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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
allosteric modulators of human a2b adenosine receptor.among adenosine receptors (ars) the a2b subtype exhibits low affinity for the endogenous agonist compared with the a1, a2a, and a3 subtypes and is therefore activated when concentrations of adenosine increase to a large extent following tissue damages (e.g. ischemia, inflammation). for this reason, a2b ar represents an important pharmacological target.201424361612
dose-rate effect was observed in t98g glioma cells following bnct.it is generally said that low let radiation produce high dose-rate effect, on the other hand, no significant dose rate effect is observed in high let radiation. although high let radiations are produced in bnct, little is known about dose-rate effect of bnct.201424360864
gain-of-function kcnh2 mutations in patients with brugada syndrome.brugada syndrome (brs) is an inherited disease characterized by right precordial st segment elevation on electrocardiograms (ecgs) that predisposes patients to sudden cardiac death as a result of polymorphic ventricular tachyarrhythmia or ventricular fibrillation (vf). in brs patients, except for scn5a, mutations in other responsible genes are poorly elucidated.201424400717
scaffold/matrix attachment regions from cho cell chromosome enhanced the stable transfection efficiency and the expression of transgene in cho cells.two scaffold/matrix attachment regions (s/mars) from chinese hamster ovary (cho) cell chromosomes were identified computationally and cloned as the domain boundaries regions. as partition chromosomes in independently regulated units to increase the exogenous gene expression, these two s/mars (mar1 and mar6) flanked expression vectors were respectively transfected into cho cells. the results showed that mar1 and mar6 were much more potent to increase the stable transfection efficiency than the ch ...201424397363
genetic variation in the two-pore domain potassium channel, task-1, may contribute to an atrial substrate for arrhythmogenesis.the two-pore domain potassium channel, k2p3.1 (task-1) modulates background conductance in isolated human atrial cardiomyocytes and has been proposed as a potential drug target for atrial fibrillation (af). task-1 knockout mice have a predominantly ventricular phenotype however, and effects of task-1 inactivation on atrial structure and function have yet to be demonstrated in vivo. the extent to which genetic variation in kcnk3, that encodes task-1, might be a determinant of susceptibility to af ...201424374141
inhibition of cardiac two-pore-domain k+ (k2p) channels by the antiarrhythmic drug vernakalant--comparison with flecainide.the mixed ion channel blocker, vernakalant (rsd1235), is effective in rapid conversion of atrial fibrillation (af) to sinus rhythm (sr). suppression of cardiac two-pore-domain potassium (k2p) channels causes action potential prolongation and has recently been proposed as a novel antiarrhythmic strategy. the objective of this study was to investigate acute effects of vernakalant on human k2p2.1 (trek-1) and k2p3.1 (task-1) channels to provide a more complete picture of its antiarrhythmic mechanis ...201424374008
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
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
xrcc2 rs3218536 polymorphism decreases the sensitivity of colorectal cancer cells to poly(adp-ribose) polymerase 1 inhibitor.single nucleotide polymorphisms (snps) are associated with the development of certain types of cancer. the present study aimed to investigate the association between x-ray repair complementing defective repair in chinese hamster cells 2 (xrcc2) snps and colorectal cancer (crc) cell sensitivity to the poly(adp-ribose) polymerase (parp) 1 inhibitor olaparib (azd2281). snapshot(®) analysis of xrcc2 snps was performed in five crc cell lines. the azd2281-sensitivities of the crc cells were also analy ...201425120693
semisynthetic model calibration for monitoring glucose in mammalian cell culture with in situ near infrared spectroscopy.near infrared (nir) spectroscopy has the capability of providing real-time, multi-analyte monitoring of the complex reaction mixture associated with cell culture processes. however, the development of robust models to predict the concentration of key analytes has proven difficult. in this study, a modeling methodology using semisynthetic process samples was used to predict glucose concentrations in chinese hamster ovary (cho) cell culture processes. partial least squares (pls) regression models ...201424284833
intracellular boron accumulation in cho-k1 cells using amino acid transport control.bpa used in bnct has a similar structure to some essential amino acids and is transported into tumor cells by amino acid transport systems. previous study groups have tried various techniques of loading bpa to increase intracellular boron concentration. cho-k1 cells demonstrate system l (lat1) activity and are suitable for specifying the transport system of a neutral amino acid. in this study, we examined the intracellular accumulation of boron in cho-k1 cells by amino acid transport control, wh ...201424388319
addition of valproic acid to cho cell fed-batch cultures improves monoclonal antibody titers.improving the productivity of a biopharmaceutical chinese hamster ovary (cho) fed-batch cell culture can enable cost savings and more efficient manufacturing capacity utilization. one method for increasing cho cell productivity is the addition of histone deacetylase (hdac) inhibitors to the cell culture process. in this study, we examined the effect of valproic acid (vpa, 2-propylpentanoic acid), a branched-chain carboxylic acid hdac inhibitor, on the productivity of three of our cho cell lines ...201424381145
mouse fyn induces pseudopodium formation in chinese hamster ovary cells.molecular mechanisms underlying the effects of fyn on cell morphology, pseudopodium movement, and cell migration were investigated. the fyn gene was subcloned into pegfp-n1 to produce pegfp-n1-fyn. chinese hamster ovary (cho) cells were transfected with pegfp-n1-fyn. the expression of fyn mrna and proteins was monitored by reverse transcription-pcr and western blotting. additionally, transfected cells were stained with 4',6-diamidino-2-phenylindole and a series of time-lapse images was taken. se ...201424378585
high-expression β(1) adrenergic receptor/cell membrane chromatography method based on a target receptor to screen active ingredients from traditional chinese medicines.β-adrenergic receptors are important targets for drug discovery. we have developed a new β1 -adrenergic receptor cell membrane chromatography (β1 ar-cmc) with offline ultra-performance lc (uplc) and ms method for screening active ingredients from traditional chinese medicines. in this study, chinese hamster ovary-s cells with high β1 ar expression levels were established and used to prepare a cell membrane stationary phase in a β1 ar-cmc model. the retention fractions were separated and identifi ...201424272943
sex hormone binding globulin and corticosteroid binding globulin as major effectors of steroid action.contrary to the long-held postulate of steroid-hormone binding globulin action, these protein carriers of steroids are major players in steroid actions in the body. this manuscript will focus on our work with sex hormone binding globulin (shbg) and corticosteroid binding globulin (cbg) and demonstrate how they are actively involved in the uptake, intracellular transport, and possibly release of steroids from cells. this manuscript will also discuss our own findings that the steroid estradiol is ...201424269740
insulin-like growth factor ii peptide fusion enables uptake and lysosomal delivery of α-n-acetylglucosaminidase to mucopolysaccharidosis type iiib fibroblasts.enzyme replacement therapy for mps iiib (mucopolysaccharidosis type iiib; also known as sanfilippo b syndrome) has been hindered by inadequate mannose 6 phosphorylation and cellular uptake of rhnaglu (recombinant human α-n-acetylglucosaminidase). we expressed and characterized a modified rhnaglu fused to the receptor-binding motif of igf-ii (insulin-like growth factor 2) (rhnaglu-igf-ii) to enhance its ability to enter cells using the cation-independent mannose 6-phosphate receptor, which is als ...201424266751
nauclea latifolia smith (rubiaceae) exerts antinociceptive effects in neuropathic pain induced by chronic constriction injury of the sciatic nerve.the roots of nauclea latifolia smith (rubiaceae) popularly known as "koumkouma" is used in traditional cameroonian medicine as neuropathic pain remedy and for the treatment of headache, inflammatory pain and convulsion. this study was conducted to evaluate the antinociceptive effects of the alkaloid fraction isolated from nauclea latifolia in neuropathic pain induced by chronic constriction injury (cci) of the sciatic nerve in rat.201424263011
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
in silico and in vitro genotoxicity evaluation of descarboxyl levofloxacin, an impurity in levofloxacin.it is important to establish the safety of impurities in drug substances or drug products. the assessment of genotoxicity of impurities and the determination of acceptable limits for genotoxic impurities was addressed in some recent guidances as a difficult issue. descarboxyl levofloxacin is an impurity isolated from levofloxacin, which may impose a risk without associated benefit. however, there is insufficient toxic information about descarboxyl levofloxacin. this study investigated the genoto ...201424224725
single-cell-based breeding: rational strategy for the establishment of cell lines from a single cell with the most favorable properties.for efficient biomolecule production (e.g., antibodies, recombinant proteins), mammalian cells with high expression rates should be selected from cell libraries, propagated while maintaining a homogenous expression rate, and subsequently stabilized at their high expression rate. clusters of isogenic cells (i.e., colonies) have been used for these processes. however, cellular heterogeneity makes it difficult to obtain cell lines with the highest expression rates by using single-colony-based breed ...201424216459
benzothiazole head group based cationic lipids: synthesis and application for gene delivery.a series of benzothiazole based lipids (1-10) containing different derivatives of benzothiazole in the head group region were synthesized to determine the structure-activity relationship for gene delivery. the liposomes formulated were mixed with plasmid dna encoding green fluorescent protein (α5gfp) or β-galactosidase (pcmv-sport-β-gal) and transfected into b16f10 (human melanoma cancer cells), cho (chinese hamster ovary), a-549 (human lung carcinoma cells) and mcf-7 (human breast carcinoma cel ...201424262373
effects of sterilization methods on the physical, chemical, and biological properties of silk fibroin membranes.silk fibroin has been widely explored for many biomedical applications, due to its biocompatibility and biodegradability. sterilization is a fundamental step in biomaterials processing and it must not jeopardize the functionality of medical devices. the aim of this study was to analyze the influence of different sterilization methods in the physical, chemical, and biological characteristics of dense and porous silk fibroin membranes. silk fibroin membranes were treated by several procedures: imm ...201424259492
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
cell line profiling to improve monoclonal antibody production.mammalian cell culture performance is influenced by both intrinsic (genetic) and extrinsic (media and process) factors. in this study, intrinsic capacity of various monoclonal antibody-producing chinese hamster ovary (cho) cell lines was compared by exposing them to the same culture condition. microarray-based transcriptomics and lc-ms/ms shotgun proteomics technologies were utilized to obtain expression landscape of different cell lines. specific transcripts and proteins correlating with produc ...201424249214
a novel approach to evaluate the pharmacokinetic biocomparability of a monoclonal antibody derived from two different cell lines using simultaneous crossover design.a parallel study design with a large number of subjects has been a typical path for pharmacokinetic (pk) biocomparability assessment of biotherapeutics with long half-lives and immunogenic propensity, for example, monoclonal antibodies (mab). a recently published innovative bioanalytical method that can quantify mab produced from two different cell lines in the same sample opened an avenue to exploring a simultaneous crossover study design for pk biocomparability assessment of biotherapeutics. s ...201424281691
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
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
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
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
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
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
in vivo and in vitro genotoxicity assessment of 2-methylisoborneol, causal agent of earthy-musty taste and odor in water.the water eutrophication process by phosphorus and nitrogen allows cyanobacteria blooms which promote, among other effects, the generation and release of the metabolite 2-methylisoborneol (2-mib) in the environment. this substance has been shown to be recalcitrant to conventional water treatment, degrading water quality. considering the limited number of studies on the biological effects of 2-mib in eukaryotic organisms, the present study assessed the genotoxicity of 2-mib using the in vitro com ...201424238741
successful construction and stable expression of an anti-cd45ra scfv-egfp fusion protein in chinese hamster ovary cells.cd45ra has been found highly expressed on leukemia cells and may be a potential target of the disease. in this study, an anti-cd45ra single-chain antibody fragment (scfv3a4) was genetically linked to the n terminus of the enhanced green fluorescent protein (egfp) to generate a scfv3a4-egfp fusion protein. the scfv3a4-egfp with a molecular weight of 57kda was stably expressed and secreted from the transfected cho cells through the er/golgi-dependent pathway. the fusion protein was soluble in the ...201424211776
γ375w fibrinogen-synthesizing cho cells indicate the accumulation of variant fibrinogen within endoplasmic reticulum.hepatic endoplasmic reticulum (er) storage disease (hersd) associated with hypofibrinogenemia has been reported in patients with four types of heterozygous γ-chain variant fibrinogen in the c terminal region. of interest, substitution of γr375w induced hypofibrinogenemia and hersd, whereas γr375g induced dysfibrinogenemia.201424210681
mammalian cell line developments in speed and efficiency.mammalian cell expression systems are the dominant tool today for producing complex biotherapeutic proteins. in this chapter, we discuss the basis for this dominance, and further explore why the chinese hamster ovary (cho) cell line has become the prevalent choice of hosts to produce most recombinant biologics. furthermore, we explore some of the innovations that are currently in development to improve the cho cell platform, from cell line specific technologies to overarching technologies that a ...201424196317
cellular responses to individual amino-acid depletion in antibody-expressing and parental cho cell lines.depletion of two nonessential amino acids, asparagine (asn) and glutamine (gln), occurred during a fed-batch production process with a cho cell line expressing a recombinant antibody. this depletion coincided with growth suppression and the onset of the stationary phase. experimental withdrawal of asn led to cell cycle arrest of cell line a in g0/g1 phase. on a mechanistic level, withdrawal of either asn or gln stimulated the amino-acid response (aar) pathway, indicating that depletion of noness ...201424254056
transgene expression from cpg-reduced lentiviral gene delivery vectors in vitro.current viral gene delivery vectors for gene therapy are inefficient due to short-lived transgene expression attributed to the cytosine-phosphate-guanine (cpg) motifs in the transgene. here we assessed the effects of cpg motif reduction in lentiviral (lv) gene delivery context on the level and duration of reporter gene expression in chinese hamster ovary (cho) cells, human immortalized myelogenous leukemia (k562) cells and hematopoietic stem cells (hscs). the cells were transduced with lv carryi ...201424120896
characterization of a pro-angiogenic, novel peptide from russell's viper (daboia russelii russelii) venom.present report shows for the first time on the induction of in vitro angiogenesis by a 3.9 kda novel peptide (rvvap) purified from russell's viper venom. secondary structure of rvvap is made up of 36.8% α-helix, 33.3% β pleated sheets and 29.9% turns. optimum angiogenesis and significant elevation in endothelial migration were observed at 50 ng/ml of rvvap treatment; above this concentration, progressive decrease in wound healing was noted. rvvap (1.0 μg/ml) was non-cytotoxic to u87-mg, hela and ...201424184153
microarray platform affords improved product analysis in mammalian cell growth studies.high throughput (ht) platforms serve as a cost-efficient and rapid screening method for evaluating the effect of cell-culture conditions and screening of chemicals. we report the development of a ht cell-based microarray platform to assess the effect of culture conditions on chinese hamster ovary (cho) cells. specifically, growth, transgene expression and metabolism of a gs/methionine sulphoximine (msx) cho cell line, which produces a therapeutic monoclonal antibody, was examined using a microar ...201424227746
arsenic trioxide preferentially induces nonapoptotic cell deaths as well as actin cytoskeleton rearrangement in the cho aa8 cell line.the therapeutic effect of arsenic trioxide (ato, as2o3) has been investigated for many years. however, the precise molecular mechanisms underlying the antitumor activity of ato are still not fully understood, but seem to depend on cell types, dosage, and duration of exposure. the purpose of this study was to assess the actin cytoskeleton rearrangement during the cell death process induced by arsenic trioxide in the cho aa8 cells. a better understanding the mechanisms of ato-action is likely to l ...201425531713
inhibition of insulin-regulated aminopeptidase (irap) by arylsulfonamides.the inhibition of insulin-regulated aminopeptidase (irap, ec 3.4.11.3) by angiotenesin iv is known to improve memory and learning in rats. screening 10 500 low-molecular-weight compounds in an enzyme inhibition assay with irap from chinese hamster ovary (cho) cells provided an arylsulfonamide (n-(3-(1h-tetrazol-5-yl)phenyl)-4-bromo-5-chlorothiophene-2-sulfonamide), comprising a tetrazole in the meta position of the aromatic ring, as a hit. analogues of this hit were synthesized, and their inhibi ...201425558444
classification of 2-pore domain potassium channels based on rectification under quasi-physiological ionic conditions.it is generally expected that 2-pore domain k(+) (k2p) channels are open or outward rectifiers in asymmetric physiological k(+) gradients, following the goldman-hodgkin-katz (ghk) current equation. although cloned k2p channels have been extensively studied, their current-voltage (i-v) relationships are not precisely characterized and previous definitions are contradictory. here we study all the functional channels from 6 mammalian k2p subfamilies in transfected chinese hamster ovary cells with p ...201425616686
[long-term cultivation of chinese hamster fibroblasts of line v-79 rjk under elevated temperature leads to karyotype structure destabilization].in this article we show that long-term cultivation of chinese hamster fibroblasts of line v-79 rjk at elevated temperature resulted in the selection of variants with genetic changes at the level of karyotype. from the first steps of resistance selection to elevated temperature we identified population of cells with changes in karyotype (polyploidy cells, deletions, inversions, translocations of chromosomes, and some cells with dm-chromosomes). further cultivation was accompanied with selection o ...201425707211
peptides derived from central turn motifs within integrin αiib and αv cytoplasmic tails inhibit integrin activation.we previously found that peptides derived from the full length of integrin αiib and αv cytoplasmic tails inhibited their parent integrin activation, respectively. here we showed that the cell-permeable peptides corresponding to the conserved central turn motif within αiib and αv cytoplasmic tails, myr-krnrppleed (αiib peptide) and myr-krvrppqeeq (αv peptide), similarly inhibited both αiib and αv integrin activation. pre-treatment with αiib or αv peptides inhibited mn(2+)-activated αiibβ3 binding ...201425290158
enhanced cellular uptake of antisecretory peptide af-16 through proteoglycan binding.peptide af-16, which includes the active site of antisecretory factor protein, has antisecretory and anti-inflammatory properties, making it a potent drug candidate for treatment of secretory and inflammatory diseases such as diarrhea, inflammatory bowel diseases, and intracranial hypertension. despite remarkable physiological effects and great pharmaceutical need for drug discovery, very little is yet understood about af-16 mechanism of action. in order to address interaction mechanisms, we inv ...201425289567
interaction of β-cyclodextrin with nile red in a single live cho cell: an initiative towards developing a prospective strategy for the excretion of adsorbed drugs from the cell membrane.a successful endeavour has been made to develop a prospective strategy to oust the drug molecules adsorbed on the cell membranes by simply using the non-toxic β-cyclodextrin (β-cd). for this purpose, fluorescent probes of different geometries and charge characteristics, namely phenosafranin (psf) and nile red (nr), were exploited for the in vitro studies using different lipids as model membranes. considering the success of the in vitro study, the present work is extended to a live chinese hamste ...201425274028
a robust transfection reagent for the transfection of cho and hek293 cells and production of recombinant proteins and lentiviral particles - ptg1.bioproduction of recombinant proteins (r-proteins) and recombinant lentiviral particles (r-lentiviral particles) requires robust transfections consisting of efficient protocols that are easy to implement, with good reproducibility for a maximum production of proteins and lentiviral particles in a short time with low cytotoxicity. this study evaluates the capacity of histidinylated polyethyleneimine i (ptg1) to facilitate robust dna transfection, with low cytotoxicity, of chinese hamster ovary (c ...201425215936
pvp-coated, negatively charged silver nanoparticles: a multi-center study of their physicochemical characteristics, cell culture and in vivo experiments.pvp-capped silver nanoparticles with a diameter of the metallic core of 70 nm, a hydrodynamic diameter of 120 nm and a zeta potential of -20 mv were prepared and investigated with regard to their biological activity. this review summarizes the physicochemical properties (dissolution, protein adsorption, dispersability) of these nanoparticles and the cellular consequences of the exposure of a broad range of biological test systems to this defined type of silver nanoparticles. silver nanoparticles ...201425383306
differential complement activation pathways promote c3b deposition on native and acetylated ldl thereby inducing lipoprotein binding to the complement receptor 1.lipoproteins can induce complement activation resulting in opsonization and binding of these complexes to complement receptors. we investigated the binding of opsonized native ldl and acetylated ldl (acldl) to the complement receptor 1 (cr1). binding of complement factors c3b, igm, c1q, mannose-binding lectin (mbl), and properdin to ldl and acldl were investigated by elisa. subsequent binding of opsonized ldl and acldl to cr1 on cr1-transfected chinese hamster ovarian cells (cho-cr1) was tested ...201425349208
lqt1 mutations in kcnq1 c-terminus assembly domain suppress iks using different mechanisms.long qt syndrome 1 (lqt1) mutations in kcnq1 that decrease cardiac iks (slowly activating delayed rectifier k(+) current) underlie ventricular arrhythmias and sudden death. lqt1 mutations may suppress iks by preventing kcnq1 assembly, disrupting surface trafficking, or inhibiting gating. we investigated mechanisms underlying how three lqt1 mutations in kcnq1 c-terminus assembly domain (r555h/g589d/l619m) decrease iks in heterologous cells and cardiomyocytes.201425344363
[expression, purification and functional identification of human psmp recombinant protein in chinese hamster ovary cells].to construct a new human chemotactic cytokine psmp eukaryotic expression vector to express psmp in chinese hamster ovary (cho) cells and to obtain the purified recombinant psmp protein for its functional mechanism study.201425331384
comparison of the biological effects of (18)f at different intracellular levels.we herein examined the biological effects of cells treated with (18)f labeled drugs for positron emission tomography (pet). the relationship between the intracellular distribution of (18)f and levels of damaged dna has yet to be clarified in detail. we used culture cells (chinese hamster ovary cells) treated with two types of (18)f labeled drugs, fluorodeoxyglucose (fdg) and fluorine ion (hf). fdg efficiently accumulated in cells, whereas hf did not. to examine the induction of dna double strand ...201425301551
peptone supplementation of culture medium has variable effects on the productivity of cho cells.the optimization of cell culture conditions for growth and productivity of recombinant chinese hamster ovary (cho) cells is a critical step in biopharmaceutical manufacturing. in the present study, the effects of the timing and amount of peptone feeding of a recombinant cho cell line grown in a basal medium in serum-free suspension culture were determined for eight peptones of different origin (plant and casein). the amino acid content and the average molecular weight of the peptones chosen were ...201425317401
inhibition of opioid receptor mediated g-protein activity after chronic administration of kynurenic acid and its derivative without direct binding to opioid receptors.there is an increasing number of evidence showing analgesic properties of the kynurenic acid (kyna), and also some studies demonstrate that kynurenine might interact with the opioid system. therefore in this study, for the first time we investigated the direct binding affinity of kyna and its structural analog kyna-1 towards mu, kappa and delta opioid receptor in competition binding experiments applying opioid receptor specific radioligands. the binding affinity measurements were performed in ch ...201425478797
direct analysis of mab aggregates in mammalian cell culture supernatant.protein aggregation during monoclonal antibody (mab) production can occur in upstream and downstream processing (dsp). current methods to determine aggregate formation during cell culture include size exclusion chromatography (sec) with a previous affinity chromatography step in order to remove disturbing cell culture components. the pre-purification step itself can already influence protein aggregation and therefore does not necessarily reflect the real aggregate content present in cell culture ...201425431119
triphlorethol-a from ecklonia cava up-regulates the oxidant sensitive 8-oxoguanine dna glycosylase 1.this study investigated the protective mechanisms of triphlorethol-a, isolated from ecklonia cava, against oxidative stress-induced dna base damage, especially 8-oxoguanine (8-oxog), in chinese hamster lung fibroblast v79-4 cells. 8-oxoguanine dna glycosylase-1 (ogg1) plays an important role in the removal of 8-oxog during the cellular response to dna base damage. triphlorethol-a significantly decreased the levels of 8-oxog induced by h2o2, and this correlated with increases in ogg1 mrna and ogg ...201425353254
protective effects of kojic acid on the periphery blood and survival of beagle dogs after exposure to a lethal dose of gamma radiation.in previous studies, it has been shown that pretreatment with kojic acid (ka) not only increased the 30 day survival rate of mice after exposed to a lethal dose of gamma radiation but also had significant radioprotective effects on the hematopoietic system, the immune system and dna of mice exposed to a 4 gy sublethal dose of radiation. furthermore, pretreatment with ka has also been shown to protect chinese hamster ovary (cho) cells against ionizing radiation-induced damage. in this investigati ...201425409121
vessel ultrasound sonographic assessment of soluble receptor for advanced glycation end products efficacy in a rat balloon injury model.we aimed to assess the therapeutic efficacy of differentially modified soluble receptor for advanced glycation end products (srage) in vivo using vessel ultrasound sonography and to compare the sonography data with those from postmortem histomorphologic analyses to have a practical reference for future clinical applications.201425408789
aristolochic acid i induces ovarian toxicity by inhibition of akt phosphorylation.aristolochic acids are natural products found in chinese herbs of the aristolochiaceae family. aristolochic acid i (aai) is a potent carcinogen and was found to be toxic in animal and clinical studies. apoptosis is a rapid, selective process of physiological cell deletion that regulates the balance between cell proliferation and cell death and is induced by various kinds of damage. however, the toxicity of aai during ovarian maturation in the mouse is unclear and is the subject of the present in ...201425406029
dual life of tppp/p25 evolved in physiological and pathological conditions.neomorphic moonlighting proteins perform distinct functions under physiological and pathological conditions without alterations at the gene level. the disordered tubulin-polymerization-promoting protein (tppp/p25), a prototype of neomorphic moonlighting proteins, modulates the dynamics and stability of the microtubule system via its bundling and tubulin acetylation-promoting activities. these physiological functions are mediated by its direct associations with tubulin/microtubules as well as tub ...201425399603
moonlighting proteins in sperm-egg interactions.sperm-egg interaction is a highly species-specific step during the fertilization process. the first steps consist of recognition between proteins on the sperm head and zona pellucida (zp) glycoproteins, the acellular coat that protects the oocyte. we aimed to determine which sperm head proteins interact with zp2, zp3 and zp4 in humans. two approaches were combined to identify these proteins: immunoblotting human spermatozoa targeted by antisperm antibodies (asas) from infertile men and far-weste ...201425399599
a high affinity recombinant antibody to the human epha3 receptor with enhanced adcc activity.epha3 is expressed in solid tumors and leukemias and is an attractive target for the therapy. we have generated a panel of humaneered® antibodies to the ligand-binding domain using a fab epitope-focused library that has the same specificity as monoclonal antibody miiia4. a high-affinity antibody was selected that competes with the miiia4 antibody for binding to epha3 and has an improved affinity of ∼1 nm. in order to generate an antibody with potent cell-killing activity the variable regions wer ...201425413948
dimer-dependent intrinsic/basal activity of the class b g protein-coupled receptor pac1 promotes cellular anti-apoptotic activity through wnt/β-catenin pathways that are associated with dimer endocytosis.the high expression of pacap (pituitary adenylate cyclase-activating polypeptide)-preferring receptor pac1 is associated with nerve injury and tumors. our previous report (yu r, et al. plos one 2012; 7: e51811) confirmed the dimerization of pac1 and found that the m-pac1 mutation in the n-terminal first cys/ala lost the ability to form dimers. in this study, chinese hamster ovary (cho-k1) cells overexpressing wild-type pac1 (pac1-cho) had significantly higher anti-apoptotic activities against se ...201425426938
monitoring the dielectric response of single cells following mitochondrial adenosine triphosphate synthase inhibition by oligomycin using a dielectrophoretic cytometer.one of the main uses of adenosine triphosphate (atp) within mammalian cells is powering the na(+)/k(+) atpase pumps used to maintain ion concentrations within the cell. since ion concentrations determine the cytoplasm conductivity, atp concentration is expected to play a key role in controlling the cytoplasm conductivity. the two major atp production pathways within cells are via glycolysis within the cytoplasm and via the electron transport chain within the mitochondria. in this work, a differe ...201425553191
[establishment and application of human cho/ntr1 system].neurotensin receptor-1 (ntr1), which can stimulate the intracellular cascade signal pathway, belongs to the large superfamily of g-protein coupled receptors. ntr1 is related to the occurrence and development of several kinds of diseases. in order to screen the inhibitors for the cancers associated with ntr1 protein, we established a cho (chinese hamster ovary) cell line in which human neurotensin receptor-1 was highly expressed. the method is to construct the recombinant plasmid which was lysed ...201425518326
antiarrhythmic efficacy of cpuy102122, a multiple ion channel blocker, on rabbits with ischemia/reperfusion injury.the antiarrhythmic potential of a novel multichannel blocker cpuy102122 (cy22) was investigated in the present study.201425443730
cyto-genotoxicity assessment of potential radioprotector, 3,3'-diselenodipropionic acid (dsepa) in chinese hamster ovary (cho) cells and human peripheral blood lymphocytes.our previous study showed that 3,3'-diselenodipropionic acid (dsepa), a simple, stable, and water-soluble organoselenium exhibiting glutathione peroxidase (gpx)-like activity offered good radioprotection under in vitro and in vivo conditions. herein, we investigated the anti-genotoxic effect of dsepa in model cellular systems such as chinese hamster ovary (cho) cell line and human peripheral lymphocytes after exposure to γ-radiation. the measurements on the induction of γ-h2ax foci and micronucl ...201425440905
monoclonal antibodies biosimilarity assessment using transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry.out of all categories, monoclonal antibody (mab) therapeutics attract the most interest due to their strong therapeutic potency and specificity. six of the 10 top-selling drugs are antibody-based therapeutics that will lose patent protection soon. the european medicines agency has pioneered the regulatory framework for approval of biosimilar products and approved the first biosimilar antibodies by the end of 2013. as highly complex glycoproteins with a wide range of micro-variants, mabs require ...201425484058
rapid identification of an antibody dna construct rearrangement sequence variant by mass spectrometry.during cell line development for an igg1 antibody candidate (mab1), a c-terminal extension was identified in 2 product candidate clones expressed in cho-k1 cell line. the extension was initially observed as the presence of anomalous new peaks in these clones after analysis by cation exchange chromatography (cex-hplc) and reduced capillary electrophoresis (rce-sds). reduced mass analysis of these cho-k1 clones revealed that a larger than expected mass was present on a sub-population of the heavy ...201425484040
a pan-cancer analysis of alternative splicing events reveals novel tumor-associated splice variants of matriptase.high-throughput transcriptome sequencing allows identification of cancer-related changes that occur at the stages of transcription, pre-messenger rna (mrna), and splicing. in the current study, we devised a pipeline to predict novel alternative splicing (as) variants from high-throughput transcriptome sequencing data and applied it to large sets of tumor transcriptomes from the cancer genome atlas (tcga). we identified two novel tumor-associated splice variants of matriptase, a known cancer-asso ...201425506199
xrcc2 promotes colorectal cancer cell growth, regulates cell cycle progression, and apoptosis.x-ray repair complementing defective repair in chinese hamster cells 2 (xrcc2) and poly(adp-ribose) polymerase 1 (parp1) both play important roles in homologous recombination dna repair. according to the theory of synthetic lethality, xrcc2-deficient cells are more sensitive to parp1 inhibitors compared to xrcc2-expressing cells. we investigated xrcc2 expression and function in colorectal cancer (crc), and the characteristics of sensitivity to parp1 inhibitor in crc cells with different xrcc2 le ...201425526472
enhanced susceptibility of ovaries from obese mice to 7,12-dimethylbenz[a]anthracene-induced dna damage.7,12-dimethylbenz[a]anthracene (dmba) depletes ovarian follicles and induces dna damage in extra-ovarian tissues, thus, we investigated ovarian dmba-induced dna damage. additionally, since obesity is associated with increased offspring birth defect incidence, we hypothesized that a dmba-induced dna damage response (ddr) is compromised in ovaries from obese females. wild type (lean) non agouti (a/a) and kk.cg-ay/j heterozygote (obese) mice were dosed with sesame oil or dmba (1mg/kg; intraperitone ...201425448685
human cyp2e1-dependent and human sulfotransferase 1a1-modulated induction of micronuclei by benzene and its hydroxylated metabolites in chinese hamster v79-derived cells.benzene is a ubiquitous environmental pollutant and a confirmed human carcinogen, which requires metabolic activation, primarily by cyp2e1, for most of its biological actions. chromosome damages in benzene-exposed workers and rodents have been observed, and in their urine sulfo- and glucuronide-conjugates of phenol and hydroquinone were present. yet, direct evidence for human cyp2e1-activated mutagenicity of benzene and the exact significance of phase ii metabolism for inactivating benzene metab ...201425771868
effects of nutrient levels and average culture ph on the glycosylation pattern of camelid-humanized monoclonal antibody.the impact of operating conditions on the glycosylation pattern of humanized camelid monoclonal antibody, eg2-hfc produced by chinese hamster ovary (cho) cells has been evaluated by a combination of experiments and modeling. cells were cultivated under different levels of glucose and glutamine concentrations with the goal of investigating the effect of nutrient depletion levels and ammonia build up on the cell growth and the glycoprofiles of the monoclonal antibody (mab). the effect of average p ...201425014402
cloning and expression of the functional human anti-vascular endothelial growth factor (vegf) using the pcdna3.1 vector and the human chronic myelogenous leukemia cell line k562.in this study, the light chain (κ) and heavy chain (γ) sequences of the monoclonal antibody against vascular endothelial growth factor (vegf) were sub-cloned into the eukaryotic pcdna3.1 (+) (hygro) and the pcdna3.1 (+) (neo) expression vectors using the traditional and homologous recombination methods. to express the antibody, the recombinant plasmids were transfected into the chinese hamster ovary (cho) and the k562 cell lines. the recombinant antibody was then purified using the protein a aff ...201424399318
mitochondria represent another locale for the divalent metal transporter 1 (dmt1).the divalent metal transporter (dmt1) is well known for its roles in duodenal iron absorption across the apical enterocyte membrane, in iron efflux from the endosome during transferrin-dependent cellular iron acquisition, as well as in uptake of non-transferrin bound iron in many cells. recently, using multiple approaches, we have obtained evidence that the mitochondrial outer membrane is another subcellular locale of dmt1 expression. while iron is of vital importance for mitochondrial energy me ...201425483589
serum-free medium optimization based on trial design and support vector regression.the plackett-burman design and support vector machine (svm) were reported to be used on many fields such as some feature selections, protein structure prediction, or forecasting of other situations. here, with suspension adapted chinese hamster ovary (cho) cells as the object of study, a serum-free medium for the culture of cho cells in suspension was optimized by this method. support vector machine based on genetic algorithm was used to predict the growth rate of cho and prove the results from ...201425379507
characterization of full-length recombinant human proteoglycan 4 as an ocular surface boundary lubricant.proteoglycan 4 (prg4, or lubricin) is a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role. recent technological advances have enabled abundant expression of full-length recombinant human prg4 (rhprg4). the objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhprg4, and 2) assess the ocular surface boundary lubricating abili ...201424997456
rakkyo fructan as a cryoprotectant for serum-free cryopreservation of mammalian cells.cryopreservation refers to the long-term storage of mammalian cells. mammalian serum is generally used as a cryoprotectant, but is associated with problems including the risk of contamination by pathogens and quality control issues. therefore, a serum-free cryopreservation method needs to be established. in this study, we focused on rakkyo fructan, a fructose polymer, derived from the japanese shallot as an alternative factor to serum. fructan contributes to tolerance to frost and dehydration in ...201424485744
comparison of in vitro toxicity of mainstream cigarette smoke particulate matter from nano- to micro-size.in order to investigate the interrelationship between particulate matter (pm) size and in vitro toxicological effects of mainstream cigarette smoke, pm sized between 10 nm and 10 μm in mainstream cigarette smoke was sampled and divided into six stages. the in vitro cytotoxicity, genotoxicity and cell inhibition effects of pm were assessed by the neutral red cytotoxicity assay, salmonella mutagenicity assay, micronucleus test and flow cytometry analysis, respectively. the results showed that all ...201424333866
the role of the 3' region of mammalian gonadotropin β subunit gene in the luteinizing hormone to chorionic gonadotropin evolution.cgβ subunits comprise a unique carboxyl-terminal peptide (ctp) that has multiple o-linked glycans and extends serum half-life of the protein. it has evolved by incorporating a previously untranslated region of the lhβ gene into the reading frame. although ctp-like sequences are encrypted in the lhβ genes of several mammals, the cgβ subunit developed only in primates and equids. to study this restriction in evolution, we examined whether the cryptic ctp decoded from the bovine lhβ gene (boctp) po ...201424239648
absence of micronucleus formation in cho-k1 cells cultivated in platelet lysate enriched medium.human platelet lysate (pl) represents an effective substitute of fetal bovine serum (fbs) for mesenchymal stromal cell (msc) cultivation. compared to fbs, pl favors msc proliferation significantly shortening the population doubling time and avoiding the risks related to the use of animal derivatives. growth factors contained in the platelets are released upon platelet disruption following freezing/thawing cycles or as we have recently described by using ultrasound. we have investigated whether t ...201424290702
conventional and whitening toothpastes: cytotoxicity, genotoxicity and effect on the enamel surface.to evaluate the cytotoxicity and genotoxicity of whitening and common toothpastes, and the surface roughness of tooth enamel submitted to brushing with both toothpastes.201425707084
human non-pigmented ciliary epithelium bradykinin b2-receptors: receptor localization, pharmacological characterization of intracellular ca(2+) mobilization, and prostaglandin secretion.to characterize the bradykinin (bk) receptor system in human non-pigmented ciliary epithelium (npce) using immunohistochemistry and functional cell-based techniques.201424624903
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