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Selected AbstractsRare species loss alters ecosystem function , invasion resistanceECOLOGY LETTERS, Issue 4 2001Kelly G. Lyons The imminent decline in species diversity coupled with increasing exotic species introductions has provoked investigation into the role of resident diversity in community resistance to exotic species colonization. Here we present the results of a field study using an experimental method in which diversity was altered by removal of less abundant species and the resulting disturbance was controlled for by removal of an equivalent amount of biomass of the most common species from paired plots. Following these manipulations, the exotic grass, Lolium temulentum, was introduced. We found that exotic species establishment was higher in plots in which diversity was successfully reduced by removal treatments and was inversely related to imposed species richness. These results demonstrate that less common species can significantly influence invasion events and highlight the potential role of less common species in the maintenance of ecosystem function. [source] Synthesis, Complexation and Spectrofluorometric Studies of a New NS3 Anthracene-Containing Macrocyclic LigandEUROPEAN JOURNAL OF INORGANIC CHEMISTRY, Issue 15 2006Abel Tamayo Abstract A new fluorescent device for detecting protons and metal ions, 11-(9-anthracenylmethyl)-1,4,7-trithia-11-azacyclotetradecane (L), has been synthesised. In addition, the photophysical properties of both the free and protonated species have been examined by absorption and fluorescence titrations of dichloromethane solutions of L with methanesulfonic acid. The coordinating properties of L toward PdII, ZnII, NiII and CoII have been studied both in solution and in the solid state. Different behaviours have been observed in the absorption and fluorescence titrations of L with the above-mentioned transition-metal ions. To evaluate whether these differences were due to the existence of equilibria between protonated and complexed species, such titrations have been repeated in the presence of an equivalent amount of acid. The structure of the [Pd(L)](BF4)2 complex has been solved by X-ray crystallography. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2006) [source] A Versatile Solvent-Free "One-Pot" Route to Polymer Nanocomposites and the in situ Formation of Calcium Phosphate/Layered Silicate Hybrid NanoparticlesADVANCED FUNCTIONAL MATERIALS, Issue 11 2010Hans Weickmann Abstract Poly(methyl methacrylate) (PMMA), polystyrene (PS), and polyurethane (PU) nanocomposites containing well-dispersed calcium phosphate/layered silicate hybrid nanoparticles were prepared in a versatile solvent-free "one-pot" process without requiring separate steps, such as organophilic modification, purification, drying, dispersing, and compounding, typical for many conventional organoclay nanocomposites. In this "one-pot" process, alkyl ammonium phosphates were added as swelling agents to a suspension of calcium/layered silicate in styrene, methyl methacrylate, or polyols prior to polymerization. Alkyl ammonium phosphates were prepared in situ by reacting phosphoric acid with an equivalent amount of alkyl amines such as stearyl amine (SA) or the corresponding ester- and methacrylate-functionalized tertiary alkyl amines, obtained via Michael Addition of SA with methyl acrylate or ethylene 2-methacryloxyethyl acrylate. Upon contact with the calcium bentonite suspension, the cation exchange of Ca2+ in the silicate interlayers for alkyl ammonium cations rendered the bentonite organophilic and enabled effective swelling in the monomer accompanied by intercalation and in situ precipitation of calcium phosphates. According to energy dispersive X-ray analysis, the calcium phosphate precipitated exclusively onto the surfaces of the bentonite nanoplatelets, thus forming easy-to-disperse calcium phosphate/layered silicate hybrid nanoparticles. Incorporation of 5,15,wt% of such hybrid nanoparticles into PMMA, PS, and PU afforded improved stiffness/toughness balances of the polymer nanocomposites. Functionalized alkyl ammonium phosphate addition enabled polymer attachment to the nanoparticle surfaces. Transmission electron microscopy (TEM) analyses of PU and PU-foam nanocomposites, prepared by dispersing hybrid nanoparticles in the polyols prior to isocyanate cure, revealed the formation of fully exfoliated hybrid nanoparticles. [source] Are Selling, General, and Administrative Costs "Sticky"?JOURNAL OF ACCOUNTING RESEARCH, Issue 1 2003Mark C. Anderson A fundamental assumption in cost accounting is that the relation between costs and volume is symmetric for volume increases and decreases. In this study, we investigate whether costs are "sticky",that is, whether costs increase more when activity rises than they decrease when activity falls by an equivalent amount. We find, for 7,629 firms over 20 years, that selling, general, and administrative (SG&A) costs increase on average 0.55% per 1% increase in sales but decrease only 0.35% per 1% decrease in sales. Our analysis compares the traditional model of cost behavior in which costs move proportionately with changes in activity with an alternative model in which sticky costs occur because managers deliberately adjust the resources committed to activities. We test hypotheses about the properties of sticky costs and how the degree of stickiness of SG&A costs varies with firm circumstances. [source] Sodium-induced calcium deficiency in sugar beet during substitution of potassium by sodiumJOURNAL OF PLANT NUTRITION AND SOIL SCIENCE, Issue 2 2009Abdul Wakeel Abstract Functions of sodium (Na+) and potassium (K+) are closely associated. In some crops, Na+ is able to prevent or reduce considerably the occurrence of K+ deficiency. Sugar beet (Beta vulgaris L.) is a natrophilic crop, and positive effects of Na+ applications on yield were observed when K+ was sufficiently supplied. However, it is not known which specific function of K+ can limit the growth of sugar beet when K+ is substituted by an equivalent amount of Na+. Therefore, K+ substitution by Na+ was investigated for sugar beet in hydroponics. Surprisingly, no K+ -deficiency symptoms were observed. However, calcium (Ca2+) concentrations in the leaves were significantly decreased. Moreover, Ca2+ uptake and translocation through xylem sap were reduced in Na+ -treated plants. It is concluded that Ca2+ uptake by roots and its translocation via xylem sap primarily limit the possibility of K+ substitution by Na+ in sugar beet nutrition. [source] Effects of Light and Dark Beer on Hepatic Cytochrome P-450 Expression in Male Rats Receiving Alcoholic Beverages as Part of Total Enteral NutritionALCOHOLISM, Issue 5 2005Mats Hidestrand Background: Alcoholic beverages contain many congeners in addition to ethanol. Therefore, consumption of alcoholic beverages may have considerably different effects on expression of hepatic microsomal monooxygenases than the relatively selective induction of cytochrome P-450 (CYP) 2E1 observed after consumption of pure ethanol. Methods: In the current study, we compared the effects of two beers: lager (a light roasted beer) and stout (a dark roasted beer) with those of an equivalent amount of pure ethanol on hepatic CYP expression. Beer or pure ethanol was part of a complete total enteral nutrition diet that was infused intragastrically into male Sprague Dawley rats for 21 days. At the end of the infusion period, rats were euthanized, and liver and intestinal microsomes were prepared. Expression and activity of CYP1A1/2, CYP2B1, CYP2E1, CYP3A, and CYP4A were assessed by Western immunoblotting and by using CYP enzyme,specific substrates, respectively. Results: mRNA and protein levels of CYP4A1 were elevated only in stout-treated animals. However, lauric acid 12-hydroxylase activity (a CYP4A-specific activity) was reduced (p, 0.05) in microsomes from lager- and stout-fed rats. After oxidation with potassium ferricyanide, this activity was significantly increased in microsomes from stout-fed animals. The relative expression of CYP2E1 and CYP2B1 and the activities toward p -nitrophenol, pentoxyresorufin, or benzyloxyresorufin did not differ between beers or compared with pure ethanol or controls. However, the mean expression of CYP1A2, CYP3A, and CYP4A apoproteins was greater in liver microsomes from stout-infused rats than in those from lager-infused rats, ethanol-infused rats, and diet controls (p, 0.05). In addition, although no significant differences were observed in ethoxyresorufin O-dealkylase (EROD), methoxyresorufin O-dealkylase (MROD), midazolam, or testosterone hydroxylase activities between groups, stout-infused rats had greater hepatic microsomal erythromycin N -demethylase activity than other groups (p, 0.05). Conclusions: Stout contains components other than ethanol that interact in a complex fashion with the monooxygenase system. [source] The effect of Anethum graveolens L. (dill) on corticosteroid induced diabetes mellitus: involvement of thyroid hormonesPHYTOTHERAPY RESEARCH, Issue 12 2008Sunanda Panda Abstract An investigation was made to evaluate the role of Anethum graveolens L. (dill) leaf extract in the regulation of corticosteroid-induced type 2 diabetes mellitus in female rats. In dexamethasone-treated animals (1 mg/kg for 22 days) an increase in serum concentration of insulin and glucose and in hepatic lipid peroxidation (LPO) was observed. However, there was a decrease in serum concentration of thyroid hormones and in the endogenous antioxidant enzymes, such as superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) in liver. In animals treated with an equivalent amount of dexamethasone for a similar period (22 days) when received the leaf extract (100 mg/kg b.wt/d.) for last 15 days a decrease in the concentration of both serum glucose and insulin was observed, indicating the potential of the plant extract in the regulation of corticosteroid-induced diabetes. Dexamethasone-induced alterations in the levels of thyroid hormones as well as in hepatic LPO, SOD, CAT and GSH were also reversed by the plant extract. Copyright © 2008 John Wiley & Sons, Ltd. [source] Antioxidative effects of pumpkin seed (Cucurbita pepo) protein isolate in CCl4-Induced liver injury in low-protein fed ratsPHYTOTHERAPY RESEARCH, Issue 11 2006C. Z. Nkosi Abstract The effects of pumpkin seed (Cucurbita pepo) protein isolate on the plasma activity levels of catalase (CA), superoxide dismutase (SOD), glutathione peroxidase (GSHpx) and total antioxidant capacity (TAC) as well as glucose-6-phosphatase (G6Pase) in liver homogenates and lipid peroxidation (LPO-malondialdehyde-MDA) levels in liver homogenates and liver microsomal fractions against carbon tetrachloride (CCl4)-induced acute liver injury in low-protein fed Sprague-Dawley rats (Rattus norvegicus) were investigated. A group of male Sprague-Dawley rats maintained on a low-protein diet for 5 days were divided into three subgroups. Two subgroups were injected with carbon tetrachloride and the other group with an equivalent amount of olive oil. Two hours after CCl4 intoxication one of the two subgroups was administered with pumpkin seed protein isolate and thereafter switched onto a 20% pumpkin seed protein isolate diet. The other two groups of rats were maintained on the low-protein diet for the duration of the investigation. Groups of rats from the different subgroups were killed at 24, 48 and 72 h after their respective treatments. After 5 days on the low-protein diet the activity levels of all the enzymes as well as antioxidant levels were significantly lower than their counterparts on a normal balanced diet. However, a low-protein diet resulted in significantly increased levels of lipid peroxidation. The CCl4 intoxicated rats responded in a similar way, regarding all the variables investigated, to their counterparts on a low-protein diet. The administration of pumpkin seed protein isolate after CCl4 intoxication resulted in significantly increased levels of all the variables investigated, with the exception of the lipid peroxidation levels which were significantly decreased. From the results of the present study it is concluded that pumpkin seed protein isolate administration was effective in alleviating the detrimental effects associated with protein malnutrition and CCl4 intoxication. It is therefore apparent that pumpkin seed protein isolate has components that have antiperoxidative properties. Copyright © 2006 John Wiley & Sons, Ltd. [source] Effect of pumpkin seed (Cucurbita pepo) protein isolate on the activity levels of certain plasma enzymes in CCl4 -induced liver injury in low-protein fed ratsPHYTOTHERAPY RESEARCH, Issue 4 2005C. Z. Nkosi Abstract The effects of pumpkin seed (Cucurbita pepo) protein isolate on the activity levels of lactate dehydrogenase (LD), alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP) against carbon tetrachloride (CCl4)-induced acute liver injury in low-protein fed rats were investigated. A group of male Sprague-Dawley rats maintained on a low-protein diet for 5 days were divided into three subgroups. Two subgroups were injected with carbon tetrachloride and the other group with an equivalent amount of olive oil. Two hours after CCl4 intoxication one of the two subgroups was administered with pumpkin seed protein isolate. All three subgroups of rats were maintained on the low-protein diet for the duration of the investigation. Groups of rats from the different subgroups were killed at 24, 48 and 72 h after their respective treatments. After 5 days on the low-protein diet the activity levels of all four enzymes were significantly higher than their counterparts on a normal balanced diet. CCl4 intoxication resulted in significant increases in the activity levels of all four enzymes investigated. The administration of pumpkin seed protein isolate after CCl4 intoxication resulted in significantly reduced activity levels of all four enzymes. It is concluded that pumpkin seed protein isolate administration was effective in alleviating the detrimental effects associated with protein malnutrition. Copyright © 2005 John Wiley & Sons, Ltd. [source] Complete Freund's adjuvant immunization prolongs survival in experimental prion disease in miceJOURNAL OF NEUROSCIENCE RESEARCH, Issue 2 2003Yuval Tal Abstract We recently reported that immunization of mice with certain self-prion protein peptides induced specific T-cell and B-cell immune responses; importantly, this immunization was associated with a decrease in the number of protease-resistant PrPSc particles recoverable in a transplanted, scrapie-infected syngeneic tumor. The present study was carried out to determine whether immunization with the immunogenic PrP peptides might influence the natural history of experimental scrapie in mice. We immunized C57BL/6 mice with self-prion peptides in complete Freund's adjuvant (CFA) or with CFA alone as a control and then infected the mice with mouse-adapted scrapie by injection either intraperitoneally or intracerebrally. We report here that immunization with CFA, irrespective of whether prion peptides were present in the inoculum, resulted in marked prolongation of survival of the mice, whether the challenge was intracerebral or intraperitoneal. Mice in the immunized and control groups that died contained equivalent amounts of PrPSc. Thus, CFA immunization has a therapeutic effect in experimental scrapie in mice, possibly by reducing the rate of PrPSc accumulation in the brain. © 2002 Wiley-Liss, Inc. [source] Ethanol Consumption Increases Nitric Oxide Production in Rats, and Its Peroxynitrite-Mediated Toxicity Is Attenuated by PolyenylphosphatidylcholineALCOHOLISM, Issue 6 2002Enrique Baraona Background: Nitric oxide generally mediates beneficial responses but becomes deleterious when coexistence with enhanced superoxide formation leads to the synthesis of peroxynitrite, a potent oxidant and nitrating agent. Methods: To study the effects of ethanol and polyenylphosphatidylcholine on nitric oxide metabolism and toxicity, 36 rats were pair-fed liquid diets with 36% of energy either as ethanol or as additional carbohydrate for 24 days and were killed 90 min after intragastric feeding. Half received polyenylphosphatidylcholine in the diet (3 g/liter), and the other half equivalent amounts of essential fatty acids and choline. Nitric oxide was measured by chemiluminescence in arterial blood and liver cytosol and as a product of the inducible nitric oxide synthase activity. Peroxynitrite formation was assessed by the increase in nitrotyrosine protein residues, measured immunochemically. Results: In blood, administration of ethanol with or without polyenylphosphatidylcholine doubled nitric oxide levels. In the liver, ethanol increased nitric oxide by 52% (p < 0.01), and polyenylphosphatidylcholine attenuated this effect. Ethanol consumption increased the cytosolic activity of the inducible nitric oxide synthase and induced microsomal cytochromes P-450 capable of producing both nitric oxide and superoxide. This was associated with an 18% (p < 0.01) increase in nitrotyrosine protein residues, products of peroxynitrite toxicity, which occurred predominantly in steatotic hepatocytes. Polyenylphosphatidylcholine attenuated these changes by decreasing the ethanol effect on both the cytosolic and the microsomal activities, in addition to acting as a powerful antioxidant. Acute administration of the same ethanol dose increased nitric oxide levels, but did not affect nitrotyrosine protein residues. Conclusions: Chronic, but not acute, ethanol administration increases peroxynitrite hepatotoxicity by enhancing concomitant production of nitric oxide and superoxide, both of which are prevented by polyenylphosphatidylcholine. [source] Aqueous Colloidal Processing of ZTA CompositesJOURNAL OF THE AMERICAN CERAMIC SOCIETY, Issue 1 2009Susana M. Olhero Two different zirconia-alumina composites, ZTA-30 (70 wt% Al2O3+30 wt% ZrO2) and ZTA-60 (40 wt% Al2O3+60 wt% ZrO2), with potential for orthopedic applications, were processed in aqueous media and consolidated by slip casting (SC), hydrolysis-assisted solidification (HAS), and gelcasting (GC) from suspensions containing 50 vol% solids loading. For comparison purposes, the same ceramic compositions were also consolidated by die pressing of freeze-dried granules (FG). In the HAS process, 5 wt% of Al2O3 in the precursor mixture was replaced by equivalent amounts of AlN to promote the consolidation of the suspensions. Ceramics consolidated via GC exhibited higher green (three-point bend) strengths (,17 MPa) than those consolidated by other techniques. Further, these ceramics also exhibited superior fracture toughness and flexural strength properties after sintering for 1 h at 1600°C in comparison with those consolidated by other techniques, including conventional die pressing (FG). [source] Cell wall phenolics of white and green asparagusJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 6 2005Rocío Rodríguez Abstract Cinnamic acids influence physical and textural attributes of plant foods as they play an important role in lignification and cross-linking of cell-wall polymers. Several trans -cinnamic acids have been detected in asparagus cell wall (CW), and, of these, amounts of ferulic acid (FA) and its dehydrodimers have been found to increase significantly during post-harvest storage. The distribution of these compounds along the different sections of white and green spears, as well as their modifications during postharvest storage, has been investigated. It has been observed that the cell walls from the apical, middle and lower sections of the green spears contain equivalent amounts of FA derivatives (about 400 µg g,1 CW), while these compounds are mainly located in the middle (681 µg g,1 CW) and lower section (975 µg g,1 CW) of the white ones. During asparagus post-harvest storage a general increase of FA monomers and dimers took place that affected every section of both green and white spears. Major changes occurred in the middle and lower sections of the white asparagus, where the amounts of ferulic derivatives increased to 1700 and 1678 µg g,1 CW after storage. A similar but less pronounced trend was observed for the green spears. Copyright © 2005 Society of Chemical Industry [source] Characterisation of Chilean hazelnut (Gevuina avellana) tissues: light microscopy and cell wall polysaccharidesJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 3 2003Fernando Dourado Abstract By applying several differential staining techniques and light microscopy, the structure and composition of Chilean hazelnut (Gevuina avellana) seeds were analysed. The structure of the G avellana seed is very simple, with a thin, heavily lignified seed coat and two voluminous cotyledons. The embryo food reserves are uniformly distributed over the cotyledon cells. The cell wall polysaccharides were recovered from the alcohol-insoluble residue by mild treatment with warm chlorite solution and sequential extraction with alkali solutions of increasing concentration. FT-IR spectra in the 1200,850,cm,1 region were used together with chemometric techniques to distinguish the hemicellulosic and pectic polysaccharides in the extracts. The most abundant extracts were fractionated by graded precipitation in ethanol. A xyloglucan was identified by 1H and 13C NMR as the major hemicellulosic polysaccharide, with a sugar composition of 4Glc:3.5Xyl:1Gal:0.5Fuc. The hazelnut cell walls are composed of equivalent amounts of pectic polysaccharides, xyloglucans and cellulose. © 2003 Society of Chemical Industry [source] Baseline Leptin Levels Predict Change in Leptin Levels During Weight Loss in Obese Breast Cancer SurvivorsTHE BREAST JOURNAL, Issue 2 2007Ananda Sen PhD Abstract:, Leptin is an adipocyte-derived hormone involved in regulation of satiety, and it also appears to have a role in breast cancer risk. Leptin therefore might be a useful indicator of the potential preventive effects of weight loss in breast cancer survivors. In this study we examined whether the change in leptin levels could be predicted by weight loss in obese breast cancer survivors. The subjects in this study were participating in a randomized trial of an individualized approach towards weight loss in Detroit, MI. Breast cancer survivors (body mass index of 30,44 kg/m2) were enrolled and fasting blood samples were obtained for leptin analysis over 1 year of study. Leptin levels were available from at least two time points for 36 women, and weight change ranged from a gain of 11% to a loss of 25% of baseline weight. Using a repeated-measures regression model, both baseline leptin level and concurrent percent body fat were found to synergistically predict leptin levels. Thus, for women with the same body fat, those with higher baseline leptin levels are predicted to exhibit smaller decreases in leptin with weight loss. Similar results were obtained for body weight and body weight change, but the associations with body fat were stronger. Breast cancer survivors with initially higher leptin levels may differ with regard to regulation of change in leptin during weight loss resulting in relatively smaller changes in leptin with equivalent amounts of weight loss. [source] Safety of Administration of Human Butyrylcholinesterase and its Conjugates with Soman or VX in RatsBASIC AND CLINICAL PHARMACOLOGY & TOXICOLOGY, Issue 5 2010Raymond F. Genovese Rats were trained on a multiple Fixed-Ratio 32, Extinction 30 sec. (FR32, Ext30) schedule of food reinforcement and then injected (i.m.) with Hu BChE (30 mg/kg), equivalent amounts of Hu BChE,soman conjugate (GDC), Hu BChE,VX conjugate, oxotremorine (OXO) (0.316 mg/kg) or vehicle (n = 8, each group). On the day of injection and on 10 subsequent daily sessions, performance was evaluated on the FR32, Ext30 schedule. Neither conjugates nor Hu BChE produced a performance deficit under the schedule. OXO produced a substantial decrease in responding on the day of administration, with complete recovery observed on subsequent sessions. None of the treatments affected circulating acetylcholinesterase (AChE) activity when evaluated 24,72 hr after injection. The dose of Hu BChE produced a 20,000-fold increase above baseline in circulating BChE activity. Pathological evaluation of organ systems approximately 2 weeks following administration of conjugates or Hu BChE alone did not show toxicity. Taken together, these results suggest that Hu BChE , nerve agent conjugates produced following bioscavenger protection against nerve agents soman and VX do not appear to be particularly toxic. These results add to the safety assessment of Hu BChE as a bioscavenger countermeasure against nerve agent exposure. [source] Simultaneous determination and pharmacokinetic study of oxymatrine and matrine in beagle dog plasma after oral administration of Kushen formula granule, oxymatrine and matrine by LC-MS/MSBIOMEDICAL CHROMATOGRAPHY, Issue 8 2007Yiqi Wang Abstract A rapid, specific and sensitive LC-MS/MS method was developed for the determination of oxymatrine (OMT) and matrine (MT) in beagle dog plasma. The method was applied to study the pharmacokinetics of OMT and MT after oral administration of OMT, MT and Kushen formula granule (KFG) containing equivalent amounts of OMT and MT in a three-period crossover design. The analysis was carried out on an Acquity UPLCÔ BEH C18 column by linear gradient elution with 0.01% acetic acid,water,methanol as mobile phase. Detection was by positive ion electrospray ionization (ESI) mass spectrometry with multiple-reaction monitoring (MRM). Linear calibration curves were both obtained over the concentration range 15,2000 ng/mL, with a limit of quantification of 15 ng/mL. The matrix effect was minimized. The intra- and inter-day precisions (RSDs) were less than 12.4 and 14.7%, respectively, and the accuracy (RE) was from ,2.1 to 2.7%. The validated method was used to determine the concentration,time profiles of OMT and MT. The results indicated that the absorption of OMT and MT after oral administration of KFG was significantly greater than that after oral administration of pure components. Copyright © 2007 John Wiley & Sons, Ltd. [source] Highly efficient deletion of FUT8 in CHO cell lines using zinc-finger nucleases yields cells that produce completely nonfucosylated antibodiesBIOTECHNOLOGY & BIOENGINEERING, Issue 5 2010Laetitia Malphettes Abstract IgG1 antibodies produced in Chinese hamster ovary (CHO) cells are heavily ,1,6-fucosylated, a modification that reduces antibody-dependent cellular cytotoxicity (ADCC) and can inhibit therapeutic antibody function in vivo. Addition of fucose is catalyzed by Fut8, a ,1,6-fucosyltransferase. FUT8,/, CHO cell lines produce completely nonfucosylated antibodies, but the difficulty of recapitulating the knockout in protein-production cell lines has prevented the widespread adoption of FUT8,/, cells as hosts for antibody production. We have created zinc-finger nucleases (ZFNs) that cleave the FUT8 gene in a region encoding the catalytic core of the enzyme, allowing the functional disruption of FUT8 in any CHO cell line. These reagents produce FUT8,/, CHO cells in 3 weeks at a frequency of 5% in the absence of any selection. Alternately, populations of ZFN-treated cells can be directly selected to give FUT8,/, cell pools in as few as 3 days. To demonstrate the utility of this method in bioprocess, FUT8 was disrupted in a CHO cell line used for stable protein production. ZFN-derived FUT8,/, cell lines were as transfectable as wild-type, had similar or better growth profiles, and produced equivalent amounts of antibody during transient transfection. Antibodies made in these lines completely lacked core fucosylation but had an otherwise normal glycosylation pattern. Cell lines stably expressing a model antibody were made from wild-type and ZFN-generated FUT8,/, cells. Clones from both lines had equivalent titer, specific productivity distributions, and integrated viable cell counts. Antibody titer in the best ZFN-generated FUT8,/, cell lines was fourfold higher than in the best-producing clones of FUT8,/, cells made by standard homologous recombination in a different CHO subtype. These data demonstrate the straightforward, ZFN-mediated transfer of the Fut8, phenotype to a production CHO cell line without adverse phenotypic effects. This process will speed the production of highly active, completely nonfucosylated therapeutic antibodies. Biotechnol. Bioeng. 2010;106: 774,783. © 2010 Wiley Periodicals, Inc. [source] In vitro evaluation of bevacizumab toxicity on a retinal ganglion cell lineACTA OPHTHALMOLOGICA, Issue 6 2009Rajesh K. Sharma Abstract. Purpose:, The effects of bevacizumab on cell viability and proliferation in a commonly used retinal ganglion cell line, RGC-5, were examined. Methods:, RGC-5 cells were exposed to 0.1 mg/ml, 1 mg/ml and 2 mg/ml of commercially available bevacizumab in vitro. To examine the specificity of effects, cells were also cultured with increasing and comparable concentrations of proteins (increasing the concentration of proteins in the culture media by 0.1 mg/ml, 1 mg/ml and 2 mg/ml by using additional fetal bovine serum [FBS] and bovine serum albumin [BSA]). Cell proliferation was assessed using a WST-1 kit, crystal violet staining and bromodeoxyuridine (BrdU) incorporation. Cytotoxic effects were assessed by quantifying cell numbers in proliferation-deficient RGC-5 following exposure to bevacizumab using the WST-1 kit, microscopic examination of cells stained with propidium iodide (PI) cells and flow cytometry for differential staining with PI. Results:, Bevacizumab was not toxic to RGC-5 cells in the tested concentrations. It had a stimulatory effect on cell proliferation. A stimulatory effect on proliferation was also noted when equivalent amounts of proteins from FBS or BSA were used, which suggests that bevacizumab may stimulate proliferation non-specifically by increasing the protein contents of the cell growth environment. Conclusions:, Results suggest that intravitreal injection of bevacizumab could alter the internal milieu of the eye by increasing protein concentrations to elicit functional responses in retinotypic cells. This may be especially relevant for cells outwith the control of vascular endothelial growth factor. [source] Modulation of the Aerobic Oxidative Polymerization in Phenylazomethine Dendrimers Assembling Copper ComplexesCHEMISTRY - A EUROPEAN JOURNAL, Issue 36 2010Dr. Takane Imaoka Abstract The aerobic oxidative polymerization of phenol derivatives can provide poly(phenylene oxide)s, which are known as engineering plastics. This oxidation can be carried out with atmospheric oxygen molecules as the oxidizing reagent in the presence of copper complexes as the catalyst; however, stoichiometric or excess amounts of bases are also generally required. By using a phenylazomethine dendrimer complexed with several equivalent amounts of copper chloride, the additive (base)-free polymerization of 2,6-difluorophenol was successful with a very small amount of the catalyst (0.7,mol,% of copper for the monomer) because the dendrimer was composed of many Schiff base units, affording a base and catalyst (copper complex) condensed reaction field. The resulting polymer was nearly linear and the molecular weight was very high. When the equimolar amount of the copper complex in one dendrimer molecule was increased, the polymer obtained under this reaction condition was rather branched, resulting in a higher glass transition temperature. [source] | ||||||||||||||||||||||