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  • Biochemistry and Biotechnology  (526)
  • Cell & Developmental Biology  (419)
  • 2020-2022
  • 1995-1999  (859)
  • 1960-1964  (86)
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  • 1
    Electronic Resource
    Electronic Resource
    CHO DUKX cell lineages preadapted to growth in serum-free suspension culture enable rapid development of cell culture processes for the manufacture of recombinant proteins (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 52 (1996), S. 518-528 
    Details
    ISSN: 0006-3592
    Keywords: CHO cells ; serum-free medium ; adaptation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Using an adaptive strategy, Chinese hamster ovary (CHO) cell lines were developed that are capable of robust growth in serum-free suspension culture. These preadapted derivatives of the commonly used strain of CHO cells (CHO DUKX), termed PA-DUKX, were used for the introduction and stable expression of several heterologous human genes. A significant advantage of recombinant PA-DUKX cells was their ability to readily resume growth in serum-free suspension culture after transfection and amplification of heterologous genes. Expression of recombinant human proteins in PA-DUKX cells was quantitatively similar to that of lineages generated using conventional CHO DUKX cells. In addition, recombinant human proteins expressed by transfected PA-DUKX lineages were shown to be biochemically and structurally similar to those expressed in CHO DUKX cells, PA-DUKX host cell technology provides an opportunity for reducing the time and resources required to develop large-scale, suspension culture-based manufacturing processes employing serum-free medium. © 1996 John Wiley & Sons, Inc.
    Additional Material: 7 Ill.
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  • 2
    Electronic Resource
    Electronic Resource
    Deactivation and conformational changes of cutinase in reverse micelles (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 58 (1998), S. 380-386 
    Details
    ISSN: 0006-3592
    Keywords: reverse micelles ; cutinase ; deactivation ; conformational changes ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Deactivation data and fluorescence intensity changes were used to probe functional and structural stability of cutinase in reverse micelles. A fast deactivation of cutinase in anionic (AOT) reverse micelles occurs due to a reversible denaturation process. The deactivation and denaturation of cutinase is slower in small cationic (CTAB/1-hexanol) reverse micelles and does not occur when the size of the cationic reverse micellar water-pool is larger than cutinase. In both systems, activity loss and denaturation are coupled processes showing the same trend with time. Denaturation is probably caused by the interaction between the enzyme and the surfactant interface of the reversed micelle. When the size of the empty reversed micelle water-pool is smaller than cutinase (at W0 5, with W0 being the water:surfactant concentration ratio) a three-state model describes denaturation and deactivation with an intermediate conformational state existing on the path from native to denaturated cutinase. This intermediate was clearly detected by an increase in activity and shows only minor conformational changes relative to the native state. At W0 20, the size of the empty water-pool was larger than cutinase and the data was well described by a two-state model for both anionic and cationic reverse micelles. For AOT reverse micelles at W0 20, the intermediate state became a transient state and the deactivation and denaturation were described by a two-state model in which only native and denaturated cutinase were present. For CTAB/1-hexanol reverse micelles at W0 20, the native cutinase was in equilibrium with an intermediate state, which did not suffer denaturation. 1-Hexanol showed a stabilizing effect on cutinase in reverse micelles, contributing to the higher stabilities observed in the cationic CTAB/1-hexanol reverse micelles. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:380-386, 1998.
    Additional Material: 2 Ill.
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  • 3
    Electronic Resource
    Electronic Resource
    Comparative study of the colchicine binding site and the assembly of fish and mammalian microtubule proteins (1995)
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 30 (1995), S. 153-163 
    Details
    ISSN: 0886-1544
    Keywords: colchicine binding site ; MTC ; cod microtubules ; bovine microtubules ; MAPs ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Isolated microtubules from cod (Gadus morhua) are apparently more stable to colchicine than bovine microtubules. In order to further characterize this difference, the effect of the colchicine analogue 2-methoxy-5-(2,3,4-trimethoxyphenyl)-2,4,6-cyclo heptatrien-1-one (MTC) was studied on assembly, as measured by turbidity and sedimentation analysis, and on polymer morphology. MTC has the advantage to bind fast and reversible to the colchicine binding site of tubulin even at low temperatures. It was found to bind to one site in cod brain tubulin, with affinity (6.5 ± 1.5) × 105M 1at both low or high temperature, similarly to bovine brain tubulin. However, the effect of the binding differed. At substoichiometric concentrations of MTC bovine brain microtubule assembly was almost completely inhibited, while less effect was seen on the mass of polymerized cod microtubule proteins. A preformed bovine tubulin-colchicine complex inhibited the assembly of both cod and bovine microtubules at substoichiometric concentrations, but the effect on the assembly of cod microtubules was less. At higher concentrations (5 × 10-5 to 1 × 10-3M), MTC induced a large amount of cold-stable spirals of cod proteins, whereas abnormal polymers without any defined structure were formed from bovine proteins. Spirals of cod microtubule proteins were only formed in the presence of microtubule associated proteins (MAPs), indicating that the morphological effect of MTC can be modulated by MAPs. The effects of colchicine and MTC differed. At 10-5M colchicine no spirals were formed, while at 10-4M and 10-3M, a mixture of spirals and aggregates was found. The morphology of the spirals differed both from vinblastine spirals and from the spirals previously found when cod microtubule proteins polymerize in the presence of high Ca2concentrations. The present data show that even if the colchicine binding site is conserved between many different species, the bindings have different effects which seem to depend on intrinsic properties of the different tubulins. © 1995 Wiley-Liss, Inc.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cm.970300207
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  • 4
    Electronic Resource
    Electronic Resource
    Effect of cDNA copy number on secretion rate of activated protein C (1995)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 46 (1995), S. 22-27 
    Details
    ISSN: 0006-3592
    Keywords: cDNA copy number ; gene dosage ; recombinant protein production ; posttranslational modification ; BHK ; secretion ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The secretion rate of activated protein C (APC) by BHK cells was increased 35-fold by increasing the cDNA copy number per cell from 50 to 240. In this range, the relation between APC secretion and cDNA copy number was not linear and the rate of APC secretion per cDNA copy increased sevenfold. This apparent cooperative effect of multiple cDNA copies could be related to their integration in tandem. For cDNA copy numbers higher than 240, the APC secreation rate per cDNA and per cell decreased dramatically. The γ-carboxylation of glutamic acid residues, a posttranslational modification required for APC biological activity, was also investigated. The proportion of APC that was fully γ-carboxylated decreased as the secretion rate of APC increased. © 1995 John Wiley & Sons, Inc.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260460104
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  • 5
    Electronic Resource
    Electronic Resource
    Nitrite inhibition of denitrification by Pseudomonas fluorescens (1995)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 46 (1995), S. 194-201 
    Details
    ISSN: 0006-3592
    Keywords: denitrification kinetics ; nitrite inhibition model ; Pseudomonas fluorescens ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Using a pure culture of Pseudomonas fluorescens as a model system nitrite inhibition of denitrification was studies. A mineral media with acetate and nitrate as sole electron donor and acceptor, respectively, was used. Results obtained in continuous stirred-tank reactors (CSTR) operated at pH values between 6.6 and 7.8 showed that growth inhibition depended only on the nitrite undissociated fraction concentration (nitrous acid). A mathematical model to describe this dependence is put forward. The maximum nitrous acid concentration compatible with cell growth and denitrification activity was found to be 66 μg N/L. Denitrification activity was partially associated with growth, as described by the Luedeking-Piret equation. However, when the freshly inoculated reactor was operated discontinuosly, nitrite accumulation caused growth uncoupling from denitrification activity. The authors suggest that these results can be interpreted considering that (a) nitrous acid acts as a proton uncoupler; and (b) cultures continuoulsy exposed to nitrous acid prevent the uncoupling effect but not the growth inhibition. Examination of the growth dependence on nitrite concentration at pH 7.0 showed that adapted cultures (grown on CSTR) are less sensitive to nitrous acid inhibition than the ones cultivated in batch. © 1995 John Wiley & Sons, Inc.
    Additional Material: 12 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260460303
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  • 6
    Electronic Resource
    Electronic Resource
    Targeted antisense modulation of inflammatory cytokine receptors (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 55 (1997), S. 72-81 
    Details
    ISSN: 0006-3592
    Keywords: antisense ; receptor targeted delivery ; acute phase response ; IL-6 ; gp 130 ; HepG2 cells ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Antisense technology is potentially a powerful means by which to selectively control gene expression. We have used antisense oligonucleotides to modulate the response of the hepatoma cell line, HepG2, to the inflammatory cytokine, IL-6, by inhibiting the expression of its multifunctional signal transducer, gp130. HepG2 cells respond to IL-6 by upregulating acute phase proteins, such as haptoglobin, by five- to tenfold. Gp130 is central to this response, as the upregulation of haptoglobin is almost completely blocked by the addition of high concentrations (∼100 μg/ml) of a monoclonal antibody to gp 130. Antisense oligodeoxynucleotides complementary to the mRNA encoding gp 130 inhibited the upregulation of haptoglobin by IL-6-stimulated HepG2 cells by about 50%. However, a nonsense sequence also inhibited haptoglobin secretion by about 20%. To improve the specificity and efficiency of action, we targeted the antisense oligonucleotides to HepG2 cells using a conjugate of asialoglycoprotein-poly-L-lysine. The targeted antisense reduced the binding of IL-6 to HepG2 cells, virtually eliminating high affinity binding. In addition, it inhibited haptoglobin upregulation by over 70%. Furthermore, the dose of targeted antisense required for biological effect was reduced by about an order of magnitude as compared with unconjugated antisense. These results demonstrate the potential of antisense oligonucleotides as a means to control the acute phase response as well as the need for a greater understanding of the mechanism and dynamics of antisense molecules as they are developed toward therapeutic application. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55: 72-81, 1997.
    Additional Material: 7 Ill.
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  • 7
    Electronic Resource
    Electronic Resource
    Kinetic modeling of poly(β-hydroxybutyrate) production and consumption by Paracoccus pantotrophus under dynamic substrate supply (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 55 (1997), S. 773-782 
    Details
    ISSN: 0006-3592
    Keywords: PHB ; poly(β-hydroxybutyrate) ; Paracoccus pantotrophus ; dynamic growth ; metabolic modeling ; polymers ; activated sludge process ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The objective of the research was to obtain insights into the behavior of microorganisms under feast/famine conditions as often occur in wastewater treatment processes. The response of microorganisms to such conditions is the accumulation of storage polymers like poly(β-hydroxybutyrate). The research was performed using a pure culture of Paracoccus pantotrophus LMD 94.21. A steady-state C-limited chemostat culture was switched to batch mode and a pulse of acetate was added. As long as external substrate (acetic acid) was present, the organism grew and accumulated poly(β-hydroxybutyrate). After depletion of the external substrate, the stored poly(β-hydroxybutyrate) was used as growth substrate. Poly(β-hydroxybutyrate) accumulation was found to be strongly dependent on the growth rate of the organism before the pulse addition of acetate. Poly(β-hydroxybutyrate) accumulation was correlated to the difference in maximum acetate uptake rate and the acetate required for growth. Based on the interpretation of the experimental results, a metabolically structured model has been set up. This model adequately describes the observed kinetics of the poly(β-hydroxybutyrate) formation and consumption. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55: 773-782, 1997.
    Additional Material: 7 Ill.
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  • 8
    Electronic Resource
    Electronic Resource
    Order-disorder in T, T′, and T* phase: Superconductors and related materials (1995)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 30 (1995), S. 193-207 
    Details
    ISSN: 1059-910X
    Keywords: Superconductivity ; n-type superconductors ; Nonstoichiometry ; Superconducting oxides ; T, T′ ; T* structures ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: After reviewing microstructural studies on superconducting materials showing T, T′, and T* structural types, results are presented on the microstructure of some n-type superconductors and related materials prepared with accurate control of the oxygen stoichiometry. Electron microscopy is used to describe the ordering of interstitial oxygen defects in T-type La2NiO4+δ leading to the formation of the n = 2 term of a homologous series with the general formula La8nNi4nO16n+1. Structural transitions and superstructure formation in the Pr2-x-yCexSryCuO4-δ system are reported, where T, T′:;, and T* phases are isolated as a function of both Ce and Sr content. © 1995 Wiley-Liss, Inc.
    Additional Material: 15 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070300302
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  • 9
    Electronic Resource
    Electronic Resource
    Blood plasma levels of lipoperoxides, glutathione peroxidase, beta carotene, vitamin A and E in women with habitual abortion (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Cell Biochemistry and Function 16 (1998), S. 227-231 
    Details
    ISSN: 0263-6484
    Keywords: lipid peroxidation ; antioxidants ; habitual abortion ; vitamin A and E ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The plasma levels of lipoperoxides, glutathione peroxidase (GSH-Px), reduced glutathione (GSH), beta carotene, vitamin A, E, some plasma biochemical and blood haematological parameters were investigated in 40 women with habitual abortion (HA) and controls. The levels of GSH, vitamin A, E and beta carotene were significantly lower in women with HA than in controls. However, the plasma levels of lipid peroxidation, alkaline phosphatase (ALP), glucose and blood haemoglobin were significantly higher in HA than in controls. In addition, plasma levels of GSH-Px, AST, ALT, total bilirubin, total protein, albumin, sodium, potassium, calcium and number of white blood cells, red blood cells, platelet and values of packet cell volume showed no significant differences between HA and controls. According to the results of this study, we observed that the levels of lipid peroxidation were increased and plasma levels of vitamin A, E and beta carotene were decreased in HA. The decrease of those antioxidants may play a significant role in women with habitual abortion. Copyright © 1998 John Wiley & Sons, Ltd.
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  • 10
    Electronic Resource
    Electronic Resource
    Occurrence of ampulla in the ductus deferens of the Indian garden lizard Calotes versicolor daudin (1995)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 225 (1995), S. 261-268 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: During the breeding season, the terminal end of the ductus deferens of Calotes versicolor appears swollen and is comparable to the ampulla of the mammalian ductus deferens. Its anatomy was studied from paraffin sections. It differentiates along its length into five zones. The first has thick smooth muscle and pesudostratified epithelium; the second has luminal trabeculae with an epithelium showing evidence of secretory activity; the third has the epithelial mucosa abutting against the smooth muscle in the form of pocketlike indentations; the fourth has crypts between epithelial folds; and the fifth zone is a sphincter. The anatomy of this ampullary region is indicative of secretory as well as spermatophagous roles. It undergoes seasonal change and appears to be androgen-dependent. © 1995 Wiley-Liss, Inc.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jmor.1052250303
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