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Zaleha Shafiei, Zubaidah Haji Abdul Rahim, Koshy Philip, Nalina Thurairajah
BACKGROUND: Plant extracts mixture (PEM) and its individual constituent plant extracts(Psidium sp., Mangifera sp., Mentha sp.) are known to have an anti-adhering effect towards oral bacteria in the single-species biofilm. To date, the adhering ability of the early and late plaque colonisers (Streptococcus sanguinis and Streptococcus mutans) to PEM-treated experimental pellicle have not been investigated in dual-species biofilms. METHODS: Fresh leaves of these plants were used in the preparation of the respective aqueous extract decoctions...
2016: PeerJ
Yohei Kurosaki, Sayaka Okada, Sayuri Nakamae, Jiro Yasuda
Bovine papular stomatitis virus (BPSV) causes pustular cutaneous disease in cattle worldwide. This paper describes the development of a specific loop-mediated isothermal amplification (LAMP) assay to detect BPSV which did not cross-react with other parapoxviruses. To assess analytical sensitivity of this LAMP assay, DNA was extracted from serially diluted BPSV from which the infectious titer was determined by a novel assay based on calf kidney epithelial cells. The LAMP assay had equivalent analytical sensitivity to quantitative PCR, and could detect as few as 86 copies of viral DNA per reaction...
October 14, 2016: Journal of Virological Methods
Elena Parra, Ferran Segura, Jessica Tijero, Imma Pons, Maria-Mercedes Nogueras
A real-time PCR assay using SYBR Green was optimized to detect those Bartonella that are most frequently described as pathogens. The assay was genus-specific. Sequencing allowed to distinguish species. Assay sensitivity was determined using 10-fold serial dilutions of genomic DNA. Dynamic range was 100 ng-100 fg and sensitivity was 50 copies/reaction.
October 13, 2016: Molecular and Cellular Probes
Francesca Deidda, Angela Amoruso, Stefania Nicola, Teresa Graziano, Marco Pane, Serena Allesina, Elisabetta Raiteri, Mario Del Piano, Luca Mogna
GOALS: To investigate the possible use of Lactobacillus strains in the prophylaxis and/or adjuvant therapy of acute vulvovaginal candidiasis and other vaginal infections sustained by Candida yeasts. BACKGROUND: The incidence of Candida infections has substantially increased in recent years. Treatment of vaginal infections with lactobacilli has a long tradition, starting with Döderlein's description of the vaginal microbiota. MATERIALS AND METHODS: We assessed the activity of serially diluted fluconazole and miconazole (from 3 ng/mL to 1 mg/mL) against Candida strains...
November 2016: Journal of Clinical Gastroenterology
M A Landrum, N A Cox, D E Cosby, M E Berrang, S M Russell
New regulations and performance standards for Campylobacter have been implemented by the USDA - Food Safety and Inspection Service (FSIS). The objective of this study was to evaluate treatment with a low pH processing aid (CMS(1) PoultrypHresh), a formulated low pH processing aid, to reduce numbers of Campylobacter which could help companies meet regulatory requirements. Two experiments (3 replicates each) were conducted. Experiment 1, in each of 3 replicates, skin-on split chicken breasts (n = 15) were obtained from a local grocery and divided into groups of 5...
October 12, 2016: Poultry Science
Ludmila Juhásová, Ivica Králová-Hromadová, Eva Bazsalovicsová, Gabriel Minárik, Jan Štefka, Peter Mikulíček, Lenka Pálková, Margo Pybus
BACKGROUND: Fascioloides magna (Trematoda: Fasciolidae) is an important liver parasite of a wide range of free-living and domestic ruminants; it represents a remarkable species due to its large spatial distribution, invasive character, and potential to colonize new territories. The present study provides patterns of population genetic structure and admixture in F. magna across all enzootic regions in North America and natural foci in Europe, and infers migratory routes of the parasite on both continents...
October 13, 2016: Parasites & Vectors
Markus List, Marlene Pedersen Elnegaard, Steffen Schmidt, Helle Christiansen, Qihua Tan, Jan Mollenhauer, Jan Baumbach
High-throughput screening (HTS) has become an indispensable tool for the pharmaceutical industry and for biomedical research. A high degree of automation allows for experiments in the range of a few hundred up to several hundred thousand to be performed in close succession. The basis for such screens are molecular libraries, that is, microtiter plates with solubilized reagents such as siRNAs, shRNAs, miRNA inhibitors or mimics, and sgRNAs, or small compounds, that is, drugs. These reagents are typically condensed to provide enough material for covering several screens...
October 11, 2016: Journal of Biomolecular Screening
Stevan M Cokic, Peter Hoet, Lode Godderis, Martin Wiemann, Christof Asbach, Franz X Reichl, Jan De Munck, Bart Van Meerbeek, Kirsten L Van Landuyt
INTRODUCTION: Previous research revealed that during routine abrasive procedures like polishing, shaping or removing of composites, high amounts of respirable dust particles (<5μm) including nano-sized particles (<100nm) may be released. OBJECTIVE: To determine the cytotoxic potential of composite dust particles on bronchial epithelium cells. METHODS: Composite dust of five commercial composites (one nano-composite, two nano-hybrid and two hybrid composites) was generated following a clinically relevant protocol...
October 7, 2016: Dental Materials: Official Publication of the Academy of Dental Materials
Adriana Hellmeister de Campos Nogueira, Eliana De Stefano, Maira de Souza Nunes Martins, Liria Hiromi Okuda, Michele Dos Santos Lima, Thais da Silva Garcia, Otto Heinz Hellwig, José Eduardo Alves de Lima, Giovanni Savini, Edviges Maristela Pituco
Bluetongue (BT) is considered endemic in several regions of Brazil. The State of Sao Paulo was divided into 7 cattle production regions (circuits) according the different systems of breeding, operational and logistical capacity of the state veterinary service. At least 1 animal from each property (a total of 1,716 farms) was tested by competitive ELISA for the presence of antibodies against BTV. Sero‑positive sera were subsequently also tested by virus neutralization tests (VNT) using serial dilutions from 1:10 (cutoff) up to 1:640 (in MEM)...
September 30, 2016: Veterinaria Italiana
Jun Zhang, Sheng Yan, Dan Yuan, Qianbin Zhao, Say Hwa Tan, Nam-Trung Nguyen, Weihua Li
Separation of microparticles has found broad applications in biomedicine, industry and clinical diagnosis. In a conventional aqueous ferrofluid, separation of microparticles usually employs a sheath flow or two offset magnets to confine particle streams for downstream particle sorting. This complicates the fluid control, device fabrication, and dilutes the particle sample. In this work, we propose and develop a novel viscoelastic ferrofluid by replacing the Newtonian base medium of the conventional ferrofluid with non-Newtonian poly(ethylene oxide) (PEO) aqueous solution...
October 5, 2016: Lab on a Chip
Alicia M Haines, Shanan S Tobe, Adrian Linacre
Here, we evaluate Diamond Nucleic Acid Dye (DD) for use in quantitative PCR (qPCR) applications. Although DD is a commercially available stain for detection of DNA separated by gel electrophoresis, its use as a detection dye in qPCR has yet to be described. To determine if DD can be used in qPCR, we investigated its inhibitory effects on qPCR at concentrations ranging 0.1-2.5×. Serial dilution of DNA was used to determine the efficiency, sensitivity, and linearity of DD-generated qPCR data in comparison to other commonly used fluorescent dyes such as SYBR Green (SG), EvaGreen (EG), and BRYT Green (BG)...
October 1, 2016: BioTechniques
Gamou Fall, Martin Faye, Manfred Weidmann, Marco Kaiser, Anne Dupressoir, El Hadj Ndiaye, Yamar Ba, Mawlouth Diallo, Ousmane Faye, Amadou Alpha Sall
West Nile virus (WNV) is an emerging arbovirus, circulating worldwide between birds and mosquitoes, which impacts human and animal health. Since the mid-1990s, WNV outbreaks have emerged in Europe and America and represent currently the primary cause of encephalitis in the United States. WNV exhibits a great genetic diversity with at least eight different lineages circulating in the world, and four (1, 2, Koutango, and putative new) are present in Africa. These different WNV lineages are not readily differentiated by serology, and thus, rapid molecular tools are required for diagnostic...
October 6, 2016: Vector Borne and Zoonotic Diseases
Ao Huang, Xin Zhang, Shao-Lai Zhou, Ya Cao, Xiao-Wu Huang, Jia Fan, Xin-Rong Yang, Jian Zhou
PURPOSE: Circulating tumor DNA (ctDNA) is increasingly recognized as liquid biopsy to profile tumor genome. Droplet digital PCR (ddPCR) is a highly sensitive and easily operable platform for mutant detection. Here, we tried to detect ctDNA in hepatocellular carcinoma (HCC) patients using ddPCR. METHODS: Studies sequencing the genome of HCCs and COSMIC (Catalogue of Somatic Mutations in Cancer) database were reviewed to identify hotspot mutations. Circulating cell-free DNAs (cfDNAs) extracted from 1 ml preoperative plasma sample were analyzed to detect circulating mutants using ddPCR...
2016: Journal of Cancer
Lidwina Faraline Triprisila, Suharjono Suharjono, Antonius Christianto, Fatchiyah Fatchiyah
BACKGROUND: Goat milk is reported to have antimicrobial activity of several pathogen bacteria that contained on food materials. The research related with antimicrobial activity of Alpha-S2 casein from goat milk is relatively less than other casein components. Herein, we reported the antimicrobial activity of caprine Alpha-S2 Casein (CSN1S2) protein from Ethawah breed goat milk and yoghurt in Gram positive (Listeria monocytogenes, Staphylococcus aureus and Bacillus cereus) and negative pathogen bacteria (Escherichia coli, Salmonella typhi and Shigella flexneri)...
July 24, 2016: Materia Socio-medica
Hugh P Cam, Simon Whitehall
Reporter gene silencing assays provide a facile method for assessing the function of heterochromatin in Schizosaccharomyces pombe They use strains containing auxotrophic markers (commonly ura4(+) or ade6(+)) located within a heterochromatic region. Transcriptional silencing of these reporters can be assessed by plating serial dilutions of cells onto minimal agar. In addition, silencing of ura4(+) renders cells resistant to 5-fluoroorotic acid (5-FOA) and ade6(+) silencing results in red colony color on adenine-limiting agar...
October 3, 2016: Cold Spring Harbor Protocols
Tatsuro Iso, Takafumi Suzuki, Liam Baird, Masayuki Yamamoto
The transcription factor Nrf2 (NF-E2-related-factor-2) is essential for the oxidative and electrophilic stress responses. Keap1 (Kelch-like-ECH-associated-protein-1), an adaptor for a Cullin-3 (Cul3)-based ubiquitin ligase, regulates Nrf2 activity through proteasomal degradation, and acts as a sensor for oxidative and electrophilic stresses. The Keap1-Cul3 complex is a critical regulator of the cellular Nrf2 level, yet quantitative information regarding their endogenous intracellular concentrations in homeostatic conditions and during stress responses is unknown...
October 3, 2016: Molecular and Cellular Biology
Edward O Kwarteng, Krista M Heinonen
The gold standard definition of a hematopoietic stem cell (HSC) is a cell that when transferred into an irradiated recipient will have the ability to reestablish blood cell production for the lifespan of the recipient. This protocol explains how to set up a functional assay to compare the HSC capacities of two different populations of cells, such as bone marrow from mice of two different genotypes, and how to analyze the recipient mice by flow cytometry. The protocol uses HSC equivalents rather than cell sorting for standardization and discusses the advantages and disadvantages of both approaches...
2016: Journal of Visualized Experiments: JoVE
Jin Gao, Laura Couzens, Maryna C Eichelberger
Antibodies to neuraminidase (NA), the second most abundant surface protein on influenza virus, contribute toward protection against influenza. Traditional methods to measure NA inhibiting (NI) antibody titers are not practical for routine serology. This protocol describes the enzyme-linked lectin assay (ELLA), a practical alternative method to measure NI titers that is performed in 96 well plates coated with a large glycoprotein substrate, fetuin. NA cleaves terminal sialic acids from fetuin, exposing the penultimate sugar, galactose...
2016: Journal of Visualized Experiments: JoVE
Ali Baradaran Moghaddam, Masoud Alebouyeh, Nastaran Farzi, Saeed Bayati, Nour Amirmozafari
BACKGROUND: In this study, the efficacy of nitazoxanide in the treatment of Helicobacter pylori isolates, which were resistant to metronidazole, was examined. METHODS: One hundred twenty two patients who underwent endoscopy examinations at Kasra and Laleh hospitals in Tehran from November 2014 to July 2015 were enrolled. Helicobacter pylori strains were isolated from the patients' endoscopy biopsies by bacteriological culture. Those bacterial isolates resistant to metronidazole were examined for susceptibility to nitazoxanide...
2016: Medical Journal of the Islamic Republic of Iran
George D Calvey, Andrea M Katz, Chris B Schaffer, Lois Pollack
Knowledge of protein structure provides essential insight into function, enhancing our understanding of diseases and enabling new treatment development. X-ray crystallography has been used to solve the structures of more than 100 000 proteins; however, the vast majority represent long-lived states that do not capture the functional motions of these molecular machines. Reactions triggered by the addition of a ligand can be the most challenging to detect with crystallography because of the difficulty of synchronizing reactions to create detectable quantities of transient states...
September 2016: Structural Dynamics (Melville, N.Y.)
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