Publications

TitleAbstractYear(sorted descending)
Filter
PMID
Filter
genotoxicity of sediment extracts of the berre lagoon (france).to evaluate the genotoxic risk that contaminated sediment could constitute for benthic organisms, three contaminated (va, vc and vn) and one uncontaminated (rn) sediment samples were collected in the berre lagoon (france). potentially bioavailable contaminants in sediments were obtained using sediment extraction with synthetic seawater adjusted to ph 4 or ph 6, simulating the range of ph prevailing in the digestive tract of benthic organisms. the genotoxic activities of these extracts were evalu ...201222487561
micropatterning of mammalian cells on inorganic-based nanosponges.developing artificial scaffolding structures in vitro in order to mimic physiological-relevant situations in vivo is critical in many biological and medical arenas including bone and cartilage generation, biomaterials, small-scale biomedical devices, tissue engineering, as well as the development of nanofabrication methods. we focus on using simple physical principles (photolithography) and chemical techniques (liquid vapor deposition) to build non-cytotoxic scaffolds with a nanometer resolution ...201222483244
the bisecting glcnac in cell growth control and tumor progression.the bisecting glcnac is transferred to the core mannose residue of complex or hybrid n-glycans on glycoproteins by the β1,4-n-acetylglucosaminyltransferase iii (glcnact-iii) or mgat3. the addition of the bisecting glcnac confers unique lectin recognition properties to n-glycans. thus, lec10 gain-of-function chinese hamster ovary (cho) cells selected for the acquisition of ricin resistance, carry n-glycans with a bisecting glcnac, which enhances the binding of the erythroagglutinin e-pha, but red ...201222476631
identification of selective inhibitors of the potassium channel kv1.1-1.2((3)) by high-throughput virtual screening and automated patch clamp.two voltage-dependent potassium channels, kv1.1 (kcna1) and kv1.2 (kcna2), are found to co-localize at the juxtaparanodal region of axons throughout the nervous system and are known to co-assemble in heteromultimeric channels, most likely in the form of the concatemer kv1.1-1.2((3)) . loss of the myelin sheath, as is observed in multiple sclerosis, uncovers the juxtaparanodal region of nodes of ranvier in myelinated axons leading to potassium conductance, resulting in loss of nerve conduction. t ...201222473914
probing of c-terminal lysine variation in a recombinant monoclonal antibody production using chinese hamster ovary cells with chemically defined media.c-terminal lysine (c-k) variants are commonly observed in therapeutic monoclonal antibodies and recombinant proteins. heterogeneity of c-k residues is believed to result from varying degree of proteolysis by endogenous carboxypeptidase(s) during cell culture production. the achievement of batch-to-batch culture performance and product quality reproducibility is a key cell culture development criterion. understanding the operational parameters affecting c-k levels provides valuable insight into t ...201222473810
fenofibrate down-regulates renal oct2-mediated organic cation transport via pparα-independent pathways.fibrate drugs, the peroxisome proliferator-activated receptor alpha (pparα) agonists, are widely prescribed for the treatment of hyperlipidemia. the present study examined the effect of fibrate drugs on renal oct2 activity in a heterologous cell system [chinese hamster ovary (cho-k1) cells stably transfected with rabbit (rb) oct2], llc-pk1, and intact mouse renal cortical slices. we found that both in the cho-k1 cells expressing rboct2 and in llc-pk1 cells, fenofibrate significantly inhibited [³ ...201222473497
effects of decabrominated diphenyl ether (pbde-209) in regulation of growth and apoptosis of breast, ovarian, and cervical cancer cells.polybrominated diphenyl ethers (pbdes), commonly used in building materials, electronics, plastics, polyurethane foams, and textiles, are health hazards found in the environment.201222472210
different matrix attachment regions flanking a transgene effectively enhance gene expression in stably transfected chinese hamster ovary cells.numerous matrix attachment regions (mars) have been used to improve transgene expression in genetic engineering, but an efficient and stable expression vector is lacking. in the present study, a vector named pccf containing chloramphenicol acetyltransferase (cat) reporter gene cassettes was constructed. the cassettes were flanked by a β-interferon mar at the 5' upstream of the reporter gene cassettes, and a β-globin mar at the 3' site. after transfecting pccf into chinese hamster ovary cells, th ...201222472079
in silico identification of potential cholestasis-inducing agents via modeling of na(+)-dependent taurocholate cotransporting polypeptide substrate specificity.na(+)-dependent taurocholate cotransporting polypeptide (ntcp, slc10a1) is the main transporter facilitating the hepatic uptake of bile acids from the circulation. consequently, the interaction of xenobiotics, including therapeutic drugs, with the bile acid binding pocket of ntcp could lead to impairment of hepatic bile acid uptake. we pursued a 3d-pharmacophore approach to model the ntcp substrate and inhibitor specificity and investigated whether it is possible to identify compounds with intri ...201222641618
farnesyl pyrophosphate is an endogenous antagonist to adp-stimulated p2y₁₂ receptor-mediated platelet aggregation.farnesyl pyrophosphate (fpp) is an intermediate in cholesterol biosynthesis, and it has also been reported to activate platelet lpa (lysophosphatidic acid) receptors. the aim of this study was to investigate the role of extracellular fpp in platelet aggregation. human platelets were studied with light transmission aggregometry, flow cytometry and [³⁵s]gtpγs binding assays. as shown previously, fpp could potentiate lpa-stimulated shape change. surprisingly, fpp also acted as a selective insurmoun ...201222628078
rapamycin treatment inhibits cho cell death in a serum-free suspension culture by autophagy induction.rapamycin, a specific mtor inhibitor, has been used as a chemical activator in autophagy research both in vitro and in vivo. recently, autophagy has received attention as an anti-cell death engineering target in addition to apoptosis in the chinese hamster ovary (cho) cell engineering field. here, the effect of rapamycin and the subsequent autophagy induction is investigated on two cho cell lines, dg44 host and an antibody-producing recombinant cho (rcho), in a serum-free suspension culture. in ...201222627923
albumin 3'untranslated region facilitates increased recombinant protein production from chinese hamster ovary cells.chinese hamster ovary (cho) cells are used for recombinant protein production in the pharmaceutical industry but there is a need to improve expression levels. in the present work experiments were carried out to test the effectiveness of different 3'untranslated regions (3'utrs) in promoting production of a naturally secreted luciferase. seamless cloning was used to produce expression vectors in which gaussia princeps luciferase coding sequences were linked to the human albumin, immunoglobulin or ...201222623341
studying host cell protein interactions with monoclonal antibodies using high throughput protein a chromatography.protein a chromatography is typically used as the initial capture step in the purification of monoclonal antibodies produced in chinese hamster ovary (cho) cells. although exploiting an affinity interaction for purification, the level of host cell proteins in the protein a eluent varies significantly with different feedstocks. using a batch binding chromatography method, we performed a controlled study to assess host cell protein clearance across both mabselect sure and prosep va resins. we indi ...201222623327
tonic inhibition of trpv3 by mg2+ in mouse epidermal keratinocytes.the transient receptor potential vanilloid 3 channel (trpv3) is abundantly expressed in epidermal keratinocytes and has important roles in sensory biology and skin health. mg(2+) deficiency causes skin disorders under certain pathological conditions such as type 2 diabetes mellitus. in this study, we investigated the effect of mg(2+) on trpv3 in primary epidermal keratinocytes. extracellular mg(2+) ([mg(2+)](o)) inhibited trpv3-mediated membrane current and calcium influx. trpv3 activation induc ...201222622423
functional dissociation between pikfyve-synthesized ptdins5p and ptdins(3,5)p2 by means of the pikfyve inhibitor ym201636.pikfyve is an essential mammalian lipid kinase with pleiotropic cellular functions whose genetic knockout in mice leads to preimplantation lethality. despite several reports for pikfyve-catalyzed synthesis of phosphatidylinositol 5-phosphate (ptdins5p) along with phosphatidylinositol-3,5-biphosphate [ptdins(3,5)p(2)] in vitro and in vivo, the role of the pikfyve pathway in intracellular ptdins5p production remains underappreciated and the function of the pikfyve-synthesized ptdins5p pool poorly ...201222621786
chemical composition and in vitro cytotoxic activity of essential oil of leaves of malus domestica growing in western himalaya (india).light pale-colored volatile oil was obtained from fresh leaves of malus domestica tree, growing in dhauladhar range of himalaya (himachal pradesh, india), with characteristic eucalyptol dominant fragrance. the oil was found to be a complex mixture of mono-, sesqui-, di-terpenes, phenolics, and aliphatic hydrocarbons. seventeen compounds accounting for nearly 95.3% of the oil were characterized with the help of capillary gc, gc-ms, and nmr. major compounds of the oil were characterized as eucalyp ...201222619691
simultaneous enhancement of cell proliferation and thermally induced harvest efficiency based on temperature-responsive cationic copolymer-grafted microcarriers.the development of large-scale suspension cell cultures using microcarriers has long been a focus of attention in the fields of pharmacy and biotechnology. previously, we developed cell-detachable microcarriers based on temperature-responsive poly(n-isopropylacrylamide) (pipaam)-grafted beads, on which adhering cells can be noninvasively harvested by only reducing the temperature without the need for proteolytic enzyme treatment. in this study, to improve the cell harvest efficiency from bead su ...201222616950
inhibition of forebrain μ-opioid receptor signaling by low concentrations of rimonabant does not require cannabinoid receptors and directly involves μ-opioid receptors.increasing number of publications shows that cannabinoid receptor 1 (cb(1)) specific compounds might act in a cb(1) independent manner, including rimonabant, a potent cb(1) receptor antagonist. opioids, cannabinoids and their receptors are well known for their overlapping pharmacological properties. we have previously reported a prominent decrease in μ-opioid receptor (mor) activity when animals were acutely treated with the putative endocannabinoid noladin ether (ne). in this study, we clarifie ...201222613132
mucin 21 in esophageal squamous epithelia and carcinomas: analysis with glycoform-specific monoclonal antibodies.monoclonal antibodies (mabs) against mucin 21 (muc21), a human counterpart of mouse epiglycanin/muc21, were prepared using human embryonic kidney 293 cells transfected with muc21 as the immunogen. the specificity of these mabs was examined by flow cytometry, immunoprecipitation and western blotting focusing on the differential glycosylation of muc21 expressed in variant chinese hamster ovary (cho) cells (ldld cells and lec2 cells) and cho-k1 cells. one of these mabs, hem21d, bound to both the un ...201222611128
engineering an improved igg4 molecule with reduced disulfide bond heterogeneity and increased fab domain thermal stability.the integrity of antibody structure, stability, and biophysical characterization are becoming increasingly important as antibodies receive increasing scrutiny from regulatory authorities. we altered the disulfide bond arrangement of an igg4 molecule by mutation of the cys at the n terminus of the heavy chain constant domain 1 (c(h)1) (kabat position 127) to a ser and introduction of a cys at a variety of positions (positions 227-230) at the c terminus of c(h)1. an inter-lc-c(h)1 disulfide bond i ...201222610095
analysis of glycosaminoglycan-derived disaccharides by capillary electrophoresis using laser-induced fluorescence detection.a quantitative and highly sensitive method for the analysis of glycosaminoglycan (gag)-derived disaccharides that relies on capillary electrophoresis (ce) with laser-induced fluorescence detection is presented. this method enables complete separation of 17 gag-derived disaccharides in a single run. unsaturated disaccharides were derivatized with 2-aminoacridone to improve sensitivity. the limit of detection was at the attomole level and approximately 100-fold more sensitive than traditional ce-u ...201222609076
down-regulation of the epidermal growth factor receptor by altering n-glycosylation: emerging role of β1,4-galactosyltransferases.blocking n-glycosylation of the epidermal growth factor receptor (egfr) by tunicamycin inhibits its cellular accumulation. due to the toxic potential of this drug, finding less drastic routes to reduce egfr expression is desirable.201222593433
new cell line development for antibody-producing chinese hamster ovary cells using split green fluorescent protein.the establishment of high producer is an important issue in chinese hamster ovary (cho) cell culture considering increased heterogeneity by the random integration of a transfected foreign gene and the altered position of the integrated gene. fluorescence-activated cell sorting (facs)-based cell line development is an efficient strategy for the selection of cho cells in high therapeutic protein production.201222587529
role of the intestinal peptide transporter pept1 in oseltamivir absorption: in vitro and in vivo studies.it was reported that oseltamivir (tamiflu) absorption was mediated by human peptide transporter (hpept) 1. understanding the exact mechanism(s) of absorption is important in the context of drug-drug and diet-drug interactions. hence, we investigated the mechanism governing the intestinal absorption of oseltamivir and its active metabolite (oseltamivir carboxylate) in wild-type [chinese hamster ovary (cho)-k1] and hpept1-transfected cells (cho-pept1), in pharmacokinetic studies in juvenile and ad ...201222584254
oleamide activates peroxisome proliferator-activated receptor gamma (pparγ) in vitro.oleamide (oda) is a fatty acid primary amide first identified in the cerebrospinal fluid of sleep-deprived cats, which exerts effects on vascular and neuronal tissues, with a variety of molecular targets including cannabinoid receptors and gap junctions. it has recently been reported to exert a hypolipidemic effect in hamsters. here, we have investigated the nuclear receptor family of peroxisome proliferator-activated receptors (ppars) as potential targets for oda action.201222584002
the threonine protease activity of testes-specific protease 50 (tsp50) is essential for its function in cell proliferation.testes-specific protease 50 (tsp50), a newly discovered threonine enzyme, has similar amino acid sequences and enzymatic structures to those of many serine proteases. it may be an oncogene. tsp50 is up-regulated in breast cancer epithelial cells, and ectopic expression of tsp50 in tsp50-deficient chinese hamster ovary (cho) cells has been found to promote cell proliferation. however, the mechanisms by which tsp50 exerts its growth-promoting effects are not yet fully understood.201222574111
functional significance of conserved cysteines in the human organic cation transporter 2.the significance of conserved cysteines in the human organic cation transporter 2 (hoct2), namely the six cysteines in the long extracellular loop (loop cysteines) and c474 in transmembrane helix 11, was examined. uptake of tetraethylammonium (tea) and 1-methyl-4-phenypyridinium (mpp) into chinese hamster ovary cells was stimulated >20-fold by hoct2 expression. both cell surface expression and transport activity were reduced considerably following mutation of individual loop cysteines (c51, c63, ...201222573376
znt-1 enhances the activity and surface expression of t-type calcium channels through activation of ras-erk signaling.zinc transporter-1 (znt-1) is a putative zinc transporter that confers cellular resistance from zinc toxicity. in addition, znt-1 has important regulatory functions, including inhibition of l-type calcium channels and activation of raf-1 kinase. here we studied the effects of znt-1 on the expression and function of t-type calcium channels. in xenopus oocytes expressing voltage-gated calcium channel (cav) 3.1 or cav3.2, znt-1 enhanced the low-threshold calcium currents (i(cat)) to 182 ± 15 and 16 ...201222572848
chromatin changes induced by pb and cd in human cells.our earlier data on the apoptotic effects of heavy metals (cd, hg, ni, crvi, and ag) and irradiation (gamma and uv-b) indicated that cellular effects exerted on chromatin structure differ from one other. these observations suggested that chromatin distortions could be characteristic to the genotoxic agent. this notion raised questions whether or not (a) a specific genotoxic agent causes the same chromatin changes in different cell types, (b) closely related toxic compounds could be distinguished ...201222564431
mutagenicity of 5-hydroxymethylfurfural in v79 cells expressing human sult1a1: identification and mass spectrometric quantification of dna adducts formed.5-hydroxymethylfurfural (hmf), a heterocyclic product of the maillard reaction, is a ubiquitous food contaminant. it has demonstrated hepatocarcinogenic activity in female mice. this effect may originate from sulfo conjugation of the benzylic alcohol yielding 5-sulfooxymethylfurfural (smf), which is prone to react with dna via nucleophilic substitution. indeed, we showed that hmf induces gene mutations in chinese hamster v79 cells engineered for the expression of human (h) sulfotransferase (sult ...201222563731
analysis of dynamic changes in the proteome of a bcl-xl overexpressing chinese hamster ovary cell culture during exponential and stationary phases.mammalian cell cultures used for biopharmaceutical production undergo various dynamic biological changes over time, including the transition of cells from an exponential growth phase to a stationary phase during cell culture. to better understand the dynamic aspects of cell culture, a quantitative proteomics approach was used to identify dynamic trends in protein expression over the course of a chinese hamster ovary (cho) cell culture for the production of a recombinant monoclonal antibody and o ...201222556165
real-time detection of acetylcholine release from the human endocrine pancreas.neurons, sensory cells and endocrine cells secrete neurotransmitters and hormones to communicate with other cells and to coordinate organ and system function. validation that a substance is used as an extracellular signaling molecule by a given cell requires a direct demonstration of its secretion. in this protocol we describe the use of biosensor cells to detect neurotransmitter release from endocrine cells in real-time. chinese hamster ovary cells expressing the muscarinic acetylcholine (ach) ...201222555241
enhancing and limiting endothelin-1 signaling with a cell-penetrating peptide mimicking the third intracellular loop of the etb receptor.tat (a 13-mer derived from the hiv-1 tat protein)-linked cell-permeable peptides deliver plasma membrane impermeable cargos into the cell. we investigated the effect of a tat-linked intracellular third loop of the endothelin-1 type b receptor on endothelin-1 activation of erk. the effect of this peptide on erk activation was determined in etb receptor cdna-transfected chinese hamster ovary cells and in eta- and etb-expressing human pulmonary artery smooth muscle cells obtained from a normal and ...201222553998
cell-based microscale isolation of glycoaminoglycans for glycomics study.glycomics research requires the isolation of glycans from cells for structural characterization and functional studies of the glycans. a method for cell-based microscale isolation and quantification of highly sulfated, moderately sulfated, and nonsulfated glycosaminoglycans (gags) was developed using chinese hamster ovary (cho) cells. this microscale isolation relies on a mini-strong anion exchange spin column eluted stepwise with different concentrations of sodium chloride solution. hyaluronic ...201224068855
effects of dapoxetine on cloned kv1.5 channels expressed in cho cells.the effects of dapoxetine were examined on cloned kv1.5 channels stably expressed in chinese hamster ovary cells using the whole-cell patch clamp technique. dapoxetine decreased the peak amplitude of kv1.5 currents and accelerated the decay rate of current inactivation in a concentration-dependent manner with an ic ( 50 ) of 11.6 μm. kinetic analysis of the time-dependent effects of dapoxetine on kv1.5 current decay yielded the apparent association (k (+1 )) and dissociation (k (-1 )) rate const ...201222538641
a thermoprotective role of the sodium channel β1 subunit is lost with the β1 (c121w) mutation.a mutation in the β(1) subunit of the voltage-gated sodium (na(v)) channel, β(1) (c121w), causes genetic epilepsy with febrile seizures plus (gefs+), a pediatric syndrome in which febrile seizures are the predominant phenotype. previous studies of molecular mechanisms underlying neuronal hyperexcitability caused by this mutation were conducted at room temperature. the prevalence of seizures during febrile states in patients with gefs+, however, suggests that the phenotypic consequence of β(1) (c ...201222292491
n-glycans of srec-i (scavenger receptor expressed by endothelial cells): essential role for ligand binding, trafficking and stability.scavenger receptor expressed by endothelial cells (srec-i) mediates the endocytosis of chemically modified lipoproteins such as acetylated low-density lipoprotein (ac-ldl) and oxidized ldl and is implicated in atherogenesis. we produced recombinant srec-i in chinese hamster ovary-k1 cells and identified three potential glycosylation sites, asn(289), asn(382) and asn(393), which were all glycosylated. to determine the function of n-glycans in srec-i, we characterized srec-i mutant proteins by int ...201222279061
5-thiomannosides block the biosynthesis of dolichol-linked oligosaccharides and mimic class i congenital disorders of glycosylation.in a cell-based assay for novel inhibitors, we have discovered that two glycosides of 5-thiomannose, each containing an interglycosidic nitrogen atom, prevented the correct zymogen processing of the prohormone proopiomelanocortinin (pomc) and the transcription factor sterol-regulatory element-binding protein-2 (srebp-2) in mouse pituitary cells and chinese hamster ovary (cho) cells, respectively. in the case of srebp-2, these effects were correlated with the altered n-linked glycosylation of sub ...201222262650
three decades of p-gp inhibitors: skimming through several generations and scaffolds.many tumor cells become resistant to commonly used cytotoxic drugs due to the overexpression of atp-binding cassette (abc) transporters, namely p-glycoprotein (p-gp). the discovery of the reversal of multidrug resistance (mdr) by verapamil occured in 1981, and in 1968 mdr chinese hamster cell lines were isolated for the first time. since then, p-gp inhibitors have been intensively studied as potential mdr reversers. initially, drugs to reverse mdr were not specifically developed for inhibiting p ...201222257057
dna display selection of peptide ligands for a full-length human g protein-coupled receptor on cho-k1 cells.the g protein-coupled receptors (gpcrs), which form the largest group of transmembrane proteins involved in signal transduction, are major targets of currently available drugs. thus, the search for cognate and surrogate peptide ligands for gpcrs is of both basic and therapeutic interest. here we describe the application of an in vitro dna display technology to screening libraries of peptide ligands for full-length gpcrs expressed on whole cells. we used human angiotensin ii (ang ii) type-1 recep ...201222253889
the cytoprotective effect of butin against oxidative stress is mediated by the up-regulation of manganese superoxide dismutase expression through a pi3k/akt/nrf2-dependent pathway.butin (7,3',4'-trihydroxydihydroflavone), a flavonoid with antioxidant activity, was recently reported to protect cells against h2o2-induced apoptosis, oxidative dna damage and oxidative mitochondrial dysfunction. the objective of the present study was to elucidate the mechanism by which butin protects mitochondria. the antioxidant function of manganese superoxide dismutase (mn sod) is important in preventing oxidative stress. while exposure to h2o2 reduced the expression of mn sod in chinese ha ...201222253095
differential in-gel electrophoresis (dige) analysis of cho cells under hyperosmotic pressure: osmoprotective effect of glycine betaine addition.the use of glycine betaine combined with hyperosmolality is known to be an efficient means for achieving high protein production in recombinant chinese hamster ovary (rcho) cells. in order to understand the intracellular events and identify the key factors in rcho cells cultivated with glycine betaine under hyperosmotic conditions, two-dimensional differential in-gel electrophoresis (2d-dige) followed by mass spectrometric analysis was applied. differentially expressed 19 protein spots were sele ...201222252946
probing inhibitory effects of nanocrystalline cellulose: inhibition versus surface charge.ncc derived from different biomass sources was probed for its plausible cytotoxicity by electric cell-substrate impedance sensing (ecis). two different cell lines, spodoptera frugiperda sf9 insect cells and chinese hamster lung fibroblast v79, were exposed to ncc and their spreading and viability were monitored and quantified by ecis. based on the 50%-inhibition concentration (ecis(50)), none of the ncc produced was judged to have any significant cytotoxicity on these two cell lines. however, nc ...201222252333
combined in silico modeling and metabolomics analysis to characterize fed-batch cho cell culture.the increasing demand for recombinant therapeutic proteins highlights the need to constantly improve the efficiency and yield of these biopharmaceutical products from mammalian cells, which is fully achievable only through proper understanding of cellular functioning. towards this end, the current study exploited a combined metabolomics and in silico modeling approach to gain a deeper insight into the cellular mechanisms of chinese hamster ovary (cho) fed-batch cultures. initially, extracellular ...201222252269
cytotoxic purine nucleoside analogues bind to a1, a2a, and a3 adenosine receptors.fludarabine, clofarabine, and cladribine are anticancer agents which are analogues of the purine nucleoside adenosine. these agents have been associated with cardiac and neurological toxicities. because these agents are analogues of adenosine, they may act through adenosine receptors to elicit their toxic effects. the objective of this study was to evaluate the ability of cytotoxic nucleoside analogues to bind and activate adenosine receptor subtypes (a(1), a(2a), a(2b), and a(3)). radioligand b ...201222249336
caveolae optimize tissue factor-factor viia inhibitory activity of cell-surface-associated tissue factor pathway inhibitor.tfpi (tissue factor pathway inhibitor) is an anticoagulant protein that prevents intravascular coagulation through inhibition of fxa (factor xa) and the tf (tissue factor)-fviia (factor viia) complex. localization of tfpi within caveolae enhances its anticoagulant activity. to define further how caveolae contribute to tfpi anticoagulant activity, cho (chinese-hamster ovary) cells were co-transfected with tf and membrane-associated tfpi targeted to either caveolae [tfpi-gpi (tfpi-glycosylphosphat ...201222239091
transgene copy number distribution profiles in recombinant cho cell lines revealed by single cell analyses.clonally derived recombinant cell lines are highly desired to achieve consistent production of recombinant biotherapeutics. despite repeated rounds of cloning by limiting dilution or single cell cloning, the resulting cell lines have often been observed to diverge, becoming a heterogeneous population and losing productivity over long-term sub-culturing. to understand the underlying molecular mechanisms, we developed quantitative polymerase chain reaction (qpcr) assays for the analysis of transge ...201222234778
conversion of tsh heterodimer to a single polypeptide chain increases bioactivity and longevity.tsh is a dimeric glycoprotein hormone composed of a common α-subunit noncovalently linked to a hormone-specific β-subunit. previously, the tsh heterodimer was successfully converted to an active single-chain hormone by genetically fusing α and β genes with [tshβ- carboxyl-terminal peptide (ctp)-α] or without (tshβ-α) the ctp of human chorionic gonadotropin β-subunit as a linker. in the present study, tsh variants were expressed in chinese hamster ovarian cells. the results indicated that tshβ-α ...201222234466
bleomycin induces delayed instability of interstitial telomeric sequences in chinese hamster ovary cells.we analyzed the behavior of interstitial telomeric sequences (itss) in the progeny of chinese hamster ovary (cho) cells exposed to the radiomimetic compound bleomycin (blm) in order to determine if itss play some role in the long-term clastogenic effect of this antibiotic. to this end, cho cells were treated with a single concentration of blm (2.5μg/ml), and the frequency of unstable chromosomal aberrations was determined at several times after treatment (18h, and 6, 15 and 34/36 days) by using ...201222230195
prestin binding peptides as ligands for targeted polymersome mediated drug delivery to outer hair cells in the inner ear.targeted delivery of treatment agents to the inner ear using nanoparticles is an advanced therapeutic approach to cure or alleviate hearing loss. designed to target the outer hair cells of the cochlea, two 12-mer peptides (a(665) and a(666)) with affinity to prestin were identified following 3 rounds of sequential phage display. two-round display with immobilized prestin protein was used to enrich the library for full-length prestin. the last round was performed using cos-7 cells transiently tra ...201222227343
efficient expression of recombinant soluble human fcγri in mammalian cells and its characterization.the extracellular domain of human fcγri which interacts with a human igg was expressed as recombinant soluble human fcγri (rshfcγri) by chinese hamster ovary (cho) cell. stable cho cell clones with efficient expression of rshfcγri were established based on a dihydrofolate reductase (dhfr)/methotrexate (mtx) gene-amplification system. the cho clones efficiently produced rshfcγri under high-density continuous culture in a bioreactor. after 53 days of culture, the number of cells had reached approx ...201222226868
the green tea polyphenol (2)-epigallocatechin-3-gallate (egcg) is not a β-secretase inhibitor.(2)-epigallocatechin-3-gallate (egcg) is a major polyphenolic component of green tea. a number of studies have demonstrated egcg has the possibility for delaying the onset or retarding the progression of alzheimer's disease (ad) and indicated egcg possess inhibition of β-secretase activity. we utilized homogeneous time-resolved fluorescence assay with a substrate eu-cevnldaefk-qsy7 to screen β-secretase inhibitor in a cell-free system and alphalisa assay in cell system. the results first showed ...201222225636
pharmacologic effects of 2-methoxyestradiol on angiotensin type 1 receptor down-regulation in rat liver epithelial and aortic smooth muscle cells.delayed onset of cardiovascular disease (cvd) in female patients is not well understood, but could be due in part to the protective effect of estrogen before menopause. experimental studies have identified the angiotensin type 1 receptor (at1r) as a key factor in the progression of cvd.201222366193
temperature-responsive poly(n-isopropylacrylamide)-grafted microcarriers for large-scale non-invasive harvest of anchorage-dependent cells.cell cultivation on the surface of microcarriers in stirred suspension is an essential method for the large-scale culture of anchorage-dependent cells. for applying this method to the field of cell therapy and for obtaining a large number of intact cells, non-invasive cell harvest without proteolytic enzyme treatment is an advantageous method. in this regard, temperature-responsive microcarriers that bearing poly(n-isopropylacrylamide) (pipaam)-grafted chains on the outermost surface were develo ...201222364728
regulatory mechanisms underlying the modulation of girk1/girk4 heteromeric channels by p2y receptors.the muscarinic k(+) channel (i (k,ach)) is a heterotetramer composed of girk1 (kir3.1) and girk4 (kir3.4) subunits of a g protein-coupled inwardly rectifying channel, and plays an important role in mediating electrical responses to the vagal stimulation in the heart. i (k,ach) displays biphasic changes (activation followed by inhibition) through the stimulation of the purinergic p2y receptors, but the regulatory mechanism involved in these modulation of i (k,ach) by p2y receptors remains to be f ...201222362083
modulating cell-cell communication with a high-throughput label-free cell assay.a high-throughput label-free cell assay for modulating cell-cell communication is demonstrated with the epic® system, a resonant waveguide grating sensor platform. natural killer (nk) cells are known to be able to recognize abnormal cells (e.g., cancer cells and cells presenting intercellular adhesion molecule 1 [icam1] through cell surface receptors) and kill them. in this study, the effect of effecter cells nk92mi on two kinds of target cells, cervical cancer cells (hela) and chinese hamster o ...201222357603
effect of canagliflozin on renal threshold for glucose, glycemia, and body weight in normal and diabetic animal models.canagliflozin is a sodium glucose co-transporter (sglt) 2 inhibitor in clinical development for the treatment of type 2 diabetes mellitus (t2dm).201222355316
identification of the receptors for prolactin-releasing peptide (prrp) and carassius rfamide peptide (c-rfa) in chickens.prolactin-releasing peptide (prrp) and its structurally related peptide, carassius arg-phe-amide peptide (c-rfa), have been reported to play similar roles in regulating food intake and pituitary functions in vertebrates. however, the identity, functionality, and expression of the receptor(s) for prrp and c-rfa remain largely unknown in nonmammalian vertebrates, including birds. in this study, three receptors homologous to mammalian prrp receptor (prrpr), named cprrpr1, cprrpr2, and cc-rfar, resp ...201222355069
at-line nir spectroscopy as effective pat monitoring technique in mab cultivations during process development and manufacturing.the application of at-line nir transmittance spectroscopy on supernatant samples from chinese hamster ovary cells (cho) based monoclonal antibody (mab) cultivation processes spanning several scales from 2.5l to 1000 l, cell-lines and development years is described. the collected and preprocessed spectra were used to do process state estimation and to obtain several culture parameters. multivariate process trajectories were computed from nir spectra acquired at-line. these were used to enhance pr ...201222340110
high frequency of serpina6 polymorphisms that reduce plasma corticosteroid-binding globulin activity in chinese subjects.cortisol is transported by corticosteroid-binding globulin (cbg) in blood. single nucleotide polymorphisms (snp) in the human cbg (serpina6) gene that disrupt cbg production or steroid binding are considered rare.201222337907
effects of high passage cultivation on cho cells: a global analysis.cell lines for industrial pharmaceutical protein production processes need to be robust, fast-growing, and high-producing. in order to find such cells, we performed a high passage cultivation of monoclonal antibody producing chinese hamster ovary (cho) cells in shaking flasks for more than 420 days. examinations of cell growth, productivity, intracellular protein, and metabolite characteristics as well as product transcript and genomic integrate levels revealed substantial differences between su ...201222331235
investigating relationship between transfection and permeabilization by the electric field and/or the pluronic® l64 in vitro and in vivo.electrotransfer can be obtained by the successive delivery of a high voltage short duration pulse (hv) inducing membrane destabilization and then a low voltage long duration pulse (lv), allowing dna electrophoresis (hvlv mode). pluronic® l64 (l64) (fluka, sigma-aldrich, l'isle-d'abeau chesnes, saint-quentin fallavier, france) has permeabilizing properties and amplifies the expression of dna. we aimed to determine whether l64 could have an adjuvant effect on transfection by electrotransfer and wh ...201222328073
metabolic engineering of chinese hamster ovary cells: towards a bioengineered heparin.heparin is the most widely used pharmaceutical to control blood coagulation in modern medicine. a health crisis that took place in 2008 led to a demand for production of heparin from non-animal sources. chinese hamster ovary (cho) cells, commonly used mammalian host cells for production of foreign pharmaceutical proteins in the biopharmaceutical industry, are capable of producing heparan sulfate (hs), a related polysaccharide naturally. since heparin and hs share the same biosynthetic pathway, w ...201222326251
defective activities, but not secretions, resulting from gene point mutations of human mannan-binding lectin.human mannan-binding lectin (mbl) plays a pivotal role in innate immunity. substantial literature supports the belief that three point mutations, cgt52tgt, ggc54gac and gga57gaa, in the collagen-like region (clr) of the human mbl gene, are associated with increased susceptibility to infection, autoimmunity and carcinogenesis. to investigate the mechanisms of mbl deficiency, human wild-type and three variant mbl genes were expressed in cos-7 and chinese hamster ovary (cho) cells. results showed t ...201222323042
geissoschizine methyl ether, an alkaloid in uncaria hook, is a potent serotonin ₁a receptor agonist and candidate for amelioration of aggressiveness and sociality by yokukansan.yokukansan (yks), a traditional japanese medicine, is composed of seven kinds of dried herbs. it is widely prescribed in clinical situation for treating psychiatric disorders such as aggressiveness in patients with dementia. we previously demonstrated that yks and uncaria hook (uh), which is a constituent herb of yks, had a partial agonistic effect to 5-ht(1a) receptors in vitro. however, it has still been unclear whether this in vitro effect is reflected in in vivo, and what the active ingredie ...201222314317
mechanical disruption of mammalian cells in a microfluidic system and its numerical analysis based on computational fluid dynamics.the lysis of mammalian cells is an essential part of different lab-on-a-chip sample preparation methods, which aim at the release, separation, and subsequent analysis of dna, proteins, or metabolites. particularly for the analysis of compartmented in vivo metabolism of mammalian cells, such a method must be very fast compared to the metabolic turnover-rates, it should not affect the native metabolite concentrations, and should ideally leave cell organelles undamaged. so far, no such a method is ...201222311121
structural characterization of full-length nsf and 20s particles.the 20s particle, which is composed of the n-ethylmaleimide-sensitive factor (nsf), soluble nsf attachment proteins (snaps) and the snap receptor (snare) complex, has an essential role in intracellular vesicle fusion events. using single-particle cryo-em and negative stain em, we reconstructed four related three-dimensional structures: chinese hamster nsf hexamer in the atpγs, adp-alfx and adp states, and the 20s particle. these structures reveal a parallel arrangement between the d1 and d2 doma ...201222307055
computational identification of microrna gene loci and precursor microrna sequences in cho cell lines.micrornas (mirnas) have recently entered chinese hamster ovary (cho) cell culture technology, due to their severe impact on the regulation of cellular phenotypes. applications of mirnas that are envisioned range from biomarkers of favorable phenotypes to cell engineering targets. these applications, however, require a profound knowledge of mirna sequences and their genomic organization, which exceeds the currently available information of ~400 conserved mature cho mirna sequences. based on these ...201222306111
toward a new gold standard for early safety: automated temperature-controlled herg test on the patchliner.the patchliner® temperature-controlled automated patch clamp system was evaluated for testing drug effects on potassium currents through human ether-à-go-go related gene (herg) channels expressed in chinese hamster ovary cells at 35-37°c. ic(50) values for a set of reference drugs were compared with those obtained using the conventional voltage clamp technique. the results showed good correlation between the data obtained using automated and conventional electrophysiology. based on these results ...201222303293
genotoxic potential of methyleugenol and selected methyleugenol metabolites in cultured chinese hamster v79 cells.methyleugenol is a substituted alkenylbenzene classified by the european union's scientific committee on food as a genotoxic carcinogen. we addressed cytotoxicity, genotoxicity and mutagenicity caused by methyleugenol and selected oxidative methyleugenol metabolites in chinese hamster lung fibroblasts v79 cells. cytotoxicity was measured by two cell proliferation assays, water soluble tetrazolium salt (wst) 1 and sulforhodamine b (srb) assays. genotoxicity was determined by using single cell gel ...201222302122
apolipoprotein a-i amyloidogenic variant l174s, expressed and isolated from stably transfected mammalian cells, is associated with fatty acids.sixteen variants of apolipoprotein a-i (apoa-i) are associated with hereditary systemic amyloidoses, characterized by amyloid deposition in peripheral organs of patients. as these are heterozygous for the amyloidogenic variants, their isolation from plasma is impracticable and recombinant expression systems are needed. here we report the expression of recombinant apoa-i amyloidogenic variant leu174 with ser (l174s) in stably transfected chinese hamster ovary-k1 cells. apoa-i variant l174s was fo ...201222295944
influence of the accessory protein set on m3 muscarinic receptor phosphorylation and g protein coupling.the proto-oncogene and inhibitor of protein phosphatase 2a (pp2a), set, interacts with the third intracellular loop of the m3 muscarinic receptor (m3-mr), and set knockdown with small interfering rna (sirna) in chinese hamster ovary (cho) cells augments m3-mr signaling. however, the mechanism of this action of set on receptor signaling has not been defined, and we initiated studies to address this question. knockdown of set by sirna in cho cells stably expressing the m3-mr did not alter agonist- ...201222466417
engulfment protein gulp is regulator of transforming growth factor-β response in ovarian cells.transforming growth factor β (tgf-β) is a key regulatory molecule with pleiotropic effects on cell growth, migration, and invasion. as a result, impairment of proper tgf-β signaling is central to tumorigenesis and metastasis. the tgf-β receptor v (tgfbrv or lrp1) has been shown to be responsible for tgf-β-mediated cell growth inhibition in chinese hamster ovary (cho) cells. the lrp1 adapter protein gulp mediates internalization of the various lrp1-specific ligands, and we hypothesize that gulp a ...201222451657
advanced microscale bioreactor system: a representative scale-down model for bench-top bioreactors.in recent years, several automated scale-down bioreactor systems have been developed to increase efficiency in cell culture process development. ambr™ is an automated workstation that provides individual monitoring and control of culture dissolved oxygen and ph in single-use, stirred-tank bioreactors at a working volume of 10-15 ml. to evaluate the ambr™ system, we compared the performance of four recombinant chinese hamster ovary cell lines in a fed-batch process in parallel ambr™, 2-l bench-to ...201222451076
cytotoxicity and genotoxicity evaluation of antidote oxime hi-6 tested on eight cell lines of human and rodent origin.oxime hi-6 is an efficient reactivator of the acetylcholinesterase inhibited by organophosphorous nerve agents. in this study we have estimated cytotoxicity of hi-6 by the colony forming assay and genotoxicity by the comet assay on human and rodent cell lines. ic50 of hi-6 assessed by the colony forming capacity was 3.59 mm for hela cells and 5.18 mm for a mouse cell line l929. small difference in cytotoxicity was found among other cell lines tested: ic50 was 1.61 mm for human a549 cells, 1.14 m ...201222447833
impact of mir-7 over-expression on the proteome of chinese hamster ovary cells.micrornas play critical roles in the regulation of biological processes such as growth, apoptosis, productivity and secretion thus representing a potential route toward enhancing desirable characteristics of mammalian cells for biopharmaceutical production. we have previously found that mir-7 over-expression significantly inhibits the growth of cho-seap cells without impacting cellular viability, with an associated increase in normalised productivity. understanding the biological basis of this e ...201222445466
cell type-specific β2-adrenergic receptor clusters identified using photoactivated localization microscopy are not lipid raft related, but depend on actin cytoskeleton integrity.recent developments in the field of optical super-resolution techniques allow both a 10-fold increase in resolution as well as an increased ability to quantify the number of labeled molecules visualized in the fluorescence measurement. by using photoactivated localization microscopy (palm) and an experimental approach based on the systematic comparison with a nonclustering peptide as a negative control, we found that the prototypical g protein-coupled receptor β2-adrenergic receptor is partially ...201222442147
fibrinogen residue γala341 is necessary for calcium binding and 'a-a' interactions.the fibrinogen γ-module has several important sites relating to fibrinogen function, which include the high affinity calcium binding site, hole 'a' that binds with knob 'a', and the d:d interface. residue γala341, which is located in the vicinity of these sites, is altered in three variant fibrinogens: fibrinogen seoul (γala341asp), tolaga bay (γala341val), and lyon iii (γala341thr). in order to investigate the impaired polymerisation of fibrinogens γala341asp and γala341val to understand the ro ...201222437918
characterisation of a setup for mixed beam exposures of cells to 241am alpha particles and x-rays.exposure of humans to mixed fields of high- and low-linear energy transfer (let) radiation occurs in many situations-for example, in urban areas with high levels of indoor radon as well as background gamma radiation, during airplane flights or certain forms of radiation therapy. from the perspective of health risk associated with exposure to mixed fields, it is important to understand the interactions between different radiation types. in most cellular investigations on mixed beams, two types of ...201222434924
acute and subacute toxicity and genotoxicity of schizonepetin, a naturally occurring monoterpene with antiviral activity.in the present study, the acute, subacute and genetic toxicity of schizonepetin was assessed. the median lethal dose (ld(50)) of schizonepetin after oral administration was 478 mg/kg body weight in mice. studies on dose toxicity were repeatedly conducted at 0, 60, 120, and 240 mg/kg bw/day in rats for 35 days after oral administration. based on the results of this study, a dose level of 120 mg/kg bw/day is considered the no-observed-adverse-effect-level (noael) in rats. schizonepetin was negativ ...201222429819
secondary metabolites, cytotoxic response by neutral red retention and protective effect against h2o2 induced cytotoxicity of sedum caespitosum.the etoac, n-buoh and h20 subextracts of the crude meoh extract of the aerial parts of sedum caespitosum (cav.) dc. were screened for cytotoxicity using the neutral red assay in chinese hamster ovary cells as well as their protective effect against h2o2 induced cytotoxicity in human red blood cells. while the extracts did not show cytotoxicity, they displayed a protective effect compared to a blank and ascorbic acid. gallic acid (1), kaempferol 3-o-alpha-rhamnopyranoside (2), quercetin 3-o-beta- ...201222428239
role of non-specific dna in reducing coding dna requirement for transient gene expression with cho and hek-293e cells.transient gene expression (tge) is a rapid method for the production of recombinant proteins in mammalian cells. while the tge volumetric productivity has improved significantly over the past decade, the amount of plasmid dna (pdna) needed for transfection remains very high. here, we examined the use of non-specific (filler) dna to partially replace the transgene-bearing plasmid dna (coding pdna) in transfections of chinese hamster ovary (cho) and human embryo kidney (hek-293e) cells. when the o ...201222422519
conditioned medium from actinomycin d-treated apoptotic cells induces mitochondria-dependent apoptosis in bystander cells.chemical-induced bystander effects have been known for several years, but the underlying mechanism is still seldom investigated. previous researchers have found that mitomycin c and phleomycin induced micronuclei in bystander cells the same as in exposed cells. we previously demonstrated the ability of actinomycin d (actd) to induce bystander effects in normal chinese hamster fibroblast v79 cells and found that conditioned medium (cm) obtained from actd-exposed apoptotic cells induced apoptosis ...201222421271
molecular determinants of ligand selectivity for the human multidrug and toxin extruder proteins mate1 and mate2-k.the present study compared the selectivity of two homologous transport proteins, multidrug and toxin extruders 1 and 2-k (mate1 and mate2-k), and developed three-dimensional pharmacophores for inhibitory ligand interaction with human mate1 (hmate1). the human orthologs of mate1 and mate2-k were stably expressed in chinese hamster ovary cells, and transport function was determined by measuring uptake of the prototypic organic cation (oc) substrate 1-methyl-4-phenylpyridinium (mpp). both mates had ...201222419765
characterization of glucagon-like peptide 1 receptor (glp1r) gene in chickens: functional analysis, tissue distribution, and identification of its transcript variants.glucagon-like peptide 1 (glp1) receptor plays a critical role in mediating the biological actions of glp1 in mammals and fish; however, the gene structure, expression, and functionality of glp1 receptor (glp1r) remain largely unknown in birds. in this study, the full-length cdna of chicken glp1r (cglp1r) was first cloned from brain tissue by reverse transcription pcr. the putative cglp1r is 459 amino acids in length and shares high amino acid sequence identity with that of human (79%), rat (80%) ...201222417644
cytotoxicity and genotoxicity of root canal sealers based on mineral trioxide aggregate.mta has good biological properties, and it is a mineralization-inducing material with different indications in endodontics. initially this material was not recommended as root canal sealer. however, a resin sealer based on mineral trioxide aggregate (mta fillapex) was recently released with this indication. because mta is in contact with the periodontal tissues, bone, and pulp, it is important to know its cytotoxic and genotoxic effects. the purpose of this study was to evaluate the cytotoxicity ...201222414836
in vitro and in vivo evaluation of the genotoxicity of gumiganghwal-tang, a traditional herbal prescription.gumiganghwal-tang (ggt, known as kumi-kyokatsu-to in japanese) is a traditional herbal prescription made from nine different herbs that is used for the treatment of the common cold, pain, and inflammatory diseases.201222414481
regulation of gip and glp1 receptor cell surface expression by n-glycosylation and receptor heteromerization.in response to a meal, glucose-dependent insulinotropic polypeptide (gip) and glucagon-like peptide-1 (glp-1) are released from gut endocrine cells into the circulation and interact with their cognate g-protein coupled receptors (gpcrs). receptor activation results in tissue-selective pleiotropic responses that include augmentation of glucose-induced insulin secretion from pancreatic beta cells. n-glycosylation and receptor oligomerization are co-translational processes that are thought to regul ...201222412906
a screening method to assess biological effects of microrna overexpression in chinese hamster ovary cells.micrornas (mirnas) are a novel class of short non-coding rnas, which negatively regulate target gene expression at post-transcriptional level. they mediate an important layer of control in the global regulation of gene networks, controlling a broad range of physiological as well as patho-physiological pathways including development, cancer, metabolism, proliferation, and stress resistance. so far, more than 365 mirna genes have been identified in cho cells. the functional analysis of the physiol ...201222407745
compromising σ-1 receptors at the endoplasmic reticulum render cytotoxicity to physiologically relevant concentrations of dopamine in a nuclear factor-κb/bcl-2-dependent mechanism: potential relevance to parkinson's disease.the endoplasmic reticulum (er) chaperone σ-1 receptor (sig-1r) is cytoprotective against er stress-induced apoptosis. the level of sig-1rs in the brain was reported to be lower in early parkinsonian patients. because dopamine (da) toxicity is well known to be involved in the etiology of parkinson's disease, we tested in this study whether a relationship might exist between sig-1rs and da-induced cytotoxicity in a cellular model by using chinese hamster ovary (cho) cells. da in physiological conc ...201222399814
association of dna double-strand break gene xrcc6 genotypes and lung cancer in taiwan.the dna repair gene x-ray repair complementing defective repair in chinese hamster cells 6 (xrcc6) is thought to play an important role in the repair of dna double-strand breaks. it is known that defects in double-strand break repair capacity can lead to irreversible genomic instability. however, the association of polymorphic variants of xrcc6 with lung cancer susceptibility has never been reported. in this hospital-based case-control study, the association of xrcc6 promoter t-991c (rs5751129), ...201222399625
imb2026791, a xanthone, stimulates cholesterol efflux by increasing the binding of apolipoprotein a-i to atp-binding cassette transporter a1.it is known that the atp-binding cassette transporter a1 (abca1) plays a major role in cholesterol homeostasis and high density lipoprotein (hdl) metabolism. several laboratories have demonstrated that abca1 binding to lipid-poor apolipoprotein a-i (apoa-i) will mediate the assembly of nascent hdl and cellular cholesterol efflux, which suggests a possible receptor-ligand interaction between abca1 and apoa-i. in this study, a cell-based-elisa-like high-throughput screening (hts) method was develo ...201222399138
testosterone synthesis in patients with 17β-hydroxysteroid dehydrogenase 3 deficiency.17β-hydroxysteroid dehydrogenase 3 (17β-hsd 3) deficiency is a rare cause of 46,xy disorders of sex development (dsd). at puberty, these patients experience a surge of androstenedione and also testosterone, leading to substantial virilization. the origin of testosterone synthesis in these patients remains elusive. we investigated the expression of the isoenzyme akr1c3 (17β-hsd 5) in the testis and patient-derived genital skin fibroblasts (gsf) as well as the ability of gsf to synthesize testoste ...201222398356
hspa2 overexpression protects v79 fibroblasts against bortezomib-induced apoptosis.human hspa2 is a member of the hspa (hsp70) family of heat-shock proteins, encoded by the gene originally described as testis-specific. recently, it has been reported that hspa2 can be also expressed in human somatic tissues in a cell-type specific manner. the aim of the present study was to find out whether hspa2 can increase the resistance of somatic cells to the toxic effect of heat shock, proteasome inhibitors, and several anticancer cytostatics. we used a chinese hamster fibroblast v79 cell ...201222397456
utilization and evaluation of cho-specific sequence databases for mass spectrometry based proteomics.recently released sequence information on chinese hamster ovary (cho) cells promises to not only facilitate our understanding of these industrially important cell factories through direct analysis of the sequence, but also to enhance existing methodologies and allow new tools to be developed. in this article we demonstrate the utilization of cho specific sequence information to improve mass spectrometry (ms) based proteomic identification. the use of various cho specific databases enabled the id ...201222389098
multiple targeting motifs direct nramp1 into lysosomes.natural resistance-associated macrophage protein 1 (nramp1) containing 548 amino acids (aa) and 12 transmembrane domains (tmds) is localized in membranes of lysosomes. our study aimed to investigate the targeting motifs of nramp1 by expressing gfp-tagged full-length and truncated nramp1 proteins and overlapping with the lysosomal marker lamp1-rfp in chinese hamster ovary (cho) cells. the nh(2)-terminal amino acids 73-140 region including tmd2 was essential for nramp1 lysosomal targeting. the aa. ...201222382021
two-photon fluorescence microscopy imaging of cellular oxidative stress using profluorescent nitroxides.a range of varying chromophore nitroxide free radicals and their nonradical methoxyamine analogues were synthesized and their linear photophysical properties examined. the presence of the proximate free radical masks the chromophore's usual fluorescence emission, and these species are described as profluorescent. two nitroxides incorporating anthracene and fluorescein chromophores (compounds 7 and 19, respectively) exhibited two-photon absorption (2pa) cross sections of approximately 400 g.m. wh ...201222380794
genomic instability induced by α-pinene in chinese hamster cell line.here, we report the effects of exposure of mammalian cells to α-pinene, a bicyclic monoterpene used in insecticides, solvents and perfumes. morphological analysis, performed in v79-cl3 cells exposed for 1 h to increasing concentrations (25 up to 50 μm) of α-pinene, indicated a statistically significant increase in micronucleated and multinucleated cell frequencies; apoptotic cells were seen at 40 and 50 μm. this monoterpene caused genomic instability by interfering with mitotic process; in fact, ...201222379123
synthetic lethal targeting of dna double-strand break repair deficient cells by human apurinic/apyrimidinic endonuclease inhibitors.an apurinic/apyrimidinic (ap) site is an obligatory cytotoxic intermediate in dna base excision repair (ber) that is processed by human ap endonuclease 1 (ape1). ape1 is essential for ber and an emerging drug target in cancer. we have isolated novel small molecule inhibitors of ape1. in this study, we have investigated the ability of ape1 inhibitors to induce synthetic lethality (sl) in a panel of dna double-strand break (dsb) repair deficient and proficient cells; i) chinese hamster (ch) cells: ...201222377908
adjustment of ultrasound exposure duration to microbubble sonodestruction kinetics for optimal cell sonoporation in vitro.cell sonoporation enables the delivery of various exogenous molecules into the cells. to maximize the percentage of reversibly sonoporated cells and to increase cell viability we propose a model for implicit dosimetry for adjustment of ultrasound (us) exposure duration. the chinese hamster ovary cell suspension was supplemented with microbubbles (mb) and exposed to us, operating at the frequency of 880khz, with a 100% duty cycle and with an output peak negative pressure (pnp) of 500kpa for durat ...201222376133
Refresh
firstprev
...31323334353637383940...
nextlast
Displaying items 3601 - 3700 of 26566