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https://www.readbyqxmd.com/read/29147115/effects-of-aminoglycoside-antibiotics-on-human-embryonic-stem-cell-viability-during-differentiation-in-vitro
#1
Divya S Varghese, Shama Parween, Mustafa T Ardah, Bright Starling Emerald, Suraiya A Ansari
Human embryonic stem cells (hESCs) are being used extensively in array of studies to understand different mechanisms such as early human embryogenesis, drug toxicity testing, disease modeling, and cell replacement therapy. The protocols for the directed differentiation of hESCs towards specific cell types often require long-term cell cultures. To avoid bacterial contamination, these protocols include addition of antibiotics such as pen-strep and gentamicin. Although aminoglycosides, streptomycin, and gentamicin have been shown to cause cytotoxicity in various animal models, the effect of these antibiotics on hESCs is not clear...
2017: Stem Cells International
https://www.readbyqxmd.com/read/29143175/vectors-for-expression-of-signal-peptide-dependent-proteins-in-baculovirus-insect-cell-systems-and-their-application-to-expression-and-purification-of-the-high-affinity-immunoglobulin-gamma-fc-receptor-i-in-complex-with-its-gamma-chain
#2
Le T M Le, Jens R Nyengaard, Monika M Golas, Bjoern Sander
Integral membrane proteins play a central role in various cellular functions and are important therapeutic targets. However, technical challenges in the overexpression and purification of membrane proteins often represent a limiting factor for biochemical and structural studies. Here, we constructed a set of vectors, derivatives of MultiBac vectors that can be used to express proteins with a cleavable N-terminal signal peptide in insect cells. We propose these vectors for expression of type I membrane proteins and other secretory pathway proteins that require the signal recognition particle for translocation to the endoplasmic reticulum (ER)...
November 15, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/29120349/magnetic-beads-based-sensor-with-tailored-sensitivity-for-rapid-and-single-step-amperometric-determination-of-mirnas
#3
Eva Vargas, Rebeca M Torrente-Rodríguez, Víctor Ruiz-Valdepeñas Montiel, Eloy Povedano, María Pedrero, Juan J Montoya, Susana Campuzano, José M Pingarrón
This work describes a sensitive amperometric magneto-biosensor for single-step and rapid determination of microRNAs (miRNAs). The developed strategy involves the use of direct hybridization of the target miRNA (miRNA-21) with a specific biotinylated DNA probe immobilized on streptavidin-modified magnetic beads (MBs), and labeling of the resulting heteroduplexes with a specific DNA-RNA antibody and the bacterial protein A (ProtA) conjugated with an horseradish peroxidase (HRP) homopolymer (Poly-HRP40) as an enzymatic label for signal amplification...
November 9, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29112476/cross-sectional-evaluation-of-plasma-vitamin-d-levels-in-a-large-cohort-of-italian-patients-with-pediatric-autoimmune-neuropsychiatric-disorders-associated-with-streptococcal-infections
#4
Stefano Stagi, Gemma Lepri, Donato Rigante, Marco Matucci Cerinic, Fernanda Falcini
BACKGROUND: Pediatric autoimmune neuropsychiatric disorders associated with streptococcal infections (PANDAS) are immune-mediated diseases characterized by obsessive-compulsive symptoms and/or tics triggered by group A Streptococcus infections. Despite the well-known action of 25-hydroxyvitamin D [25(OH)D] on different conditions driven by systemic inflammation, there are no data about the 25(OH)D status in patients with PANDAS. AIMS: To evaluate plasma 25(OH)D levels in a large cohort of children and adolescents with PANDAS and comparing the results with healthy controls...
November 7, 2017: Journal of Child and Adolescent Psychopharmacology
https://www.readbyqxmd.com/read/29099353/streptococcus-bovimastitidis-sp-nov-isolated-from-a-dairy-cow-with-mastitis
#5
Stefan P W de Vries, Nazreen F Hadjirin, Elizabeth M Lay, Ruth N Zadoks, Sharon J Peacock, Julian Parkhill, Andrew J Grant, Scott McDougall, Mark A Holmes
Here we describe a new species of the genus Streptococcus that was isolated from a dairy cow with mastitis in New Zealand. Strain NZ1587(T) was Gram-positive, coccus-shaped and arranged as chains, catalase and coagulase negative, γ-haemolytic and negative for Lancefield carbohydrates (A-D, F and G). The 16S rRNA sequence did not match sequences in the NCBI 16S rRNA or GreenGenes databases. Taxonomic classification of strain NZ1587(T) was investigated using 16S rRNA and core genome phylogeny, genome-wide average nucleotide identity (ANI) and predicted DNA-DNA hybridisation (DDH) analyses...
November 3, 2017: International Journal of Systematic and Evolutionary Microbiology
https://www.readbyqxmd.com/read/29091937/phenome-wide-association-study-using-research-participants-self-reported-data-provides-insight-into-the-th17-and-il-17-pathway
#6
Margaret G Ehm, Jennifer L Aponte, Mathias N Chiano, Laura M Yerges-Armstrong, Toby Johnson, Jonathan N Barker, Suzanne F Cook, Akanksha Gupta, David A Hinds, Li Li, Matthew R Nelson, Michael A Simpson, Chao Tian, Linda C McCarthy, Deepak K Rajpal, Dawn M Waterworth
A phenome-wide association study of variants in genes in the Th17 and IL-17 pathway was performed using self-reported phenotypes and genetic data from 521,000 research participants of 23andMe. Results replicated known associations with similar effect sizes for autoimmune traits illustrating self-reported traits can be a surrogate for clinically assessed conditions. Novel associations controlling for a false discovery rate of 5% included the association of the variant encoding p.Ile684Ser in TYK2 with increased risk of tonsillectomy, strep throat occurrences and teen acne, the variant encoding p...
2017: PloS One
https://www.readbyqxmd.com/read/29069738/hematopoietic-transcription-factors-and-differential-cofactor-binding-regulate-prkacb-isoform-expression
#7
Olga N Kuvardina, Stefanie Herkt, Annekarin Meyer, Lucas Schneider, Jasmin Yillah, Nicole Kohrs, Halvard Bonig, Erhard Seifried, Carsten Müller-Tidow, Jörn Lausen
Hematopoietic differentiation is controlled by key transcription factors, which regulate stem cell functions and differentiation. TAL1 is a central transcription factor for hematopoietic stem cell development in the embryo and for gene regulation during erythroid/megakaryocytic differentiation. Knowledge of the target genes controlled by a given transcription factor is important to understand its contribution to normal development and disease. To uncover direct target genes of TAL1 we used high affinity streptavidin/biotin-based chromatin precipitation (Strep-CP) followed by Strep-CP on ChIP analysis using ChIP promoter arrays...
September 22, 2017: Oncotarget
https://www.readbyqxmd.com/read/29045629/signs-and-symptoms-of-group-a-versus-non-group-a-strep-throat-a-meta-analysis
#8
Thuy N Thai, Ariella P Dale, Mark H Ebell
Introduction: Both non-Group A streptococcal (non-GAS) pharyngitis and Group A streptococcal (GAS) pharyngitis are commonly found in patients with sore throat. It is not known whether or not they present with similar signs and symptoms compared to patients with non-streptococcal pharyngitis. Methods: MEDLINE was searched for prospective studies that reported throat culture for both GAS and non-GAS as a reference standard, and reported at least one sign, symptom, or the Centor score...
October 13, 2017: Family Practice
https://www.readbyqxmd.com/read/29022552/multiplex-pcr-for-identification-of-six-clinically-relevant-streptococci
#9
Rujirat Hatrongjit, Yukihiro Akeda, Shigeyuki Hamada, Marcelo Gottschalk, Anusak Kerdsin
PURPOSE: The aim of this study was to develop a multiplex PCR (mPCR) for simultaneous detection (single reaction) of six clinically relevant streptococcal species: Streptococcus pneumoniae, Streptococcus suis, Streptococcus gallolyticus subsp. gallolyticus, S. gallolyticus subsp. pasteurianus, Streptococcus intermedius and Streptococcus anginosus/constellatus. METHODS: mPCR with primers specific for S. pneumoniae (lytA), S. suis (recN), S. gallolyticus subsp. gallolyticus (tanB), S...
October 12, 2017: Journal of Medical Microbiology
https://www.readbyqxmd.com/read/29020531/mechanisms-of-antibiotic-resistance-in-bacteria-mediated-by-silver-nanoparticles
#10
Chitrada Kaweeteerawat, Preeyawis Na Ubol, Sanirat Sangmuang, Sasitorn Aueviriyavit, Rawiwan Maniratanachote
Silver nanoparticles (AgNPs) are widely used in industry, consumer products, and medical appliances due to their efficient antimicrobial properties. However, information on environmental toxicity and bacterial impact of these particles is not completely elucidated. Results showed that AgNPs produced growth inhibition and oxidative stress in bacteria Escherichia coli (gram negative) and Staphylococcus aureus (gram positive), with half-maximal inhibitory concentrations (IC50) of 12 and 7 mg/L, respectively. Surprisingly, bacteria pre-exposed to sublethal dose of AgNPs exhibited increased resistance toward antibiotics (ampicillin and Pen-Strep) with IC50 elevated by 3-13-fold...
October 11, 2017: Journal of Toxicology and Environmental Health. Part A
https://www.readbyqxmd.com/read/28971274/a-lactococcus-lactis-expression-vector-set-with-multiple-affinity-tags-to-facilitate-isolation-and-direct-labeling-of-heterologous-secreted-proteins
#11
Francisco Romero Pastrana, Jolanda Neef, Jan Maarten van Dijl, Girbe Buist
The gram-positive bacterium Lactococcus lactis is a useful host for extracellular protein production. A main advantage of L. lactis over other bacterial expression systems is that lactococcal cells display low levels of autolysis and proteolysis. Previously, we developed a set of vectors for nisin-inducible extracellular production of N- or C-terminally hexa-histidine (His6)-tagged proteins. The present study was aimed at expanding our portfolio of L. lactis expression vectors for protein purification and site-specific labeling...
November 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28966946/expression-and-purification-of-a-mammalian-p2x7-receptor-from-sf9-insect-cells
#12
Akira Karasawa, Toshimitsu Kawate
The P2X7 receptor is an extracellular ATP-gated ion channel found only in eukaryotes (Bartlett et al., 2014). Due to its unique properties among P2X receptors, such as formation of a large conductance pore, the P2X7 receptor has been implicated in devastating diseases like chronic pain (North and Jarvis, 2013). However, mechanisms underlying the P2X7 specific properties remain poorly understood, partly because purification of this eukaryotic membrane protein has been challenging. Here we describe a detailed protocol for expressing and purifying a mammalian P2X7 receptor using an insect cell-baculovirus system...
September 5, 2017: Bio-protocol
https://www.readbyqxmd.com/read/28964334/expression-and-purification-of-arrestin-in-yeast-saccharomyces-cerevisiae
#13
Ramona Schlesinger, Anneliese Cousin, Joachim Granzin, Renu Batra-Safferling
Protein purity and yield are two critical parameters for successful protein characterization using structural techniques such as X-ray crystallography, NMR, and several other biophysical methods. The yeast Saccharomyces cerevisiae is one of the popular eukaryotic model systems for overexpression and subsequent purification of recombinant proteins. Here, we describe a protocol for cloning, overexpression, purification, and crystallization of arrestin-1 and its splice variant p44 from yeast. The purification protocol involves a single-affinity chromatography step on a Strep-Tactin column...
2017: Methods in Cell Biology
https://www.readbyqxmd.com/read/28933822/rod-like-virus-based-multiarm-colloidal-molecules
#14
Alexis de la Cotte, Cheng Wu, Marie Trévisan, Andrii Repula, Eric Grelet
We report on the construction of multiarm colloidal molecules by tip-linking filamentous bacteriophages, functionalized either by biological engineering or chemical conjugation. The affinity for streptavidin of a genetically modified vector phage displaying Strep-tags fused to one end of the viral particle is measured by determining the dissociation constant, Kd. In order to improve both the colloidal stability and the efficiency of the self-assembly process, a biotinylation protocol having a chemical yield higher than 90% is presented to regioselectively functionalize the cystein residues located at one end of the bacteriophages...
October 24, 2017: ACS Nano
https://www.readbyqxmd.com/read/28932686/a-positive-rapid-strep-test-in-a-young-adult-with-acute-pharyngitis-be-careful-what-you-wish-for
#15
Burke A Cunha
In young adults with acute pharyngitis, the main differential diagnosis is between GAS pharyngitis, EBV infectious mononucleosis, or other causes of viral pharyngitis. A positive RST does not differentiate GAS colonization from infection as is well illustrated by this case. Laboratory test results must be interpreted in the appropriate clinical context to be diagnostically meaningful. The RST only detects group A streptococci, but does not, of itself, implicate a causative role in the patient's pharyngitis...
2017: IDCases
https://www.readbyqxmd.com/read/28928442/genome-wide-association-and-hla-region-fine-mapping-studies-identify-susceptibility-loci-for-multiple-common-infections
#16
Chao Tian, Bethann S Hromatka, Amy K Kiefer, Nicholas Eriksson, Suzanne M Noble, Joyce Y Tung, David A Hinds
Infectious diseases have a profound impact on our health and many studies suggest that host genetics play a major role in the pathogenesis of most of them. We perform 23 genome-wide association studies for common infections and infection-associated procedures, including chickenpox, shingles, cold sores, mononucleosis, mumps, hepatitis B, plantar warts, positive tuberculosis test results, strep throat, scarlet fever, pneumonia, bacterial meningitis, yeast infections, urinary tract infections, tonsillectomy, childhood ear infections, myringotomy, measles, hepatitis A, rheumatic fever, common colds, rubella and chronic sinus infection, in over 200,000 individuals of European ancestry...
September 19, 2017: Nature Communications
https://www.readbyqxmd.com/read/28923010/the-role-of-the-two-component-systems-cpx-and-arc-in-protein-alterations-upon-gentamicin-treatment-in-escherichia-coli
#17
Emina Ćudić, Kristin Surmann, Gianna Panasia, Elke Hammer, Sabine Hunke
BACKGROUND: The aminoglycoside antibiotic gentamicin was supposed to induce a crosstalk between the Cpx- and the Arc-two-component systems (TCS). Here, we investigated the physical interaction of the respective TCS components and compared the results with their respective gene expression and protein abundance. The findings were interpreted in relation to the global proteome profile upon gentamicin treatment. RESULTS: We observed specific interaction between CpxA and ArcA upon treatment with the aminoglycoside gentamicin using Membrane-Strep-tagged protein interaction experiments (mSPINE)...
September 18, 2017: BMC Microbiology
https://www.readbyqxmd.com/read/28918144/short-communication-molecular-epidemiology-of-streptococcus-agalactiae-differs-between-countries
#18
J Reyes, M Chaffer, Juan Carlos Rodriguez-Lecompte, Javier Sánchez, Ruth N Zadoks, Natasha Robinson, Ximena Cardona, N Ramírez, G P Keefe
Group B Streptococcus or Streptococcus agalactiae continue to be challenging for milk quality programs in countries with emerging dairy industries, such as Colombia, where high prevalence has been reported. Molecular typing of isolates is needed to understand the variability and epidemiology of this pathogen and to develop effective control and eradication programs. We characterized the molecular profile of Strep. agalactiae isolated from cows with subclinical mastitis in 21 Colombian dairy herds and measured diversity within and between herds using multilocus sequence typing...
November 2017: Journal of Dairy Science
https://www.readbyqxmd.com/read/28869231/how-a-mild-influenza-b-infection-can-kill-a-case-of-pulmonary-hemorrhage
#19
Nathalie Abi Hatem, Sabrina Campbell, Deepa Kuchelan, Jennifer Bowers, Edmundo Rubio
Viral influenza is a seasonal infection associated with significant morbidity and mortality. Rapidly fatal hemorrhagic pneumonia has been described in previously healthy individuals with β-hemolytic Streptococcus pneumoniae in a small series of patients, but it is not common in patients coinfected with influenza B and β-hemolytic Streptococcus, particularly since influenza B is considered less pathogenic than influenza A. However, despite being uncommon, this coinfection seems to be associated with high morbidity and mortality, particularly in healthy individuals...
September 2017: Lung India: Official Organ of Indian Chest Society
https://www.readbyqxmd.com/read/28859019/age-specific-antibiotic-prescribing-and-adherence-to-guidelines-in-pediatric-patients-in-primary-care
#20
Verica Ivanovska, Karin Hek, Aukje K Mantel-Teeuwisse, Hubert G M Leufkens, Liset van Dijk
BACKGROUND: Most antibiotics in children are used to treat viral and self-limiting conditions. This study aims to compare physicians' adherence to guidelines on antibiotic prescribing in fever and in ear and respiratory infections to children in different age groups in the Netherlands. METHODS: Data were used from the NIVEL Primary Care Database. For all pediatric episodes of fever, acute otitis media (AOM), streptococcal pharyngitis (strep throat), sinusitis, acute tonsillitis, acute bronchitis/bronchiolitis and pneumonia in 2012, we determined whether national guidelines were followed in regard to whether an antibiotic was prescribed, and the type of antibiotic...
August 28, 2017: Pediatric Infectious Disease Journal
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