JournalProceedings of the National Academy of Sciences of the United States of America
PubMed ID16754881
MicroRNAs (miRNAs) are endogenous noncoding RNAs, which negatively regulate gene expression. To determine genomewide miRNA DNA copy number abnormalities in cancer, 283 known human miRNA genes were analyzed by high-resolution array-based comparative genomic hybridization in 227 human ovarian cancer, breast cancer, and melanoma specimens. A high proportion of genomic loci ... More
A fluorescence-based assay for multisubunit DNA-dependent RNA polymerases.
AuthorsKuhlman P, Duff HL, Galant A
JournalAnal Biochem
PubMed ID14690681
The properties of DNA-dependent RNA polymerases have been studied since the 1960s, but considerable interest in probing RNA polymerase structure/function relationships, the roles of different classes of RNA polymerases in cellular processes, and the feasibility of using RNA polymerases as drug targets still exists. Historically, RNA polymerase activity has been ... More
Quantification of mRNA using real-time RT-PCR.
AuthorsNolan T, Hands RE, Bustin SA
JournalNat Protoc
PubMed ID17406449
'The real-time reverse transcription polymerase chain reaction (RT-qPCR) addresses the evident requirement for quantitative data analysis in molecular medicine, biotechnology, microbiology and diagnostics and has become the method of choice for the quantification of mRNA. Although it is often described as a "gold" standard, it is far from being a ... More
A systemic small RNA signaling system in plants.
AuthorsYoo BC, Kragler F, Varkonyi-Gasic E, Haywood V, Archer-Evans S, Lee YM, Lough TJ, Lucas WJ
JournalPlant Cell
PubMed ID15258266
'Systemic translocation of RNA exerts non-cell-autonomous control over plant development and defense. Long-distance delivery of mRNA has been proven, but transport of small interfering RNA and microRNA remains to be demonstrated. Analyses performed on phloem sap collected from a range of plants identified populations of small RNA species. The dynamic ... More
Fibrinogen induces IL-8 synthesis in human neutrophils stimulated with formyl-methionyl-leucyl-phenylalanine or leukotriene B(4).
AuthorsKuhns DB, Nelson EL, Alvord WG, Gallin JI
JournalJ Immunol
PubMed ID11509634
'Human exudative neutrophils have greatly increased stores of the neutrophil chemoattractant IL-8 compared with peripheral blood cells, but the mechanism for the increase is not defined. In this report, we show that treatment of peripheral blood neutrophils with the chemotactic peptide fMLP or with leukotriene B(4) or fibrinogen results in ... More
Gene expression changes in foam cells and the role of chemokine receptor CCR7 during atherosclerosis regression in ApoE-deficient mice.
AuthorsTrogan E, Feig JE, Dogan S, Rothblat GH, Angeli V, Tacke F, Randolph GJ, Fisher EA
JournalProc Natl Acad Sci U S A
PubMed ID16537455
'Atherosclerosis regression is an important clinical goal. In previous studies of regression in mice, the rapid loss of plaque foam cells was explained by emigration to lymph nodes, a process reminiscent of dendritic cells. In the present study, plaque-containing arterial segments from apoE-/- mice were transplanted into WT recipient normolipidemic ... More
Accurately quantifying low-abundant targets amid similar sequences by revealing hidden correlations in oligonucleotide microarray data.
AuthorsMarcelino LA, Backman V, Donaldson A, Steadman C, Thompson JR, Preheim SP, Lien C, Lim E, Veneziano D, Polz MF
JournalProc Natl Acad Sci U S A
PubMed ID16950880
'Microarrays have enabled the determination of how thousands of genes are expressed to coordinate function within single organisms. Yet applications to natural or engineered communities where different organisms interact to produce complex properties are hampered by theoretical and technological limitations. Here we describe a general method to accurately identify low-abundant ... More
Quantification of enterovirus RNA in sludge samples using single tube real-time RT-PCR.
AuthorsMonpoeho S, Dehée A, Mignotte B, Schwartzbrod L, Marechal V, Nicolas JC, Billaudel S, Férré V
JournalBiotechniques
PubMed ID10907082
'We have developed a quantitative RT-PCR method that can be used to determine the amount of enterovirus RNA in urban sludge samples. This method combines Taq-Man technology with the ABI Prism 7700 real-time sequence detection system. We optimized a one-step RT-PCR that uses a dual-labeled fluorogenic probe to quantify the ... More
Ca2+-dependent production and release of IL-8 in human neutrophils.
AuthorsKuhns DB, Young HA, Gallin EK, Gallin JI
JournalJ Immunol
PubMed ID9780210
'IL-8, a potent neutrophil chemoattractant that is elevated about 200-fold in exudative neutrophils isolated from localized inflammatory sites in vivo, is thought to play a major role in recruitment of neutrophils to inflammatory sites. Incubation of peripheral blood neutrophils with thapsigargin, an inhibitor of the endoplasmic reticulum Ca2+-sequestering-ATPase, causes a ... More
mRNA expression profiling of laser microbeam microdissected cells from slender embryonic structures.
AuthorsScheidl SJ, Nilsson S, Kalén M, Hellström M, Takemoto M, Håkansson J, Lindahl P
JournalAm J Pathol
PubMed ID11891179
'Microarray hybridization has rapidly evolved as an important tool for genomic studies and studies of gene regulation at the transcriptome level. Expression profiles from homogenous samples such as yeast and mammalian cell cultures are currently extending our understanding of biology, whereas analyses of multicellular organisms are more difficult because of ... More
Stimulation of HIV-1 minus strand strong stop DNA transfer by genomic sequences 3' of the primer binding site.
AuthorsSong M, Balakrishnan M, Chen Y, Roques BP, Bambara RA
JournalJ Biol Chem
PubMed ID16782713
'The mechanism of human immunodeficiency virus 1 (HIV-1) minus strand transfer was examined using a genomic RNA sequence-based donor-acceptor template system. The donor RNA, D199, was a 199-nucleotide sequence from the 5''-end of the genome to the primer binding site (PBS) and shared 97 nucleotides of homology with the acceptor ... More
alpha3alpha5beta2-Nicotinic acetylcholine receptor contributes to the wound repair of the respiratory epithelium by modulating intracellular calcium in migrating cells.
AuthorsTournier JM, Maouche K, Coraux C, Zahm JM, Cloëz-Tayarani I, Nawrocki-Raby B, Bonnomet A, Burlet H, Lebargy F, Polette M, Birembaut P
JournalAm J Pathol
PubMed ID16400009
'Nicotinic acetylcholine receptors (nAChRs), present in human bronchial epithelial cells (HBECs), have been shown in vitro to modulate cell shape. Because cell spreading and migration are important mechanisms involved in the repair of the bronchial epithelium, we investigated the potential role of nAChRs in the wound repair of the bronchial ... More
An evaluation of the performance of cDNA microarrays for detecting changes in global mRNA expression.
AuthorsYue H, Eastman PS, Wang BB, Minor J, Doctolero MH, Nuttall RL, Stack R, Becker JW, Montgomery JR, Vainer M, Johnston R
JournalNucleic Acids Res
PubMed ID11292855
'The cDNA microarray is one technological approach that has the potential to accurately measure changes in global mRNA expression levels. We report an assessment of an optimized cDNA microarray platform to generate accurate, precise and reliable data consistent with the objective of using microarrays as an acquisition platform to populate ... More
A mitotic cascade of NIMA family kinases. Nercc1/Nek9 activates the Nek6 and Nek7 kinases.
AuthorsBelham C, Roig J, Caldwell JA, Aoyama Y, Kemp BE, Comb M, Avruch J
JournalJ Biol Chem
PubMed ID12840024
'The Nek family of protein kinases in humans is composed of 11 members that share an amino-terminal catalytic domain related to NIMA, an Aspergillus kinase involved in the control of several aspects of mitosis, and divergent carboxyl-terminal tails of varying length. Nek6 (314AA) and Nek7 (303AA), 76% identical, have little ... More
Interleukin-15 rescues tolerant CD8+ T cells for use in adoptive immunotherapy of established tumors.
'CD8+ T cells can mediate eradication of established tumors, and strategies to amplify tumor-reactive T-cell numbers by immunization or ex vivo expansion followed by adoptive transfer are currently being explored in individuals with cancer. Generating effective CD8+ T cell-mediated responses to tumors is often impeded by T-cell tolerance to relevant ... More
Anisotropic mechanosensing by mesenchymal stem cells.
AuthorsKurpinski K, Chu J, Hashi C, Li S
JournalProc Natl Acad Sci U S A
PubMed ID17060641
'Mesenchymal stem cells (MSCs) are a potential source for the construction of tissue-engineered vascular grafts. However, how vascular mechanical forces regulate the genetic reprogramming in MSCs is not well understood. Mechanical strain in the vascular wall is anisotropic and mainly in the circumferential direction. We have shown that cyclic uniaxial ... More
A preservation method that allows recovery of intact RNA from tissues dissected by laser capture microdissection.
AuthorsParlato R, Rosica A, Cuccurullo V, Mansi L, Macchia P, Owens JD, Mushinski JF, De Felice M, Bonner RF, Di Lauro R
JournalAnal Biochem
PubMed ID11779104
'We report a novel method for preparing samples for laser capture microdissection. The procedure described here permits extraction of intact RNA while preserving morphology, thus being suitable both for identification of specific cells and for analysis of their gene expression. The method is applicable to both mouse embryos and human ... More
Comparison of RNA assay methods used to normalize cDNA for quantitative real-time PCR.
AuthorsLee EJ, Schmittgen TD
JournalAnal Biochem
PubMed ID16930526
Quantification of cDNA generated by reverse transcription of total RNA provides a simple alternative tool for quantitative RT-PCR normalization.
AuthorsLibus J, Storchová H
JournalBiotechniques
PubMed ID16925017
Comparison of RiboGreen and 18S rRNA quantitation for normalizing real-time RT-PCR expression analysis.
AuthorsHashimoto JG, Beadles-Bohling AS, Wiren KM,
JournalBiotechniques
PubMed ID14740484
None
Multitasking with molecular dynamics Typhoon: quantifying nucleic acids and autoradiographs.
AuthorsGoodrich RJ, Ostermeier GC, Krawetz SA
JournalBiotechnol Lett
PubMed ID12889815
With increased sensitivity and specificity, fluorescent assays are rapidly becoming the method of choice for nucleic acid quantification. The utility of the Typhoon scanner has now been extended to accurately measure low levels of DNA and RNA (5 ng ml(-1)) with PicoGreen and RiboGreen dyes. In addition, with a few ... More
A sensitive and high-throughput assay to detect low-abundance proteins in serum.
AuthorsZhang H, Cheng X, Richter M, Greene MI
JournalNat Med
PubMed ID16532003
The ability to detect antigens immunologically is limited by the affinity of the antibodies and the amount of antigens. We have now succeeded in creating a modular, facile amplification system, termed fluorescent amplification catalyzed by T7 polymerase technique (FACTT). Such a system can detect protein targets specifically at subfemtomolar levels ... More
Gene expression analysis by transcript profiling coupled to a gene database query.
AuthorsShimkets RA, Lowe DG, Tai JT, Sehl P, Jin H, Yang R, Predki PF, Rothberg BE, Murtha MT, Roth ME, Shenoy SG, Windemuth A, Simpson JW, Simons JF, Daley MP, Gold SA, McKenna MP, Hillan K, Went GT, Rothberg JM
JournalNat Biotechnol
PubMed ID10429247
We describe an mRNA profiling technique for determining differential gene expression that utilizes, but does not require, prior knowledge of gene sequences. This method permits high-throughput reproducible detection of most expressed sequences with a sensitivity of greater than 1 part in 100,000. Gene identification by database query of a restriction ... More
Oral cancer in vivo gene expression profiling assisted by laser capture microdissection and microarray analysis.
AuthorsAlevizos I, Mahadevappa M, Zhang X, Ohyama H, Kohno Y, Posner M, Gallagher GT, Varvares M, Cohen D, Kim D, Kent R, Donoff RB, Todd R, Yung CM, Warrington JA, Wong DT
JournalOncogene
PubMed ID11593428
Large scale gene expression profiling was carried out on laser capture microdissected (LCM) tumor and normal oral epithelial cells and analysed on high-density oligonucleotide microarrays. About 600 genes were found to be oral cancer associated. These oral cancer associated genes include oncogenes, tumor suppressors, transcription factors, xenobiotic enzymes, metastatic proteins, ... More
Mesenchymal cells stimulate capillary morphogenesis via distinct proteolytic mechanisms.
AuthorsGhajar CM, Kachgal S, Kniazeva E, Mori H, Costes SV, George SC, Putnam AJ,
JournalExp Cell Res
PubMed ID20067788
During angiogenesis, endothelial cells (ECs) degrade their surrounding extracellular matrix (ECM) to facilitate invasion. How interactions between ECs and other cells within their microenvironment facilitate this process is only partially understood. We have utilized a tractable 3D co-culture model to investigate the proteolytic mechanisms by which pre-committed or more highly ... More
Sequence-independent and reversible photocontrol of transcription/expression systems using a photosensitive nucleic acid binder.
AuthorsEstévez-Torres A, Crozatier C, Diguet A, Hara T, Saito H, Yoshikawa K, Baigl D,
JournalProc Natl Acad Sci U S A
PubMed ID19617550
To understand non-trivial biological functions, it is crucial to develop minimal synthetic models that capture their basic features. Here, we demonstrate a sequence-independent, reversible control of transcription and gene expression using a photosensitive nucleic acid binder (pNAB). By introducing a pNAB whose affinity for nucleic acids is tuned by light, ... More
Transcriptome analysis of proliferating Arabidopsis endosperm reveals biological implications for the control of syncytial division, cytokinin signaling, and gene expression regulation.
During the early stages of seed development, Arabidopsis (Arabidopsis thaliana) endosperm is syncytial and proliferates rapidly through repeated rounds of mitosis without cytokinesis. This stage of endosperm development is important in determining final seed size and is a model for studying aspects of cellular and molecular biology, such as the ... More
A gnotobiotic transgenic mouse model for studying interactions between small intestinal enterocytes and intraepithelial lymphocytes.
AuthorsMysorekar IU, Lorenz RG, Gordon JI,
JournalJ Biol Chem
PubMed ID12138109
The mouse intestinal epithelium undergoes continuous renewal throughout life. Intraepithelial lymphocytes (IELs) represent a significant fraction of this epithelium and play an important role in intestinal mucosal barrier function. We have generated a germ-free transgenic mouse model to examine the effects of a genetically engineered proliferative abnormality in the principal ... More
Hedgehog signalling is essential for maintenance of cancer stem cells in myeloid leukaemia.
AuthorsZhao C, Chen A, Jamieson CH, Fereshteh M, Abrahamsson A, Blum J, Kwon HY, Kim J, Chute JP, Rizzieri D, Munchhof M, VanArsdale T, Beachy PA, Reya T,
JournalNature
PubMed ID19169242
Although the role of Hedgehog (Hh) signalling in embryonic pattern formation is well established, its functions in adult tissue renewal and maintenance remain unclear, and the relationship of these functions to cancer development has not been determined. Here we show that the loss of Smoothened (Smo), an essential component of ... More
Therapeutic suppression of translation initiation factor eIF4E expression reduces tumor growth without toxicity.
AuthorsGraff JR, Konicek BW, Vincent TM, Lynch RL, Monteith D, Weir SN, Schwier P, Capen A, Goode RL, Dowless MS, Chen Y, Zhang H, Sissons S, Cox K, McNulty AM, Parsons SH, Wang T, Sams L, Geeganage S, Douglass LE, Neubauer BL, Dean NM, Blanchard K, Shou J, Stancato LF, Carter JH, Marcusson EG,
JournalJ Clin Invest
PubMed ID17786246
Expression of eukaryotic translation initiation factor 4E (eIF4E) is commonly elevated in human and experimental cancers, promoting angiogenesis and tumor growth. Elevated eIF4E levels selectively increase translation of growth factors important in malignancy (e.g., VEGF, cyclin D1) and is thereby an attractive anticancer therapeutic target. Yet to date, no eIF4E-specific ... More
MicroRNA-143 regulates adipocyte differentiation.
AuthorsEsau C, Kang X, Peralta E, Hanson E, Marcusson EG, Ravichandran LV, Sun Y, Koo S, Perera RJ, Jain R, Dean NM, Freier SM, Bennett CF, Lollo B, Griffey R
JournalJ Biol Chem
PubMed ID15504739
MicroRNAs (miRNAs) are endogenously expressed 20-24 nucleotide RNAs thought to repress protein translation through binding to a target mRNA (1-3). Only a few of the more than 250 predicted human miRNAs have been assigned any biological function. In an effort to uncover miRNAs important during adipocyte differentiation, antisense oligonucleotides (ASOs) ... More
Positional effect of chemical modifications on short interference RNA activity in mammalian cells.
AuthorsPrakash TP, Allerson CR, Dande P, Vickers TA, Sioufi N, Jarres R, Baker BF, Swayze EE, Griffey RH, Bhat B
JournalJ Med Chem
PubMed ID15974578
A systematic study on the effect of 2'-sugar modifications (2'-F (2'-F-2'-deoxy-nucleoside residues), 2'-O-Me (2'-O-methyl-nucleoside residues), and 2'-O-MOE [2'-O-(2-methoxyethyl)]-nucleoside residues) in the antisense and sense strands of short interference RNA (siRNA) was performed in HeLa cells. The study of the antisense strand of siRNAs demonstrated that activity depends on the position ... More
Human alveolar macrophages are deficient in PTEN. The role of endogenous oxidants.
AuthorsFlaherty DM, Monick MM, Hinde SL
JournalJ Biol Chem
PubMed ID16371363
Human alveolar macrophages play a critical role in host defense and in the development of inflammation and fibrosis in the lung. Unlike their precursor cells, blood monocytes, alveolar macrophages are long-lived and tend to be resistant to apoptotic stimuli. In this study, we examined the role of differentiation in altering ... More
Use of biotin-labeled nucleic acids for protein purification and agarose-based chemiluminescent electromobility shift assays.
AuthorsRodgers JT, Patel P, Hennes JL, Bolognia SL, Mascotti DP
JournalAnal Biochem
PubMed ID10625515
We have employed biotin-labeled RNA to serve two functions. In one, the biotin tethers the RNA to streptavidin-agarose beads, creating an affinity resin for protein purification. In the other, the biotin functions as a label for use in a modified chemiluminescent electromobility shift assay (EMSA), a technique used to detect ... More
Same-sample analysis of ethoxyresorufin-O-deethylase activity and cytochrome P4501A mRNA abundance in chicken embryo hepatocytes.
AuthorsHead JA, Kennedy SW
JournalAnal Biochem
PubMed ID17137551
Inducibility of the cytochrome P4501A4 (CYP1A4) enzyme, measured as ethoxyresorufin-O-deethylase (EROD) activity, has been used as a biomarker for sensitivity to the effects of dioxin-like compounds in avian species. Here, we present a quantitative reverse transcriptase-PCR (Q-PCR) method for assessing this biomarker response at the level of messenger RNA (mRNA) ... More
The Orphan G protein-coupled receptors GPR41 and GPR43 are activated by propionate and other short chain carboxylic acids.
AuthorsBrown AJ, Goldsworthy SM, Barnes AA, Eilert MM, Tcheang L, Daniels D, Muir AI, Wigglesworth MJ, Kinghorn I, Fraser NJ, Pike NB, Strum JC, Steplewski KM, Murdock PR, Holder JC, Marshall FH, Szekeres PG, Wilson S, Ignar DM, Foord SM, Wise A, Dowell SJ
JournalJ Biol Chem
PubMed ID12496283
GPR41 and GPR43 are related members of a homologous family of orphan G protein-coupled receptors that are tandemly encoded at a single chromosomal locus in both humans and mice. We identified the acetate anion as an agonist of human GPR43 during routine ligand bank screening in yeast. This activity was ... More
RNA quantitation by fluorescence-based solution assay: RiboGreen reagent characterization.
AuthorsJones LJ, Yue ST, Cheung CY, Singer VL
JournalAnal Biochem
PubMed ID9882416
We describe the development of a sensitive fluorescence-based solution assay for RNA using a new dye, RiboGreen RNA quantitation reagent. RiboGreen reagent exhibits >1000-fold fluorescence enhancement and high quantum yield (0.65) upon binding nucleic acids, with excitation and emission maxima near those of fluorescein. Unbound dye is essentially nonfluorescent and ... More
Performance of the translational apparatus varies with the ecological strategies of bacteria.
AuthorsDethlefsen L, Schmidt TM
JournalJ Bacteriol
PubMed ID17277058
Protein synthesis is the predominant activity of growing bacteria; the protein synthesis system accounts for more than one-half the cell's dry mass and consumes most of the cell's energy during rapid growth. Translation has been studied extensively using model organisms, and the translational apparatus is qualitatively similar in terms of ... More
Binding of fluorescent dye to genomic RNA inside intact human rhinovirus after viral capsid penetration investigated by capillary electrophoresis.
AuthorsKremser L, Okun VM, Nicodemou A, Blaas D, Kenndler E
JournalAnal Chem
PubMed ID14961716
RiboGreen is used for concentration measurements of RNA. Upon binding to the RNA, an approximately 1000-fold increase in sensitivity in comparison with the UV absorbance of the free polynucleotide is observed. In the present work, we demonstrate that this dye can penetrate in a time- and temperature-dependent manner the intact ... More
Phosphophoryn regulates the gene expression and differentiation of NIH3T3, MC3T3-E1, and human mesenchymal stem cells via the integrin/MAPK signaling pathway.
AuthorsJadlowiec J, Koch H, Zhang X, Campbell PG, Seyedain M, Sfeir C
JournalJ Biol Chem
PubMed ID15371433
Extracellular matrix proteins (ECMs) serve as both a structural support for cells and a dynamic biochemical network that directs cellular activities. ECM proteins such as those of the SIBLING family (small integrin-binding ligand glycoprotein) could possess inherent growth factor activity. In this study, we demonstrate that exon 5 of dentin ... More
Molecular mechanism of glutathione-mediated protection from oxidized low-density lipoprotein-induced cell injury in human macrophages: role of glutathione reductase and glutaredoxin.
AuthorsWang Y, Qiao M, Mieyal JJ, Asmis LM, Asmis R
JournalFree Radic Biol Med
PubMed ID16895798
Macrophage death is a hallmark of advanced atherosclerotic plaque, and oxidized low-density lipoprotein (OxLDL) found in these lesions is believed to contribute to macrophage injury. However, the underlying mechanisms of this phenomenon are only poorly understood. Here we show that in human monocyte-derived macrophages, OxLDL depleted intracellular glutathione (GSH) and ... More
Distinct roles for nucleic acid in in vitro assembly of purified Mason-Pfizer monkey virus CANC proteins.
AuthorsUlbrich P, Haubova S, Nermut MV, Hunter E, Rumlova M, Ruml T
JournalJ Virol
PubMed ID16809314
In contrast to other retroviruses, Mason-Pfizer monkey virus (M-PMV) assembles immature capsids in the cytoplasm. We have compared the ability of minimal assembly-competent domains from M-PMV and human immunodeficiency virus type 1 (HIV-1) to assemble in vitro into virus-like particles in the presence and absence of nucleic acids. A fusion ... More
Low abundance of telomerase in yeast: implications for telomerase haploinsufficiency.
AuthorsMozdy AD, Cech TR
JournalRNA
PubMed ID16894218
Telomerase is an RNA-dependent reverse transcriptase that maintains telomeric DNA at a species-specific equilibrium length. To determine an upper limit for the number of telomerase molecules in a Saccharomyces cerevisiae cell, we have established real-time RT-PCR assays to quantify the noncoding telomerase RNA, TLC1. We find that the number of ... More
Control of adipose triglyceride lipase action by serine 517 of perilipin A globally regulates protein kinase A-stimulated lipolysis in adipocytes.
Phosphorylation of the lipid droplet-associated protein perilipin A (Peri A) mediates the actions of cyclic AMP-dependent protein kinase A (PKA) to stimulate triglyceride hydrolysis (lipolysis) in adipocytes. Studies addressing how Peri A PKA sites regulate adipocyte lipolysis have relied on non-adipocyte cell models, which express neither adipose triglyceride lipase (ATGL), ... More
Mammalian class Sigma glutathione S-transferases: catalytic properties and tissue-specific expression of human and rat GSH-dependent prostaglandin D2 synthases.
AuthorsJowsey IR, Thomson AM, Flanagan JU, Murdock PR, Moore GB, Meyer DJ, Murphy GJ, Smith SA, Hayes JD
JournalBiochem J
PubMed ID11672424
GSH-dependent prostaglandin D(2) synthase (PGDS) enzymes represent the only vertebrate members of class Sigma glutathione S-transferases (GSTs) identified to date. Complementary DNA clones encoding the orthologous human and rat GSH-dependent PGDS (hPGDS and rPGDS, respectively) have been expressed in Escherichia coli, and the recombinant proteins isolated by affinity chromatography. The ... More
Cytochrome P450 gene induction in rats ex vivo assessed by quantitative real-time reverse transcriptase-polymerase chain reaction (TaqMan).
AuthorsBaldwin SJ, Bramhall JL, Ashby CA, Yue L, Murdock PR, Hood SR, Ayrton AD, Clarke SE
JournalDrug Metab Dispos
PubMed ID16531474
Drug-induced changes in expression of cytochrome P450 (P450) genes are a significant issue in the preclinical development of pharmaceuticals. For example, preclinically, P450 induction can affect safety studies by reducing the systemic exposure of a compound undergoing toxicological evaluation, thus limiting the exposure that can be safely investigated in patients. ... More
Pdx-1 links histone H3-Lys-4 methylation to RNA polymerase II elongation during activation of insulin transcription.
Expression of the insulin gene is nearly exclusive to the beta cells of the pancreatic islets. Although the sequence-specific transcription factors that regulate insulin expression have been well studied, the interrelationship between these factors, chromatin structure, and transcriptional elongation by RNA polymerase II (pol II) has remained undefined. In this ... More
The hyperglycemia-induced inflammatory response in adipocytes: the role of reactive oxygen species.
AuthorsLin Y, Berg AH, Iyengar P, Lam TK, Giacca A, Combs TP, Rajala MW, Du X, Rollman B, Li W, Hawkins M, Barzilai N, Rhodes CJ, Fantus IG, Brownlee M, Scherer PE
JournalJ Biol Chem
PubMed ID15536073
Hyperglycemia is a major independent risk factor for diabetic macrovascular disease. The consequences of exposure of endothelial cells to hyperglycemia are well established. However, little is known about how adipocytes respond to both acute as well as chronic exposure to physiological levels of hyperglycemia. Here, we analyze adipocytes exposed to ... More
Transcriptome reprogramming by cancer exosomes: identification of novel molecular targets in matrix and immune modulation.
AuthorsQadir F, Aziz MA, Sari CP, Ma H, Dai H, Wang X, Raithatha D, Da Silva LGL, Hussain M, Poorkasreiy SP, Hutchison IL, Waseem A, Teh MT
JournalMol Cancer
PubMed ID30008265
'Exosomes are extracellular vesicles released by almost all cell types, including cancer cells, into bodily fluids such as saliva, plasma, breast milk, semen, urine, cerebrospinal fluid, amniotic fluid, synovial fluid and sputum. Their key function being intercellular communication with both neighbouring as well as distant cells. Cancer exosomes have been ... More
Customizable Lipid Nanoparticle Materials for the Delivery of siRNAs and mRNAs.
'RNAs are a promising class of therapeutics given their ability to regulate protein concentrations at the cellular level. Developing safe and effective strategies to deliver RNAs remains important for realizing their full clinical potential. Here, we develop lipid nanoparticle formulations that can deliver short interfering RNAs (for gene silencing) or ... More
Early development of tendinopathy in humans: Sequence of pathological changes in structure and tissue turnover signaling.
'Overloading of tendon tissue with resulting chronic pain (tendinopathy) is a common disorder in occupational-, leisure- and sports-activity, but its pathogenesis remains poorly understood. To investigate the very early phase of tendinopathy, Achilles and patellar tendons were investigated in 200 physically active patients and 50 healthy control persons. Patients were ... More
3D microfluidic ex vivo culture of organotypic tumor spheroids to model immune checkpoint blockade.
AuthorsAref AR, Campisi M, Ivanova E, Portell A, Larios D, Piel BP, Mathur N, Zhou C, Coakley RV, Bartels A, Bowden M, Herbert Z, Hill S, Gilhooley S, Carter J, Cañadas I, Thai TC, Kitajima S, Chiono V, Paweletz CP, Barbie DA, Kamm RD, Jenkins RW
JournalLab Chip
PubMed ID30183789
Microfluidic culture has the potential to revolutionize cancer diagnosis and therapy. Indeed, several microdevices are being developed specifically for clinical use to test novel cancer therapeutics. To be effective, these platforms need to replicate the continuous interactions that exist between tumor cells and non-tumor cell elements of the tumor microenvironment ... More
Exosome-delivered microRNAs promote IFN-a secretion by human plasmacytoid DCs via TLR7.
AuthorsSalvi V, Gianello V, Busatto S, Bergese P, Andreoli L, D'Oro U, Zingoni A, Tincani A, Sozzani S, Bosisio D
JournalJCI Insight
PubMed ID29769437
The excessive production of type I IFNs is a hallmark and a main pathogenic mechanism of many autoimmune diseases, including systemic lupus erythematosus (SLE). In these pathologies, the sustained secretion of type I IFNs is dependent on the improper activation of plasmacytoid DCs (pDCs) by self-nucleic acids. However, the nature ... More
Extracellular microRNAs in blood differentiate between ischaemic and haemorrhagic stroke subtypes.
AuthorsKalani MYS, Alsop E, Meechoovet B, Beecroft T, Agrawal K, Whitsett TG, Huentelman MJ, Spetzler RF, Nakaji P, Kim S, Van Keuren-Jensen K
JournalJ Extracell Vesicles
PubMed ID32128071
Rapid identification of patients suffering from cerebral ischaemia, while excluding intracerebral haemorrhage, can assist with patient triage and expand patient access to chemical and mechanical revascularization. We sought to identify blood-based, extracellular microRNAs 15 (ex-miRNAs) derived from extracellular vesicles associated with major stroke subtypes using clinical samples from subjects with ... More