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  • 1
    ISSN: 1476-5535
    Keywords: Keywords: biotin production; E. coli bio operon; Agrobacterium/Rhizobium HK4; limiting growth conditions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: E. coli biotin (bio) operon was modified to improve biotin production by host cells: (a) the divergently transcribed wild-type bio operon was re-organized into one transcriptional unit; (b) the wild-type bio promoter was replaced with a strong artificial (tac) promoter; (c) a potential stem loop structure between bioD and bioA was removed; and (d) the wild-type bioB ribosomal binding site (RBS) was replaced with an artificial RBS that resulted in improved bioB expression. The effects of the modifications on the bio operon were studied in E. coli by measuring biotin and dethiobiotin production, and bio gene expression with mini-cells and two-dimensional polyacrylamide gel electrophoresis. The modified E. coli bio operon was introduced into a broad host-range plasmid and used to transform Agrobacterium/Rhizobium HK4, which then produced 110 mg L−1 of biotin in a 2-L fermenter, growing on a defined medium with diaminononanoic acid as the starting material. Biotin production was not growth-phase dependent in this strain, and the rate of production remained high under limiting (maintenance) and zero growth conditions.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0851
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Active specific immunotherapy was performed in a phase I study in 20 colorectal cancer patients after surgical resection of the tumor. An autologous tumor cell vaccine surface modified by Newcastle disease virus (NDV) was used, which showed the following characteristics. After mechanical and enzymatic dissociation of the tumor tissue an average of 5 × 107 cells/g tissue was obtained. According to trypan blue dye exclusion assay the average viability was 72%. Following irradiation (200 Gy) the inactivation of proliferative activity of the cells could be demonstrated by the absence of incorporation of3H-labelled thymidine. The cells were, however, still metabolically active as shown by the incorporation of [3H]-uridine and a mixture of3H-labelled amino acids. Epithelium-specific antigens (detected by mAb HEA125) were expressed on more than 75% cells of the cell suspension indicating a high amount of (epithelium-derived) tumor cells. In order to increase the immunogenicity of the tumor cells the suspended cells were infected by the nonlytic, apathogenic Ulster strain of NDV. The successful modification of tumor cells with NDV could be shown by electron microscopy. Three weeks postoperatively cells were thawed, virus-modified, and inoculated intradermally in the upper thigh. Several cell and virus concentrations were tested in each patient. As control, tumor cells without NDV, NDV alone and normal colon mucosa were used. The number of tumor cells ranged from 2 × 106 up to 2 × 107 cells and NDV concentrations from 4 to 64 hemagglutination units (HU) were tested. Sixteen patients responded with a delayed-type hypersensitivity (DTH) skin reaction to the vaccine. The best DTH reaction, measured 24 h following vaccination, was obtained using a vaccine consisting of 1 × 107 tumor cells and 32 HU NDV (median induration of 8 mm). Response to NDV alone was seen in 2 patients only (median induration of 3 mm); 12 patients responded to tumor cells (1 × 107) alone (median induration of 4 mm). Of 10 patients tested with normal colorectal mucosa, 4 responded with a median induration of 3.5 mm. DTH responses to the vaccine of 1 × 107 tumor cells and 32 HU NDV increased throughout the repeated vaccinations to a median induration of 9.5 mm at the end of the therapy. No severe side-effects in the course of the immunotherapy, except for mild fever in 4/20 patients, were observed. The results of our phase I study show that this type of autologous colorectal tumor cell vaccine is ready for a large clinical trial to prove its efficacy.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1435-2451
    Keywords: Esophageal carcinoma ; Operative therapy ; Preoperative (neoadjuvant) chemotherapy ; Oesophagus-carcinom ; Operative Therapie ; Preoperative Therapie
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung 42 Patienten mit nicht fernmetastasiertem Plattenepithel-Carcinom des Oesophagus wurden präoperativ cytostatisch vorbehandelt (Kelsen-Schema). Bei 18 Patienten kam es zu einer partiellen, bei 2 Patienten zu einer vollständigen Tumorremission. Von 40 Patienten, die sich einem operativen Eingriff unterzogen, waren 4 inoperabel. Bei 14 Patienten konnte der Tumor palliativ und bei 22 potentiell kurativ entfernt werden. Postoperativ verstarben 4 der 36 resezierten Patienten. Eine Anastomoseninsuffizienz trat bei 5 Patienten und schwere kardio-pulmonale Komplikationen bei 4 Patienten postoperativ auf. Die Nebenwirkungen der präoperativen Behandlung waren akzeptabel und werden durch das Ergebnis der Behandlung mit einer hohen Resektabilitätsrate und einer Verbesserung der Überlebenszeit von Patienten, die auf die präoperative Chemotherapie ansprechen, gerechtfertigt.
    Notes: Summary 42 patients with localized squamous cell carcinoma of the esophagus were treated according to a phase II study with cisplatin, vindesine and bleomycin (KelsenSchema) prior to surgery. In 18 of these patients partial remission was achieved and in two cases complete remission. Of the 40 patients which were presented to surgical treatment, 4 were inoperable, in 22 cases the tumor was removed in a potentially curative manner and in 14 patients a palliative resection was performed. The postoperative mortality was 4 of 36 resected patients. Anastomotic leakage was found in 5 patients and severe cardiopulmonary complications in 4 patients. The side effects of the preoperative treatment were acceptable and seem justified by the high resectability rate and the significant improvement in survival of patients who responded to chemotherapy.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Parasitology research 37 (1971), S. 288-302 
    ISSN: 1432-1955
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Adulte Dicrocoelium dendriticum enthielten 8,6 mg Protein, 7,5 mg Kohlenhydrate (davon 5,7 mg Glykogen), 2,9 mg Lipide und 1,2 mg Asche pro 100 mg Frischgewicht. Die Atmung der intakten Parasiten wurde polarographisch und manometrisch gemessen. Die durchschnittliche Rate des endogenen Sauerstoffverbrauches betrug bei manometrischen Messungen 16,2 μl O2/mg N/Std. Dies entspricht einer Rate von 1,2 μl O2/mg Trockensubstanz/Std. Die Sauerstoffaufnahme der Parasiten war von der Sauerstoffspannung abhängig. Im Bereich zwischen 21 Vol.-% O2 und 5% blieb die Rate annähernd konstant, sie war jedoch bei 100% deutlich erhöht und bei 2% signifikant vermindert. In manometrischen Versuchen bewirkte Glycerol (Endkonzentrationen 10 mM und 20 mM) eine deutliche Steigerung des Sauerstoffverbrauches, während Glucose, Galaktose und Fructose ohne Einfluß auf die Atmung blieben. In den polarographischen Untersuchungen war dagegen kein Effekt exogener Substrate zu erkennen. Einige der als Inhibitoren der Atmungskette in Säugetier-Mitochondrien bekannten Substanzen (2-Thenoyltrifluoraceton, 8-Hydroxychinolin, Salicylaldoxim, KCN und Antimycin A) verursachten eine deutliche Hemmung des Sauerstoffverbrauches. Die Rate des Sauerstoffverbrauches wurde durch 2,4-Dinitrophenol stimuliert, vermutlich infolge der Entkopplung der oxydativen Phosphorylierung. Glucose und Glycerol, einzeln in verschiedenen Konzentrationen zu zuckerfreier Tyrode-Lösung zugefügt, wurden von D. dendriticum unter aeroben oder anaeroben Bedingungen aufgenommen. Die Absorptionsraten waren mit den Substratkonzentrationen im Medium positiv korreliert. Aerob war die Substrataufnahme etwas höher als anaerob. Die Raten der Lactatausscheidung waren unter aeroben und anaeroben Bedingungen etwa gleich groß und unabhängig von den Substratkonzentrationen.
    Notes: Summary Adult Dicrocoelium dendriticum contained 8.6 mg protein, 7.5 mg carbohydrates (including 5.7 mg glycogen), 2.9 mg lipids and 1.2 mg ash per 100 mg fresh weight. The respiration of intact parasites was studied by means of polarographic and manometric techniques. The average rate of the endogenous oxygen uptake determined manometrically was 16.2 μl O2/mg N/h. This corresponds with a value of 1.2 μl O2/mg dry substance/h. The respiration of the parasites responded to changes of the oxygen tension. The rate of oxygen consumption was nearly constant at oxygen tensions between 21 and 5 vol.-%, but it was significantly increased at 100% and deminished at 2%. Added glycerol (final concentrations of 10 mM and 20 mM) increased the rate of oxygen uptake in manometric experiments, whereas glucose, galactose and fructose produced no stimulatory effect. No influence of exogenous substrates was detectable in experiments using the polarographic technique. Some of the inhibitors of the respiratory chain in mitochondria of mammals (2-thenoyltrifluoroacetone, 8-hydroxyquinoline, salicylaldoxime, KCN, antimycin A) added to the media distinctly inhibited the oxygen consumption of D. dendriticum. The rate of respiration was increased by 2,4-dinitrophenol, probably due to the uncoupling of oxydative phosphorylation. Glucose or glycerol added separately to sugar-free Tyrode's solution at various concentrations was absorbed by D. dendriticum during aerobic and anaerobic conditions. The rates of absorption were positively correlated with the substrate concentrations in the medium. Glycerol was absorbed in larger amounts than glucose. The absorption of substrate was slightly higher under aerobic than under anaerobic conditions. The rates of lactate excretion were similar under aerobic and anaerobic conditions and independent of substrate concentrations.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    New York : Wiley-Blackwell
    Journal of Polymer Science: Polymer Letters Edition 20 (1982), S. 233-240 
    ISSN: 0360-6384
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
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