| cytotoxic and genotoxic effects of e-liquids and their potential associations with nicotine, menthol and phthalate esters. | in this study, we determined dna damage and chromosome breakage (indicators of genotoxicity) and cell viability (an indicator of cytotoxicity) in human lymphoblastoid tk6 and chinese hamster ovary (cho) cells treated with 33 e-liquids using in vitro single cell gel (comet), micronucleus (mn), and trypan blue assays, respectively. we also measured the contents of nicotine, five phthalate esters, and dl-menthol in the e-liquids to examine their effects on dna damage, chromosome breakage, and cell ... | 2020 | 32058129 |
| eco-friendly synthesis of glutathione-capped cdte/cdse/znse core/double shell quantum dots: their cytotoxicity and genotoxicity effects on chinese hamster ovary cells. | in this work, we report green one-pot synthesis, cytotoxicity and genotoxicity of glutathione-capped cdte/cdse/znse heterostructured quantum dots (qds) using a label-free xcelligence rtca system as well as the cytokinesis blocked micronucleus assay. the as-synthesised nanocrystals displayed good optical properties and were spherical in shape with an average particle diameter of 5.9 ± 1.13 nm. the intracellular uptake study showed that most of the as-synthesised glutathione stabilized qds penetra ... | 2019 | 32055393 |
| codon and amino acid content are associated with mrna stability in mammalian cells. | messenger rna (mrna) degradation plays a critical role in regulating transcript levels in the cell and is a major control point for modulating gene expression. in yeast and other model organisms, codon identity is a powerful determinant of transcript stability, contributing broadly to impact half-lives. general principles governing mrna stability are poorly understood in mammalian systems. importantly, however, the degradation machinery is highly conserved, thus it seems logical that mammalian t ... | 2020 | 32053646 |
| single-step synthesis and in vitro anti-mycobacterial activity of novel nitrofurantoin analogues. | the emergence of drug-resistant tuberculosis (dr-tb) as well as the requirement for long, expensive and toxic drug regimens impede efforts to control and eliminate tb. therefore, there's a need for effective and affordable anti-mycobacterial agents which can shorten the duration of therapy and are active against mycobacterium tuberculosis (mtb) in both active and latent phases. nitrofurantoin (nft) is a hypoxic agent with activity against a myriad of anaerobic pathogens and, like the first-line ... | 2020 | 32044516 |
| profiles of plant core-fucosylated n-glycans of acid alpha-glucosidases produced in transgenic rice cell suspension cultures treated with eight different conditions. | recombinant human acid alpha-glucosidase (rhgaa) from chinese hamster ovary cells is the only approved treatment for patients with pompe disease. in this study, rhgaas were produced in transgenic rice cell suspension cultures under eight different conditions; untreated, 5 μm of 2-fluoro-l-fucose (2-ff), 50 μm of 2-ff, 100 μm of 2-ff, 100 μm of 2-ff + 0.5% pluronic f-68 (pf-68), 100 μm of 2-ff + 0.05% tween 20 (tw 20), 0.5% pf-68, and 0.05% tw 20. the n-glycans of eight rhgaas were analyzed using ... | 2020 | 32044029 |
| c1q/tnf-related protein 6 is a pattern recognition molecule that recruits collectin-11 from the complement system to ligands. | c1q/tnf-related protein (ctrp) 6 is a member of the ctrp protein family associated with the regulation of cellular and endocrine processes. ctrp6 contains collagen and globular structures, resembling the pattern recognition molecules (prms) of the classical and lectin complement pathways. we expressed human ctrp6 in chinese hamster ovary cells and investigated the binding to different putative ligands (acetylated bsa [acbsa], zymosan, mannan, and lps from escherichia coli and salmonella as well ... | 2020 | 32041782 |
| engineering mammalian cells to produce plant-specific n-glycosylation on proteins. | protein n-glycosylation is an essential and highly conserved posttranslational modification found in all eukaryotic cells. yeast, plants and mammalian cells, however, produce n-glycans with distinct structural features. these species-specific features not only pose challenges in selecting host cells for production of recombinant therapeutics for human medical use but also provide opportunities to explore and utilize species-specific glycosylation in design of vaccines. here, we used reverse cros ... | 2020 | 32039452 |
| the epilepsy of infancy with migrating focal seizures: identification of de novo mutations of the kcnt2 gene that exert inhibitory effects on the corresponding heteromeric kna1.1/kna1.2 potassium channel. | the epilepsy of infancy with migrating focal seizures (eimfs; previously called malignant migrating partial seizures of infancy) are early-onset epileptic encephalopathies (eoee) that associate multifocal ictal discharges and profound psychomotor retardation. eimfs have a genetic origin and are mostly caused by de novo mutations in the kcnt1 gene, and much more rarely in the kcnt2 gene. kcnt1 and kcnt2 respectively encode the kna1.1 (slack) and kna1.2 (slick) subunits of the sodium-dependent vol ... | 2020 | 32038177 |
| inhibition of cholesteryl ester synthesis by polyacetylenes from atractylodes rhizome. | using activity guided purification, four known compounds, sesquiterpene atractylenolide iii (1), and the polyacetylenes 14-acetoxy-12-senecioyloxytetradeca-2e,8e,10e-trien-4,6-diyn-1-ol (2), 14-acetoxy-12-α-methylbutyl-2e,8e,10e-trien-4,6-diyn-1-ol (3), and 14-acetoxy-12-β -methylbutyl-2e,8e,10e-trien-4,6-diyn-1-ol (4), were isolated from a traditional herbal medicine, atractylodes rhizome. structurally similar 3 and 4 (3/4 mixture) were obtained as a mixture. in intact chinese hamster ovary (ch ... | 2020 | 32035699 |
| protective effects of compounds from cimicifuga dahurica against amyloid beta production in vitro and scopolamine-induced memory impairment in vivo. | cimicifuga dahurica has traditionally been used as an antipyretic, analgesic, and anti-inflammatory agent and as a treatment for uterine and anal prolapse. this study has investigated the potential beneficial effects of this medicinal plant and its components on alzheimer's disease (ad) with a focus on amyloid beta (aβ) production and scopolamine-induced memory impairment in mice. an ethanol extract from c. dahurica roots decreased aβ production in app-cho cells [chinese hamster ovarian (cho) ce ... | 2020 | 32031796 |
| glucose monitoring in cell culture with online ultrasound-assisted near-infrared spectroscopy. | near-infrared (nir) spectroscopic analysis for suspensions often requires a pretreatment by dilution or filtering of suspended solids to maintain the sensitivity of the measurements. an online ultrasound-assisted spectroscopy (uas) unit enabling pretreatment-free and noncontact analysis for bioprocessing is proposed and evaluated with a model suspension containing 3-μm-diameter polystyrene latex particles (psls) with the density of 5.1 × 108 counts/ml and chinese hamster ovary (cho) cell culture ... | 2020 | 32023047 |
| differential effects of 2-hydroxypropyl-cyclodextrins on lipid accumulation in npc1-null cells. | niemann-pick disease type c (npc) is an autosomal recessive disorder characterized by abnormal accumulation of free cholesterol and sphingolipids in lysosomes. the iminosugar miglustat, which inhibits hexosylceramide synthesis, is used for npc treatment, and 2-hydroxypropyl-β-cyclodextrin (hp-β-cd), a cyclic oligosaccharide derivative, is being developed to treat npc. moreover, therapeutic potential of 2-hydroxypropyl-γ-cyclodextrin (hp-γ-cd) was shown in npc models, although its mechanism of ac ... | 2020 | 32019132 |
| 3-quinuclidinyl-α-methoxydiphenylacetate: a multi-targeted ligand with antimuscarinic and antinicotinic effects designed for the treatment of anticholinesterase poisoning. | racemic 3-quinuclidinyl-α-methoxydiphenylacetate (mb266) was synthesised. its activity at muscarinic acetylcholine receptors (machrs), and muscle and neuronal nicotinic acetylcholine receptors (nachrs), was compared to that of atropine and racemic 3-quinucidinyl benzilate (qnb) using a functional assay based on agonist-induced elevation of intracellular calcium ion concentration in cn21, chinese hamster ovary (cho) and shsy5y human cell lines. mb266 acted as an antagonist at acetylcholine recept ... | 2020 | 32017982 |
| analysis of chemometric models applied to raman spectroscopy for monitoring key metabolites of cell culture. | the food and drug administration (fda) initiative of process analytical technology (pat) encourages the monitoring of biopharmaceutical manufacturing processes by innovative solutions. raman spectroscopy and the chemometric modeling tool partial least squares (pls) have been applied to this aim for monitoring cell culture process variables. this study compares the chemometric modeling methods of support vector machine radial (svmr), random forests (rf), and cubist to the commonly used linear pls ... | 2020 | 32012476 |
| feeding tricarboxylic acid cycle intermediates improves lactate consumption and antibody production in chinese hamster ovary cell cultures. | media components play an important role in modulating cell metabolism and improving product titer in mammalian cell cultures. to sustain cell productivity, highly active oxidative metabolism is desired. here we explored the effect of tricarboxylic acid (tca) cycle intermediates supplementation on lactate metabolism and productivity in chinese hamster ovary fed-batch cultures. direct addition of 5 mm alpha-ketoglutarate (α-kg), malic acid, or succinic acid in the basal medium did not have any sig ... | 2020 | 32012447 |
| structure-function relationships of the soluble form of the antiaging protein klotho have therapeutic implications for managing kidney disease. | the fortuitously discovered antiaging membrane protein αklotho (klotho) is highly expressed in the kidney, and deletion of the klotho gene in mice causes a phenotype strikingly similar to that of chronic kidney disease (ckd). klotho functions as a co-receptor for fibroblast growth factor 23 (fgf23) signaling, whereas its shed extracellular domain, soluble klotho (sklotho), carrying glycosidase activity, is a humoral factor that regulates renal health. low sklotho in ckd is associated with diseas ... | 2020 | 32005658 |
| anti-d monoclonal antibodies from 23 human and rodent cell lines display diverse igg fc-glycosylation profiles that determine their clinical efficacy. | anti-d immunoglobulin (anti-d ig) prophylaxis prevents haemolytic disease of the fetus and newborn. monoclonal igg anti-ds (mab-ds) would enable unlimited supplies but have differed in efficacy in fcγriiia-mediated adcc assays and clinical trials. structural variations of the oligosaccharide chains of mab-ds are hypothesised to be responsible. quantitative data on 12 fc-glycosylation features of 23 mab-ds (12 clones, 5 produced from multiple cell lines) and one blood donor-derived anti-d ig were ... | 2020 | 32001734 |
| the gβ1 and gβ3 subunits differentially regulate rat vascular kv7 channels. | within the vasculature kv7 channels are key regulators of basal tone and contribute to a variety of receptor mediated vasorelaxants. the kv7.4 isoform, abundant within the vasculature, is key to these processes and was recently shown to have an obligatory requirement of g-protein βγ subunits for its voltage dependent activity. there is an increasing appreciation that with 5 gβ subunits and 12 gγ subunits described in mammalian cells that different gβ x γ x combinations can confer selectivity in ... | 2019 | 31992990 |
| inhibition of endogenous mir-23a/mir-377 in cho cells enhances difficult-to-express recombinant lysosomal sulfatase activity. | difficult-to-express (dte) recombinant proteins such as multi-specific proteins, dte monoclonal antibodies, and lysosomal enzymes have seen difficulties in manufacturability using chinese hamster ovary (cho) cells or other mammalian cells as production platforms. cho cells are preferably used for recombinant protein production for their ability to secrete human-like recombinant proteins with posttranslational modification, resistance to viral infection, and familiarity with drug regulators. howe ... | 2020 | 31990124 |
| cytotoxic and genotoxic assessments of 2,4-dichlorophenoxyacetic acid (2,4-d) in in vitro mammalian cells. | a combined approach employing alkaline single cell gel electrophoresis (scge) and cytokinesis-blocked micronucleus (mns) cytome bioassays was adopted to assess the deleterious properties of the auxinic 2,4-dichlorophenoxyacetic acid (2,4-d) and its microparticulated low volatility product dedalo elite (30% a.i.) on chinese hamster ovary (cho-k1) cells. cytotoxicity was estimated by neutral red uptake (nru), succinic dehydrogenase activity (mtt) and apoptosis assessment. both compounds were assay ... | 2020 | 31987841 |
| recombinant β-glucocerebrosidase specific immunoaffinity ligands selected from phage-displayed combinatorial scfv libraries. | antibodies specific to β-glucocerebrosidase were selected from phage displayed naïve scfv libraries. biopannings were performed against recombinant human protein β-glucocerebrosidase immobilized on polystyrene surface, specific phages were eluted with 50% ethylene glycol in citrate buffer (ph 6.0). several specific binders were discovered and converted to full-size higg1 antibodies leading to highly stable binders with dissociation constants (kd) in the range 10-40 nm. the antibodies were used f ... | 2020 | 31981620 |
| an efficient exogenous gene insertion site in cho cells with high transcription level to enhance aid-induced mutation. | antibodies have been extensively used for the purpose of scientific research, clinical diagnosis, and therapy. combination of in vitro somatic hypermutation and mammalian cell surface display has been an efficient technology for antibody or other proteins optimization, in which the efficiency of activation-induced cytidine deaminase (aid) mutations in genes is one of the most important key factors. gene transcriptional level has been found to be positively proportional to aid-induced mutation fr ... | 2020 | 31975519 |
| maximizing antibody production in a targeted integration host by optimization of subunit gene dosage and position. | historically, therapeutic protein production in chinese hamster ovary (cho) cells has been accomplished by random integration (ri) of expression plasmids into the host cell genome. more recently, the development of targeted integration (ti) host cells has allowed for recombination of plasmid dna into a predetermined genomic locus, eliminating one contributor to clone-to-clone variability. in this study, a ti host capable of simultaneously integrating two plasmids at the same genomic site was use ... | 2020 | 31965756 |
| macrophages show higher levels of engulfment after disruption of cis interactions between cd47 and the checkpoint receptor sirpα. | the macrophage checkpoint receptor sirpα signals against phagocytosis by binding cd47 expressed on all cells - including macrophages. here, we found that inhibiting cis interactions between sirpα and cd47 on the same macrophage increased engulfment ('eating') by approximately the same level as inhibiting trans interactions. antibody blockade of cd47, as pursued in clinical trials against cancer, was applied separately to human-derived macrophages and to red blood cell (rbc) targets for phagocyto ... | 2020 | 31964705 |
| cell electropermeabilisation enhancement by non-thermal-plasma-treated pbs. | the effectiveness of electrochemotherapy (ect) in local eradication of tumours in human and veterinary medicine has been proven. ect consists of increasing the uptake of cytotoxic drugs by means of pulsed electric fields (pefs) that transiently permeabilise the cell membrane. still, this tumour treatment includes some drawbacks that are linked to the characteristics of the intense electric pulses (eps) used. meanwhile, the emerging field of cancer therapies that are based on the application of n ... | 2020 | 31963132 |
| defining lncrnas correlated with cho cell growth and igg productivity by rna-seq. | how the long non-coding rna (lncrna) genome in recombinant protein producing chinese hamster ovary (cho) cell lines relates to phenotype is not well described. we therefore defined the cho cell lncrna transcriptome from cells grown in controlled miniature bioreactors under fed-batch conditions using rna-seq to identify lncrnas and how the expression of these changes throughout growth and between igg producers. we identify lncrnas including adapt15, linked to er stress, gas5, linked to mtor signa ... | 2020 | 31962234 |
| heterozygous variant fibrinogen γa289v (kanazawa iii) was confirmed as hypodysfibrinogenemia by plasma and recombinant fibrinogens. | congenital fibrinogen disorders are classified as afibrinogenemia, hypofibrinogenemia, dysfibrinogenemia, and hypodysfibrinogenemia. however, difficulties are associated with discriminating between dysfibrinogenemia, hypofibrinogenemia, and hypodysfibrinogenemia using routine analyses. we previously reported a heterozygous variant fibrinogen (γa289v; kanazawa iii) as hypodysfibrinogenemia; however, the same variant had previously been described as hypofibrinogenemia. to clarify the production of ... | 2020 | 31957968 |
| influence of cytoskeleton organization on recombinant protein expression by cho cells. | in this study, we assessed the importance of cytoskeleton organization in the mammalian cells used to produce therapeutic proteins. two cytoskeletal genes, actin alpha cardiac muscle 1 (actc1) and a guanosine triphosphate gtpase-activating protein (tagap), were found to be upregulated in highly productive therapeutic protein-expressing chinese hamster ovary (cho) cells selected by the deprivation of vitamin b5. we report here that the overexpression of the actc1 protein was able to improve signi ... | 2020 | 31956990 |
| overexpression of transcription factor foxa1 and target genes remediate therapeutic protein production bottlenecks in chinese hamster ovary cells. | despite extensive research conducted to increase protein production from chinese hamster ovary (cho) cells, cellular bottlenecks often remain, hindering high yields. in this study, a transcriptomic analysis led to the identification of 32 genes that are consistently upregulated in high producer clones and thus might mediate high productivity. candidate genes were associated with functions such as signaling, protein folding, cytoskeleton organization, and cell survival. we focused on two engineer ... | 2020 | 31956982 |
| gene disruption of ribosomal protein l5 (rpl5) decreased the sensitivity of cho-k1 cells to uncoupler carbonylcyanide-3-chlorophenylhydrazone. | protonophoric uncoupler carbonylcyanide-3-chlorophenylhydrazone (cccp) decreases the proton motive force (δp) of the mitochondrial inner membrane and results in inhibition of oxidative phosphorylation. in this study, a cccp-resistant clone was isolated from a random gene trap insertional mutant library of chinese hamster ovary (cho)-k1 cells which was constructed by infecting a retrovirus vector, rosaβgeo. although we expected the isolation of the mutants defective in nuclear genes responsible f ... | 2019 | 31956228 |
| lipid catabolism in starved yak is inhibited by intravenous infusion of β-hydroxybutyrate. | lipid is the chief energy source for starved animals. β-hydroxybutyrate (bhba) is the main ketone body produced by lipid decomposition. in chinese hamster ovary (cho) cell experiment, it was found that bhba could be used not only as an energy substance, but also as a ligand of gpr109a for regulating lipid metabolism. however, whether bhba can regulate lipid metabolism of yaks, and its effective concentration and signal pathway are not clear. this study investigated the effects and mechanism of s ... | 2020 | 31952136 |
| electron scattering in conventional cell flask experiments and dose distribution dependency. | electron beam therapy (ebt) is commonly used for treating superficial and subdermal tumors. previous cellular radiosensitivity research using ebt may be underestimating the contribution from flask wall scattering and the corresponding dose distribution. single cell suspensions of chinese hamster ovary (cho) cells were plated on flasks and irradiated with 3, 4, 7, 9, and 18 mev energy electron beams from two different institutions, and the spatial locations of surviving colonies were recorded. ga ... | 2020 | 31949176 |
| correction of the aprt gene using repair-polypurine reverse hoogsteen hairpins in mammalian cells. | in this study, we describe the correction of single-point mutations in mammalian cells by repair-polypurine reverse hoogsteen hairpins (repair-pprhs). these molecules consist of (1) a pprh hairpin core that binds to a polypyrimidine target sequence in the double-stranded dna (dsdna), producing a triplex structure, and (2) an extension sequence homologous to the dna sequence to be repaired but containing the wild-type nucleotide instead of the mutation and acting as a donor dna to correct the mut ... | 2020 | 31945727 |
| photoacoustic effect applied on model membranes and living cells: direct observation with multiphoton excitation microscopy and long-term viability analysis. | the photoacoustic effect is generated when a variable light interacts with a strongly light-absorbing material. in water, it may produce hot bubbles and shock waves that could affect the integrity of nearby cellular membranes, opening transient pores (photoporation). in this study, we have evaluated the effect of pulsed laser-irradiated carbon nanoparticles (cnp) on model membranes and on chinese hamster ovary (cho) cells. fluorescence lifetime measurements of calcein-loaded liposomes support th ... | 2020 | 31941922 |
| effective temperature shift strategy development and scale confirmation for simultaneous optimization of protein productivity and quality in chinese hamster ovary cells. | temperature shifts to lower culture temperatures are frequently employed in the manufacturing of protein therapeutics in mammalian cells to improve productivity, viability, or quality attributes. the direction and extent to which a temperature shift affects productivity and quality may vary depending on the expression host and characteristics of the expressed protein. we demonstrated here that two chinese hamster ovary (cho) clones expressing different human monoclonal antibodies responded diffe ... | 2020 | 31930722 |
| attenuating apoptosis in chinese hamster ovary cells for improved biopharmaceutical production. | chinese hamster ovary (cho) cells are the predominant host cell line for the production of biopharmaceuticals, a growing industry currently worth more than $188 billion usd in global sales. cho cells undergo programmed cell death (apoptosis) following different stresses encountered in cell culture, such as substrate limitation, accumulation of toxic by-products, and mechanical shear, hindering production. genetic engineering strategies to reduce apoptosis in cho cells have been investigated with ... | 2020 | 31930480 |
| kinetic modeling of chinese hamster ovary cell culture: factors and principles. | as a host for therapeutic protein expression, chinese hamster ovary (cho) cells are widely utilized in the mainstream biopharmaceutical industry. cell culture process development plays an important role in transitioning laboratory research to manufacturing. among different mathematic tools, kinetic modeling is commonly achieved through analyzing cell culture data to design process parameters, optimize media, and scale up bioreactors. in this review, we examine key factors for upstream process de ... | 2020 | 31928250 |
| targeted optical imaging of the glucagonlike peptide 1 receptor using exendin-4-irdye 800cw. | the treatment of choice for insulinomas and focal lesions in congenital hyperinsulinism (chi) is surgery. however, intraoperative detection can be challenging. this challenge could be overcome with intraoperative fluorescence imaging, which provides real-time lesion detection with a high spatial resolution. here, a novel method for targeted near-infrared (nir) fluorescence imaging of glucagonlike peptide 1 receptor (glp-1r)-positive lesions, using the glp-1 agonist exendin-4 labeled with irdye 8 ... | 2020 | 31924726 |
| different localization of lysosomal-associated membrane protein 1 (lamp1) in mammalian cultured cell lines. | lysosomal-associated membrane protein 1 (lamp1) mainly localizes to lysosomes and late endosomes. we herein investigated the intracellular localization of lysosomal membrane proteins in five mammalian cultured cell lines. rat lamp1 fused to enhanced green fluorescent protein (egfp) mostly accumulated at a particular cytoplasmic area and barely co-localized with lysotracker® red dnd-99 in golden hamster kidney bhk-21 cells and chinese hamster ovary cho-k1 cells. golden hamster, chinese hamster, a ... | 2020 | 31907597 |
| choice of host cell line is essential for the functional glycosylation of the fc region of human igg1 inhibitors of influenza b viruses. | abs are glycoproteins that carry a conserved n-linked carbohydrate attached to the fc whose presence and fine structure profoundly impacts on their in vivo immunogenicity, pharmacokinetics, and functional attributes. the host cell line used to produce igg plays a major role in this glycosylation, as different systems express different glycosylation enzymes and transporters that contribute to the specificity and heterogeneity of the final igg-fc glycosylation profile. in this study, we compare tw ... | 2020 | 31907284 |
| direct optical detection of cell density and viability of mammalian cells by means of uv/vis spectroscopy. | the critical process parameters cell density and viability during mammalian cell cultivation are assessed by uv/vis spectroscopy in combination with multivariate data analytical methods. this direct optical detection technique uses a commercial optical probe to acquire spectra in a label-free way without signal enhancement. for the cultivation, an inverse cultivation protocol is applied, which simulates the exponential growth phase by exponentially replacing cells and metabolites of a growing ch ... | 2020 | 31897554 |
| genome-scale reconstructions of the mammalian secretory pathway predict metabolic costs and limitations of protein secretion. | in mammalian cells, >25% of synthesized proteins are exported through the secretory pathway. the pathway complexity, however, obfuscates its impact on the secretion of different proteins. unraveling its impact on diverse proteins is particularly important for biopharmaceutical production. here we delineate the core secretory pathway functions and integrate them with genome-scale metabolic reconstructions of human, mouse, and chinese hamster ovary cells. the resulting reconstructions enable the c ... | 2020 | 31896772 |
| summation of disinfection by-product cho cell relative toxicity indices: sampling bias, uncertainty, and a path forward. | the cyto- and genotoxic potencies of disinfection by-products (dbps) have been evaluated in published literature by measuring the response of exposed chinese hamster ovary cells. in recent publications, dbp concentrations divided by their individual toxicity indices are summed to predict the relative toxicity of a water sample. we hypothesized that the omission or inclusion of certain dbps over others is equivalent to statistical sampling bias and may result in biased conclusions. to test this h ... | 2020 | 31894210 |
| robust analytical methods for the accurate quantification of the total biomass composition of mammalian cells. | biomass composition is an important input for genome-scale metabolic models and has a big impact on their predictive capabilities. however, researchers often rely on generic data for biomass composition, e.g. collected from similar organisms. this leads to inaccurate predictions, because biomass composition varies between different cell lines, conditions, and growth phases. in this chapter we present protocols for the determination of the biomass composition of chinese hamster ovary (cho) cells. ... | 2020 | 31893373 |
| small-scale bioreactor supports high density hek293 cell perfusion culture for the production of recombinant erythropoietin. | process intensification in mammalian cell culture-based recombinant protein production has been achieved by high cell density perfusion exceeding 108 cells/ml in the recent years. as the majority of therapeutic proteins are produced in chinese hamster ovary (cho) cells, intensified perfusion processes have been mainly developed for this type of host cell line. however, the use of cho cells can result in non-human posttranslational modifications of the protein of interest, which may be disadvanta ... | 2020 | 31891733 |
| cytotoxicity and genotoxicity of dmabee, a co-photoinitiator of resin polymerization, on cho-k1 cells: role of redox and carboxylesterase. | the 4-dimethylaminobenzoic acid ethyl ester (dmabee) is an important co-initiator for resin polymerization in dental resinous materials. as a radical forming chemical with high lipophilicity, the genotoxicity and cytotoxicity of dmabee deserve prudent investigation. in this study, we found that dmabee reduced the viability and proliferation of chinese hamster ovary (cho-k1) cells in a dose-dependent manner, and altered cell morphology at higher concentrations. g0/g1 cell cycle arrest was induced ... | 2020 | 31880385 |
| unstructured regions in ire1α specify bip-mediated destabilisation of the luminal domain dimer and repression of the upr. | coupling of endoplasmic reticulum (er) stress to dimerisation-dependent activation of the upr transducer ire1 is incompletely understood. whilst the luminal co-chaperone erdj4 promotes a complex between the hsp70 bip and ire1's stress-sensing luminal domain (ire1ld) that favours the latter's monomeric inactive state and loss of erdj4 de-represses ire1, evidence linking these cellular and in vitro observations is presently lacking. we report that enforced loading of endogenous bip onto endogenous ... | 2019 | 31873072 |
| hif-1 signaling pathway implicated in phenotypic instability in a chinese hamster ovary production cell line. | monitoring genotypic and phenotypic stability is crucial during the development of recombinant chinese hamster ovary (cho) cell lines. although genotypic instability is well-studied, there are few reports on phenotypic instability. here, a case study of two clonal cell lines derived from pfizer's site-specific integration expression platform that expresses the same monoclonal antibody is described. it is shown that both cell lines (herein referred to as "cell line a" and "cell line b") are genot ... | 2020 | 31872551 |
| contactless electroporation induced by high intensity pulsed electromagnetic fields via distributed nanoelectrodes. | pulsed electric fields (pefs) can be used to transiently increase cell membrane permeability in procedures ranging from gene therapy to tumor eradication. although very efficient, pef-based therapies generally require the use of invasive electrodes, which cause pain and tissue damage. an emerging noninvasive, contactless alternative to pefs are high intensity pulsed electromagnetic fields (hi-pemf), whereby the electric field inside the tissue is induced remotely by external pulsed magnetic fiel ... | 2020 | 31869699 |
| acquired glanzmann thrombasthenia associated with platelet desialylation. | the notable discrepancy between platelet count and bleeding manifestations in immune thrombocytopenia (itp) patients with acquired glanzmann thrombasthenia (gt) has been described. | 2020 | 31869497 |
| design and synthesis of novel ferrocene-quinoline conjugates and evaluation of their electrochemical and antiplasmodium properties. | the tropical disease malaria is responsible for more than 400,000 deaths annually, especially in southeast asia and africa. although the number of malaria cases is declining, there still is an urgent need for novel antimalarial agents. the emergence of hybrid antimalarial agents and the precedence set by the antimalarial drug ferroquine (fq) prompted us to design new ferrocene-containing quinoline structures. herein, we report the efficient synthesis of three different series of ferrocene-quinol ... | 2020 | 31865015 |
| investigating toxicity of urban road deposited sediments using chinese hamster ovary cells and chlorella pyrenoidosa. | road deposited sediments (rds) is the key carrier of pollutants in the urban road stormwater processes and hence has been seen as an important pollutant source of urban road stormwater. although many research studies have focused on rds and pollutants attached to rds, the investigation on rds toxicity is very limited. toxicity test can permit an overall assessment on whether the rds polluted stormwater can be safely reused. this paper used two living organisms, namely chinese hamster ovary (cho) ... | 2020 | 31864047 |
| how to produce mabs in a cube-shaped stirred single-use bioreactor at 200 l scale. | single-use bioreactors have increasingly been used in recent years, for both research and development as well as industrial production, especially in mammalian cell-based processes. among the numerous single-use bioreactors available today, wave-mixed bags and stirred systems dominate. wave-mixed single-use bioreactors are the system of choice for inoculum production, while stirred single-use bioreactors are most often preferred for antibody expression. for this reason, the present chapter descr ... | 2020 | 31858468 |
| orbitally shaken single-use bioreactor for animal cell cultivation: fed-batch and perfusion mode. | increasing the cultivation volume from small to large scale can be a rather complex and challenging process when the method of aeration and mixing is different between scales. orbitally shaken bioreactors (osbs) utilize the same hydrodynamic principles that define the success of smaller-scale cultures, which are developed on an orbitally shaken platform, and can simplify scale-up. here we describe the basic working principles of scale-up in terms of the volumetric oxygen transfer coefficient (kl ... | 2020 | 31858465 |
| using a parallel micro-cultivation system (micro-matrix) as a process development tool for cell culture applications. | micro-bioreactors appear frequently in today's biotechnology industry as screening and process development tools for cell culture applications. the micro-bioreactor's small volume allows for a high throughput, and when compared to other small-scale systems, such as microtiter plates, its measurement and control capabilities offer a much better insight into the bioprocess. applikon's micro-matrix is one of the micro-bioreactors that are commercially available today. the micro-matrix system consis ... | 2020 | 31858463 |
| high throughput, efficacious gene editing & genome surveillance in chinese hamster ovary cells. | chinese hamster ovary (cho) cells are a common tool utilized in bioproduction and directed genome engineering of cho cells is of great interest to enhance recombinant cell lines. until recently, this focus has been challenged by a lack of efficacious, high throughput, and low-cost gene editing modalities and screening methods. in this work, we demonstrate an improved method for gene editing in cho cells using crispr rnps and characterize the endpoints of cas9 and zfn mediated genetic engineering ... | 2019 | 31856197 |
| reinvestigation of disulfide-bonded oligomeric forms of the unfolded protein response transducer atf6. | atf6α is an endoplasmic reticulum (er)-embedded transcription factor which is rapidly activated by er stress, and a major regulator of er chaperone levels in vertebrates. we previously suggested that atf6α occurs as a monomer, dimer and oligomer in the unstressed er of chinese hamster ovary cells due to the presence of two evolutionarily conserved cysteine residues in its luminal region (c467 and c618), and showed that atf6α is reduced upon er stress, such that only reduced monomer atf6α is tran ... | 2020 | 31852864 |
| biallelic loss-of-function mutations in jam2 cause primary familial brain calcification. | primary familial brain calcification is a monogenic disease characterized by bilateral calcifications in the basal ganglia and other brain regions, and commonly presents motor, psychiatric, and cognitive symptoms. currently, four autosomal dominant (slc20a2, pdgfrb, pdgfb, xpr1) and one autosomal recessive (myorg) causative genes have been identified. compared with patients with autosomal dominant primary familial brain calcification, patients with the recessive form of the disease present with ... | 2020 | 31851307 |
| investigation and circumvention of transfection inhibition by ferric ammonium citrate in serum-free media for chinese hamster ovary cells. | while reliable transfection methods are essential for chinese hamster ovary (cho) cell line engineering, reduced transfection efficiencies have been observed in several commercially prepared media. in this study, we aimed to assess common media additives that impede efficiency mediated by three chemical transfection agents: liposomal-based (lipofectamine 2000), polymer-based (transit-x2), and lipopolyplex-based (transit-pro). an in-house gfp-expressing vector and serum-free medium (bcr-f12: deve ... | 2020 | 31850663 |
| insights into virus inactivation by polysorbate 80 in the absence of solvent. | triton x-100 has long been used either alone or in combination with solvent to inactivate enveloped viruses in biopharmaceutical manufacturing. however, european chemicals agency (echa) officially placed triton x-100 on the annex xiv authorization list in 2017 because 4-(1,1,3,3-tetramethylbutyl) phenol, a degradation product of triton x-100, is of harmful endocrine disrupting activities. as a result, any use of triton x-100 in the european economic area would require an echa issued authorizatio ... | 2020 | 31846227 |
| endothelial intercellular cell adhesion molecule 1 contributes to cell aggregate formation in cho cells cultured in serum-free media. | chinese hamster ovary (cho) cells have been adapted to grow in serum-free media and in suspension culture to facilitate manufacturing needs. some cho cell lines, however, tend to form cell aggregates while being cultured in suspension. this can result in reduced viability and capacity for single cell cloning (scc) via limiting dilution, and process steps to mitigate cell aggregate formation, for example, addition of anti-cell-aggregation agents. in this study, we have identified endothelial inte ... | 2020 | 31846224 |
| surrogates for the removal by ozonation of the cytotoxicity and dna double-strand break effects of wastewater on mammalian cells. | effluents from wastewater treatment plants (wwtps) may contain various pollutants with potential toxic effects. ozonation is widely applied to purify wastewater, which may influence the toxicity and water quality indices simultaneously. the main goal of this study was to reveal influence of ozonation on toxicity of wwtp effluents and to find the surrogates for toxicity changes. cytotoxicity and dna double-strand break (dsb) effect of wwtp effluents were measured based on chinese hamster ovary (c ... | 2020 | 31841803 |
| targeting ovarian cancer cell cytotoxic drug resistance phenotype with xanthium strumarium l. extract. | emerging drugs aim at targeting the genomic integrity and replication machinery in ovarian cancer. while the antiproliferative activity of xanthium strumarium l. extract (xfc), a traditional herbal medicine, is believed to alter the mitotic apparatus of chinese hamster ovary epithelial cells, its capacity to target and overcome the chemoresistance phenotype in ovarian cancer is unknown. among the cancer cell lines tested, we found that the best proliferation inhibitory effect for xfc was against ... | 2019 | 31827554 |
| role of a new bioassay for thyroid-stimulating antibodies (aequorin tsab) in graves' ophthalmopathy. | graves' ophthalmopathy (go) is characterized by an autoimmune reaction against thyrotropin (tsh) receptors and is diagnosed by tsh receptor antibody (trab). a novel assay for thyroid-stimulating antibody (tsab) was recently introduced using a frozen chinese hamster ovary cell line expressing tsh receptors, cyclic adenosine monophosphate (camp)-gated calcium channel, and aequorin (aequorin tsab). the aim of this study was to evaluate the role of aequorin tsab in go. we studied 136 japanese patien ... | 2020 | 31827052 |
| structure-effect relationships of novel semi-synthetic cannabinoid derivatives. | background: as a library of cannabinoid (cb) derivatives with (-)-trans-cannabidiol (cbd) or (-)-trans-cannabidivarin (cbdv) scaffold, we synthesized nine novel cannabinoids: 2-hydroxyethyl cannabidiolate (2-hec), 2-hydroxypentyl cannabidiolate (2-hpc), 2,3-dihydroxypropyl cannabidiolate (gcbd), cyclohexyl cannabidiolate (chc), n-hexyl-cannabidiolate (hc), 2-(methylsulfonamido)ethyl cannabidiolate (nmsc), 2-hydroxyethyl cannabidivarinolate (2-hecbdv), cyclohexyl cannabidivarinolate (chcbdv), and ... | 2019 | 31824305 |
| effects of cysteine, asparagine, or glutamine limitations in chinese hamster ovary cell batch and fed-batch cultures. | amino acid availability is a key factor that can be controlled to optimize the productivity of fed-batch cultures. to study amino acid limitation effects, a serum-free chemically defined basal medium was formulated to exclude the amino acids that became depleted in batch culture. the effect of limiting glutamine, asparagine, and cysteine on the cell growth, metabolism, antibody productivity, and product glycosylation was investigated in three chinese hamster ovary (cho) cell lines (cho-dxb11, ch ... | 2020 | 31823468 |
| polymorphisms and endometriosis: a systematic review and meta-analyses. | endometriosis is an estrogen-dependent gynecological disorder that affects at least 10% of women of reproductive age. it may lead to infertility and non-specific symptoms such as chronic pelvic pain. endometriosis screening and diagnosis are difficult and time-consuming. late diagnosis (with a delay ranging from 3.3 to 10.7 years) is a major problem and may contribute to disease progression and a worse response to treatment once initiated. efficient screening tests might reduce this diagnostic d ... | 2020 | 31821471 |
| oxidative stress-alleviating strategies to improve recombinant protein production in cho cells. | large scale biopharmaceutical production of biologics relies on the overexpression of foreign proteins by cells cultivated in stirred tank bioreactors. it is well recognized and documented fact that protein overexpression may impact host cell metabolism and that factors associated with large scale culture, such as the hydrodynamic forces and inhomogeneities within the bioreactors, may promote cellular stress. the metabolic adaptations required to support the high-level expression of recombinant ... | 2020 | 31814104 |
| development of rolled scaffold for high-density adherent cell culture. | bioreactors for large-scale culture of mammalian cells are playing vital roles in biotechnology and bioengineering. various bioreactors have been developed, but their capacity and efficiency are often limited by insufficient mass transfer rate and high shear stress. a rolled scaffold (rs) is a fully defined scaffold for high-density adherent culture of mammalian cells. the rs is a polymer film with spacers, that is rolled into a cylinder with a pre-determined gap between each turn. cells are cul ... | 2019 | 31813056 |
| radioprotective effect of hydroxyl radical scavengers on prokaryotic and eukaryotic cells under various gamma irradiation conditions. | the influence of various hydroxyl radical scavengers such as methanol, ethanol and dimethyl sulfoxide on radiation sensitivity of prokaryotic cells (bacteria escherichia coli) and eukaryotic cells (yeast saccharomyces cerevisiae and v79 cells-chinese hamster pulmonary fibroblasts) irradiated by 60co gamma radiation was investigated. the dependence of radiation sensitivity on dose rate in range from 1.8 to 100 gy h-1 was evaluated. survival of cells irradiated by increasing dose rates was followe ... | 2019 | 31812995 |
| microrna-504 functions as a tumor suppressor in oral squamous cell carcinoma through inhibiting cell proliferation, migration and invasion by targeting cdk6. | microrna (mirna), as tumor suppressor or oncogene, is involved in the regulation of tumor development. however, the role of mirna in human oral squamous cell carcinoma (oscc) is less well understood. in our previous studies, we have successfully established a chinese hamster oral squamous cell carcinoma animal model and constructed a mirna differential expression profile, and screened out the abnormal expressed gene (mir-504). the purpose of this study was to investigate the regulatory mechanism ... | 2020 | 31812760 |
| engineering of chinese hamster ovary cell lipid metabolism results in an expanded er and enhanced recombinant biotherapeutic protein production. | chinese hamster ovary (cho) cell expression systems have been exquisitely developed for the production of recombinant biotherapeutics (e.g. standard monoclonal antibodies, mabs) and are able to generate efficacious, multi-domain proteins with human-like post translational modifications at high concentration with appropriate product quality attributes. however, there remains a need for development of new cho cell expression systems able to produce more challenging secretory recombinant biotherape ... | 2020 | 31805379 |
| dr10601, a novel recombinant long-acting dual glucagon-like peptide-1 and glucagon receptor agonist for the treatment of obesity and type 2 diabetes mellitus. | both glucagon-like peptide-1 (glp-1) and glucagon (gcg) belong to the incretin family. this study aimed to investigate the pharmacokinetics and pharmacodynamics of dr10601, a fully recombinant hybrid peptide with dual glp-1/gcg receptor agonistic activity. | 2020 | 31786794 |
| modelling of hollow fiber membrane bioreactor for mammalian cell cultivation using computational hydrodynamics. | the hollow fiber membrane bioreactor (hfmb) has been investigated for the cultivation of mammalian chinese hamster ovary cell expansion. the experiments were carried out in petri's dishes and in the hollow fiber membrane bioreactor having 20 fibers (s2025 from fibercell systems). the approach to hfmb modelling which combines the model of cell growth kinetics and hydrodynamics has been proposed. the hydrodynamic model is made using ansys fluent software. the mathematical model of hfmb was develop ... | 2020 | 31786664 |
| knockout of sialidase and pro-apoptotic genes in chinese hamster ovary cells enables the production of recombinant human erythropoietin in fed-batch cultures. | sialic acid, a terminal monosaccharide present in n-glycans, plays an important role in determining both the in vivo half-life and the therapeutic efficacy of recombinant glycoproteins. low sialylation levels of recombinant human erythropoietin (rhepo) in recombinant chinese hamster ovary (rcho) cell cultures are considered a major obstacle to the production of rhepo in fed-batch mode. this is mainly due to the accumulation of extracellular sialidases released from the cells. to overcome this hu ... | 2020 | 31785386 |
| bioinformatic identification of chinese hamster ovary (cho) cold-shock genes and biological evidence of their cold-inducible promoters. | lower cultivation temperature dramatically affects cell growth and cellular productivity in chinese hamster ovary cells (cho) and is often used in industrial applications with the aim to enhance productivity. cold-inducible proteins whose activity is induced at lower temperature play an important role in understanding the mechanisms of cold-induced changes in gene expression. one of these mechanisms is increased transcription of specific target genes controlled by sequence elements in cold-induc ... | 2020 | 31785035 |
| novel kcnj10 compound heterozygous mutations causing east/sesame-like syndrome compromise potassium channel function. | inwardly rectifying k+ channel 4.1 (kir4.1), encoded by kcnj10, is a member of the inwardly rectifying potassium channel family. in the brain, kir4.1 is predominant in astrocytic glia and accounts for the spatial buffering of k+ released by neurons during action potential propagation. a number of studies have shown that mutations in kcnj10 are associated with sesame/east syndrome, which is characterized by seizures, ataxia, sensorineural deafness, and electrolyte imbalance. herein, we identified ... | 2019 | 31781151 |
| evidence for diversity in the activation of the melanocortin 2 receptor: a study on gar, elephant shark and stingray mc2rs. | the melanocortin-2 receptor (mc2r) is a critical component of the hpi and hpa axes of cartilaginous fishes, teleosts and tetrapods. studies on teleost and tetrapod orthologs suggest two contact sites between acth and the receptor involving the following motifs on acth: h6f7r8w9 and k15k16r17r18p19. using spotted gar (g) mc2r as a representative bony fish mc2r ortholog, we found that activation of gmc2r in chinese hamster ovary (cho) cells was diminished following stimulation of the transfected c ... | 2020 | 31778725 |
| engineering of an iranian bordetella pertussis strain producing inactive pertussis toxin. | introduction. differences between the genomic and virulence profile of bordetella pertussis circulating strains and vaccine strains are considered as one of the important reasons for the resurgence of whooping cough (pertussis) in the world. genetically inactivated b. pertussis is one of the new strategies to generate live-attenuated vaccines against whooping cough.aim. the aim of this study was to construct a b. pertussis strain based on a predominant profile of circulating iranian isolates tha ... | 2020 | 31778110 |
| distance effect characteristic of the matrix attachment region increases recombinant protein expression in chinese hamster ovary cells. | previously, we have found that the matrix attachment region (mar) may confer a 'distance effect' on transgene expression. this work aims to systematically explore the increased transgene expression in transfected chinese hamster ovary (cho) cells due to the characteristics of mar and its mechanism. | 2020 | 31776751 |
| heparin chromatography as an in vitro predictor for antibody clearance rate through pinocytosis. | the pharmacokinetic (pk) properties of therapeutic antibodies directly affect efficacy, dose and dose intervals, application route and tissue penetration. in indications where health-care providers and patients can choose between several efficacious and safe therapeutic options, convenience (determined by dosing interval or route of application), which is mainly driven by pk properties, can affect drug selection. therapeutic antibodies can have greatly different pk even if they have identical fc ... | 2020 | 31769731 |
| a novel method based on nonparametric regression with a gaussian kernel algorithm identifies the critical components in cho media and feed optimization. | as the composition of animal cell culture medium becomes more complex, the identification of key variables is important for simplifying and guiding the subsequent medium optimization. however, the traditional experimental design methods are impractical and limited in their ability to explore such large feature spaces. therefore, in this work, we developed a nrgk (nonparametric regression with gaussian kernel) method, which aimed to identify the critical components that affect product titres duri ... | 2020 | 31754859 |
| unaccompanied mechanosensory domain mediates low expression of glycoprotein ibα: implications for bernard-soulier syndrome. | disruption of protein folding or inter-subunit interactions in the platelet glycoprotein (gp)ib-ix complex leads to its abnormally low expression in the plasma membrane, the hallmark of bernard-soulier syndrome (bss). | 2020 | 31749281 |
| ercc1 c8092a polymorphism predicts fair survival outcome in japanese patients with pharyngo-laryngeal squamous cell carcinoma. | to evaluate the prognostic significance of dna excision repair gene polymorphisms, excision repair cross-complementation group 1 (ercc1) and x-ray repair complementing defective repair in chinese hamster cells 1 (xrcc1) polymorphisms were investigated in japanese patients with head and neck squamous cell carcinoma (hnscc). | 2020 | 31749055 |
| developing an equivalent toxicity area approach to comparing toxicity of urban road deposited sediments. | urban road stormwater has been seen as an alternative water resource for mitigating water scarcity. road deposited sediments (rds) is the primary source of road stormwater pollution since many toxic pollutants are attached to rds and eventually enter stormwater runoff when rainfall events occur. this could threaten road stormwater reuse safety. this study developed an innovative equivalent toxicity area (eta) approach to comparing overall toxicity of rds among different road sites. in the eta ap ... | 2020 | 31744684 |
| assuring clonality on the beacon digital cell line development platform. | during biomanufacturing cell lines development, the generation and screening for single-cell derived subclones using methods that enable assurance of clonal derivation can be resource- and time-intensive. high-throughput miniaturization, automation, and analytic strategies are often employed to reduce such bottlenecks. the beacon platform from berkeley lights offers a strategy to eliminate these limitations through culturing, manipulating, and characterizing cells on custom nanofluidic chips via ... | 2020 | 31743597 |
| resveratrol addition to chinese hamster ovary cell culture media: the effect on cell growth, monoclonal antibody synthesis, and its chemical modification. | the effect of the addition of resveratrol to cell culture media during the production of monoclonal antibodies was investigated. treatments of chinese hamster ovary (cho) cells expressing immunoglobulin g (igg) with 25 and 50 μm resveratrol showed that resveratrol was capable of slowing cell growth while almost doubling cell-specific productivity to 4.7 ± 0.6 pg igg/cell·day, resulting in up to a 1.37-fold increase of the final igg titer. a resveratrol concentration of 50 μm slowed the progressi ... | 2020 | 31742929 |
| comparative cytotoxicity of halogenated aromatic dbps and implications of the corresponding developed qsar model to toxicity mechanisms of those dbps: binding interactions between aromatic dbps and catalase play an important role. | halogenated aromatic disinfection byproducts (dbps) are a new group of emerging dbps identified recently. they have been detected in disinfected drinking water, wastewater effluents, recreational water and oil/gas produced water, at concentrations of ng/l to μg/l in general. previously studies have demonstrated that most of them can induce developmental toxicity and growth inhibition in aquatic organisms based on in vivo bioassays. in this study, to further understand the adverse effects of arom ... | 2020 | 31739241 |
| human cyp1b1-dependent genotoxicity of dioxin-like polychlorinated biphenyls in mammalian cells. | polychlorinated biphenyls (pcbs) are persistent organic pollutants and human carcinogens. it was reported that rat cyp1a1 and catfish cyp1a can hydroxylate 3,3',4,4',5-pentachlorobiphenyl (pcb 126) and 3,3',4,4'-tetrachlorobiphenyl (pcb 77), while potential roles of other cyp1 enzymes in the metabolism of dioxin-like (dl) pcbs remain unconfirmed. in this study, three representative dl-pcbs, i.e., pcb 77, pcb 126, and 3,4,4',5-tetrachlorobiphenyl (pcb 81), were investigated on their genotoxicity ... | 2020 | 31733235 |
| a human expression system based on hek293 for the stable production of recombinant erythropoietin. | mammalian host cell lines are the preferred expression systems for the manufacture of complex therapeutics and recombinant proteins. however, the most utilized mammalian host systems, namely chinese hamster ovary (cho), sp2/0 and ns0 mouse myeloma cells, can produce glycoproteins with non-human glycans that may potentially illicit immunogenic responses. hence, we developed a fully human expression system based on hek293 cells for the stable and high titer production of recombinant proteins by fi ... | 2019 | 31727983 |
| an agonist radioligand for the proinflammatory lipid-activated g protein-coupled receptor gpr84 providing structural insights. | the orphan g protein-coupled receptor (gpcr) gpr84 is expressed on immune cells mediating proinflammatory and immunostimulatory effects. in this study, we prepared the fully efficacious, nonbiased gpr84 agonist 6-hexylamino-2,4(1h,3h)-pyrimidinedione (6) in tritium-labeled form ([3h]psb-1584) by hydrogenation of a hexenyl-substituted precursor with tritium gas. the radioligand was characterized by kinetic, saturation, and competition assays using membranes of chinese hamster ovary cells recombin ... | 2020 | 31721581 |
| an easy and fast method for production of chinese hamster ovary cell line expressing and secreting human recombinant activin a. | growth factors are key elements of embryonic stem cell (esc) research. cell line development in eukaryotes is a time-consuming procedure which usually takes 12-18 months. here, we report an easy and fast method with which production of chinese hamster ovary (cho) cells that express and secrete recombinant activin a, as a major growth factor in endo/mesoderm differentiation of embryonic stem cells is achieved within 3-4 weeks. | 2020 | 31721527 |
| application of a genome-scale model in tandem with enzyme assays for identification of metabolic signatures of high and low cho cell producers. | biopharmaceutical industrial processes are based on high yielding stable recombinant chinese hamster ovary (cho) cells that express monoclonal antibodies. however, the process and feeding regimes need to be adapted for each new cell line, as they all have a slightly different metabolism and product performance. a main limitation for accelerating process development is that the metabolic pathways underlying this physiological variability are not yet fully understood. this study describes the evol ... | 2019 | 31720213 |
| aqueous extract of forsythia viridissima fruits: acute oral toxicity and genotoxicity studies. | forsythia viridissima fruit, one of forsythiae fructus (ff) is widely used in traditional medicine to treat diverse diseases-related clinical symptoms, including fever, pain, vomiting, nausea, and abscess. however, the safety of ff has not been fully assessed. | 2020 | 31715286 |
| chlorinated effluent organic matter causes higher toxicity than chlorinated natural organic matter by inducing more intracellular reactive oxygen species. | during unplanned indirect potable reuse, treated wastewater that contains effluent organic matter (eom) enters the drinking water source, resulting in different toxicity from natural organic matter (nom) in surface water during chlorination. this study found that, during chlorination, eom formed more total organic halogen (tox) and highly toxic nitrogenous disinfection byproducts (dbps) like dichloroacetonitrile and trichloronitromethane than nom did. oxidative stress including both reactive oxy ... | 2020 | 31710900 |
| cardiotoxic effects of [3-[2-(diethylamino)ethyl]-1h-indol-4-yl] acetate and 3-[2-[ethyl(methyl)amino]ethyl]-1h-indol-4-ol. | two synthetic tryptamines, namely [3-[2-(diethylamino)ethyl]-1h-indol-4-yl] acetate (4-aco-det) and 3-[2-[ethyl(methyl)amino]ethyl]-1h-indol-4-ol (4-ho-met), are abused by individuals seeking recreational hallucinogens. these new psychoactive substances (npss) can cause serious health problems because their adverse effects are mostly unknown. in the present study, we evaluated the cardiotoxicity of 4-aco-det and 4-ho-met using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (mtt ... | 2020 | 31706004 |
| early integration of design of experiment (doe) and multivariate statistics identifies feeding regimens suitable for cho cell line development and screening. | in chinese hamster ovary (cho) cell lines, the establishment of the ideal fed-batch regimen promotes metabolic conditions advantageous for the bioproduction of therapeutic molecules. a tailored, cell line-specific feeding scheme is typically defined during process development (pd) activities, through the incorporation of design of experiment (doe) and late stage cell culture approaches. the feeding during early stage cell line development (cld) was a simplified "one-fits-all" design, inherited f ... | 2019 | 31705334 |
| descubrimiento de la proteína reguladora de apoptosis 2 como determinante en la muerte de células tumorales. | cancer is a multifactorial disease that constitutes a serious public health problem worldwide. prostate cancer advanced stages are associated with the development of androgen-independent tumors and an apoptosis-resistant phenotype that progresses to metastasis. by studying androgen-independent lymphoid nodule carcinoma of the prostate (lncap) cells induced to apoptosis by serum elimination, we identified the activation of a non-selective cationic channel of 23ps conductance that promotes incomin ... | 2019 | 31695224 |
| a high yielding ifnar1 ecd mammalian expression process for use in autoimmune disease drug development. | interferon-alpha receptor 1 (ifnar1) is a target of interest for recombinant biotherapeutics that block the jak/stat pathway. this pathway is believed to play a role in many diseases including hepatitis b and c, herpes simplex, multiple sclerosis, and other autoimmune disorders. by using ifnar1 as a target to block type i ifn from binding to the jak/stat pathway and prevent activation of this target, autoimmune disease progression can be modulated. current ifnar1 extracellular domain (ecd) expre ... | 2020 | 31689498 |
| computer-aided strategies for determining the amino acid composition of medium for chinese hamster ovary cell-based biomanufacturing platforms. | chinese hamster ovary (cho) cells are used for the production of the majority of biopharmaceutical drugs, and thus have remained the standard industry host for the past three decades. the amino acid composition of the medium plays a key role in commercial scale biologics manufacturing, as amino acids constitute the building blocks of both endogenous and heterologous proteins, are involved in metabolic and non-metabolic pathways, and can act as main sources of nitrogen and carbon under certain co ... | 2019 | 31684012 |