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accelerated homology-directed targeted integration of transgenes in chinese hamster ovary cells via crispr/cas9 and fluorescent enrichment.targeted gene integration into site-specific loci can be achieved in chinese hamster ovary (cho) cells via crispr/cas9 genome editing technology and the homology-directed repair (hdr) pathway. the low efficiency of hdr often requires antibiotic selection, which limits targeted integration of multiple genes at multiple sites. to improve hdr-mediated targeted integration, while avoiding the use of selection markers, chemical treatment for increased hdr, and fluorescent enrichment of genome-edited ...201627159230
apolipoprotein e lipoprotein particles inhibit amyloid-β uptake through cell surface heparan sulphate proteoglycan.the accumulation, aggregation and deposition of amyloid-β (aβ) peptides in the brain are central to the pathogenesis of alzheimer's disease (ad). alzheimer's disease risk increases significantly in individuals carrying one or two copies of apoe ε4 allele compared to individuals with an ε3/ε3 genotype. growing evidence has demonstrated that apolipoprotein e (apoe) strongly influences ad pathogenesis by controlling aβ aggregation and metabolism. heparan sulphate proteoglycans (hspgs) are abundant ...201627151330
highly luminescent heterostructured copper-doped zinc sulfide nanocrystals for application in cancer cell labeling.the structural characteristics of the seed-mediated synthesis of heterostructured cus-zns nanocrystals (ncs) and cu-doped zns (zns:cu) ncs synthesized by two different protocols are compared and analyzed. at high cu dopant concentrations, segregated subclusters of zns and cus are observed. the photoluminescence quantum yield of zns:cu ncs is about 50-80 %; a value much higher than that of zns ncs (6 %). finally, these ncs are coated with a thin silica shell by using (3-mercaptopropyl)triethoxysi ...201627146419
classification of normal and apoptotic cells from fluorescence microscopy images using generalized polynomial chaos and level set function.accurate automated quantitative analysis of living cells based on fluorescence microscopy images can be very useful for fast evaluation of experimental outcomes and cell culture protocols. in this work, an algorithm is developed for fast differentiation of normal and apoptotic viable chinese hamster ovary (cho) cells. for effective segmentation of cell images, a stochastic segmentation algorithm is developed by combining a generalized polynomial chaos expansion with a level set function-based se ...201627142234
expression and bioactivity of recombinant human serum albumin and dtmp fusion proteins in cho cells.the 14-amino acid (iegptlrqwlaara) thrombopoietin mimetic peptide (tmp) shares no sequence homology with native thrombopoietin (tpo). when dimerized, it displays a high-binding affinity for the tpo receptor and has equipotent bioactivity with recombinant human tpo (rhtpo) in stimulating proliferation and maturation of megakaryocytes in vitro. however, tmp is limited for clinical usage because of its short half-life in vivo. in this study, fusion proteins that composed of tandem dimer of tmp (dtm ...201627115755
disruption of the gene c12orf35 leads to increased productivities in recombinant cho cell lines.recently, we reported that the loss of a telomeric region of chromosome 8 in chinese hamster ovary (cho) cells correlates with higher recombinant productivities. new cell lines lacking this region, called cho-c8del, showed several advantages during cell line generation and for the production of recombinant proteins (ritter et al., 2016, biotechnol bioeng). here, we performed knock-down and knock-out experiments of genes located within this telomeric region of chromosome 8 to identify the genes c ...201627183150
expression and characterization of a potent long-acting glp-1 receptor agonist, glp-1-igg2σ-fc.human glp-1 (glucagon-like peptide-1) can produce a remarkable improvement in glycemic control in patients with type 2 diabetes. however, its clinical benefits are limited by its short half-life, which is less than 2 min because of its small size and rapid enzymatic inactivation by dipeptidyl peptidase iv. we engineered glp-1-igg2σ-fc, a 68-kda fusion protein linking a variant human glp-1 (a8g/g26e/r36g) to a human igg2σ constant heavy-chain. a stably transfected chinese hamster ovary cell line ...201627232339
moderate and strong static magnetic fields directly affect egfr kinase domain orientation to inhibit cancer cell proliferation.static magnetic fields (smfs) can affect cell proliferation in a cell-type and intensity-dependent way but the mechanism remains unclear. at the same time, although the diamagnetic anisotropy of proteins has been proposed decades ago, the behavior of isolated proteins in magnetic fields has not been directly observed. here we show that smfs can affect isolated proteins at the single molecular level in an intensity-dependent manner. we found that epidermal growth factor receptor (egfr), a protein ...201627223425
mutation detection in an antibody-producing chinese hamster ovary cell line by targeted rna sequencing.chinese hamster ovary (cho) cells have been used widely in the pharmaceutical industry for production of biological therapeutics including monoclonal antibodies (mab). the integrity of the gene of interest and the accuracy of the relay of genetic information impact product quality and patient safety. here we employed next-generation sequencing, particularly rna-seq, and developed a method to systematically analyze the mutation rate of the mrna of cho cell lines producing a mab. the effect of an ...201627088091
dimerization of melanocortin receptor 1 (mc1r) and mc5r creates a ligand-dependent signal modulation: potential participation in physiological color change in the flounder.vertebrates produce α-melanocyte-stimulating hormone (α-msh), which contains an n-terminal acetyl group, and desacetyl-α-msh, which does not contain an n-terminal acetyl group. in teleosts and amphibians, α-msh-related peptides stimulate pigment dispersion via melanocortin receptors 1-5 (mc1r-mc5r), which are members of the g-protein-coupled receptor (gpcr) family. we previously reported an interesting phenomenon associated with physiological color changes in the skin of a flatfish, barfin floun ...201627080548
experimental evaluation of proposed small-molecule inhibitors of water channel aquaporin-1.the aquaporin-1 (aqp1) water channel is a potentially important drug target, as aqp1 inhibition is predicted to have therapeutic action in edema, tumor growth, glaucoma, and other conditions. here, we measured the aqp1 inhibition efficacy of 12 putative small-molecule aqp1 inhibitors reported in six recent studies, and one aqp1 activator. osmotic water permeability was measured by stopped-flow light scattering in human and rat erythrocytes that natively express aqp1, in hemoglobin-free membrane ...201626993802
identification of drug-like inhibitors of insulin-regulated aminopeptidase through small-molecule screening.intracerebroventricular injection of angiotensin iv, a ligand of insulin-regulated aminopeptidase (irap), has been shown to improve cognitive functions in several animal models. consequently, irap is considered a potential target for treatment of cognitive disorders. to identify nonpeptidic irap inhibitors, we adapted an established enzymatic assay based on membrane preparations from chinese hamster ovary cells and a synthetic peptide-like substrate for high-throughput screening purposes. the 38 ...201627078680
the long intron 1 of growth hormone gene from reeves' turtle (chinemys reevesii) correlates with negatively regulated gh expression in four cell lines.turtles grow slowly and have a long lifespan. ultrastructural studies of the pituitary gland in reeves' turtle (chinemys reevesii) have revealed that the species possesses a higher nucleoplasmic ratio and fewer secretory granules in growth hormone (gh) cells than other animal species in summer and winter. c. reevesii gh gene was cloned and species-specific similarities and differences were investigated. the full gh gene sequence in c. reevesii contains 8517 base pairs (bp), comprising five exons ...201627077853
one-step conjugation method for site-specific antibody-drug conjugates through reactive cysteine-engineered antibodies.engineered cysteine residues are particularly convenient for site-specific conjugation of antibody-drug conjugates (adc), because no cell engineering and additives are required. usually, unpaired cysteine residues form mixed disulfides during fermentation in chinese hamster ovarian (cho) cells; therefore, additional reduction and oxidization steps are required prior to conjugation. in this study, we prepared light chain (lc)-q124c variants in igg and examined the conjugation efficiency. intrigui ...201627074832
comprehensive genome and epigenome characterization of cho cells in response to evolutionary pressures and over time.the most striking characteristic of cho cells is their adaptability, which enables efficient production of proteins as well as growth under a variety of culture conditions, but also results in genomic and phenotypic instability. to investigate the relative contribution of genomic and epigenetic modifications towards phenotype evolution, comprehensive genome and epigenome data are presented for six related cho cell lines, both in response to perturbations (different culture conditions and media a ...201627072894
quantitative analysis of the supernatant from host and transfected cho cells using itraq 8-plex technique.we employed uplc-ms/ms with itraq 8-plex labeling to quantitatively analyze the supernatant produced by two chinese hamster ovary (cho) cell lines (cho k1sv and cho cat-s). in each case, the supernatant from the host and three transfected clones were analyzed at days 5, 7, and 10 of culture. a total of eight itraq 8-plex experiments were performed. for each cell line, the overlap of supernatant protein identifications between transfected clones is over 60%. over 70% of the supernatant proteins i ...201627070921
isotetrandrine reduces astrocyte cytotoxicity in neuromyelitis optica by blocking the binding of nmo-igg to aquaporin 4.neuromyelitis optica (nmo) is a severe neurological demyelinating autoimmune disease that affects the optic nerves and spinal cord with no cure and no fda-approved therapy. research over the last decade revealed that the binding of nmo-igg to the water channel protein astrocyte aquaporin 4 (aqp4) might be the primary cause of nmo pathogenesis. the purpose of this study was to identify potential blockers of nmo-igg and aqp4 binding.201627064690
characterization of recombinant human diamine oxidase (rhdao) produced in chinese hamster ovary (cho) cells.human diamine oxidase (hdao) efficiently degrades polyamines and histamine. reduced enzyme activities might cause complications during pregnancy and be involved in histamine intolerance. so far hdao has been characterized after isolation from either native sources or the heterologous production in insect cells. accessibility to human enzyme is limited and insect cells produce non-human glycosylation patterns that may alter its biochemical properties. we present the heterologous expression of hda ...201627063138
poly-pathway model, a novel approach to simulate multiple metabolic states by reaction network-based model - application to amino acid depletion in cho cell culture.mammalian cell lines are characterized by a complex and flexible metabolism. a single model that could describe the variations in metabolic behavior triggered by variations in the culture conditions would be a precious tool in bioprocess development. in this paper, we introduce an approach to generate a poly-pathway model and use it to simulate diverse metabolic states triggered in response to removal, reduction or doubling of amino acids in the culture medium of an antibody-producing cho cell l ...201627060554
polysorbates 20 and 80 degradation by group xv lysosomal phospholipase a2 isomer x1 in monoclonal antibody formulations.decreases in polysorbate (ps80) content were observed while evaluating the long-term storage stability of chinese hamster ovary-derived, purified monoclonal antibodies. it was determined that polysorbate had been enzymatically degraded; therefore, studies were performed to identify and characterize the protein(s) responsible. polysorbate degrading activity was enriched from chinese hamster ovary media leading to the identification of group xv lysosomal phospholipase a2 isomer x1 (lpla2) by shotg ...201627056628
cytotoxic and genotoxic evaluation of different synthetic amorphous silica nanomaterials in the v79 cell line.the nature of occupational risks and hazards in industries that produce or use synthetic amorphous silica (sas) nanoparticles is still under discussion. manufactured sas occur in amorphous form and can be divided into two main types according to the production process, namely, pyrogenic silica (powder) and precipitated silica (powder, gel or colloid). the physical and chemical properties of sas may vary in terms of particle size, surface area, agglomeration state or purity, and differences in th ...201625757481
in vitro and in vivo antitumor activity of a novel carbonyl ruthenium compound, the ct-[rucl(co)(dppb)(bipy)]pf-6[dppb=1,4-bis(diphenylphosphine)butane and bipy=2,2'-bipyridine].this study performed in vitro and in vivo biological assays of the ruthenium (ii) compound ct-[rucl(co)(dppb)(bipy)]pf6 (where, dppb=1,4-bis(diphenylphosphine)butane and bipy=2,2'-bipyridine). the cytotoxic activity of this compound was evaluated against different tumor cell lines (hela, human cervical adenocarcinoma; mcf7, human breast adenocarcinoma; mo59j, human glioblastoma; hepg2, hepatocellular carcinoma and b16f10, murine melanoma) and healthy cell line (v79, chinese hamster lung fibrobla ...201627613330
analytical performance and validation of a bioassay for thyroid-blocking antibodies.a cell-based bioassay for the measurement of thyroid blocking autoantibodies (tbab) has been recently reported. the analytical performance and validation of this bioassay is assessed and described.201626956921
data for amino acid alignment of japanese stingray melanocortin receptors with other gnathostome melanocortin receptor sequences, and the ligand selectivity of japanese stingray melanocortin receptors.this article contains structure and pharmacological characteristics of melanocortin receptors (mcrs) related to research published in "characterization of melanocortin receptors from stingray dasyatis akajei, a cartilaginous fish" (takahashi et al., 2016) [1]. the amino acid sequences of the stingray, d. akajei, mc1r, mc2r, mc3r, mc4r, and mc5r were aligned with the corresponding melanocortin receptor sequences from the elephant shark, callorhinchus milii, the dogfish, squalus acanthias, the gol ...201627408924
cell growth stimulating effect of ganoderma lucidum spores and their potential application for chinese hamster ovary k1 cell cultivation.in this work, water-soluble extracts of ganoderma lucidum spores (gls), a chinese medicinal herb that possesses cell growth stimulating function, were found to be an effective growth factor for chinese hamster ovary (cho) cell cultivation. the gls extract was prepared and supplemented to cho k1 cell culture media with various serum levels. our results obtained from both the static culture and the spinner-flask suspension culture showed that use of small-amount gls extract effectively promoted ce ...201626921102
the homologous recombination protein rad51d protects the genome from large deletions.homologous recombination (hr) is a dna double-strand break (dsb) repair pathway that protects the genome from chromosomal instability. rad51 mediator proteins (i.e. paralogs) are critical for efficient hr in mammalian cells. however, how hr-deficient cells process dsbs is not clear. here, we utilized a loss-of-function hr-reporter substrate to simultaneously monitor hr-mediated gene conversion and non-conservative mutation events. the assay is designed around a heteroallelic duplication of the a ...201627924006
protective effect of an isoflavone, tectorigenin, against oxidative stress-induced cell death via catalase activation.isoflavones are biologically active compounds that occur naturally in a variety of plants, with relatively high levels in soybean. tectorigenin, an isoflavone, protects against hydrogen peroxide (h2o2)-induced cell damage. however, the underlying mechanism is unknown.201628053960
peroxisome biogenesis and human peroxisome-deficiency disorders.peroxisome is a single-membrane-bounded ubiquitous organelle containing a hundred different enzymes that catalyze various metabolic pathways such as β-oxidation of very long-chain fatty acids and synthesis of plasmalogens. to investigate peroxisome biogenesis and human peroxisome biogenesis disorders (pbds) including zellweger syndrome, more than a dozen different complementation groups of chinese hamster ovary (cho) cell mutants impaired in peroxisome biogenesis are isolated as a model experime ...201627941306
site-1 protease is required for the generation of soluble (pro)renin receptor.the extracellular domain of the (pro)renin receptor [(p)rr] is cleaved to generate the soluble form of (p)rr [s(p)rr]. multiple clinical studies have revealed the association between serum/plasma s(p)rr levels and certain diseases, thereby suggesting a potential role for s(p)rr as a disease biomarker. here, we investigated whether site-1 protease (s1p) is responsible for cleaving (p)rr to generate s(p)rr. reduction of endogenous s1p with sirna attenuated s(p)rr generation in chinese hamster ovar ...201628013223
degranulation response in cytotoxic rat lymphocytes measured with a novel cd107a antibody.measuring degranulation through cd107a expression has become an advantageous tool for testing the functional capacity of cytotoxic cells. such functional studies have been hampered in the rat by the lack of a suitable anti-rat cd107a antibody. in this study, we report a novel hybridoma generated by immunizing armenian inbred hamsters with transfected chinese hamster ovary cells expressing cd107a. the sim1 clone exhibited specific reactivity with cd107a and measured degranulation from natural kil ...201628003815
plasma membrane organization and dynamics is probe and cell line dependent.the action and interaction of membrane receptor proteins take place within the plasma membrane. the plasma membrane, however, is not a passive matrix. it rather takes an active role and regulates receptor distribution and function by its composition and the interaction of its lipid components with embedded and surrounding proteins. furthermore, it is not a homogenous fluid but contains lipid and protein domains of various sizes and characteristic lifetimes which are important in regulating recep ...201627998689
intracellular cho cell metabolite profiling reveals steady-state dependent metabolic fingerprints in perfusion culture.perfusion cell culture processes allow the steady-state culture of mammalian cells at high viable cell density, which is beneficial for overall product yields and homogeneity of product quality in the manufacturing of therapeutic proteins. in this study, the extent of metabolic steady state and the change of the metabolite profile between different steady states of an industrial chinese hamster ovary (cho) cell line producing a monoclonal antibody (mab) was investigated in stirred tank perfusion ...201627997765
modeling cell survival and change in amount of dna during protracted irradiation.hyper-radiosensitivity (hrs) is a well-known bioresponse under low-dose or low-dose-rate exposures. although disorder of the dna repair function, non-targeted effects and accumulation of cells in g2 have been experimentally observed, the mechanism for inducing hrs by long-term irradiation is still unclear. on the basis of biological experiments and a theoretical study, we have shown that change in the amount of dna associated with accumulation of cells in g2 enhances radiosensitivity. to demonst ...201627974510
synthesis and characterization of injectable sulfonate-containing hydrogels.sulfonate-containing hydrogels are of particular interest because of their tunable mechanical and swelling properties, as well as their biological effects. polysulfonate copolymers were synthesized by reacting 2-acrylamido-2-methylpropanesulfonic acid (amps), acrylamide (am), and acrylic acid (aa). we found that the incorporation rate of sulfonate-containing monomer and the molecular weight of the copolymer were significantly enhanced by increasing the ionic strength of the solution. we introduc ...201627936721
transient and stable cho expression, purification and characterization of novel hetero-dimeric bispecific igg antibodies.igg bispecific antibodies (bsabs) represent one of the preferred formats for bispecific antibody therapeutics due to their native-like igg properties and their monovalent binding to each target. most reported studies utilized transient expression in hek293 cells to produce bsabs. however, the expression of biotherapeutic molecules using stable cho cell lines is commonly used for biopharmaceutical manufacturing. unfortunately, limited information is available in the scientific literature on the e ...201627977915
bioreactor productivity and media cost comparison for different intensified cell culture processes.process intensification in biomanufacturing has attracted a great deal of interest in recent years. manufacturing platform improvements leading to higher cell density and bioreactor productivity have been pursued. here we evaluated a variety of intensified mammalian cell culture processes for producing monoclonal antibodies. cell culture operational modes including fed-batch (normal seeding density or high seeding density with n-1 perfusion), perfusion, and concentrated fed-batch (cfb) were asse ...201627977910
generation of domain-specific monoclonal antibodies against human glutaredoxin3.human glutaredoxin3 (hglrx3), which encodes a 37.4 kda protein, possesses an n-terminal trx homology domain followed by two tandem repeats of grx domains. glrx3 is expressed in many tissues and plays important roles in iron metabolism, antioxidant effect, cell proliferation and development, regulation of immune reaction, and tumorigenesis. the mechanisms underlying the biological function of glrx3 are still not clear. to facilitate the functional research of glrx3, in this study, monoclonal anti ...201627923109
a carbon dioxide stripping model for mammalian cell culture in manufacturing scale bioreactors.control of carbon dioxide within the optimum range is important in mammalian bioprocesses at the manufacturing scale in order to ensure robust cell growth, high protein yields, and consistent quality attributes. the majority of bioprocess development work is done in laboratory bioreactors, in which carbon dioxide levels are more easily controlled. some challenges in carbon dioxide control can present themselves when cell culture processes are scaled up, because carbon dioxide accumulation is a c ...201627922179
cell compatibility of an eposimal vector mediated by the characteristic motifs of matrix attachment regions.the characteristic sequence of β-interferon matrix attachment regions (mars) can mediate transgene expression via episomal vectors in chinese hamster ovary (cho) cells. however, the host cells were from hamster ovaries, which are not suitable target cells for gene therapy. in this study, we aimed to evaluate the suitability of 12 different human cell lines as target cells for gene therapy. we transfected the cells with episomal vectors and obtained colonies stably expressing the vector products ...201627915978
potent mutagenicity of some non-planar tri- and tetrachlorinated biphenyls in mammalian cells, human cyp2e1 being a major activating enzyme.polychlorinated biphenyls (pcbs) have been classified as human carcinogens. mutagenicity of lower chlorinated biphenyls as well as activation of transcription factors by some other congeners may contribute to the carcinogenicity of pcbs. recently, we reported that human cyp2e1 activates mono- and dichlorobiphenyls to mutagens. however, mutagenicity of other pcbs and the involvement of other cyps remained unknown. in this study, chinese hamster v79-derived cell lines genetically engineered for ex ...201627913846
metachromatic leukodystrophy: biochemical characterization of two (p.307glu→lys, p.318trp→cys) arylsulfatase a mutations.metachromatic leukodystrophy (mld) is a lysosomal storage disease caused by arylsulfatase a (asa) deficiency. the hallmark of the disease is central and peripheral neurodegeneration. more than 200 mutations have been identified in arsa gene so far. some of these mutations were characterized. the aim of this study is to reinforce genotype-phenotype correlation and to understand the effect of mutations on the enzyme by biochemical characterization. two missense mutations (c.919g→a, p.307glu→lys an ...201627904824
jmv2894, a novel growth hormone secretagogue, accelerates body mass recovery in an experimental model of cachexia.oncologic patients subjected to chemotherapy frequently present aphagia, malnutrition, and cachexia. the purpose of this study was to investigate whether selected growth hormone secretagogues including hexarelin, jmv2894 and jmv2951 could antagonize body weight loss and wasting induced by cisplatin administration in rats. the three growth hormone secretagogues behaved as full agonists of the growth hormone secretagogues receptor both in terms of ability to stimulate calcium mobilization in chine ...201627896546
origanum majorana essential oil lacks mutagenic activity in the salmonella/microsome and micronucleus assays.the present study aimed to investigate the in vitro mutagenic activity of origanum majorana essential oil. the most abundant compounds identified by gc-ms were γ-terpinene (25.73%), α-terpinene (17.35%), terpinen-4-ol (17.24%), and sabinene (10.8%). mutagenicity was evaluated by the salmonella/microsome test using the preincubation procedure on ta98, ta97a, ta100, ta102, and ta1535 salmonella typhimurium strains, in the absence or in the presence of metabolic activation. cytotoxicity was detecte ...201627891531
a consensus genome-scale reconstruction of chinese hamster ovary cell metabolism.chinese hamster ovary (cho) cells dominate biotherapeutic protein production and are widely used in mammalian cell line engineering research. to elucidate metabolic bottlenecks in protein production and to guide cell engineering and bioprocess optimization, we reconstructed the metabolic pathways in cho and associated them with >1,700 genes in the cricetulus griseus genome. the genome-scale metabolic model based on this reconstruction, icho1766, and cell-line-specific models for cho-k1, cho-s, a ...201627883890
cho cells can make more protein.a new comprehensive genome-scale metabolic model of chinese hamster ovary cells identifies pathways for improving biopharmaceutical protein production.201627883886
novel role of er stress and autophagy in microcystin-lr induced apoptosis in chinese hamster ovary cells.microcystin-lr (mc-lr) is a ubiquitous peptide that exhibits strong reproductive toxicity, although the mechanistic basis for such toxicity remains largely unknown. the present study was conducted to investigate the mechanisms underlying the adverse effects of exposure to mc-lr in chinese hamster ovary (cho) cells. the results showed that mc-lr inhibited the in vitro proliferation of cho cells significantly, with an ic50 of 10 μm. moreover, mc-lr-treated cho cells revealed strong induction of ce ...201627877136
a nucleoside anticancer drug, 1-(3-c-ethynyl-β-d-ribo-pentofuranosyl)cytosine, induces depth-dependent enhancement of tumor cell death in spread-out bragg peak (sobp) of proton beam.the effect of 1-(3-c-ethynyl-β-d-ribo-pentofuranosyl)cytosine (ecyd) on proton-induced cell death was evaluated in human lung carcinoma cell line a549 and chinese hamster fibroblast cell line v79 to enhance relative biological effectiveness (rbe) within the spread-out bragg peak (sobp) of proton beams. treatment with ecyd significantly enhanced the proton-induced loss of clonogenicity and increased senescence at the center, but not at the distal edge of sobp. the p53-binding protein 1 foci forma ...201627875573
engineering translation in mammalian cell factories to increase protein yield: the unexpected use of long non-coding sineup rnas.mammalian cells are an indispensable tool for the production of recombinant proteins in contexts where function depends on post-translational modifications. among them, chinese hamster ovary (cho) cells are the primary factories for the production of therapeutic proteins, including monoclonal antibodies (mabs). to improve expression and stability, several methodologies have been adopted, including methods based on media formulation, selective pressure and cell- or vector engineering. this review ...201627872686
model-based investigation of intracellular processes determining antibody fc-glycosylation under mild hypothermia.despite the positive effects of mild hypothermic conditions on monoclonal antibody (mab) productivity (qmab ) during mammalian cell culture, the impact of reduced culture temperature on mab fc-glycosylation and the mechanism behind changes in the glycan composition are not fully established. the lack of knowledge about the regulation of dynamic intracellular processes under mild hypothermia restricts bioprocess optimization. to address this issue, a mathematical model that quantitatively describ ...201627869292
phenylpropanoids from cinnamon bark reduced β-amyloid production by the inhibition of β-secretase in chinese hamster ovarian cells stably expressing amyloid precursor protein.β-amyloid (aβ) is a substance of alzheimer disease (ad), which is generated via the amyloidogenic pathway from amyloid precursor protein (app) by β-secretase and γ-secretase. inhibition of aβ production is a potential therapeutic approach to ad. thus, we tested the hypothesis that cinnamon bark (cinnamomi cortex spissus), the dried bark of cinnamomum cassia blume (lauraceae), and its constituents are beneficial to ad. the methanol extract of cinnamon bark efficiently reduced aβ40 production in c ...201627865616
assessment of organic micropollutants occurrence in treated wastewater using heat shock protein 47 stress responses in chinese hamster ovary cells and gc/ms-based non-target screening.combining bioassays and analytical chemistry screening is a powerful approach to assess organic micropollutants which are the main contributors to toxic potential in complex mixtures of treated wastewater (tww). the aim of this study was to perform a comprehensive toxicity assessment of treated effluents using stress response bioassays and then to assess the occurrence of the organic micropollutants which were responsible for this biological response using gas chromatography coupled with a mass ...201627858797
genetic analyses of novel compound heterozygous hypodysfibrinogenemia, tsukuba i: fgg c.1129+62_65 del aata and fgg c.1299+4 del a.we found a novel hypodysfibrinogenemia designated tsukuba i caused by compound heterozygous nucleotide deletions with fgg c.1129+62_65 del aata and fgg c.1299+4 del a on different alleles. the former was deep in intron 8 of fgg (ivs-8 deletion) and the latter in exon 9 of fgg (ex-9 deletion), which is translated for the γ'-chain, but not the γa-chain. a western blot analysis of plasma fibrinogen from our patient revealed an aberrant γ-chain that migrated slightly faster than the normal bβ-chain.201627837696
functional effect of the mutations similar to the cleavage during platelet activation at integrin β3 cytoplasmic tail when expressed in mouse platelets.previous studies in chinese hamster ovary cells showed that truncational mutations of β3 at sites of f754 and y759 mimicking calpain cleavage regulate integrin signaling. the roles of the sequence from f754 to c-terminus and the conservative n756ity759 motif in platelet function have yet to be elaborated. mice expressing β3 with f754 and y759 truncations, or nity deletion (β3-δtnityrgt, β3-δrgt, or β3-δnity) were established through transplanting the homozygous β3-deficient mouse bone marrow cel ...201627851790
akt activation by evodiae fructus extract protects ovary against 4-vinylcyclohexene diepoxide-induced ovotoxicity.evodiae fructus (ef) is the dried, unripe fruit of evodia rutaecarpa benth., and one of the main components of traditional herbal prescriptions issued for the treatment of sterility caused by irregular menstruation in korea. however, scientific evidence regarding the efficacy and action mechanism of ef is lacking.201627769945
non-proton ligand-sensing domain of acid-sensing ion channel 3 is required for itch sensation.itch, the unpleasant sensation that evokes a desire to scratch, accompanies numerous skin and nervous system disorders. however, the molecular mechanisms of itch are unclear. acid-sensing ion channel 3 (asic3) is a sensor of acidic and primary inflammatory pain. the whole-cell patch clamp technique was used to determine the effect of chloroquine (cq) on asics currents in primary sensory neurons or the chinese hamster ovary cells transfected with rat asic1a or asic3. site-directed mutagenesis of ...201627770439
evaluating immunogenicity risk due to host cell protein impurities in antibody-based biotherapeutics.a potential risk factor for immunogenicity of a biotherapeutic is the low levels of host cell protein (hcp) impurities that remain in the product following the purification process. during process development, significant attention has been devoted to removing hcps due to their potential safety risk. samples from different purification steps of several monoclonal antibodies (mabs) purified by one type of platform were evaluated for their residual chinese hamster ovary (cho) cell-derived hcp cont ...201627450229
small molecule additive enhances cell uptake of 5-aminolevulinic acid and conversion to protoporphyrin ix.administration of exogenous 5-aminolevulinic acid (5-ala) to cancerous tissue leads to intracellular production of photoactive protoporphyrin ix, a biosynthetic process that enables photodynamic therapy and fluorescence-guided surgery of cancer. cell uptake of 5-ala is limited by its polar structure and there is a need for non-toxic chemical additives that can enhance its cell permeation. two zinc-bis(dipicolylamine) (znbdpa) compounds were evaluated for their ability to promote uptake of 5-ala ...201627722428
regulation of human cardiac kv1.5 channels by extracellular acidification.human kv1.5 channels (hkv1.5) conduct the ultra-rapid delayed rectifier potassium current (i kur), which plays an important role in action potential repolarization of atrial myocytes. the present study was undertaken to examine the effects of acidic ph on hkv1.5 wild-type (wt) and its pore mutant channels heterologously expressed in chinese hamster ovary (cho) cells using site-directed mutagenesis combined with whole-cell patch-clamp technique. both extracellular and intracellular acidifications ...201627796577
imiglucerase in the management of gaucher disease type 1: an evidence-based review of its place in therapy.gaucher disease is the first lysosomal disease to benefit from enzyme replacement therapy, thus serving as model for numerous other lysosomal diseases. alglucerase was the first glucocerebrosidase purified from placental extracts, and this was then replaced by imiglucerase - a chinese hamster ovary cell-derived glucocerebrosidase.201627790078
quaternary structures of opsin in live cells revealed by fret spectrometry.rhodopsin is a prototypical g-protein-coupled receptor (gpcr) that initiates phototransduction in the retina. the receptor consists of the apoprotein opsin covalently linked to the inverse agonist 11-cis retinal. rhodopsin and opsin have been shown to form oligomers within the outer segment disc membranes of rod photoreceptor cells. however, the physiological relevance of the observed oligomers has been questioned since observations were made on samples prepared from the retina at low temperatur ...201627623775
in vitro and in vivo pharmacological characterization of ssd114, a novel gabab positive allosteric modulator.positive allosteric modulators (pams) of the gabab receptor have emerged as a novel approach to the pharmacological manipulation of the gabab receptor, enhancing the effects of receptor agonists with few side effects. here, we identified n-cyclohexyl-4-methoxy-6-(4-(trifluoromethyl)phenyl)pyrimidin-2-amine (ssd114) as a new compound with activity as a gabab pam in in vitro and in vivo assays. ssd114 potentiated gaba-stimulated [(35)s]gtpγs binding to native gabab receptors, whereas it had no eff ...201627578262
correlations of in vitro assays for assessing cytotoxicity and biocompatibility of contact lens multipurpose solutions.to demonstrate correlations among in vitro assays used for assessing cytotoxicity of contact lens multipurpose solution (mps) and propose the use of multiple assays as a part of preclinical evaluation for mps biocompatibility assessment.201627749498
her2-targeted recombinant protein immuno-caspase-6 effectively induces apoptosis in her2-overexpressing gbm cells in vitro and in vivo.glioblastoma multiforme (gbm), which is associated with a high rate of morbidity and mortality, is among the most malignant and treatment-refractory neoplasms in human adults. as gbm is highly resistant to conventional therapies, immunotherapies are a promising treatment candidate. her2 is an attractive target for gbm immunotherapy, as its expression is highly associated with various types of gbm. we previously reported that a novel her2-targeted recombinant protein e23sfv-fdt-casp6 has an antit ...201627633091
a biotin-conjugated glutathione-responsive fret-based fluorescent probe with a ferrocenyl bodipy as the dark quencher.an efficient ferrocenyl bodipy based dark quencher has been developed and employed to construct a fret-based fluorescent probe that contains a biotin moiety as a potential directing ligand for cancer cells and a glutathione-cleavable disulfide linker connecting the quencher and a distyryl bodipy fluorophore. this molecular probe is deactivated in the native form through fret followed by intramolecular charge transfer due to the ferrocenyl unit. however, upon interaction with glutathione in phosp ...201627774548
a simple method to determine igg light chain to heavy chain polypeptide ratios expressed by cho cells.to establish a high-throughput method for determination of antibodies intra- and extracellular light chain (lc) to heavy chain (hc) polypeptide ratio as screening parameter during cell line development.201627613153
investigation on tissue specific effects of pro-apoptotic micro rnas revealed mir-147b as a potential biomarker in ovarian cancer prognosis.the development and progression of cancer can be ascribed to imbalances in gene regulation leading to aberrant cellular behavior. the loss of micro rnas (mirnas) exhibiting tumor-suppressive function has been demonstrated to be often causative for uncontrolled cell proliferation, migration or tissue infiltration. the installation of de novo tumor suppressive function by using pro-apoptotic mirnas might be a promising therapeutic approach. in addition, there is a great demand for novel biomarkers ...201627821806
osmotic effects induced by pore-forming agent nystatin: from lipid vesicles to the cell.the responses of chinese hamster ovary epithelial cells, caused by the pore-forming agent nystatin, were investigated using brightfield and fluorescence microscopy. different phenomena, i.e., the detachment of cells, the formation of blebs, the occurrence of "cell-vesicles" and cell ruptures, were observed. these phenomena were compared to those discovered in giant lipid vesicles. a theoretical model, based on the osmotic effects that occur due to the size-discriminating nystatin transmembrane p ...201627788169
flexible nanopillars to regulate cell adhesion and movement.flexible polymer nanopillar substrates were used to systematically demonstrate cell alignment and migration guided by the directional formation of focal adhesions. the polymer nanopillar substrates were constructed to various height specifications to provide an extensive variation of flexibility; a rectangular arrangement created spatial confinement between adjacent nanopillars, providing less spacing in the horizontal and vertical directions. three polymer nanopillar substrates with the diamete ...201627775920
a comparison of orbitally-shaken and stirred-tank bioreactors: ph modulation and bioreactor type affect cho cell growth and protein glycosylation.orbitally shaken bioreactors (osrs) support the suspension cultivation of animal cells at volumetric scales up to 200 l and are a potential alternative to stirred-tank bioreactors (strs) due to their rapid and homogeneous mixing and high oxygen transfer rate. in this study, a chinese hamster ovary cell line producing a recombinant antibody was cultivated in a 5 l osr and a 3 l str, both operated with or without ph control. effects of bioreactor type and ph control on cell growth and metabolism a ...201627453130
multigene expression in stable cho cell pools generated with the piggybac transposon system.heterogenous populations of recombinant cells (cell pools) stably expressing 1-4 transgenes were generated from chinese hamster overy (cho) cells with the piggybac (pb) transposon system. the cell pools produced different combinations of three model proteins-enhanced green fluorescent protein (egfp), secreted alkaline phosphatase (seap), and a monoclonal igg1 antibody. each transgene was present on a separate pb donor plasmid with either the same or a different selection gene. in both cases, we ...201627302570
cholesterol-lowering effect of octaarginine-appended β-cyclodextrin in npc1-trap-cho cells.niemann-pick disease type c (npc) is an autosomal recessive lysosomal storage disorder, which is an inherited disease characterized by the accumulation of unesterified cholesterol in endolysosomes. recently, 2-hydroxypropyl-β-cyclodextrin (hp-β-cyd) has been used for the treatment of npc, and ameliorated a hepatosplenomegaly in the patients. however, to obtain the treatment efficacy, a high dose of hp-β-cyd was necessary. therefore, the decrease in dose by using active intracellular delivery sys ...201627601082
evaluating the use of a cpg free promoter for long-term recombinant protein expression stability in chinese hamster ovary cells.methylated cpg dinucleotides in promoters are associated with the loss of gene expression in recombinant chinese hamster ovary (cho) cells during large-scale commercial manufacturing. we evaluated a promoter devoid of cpg dinucleotides, cpgfree, in parallel with a similar cpg containing promoter, cpgrich, for their ability to maintain the expression of recombinant enhanced green fluorescent protein (egfp) after 8 weeks of culturing.201627756290
antinociceptive effects of the 6-o-sulfate ester of morphine in normal and diabetic rats: comparative role of mu- and delta-opioid receptors.this study determined the antinociceptive effects of morphine and morphine-6-o-sulfate (m6s) in both normal and diabetic rats, and evaluated the comparative role of mu-opioid receptors (mu-ors) and delta-opioid receptors (delta-ors) in the antinociceptive action of these opioids. in vitro characterization of mu-or and delta-or-mediated signaling by m6s and morphine in stably transfected chinese hamster ovary (cho-k1) cells showed that m6s exhibited a 6-fold higher affinity for delta-ors and modu ...201627637375
inhibition of the ubiquitin ligase activity improves the production of biologically active fusion protein hsa-hgf in chinese hamster ovary cells.hepatocyte growth factor (hgf) is a potent multi-functional protein that stimulates proliferation, survival, motility, scattering and differentiation during growth and development, and has been considered to be a potential therapeutic agent for the treatment of a number of intractable diseases. the aim of this study was to enhance the expression of recombinant fusion protein hsa-hgf (r494e) in cho cells by inhibiting the intracellular ubiquitin ligase activity. the high stable expression sub-clo ...201627753513
mitochondrial genome of cricetulus migratorius (rodentia: cricetidae): insights into the characteristics of the mitochondrial genome and the phylogenetic relationships of cricetulus species.cricetulus migratorius is widely distributed in the northwestern arid regions of china. here, the complete mitochondrial genome (mitogenome) of c. migratorius is reported, to our knowledge, for the first time. it was found to be 16,246bp in length, including 13 protein-coding genes (pcgs), 2 rrna genes, 22 trna genes, and one control region, and showed characteristics typical of the vertebrate mitogenome. comparative analyses of mitogenomes of cricetulus species showed that hamster mitogenomes h ...201627717890
structural and antitrypanosomal data of different carbasones of piperitone.this article reports data on four carbazones of piperitone: semicarbazone 1, thiosemicarbazone 2, 4-phenyl semicarbazone 3 and 4-phenyl thiosemicarbazone 4 prepared directly in situ from essential oil of cymbopogon schoenantus, whose gc-fid and gc-ms analysis revealed piperitone as major component (68.20%). the structures of hemi-synthesized compounds were confirmed by high throughput ir, ms, (1)h and (13)c nmr based spectrometric analysis. their antiparasitic activities were evaluated in vitro ...201627900358
a new family of insect muscarinic acetylcholine receptors.most currently used insecticides are neurotoxic chemicals that target a limited number of sites and insect cholinergic neurotransmission is the major target. a potential target for insecticide development is the muscarinic acetylcholine receptor (machr), which is a metabotropic g-protein-coupled receptor. insects have a- and b-type machrs and the five mammalian machrs are close to the a-type. we isolated a cdna (cg12796) from the fruit fly, drosophila melanogaster. after heterologous expression ...201627003873
development of betulinic acid as an agonist of tgr5 receptor using a new in vitro assay.g-protein-coupled bile acid receptor 1, also known as tgr5 is known to be involved in glucose homeostasis. in animal models, treatment with a tgr5 agonist induces incretin secretion to reduce hyperglycemia. betulinic acid, a triterpenoid present in the leaves of white birch, has been introduced as a selective tgr5 agonist. however, direct activation of tgr5 by betulinic acid has not yet been reported.201627578964
serum albumin attenuates the open-channel blocking effects of propofol on the human kv1.5 channel.the intravenous anesthetic propofol modulates various ion channel functions. it is generally accepted that approximately 98% of propofol binds to blood constituents and that the free (unbound) drug preferentially affects target proteins including ion channels. however, modulatory effects of propofol on ion channels have not been previously explored in the presence of serum albumin. this study was designed to investigate the effects of serum albumin on the blocking action of propofol on the human ...201627164421
effects of isoleucine deficiency on nucleic acid and protein metabolism in cultured chinese hamster cells. continued ribonucleic acid and protein synthesis in the absence of deoxyribonucleic acid synthesis. 20165061874
effect of circadian rhythm on the radiation response of the chinese hamster (cricetulus griseus). 20165030088
intermittent dna synthesis and periodic expression of enzyme activity in the cell cycle of wi-38.synchronous cultures of wi-38 were obtained using an automated system for detachment and partitioning of mitotic cells which operates without the use of inhibitors, altered medium, or lowered temperatures. the generation time in synchronous wi-38 is 19.5 h and the duration of s phase when determined from the percentage of labeled metaphase cells or nuclei is 12 h. dna replication in wi-38 occurs in three temporally distinct and rapid bursts separated by intervals of greatly reduced synthesis wit ...20164795861
studies on induced aberrations in diplochromosomes of chinese hamster cells. 20164796813
temporal variation of adenine ribonucleotides during the cell cycle of chinese hamster fibroblasts in culture. 20164855877
[anatomical studies on the urinary tract and genitalia of the chinese dwarf hamster (cricetulus griseus)]. 20165036384
n-ethylmaleimide and the cyclic response to x-rays of synchronous chinese hamster cells. 20164736799
islet and beta cell volumes in offspring of severely diabetic (ketotic) chinese hamsters. 20164914657
chromosomal aberrations induced by caffeine in endoreduplicated chinese hamster cells. 20164822595
clones of chinese hamster cells cultivated in vitro not permanently resistant to azaguanine. 20164843976
antioxidant and dna repair stimulating effect of extracts from transformed and normal roots of rhaponticum carthamoides against induced oxidative stress and dna damage in cho cells.rhaponticum carthamoides has a long tradition of use in siberian folk medicine. the roots and rhizomes of this species are used in various dietary supplements or nutraceutical preparations to increase energy level or eliminate physical weakness. this is the first report to reveal the protective and dna repair stimulating abilities of r. carthamoides root extracts in chinese hamster ovary (cho) cells exposed to an oxidative agent. both transformed root extract (tr extract) and extract of soil-gro ...201627034736
ultrastructural changes in a-cells exposed to diabetic hyperglycaemia. observations made on pancreas of chinese hamsters. 20164914660
the retinal vasculature of the chinese hamster: a preliminary study. 20164914659
high-efficiency secretory expression of human neutrophil gelatinase-associated lipocalin from mammalian cell lines with human serum albumin signal peptide.human neutrophil gelatinase associated lipocalin (ngal) is a secretory glycoprotein initially isolated from neutrophils. it is thought to be involved in the incidence and development of immunological diseases and cancers. urinary and serum levels of ngal have been investigated as a new biomarker of acute kidney injury (aki), for an earlier and more accurate detection method than with creatinine level. however, expressing high-quality recombinant ngal is difficult both in escherichia coli and mam ...201626518367
bioisosteric ferrocenyl-containing quinolines with antiplasmodial and antitrichomonal properties.bioisosteric ferrocenyl-containing quinolines and ferrocenylamines containing organosilanes and their carbon analogues, were prepared and fully characterised. the molecular structures of two ferrocenyl-containing quinolines, determined using single-crystal x-ray diffraction, revealed that the compounds crystallise in a folded conformation. the compounds were screened for their antiplasmodial activity against the chloroquine-sensitive (nf54) and cq-resistant (dd2) strains of p. falciparum, as wel ...201627858010
biophysical studies on calcium and carbohydrate binding to carbohydrate recognition domain of gal/galnac lectin from entamoeba histolytica: insights into host cell adhesion.entamoeba histolytica, an enteric parasite expresses a gal/galnac-specific lectin that contributes to its virulence by establishing adhesion to host cell. in this study, carbohydrate recognition domain of hgl (ehcrd) was purified and biophysical studies were conducted to understand the thermodynamic basis of its binding to carbohydrate and ca(++) here, we show that carbohydrate recognition domain (crd) of the lectin binds to calcium through dpn motif. to decipher the role of calcium in carbohydr ...201627008865
general and genetic toxicology of enzyme-treated ginseng extract: toxicology of ginseng rh2.ginseng rh2+ is enzyme-treated ginseng extract containing high amounts of converted ginsenosides, such as compound k, rh2, rg3, which have potent anticancer activity. we conducted general and genetic toxicity tests to evaluate the safety of ginseng rh2+.201627695630
calibrated complex impedance of cho cells and e. coli bacteria at ghz frequencies using scanning microwave microscopy.the application of scanning microwave microscopy (smm) to extract calibrated electrical properties of cells and bacteria in air is presented. from the s 11 images, after calibration, complex impedance and admittance images of chinese hamster ovary cells and e. coli bacteria deposited on a silicon substrate have been obtained. the broadband capabilities of smm have been used to characterize the bio-samples between 2 ghz and 20 ghz. the resulting calibrated cell and bacteria admittance at 19 ghz w ...201626895571
multi-omics data driven analysis establishes reference codon biases for synthetic gene design in microbial and mammalian cells.in this study, we analyzed multi-omics data and subsets thereof to establish reference codon usage biases for codon optimization in synthetic gene design. specifically, publicly available genomic, transcriptomic, proteomic and translatomic data for microbial and mammalian expression hosts, escherichia coli, saccharomyces cerevisiae, pichia pastoris and chinese hamster ovary (cho) cells, were compiled to derive their individual codon and codon pair frequencies. then, host dependent and -omics spe ...201626850284
[study of in vitro expression of human platelet itgb3 gene nonsense mutation c.1476g>a].to explore the function of a novel nonsense mutation c.1476g>a of itgb3 gene using an in vitro expression system.201626829726
fast filtration of bacterial or mammalian suspension cell cultures for optimal metabolomics results.the metabolome offers real time detection of the adaptive, multi-parametric response of the organisms to environmental changes, pathophysiological stimuli or genetic modifications and thus rationalizes the optimization of cell cultures in bioprocessing. in bioprocessing the measurement of physiological intracellular metabolite levels is imperative for successful applications. however, a sampling method applicable to all cell types with little to no validation effort which simultaneously offers h ...201627438065
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