TitleAbstractYear(sorted ascending)
exploring the capabilities of fluorometric online monitoring on chinese hamster ovary cell cultivations producing a monoclonal monitoring of chinese hamster ovary fed-batch cell cultures via two-dimensional fluorescence spectroscopy (2dfs) was evaluated in this work. particular attention was directed toward different process strategies regarding the use of nutrient-rich feed media and temperature shifts. these intentionally performed process manipulations broadened the variances in the obtained fluorescence spectra and this was suspected to hamper the generation of reliable soft sensors. principal component analy ...201627390298
exposure to sirna-galnac conjugates in systems of the standard test battery for genotoxicity.registration of pharmaceuticals requires an assessment of their genotoxic potential using in vitro and in vivo tests outlined in the international conference on harmonisation (ich) guidance s2(r1). we have evaluated numerous sirna-n-acetylgalactosamine (galnac) conjugates containing phosphorothioate linkages and various combinations of 2'-fluoro and 2'-o-methyl ribose modifications of multiple nucleotides in the ich battery of assays, all of which have uniformly yielded negative results. to veri ...201627387336
heparin: past, present, and future.heparin, the most widely used anticoagulant drug in the world today, remains an animal-derived product with the attendant risks of adulteration and contamination. a contamination crisis in 2007-2008 increased the impetus to provide non-animal-derived sources of heparin, produced under cgmp conditions. in addition, recent studies suggest that heparin may have significant antineoplastic activity, separate and distinct from its anticoagulant activity, while other studies indicate a role for heparin ...201627384570
pingyangmycin and bleomycin share the same cytotoxicity pathway.pingyangmycin is an anticancer drug known as bleomycin a5 (a5), discovered in the pingyang county of zhejiang province of china. bleomycin (blm) is a mixture of mainly two compounds (a2 and b2), which is on the world health organization's list of essential medicines. both blm and a5 are hydrophilic molecules that depend on transporters or endocytosis receptors to get inside of cells. once inside, the anticancer activities rely on their abilities to produce dna breaks, thus leading to cell death. ...201627376254
augmenting chinese hamster genome assembly by identifying regions of high confidence.chinese hamster ovary (cho) cell lines are the dominant industrial workhorses for therapeutic recombinant protein production. the availability of genome sequence of chinese hamster and cho cells will spur further genome and rna sequencing of producing cell lines. however, the mammalian genomes assembled using shot-gun sequencing data still contain regions of uncertain quality due to assembly errors. identifying high confidence regions in the assembled genome will facilitate its use for cell engi ...201627374913
percutaneous treatment of herniated lumbar discs with ozone: investigation of the mechanisms of elucidate the mechanism of action of intradiscal oxygen-ozone therapy for herniated intervertebral disc therapy.201627363296
[domestication of suspension cho cells and its application in the expression of anti-psma antibody].to domesticate adherent chinese hamster ovary (cho) cells into suspension cho cells (cho-s) cultured in serum-free medium,and evaluate the application of the cho-s cells in antibody expression.201627358992
the infectious bac genomic dna expression library: a high capacity vector system for functional genomics.gene dosage plays a critical role in a range of cellular phenotypes, yet most cellular expression systems use heterologous cdna-based vectors which express proteins well above physiological levels. in contrast, genomic dna expression vectors generate physiologically-relevant levels of gene expression by carrying the whole genomic dna locus of a gene including its regulatory elements. here we describe the first genomic dna expression library generated using the high-capacity herpes simplex virus- ...201627353647
subchronic and genetic safety evaluation of a calcium fructoborate in rats.a branded calcium fructoborate product, a nature-identical calcium salt of bis (fructose) ester of boric acid found in plants and a natural source of boron in the human diet and sold under the trade name fruitex-b(®) brand calcium fructoborate ("frxb"), was evaluated in a 90-day dietary toxicity study and two genotoxicity studies. in the 90-day study, four groups of 10 male and 10 female crl:sd cd(®) igs rats were fed diets with frxb admixtures of 0.56, 1.12, and 1.68% dietary concentration, pro ...201627350145
effects of heterocyclic-based head group modifications on the structure-activity relationship of tocopherol-based lipids for non-viral gene delivery.gene therapy, a promising strategy for the delivery of therapeutic nucleic acids, is greatly dependent on the development of efficient vectors. in this study, we designed and synthesized several tocopherol-based lipids varying in the head group region. here, we present the structure-activity relationship of stable aqueous suspensions of lipids that were synthetically prepared and formulated with 1,2-dioleoyl phosphatidyl ethanolamine (dope) as the co-lipid. the physicochemical properties such as ...201627348545
a theoretical estimate for nucleotide sugar demand towards chinese hamster ovary cellular glycosylation.glycosylation greatly influences the safety and efficacy of many of the highest-selling recombinant therapeutic proteins (rtps). in order to define optimal cell culture feeding strategies that control rtp glycosylation, it is necessary to know how nucleotide sugars (nss) are consumed towards host cell and rtp glycosylation. here, we present a theoretical framework that integrates the reported glycoproteome of cho cells, the number of n-linked and o-galnac glycosylation sites on individual host c ...201627345611
genotoxicity biomarkers for airborne particulate matter (pm2.5) in an area under petrochemical influence.the effects of fine inhalable particles (pm2.5) were evaluated in an area under the influence of a petrochemical industry, investigating the sensitivity of different genotoxicity biomarkers. organic extracts were obtained from pm2.5 samples at two sites, positioned in the first and second preferential wind direction in the area. the extracts were evaluated with salmonella/microsome assay, microsuspension method, strains ta98, yg1021 and yg1024. the mammalian metabolization fraction (s9) was used ...201627343868
understanding and controlling sialylation in a cho fc-fusion process.a chinese hamster ovary (cho) bioprocess, where the product is a sialylated fc-fusion protein, was operated at pilot and manufacturing scale and significant variation of sialylation level was observed. in order to more tightly control glycosylation profiles, we sought to identify the cause of variability. untargeted metabolomics and transcriptomics methods were applied to select samples from the large scale runs. lower sialylation was correlated with elevated mannose levels, a shift in glucose m ...201627310468
hyperosmotic stimulus study discloses benefits in atp supply and reveals mirna/mrna targets to improve recombinant protein production of cho cells.biopharmaceuticals are predominantly produced by chinese hamster ovary (cho) cells cultivated in fed-batch mode. hyperosmotic culture conditions (≥ 350 mosmol kg(∑1) ) resulting from feeding of nutrients may enhance specific product formation rates (qp ). as an improved atp supply was anticipated to enhance qp this study focused on the identification of suitable mirna/mrna targets to increase atp levels. therefor next generation sequencing and a compartment specific metabolomics approach were ap ...201627214792
quantitative definition and monitoring of the host cell protein proteome using itraq - a study of an industrial mab producing cho-s cell line.there are few studies defining cho host cell proteins (hcps) and the flux of these throughout a downstream purification process. here we have applied quantitative itraq proteomics to follow the hcp profile of an antibody (mab) producing cho-s cell line throughout a standard downstream purification procedure consisting of a protein a, cation and anion exchange process. we used both 6 sample itraq experiment to analyze technical replicates of three samples, which were culture harvest (hccf), prote ...201627214759
real-time monitoring of specific oxygen uptake rates of embryonic stem cells in a microfluidic cell culture device.oxygen plays a key role in stem cell biology as a signaling molecule and as an indicator of cell energy metabolism. quantification of cellular oxygen kinetics, i.e. the determination of specific oxygen uptake rates (sours), is routinely used to understand metabolic shifts. however current methods to determine sour in adherent cell cultures rely on cell sampling, which impacts on cellular phenotype. we present real-time monitoring of cell growth from phase contrast microscopy images, and of respi ...201627214658
discovery and characterization of antibody variants using mass spectrometry-based comparative analysis for biosimilar candidates of monoclonal antibody drugs.liquid chromatography mass spectrometry (lc-ms) is the most commonly used technique for the characterization of antibody variants. mab-x and mab-y are two approved igg1 subtype monoclonal antibody drugs recombinantly produced in chinese hamster ovary (cho) cells. we report here that two unexpected and rare antibody variants have been discovered during cell culture process development of biosimilars for these two approved drugs through intact mass analysis. we then used comprehensive mass spectro ...201627214604
defective association of the platelet glycoprotein ib-ix complex with the glycosphingolipid-enriched membrane domain inhibits murine thrombus and atheroma formation.localization of the platelet glycoprotein ib-ix complex to the membrane lipid domain is essential for platelet adhesion to von willebrand factor and subsequent platelet activation in vitro. yet, the in vivo importance of this localization has never been addressed. we recently found that the disulfide linkage between ibα and ibβ is critical for the association of ibα with the glycosphingolipid-enriched membrane domain; in this study, we established a transgenic mouse model expressing this mutant ...201627206768
mutagenic properties of linuron and chlorbromuron evaluated by means of cytogenetic biomarkers in mammalian cell lines.agricultural practices are usually supported by several chemical substances, such as herbicides. linuron and chlorbromuron are phenylurea herbicides largely used to protect crops from weeds, blocking photosynthesis by inhibition of the photosystem ii complex. the former, also commercially known as lorox or afalon, is selectively used to protect bean and french bean plants, fennels, and celeriacs; the second, commercially known as maloran, is selectively used for carrots, peas, potatoes, soy spro ...201627206752
comprehensive characterization of reference standard lots of hiv-1 subtype c gp120 proteins for clinical trials in southern african regions.two hiv-1 subtype c gp120 protein candidates were the selected antigens for several experimental vaccine regimens now under evaluation in hvtn 100 phase i/ii clinical trial aiming to support the start of the hvtn 702 phase iib/iii trial in southern africa, which is designed to confirm and extend the partial protection seen against hiv-1 infection in the rv144 thai trial. here, we report the comprehensive physicochemical characterization of the gp120 reference materials that are representative of ...201627187483
in vitro evaluation of 2-hydroxyalkylated β-cyclodextrins as potential therapeutic agents for niemann-pick type c disease.this study was conducted to evaluate the attenuating potential of 2-hydroxypropyl-β-cyclodextrin (hpbcd) against niemann-pick type c (npc) disease, as well as the physical and chemical properties, particularly the cholesterol-solubilizing ability, in an npc disease model in vitro. as parameters of npc abnormalities, intracellular free and esterified cholesterol levels and lysosome volume were measured in npc1 null chinese hamster ovary cells. hpbcd showed dose-dependent effects against dysfuncti ...201627184436
disruption of the gene c12orf35 leads to increased productivities in recombinant cho cell lines.recently, we reported that the loss of a telomeric region of chromosome 8 in chinese hamster ovary (cho) cells correlates with higher recombinant productivities. new cell lines lacking this region, called cho-c8del, showed several advantages during cell line generation and for the production of recombinant proteins (ritter et al., 2016, biotechnol bioeng). here, we performed knock-down and knock-out experiments of genes located within this telomeric region of chromosome 8 to identify the genes c ...201627183150
exposure to 50hz-sinusoidal electromagnetic field induces dna damage-independent electromagnetic field (emf) is commonly encountered within our daily lives, the biological effects of emf are of great concern. autophagy is a key process for maintaining cellular homeostasis, and it can also reveal cellular responses to environmental stimuli. in this study, we aim to investigate the biological effects of a 50hz-sinusoidal electromagnetic field on autophagy and we identified its mechanism of action in chinese hamster lung (chl) cells. chl cells were exposed to a 50hz sinusoid ...201627177844
organelle-targeted h2s probes enable visualization of the subcellular distribution of h2s donors.hydrogen sulfide (h2s) is an essential biological signaling molecule in diverse biological regulatory pathways. to provide new chemical tools for h2s imaging, we report here a fluorescent h2s detection platform (hsn2-bg) that is compatible with subcellular localization snap-tag fusion protein methodologies and use appropriate fusion protein constructs to demonstrate mitochondrial and lysosomal localization. we also demonstrate the efficacy of this detection platform to image endogenous h2s in ch ...201627171507
accelerated homology-directed targeted integration of transgenes in chinese hamster ovary cells via crispr/cas9 and fluorescent enrichment.targeted gene integration into site-specific loci can be achieved in chinese hamster ovary (cho) cells via crispr/cas9 genome editing technology and the homology-directed repair (hdr) pathway. the low efficiency of hdr often requires antibiotic selection, which limits targeted integration of multiple genes at multiple sites. to improve hdr-mediated targeted integration, while avoiding the use of selection markers, chemical treatment for increased hdr, and fluorescent enrichment of genome-edited ...201627159230
apolipoprotein e lipoprotein particles inhibit amyloid-β uptake through cell surface heparan sulphate proteoglycan.the accumulation, aggregation and deposition of amyloid-β (aβ) peptides in the brain are central to the pathogenesis of alzheimer's disease (ad). alzheimer's disease risk increases significantly in individuals carrying one or two copies of apoe ε4 allele compared to individuals with an ε3/ε3 genotype. growing evidence has demonstrated that apolipoprotein e (apoe) strongly influences ad pathogenesis by controlling aβ aggregation and metabolism. heparan sulphate proteoglycans (hspgs) are abundant ...201627151330
highly luminescent heterostructured copper-doped zinc sulfide nanocrystals for application in cancer cell labeling.the structural characteristics of the seed-mediated synthesis of heterostructured cus-zns nanocrystals (ncs) and cu-doped zns (zns:cu) ncs synthesized by two different protocols are compared and analyzed. at high cu dopant concentrations, segregated subclusters of zns and cus are observed. the photoluminescence quantum yield of zns:cu ncs is about 50-80 %; a value much higher than that of zns ncs (6 %). finally, these ncs are coated with a thin silica shell by using (3-mercaptopropyl)triethoxysi ...201627146419
classification of normal and apoptotic cells from fluorescence microscopy images using generalized polynomial chaos and level set function.accurate automated quantitative analysis of living cells based on fluorescence microscopy images can be very useful for fast evaluation of experimental outcomes and cell culture protocols. in this work, an algorithm is developed for fast differentiation of normal and apoptotic viable chinese hamster ovary (cho) cells. for effective segmentation of cell images, a stochastic segmentation algorithm is developed by combining a generalized polynomial chaos expansion with a level set function-based se ...201627142234
combined sodium ion sensitivity in agonist binding and internalization of vasopressin v1b receptors.reducing na(+) in the extracellular environment may lead to two beneficial effects for increasing agonist binding to cell surface g-protein coupled receptors (gpcrs): reduction of na(+)-mediated binding block and reduce of receptor internalization. however, such combined effects have not been explored. we used chinese hamster ovary cells expressing vasopressin v1b receptors as a model to explore na(+) sensitivity in agonist binding and receptor internalization. under basal conditions, a large fr ...201627138239
asymmetrical flow field-flow fractionation coupled with multi-angle laser light scattering for stability comparison of virus-like particles in different solution environments.the stabilities of two commercially available virus like particles, cho-hbsag expressed by chinese hamster ovary (cho) cells and hans-hbsag expressed by hansenula polymorpha (hans), were compared using af4-malls under different treatment processes. the initial molecular weight and hydrodynamic diameter of cho-hbsag measured with af4-malls were 4727kda and 29.4nm, while those of hans-hbsag were 3039kda and 22.8nm respectively. in salt solution of 2m ammonium sulfate, the molecular weight and size ...201627129428
[stable expression of human anti-il-33 scfv-igg1fc fusion protein in cho k1 cells].to construct two different eukaryotic expression vectors of human anti-interleukin 33 (il-33) single-chain antibody fragment (scfv-fc) to transfect chinese hamster ovary (cho) k1 cells and select the stably and high-level expressed cell lines to improve the expression level of the fusion protein.201627126936
neuro-immune interactions in chemical-induced airway hyperreactivity.asthma may be induced by chemical sensitisers, via mechanisms that are still poorly understood. this type of asthma is characterised by airway hyperreactivity (ahr) and little airway inflammation. since potent chemical sensitisers, such as toluene-2,4-diisocyanate (tdi), are also sensory irritants, it is suggested that chemical-induced asthma relies on neuro-immune mechanisms.we investigated the involvement of transient receptor potential channels (trp) a1 and v1, major chemosensors in the airwa ...201627126687
enhanced recombinant factor vii expression in chinese hamster ovary cells by optimizing signal peptides and fed-batch medium.signal peptides play an important role in directing and efficiently transporting secretory proteins to their proper locations in the endoplasmic reticulum of mammalian cells. the aim of this study was to enhance the expression of recombinant coagulation factor vii (rfvii) in cho cells by optimizing the signal peptides and type of fed-batch culture medium used. five sub-clones (o2, i3, h3, g2 and m3) with different signal peptide were selected by western blot (wb) analysis and used for suspension ...201627116572
expression and bioactivity of recombinant human serum albumin and dtmp fusion proteins in cho cells.the 14-amino acid (iegptlrqwlaara) thrombopoietin mimetic peptide (tmp) shares no sequence homology with native thrombopoietin (tpo). when dimerized, it displays a high-binding affinity for the tpo receptor and has equipotent bioactivity with recombinant human tpo (rhtpo) in stimulating proliferation and maturation of megakaryocytes in vitro. however, tmp is limited for clinical usage because of its short half-life in vivo. in this study, fusion proteins that composed of tandem dimer of tmp (dtm ...201627115755
study on physiological roles of stimulation of prostaglandin e2 receptor subtype ep2 in urethral function in rats.we investigated the relaxant effect of stimulation of prostaglandin e2 (pge2 ) receptor subtype ep2 as well as the involvement of a cyclic amp (camp)-dependent pathway related to stimulation of ep2 receptors in urethral function in rats by evaluating effects of pge2 and selective ep2 receptor agonist cp-533,536.201627111625
identifying the differences in mechanisms of mycophenolic acid controlling fucose content of glycoproteins expressed in different cho cell the biopharmaceutical industry, glycosylation is a critical quality attribute that can modulate the efficacy of a therapeutic glycoprotein. obtaining a consistent glycoform profile is desired because molecular function can be defined by its carbohydrate structures. specifically, the fucose content of oligosaccharides in glycoproteins is one of the most important attributes that can significantly affect antibody-dependent cellular cytotoxicity (adcc) activity. it is therefore important to unde ...201627093551
cb1 cannabinoid receptor-mediated increases in cyclic amp accumulation are correlated with reduced gi/o function.cb1 cannabinoid receptors (cb1rs) stimulate gi/o-dependent signaling pathways. cb1r-mediated camp increases were proposed to result from gs activation, but cb1r-stimulated gtpγs binding to gs has not heretofore been investigated.201627089415
mutation detection in an antibody-producing chinese hamster ovary cell line by targeted rna sequencing.chinese hamster ovary (cho) cells have been used widely in the pharmaceutical industry for production of biological therapeutics including monoclonal antibodies (mab). the integrity of the gene of interest and the accuracy of the relay of genetic information impact product quality and patient safety. here we employed next-generation sequencing, particularly rna-seq, and developed a method to systematically analyze the mutation rate of the mrna of cho cell lines producing a mab. the effect of an ...201627088091
dimerization of melanocortin receptor 1 (mc1r) and mc5r creates a ligand-dependent signal modulation: potential participation in physiological color change in the flounder.vertebrates produce α-melanocyte-stimulating hormone (α-msh), which contains an n-terminal acetyl group, and desacetyl-α-msh, which does not contain an n-terminal acetyl group. in teleosts and amphibians, α-msh-related peptides stimulate pigment dispersion via melanocortin receptors 1-5 (mc1r-mc5r), which are members of the g-protein-coupled receptor (gpcr) family. we previously reported an interesting phenomenon associated with physiological color changes in the skin of a flatfish, barfin floun ...201627080548
identification of drug-like inhibitors of insulin-regulated aminopeptidase through small-molecule screening.intracerebroventricular injection of angiotensin iv, a ligand of insulin-regulated aminopeptidase (irap), has been shown to improve cognitive functions in several animal models. consequently, irap is considered a potential target for treatment of cognitive disorders. to identify nonpeptidic irap inhibitors, we adapted an established enzymatic assay based on membrane preparations from chinese hamster ovary cells and a synthetic peptide-like substrate for high-throughput screening purposes. the 38 ...201627078680
the long intron 1 of growth hormone gene from reeves' turtle (chinemys reevesii) correlates with negatively regulated gh expression in four cell lines.turtles grow slowly and have a long lifespan. ultrastructural studies of the pituitary gland in reeves' turtle (chinemys reevesii) have revealed that the species possesses a higher nucleoplasmic ratio and fewer secretory granules in growth hormone (gh) cells than other animal species in summer and winter. c. reevesii gh gene was cloned and species-specific similarities and differences were investigated. the full gh gene sequence in c. reevesii contains 8517 base pairs (bp), comprising five exons ...201627077853
one-step conjugation method for site-specific antibody-drug conjugates through reactive cysteine-engineered antibodies.engineered cysteine residues are particularly convenient for site-specific conjugation of antibody-drug conjugates (adc), because no cell engineering and additives are required. usually, unpaired cysteine residues form mixed disulfides during fermentation in chinese hamster ovarian (cho) cells; therefore, additional reduction and oxidization steps are required prior to conjugation. in this study, we prepared light chain (lc)-q124c variants in igg and examined the conjugation efficiency. intrigui ...201627074832
comprehensive genome and epigenome characterization of cho cells in response to evolutionary pressures and over time.the most striking characteristic of cho cells is their adaptability, which enables efficient production of proteins as well as growth under a variety of culture conditions, but also results in genomic and phenotypic instability. to investigate the relative contribution of genomic and epigenetic modifications towards phenotype evolution, comprehensive genome and epigenome data are presented for six related cho cell lines, both in response to perturbations (different culture conditions and media a ...201627072894
quantitative analysis of the supernatant from host and transfected cho cells using itraq 8-plex technique.we employed uplc-ms/ms with itraq 8-plex labeling to quantitatively analyze the supernatant produced by two chinese hamster ovary (cho) cell lines (cho k1sv and cho cat-s). in each case, the supernatant from the host and three transfected clones were analyzed at days 5, 7, and 10 of culture. a total of eight itraq 8-plex experiments were performed. for each cell line, the overlap of supernatant protein identifications between transfected clones is over 60%. over 70% of the supernatant proteins i ...201627070921
isotetrandrine reduces astrocyte cytotoxicity in neuromyelitis optica by blocking the binding of nmo-igg to aquaporin 4.neuromyelitis optica (nmo) is a severe neurological demyelinating autoimmune disease that affects the optic nerves and spinal cord with no cure and no fda-approved therapy. research over the last decade revealed that the binding of nmo-igg to the water channel protein astrocyte aquaporin 4 (aqp4) might be the primary cause of nmo pathogenesis. the purpose of this study was to identify potential blockers of nmo-igg and aqp4 binding.201627064690
characterization of recombinant human diamine oxidase (rhdao) produced in chinese hamster ovary (cho) cells.human diamine oxidase (hdao) efficiently degrades polyamines and histamine. reduced enzyme activities might cause complications during pregnancy and be involved in histamine intolerance. so far hdao has been characterized after isolation from either native sources or the heterologous production in insect cells. accessibility to human enzyme is limited and insect cells produce non-human glycosylation patterns that may alter its biochemical properties. we present the heterologous expression of hda ...201627063138
poly-pathway model, a novel approach to simulate multiple metabolic states by reaction network-based model - application to amino acid depletion in cho cell culture.mammalian cell lines are characterized by a complex and flexible metabolism. a single model that could describe the variations in metabolic behavior triggered by variations in the culture conditions would be a precious tool in bioprocess development. in this paper, we introduce an approach to generate a poly-pathway model and use it to simulate diverse metabolic states triggered in response to removal, reduction or doubling of amino acids in the culture medium of an antibody-producing cho cell l ...201627060554
polysorbates 20 and 80 degradation by group xv lysosomal phospholipase a2 isomer x1 in monoclonal antibody formulations.decreases in polysorbate (ps80) content were observed while evaluating the long-term storage stability of chinese hamster ovary-derived, purified monoclonal antibodies. it was determined that polysorbate had been enzymatically degraded; therefore, studies were performed to identify and characterize the protein(s) responsible. polysorbate degrading activity was enriched from chinese hamster ovary media leading to the identification of group xv lysosomal phospholipase a2 isomer x1 (lpla2) by shotg ...201627056628
development of betulinic acid as an agonist of tgr5 receptor using a new in vitro assay.g-protein-coupled bile acid receptor 1, also known as tgr5 is known to be involved in glucose homeostasis. in animal models, treatment with a tgr5 agonist induces incretin secretion to reduce hyperglycemia. betulinic acid, a triterpenoid present in the leaves of white birch, has been introduced as a selective tgr5 agonist. however, direct activation of tgr5 by betulinic acid has not yet been reported.201627578964
a new family of insect muscarinic acetylcholine receptors.most currently used insecticides are neurotoxic chemicals that target a limited number of sites and insect cholinergic neurotransmission is the major target. a potential target for insecticide development is the muscarinic acetylcholine receptor (machr), which is a metabotropic g-protein-coupled receptor. insects have a- and b-type machrs and the five mammalian machrs are close to the a-type. we isolated a cdna (cg12796) from the fruit fly, drosophila melanogaster. after heterologous expression ...201627003873
mitochondrial genome of cricetulus migratorius (rodentia: cricetidae): insights into the characteristics of the mitochondrial genome and the phylogenetic relationships of cricetulus species.cricetulus migratorius is widely distributed in the northwestern arid regions of china. here, the complete mitochondrial genome (mitogenome) of c. migratorius is reported, to our knowledge, for the first time. it was found to be 16,246bp in length, including 13 protein-coding genes (pcgs), 2 rrna genes, 22 trna genes, and one control region, and showed characteristics typical of the vertebrate mitogenome. comparative analyses of mitogenomes of cricetulus species showed that hamster mitogenomes h ...201627717890
structural and antitrypanosomal data of different carbasones of piperitone.this article reports data on four carbazones of piperitone: semicarbazone 1, thiosemicarbazone 2, 4-phenyl semicarbazone 3 and 4-phenyl thiosemicarbazone 4 prepared directly in situ from essential oil of cymbopogon schoenantus, whose gc-fid and gc-ms analysis revealed piperitone as major component (68.20%). the structures of hemi-synthesized compounds were confirmed by high throughput ir, ms, (1)h and (13)c nmr based spectrometric analysis. their antiparasitic activities were evaluated in vitro ...201627900358
phasor plotting with frequency-domain flow cytometry.interest in time resolved flow cytometry is growing. in this paper, we collect time-resolved flow cytometry data and use it to create polar plots showing distributions that are a function of measured fluorescence decay rates from individual fluorescently-labeled cells and fluorescent microspheres. phasor, or polar, graphics are commonly used in fluorescence lifetime imaging microscopy (flim). in flim measurements, the plotted points on a phasor graph represent the phase-shift and demodulation of ...201627410612
[anti-hbs persistence after revaccination with three doses of hepatitis b vaccine among non-responsive adults: a 4-year of follow-up study].to explore anti-hbs persistence four years after revaccination with hepatitis b vaccine (hepb) among adults who were non-responsive to hepb primary immunization.201627256728
[anti-hbs persistence following revaccination with three doses of hepatitis b vaccine among low-responsive adults after primary vaccination: a 4-year follow-up study].to assess the 4-year anti-hbs persistence after revaccination with 3-dose of hepatitis b vaccine (hepb) among low-responsive adults.201627256727
[antibody persistence following primary vaccination with hepatitis b vaccine among normal and high-responder adults: a 5-year follow-up study].to evaluate the 5-year antibody persistence and the risk factors associated with the persistence after primary vaccination of hepatitis b vaccine (hepb) among normal or high-response adults.201627256726
[anti-hbs persistence following primary vaccination with three doses of hepatitis b vaccine among normal and high-responder adults: a 3-year follow-up study].to assess the 3-year anti-hbs persistence after primary vaccination with three-dose of hepatitis b vaccine (hepb) among normal and high-responder adults.201627256725
impact of hydrolysates on monoclonal antibody productivity, purification and quality in chinese hamster ovary cells.plant and yeast derived hydrolysates are economical and efficient alternative medium supplements to improve mammalian cell culture performance. we supplemented two commercial chinese hamster ovary (cho) culture media with hydrolysates from four different sources, yeast, soybean, ex-cell cd (a chemically defined hydrolysate replacement) and wheat to improve the productivity of two cell lines expressing different monoclonal antibodies (mabs). yeast, soybean and ex-cell cd improved the final mab ti ...201627067279
newcastle disease virus hemagglutinin neuraminidase as a potential cancer targeting agent.the hemagglutinin-neuraminidase (hn) protein of newcastle disease virus (ndv) with its immunotherapeutic activities and sialic acid binding abilities is a promising cancer adjuvant. the hn was surfaced displayed on lactococcus lactis and its cancer targeting ability was investigated via attachment to the mda-mb231 breast cancers. to surface display the hn protein on the bacterial cell wall, hn was fused to n-acetylmuraminidase (acma) anchoring motif of l. lactis and expressed in chinese hamster ...201626918060
design, synthesis, and in vitro antituberculosis activity of 2(5h)-furanone derivatives.a series of 2(5h)-furanone-based compounds were synthesized from commercially available mucohalic acids. from the first-generation compounds, three showed inhibitory activity (10 µg/ml) of at least 35% against mycobacterium smegmatis mc(2) 155 growth (bioscreen c system). in screening the active first-generation compounds for growth inhibition against mycobacterium tuberculosis h37rv, the most active compound was identified with a minimum inhibitory concentration (mic99 ) of 8.07 µg/ml (15.8 µm) ...201627346745
construction of a novel dna vaccine candidate encoding an hspx-ppe44-esxv fusion antigen of mycobacterium tuberculosis.mycobacterium tuberculosis is the causative agent of tuberculosis (tb). bacille calmette-guerin (bcg) vaccine, is not effective in adults, therefore, many efforts have been made to produce an effective adult tb vaccine. the aim of this study was to develop a new tuberculosis dna vaccine candidate encoding a recombinant hspx-ppe44-esxv fusion antigen of m. tuberculosis.201627536702
highly efficient transfer of chromosomes to a broad range of target cells using chinese hamster ovary cells expressing murine leukemia virus-derived envelope proteins.microcell-mediated chromosome transfer (mmct) is an essential step for introducing chromosomes from donor cells to recipient cells. mmct allows not only for genetic/epigenetic analysis of specific chromosomes, but also for utilization of human and mouse artificial chromosomes (hacs/macs) as gene delivery vectors. although the scientific demand for genome scale analyses is increasing, the poor transfer efficiency of the current method has hampered the application of chromosome engineering technol ...201627271046
removal process of prion and parvovirus from human platelet lysates used as clinical-grade supplement for ex vivo cell expansion.pooled human platelet lysate (hpl) is becoming the new gold standard as supplement for ex vivo cell culture for clinical protocols. however, the risk of pathogen contamination of hpl increases with the platelet pool size. we hypothesized that hollow fiber anion exchange membrane chromatography using qyuspeed d (qsd) could remove resistant and untested bloodborne pathogens, such as parvoviruses and prions, from hpl-supplemented growth media without substantially affecting their capacity to suppor ...201627260210
synthesis and biological evaluation of a series of non-hemiacetal ester derivatives of an attempt to improve the efficacy and stability of current, clinically used artemisinins, a series non-hemiacetal ester derivatives of artemisinin were synthesized and evaluated for their in vitro antiplasmodial and anticancer activities as well as cytotoxicities. these esters were synthesized through the reaction of acid anhydrides, or acid chlorides with artemisinin derived alcohol. in vitro antiplasmodial activity assessments were conducted against intraerythrocytic nf54 and dd2 plasmodiu ...201627448920
generation and characterization of a bivalent hiv-1 subtype c gp120 protein boost for proof-of-concept hiv vaccine efficacy trials in southern africa.the viral envelope glycoprotein (env) is the major target for antibody (ab)-mediated vaccine development against the human immunodeficiency virus type 1 (hiv-1). although several recombinant env antigens have been evaluated in clinical trials, only the surface glycoprotein, gp120, (from hiv-1 subtype b, mn, and subtype crf_01ae, a244) used in the alvac prime-aidsvax gp120 boost rv144 phase iii hiv vaccine trial was shown to contribute to protective efficacy, although modest and short-lived. henc ...201627442017
xrcc1 rs25487 genetic variant and tp53 mutation at codon 249 predict clinical outcomes of hepatitis b virus-related hepatocellular carcinoma after hepatectomy: a cohort study for 10 years' follow investigate the effects of rs25487 (the dna repair gene: x-ray repair complementing defective repair in chinese hamster cells 1 [xrcc1]) and codon 249 mutation (tp53 gene) on clinical outcomes of post-hepatectomy hepatitis b virus (hbv)-related hcc.201626519348
influenza virus hemagglutinin glycoproteins with different n-glycan patterns activate dendritic cells in vitro.influenza virus hemagglutinin (ha) n-glycans play important regulatory roles in the control of virus virulence, antigenicity, receptor-binding specificity, and viral escape from the immune response. considered essential for controlling innate and adaptive immune responses against influenza virus infections, dendritic cells (dcs) trigger proinflammatory and adaptive immune responses in hosts. in this study, we engineered chinese hamster ovary (cho) cell lines expressing recombinant ha from pandem ...201627099319
liposome-mediated herpes simplex virus uptake is glycoprotein-d receptor-independent but requires heparan sulfate.cationic liposomes are widely used to facilitate introduction of genetic material into target cells during transfection. this study describes a non-receptor mediated herpes simplex virus type-1 (hsv-1) entry into the chinese hamster ovary (cho-k1) cells that naturally lack glycoprotein d (gd)-receptors using a commercially available cationic liposome: lipofectamine. presence of cell surface heparan sulfate (hs) increased the levels of viral entry indicating a potential role of hs in this mode of ...201627446014
high levels of histone h3 acetylation at the cmv promoter are predictive of stable expression in chinese hamster ovary cells.chinese hamster ovary cells (cho) are widely used in the production of glycosylated therapeutic proteins such as antibodies. during expansion and maintenance, cho cell lines are prone to production instability, which may be caused by promoter silencing, loss of transgene copies, or post-transcriptional effects. silencing of recombinant genes may be accompanied by dna methylation and histone modification. previously, we demonstrated that cytosine methylation of the human cytomegalovirus major imm ...201627073121
novel epstein-barr virus-like particles incorporating gh/gl-ebna1 or gb-lmp2 induce high neutralizing antibody titers and ebv-specific t-cell responses in immunized mice.previous epstein-barr virus (ebv) prophylactic vaccines based on the major surface glycoprotein gp350/220 as an immunogen have failed to block viral infection in humans, suggesting a need to target other viral envelope glycoproteins. in this study, we reasoned that incorporating gh/gl or gb, critical glycoproteins for viral fusion and entry, on the surface of a virus-like particle (vlp) would be more immunogenic than gp350/220 for generating effective neutralizing antibodies to prevent viral inf ...201627926486
rabbits immunized with epstein-barr virus gh/gl or gb recombinant proteins elicit higher serum virus neutralizing activity than gp350.epstein-barr virus (ebv) is the primary cause of infectious mononucleosis and has been strongly implicated in the etiology of multiple epithelial and lymphoid cancers, such as nasopharyngeal carcinoma, gastric carcinoma, hodgkin lymphoma, burkitt lymphoma, non-hodgkin lymphoma and post-transplant lymphoproliferative disorder. there is currently no licensed prophylactic vaccine for ebv. most efforts to develop prophylactic vaccines have focused on ebv gp350, which binds to cd21/cd35 to gain entry ...201627291087
monocyclic β-lactam and unexpected oxazinone formation: synthesis, crystal structure, docking studies and antibacterial evaluation.novel monocyclic β-lactam derivatives bearing aryl, phenyl and heterocyclic rings were synthesized as possible antibacterial agents. cyclization of imines (3h, 3t) with phenylacetic acid in the presence of phosphoryl chloride and triethyl amine did not afford the expected β-lactams. instead, highly substituted 1,3-oxazin-4-ones (4h, 4t) were isolated as the only product and confirmed by single crystal x-ray analysis of 4t. the results of antibacterial activity showed that compound 4l exhibited c ...201626133357
recombinant production and characterization of human anti-influenza virus monoclonal antibodies identified from hybridomas fused with human previous studies, hybridomas producing human immunoglobulin g, the antibodies 5e4 and 5a7 against influenza a and b virus were established using a novel human lymphocyte fusion partner, spymeg. in the present study, we succeeded in achieving the recombinant production and secretion of 5e4 and 5a7 in chinese hamster ovary cells. our n-glycan analysis by intact-mass detection and liquid chromatography mass spectrometry showed that recombinant 5e4 and 5a7 have one n-glycan and the typical mammal ...201627464991
endocytic function is critical for influenza a virus infection via dc-sign and l-sign.the ubiquitous presence of cell-surface sialic acid (sia) has complicated efforts to identify specific transmembrane glycoproteins that function as bone fide entry receptors for influenza a virus (iav) infection. the c-type lectin receptors (clrs) dc-sign (cd209) and l-sign (cd209l) enhance iav infection however it is not known if they act as attachment factors, passing virions to other unknown receptors for virus entry, or as authentic entry receptors for clr-mediated virus uptake and infection ...201626763587
dc-sign and l-sign are attachment factors that promote infection of target cells by human metapneumovirus in the presence or absence of cellular is well established that glycosaminoglycans (gags) function as attachment factors for human metapneumovirus (hmpv), concentrating virions at the cell surface to promote interaction with other receptors for virus entry and infection. there is increasing evidence to suggest that multiple receptors may exhibit the capacity to promote infectious entry of hmpv into host cells; however, definitive identification of specific transmembrane receptors for hmpv attachment and entry is complicated by the ...201627334579
detection and quantification of adp-ribosylated rhoa/b by monoclonal antibody.clostridium botulinum exoenzyme c3 is the prototype of c3-like adp-ribosyltransferases that modify the gtpases rhoa, b, and c. c3 catalyzes the transfer of an adp-ribose moiety from the co-substrate nicotinamide adenine dinucleotide (nad) to asparagine-41 of rho-gtpases. although c3 does not possess cell-binding/-translocation domains, c3 is able to efficiently enter intact cells, including neuronal and macrophage-like cells. conventionally, the detection of c3 uptake into cells is carried out v ...201627043630
biophysical studies on calcium and carbohydrate binding to carbohydrate recognition domain of gal/galnac lectin from entamoeba histolytica: insights into host cell adhesion.entamoeba histolytica, an enteric parasite expresses a gal/galnac-specific lectin that contributes to its virulence by establishing adhesion to host cell. in this study, carbohydrate recognition domain of hgl (ehcrd) was purified and biophysical studies were conducted to understand the thermodynamic basis of its binding to carbohydrate and ca(++) here, we show that carbohydrate recognition domain (crd) of the lectin binds to calcium through dpn motif. to decipher the role of calcium in carbohydr ...201627008865
rapid screening by cell-based fusion assay for identifying novel antivirals of glycoprotein b-mediated herpes simplex virus type 1 infection.herpes simplex virus type 1 (hsv-1) is a causative agent for a variety of diseases. although antiherpetic drugs such as acyclovir have been developed to inhibit virus replication through interaction with dna kinases, their continuous administration leads to an increase in the frequency of drug-resistant hsv-1, which is an important clinical issue that requires urgent solution. recently, we reported that the sialylated o-linked sugar t antigen (stn) and its attached peptide region (o-glycosylated ...201627803463
bioisosteric ferrocenyl-containing quinolines with antiplasmodial and antitrichomonal properties.bioisosteric ferrocenyl-containing quinolines and ferrocenylamines containing organosilanes and their carbon analogues, were prepared and fully characterised. the molecular structures of two ferrocenyl-containing quinolines, determined using single-crystal x-ray diffraction, revealed that the compounds crystallise in a folded conformation. the compounds were screened for their antiplasmodial activity against the chloroquine-sensitive (nf54) and cq-resistant (dd2) strains of p. falciparum, as wel ...201627858010
general and genetic toxicology of enzyme-treated ginseng extract: toxicology of ginseng rh2.ginseng rh2+ is enzyme-treated ginseng extract containing high amounts of converted ginsenosides, such as compound k, rh2, rg3, which have potent anticancer activity. we conducted general and genetic toxicity tests to evaluate the safety of ginseng rh2+.201627695630
fast filtration of bacterial or mammalian suspension cell cultures for optimal metabolomics results.the metabolome offers real time detection of the adaptive, multi-parametric response of the organisms to environmental changes, pathophysiological stimuli or genetic modifications and thus rationalizes the optimization of cell cultures in bioprocessing. in bioprocessing the measurement of physiological intracellular metabolite levels is imperative for successful applications. however, a sampling method applicable to all cell types with little to no validation effort which simultaneously offers h ...201627438065
calibrated complex impedance of cho cells and e. coli bacteria at ghz frequencies using scanning microwave microscopy.the application of scanning microwave microscopy (smm) to extract calibrated electrical properties of cells and bacteria in air is presented. from the s 11 images, after calibration, complex impedance and admittance images of chinese hamster ovary cells and e. coli bacteria deposited on a silicon substrate have been obtained. the broadband capabilities of smm have been used to characterize the bio-samples between 2 ghz and 20 ghz. the resulting calibrated cell and bacteria admittance at 19 ghz w ...201626895571
multi-omics data driven analysis establishes reference codon biases for synthetic gene design in microbial and mammalian this study, we analyzed multi-omics data and subsets thereof to establish reference codon usage biases for codon optimization in synthetic gene design. specifically, publicly available genomic, transcriptomic, proteomic and translatomic data for microbial and mammalian expression hosts, escherichia coli, saccharomyces cerevisiae, pichia pastoris and chinese hamster ovary (cho) cells, were compiled to derive their individual codon and codon pair frequencies. then, host dependent and -omics spe ...201626850284
[study of in vitro expression of human platelet itgb3 gene nonsense mutation c.1476g>a].to explore the function of a novel nonsense mutation c.1476g>a of itgb3 gene using an in vitro expression system.201626829726
web application for genetic modification flux with database to estimate metabolic fluxes of genetic mutants.computational analysis of metabolic fluxes is essential in understanding the structure and function of a metabolic network and in rationally designing genetically modified mutants for an engineering purpose. we had presented the genetic modification flux (gmf) that predicts the flux distribution of a broad range of genetically modified mutants. to enhance the feasibility and usability of gmf, we have developed a web application with a metabolic network database to predict a flux distribution of ...201626777238
two-mab cocktail protects macaques against the makona variant of ebola virus.the 2014-2015 ebola virus (ebov) outbreak in west africa highlighted the urgent need for specific therapeutic interventions for infected patients. the human-mouse chimeric monoclonal antibody (mab) cocktail zmapp, previously shown to be efficacious in ebov (variant kikwit) lethally infected nonhuman primates (nhps) when administration was initiated up to 5 days, was used in some patients during the outbreak. we show that a two-antibody cocktail, mil77e, is fully protective in nhps when administe ...201626962157
cho cell production and sequence improvement in the 13c6fr1 anti-ebola antibody.from march 2014 through february 2015, the ebola virus spread rapidly in west africa, resulting in almost 30,000 infections and approximately 10,000 deaths. with no approved therapeutic options available, an experimental antibody cocktail known as zmapp™ was administered to patients on a limited compassionate-use basis. the supply of zmapp™ was highly constrained at the time because it was in preclinical development and a novel production system (tobacco plants) was being used for manufacturing. ...201626761424
high-efficiency secretory expression of human neutrophil gelatinase-associated lipocalin from mammalian cell lines with human serum albumin signal peptide.human neutrophil gelatinase associated lipocalin (ngal) is a secretory glycoprotein initially isolated from neutrophils. it is thought to be involved in the incidence and development of immunological diseases and cancers. urinary and serum levels of ngal have been investigated as a new biomarker of acute kidney injury (aki), for an earlier and more accurate detection method than with creatinine level. however, expressing high-quality recombinant ngal is difficult both in escherichia coli and mam ...201626518367
ultrastructural changes in a-cells exposed to diabetic hyperglycaemia. observations made on pancreas of chinese hamsters. 20164914660
effect of circadian rhythm on the radiation response of the chinese hamster (cricetulus griseus). 20165030088
n-ethylmaleimide and the cyclic response to x-rays of synchronous chinese hamster cells. 20164736799
effects of isoleucine deficiency on nucleic acid and protein metabolism in cultured chinese hamster cells. continued ribonucleic acid and protein synthesis in the absence of deoxyribonucleic acid synthesis. 20165061874
studies on induced aberrations in diplochromosomes of chinese hamster cells. 20164796813
the retinal vasculature of the chinese hamster: a preliminary study. 20164914659
intermittent dna synthesis and periodic expression of enzyme activity in the cell cycle of wi-38.synchronous cultures of wi-38 were obtained using an automated system for detachment and partitioning of mitotic cells which operates without the use of inhibitors, altered medium, or lowered temperatures. the generation time in synchronous wi-38 is 19.5 h and the duration of s phase when determined from the percentage of labeled metaphase cells or nuclei is 12 h. dna replication in wi-38 occurs in three temporally distinct and rapid bursts separated by intervals of greatly reduced synthesis wit ...20164795861
antioxidant and dna repair stimulating effect of extracts from transformed and normal roots of rhaponticum carthamoides against induced oxidative stress and dna damage in cho cells.rhaponticum carthamoides has a long tradition of use in siberian folk medicine. the roots and rhizomes of this species are used in various dietary supplements or nutraceutical preparations to increase energy level or eliminate physical weakness. this is the first report to reveal the protective and dna repair stimulating abilities of r. carthamoides root extracts in chinese hamster ovary (cho) cells exposed to an oxidative agent. both transformed root extract (tr extract) and extract of soil-gro ...201627034736
[anatomical studies on the urinary tract and genitalia of the chinese dwarf hamster (cricetulus griseus)]. 20165036384
islet and beta cell volumes in offspring of severely diabetic (ketotic) chinese hamsters. 20164914657
temporal variation of adenine ribonucleotides during the cell cycle of chinese hamster fibroblasts in culture. 20164855877
chromosomal aberrations induced by caffeine in endoreduplicated chinese hamster cells. 20164822595
Displaying items 24901 - 25000 of 26566