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toxicity assessment of gryllus bimaculatus (a type of cricket) glycosaminoglycan.we performed general toxicity studies of gryllus bimaculatus (two-spotted cricket) glycosaminoglycan (gbg), including a single, 4-week repeated oral dose toxicity test in icr mice, and short-term genotoxicity tests. the mutagenic potential of the purified gbg was non-genotoxic when it was evaluated using short-term genotoxicity tests, namely ames, chromosome aberration (ca), and micronuclei (mn) tests. in salmonella typhimurium and escherichia coli assays, gbg did not produce any mutagenic respo ...202033005591
pharmacokinetic prediction of an antibody in mice based on an in vitro cell-based approach using target receptor-expressing cells.in vivo pharmacokinetics (pk) studies using mice and monkeys are the main approaches for evaluating and predicting the pk of antibodies, and there is a strong demand for methods that do not require animal experiments. in this work, we focused on quantitatively predicting the nonlinear pk of an antibody based on cell-based assays. an anti-mouse fc gamma receptor iib antibody was used as a model antibody. to determine the pk parameters related to nonspecific elimination in vivo, the plasma concent ...202033004886
raman spectra-based deep learning: a tool to identify microbial contamination.deep learning has the potential to enhance the output of in-line, on-line, and at-line instrumentation used for process analytical technology in the pharmaceutical industry. here, we used raman spectroscopy-based deep learning strategies to develop a tool for detecting microbial contamination. we built a raman dataset for microorganisms that are common contaminants in the pharmaceutical industry for chinese hamster ovary (cho) cells, which are often used in the production of biologics. using a c ...202033063423
biological activities of a recombinant fortilin from fenneropenaeus merguiensis.human fortilin, an antiapoptotic protein, has also been implicated in several diseases; however, several potential uses of fortilin have also been proposed. bearing the implications of fortilin in mind, fortilin analog, which has no complication with diseases, is required. since a recombinant full-length fortilin from fenneropenaeus merguiensis (rfm-fortilin (fl)) reported only 44% (3e-27) homologous to human fortilin, therefore the biological activities of the fm-fortilin (fl) and its fragments ...202033002062
a defucosylated anti‑cd44 monoclonal antibody 5‑mg2a‑f exerts antitumor effects in mouse xenograft models of oral squamous cell carcinoma.cd44 is widely expressed on the surface of most tissues and all hematopoietic cells, and regulates many genes associated with cell adhesion, migration, proliferation, differentiation, and survival. cd44 has also been studied as a therapeutic target in several cancers. previously, an anti‑cd44 monoclonal antibody (mab), c44mab‑5 (igg1, kappa) was established by immunizing mice with cd44‑overexpressing chinese hamster ovary (cho)-k1 cells. c44mab‑5 recognized all cd44 isoforms, and showed high sen ...202033000243
the quantitative analysis of the mechanism involved in pertussis toxin-mediated cell clustering and its implications in the in vitro quality control of diphtheria tetanus and whole cell pertussis vaccines.some of the adverse side-effects such as leukocytosis, hyperinsulinemia, hypoglycemia and sensitization to histamine, caused by diphtheria, tetanus and whole cell pertussis (dtwp) vaccines are related to the presence of non-inactivated pertussis toxin residues (niptxr). the chinese hamster ovary (cho) cell clustering assay is an in vitro assay to measure niptxr in dtwp vaccines based on the ability of active pertussis toxin (ptx) to cause cellular clustering. to study the biochemical mechanism i ...202033059000
polymer fraction including exosomes derived from chinese hamster ovary cells promoted their growth during serum-free repeated batch culture.while continuous (perfusion) culture of mammalian cells might reduce the reactor size owing to the high cell density, there is the problem of higher medium cost; however, this problem is expected to be solved by the reuse of growth-promoting components in the culture supernatant. the polymer fraction (pf, 10 kda-220 nm) collected from the supernatant of serum-free repeated-batch culture of chinese hamster ovary (cho) cells in not only adhesion but also suspension promoted the cell growth in resp ...202033051156
hypericin, a medicinal compound from st. john's wort, inhibits genotoxicity induced by mutagenic agents in v79 cells.this study evaluated the cytotoxic, genotoxic, and the modulatory effects on dna damage of hypericin in chinese hamster lung fibroblasts (v79 cells). the hypericin is a natural polycyclic quinone, mainly extracted from st. john's wort (hypericum perforatum l.). along with hyperforin, the hypericins are responsible for the antidepressant activity of st. john's wort. cytotoxicity was assessed by the xtt colorimetric assay and the nuclear division index (ndi). the genotoxic activity was studied by ...202033050761
toolbox accelerating glycomics (tag): glycan annotation from maldi-tof ms spectra and mapping expression variation to biosynthetic pathways.glycans present extraordinary structural diversity commensurate with their involvement in numerous fundamental cellular processes including growth, differentiation, and morphogenesis. unlike linear dna and protein sequences, glycans have heterogeneous structures that differ in composition, branching, linkage, and anomericity. these differences pose a challenge to developing useful software for glycomic analysis. to overcome this problem, we developed the novel toolbox accelerating glycomics (tag ...202032998456
synthesis and biological evaluation of antibody drug conjugates based on an antibody expression system: conamax.antibody production for adcs (or in general) is commonly performed by cho-based platforms and limited by volumetric productivity, expensive downstream purification, and extended optimization timelines. the conamax platform is a novel microbial-based protein production and secretion system. a suite of synthetic biology tools have enabled high volumetric productivity (>1 g/l/d) and glycoengineering to produce simple and consistent human-like post-translational modifications. conamax can be enginee ...202032280859
safety evaluation of water extract of gastrodia elata blume: genotoxicity and 28-day oral toxicity studies.water extract of gastrodia elata blume (wge) has great potential as an anti-depressant and could be developed as a functional food. this study aims to assess the safety of wge using in vitro and in vivo genotoxicity assays and a 28-day oral toxicity study. results from a bacterial reverse mutation assay (ames test) using five salmonella typhimurium strains (ta98, ta100, ta102, ta1535, and ta1537) with or without metabolic activation (s9 system) showed that wge did not induce mutagenicity. nor di ...202032278877
spike protein recognition of mammalian ace2 predicts the host range and an optimized ace2 for sars-cov-2 infection.sars-cov-2 causes the recent global covid-19 public health emergency. ace2 is the receptor for both sars-cov-2 and sars-cov. to predict the potential host range of sars-cov-2, we analyzed the key residues of ace2 for recognizing s protein. we found that most of the selected mammals including pets (dog and cat), pangolin and circetidae mammals remained the most of key residues for association with s protein from sars-cov and sars-cov-2. the interaction interface between cat/dog/pangolin/chinese h ...202032201080
branched-chain amino acid catabolism promotes thrombosis risk by enhancing tropomodulin-3 propionylation in platelets.branched-chain amino acids (bcaas), essential nutrients including leucine, isoleucine, and valine, serve as a resource for energy production and the regulator of important nutrient and metabolic signals. recent studies have suggested that dysfunction of bcaa catabolism is associated with the risk of cardiovascular disease. platelets play an important role in cardiovascular disease, but the functions of bcaa catabolism in platelets remain unknown.202032200651
cytotoxicity of halogenated tyrosyl compounds, an emerging class of disinfection byproducts.halogenated amino acids and peptides are an emerging class of disinfection byproducts (dbps), having been detected in drinking water and in washed food products. however, the toxicological significance of these emerging dbps remains unclear. in this study, the cytotoxicity of eight halogenated tyrosyl compounds was investigated in chinese hamster ovary (cho) cells using real-time cell analysis (rtca). dihalogenated tyrosyl compounds are more cytotoxic than their monohalogenated analogues. the cy ...202032200635
cell-free protein synthesis: a promising option for future drug development.proteins are the main source of drug targets and some of them possess therapeutic potential themselves. among them, membrane proteins constitute approximately 50% of the major drug targets. in the drug discovery pipeline, rapid methods for producing different classes of proteins in a simple manner with high quality are important for structural and functional analysis. cell-free systems are emerging as an attractive alternative for the production of proteins due to their flexible nature without a ...202032198631
a new class of quadruplex dna-binding nickel schiff base complexes.we have prepared six new nickel schiff base complexes via reactions of substituted benzophenones with different diamines in the presence of nickel(ii). these new complexes were then reacted with 1-(2-choroethyl)piperidine to afford a further six novel nickel(ii) schiff base complexes bearing pendant ethylpiperidine groups. the complexes bearing the ethylpiperidine moieties had greater solubility in water, and were therefore suitable for use in dna binding experiments. esi mass spectra of solutio ...202032219227
defects in galactose metabolism and glycoconjugate biosynthesis in a udp-glucose pyrophosphorylase-deficient cell line are reversed by adding galactose to the growth medium.udp-glucose (udp-glc) is synthesized by ugp2-encoded udp-glc pyrophosphorylase (ugp) and is required for glycoconjugate biosynthesis and galactose metabolism because it is a uridyl donor for galactose-1-p (gal1p) uridyltransferase. chinese hamster lung fibroblasts harboring a hypomrphic ugp(g116d) variant display reduced udp-glc levels and cannot grow if galactose is the sole carbon source. here, these cells were cultivated with glucose in either the absence or presence of galactose in order to ...202032188137
changes of resurgent na+ currents in the nav1.4 channel resulting from an scn4a mutation contributing to sodium channel myotonia.myotonia congenita (mc) is a rare disorder characterized by stiffness and weakness of the limb and trunk muscles. mutations in the scn4a gene encoding the alpha-subunit of the voltage-gated sodium channel nav1.4 have been reported to be responsible for sodium channel myotonia (scm). the nav1.4 channel is expressed in skeletal muscles, and its related channelopathies affect skeletal muscle excitability, which can manifest as scm, paramyotonia and periodic paralysis. in this study, the missense mu ...202032276507
assessing additivity of cytotoxicity associated with disinfection byproducts in potable reuse and conventional drinking waters.recent studies used the sum of the measured concentrations of individual disinfection byproducts (dbps) weighted by their chinese hamster ovary (cho) cell cytotoxicity lc50 values to estimate the dbp-associated cytotoxicity of disinfected waters. this approach assumed that cytotoxicity was additive rather than synergistic or antagonistic. in this study, we evaluated whether this assumption was valid for mixtures containing dbps at the concentration ratios measured in authentic disinfected waters ...202032275830
the effect of amino acid supplementation in an industrial chinese hamster ovary process.the main goal in biosimilar development is to increase chinese hamster ovary (cho) viability and productivity while maintaining product quality. despite media and feed optimization during process development, depletion of amino acids still occurs. the aim of the work was to optimize an existing industrial fed batch process by preventing shortage of amino acids and to gather knowledge about cho metabolism. several process outputs were evaluated such as cell metabolism, cell viability, monoclonal ...202032274904
temperature down-shift modifies expression of upr-/erad-related genes and enhances production of a chimeric fusion protein in cho cells.low culture temperature enhances the cell-specific productivity of chinese hamster ovary (cho) cells expressing varied recombinant (r-) proteins, but the mechanisms remain unclear. regulation of unfolded protein response (upr) pathway genes, such as transcriptional regulatory factors and endoplasmic reticulum (er)-resident proteins, appear to be involved in the improvements of r-protein production under low temperature conditions. the transcriptional regulation of upr-specific targets is studied ...202032271992
validation and identification of reference genes in chinese hamster ovary cells for fc-fusion protein production.in order to reveal potential genotype-phenotype relationship, rt-qpcr reactions are frequently applied which require validated and reliable reference genes. with the investigation on long-term passage and fed-batch cultivation of cho cells producing an fc-fusion protein, four new reference genes-akr1a1, gpx1, aprt, and rps16, were identified from 20 candidates with the aid of genorm, normfinder, bestkeeper, and δct programs and methods. this article provided more verified options in reference ge ...202032216463
pharmacological properties of whale shark (rhincodon typus) melanocortin-2 receptor and melancortin-5 receptor: interaction with mrap1 and mrap2.in the current study, the whale shark (ws; rhincodon typus) melanocortin-2 receptor (mc2r) co-expressed with wsmrap1 in chinese hamster ovary (cho) cells could be stimulated in a dose dependent manner by acth(1-24) with an ec50 of 2.6 × 10-10 m ± 9.7 × 10-11. when the receptor was expressed alone, stimulation was only observed at [10-6 m]. a comparable increase in sensitivity to stimulation by srdes-ac-αmsh was also observed when the receptor was co-expressed with wsmrap1. in addition, co-expres ...202032213301
screening of new cell cycle suppressive compounds from marine-derived microorganisms in chinese hamster ovary cells.monoclonal antibodies (mabs) are active pharmaceutical ingredients in antibody drugs, produced mainly using recombinant chinese hamster ovary (cho) cells. the regulation of recombinant cho cell proliferation can improve the productivity of heterologous proteins. chemical compound approaches for cell cycle regulation have the advantages of simplicity and ease of use in industrial processes. however, cho cells have genetic and phenotypic diversity, and the effects of such compounds might depend on ...202032253091
development of magoh protein-overexpressing hek cells for optimized therapeutic protein production.in the pharmaceutical industry, the need for high levels of protein expression in mammalian cells has prompted the search for new strategies, including technologies to obtain cells with improved mechanisms that enhance its transcriptional activity, folding, or protein secretion. chinese hamster ovary (cho) cells are by far the most used host cell for therapeutic protein expression. however, these cells produce specific glycans that are not present in human cells and therefore potentially immunog ...202032249976
circulating ficolin-2 and ficolin-3 form heterocomplexes.the complement system constitutes an important part of the innate immune system. the collectins and the ficolins are soluble pattern recognition molecules that contribute to complement activation via the lectin pathway. during previous experiments with ficolin-2 and ficolin-3, we have observed that the molecules may interact. we therefore hypothesized the existence of stable ficolin-2/-3 heterocomplexes. we could demonstrate ficolin-2/-3 heterocomplexes in normal human serum and plasma by elisa ...202032094208
ascorbic acid 2-glucoside pretreatment protects cells from ionizing radiation, uvc, and short wavelength of uvb.ascorbic acid 2-glucoside (aa2g), glucosylated ascorbic acid (aa), has superior properties for bioavailability and stability compared to aa. although aa2g has shown radioprotective properties similar to aa, effects for uv light, especially uvc and uvb, are not studied. aa2g was tested for cytotoxicity and protective effects against ionizing radiation, uvc, and broadband and narrowband uvb in chinese hamster ovary (cho) cells and compared to aa and dimethyl sulfoxide (dmso). pretreatment with dms ...202032106443
[eukaryotic expression, protein purification and biological effects research of human cs1-fc fusion protein].signaling lymphocyte activation family 7 (slamf7/cs1) is a cell surface glycoprotein that is highly expressed in multiple myeloma cells. cs1 is a sensitive and specific biomarker for multiple myeloma. car-t cell immunotherapy is a new method for the treatment of multiple myeloma. cs1 car-t cell immunotherapy has good effect on relapsed refractory multiple myeloma. to detect the expression efficiency of cs1 car on cs1 car-t cells and to find an auxiliary means to car-t cell immunotherapy, we prep ...202032072787
data for engineering lipid metabolism of chinese hamster ovary (cho) cells for enhanced recombinant protein production.the data presented in this article relates to the manuscript entitled 'engineering of chinese hamster ovary cell lipid metabolism results in an expanded er and enhanced recombinant biotherapeutic protein production', published in the journal metabolic engineering [1]. in the article here, we present data examining the overexpression of the lipid metabolism modifying genes scd1 and srebf1 in cho cells by densitometry of western blots and by using mass spectrometry to investigate the impact on spe ...202032071989
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 ...202032044516
slowing of the time course of acidification decreases the acid-sensing ion channel 1a current amplitude and modulates action potential firing in neurons.acid-sensing ion channels (asics) are h+-activated neuronal na+ channels. they are involved in fear behavior, learning, neurodegeneration after ischemic stroke and in pain sensation. asic activation has so far been studied only with fast ph changes, although the ph changes associated with many roles of asics are slow. it is currently not known whether slow ph changes can open asics at all. here, we investigated to which extent slow ph changes can activate asic1a channels and induce action potent ...202032180707
multi-omics reveals impact of cysteine feed concentration and resulting redox imbalance on cellular energy metabolism and specific productivity in cho cell bioprocessing.chinese hamster ovary (cho) cells are currently the primary host cell lines used in biotherapeutic manufacturing of monoclonal antibodies (mabs) and other biopharmaceuticals. cellular energy metabolism and endoplasmic reticulum (er) stress are known to greatly impact cell growth, viability, and specific productivity of a biotherapeutic; but the molecular mechanisms are not fully understood. the authors previously employed multi-omics profiling to investigate the impact of a reduction in cysteine ...202032170810
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 ...202032053646
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 ...202031990124
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 ...202032044029
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 ...202032041782
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 ...202031987841
altered gene expression in cho cells following polyamine starvation.to investigate the impact of polyamine deprivation on the transcriptome of cho cells results: polyamines play a central but poorly-understood role in cell proliferation. most studies to date have utilised chemical inhibitors to probe polyamine function. here we exploit the fact that cho cells grown in serum-free medium have an absolute requirement for putrescine supplementation due to their deficiency in activity of the enzyme arginase. a gene expression microarray (affymetrix) analysis of cho-k ...202032078082
improvement of intracellular traffic system by overexpression of kdel receptor 1 in antibody-producing cho cells.the localization of soluble endoplasmic reticulum (er) chaperones in the cell organelle is mediated by the c-terminal kdel (lysine, aspartic acid, glutamic acid and leucine) motif. this motif is recognized by the kdel receptor, a seven-transmembrane protein that cycles between the er and cis-golgi to capture missorted kdel chaperones from post-er compartments in a ph-dependent manner. the kdel receptor's target chaperones have a substantial role in protein folding and assembly. in this study, th ...202032073237
the use of catechins in chinese hamster ovary cell media for the improvement of monoclonal antibody yields and a reduction of acidic species.catechin compounds have potential benefits for recombinant monoclonal antibody (mab) production as chemical additives in cell culture media. in this study, four catechin compounds catechin (cat), epicatechin (ec), gallocatechin-gallate (gcg), and epigallocatechin-gallate (egcg) were added to cell culture media (at 50 μm) and their effects on the recombinant chinese hamster ovary (cho) cell culture, specific productivity, and mab quality were assessed. the results indicate that the improvement of ...202032067358
neutralizing antibody against osteopontin attenuates non-alcoholic steatohepatitis in mice.previously, we reported that an extracellular matrix protein, osteopontin (opn), is involved in various autoimmune diseases using a neutralizing polyclonal antibody against opn generated in rabbits. however, the antibody cannot be used for long-term mouse models of chronic inflammatory disease because of the induction of antibodies against anti-opn rabbit igg. in this study, we generated a new antibody, anti-mouse opn mouse igg (35b6). 35b6 inhibited the cell adhesion of mouse and human opn to c ...202032062834
understanding the effect of high gas entrance velocity on chinese hamster ovary (cho) cell culture performance and its implications on bioreactor scale-up and sparger design.there are three main potential sources for cell shear damage existing in stirred tank bioreactors. one is the potential high energy dissipation in the immediate impeller zones; another from small gas bubble burst; and third is from high gas entrance velocity (gev) emitting from the sparger. while the first two have been thoroughly addressed for the scale-up of chinese hamster ovary (cho) cell culture knowing that a wide tolerable agitation range with non-damaging energy dissipation exists and th ...202032086806
optimized crispr/cas9 strategy for homology-directed multiple targeted integration of transgenes in cho cells.site-specific integration has emerged as a promising strategy for precise chinese hamster ovary (cho) cell line engineering and predictable cell line development (cld). crispr/cas9 with the homology-directed repair (hdr) pathway enables precise integration of transgenes into target genomic sites. however, inherent recalcitrance to hdr-mediated targeted integration (ti) of transgenes results in low targeting efficiency, thus requiring a selection process to find a targeted integrant in cho cells. ...202032086804
identification and functional characterization of a novel splicing variant in the f8 coagulation gene causing severe hemophilia a.we have identified a synonymous f8 variation in a severe hemophilia a (ha) patient who developed inhibitors following factor viii (fviii) prophylaxis. the unreported c.6273 g > a variant targets the consensus splicing site of exon 21.202032078252
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 ...202031965756
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 ...202031964705
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 ...202031963132
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 ...202032039452
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 ...202031962234
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 ...202031981620
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 ...202032038177
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 ...202032035699
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 ...202032031796
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 ...202032023047
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 ...202032019132
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 ...202032012447
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 ...202031975519
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 ...202032005658
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 ...202031952136
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 ...202031949176
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 ...202031945727
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 ...202031957968
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 ...202031956990
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 ...202032001734
what cho is made of: variations in the biomass composition of chinese hamster ovary cell lines.cell line-specific, genome-scale metabolic models enable rigorous and systematic in silico investigation of cellular metabolism. such models have recently become available for chinese hamster ovary (cho) cells. however, a key ingredient, namely an experimentally validated biomass function that summarizes the cellular composition, was so far missing. here, we close this gap by providing extensive experimental data on the biomass composition of 13 parental and producer cho cell lines under various ...202032619503
improving chinese hamster ovary host cell protein elisa using ella®: an automated microfluidic platform.chinese hamster ovary (cho) cells are a mammalian cell line used in the production of therapeutic proteins. host cell proteins (hcps) are process-related impurities that are derived from the host cell expression system. during biopharmaceutical drug development, removal of hcps is required. enzyme-linked immunosorbent assay (elisa) is a common technique to quantitate hcps, but is a labor-intensive process that takes up to 7 h. ella® is an automated instrument that utilizes microfluidics and glas ...202032615786
directed evolution approach to enhance efficiency and speed of outgrowth during single cell subcloning of chinese hamster ovary cells.chinese hamster ovary (cho) cells are the working horse of the pharmaceutical industry. to obtain high producing cell clones and to satisfy regulatory requirements single cell cloning is a necessary step in cell line development. however, it is also a tedious, labor intensive and expensive process. here we show an easy way to enhance subclonability using subcloning by single cell sorting itself as the selection pressure, resulting in improved subcloning performance of three different host cell l ...202032612755
in situ fucosylation of the wnt co-receptor lrp6 increases its endocytosis and reduces wnt/β-catenin signaling.wnt/β-catenin signaling regulates critical, context-dependent transcription in numerous physiological events. among the well-documented mechanisms affecting wnt/β-catenin activity, modification of n-glycans by l-fucose is the newest and the least understood. using a combination of chinese hamster ovary cell mutants with different fucosylation levels and cell-surface fucose editing (in situ fucosylation [isf]), we report that α(1-3)-fucosylation of n-acetylglucosamine (glcnac) in the galβ(1-4)-gl ...202032649905
lc-ms/ms-based quantitative proteomic and phosphoproteomic analysis of cho-k1 cells adapted to growth in glutamine-free media.this study aims to provide insights into the molecular mechanisms underlying adaptation of cho-k1 cells to growth in glutamine-free media and potentially identifying critical signalling proteins and pathways involved in this phenotype.202032648187
s-adenosylmethionine and methylthioadenosine boost cellular productivities of antibody forming chinese hamster ovary cells.the improvement of cell specific productivities for the formation of therapeutic proteins is an important step towards intensified production processes. among others, the induction of the desired production phenotype via proper media additives is a feasible solution provided that said compounds adequately trigger metabolic and regulatory programs inside the cells. in this study, s-(5'-adenosyl)- l-methionine (sam) and 5'-deoxy-5'-(methylthio)adenosine (mta) were found to stimulate cell specific ...202032644191
media supplementation for targeted manipulation of monoclonal antibody galactosylation and fucosylation.monoclonal antibodies are critically important biologics as the largest class of molecules used to treat cancers, rheumatoid arthritis, and other chronic diseases. antibody glycosylation is a critical quality attribute that has ramifications for patient safety and physiological efficacy-one that can be modified by such factors as media formulation and process conditions during production. using a design-of-experiments approach, we examined the effect of 2-f-peracetyl fucose (2fp), uridine, and g ...202032662879
random epigenetic modulation of cho cells by repeated knockdown of dna methyltransferases increases population diversity and enables sorting of cells with higher production capacities.chinese hamster ovary (cho) cells produce a large share of today's biopharmaceuticals. still, the generation of satisfactory producer cell lines is a tedious undertaking. recently, it was found that cho cells, when exposed to new environmental conditions, modify their epigenome, suggesting that cells adapt their gene expression pattern to handle new challenges. the major aim of the present study was to employ artificially induced, random changes in the dna-methylation pattern of cho cells to div ...202032662873
characterization of glutathione proteome in cho cells and its relationship with productivity and cholesterol synthesis.glutathione (gsh) plays a central role in the redox balance maintenance in mammalian cells. previous studies of industrial chinese hamster ovary cell lines have demonstrated a relationship between gsh metabolism and clone productivity. however, a thorough investigation is required to understand this relationship and potentially highlight new targets for cell engineering. in this study, we have modulated the gsh intracellular content of an industrial cell line under bioprocess conditions to furth ...202032662871
immunoglobulin fragment f(ab')2 against rbd potently neutralizes sars-cov-2 in vitro.covid-19, which is caused by the emerging human coronavirus sars-cov-2, has become a global pandemic that poses a serious threat to human health. to date, no vaccines or specific antiviral drugs have been approved for the treatment of this disease in clinic. herein, therapeutic antibodies for sars-cov-2 were obtained from hyperimmune equine plasma. first, a recombinant sars-cov-2 spike protein receptor-binding domain (rbd) was obtained in gram-level quantities through high-cell density fermentat ...202032659292
investigation into the impact of tyrosine on the product formation and quality attributes of mabs in rcho cell cultures.tyrosine (tyr) is crucial to the maintenance of the monoclonal antibody (mab) titers and quality attributes in fed-batch cultures of recombinant chinese hamster ovary (rcho) cells. however, the relation between tyrosine and these aspects is not yet fully defined. in order to further elucidate such a relation, two groups of fed-batch experiments with high tyrosine (h-t) or low tyrosine (l-t) additions producing an igg1 monoclonal antibody against cd20 were implemented to investigate the intracell ...202032577803
quinoline-triazole half-sandwich iridium(iii) complexes: synthesis, antiplasmodial activity and preliminary transfer hydrogenation studies.iridium(iii) half-sandwich complexes containing 7-chloroquinoline-1,2,3-triazole hybrid ligands were synthesised and their inhibitory activities evaluated against the plasmodium falciparum malaria parasite. supporting computational analysis revealed that metal coordination to the quinoline nitrogen occurs first, forming a kinetic product that, upon heating over time, forms a more stable cyclometallated thermodynamic product. single crystal x-ray diffraction confirmed the proposed molecular struc ...202032697227
low density lipoprotein receptor variants in the beta-propeller subdomain and their functional impact.background: pathogenic variants in the low density lipoprotein receptor gene are associated with familial hypercholesterolemia. some of these variants can result in incorrect folding of the ldlr protein, which is then accumulated inside the cell and cannot fulfill its function to internalize ldl particles. we analyzed the functional impact of 10 ldlr variants localized in the beta-propeller of epidermal growth factor precursor homology domain. the experimental part of the work was complemented b ...202032695144
tailoring translational strength using kozak sequence variants improves bispecific antibody assembly and reduces product-related impurities in cho cells.optimal production of bispecific antibodies (bsab) requires efficient and tailored co-expression and assembly of two distinct heavy and two distinct light chains. here, we describe a novel technology to modulate the translational strength of antibody chains via kozak sequence variants to produce bsab in a single cell line. in this study, we designed and screened a large kozak sequence library to identify 10 independent variants that can modulate protein expression levels from approximately 0.2 t ...202032246763
clinical immunogenicity risk assessment for a fusion protein.this document highlights some relevant factors in the assessment of immunogenicity risk of fusion protein therapeutics. our aim is to highlight specific risks associated with this type of molecule, while also aligning with general risk assessment factors, through a hypothetical case study, where the therapeutic molecule of interest is a receptor-fc fusion protein (rff) expressed within a typical manufacturing process in chinese hamster ovary cells (cho). given that the components are comprised o ...202032246297
reduction of charge variants by cho cell culture process optimization.over the past decade, global interest in the development of therapeutic monoclonal antibodies (mabs) has risen rapidly. as therapeutic agents, antibodies have shown marked efficacy in combatting a range of cancers and immune diseases with high target specificity and low toxicity (carla lucia et al. in plos one 6:e24071, 2011; donaghy in mabs 8:659-671, 2016; nasiri et al. in j cell physiol 9:6441-6457, 2018; teo et al. in cancer immunol immunother 61:2295-2309, 2012). recent advances in cell cul ...202032236800
a role of heparan sulphate proteoglycan in the cellular uptake of lipocalins ß-lactoglobulin and allergen fel d 4.lipocalins, small extracellular hydrophobic molecule carriers, can be internalized by a variety of different cells. however, to date receptors have only been identified for human lipocalins. here, we specifically investigated uptake mechanisms for lipocalins ß-lactoglobulin and fel d 4 in hela and chinese hamster ovary (cho) cells. we provide evidence that cell surface heparan sulphate proteoglycan is essential for internalization of these lipocalins. in hela cells, lipocalin uptake was inhibite ...202032229688
improved identification of host cell proteins in a protein biopharmaceutical by lc-ms/ms using the proteominer™ enrichment kit.host cell proteins (hcps) in biotherapeutics can be identified by the use of enzymatic digestion and lc-ms/ms analysis. however, the major challenge is that hcps are often present at very low levels in relation to the protein drug (low ppm-levels). in this study, the proteominer™ enrichment kit (bio-rad) was evaluated as a strategy to enable identification of hcps by lc-ms/ms by enrichment of low-abundant hcps and a simultaneous depletion of the high-abundant product protein. a recombinant prote ...202032229402
immunogenic and inflammatory responses to citrullinated proteins are enhanced following modification with malondialdehyde-acetaldehyde adducts.malondialdehyde-acetaldehyde adducts (maa) act as potent immune adjuvants and co-localize with citrullinated antigens in tissues effected by rheumatoid arthritis (ra). we sought to examine the role of maa-adducts in promoting ra-related autoimmunity and inflammation.202032224441
nutrient supplementation strategy improves cell concentration and longevity, monoclonal antibody production and lactate metabolism of chinese hamster ovary cells.a careful selection of culture mediums and feeds has become necessary to maximize yields of recombinant proteins during bioprocesses of mammalian cells. supplements contain a variety of concentrate nutrients, and their beneficial effects vary according to recombinant cell lines. in this study, the effects of powerfeed a on growth kinetics, productivity and cellular metabolism were evaluated for two chinese hamster ovary cell lines producing a monoclonal antibody in a batch culture. supplemented ...202032223359
a metabolic network-based approach for developing feeding strategies for cho cells to increase monoclonal antibody production.chinese hamster ovary (cho) cells are the main workhorse in the biopharmaceutical industry for the production of recombinant proteins, such as monoclonal antibodies. to date, a variety of metabolic engineering approaches have been used to improve the productivity of cho cells. while genetic manipulations are potentially laborious in mammalian cells, rational design of cho cell culture medium or efficient fed-batch strategies are more popular approaches for bioprocess optimization. in this study, ...202032211960
a proteomic approach towards understanding crypoprotective action of me2so on the cho cell proteome.chinese hamster ovary (cho) cell lines are the most widely used in vitro cells for research and production of recombinant proteins such as rhgh, tpa, and erythropoietin. we aimed to investigate changes in protein profiles after cryopreservation using 2d-dige maldi-tof ms and network pathway analysis. the proteome changes that occur in cho cells between freshly prepared cells and cryopreserved cells with and without me2so were compared to determine the key proteins and pathways altered during rec ...202032259523
comparison of a novel homogeneous cyclic amp assay and a luciferase assay for measuring stimulating thyrotropin-receptor autoantibodies.stimulating thyrotropin-receptor antibodies (tsab) cause graves' disease (gd). we tested a novel homogeneous fluorescent 3',5' cyclic adenine monophosphate (camp) assay for the detection of tsab in a bioassay.202032257955
antiarrhythmic mechanisms of chinese herbal medicine dingji fumai decoction.dingji fumai decoction (dfd) is used to treat ventricular arrhythmia, and it has provided a very good curative effect. however, its cellular electrophysiological mechanism is unknown.202032256664
antibody-drug conjugates using mouse-canine chimeric anti-dog podoplanin antibody exerts antitumor activity in a mouse xenograft model.antibody-drug conjugates (adcs), which consist of a monoclonal antibody (mab), a linker, and a payload, can deliver a drug to cancer tissues. we previously produced an anti-dog podoplanin (dpdpn) mab, pmab-38, which reacts with dpdpn-expressing canine melanomas and squamous cell carcinomas (sccs), but not with dpdpn-expressing canine type i alveolar cells or lymphatic endothelial cells, indicating that pmab-38 possesses cancer specificity. in this study, we developed an adc, p38b-dm1, using the ...202032182186
cell function profiling to assess clone stability.in cell line development the identification of stable chinese hamster ovary cells for production is a critical but onerous task. the stability trial focus upon high-level attributes can mask profound underlying cellular changes, leading to unstable clones mistakenly being chosen for production. the challenge is to assay underlying cell pathways and subsystems without pushing up cell line development costs. chemstress® cell function profiling is a simple, multiwell plate-based assay that uses a p ...202032181887
hydrocyclones as cell retention device for cho perfusion processes in single-use bioreactors.in this study, a hydrocyclone (hc) especially designed for mammalian cell separation was applied for the separation of chinese hamster ovary cells. the effect of key features on the separation efficiency, such as type of pumphead in the peristaltic feed pump, use of an auxiliary pump to control the perfusate flow rate, and tubing size in the recirculation loop were evaluated in batch separation tests. based on these preliminary batch tests, the hc was then integrated to 50-l disposable bioreacto ...202032181883
evaluation of glycoprotein e subunit and live attenuated varicella-zoster virus vaccines formulated with a single-strand rna-based adjuvant.varicella-zoster virus (vzv), a human alphaherpesvirus 3, elicits both chickenpox and shingles and/or postherpetic neuralgia. a live attenuated vaccine (lav) and glycoprotein e (ge) subunit vaccine were developed to prevent vzv-induced diseases. we recently reported that single-strand rna (ssrna) based on the intergenic region of the internal ribosome entry site of cricket paralysis virus (crpv) is an effective adjuvant for protein-based and virus-like particle-based vaccines. here, chinese hams ...202032167678
review of the current methods of chinese hamster ovary (cho) cells cultivation for production of therapeutic protein.most of clinical approved protein-based drugs or under in clinical trial have a profound impact in the treatment of critical diseases. the mammalian eukaryotic cells culture approaches, particularly the cho (chinese hamster ovary) cells are mainly used in the biopharmaceutical industry for the mass-production of therapeutic protein. recent advances in cho cell bioprocessing to yield recombinant proteins and monoclonal antibodies have enabled the expression of quality protein. the developments of ...202032164511
effects of titanium dioxide nanoparticles on the hprt gene mutations in v79 hamster cells.the genotoxicity of anatase/rutile tio2 nanoparticles (tio2 nps, nm105 at 3, 15 and 75 µg/cm2) was assessed with the mammalian in-vitro hypoxanthine guanine phosphoribosyl transferase (hprt) gene mutation test in chinese hamster lung (v79) fibroblasts after 24 h exposure. two dispersion procedures giving different size distribution and dispersion stability were used to investigate whether the effects of tio2 nps depend on the state of agglomeration. tio2 nps were fully characterised in the previ ...202032150818
comparison of the pharmacological profiles of arginine vasopressin and oxytocin analogs at marmoset, macaque, and human vasopressin 1a receptor.arginine vasopressin (avp) and oxytocin (ot) are nonapeptides that bind to g-protein coupled receptors and influence social behaviors. consensus mammalian avp and ot (leu8-ot) sequences are highly conserved. in marmosets, an amino acid change in the 8th position of the peptide (pro8-ot) exhibits unique structural and functional properties. there is ∼85 % structural homology between the ot receptor (otr) and vasopressin 1a receptor (v1ar) resulting in significant cross-reactivity between the liga ...202032145592
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 ...202032058129
influence of polymer charge on the localization and dark- and photo-induced toxicity of a potential type i photosensitizer in cancer cell models.a current trend within photo-dynamic therapy (pdt) is the development of molecular systems targeting hypoxic tumors. thus, type i pdt sensitizers could here overcome traditional type ii molecular systems that rely on the photo-initiated production of toxic singlet oxygen. here, we investigate the cell localization properties and toxicity of two polymeric anthracene-based fluorescent probes (neutral ant-phea and cationic ant-pim). the cell death and dna damage of chinese hamster ovary cancer cell ...202032138280
enhancement of sialylation in rigg in glyco-engineered chinese hamster ovary cells.since about 70% of commercial biopharmaceutical products have been produced in chinese hamster ovary (cho) cells, this cell line is undeniably a workhorse for biopharmaceuticals production. meanwhile, sialic acid terminals were reported to affect anti-inflammatory activity, antibody-dependent cellular cytotoxicity efficacy of igg antibodies. taking these findings together, we aimed to establish cho cell lines that highly produce sialic acid terminals by overexpressing two n-acetylneuraminic acid ...202032125558
1-methylpyrene induces chromosome loss and mitotic apparatus damage in a chinese hamster v79-derived cell line expressing both human cyp1a2 and sulfotransferase 1a1.1-methylpyrene (1-mp) is a ubiquitous environmental pollutant and rodent carcinogen. its mutagenic activity depends on sequential activation by various cyp and sulfotransferase (sult) enzymes. previously we have observed induction of micronuclei and mitotic arrest by 1-mp in a chinese hamster (v79)-derived cell line expressing both human cyp1a2 and sult1a1 (v79-hcyp1a2-hsult1a1), however, the mode of chromosome damage and the involvement of mitotic tubulin structures have not been clarified. in ...202033035519
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 ...202031956982
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