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  • Cell & Developmental Biology  (9)
  • Immobilization  (4)
  • 1990-1994  (13)
  • 1
    Electronic Resource
    Electronic Resource
    Continuous limonin degradation by immobilized Rhodococcus fascians cells in K-carrageenan (1994)
    Springer
    Applied microbiology and biotechnology 41 (1994), S. 487-493 
    Details
    ISSN: 1432-0614
    Keywords: Key words Limonin ; Debittering ; Immobilization ; Rhodococcus fascians ; Continuous Stirred Tank Reactor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Limonin can be effectively degraded by Rhodococcus fascians cells. These bacteria can be entrapped in κ-carrageenan, and used in a continuous stirred tank reactor to degrade limonin in a continuous process. The effects of temperature, limonin concentration, dilution rate, and aeration on the reactor behaviour have been tested, and the results correlated with changes in limonin conversion, substrate degradation rate, and free and immobilized biomass. Results showed that the immobilized cells were able to debitter limonin-containing media and the immobilized biomass was quite stable throughout the operational conditions tested. A population of free biomass was present in the reactor, the quantity of which was dependent on dilution rate. The immobilized bacteria increased its limonin-degrading capability when the substrate concentration was increased. The aeration was not strictly necessary for limonin degradation. Additionally, the immobilized cells were active and stable for more than 2 months of continuous operation, and were able to recover their limonin-degrading capability when used intermittently. Finally, none of the main components of a juice was noticeably altered during limonin degradation, so the reactor response was good enough to consider its application.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01982541
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  • 2
    Electronic Resource
    Electronic Resource
    Effect of glucose on pHin and [Ca2+]in in NIH-3T3 cells transfected with the yeast P-Type H+-ATPase (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 161 (1994), S. 129-141 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: NIH-3T3 cells transfected with yeast H+-ATPases (RN1A cells) are tumorigenic (Perona and Serrano, 1988, Nature, 334: 438). We have previously shown that RN1a cells maintain a chronically high intracellular pH (pHin) under physiological conditions. We have alsoshown that RN1a cells are serum-independent for growth, maintain a higher intracellular Ca2+(in), and glycolyze more rapidly than their non-transformed counterparts (Gillies et al., Proc. Natl. Acad. Sci., 1990, 87: 7414; Gillies et al., Cell. Physiol. Biochem., 1992, 2: 159). The present study was aimed to understand the interrelationships between glycolysis, pHin, and [Ca2+]in in RN1a cells and their non-transformed counterparts, NIH-3T3 cells. Our data show that the higher rate of glycolysis observed in RN1a cell is due to the presence of low affinity glucose transporters. Consequently, the higher rate of glycolysis is exacerbated at high glucose concentration in RN1a cells. Moreover, the maximal velocity (Vmax) for glucose utilization is up to sixfold higher in RN1a cells than in the NIH-3T3 cells, suggesting that the number of glucose transporters is higher in RN1a than NIH-3T3 cells. Glucose addition to NIH-3T3 cells results in modest decreases in both pHin and [Ca2+]in. In contrast, RN1a cells respond to glucose with a large decrease in pHin, followed by a large decrease in [Ca2+]in. The decrease in [Ca2+]in observed upon glucose addition is likely due to activation of Ca2+-ATPase by glycolysis, since the Ca2+ decrease is abolished by the Ca2+ ATPase inhibitors thapsigargin and cyclopiazonic acid. Glucose addition to ATP-depleted cells results in a decrease in [Ca2+]in, suggesting that ATP furnished by glycolysis is utilized by this pump. © 1994 Wiley-Liss, Inc.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041610116
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  • 3
    Electronic Resource
    Electronic Resource
    NIH 3T3 cells transfected with a yeast H+-ATPase have altered sensitivity to insulin, insulin growth factor-I, and platelet-derived growth factor-AA (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 159 (1994), S. 551-560 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The role of intracellular pH (pHin) in the regulation of cell growth in both normal and transformed cells is a topic of considerable controversy. In an effort to study this relationship NIH 3T3 cells were stably transfected with the gene for the yeast H+-ATPase, constitutively elevating their pHin. The resulting cell line, RN1a, has a transformed phenotype: The cells are serum independent for growth, clone in soft agar, and form tumors in nude mice. In the present study, we further characterize this system in order to understand how transfection with this proton pump leads to serum-independent growth, using defined media to investigate the effects of specific growth factors on the transfected and parental NIH 3T3 cells. While both cell lines show similar growth increases in response to platelet-derived growth factor (PDGF)-BB and epidermal growth factor (EGF), they respond differently to insulin, insulin-like growth factor-I (IGF-I) and PDGF-AA. RN1a cells exhibit increased growth at nanomolar concentrations of insulin but the parental cells had only a relatively minor response to insulin at 10 μM. Both cell lines showed some response to IGF-I in the nanomolar range but the response of RN1a cells was much larger. Differences in insulin and IGF-I receptor number alone could not explain these results. The two cell lines also respond differently to PDGF-AA. RN1a cells are relatively insensitive to stimulation by PDGF-AA and express fewer PDGF α receptors as shown by Northern blots and receptor-binding studies. We propose a unifying hypothesis in which the H+-ATPase activates a downstream element in the PDGF-AA signal transduction pathway that complements insulin and IGF-I signals, while leading to downregulation of the PDGF α receptor. © 1994 wiley-Liss, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041590319
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  • 4
    Electronic Resource
    Electronic Resource
    Growth and production characteristics of permeabilized Coleus blumei cells in immobilized fed-batch culture (1994)
    Springer
    Plant cell reports 13 (1994), S. 459-463 
    Details
    ISSN: 1432-203X
    Keywords: Coleus blumei ; Rosmarinic acid ; Immobilization ; DMSO ; Luffa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Permeabilized Coleus blumei cells were cultivated in an immobilized state to study the effect of dimethyl sulfoxide (DMSO) concentrations and growth regulators on cell growth and rosmarinic acid (RA) production characteristics. Luffa (the fibrous skeleton of mature fruit of Luffa cylindrica) was a good support matrix for cell immobilization because of its high void volume. Maximum cell loading capacity was 1.33 g dry cell weight (DCW)/g dry Luffa. The experiments were done in shake flasks with no free medium. The medium was supplied in a fed-batch mode to avoid the flotation of Luffa pieces. The sucrose in the medium was completely hydrolyzed to glucose and fructose without any sugar accumulation in the medium. The cell viability was slightly higher in the cells on top of the Luffa than those in the middle. Cell growth rate and rosmarinic acid (RA) production were approximately half that obtained in cell suspension cultures. Cell yield (g DCW/g glucose) was similar to that of cell suspension cultures. The absence of growth regulators did not promote an increase of RA production but did decrease the cell mass. The second step preconditioning with 0.5% DMSO did not improve the cell's adaptability to higher DMSO concentrations and the cell mass did not increase with 2.5% DMSO.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00231967
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  • 5
    Electronic Resource
    Electronic Resource
    Continuous limonin degradation by immobilizedRhodococcus fascians cells in K-carrageenan (1994)
    Springer
    Applied microbiology and biotechnology 41 (1994), S. 487-493 
    Details
    ISSN: 1432-0614
    Keywords: Limonin ; Debittering ; Immobilization ; Rhodococcus fascians ; Continuous Stirred Tank ; Reactor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Limonin can be effectively degraded byRhodococcus fascians cells. These bacteria can be entraped in κ-carrageenan, and used in a continuous stirred tank reactor to degrade limonin in a continuous process. The effects of temperature limonin concentration, dilution rate, and aeration on the reactor behaviour have been tested, and the results correlated with changes in limonin conversion, substrate degradation rate, and free and immobilized biomass. Results showed that the immobilized cells were able to debitter limonin-containing media and the immobilized biomass was quite stable throughout the operational conditions tested. A population of free biomass was present in the reactor, the quantity of which was dependent on dilution rate. The immobilized bacteria increased its limonin-degrading capability when the substrate concentration was increased. The aeration was not strictly necessary for limonin degradation. Additionally, the immobilized cells were active and stable for more than 2 months of continuous operation, and were able to recover their limonin-degrading capability when used intermittently. Finally, none of the main components of a juice was noticeably altered during limonin degradation, so the reactor response was good enough to consider its application.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01982541
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  • 6
    Electronic Resource
    Electronic Resource
    Continuous limonin degradation by immobilizedRhodococcus fascians cells in K-carrageenan (1994)
    Springer
    Applied microbiology and biotechnology 41 (1994), S. 487-493 
    Details
    ISSN: 1432-0614
    Keywords: Limonin ; Debittering ; Immobilization ; Rhodococcus fascians ; Continuous Stirred Tank ; Reactor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Limonin can be effectively degraded byRhodococcus fascians cells. These bacteria can be entraped in κ-carrageenan, and used in a continuous stirred tank reactor to degrade limonin in a continuous process. The effects of temperature limonin concentration, dilution rate, and aeration on the reactor behaviour have been tested, and the results correlated with changes in limonin conversion, substrate degradation rate, and free and immobilized biomass. Results showed that the immobilized cells were able to debitter limonin-containing media and the immobilized biomass was quite stable throughout the operational conditions tested. A population of free biomass was present in the reactor, the quantity of which was dependent on dilution rate. The immobilized bacteria increased its limonin-degrading capability when the substrate concentration was increased. The aeration was not strictly necessary for limonin degradation. Additionally, the immobilized cells were active and stable for more than 2 months of continuous operation, and were able to recover their limonin-degrading capability when used intermittently. Finally, none of the main components of a juice was noticeably altered during limonin degradation, so the reactor response was good enough to consider its application.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00939041
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  • 7
    Electronic Resource
    Electronic Resource
    Responses of salivary acinar cells to intracellular alkalinization (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 159 (1994), S. 457-467 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Responses of rat submandibular acini to intracellular alkalinization were investigated. Intracellular alkalinization was induced by addition of NH4Cl or methylamines, or by prepulse with Na butyrate. Only partial recovery occurred following Na butyrate prepulse or methylated amine addition, but full recovery was observed following addition of NH4Cl. The latter recovery was DIDS and dimethylamiloride-insensitive but was inhibited by bumetanide or high [K+] and stimulated in Na+ free buffer and by ouabain. Acetylcholine stimulated recovery from NH4Cl- or Na butyrate pre-pulse-induced alkalinization and reduced the extent of alkalinization induced by methylated amines. Acetylcholine-stimulated recovery from NH4Cl-induced alkalinization was mimicked by substance P or ionomycin and was partially Ca2+-dependent. This stimulated recovery was bumetanide-insensitive but was partially sensitive to charybdotoxin. Taken together, these data indicate that in unstimulated cells, recovery from alkalinization induced by NH4Cl occurs by bumetanide-sensitive transport of the NH4+ ion, that DIDS-inhibitable anion transport contributes little to this recovery, and that acetylcholine and other Ca2+-elevating agents accelerate recovery from NH4Cl-induced alkaline challenge by a mechanism insensitive to bumetanide, DIDS, ouabain, and dimethylamiloride but sensitive to extracellular Ca2+ and to charybdotoxin. Partial recovery from alkaline challenge can also occur in the absence of NH4+ ions, and acetylcholine also stimulates this mode of recovery. Together, these data suggest that these cells have little intrinsic ability to recover from intracellular alkalinization and that the NH4+ ion may be a surrogate for K+ in at least two ion transport pathways. © 1994 wiley-Liss, Inc.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041590310
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  • 8
    Electronic Resource
    Electronic Resource
    Acrosome reaction of boar spermatozoa in homologous in vitro fertilization (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 36 (1993), S. 84-88 
    Details
    ISSN: 1040-452X
    Keywords: Capacitation ; Triple stain technique ; Lectin ; Oocyte ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Ejaculated spermatozoa from four different boars were used to evaluate the acrosome reaction during in vitro fertilization with homologous ovulated oocytes. The acrosome reaction was assessed according to a peroxidase-labeling peanut agglutinin method and a triple-stain technique. An increase in the proportion of living sperm with reacted acrosomes was observed after preincubation and 2 hr of coincubation (P 〈 0.05). The percentage of true acrosome-reacted sperm remained reasonably constant throughout coincubation. In vitro penetration rates of oocytes varied among boars, but no relationship was found between fertilization rates of oocytes and maximum percentages of acrosome reacted living sperm. © 1993 Wiley-Liss, Inc.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1080360112
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  • 9
    Electronic Resource
    Electronic Resource
    Intracytoplasmic ciliary elements in epidermal cells of Syndesmis echinorum and Paravortex cardii (platyhelminthes, dalyellioida) (1992)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 213 (1992), S. 147-157 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Epidermal cells of Syndesmis echinorum and Paravortex cardii contain many intracytoplasmic ciliary components: clusters of centrioles disorganized and incomplete short axonemes composed of loosely organized microtubules of irregular lengths, fully formed axonemes though some with fewer than nine doublets, and ciliary rootlets. Furthermore, conspicuous dense granules are found in solitary groups in the cytoplasm. Clusters of dense granules are also closely associated with Golgi complexes and developing axonemal microtubules. Since the dense granules decrease in number as the axonemes increase, it is likely that the granules are involved in the formation of axonemal microtubules.Ciliary elements are especially abundant in epidermal cells of Paravortex cardii embryos, some of them resembling those previously described by several authors in differentiating ciliated cells engaged in centriologenesis and ciliogenesis. Attention has been focused on the relative proportion and position of these elements, as well as the different morphology and several assembling states that they exhibit in epidermal cells of adult S. echinorum and adults and embryos of P. cardii. A functional interpretation of some of the findings is given, which allows us to suggest a sequence of ciliogenetic events that occur in epidermal cells of both species. © 1992 Wiley-Liss, Inc.
    Additional Material: 18 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jmor.1052130202
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  • 10
    Electronic Resource
    Electronic Resource
    Choanocyte-like cells in the digestive system of the starfish Marthasterias glacialis (Echinodermata) (1991)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 208 (1991), S. 215-225 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Two types of choanocyte-like cells have been found in the digestive tract of the starfish. Type I choanocytes are in the lining epithelium of all organs of the digestive system. These are narrow, columnar cells strongly anchored basally and expanded apically into a protuberance projecting into the lumen. A prominent flagellum surrounded by microvilli projects from the center of this protuberance. Apical cytoplasm contains numerous mitochondria, secondary lysosomes, and multivesicular bodies. A distinctive characteristic of these cells is a filament bundle that traverses the length of the cell from its region of attachment on the rootlet of the flagellar basal body to its terminus on the basal plasma membrane. Between the attenuated basal ends of type I cells are the nerve fibers of an intraepithelial nerve plexus. Thickness of the plexus is correlated with the quantity of type I cells in the epithelium.Type II choanocytes are in the cuboidal coelomic epithelium that forms the outer layer of digestive tract organs. These cells are smaller than those of type I, and they have an apical collar surmounted by a ring of 13 microvilli. Within the collar is a cup-shaped depression with a central flagellum. Coated vesicles, secondary lysosomes, and phagocytic infoldings are observed in and near the collar cytoplasm. Filament bundles similar to those in type I choanocytes are also observed in coelomic epithelial cells that are sufficiently tall. Injection of peroxidase into the stomach and ferritin into the coelom results in phagocytic uptake of these macromolecules by type I and type II choanocytes, respectively.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jmor.1052080207
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