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  • Process Engineering, Biotechnology, Nutrition Technology  (11,711)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Biomedical microdevices 2 (2000), S. 255-264 
    ISSN: 1572-8781
    Keywords: microstamping ; gradients ; electrophoresis ; bio- molecule ; cell culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Technology
    Notes: Abstract We have developed a technique for producing patterned biomolecule gradients on glass substrates using microstamping. The technique uses a microfabricated polydimethylsiloxane (PDMS) stamp with micrometer length features to transfer biomolecules to silane-derivatized substrates. The soaking solution for the stamp is subjected to an electric field to electrophoretically produce a gradient in solution and hence on the stamp. To test the efficacy of the procedure we tested substrates stamped with poly-D-lysine by fluorescently labeling the amine groups and measuring the fluorescence. It is possible to modify the steepness of the gradient using both voltage and electrode spacing, and the profile can be modeled using an isoelectric focusing model. Since gradients have been implicated in roles in guidance and patterning of neurons and other cell types, a flexible method to produce biomolecule gradients using microstamping may further extend the usefulness of microstamping to studies of cell patterning and guidance.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Cytotechnology 19 (1995), S. 95-105 
    ISSN: 1573-0778
    Keywords: antibiotics ; cell culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract This article describes the historical development of the prophylactic use of antibiotics in cell culture as well as their effects on cells. The influence of antibiotics on cell morphology, cellular degeneration and cell death and cellular function is summarized. Cellular DNA as well as protein synthesis are affected which can lead to interference with, or even changes in, metabolic processes. Such effects must be considered in cell culture research. As antibiotics are used in multifold ways, the otherwise standardized conditions in cell culture are no longer comparable. The prophylactic use of antibiotics is rejected for scientific reasons.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Cytotechnology 2 (1989), S. 181-185 
    ISSN: 1573-0778
    Keywords: culture ; media ; skin ; hypertrophic scar ; cell culture ; electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Twenty-five human or mouse tissue samples, some up to 8×4×2 cm, were immersed in a special transport medium (TM), L15M15, up to 7 days before being processed or placed in tissue culture. To test the efficacy of this medium, we concurrently placed pieces of the same tissues in a sterile phosphate buffered solution (PBS). We also tested the preservative capabilities of TM and PBS at room temperature and with refrigeration. Differences between TM and PBS are demonstrated, which are more pronounced using room temperature up to 4 days time. The tissues stored in TM show fewer degenerative or autolytic changes than the same tissue stored in PBS under identical conditions. Using regrigeration further enhanced the preservative qualities of TM up to 4 days, but not PBS. There were no obvious differences between tissues stored in TM and PBS with refrigeration after 7 days. We conclude that transport medium L15M15 is a useful medium for preserving tissue viability, especially large tissue samples, up to 4 days, especially if refrigerated.
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  • 4
    ISSN: 1573-0778
    Keywords: anticancer drugs ; at-MDR ; cell culture ; DNA topoisomerase II ; drug resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The characteristic feature of multidrug resistance (MDR) associated with drugs that interact with DNA topoisomerase II (topo II) is alterations in topo II activity or amount (at-MDR). We have characterized the at-MDR phenotype in human leukemic CEM cells selected for resistance to the topo II inhibitor, VM-26. Compared to drug-sensitive cells, the key findings are that at-MDR cells exhibit (i) decreased topo II activity; (ii) decreased drug sensitivity, activity and amount of nuclear matrix topo II; (iii) increased ATP requirement of topo II; (iv) a single base mutation in topo II resulting in a change of Arg to Gln at position 449, at the start of the motif B/nucleotide binding site; and (v) decreased topo II phosphorylation, suggesting decreased kinase or increased phosphatase activities. Recent results using single-stranded conformational polymorphism analysis reveals the presence of a mutation in the motif B/nucleotide binding site of the topo IIα gene in CEM at-MDR cells and in another leukemic cell line selected for resistance to m-AMSA. Finally, we have observed marked changes in the nuclear distribution of topo II in cells treated with anti-topo II drugs and have also found these changes to be attenuated in drug-resistant cells. We postulate that traditional inhibitors of topo II alter the equilibrium of the strand-passing reaction such that the number of enzyme-DNA covalent complexes increases. We further suggest that when the enzyme is bound to DNA it is protected from proteolysis, thus allowing more topo II molecules to be detected. We propose that MDR associated with alterations in topo II may have clinical consequences, and our current efforts involve exploiting these biochemical and molecular observations in the development of probes that may be useful to identify such drug resistant cells in the tumors of patients.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Cytotechnology 11 (1993), S. 169-174 
    ISSN: 1573-0778
    Keywords: cell culture ; hybridoma ; monoclonal antibody ; serum-free medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The effects of several different substances, including insulin, transferrin, ethanolamine, selenite and butyrate on the growth of murine hybridoma 2F7 cells, which secrete monoclonal antibody against small cell lung cancer, were investigated, and a serum-free medium SFMI was formulated. The effects of taurine, spermidine, progesterone and adenine on the cell growth were tested further on the basis of the medium SFMI, and a modified serum-free medium SFM II was established. On the basis of medium SFM II, the substitution tests of ferric citrate for transferrin were carried out, and it was found that transferrin could be replaced. The experiments suggested that the formulated serum-free medium was suitable for 2F7 cell growth and monoclonal antibody secretion, and thus facilitated subsequent purification of monoclonal antibody.
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  • 6
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    Springer
    Cytotechnology 11 (1993), S. 219-231 
    ISSN: 1573-0778
    Keywords: cell culture ; chelators ; metal ion buffer ; serum-free medium ; serum replacement (serum substitute) ; trace elements
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The development of a new stable synthetic serum replacement (SSR) is described, which allows the cultivation of mammalian cells in a defined, protein-free medium containing only dialyzable components. With a low concentration of insulin (RPMI-SR2 medium), growth rates of the transformed cell lines L929, HELA S3, and the hybridoma 1E6 were comparable to growth rates obtained with a serum-containing medium. The same medium also supported long-term cultivation of non-dividing mouse macrophages. The main principle of SSR is a metal ion buffer containing a balanced mixture of iron and trace metals. Stability against precipitation of important metals is achieved by the combined use of EDTA and citric acid as chelating agents. Efficient iron supply is mediated through the inclusion of the compound Aurintricarboxylic acid as a synthetic replacement for transferrin. SSR also contains a growth-promoting surfactant, Pluronic F68. Thus SSR provides a general foundation for growth and differentiation normally provided by serum. Limitations of other serum-free medium designs are discussed here: 1) the inability of transferrin to chelate all metals in the medium; and 2) the use of inorganic iron salts or iron citrate as an iron supplement leads to rapid precipitation of iron hydroxide in the medium. Both these problems are solved in the design of SSR.
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  • 7
    ISSN: 1573-0778
    Keywords: cell culture ; porous microcarrier ; prourokinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A recombinant DNA CHO cell line secretingurokinase-type plasminogen activator (u-PA) wascultivated with Cytopore cellulose porousmicrocarriers in a 30l Biostat UC stirred tankreactor. After 26 days of culture, using a spinfilter toretain cells in bioreactor, the cell density couldreach 1.33 × 107 ml-1. The maximal u-PAactivity in supernatant was 7335 IU·ml-1, and204l supernatant containing 7.1 g u-PA was harvested.After 100 days of culture with 0.1% fetal bovineserum medium, a modified cell retention system whichcan be washed-out backward, substituted thespinfilter to prevent filter clogging. The maximalcell density was over 107 ml-1, the maximalu-PA activity in supernatant reached 6250IU·ml-1, and 1604l supernatant containing about51 g u-PA was harvested. Compared to perfusionculture, batch medium-replaced culture could raiseutilizing efficiency of the medium, increase cell specificproductivity and improve the quality of the product which wasnot steady in a 37 °C environment. Cells can movefrom seed porous microcarriers occupied by cells tovacant microcarriers spontaneously, withouttrypsinization, and continue to grow until all microcarriers contained cells. It shows that Cytoporeporous microcarriers are very useful and convenient toscale up cultivation step by step.
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  • 8
    ISSN: 1573-0778
    Keywords: cell culture ; flow injection analysis ; glycosylation ; macro-heterogeneity ; prolactin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A flow injection analysis (FIA) system is presented for a twostep immunoassay-based determination of the total humanprolactin (hPRL) concentration along with its degree ofglycosylation. Separate measurement of total hPRL and nonglysosylated human prolactin (nG-hPRL) were made using twoflow-through cartridges each containing immobilized antibodiesof different specificity. The antibodies are immobilized on thesurface of a carrier. Glycosylated hPRL (G-hPRL) and, thus, thedegree of glycosylation were calculated by the differencebetween the two specific determinations. Enhanced specificityfor the determination of nG-hPRL was obtained using unfavorablebinding conditions through incorporation of alkaline pH andchaotropic agents into the carrier/dispersion buffer. The assayfor total hPRL and nG-hPRL were each found to be linear withinthe relevant concentration range. The results of the two-stepFIA method were found to agree with those obtained by thestandard methods of ELISA and western blotting while offeringthe advantage of minimal analysis time (10 min) and eliminationof manual manipulations.
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  • 9
    ISSN: 1573-0778
    Keywords: cell culture ; half-life ; packaging cells ; retrovirus ; titer ; ΨCRIP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Rapidly expanding development and practice of gene therapy requires the availability of large quantities of high titer retroviral supernatants. One way to achieve high retroviral titers is through improved understanding of the kinetics of retroviral production and decay, and the subsequent development of improved cell culture methods. In the present study we investigated the effects of different operational modes on the retroviral production of the NIH 3T3 fibroblast derived amphotropic murine retroviral producing cell line pMFG/ΨCRIP. Semi-continuous culture (exchange of 50% of medium volume daily) was found to promote cell growth and enhance retroviral production. The rapid medium exchange resulted in significantly larger amounts of high titer supernatants and an extended production phase as compared to the batch control cultures. The specific viral productivity of the pMFG/ΨCRIP cells was in the range of 10 to 40 infectious viruses produced per thousand producer cells per day. The CV-1 African Green Monkey kidney cell line was used as the infection target. Lowering the serum level form 20% to 10% improved retroviral production slightly. However, at lower serum levels (1%, 5% and 10% (v/v)) growth of the producer cell line, and thus retroviral production, was directly proportional to the serum level. The half-life of the virus at 37°C was found to be 5.5 hours. Promoting the growth of producer cell lines can improve retroviral vectors titers and viral production. High cell density systems that allow for rapid cell growth and waste product removal are likely to be used to generate high-titer retroviral supernatants.
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  • 10
    ISSN: 1573-0778
    Keywords: cell culture ; lymphocyte ; lymphokine-activated killer cell ; recombinant interleukin 2 ; serum-free medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract We examined a serum-free medium (designated as TYI 101) for the generation of lymphokine-activated killer (LAK) cells from human lymphocytes, regional lymph node lymphocytes (RLNL) and peripheral blood lymphocytes (PBL). TYI 101 medium consisted of, in addition to nutrient mixture, transferrin, insulin, fetuin, sodium selenite, 2-mercaptoethanol, o-phosphorylethanolamine, chick egg yolk and porcine kidney extract. These hormones were effective for supporting RLNL proliferation as assessed by (3H)-thymidine uptake. When human lymphocytes from two different sources were cultivated with recombinant interleukin 2 (rIL-2) in TYI 101 medium, LAK activity was generated. In cultures of PBL from a healthy donor, LAK cells were generated in TYI 101 medium as efficiently as in RPMI 1640 medium supplemented with 10% human AB-type serum (RPMI-AB). In cultures of RLNL from lung cancer patients, LAK activity obtained in TYI 101 medium was about sixty-five percent of that in RPMI-AB. However, the addition of a small amount of AB-type serum improved the generation of LAK activity, LAK cell expansion, and cell viability in TYI 101 medium. We conclude that TYI 101 medium can be used for the generation of LAK cells from human lymph node lymphocytes with supplementation of none or only a reduced amount of human serum.
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  • 11
    ISSN: 1573-0778
    Keywords: adoptive immunotherapy ; cell culture ; cell culture apparatus ; Interleukin-2 ; lymphokine-activated killer cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract We recently developed a new culture system based on dialysis perfusion (designated JCC-device) for the generation and expansion of human lymphokine-activated killer (LAK) cells (Murata et al., 1990). More recently we have scaled up the volume of the culture vessel of the JCC-device from 100 ml to 400 ml for clinical use. In the present study, using this new 400 ml JCC-device, we cultured human lymph node lymphocytes (LNL) obtained from 8 surgical patients with primary lung cancer, and investigated the cellular characteristics in comparison with a conventional batchwise culture system using tissue culture dishes. With the JCC-device, the cell density reached a maximum 2.7×107 cells/ml with greater than 90% viability by the appropriate exchange of perfusion medium and by making additions at the appropriate intervals for recombinant interleukin-2 (rIL-2). The expansion fold of LNL with the JCC-device, ranging 6.6- to 19.2-fold (mean 13.8-fold), was not significantly different from that in dish cultures. There was no marked difference in cell surface phenotypes between the two culture systems in 7 out of 8 cases. As for LAK activity of LNL, the JCC culture was either superior or equal in 4 out of 8 cases, but inferior in the other 4 cases to the conventional dish cultures. In the latter cases, the usage of serum for the JCC culture was limited, which might have resulted in the low LAK activity. The JCC-device was able to reduce the consumption of basal medium, rIL-2 and serum by 20%, 84% and 96%, respectively compared to the conventional tissue culture systems. The JCC-device improved the routine performance of adoptive immunotherapy with LAK cells and rIL-2.
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  • 12
    ISSN: 1573-0778
    Keywords: cell culture ; enhanced production ; fibroblast cells ; interferon-β ; Langmuir-Blodgett film
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Normal human skin (NB1-RGB) cells were cultured in the presenceof polyinosinic and polycytidylic acids, diethylaminoethyldextran, cycloheximide and actinomycin D, which induced humaninterferon-β. The simplest induction method, that requiredonly polyinosinic and polycytidylic acids and diethylaminoethyldextran was found to give the highest production ofinterferon-β by the cells. The cell growth and productionof interferon-β were investigated for NB1-RGB cellscultured on silk fibroin, poly(γ-methyl-L-glutamate),poly(γ-benzyl-L-glutamate) and collagen films prepared bythe Langmuir-Blodgett (LB) and casting methods. The cell densityof NB1-RGB cells cultured on the LB films was found to be higherthan that on the cast films made of the same polymer. Thisindicates that not only the chemical structure of the polymersused for the preparation of the films but the preparationmethods of the films, i.e., casting and LB methods, are also astrong factor affecting the cell growth. The production ofinterferon-β per unit number of cells was found to behigher on the cast films than that on the LB films made of thesame polymer. This is explained by the fact that the optimalsuppressed growth of NB1-RGB cells on the cast films leads tothe enhanced production of interferon-β on the cast filmscompared to those on the LB films prepared by the same polymer.
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  • 13
    ISSN: 1573-0778
    Keywords: cell culture ; FGF ; growth factors ; protein sequencing ; zwitterionic detergent
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Among several detergents, a zwitterionic detergent, 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS), was found to be least cytotoxic for cultured mammalian cells. CHAPS improved the activity recovery and elution profile of crude and purified fibroblast growth factors (FGFs) during chromatographies. Diluted preparations of FGFs were stabilized by CHAPS against the loss during storage. Amino acid sequence analysis was not disturbed by CHAPS. CHAPS was removable by reversed-phase high-performance liquid chromatography. These results indicate that CHAPS is useful as a non-cytotoxic stabilizing agent in purification of various kinds of bioactive polypeptides.
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  • 14
    ISSN: 1573-0778
    Keywords: hybridoma ; cell volume ; cell culture ; flow cytometry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Light scattering properties of hybridoma cells were examined with flow cytometry. Viable and dead cells form two distinct populations. The distribution of the two populations changes during a batch culture. the concentration of dead cells measured by flow cytometry correlates well to that measured by hemacytometer. The distribution based on small-angle light scattering is similar to the distribution based on volume as measured by Elzone particle counter. It thus appears that viable cells form the population with a larger mean cell volume. The results also indicate that the volume of viable cells decreases during the cultivation while that of dead cells remains relatively constant.
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  • 15
    ISSN: 1573-0778
    Keywords: cell culture ; mixing time ; oxygen demand ; oxygen transfer ; pH and dO2 sensitivity ; scale-up
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Because of concern for cell damage, very low agitation energy inputs have been used in industrial animal cell bioreactors, typical values being two orders of magnitude less than those found in bacterial fermentations. Aeration rates are also very small. As a result, such bioreactors might be both poorly mixed and also unable to provide the higher oxygen up-take rates demanded by more intensive operation. This paper reports experimental studies both of K L a and of mixing (via pH measurements) in bioreactors up to 8 m3 at Wellcome and of scaled down models of such reactors at Birmingham. Alongside these physical measurements, sensitivity of certain cell lines to continuously controlled dO2 has been studied and the oxygen up-take rates measured in representative growth conditions. An analysis of characteristic times and mixing theory, together with other recent work showing that more vigorous agitation and aeration can be used especially in the presence of Pluronic F-68, indicates ways of improving their performance. pH gradients offer a special challenge.
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  • 16
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    Cytotechnology 22 (1996), S. 3-16 
    ISSN: 1573-0778
    Keywords: cell culture ; process monitoring ; oxygenation ; CO2 transfer ; aggregation ; segregation ; diffusion, on-line monitoring
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract High density cell culture systems offer the advantage of production of bio-pharmaceuticals in compact bioreactors with high volumetric production rates; however, these systems are difficult to design and operate. First of all, the cells have to be retained in the bioreactor by physical means during perfusion. The design of the cell retention is the key to performance of high density cell culture systems. Oxygenation and media design are also important for maximizing the cell number. In high density perfusion reactors, variable cell density, and hence the metabolic demand, require constant adjustment of perfusion rates. The use of cell specific perfusion rate (CSPR) control provides a constant environment to the cells resulting in consistent production. On-line measurement of cell density and metabolic activities can be used for the estimation of cell densities and the control of CSPR. Issues related to mass transfer and mixing become more important at high cell densities. Due to the difference in mass transfer coefficients for oxygen and CO2, a significant accumulation of dissolved CO2 is experienced with silicone tubing aeration. Also, mixing is observed to decrease at high densities. Base addition, if not properly done, could result in localized cell lysis and poor culture performance. Non-uniform mixing in reactors promotes the heterogeneity of the culture. Cell aggregation results in segregation of the cells within different mixing zones. This paper discusses these issues and makes recommendations for further development of high density cell culture bioreactors.
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  • 17
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    Cytotechnology 31 (1999), S. 3-8 
    ISSN: 1573-0778
    Keywords: cell culture ; livestock ; milk ; nuclear transfer ; transgenic
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Transgenesis may allow the generation of farm animals with altered phenotype, animal models for research and animal bioreactors. Although such animals have been produced, the time and expense involved in generating transgenic livestock and then evaluating the transgene expression pattern is very restrictive. If questions about the ability and efficiency of expression could be asked solely in vitro rapid progress could be achieved. Unfortunately, experiments addressing transcriptional control in vitro have proved unreliable in their ability to indicate whether a transgene will be transcribed or not. However, initial studies suggest that cell culture may be able to predict in vivo post-transcriptional events. We review these issues and propose that strategies which engineer the transgene integration site could enhance the probability for efficient expression. This approach has now become feasible with the development of techniques allowing animals to be generated from somatic cells by nuclear transfer. The important step in this procedure is the use of cells grown in culture as the source of genetic information, allowing the selection of specific transgene integration events. This technology which has dramatically increased the potential use of transgenic livestock for both agricultural and biotechnological applications, is based on standard cell culture methodology. We are now at the start of a new era in large animal transgenics.
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  • 18
    ISSN: 1573-0778
    Keywords: heartwater ; cell culture ; Cowdria ruminantium ; bovine vascular endothelial cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The causal agent of heartwater disease of domestic ruminants,Cowdria ruminantium, can, with difficulty, be isolated and passaged in lines of bovine endothelial cells grown in the presence of the Glasgow modification of Eagle's minimal essential medium. However, when Leibovitz's L-15 medium supplemented with 0.45% glucose at pH 6.0–6.5 is used as maintenance medium for these cells, isolation and serial passage may routinely be achieved.
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  • 19
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    Cytotechnology 15 (1994), S. 87-94 
    ISSN: 1573-0778
    Keywords: Adaptation ; ammonia ; cell culture ; glutamine ; glutamate ; dipeptides
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Although glutamine is used as a major substrate for the growth of mammalian cells in culture, it suffers from some disadvantages. Glutamine is deaminated through storage or by cellular metabolism, leading to the formation of ammonia which can result in growth inhibition. Non-ammoniagenic alternatives to glutamine have been investigated in an attempt to develop strategies for obtaining improved cell yields for ammonia sensitive cell lines. Glutamate is a suitable substitute for glutamine in some culture systems. A period of adaptation to glutamate is required during which the activity of glutamine synthetase and the rate of transport of glutamate both increase. The cell yield increases when the ammonia accumulation is decreased following culture supplementation with glutamate rather than glutamine. However some cell lines fail to adapt to growth in glutamate and this may be due to a low efficiency transport system. The glutamine-based dipeptides, ala-gln and gly-gln can substitute for glutamine in cultures of antibody-secreting hybridomas. The accumulation of ammonia in these cultures is less and cell yields in dipeptide-based media may be improved compared to glutamine-based controls. In murine hybridomas, a higher concentration of gly-gln is required to obtain comparable cell growth to ala-gln or gln-based cultures. This is attributed to a requirement for dipeptide hydrolysis catalyzed by an enzyme with higher affinity for ala-gln than gly-gln.
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  • 20
    ISSN: 1573-0778
    Keywords: Baculovirus ; cell culture ; Drosophila ; gene expression ; insect cell ; metallothionein promoter ; recombinant protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract In this report, we compare two different expression systems: baculovirus/Sf9 and stable recombinantDrosophila Schneider 2 (S2) cell lines. The construction of a recombinant S2 cell line is simple and quick, and in batch fermentations the cells have a doubling time of 20 hours until reaching a plateau density of 20 million cells/ml. Protein expression is driven by theDrosophila Metallothionein promoter which is tightly regulated. When expressed in S2 cells, the extracellular domain of human VCAM, an adhesion molecule, is indistinguishable from the same protein produced by baculovirus-infected Sf9 cells. Additionally, we present data on the expression of a seven trans-membrane protein, the dopamine D4 receptor, which has been successfully expressed in both systems. The receptor integrates correctly in the S2 membrane, binds [3H]spiperone with high affinity and exhibits pharmacological characteristics identical to that of the receptor expressed in Sf9 and mammalian cells. The general implications for large scale production of recombinant proteins are discussed.
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  • 21
    ISSN: 1573-0778
    Keywords: Mycoplasma ; cell culture ; clinical testing ; microbial screening ; PCR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The polymerase chain reaction (PCR) has been used for the general detection ofMollicutes. 25Mycoplasma andAcholeplasma species were detected including important contaminants of cell cultures such asM. orale, M. arginini, M. hyorhinis, M. fermentans, A. laidlawii and additional human and animal mycoplasmas. PCR reactions were performed using a set of nested primers defined from conserved regions of the 16S rRNA gene. The detection limit was determined to be 1 fg mycoplasma DNA, which is equivalent to 1–2 genome copies of the 16S rRNA coding region. The identity of the amplification products was confirmed by agarose gel electrophoresis and restriction enzyme analysis. DNA from closely and distantly related micro-organisms did not give rise to specific amplification products. The method presented here offers a much more sensitive, specific and rapid assay for the detection of mycoplasmas than the existing ones.
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  • 22
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    Cytotechnology 16 (1994), S. 147-150 
    ISSN: 1573-0778
    Keywords: Hybridoma ; peptone ; monoclonal antibody ; cell culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Hybridoma WuT3 secreting a monoclonal antibody against T lymphocytes was grown in RPMI 1640 medium supplemented with 1% human serum. The effect of the concentration of peptone, as an additive, was investigated on cell growth, monoclonal antibody formation, and cell metabolism over 0–10 g l−1 range. It was found that 1–5 g l−1 peptone can significantly promote the growth of cells and increase the formation of monoclonal antibody, especially at 3–5 g l−1, when both the accumulating level and secretion rate of monoclonal antibody are higher than that at other peptone concentrations. Based on glucose, lactate and ammonia analysis data, the efficiency of glycolysis was assessed and the utilization of amino acids was more efficient at 3–5 g l−1 peptone. The cell growth and monoclonal antibody formation were inhibited at higher peptone concentrations, e.g. 10 g l−1.
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  • 23
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    Cytotechnology 7 (1991), S. 121-130 
    ISSN: 1573-0778
    Keywords: cell culture ; cellulose sulphate ; encapsulation ; monoclonal antibodies ; poly-dimethyl-diallyl-ammoniumchloride
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A new encapsulation method was developed for the cultivation of mammalian cells. The capsules were produced using a solution of sodium cellulose sulphate (CS)(1.5%) and poly-dimethyl-diallyl-ammoniumchloride (PDMDAAC). When CS droplets fell into the precipitation bath consisting of a 2% solution of PDMDAAC, immediately a membrane at the interphase was built up. The influences of varying encapsulation process parameters on capsule characteristics, cell growth, and monoclonal antibody production were tested. This new method showed advantages when compared to other methods mainly due to time simplicity of the whole process.
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  • 24
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    Cytotechnology 4 (1990), S. 181-189 
    ISSN: 1573-0778
    Keywords: cell culture ; microperfusion ; luminal-antiluminal media gradients ; specific support ; extracellular matrix proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The lack of a suitable system to culture epithelial cells for a long period under a luminal-antiluminal medium gradient, was the reason to develop a new system. It consists of an interchangeable sheet of permeable support material, which is set in place by two tight fitting holding rings. For special demands the supports can be coated with extracellular matrix proteins improving cellular attachment and terminal differentiation. The handling of the sheets by forceps proceeds easily and quickly, thus fastening the transfer of cultured cells without additional manipulations. The sheets can be transferred to a newly developed microperfusion chamber on which an apical and a basal perfusion over a long culture period parallel to a transepithelial electrophysiological registration becomes possible. The chamber has an extremely low amount of fluid dead space. The separate perfusion of cultured cells under isotonic, hypotonic or hypertonic conditions opens new possibilities. Thus, culture can be performed under most natural conditions e.g., that found within the kidney.
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  • 25
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    Cytotechnology 5 (1991), S. 31-46 
    ISSN: 1573-0778
    Keywords: serum-free ; cell culture ; carcinoma
    Source: Springer Online Journal Archives 1860-2000
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  • 26
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    Biotechnology and Bioengineering 51 (1996), S. 399-409 
    ISSN: 0006-3592
    Keywords: cell damage ; cell culture ; bubble aeration ; agitation ; bubble coalescence and breakup ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: It has been established that the forces resulting from bubbles rupturing at the free air (gas)/liquid surface injure animal cells in agitated and/or sparged bioreactors. Although it has been suggested that bubble coalescence and breakup within agitated and sparged bioreactors (i.e., away from the free liquid surface) can be a source of cell injury as well, the evidence has been indirect. We have carried out experiments to examine this issue. The free air/liquid surface in a sparged and agitated bioractor was eliminated by completely filling the 2-L reactor and allowing sparged bubbles to escape through an outlet tube. Two identical bioreactors were run in parallel to make comparisons between cultures that were oxygenated via direct air sparging and the control culture in which silicone tubing was used for bubble-free oxygenation. Thus, cell damage from cell-to-bubble interactions due to processes (bubble coalescence and breakup) occurring in the bulk liquid could be isolated by eliminating damage due to bubbles rupturing at the free air/liquid surface of the bioreactor. We found that Chinese hamster ovary (CHO) cells grown in medium that does not contain shear-protecting additives can be agitated at rates up to 600 rpm without being damaged extensively by cell-to bubble interactions in the bulk of the bioreactor. We verified this using both batch and high-density perfusion cultures. We tested two impeller designs (pitched blade and Rushton) and found them not to affect cell damage under similar operational conditions. Sparger location (above vs. below the impeller) had no effect on cell damage at higher agitation rates but may affect the injury process at lower agitation intensities (here, below 250 rpm). In the absence of a headspace, we found less cell damage at higher agitation intensities (400 and 600 rpm), and we suggest that this nonintuitive finding derives from the important effect of bubble size and foam stability on the cell damage process. © 1996 John Wiley & Sons, Inc.
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  • 27
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    Cytotechnology 2 (1989), S. 135-140 
    ISSN: 1573-0778
    Keywords: hepatoma ; cell culture ; microcarrier ; agitation rate ; cell inoculation density ; growth rate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Hepatoma cells, HepG2, grew normally on microcarriers even at a relatively high agitation rate if sufficient time was allowed for cell attachment and adhesion. However, if a high agitation rate was applied shortly after initial cell attachment, the growth rate was retarded. This sensitivity to mechanical agitation appears to be dependent on the inoculation cell density.
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  • 28
    ISSN: 1573-0778
    Keywords: apoptosis ; bcl-2 ; cell culture ; chloramphenicol acetyltransferase ; recombinant protein ; Sindbis virus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Viruses carrying foreign genes are often used for the production of recombinant proteins in mammalian cells and other eukaryotic expression systems. Though high levels of gene expression are possible using viral vectors, the host cell generally responds to the infection by inducing apoptotic cell death within several days, abruptly ending protein production. It has recently been demonstrated, however, that apoptosis can be suppressed in virally infected cells using anti-apoptotic genes, such as bcl-2. In this study, stably transfected rat carcinomal cell lines, AT3-bcl2 and AT3-neo, were infected with a Sindbis virus carrying the gene for chloramphenicol acetyltransferase (CAT) in an effort to determine the effect of bcl-2 on cell viability and recombinant protein production. Infected AT3-bcl2 cells consistently maintained viabilities close to 100% and a growth rate equivalent to that of uninfected cells (0.040 h-1). In contrast, the Sindbis viral vector induced apoptosis in the AT3-neo cells, which were all dead by three days post-infection. Though infected AT3-neo cells generated higher levels of heterologous protein, over 1000 mUnits per well, CAT activity fell to zero by two days post-infection. In contrast, chloramphenicol acetyltransferase was present in AT3-bcl2 cells for almost a week, reaching a maximum level of 580 mUnits per well. In addition, recombinant protein production in AT3-bcl2 cells was extended and amplified by the regular addition of virus to the culture medium, a process which resulted in expression for the duration of the cell culture process.
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  • 29
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    Cytotechnology 22 (1996), S. 263-267 
    ISSN: 1573-0778
    Keywords: biodegradable ; bone regeneration ; cell culture ; human cell osteoblasts ; polymers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The care of patients with a skeletal deficiency currently involves the use of bone graft or a non-biologic material such as a metal or polymer. There are alternate possibilities in development which involve the growth of bone cells (osteoblasts) on degradable polymer scaffolds. These tissue engineering strategies require production of the polymeric scaffold, cellular harvest followed by either ex vivo or in vivo growth of the cells on the scaffold, and exploration of the interaction between the cell and scaffold. Research into these strategies utilizes cells from a variety of species, but clinical applications will likely require human osteoblasts. This study explores the process whereby human osteoblasts are harvested under sterile conditions during joint replacement surgery from normally discarded cancellous bone, transported from the operating room to the lab, and grown in culture. This process is feasible, and the cells express their phenotype via the production of alkaline phosphatase and collagen in culture.
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  • 30
    ISSN: 1573-0778
    Keywords: CHO ; IGF-I ; serum-free ; autocrine growth ; cell culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Insulin-like growth factor I has similar mitogenic effects to insulin, a growth factor required by most cells in culture, and it can replace insulin in serum-free formulations for some cells. Chinese Hamster Ovary cells grow well in serum-free medium with insulin and transferrin as the only exogenous growth factors. An alternative approach to addition of exogenous growth factors to serum-free medium is transfection of host cells with growth factor-encoding genes, permitting autocrine growth. Taking this approach, we constructed an IGF-I heterologous gene driven by the cytomegalovirus promoter, introduced it into Chinese Hamster Ovary cells and examined the growth characteristics of Insulin-like growth factor I-expressing clonal cells in the absence of the exogenous factor. The transfected cells secreted up to 500 ng/106 cells/day of mature Insulin-like growth factor I into the conditioned medium and as a result they grew autonomously in serum-free medium containing transferrin as the only added growth factor. This growth-stimulating effect, observed under both small and large scale culture conditions, was maximal since no further improvement was observed in the presence of exogenous insulin.
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  • 31
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    Cytotechnology 3 (1990), S. 157-169 
    ISSN: 1573-0778
    Keywords: acid proteinase ; cell culture ; hybridoma ; immunoglobulin cleavage ; lysosomal proteinases ; recycling reactor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract An acid proteinase has been detected in culture supernate of the 9.2.27 murine hybridoma. This enzyme extensively degrades albumin and transferrin during short incubations at pH 3 and below. Limited proteolysis of the 9.2.27 IgG2a appears to occur in the culture supernate. Proteolysis is enhanced at low pH in the presence of urea or 1 M acetic acid. The proteinase activity accumulates in continuous perfusion, total cell recycle cultures, beginning during exponential growth of the hybridoma. It is destroyed by boiling and blocked by pepstatin, but not by inhibitors of cysteine or serine proteinases or by EDTA. The low pH optimum may distinguish this enzyme from the known rat and mouse aspartic acid proteinases including cathepsin D and cathepsin E.
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  • 32
    ISSN: 1573-0778
    Keywords: cell culture ; polymerase chain reaction ; retrovirus ; reverse transcriptase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Reverse transcriptase (RT) is a good diagnostic tool for the detection of retroviruses. We have developed a simple and rapid assay for RT activity in culture supernatants. A 370-base RNA sequence from the tetracycline-resistance gene in pBR322 plasmid DNA was used as a template for RT-mediated cDNA synthesis. To detect the resultant cDNA, we used the nested polymerase chain reaction. A sensitivity test using purified recombinant RT of human immunodeficiency virus type 1 demonstrated that the detection limit of this method was 10-7–10-8 units of RT activity in 20 μl of a test sample (2 × 10-9–2 × 10-10 units ml-1). This method detected RT activity in unconcentrated supernatants of cell cultures infected with human T-cell leukemia virus, Moloney murine leukemia virus, Moloney murine sarcoma virus, or Rous sarcoma virus. This nonisotopic method provides results within 10 h and is useful for quality control to detect retroviruses in cell cultures.
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  • 33
    ISSN: 1573-0778
    Keywords: cell culture ; cell culture apparatus ; dialysis membrane ; perfusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract We recently developed a new dialysis culture system (termed LIFROC-device) for the cultivation of lymphokine-activated killer cells (Murataet al., 1990, 1991). In the present study, we applied the LIFROC-device (400 ml culture vessel) to the cultivation of mammalian cells for the production of biologically active substances. We cultured mouse-mouse hybridoma TP-709, secreting anti-tissue plasminogen activator (tPA) monoclonal antibody (mAb), recombinant CHO GT19, secreting hGH, and human melanoma Bowes cells, secreting tPA. With the LIFROC-device, TP-709 grew to a maximal cell density of 3.8×106 cells/ml and and produced 480 μg/ml (192 mg in total) of mAb. GT19 reached a cell density of 2.2×106 cells/ml and produced 302 μg/ml (120 mg in total) of hGH. Bowes cells expanded to 4.4×106 cells/ml and secreted 8.5 μg/ml (3.3 mg in total) of tPA. The protein concentration in the culture broths of the LIFROC-device became 7–200 times higher than that of batch culture. Thus, the LIFROC-device can be applied to protein production as well as cell growth with high efficiency.
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  • 34
    ISSN: 1573-0778
    Keywords: cell culture ; colorectal cancer ; microbial contamination ; stroma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Development of primary colorectal cancer cell lines ishampered by contamination from regional microbes, overgrowthof stromal cells, and purported genetic drift from selectionpressures in vitro. We initiated 32 primaryadenocarcinomas, 3 recurrences and 6 distant metastases incell culture. Twelve cell lines from eleven tumors weregenerated (26.8%) overall. Nine of 32 primary tumorsyielded 10 cell lines, 5 were lost to contamination, 13 wereoverwhelmed by stromal cells, and 5 demonstrated no growth.Addition of isobutyl methyl xanthine (IBMX) to culturelimited fibroblastoid growth. There was no associationbetween tumor location (p = 0.535, mid-P), degree ofdifferentiation (p = 0.850, mid-P) or clinicopathologic stage(p = 0.400, mid-P), and the ability of cells to becomeestablished in culture. The majority of cell lines hadsimilar nuclear DNA content and expression of cell-surfaceantigens compared with their parent tumors. Microbialcontamination and stromal cell overgrowth present thegreatest obstacle to capturing a representative bank ofcolon tumors in vitro.
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  • 35
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    Cytotechnology 5 (1991), S. 15-30 
    ISSN: 1573-0778
    Keywords: high density ; cell culture ; serum-free medium ; hybridoma ; CHO cells ; virus production ; insect cells ; adoptive immunotherapy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Conclusion At the 1989 annual meeting of the U.S. Tissue Culture Associations, Ricahrd am, a leading investigator in the serum-free nutrient requirements of cultured cells, commented on the process of medium development. He noted that a survey of major media manufacturers revealed that, among the top selling mammalian cell culture media formulations, most were nearly thirty years old. This commentary is noteworthy considering the tremendous changes in cell culture understanding and derived applications which have emerged over these three decades. Fastidious cell types relatively unknown to investigators of the 1950s and 1960s are now being cultivated in defined, serum-free environments. Culture environments range from limiting dilution clonal recoveries to maintenance cultures approaching tissue densities. While research applications continue to predominate, applications of cell culture have expanded to the engineered production of biopharmaceuticals, to replacement of animal models for toxicology testing, and to the preservation, activation and expansion of human cells, tissues and organs. It is likely that future nutrient medium development will be predicated upon the design of a minimal number of defined formulations of relatively generic utility to a broad class of cell types. Analytical techniques derived from those described herein will be exploited in the user laboratory and in collaboration with the supplier to optimize the nutrient composition for the desired biological response.
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  • 36
    ISSN: 1573-0778
    Keywords: bronchus ; cell culture ; cytology ; morphometry ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Malignant A-549 lung carcinoma and adenovirus-12 SV40 hybrid virus transformed non-tumorigenic human bronchial epithelial cells (BEAS-2B) were objectively discriminated from normal bronchial epithelial (BE) cells on the basis of Papanicolaou stained nuclear features (e.g. shape, chromatin texture, hyperchromasia) and nucleolar morphology (e.g. number per cell, irregular contours). Morphometric analysis indicated that significant differences in cellular morphology existed between BE, BEAS-2B, and A-549 cells. Similar analyses of transformed, tumorigenic cell lines demonstrated that nuclear features (i.e., chromatin texture, clearing of parachromatin, hyperchromasia, variation in thickness of the nuclear envelope, sharp indentations in the nuclear envelope), and nucleolar features (i.e., degree of roundness, presence of angular projections, number per cell) discriminated chemically and virally transformed cells from spontaneously transformed cells. Nuclear and nucleolar features were correlated with the growth rate of tumorigenic cell lines. These analytical approaches will be helpful in studies of the effects of various factors (e.g. vitamin A, phorbol ester, oncogene transfection) on cellular proliferation and/or differentiation.
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  • 37
    ISSN: 1573-0778
    Keywords: proinsulin processing ; CHO ; mutant human proinsulin ; cell culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Heterologous genes encoding proproteins, including proinsulin, generally produce mature protein when expressed in endocrine cells while unprocessed or partially processed protein is produced in non-endocrine cells. Proproteins, which are normally processed in the regulated pathway restricted to endocrine cells, do not always contain the recognition sequence for cleavage by furin, the endoprotease specific to the constitutive pathway, the principal protein processing pathway in non-endocrine cells. Human proinsulin consists of B-Chain — C-peptide — A-Chain and cleavage at the B/C and C/A junctions is required for processing. The B/C, but not the C/A junction, is recognised and cleaved in the constitutive pathway. We expressed a human proinsulin and a mutated proinsulin gene with an engineered furin recognition sequence at the C/A junction and compared the processing efficiency of the mutant and native proinsulin in Chinese Hamster Ovary cells. The processing efficiency of the mutant proinsulin was 56% relative to 0.7% for native proinsulin. However, despite similar levels of mRNA being expressed in both cell lines, the absolute levels of immunoreactive insulin, normalized against mRNA levels, were 18-fold lower in the mutant proinsulin-expressing cells. As a result, there was only a marginal increase in absolute levels of insulin produced by these cells. This unexpected finding may result from preferential degradation of insulin in non-endocrine cells which lack the protection offered by the secretory granules found in endocrine cells.
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  • 38
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    Cytotechnology 30 (1999), S. 149-158 
    ISSN: 1573-0778
    Keywords: bioreactor ; cell culture ; disposable ; wave agitation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract This work describes a novel bioreactor system for the cultivation of animal, insect, and plant cells using wave agitation induced by a rocking motion. This agitation system provides good nutrient distribution, off-bottom suspension, and excellent oxygen transfer without damaging fluid shear or gas bubbles. Unlike other cell culture systems, such as spinners, hollow-fiber bioreactors, and roller bottles, scale-up is simple, and has been demonstrated up to 100 L of culture volume. The bioreactor is disposable, and therefore requires no cleaning or sterilization. Additions and sampling are possible without the need for a laminar flow cabinet. The unit can be placed in an incubator requiring minimal instrumentation. These features dramatically lower the purchase cost, and operating expenses of this laboratory/pilot scale cell cultivation system. Results are presented for various model systems: 1) recombinant NS0 cells in suspension; 2) adenovirus production using human 293 cells in suspension; 3) Sf9 insect cell/baculovirus system; and 4) human 293 cells on microcarrier. These examples show the general suitability of the system for cells in suspension, anchorage-dependent culture, and virus production in research and GMP applications.
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  • 39
    ISSN: 1573-0778
    Keywords: cell culture ; carcinoembryonic antigen ; aspirin ; enhanced production ; Langmuir-Blodgett film
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Human colorectal adenocarcinoma tumor (CW2) cells were cultivated in RPMI 1640 media containing 0–7.5 mM aspirin and 10% fetal bovine serum for the production of carcinoembryonic antigen (CEA). By adding aspirin to the media, the production of CEA per cell increased by up to one hundred fold compared to cultivation in normal media containing no aspirin, even though the total cell concentration decreased with the increase in aspirin in the media. The production of CEA was also investigated for CW2 cells cultured on silk fibroin, poly(γ-benzyl-L-glutamate) and poly(γ-benzyl-L-glutamate)/poly(ethylene oxide) diblock copolymer films prepared by the Langmuir-Blodgett and casting methods. The highest production of CEA per cell was observed for the CW2 cells on poly(γ-benzyl-L-glutamate) and its diblock copolymer films prepared by the Langmuir-Blodgett method in the medium containing 5 mM aspirin after 168 hr of inoculation. This originates from the fact that the cell density on the films in the medium containing 5 mM aspirin was the lowest under these conditions. It is suggested that CW2 cells produce CEA more effectively when the cell growth is suppressed by addition of toxic chemicals such as aspirin or by culture on unfavorable films for cell growth.
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  • 40
    ISSN: 1573-0778
    Keywords: cell culture ; hollow fiber bioreactor ; hybridoma ; micro bioreactor ; optimization ; T-flask
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract In this article, cell growth in a novel micro hollow fiberbioreactor was compared to that in a T-flask and theAcuSyst-Maximizer®, a large scale industrial hollowfiber bioreactor system. In T-flasks, there was relativelylittle difference in the growth rates of one murine hybridomacultured in three different media and for three other murinehybridomas cultured in one medium. However, substantialdifferences were seen in the growth rates of cells in themicro bioreactor under these same conditions. These differencecorrelated well with the corresponding rates of initial cellexpansion in the Maximizer. Quantitative prediction of thesteady-state antibody production rate in the Maximizer was moreproblematic. However, conditions which lead to faster initialcell growth and higher viable cell densities in the microbioreactor correlated with better performance of a cell line inthe Maximizer. These results demonstrate that the microbioreactor is more useful than a T-flask for determining optimalconditions for cell growth in a large scale hollow fiberbioreactor system.
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  • 41
    ISSN: 1573-0778
    Keywords: anchorage-dependent cell ; cell culture ; packed-bedreactor ; retroviral vector ; viral production
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract To indentify a plausible large-scale production system forretroviral vector, three culture systems, i.e., batch culturewith medium exchange, microcarrier culture, and packed-bedreactor culture were compared. In batch cultures with mediumexchange, high cell concentrations were maintained for about amonth, and the harvested retroviral titer remained constant. Inmicrocarrier cultures, although cell growth was rapid, theretroviral titer was unexpectedly low, suggesting that the lowtiter was due either to serious damage to the retroviral vectoror to a reduction in the production rate of retroviral vector,caused by mechanical shear forces. Although the retroviral titer(maximum titer, 1.56 × 106) in the packed-bedreactor was a little bit lower than that obtained in the batchculture with medium exchange (maximum titer, 1.91 ×106), continuous production made it possible to increasethe cumulative titer up to 16-fold of that from the batchculture with medium exchange. Moreover, as the packed-bedreactor system requires less labor and shows excellentvolumetric productivity in comparison to batch cultures withmedium exchanges, it will be an appropriate production systemfor retroviral vector in large quantities.
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  • 42
    ISSN: 1573-0778
    Keywords: acridine derivative ; cell culture ; fluorescence microscopy ; mycoplasma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A new fluorescent acridine orange derivative, 3-amino-6-methoxy-9-(2-hydroxyethylamino) acridine (AMHA), has been applied to Hela cells in order to set up appropriate conditions for the detection of mycoplasma contaminations. Since AMHA staining reveals intensely fluorescent nuclei and slight fluorescent cytoplasm, we can visualize and localize mycoplasma contamination on each cell. In combination with a shortened Chen's staining method (1977), AMHA should allow a better detection of mycoplasma in animal cell cultures than the well established Hoechst dye.
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  • 43
    ISSN: 1573-0778
    Keywords: cell culture ; kinetics ; Ig promoter/enhancer ; plasmacytoma ; recombinant protein production
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A model mammalian cell system for the production of recombinant proteins was investigated. Murine myeloma cells which had lost the ability to produce both heavy and light chain immunoglobulin molecules were transfected with a vector containing the immunoglobulin heavy chain promoter and enhancer elements linked to the human growth hormone gene. The growth kinetics of G32, a clonal isolate, were found to be similar to both the parent myeloma and hybridomas. However, production of hGH by G32 was growth associated, rather than as a secondary metabolite as is the case for hybridomas. In addition, G32 produced hGH at molar levels greater than most hybridomas.
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  • 44
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    Cytotechnology 6 (1991), S. 49-54 
    ISSN: 1573-0778
    Keywords: Bivalvia ; cell culture ; embryo ; mitosis ; scallop
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Primary cell cultures obtained from embryos of Mizuchopecten yessoensis (Bivalvia) survived for four months. Although the number of cells progressively decreased during the cultivation, mitotic cells were observed both at the first stages and at the end. A possibility of growing marine invertebrates cells in long term primary culture is discussed.
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  • 45
    ISSN: 1573-0778
    Keywords: cell culture ; endosteal human osteoblasts ; maxilla ; mandible ; titanium ; biocompatibility ; alkaline phosphatase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Fragments of cancellous and cortical bone from human maxilla and mandible were cultured by the explant technique. Cells isolated by trypsinization of primary cultures were characterized as osteoblasts on the basis of intracellular alkaline phosphatase activity, the constituents of the extracellular matrix, and response to human parathormone (PTH). In culture, the osteoblasts often gave rise to superposed clumps of large cells whose cytoplasm contained endoplasmic reticulum, numerous mitochondria, vacuoles, and a dense network of intermediate filaments, often at the level of the plasma membrane. In the presence of vitamin C and 1,25-dihydroxyvitamin D3, the osteoblasts produced an extracellular matrix composed of collagen type I and various non-collagenous proteins, including osteocalcin. Biochemical test results were comparable to those reported for osteoblasts of other origins (rat calvaria, human iliac crest), and namely elevated intracellular alkaline phosphatase activity and cAMP accumulation in response to stimulation by human PTH (1–34). Osteoblasts isolated in this manner were cultured in the presence of pure titanium disks to determine the effects of exposure to this metal. Electron microscopy revealed few significant differences in cell growth and specific enzyme activity compared to control osteoblasts grown on plastic dishes, reflecting the excellent biologic and biochemical relationship between the osteoblasts and pure titanium. This experimental system thus appears suitable for biocompatibility studies, and in particular, evaluation of dental implants.
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  • 46
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    World journal of microbiology and biotechnology 15 (1999), S. 635-637 
    ISSN: 1573-0972
    Keywords: Anthocyanin ; cell culture ; conditioned medium ; strawberry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A conditioned medium (CM) prepared from strawberry suspension cultures greatly stimulated anthocyanin accumulation. CM separated by dialysis membrane showed a significant increase (p 0.05) in anthocyanin synthesis at a fraction smaller than 10,000 Da. The stimulation by CM was eliminated when the CM was treated with alkali.
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  • 47
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    AIChE Journal 35 (1989), S. 1177-1185 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A theoretical and experimental study has been carried out on flow, dissolution, and precipitation in porous media. Flow experiments were performed on linear carbonate cores using acidic ferric chloride solutions. Dissolution of the carbonate by the acid causes an increase in the solution pH, thereby precipitating ferric hydroxide. This precipitate plugs up the pore throats in the medium and increases the resistance to fluid flow. Fluctuations in the permeability ratio were observed during core flood experiments, confirming the competition between channel formation due to dissolution and pore plugging due to precipitation. The evolution of the pore structure was characterized by Wood's metal castings.A network model has also been developed to describe flow and reaction in porous media. The model was used to simulate the ferric chloride system, and pressure oscillations predicted by the model show identical trends to those observed experimentally. Additionally, the evolution of pores in the network were graphically represented.
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  • 48
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    AIChE Journal 35 (1989), S. 988-994 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The extent of high temperature (900-1,300°C), short time (〈1 s) SO2 capture was found to be limited by temperature-dependent losses in the porosity of calcium based sorbents. At 970°C these porosity losses were caused by CO2-activated sintering. Sulfation of the sorbents either prevented or reduced the extent of porosity losses. Differences in SO2 capture between hydroxides from different commercial sources, and significantly lower levels of capture by calcium carbonates compared to hydroxides were attributed to differences in particle size and the degree of porosity loss.
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  • 49
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    AIChE Journal 35 (1989), S. 1017-1028 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Systems having transfer functions of the form \documentclass{article}\pagestyle{empty}\begin{document}$$ G_P (s) = \frac{{P_1 (s) - P_2 (s)e^{ - t_d s} }}{{Q(s)}}, $$\end{document} where P1(s), P2(s) and Q(s) are polynomials, are called quasirational distributed systems (QRDS). They are encountered in processes modeled by hyperbolic partial differential equations. QRDS can have an infinity of right half-plane zeros which causes large phase lags and can result in poor performance of the closed-loop system with PID controllers. Theory on the asymptotic location of zeros of quasipolynomials is used to predict the nonminimum phase characteristics of QRDS and formulas are presented for factoring QRDS models into minimum and non-minimum phase elements.A generalized Smith predictor controller design procedure for QRDS, based on this factorization, is derived. It uses pole placement to obtain a controller parameterization that introduces free poles which are selected to satisfy robustness specifications. The use of pole placement allows for the design of robust control systems in a transparent manner. Controller selection is generally better, simpler and more direct with this procedure than searching for optimal PID controller settings.
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    AIChE Journal 35 (1989), S. 1049-1052 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
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    AIChE Journal 35 (1989), S. 1053-1053 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
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    AIChE Journal 35 (1989), S. 1097-1106 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The sorption of toluene, dilute in CO2, and likewise the distribution coefficient of toluene between silicone rubber and CO2 have been explored from gaseous to supercritical fluid conditions. Sorption and desorption isotherms were determined by frontal analysis using a new inverse supercritical fluid chromatography technique at 35 and 70°C up to 250 bar. Complementary swelling data are presented for pure CO2 in silicone rubber up to 315 bar. A new result is that the sorption of toluene goes through a maximum and is highly adjustable over a continuum in the highly compressible region of carbon dioxide. This behavior is explained physically and predicted quantitatively with the Flory equation and the Peng-Robinson equation of state using only information from binary systems. These results are useful for a wide variety of applications including impregnation of polymers with pharmaceuticals, fragrances and other additives, and polymer purification.
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    AIChE Journal 35 (1989), S. 1137-1147 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Detailed mathematical models, pore or grain based, describing diffusion, reaction, and pore structure evolution in pellets of porous solids are used to simulate, along with a macroscopic reactor design model, the transient phenomena that take place during desulfurization of coal gas in fixed-bed reactors of pellets of metal oxide sorbents. Our computer simulation results show that the form of the pore- or grain-size distribution strongly influences the predictions of the overall reactor design model.
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    AIChE Journal 35 (1989), S. 1186-1194 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
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    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Vapors of alkali metal compounds can be removed from coal combustion and gasification flue gases using high-temperature aluminosilicate sorbents. The fundamentals of alkali adsorption on kaolinite, bauxite, and emathlite are compared and analyzed both experimentally and through theoretical modeling. The results show that the process is not a simple physical condensation, but a complex combination of diffusion and reaction. The kinetics of adsorption on these sorbents have similarities: the process is diffusion-influenced, the rate decreases with time, and there is a final saturation limit. There are, however, differences in reaction mechanisms leading to potentially different applications for each sorbent. In adsorbing alkali chloride vapors, kaolinite and emathlite release all the chlorine back to the gas phase while bauxite retains some of the chlorine. Moreover, the products of reaction with emathlite have a melting point significantly lower than those for kaolinite and bauxite. Therefore, emathlite is more suitable for lower-temperature sorption systems downstream of the combustors/gasifiers, while kaolinite and bauxite are suitable as in-situ additives.
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    AIChE Journal 35 (1989), S. 1203-1206 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
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    AIChE Journal 35 (1989), S. 1210-1210 
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    Keywords: Chemistry ; Chemical Engineering
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    AIChE Journal 35 (1989), S. 1351-1364 
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    Keywords: Chemistry ; Chemical Engineering
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    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In the present investigation, Taylor's analysis of the axial dispersion of a solute in a Newtonian fluid undergoing laminar flow through a circular tube was applied to dispersions of colloidal particles, in which effects of size exclusion, inertial and colloidal forces, and wall retardation must be considered. The results indicate that the product of the particle Reynolds and Peclet numbers determines the importance of the inertial forces on both the effective axial diffusion coefficient and the height of a theoretical plate.The height of a theoretical plate as a function of the eluant ionic strength and average velocity, particle diameter, and tube diameter was determined experimentally. Close agreement with the numerical calculations from the diffusion equation was obtained. The height of a theoretical plate was found to attain a maximum value when the product of Reynolds and Peclet numbers was approximately 10.5.
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    AIChE Journal 35 (1989), S. 1385-1390 
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    Keywords: Chemistry ; Chemical Engineering
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    AIChE Journal 35 (1989), S. 1402-1403 
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    Keywords: Chemistry ; Chemical Engineering
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    AIChE Journal 35 (1989), S. 1403-1404 
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    Keywords: Chemistry ; Chemical Engineering
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    AIChE Journal 35 (1989), S. 1337-1345 
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    Keywords: Chemistry ; Chemical Engineering
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    Notes: The recent model of partial internal wetting of catalyst particles (Bhatia, 1988) is extended to allow for pore-size distribution, multicomponent diffusion, and nonlinear kinetics for spherical particles exposed to condensible vapor undergoing an exothermic reaction. Profiles of liqid filling of the pores are computed, and the influence on mass transfer and effectiveness factor is determined. Under certain circumstances, the effectiveness factor has a maximum with respect to the mole fraction of condensible component suggesting an optimum vapor-phase composition. As the bulk vapor approaches saturation the effectiveness factor and particle temperature can drop sharply because of precipitious increase in liquid filling. However, a significant fraction of the pore space is still dry in contradiction to prior models assuming complete internal wetting of catalyst particles. The new model is more realistic than earlier attempts and lays the framework for proper representation of the physical phenomena involved.
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    AIChE Journal 35 (1989), S. 1346-1350 
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    Keywords: Chemistry ; Chemical Engineering
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    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Andritsos and Hanratty (1978b) have shown that the increase in interfacial drag caused by waves in stratified gas-liquid flows is related to the wave steepness. Recent analyses of finite amplitude Kelvin-Helmholtz waves are used to develop a correlation for the ratio of the wave height to wavelength.
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    AIChE Journal 35 (1989), S. 1328-1336 
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    Notes: The solid circulation in the annular region of a 0.146 and a 0.292 m ID semicylindrical and cylindrical air spouted beds was investigated for different spouting conditions. Stroboscopic photography, stopwatch and fiber optic methods were used to measure the vertical particle velocities in the annular region. The optical fiber probe enabled the measurement of particle velocities inside the dense annular phase and was validated at the walls. Velocity profiles in half beds were very different from those found in full beds. The work was then limited to full beds. Experimental results show that the solids flow is characterized by a point sink at the nozzle entrance with the solids moving almost in plug flow higher in the bed. There is negligible entrainment of solids along the spout wall and slow, thin wall layers are observed near the walls and spout. The velocity profiles are independent of the total height of the bed. A kinematic model successfully describes the observed velocity field.
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    AIChE Journal 35 (1989), S. 1535-1542 
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    Keywords: Chemistry ; Chemical Engineering
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    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Many industrially important processes feature both nonlinear system dynamics and a process deadtime. Powerful deadtime compensation methods, such as the Smith predictor structure in state space for linear systems is presented first and then directly extended to nonlinear systems. When combined with input/output linearizing state feedback, this Smith-like predictor makes a nonlinear system with deadtime behave like a linear system with deadtime. The control structure is completed by adding an external linear controller, which provides integral action and compensates for the deadtime in the input/output linear system, and an open-loop state observer. Conditions for robust stability with respect to errors in the deadtime and more general linear unstructured multiplicative uncertainties are given. Computer simulations for an example system demonstrate the high controller performance that can be obtained using the proposed method.
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    AIChE Journal 35 (1989), S. 1543-1546 
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    Keywords: Chemistry ; Chemical Engineering
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    AIChE Journal 35 (1989), S. 1569-1571 
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    Keywords: Chemistry ; Chemical Engineering
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    AIChE Journal 35 (1989) 
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    AIChE Journal 35 (1989), S. 1617-1624 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
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    Notes: Ultrapyrolysis or ultrarapid pyrolysis is a hydrocarbon thermal cracking process which offers the promise of greater product selectivity, higher yield and feedstock flexibility. In this paper, ultrapyrolysis of propane in a spouted bed with a draft tube was used as a test system to demonstrate these advantages. Experiments, carried out on a 20-cm-diameter pilot-scale reactor, illustrate the spouted bed to be capable of achieving the extremely high reactant heating rates of up to 105 K/s and the short gas residence times of less than 500 ms necessary for this process. Moreover, reactant conversion and product yields can be enhanced by controlling the operating temperature of the bed. In addition, application of the propane pryolysis reaction scheme of Sundaram and Froment into a recently developed computer model indicates the ability to correctly simulate the spouted bed as an ultrapyrolysis reactor.
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    AIChE Journal 35 (1989), S. 1651-1658 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
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    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This paper focuses on the problem of scale-up of two-phase (e.g., solid-fluid) reactors. It outlines a class of such reactors which, for first-order reactions, scale in the familiar single-phase manner via an analog of the usual residence time distribution formula. The differences lie in the fact that the appropriate tracer experiment entering the formalism for this reactor class refers to a nonadsorbing tracer, and the analog of the plug flow solution is the solution for a fixed bed with fluid-phase plug flow. Surprisingly, unlike single-phase systems, there exist two-phase reactors, outside of the class defined, that do not scale in this manner, even when, say, the true catalyst surface chemistry is purely first-order. The paper discusses a few examples and implications for the design of two-phase reactors, including fluidized beds.
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    AIChE Journal 35 (1989), S. 1685-1691 
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    Keywords: Chemistry ; Chemical Engineering
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    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Heat transfer coefficients were measured for ciculating beds of sand particles of mean size 222 to 299 μm at temperatures of 340-880° C. Transfer coefficients were obtained for both a 1.22-m-long, 12.7-mm-OD vertical tube and a 1.59-m-long, 148-mm-wide membrane wall near the top of a 152-mm-square by 7.32-m-tall combustion column. For both surfaces and all temperatures, average heat transfer coefficients increased almost linearly with local suspension density which ranged from 0 to 70 kg/m3. Radiation played a significant role, especially at high temperatures and low suspension densities. Heat transfer coefficients also varied significantly with the lateral position of the tube. The vertical length of heat transfer surface is shown to be an important parameter allowing seemingly discrepant published results to be reconciled.
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    AIChE Journal 35 (1989), S. 1719-1727 
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    Keywords: Chemistry ; Chemical Engineering
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    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An approximate equation for the evolution of finite-amplitude, long disturbances to Newtonian liquid films is found to be accurate over a wide range of Reynolds numbers. A long-wave expansion leads to a film profile equation asymptotically correct when wave number and Reynolds number are small. Solutions of the film profile equation are compared to exact and other approximate solutions of the Navier-Stokes system. An alternative form of the film profile equation results in remarkably accurate solutions, when Reynolds numbers are moderate, in the cases of standing or monotonically decaying waves in horizonatal films, rising film flow, final acceleration of a moving film, and film flow emerging from a slot coater.
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    AIChE Journal 35 (1989), S. 1742-1744 
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    Keywords: Chemistry ; Chemical Engineering
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    AIChE Journal 35 (1989), S. 1753-1756 
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    Keywords: Chemistry ; Chemical Engineering
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    AIChE Journal 35 (1989), S. 1779-1790 
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    Keywords: Chemistry ; Chemical Engineering
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    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The kinetics of protein aggregation in salt-induced precipitation processes were studied as a function of salt type, salt concentration, temperature and protein concentration. α-chymotrypsin (αCT) was used as a model protein. Stopped-flow turbidimetry was used to monitor the progress of precipitations. Analysis of the linear portions of the turbidity trajectories indicates that temperature and salt concentration effects are related to protein solubility; the protein concentration dependence is well described by the Smoluchowski collision equation. The aggregation kinetics of partially-inhibited αCT exhibit poisoning behavior, underscoring the importance of dimerization and monomer addition in the precipitation of αCT. Solute particle radius distributions determined via dynamic laser light scattering for low salt and supernatant αCT solutions indicated that significant aggregation does not occur in the absence of supersaturation. A detailed population balance model was proposed that accounted for specific and nonspecific interactions and monomer addition. The model is expected to find general application to protein aggregation phenomena, in particular for proteins that have specific quaternary interactions.
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    AIChE Journal 35 (1989), S. 1829-1834 
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    Notes: A low residence time flow method for the measurement of the critical temperatures and pressures of thermally-unstable fluids was developed in this work. The low residence time at elevated temperatures minimizes decomposition and other reactions, making it possible to measure the critical properties of many unstable fluids. An extrapolation procedure was developed to obtain the critical properties of rapidly reacting substances, based on the linear behavior of the apparent critical property with residence time. The measured critical properties of 14 n-alkanes (pentane through octadecane) using this method are reported in this paper. The results extend the available data on the critical properties of the higher alkanes and reveal a possible error in the literature value for the critical pressure of tetradecane.
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    AIChE Journal 35 (1989), S. 1835-1844 
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    Keywords: Chemistry ; Chemical Engineering
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    Notes: A feedforward/feedback version of a single input/single output self-tuning controller has been developed and tested both via simulation studies and experiments on a section of an absorption/desorption pilot plant. The algorithm is based on the recursive least squares estimation of parameters for the linear models relating the output to the controlled input and to the disturbances; adaptation is achieved using a variable forgetting factor. The control input at each time interval is calculated using one of several single-step extended-horizon control strategies.The results show that the performance of the algorithm is insensitive to the choice of initial parameters, all of which have a readily identifiable intuitive basis. The algorithm is especially robust against deterministic disturbances (measurable and unmeasurable) and unknown and varying time delays. Computational load beyond that of a feedback-only version is minimal.
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    AIChE Journal 35 (1989), S. 1869-1875 
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    Keywords: Chemistry ; Chemical Engineering
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    Notes: Equilibrium isotherms and kinetic data for sorption of ethylamine on two different H-form ion exchangers (a homogeneous gel type and a biporous MR type) have been determined experimentally by the gravimetric method. The gaseous amine is adsorbed on the dry resins according to an acid-base neutralization reaction, and the saturation capacity coincides with the exchange capacity of the resin. For the gel-type resin, the equilibrium is less favorable and the isotherm is almost linear over the experimental range.The kinetic data also reflect the structural difference between the two adsorbents. In the MR-type resin, the sorption rate is controlled by macropore diffusion with rapid equilibration of the adsorbed phase within the microparticles. Since the equilibrium isotherm is highly favorable, approaching the irreversible limit, the uptake curves are well represented by the ‘shrinking core’ model. Diffusion in the gel-type resin is much slower and the pore diffusivities are smaller by several orders of magnitude, presumably reflecting the smaller effective pore diameter.These results suggest that the MR-type resin is a potentially useful adsorbent for removal of traces of light amines from industrial gases.
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    AIChE Journal 35 (1989), S. 1903-1906 
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    AIChE Journal 35 (1989), S. 1915-1916 
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    AIChE Journal 35 (1989), S. 1927-1932 
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    Notes: The model of uniform nonlinear kinetics recently developed for the continuous description of a multicomponent mixture reacting in a batch or plug flow reactor (Astarita, 1989) is applied to the analysis of a continuous stirred tank reactor (CSTR). It is shown that most of the equations can be solved in general, independently of the specific kinetic mechanism. Some specific mechanisms are discussed. The difference between the behavior in a plug flow reactor (PFR) and a CSTR turns out to be quite dissimilar from what one would expect by considering the single-component case.
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    AIChE Journal 35 (1989), S. 1899-1902 
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    AIChE Journal 35 (1989), S. 1933-1941 
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    Notes: Many industrial proceses require the circulation of particles between reacting fluidized compartments at a stable, but flexible rate, so that the processes can be operated at different solids rates. In order to understand the phenomena involved in the circulation and its control, circulation of sand particles has been studied through a new experimental approach, using an open-loop two compartment circulating system.Experimental results show that the control of the circulation rate depends on three phenomena: the vertical resisting force in the fluidized compartments, the contraction of flow in the communication zone, and the bypass of gas between the compartments. For industrial purposes, the first appears to be predominant in the control of the solids rate. The second ensures neither the strict control of this rate nor the required flexibility. The link between the gas bypass and the circulation phenomena is explained and recommendations for the design and operation of circulating systems are given.
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  • 85
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    AIChE Journal 35 (1989), S. 2037-2039 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
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  • 86
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    Hoboken, NJ : Wiley-Blackwell
    AIChE Journal 36 (1990) 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
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  • 87
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    AIChE Journal 36 (1990), S. 13-18 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Flow-induced crystallization experiments are made in a capillary apparatus modified with a downstream reservoir under pressure. Capillary length, diameter, and entrance angle are changed, as well as flow rate. The results show that the crystallization temperature is influenced both by the elongational flow at the capillary entrance and by the shear flow along the capillary. The independent effect of the pressure equals that obtained under static conditions. The effect of shear is correlated in terms of shearing work.
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  • 88
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    Hoboken, NJ : Wiley-Blackwell
    AIChE Journal 36 (1990), S. 3-12 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A method to calculate the compression zone height in continuous thickeners is presented. With this method, it is necessary to know the variations of the pulp-supernatant interface and the sediment height vs. time in a batch test. This method is discussed considering the following aspects: relationship between the settling rate of solids and their concentration in the settling zone; and the compression of solids due to the squeeze transmitted by the upper layers - unbuoyed weight of particles minus force of friction due to the Darcian flow. When the variation of the sediment height vs. time becomes linear, it is possible to calculate the maximum solids concentration which can be reached by sedimentation. The change of the solids matrix permeability and its influence on the method proposed are also analyzed.
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  • 89
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    Hoboken, NJ : Wiley-Blackwell
    AIChE Journal 36 (1990), S. 29-38 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A kinetic model is developed for the drying of solids in fluidized beds, assuming a falling rate period following a constant rate period. Experimental data obtained using batch and continuous single and spiral fluidized beds are satisfactorily matched with the assumed drying kinetics and the residence time distribution of solids appropriate for the type of dryer.
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  • 90
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    Hoboken, NJ : Wiley-Blackwell
    AIChE Journal 36 (1990), S. 53-65 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The backfill cycle in the pressure swing adsorption (PSA) separation of air to produce an enriched oxygen product using a zeolite 5A molecular sieve was studied theoretically and experimentally. The effects of the backfill cycle, cycle configuration, backfill rate, pressurisation rate, and product rate were studied. The theory agreed well with the experimental results in predicting the product oxygen concentration over a wide range of backfill pressures, thus giving a basic understanding of the bed dynamics of the backfill cycle. The theory showed that the backfill rate and adsorption capacity of the adsorbent were the most important parameters governing the efficiency of the PSA process. We showed that, given an adequate backfill pressure, a separate pressurization step could be omitted by incorporating it with the product release step with no detrimental effect on the product oxygen concentrations while increasing adsorbent productivity.
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  • 91
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    Hoboken, NJ : Wiley-Blackwell
    AIChE Journal 36 (1990), S. 93-105 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Bubble sizes were measured for molten wax-nitrogen systems using photography and dynamic gas disengagement. The effects of operating conditions, system geometry and wax type were studied in 0.05- and 0.23-m-diameter by 3-m-tall bubble columns. Both techniques were used with FT-300 wax, while only the dynamic gas disengagement technique could be used with reactor waxes due to their dark color. For FT-300 wax, Sauter mean diameters obtained from photographs taken near the column wall were significantly lower than those obtained from photographs taken near the center. The ds values obtained from dynamic gas disengagement and photographic (near the column center) methods, for this noncoalescing medium, were in the range 0.5-1.6 mm in the large-diameter column. For reactor waxes, ds values were significantly higher (1-2 mm for Sasol and 1-5.5 mm for Mobil's reactor wax) and are in agreement with results reported in earlier studies with similar waxes (ds = 2-4 mm), where different experimental techniques (light transmission or hot wire anemometry) were employed.
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  • 92
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    Hoboken, NJ : Wiley-Blackwell
    AIChE Journal 36 (1990), S. 132-136 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 7 Ill.
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  • 93
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
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  • 94
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    Hoboken, NJ : Wiley-Blackwell
    AIChE Journal 36 (1990), S. 175-186 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A model has been developed for steady polymer melt blowing. This model includes the dominant effect that the forwarding air has upon the process. Inertial, gravitational and heat transfer effects are also included. The model equations are solved numerically with both Newtonian and viscoelastic (Phan-Thien and Tanner) constitutive equations. The predicted results compare favorably with actual experimental data.
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  • 95
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    Hoboken, NJ : Wiley-Blackwell
    AIChE Journal 36 (1990), S. 227-232 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Instantaneous chemical reactions, either themselves or together with chemical indicators, have previously been used as probes of turbulent micromixing. Both methods obtain statistics of the micromixing from measurement of time mean quantities. The mean reactant concentration, which is the quantity measured in the instantaneous reaction method, is shown here to be the integral with respect to feed concentration ratio of the mean color density, which is the quantity measured in the chemical indicator method. Differentiation of the mean color density gives the scalar probability density while integration gives the mean reactant concentration. Measurements of the color density of bromothymol blue at the centerline of a turbulent jet of base mixing with acid are used as an example.
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  • 96
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    Hoboken, NJ : Wiley-Blackwell
    AIChE Journal 36 (1990), S. 287-290 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 4 Ill.
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  • 97
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    Hoboken, NJ : Wiley-Blackwell
    AIChE Journal 37 (1991), S. 111-122 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The time-averaged void fraction, pressure drop and flow regime transition behavior of horizontal air-water two-phase flows is studied experimentally and numerically for 2-cm-inner-diameter tubes with various flow dividing junctions at its end. The time-average void and pressure drop behavior along the channel is simulated using a two fluid separated flow model. The results show that two-phase behavior (flow regime, void fraction, and pressure drop) is affected strongly by the presence of a flow division in the system. These effects extend far upstream of the junction for low-momentum flows and far downstream for high-momentum flows. Both numerical and experimental results show that there occurs a large increase in void just downstream of the junction owing to the halving of the fluid volume flow rates and the liquid deceleration.
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  • 98
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    Hoboken, NJ : Wiley-Blackwell
    AIChE Journal 37 (1991), S. 137-141 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 1 Ill.
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  • 99
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    Hoboken, NJ : Wiley-Blackwell
    AIChE Journal 37 (1991) 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
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  • 100
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    Hoboken, NJ : Wiley-Blackwell
    AIChE Journal 37 (1991), S. 169-181 
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Nonequilibrium diffusion behavior in nonionic and ionic surfactant microemulsion systems has been studied experimentally using the open-ended capillary method. Experimental results for these systems have been compared with a drop theory of diffusion for microemulsions under conditions where large concentration and electrostatic gradients exist. The results show good agreement in concentration profiles between theory and experiment for the microemulsion components - water, benzene, and phenol. Furthermore, under certain conditions the theory predicts that over a limited time interval phenol will diffuse from low-concentration regions to regions of higher concentration. This phenomenon has been observed.
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