Publications

mathematical models describing chinese hamster ovary cell death due to electroporation in vitro.	electroporation is a phenomenon used in the treatment of tumors by electrochemotherapy, non-thermal ablation with irreversible electroporation, and gene therapy. when treating patients, either predefined or variable electrode geometry is used. optimal pulse parameters are predetermined for predefined electrode geometry, while they must be calculated for each specific case for variable electrode geometry. the position and number of electrodes are also determined for each patient. it is currently ...	2015	26223863
re-programming cho cell metabolism using mir-23 tips the balance towards a highly productive phenotype.	microrna engineering of cho cells has already proved successful in enhancing various industrially relevant phenotypes and producing various recombinant products. a single mirna's ability to interact with multiple mrna targets allows their regulatory capacity to extend to processes such as cellular metabolism. various metabolic states have previously been associated with particular cho cell phenotypes such as glycolytic or oxidative metabolism accommodating growth and productivity, respectively. ...	2015	26097147
cy5 total protein normalization in western blot analysis.	western blotting is a widely used method for analyzing specific target proteins in complex protein samples. housekeeping proteins are often used for normalization to correct for uneven sample loads, but these require careful validation since expression levels may vary with cell type and treatment. we present a new, more reliable method for normalization using cy5-prelabeled total protein as a loading control. we used a prelabeling protocol based on cy5 n-hydroxysuccinimide ester labeling that pr ...	2015	26095394
induction of cytotoxic and genotoxic responses by natural and novel quercetin glycosides.	the flavonoids quercetin, and its natural glycosides isoquercetin and rutin, are phytochemicals commonly consumed in plant-derived foods. semi-synthetic water-soluble isoquercetin and rutin glycosides, maltooligosyl isoquercetin, monoglucosyl rutin and maltooligosyl rutin were developed by synthetic glycosylation to overcome solubility challenges for improved incorporation in food and medicinal applications. quercetin and its natural glycosides are known to induce genetic instability and decreas ...	2015	26046972
engineering mammalian cell factories with sineup noncoding rnas to improve translation of secreted proteins.	whenever the function of a recombinant protein depends on post-translational processing, mammalian cells become an indispensable tool for their production. this is particularly true for biologics and therapeutic monoclonal antibodies (mabs). despite some drawbacks, chinese hamster ovary (cho) cells are the workhorse for mabs production in academia and industry. several methodologies have been adopted to improve expression and stability, including methods based on selective pressure or cell engin ...	2015	26045368
fully human her2/cluster of differentiation 3 bispecific antibody triggers potent and specific cytotoxicity of t lymphocytes against breast cancer.	the use of a bispecific antibody (bsab) is a promising and highly specific approach to cancer therapy. in the present study, a fully human recombinant single chain variable fragment bsab against human epidermal growth factor receptor (her)2 and cluster of differentiation (cd)3 was constructed with the aim of developing an effective treatment for breast cancer. her2/cd3 bsab was expressed in chinese hamster ovary cells and purified via nickel column chromatography. flow cytometry revealed that th ...	2015	25760691
rapid recombinant protein production from piggybac transposon-mediated stable cho cell pools.	heterogeneous populations of stably transfected cells (cell pools) can serve for the rapid production of moderate amounts of recombinant proteins. here, we propose the use of the piggybac (pb) transposon system to improve the productivity and long-term stability of cell pools derived from chinese hamster ovary (cho) cells. pb is a naturally occurring genetic element that has been engineered to facilitate the integration of a transgene into the genome of the host cell. in this report pb-derived c ...	2015	25758242
purification of monoclonal antibodies from clarified cell culture fluid using protein a capture continuous countercurrent tangential chromatography.	recent studies using simple model systems have demonstrated that continuous countercurrent tangential chromatography (cctc) has the potential to overcome many of the limitations of conventional protein a chromatography using packed columns. the objective of this work was to optimize and implement a cctc system for monoclonal antibody purification from clarified chinese hamster ovary (cho) cell culture fluid using a commercial protein a resin. several improvements were introduced to the previous ...	2015	25747172
comparative studies of three 68ga-labeled [des-arg10]kallidin derivatives for imaging bradykinin b1 receptor expression with pet.	bradykinin b1 receptor (b1r) is a g-protein-coupled receptor that is overexpressed in a variety of cancers. b1r is not expressed in healthy tissues, making it an attractive cancer imaging marker. previously, we reported selective uptake of (68)ga-p03034 ((68)ga-dota-dpeg2-lys-arg-pro-hyp-gly-cha-ser-pro-leu) in b1r-positive (b1r+) hek293t::hb1r tumor xenografts in mice. in this study, we compare (68)ga-p03034 with (68)ga-labeled p04158 ((68)ga-dota-dpeg2-lys-lys-arg-pro-hyp-gly-igl-ser-d-igl-oic ...	2015	25745086
generation of specific monoclonal antibodies against the extracellular loops of human claudin-3 by immunizing mice with target-expressing cells.	human claudin-3 (cldn3) is a tetraspanin transmembrane protein of tight junction structures and is known to be over-expressed in some malignant tumors. although a specific monoclonal antibody (mab) against the extracellular domains of cldn3 would be a valuable tool, generation of such mabs has been regarded as difficult using traditional hybridoma techniques, because of the conserved sequence homology of cldn3s among various species. in addition, high sequence similarity is shared among claudin ...	2015	25744656
igg aggregation mechanism for cho cell lines expressing excess heavy chains.	aggregates in protein therapeutics like igg monoclonal antibodies (mab) are detrimental to product safety and efficacy. it has been reported that aggregates form in chinese hamster ovary (cho) cell lines expressing greater amount of heavy chain (hc) than light chain (lc). in this study, we observed that aggregates could form within the cells with excess hc and were partially secreted into the supernatant. the aggregates in the supernatant consisted of mainly hc and were partially dissociated und ...	2015	25744439
heat shock protein 27 overexpression in cho cells modulates apoptosis pathways and delays activation of caspases to improve recombinant monoclonal antibody titre in fed-batch bioreactors.	cho cells are major production hosts for recombinant biologics including the rapidly expanding recombinant monoclonal antibodies (mabs). heat shock protein 27 (hsp27) expression was observed to be down-regulated towards the late-exponential and stationary phase of cho fed-batch bioreactor cultures, whereas hsp27 was found to be highly expressed in human pathological cells and reported to have anti-apoptotic functions. these phenotypes suggest that overexpression of hsp27 is a potential cell line ...	2015	25740626
the major dna repair pathway after both proton and carbon-ion radiation is nhej, but the hr pathway is more relevant in carbon ions.	the purpose of this study was to identify the roles of non-homologous end-joining (nhej) or homologous recombination (hr) pathways in repairing dna double-strand breaks (dsbs) induced by exposure to high-energy protons and carbon ions (c ions) versus gamma rays in chinese hamster cells. two chinese hamster cell lines, ovary aa8 and lung fibroblast v79, as well as various mutant sublines lacking dna-pkcs (v3), x-ray repair cross-complementing protein-4 [xrcc4 (xr1), xrcc3 (irs1sf) and xrcc2 (irs1 ...	2015	25738894
cell type-specific response to high intracellular loading of polyacrylic acid-coated magnetic nanoparticles.	magnetic nanoparticles (nps) are a special type of np with a ferromagnetic, electron-dense core that enables several applications such as cell tracking, hyperthermia, and magnetic separation, as well as multimodality. so far, superparamagnetic iron oxide nps (spions) are the only clinically approved type of metal oxide nps, but cobalt ferrite nps have properties suitable for biomedical applications as well. in this study, we analyzed the cellular responses to magnetic cobalt ferrite nps coated w ...	2015	25733835
understanding of altered n-glycosylation-related gene expression in recombinant chinese hamster ovary cells subjected to elevated ammonium concentration by digital mrna counting.	to understand the effects of ammonium on n-glycosylation, recombinant chinese hamster ovary (rcho) cells that produce the fc-fusion protein were cultivated in serum-free suspension cultures with 10 mm ammonium addition. the addition of ammonium to the cultures reduced the relative proportion of acidic isoforms and sialic acid content of an fc-fusion protein. fifty two n-glycosylation-related gene expressions were assessed by the nanostring ncounter system, which provides a digital readout using ...	2015	25728222
overproduction of recombinant human transforming growth factor beta 3 in chinese hamster ovary cells.	transforming growth factor beta 3 (tgfβ3) is an important cytokine, functioning in cell proliferation and differentiation, and has been considered to have therapeutic potential for treating various diseases and for scar reduction in adult wound healing. in the current study, a chinese hamster ovary (cho) cell line overexpressing recombinant human tgfβ3 (rhtgfβ3) was established. through a 15-day fed-batch culture process in a 7.5-l bioreactor (5-l working volume) using chemically defined medium, ...	2015	25727048
genotoxicity assessment of melamine in the in vivo pig-a mutation assay and in a standard battery of assays.	the genotoxicity of melamine was evaluated with the combined pig-a mutation/micronucleus assay, the bacterial reverse mutation assay, and the in vitro cytokinesis-block micronucleus assay (cbmn). five groups of six- to eight-week-old male sprague-dawley (sd) rats were given three daily doses of vehicle control (100% pure sesame oil), melamine (500, 1000, and 2000 mg/kg) or positive control (n-ethyl-n-nitrosourea, enu, 20 mg/kg) by oral gavage. peripheral blood was sampled pre-dose (day -1) and a ...	2015	25726176
triphenyl phosphate-induced developmental toxicity in zebrafish: potential role of the retinoic acid receptor.	using zebrafish as a model, we previously reported that developmental exposure to triphenyl phosphate (tpp) - a high-production volume organophosphate-based flame retardant - results in dioxin-like cardiac looping impairments that are independent of the aryl hydrocarbon receptor. using a pharmacologic approach, the objective of this study was to investigate the potential role of retinoic acid receptor (rar) - a nuclear receptor that regulates vertebrate heart morphogenesis - in mediating tpp-ind ...	2015	25725299
role of isolated and clustered dna damage and the post-irradiating repair process in the effects of heavy ion beam irradiation.	clustered dna damage is a specific type of dna damage induced by ionizing radiation. any type of ionizing radiation traverses the target dna molecule as a beam, inducing damage along its track. our previous study showed that clustered dna damage yields decreased with increased linear energy transfer (let), leading us to investigate the importance of clustered dna damage in the biological effects of heavy ion beam radiation. in this study, we analyzed the yield of clustered base damage (comprisin ...	2015	25717060
regulation of blood pressure, oxidative stress and at1r by high salt diet in mutant human dopamine d5 receptor transgenic mice.	humans have dopamine d5 receptors (hd5r) with single-nucleotide polymorphisms and a diminished function. we generated hd5(f173l) cdna that has a decreased response to d5r agonist-mediated increase in camp production and increased production of reactive oxygen species, relative to wild-type hd5r (hd5(wt)) cdna expressed in chinese hamster ovary cells. to investigate the role of hd5(f173l) in the pathogenesis of salt-sensitive hypertension, we generated transgenic mice overexpressing hd5(f173l) or ...	2015	25716648
a consensus approach for estimating the predictive accuracy of dynamic models in biology.	mathematical models that predict the complex dynamic behaviour of cellular networks are fundamental in systems biology, and provide an important basis for biomedical and biotechnological applications. however, obtaining reliable predictions from large-scale dynamic models is commonly a challenging task due to lack of identifiability. the present work addresses this challenge by presenting a methodology for obtaining high-confidence predictions from dynamic models using time-series data. first, t ...	2015	25716416
site-specific integration in cho cells mediated by crispr/cas9 and homology-directed dna repair pathway.	chinese hamster ovary (cho) cells are the most widely used mammalian hosts for production of therapeutic proteins. however, development of recombinant cho cell lines has been hampered by unstable and variable transgene expression caused by random integration. here we demonstrate efficient targeted gene integration into site-specific loci in cho cells using crispr/cas9 genome editing system and compatible donor plasmid harboring a gene of interest (goi) and short homology arms. this strategy has ...	2015	25712033
pdadmac flocculation of chinese hamster ovary cells: enabling a centrifuge-less harvest process for monoclonal antibodies.	high titer (>10 g/l) monoclonal antibody (mab) cell culture processes are typically achieved by maintaining high viable cell densities over longer culture durations. a corresponding increase in the solids and sub-micron cellular debris particle levels are also observed. this higher burden of solids (≥15%) and sub-micron particles typically exceeds the capabilities of a continuous centrifuge to effectively remove the solids without a substantial loss of product and/or the capacity of the harvest ...	2015	25706650
phosphatidylinositol4-phosphate 5-kinase prevents the decrease in the herg potassium current induced by gq protein-coupled receptor stimulation.	the human ether-a-go-go-related gene (herg) potassium current (iherg) has been shown to decrease in amplitude following stimulation with gq protein-coupled receptors (gqrs), such as α1-adrenergic and m1-muscarinic receptors (α1r and m1r, respectively), at least partly via the reduction of membrane phosphatidylinositol 4,5-bisphosphate (pi(4,5)p2). the present study was designed to investigate the modulation of herg channels by pi(4,5)p2 and phosphatidylinositol4-phosphate 5-kinase (pi(4)p5-k), a ...	2015	25704028
the dna methylation landscape of chinese hamster ovary (cho) dp-12 cells.	chinese hamster ovary (cho) cells represent the most commonly used production cell line for therapeutic proteins. by recent genome and transcriptome sequencing a basis was created for future investigations of genotype-phenotype relationships and for improvement of cho cell productivity and product quality. in this context information is missing about dna cytosine methylation as a crucial epigenetic modification and an important element in mammalian genome regulation and development. here, we pre ...	2015	25701679
characterization of a recombinant humanized anti-cocaine monoclonal antibody and its fab fragment.	variations of post-translational modifications are important for stability and in vivo behavior of therapeutic antibodies. a recombinant humanized anti-cocaine monoclonal antibody (h2e2) was characterized for heterogeneity of n-linked glycosylation and disulfide bonds. in addition, charge heterogeneity, which is partially due to the presence or absence of c-terminal lysine on the heavy chains, was examined. for cocaine overdose therapy, fab fragments may be therapeutic, and thus, a simplified me ...	2015	25692880
annotation of additional evolutionary conserved micrornas in cho cells from updated genomic data.	micrornas are small non-coding rnas that play a critical role in post-transcriptional control of gene expression. recent publications of genomic sequencing data from the chinese hamster (cgr) and chinese hamster ovary (cho) cells provide new tools for the discovery of novel mirnas in this important production system. version 20 of the mirna registry mirbase contains 307 mature mirnas and 200 precursor sequences for cgr/cho. we searched for evolutionary conserved mirnas from mirbase v20 in recent ...	2015	25689160
transcriptional and post-transcriptional limitations of high-yielding, pei-mediated transient transfection with cho and hek-293e cells.	transient gene expression (tge) in human embryonic kidney (hek-293) and chinese hamster ovary (cho) cells is a well-established technology for the rapid generation of recombinant proteins. although the maximum tge yields have reached 1 g/l or more, the amount of plasmid dna (pdna) required for transfection remains high. although greater than 10(3) copies of pdna are present per transfected cell, protein yields are still lower than those achieved in recombinant cell lines with only one or a few c ...	2015	25683738
efficient enrichment of high-producing recombinant chinese hamster ovary cells for monoclonal antibody by flow cytometry.	to screen a high-producing recombinant chinese hamster ovary (cho) cell from transfected cells is generally laborious and time-consuming. we developed an efficient enrichment strategy for high-producing cell screening using flow cytometry (fcm). a stable pool that had possibly shown a huge variety of monoclonal antibody (mab) expression levels was prepared by transfection of an expression vector for mab production to a cho cell. to enrich high-producing cells derived from a stable pool stained w ...	2015	25683450
development and characterisation of novel fentanyl-delta opioid receptor antagonist based bivalent ligands.	opioid tolerance is a limiting factor in chronic pain. delta opioid peptide (dop)(δ) receptor antagonism has been shown to reduce tolerance. here, the common clinical mu opioid peptide (mop)(µ) receptor agonist fentanyl has been linked to the dop antagonist dmt-tic (2',6'-dimethyl-l-tyrosyl-1,2,3,4-tetrahydrisoquinoline-3-carboxylic acid) to create new bivalent compounds.	2015	25680364
optimization of chemically defined feed media for monoclonal antibody production in chinese hamster ovary cells.	chinese hamster ovary (cho) cells are the most commonly used mammalian host for large-scale commercial production of therapeutic monoclonal antibodies (mabs). chemically defined media are currently used for cho cell-based mab production. an adequate supply of nutrients, especially specific amino acids, is required for cell growth and mab production, and chemically defined fed-batch processes that support rapid cell growth, high cell density, and high levels of mab production is still challenging ...	2015	25678240
identification of a functional antioxidant responsive element in the promoter of the chinese hamster carbonyl reductase 3 (chcr3) gene.	chcr3, a member of the short-chain dehydrogenase/reductase superfamily, is a carbonyl reductase 3 enzyme in chinese hamsters. carbonyl reductase 3 in humans has been believed to involve the metabolism and/or pharmacokinetics of anthracycline drugs, and the mechanism underlying the gene regulation has been investigated. in this study, the nucleotide sequence of the chcr3 promoter was originally determined, and its promoter activity was characterised. the proximal promoter region is tata-less and ...	2015	25677373
codon optimization of genes for efficient protein expression in mammalian cells by selection of only preferred human codons.	a simple design method for codon optimization of genes to express a heterologous protein in mammalian cells is described. codon optimization was performed by choosing only codons preferentially used in humans and with over 60% gc content, and the method was named the "preferred human codon-optimized method." to test our simple rule for codon optimization, the preferred human codon-optimized genes for six proteins containing photoproteins (aequorin and clytin ii) and luciferases (gaussia lucifera ...	2015	25665506
nonacog gamma, a novel recombinant factor ix with low factor ixa content for treatment and prophylaxis of bleeding episodes.	nonacog gamma is a new recombinant factor ix to treat factor ix deficiency. it is indicated for control of bleeding episodes, perioperative management and routine prophylaxis to prevent or reduce the frequency of bleeding episodes in adults and children with hemophilia b. nonacog gamma was first approved in the usa in june 2013 under the trade name rixubis followed by market approvals in australia and the eu in 2014, and marketing authorization decision is pending in japan. nonacog gamma is deri ...	2015	25660348
in vitro evaluation of biocompatibility of ti-mo-sn-zr superelastic alloy.	shape memory alloys (smas) including superelastic alloys have unique properties such as shape memory and superelasticity, thus they are recognized as very useful biomaterials. these properties are very advantageous for medical use, and actually the sma wires have been widely used in medical field. however, biocompatibility of nickel-titanium (ni-ti) alloy, which is the only practical sma at present, has been questioned because of its high nickel content. the aim of this study was to evaluate the ...	2015	25659946
aptamer decorated hyaluronan/chitosan nanoparticles for targeted delivery of 5-fluorouracil to muc1 overexpressing adenocarcinomas.	an aptamer (apt) conjugated hyaluronan/chitosan nanoparticles (hacsnps) were prepared as carrier for targeted delivery of 5-fluorouracil (5fu) to mucin1 (muc1) overexpressing colorectal adenocarcinomas. nanoparticles had about 181 nm size, encapsulation efficiency of 45.5 ± 2.8 and acceptable stability. conjugation of muc1-binding apt to the surface of the nanoparticles was confirmed by gel electrophoresis. toxicity and cellular uptake of nanoparticles were investigated by in vitro cytotoxicity ...	2015	25659689
voriconazole, an antifungal triazol that causes visual side effects, is an inhibitor of trpm1 and trpm3 channels.	administration of voriconazole, an antifungal triazole, causes transient visual disturbances in patients and attenuates the b-wave of the erg. we sought to identify the retinal target of voriconazole underlying the effect on the erg b-wave.	2015	25650413
receptor protein tyrosine phosphatase beta/zeta is a functional binding partner for vascular endothelial growth factor.	receptor protein tyrosine phosphatase beta/zeta (rptpβ/ζ) is a chondroitin sulphate (cs) transmembrane protein tyrosine phosphatase and is a receptor for pleiotrophin (ptn). rptpβ/ζ interacts with ανβ₃ on the cell surface and upon binding of ptn leads to c-src dephosphorylation at tyr530, β₃ tyr773 phosphorylation, cell surface nucleolin (ncl) localization and stimulation of cell migration. c-src-mediated β₃ tyr773 phosphorylation is also observed after vascular endothelial growth factor 165 (ve ...	2015	25644401
polymorphism of the xrcc3 gene and risk of gastric cancer in a kashmiri population: a case-control study.	dna repair plays a critical role in protecting the genome of the cell from the insults of cancer-causing agents. genetic polymorphisms in dna repair genes may influence individual variation in dna repair capacity, which may be associated with the risk of developing cancer. inherited polymorphisms of dna repair genes may contribute to variations in dna repair capacity and genetic susceptibility to different cancers. the x-ray repair complementing defective repair in chinese hamster cells 3 (xrcc3 ...	2015	25642794
a multi-pronged investigation into the effect of glucose starvation and culture duration on fed-batch cho cell culture.	in this study, omics-based analysis tools were used to explore the effect of glucose starvation and culture duration on monoclonal antibody (mab) production in fed-batch cho cell culture to gain better insight into how these parameters can be controlled to ensure optimal mab productivity and quality. titer and n-glycosylation of mabs, as well as proteomic signature and metabolic status of the production cells in the culture were assessed. we found that the impact of glucose starvation on the tit ...	2015	25899530
clonoamide, a new inhibitor of sterol o-acyltransferase, produced by clonostachys sp. bf-0131.	a new compound designated as clonoamide was isolated from a culture broth of the fungus clonostachys sp. bf-0131 by solvent extraction, diaion hp20 column chromatography, octadecylsilyl column chromatography and preparative hplc as an inhibitor of sterol o-acyltransferase (soat). the structure of clonoamide was elucidated as 2-oxo-9e,11e-tridecandienyl acetamide by various spectral analyses including nmr. the compound inhibited soat1 and soat2 isozymes with ic50 values of 39 and 110 μm, respecti ...	2015	25899123
antibody conjugates with unnatural amino acids.	antibody conjugates are important in many areas of medicine and biological research, and antibody-drug conjugates (adcs) are becoming an important next generation class of therapeutics for cancer treatment. early conjugation technologies relied upon random conjugation to multiple amino acid side chains, resulting in heterogeneous mixtures of labeled antibody. recent studies, however, strongly support the notion that site-specific conjugation produces a homogeneous population of antibody conjugat ...	2015	25898256
the criticality of high-resolution n-linked carbohydrate assays and detailed characterization of antibody effector function in the context of biosimilar development.	accurate measurement and functional characterization of antibody fc domain n-linked glycans is critical to successful biosimilar development. here, we describe the application of methods to accurately quantify and characterize the n-linked glycans of 2 igg1 biosimilars with effector function activity, and show the potential pitfalls of using assays with insufficient resolution. accurate glycan assessment was combined with glycan enrichment using lectin chromatography or production with glycosyla ...	2015	25898160
the antioxidant and antigenotoxic properties of citrus phenolics limonene and naringin.	phenolic compounds not only contribute to the sensory qualities of fruits and vegetables but also exhibit several health protective properties. limonene and naringin are the most popular phenolics found in citrus plants. in this study, we investigated the antioxidant capacities of limonene and naringin by the trolox equivalent antioxidant capacity (teac) assay and the cytotoxic effects by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (mtt) assay in chinese hamster fibroblast (v79) ...	2015	25896273
more similar than different: host cell protein production using three null cho cell lines.	to understand the diversity in the cell culture harvest (i.e., feedstock) provided for downstream processing, we compared host cell protein (hcp) profiles using three chinese hamster ovary (cho) cell lines in null runs which did not generate any recombinant product. despite differences in cho lineage, upstream process, and culture performance, the cell lines yielded similar cell-specific productivities for immunogenic hcps. to compare the dynamics of hcp production, we searched for correlations ...	2015	25894672
antidepressants activate the lysophosphatidic acid receptor lpa(1) to induce insulin-like growth factor-i receptor transactivation, stimulation of erk1/2 signaling and cell proliferation in cho-k1 fibroblasts.	different lines of evidence indicate that the lysophosphatidic acid (lpa) receptor lpa1 is involved in neurogenesis, synaptic plasticity and anxiety-related behavior, but little is known on whether this receptor can be targeted by neuropsychopharmacological agents. the present study investigated the effects of different antidepressants on lpa1 signaling. we found that in chinese hamster ovary (cho)-k1 fibroblasts expressing endogenous lpa1 tricyclic and tetracyclic antidepressants and fluoxetine ...	2015	25888927
sequencing the cho dxb11 genome reveals regional variations in genomic stability and haploidy.	the dhfr negative cho dxb11 cell line (also known as dux-b11 and dukx) was historically the first cho cell line to be used for large scale production of heterologous proteins and is still used for production of a number of complex proteins.	2015	25887056
radio-modifying potential of saraca indica against ionizing radiation: an in vitro study using chinese hamster lung fibroblast (v79) cells.	this study demonstrated the radioprotective efficacy of extracts prepared from stem bark of saraca indica (si) against x-rays induced cellular damage, which was evaluated by a battery of cytotoxicity, genotoxicity, apoptotic, and biochemical assays using chinese hamster fibroblast (v79) cells. cell viability and surviving fraction were increased significantly when v79 cells were preconditioned with optimal concentration of hydroalcoholic extract (he; 50 μg/ml) of si for 2 h prior exposure to x-r ...	2015	25879585
'bioquart' project: design of a novel in situ protocol for the simultaneous visualisation of chromosomal aberrations and micronuclei after irradiation at microbeam facilities.	the aim of the 'bioquart' (biologically weighted quantities in radiotherapy) project is to develop measurement techniques for characterising charged particle track structure on different length scales, and to correlate at the cellular level the track structure properties with the biological effects of radiation. this multi-scale approach will allow characterisation of the radiation qualities used in radiotherapy and the related biological effects. charged-particle microbeam facilities were chose ...	2015	25877532
oxidized ldl (oxldl) activates the angiotensin ii type 1 receptor by binding to the lectin-like oxldl receptor.	the angiotensin ii type 1 receptor (at1) is a 7-transmembrane domain gpcr that when activated by its ligand angiotensin ii, generates signaling events promoting vascular dysfunction and the development of cardiovascular disease. here, we show that the single-transmembrane oxidized ldl (oxldl) receptor (lox-1) resides in proximity to at1 on cell-surface membranes and that binding of oxldl to lox-1 can allosterically activate at1-dependent signaling events. oxldl-induced signaling events in human ...	2015	25877213
production of α2,6-sialylated igg1 in cho cells.	the presence of α2,6-sialic acids on the fc n-glycan provides anti-inflammatory properties to the iggs through a mechanism that remains unclear. fc-sialylated iggs are rare in humans as well as in industrial host cell lines such as chinese hamster ovary (cho) cells. facilitated access to well-characterized α2,6-sialylated iggs would help elucidate the mechanism of this intriguing igg's effector function. this study presents a method for the efficient fc glycan α2,6-sialylation of a wild-type and ...	2015	25875452
a tetravalent bispecific tandab (cd19/cd3), afm11, efficiently recruits t cells for the potent lysis of cd19(+) tumor cells.	to harness the potent tumor-killing capacity of t cells for the treatment of cd19(+) malignancies, we constructed afm11, a humanized tetravalent bispecific cd19/cd3 tandem diabody (tandab) consisting solely of fv domains. the molecule exhibits good manufacturability and stability properties. afm11 has 2 binding sites for cd3 and 2 for cd19, an antigen that is expressed from early b cell development through differentiation into plasma cells, and is an attractive alternative to cd20 as a target fo ...	2015	25875246
antibody glycosylation and its impact on the pharmacokinetics and pharmacodynamics of monoclonal antibodies and fc-fusion proteins.	understanding the impact of glycosylation and keeping a close control on glycosylation of product candidates are required for both novel and biosimilar monoclonal antibodies (mabs) and fc-fusion protein development to ensure proper safety and efficacy profiles. most therapeutic mabs are of igg class and contain a glycosylation site in the fc region at amino acid position 297 and, in some cases, in the fab region. for fc-fusion proteins, glycosylation also frequently occurs in the fusion partners ...	2015	25872915
fluorescence dye-based detection of mab aggregates in cho culture supernatants.	product yields, efficacy, and safety of monoclonal antibodies (mabs) are reduced by the formation of higher molecular weight aggregates during upstream processing. in-process characterization of mab aggregate formation is a challenge since there is a lack of a fast detection method to identify mab aggregates in cell culture. in this work, we present a rapid method to characterize mab aggregate-containing chinese hamster ovary (cho) cell culture supernatants. the fluorescence dyes thioflavin t (t ...	2015	25869484
one-step generation of triple knockout cho cell lines using crispr/cas9 and fluorescent enrichment.	the crispr/cas9 genome editing technology has previously been shown to be a highly efficient tool for generating gene disruptions in cho cells. in this study we further demonstrate the applicability and efficiency of crispr/cas9 genome editing by disrupting fut8, bak and bax simultaneously in a multiplexing setup in cho cells. to isolate cas9-expressing cells from transfected cell pools, gfp was linked to the cas9 nuclease via a 2a peptide. with this method, the average indel frequencies generat ...	2015	25864574
a quantitative proteomic analysis of cellular responses to high glucose media in chinese hamster ovary cells.	a goal in recombinant protein production using chinese hamster ovary (cho) cells is to achieve both high specific productivity and high cell density. addition of glucose to the culture media is necessary to maintain both cell growth and viability. we varied the glucose concentration in the media from 5 to 16 g/l and found that although specific productivity of cho-dg44 cells increased with the glucose level, the integrated viable cell density decreased. to examine the biological basis of these r ...	2015	25857574
is nitrous oxide a genotoxic carcinogen?	nitrous oxide (n2o) has been widely used as a dental and surgical anaesthetic for over 150 years. however, results from a recent study suggested that increased dna damage was seen in lymphocytes from surgical patients and this led to its continued clinical use to be questioned. the data can be challenged on technical grounds and must be considered with other studies in order to assess any possible risk. there are other studies indicating that n2o has weak genotoxicity in man, but these are confu ...	2015	25852088
a direct qpcr method for residual dna quantification in monoclonal antibody drugs produced in cho cells.	chinese hamster ovary (cho) cells are the host cell of choice for manufacturing of monoclonal antibody (mab) drugs in the biopharmaceutical industry. host cell dna is an impurity of such manufacturing process and must be controlled and monitored in order to ensure drug purity and safety. a conventional method for quantification of host residual dna in drug requires extraction of dna from the mab drug substance with subsequent quantification of the extracted dna using real-time pcr (qpcr). here w ...	2015	25850374
mechanism of action studies of lomaiviticin a and the monomeric lomaiviticin aglycon. selective and potent activity toward dna double-strand break repair-deficient cell lines.	(-)-lomaiviticin a (1) and the monomeric lomaiviticin aglycon [aka: (-)-mk7-206, (3)] are cytotoxic agents that induce double-strand breaks (dsbs) in dna. here we elucidate the cellular responses to these agents and identify synthetic lethal interactions with specific dna repair factors. toward this end, we first characterized the kinetics of dna damage by 1 and 3 in human chronic myelogenous leukemia (k562) cells. dsbs are rapidly induced by 3, reaching a maximum at 15 min post addition and are ...	2015	25849366
benchmarking of commercially available cho cell culture media for antibody production.	in this study, eight commercially available, chemically defined chinese hamster ovary (cho) cell culture media from different vendors were evaluated in batch culture using an igg-producing cho dg44 cell line as a model. medium adaptation revealed that the occurrence of even small aggregates might be a good indicator of cell growth performance in subsequent high cell density cultures. batch experiments confirmed that the culture medium has a significant impact on bioprocess performance, but high ...	2015	25846330
synthesis and pharmacology of halogenated δ-opioid-selective [d-ala(2)]deltorphin ii peptide analogues.	deltorphins are naturally occurring peptides produced by the skin of the giant monkey frog (phyllomedusa bicolor). they are δ-opioid receptor-selective agonists. herein, we report the design and synthesis of a peptide, tyr-d-ala-(pi)phe-glu-ile-ile-gly-nh2 3 (gate3-8), based on the [d-ala(2)]deltorphin ii template, which is δ-selective in in vitro radioligand binding assays over the μ- and κ-opioid receptors. it is a full agonist in [(35)s]gtpγs functional assays and analgesic when administered ...	2015	25844930
negative cooperativity across β1-adrenoceptor homodimers provides insights into the nature of the secondary low-affinity cgp 12177 β1-adrenoceptor binding conformation.	at the β1-adrenoceptor, cgp 12177 potently antagonizes agonist responses at the primary high-affinity catecholamine conformation while also exerting agonist effects of its own through a secondary low-affinity conformation. a recent mutagenesis study identified transmembrane region (tm)4 of the β1-adrenoceptor as key for this low-affinity conformation. others suggested that tm4 has a role in β1-adrenoceptor oligomerization. here, assessment of the dissociation rate of a fluorescent analog of cgp ...	2015	25837585
pcb 77 action in ovary cells--toxic effects, apoptosis induction and cell cycle analysis.	pcb 77 (3,3',4,4'-tetrachlorobiphenyl), a non-ortho congener with planar configuration, has been identified as potential endocrine disrupter capable to increase the risk of reproductive and developmental failure.	2015	25835094
evaluating the interaction between ucoe and dhfr-linked amplification and stability of recombinant protein expression.	chinese hamster ovary (cho) cells are widely used in the biopharmaceutical industry. in the creation of mammalian cell lines plasmid dna carrying the gene-of-interest integrates randomly into the host cell genome, which results in variable levels of gene expression between cell lines due to gene silencing mechanisms. in addition, cell lines often show unstable protein production during long-term culture. this means that a large number of clones need to be screened in order to isolate stable, hig ...	2015	25829363
unveiling the principle of microrna-mediated redundancy in cellular pathway regulation.	understanding the multifaceted nature of microrna (mirna) function in mammalian cells is still a challenge. commonly accepted principles of cooperativity and multiplicity of mirna function imply that individual mrnas can be targeted by several mirnas whereas a single mirna may concomitantly regulate a subset of different genes. however, there is a paucity of information whether multiple mirnas regulate critical cellular events and thereby acting redundantly. to gain insight into this notion, we ...	2015	25826657
generic raman-based calibration models enabling real-time monitoring of cell culture bioreactors.	raman-based multivariate calibration models have been developed for real-time in situ monitoring of multiple process parameters within cell culture bioreactors. developed models are generic, in the sense that they are applicable to various products, media, and cell lines based on chinese hamster ovarian (cho) host cells, and are scalable to large pilot and manufacturing scales. several batches using different cho-based cell lines and corresponding proprietary media and process conditions have be ...	2015	25825868
2-d western blotting for evaluation of antibodies developed for detection of host cell protein.	recombinant proteins generated for therapeutic use must be substantially free of residual host cell protein (hcp). the presence of host cell protein (hcp) is usually assayed by elisa using a polyclonal antibody mixture raised against a population of proteins derived from the host cell background. this antibody should recognize as high a proportion as possible of the potential hcps in a given sample. a recommended method for evaluating the assay involves two-dimensional electrophoretic separation ...	2015	25820736
carbamates: a study on genotoxic, cytotoxic, and apoptotic effects induced in chinese hamster ovary (cho-k1) cells.	in vitro effects of the carbamates pirimicarb and zineb and their formulations aficida® (50% pirimicarb) and azzurro® (70% zineb), respectively, were evaluated in chinese hamster ovary (cho-k1) cells. whereas the cytokinesis-blocked micronucleus cytome assay was employed to test for genotoxicity, mtt, neutral red (nr), and apoptosis evaluation were used as tests for estimating cell viability and succinic dehydrogenase activity, respectively. concentrations tested were 10-300 μg/ml for pirimicarb ...	2015	25820133
cerebral visual impairment and intellectual disability caused by pgap1 variants.	homozygous variants in pgap1 (post-gpi attachment to proteins 1) have recently been identified in two families with developmental delay, seizures and/or spasticity. pgap1 is a member of the glycosylphosphatidylinositol anchor biosynthesis and remodeling pathway and defects in this pathway are a subclass of congenital disorders of glycosylation. here we performed whole-exome sequencing in an individual with cerebral visual impairment (cvi), intellectual disability (id), and factor xii deficiency ...	2015	25804403
effects of disruption of the nucleotide pattern in crid element and kozak sequence of interferon β on mrna stability and protein production.	interferon β (ifnβ) is the most important drug that has been used frequently for multiple sclerosis treatment. this study has tried to improve the ifnβ production by introducing mutations in the coding region of ifnβ, while its amino acid sequence is intact. two recombinant vectors ifnβ(k) and ifnβ(k+crid )were designed by site-directed mutagenesis. the ifnβ(k) and ifnβ(k+crid) have two substitutions in kozak sequence and four substitutions in crid sequence, respectively. the chinese hamster ova ...	2015	25799196
rhenium(i) polypyridine dibenzocyclooctyne complexes as phosphorescent bioorthogonal probes: synthesis, characterization, emissive behavior, and biolabeling properties.	we report the development of rhenium(i) polypyridine complexes appended with a dibenzocyclooctyne (dibo) moiety as bioorthogonal probes for azide-modified biomolecules. three phosphorescent rhenium(i) polypyridine dibo complexes [re(n^n)(co)3(py-c6-dibo)][cf3so3] (py-c6-dibo=3-(n-(6-(3,4:7,8-dibenzocyclooctyne-5-oxycarbonylamino)hexyl)aminocarbonyl)pyridine; n^n=1,10-phenanthroline (phen) (1a), 3,4,7,8-tetramethyl-1,10-phenanthroline (me4-phen) (2a), 4,7-diphenyl-1,10-phenanthroline (ph2-phen) ( ...	2015	25798989
metabolic markers associated with high mannose glycan levels of therapeutic recombinant monoclonal antibodies.	high mannose (hm) glycan levels on secreted monoclonal antibodies can be influenced by external factors, including osmolality and copper deficiency, and by intrinsic factors determined by different cell lines. in order to identify the metabolic markers associated with hm glycan levels, metabolomics analysis was performed to assess the changes in the extracellular metabolites of recombinant cell lines at different time points during fed-batch production process. ornithine was identified as the co ...	2015	25797592
in vitro comparison of duration of action of melatonin agonists on melatonin mt1 receptor: possible link between duration of action and dissociation rate from receptor.	melatonin mt1 and mt2 receptors are gi protein-coupled receptors and promising therapeutic targets for a number of diseases. a proportion of g protein-coupled receptor agonists and antagonists have been classified according to their duration of action, which influences their pharmacological efficacy. however, the duration of action of melatonin agonists remains unclear. in this study, we investigated the duration of action of melatonin agonists (melatonin, 2-iodomelatonin, ramelteon, and the ram ...	2015	25797281
the recombinant equine lhβ subunit combines divergent intracellular traits of human lhβ and cgβ subunits.	the pituitary lhβ and placental cgβ subunits are products of different genes in primates. the major structural difference between the two subunits is in the carboxy-terminal region, where the short carboxyl sequence of hlhβ is replaced by a longer o-glycosylated carboxy-terminal peptide in hcgβ. in association with this structural deviation, there are marked differences in the secretion kinetics and polarized routing of the two subunits. in equids, however, the cgβ and lhβ subunits are products ...	2015	25796287
reduced expression of the oligosaccharyltransferase exacerbates protein hypoglycosylation in cells lacking the fully assembled oligosaccharide donor.	a defect in the assembly of the oligosaccharide donor (dol-pp-glcnac(2)man(9)glc(3)) for n-linked glycosylation causes hypoglycosylation of proteins by the oligosaccharyltransferase (ost). mammalian cells express two ost complexes that have different catalytic subunits (stt3a or stt3b). we monitored glycosylation of proteins in asparagine-linked glycosylation 6 (alg6) deficient cell lines that assemble dol-pp-glcnac(2)man(9) as the largest oligosaccharide donor. based upon pulse labeling experim ...	2015	25792706
optimization of cell line development in the gs-cho expression system using a high-throughput, single cell-based clone selection system.	therapeutic antibodies are commonly produced by high-expressing, clonal and recombinant chinese hamster ovary (cho) cell lines. currently, cho cells dominate as a commercial production host because of their ease of use, established regulatory track record, and safety profile. cho-k1sv is a suspension, protein-free-adapted cho-k1-derived cell line employing the glutamine synthetase (gs) gene expression system (gs-cho expression system). the selection of high-producing mammalian cell lines is a cr ...	2015	25792187
effects of targeted phosphorylation site mutations in the dna-pkcs phosphorylation domain on low and high let radiation sensitivity.	the present study investigated the effect of targeted mutations in the dna-dependent protein kinase catalytic subunit and phosphorylation domains on the survival of cells in response to different qualities of ionizing radiation. mutated chinese hamster ovary v3 cells were exposed to 500 mev/nucleon initial energy and 200 kev/μm monoenergetic fe ions; 290 mev/nucleon initial energy and average 50 kev/μm spread-out bragg peak c ions; 70 mev/nucleon initial energy and 1 kev/μm monoenergetic protons ...	2015	25789011
enhanced anticancer activity of 5-fu in combination with bestatin: evidence in human tumor-derived cell lines and an h22 tumor-bearing mouse.	the clinical use of 5-fluorouracil (5-fu) is increasingly limited by low response rates, adverse reactions, and toxicity. a drug combination offers a new strategy for appropriate use of 5-fu. bestatin, an aminopeptidase n (apn) inhibitor, has been used as an adjuvant chemotherapy drug because of its actions to suppress tumorigenesis and invasion. the current study evaluated the anticancer efficacy of 5-fu plus bestatin at the cellular and animal level. the combination killed more colonic cancer, ...	2015	25788051
characterization of russell bodies accumulating mutant antithrombin derived from the endoplasmic reticulum.	the endoplasmic reticulum (er) adjusts its size and architecture to adapt to change in the surrounding environment. russell bodies (rbs) were originally described as dilated structures of the er cisternae containing large amounts of mutant immunoglobulin. similar structures are observed in a wide variety of mutant proteins accumulated in the er. we previously prepared chinese hamster ovary (cho) cells in which the expression of mutant antithrombin (at) (c95r) was controlled with a tet-on system ...	2015	25787801
increased production of soluble vascular endothelial growth factors receptor-1 in cho-cell line by using new combination of chitosan-protein lipid nanoparticles.	the soluble vascular endothelial growth factor receptor-1 (vegfr1) or sflt-1 has important role in antiangiogenesis. in this study, the increase expression and production of sflt-1 fragment by newly designed chpl-nps nanoparticles (chitosan-protein lipid) using chinese hamster ovary cell line (cho) was evaluated. the assessment and purification of sflt-1 were carried out by western blotting and fast protein liquid chromatography (fplc). thereafter, the angiostatic effect of gene transfer of sflt ...	2015	25785168
pkc-mediated inhibitory feedback of the cholecystokinin 1 receptor controls the shape of oscillatory ca²⁺ signals.	translation of extracellular hormonal input into cellular responses is often mediated by repetitive increases in cytosolic free ca(2+) concentration ([ca(2+) ]c ). amplitude, duration and frequency of these so-called [ca(2+) ]c oscillations then carry information about the nature and concentration of the extracellular signalling molecule. at present, there are different hypotheses concerning the induction and control of these oscillations. here, we investigated the role of agonist-induced recept ...	2015	25779353
dose-exposure proportionality of a novel recombinant follicle-stimulating hormone (rfsh), fe 999049, derived from a human cell line, with comparison between caucasian and japanese women after subcutaneous administration.	fe 999049 is a novel recombinant follicle-stimulating hormone (rfsh) preparation expressed by a human cell line (per.c6(®)), in contrast to existing rfsh preparations expressed by chinese hamster ovary (cho) cell lines. since the individual dose of rfsh may be altered depending on the response in women undergoing assisted reproductive technologies, knowledge on the dose-exposure linearity and proportionality is important. the purpose of these studies was to investigate the dose-exposure linearit ...	2015	25773354
bryostatin-1 causes radiosensitization of bmg-1 malignant glioma cells through differential activation of protein kinase-cδ not evident in the non-malignant aa8 fibroblasts.	bryostatin-1 (bryo-1), a non-phorbol ester, is known to sensitize mammalian cells against certain chemotherapeutic drugs. we assessed its ability to modify radiation response of mammalian cells using chinese hamster fibroblasts aa8 cells and human malignant glioma bmg-1 cells. in the malignant glioma bmg-1 cell line, bryo-1 pre-treatment significantly enhanced radiation-induced growth inhibition and cytogenetic damage, and further reduced the clonogenic cell survival as compared to cells irradia ...	2015	25472878
effect of temperature shift on levels of acidic charge variants in igg monoclonal antibodies in chinese hamster ovary cell culture.	during the production of therapeutic monoclonal antibodies (mabs), not only enhancement of mab productivity but also control of quality attributes is critical. charge variants, which are among the most important quality attributes, can substantially affect the in vitro and in vivo properties of mabs. during process development for the production of mabs in a chinese hamster ovary cell line, we have observed that an improvement in mab titer is accompanied by an increase in the content of acidic c ...	2015	25466646
biodirected synthesis of miconazole-conjugated bacterial silver nanoparticles and their application as antifungal agents and drug delivery vehicles.	the recent strategy to improve the efficacy of drugs is to combine them with metal nanoparticles for the control of microbial infections. considering this fact, we developed a low cost and eco-friendly method for silver nanoparticles synthesis using the cell free supernatant of delftia sp. strain kcm-006 and their application as antifungal agents and as a drug carrier. transmission electron microscopy (tem) and dynamic light scattering (dls) analysis revealed the formation of spherical and monod ...	2015	25460601
accumulation of carboxy-terminal fragments of app increases phosphodiesterase 8b.	the long-standing "amyloid hypothesis" that alzheimer's disease is caused by the production and aggregation of amyloid-β faces serious challenges by data recently obtained from neuroimaging studies and amyloid-β amyloid-focused clinical trials. meanwhile, accumulation of carboxy-terminal fragments (ctfs) of the amyloid precursor protein (app) may be neurotoxic and may impair synaptic plasticity and long-term memory in alzheimer's disease, as suggested in murine models. to clarify these issues, w ...	2015	25457556
hep-2 cell based indirect immunofluorescence assay for antinuclear antibodies as a potential diagnosis of drug-induced autoimmunity in nonclinical toxicity testing.	antinuclear antibodies (anas) are important biomarkers in the diagnosis of autoimmune diseases in humans; however, the diagnostic performance of ana in nonclinical safety studies are not well understood. here, we studied the use of anas as potential nonclinical biomarkers for drug-induced autoimmunity (dia) using a hep-2 based indirect immunofluorescence assay (ifa). initially, mrl-fas(lpr)/j mice and hgcl₂-treated rats were used as sle-positive models. serum samples obtained from 94 normal mice ...	2015	25455225
titer of trastuzumab produced by a chinese hamster ovary cell line is associated with tricarboxylic acid cycle activity rather than lactate metabolism.	achieving high productivity and quality is the final goal of therapeutic antibody development, but the productivity and quality of antibodies are known to be substantially dependent on the nature of the cell lines expressing the antibodies. we characterized two contrasting cell lines that produce trastuzumab, namely cell line a with a high titer and a low aggregate content and cell line b with a low titer and a high aggregate content to identify the causes of the differences. we observed the fol ...	2015	25449760
conserved microrna function as a basis for chinese hamster ovary cell engineering.	the use of micrornas (mirnas) for improving the efficiency of recombinant protein production by cho cells is gaining considerable interest for their ability to regulate entire molecular networks. differential mirna expression profiling and large-scale transient screening have been the prerequisite for the selection of mirna candidates for stable manipulation, reported in cho cells expressing a range of recombinant products. we selected a potent and well characterised tumour suppressor mirna, mir ...	2015	25527440
targeting ß2 adrenergic receptors regulate human t cell function directly and indirectly.	it is well-established that central nervous system activation affects peripheral blood mononuclear cell (pbmcs) function through the release of the catecholamines (epi) and norepinephrine (ne), which act on ß2-adrenergic receptors (ß2ar). however, most studies have used non-specific stimulation of cells rather than antigen-specific responses. likewise, few studies have parsed out the direct effects of ß2ar stimulation on t cells versus indirect effects via adrenergic stimulation of antigen prese ...	2015	25526818
fty720 phosphate activates sphingosine-1-phosphate receptor 2 and selectively couples to gα12/13/rho/rock to induce myofibroblast contraction.	fty720 phosphate (fty720-p; 2-amino-2-[2-(4-octylphenyl)ethyl]-1,3-propanediol, monodihydrogen phosphate ester) is a nonselective sphingosine-1-phosphate (s1p) receptor agonist thought to be devoid of activity at the s1p2 receptor subtype. however, we have recently shown that fty720-p displays significant s1p2 receptor agonist activity in recombinant cells and fibroblasts expressing endogenous s1p2 receptors. to elucidate the s1p2-dependent signaling pathways that were activated by fty720-p, we ...	2015	25762025
thromboxane a2 receptor polymorphism in association with cerebral infarction and its regulation on platelet function.	binding of thromboxane a2 (txa2) to its receptor txa2r modulates thrombosis/hemostasis and plays a vital role in the pathogenesis of cerebral infarction (ci). in this study, we investigated the relationship between txa2r polymorphisms and ci in chinese han population and the effect on platelet function by these polymorphisms. polymerase chain reaction and ligase detection reaction (pcr-ldr) was performed in 230 ci patients and 143 healthy volunteers to examine four single nucleotide polymorphism ...	2015	25557379
evaluation of gambierol and its analogs for their inhibition of human kv1.2 and cytotoxicity.	gambierol and its heptacyclic and tetracyclic analogs were tested for inhibitory activity against the human voltage-gated potassium channel kv1.2 (hkv1.2), which was stably expressed in chinese hamster ovary (cho) cells. gambierol, the heptacyclic analog, and the tetracyclic analog inhibited the potassium current evoked by a step pulse from -80mv to 40mv. the ic50 values for the three compounds were 0.75±0.15nm, 7.6±1.2nm, and 28±4.0nm (the mean±sem, n=3), respectively. the cytotoxic activity wa ...	2015	25556093
carboxylated short single-walled carbon nanotubes but not plain and multi-walled short carbon nanotubes show in vitro genotoxicity.	long carbon nanotubes (cnts) resemble asbestos fibers due to their high length to diameter ratio and they thus have genotoxic effects. another parameter that might explain their genotoxic effects is contamination with heavy metal ions. on the other hand, short (1-2 µm) cnts do not resemble asbestos fibers, and, once purified from contaminations, they might be suitable for medical applications. to identify the role of fiber thickness and surface properties on genotoxicity, well-characterized shor ...	2015	25505129
mid-infrared spectroscopy-based analysis of mammalian cell culture parameters.	within the framework of process analytical technology, infrared spectroscopy (ir) has been used for characterization of biopharmaceutical production processes. although noninvasive attenuated total reflection (atr) spectroscopy can be regarded as gold standard within ir-based process analytics, simpler and more cost-effective mid-infrared (mir) instruments might improve acceptability of this technique for high-level monitoring of small scale experiments as well as for academia where financial re ...	2015	25504543
expression of difficult-to-remove host cell protein impurities during extended chinese hamster ovary cell culture and their impact on continuous bioprocessing.	during biopharmaceutical manufacturing, chinese hamster ovary (cho) cells produce hundreds of extracellular host cell protein (hcp) impurities, which must be removed from the therapeutic product by downstream purification operations to ensure patient safety. a subset of 118 of these hcps have been reported as exceptionally difficult to remove during downstream purification because they co-purify due to retention characteristics on chromatographic media and/or product-association through strongly ...	2015	25502542
direct site-specific glycoform identification and quantitative comparison of glycoprotein therapeutics: imiglucerase and velaglucerase alfa.	gaucher disease, the most common lysosomal metabolic disorder, can be treated with enzyme replacement therapy (ert). recombinant human glucocerebrosidase imiglucerase (cerezyme(®)), produced in chinese hamster ovary cells, has been used for ert of gaucher disease for 20 years. another recombinant glucocerebrosidase velaglucerase alfa (vpriv), expressed in a human fibroblast cell line, was approved by the us food and drug administration in 2010. the amino acid sequence difference at residue 495 o ...	2015	25501675
plasmid transfection in mammalian cells spatiotemporally tracked by a gold nanoparticle.	recent advances in cell transfection have suggested that delivery of a gene on a gold nanoparticle (aunp) can enhance transfection efficiency. the mechanism of transfection is poorly understood, particularly when the gene is appended to a aunp, as expression of the desired exogenous protein is dependent not only on the efficiency of the gene being taken into the cell but also on efficient endosomal escape and cellular processing of the nucleic acid. design of a multicolor surface energy transfer ...	2015	25494916
evaluation of the cytotoxic and mutagenic potential of three ginkgolic acids.	ginkgolic acids (gas) are alkylphenols which can be found in the fruits and leaves of ginkgo biloba l. (ginkgoaceae) used in herbal teas, drugs and food supplements. standardized leaf extracts of g. biloba are widely used in the therapy of cognitive decline including alzheimer's diseases. however, gas are known to have cytotoxic and allergenic potential and are suspected to possess genotoxic properties. therefore, we examined in this study the cytotoxicity and mutagenicity of three major gas wit ...	2015	25448085
low dosage of rimonabant leads to anxiolytic-like behavior via inhibiting expression levels and g-protein activity of kappa opioid receptors in a cannabinoid receptor independent manner.	there is an increasing number of studies demonstrating the direct effect of the cannabinoid receptor 1 (cb1) antagonist/inverse agonist rimonabant on the opioid system. the kappa opioid receptors (kors) are well known to mediate depression- and anxiety-like behavior. clinical studies on chronic rimonabant administration have revealed that rimonabant leads to a very similar pathophysiology, suggesting a potential impact of rimonabant on kors.	2015	25446673