Post-training disruption of Arc protein expression in the anterior cingulate cortex impairs long-term memory for inhibitory avoidance training.
AuthorsHolloway CM, McIntyre CK,
JournalNeurobiol Learn Mem
PubMed ID21315825
'The activity-regulated-cytoskeletal-associated protein (Arc) has a well established role in memory consolidation and synaptic plasticity in the hippocampus and amygdala. However the role of Arc within the anterior cingulate cortex (ACC), an area of the brain involved in processing memory for pain, has yet to be examined. Here we sought ... More
Expression Analysis on Archival Material Revisited: Isolation and Quantification of RNA Extracted From FFPE Samples.
AuthorsDeben C, Zwaenepoel K, Boeckx C, Wouters A, Pauwels P, Peeters M, Lardon F, Baay M, Deschoolmeester V,
JournalDiagn Mol Pathol
PubMed ID23370428
'BACKGROUND:: Formalin-fixed paraffin-embedded (FFPE) tissue is the most readily available source of RNA for the gene expression studies. The main disadvantage is the poor quality of isolated RNA. Our group recently compared 5 commercially available RNA isolation kits and concluded that the RNeasy FFPE kit from Qiagen was the most ... More
A systematic evaluation of whole genome amplification of bisulfite-modified DNA.
'Studying DNA methylation profiles in detail should be the first step in epigenetic research. Although sodium bisulfite modification of genomic DNA is the gold standard method for DNA methylation analysis, this method results in the loss of the majority of the DNA material. Whole genome amplification (WGA) of bisulfite-modified DNA ... More
Investigation of the involvement of P-glycoprotein and multidrug resistance-associated protein 2 in the efflux of ximelagatran and its metabolites by using short hairpin RNA knockdown in Caco-2 cells.
AuthorsDarnell M, Karlsson JE, Owen A, Hidalgo IJ, Li J, Zhang W, Andersson TB,
JournalDrug Metab Dispos
PubMed ID20023051
'Liver and bile secretion can be an important first-pass and clearance route for drug compounds and also the site of several drug-drug interactions. In the clinical program for ximelagatran development, an unexpected effect of erythromycin on the pharmacokinetics of the direct thrombin inhibitor ximelagatran and its metabolites was detected. This ... More
[DNA extraction from coagulated human blood for application in genotyping techniques for human leukocyte antigen and immunoglobulin-like receptors].
'The objective of this study was to standardize a method for extracting high-quality DNA from samples of coagulated blood. Forty-eight samples of human coagulated blood were used for DNA extraction by means of the EZ-DNA commercial kit (Biological Industries, Beit Haemek, Israel), the Neoscience column kit (One Lambda Inc., San ... More
Potential role of multiple carbon fixation pathways during lipid accumulation in Phaeodactylum tricornutum.
'Phaeodactylum tricornutum is a unicellular diatom in the class Bacillariophyceae. The full genome has been sequenced (<30?Mb), and approximately 20 to 30% triacylglyceride (TAG) accumulation on a dry cell basis has been reported under different growth conditions. To elucidate P. tricornutum gene expression profiles during nutrient-deprivation and lipid-accumulation, cell cultures ... More
Comparative expression analysis of rpf-like genes of Mycobacterium tuberculosis H37Rv under different physiological stress and growth conditions.
AuthorsGupta RK, Srivastava BS, Srivastava R,
JournalMicrobiology
PubMed ID20522500
'Mycobacterium tuberculosis H37Rv possesses five resuscitation-promoting factors, RpfA-E, which are required for the resuscitation of dormancy in mycobacteria induced by prolonged incubation of the culture in stationary phase. This study explores the transcriptional profile of all the rpf-like genes of M. tuberculosis H37Rv in the exponential phase, stationary phase, non-culturable ... More
Prevalence of Toxoplasma gondii infection in stray and household cats in regions of Seoul, Korea.
AuthorsLee SE, Kim JY, Kim YA, Cho SH, Ahn HJ, Woo HM, Lee WJ, Nam HW,
JournalKorean J Parasitol
PubMed ID20877509
'The principal objective of this study was to investigate the prevalence of toxoplasmosis in household and stray cats in Seoul, Republic of Korea. We collected blood samples from 72 stray and 80 household cats, and all samples were examined by ELISA and nested PCR. The overall positive rates of Toxoplasma ... More
Detection of autotrophic verrucomicrobial methanotrophs in a geothermal environment using stable isotope probing.
AuthorsSharp CE, Stott MB, Dunfield PF,
JournalFront Microbiol
PubMed ID22912630
Genomic analysis of the methanotrophic verrucomicrobium
Mutations in EZH2 cause Weaver syndrome.
AuthorsGibson WT, Hood RL, Zhan SH, Bulman DE, Fejes AP, Moore R, Mungall AJ, Eydoux P, Babul-Hirji R, An J, Marra MA, Chitayat D, Boycott KM, Weaver DD, Jones SJ,
JournalAm J Hum Genet
PubMed ID22177091
We used trio-based whole-exome sequencing to analyze two families affected by Weaver syndrome, including one of the original families reported in 1974. Filtering of rare variants in the affected probands against the parental variants identified two different de novo mutations in the enhancer of zeste homolog 2 (EZH2). Sanger sequencing ... More
Global Proteomic Screening of Protein Allergens and Advanced Glycation Endproducts in Thermally Processed Peanuts.
AuthorsHebling CM, McFarland MA, Callahan JH, Ross MM,
JournalJ Agric Food Chem
PubMed ID23039025
Peanuts ( Arachis hypogaea ) are the cause of one of the most prevalent food allergies worldwide. Thermal processing (e.g., roasting) of peanuts and peanut-containing foods results in complex chemical reactions that alter structural conformations of peanut proteins, preventing accurate detection of allergens by most immunochemical and targeted screening methodologies. ... More
Bioreactor environment-sensitive sentinel genes as novel metrics for cell culture scale-down comparability.
AuthorsKondragunta B, Joshi BH, Han J, Brorson KA, Puri RK, Moreira AR, Rao G,
JournalBiotechnol Prog
PubMed ID22848039
Scale-down of bioreactors is currently done based on matching one or more measurable parameters such as k(L) a and P/V, which could result in insufficient process comparability. Currently, there is a lack of genomic translational studies in cell culture scale-down, which could help delineate measurable cellular attributes for improved scale-down. ... More
Differential neonatal testosterone imprinting of GH-dependent liver proteins and genes in female mice.
Abnormal exposure to steroid hormones within a critical developmental period elicits permanent alterations in female reproductive physiology in rodents, but the impact on the female GH axis and the underlying sexual differences in hepatic enzymes have not been described in detail. We have investigated the effect of neonatal androgenization of ... More
Simplifying and streamlining Escherichia coli-based cell-free protein synthesis.
AuthorsYang WC, Patel KG, Wong HE, Swartz JR,
JournalBiotechnol Prog
PubMed ID22275217
Escherichia coli cell-free protein synthesis (CFPS) uses E. coli extracts to make active proteins in vitro. The basic CFPS reaction mixture is comprised of four main reagent components: (1) energy source and CFPS chemicals, (2) DNA encoding the protein of interest, (3) T7 RNA Polymerase (RNAP) for transcription, and (4) ... More
Recombinant expression of two bacteriophage proteins that lyse clostridium perfringens and share identical sequences in the C-terminal cell wall binding domain of the molecules but are dissimilar in their N-terminal active domains.
AuthorsSimmons M, Donovan DM, Siragusa GR, Seal BS,
JournalJ Agric Food Chem
PubMed ID20825156
Clostridium perfringens is a Gram-positive anaerobic spore-forming bacterium capable of producing four major toxins that are responsible for disease symptoms and pathogenesis in a variety of animals, humans, and poultry. The organism is the third leading cause of human foodborne bacterial disease, and C. perfringens is the presumptive etiologic agent ... More
Automated extraction improves multiplex molecular detection of infection in septic patients.
AuthorsRegueiro BJ, Varela-Ledo E, Martinez-Lamas L, Rodriguez-Calviño J, Aguilera A, Santos A, Gomez-Tato A, Alvarez-Escudero J,
JournalPLoS One
PubMed ID20967222
Sepsis is one of the leading causes of morbidity and mortality in hospitalized patients worldwide. Molecular technologies for rapid detection of microorganisms in patients with sepsis have only recently become available. LightCycler SeptiFast test M(grade) (Roche Diagnostics GmbH) is a multiplex PCR analysis able to detect DNA of the 25 ... More
Development and validation of a SYBR-Green I real-time PCR protocol to detect hazelnut (Corylus avellana L.) in foods through calibration via plasmid reference standard.
AuthorsD'Andrea M, Coïsson JD, Travaglia F, Garino C, Arlorio M,
JournalJ Agric Food Chem
PubMed ID19891480
Many tree nuts are considered to be a serious problem in food safety, because of the presence of causative factors in IgE-mediated food allergies. Among these, hazelnut (Corylus avellana L.) seeds are largely used in a range of confectionery products and contain many well-characterized allergens. DNA-based methods and ELISA tests ... More
On-chip aptamer-based sandwich assay for thrombin detection employing magnetic beads and quantum dots.
AuthorsTennico YH, Hutanu D, Koesdjojo MT, Bartel CM, Remcho VT,
JournalAnal Chem
PubMed ID20545301
In this paper, we report the development of an on-chip aptamer-based fluorescence assay for protein detection and quantification based on sandwich ELISA principles. Thrombin was selected as a model analyte to validate the assay design, which involves two DNA thrombin aptamers recognizing two different epitopes of the protein. Aptamer-functionalized magnetic ... More
Memory-enhancing corticosterone treatment increases amygdala norepinephrine and Arc protein expression in hippocampal synaptic fractions.
Considerable evidence indicates that glucocorticoid hormones enhance the consolidation of memory for emotionally arousing events through interactions with the noradrenergic system of the basolateral complex of the amygdala (BLA). We previously reported that intra-BLA administration of a beta-adrenoceptor agonist immediately after inhibitory avoidance training enhanced memory consolidation and increased hippocampal ... More
The alternative oxidase (AOX) gene in Vibrio fischeri is controlled by NsrR and upregulated in response to nitric oxide.
AuthorsDunn AK, Karr EA, Wang Y, Batton AR, Ruby EG, Stabb EV,
JournalMol Microbiol
PubMed ID20487270
Alternative oxidase (AOX) is a respiratory oxidase found in certain eukaryotes and bacteria; however, its role in bacterial physiology is unclear. Exploiting the genetic tractability of the bacterium Vibrio fischeri, we explore the regulation of aox expression and AOX function. Using quantitative PCR and reporter assays, we demonstrate that aox ... More
Exposure of DNA and Bacillus subtilis spores to simulated martian environments: use of quantitative PCR (qPCR) to measure inactivation rates of DNA to function as a template molecule.
Several NASA and ESA missions are planned for the next decade to investigate the possibility of present or past life on Mars. Evidence of extraterrestrial life will likely rely on the detection of biomolecules, which highlights the importance of preventing forward contamination not only with viable microorganisms but also with ... More
Disruption of the dopamine d2 receptor impairs insulin secretion and causes glucose intolerance.
AuthorsGarcía-Tornadú I, Ornstein AM, Chamson-Reig A, Wheeler MB, Hill DJ, Arany E, Rubinstein M, Becu-Villalobos D,
JournalEndocrinology
PubMed ID20147524
The relationship between antidopaminergic drugs and glucose has not been extensively studied, even though chronic neuroleptic treatment causes hyperinsulinemia in normal subjects or is associated with diabetes in psychiatric patients. We sought to evaluate dopamine D2 receptor (D2R) participation in pancreatic function. Glucose homeostasis was studied in D2R knockout mice ... More
Differential expression of microRNAs between eutopic and ectopic endometrium in ovarian endometriosis.
AuthorsFiligheddu N, Gregnanin I, Porporato PE, Surico D, Perego B, Galli L, Patrignani C, Graziani A, Surico N,
JournalJ Biomed Biotechnol
PubMed ID20300586
Endometriosis, defined as the presence of endometrial tissue outside the uterus, is a common gynecological disease with poorly understood pathogenesis. MicroRNAs are members of a class of small noncoding RNA molecules that have a critical role in posttranscriptional regulation of gene expression by repression of target mRNAs translation. We assessed ... More
Concomitant administration of nitric oxide and glucocorticoids improves protection against bronchoconstriction in a murine model of asthma.
AuthorsJonasson S, Hedenstierna G, Hjoberg J,
JournalJ Appl Physiol
PubMed ID20538845
Glucocorticoids (GC) remain the first choice of treatment in asthma, but GC therapy is not always effective and is associated with side effects. In a porcine study in our laboratory, simultaneous administration of GC and nitric oxide (NO) attenuated the endotoxin-induced inflammatory response and made GC treatment more effective than ... More
Isolation of a novel lipase from a metagenomic library derived from mangrove sediment from the south Brazilian coast.
A novel gene coding for a LipA-like lipase with 283 amino acids and a molecular mass of 32 kDa was isolated and characterized from a metagenomic library prepared from mangrove sediment from the south Brazilian coast. LipA was 52% identical to a lipolytic enzyme from an uncultured bacterium and shared ... More
Targeted quantitation of overexpressed and endogenous cystic fibrosis transmembrane conductance regulator using multiple reaction monitoring tandem mass spectrometry and oxygen stable isotope dilution.
AuthorsJiang H, Ramos AA, Yao X,
JournalAnal Chem
PubMed ID19947594
Cystic fibrosis transmembrane conductance regulator (CFTR) functions as an ion channel in the apical plasma membrane of epithelial cells. Mutations in the gene coding for CFTR cause cystic fibrosis (CF). A major cellular dysfunction is insufficient apical plasma membrane expression of the protein. Its correction is important for developing new ... More
DNA qualification workflow for next generation sequencing of histopathological samples.
AuthorsSimbolo M, Gottardi M, Corbo V, Fassan M, Mafficini A, Malpeli G, Lawlor RT, Scarpa A,
Journal
PubMed ID23762227
Histopathological samples are a treasure-trove of DNA for clinical research. However, the quality of DNA can vary depending on the source or extraction method applied. Thus a standardized and cost-effective workflow for the qualification of DNA preparations is essential to guarantee interlaboratory reproducible results. The qualification process consists of the ... More