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https://www.readbyqxmd.com/read/27914324/development-and-validation-of-species-specific-molecular-diagnostic-tool-for-opisthorchis-felineus-digenea-opisthorchiidae-metacercariae
#1
Monica Caffara, Laura Serracca, Andrea Gustinelli, Walter Vencia, Irene Rossini, Marino Prearo, Maria L Fioravanti
Opisthorchis felineus (family Opisthorchiidae) is a parasitic flatworm representing a serious threat to humans in some countries. Opisthorchiasis occurs after consumption of raw or undercooked cyprinid fish infected by the metacercarial stage of the parasite. Due to its small size, detection of the parasite in fish fillet is time-consuming and difficult. Furthermore, isolated metacercariae can be identified to genus but not to species level using morphological features and molecular techniques are necessary...
November 28, 2016: International Journal of Food Microbiology
https://www.readbyqxmd.com/read/27914209/methodological-flaws-introduce-strong-bias-into-molecular-analysis-of-microbial-populations
#2
Niclas Krakat, Reshma Anjum, Burak Demirel, Patrick Schröder
AIMS: Here we report how different cell disruption methods, PCR primers and in-silico analyses can seriously bias results from microbial population studies, with consequences for the credibility and reproducibility of the findings. Our results emphasize the pitfalls of commonly used experimental methods that can seriously weaken the interpretation of results. METHODS AND RESULTS: Four different cell lysis methods, three commonly used primer pairs and various computer-based analyses were applied to investigate the microbial diversity of a fermentation sample composed of chicken dung...
December 3, 2016: Journal of Applied Microbiology
https://www.readbyqxmd.com/read/27913973/emergence-of-new-virulent-rabbit-hemorrhagic-disease-virus-strains-in-saudi-arabia
#3
Mahmoud M Ismail, Mahmoud H A Mohamed, Ibrahim M El-Sabagh, Mohamed A Al-Hammadi
Rabbit hemorrhagic disease is an acute fatal highly contagious viral infectious disease that causes high losses among rabbitries. The disease was first reported in China in 1984 and later on in Saudi Arabia in 1996. The aim of this study was to investigate the emergence and pathogenicity of new rabbit hemorrhagic disease virus (RHDV) strains in Saudi Arabia. The pathogenicity was confirmed by inoculation in susceptible rabbits. Three RHDV strains were detected by reverse transcriptase polymerase chain reaction (RT-PCR) using primers targeting VP60 capsid protein gene in infected rabbitries during 2012 and 2013...
December 2, 2016: Tropical Animal Health and Production
https://www.readbyqxmd.com/read/27913826/partial-gene-sequencing-of-cyp1a-vitellogenin-and-metallothionein-in-mosquitofish-gambusia-yucatana-and-gambusia-sexradiata
#4
Roberto Vázquez-Euán, Karla S Escalante-Herrera, Gabriela Rodríguez-Fuentes
Ground characteristics in the Yucatan Peninsula make recovery and treatment of wastewater very expensive. This situation has contributed to an increase of pollutants in the aquifer. Unfortunately, studies related to the effects of those pollutants in native organisms are scarce. The aim of this work was to obtain partial sequences of widely known genes used as biomarkers of pollutant effect in Gambusia yucatana and Gambusia sexradiata. The studied genes were: cytochrome P450 1A (CYP1A); vitellogenin (VTG); metallothionein (MT), and two housekeeping genes, 18S and β-actin...
December 2, 2016: Bulletin of Environmental Contamination and Toxicology
https://www.readbyqxmd.com/read/27913413/new-arsenate-reductase-gene-arra-pcr-primers-for-diversity-assessment-and-quantification-in-environmental-samples
#5
Babur S Mirza, Darwin L Sorensen, R Ryan Dupont, Joan E McLean
: The extent of arsenic contamination in drinking water and its potential threat to human health have resulted in considerable research interest in the microbial species responsible for arsenic reduction. The arsenate reductase gene (arrA), an important component of the microbial arsenate reduction system, has been widely used as a biomarker to study arsenate reducing microorganisms. A new primer pair was designed and evaluated for qPCR and high-throughput sequencing of the arrA gene, because currently available PCR primers are not suitable for these applications...
December 2, 2016: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/27913276/association-of-genetic-polymorphisms-of-angiopoietin-like-4-with-severity-of-posttransplant-proteinuria-in-kidney-allograft-recipients
#6
Youngil Chang, Tariq Shah, Jaewook Yang, David I Min
BACKGROUND: Proteinuria is a hallmark of glomerular injury, and persistent proteinuria is associated with graft failure in kidney transplant patients. Recently, it is known that the level of circulating angiopoietin-like 4 (ANGPTL4) is elevated in the patients with human nephrotic syndrome, in which ANGPTL4 is responsible for relieving proteinuria. PURPOSE: The purpose of this study is to determine effects of clinical factors and genetic polymorphism of ANGPTL4 on proteinuria after kidney transplantation...
November 29, 2016: Transplant Immunology
https://www.readbyqxmd.com/read/27913256/rapid-upregulation-of-the-hippocampal-connexins-36-and-45-mrna-levels-during-memory-consolidation
#7
Siamak Beheshti, Reyhaneh Zeinali, Abolghasem Esmaeili
Gap junction channels are implicated in learning and memory process. However, their role on each of the particular stages of memory formation have been studied less. In this study, the time profile of the expression levels of hippocampal connexins 36 and 45 (Cx36 and Cx45) mRNAs was measured during memory consolidation, in a passive avoidance paradigm. Totally 36 adult male rats were distributed into 5 groups of each 6. At different times profiles (30min, 3, 6 and 24h) following training, rats were decapitated and their hippocampi were immediately removed and frozen in liquid nitrogen...
November 29, 2016: Behavioural Brain Research
https://www.readbyqxmd.com/read/27911895/dna-microarray-detection-of-18-important-human-blood-protozoan-species
#8
Mu-Xin Chen, Lin Ai, Jun-Hu Chen, Xin-Yu Feng, Shao-Hong Chen, Yu-Chun Cai, Yan Lu, Xiao-Nong Zhou, Jia-Xu Chen, Wei Hu
BACKGROUND: Accurate detection of blood protozoa from clinical samples is important for diagnosis, treatment and control of related diseases. In this preliminary study, a novel DNA microarray system was assessed for the detection of Plasmodium, Leishmania, Trypanosoma, Toxoplasma gondii and Babesia in humans, animals, and vectors, in comparison with microscopy and PCR data. Developing a rapid, simple, and convenient detection method for protozoan detection is an urgent need. METHODOLOGY/PRINCIPAL FINDINGS: The microarray assay simultaneously identified 18 species of common blood protozoa based on the differences in respective target genes...
December 2016: PLoS Neglected Tropical Diseases
https://www.readbyqxmd.com/read/27911492/rolling-circle-amplification-with-chemically-modified-nucleoside-triphosphates
#9
Marcel Hollenstein, Masad J Damha
Modified nucleoside triphosphates (dN*TPs) represent facile and versatile precursors for the introduction of chemical diversity into nucleic acids. While dN*TPs have been utilized in a plethora of practical applications, very little attention has been devoted to the assessment of their compatibility with isothermal amplification strategies. In this context, rolling circle amplification (RCA) is a wide-spread enzymatic replication method in which small single-stranded DNA (ssDNA) circles serve as templates in primer extension reactions yielding very long, ssDNA products...
December 1, 2016: Current Protocols in Nucleic Acid Chemistry
https://www.readbyqxmd.com/read/27910773/giant-oyster-mushroom-pleurotus-giganteus-agaricomycetes-enhances-adipocyte-differentiation-and-glucose-uptake-via-activation-of-ppar%C3%AE-and-glucose-transporters-1-and-4-in-3t3-l1-cells
#10
Puvaneswari Paravamsivam, Chua Kek Heng, Sri Nurestri Abdul Malek, Vikineswary Sabaratnam, Ravishankar Ram M, Umah Rani Kuppusamy
The edible mushroom Pleurotus giganteus was tested for its effect on adipocyte differentiation and glucose uptake activity in 3T3-L1 cells. The basidiocarps of P. giganteus were soaked in methanol to obtain a crude methanol extract and then fractionated to obtain an ethyl acetate extract. In this study, cell proliferation was measured using an MTT assay, lipid accumulation using an Oil Red O assay, and glucose uptake using a fluorescence glucose uptake assay. Gene expression was measured via real-time polymerase chain reaction analysis with TaqMan primer...
2016: International Journal of Medicinal Mushrooms
https://www.readbyqxmd.com/read/27910762/genetic-diversity-and-relationships-of-24-strains-of-genus-auricularia-agaricomycetes-assessed-using-srap-markers
#11
Ping Du, Tian-Xu Cao, Bao-Kai Cui, Yuan Yuan, Yu-Cheng Dai
Auricularia species are well known for their traditional edible and medicinal importance. In this study, sequence-related amplified polymorphism (SRAP) markers were used to evaluate the genetic diversity and relationships of 24 Auricularia strains obtained from different ecological regions of China. A total of 402 polymorphic loci were detected using 11 SRAP primer combinations with 100% polymorphic loci. The similarity coefficient ranged from 0.662 to 0.903, with an average of 0.73. Cluster analysis using the pair-group method using arithmetic average, based on the data from SRAP loci amplified by 11 primer pairs, showed that 24 Auricularia strains were distinguished into 4 groups with 73% similarity...
2016: International Journal of Medicinal Mushrooms
https://www.readbyqxmd.com/read/27908771/molecular-detection-and-genetic-characterization-of-babesia-theileria-and-anaplasma-amongst-apparently-healthy-sheep-and-goats-in-the-central-region-of-turkey
#12
Mo Zhou, Shinuo Cao, Ferda Sevinc, Mutlu Sevinc, Onur Ceylan, Sepil Ekici, Charoonluk Jirapattharasate, Paul Franck Adjou Moumouni, Mingming Liu, Guanbo Wang, Aiko Iguchi, Patrick Vudriko, Hiroshi Suzuki, Xuenan Xuan
Babesia spp., Theileria spp. and Anaplasma spp. are significant tick-borne pathogens of livestock globally. In this study, we investigated the presence and distribution of Babesia ovis, Theileria ovis and Anaplasma ovis in 343 small ruminants (249 sheep and 94 goats) from 13 towns in the Central Anatolia region of Turkey using species-specific PCR assays. The PCR were conducted using the primers based on the B. ovis ssu rRNA (BoSSUrRNA), T. ovis ssu rRNA (ToSSUrRNA) and A. ovis major surface protein 4 (AoMSP4) genes, respectively...
November 16, 2016: Ticks and Tick-borne Diseases
https://www.readbyqxmd.com/read/27908595/polymerase-chain-reaction-with-nearby-primers
#13
Ravil R Garafutdinov, Aizilya A Galimova, Assol R Sakhabutdinova
DNA analysis of biological specimens containing degraded nucleic acids such as mortal remains, archaeological artefacts, forensic samples etc. has gained more attention in recent years. DNA extracted from these samples is often inapplicable for conventional polymerase chain reaction (PCR), so for its amplification the nearby primers are commonly used. Here we report the data that clarify the features of PCR with nearby and abutting primers. We have shown that the proximity of primers leads to significant reduction of the reaction time and ensures the successful performance of DNA amplification even in the presence of PCR inhibitors...
November 28, 2016: Analytical Biochemistry
https://www.readbyqxmd.com/read/27908244/free-initiation-factors-eif4a-and-eif4b-are-dispensable-for-translation-initiation-on-uncapped-mrnas
#14
P A Sakharov, S Ch Agalarov
The formation of ribosomal 48S initiation complexes at the start AUG codon of uncapped mRNA leader sequences was studied using the methodology of primer extension inhibition (toe-printing). The experiments were performed in the system composed of purified individual components required for translation initiation. The formation of ribosomal 48S initiation complexes at the initiation codon was tested depending on the presence of the initiation factors eIF4F, eIF4A, and eIF4B. Several mRNAs containing short leader sequences lacking the extended secondary structure were studied...
October 2016: Biochemistry. Biokhimii︠a︡
https://www.readbyqxmd.com/read/27907856/cdna-aflp-analysis-of-transcripts-induced-in-chickpea-plants-by-tio2-nanoparticles-during-cold-stress
#15
Saeed Amini, Reza Maali-Amiri, Rahmat Mohammadi, Seyyedeh-Sanam Kazemi-Shahandashti
We evaluated the effect of TiO2 nanoparticles (NPs) on cold tolerance (CT) development in two chickpea (Cicer arietinum L.) genotypes (Sel96Th11439, cold tolerant, and ILC533, cold susceptible) by using cDNA-amplified fragment length polymorphism (cDNA-AFLP) technique during the first and sixth days of cold stress (CS) at 4 °C. Selective amplification by primer combinations generated 4200 transcript-derived fragments (TDFs) while 100 of them (2.62%) were differentially expressed. During CS, 60 differentially expressed TDFs of TiO2 NPs-treated plants were cloned and 10 of them produced successfully readable sequences...
November 22, 2016: Plant Physiology and Biochemistry: PPB
https://www.readbyqxmd.com/read/27907225/informativeness-of-ngs-analysis-for-vaginal-fluid-identification
#16
Saverio Giampaoli, Elisabetta DeVittori, Federica Valeriani, Andrea Berti, Vincenzo Romano Spica
The identification of vaginal fluids in forensic examinations plays an important role in crime scene reconstruction. Molecular detection of vaginal bacterial communities can lead to the correct discrimination of body fluids. These kinds of studies can be performed through multiplex real-time PCR using primers for a specific selection of bacteria. The availability of next-generation sequencing (NGS) protocols provided for the extension of the analysis to evaluate the prokaryotes present in specimens. In this study, DNA was extracted from 18 samples (vaginal, oral, fecal, yoghurt) and analyzed by real-time PCR and NGS...
December 1, 2016: Journal of Forensic Sciences
https://www.readbyqxmd.com/read/27907089/detection-of-human-papillomavirus-infection-in-patients-with-vaginal-intraepithelial-neoplasia
#17
Cristina Lamos, Charlotte Mihaljevic, Sebastian Aulmann, Thomas Bruckner, Christoph Domschke, Markus Wallwiener, Carmen Paringer, Herbert Fluhr, Sarah Schott, Christine Dinkic, Janina Brucker, Michael Golatta, Lisa Gensthaler, Michael Eichbaum, Christof Sohn, Joachim Rom
INTRODUCTION: Vaginal intraepithelial neoplasia (VAIN) is a pre-malignant lesion, potentially leading to vaginal cancer. It is a rare disease, representing less than 1% of all intraepithelial neoplasia of the female genital tract. Similar to cervical intraepithelial neoplasia (CIN), there are three different grades of VAIN. VAIN 1 is also known as a low-grade squamous intraepithelial lesion (LSIL), whereas VAIN 2 and VAIN 3 both represent high-grade squamous intraepithelial lesions (HSIL)...
2016: PloS One
https://www.readbyqxmd.com/read/27907026/asexual-recombinants-of-plasmopara-halstedii-pathotypes-from-dual-infection-of-sunflower
#18
Otmar Spring, Reinhard Zipper
Genetically homogenous strains of Plasmopara halstedii differing in host specificity and fungicide tolerance were used to test the hypothesis that asexual genetic recombination occurs and may account for the high genotype diversity of this homothallic reproducing oomycete, which causes downy mildew in sunflower. Dual inoculation of sunflower seedlings with single zoospore strains of complementary infection characteristics caused sporulation under conditions where inoculation with each strain alone failed to infect...
2016: PloS One
https://www.readbyqxmd.com/read/27907004/in-vitro-variant-surface-antigen-expression-in-plasmodium-falciparum-parasites-from-a-semi-immune-individual-is-not-correlated-with-var-gene-transcription
#19
Ellen Inga Bruske, Sandra Dimonte, Corinna Enderes, Serena Tschan, Matthias Flötenmeyer, Iris Koch, Jürgen Berger, Peter Kremsner, Matthias Frank
Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) is considered to be the main variant surface antigen (VSA) of Plasmodium falciparum and is mainly localized on electron-dense knobs in the membrane of the infected erythrocyte. Switches in PfEMP1 expression provide the basis for antigenic variation and are thought to be critical for parasite persistence during chronic infections. Recently, strain transcending anti-PfEMP1 immunity has been shown to develop early in life, challenging the role of PfEMP1 in antigenic variation during chronic infections...
2016: PloS One
https://www.readbyqxmd.com/read/27905281/identification-of-acanthamoeba-genotypes-in-pools-and-stagnant-water-in-ponds-in-sistan-region-in-southeast-iran
#20
Ali Aghajani, Mansour Dabirzadeh, Yahya Maroufi, Hossein Hooshyar
OBJECTIVE: Acanthamoeba is one of the most abundant free-living amoebas that is widely distributed in natural and artificial environment resources. Acanthamoeba pathogenic genotypes cause chronic human diseases including amoebic keratitis and granulomatous amoebic encephalitis. The aim of this study was to determine and identify Acanthamoeba genotypes residing in pools and stagnant water in ponds in Sistan region in southeast Iran. This descriptive study was conducted at the Parasitology Laboratory, School of Medicine, Zabol University of Medical Sciences...
September 2016: Türkiye Parazitolojii Dergisi
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