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in vitro antitrypanosomal and antiplasmodial activities of crude extracts and essential oils of ocimum gratissimum linn from benin and influence of vegetative stage.different parts of ocimum gratissimum linn are largely used in folk medicine for the treatment of many diseases, some of which related to parasitical infections as fevers and headaches. in order to validate their use and to clarify the plant part which possesses the best antiparasitic properties, we decided to evaluate the in vitro antiplasmodial and antitrypanosomal activities of essential oils and crude extracts from leaves, stems and seeds of ocimum gratissimum as well as their cytotoxicity.201425058875
a functional high-content mirna screen identifies mir-30 family to boost recombinant protein production in cho cells.the steady improvement of mammalian cell factories for the production of biopharmaceuticals is a key challenge for the biotechnology community. recently, small regulatory micrornas (mirnas) were identified as novel targets for optimizing chinese hamster ovary (cho) production cells as they do not add any translational burden to the cell while being capable of regulating entire physiological pathways. the aim of the present study was to elucidate mirna function in a recombinant cho-seap cell line ...201425061012
glycosylation-related genes in ns0 cells are insensitive to moderately elevated ammonium concentrations.ns0 and chinese hamster ovary (cho) cell lines are used to produce recombinant proteins for human therapeutics; however, ammonium accumulation can negatively impact cell growth, recombinant protein production, and protein glycosylation. to improve product quality and decrease costs, the relationship between ammonium and protein glycosylation needs to be elucidated. while ammonium has been shown to adversely affect glycosylation-related gene expression in cho cells, ns0 studies have not been perf ...201425062658
the fate of internalized α5 integrin is regulated by matrix-capable fibronectin.assembly of fibronectin matrices is associated with integrin receptor turnover and is an important determinant of tissue remodeling. although it is well established that fibronectin is the primary ligand for α5β1 receptor, the relationship between fibronectin matrix assembly and the fate of internalized α5 integrin remains poorly characterized.201425062814
wound healing potential of antibacterial microneedles loaded with green tea extracts.this study evaluates the utility of an antibacterial microneedle composed of green tea (gt) extract and hyaluronic acid (ha), for the efficient delivery of gt. these microneedles have the potential to be a patient-friendly method for the conventional sustained release of drugs. in this study, a fabrication method using a mold-based technique to produce gt/ha microneedles with a maximum area of ~50mm(2) with antibacterial properties was used to manufacture transdermal drug delivery systems. fouri ...201425063177
development of a soft-sensor based on multi-wavelength fluorescence spectroscopy and a dynamic metabolic model for monitoring mammalian cell cultures.a soft-sensor based on an extended kalman filter (ekf) that combines data obtained using a fluorescence-based soft-sensor with a dynamic mechanistic model, was investigated as a tool for continuous monitoring of a chinese hamster ovary (cho) cell cultivation process. a standalone fluorescence based soft-sensor, which uses a combination of an empirical multivariate statistical model and measured spectra, was designed for predicting key culture variables including viable and dead cells, recombinan ...201525065633
differential thermodynamic driving force of first- and second-generation antihistamines to determine their binding affinity for human h1 receptors.differential binding sites for first- and second-generation antihistamines were indicated on the basis of the crystal structure of human histamine h1 receptors. in this study, we evaluated differences between the thermodynamic driving forces of first- and second-generation antihistamines for human h1 receptors and their structural determinants. the binding enthalpy and entropy of 20 antihistamines were estimated with the van't hoff equation using their dissociation constants obtained from their ...201425065879
antioxidant potential of sutherlandia frutescens and its protective effects against oxidative stress in various cell cultures.sutherlandia frutescens (l.) r.br. (sf) is a south african plant that is widely used to treat stress, infections, cancer, and chronic diseases, many of which involve oxidative stress. the aim of the study was to quantitatively assess the antioxidant potential of sf extracts in cell-free system as well as in cell lines.201425070435
glyco-engineering for biopharmaceutical production in moss bioreactors.the production of recombinant biopharmaceuticals (pharmaceutical proteins) is a strongly growing area in the pharmaceutical industry. while most products to date are produced in mammalian cell cultures, namely chinese hamster ovary cells, plant-based production systems gained increasing acceptance over the last years. different plant systems have been established which are suitable for standardization and precise control of cultivation conditions, thus meeting the criteria for pharmaceutical pro ...201425071817
the transcription factor ap-1 regulates monkey 20α-hydroxysteroid dehydrogenase promoter activity in cho cells.monkey 20α-hydroxysteroid dehydrogenase (20α-hsd) is a catabolic enzyme responsible for converting progesterone into biologically inactive 20α-hydroxyprogesterone, thereby playing a key role in the estrous cycle or pregnancy and allowing ovulation and parturition to occur in most mammalian animals. monkey 20α-hsd was highly abundant in ovarian and placental tissues during the pre-ovulation and pre-parturition phase and was primarily localized in the syncytiotrophoblast of the placenta. in this s ...201425073972
optimal production of a fusion protein consisting of a single-chain variable fragment antibody against a tumor-associated antigen and interleukin-2 in fed-batch culture of pichia pastoris.the aim of the present study was to establish the strategy for producing a single-chain variable fragment (scfv) antibody fused with interleulin-2 (il2) by pichia pastoris and to optimize production during fed-batch cultivation in a 5-l fermenter.201425075014
high expression of fusion proteins consisting of a single-chain variable fragment antibody against a tumor-associated antigen and interleukin-2 in escherichia coli.the aim of this study was to establish a strategy for high-level production of single-chain variable fragment (scfv) antibodies fused with interleukin-2 (il-2) in escherichia coli.201425075015
characterization of the effects of cl⁻ channel modulators on tmem16a and bestrophin-1 ca²⁺ activated cl⁻ channels.the ca(2+) activated cl(-) channels (caccs) play a multitude of important physiological functions. a number of candidate proteins have been proposed to form cacc, but only two families, the bestrophins and the tmem16 proteins, recapitulate the properties of native cacc in expression systems. studies of endogenous caccs are hindered by the lack of specific pharmacology as most cl(-) channel modulators lack selectivity and a systematic comparison of the effects of these modulators on tmem16a and b ...201525078708
effects of glutamine and asparagine on recombinant antibody production using cho-gs cell lines.a unique and nontraditional approach using glutamine and asparagine supplements for cho-glutamine synthetase (gs) cell lines was studied. in our experiments, we found that a decrease in ph and an increase in cell death occurred in production phase of a gs cell line, leading to reduced antibody expression and lower antibody yields. the experimental results and the statistical analysis (anova) indicated that additions of glutamine and asparagine in the basal and feed media were effective to buffer ...201425079388
subcellular localization and function of mouse radial spoke protein 3 in mammalian cells and central nervous system.radial spoke protein 3 (rsp3) was first identified in chlamydomonas as a component of the radial spoke. the mammalian homologue of the chlamydomonas rsp3 gene is mainly expressed in testis and developing central nervous system (cns). however, the subcellular localization and function of mammalian rsp3 in the developing brain and mammalian cells remain poorly understood. here we show that the mouse rsp3 accumulates at the perinuclear region of chinese hamster ovary (cho) and 293t cells. detailed ...201425079589
discovery of transcription start sites in the chinese hamster genome by next-generation rna sequencing.chinese hamster ovary (cho) cell lines are one of the major production tools for monoclonal antibodies, recombinant proteins, and therapeutics. although many efforts have significantly improved the availability of sequence information for cho cells in the last years, forthcoming draft genomes still lack the information depth known from the mouse or human genomes. many genes annotated for cho cells and the chinese hamster reference genome still are in silico predictions, only insufficiently verif ...201425086342
combination of dextran sulfate and recombinant trypsin on aggregation of chinese hamster ovary cells.in laboratory scale therapeutical protein production, cell clumps form typically in shake flasks, which hinders cell growth and decreases protein yield. to minimize clumps during the culture of chinese hamster ovary cells, we employed the combination of two reagents, dextran sulfate (ds) and recombinant trypsin (r-trypsin). our results showed that both ds and r-trypsin could diminish cell aggregation when adding them respectively, but clumps were still noticed obviously. in order to further miti ...201625087075
the galnac-type o-glycoproteome of cho cells characterized by the simplecell strategy.the chinese hamster ovary cell (cho) is the major host cell factory for recombinant production of biological therapeutics primarily because of its "human-like" glycosylation features. cho is used for production of several o-glycoprotein therapeutics including erythropoietin, coagulation factors, and chimeric receptor igg1-fc-fusion proteins, however, some o-glycoproteins are not produced efficiently in cho. we have previously shown that the capacity for o-glycosylation of proteins can be one lim ...201425092905
bafilomycin l, a new inhibitor of cholesteryl ester synthesis in mammalian cells, produced by marine-derived streptomyces sp. opma00072.marine-derived streptomyces sp. opma00072 was found to produce inhibitors of the synthesis of neutral lipids in a cell-based assay using chinese hamster ovary (cho) cells. a new 16-membered macrolide named bafilomycin l (bfl) (1) was isolated along with the known structurally related bafilomycin c1 (bfc1) (3) from the culture broth of the actinomycete by solvent extraction, octadecylsilyl column chromatography and hplc. bfl inhibited cholesteryl ester (ce) synthesis in cho cells with an ic50 val ...201525095804
fluorescence-based soft sensor for at situ monitoring of chinese hamster ovary cell cultures.multi-wavelength fluorescence spectroscopy was investigated as a potential tool for use in monitoring key process variables that include: viable and dead cells, recombinant protein, glucose, and ammonia concentrations for chinese hamster ovary (cho) cells during cultivation.for the purpose of calibrating the fluorescence-based empirical model, cells were grown in batch mode with different initial glucose and glutamine concentrations.spectrofluorometer settings were optimized to ensure reproducib ...201425097916
it's time to regulate: coping with product-induced nongenetic clonal instability in cho cell lines via regulated protein expression.clonal instability and titer loss during chinese hamster ovary (cho) cell line development (cld) has several underlying causes, the most prominent of which are dna copy number loss and dna silencing. however, in some cases, clonal instability is due to the toxicity of the therapeutic protein(s) that clones express. unlike dna copy number loss, which may occur in some clones or dna silencing that is prevalent in certain regions of the genome, the hallmark of product induced clonal instability is ...201425104235
chronic atrial fibrillation increases microrna-21 in human atrial myocytes decreasing l-type calcium current.atrial fibrillation is characterized by progressive atrial structural and electrical changes (atrial remodeling) that favor arrhythmia recurrence and maintenance. reduction of l-type ca(2+) current (i(ca,l)) density is a hallmark of the electrical remodeling. alterations in atrial micrornas could contribute to the protein changes underlying atrial fibrillation-induced atrial electrical remodeling. this study was undertaken to compare mir-21 levels in isolated myocytes from atrial appendages obta ...201425107449
comparison of three different cell viability assays for evaluation of vanadyl sulphate cytotoxicity in a chinese hamster ovary k1 cell line.previously, evaluation of sodium metavanadate (navo3) cytotoxicity after 24 h exposure of chinese hamster ovary k1 (cho-k1) cells revealed different sensitivity of the in vitro assays used starting from the neutral red (nr, 3-amino-7-dimethylamino-2-methylphenazine hydrochloride) test (detecting lysosomal and possibly the golgi apparatus damage) as the most sensitive followed by the 2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2h-tetrazolium-5-carboxyanilide inner salt (xtt) and resazurin (7-hydroxy ...201625107459
vernakalant activates human cardiac k(2p)17.1 background k(+) channels.atrial fibrillation (af) contributes significantly to cardiovascular morbidity and mortality. the growing epidemic is associated with cardiac repolarization abnormalities and requires the development of more effective antiarrhythmic strategies. two-pore-domain k(+) channels stabilize the resting membrane potential and repolarize action potentials. recently discovered k2p17.1 channels are expressed in human atrium and represent potential targets for af therapy. however, cardiac electropharmacolog ...201425108155
mutagenicity and clastogenicity evaluation of metaphenoxy benzyl chloride by ames and micronucleus assays.metaphenoxy benzyl chloride (3-pbc; cas # 53874-66-1) is a commonly used intermediate in organic synthesis and is a widely used chemical building block. there is likelihood of human contact with 3-pbc in the process of organic or pesticide synthesis and even through as impurity in the product. further, this chemical has been reported to be found in water reservoirs causing a threat to aqueous flaura and fauna. genotoxic alert has been suspected for the chemical 3-pbc based on its structure hence ...201525109298
in vitro biological characterization of ifn-β-1a major glycoforms.recombinant human interferon β-1a (ifn-β-1a) is extensively used as the first-line treatment of relapsing forms of multiple sclerosis. its glycosylation is recognized as having a complex impact on a wide range of molecule characteristics and functions. the present study reports the enrichment of ifn-β-1a glycoforms and their physicochemical and biological characterization by means of electrospray ionization-mass spectrometry, sialic acid content, thermal denaturation and various in vitro bioassa ...201525117008
mammalian cell-produced therapeutic proteins: heterogeneity derived from protein degradation.therapeutic glycoproteins, for example, antibodies (abs) and fc fusion proteins when produced in mammalian cells, such as chinese hamster ovary (cho) cells generally exhibit heterogeneity. both the oligosaccharide moiety and the protein moiety contribute to this phenomenon. non-enzymatic and enzymatic pathways of protein fragmentation generate heterogeneity in the polypeptide backbone. in the non-enzymatic pathway, physical and chemical events such as light, oxidation, and others can cause the p ...201425118137
differential glycosylation of envelope gp120 is associated with differential recognition of hiv-1 by virus-specific antibodies and cell infection.hiv-1 entry into host cells is mediated by interactions between the virus envelope glycoprotein (gp120/gp41) and host-cell receptors. n-glycans represent approximately 50% of the molecular mass of gp120 and serve as potential antigenic determinants and/or as a shield against immune recognition. we previously reported that n-glycosylation of recombinant gp120 varied, depending on the producer cells, and the glycosylation variability affected gp120 recognition by serum antibodies from persons infe ...201425120578
xrcc2 rs3218536 polymorphism decreases the sensitivity of colorectal cancer cells to poly(adp-ribose) polymerase 1 inhibitor.single nucleotide polymorphisms (snps) are associated with the development of certain types of cancer. the present study aimed to investigate the association between x-ray repair complementing defective repair in chinese hamster cells 2 (xrcc2) snps and colorectal cancer (crc) cell sensitivity to the poly(adp-ribose) polymerase (parp) 1 inhibitor olaparib (azd2281). snapshot(®) analysis of xrcc2 snps was performed in five crc cell lines. the azd2281-sensitivities of the crc cells were also analy ...201425120693
high-throughput continuous flow femtosecond laser-assisted cell optoporation and transfection.we present a femtosecond-laser based nanoprocessing system for transient optical cell membrane poration to allow the introduction of foreign molecules into the interior of a cell with very high throughput. in the setup, cells flow through a micro-flow tube for spatial confinement and are simultaneously targeted by fs laser radiation. beam-shaping generates a focal geometry along a line which is scanned across the micro-flow cell to increase the number of reachable cells. successful cell membrane ...201425123087
reduction in c-terminal amidated species of recombinant monoclonal antibodies by genetic modification of cho cells.during development of recombinant monoclonal antibodies in chinese hamster ovary (cho) cells, c-terminal amidated species are observed. c-terminal amidation is catalysed by peptidylglycine α-amidating monooxygenase (pam), an enzyme known to be expressed in cho cells. the significant variations between clones during clone selection, and the relatively high content of amidated species (up to 15%) in comparison to reference material (4%), led us to develop a cell line with reduced production of c-t ...201425123359
effect of lithium chloride on the production and sialylation of fc-fusion protein in chinese hamster ovary cell culture.lithium chloride (licl), which is a specific inhibitor of glycogen synthase kinase-3β, is known to induce cell cycle arrest at the g2/m phase and to regulate apoptosis. to determine the potential of licl as a chemical additive to enhance specific productivity (q p) of recombinant chinese hamster ovary (rcho) cells through cell cycle arrest at g2/m phase, rcho cells producing fc-fusion protein were cultivated in serum-free media with licl concentrations ranging from 0 to 20 mm. the addition of li ...201425132065
cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics.protein glycosylation is an important post-translational modification toward the structure and function of recombinant therapeutics. the addition of oligosaccharides to recombinant proteins has been shown to greatly influence the overall physiochemical attributes of many proteins. it is for this reason that protein glycosylation is monitored by the developer of a recombinant protein therapeutic, and why protein glycosylation is typically considered a critical quality attribute. in this work, we ...201625132658
long-qt mutation p.k557e-kv7.1: dominant-negative suppression of iks, but preserved camp-dependent up-regulation.mutations in kcnq1, encoding for kv7.1, the α-subunit of the iks channel, cause long-qt syndrome type 1, potentially predisposing patients to ventricular tachyarrhythmias and sudden cardiac death, in particular, during elevated sympathetic tone. here, we aim at characterizing the p.lys557glu (k557e) kv7.1 mutation, identified in a dutch kindred, at baseline and during (mimicked) increased adrenergic tone.201425139741
residue 82 of the chikungunya virus e2 attachment protein modulates viral dissemination and arthritis in mice.chikungunya virus (chikv) is a mosquito-borne alphavirus that has reemerged to cause profound epidemics of fever, rash, and arthralgia throughout sub-saharan africa, southeast asia, and the caribbean. like other arthritogenic alphaviruses, mechanisms of chikv pathogenesis are not well defined. using the attenuated chikv strain 181/25 and virulent strain af15561, we identified a residue in the e2 viral attachment protein that is a critical determinant of viral replication in cultured cells and pa ...201425142598
design of serum-free medium for suspension culture of cho cells on the basis of general commercial media.the design of serum-free media for suspension culture of genetically engineered chinese hamster ovary (cho) cells using general commercial media as a basis was investigated. subcultivation using a commercial serum-free medium containing insulin-like growth factor (igf)-1 with or without fcs necessitated additives other than igf-1 to compensate for the lack of fcs and improve cell growth. suspension culture with media containing several combinations of growth factors suggested the effectiveness o ...201525149286
proteomics analysis of altered cellular metabolism induced by insufficient copper level.insufficient copper level in the mammalian cell culture medium resulted in lactate accumulation while maintaining similar growth and culture viability profiles. label-free, lc-ms/ms-based shotgun proteomics method was applied to compare the protein expression profiles obtained from the cultures exposed to suboptimal copper level to those provided with sufficient amount of copper. under copper deficient condition, a substantial reduction of the protein levels of the multiple subunits of complex i ...201425150618
direct real-time quantitative pcr for measurement of host-cell residual dna in therapeutic proteins.real-time quantitative pcr (qpcr) is important for quantification of residual host cell dna (resdna) in therapeutic protein preparations. typical qpcr protocols involve dna extraction steps complicating sample handling. here, we describe a "direct qpcr" approach without dna extraction. to avoid interferences of dna polymerase with a therapeutic protein, proteins in the samples were digested with proteinase k (pk) in the presence of sodium dodecyl sulfate (sds). tween 20 and nacl were included to ...201425151232
escherichia coli as a glycoprotein production host: recent developments and challenges.chinese hamster ovary cells are the most popular host expression system for the large-scale production of human therapeutic glycoproteins, but, the race to engineer escherichia coli to perform glycosylation is gathering pace. the successful functional transfer of an n-glycosylation pathway from campylobacter jejuni to escherichia coli in 2002 can be considered as the crucial first engineering step. here, we discuss the recent advancements in the field of n-glycosylation of recombinant therapeuti ...201425156401
methylene blue modulates β-secretase, reverses cerebral amyloidosis, and improves cognition in transgenic mice.amyloid precursor protein (app) proteolysis is required for production of amyloid-β (aβ) peptides that comprise β-amyloid plaques in the brains of patients with alzheimer disease (ad). here, we tested whether the experimental agent methylene blue (mb), used for treatment of methemoglobinemia, might improve ad-like pathology and behavioral deficits. we orally administered mb to the aged transgenic psapp mouse model of cerebral amyloidosis and evaluated cognitive function and cerebral amyloid path ...201425157105
accurate comparison of antibody expression levels by reproducible transgene targeting in engineered recombination-competent cho cells.over the years, chinese hamster ovary (cho) cells have emerged as the major host for expressing biotherapeutic proteins. traditional methods to generate high-producer cell lines rely on random integration(s) of the gene of interest but have thereby left the identification of bottlenecks as a challenging task. for comparison of different producer cell lines derived from various transfections, a system that provides control over transgene expression behavior is highly needed. this motivated us to ...201425158835
identification of antibody glycosylation structures that predict monoclonal antibody fc-effector function.to determine monoclonal antibody (mab) features that predict fragment crystalizable (fc)-mediated effector functions against hiv.201425160934
development and qualification of a high sensitivity, high throughput q-pcr assay for quantitation of residual host cell dna in purification process intermediate and drug substance samples.methods of high sensitivity, accuracy and throughput are needed for quantitation of low level residual host cell dna in purification process intermediates and drug substances of therapeutic proteins. in this study, we designed primer/probe sets targeting repetitive alu repeats or alu-equivalent sequences in the human, chinese hamster and murine genomes. when used in quantitative polymerase chain reactions (q-pcrs), these primer/probe sets showed high species specificity and gave significantly hi ...201425165010
diphenyl ditelluride-induced cell cycle arrest and apoptosis: a relation with topoisomerase i inhibition.the diphenyl ditelluride (dpdt) is a prototype for the development of new biologically active molecules. in previous studies, dpdt showed an elevated cytotoxicity in chinese hamster fibroblast (v79) cells but the mechanisms for reduction of cell viability still remain unknown. dpdt showed mutagenic properties by induction of frameshift mutations in bacterium salmonella typhimurium and yeast saccharomyces cerevisiae. this organotelluride also induced dna strand breaks in v79 cells. in this work, ...201525168415
use of slam and pvrl4 and identification of pro-hb-egf as cell entry receptors for wild type phocine distemper virus.signalling lymphocyte activation molecule (slam) has been identified as an immune cell receptor for the morbilliviruses, measles (mv), canine distemper (cdv), rinderpest and peste des petits ruminants (pprv) viruses, while cd46 is a receptor for vaccine strains of mv. more recently poliovirus like receptor 4 (pvrl4), also known as nectin 4, has been identified as a receptor for mv, cdv and pprv on the basolateral surface of polarised epithelial cells. pvrl4 is also up-regulated by mv in human br ...201425171206
biologically active constituents from salix viminalis bio-oil and their protective activity against hydrogen peroxide-induced oxidative stress in chinese hamster ovary cells.the protective antioxidative effect of the phenolic extract (pe) isolated from salix viminalis pyrolysis derived bio-oil was shown in vitro on the chinese hamster ovary (cho) cells exposed to hydrogen peroxide (h2o2). cells pretreated with 0.05 μg/ml pe after exposure to different concentrations of h2o2 (300-900 μm) showed up to 25 % higher viability than the unpretreated ones. the antioxidative effect of pe was also observed in a time-dependent manner. the results were confirmed by visual exami ...201425172057
in vitro evaluation of cordyceps militaris as a potential radioprotective agent.radiation is an important component of therapy for a wide range of malignant conditions. however, it triggers dna damage and cell death in normal cells and results in adverse side-effects. cordyceps militaris (c. militaris), a traditional medicinal mushroom, produces the bioactive compound, cordycepin (3'-deoxyadenosine) and has multiple pharmacological activities, such as antitumor, antimetastatic, antioxidant and immunomodulatory effects. the present study was undertaken to investigate whether ...201425176413
peroxisome biogenesis in mammalian cells.to investigate peroxisome assembly and human peroxisome biogenesis disorders (pbds) such as zellweger syndrome, thirteen different complementation groups (cgs) of chinese hamster ovary (cho) cell mutants defective in peroxisome biogenesis have been isolated and established as a model research system. successful gene-cloning studies by a forward genetic approach utilized a rapid functional complementation assay of cho cell mutants led to isolation of human peroxin (pex) genes. search for pathogen ...201425177298
protective effects of ascorbic acid against the genetic and epigenetic alterations induced by 3,5-dimethylaminophenol in aa8 cells.exposure to monocyclic aromatic alkylanilines (maas), namely 2,6-dimethylaniline (2,6-dma), 3,5-dimethylaniline (3,5-dma) and 3-ethylaniline (3-ea), was significantly and independently associated with bladder cancer incidence. 3,5-dmap (3,5-dimethylaminophenol), a metabolite of 3,5-dma, was shown to induce an imbalance in cytotoxicity cellular antioxidant/oxidant status, and dna damage in mammalian cell lines. this study was designed to evaluate the protective effect of ascorbic acid (asc) again ...201525178734
recombinant antibody color resulting from advanced glycation end product modifications.recombinant antibodies produced in chinese hamster ovary (cho) cells often exhibit a slight yellow-brown color, but the molecular basis for this color has remained elusive. we predicted that the color results from post-translational modifications on the antibody, because colored species were found to coelute with antibody products during size exclusion chromatography. previously, modification by oxidation and advanced glycation end products (ages) had been shown to give rise to colored and fluor ...201425181536
site-2 protease responds to oxidative stress and regulates oxidative injury in mammalian cells.site-2 protease (s2p) is a membrane-embedded protease that site-specifically cleaves intramembrane transcription factors, a necessary step for their maturation. s2p is well known to regulate cholesterol biosynthesis and endoplasmic reticulum stress in mammalian cells. in this study, we hypothesized that s2p could be responsible for the regulation of cellular oxidative injury under oxidative stress. wild type chinese hamster ovary (wt cho) cells and their mutant m19 cells with defective s2p gene ...201425183265
toxicity of drinking water disinfection byproducts: cell cycle alterations induced by the monohaloacetonitriles.haloacetonitriles (hans) are a chemical class of drinking water disinfection byproducts (dbps) that form from reactions between disinfectants and nitrogen-containing precursors, the latter more prevalent in water sources impacted by algae bloom and municipal wastewater effluent discharge. hans, previously demonstrated to be genotoxic, were investigated for their effects on the mammalian cell cycle. treating chinese hamster ovary (cho) cells with monohans followed by the release from the chemical ...201425185076
competitive mode and site of interaction of ticagrelor at the human platelet p2y12 -receptor.the g-protein-coupled p2y12 -receptor plays a crucial role in platelet aggregation. recently, ticagrelor was licensed as the first perorally active and reversible p2y12 -receptor antagonist.201425186974
meeting report-workshop on spike characterizations and virus removal by filtration: trends and new developments.this workshop was held on june 3, 2013, in berlin, germany, in conjunction with the pda virus & tse safety forum 2013. a total of nine speakers presented on key considerations of virus filtration, including a historical overview and emerging trends in evaluating parvovirus filters. several talks addressed understanding the mechanism of virus capture and breakthrough by filters, as well as addressing this risk by carefully controlling transmembrane pressure. improvements to validation studies wer ...201625188344
fabrication of swcnt-ag nanoparticle hybrid included self-assemblies for antibacterial applications.the present article reports the development of soft nanohybrids comprising of single walled carbon nanotube (swcnt) included silver nanoparticles (agnps) having superior antibacterial property. in this regard aqueous dispersing agent of carbon nanotube (cnt) containing a silver ion reducing unit was synthesised by the inclusion of tryptophan and tyrosine within the backbone of the amphiphile. the dispersions were characterized spectroscopically and microscopically using tem, afm and raman spectr ...201425191756
stabilization of cellular mitochondrial enzyme complex and sialyltransferase activity through supplementation of 30kc19 protein.in previous studies, 30kc19, a lipoprotein in silkworm hemolymph, enhanced productivity and glycosylation by expression of a 30kc19 gene or supplementation with a recombinant 30kc19 protein. additionally, 30kc19 exhibited enzyme-stabilizing and cell-penetrating abilities in vitro. in this study, we hypothesized that supplemented 30kc19 penetrated into the cell and enhanced the stability of the cellular enzyme. we investigated this using in vitro and cellular assessments. the activity of sialyltr ...201525193421
gene expression measurements normalized to cell number reveal large scale differences due to cell size changes, transcriptional amplification and transcriptional repression in cho cells.conventional approaches to differential gene expression comparisons assume equal cellular rna content among experimental conditions. we demonstrate that this assumption should not be universally applied because total rna yield from a set number of cells varies among experimental treatments of the same chinese hamster ovary (cho) cell line and among different cho cell lines expressing recombinant proteins. conventional normalization strategies mask these differences in cellular rna content and, c ...201425194670
[construction of controlled expression system of class b g-protein coupled receptor pac1].pac1 is the neuropeptide pituitary adenylate cyclase activating polypeptide (pacap) preferring receptor, which belongs to class b g protein-coupled receptors (gpcr) family. pac1 mediates the most effects of pacap as neurotransmitter, neuroregulator and neuroprotectant, while its high expression has close relationship with some physiological and pathological processes such as nerve-injury and tumor. to further understand the function of pac1, a cell line that expressed inducible pac1 was construc ...201425195253
structural basis of drugs that increase cardiac inward rectifier kir2.1 currents.we hypothesize that some drugs, besides flecainide, increase the inward rectifier current (ik1) generated by kir2.1 homotetramers (ikir2.1) and thus, exhibit pro- and/or antiarrhythmic effects particularly at the ventricular level. to test this hypothesis, we analysed the effects of propafenone, atenolol, dronedarone, and timolol on kir2.x channels.201425205296
detection of anti-aquaporin-4 autoantibodies in the sera of chinese neuromyelitis optica patients.in this study, we recruited 10 neuromyelitis optica patients, two multiple sclerosis patients and two myelitis patients. chinese hamster lung fibroblast (v79) cells transfected with a human aquaporin-4-mcherry fusion protein gene were used to detect anti-aquaporin-4 antibody in neuromyelitis optica patient sera by immunofluorescence. anti-aquaporin-4 autoantibody was stably detected by immunofluorescence in neuromyelitis optica patient sera exclusively. the sensitivity of the assay for neuromyel ...201325206717
[expression of neural cell adhesion molecule and modification of its n-glycan in ldld-14 cells].neural cell adhesion molecule (ncam) is a glycoprotein expressing on the surface of neurons, glial cells, bone cells and natural killer cells. ncam plays an important role in the process of cell - cell adhesion and cell migration, and is also a model protein to study polysialic acid. in this paper, ncam gene from mouse mammary gland cells (nmumg) was cloned into eukaryotic expression vectors pcdna3.1(+) and transfected into mutant chinese hamster ovary cells ldld-14. the stable transfection over ...201425212013
bodipy(®) fl histamine as a new modality for quantitative detection of histamine receptor upregulation upon ige sensitization in murine bone marrow-derived mast cells.flow cytometry is one of the most widely used methods for the qualitative and quantitative analysis of cell surface expressed proteins by making use of fluorescent specific antibodies. lacking an antibody validated for flow cytometry, an alternative approach for labeling cell surface receptors is the use of fluorescently tagged ligands. in this study, histamine h4 receptor transfected chinese hamster ovary cells and murine bone marrow-derived mast cells (mbmmcs) were selected for studying the po ...201525212523
effect of peptone feeding on transient gene expression process in cho dg44.transient gene expression (tge) gained popularity over the last decade as a rapid method for the production of milligram to gram quantities of recombinant proteins for preclinical studies in biophama industry. thereby, the optimization of the tge technique for chinese hamster ovary (cho) as the dominant host for the production of biotherapeutics is of great interest to reach the values for human embryo kidney-293 (hek-293) cells in terms of transfection efficiencies and production titers. tge ef ...201425215178
a robust transfection reagent for the transfection of cho and hek293 cells and production of recombinant proteins and lentiviral particles - ptg1.bioproduction of recombinant proteins (r-proteins) and recombinant lentiviral particles (r-lentiviral particles) requires robust transfections consisting of efficient protocols that are easy to implement, with good reproducibility for a maximum production of proteins and lentiviral particles in a short time with low cytotoxicity. this study evaluates the capacity of histidinylated polyethyleneimine i (ptg1) to facilitate robust dna transfection, with low cytotoxicity, of chinese hamster ovary (c ...201425215936
acute toxicity and genotoxicity study of fermented traditional herb formula guibi-tang.guibi-tang (guipi-tang in chinese and kihi-to in japanese) is a multi-herb traditional medicine commonly prescribed to treat psychoneurosis in east asia. although this medicine has been widely used, there is little available information on the safety and toxicity of guibi-tang, especially on the fermented one.201425218323
amino acid and glucose metabolism in fed-batch cho cell culture affects antibody production and glycosylation.fed-batch chinese hamster ovary (cho) cell culture is the most commonly used process for igg production in the biopharmaceutical industry. amino acid and glucose consumption, cell growth, metabolism, antibody titer, and n-glycosylation patterns are always the major concerns during upstream process optimization, especially media optimization. gaining knowledge on their interrelations could provide insight for obtaining higher immunoglobulin g (igg) titer and better controlling glycosylation-relat ...201525220616
dimerisation of the drosophila odorant coreceptor orco.odorant receptors (ors) detect volatile molecules and transform this external information into an intracellular signal. insect ors are heteromers composed of two seven transmembrane proteins, an odor-specific orx and a coreceptor (orco) protein. these ors form ligand gated cation channels that conduct also calcium. the sensitivity of the ors is regulated by intracellular signaling cascades. heterologously expressed orco proteins form also non-selective cation channels that cannot be activated by ...201425221476
functional bioassays for immune monitoring of high-risk neuroblastoma patients treated with ch14.18/cho anti-gd2 antibody.effective treatment of high-risk neuroblastoma (nb) remains a major challenge in pediatric oncology. human/mouse chimeric monoclonal anti-gd2 antibody (mab) ch14.18 is emerging as a treatment option to improve outcome. after establishing a production process in chinese hamster ovary (cho) cells, ch14.18/cho was made available in europe for clinical trials. here, we describe validated functional bioassays for the purpose of immune monitoring of these trials and demonstrate gd2-specific immune eff ...201425226154
raloxifene inhibits cloned kv4.3 channels in an estrogen receptor-independent manner.raloxifene is widely used for the treatment and prevention of postmenopausal osteoporosis. we examined the effects of raloxifene on the kv4.3 currents expressed in chinese hamster ovary (cho) cells using the whole-cell patch-clamp technique and on the long-term modulation of kv4.3 messenger rna (mrna) by real-time pcr analysis. raloxifene decreased the kv4.3 currents with an ic50 of 2.0 μm and accelerated the inactivation and activation kinetics in a concentration-dependent manner. the inhibitor ...201525231973
monitoring change in refractive index of cytosol of animal cells on affinity surface under osmotic stimulus for label-free measurement of viability.we demonstrated that a metal-clad waveguide (mcw)-based biosensor can be applied to label-free measurements of viability of adherent animal cells with osmotic stimulation in real time. after chinese hamster ovary (cho) and human embryonic kidney cell 293 (hek293) cells were attached to a concanavalin a (con a)-modified sensor surface, the magnitudes of cell responses to non-isotonic stimulation were compared between live and dead cells. the live cells exhibited a change in the refractive index ( ...201525232665
structure-activity profiles of novel 6-substituted pyrrolo[2,3-d]pyrimidine thienoyl antifolates with modified amino acids for cellular uptake by folate receptors α and β and the proton-coupled folate transporter.structure-activity relationships for cellular uptake and inhibition of cell proliferation were studied for 2-amino-4-oxo-6-substituted pyrrolo[2,3-d]pyrimidine thienoyl antifolates in which the terminal l-glutamate of the parent structure (7) was replaced by natural or unnatural amino acids. compounds 7 and 10-13 were selectively inhibitory toward folate receptor (fr) α-expressing chinese hamster ovary (cho) cells. antiproliferative effects of compounds 7 and 9-13 toward frα- and frβ-expressing ...201425234128
the tarantula toxin jingzhaotoxin-xi (κ-theraphotoxin-cj1a) regulates the activation and inactivation of the voltage-gated sodium channel nav1.5.specific peptide toxins interact with voltage-gated sodium channels by regulating the activation or inactivation of targeted channels. however, few toxins possessing dual effects have been identified. in the present study, we showed that jingzhaotoxin-xi/κ-theraphotoxin-cj1a (jztx-xi), a 34-residue peptide from the venom of the chinese spider chilobrachys jingzhao, inhibits the sodium conductance (ic50 = 124 ± 26 nm) and slows the fast inactivation (ec50 = 1.18 ± 0.2 μm) of nav1.5 expressed in c ...201425240294
mutations of rat surfactant protein a have distinct effects on its glycosylation, secretion, aggregation and degradation.surfactant protein a (sp-a) plays critical roles in the innate immune system and surfactant homeostasis of the lung. mutations in sp-a2 of the carbohydrate recognition domain (crd) impair its glycosylation and are associated with pulmonary fibrosis in humans. we aim to examine how mutations in sp-a that impair its glycosylation affect its biological properties and lead to disease.201425242514
genotoxicity of 1-methylpyrene and 1-hydroxymethylpyrene in chinese hamster v79-derived cells expressing both human cyp2e1 and sult1a1.1-methylpyrene (1-mp) is a widespread pollutant that is carcinogenic in animals following metabolic activation. previous studies have shown that benzylic hydroxylation of 1-mp, catalyzed by multiple cyp isoforms, gives rise to 1-hydroxymethylpyrene (1-hmp), which becomes bioreactive following further metabolism by various sulfotransferase (sult) isoforms. however, the mutagenic and chromosome damaging effects of 1-mp and 1-hmp in mammalian cells have not been investigated. in this study a chines ...201525243916
expression, purification, and characterization of recombinant mouse nerve growth factor in chinese hamster ovary cells.mouse ngf (mngf) extracted from mouse submaxillary gland has been approved on the market in china for treating nerve damage caused by n-hexane poisoning for over a decade, and many researches showed the clinical effectiveness of mngf for the treatment of other nerve system diseases. the extracted mngf have risks of potential viral contamination due to the animal origin. here, we report the successful expression, purification, and characterization of recombinant mngf (rmngf). an expression plasmi ...201425244938
characterization of membrane occupation and recognition nexus repeat containing 3, meiosis expressed gene 1 binding partner, in mouse male germ cells.mammalian spermatogenesis is a well-organized process of cell development and differentiation. meiosis expressed gene 1 (meig1) plays an essential role in the regulation of spermiogenesis. to explore potential mechanisms of meig1's action, a yeast two-hybrid screen was conducted, and several potential binding partners were identified; one of them was membrane occupation and recognition nexus repeat containing 3 (morn3). morn3 mrna is only abundant in mouse testis. in the testis, morn3 mrna is hi ...201725248657
phenylarsine oxide as a redox modulator of transient receptor potential vanilloid type 1 channel function.transient receptor potential vanilloid type 1 (trpv1) channels are capable of detecting and integrating noxious stimuli and play an important role in nociceptor activation and sensitization. it has been demonstrated that oxidizing agents are capable of positively modulating (sensitizing) the trpv1 channel. the present study investigates the ability of the thiol-oxidizing agent phenylarsine oxide (pao) to modulate trpv1 currents under voltage-clamp conditions. we assessed the ability of pao to mo ...201525250537
potentially lethal damage repair in drug arrested g2-phase cells after radiation exposure.potentially lethal damage (pld) repair has been defined as that property conferring the ability of cells to recover from dna damage depending on the postirradiation environment. using a novel cyclin dependent kinase 1 inhibitor ro-3306 to arrest cells in the g2 phase of the cell cycle, examined pld repair in g2 in cultured chinese hamster ovary (cho) cells. several cho-derived dna repair mutant cell lines were used in this study to elucidate the mechanism of dna double-strand break repair and to ...201425251700
spergularia marina induces glucagon-like peptide-1 secretion in nci-h716 cells through bile acid receptor activation.spergularia marina griseb. (sm) is a halophyte that grows in mud flats. the aerial portions of sm have been eaten as vegetables and traditionally used to prevent chronic diseases in korea. however, there has been no scientific report that demonstrates the pharmacological effects of sm. glucagon-like peptide-1 (glp-1) is important for the maintenance of glucose and energy homeostasis through acting as a signal in peripheral and neural systems. to discover a functional food for regulating glucose ...201425260089
function characterization of a glyco-engineered anti-egfr monoclonal antibody cetuximab in vitro.to evaluate the biochemical features and activities of a glyco-engineered form of the anti-human epidermal growth factor receptor monoclonal antibody (egfr mab) cetuximab in vitro.201425263334
modulation of hepatitis b surface antigen secretion by annexin ii expressed in hepatitis b virus‑producing hepatoma cells.the role of annexin ii in hepatitis b virus (hbv)‑associated hepatocellular carcinoma (hcc) remains to be elucidated. intracellular hepatitis b surface antigen (hbsag)‑retention contributes to the induction of hepatocarcinogenesis. the present study aimed to investigate the regulation of hbsag secretion by annexin ii expressed in hbv‑producing hepatoma cells. the expression of annexin ii was analyzed using western blot analysis in smmc‑7721, hepg2, hepg2.2.15, 293t and chinese hamster ovary (cho ...201425270731
hypoxia influences protein transport and epigenetic repression of cho cell cultures in shake flasks.shake flasks and bench-top bioreactors are widely used for cell culture process development, however, culture performances significantly differ between them. in order to apply the results received from small-scale cultures to production scale, it is important to understand the mechanisms underlying the differences between various culture systems. this study analyzes the expression patterns of chinese hamster ovary (cho) cells producing igg-fusion protein b0 cultured in shake flasks and 5-l bench ...201425271019
genomic organization and expression of immunoglobulin genes in the chinese hamster (cricetulus griseus).in science, the hamsters are widely used as a model for studying the human diseases because they display many features like humans. the utility of the chinese hamster as a biology model can be further enhanced by further characterization of the genes encoding components of the immune system. here, we report the genomic organization and expression of the chinese hamster immunoglobulin heavy and light chain genes. the chinese hamster igh locus contains 268 vh segments (132 potentially functional g ...201525271137
interaction of β-cyclodextrin with nile red in a single live cho cell: an initiative towards developing a prospective strategy for the excretion of adsorbed drugs from the cell membrane.a successful endeavour has been made to develop a prospective strategy to oust the drug molecules adsorbed on the cell membranes by simply using the non-toxic β-cyclodextrin (β-cd). for this purpose, fluorescent probes of different geometries and charge characteristics, namely phenosafranin (psf) and nile red (nr), were exploited for the in vitro studies using different lipids as model membranes. considering the success of the in vitro study, the present work is extended to a live chinese hamste ...201425274028
regulation of trpc6 channels by non-steroidal anti-inflammatory drugs.family focal segmental glomerulosclerosis (fsgs) is characterized by sclerosis and hyalinosis of particular loops of glomeruli and is one of the causes of the nephrotic syndrome. certain mutations in the structure of trpc6 channels are the genetic impetus for fsgs development resulting in podocytes functional abnormalities and various nephropathies. we have recently demonstrated that non-steroid anti-inflammatory drugs (nsaid) ibuprofen and diclofenac decrease the activity of endogenous trpc lik ...201225279100
enhanced cellular uptake of antisecretory peptide af-16 through proteoglycan binding.peptide af-16, which includes the active site of antisecretory factor protein, has antisecretory and anti-inflammatory properties, making it a potent drug candidate for treatment of secretory and inflammatory diseases such as diarrhea, inflammatory bowel diseases, and intracranial hypertension. despite remarkable physiological effects and great pharmaceutical need for drug discovery, very little is yet understood about af-16 mechanism of action. in order to address interaction mechanisms, we inv ...201425289567
peptides derived from central turn motifs within integrin αiib and αv cytoplasmic tails inhibit integrin activation.we previously found that peptides derived from the full length of integrin αiib and αv cytoplasmic tails inhibited their parent integrin activation, respectively. here we showed that the cell-permeable peptides corresponding to the conserved central turn motif within αiib and αv cytoplasmic tails, myr-krnrppleed (αiib peptide) and myr-krvrppqeeq (αv peptide), similarly inhibited both αiib and αv integrin activation. pre-treatment with αiib or αv peptides inhibited mn(2+)-activated αiibβ3 binding ...201425290158
backbone (1)h, (13)c, and (15)n resonance assignments of the fc fragment of human immunoglobulin g glycoprotein.the fc portion of immunoglobulin g (igg) recruits complements and its cognate receptors, thereby promoting defensive mechanisms in the humoral immune system. these effector functions critically depend on n-glycosylation at the fc region, which is therefore regarded as a crucial factor in the design and production of therapeutic antibodies. nmr spectroscopy plays a unique role in the characterization of conformational dynamics and intermolecular interactions of igg-fc in solutions. here, we repor ...201525291979
fluorescence-based, high-throughput assays for μ-opioid receptor activation using a membrane potential-sensitive dye.the development of new and improved opioid analgesics requires high-throughput screening (hts) methods to identify potential therapeutics from large libraries of lead compounds. here we describe two simple, real-time fluorescence-based assays of μ-opioid receptor activation that may be scaled up for hts. in att-20 cells expressing the μ-opioid receptor (mopr), opioids activate endogenous g protein gated inwardly rectifying k channels (girk channels), leading to membrane hyperpolarization. in chi ...201525293325
comparison of the biological effects of (18)f at different intracellular levels.we herein examined the biological effects of cells treated with (18)f labeled drugs for positron emission tomography (pet). the relationship between the intracellular distribution of (18)f and levels of damaged dna has yet to be clarified in detail. we used culture cells (chinese hamster ovary cells) treated with two types of (18)f labeled drugs, fluorodeoxyglucose (fdg) and fluorine ion (hf). fdg efficiently accumulated in cells, whereas hf did not. to examine the induction of dna double strand ...201425301551
safety assessment of epa-rich polar lipid oil produced from the microalgae nannochloropsis oculata.almega pl is an eicosapentaenoic acid-rich ω-3 oil that is isolated from nannochloropsis oculata algae and developed as a dietary supplement. the safety of the algal oil was evaluated in 14- and 90-day studies in sprague-dawley rats by oral gavage at dose levels of 0, 250, 500, and 2500 mg/kg/d and 0, 200, 400, and 2000 mg/kg/d, respectively. no mortalities occurred and no signs of toxicity were observed during the studies. no treatment-related effects were seen for body weight, food consumption ...201425305242
versatile cellular uptake mediated by catanionic vesicles: simultaneous spontaneous membrane fusion and endocytosis.lactose-derived catanionic vesicles offer unique opportunities to overcome cellular barriers. these potential nanovectors, very easy to formulate as drug delivery systems, are able to encapsulate drugs of various hydrophilicity. this article highlights versatile interaction mechanisms between these catanionic vesicles, labeled with hydrophilic and amphiphilic fluorescent probes, and a mammalian cell line, chinese hamster ovary. confocal microscopy and flow cytometry techniques show that these ve ...201525310849
peptone supplementation of culture medium has variable effects on the productivity of cho cells.the optimization of cell culture conditions for growth and productivity of recombinant chinese hamster ovary (cho) cells is a critical step in biopharmaceutical manufacturing. in the present study, the effects of the timing and amount of peptone feeding of a recombinant cho cell line grown in a basal medium in serum-free suspension culture were determined for eight peptones of different origin (plant and casein). the amino acid content and the average molecular weight of the peptones chosen were ...201425317401
naturally occurring mutations of human corticosteroid-binding globulin.corticosteroid-binding globulin (cbg) is encoded by serpina6. it is the major plasma binding protein of glucocorticoids and regulates plasma cortisol levels and bioavailability in humans. several proteases target cbg and disrupt its steroid-binding properties. to date, most genetic deficiencies that alter plasma cbg levels or function have been identified in patients presenting with a variety of clinical conditions.201525322275
on dynamically generating relevant elementary flux modes in a metabolic network using optimization.elementary flux modes (efms) are pathways through a metabolic reaction network that connect external substrates to products. using efms, a metabolic network can be transformed into its macroscopic counterpart, in which the internal metabolites have been eliminated and only external metabolites remain. in efms-based metabolic flux analysis (mfa) experimentally determined external fluxes are used to estimate the flux of each efm. it is in general prohibitive to enumerate all efms for complex netwo ...201525323319
salmeterol's extreme β2 selectivity is due to residues in both extracellular loops and transmembrane domains.salmeterol is a long-acting β2-agonist, widely used as an inhaled treatment of asthma and chronic obstructive pulmonary disease. it has very high β2-affinity (log kd -8.95) and is very selective for the β2-adrenoceptor (1000-fold selectivity over the β1-adrenoceptor). this study used a mutagenesis approach to determine the exact amino acids in the human β2-adrenoceptor responsible for this very high selectivity. wild-type β2- and β1-adrenoceptors, chimeric β2/β1-adrenoceptors, and receptors with ...201525324048
[expression, purification and functional identification of human psmp recombinant protein in chinese hamster ovary cells].to construct a new human chemotactic cytokine psmp eukaryotic expression vector to express psmp in chinese hamster ovary (cho) cells and to obtain the purified recombinant psmp protein for its functional mechanism study.201425331384
in vitro genotoxicity assessment of a novel resveratrol analogue, hs-1793.resveratrol has received considerable attention as a polyphenol with various biological effects such as anti-inflammatory, anti-oxidant, anti-mutagenic, anti-carcinogenic, and cardioprotective properties. as part of the overall safety assessment of hs-1793, a novel resveratrol analogue free from the restriction of metabolic instability and the high dose requirement of resveratrol, we assessed genotoxicity in three in vitro assays: a bacterial mutation assay, a comet assay, and a chromosomal aber ...201425343016
lqt1 mutations in kcnq1 c-terminus assembly domain suppress iks using different mechanisms.long qt syndrome 1 (lqt1) mutations in kcnq1 that decrease cardiac iks (slowly activating delayed rectifier k(+) current) underlie ventricular arrhythmias and sudden death. lqt1 mutations may suppress iks by preventing kcnq1 assembly, disrupting surface trafficking, or inhibiting gating. we investigated mechanisms underlying how three lqt1 mutations in kcnq1 c-terminus assembly domain (r555h/g589d/l619m) decrease iks in heterologous cells and cardiomyocytes.201425344363
differential complement activation pathways promote c3b deposition on native and acetylated ldl thereby inducing lipoprotein binding to the complement receptor 1.lipoproteins can induce complement activation resulting in opsonization and binding of these complexes to complement receptors. we investigated the binding of opsonized native ldl and acetylated ldl (acldl) to the complement receptor 1 (cr1). binding of complement factors c3b, igm, c1q, mannose-binding lectin (mbl), and properdin to ldl and acldl were investigated by elisa. subsequent binding of opsonized ldl and acldl to cr1 on cr1-transfected chinese hamster ovarian cells (cho-cr1) was tested ...201425349208
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