Features of medullary thymic epithelium implicate postnatal development in maintaining epithelial heterogeneity and tissue-restricted antigen expression.
AuthorsGillard GO, Farr AG
JournalJ Immunol
PubMed ID16670287
'Although putative thymic epithelial progenitor cells have been identified, the developmental potential of these cells, the extent of medullary thymic epithelium (mTEC) heterogeneity, and the mechanisms that mediate the expression of a wide range of peripheral tissue-restricted Ags (TRAs) by mTECs remain poorly defined. Here we have defined several basic ... More
Diversity of human small intestinal Streptococcus and Veillonella populations.
Authorsvan den Bogert B, Erkus O, Boekhorst J, de Goffau M, Smid EJ, Zoetendal EG, Kleerebezem M
JournalFEMS Microbiol Ecol
PubMed ID23614882
'Molecular and cultivation approaches were employed to study the phylogenetic richness and temporal dynamics of Streptococcus and Veillonella populations in the small intestine. Microbial profiling of human small intestinal samples collected from four ileostomy subjects at four time points displayed abundant populations of Streptococcus spp. most affiliated with S. salivarius, S. thermophilus, ... More
Rediscovery of the Yesso scallop pathogen Perkinsus qugwadi in Canada, and development of PCR tests.
AuthorsItoh N, Meyer GR, Tabata A, Lowe G, Abbott CL, Johnson SC
Journal
PubMed ID23670082
'Perkinsus qugwadi, a pathogenic protozoan parasite of Yesso scallops Patinopecten yessoensis, is found only in cultured populations in British Columbia, Canada. This pathogen was first identified in 1988 and caused significant mortalities at some locations during the early 1990s. Prevalence of infection decreased dramatically following 1995, and the disease was ... More
Temporal role of Sertoli cell androgen receptor expression in spermatogenic development.
AuthorsHazra R, Corcoran L, Robson M, McTavish KJ, Upton D, Handelsman DJ, Allan CM
JournalMol Endocrinol
PubMed ID23160479
'Sertoli cell (SC) androgen receptor (AR) activity is vital for spermatogenesis. We created a unique gain-of-function transgenic (Tg) mouse model to determine the temporal role of SCAR expression in testicular development. The SC-specific rat Abpa promoter directed human Tg AR [Tg SC-specific AR (TgSCAR)] expression, providing strong premature postnatal AR ... More
P-glycoprotein in sheep liver and small intestine: gene expression and transport efflux activity.
AuthorsBallent M, Wilkens MR, Maté L, Muscher AS, Virkel G, Sallovitz J, Schröder B, Lanusse C, Lifschitz A
JournalJ Vet Pharmacol Ther
PubMed ID23409949
'The role of the transporter P-glycoprotein (P-gp) in the disposition kinetics of different drugs therapeutically used in veterinary medicine has been demonstrated. Considering the anatomo-physiological features of the ruminant species, the constitutive expression of P-gp (ABCB1) along the sheep gastrointestinal tract was studied. Additionally, the effect of repeated dexamethasone (DEX) ... More
A DNA methylation signature associated with the epigenetic repression of glycine N-methyltransferase in human hepatocellular carcinoma.
'The basic mechanisms underlying promoter DNA hypermethylation in cancer are still largely unknown. It has been proposed that the levels of the methyl donor group in DNA methylation reactions, S-adenosylmethionine (SAMe), might be involved. SAMe levels depend on the glycine-N-methyltransferase (GNMT), a one-carbon group methyltransferase, which catalyzes the conversion of ... More
Biotinylated photocleavable polyethylenimine: capture and triggered release of nucleic acids from solid supports.
AuthorsHandwerger RG, Diamond SL
JournalBioconjug Chem
PubMed ID17432825
'A biotinylated photocleavable polyethylenimine (B-PC-PEI) was designed and synthesized for the capture and controlled release of nucleic acids from solid supports. B-PC-PEI was synthesized via a three-step reaction process and verified by 1H NMR and mass spectrometry. In aqueous solution, the o-nitrobenzyl group within B-PC-PEI was efficiently cleaved by 5 ... More
Hyaluronan synthesis and myogenesis: a requirement for hyaluronan synthesis during myogenic differentiation independent of pericellular matrix formation.
'Exogenous hyaluronan is known to alter muscle precursor cell proliferation, migration, and differentiation, ultimately inhibiting myogenesis in vitro. The aim of the current study was to investigate the role of endogenous hyaluronan synthesis during myogenesis. In quantitative PCR studies, the genes responsible for synthesizing hyaluronan were found to be differentially ... More
Molecular characterization and expression analysis of chitinase from the Pacific oyster Crassostrea gigas.
AuthorsOkada Y, Yamaura K, Suzuki T, Itoh N, Osada M, Takahashi KG
JournalComp Biochem Physiol B Biochem Mol Biol
PubMed ID23507628
'Chitinases are necessary enzymes supporting functions as a host defense factor against chitin-coated pathogens. They also function as a digestive enzyme for the hydrolysis of dietary chitin. We conducted characterization and assessed the tissue expression of the encoding gene of a chitinase (EC 3.2.1.14), Cg-Chit1, and the production of recombinant ... More
Increased milk protein synthesis in response to exogenous growth hormone is associated with changes in mechanistic (mammalian) target of rapamycin (mTOR)C1-dependent and independent cell signaling.
AuthorsSciascia Q, Pacheco D, McCoard SA
JournalJ Dairy Sci
PubMed ID23462168
'The objective of this study was to determine if increased milk protein synthesis observed in lactating dairy cows treated with growth hormone (GH) was associated with mechanistic (or mammalian) target of rapamycin complex 1 (mTORC1) regulation of downstream factors controlling nucleocytoplasmic export and translation of mRNA. To address this objective, ... More
DNA shuffling of cytochrome P450 enzymes.
AuthorsBehrendorff JB, Johnston WA, Gillam EM
JournalMethods Mol Biol
PubMed ID23475678
'DNA family shuffling is an efficient method for creating libraries of novel enzymes, in which a high proportion of mutants exhibit correct folding and possess catalytic properties distinct from the starting material. The evolutionary arrangement of cytochromes P450 into subfamilies of enzymes with highly similar nucleotide sequences but distinct catalytic ... More
Genetic characterization of Theileria equi infecting horses in North America: evidence for a limited source of U.S. introductions.
AuthorsHall CM, Busch JD, Scoles GA, Palma-Cagle KA, Ueti MW, Kappmeyer LS, Wagner DM
JournalParasit Vectors
PubMed ID23399005
Theileria equi is a tick-borne apicomplexan hemoparasite that causes equine piroplasmosis. This parasite has a worldwide distribution but the United States was considered to be free of this disease until recently. ... More
Nucleospora cyclopteri n. sp., an intranuclear microsporidian infecting wild lumpfish, Cyclopterus lumpus L., in Icelandic waters.
AuthorsFreeman MA, Kasper JM, Kristmundsson Á
JournalParasit Vectors
PubMed ID23445616
Commercial fisheries of lumpfish Cyclopterus lumpus have been carried out in Iceland for centuries. Traditionally the most valuable part is the eggs which are harvested for use as a caviar substitute.Previously reported parasitic infections from lumpfish include an undescribed intranuclear microsporidian associated with abnormal kidneys and mortalities in captive lumpfish ... More
Underreporting of Dengue-4 in Brazil Due to Low Sensitivity of the NS1 Ag Test in Routine Control Programs.
We have identified fifty-eight samples that were positive for Dengue-4 among 119 samples with negative diagnoses for dengue via the Platelia™ dengue NS1 Ag in Aracaju, State of Sergipe, Brazil. We determined that the low sensitivity of the NS1 Ag test could be related to secondary dengue infections in the ... More
Development of a SYBR safe technique for the sensitive detection of DNA in cesium chloride density gradients for stable isotope probing assays.
AuthorsMartineau C, Whyte LG, Greer CW,
JournalJ Microbiol Methods
PubMed ID18329741
SYBR safe, a fluorescent nucleic acid stain, was evaluated as a replacement for ethidium bromide (EtBr) in cesium chloride (CsCl) density gradients for DNA stable isotope probing (DNA-SIP) assays. The separation of 12C- and 13C-labelled DNA using SYBR safe gave similar results to those obtained using EtBr with pure cultures ... More
Adaptation of yellow fever virus 17D to Vero cells is associated with mutations in structural and non-structural protein genes.
AuthorsBeasley DW, Morin M, Lamb AR, Hayman E, Watts DM, Lee CK, Trent DW, Monath TP
JournalVirus Res
PubMed ID23602827
Serial passaging of yellow fever virus 17D in Vero cells was employed to derive seed material for a novel inactivated vaccine, XRX-001. Two independent passaging series identified a novel lysine to arginine mutation at amino acid 160 of the envelope protein, a surface-exposed residue in structural domain I. A third ... More
Single step protocol to purify recombinant proteins with low endotoxin contents.
AuthorsReichelt P, Schwarz C, Donzeau M
JournalProtein Expr Purif
PubMed ID16290005
Endotoxin is an unwanted by product of recombinant proteins purified from Escherichia coli. The inherent toxicity of endotoxins makes their removal an important step for the proteins' application in several biological assays and for safe parenteral administration. The method described in this paper is a one-step protocol which is effective ... More
5-Methyltetrahydrofolate inhibits photosensitization reactions and strand breaks in DNA.
AuthorsOffer T, Ames BN, Bailey SW, Sabens EA, Nozawa M, Ayling JE
JournalFASEB J
PubMed ID17341682
The known functions of folate are to support one-carbon metabolism and to serve as photoreceptors for cryptochromes and photolyases. We demonstrate that 5-methyltetrahydrofolate (5-MTHF, the predominant folate in plasma) is also a potent, near diffusion limited, scavenger of singlet oxygen and quencher of excited photosensitizers. Both pathways result in decomposition ... More
Mechanisms of palatal epithelial seam disintegration by transforming growth factor (TGF) beta3.
AuthorsAhmed S, Liu CC, Nawshad A,
JournalDev Biol
PubMed ID17698055
TGFbeta3 signaling initiates and completes sequential phases of cellular differentiation that is required for complete disintegration of the palatal medial edge seam, that progresses between 14 and 17 embryonic days in the murine system, which is necessary in establishing confluence of the palatal stroma. Understanding the cellular mechanism of palatal ... More
The L-a-amino acid receptor GPRC6A is expressed in the islets of Langerhans but is not involved in L-arginine-induced insulin release.
AuthorsSmajilovic S, Clemmensen C, Johansen LD, Wellendorph P, Holst JJ, Thams PG, Ogo E, Bräuner-Osborne H
JournalAmino Acids
PubMed ID22714012
GPRC6A is a seven-transmembrane receptor activated by a wide range of L-a-amino acids, most potently by L-arginine and other basic amino acids. The receptor is broadly expressed, but its exact physiological role remains to be elucidated. It is well established that L-arginine stimulates insulin secretion; therefore, the receptor has been ... More
Development and evaluation of an IS711-based loop mediated isothermal amplification method (LAMP) for detection of Brucella spp. on clinical samples.
AuthorsPérez-Sancho M, García-Seco T, Arrogante L, García N, Martínez I, Diez-Guerrier A, Perales A, Goyache J, Domínguez L, Alvarez J
Journal
PubMed ID23714043
DNA-based methods have emerged as an additional tool for Brucella infection-confirmation at a herd level. However, their implementation may require the use of specialized equipment. In this context the recently developed loop-mediated isothermal amplification (LAMP) technique may constitute an additional and cost-effective tool for rapid and specific DNA detection, especially ... More
High-density yeast-tiling array reveals previously undiscovered introns and extensive regulation of meiotic splicing.
AuthorsJuneau K, Palm C, Miranda M, Davis RW
JournalProc Natl Acad Sci U S A
PubMed ID17244705
Knowing gene structure is vital to understanding gene function, and accurate genome annotation is essential for understanding cellular function. To this end, we have developed a genome-wide assay for mapping introns in Saccharomyces cerevisiae. Using high-density tiling arrays, we compared wild-type yeast to a mutant deficient for intron degradation. Our ... More
Denaturing gradient gel electrophoresis analysis of 16S ribosomal DNA to monitor changes in mouse gut bacterial communities during Salmonella enterica serovar Enteritidis latent infection.
AuthorsMaciel BM, Marques EL, Dias JC, Santos TF, Romano CC, Brendel M, Rezende RP
JournalGenet Mol Res
PubMed ID23546981
Changes in intestinal microbial flora during a 4-week period of Salmonella enterica serovar Enteritidis colonization in resistant mice (latent carrier animals) were evaluated using a culture independent method involving denaturing gradient gel electrophoresis. The contents of the ileocecal portion of the intestines produced 26 bands. Fifty-seven percent of the bands ... More
Combinatorial Domain Hunting: An effective approach for the identification of soluble protein domains adaptable to high-throughput applications.
AuthorsReich S, Puckey LH, Cheetham CL, Harris R, Ali AA, Bhattacharyya U, Maclagan K, Powell KA, Prodromou C, Pearl LH, Driscoll PC, Savva R
JournalProtein Sci
PubMed ID17008718
Exploitation of potential new targets for drug and vaccine development has an absolute requirement for multimilligram quantities of soluble protein. While recombinant expression of full-length proteins is frequently problematic, high-yield soluble expression of functional subconstructs is an effective alternative, so long as appropriate termini can be identified. Bioinformatics localizes domains, ... More
The persistence of biofilm-associated antibiotic resistance of Staphylococcus aureus isolated from clinical bovine mastitis cases in Australia.
AuthorsBabra C, Tiwari JG, Pier G, Thein TH, Sunagar R, Sundareshan S, Isloor S, Hegde NR, de Wet S, Deighton M, Gibson J, Costantino P, Wetherall J, Mukkur T
JournalFolia Microbiol (Praha)
PubMed ID23446489
The aim of this investigation was to determine the persistence of biofilm-associated antibiotic resistance developed by methicillin-sensitive Staphylococcus aureus (MSSA), of different capsular types, during biofilm formation. Because of superiority of the tissue culture plate (TCP) over the Congo Red Agar (CRA) method for measuring biofilm formation, it was used ... More
A mutant alphaII-spectrin designed to resist calpain and caspase cleavage questions the functional importance of this process in vivo.
AuthorsMeary F, Metral S, Ferreira C, Eladari D, Colin Y, Lecomte MC, Nicolas G
JournalJ Biol Chem
PubMed ID17374614
alpha- and beta-spectrins are components of molecular scaffolds located under the lipid bilayer and named membrane skeletons. Disruption of these scaffolds through mutations in spectrins demonstrated that they are involved in the membrane localization or the maintenance of proteins associated with them. The ubiquitous alphaII-spectrin chain bears in its central ... More
Vascular endothelial growth factors signalling in normal human dental pulp: a study of gene and protein expression.
AuthorsVirtej A, Løes S, Iden O, Bletsa A, Berggreen E
JournalEur J Oral Sci
PubMed ID23489898
In the well-vascularized dental pulp vascular endothelial growth factor A (VEGF-A) is expressed. Vascular endothelial growth factor A is a member of the VEGF family, which includes VEGFs-B, -C, and -D. The latter three have not been investigated in the pulp. Vascular endothelial growth factors C and D are the ... More
Switch from caspase-dependent to caspase-independent death during heart development: essential role of endonuclease G in ischemia-induced DNA processing of differentiated cardiomyocytes.
AuthorsBahi N, Zhang J, Llovera M, Ballester M, Comella JX, Sanchis D
JournalJ Biol Chem
PubMed ID16754658
Differentiated cardiomyocytes are resistant to caspase-dependent cell death; however, the mechanisms involved are still uncertain. We previously reported that low Apaf1 expression partially accounts for cardiomyocyte resistance to apoptosis. Here, we extend the knowledge on the molecular basis of cardiac resistance to caspase activation by showing that the whole caspase-dependent ... More
AFLP-based transcript profiling (cDNA-AFLP) for genome-wide expression analysis.
AuthorsVuylsteke M, Peleman JD, van Eijk MJ
JournalNat Protoc
PubMed ID17545977
Although DNA microarrays are currently the standard tool for genome-wide expression analysis, their application is limited to organisms for which the complete genome sequence or large collections of known transcript sequences are available. Here, we describe a protocol for cDNA-AFLP, an AFLP-based transcript profiling method that allows genome-wide expression analysis ... More
Successful transmission of Solenopsis invicta virus 3 to Solenopsis invicta fire ant colonies in oil, sugar, and cricket bait formulations.
AuthorsValles SM, Porter SD, Choi MY, Oi DH
JournalJ Invertebr Pathol
PubMed ID23602901
Tests were conducted to evaluate whether Solenopsis invicta virus 3 (SINV-3) could be delivered in various bait formulations to fire ant colonies and measure the corresponding colony health changes associated with virus infection in Solenopsis invicta. Three bait formulations (10% sugar solution, cricket paste, and soybean oil adsorbed to defatted ... More
Use of denaturing gradient gel electrophoresis for the identification of mixed oral yeasts in human saliva.
AuthorsWeerasekera MM, Sissons CH, Wong L, Anderson S, Holmes AR, Cannon RD
JournalJ Med Microbiol
PubMed ID23065546
A PCR-denaturing gradient gel electrophoresis (DGGE) method was established for the simultaneous presumptive identification of multiple yeast species commonly present in the oral cavity. Published primer sets targeting different regions of the Saccharomyces cerevisiae 26-28S rRNA gene (denoted primer sets N and U) and the 18S rRNA gene (primer set ... More
Functional Expression of Purinergic P2 Receptors and Transient Receptor Potential Channels by Human Urothelium.
Further to its role as a physical barrier, the urothelium is considered to play an active role in mechanosensation. A key mechanism is the release of transient mediators that activate purinergic P2 receptors and transient receptor potential (TRP) channels to effect changes in intracellular calcium. Despite the implied importance of ... More
Human methicillin-sensitive Staphylococcus aureus biofilms: potential associations with antibiotic resistance persistence and surface polysaccharide antigens.
AuthorsBabra C, Tiwari J, Costantino P, Sunagar R, Isloor S, Hegde N, Mukkur T
Journal
PubMed ID23686411
The development of persistent antibiotic resistance by human methicillin-sensitive Staphylococcus aureus (MSSA) strains and substantial association with poly-N-acetyl glucosamine (PNAG) in biofilms is reported in this investigation. Sixteen of 31 MSSA strains under study were found to have developed resistance to one or more antibiotics, with four strains, two of ... More