426159/open-microfluidic-gel-electrophoresis-rapid-and-low-cost-separation-and-analysis-of-dna-at-the-nanoliter-scale
#20
Ludwig Gutzweiler, Tobias Gleichmann, Laurent Tanguy, Peter Koltay, Roland Zengerle, Lutz Riegger
Gel electrophoresis is one of the most applied and standardized tools for separation and analysis of macromolecules and their fragments in academic research and in industry. In this work we present a novel approach for conducting on-demand electrophoretic separations of DNA molecules in open microfluidic (OM) systems on planar polymer substrates. The approach combines advantages of slab gel, capillary- & chip-based methods offering low consumable costs (<0.1 $) circumventing cost intensive microfluidic chip fabrication, short process times (5 minutes per analysis) and high sensitivity (4 ng/μl dsDNA) combined with reasonable resolution (17 bases)...
April 20, 2017: Electrophoresis
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