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tubb1 mutation disrupting microtubule assembly impairs proplatelet formation and results in congenital macrothrombocytopenia.this report describes a family with tubb1-associated macrothrombocytopenia diagnosed based on abnormal platelet β1-tubulin distribution. a circumferential marginal microtubule band was undetectable, whereas microtubules were frayed and disorganized in every platelet from the affected individuals. patients were heterozygous for novel tubb1 p.f260s that locates at the α- and β-tubulin intradimer interface. mutant β1-tubulin was not incorporated into microtubules with endogenous α-tubulin, and α-tu ...201424344610
calcium electroporation in three cell lines: a comparison of bleomycin and calcium, calcium compounds, and pulsing conditions.electroporation with calcium (calcium electroporation) can induce atp depletion-associated cellular death. in the clinical setting, the cytotoxic drug bleomycin is currently used with electroporation (electrochemotherapy) for palliative treatment of tumors. calcium electroporation offers several advantages over standard treatment options: calcium is inexpensive and may readily be applied without special precautions, as is the case with cytostatic drugs. therefore, details on the use of calcium e ...201424342489
low ph inactivation for xenotropic gamma retrovirus in recombinant human tnf-α receptor immunoglobulin g and mechanism of inactivation.cho-derived recombinant proteins for human therapeutic are used commonly. there are noninfectious endogenous retroviruses in cho cells. validation study for inactivation process is required. murine xenotropic gamma retrovirus (x-mulv) is a model virus in validation study. in our previous study, optimum conditions for x-mulv inactivation were sifted. in this study, we performed a further research on low ph inactivation for evaluation of x-mulv clearance in manufacturing of recombinant human tnf-α ...201424341971
asparagine-linked glycosylation of bone morphogenetic protein-2 is required for secretion and osteoblast differentiation.bone morphogenetic protein-2 (bmp-2), a glycosylated protein, has been demonstrated to play a key role in osteoblast differentiation. however, the function of its glycosylation is incompletely understood. in this study, we examined the role that n-linked glycans (nlg) play in the secretion of bmp-2. blocking the addition of nlgs or inhibiting initial glycan processing prevented the secretion of bmp-2. to identify the specific glycosylation sites, we abolished potential sites of n-linked glycosyl ...201424336949
digital mrna profiling of n-glycosylation gene expression in recombinant chinese hamster ovary cells treated with sodium butyrate.to understand the effects of sodium butyrate (nabu) on protein glycosylation, recombinant chinese hamster ovary (rcho) cells producing fc-fusion glycoprotein were subjected to 3mm nabu. the addition of nabu to the cultures reduced the relative proportion of acidic isoforms and sialic acid content of the glycoprotein. fifty-two n-glycosylation-related gene expressions were also assessed by the nanostring ncounter system, which can provide a direct digital readout using custom-designed color-coded ...201424333461
a novel lysophosphatidic acid acyltransferase enzyme (lpaat4) with a possible role for incorporating docosahexaenoic acid into brain glycerophospholipids.glycerophospholipids are important components of cellular membranes, required for constructing structural barriers, and for providing precursors of bioactive lipid mediators. lysophosphatidic acid acyltransferases (lpaats) are enzymes known to function in the de novo glycerophospholipid biosynthetic pathway (kennedy pathway), using lysophosphatidic acid (lpa) and acyl-coa to form phosphatidic acid (pa). until now, three lpaats (lpaat1, 2, and 3) have been reported from the 1-acyl-glycerol-3-phos ...201424333445
low concentration of exogenous carbon monoxide protects mammalian cells against proliferation induced by radiation-induced bystander effect.radiation-induced bystander effect (ribe) has been proposed to have tight relationship with the irradiation-caused secondary cancers beyond the irradiation-treated area after radiotherapy. our previous studies demonstrated a protective effect of low concentration carbon monoxide (co) on the genotoxicity of ribe after α-particle irradiation. in the present work, a significant inhibitory effect of low-dose exogenous co, generated by tricarbonyldichlororuthenium (ii) dimer [co-releasing molecule (c ...201424333162
virus-free transient protein production in sf9 cells.a method for virus-free transient gene expression from suspension-adapted sf9 insect cells was developed with the gene of interest being expressed from a plasmid carrying the homologous region 5 enhancer (hr5) and the immediate early 1 (ie1) promoter from autographa californica multicapsid nucleopolyhedrovirus (acmnpv). under the optimal conditions described in the study, cells were transfected at a density of 30×10⁶ cells/ml with 0.9 μg dna and 1.35 μg of linear 25 kd polyethylenimine (pei) per ...201424333123
bipolar nanosecond electric pulses are less efficient at electropermeabilization and killing cells than monopolar pulses.multiple studies have shown that bipolar (bp) electric pulses in the microsecond range are more effective at permeabilizing cells while maintaining similar cell survival rates as compared to monopolar (mp) pulse equivalents. in this paper, we investigated whether the same advantage existed for bp nanosecond-pulsed electric fields (nspef) as compared to mp nspef. to study permeabilization effectiveness, mp or bp pulses were delivered to single chinese hamster ovary (cho) cells and the response of ...201424332942
synthesis and biological evaluation of novel 2-amino-3-aroyl-4-neopentyl-5-substituted thiophene derivatives as allosteric enhancers of the a₁ adenosine receptor.2-amino-3-benzoyl thiophenes have been widely reported to act as allosteric enhancers at the a1 adenosine receptor. their activity can be increased considerably by appropriate substitutions at the 4- and 5-positions of the thiophene ring. substituent size at the thiophene c-4 position seemed to be a factor closely related to activity, with the 4-neopentyl (2,2-dimethylpropyl) substitution showing the greatest enhanced activity. a wide series of 2-amino-3-aroyl-4-neopentylthiophene derivatives wi ...201424332652
glycosylation analysis of an aggregated antibody produced by chinese hamster ovary cells in bioreactor culture.n-glycosylation of therapeutic antibodies contributes not only to their biological function, but also to their stability and tendency to aggregate. here, we investigated the impact of the glycosylation status of an aggregated antibody that accumulated during the bioreactor culture of chinese hamster ovary cells. high-performance liquid chromatography analysis showed that there was no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibo ...201424326352
analysis of dna repair gene polymorphisms in glioblastoma.glioblastoma is the most common and aggressive primary brain tumor in adults. despite several factors such as ionizing radiation exposure or rare genetic syndromes have been associated with the development of glioblastoma, no underlying cause has been identified for the majority of cases. we thus aimed to investigate the role of dna repair polymorphisms in modulating glioblastoma risk.201424325908
endogenous microrna clusters outperform chimeric sequence clusters in chinese hamster ovary cells.micrornas (mirnas) are small non-coding rnas (∼22 nucleotides) which regulate gene expression by silencing mrna translation. mirnas are transcribed as long primary transcripts, which are enzymatically processed by drosha/dgcr8, in the nucleus, and by dicer in the cytoplasm, into mature mirnas. the importance of mirnas for coordinated gene expression is commonly accepted. consequentially, there is a growing interest in the application of mirnas to improve phenotypes of mammalian cell factories su ...201424323929
evaluation of a peg/hydroxypropyl starch aqueous two-phase system for the separation of monoclonal antibodies from cell culture supernatant.in this study, an aqueous two-phase system (atps) with peg and hydroxypropyl starch (hps) was used to separate monoclonal antibody (mab) from chinese hamster ovary cell culture supernatant. the phase diagram of the peg/hps atps was determined, and the effects of nacl addition were investigated. the results showed that nacl addition could lead to a shift of the binodal curve and that phase separation would occur at higher peg and hps concentrations. the effects of nacl addition, ph, and the load ...201424318171
natural and glucosyl flavonoids inhibit poly(adp-ribose) polymerase activity and induce synthetic lethality in brca mutant cells.poly(adp-ribose) polymerase (parp) inhibitors have been proven to represent superior clinical agents targeting dna repair mechanisms in cancer therapy. we investigated parp inhibitory effects of the natural and synthetic flavonoids (quercetin, rutin, monoglucosyl rutin and maltooligosyl rutin) and tested the synthetic lethality in brca2 mutated cells. in vitro elisa assay suggested that the flavonoids have inhibitory effects on parp activity, but glucosyl modifications reduced the inhibitory eff ...201424317580
trehalose suppresses antibody aggregation during the culture of chinese hamster ovary cells.the aggregation of therapeutic antibodies during the manufacturing process is problematic because of the potential risks posed by the aggregates, such as an unexpected immune response. one of the hallmark effects of trehalose, a disaccharide consisting of two alpha-glucose units, is as a chemical chaperone with anti-aggregation activity. in this study, chinese hamster ovary (cho) cell line producing a diabody-type bispecific antibody were cultured in medium containing trehalose and the aggregati ...201424315529
predicting the expression of recombinant monoclonal antibodies in chinese hamster ovary cells based on sequence features of the cdr3 domain.despite the development of high-titer bioprocesses capable of producing >10 g l(-1) of recombinant monoclonal antibody (mab), some so called "difficult-to-express" (dte) mabs only reach much lower process titers. for widely utilized "platform" processes the only discrete variable is the protein coding sequence of the recombinant product. however, there has been little systematic study to identify the sequence parameters that affect expression. this information is vital, as it would allow us to r ...201424311306
disposable bioreactors for inoculum production and protein expression.disposable bioreactors have been increasingly implemented over the past ten years. this relates to both r & d and commercial manufacture, in particular, in animal cell-based processes. among the numerous disposable bioreactors which are available today, wave-mixed bag bioreactors and stirred bioreactors are predominant. whereas wave-mixed bag bioreactors represent the system of choice for inoculum production, stirred systems are often preferred for protein expression. for this reason, the author ...201424297422
¹h-nmr protocol for exometabolome analysis of cultured mammalian cells.(1)h-nuclear magnetic resonance ((1)h-nmr) spectroscopy is a powerful technique to analyze the composition of complex mixtures based on the particular proton fingerprint of each molecule. here we describe a protocol for exometabolome analysis of mammalian cells using this technique, including sample preparation, spectra acquisition, and integration. the potential of this technique is exemplified by application to cultures of a chinese hamster ovary (cho) cell line. the average error associated t ...201424297420
screening and optimization of chemically defined media and feeds with integrated and statistical approaches.the majority of therapeutic proteins are expressed in mammalian cells, predominantly in chinese hamster ovary cells. while cell culture media and feed supplements are crucial to protein productivity, medium optimization can be labor intensive and time-consuming. in this chapter, we describe some basic concepts in medium development and introduce a rational and rapid workflow to screen and optimize media and feeds. the major goal of medium screening is to select a base formulation as the foundati ...201424297413
high-throughput synchronization of mammalian cell cultures by spiral microfluidics.the development of mammalian cell cycle synchronization techniques has greatly advanced our understanding of many cellular regulatory events and mechanisms specific to different phases of the cell cycle. in this chapter, we describe a high-throughput microfluidic-based approach for cell cycle synchronization. by exploiting the relationship between cell size and its phase in the cell cycle, large numbers of synchronized cells can be obtained by size fractionation in a spiral microfluidic channel. ...201424297405
genotoxic mixtures and dissimilar action: concepts for prediction and assessment.combinations of genotoxic agents have frequently been assessed without clear assumptions regarding their expected (additive) mixture effects, often leading to claims of synergisms that might in fact be compatible with additivity. we have shown earlier that the combined effects of chemicals, which induce micronuclei (mn) in the cytokinesis-block micronucleus assay in chinese hamster ovary-k1 cells by a similar mechanism, were additive according to the concept of concentration addition (ca). here, ...201424297155
mechanistic evaluation of the transfection barriers involved in lipid-mediated gene delivery: interplay between nanostructure and composition.here we present a quantitative mechanism-based investigation aimed at comparing the cell uptake, intracellular trafficking, endosomal escape and final fate of lipoplexes and lipid-protamine/deoxyribonucleic acid (dna) (lpd) nanoparticles (nps) in living chinese hamster ovary (cho) cells. as a model, two lipid formulations were used for comparison. the first formulation is made of the cationic lipid 1,2-dioleoyl-3-trimethylammonium-propane (dotap) and the zwitterionic lipid dioleoylphosphocholine ...201424296066
aspects of nonviral gene therapy: correlation of molecular parameters with lipoplex structure and transfection efficacy in pyridinium-based cationic lipids.this study seeks correlations between the molecular structures of cationic and neutral lipids, the lipid phase behavior of the mixed-lipid lipoplexes they form with plasmid dna, and the transfection efficacy of the lipoplexes. synthetic cationic pyridinium lipids were co-formulated (1:1) with the cationic lipid 1,2-dimyristoyl-sn-glycero-3-ethylphosphocholine (epc), and these lipids were co-formulated (3:2) with the neutral lipids 1,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine (dope) or chol ...201424296044
dynamic kisspeptin receptor trafficking modulates kisspeptin-mediated calcium signaling.kisspeptin receptor (kiss1r) signaling plays a critical role in the regulation of reproduction. we investigated the role of kisspeptin-stimulated kiss1r internalization, recycling, and degradation in the modulation of kiss1r signaling. kisspeptin stimulation of chinese hamster ovary or gt1-7 cells expressing kiss1r resulted in a biphasic increase in intracellular ca(2+) ([ca(2+)]i), with a rapid acute increase followed by a more sustained second phase. in contrast, stimulation of the trh recepto ...201424295737
analysis of chinese hamster ovary cell metabolism through a combined computational and experimental approach.optimization of cell culture processes can benefit from the systematic analysis of experimental data and their organization in mathematical models, which can be used to decipher the effect of individual process variables on multiple outputs of interest. towards this goal, a kinetic model of cytosolic glucose metabolism coupled with a population-level model of chinese hamster ovary cells was used to analyse metabolic behavior under batch and fed-batch cell culture conditions. the model was parame ...201424292563
risk assessment of polychlorinated biphenyls and heavy metals in soils of an abandoned e-waste site in china.risk assessment of abandoned e-waste recycling areas received little attention. herein, we report the concentrations of 16 pcbs and 7 heavy metals in soils near an abandoned e-waste recycling plant in taizhou, china. our data showed that levels of tri-, tetra-, penta-, hexa-pcbs were 9.01, 5.56, 12.93, 3.13 mg/kg, and pb, cd, cu were 6082.9, 42.3, 2364.2 mg/kg soil. cd was the most prevalent contaminant with nemerow index value of 44.3. contaminants have been transported from the abandoned site ...201424292442
solvation dynamics and intermittent oscillation of cell membrane: live chinese hamster ovary cell.dynamics of the exofacial thiols (i.e., cell surface thiol containing membrane proteins) of a live chinese hamster ovary (cho) cell is probed by time-resolved confocal microscopy. for this purpose, a fluorescent probe, 7-(diethylamino)-3-(4-maleimidophenyl)-4-methylcoumarin (cpm) is covalently attached to the exofacial thiols. the emission maximum of cpm bound exofacial thiols indicates a highly exposed and polar environment. using cpm, we studied solvation dynamics, for the first time, at the m ...201424571625
heat exposure enhances radiosensitivity by depressing dna-pk kinase activity during double strand break repair.from the role of double strand dna dependent protein kinase (dna-pkcs) activity of non-homologous end joining (nhej) repair for dna double strand breaks (dsbs), we aim to define possible associations between thermo-sensitisation and the enzyme activities in x-ray irradiated cells.201424571173
mir-7 inhibits colorectal cancer cell proliferation and induces apoptosis by targeting xrcc2.analysis using publicly available algorithms predicts that x-ray repair complementing defective repair in chinese hamster cells 2 (xrcc2), a key component in the homologous recombination repair pathway, is a potential target of micro-ribonucleic acid-7 (mir-7). some studies have shown that both mir-7 and xrcc2 are associated with cancer development. for this purpose, we searched for the possible relationship between mir-7 and xrcc2 in the development of colorectal cancer (crc).201424570594
apoptosis-inducing activity of hplc fraction from voacanga globosa (blanco) merr. on the human colon carcinoma cell.voacanga globosa (blanco), a plant endemic to the philippines, is traditionally used especially by indigenous people of bataan in the treatment of ulcers, wounds and tumorous growths. this study aimed to provide scientific evidence to therapeutic properties by determining cytotoxic and pro-apoptotic activity of hplc fractions from leaves on hct116 human colon carcinoma and a549 human lung carcinoma cell lines. ethanolic extraction was performed on v globosa leaves followed by hexane and ethyl ac ...201424568467
improved expression of recombinant human factor ix by co-expression of ggcx, vkor and furin.recombinant human fix concentrates (rhfix) are essential in the treatment and prevention of bleeding in the bleeding disorder haemophilia b. however, due to the complex nature of fix production yields are low which leads to high treatment costs. here we report the production of rhfix with substantially higher yield by co-expressing human fix with ggcx (γ-glutamyl carboxylase), vkor (vitamin k epoxide reductase) and furin (paired basic amino acid cleaving enzyme) in chinese hamster ovary (cho) ce ...201424567122
[study on the effect of deficient ercc2/xpd gene on the repair of dna damage induced by uvc in cho cell line].to explore the function of ercc2/xpd in the repair of dna damage induced by uvc.201424564106
a phase i/ii, multiple-dose, dose-escalation study of siltuximab, an anti-interleukin-6 monoclonal antibody, in patients with advanced solid tumors.this phase i/ii study evaluated safety, efficacy, and pharmacokinetics of escalating, multiple doses of siltuximab, a chimeric anti-interleukin (il)-6 monoclonal antibody derived from a new chinese hamster ovary (cho) cell line in patients with advanced/refractory solid tumors.201424563479
neonatal genome-wide methylation patterns in relation to birth weight in the norwegian mother and child cohort.although epigenetic regulation plays a critical role in embryonic development, few studies have examined the relationship of epigenome-wide methylation with fetal growth. using the infinium humanmethylation450 beadchip (illumina, inc., san diego, california) in a substudy of 1,046 infants from the norwegian mother and child cohort study (moba) enrolled between 1999 and 2008, we examined epigenome-wide cord blood dna methylation in relation to birth weight. in multivariable-adjusted robust linear ...201424561991
the impact of homologous recombination repair deficiency on depleted uranium clastogenicity in chinese hamster ovary cells: xrcc3 protects cells from chromosome aberrations, but increases chromosome fragmentation.depleted uranium (du) is extensively used in both industry and military applications. the potential for civilian and military personnel exposure to du is rising, but there are limited data on the potential health hazards of du exposure. previous laboratory research indicates du is a potential carcinogen, but epidemiological studies remain inconclusive. du is genotoxic, inducing dna double strand breaks, chromosome damage and mutations, but the mechanisms of genotoxicity or repair pathways involv ...201424561002
long-term activation upon brief exposure to xanomleline is unique to m1 and m4 subtypes of muscarinic acetylcholine receptors.xanomeline is an agonist endowed with functional preference for m1/m4 muscarinic acetylcholine receptors. it also exhibits both reversible and wash-resistant binding to and activation of these receptors. so far the mechanisms of xanomeline selectivity remain unknown. to address this question we employed microfluorometric measurements of intracellular calcium levels and radioligand binding to investigate differences in the short- and long-term effects of xanomeline among muscarinic receptors expr ...201424558448
effect of sodium butyrate on the assembly, charge variants, and galactosylation of antibody produced in recombinant chinese hamster ovary cells.sodium butyrate (nabu) is known to increase the specific productivity of recombinant chinese hamster ovary (rcho) cells. to understand the effects of nabu on the product quality, rcho cells producing monoclonal antibody (mab) were cultivated at various concentrations of nabu (0 to 4 mm). nabu increased correctly assembled mab. in the absence of nabu, the proportions of intact mab (2h2l) and heavy chain dimer (2h) were 81 and 15 %. at 1 mm nabu, the proportion of 2h2l increased to 93 %, whereas t ...201424557571
sodium butyrate enhances the acidic isoform content of recombinant human erythropoietin produced by chinese hamster ovary cells.sodium butyrate is commonly used in mammalian cell cultures to increase the productivity of recombinant proteins. a chinese hamster ovary (cho) cell line producing recombinant human erythropoietin (rhepo) was cultured in commercial medium. addition of 0.5 mm butyrate inhibited the over-growth of the cells after the medium was changed from serum-added medium to serum-free medium. at the 6th day, the addition of butyrate lowered the transcriptional level of sialidases i, ii, and iii compared to th ...201424557070
expression of base excision repair key factors and mir17 in familial and sporadic breast cancer.understanding of brca1/2 interaction with the base excision repair (ber) pathway could improve therapy based on 'synthetic lethality', whose effectiveness is based on homologous recombination deficiency in cells lacking functional brca genes. however, poly (adp-ribose) polymerase (parp) inhibitors failed in some patients and for this reason we explored ber key enzyme expression. in this study, the expression of ber enzymes (redox factor 1/apurinic-apyrimidinic endonuclease 1 (ref1/apex1), nth en ...201424556691
profiling convoluted single-dimension proton nmr spectra: a plackett-burman approach for assessing quantification error of metabolites in complex mixtures with application to cell culture.single-dimension hydrogen, or proton, nuclear magnetic resonance spectroscopy (1d-(1)h nmr) has become an attractive option for characterizing the full range of components in complex mixtures of small molecular weight compounds due to its relative simplicity, speed, spectral reproducibility, and noninvasive sample preparation protocols compared to alternative methods. one challenge associated with this method is the overlap of nmr resonances leading to "convoluted" spectra. while this can be mit ...201424555717
a strategy for fusion expression and preparation of functional glucagon-like peptide-1 (glp-1) analogue by introducing an enterokinase cleavage site.kglp-1, a 31-amino acid glucagon-like peptide-1 (glp-1) analogue, has a great therapeutic potential for anti-diabetes. in this work, a strategy for expression and purification of functional kglp-1 peptide has been established. kglp-1 cdna was fused with glutathione s-transferase (gst), with an enterokinase cleavage site in the fusion junction. the recombinant fusion protein gst-kglp-1 was affinity purified via the gst-tag, and then digested with enterokinase. the resulting gst part as well as th ...201424737080
efficient production and purification of extracellular domain of human fgfr-fc fusion proteins from chinese hamster ovary cells.the family of fibroblast growth factor receptors (fgfrs) plays an important role in cell growth, survival, differentiation and angiogenesis. the three immunoglobulin-like extracellular domains of fgfr (d1, d2, and d3) are critical for ligand binding and specificity towards fibroblast growth factor and heparan sulfate. fibroblast growth factor receptors are overexpressed in a wide variety of tumors, such as breast, bladder, and prostate cancer, and therefore they are attractive targets for differ ...201424727156
optimization of heterologous protein production in chinese hamster ovary cells under overexpression of spliced form of human x-box binding protein.the optimization of protein production is a complex and challenging problem in biotechnology. different techniques for transcription, translation engineering and the optimization of cell culture conditions have been used to improve protein secretion, but there remain many open problems involving post-translational modifications of the secreted protein and cell line stability.201424725707
effect of glutamine substitution by tca cycle intermediates on the production and sialylation of fc-fusion protein in chinese hamster ovary cell culture.in an effort to reduce the accumulation of ammonia in culture medium, three different tca cycle intermediates, (alpha-ketoglutarate (α-kg), citric acid and succinic acid) along with glutamic acid for a comparison, were examined as a substitute for glutamine with rcho cell line producing a fc-fusion glycoprotein. among them, α-kg showed the best production performance. when cells were cultivated with 4 mm α-kg, the final ammonia concentration did not exceed 3 mm, which is less than one fourth of ...201424721212
ultrasound driven assembly of lignin into microcapsules for storage and delivery of hydrophobic molecules.oil-filled microcapsules of kraft lignin were synthesized by first creating an oil in water emulsion followed by a high-intensity, ultrasound-assisted cross-linking of lignin at the water/oil interface. the rationale behind our approach is based on promoting documented lignin hydrophobic interactions within the oil phase, followed by locking the resulting spherical microsystems by covalent cross-linking using a high intensity ultrasound treatment. as further evidence in support of our rationale, ...201424720505
synthesis, spectral characterization, and antiproliferative studies of mixed ligand titanium complexes of adamantylamine.titanium complexes have been synthesized by the reaction between titanium tetrachloride (ticl4), respective bidentate ligand [4,4' -dimethoxy-2,2' -bipyridine (bpome), 6,6'-dimethyl-2,2'-bipyridine (dpme), 1,2-diaminocyclohexane (dach), 1,10-phenanthroline (phen), and benzoylacetone (bzac)], and adamantylamine (ada) in 1 : 2 : 2 molar ratios, respectively. the structure of synthesized complexes was confirmed using elemental analysis, ftir, uv-visible, (1)h nmr, and mass spectrometry techniques. ...201424715822
surfactant uptake dynamics in mammalian cells elucidated with quantitative coherent anti-stokes raman scattering microspectroscopy.the mechanism of surfactant-induced cell lysis has been studied with quantitative coherent anti-stokes raman scattering (cars) microspectroscopy. the dynamics of surfactant molecules as well as intracellular biomolecules in living chinese hamster lung (chl) cells has been examined for a low surfactant concentration (0.01 w%). by using an isotope labeled surfactant having cd bonds, surfactant uptake dynamics in living cells has been traced in detail. the simultaneous cars imaging of the cell itse ...201424710120
modulation of amyloid precursor protein expression reduces β-amyloid deposition in a mouse model.proteolytic cleavage of the amyloid precursor protein (app) generates β-amyloid (aβ) peptides. prolonged accumulation of aβ in the brain underlies the pathogenesis of alzheimer disease (ad) and is regarded as a principal target for development of disease-modifying therapeutics.201424687915
evaluation of anti-inflammatory and mechanism of action of extract of macrosiphonia longiflora (desf.) müll. arg.macrosiphonia longiflora (desf.) müll. arg (apocynaceae), popularly known as 'velame' and 'velame branco', is a native subshrub that grows in the brazilian cerrado. this plant is widely used in traditional medicine in the form of decoction and infusion, particularly as anti-inflammatory, depurative, anti-rheumatic, antisyphilitic and antiulcer remedy. there is no available information in the literature that has addressed its pharmacological activity and phytochemical analysis.201424681039
microrna-9 and microrna-326 regulate human dopamine d2 receptor expression, and the microrna-mediated expression regulation is altered by a genetic variant.the human dopamine receptor d2 (drd2) has been implicated in the pathophysiology of schizophrenia and other neuropsychiatric disorders. most antipsychotic drugs influence dopaminergic transmission through blocking dopamine receptors, primarily drd2. we report here the post-transcriptional regulation of drd2 expression by two brain-expressed micrornas (mirs), mir-326 and mir-9, in an ex vivo mode, and show the relevance of mir-mediated drd2 expression regulation in human dopaminergic neurons and ...201424675081
rorα and ror γ are expressed in human skin and serve as receptors for endogenously produced noncalcemic 20-hydroxy- and 20,23-dihydroxyvitamin d.rorα and rorγ are expressed in human skin cells that produce the noncalcemic 20-hydroxyvitamin d3 [20(oh)d3] and 20,23-dihydroxyvitamin d3 [20,23(oh)2d3]. chinese hamster ovary (cho) cells stably expressing a tet-on rorα or rorγ expression vector and a ror-responsive element (rore)-luc reporter, and a mammalian 2-hybrid model examining the interaction between the ligand binding domain (lbd) of rorα or rorγ with an lbd-interacting lxxll-peptide, were used to study ror-antagonist activities. these ...201424668754
a high throughput screening assay system for the identification of small molecule inhibitors of gsp.mis-sense mutations in the α-subunit of the g-protein, gsα, cause fibrous dysplasia of bone/mccune-albright syndrome. the biochemical outcome of these mutations is constitutively active gsα and increased levels of camp. the aim of this study was to develop an assay system that would allow the identification of small molecule inhibitors specific for the mutant gsα protein, the so-called gsp oncogene. commercially available chinese hamster ovary cells were stably transfected with either wild-type ...201424667240
setting time affects in vitro biological properties of root canal sealers.biocompatibility of root canal sealers is important because of the long-term contact of their eluates and/or degradation products with periapical tissues. the literature still lacks studies about the genotoxic effects of these materials and the influence of setting time on biological properties. the cytotoxicity and genotoxicity of an epoxy resin-based sealer (ah plus), a single methacrylate-based sealer (endorez), and a silicone-based sealer (roekoseal) were assessed.201424666905
a real-time method for measuring camp production modulated by gαi/o-coupled metabotropic glutamate receptors.group ii and group iii metabotropic glutamate (mglu) receptors are g protein-coupled receptors (gpcrs) that inhibit adenylyl cyclase via activation of gαi/o. the purpose of this study was to design a universal method that overcomes previous challenges in consistently measuring group ii and group iii mglu-receptor (mglur) activation in stably transfected systems. in chinese hamster ovary (cho) cells stably transfected with the glosensor camp biosensor, we optimized conditions for simple and highl ...201424659805
confocal imaging with a fluorescent bile acid analogue closely mimicking hepatic taurocholate disposition.this study aimed to characterize the in vitro hepatic transport mechanisms in primary rat and human hepatocytes of the fluorescent bile acid derivative n-(24-[7-(4-n,n-dimethylaminosulfonyl-2,1,3-benzoxadiazole)]amino-3α,7α,12α-trihydroxy-27-nor-5β-cholestan-26-oyl)-2'-aminoethanesulfonate (tauro-nor-thca-24-dbd), previously synthesized to study the activity of the bile salt export pump (bsep). the fluorescent bile acid derivative exhibited saturable uptake kinetics in suspended rat hepatocytes. ...201424652646
role of oatp-1b1 and/or oatp-1b3 in hepatic disposition of tyrosine kinase inhibitors.the metabolism of tyrosine kinase inhibitors (tkis) is mainly mediated via hepatic route, but the mechanism responsible for their hepatocellular accumulation is still unknown. this study was designed to understand the contribution of organic anion transporting polypeptides (oatps) in the hepatic uptake of selected tkis - pazopanib, canertinib, erlotinib, vandetanib and nilotinib.201424643910
efficient folding/assembly in chinese hamster ovary cells is critical for high quality (low aggregate content) of secreted trastuzumab as well as for high production: stepwise multivariate regression analyses.when developing cell culture processes for therapeutic antibodies, the low content of aggregated proteins is the most critical because administering aggregated antibody molecules might result in adverse effects such as immunogenicity. to characterize cells with high productivity and quality, we determined factors that are closely related to antibody titer, which is a productivity indicator, and the area percentage of high molecular weight species in cultivated media, which is equivalent to aggre ...201424635945
association study of c.910a>g and c.1686c>g polymorphisms in xrcc1 gene with risk of hepatocellular carcinoma in the chinese population.xrcc1 (human x-ray repair complementing defective repair in chinese hamster cell 1) gene is considered a potentially important gene influencing the risk of hepatocellular carcinoma (hcc). our analyses detected two allelic variants of xrcc1, c.910a>g and c.1686c>g. we aimed to investigate whether these polymorphisms influence the risk of hcc. the association between the xrcc1 polymorphisms and the risk of hcc was analyzed in 719 patients and 662 controls by polymerase chain reaction-restriction f ...201424634229
pathogenic strains of acanthamoeba are recognized by tlr4 and initiated inflammatory responses in the cornea.free-living amoebae of the acanthamoeba species are the causative agent of acanthamoeba keratitis (ak), a sight-threatening corneal infection that causes severe pain and a characteristic ring-shaped corneal infiltrate. innate immune responses play an important role in resistance against ak. the aim of this study is to determine if toll-like receptors (tlrs) on corneal epithelial cells are activated by acanthamoeba, leading to initiation of inflammatory responses in the cornea. human corneal epit ...201424633052
analyzing clonal variation of monoclonal antibody-producing cho cell lines using an in silico metabolomic platform.monoclonal antibody producing chinese hamster ovary (cho) cells have been shown to undergo metabolic changes when engineered to produce high titers of recombinant proteins. in this work, we have studied the distinct metabolism of cho cell clones harboring an efficient inducible expression system, based on the cumate gene switch, and displaying different expression levels, high and low productivities, compared to that of the parental cells from which they were derived. a kinetic model for cho cel ...201424632968
pharmacological characterization of endomorphin-2-based cyclic pentapeptides with methylated phenylalanine residues.as part of our continuing studies on the structure-activity relationships of cyclic pentapeptides based on the structure of endomorphin-2, we report here the synthesis and biological activities of a new series of analogs incorporating 2', 3' or 4'-methylphenylalanine (mephe) residues into positions 3 or 4 of the parent cyclopeptide, dmt-c[d-lys-phe-phe-asp]nh2 (dmt=2',6'-dimethyltyrosine). analogs with mephe in position 4 showed a row of magnitude increased μ-opioid receptor (mop receptor) affin ...201424632335
a novel luminescence-based method for the detection of functionally active antibodies to muscarinic acetylcholine receptors of the m3 type (machr3) in patients' sera.in different bioassays, functional antibodies reacting with the human muscarinic acetylcholine receptor m3(machr3) have been detected in sera from patients with sjögren's syndrome (ss), and there is strong evidence that those antibodies may have pathogenetic relevance. however, depending on the method of detection, their prevalence varied. furthermore, those bioassays are difficult to standardize. we report on the development and optimization of a novel test system based on a luminometric method ...201424628494
the brca2 gene is a potential molecular target during 5-fluorouracil therapy in human oral cancer cells.5-fluorouracil (5-fu) is widely used in clinical cancer therapy. it is commonly used either alone or in combination with other drugs and/or radiation for head and neck, and other types of cancers. 5-fu induces dna double-strand breaks (dsbs). inhibition of the repair of 5-fu-induced dsbs may improve the therapeutic response in many tumors to this anticancer agent. the aim of the present study was to further our understanding of the pathways which are involved in the repair of 5-fu-induced dsbs. ...201424627042
disruption of the actin cortex contributes to susceptibility of mammalian cells to nanosecond pulsed electric fields.nanosecond pulsed electric fields (nspefs) perturb membranes of cultured mammalian cells in a dose-dependent manner with different types of cells exhibiting characteristic survivability. adherent cells appear more robust than non-adherent cells during whole-cell exposure. we hypothesize that cellular elasticity based upon the actin cytoskeleton is a contributing parameter, and the alteration of a cell's actin cortex will significantly affect viability upon nspef exposure. chinese hamster ovary ( ...201424619788
cell compatible arginine containing cationic polymer: one-pot synthesis and preliminary biological assessment.synthetic cationic polymers are of interest as both nonviral vectors for intracellular gene delivery and antimicrobial agents. for both applications synthetic polymers containing guanidine groups are of special interest since such kind of organic compounds/polymers show a high transfection potential along with antibacterial activity. it is important that the delocalization of the positive charge of the cationic group in guanidine significantly decreases the toxicity compared to the ammonium func ...201424619618
turoctocog alfa for the treatment of hemophilia a.patients with hemophilia a lack functional coagulation factor viii (fviii), causing excessive spontaneous bleeding episodes and bleeding during trauma or surgery. plasma-derived or synthetic recombinant fviii is used to control bleeding, but currently available treatments are limited because of patient development of fviii inhibitors and cross-contamination of human and animal-derived infectious agents. turoctocog alfa is a recombinant fviii produced in chinese hamster ovary cells that has demon ...201424619589
high-throughput screening platform for engineered nanoparticle-mediated genotoxicity using cometchip technology.the likelihood of intentional and unintentional engineered nanoparticle (enp) exposure has dramatically increased due to the use of nanoenabled products. indeed, enps have been incorporated in many useful products and have enhanced our way of life. however, there are many unanswered questions about the consequences of nanoparticle exposures, in particular, with regard to their potential to damage the genome and thus potentially promote cancer. in this study, we present a high-throughput screenin ...201424617523
transcriptome analyses of cho cells with the next-generation microarray cho41k: development and validation by analysing the influence of the growth stimulating substance igf-1 substitute longr(3.).the increasing importance of chinese hamster ovary (cho) cells for the production of pharmaceutical proteins has awakened the demand to understand the cellular metabolism of these cells. however, satisfactory gene expression studies have yet been impractical due to insufficient coverage of sequences. in this work, previously determined sequence information of cho cells and newly derived data from 454 and illumina sequencing was used to establish the cho41k microarray which contains 41,304 probes ...201424613301
presenilin-1 influences processing of the acetylcholinesterase membrane anchor prima.presenilin-1 (ps1) is the catalytic component of the γ-secretase complex. in this study, we explore if ps1 participates in the processing of the cholinergic acetylcholinesterase (ache). the major ache variant expressed in the brain is a tetramer (g(4)) bound to a proline-rich membrane anchor (prima). overexpression of the transmembrane prima protein in chinese hamster ovary cells expressing ache and treated with the γ-secretase inhibitor n-[n-(3,5-difluorophenacetyl)-l-alanyl]-s-phenylglycine t- ...201424612677
occurrence and subcellular distribution of the nadphx repair system in mammals.hydration of nad(p)h to nad(p)hx, which inhibits several dehydrogenases, is corrected by an atp-dependent dehydratase and an epimerase recently identified as the products of the vertebrate carkd (carbohydrate kinase domain) and aibp (apolipoprotein ai-binding protein) genes respectively. the purpose of the present study was to assess the presence of these enzymes in mammalian tissues and determine their subcellular localization. the carkd gene encodes proteins with a predicted mitochondrial prop ...201424611804
influence of the phase effect on gradient-based and statistics-based focus measures in bright field microscopy.autofocusing is essential to high throughput microscopy and live cell imaging and requires reliable focus measures. phase objects such as separated single chinese hamster ovary cells are almost invisible at the optical focus position in bright field microscopy images. because of the phase effect, defocused images of phase objects have more contrast. in this paper, we show that widely used focus measures exhibit an untypical behaviour for such images. in the case of homogeneous cells, that is, wh ...201424611652
heat-transfer resistance measurement method (htm)-based cell detection at trace levels using a progressive enrichment approach with highly selective cell-binding surface imprints.surface-imprinted polymers allow for specific cell detection based on simultaneous recognition of the cell shape, cell size, and cell membrane functionalities by macromolecular cell imprints. in this study, the specificity of detection and the detection sensitivity for target cells within a pool of non-target cells were analyzed for a cell-specific surface-imprinted polymer combined with a heat-transfer-based read-out technique (htm). a modified chinese hamster ovarian cell line (cho-ldld) was u ...201424606112
promiscuous gating modifiers target the voltage sensor of k(v)7.2, trpv1, and h(v)1 cation channels.some of the fascinating features of voltage-sensing domains (vsds) in voltage-gated cation channels (vgccs) are their modular nature and adaptability. here we examined the vsd sensitivity of different vgccs to 2 structurally related nontoxin gating modifiers, nh17 and nh29, which stabilize k(v)7.2 potassium channels in the closed and open states, respectively. the effects of nh17 and nh29 were examined in chinese hamster ovary cells transfected with transient receptor potential vanilloid 1 (trpv ...201424599966
genotoxic evaluation of titanium dioxide nanoparticles in vivo and in vitro.with the extensive application of titanium dioxide (tio2) nanoparticles (nps) in food industry, there is a rising debate concerning the possible risk associated with exposure to tio2 nps. the purpose of this study is to evaluate the genotoxicity of tio2 nps using in vivo and in vitro test systems. in vivo study, the adult male sprague-dawley rats were exposed to anatase tio2 nps (75 ± 15 nm) through intragastric administration at 0, 10, 50 and 200mg/kg body weight every day for 30 days. the γ-h2 ...201424594277
safety and mutagenicity evaluation of red mold dioscorea fermented from monascus purpureus ntu 568.monascus-fermented products, including red mold rice and red mold dioscorea, have been developed as functional foods with many health benefits. we performed safety and mutagenic evaluations on red mold dioscorea powder (rmdp) fermented from monascus purpureus ntu 568. the results of ames test using salmonella typhimurium strains ta97a, ta98, ta100, ta102, and ta1535 showed that rmdp (⩽5 mg/plate) was not mutagenic. the mammalian chromosomal aberration test showed that the number of chinese hamst ...201424582683
poly-γ-glutamic acid enhances the quality of recombant erythropoietin produced by cho cells.the effect of poly-γ-glutamic acid (γpga), which is produced by bacillus sp., on the production of recombinant erythropoietin (repo) by chinese hamster ovary (cho) cells in suspension culture was evaluated. the growth, viability, and productivity of recombinant cho cells were investigated in a chemically defined medium with 50 and 500 kd γpgas at 0.075% or with pluronic f68. cell growth with the two γpgas was lower than that with pluronic f68 but significantly higher than that without any additi ...201426019520
cellular and genomic toxicity produced by uv light in chinese hamster ovary cells.uvb and uvc toxicity was detected in chinese hamster ovary (cho) cell lines aa8, uv5 and xem2 (a v79-derived cell line expressing rat p450 1a1). unlike ficz-hplc assay that showed induction of cyp1a1 enzyme activity after 20 minutes and 2 hour uvc exposure, the erod assay showed no difference in cytochrome p450 1a1 (cyp1a1) activity after exposure to different doses of uvb and uvc light. different cytotoxic and mutagenic effect of photo lesions induced by uvc and uvb light was investigated with ...201424577918
efficiency improvement of an antibody production process by increasing the inoculum density.increasing economic pressure is the main driving force to enhance the efficiency of existing processes. we developed a perfusion strategy for a seed train reactor to generate a higher inoculum density for a subsequent fed batch production culture. a higher inoculum density can reduce culture duration without compromising product titers. hence, a better capacity utilization can be achieved. the perfusion strategy was planned to be implemented in an existing large scale antibody production process ...201424574274
antibodies that detect o-linked β-d-n-acetylglucosamine on the extracellular domain of cell surface glycoproteins.the transfer of n-acetylglucosamine (glcnac) to ser or thr in cytoplasmic and nuclear proteins is a well known post-translational modification that is catalyzed by the o-glcnac transferase ogt. a more recently identified o-glcnac transferase, eogt, functions in the secretory pathway and transfers o-glcnac to proteins with epidermal growth factor-like (egf) repeats. a number of antibodies that detect o-glcnac in cytosolic and nuclear extracts have been described previously. here we compare seven ...201424573683
arp2, a novel pro-apoptotic protein expressed in epithelial prostate cancer lncap cells and epithelial ovary cho transformed cells.neoplastic epithelial cells generate the most aggressive types of cancers such as those located in the lung, breast, colon, prostate and ovary. during advanced stages of prostate cancer, epithelial cells are associated to the appearance of androgen-independent tumors, an apoptotic-resistant phenotype that ultimately overgrows and promotes metastatic events. we have previously identified and electrophysiologically characterized a novel ca(2+)-permeable channel activated during apoptosis in the an ...201424465888
high-throughput miniaturized bioreactors for cell culture process development: reproducibility, scalability, and control.decreasing the timeframe for cell culture process development has been a key goal toward accelerating biopharmaceutical development. advanced microscale bioreactors (ambr™) is an automated micro-bioreactor system with miniature single-use bioreactors with a 10-15 ml working volume controlled by an automated workstation. this system was compared to conventional bioreactor systems in terms of its performance for the production of a monoclonal antibody in a recombinant chinese hamster ovary cell li ...201424449637
mutations in pgap3 impair gpi-anchor maturation, causing a subtype of hyperphosphatasia with mental retardation.glycosylphophatidylinositol (gpi)-anchored proteins play important roles in many biological processes, and mutations affecting proteins involved in the synthesis of the gpi anchor are reported to cause a wide spectrum of intellectual disabilities (ids) with characteristic additional phenotypic features. here, we describe a total of five individuals (from three unrelated families) in whom we identified mutations in pgap3, encoding a protein that is involved in gpi-anchor maturation. three sibling ...201424439110
construction of a public cho cell line transcript database using versatile bioinformatics analysis pipelines.chinese hamster ovary (cho) cell lines represent the most commonly used mammalian expression system for the production of therapeutic proteins. in this context, detailed knowledge of the cho cell transcriptome might help to improve biotechnological processes conducted by specific cell lines. nevertheless, very few assembled cdna sequences of cho cells were publicly released until recently, which puts a severe limitation on biotechnological research. two extended annotation systems and web-based ...201424427317
very-long-chain polyunsaturated fatty acids accumulate in phosphatidylcholine of fibroblasts from patients with zellweger syndrome and acyl-coa oxidase1 deficiency.peroxisomes are subcellular organelles that function in multiple anabolic and catabolic processes, including β-oxidation of very-long-chain fatty acids (vlcfa) and biosynthesis of ether phospholipids. peroxisomal disorders caused by defects in peroxisome biogenesis or peroxisomal β-oxidation manifest as severe neural disorders of the central nervous system. abnormal peroxisomal metabolism is thought to be responsible for the clinical symptoms of these diseases, but their molecular pathogenesis r ...201424418004
new melatonin (mt1/mt2) ligands: design and synthesis of (8,9-dihydro-7h-furo[3,2-f]chromen-1-yl) derivatives.herein we describe the synthesis of novel tricyclic analogues issued from the rigidification of the methoxy group of the benzofuranic analogue of melatonin as mt1 and mt2 ligands. most of the synthesized compounds displayed high binding affinities at mt1 and mt2 receptors subtypes. compound 6b (mt1, ki=0.07nm; mt2, ki=0.08nm) exhibited with the vinyl 6c and allyl 6d the most interesting derivatives of this series. functional activity of these compounds showed full agonist activity with ec50 in t ...201424417958
soluble receptor for advanced glycation end products (srage) attenuates haemodynamic changes to chronic hypoxia in the mouse.the calgranulin-like protein mts1/s100a4 and the receptor for advanced glycation end-products (rage) have recently been implicated in mediating pulmonary arterial smooth muscle cell proliferation and vascular remodelling in experimental pulmonary arterial hypertension (ph). here, the effects of rage antagonism upon 2 weeks of hypobaric hypoxia (10% o2)-induced ph in mice were assessed. treatment with srage was protective against hypobaric hypoxia-induced increases in right ventricular pressure b ...201424417910
cytoskeleton modification and cholesterol depletion affect membrane properties and caveolae positioning of cho cells.the formation of protrusions is necessary for numerous biological processes. it involves extension of the plasma membrane, and the force needed for this is provided by the actin cytoskeleton. tether pulling with optical tweezers can mimic the formation of a protrusion, so we used this method to investigate the effects of modifying not only actin (with latrunculin a) but also microtubules (with nocodazole) and the plasma membrane itself (with methyl-β-cyclodextrin) on the chinese hamster ovary ce ...201424413749
analysis of inter-subunit contacts reveals the structural malleability of extracellular domains in platelet glycoprotein ib-ix complex.the glycoprotein (gp)ib-ix complex is critical to hemostasis and thrombosis. its proper assembly is closely correlated with its surface expression level and requires cooperative interactions among extracellular and transmembrane domains of ibα, ibβ and ix subunits. two interfaces have been previously identified between the extracellular domains of ibβ and ix.201424406065
[construction and expression of a eukaryotic expression vector containing human cr2-fc fusion protein].to construct a eukaryotic expression vector containing human complement receptor 2 (cr2)-fc and express the cr2-fc fusion protein in chinese hamster ovary (cho) cells.201424405588
polyamine transport inhibitors: design, synthesis, and combination therapies with difluoromethylornithine.the development of polyamine transport inhibitors (ptis), in combination with the polyamine biosynthesis inhibitor difluoromethylornithine (dfmo), provides a method to target cancers with high polyamine requirements. the dfmo+pti combination therapy results in sustained intracellular polyamine depletion and cell death. a series of substituted benzene derivatives were evaluated for their ability to inhibit the import of spermidine in dfmo-treated chinese hamster ovary (cho) and l3.6pl human pancr ...201424405276
influence of npc1 genotype on the toxicity of hydroxypropyl-β-cyclodextrin, a potentially therapeutic agent, in niemann-pick type c disease models.hydroxypropyl-β-cyclodextrin (hpbcd) is an attractive drug candidate against niemann-pick type c (npc) disease. however, the safety of hpbcd treatment for npc patients remains to be elucidated. in this study, we examined the acute toxicity of hpbcd in npc1-deficient mice. when treated with hpbcd (20,000 mg/kg, subcutaneously), over half of the wild-type (npc1(+/+)) or npc1(+/-) mice died by 72 h after the injection. in contrast, all of the npc1(-/-) mice survived. marked pathophysiological chang ...201427896072
comprehensive, quantitative bioprocess productivity monitoring using fluorescence eem spectroscopy and chemometrics.this study demonstrates the application of fluorescence excitation-emission matrix (eem) spectroscopy to the quantitative predictive analysis of recombinant glycoprotein production cultured in a chinese hamster ovary (cho) cell fed-batch process. the method relies on the fact that eem spectra of complex solutions are very sensitive to compositional change. as the cultivation progressed, changes in the emission properties of various key fluorophores (e.g., tyrosine, tryptophan, and the glycoprote ...201424504094
pecam-1-dependent heme oxygenase-1 regulation via an nrf2-mediated pathway in endothelial cells.the antioxidant enzyme heme oxygenase (ho)-1, which catalyses the first and rate-limiting step of heme degradation, has major anti-inflammatory and immunomodulatory effects via its cell-type-specific functions in the endothelium. in the current study, we investigated whether the key endothelial adhesion and signalling receptor pecam-1 (cd31) might be involved in the regulation of ho-1 gene expression in human endothelial cells (ecs). to this end pecam-1 expression was down-regulated in human umb ...201424500083
controllability analysis of protein glycosylation in cho cells.to function as intended in vivo, a majority of biopharmaceuticals require specific glycan distributions. however, achieving a precise glycan distribution during manufacturing can be challenging because glycosylation is a non-template driven cellular process, with the potential for significant uncontrolled variability in glycan distributions. as important as the glycan distribution is to the end-use performance of biopharmaceuticals, to date, no strategy exists for controlling glycosylation on-li ...201424498415
apolipoprotein e increases cell association of amyloid-β 40 through heparan sulfate and lrp1 dependent pathways.the increased risk of alzheimer's disease (ad) associated with specific apolipoprotein e (apoe) isoforms appears to relate to altered amyloid-β (aβ) homeostasis. clearance of aβ from the brain is reduced in the presence of the ad-associated apoe4 isoform, which may contribute to the accumulation of aβ deposits in the parenchyma and vasculature. the low-density lipoprotein receptor-related protein 1 (lrp1) and heparan sulfate proteoglycans (hspgs), both established apoe receptors, are involved in ...201424491019
analysis of microrna transcription and post-transcriptional processing by dicer in the context of cho cell proliferation.cho cells are the mammalian cell line of choice for recombinant production of therapeutic proteins. however, their low rate of proliferation limits obtainable space-time yields due to inefficient biomass accumulation. we set out to correlate microrna transcription to cell-specific growth-rate by microarray analysis of 5 cho suspension cell lines with low to high specific growth rates. global microrna expression analysis and pearson correlation studies showed that mature microrna transcript level ...201424486028
long-term episomal transgene expression from mitotically stable integration-deficient lentiviral vectors.nonintegrating gene delivery vectors have an improved safety profile compared with integrating vectors, but transgene retention is problematic as nonreplicating episomes are progressively and rapidly diluted out through cell division. we have developed an integration-deficient lentiviral vector (idlv) system generating mitotically stable episomes capable of long-term transgene expression. we found that a transient cell cycle arrest at the time of transduction with idlvs resulted in 13-45% of chi ...201424483952
comparison of clot lysis activity and biochemical properties of originator tenecteplase (metalyse(®)) with those of an alleged biosimilar.the bioengineered tissue plasminogen activator tenecteplase is an important treatment modality of acute myocardial infarction recommended by international guidelines. following introduction of originator tenecteplase (brand names metalyse(®) and tnkase(®)), a "biosimilar" tenecteplase became available for commercial use in india under the brand name elaxim(®) in the absence of indian biosimilar guidelines which came into force from september 15th, 2012. based on a report of biochemical and fibri ...201424550831
human orexin/hypocretin receptors form constitutive homo- and heteromeric complexes with each other and with human cb1 cannabinoid receptors.human ox1 orexin receptors have been shown to homodimerize and they have also been suggested to heterodimerize with cb1 cannabinoid receptors. the latter has been suggested to be important for orexin receptor responses and trafficking. in this study, we wanted to assess the ability of the other combinations of receptors to also form similar complexes. vectors for expression of human ox1, ox2 and cb1 receptors, c-terminally fused with either renilla luciferase or gfp(2) green fluorescent protein ...201424530395
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