TitleAbstractYear(sorted ascending)
development of a chemically defined platform fed-batch culture media for monoclonal antibody-producing cho cell lines with optimized choline content.a chemically defined platform basal medium and feed media were developed using a single chinese hamster ovary (cho) cell line that produces a monoclonal antibody (mab). cell line a, which showed a peak viable cell density of 5.9 × 106 cells/ml and a final mab titer of 0.5 g/l in batch culture, was selected for the platform media development. stoichiometrically balanced feed media were developed using glucose as an indicator of cell metabolism to determine the feed rates of all other nutrients. a ...201829322349
effectiveness of aqueous and hydroalcoholic extracts of acanthospermum australe (loefl.) kuntze against diarrhea-inducing bacteria.leaves and roots of acanthospermum australe (asteraceae) have been used in brazilian folk medicine for the treatment of various ailments including diarrhea, skin diseases, blennorrhagia, dyspepsia, parasitic worms and malaria. the aim of study was to characterize the chemical profiles of the aqueous and hydroalcoholic extracts of leaves and roots of a. australe, and to evaluate their antimicrobial activities against diarrhea-inducing bacteria (enterococcus faecalis, shigella dysenteriae and yers ...201829319752
rna-seq highlights high clonal variation in monoclonal antibody producing cho cells.the development of next-generation sequencing technologies has opened new opportunities to better characterize complex eukaryotic cells. chinese hamster ovary (cho) cells play a primary role in therapeutic protein production, with currently five of the top ten blockbuster drugs produced in cho. however, engineering superior cho cells with improved production features has had limited success to date and cell lines are still developed through the generation and screening of large strain pools. her ...201829316330
glycoengineering in cho cells: advances in systems biology.for several decades, glycoprotein biologics have been successfully produced from chinese hamster ovary (cho) cells. the therapeutic efficacy and potency of glycoprotein biologics are often dictated by their post translational modifications, particularly glycosylation, which unlike protein synthesis, is a non-templated process. consequently, both native and recombinant glycoprotein production generate heterogeneous mixtures containing variable amounts of different glycoforms. stability, potency, ...201829316325
methods for using small non-coding rnas to improve recombinant protein expression in mammalian cells.the ability to produce recombinant proteins by utilizing different "cell factories" revolutionized the biotherapeutic and pharmaceutical industry. chinese hamster ovary (cho) cells are the dominant industrial producer, especially for antibodies. human embryonic kidney cells (hek), while not being as widely used as cho cells, are used where cho cells are unable to meet the needs for expression, such as growth factors. therefore, improving recombinant protein expression from mammalian cells is a p ...201829315258
evaluation of different vector design strategies for the expression of recombinant monoclonal antibody in cho cells.monoclonal antibodies (mabs) have emerged as the most promising category of recombinant proteins due to their high efficiency for the treatment of a wide range of human diseases. the complex nature of mabs creates a great deal of challenges in both upstream and downstream manufacturing processes. proportional expression and correct folding and assembly of the light chain and heavy chain are required for efficient production of the mabs. in this regard, expression vector design has proven to have ...201829313429
ecotoxicological potential of the biocides terbutryn, octhilinone and methylisothiazolinone: underestimated risk from biocidal pathways?the use of biocides by industry, agriculture and households increased throughout the last two decades. many new applications with known substances enriched the variety of biocidal pollution sources for the aquatic environment. while agriculture was the major source for a long time, leaching from building facades and preservation of personal care and cleaning products was identified as new sources in the last few years. with the different usage forms of biocidal products the complexity of legisla ...201829306832
antidepressant drug paroxetine blocks the open pore of kv3.1 potassium patients with epilepsy, depression is a common comorbidity but difficult to be treated because many antidepressants cause pro-convulsive effects. thus, it is important to identify the risk of seizures associated with antidepressants. to determine whether paroxetine, a very potent selective serotonin reuptake inhibitor (ssri), interacts with ion channels that modulate neuronal excitability, we examined the effects of paroxetine on kv3.1 potassium channels, which contribute to highfrequency fir ...201829302214
in vitro binding and receptor-mediated activity of terlipressin at vasopressin receptors v1 and v2.terlipressin, a synthetic, systemic vasoconstrictor with selective activity at vasopressin-1 (v1) receptors, is a pro-drug for the endogenous/natural porcine hormone [lys8]-vasopressin (lvp). we investigated binding and receptor-mediated cellular activities of terlipressin, lvp, and endogenous human hormone [arg8]-vasopressin (avp) at v1 and vasopressin-2 (v2) receptors. cell membrane homogenates of chinese hamster ovary cells expressing human v1 and v2 receptors were used in competitive binding ...201829302194
blood coagulation factor x exerts differential effects on adenovirus entry into human has been proposed that blood coagulation factors, principally factor x (fx), enhance the uptake of human adenovirus type 5 (ad5) into cultured epithelial cells by bridging the viral hexon capsid protein and cell-surface heparan sulphate proteoglycans (hspgs). we studied the effects of fx on ad transduction of lymphoid cell lines (nk92mi, a natural killer cell line; daudi, a b-cell line and jurkat, a t-cell line) as well as primary peripheral blood lymphocytes (pbl) and hela epithelial cells u ...201829301346
in vitro genotoxicity assessment of dinitroaniline herbicides pendimethalin and trifluralin.pendimethalin and trifluralin are widely used dinitroaniline herbicides. both compounds can be found as residue levels in agricultural products. this study was conducted in order to provide necessary information for the risk assessment of pendimethalin and trifluralin. in this study, reactive oxygen species (ros) levels were measured to examine the potential of both compounds to induce oxidative damage in chinese hamster lung fibroblast (v79) cells. also, the genotoxic effects of pendimethalin a ...201829374591
ring finger protein 145 (rnf145) is a ubiquitin ligase for sterol-induced degradation of hmg-coa reductase.cholesterol biosynthesis is tightly regulated in the cell. for example, high sterol concentrations can stimulate the degradation of the rate-limiting cholesterol biosynthesis enzyme 3-hydroxy-3-methylglutaryl-coenzyme a reductase (hmg-coa reductase, hmgcr). hmgcr is broken down by endoplasmic reticulum (er) membrane-associated protein complexes consisting of insulin-induced genes (insigs) and the e3 ubiquitin ligase gp78. here, we found that hmgcr degradation is partially blunted in chinese hams ...201829374057
design and evaluation of a novel flavonoid-based radioprotective agent utilizing monoglucosyl this study, three novel flavonoid composite materials, created by combining an aglycone [quercetin (que), hesperetin (hes) or naringenin (nar)] with monoglucosyl rutin (mgr), were designed to test for improved radioprotectivity compared with that provided by administration of mgr alone. aglycone in the mgr-composite state was highly soluble in water, compared with aglycone alone dissolved in dimethyl sulfoxide or distilled water. the antioxidant activity of the three flavonoid composites was ...201829373678
investigation into the mechanism of homo- and heterodimerization of angiotensin-converting enzyme.angiotensin-converting enzyme (ace) plays a central role in the renin-angiotensin system (ras), which is primarily responsible for blood pressure homeostasis. studies have shown that ace inhibitors yield cardiovascular benefits that cannot be entirely attributed to the inhibition of ace catalytic activity. it is possible that these benefits are due to interactions between ace and ras receptors that mediate the protective arm of the ras, such as the angiotensin-ii receptor type 2 (at2r) and the r ...201829371233
development of a novel 99m tc-labelled small molecular antagonist for cxcr4 positive tumor imaging.the chemokine receptor 4 (cxcr4) has been an attractive molecular target for tumor imaging, since it is overexpressed in many tumor types and involved in tumor progression and metastasis. the purpose of this study is to examine the cxcr4 targeting properties of 99m tc-labeled amd3465, a small molecule antagonist of cxcr4. 99m tc-amd3465 was prepared in high yield (>95%) and stable in mice serum at least for 4 h. in vitro cell binding experiments were performed with cho (chinese hamster ovary), m ...201829370457
opp labelling enables total protein synthesis quantification in cho production cell lines at the single-cell level.accurate measurement of global and specific protein synthesis rates becomes increasingly important, especially in the context of biotechnological applications such as process modelling or selection of production cell clones. while quantification of total protein translation across whole cell populations is easily achieved, methods that are capable of tracking population dynamics at the single-cell level are still lacking. to address this need, we applied o-propargyl-puromycin (opp) labelling to ...201829369524
enhanced production of anti-pd1 antibody in cho cells through transient co-transfection with anti-apoptotic genes bcl-x l and mcl-1.apoptosis has a negative impact on the cell survival state during cell cultivation. to optimize mammalian cell culture for production of biopharmaceuticals, one of the important approaches is to extend cell life through over-expression of anti-apoptotic genes. here, we reported a cost-effective process to enhance cell survival and production of an antibody through transient co-transfection with anti-apoptotic genes bcl-x l or mcl-1 in chinese hamster ovary (cho) cells with polyethylenimine (pei) ...201829368032
development of novel oxazolo[5,4-d]pyrimidines as competitive cb2 neutral antagonists based on scaffold hopping.a series of novel oxazolo[5,4-d]pyrimidines was designed via a scaffold hopping strategy and synthesized through a newly developed approach. all these compounds were evaluated for their biological activity toward cb1/cb2 cannabinoid receptors, their metabolic stability in mice liver microsomes and their cytotoxicity against several cell lines. eight compounds have been identified as cb2 ligands with ki values less than 1 μm. it is noteworthy that 2-(2-chlorophenyl)-5-methyl-7-(4-methylpiperazin- ...201829360044
using titer and titer normalized to confluence are complementary strategies for obtaining chinese hamster ovary cell lines with high volumetric productivity of etanercept.the selection of clonally-derived chinese hamster ovary (cho) cell lines with the highest production rate of recombinant glycoproteins remains a big challenge during early stages of cell line development. different strategies using either product titer or product titer normalized to cell number are being used to assess suspension-adapted clones when grown statically in microtiter plates. however, no reported study so far has performed a direct head-to-head comparison of these two early reporters ...201829359860
the genotoxicity of an aqueous extract of gyejibokryeong-hwan.gyejibokryeong-hwan (guizhi fuling wan in china), a mixture of five herbal plants, is a well-known treatment for renal diseases including those associated with climacteric syndrome. however, the genotoxicity of gyejibokryeong-hwan has not yet been well established.201829357857
ampulexins: a new family of peptides in venom of the emerald jewel wasp, ampulex compressa.the parasitoid wasp ampulex compressa injects venom directly into the brain and subesophageal ganglion of the cockroach periplaneta americana, inducing a seven to ten day lethargy termed hypokinesia. hypokinesia presents as a significant reduction in both escape response and spontaneous walking. we examined aminergic and peptidergic components of milked venom with hplc and maldi-tof mass spectrometry. hplc coupled with electrochemical detection confirmed presence of dopamine in milked venom, whi ...201829350905
amp and adenosine are both ligands for adenosine 2b receptor signaling.adenosine is considered the canonical ligand for the adenosine 2b receptor (a2br). a2br is upregulated following kidney ischemia augmenting post ischemic blood flow and limiting tubular injury. in this context the beneficial effect of a2br signaling has been attributed to an increase in the pericellular concentration of adenosine. however, following renal ischemia both kidney adenosine monophosphate (amp) and adenosine levels are substantially increased. using computational modeling and calcium ...201829191553
distinctive binding properties of the negative allosteric modulator, [3h]sb269,652, at recombinant dopamine d3 receptors.recently, employing radioligand displacement and functional coupling studies, we demonstrated that sb269,652 (n-[(1r,4r)-4-[2-(7-cyano-1,2,3,4-tetrahydroisoquinolin-2-yl)ethyl]cyclohexyl]-1h-indole-2-carboxamide) interacts in an atypical manner with dopamine d3 receptor displaying a unique profile reminiscent of a negative allosteric ligand. here, we characterized the binding of radiolabelled [3h]sb269,652 to human dopamine d3 receptor stably expressed in chinese hamster ovary cells. under satur ...201829223348
insights into the generation of monoclonal antibody acidic charge variants during chinese hamster ovary cell cultures.charge variation is one of the most important heterogeneities during monoclonal antibody (mab) manufacturing and this study presents insights into the generation of acidic charge variants during cell culture processes. since acidic variants generate both intracellularly and extracellularly, main charge fraction collected by weak cation exchange chromatography (wcx) was incubated in harvested cell supernatant (hcs) to simulate and investigate the extracellular process firstly. it is found that th ...201829238873
abin-1 negatively regulates μ-opioid receptor function.the μ-opioid receptor (mor) is a gi/o protein-coupled receptor that mediates analgesic, euphoric, and reward effects. using a bacterial two-hybrid screen, we reported that the carboxyl tail of the rat mor associates with a20-binding inhibitor of nuclear factor κb (abin-1). this interaction was confirmed by direct protein-protein binding and coimmunoprecipitation of mor and abin-1 proteins in cell lysates. saturation binding studies showed that abin-1 had no effect on mor binding. however, the in ...201829237725
haloacetic acid water disinfection byproducts affect pyruvate dehydrogenase activity and disrupt cellular metabolism.the disinfection of drinking water has been a major public health achievement. however, haloacetic acids (haas), generated as byproducts of water disinfection, are cytotoxic, genotoxic, mutagenic, carcinogenic, and teratogenic. previous studies of monohaa-induced genotoxicity and cell stress demonstrated that the toxicity was due to inhibition of glyceraldehyde-3-phosphate dehydrogenase (gapdh), leading to disruption of cellular metabolism and energy homeostasis. dihaas and trihaas are also prod ...201829261292
comparative study of the effects of phosphatidylcholine rich in dha and epa on alzheimer's disease and the possible mechanisms in cho-app/ps1 cells and samp8 mice.metabolic stress induced by a high-fat (hf) diet leads to cognitive dysfunction and aging. in the present study, chinese hamster ovary cells stably transfected with amyloid precursor protein (app) and presenilin 1 (ps1) (cho-app/ps1 cells) and samp8 mice fed with an hf diet were used to study the effects of docosahexaenoic acid (dha)-enriched phosphatidylcholine (dha-pc) and eicosapentaenoic acid (epa)-enriched phosphatidylcholine (epa-pc) on alzheimer's disease (ad) and the possible mechanisms ...201829292421
evaluation of reversed-phase nano liquid chromatography conditions by using reversed-phase thin layer chromatography based on hansen solubility parameters for the analysis of amphiphilic glycosylsphingolipid transformations.pulse chase analysis is often used in investigating dynamics of cellular substances. fluorescently labeled lactosyl sphingosine molecule is useful in chasing its transformation, however the analysis of such metabolites in attomole level is of extreme difficult due to the presence of large amount of endogenous amphiphilic molecules such as glycosphingolipids, sphingomyerin, and glycerophospholipids. nano lc suites for analyzing the attomole scale metabolites, therefore removal of endogenous subst ...201829290400
antigenotoxicity properties of copaifera multijuga oleoresin and its chemical marker, the diterpene (-)-copalic view of the biological activities and growing therapeutic interest in oleoresin obtained from copaifera multijuga, this study aimed to determine the genotoxic and antigenotoxic potential of this oleoresin (cmo) and its chemical marker, diterpene (-)-copalic acid (ca). the micronucleus (mn) assay in v79 cell cultures and the ames test were used for in vitro analyses, as well as mn and comet assays in swiss mice for in vivo analyses. the in vitro genotoxicity/mutagenicity results showed that ei ...201829286884
regulatory role of mir-18a to ccn2 by tgf-β1 signaling pathway in pulmonary injury induced by nano-sio2.this research is designed to investigate the regulatory effect of mir-18a to the target gene connective tissue growth factor (ctgf, or ccn2), by participating in tgf-β1 signaling pathway and explore the pathogenic mechanism of mir-18a in pulmonary injury induced by nano-sio2 based on our early study. mir-18a and expression of tgf-β1 in chinese hamster lung (chl) fibroblasts cells stimulated by supernatants of nr8383 cells exposed to 40 μg/ml nano-sio2 for 24 h demonstrated 1.58 ± 0.22-fold and 1 ...201829067610
z-505 hydrochloride ameliorates chemotherapy-induced anorexia in rodents via activation of the ghrelin receptor, ghsr1a.despite its therapeutic advantages, chemotherapy with anti-cancer drugs can cause adverse effects, including anorexia and weight loss. although most patients with cancer suffer from anorexia during chemotherapy, resulting in the need to suspend or cease treatment and thereby worsening prognosis, treatment options for anorexia remain limited. ghrelin is an orexigenic hormone that has been proposed to prevent anorexia. to investigate the potential of ghrelin receptor agonists, synthetic small-mole ...201829066414
functional characterization of the dual allatostatin-a receptors in mosquitoes.the neuropeptide allatostatin-a (asta) and its cognate receptors (astars) are involved in the modulation of feeding behavior, which in hematophagous insects includes the regulation of the disease vector-related behaviors, host seeking and blood feeding. in mosquitoes and other dipterans, there are two copies of astar, contrasting with the single copy found in other insects. in this study, we identified and cloned the dual astar system of two important disease vectors aedes aegypti and culex quin ...201829103918
chemical characterization and cytotoxic, genotoxic, and mutagenic properties of baccharis trinervis (lam, persoon) from colombia and brazil.baccharis trinervis (lam, persoon) leaves are used in the traditional medicine for the treatment of high fevers, edema, inflammation, sores and muscle cramps, snakebites and as antiseptic.201829100934
identification of a potent mar element from the human genome and assessment of its activity in stably transfected cho cells.low-level and unstable transgene expression are common issues using the cho cell expression system. matrix attachment regions (mars) enhance transgene expression levels, but additional research is needed to improve their function and to determine their mechanism of action. mar-6 from cho chromosomes actively mediates high and consistent gene expression. in this study, we compared the effects of two new mars and mar-6 on transgene expression in recombinant cho cells and found one potent mar eleme ...201829077269
proteomic analysis of cho cell lines producing high and low quantities of a recombinant antibody before and after selection with methotrexate.high levels of recombinant protein production in chinese hamster ovary (cho) cells can be achieved by amplification of transgenes using the dihydrofolate reductase/methotrexate (dhfr/mtx) system. with the aim to identify predictive markers enabling the preselection of suitable high producing clones we investigated the impact of mtx-based gene amplification on two cho cells lines producing different levels of a human monoclonal antibody by carrying out a comparative proteome analysis. the differe ...201829137976
dietary k+ and cl- independently regulate basolateral conductance in principal and intercalated cells of the collecting duct.the renal collecting duct contains two distinct cell types, principal and intercalated cells, expressing potassium kir4.1/5.1 (kcnj10/16) and chloride clc-k2 (clc-kb in humans) channels on their basolateral membrane, respectively. both channels are thought to play important roles in controlling systemic water-electrolyte balance and blood pressure. however, little is known about mechanisms regulating activity of kir4.1/5.1 and clc-k2/b. here, we employed patch clamp analysis at the single channe ...201829134279
using specific recombinant gonadotropins to induce spermatogenesis and spermiation in the european eel (anguilla anguilla).new specific european eel (anguilla anguilla) recombinant gonadotropins (aargths) produced in the ovarian cells of chinese hamsters (cho) were used to induce maturation in captive male eels. in the first experiment, five different hormonal treatments were assayed: one group was given a constant dose of recombinant european eel follicle-stimulating hormone (aarfsh; 4 μg/fish) for 9 weeks, and the second group received a constant dose of recombinant european eel luteinizing hormone (aarlh; 2 μg/fi ...201829120707
tolerability, response and outcome of high-risk neuroblastoma patients treated with long-term infusion of anti-gd2 antibody ch14.18/cho.immunotherapy with short term infusion (sti) of monoclonal anti-gd2 antibody (mab) ch14.18 (4 × 25 mg/m2/d; 8-20 h) in combination with cytokines and 13-cis retinoic acid (ra) prolonged survival in high-risk neuroblastoma (nb) patients. here, we investigated long-term infusion (lti) of ch14.18 produced in chinese hamster ovary cells (ch14.18/cho; 10 × 10 mg/m2; 24 h) in combination with subcutaneous (s.c.) interleukin-2 (il-2) in a single center program and report clinical response, toxicity and ...201829120699
combinations of chromosome transfer and genome editing for the development of cell/animal models of human disease and humanized animal models.chromosome transfer technology, including chromosome modification, enables the introduction of mb-sized or multiple genes to desired cells or animals. this technology has allowed innovative developments to be made for models of human disease and humanized animals, including down syndrome model mice and humanized transchromosomic (tc) immunoglobulin mice. genome editing techniques are developing rapidly, and permit modifications such as gene knockout and knockin to be performed in various cell li ...201829180645
types of cell death and apoptotic stages in chinese hamster ovary cells distinguished by raman spectroscopy.cell death is the ultimate cause of productivity loss in bioreactors that are used to produce therapeutic proteins. we investigated the ability of raman spectroscopy to detect the onset and types of cell death for chinese hamster ovary (cho) cells-the most widely used cell type for therapeutic protein production. raman spectroscopy was used to compare apoptotic, necrotic, autophagic, and control cho cells. several specific nucleic acid-, protein-, and lipid-associated marker bands within the 650 ...201829030978
kinetic characterization of bile salt transport by human ntcp (slc10a1).the transport of bile acids facilitated by ntcp is an important factor in establishing bile flow. in this study, we examine the kinetics associated with human ntcp-dependent transport of two quantitatively important bile acids comprising the human bile acid pool, chenodeoxycholic acid and glycine-chenodeoxycholate, and secondary bile salt, 3-sulfo-glycolithocholate of potential toxicological significance. the study employed human ntcp overexpressing chinese hamster ovary cells and results compar ...201829024779
ca2+ -dependent down-regulation of human histamine h1 receptors in chinese hamster ovary protein-coupled human histamine h1 receptors in chinese hamster ovary cells stimulated with histamine undergo clathrin-dependent endocytosis followed by proteasome/lysosome-mediated down-regulation. in this study, we evaluated the effects of a sustained increase in intracellular ca2+ concentrations induced by a receptor-bypassed stimulation with ionomycin, a ca2+ ionophore, on the endocytosis and down-regulation of h1 receptors in chinese hamster ovary cells. all cellular and cell-surface ...201829063596
role of 6-o-α-maltosyl-β-cyclodextrin in lysosomal cholesterol deprivation in npc1-deficient chinese hamster ovary cells.we aimed to investigate whether 6-o-α-maltosyl-β-cyclodextrin (mal-βcd) is incorporated into cells and lysosomes during the release of unesterified cholesterol in npc1-deficient chinese hamster ovary (cho) cells (npc1 ko cells) and cho-jp17 cells (jp17 cells). internalization of mal-βcd in cells and lysosomes and extracellular release of lysosomal unesterified cholesterol were demonstrated by lc/ms/ms and lc/ms, respectively. internalization of mal-βcd was greater in npc1 ko cells than in jp17 c ...201829172126
high frequency electroporation efficiency is under control of membrane capacitive charging and voltage potential relaxation.the study presents the proof of concept for a possibility to achieve a better electroporation in the mhz pulse repetition frequency (prf) region compared to the conventional low frequency protocols. the 200ns×10 pulses bursts of 10-14kv/cm have been used to permeabilize chinese hamster ovary (cho) cells in a wide range (1hz-1mhz) of prf. the permeabilization efficiency was evaluated using fluorescent dye assay (propidium iodide) and flow cytometry. it was determined that a threshold prf exists w ...201828922628
exploring cellular behavior under transient gene expression and its impact on mab productivity and fc-glycosylation.transient gene expression (tge) is a methodology employed in bioprocessing for the fast provision of recombinant protein material. mild hypothermia is often introduced to overcome the low yield typically achieved with tge and improve specific protein productivity. it is therefore of interest to examine the impact of mild hypothermic temperatures on both the yield and quality of transiently expressed proteins and the relationship to changes in cellular processes and metabolism. in this study, we ...201828921534
karyotype variation of cho host cell lines over time in culture characterized by chromosome counting and chromosome painting.genomic rearrangements are a common phenomenon in rapidly growing cell lines such as chinese hamster ovary (cho) cells, a feature that in the context of production of biologics may lead to cell line and product instability. few methods exist to assess such genome wide instability. here, we use the population distribution of chromosome numbers per cell as well as chromosome painting to quantify the karyotypic variation in several cho host cell lines. cho-s, cho-k1 8 mm glutamine, and cho-k1 cells ...201828921524
fed-batch cho cell culture for lab-scale antibody production.fed-batch culture is the most commonly used upstream process in industry today for recombinant monoclonal antibody production using chinese hamster ovary (cho) cells. developing and optimizing this process in the lab is crucial for establishing process knowledge, which enables rapid and predictable tech-transfer to manufacturing scale. in this chapter, we describe stepwise how to carry out fed-batch cho cell culture for lab-scale antibody production.201828921435
serological and molecular investigation of 2117-like vesiviruses in cats.vesivirus 2117 was first discovered as a contaminant in chinese hamster ovary (cho) cell cultures used for human drug production. similar vesiviruses (vevs) have been detected recently in dogs. in order to address the hypothesis that cats may also be exposed to 2117-like vevs, in this study, we screened 236 feline sera using an enzyme-linked immunosorbent assay (elisa) based on a recombinant vp1 protein from the canine vev bari/212/07/ita. igg antibodies against the 2117-like vev were detected i ...201829022118
evaluation of genotoxicity and subchronic toxicity of the standardized leaves infusion extract of copaifera malmei harms in experimental models.copaifera malmei harms (fabaceae), known mainly as óleo-mirim, is a native and endemic plant found in the states of mato grosso and goiás of brazil. the plant's leaves infusion is popularly used by riverine communities of the northern araguaia microregion, mato grosso, brazil, for the treatment of gastric ulcers and inflammatory diseases of the respiratory tract. the gastric antiulcer activity of the standardized leaves infusion extract of copaifera malmei (siecm) in rodents has been reported. t ...201828943446
magnetic field direction differentially impacts the growth of different cell types.magnetic resonance imaging (mri) machines have horizontal or upright static magnetic field (smf) of 0.1-3 t (tesla) at sites of patients and operators, but the biological effects of these smfs still remain elusive. we examined 12 different cell lines, including 5 human solid tumor cell lines, 2 human leukemia cell lines and 4 human non-cancer cell lines, as well as the chinese hamster ovary cell line. permanent magnets were used to provide 0.2-1 t smfs with different magnetic field directions. w ...201829621414
all-trans retinoic acid in combination with sodium butyrate enhances specific monoclonal antibody productivity in recombinant cho cell line.the effects of all-trans retinoic acid (ra) and sodium butyrate (nabu) on growth, viability and antibody production of two types of transfected chinese hamster ovary cell lines (cho-k1 and cho-s) were investigated using a batch mode cell culture. by adding 0.5 mm nabu in the cho-k1 cell culture, the cell specific productivity (qp) and antibody concentration increased by five- and threefold, respectively. the optimal concentration of ra was 100 nm which resulted in twofold increase in antibody pr ...201829619548
monoclonal antibody l1mab-13 detected human pd-l1 in lung cancers.programmed cell death ligand-1 (pd-l1) is a type i transmembrane glycoprotein expressed on antigen-presenting cells. it is also expressed in several tumor cells such as melanoma and lung cancer cells. a strong correlation has been reported between human pd-l1 (hpd-l1) expression in tumor cells and negative prognosis in cancer patients. here, a novel anti-hpd-l1 monoclonal antibody (mab) l1mab-13 (igg1, kappa) was produced using a cell-based immunization and screening (cbis) method. we investigat ...201829608408
multiple reaction monitoring targeted lc-ms analysis of potential cell death marker proteins for increased bioprocess control.the monitoring of protein biomarkers for the early prediction of cell stress and death is a valuable tool for process characterization and efficient biomanufacturing control. a representative set of six proteins, namely gpdh, prdx1, lgals1, cfl1, tagln2 and mdh, which were identified in a previous cho-k1 cell death model using discovery lc-mse was translated into a targeted liquid chromatography multiple reaction monitoring mass spectrometry (lc-mrm-ms) platform and verified. the universality of ...201829607450
mechanisms driving the lactate switch in chinese hamster ovary cells.the metabolism of chinese hamster ovary (cho) cells in a production environment has been extensively investigated. however, a key metabolic transition, the switch from lactate production to lactate consumption, remains enigmatic. though commonly observed in cho cultures, the mechanism(s) by which this metabolic shift is triggered is unknown. despite this, efforts to control the switch have emerged due to the association of lactate consumption with improved cell growth and productivity. this revi ...201829603726
toxic hypf-n oligomers selectively bind the plasma membrane to impair cell adhesion capability.the deposition of fibrillar protein aggregates in human organs is the hallmark of several pathological states, including highly debilitating neurodegenerative disorders and systemic amyloidoses. it is widely accepted that small oligomers arising as intermediates in the aggregation process, released by fibrils, or growing in secondary nucleation steps are the cytotoxic entities in protein-misfolding diseases, notably neurodegenerative conditions. increasing evidence indicates that cytotoxicity is ...201829590593
combination of temperature shift and hydrolysate addition regulates anti-ige monoclonal antibody charge heterogeneity in chinese hamster ovary cell fed-batch culture.charge heterogeneity has been broadly studied as a critical quality attribute during monoclonal antibody (mab) production that may subsequently affect product stability and biopotency. however, the charge variation distribution is poorly controlled, so methods of more effective control need to be explored. in this study, the combined effects of temperature shift (37-34, 37-32, or 37-30 °c) and hydrolysate addition (0.100 g/l) to culture feed on the charge heterogeneity of anti-ige mab were inves ...201829589263
an engineered complement factor h construct for treatment of c3 glomerulopathy.background c3 glomerulopathy (c3g) is associated with dysregulation of the alternative pathway of complement activation, and treatment options for c3g remain limited. complement factor h (fh) is a potent regulator of the alternative pathway and might offer a solution, but the mass and complexity of fh makes generation of full-length fh far from trivial. we previously generated a mini-fh construct, with fh short consensus repeats 1-5 linked to repeats 18-20 (fh1-5^18-20), that was effective in ex ...201829588430
identification of high affinity her2 binding antibodies using cho fab surface display.discovery of monoclonal antibodies is most commonly performed using phage or yeast display but mammalian cells are used for production because of the complex antibody structure, including the multiple disulfide bonds and glycosylation, required for function. as this transition between host organisms is often accompanied by impaired binding, folding or expression, development pipelines include laborious plate-based screening or engineering strategies to adapt an antibody to mammalian expression. ...201829566240
mild hypothermia upregulates myc and xbp1s expression and improves anti-tnfα production in cho cells.chinese hamster ovary (cho) cells are the most frequently used host for commercial production of therapeutic proteins. however, their low protein productivity in culture is the main hurdle to overcome. mild hypothermia has been established as an effective strategy to enhance protein specific productivity, although the causes of such improvement still remain unclear. the self-regulation of global transcriptional regulatory factors, such as myc and xbp1s, seems to be involved in increased the reco ...201829566086
inhibition of the α-subunit of phosphoinositide 3-kinase in heart increases late sodium current and is arrhythmogenic.although inhibition of phosphoinositide 3-kinase (pi3k) is an emerging strategy in cancer therapy, we and others have reported that this action can also contribute to drug-induced qt prolongation and arrhythmias by increasing cardiac late sodium current (inal). previous studies in mice implicate the pi3k-α isoform in arrhythmia susceptibility. here, we have determined the effects of new anticancer drugs targeting specific pi3k isoforms on inal and action potentials (aps) in mouse cardiomyocytes ...201829563327
human umbilical cord blood serum-derived α-secretase: functional testing in alzheimer's disease mouse models.alzheimer's disease (ad) is an age-related disorder that affects cognition. our previous studies showed that the neuroprotective fragment of amyloid procurer protein (app) metabolite, soluble appα (sappα), interferes with β-site app-cleaving enzyme 1 (bace1, β-secretase) cleavage and reduces amyloid-β (aβ) generation. in an attempt to identify approaches to restore sappα levels, we found that human cord blood serum (cbs) significantly promotes sappα production compared with adult blood serum (ab ...201829560732
toxicological evaluation of 6'-sialyllactose (6'-sl) sodium salt.we performed a series of toxicity studies on the safety of 6'-sialyllactose (6'-sl) sodium salt as a food ingredient. 6'-sl sodium salt, up to a maximum dose of 5000 μg/plate, did not increase the number of revertant colonies in five strains of salmonella typhimurium in the presence or absence of s9 metabolic activation. a chromosomal aberration assay (using chinese hamster lung cells) found no clastogenic effects at any concentration of 6'-sl sodium salt in the presence or absence of s9 metabol ...201829555556
heterologous expression of three antigenic proteins from angiostrongylus cantonensis: es-7, lec-5, and 14-3-3 in mammalian cells.angiostrongylus cantonensis is a parasitic nematode and the main causative agent of human cerebral eosinophilic meningoencephalitis (eom). a definitive diagnosis of eom usually requires serologic or molecular analysis of the patient's clinical sample. currently, a 31 kda antigen is used in immunological tests for this purpose, however as a crude antigen preparation it may present cross-reactivity with other helminthic infections, especially echinococcosis. heterologous expression studies using p ...201829555232
chemical modification of protein a chromatography ligands with polyethylene glycol. ii: effects on resin robustness and process selectivity.we have proposed chemical modification of protein a (proa) chromatography ligands with polyethylene glycol (pegylation) as a strategy to increase the resin selectivity and robustness by providing the ligand with a steric repulsion barrier against non-specific binding. here, we report on robustness and selectivity benefits for repligen captiva primab resin with ligands modified with 5.2 kda and 21.5 kda peg chains, respectively. pegylation of proa ligands allowed the resin to retain a higher perc ...201829551237
development and validation of chemical features-based proton-coupled folate transporter/activity and reduced folate carrier/activity models (pharmacophores).all clinically used antifolates lack transport selectivity for tumors over normal cells resulting in dose-limiting toxicities. there is growing interest in developing novel tumor-targeted cytotoxic antifolates with selective transport into tumors over normal cells via the proton-coupled folate transporter (pcft) over the ubiquitously expressed reduced folate carrier (rfc). a lack of x-ray crystal structures or predictive models for pcft or rfc has hindered structure-aided drug design for pcft-se ...201829550744
process analytical technology for advanced process control in biologics manufacturing with the aid of macroscopic kinetic modeling.productivity improvements of mammalian cell culture in the production of recombinant proteins have been made by optimizing cell lines, media, and process operation. this led to enhanced titers and process robustness without increasing the cost of the upstream processing (usp); however, a downstream bottleneck remains. in terms of process control improvement, the process analytical technology (pat) initiative, initiated by the american food and drug administration (fda), aims to measure, analyze, ...201829547557
protein oxidative damage and redox imbalance induced by ionising radiation in cho cells.reactive oxygen species (ros) are important mediators of the cytotoxicity induced by the direct reaction of ionising radiation (ir) with all critical cellular components, such as proteins, lipids, and nucleic acids. the derived oxidative damage may propagate in exposed tissues in a dose- and spatiotemporal dependent manner to other cell compartments, affecting intracellular signalling, and cell fate. to understand how cell damage is induced, we studied the oxidative events occurring immediately ...201829546780
elucidating the impact of cho cell culture media on tryptophan oxidation of a monoclonal antibody through gene expression analyses.oxidation of monoclonal antibodies (mab) is a common chemical modification with potential impact on a therapeutic protein's activity and immunogenicity. in a previous study, it was found that tryptophan oxidation (trp-ox) levels of two mab produced in chinese hamster ovary (cho) cells were significantly lowered by modifying cell culture medium/feed. in this study, transcriptome analysis by rna-seq is applied to further elucidate the underlying mechanism of those changes in lowering the trp-ox le ...201829542860
evaluation of fluorescent dyes to measure protein aggregation within mammalian cell culture supernatants.a current challenge in bioprocessing is the ability to analyse critical quality attributes such as aggregation without prior purification. this study evaluated the use of fluorescent dyes (bis-ans, sypro orange, thioflavin t and proteostat) to characterise mab aggregates in chinese hamster ovary clarified cultures.201829540956
the concept of hybrid molecules of tacrine and benzyl quinolone carboxylic acid (bqca) as multifunctional agents for alzheimer's disease.novel tacrine-benzyl quinolone carboxylic acid (tacrine-bqca) hybrids were designed based on multi-target directed ligands (mtlds) paradigm, synthesized and evaluated in vitro as inhibitors of human acetylcholinesterase (hache) and human butyrylcholinesterase (hbche). tacrine moiety is represented herein as 7-methoxytacrine, 6-chlorotacrine or unsubstituted tacrine forming three different families of seven members, i.e. 21 compounds in overall. introducing bqca, a positive modulator of m1 muscar ...201829533874
the protective effect of niacinamide on cho aa8 cell line against ultraviolet radiation in the context of main cytoskeletal proteins.niacinamide is a stable and water-soluble form of vitamin b3, a valuable and versatile cosmetic ingredient, which is well absorbed and tolerated by the skin. a large body of literature has reported on the antioxidant and cell repair properties of niacinamide. therefore, it has been shown to be useful in the protection of the skin against ultraviolet b (uvb) radiation and free radicals. despite numerous hypotheses on the mechanism of vitamin b3, its protective effects have not yet been fully eluc ...201829533537
glycoengineering of mammalian expression systems on a cellular level.mammalian expression systems such as chinese hamster ovary (cho), mouse myeloma (ns0), and human embryonic kidney (hek) cells serve a critical role in the biotechnology industry as the production host of choice for recombinant protein therapeutics. most of the recombinant biologics are glycoproteins that contain complex oligosaccharide or glycan attachments representing a principal component of product quality. both n-glycans and o-glycans are present in these mammalian cells, but the engineerin ...201829532110
protective activity of pyruvate against vanadium-dependent cytotoxicity in chinese hamster ovary (cho-k1) cells.with increasing human exposure to vanadium-containing compounds and growing concern over their impact on human health, identification of safe methods for efficient treatment of vanadium poisoning may be of value. in this study, using chinese hamster ovary (cho-k1) cells, we show that the toxicity of vanadyl sulphate (voso4) is mitigated in the presence of sodium pyruvate. the exposure of cho-k1 cells to 100 μm voso4 for 48 h induced significant cytotoxicity (measured with a resazurin assay) and ...201829529943
oxidative stress and neurodegeneration: the possible contribution of quinone reductase 2.there is increasing evidence that oxidative stress is involved in the etiology and pathogenesis of neurodegenerative disorders. overproduction of reactive oxygen species (ros) is due in part to the reactivity of catecholamines, such as dopamine, adrenaline, and noradrenaline. these molecules are rapidly converted, chemically or enzymatically, into catechol-quinone and then into highly deleterious semiquinone radicals after 1-electron reduction in cells. notably, the overexpression of dihydronico ...201829526807
novel hghrh homodimer promotes fertility of female infertile hamster by up-regulating ovarian ghrh receptor without triggering gh secretion.extra-hypothalamic growth hormone-releasing hormone (ghrh) plays an important role in infertility. the female infertility models were formed by intraperitoneally injecting cyclophosphamide in 5-week-old chinese hamster once in a week for 5 weeks. all the models mated with healthy male hamster in the ratio of 1:1 in the experimental 6-8th week and the couples were separated to breed in the 9-10th week. 20 mg/kg of cyclophosphamide induced temporary interference of reproduction and did not cause s ...201829526766
protein a affinity chromatography of chinese hamster ovary (cho) cell culture broths containing biopharmaceutical monoclonal antibody (mab): experiments and mechanistic transport, binding and equilibrium modeling.protein a-based affinity chromatography is a highly-efficient separation method to capture, purify and isolate biosimilar monoclonal antibodies (mab) - an important medical product of biopharmaceutical industrial manufacturing. it is considered the most expensive step in purification downstream operations; therefore, its performance optimization offers a great cost saving in the overall production expenditure. the biochemical mixture-separating specific interaction experiments with chinese hamst ...201829522957
glyco-mapper: a chinese hamster ovary (cho) genome-specific glycosylation prediction tool.glyco-mapper is a novel systems biology product quality prediction tool created using a new framework termed: discretized reaction network modeling using fuzzy parameters (dream-zyp). within glyco-mapper, users fix the nutrient feed composition and the glycosylation reaction fluxes to fit the model glycoform to the reference experimental glycoform, enabling cell-line specific glycoform predictions as a result of cell engineering strategies. glyco-mapper accurately predicts glycoforms associated ...201829522825
a comparative study of using free radical generators in the testing of chosen oxidative stress parameters in the different types of cells.the aim of this study was to assess whether there are differences between the results of determining oxidative stress markers obtained from different origin cell lines after exposure to chemicals generating free radicals. the studies considered two markers of oxidative stress: the level of thiobarbituric acid reactive substances (tbars) and superoxide dismutase activity. the evaluation was performed in five cell lines: chinese hamster ovary (cho-9) cells, lung adenocarcinoma a549, macrophages ra ...201829519201
antibacterial effect of copaifera duckei dwyer oleoresin and its main diterpenes against oral pathogens and their cytotoxic effect.this study evaluates the antibacterial activity of the copaifera duckei dwyer oleoresin and two isolated compounds [eperu-8(20)-15,18-dioic acid and polyalthic acid] against bacteria involved in primary endodontic infections and dental caries and assesses the cytotoxic effect of these substances against a normal cell line. mic and mbc assays pointed out the most promising metabolites for further studies on bactericidal kinetics, antibiofilm activity, and synergistic antibacterial action. the ole ...201829515530
biogenesis of silver nanoparticles using leaf extract of indigofera hirsuta l. and their potential biomedical applications (3-in-1 system).the present study reports the biosynthesis of silver nanoparticles (ih-agnps) using aqueous leaf extract of indigofera hisruta l. the biosynthesized ih-agnps were found to be fcc crystals, 5-10 nm in size, spherical in shape and stable. the biosynthesized ih-agnps showed dose-dependant cytotoxicity against prostate cancer (pc3) (ic50 = 68.5 μg/ml), colon cancer (colo205) (ic50 = 85.2 μg/ml), and mouse melanoma (b16f10) (ic50 = 80.9 μg/ml). ih-agnps were found to be nontoxic towards normal cho (c ...201829513113
the combined effect of pdx1 overexpression and shh manipulation on the function of insulin-producing cells derived from adipose-tissue stem cells.pancreatic and duodenal homeobox 1 (pdx1) and sonic hedgehog (shh) are the key regulators of beta-cell function. in vitro experiments have shown that there is significant cooperation between pdx1 and shh with regard to the production and maintenance of insulin-producing cells (ipcs). in this study, the combined effect of pdx1 overexpression and shh manipulation on the function of adipose tissue-derived ipcs was determined. a eukaryotic expression vector (pdx1- pcdna3.1(+)) was constructed and tr ...201829511614
antioxidant activity of ge-132, a synthetic organic germanium, on cultured mammalian is a synthetic organic germanium that is used as a dietary supplement. the antioxidant activity of ge-132 on cultured mammalian cells was investigated in this study. first, ge-132 cytotoxicity on mammalian cultured cells was determined by measuring lactate dehydrogenase (ldh) levels. ge-132 had no cytotoxic effect on three different cell lines. second, the cell proliferative effect of ge-132 was determined by measuring atp content of whole cells and counting them. ge-132 treatment of chin ...201829503400
single copy transgene integration in a transcriptionally active site for recombinant protein synthesis.for the biomanufacturing of protein biologics, establishing stable cell lines with high transgene transcription is critical for high productivity. modern genome engineering tools can direct transgene insertion to a specified genomic locus and can potentially become a valuable tool for cell line generation. in this study, the authors survey transgene integration sites and their transcriptional activity to identify characteristics of desirable regions. a lentivirus containing destabilized green fl ...201830024101
toxicity and antigenotoxic effect of hispolon derivatives: role of structure in modulating cellular redox state and thioredoxin reductase.hispolon (hs), a bioactive polyphenol, and its derivatives such as hispolon monomethyl ether (hme), hispolon pyrazole (hp), and hispolon monomethyl ether pyrazole (hmep) were evaluated for comparative toxicity and antigenotoxic effects. the stability of hs derivatives in biological matrices followed the order hs < hp ≈ hme < hmep. the cytotoxicity analysis of hs derivatives indicated that hp and hmep were less toxic than hs and hme, respectively, in both normal and tumor cell types. the mechanis ...201830023935
car-t cells based on novel bcma monoclonal antibody block multiple myeloma cell growth.the cell-surface protein b cell maturation antigen (bcma, cd269) has emerged as a promising target for car-t cell therapy for multiple myeloma. in order to create a novel bcma car, we generated a new bcma monoclonal antibody, clone 4c8a. this antibody exhibited strong and selective binding to human bcma. bcma car-t cells containing the 4c8a scfv were readily detected with recombinant bcma protein by flow cytometry. the cells were cytolytic for rpmi8226, h929, and mm1s multiple myeloma cells and ...201830208593
chromawizard: an open source image analysis software for multicolor fluorescence in situ hybridization analysis.multicolor image analysis finds its applications in a broad range of biological studies. specifically, multiplex fluorescence in situ hybridization (m-fish) for chromosome painting facilitates the analysis of individual chromosomes in complex metaphase spreads and is widely used to detect both numerical and structural aberrations. while this is well established for human and mouse karyotypes, for which species sophisticated software and analysis tools are available, other organisms and species a ...201830089195
ph excursions impact cho cell culture performance and antibody n-linked excursions exist due to frequent base addition and environmental heterogeneity in large-scale bioreactors. such excursions could lead to suboptimal culture performance. here we investigated the impact of ph excursions on cell culture performance and n-linked glycosylation for three mab-producing chinese hamster ovary cell lines. frequent ph excursions were introduced by bolus base addition (in total 2-6% of initial volume, fixed bolus addition distributed from day 2 to 8) into small-scale bio ...201830088083
site specificity of eel luteinizing hormone n-linked oligosaccharides in signal transduction.eel luteinizing hormone (eellh) is composed of a common α-subunit and hormone specific β-subunit, both of which contain asparagine-linked carbohydrate residues, located at positions 56 and 79 on the α-subunit and position 10 on the β-subunit. the specific roles of the individual carbohydrate chains are poorly defined in eel. thus, we characterized the biologically active single chains by fusing the α-subunit to the carboxyl terminal region of the eellh β-subunit. site-directed mutagenesis of the ...201830056138
glyco-engineered cell line and computational docking studies reveals enterotoxigenic escherichia coli cfa/i fimbriae bind to lewis a glycans.we have previously reported clinical data to suggest that colonization factor i (cfa/i) fimbriae of enterotoxigenic escherichia coli (etec) can bind to lewis a (lea), a glycan epitope ubiquitous in the small intestinal mucosa of young children (<2 years of age), and individuals with a genetic mutation of fut2. to further elucidate the physiological binding properties of this interaction, we engineered chinese hamster ovary (cho-k1) cells to express lea or leb determinants on both n- and o-glycan ...201830050155
host cell protein profiling in biopharmaceutical harvests.biopharmaceuticals contain residual host cell protein (hcp) impurities, a complex mixture of endogenous proteins from production cell lines such as chinese hamster ovary (cho) cells. the composition of hcp impurities at harvest hinges on multiple factors, e.g., identity of cell line, cell density and viability at harvest, or other process parameters. two-dimensional differential gel electrophoresis (2-d dige) was used to compare hcp in 15 null cell culture harvest supernatants, which are represe ...201830048127
fluorescence "off" and "on" signalling of esculetin in the presence of copper and thiol: a possible implication in cellular thiol sensing.the interaction of the cupric ion with esculetin, a dihydroxy coumarin derivative, was studied by absorption and fluorescence spectroscopic methods in aqueous medium. esculetin formed a complex in the presence of the cupric ion which was characterised by the shift in its absorption band from 350 nm to 389 nm and the quenching of its fluorescence intensity at 466 nm. from job's plot and fluorescence quenching studies, the stoichiometry of the copper ion and esculetin in the complex was estimated ...201830043019
chlorotyrosines versus volatile byproducts from chlorine disinfection during washing of spinach and lettuce.following the food safety modernization act of 2011 in the u.s., guidelines for disinfection washes in food packaging facilities are under consideration to control pathogen risks. however, disinfectant exposures may need optimization because the high concentrations of chlorine disinfectant promote the formation of high levels of disinfection byproducts (dbps). when chlorine doses up through the 200 mg/l as cl2 range relevant to the current practice were applied to spinach and lettuce, significan ...201830040386
development of a shake tube-based scale-down model for perfusion cultures.the use of benchtop bioreactors (brs) for the development of mammalian cell perfusion cultures is expensive and time consuming, given its complexity in equipment and operation. scale-down models, going from liter to milliliter scale, are needed to support the rapid determination of suitable operating conditions in terms of viable cell density (vcd), perfusion rate, and medium composition. in this study, we compare the performance of steady-state perfusion cultures in orbitally shaken tube and br ...201830039852
culture temperature modulates half antibody and aggregate formation in a chinese hamster ovary cell line expressing a bispecific antibody.therapeutic bispecific antibodies are formed by assembly of multichain polypeptides. in general, a bispecific antibody has two different light chains and two different heavy chains that fold and correctly pair via engineered interchain interactions. because of some incorrect assembly, product-related impurities can be prevalent (e.g., half molecules, mispaired light chains, homodimers), requiring its removal during subsequent purification. in this study, we investigated the modulation of impurit ...201830036451
the contributions of individual galactosyltransferases to protein specific n-glycan processing in chinese hamster ovary cells.galactosylation as part of n-glycan processing is conducted by a set of beta-1,4-galactosyltransferases (b4galts), with b4galt1 as the dominant isoenzyme for this reaction. nevertheless, the exact contributions of this key-player as well as of the other isoenzymes involved in n-glycosylation, b4galt2, b4galt3 and b4galt4, have not been studied in-depth. to increase the understanding of the protein- and site-specific activities of individual galactosyltransferases in chinese hamster ovary cells, ...201830017654
genotoxicity and hemocompatibility of a novel calcium aluminate-based cement.the aim of this in vitro study was to evaluate the genotoxicity and hemocompatibility of a novel calcium aluminate-based cement, endobinder (eb) (binderware, são carlos, sp, brazil) and compare it with angelus white mineral trioxide aggregate (mta) (awmta) (angelus, soluções odontológicas, londrina, pr, brazil).201832161862
simulation and experimental study on the mechanism of the chlorination of azithromycin.azithromycin (azi) has been listed as an emerging contaminant by the us epa since 2009 because it is frequently detected in wastewater, surface water, and drinking water. in this paper, the chlorination of azi in drinking water was simulated and studied. the results indicated that new compounds were generated in the chlorination of azi. the byproducts were identified by liquid chromatography-quadrupole-time-of-flight mass spectrometry (lc-q-tof), and four of the byproducts were detected in real ...201830014912
generation of flp-intm-ready dg44 and lec cho cell lines for quick and easy constitutive protein expression.the well-characterized cell line chinese hamster ovary (cho) has been used to produce numerous biopharmaceuticals and is an important tool for basic research. however, introducing foreign dna into specially modified cho cells such as dg44 and lec can sometimes be an arduous process. here we show that the flp-intm plasmid can be modified to produce a fluorescent tracer protein tag (mcherrytm) as a fusion reporter, to allow for the rapid selection of single-cell sorted, isogenic flp-intm-r ...201830014730
differential expression of micrornas in recombinant chinese hamster ovary cells treated with sodium butyrate using digital rna counting.sodium butyrate (nabu) is an efficient supplement for increasing recombinant protein production in chinese hamster ovary (cho) cell culture. to elucidate the effects of nabu on mirna expression profile in recombinant cho (rcho) cells, differentially expressed mirnas in nabu-treated rcho cells were assessed by nanostring ncounter analysis. this result showed that eight mature mouse mirnas (let-7b, let-7d, mir-15b, mir-25, mir-27a, mir-99a, mir-125a-5p, and mir-125b-5p) were differentially express ...201830012463
improving the production of recombinant human bone morphogenetic protein-4 in chinese hamster ovary cell cultures by inhibition of undesirable endocytosis.endocytic regulation serves a critical role in modulating the extracellular level of signaling molecules, such as bone morphogenetic proteins (bmps). unfortunately, endocytosis may result in poor yields of recombinant human bmp-4 (rhbmp-4) from chinese hamster ovary (cho) cell cultures. when rhbmp-4 was incubated with cho cells, rhbmp-4 was actively internalized into cells. cell surface bound heparan sulfate proteoglycans (hspgs) served as the major receptors for rhbmp-4 internalization. removal ...201830011067
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