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  • 1
    Electronic Resource
    Electronic Resource
    Copenhagen : International Union of Crystallography (IUCr)
    Acta crystallographica 56 (2000), S. 200-202 
    ISSN: 1399-0047
    Source: Crystallography Journals Online : IUCR Backfile Archive 1948-2001
    Topics: Chemistry and Pharmacology , Geosciences , Physics
    Notes: A recombinant α-amylase from Bacillus stearothermophilus was found to be produced as several isoforms arising from different N-terminal processing. Some of those isoforms were purified to homogeneity and crystallized at 293 K using the hanging-drop vapour-diffusion method under the following conditions: 35 mM sodium acetate (pH 4.6), 6.25%(v/v) 2-propanol, in the presence of 1.23%(w/v) acarbose (a pseudo-oligosaccharide inhibitor) in the drop. The crystals diffracted beyond 2.0 Å resolution using synchrotron radiation at the Photon Factory, Tsukuba. They belong to the monoclinic space group P21, with unit-cell parameters a = 53.7 (2), b = 92.9 (4), c = 53.2 (2) Å, β = 109.4 (1)°.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Applied microbiology and biotechnology 20 (1984), S. 371-377 
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The basic problems of applying solvent extraction to ethanol fermentation were investigated. The selection of solvents was based on the selectivity ratio, which was expressed as the ratio of the ethanol distribution coefficient to the water distribution coefficient. Solvents with high selectivity ratios of more than 50 were found mainly among the alcohols and esters. However, most of these solvents were toxic to ethanol-producing microorganisms. We tried to make a barrier to solvent molecules beneath the surface of gel beads immobilizing the cells as a protection against solvent toxicity. Porapack Q was found to be an effective barrier, and the ethanol production rate of immobilized cells protected with Porapack Q did not change event after the production of eight batches in medium saturated with sec-octanol, which was the most toxic solvent used in our experiments.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Journal of artificial organs 2 (1999), S. 170-175 
    ISSN: 1619-0904
    Keywords: Remove immune complex ; Glomerular epithelial cell ; CR1 expression ; DNA/anti-DNA antibody ; Phagocytosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Technology
    Notes: Abstract Patients with systemic lupus erythematosus (SLE), an autoimmune disease caused by excessive amounts of immune complex in the serum, have a loss of complement receptor type 1 (CR1), a receptor of complements C3b and C4b, on glomerular epithelial cells. In order to calrify the biological function of CR1 on renal glomeruli and the correlation with clearance of immune complex, a full length of human tonsil CR1 cDNA was transfected to normal rat glomerular epithelial cells (SGE1) to enable CR1 expression in the long term (more than 1 year). As compared with the non-CR1-expressing cells, the CR1-expressing cells showed a higher binding effect to immune complex in 60 min, and the binding effect was mediated by complement. Blocking of the binding effect by anti-CR1 antibody indicates that CR1 plays an important role in removal of immune complex. Results from microscopy showing that the constructed cell has an enhanced phagocytic ability in the presence of complement suggest that the mechanism of removing immune complex by CR1-expressing cells has three steps: opsonization of immune complex by complement, the ligand-receptor interaction between opsonized complex and CR1 on the cell, and phagocytosis of complex by CR1-expressing cells.
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  • 4
    ISSN: 1573-0778
    Keywords: trehalose lipid ; U937 ; monocytic differentiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A novel type of succinoyl trehalose lipid (STL-1) prepared from n-hexadecane-culture ofRhodococcus erythropolis SD-74 markedly inhibited the growth of a human monocytoid leukemic cell line, U937, and induced its morphological alteration along a monocyte-macrophage lineage. STL-1 markedly increased differentiation-associated characteristics in macrophage, such as nitroblue tetrazolium reducing ability, appearance of Fc receptor, phagocytic activities in U937. Furthermore, U937 cells, which were activated with STL-1 exhibited cytotoxic activity against human lung carcinoma cell line A549. However, STL-1 did not affect growth of a normal human fetal lung cell line TIG-1. The individual components of STL-1, neither sugar moiety nor fatty acids in the free form, were effective at inducing the differentiation of U937 cell. From these results, we concluded that STL-1 has low cytotoxicity against normal human cells and the ester molecule itself is responsible for the activity of inducing differentiation of human monocytoid leukemic cell line U937 into monocyte-macrophage which results in the stimulation of the production of some cytotoxic substances.
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  • 5
    ISSN: 1573-0778
    Keywords: anti-DNA-antibody ; complement receptor ; immune complex ; NZB/W mice ; SLE
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract In vitro preparation of DNA/anti-DNA immune complex using monoclonal antibody and low molecular weight homogeneous DNA was described and the characteristics of the immune complex were identified. The immune complex was formed by the monoclonal antibody with DNA effectively at high ratios of antibody/DNA. The activation and binding ability of the immune complex to the complement was considerably high, whereas the capacity to bind red blood cells via C3b complement component receptor was shown relatively low. The assay of sucrose density gradient ultracentrifugation indicated that the sedimentation coefficient of the immune complex was between 10S and 23S. Studies of organ distribution and uptake of IC demonstrated a particular affinity to the kidney tissue. The significance of these results with respect to the latent pathogenicity of the immune complex was discussed.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Cytotechnology 7 (1991), S. 103-112 
    ISSN: 1573-0778
    Keywords: adaptation ; ammonia ; hybridoma ; continuous culture ; serum-free medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Using two mouse-mouse hybridoma cell lines, the response to ammonia step and serial changes was investigated in batch and continuous cultures with serum-free medium. The inhibitory effect of ammonia on cell growth depended on the cultivation mode, and differed markedly between cell lines. The cell line, 4C10B6 producing IgG monoclonal antibody against Pseudomonas, showed a high adaptation ability to ammonia. The 4C10B6 cells could grow under ammonia concentration as high as 21 mmol/l NH4Cl with a viability of 80% in the continuous culture with serial increase in ammonia concentration. Whereas, in the batch culture with ammonia step change the cell growth completely ceased at 12 mmol/l NH4Cl. The other cell line, TO-405 producing IgG monoclonal antibody against hepatitis B surface antigen, could not adapt to ammonia, and the cell growth did not occur at 9 mmol/l NH4Cl even under the ammonia serial change.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Cytotechnology 18 (1995), S. 35-50 
    ISSN: 1573-0778
    Keywords: ammonium removal ; cross-flow filtration ; monoclonal antibody ; membrane fouling ; perfusion culture ; zeolite ; hybridoma cells ; defined medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Production of monoclonal antibody against hepatitis B surface antigen was carried out by perfusion culture coupled with a selective removal system for ammonium ion. The removal system is composed of three sub-systems namely, cell separation by cross-flow ceramic filter, dialysis by hollow fiber module and ion-exchange by zeolite A-3 packed bed column. The ammonium ion concentration in the culture broth was effectively maintained below the inhibitory level, and the viable cell density reached 2.5×107 cells ml−1 which was three times that of conventional perfusion cultures. The monoclonal antibody accumulated to a concentration as high as 26.3×105 mIU−1. This is already almost half of the amount producedin vivo. The numerical investigation of the ammonium ion removal system showed the possibility to improve much more the performance of this perfusion cultivation system.
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  • 8
    ISSN: 1573-0778
    Keywords: ammonia removal ; hybridoma ; HBs monoclonal antibody ; zeolite
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Serum-free perfusion cultures of hybridoma TO-405 cells were carried out in spinner flasks coupled with zeolite A-3 packed beads. Ammonia was selectively removed from the culture broth by passing cell free permeate from ceramic cross flow filtration, through the zeolite packed bed. Ammonia concentration in the culture broth was effectively maintained between 1 to 4 mmol/l which was below the inhibitory concentration for cell growth. Maximum cell density levels of 107 cells/ml as well as improved percentage cell viability higher than in serum-supplemented cultures were feasible in this system. The possible effects of shear stress, generated by variation of the flow rates of the broth through the ceramic filter module, on the growth of the hybridoma cells were investigated. Backwashing, by reversing the direction of the permeate, was found necessary to prolong the life of the filter. Variation of the flow rates of the broth through the ceramic module between 0.29 m/s to 0.59 m/s did not cause immediate cell damage but growth was repressed at the higher flow rate. This study also showed that glutamine appears to be one of the factors limiting the growth of the hybridoma cells.
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  • 9
    ISSN: 1573-0778
    Keywords: adoptive immunotherapy ; cytotoxic T lymphocyte ; formalin-fixed tumor cell
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Human tumor specific cytotoxic T lymphocytes (CTL) were expanded on formalin-fixed autologous target tumor cells derived from glioblastoma multiforme. Growth response of the CTL restimulated with the fixed target cells was larger than those with live target cells. The results suggest that formalin-fixed tumor cells will be stable sources of tumor antigen for efficient autologous CTL expansion and be useful for adoptive immunotherapy of tumors.
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  • 10
    ISSN: 1573-0778
    Keywords: biosurfactants ; differentiation ; microbial extracellular glycolipids ; PC12
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The effects of several kinds of microbial extracellular glycolipids on neurite initiation in PC12 cells were examined. Addition of mannosylerythritol lipid-A (MEL-A), MEL-B, and sophorose lipid (SL) to PC12 cells caused significant neurite outgrowth. Other glycolipids, such as polyol lipid (PL), rhamnose lipid (RL), succinoyl trehalose lipid-A (STL-A) and STL-B caused no neurite-initiation. MEL-A increased acetylcholine esterase (AChE) activity to an extent similar to nerve growth factor (NGF). However, MEL-A induced one or two long neurites from the cell body, while NGF induced many neurites. In addition, MEL-A-induced differentiation was transient, and after 48 h, percentage of cells with neurites started to decrease in contrast to neurons induced by NGF, which occurred in a time-dependent manner. MEL-A could induce neurite outgrowth after treatment of PC12 cells with an anti-NGF receptor antibody that obstructed NGF action. These results indicate that MEL-A and NGF induce differentiation of PC12 cells through different mechanisms.
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