Identification and characterization of avian retroviruses in chicken embryo-derived yellow fever vaccines: investigation of transmission to vaccine recipients.
AuthorsHussain AI,Johnson JA,Da Silva Freire M,Heneine W
JournalJournal of virology
PubMed ID12502826
All currently licensed yellow fever (YF) vaccines are propagated in chicken embryos. Recent studies of chick cell-derived measles and mumps vaccines show evidence of two types of retrovirus particles, the endogenous avian retrovirus (EAV) and the endogenous avian leukosis virus (ALV-E), which originate from the chicken embryonic fibroblast substrates. In ... More
Genome-wide protective response used by group A Streptococcus to evade destruction by human polymorphonuclear leukocytes.
AuthorsVoyich JM, Sturdevant DE, Braughton KR, Kobayashi SD, Lei B, Virtaneva K, Dorward DW, Musser JM, DeLeo FR
JournalProc Natl Acad Sci U S A
PubMed ID12574517
Group A Streptococcus (GAS) evades polymorphonuclear leukocyte (PMN) phagocytosis and killing to cause human disease, including pharyngitis and necrotizing fasciitis (flesh-eating syndrome). We show that GAS genes differentially regulated during phagocytic interaction with human PMNs comprise a global pathogen-protective response to innate immunity. GAS prophage genes and genes involved in ... More
Group A Streptococcus gene expression in humans and cynomolgus macaques with acute pharyngitis.
AuthorsVirtaneva K, Graham MR, Porcella SF, Hoe NP, Su H, Graviss EA, Gardner TJ, Allison JE, Lemon WJ, Bailey JR, Parnell MJ, Musser JM
JournalInfect Immun
PubMed ID12654842
The molecular mechanisms used by group A Streptococcus (GAS) to survive on the host mucosal surface and cause acute pharyngitis are poorly understood. To provide new information about GAS host-pathogen interactions, we used real-time reverse transcription-PCR (RT-PCR) to analyze transcripts of 17 GAS genes in throat swab specimens taken from ... More
Rapid diagnosis of Ebola hemorrhagic fever by reverse transcription-PCR in an outbreak setting and assessment of patient viral load as a predictor of outcome.
AuthorsTowner JS, Rollin PE, Bausch DG, Sanchez A, Crary SM, Vincent M, Lee WF, Spiropoulou CF, Ksiazek TG, Lukwiya M, Kaducu F, Downing R, Nichol ST
JournalJ Virol
PubMed ID15047846
The largest outbreak on record of Ebola hemorrhagic fever (EHF) occurred in Uganda from August 2000 to January 2001. The outbreak was centered in the Gulu district of northern Uganda, with secondary transmission to other districts. After the initial diagnosis of Sudan ebolavirus by the National Institute for Virology in ... More
The HtrA protease from Streptococcus pneumoniae digests both denatured proteins and the competence-stimulating peptide.
AuthorsCassone M, Gagne AL, Spruce LA, Seeholzer SH, Sebert ME,
JournalJ Biol Chem
PubMed ID23012372
'The HtrA protease of Streptococcus pneumoniae functions both in a general stress response role and as an error sensor that specifically represses genetic competence when the overall level of biosynthetic errors in cellular proteins is low. However, the mechanism through which HtrA inhibits development of competence has been unknown. We ... More
Prevalence and quantitation of species C adenovirus DNA in human mucosal lymphocytes.
AuthorsGarnett CT, Erdman D, Xu W, Gooding LR
JournalJ Virol
PubMed ID12368303
'The common species C adenoviruses (serotypes Ad1, Ad2, Ad5, and Ad6) infect more than 80% of the human population early in life. Following primary infection, the virus can establish an asymptomatic persistent infection in which infectious virions are shed in feces for several years. The probable source of persistent virus ... More
Methylation-associated silencing of the thrombospondin-1 gene in human neuroblastoma.
AuthorsYang QW, Liu S, Tian Y, Salwen HR, Chlenski A, Weinstein J, Cohn SL
JournalCancer Res
PubMed ID14559817
'Tumor angiogenesis, a major requirement for tumor outgrowth and metastasis, is regulated by pro- and antiangiogenic factors. Methylation-associated inactivation of the angiogenesis inhibitor thrombospondin-1 (TSP-1) has been observed recently in some adult tumors. To investigate the role of TSP-1 in pediatric cancer, we examined its pattern of expression and mechanisms ... More
Quantitative detection of Moraxella catarrhalis in nasopharyngeal secretions by real-time PCR.
AuthorsGreiner O, Day PJ, Altwegg M, Nadal D
JournalJ Clin Microbiol
PubMed ID12682118
'The recognition of Moraxella catarrhalis as an important cause of respiratory tract infections has been protracted, mainly because it is a frequent commensal organism of the upper respiratory tract and the diagnostic sensitivity of blood or pleural fluid culture is low. Given that the amount of M. catarrhalis bacteria in ... More
Distinct molecular conformations of the estrogen receptor alpha complex exploited by environmental estrogens.
AuthorsBentrem D, Fox JE, Pearce ST, Liu H, Pappas S, Kupfer D, Zapf JW, Jordan VC
JournalCancer Res
PubMed ID14612550
'We have advanced the view that estrogens activate the estrogen receptor (ER) alpha complex differently. A group of planar (estradiol, genistein, and coumestrol) and nonplanar (methoxychlor and its mono- and didemethylated phenolic metabolites) environmental estrogens, which are all full estrogens in MCF-7 breast cancer cell proliferation assays, was shown to ... More
Synthesis and evaluation of phosphorescent oligonucleotide probes for hybridisation assays.
AuthorsO'Sullivan PJ, Burke M, Soini AE, Papkovsky DB
JournalNucleic Acids Res
PubMed ID12409473
'Monofunctional, p-isothiocyanatophenyl-derivatives of platinum (II)-coproporphyrin-I (PtCP-NCS) were evaluated as phosphorescent labelling reagents for synthetic oligonucleotides containing a 3''- or 5''-amino modification. Synthesis and purification conditions were optimised to generate high yields and purity of PtCP-labelled oligonucleotide probes. Phosphorescent properties of the PtCP label have been shown to be largely unaffected ... More
Short PNA molecular beacons for real-time PCR allelic discrimination of single nucleotide polymorphisms.
AuthorsPetersen K, Vogel U, Rockenbauer E, Nielsen KV, Kølvraa S, Bolund L, Nexø B
JournalMol Cell Probes
PubMed ID15051121
'The typing of a single nucleotide polymorphism with DNA probes is sometimes problematic because of the limited discriminating power of long DNA probes. As an alternative to existing assays, we have developed a real-time PCR assay for the genotyping of single nucleotide polymorphisms using short peptide nucleic acid (PNA) molecular ... More
Evaluation of the phosphorescent palladium(II)-coproporphyrin labels in separation-free hybridization assays.
AuthorsBurke M, O'Sullivan PJ, Soini AE, Berney H, Papkovsky DB
JournalAnal Biochem
PubMed ID12927834
'Palladium(II)-coproporphyrin label and a set of corresponding monofunctional labeling reagents with different linker arms were evaluated for labeling of oligonucleotides and subsequent use in hybridization assays. The properties of resulting oligonucleotide probes including phosphorescence spectra, quantum yields, lifetimes, and labeling yields were examined as functions of the label and oligonucleotide ... More
Development of an Alu-based, QSY 7-labeled primer PCR method for quantitation of human DNA in forensic samples.
AuthorsNicklas JA, Buel E
JournalJ Forensic Sci
PubMed ID12664984
'Determining the amount of human DNA extracted from a crime scene sample is an important step in DNA profiling. The forensic community relies almost entirely upon a technique (slot blot) to quantitate human DNA that is imprecise, time consuming, and labor intensive. This paper describes the development of a new ... More
The kinetics of aminosidine in renal patients with different degrees of renal failure.
AuthorsNovarini A, Montanari A, Bruschi G, Rossi E, Borghetti A, Migone L
JournalClin Nephrol
PubMed ID1157346
'The plasma concentration and urinary excretion after a single 500 mg dose of Aminosidine have been studied in 12 patients with different degrees of renal failure and 4 normal subjects. In normal subjects the plasma half-life is 2.47 hr; in patients with creatinine clearance (Ccr) of 30-60 ml/min, its 6.7 ... More
Rates of detection of Neisseria meningitidis in tonsils differ in relation to local incidence of invasive disease.
AuthorsGreiner O, Berger C, Day PJ, Meier G, Tang CM, Nadal D
JournalJ Clin Microbiol
PubMed ID12409352
'Nasopharyngeal swabbing substantially underestimates carriage of Neisseria meningitidis. Real-time PCR assays were employed to examine the presence of a broad range of bacteria and of N. meningitidis groups B and C, respectively, in tonsils from 26 individuals from Oxford, England, and 72 individuals from Zurich, Switzerland. The detection limit of ... More
Fluorescence excitation transfer immunoassay (FETI).
AuthorsUllman EF, Khanna PL
JournalMethods Enzymol
PubMed ID7033726
Quenching of biotinylated aequorin bioluminescence by dye-labeled avidin conjugates: application to homogeneous bioluminescence resonance energy transfer assays.
AuthorsAdamczyk M, Moore JA, Shreder K
JournalOrg Lett
PubMed ID11405714
[see reaction]. Avidin conjugates containing the covalently attached dyes QSY-7 and dabcyl were prepared and shown to quench the bioluminescence of biotinylated aequorin. Quenching efficiency was shown to be dependent on both the label-to-avidin ratio and the concentration of the avidin conjugate. These properties were exploited to develop a homogeneous ... More
Microarray analysis of the Mycobacterium tuberculosis transcriptional response to the acidic conditions found in phagosomes.
AuthorsFisher MA, Plikaytis BB, Shinnick TM
JournalJ Bacteriol
PubMed ID12081975
We used microarrays and real-time reverse transcription-PCR to analyze the global transcriptional response of Mycobacterium tuberculosis to low pH in vitro, which may mimic an environmental signal encountered by phagocytosed mycobacteria. Eighty-one genes were differentially expressed >1.5-fold, including many involved in fatty acid metabolism. The most highly induced genes showed ... More
Fluorescent-conjugated polymer superquenching facilitates highly sensitive detection of proteases.
AuthorsKumaraswamy S, Bergstedt T, Shi X, Rininsland F, Kushon S, Xia W, Ley K, Achyuthan K, McBranch D, Whitten D
JournalProc Natl Acad Sci U S A
PubMed ID15136731
Sensor formats have been developed for detecting the activity of proteolytic enzymes based on fluorescent conjugated polymer superquenching. These sensors employ a reactive peptide sequence within a tether linking a quencher to a biotin. The peptide binds to sensors containing colocated biotin-binding protein and fluorescent polymer by means of biotin-biotin ... More
Defective domain-domain interactions within the ryanodine receptor as a critical cause of diastolic Ca2+ leak in failing hearts.
AuthorsTateishi H, Yano M, Mochizuki M, Suetomi T, Ono M, Xu X, Uchinoumi H, Okuda S, Oda T, Kobayashi S, Yamamoto T, Ikeda Y, Ohkusa T, Ikemoto N, Matsuzaki M,
JournalCardiovasc Res
PubMed ID18996969
A domain peptide (DP) matching the Gly(2460)-Pro(2495) region of the cardiac type-2 ryanodine receptor (RyR2), DPc10, is known to mimic channel dysfunction associated with catecholaminergic polymorphic ventricular tachycardia (CPVT), owing to its interference in a normal interaction of the N-terminal (1-600) and central (2000-2500) domains (viz. domain unzipping). Using DPc10 ... More
Identification of target domains of the cardiac ryanodine receptor to correct channel disorder in failing hearts.
AuthorsYamamoto T, Yano M, Xu X, Uchinoumi H, Tateishi H, Mochizuki M, Oda T, Kobayashi S, Ikemoto N, Matsuzaki M,
JournalCirculation
PubMed ID18227387
We previously demonstrated that defective interdomain interaction between N-terminal (0 to 600) and central regions (2000 to 2500) of ryanodine receptor 2 (RyR2) induces Ca2+ leak in failing hearts and that K201 (JTV519) inhibits the Ca2+ leak by correcting the defective interdomain interaction. In the present report, we identified the ... More
Fluorophore-quencher based activatable targeted optical probes for detecting in vivo cancer metastases.
AuthorsOgawa M, Kosaka N, Longmire MR, Urano Y, Choyke PL, Kobayashi H,
JournalMol Pharm
PubMed ID19718793
In vivo molecularly targeted fluorescence imaging of tumors has been proposed as a strategy for improving cancer detection and management. Activatable fluorophores, which increased their fluorescence by 10-fold after binding tumor cells, result in much higher target to background ratios than conventional fluorophores. We developed an in vivo targeted activatable ... More
Visualizing the distribution and transport of mRNAs in living cells.
AuthorsBratu DP, Cha BJ, Mhlanga MM, Kramer FR, Tyagi S
JournalProc Natl Acad Sci U S A
PubMed ID14583593
We have visualized the movements of native mRNAs in living cells. Using nuclease-resistant molecular beacons, we imaged the transport and localization of oskar mRNA in Drosophila melanogaster oocytes. When the localization pattern was altered by genetic manipulation of the mRNA's 3' untranslated region, or by chemical perturbation of the intracellular ... More
Efficiencies of fluorescence resonance energy transfer and contact-mediated quenching in oligonucleotide probes.
AuthorsMarras SA, Kramer FR, Tyagi S
JournalNucleic Acids Res
PubMed ID12409481
An important consideration in the design of oligonucleotide probes for homogeneous hybridization assays is the efficiency of energy transfer between the fluorophore and quencher used to label the probes. We have determined the efficiency of energy transfer for a large number of combinations of commonly used fluorophores and quenchers. We ... More
Resonance energy transfer for assessing the molecular integrity of proteins for local delivery.
AuthorsWu D, Edelman ER
JournalBiotechnol Bioeng
PubMed ID14755558
It remains unclear whether the limitations to the therapeutic potential of angiogenic growth factors stem from pharmacokinetic concerns related to inadequate delivery or from a reduced sensitivity of target tissues. Here, we report a novel method using resonance energy transfer to assess the molecular integrity of proteins after local delivery. ... More
Novel immuno-FRET assay method for Bacillus spores and Escherichia coli O157:H7.
AuthorsBruno JG, Ulvick SJ, Uzzell GL, Tabb JS, Valdes ER, Batt CA
JournalBiochem Biophys Res Commun
PubMed ID11573946
Novel immunofluorescence resonance energy transfer (immuno-FRET) assays for both Bacillus cereus spores and Escherichia coli O157:H7 are reported. Both assays involve the use of dual (QSY-7 and Oregon Green 514-antibody)-labeled spores or vegetative bacteria, such that Oregon Green 514-labeled antibodies are quenched by proximal QSY-7 molecules that are covalently bound ... More
SPARC is a key Schwannian-derived inhibitor controlling neuroblastoma tumor angiogenesis.
AuthorsChlenski A, Liu S, Crawford SE, Volpert OV, DeVries GH, Evangelista A, Yang Q, Salwen HR, Farrer R, Bray J, Cohn SL
JournalCancer Res
PubMed ID12499280
Neuroblastoma (NB), a common pediatric neoplasm, consists of two main cell populations: neuroblastic/ganglionic cells and Schwann cells. NB tumors with abundant Schwannian stroma display a more benign clinical behavior than stroma-poor tumors. Recent studies suggest that Schwann cells influence NB tumor growth via secreted factors that induce differentiation, suppress proliferation, ... More
Actin filament uncapping localizes to ruffling lamellae and rocketing vesicles.
AuthorsAllen PG
JournalNat Cell Biol
PubMed ID14557819
Regulated actin filament assembly is critical for eukaryotic cell physiology. Actin filaments are polar structures, and those with free high affinity or barbed ends are crucial for actin dynamics and cell motility. Actin filament barbed-end-capping proteins inhibit filament elongation after binding, and their regulated disassociation is proposed to provide a ... More
A novel approach to the analysis of specificity, clonality, and frequency of HIV-specific T cell responses reveals a potential mechanism for control of viral escape.
AuthorsDouek DC, Betts MR, Brenchley JM, Hill BJ, Ambrozak DR, Ngai KL, Karandikar NJ, Casazza JP, Koup RA
JournalJ Immunol
PubMed ID11884484
Escape from the CD8(+) T cell response through epitope mutations can lead to loss of immune control of HIV replication. Theoretically, escape from CD8(+) T cell recognition is less likely when multiple TCRs target individual MHC/peptide complexes, thereby increasing the chance that amino acid changes in the epitope could be ... More
Fluorescence assay of SIRT protein deacetylases using an acetylated peptide substrate and a secondary trypsin reaction.
AuthorsMarcotte PA, Richardson PL, Richardson PR, Guo J, Barrett LW, Xu N, Gunasekera A, Glaser KB
JournalAnal Biochem
PubMed ID15301953
A novel fluorescent substrate was devised for the sirtuin (SIRT) class of human protein deacetylases comprised of a peptide sequence containing a single acetyl-lysine residue, with a fluorescent group (tetramethylrhodamine-6-carboxylic acid, 6-TAMRA) near the carboxyl terminus and a nonfluorescent quenching group (QSY-7) near the amino terminus. The peptide sequence is ... More
Human immunodeficiency virus type 1 neutralization measured by flow cytometric quantitation of single-round infection of primary human T cells.
AuthorsMascola JR, Louder MK, Winter C, Prabhakara R, De Rosa SC, Douek DC, Hill BJ, Gabuzda D, Roederer M
JournalJ Virol
PubMed ID11967298
There is currently intensive research on the design of novel human immunodeficiency virus type 1 (HIV-1) vaccine immunogens that can elicit potent neutralizing antibodies. A prerequisite for comparing and optimizing these strategies is the ability to precisely measure neutralizing antibody responses. To this end, we sought to develop an assay ... More
Beta-secretase cleavage at amino acid residue 34 in the amyloid beta peptide is dependent upon gamma-secretase activity.
AuthorsShi XP, Tugusheva K, Bruce JE, Lucka A, Wu GX, Chen-Dodson E, Price E, Li Y, Xu M, Huang Q, Sardana MK, Hazuda DJ
JournalJ Biol Chem
PubMed ID12665519
The amyloid beta peptides (Abeta) are the major components of the senile plaques characteristic of Alzheimer's disease. Abeta peptides are generated from the cleavage of amyloid precursor protein (APP) by beta- and gamma-secretases. Beta-secretase (BACE), a type-I transmembrane aspartyl protease, cleaves APP first to generate a 99-amino acid membrane-associated fragment ... More