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H G Miltenburger

Publications and source records attributed to H G Miltenburger.

At least 19 recordsLinked to original sources
PubMed2000

In vitro micronucleus assay with Chinese hamster V79 cells - results of a collaborative study with in situ exposure to 26 chemical substances.

A collaborative study with 10 participating laboratories was conducted to evaluate a test protocol for the performance of the in vitro micronucleus (MN) test using the V79 cell line with one treatment and one sampling time only. A total of 26 coded substances were tested in this study for MN-inducing properties. Three substances were tested by all 10 laboratories and 23 substances were tested by three or four laboratories in parallel. Six aneugenic, 7 clastogenic and 6 non-genotoxic chemicals were uniformly recognised as such by all laboratories. Three chemicals were tested uniformly negative by three laboratories although also clastogenic properties have been reported for these substances. Another set of three clastogenic substances showed inconsistent results and one non-clastogenic substance was found to be positive by one out of three laboratories. Within the study, the applicability of the determination of a proliferation index (PI) as an internal cytotoxicity parameter in comparison with the determination of the mitotic index (MI) was also evaluated. Both parameters were found to be useful for the interpretation of the MN test result with regard to the control of cell cycle kinetics and the mode of action for MN induction. The MN test in vitro was found to be easy to perform and its results were mainly in accordance with results from chromosomal aberration tests in vitro.

Animals↗
PubMed1994

Cooperative effects in vitro on fibroblast and keratinocyte functions related to wound healing by transforming growth factor-beta and a low molecular weight fraction from hemolyzed blood.

Blood or its constituents, respectively, contain a.o. substances such as TGF-beta (transforming growth factor-beta), PDGF (platelet-derived growth factor) and other factors, which beneficially influence wound healing. Patients with perivascular disturbances and consequently with inadequate supply of the affected tissue cells often suffer from poor healing of dermal wounds. Here, various cellular functions in situations of poor supply and related to wound healing, such as proliferation, colony formation, and migration of fibroblasts, and of monolayer formation by keratinocytes were emulated in vitro by supplementing cultures with reduced amounts of serum. Computer-aided image analysis allowed to quantify the cellular reactions under normal and serum-deprived medium conditions and under the influence of a low molecular weight fraction manufactured by dialysis of hemolyzed calf blood (HD, Solcoseryl) and of TGF-beta. Both preparations are in use for the treatment of poorly healing wounds. While HD preferentially normalized the reduced viability of fibroblasts, the keratinocyte activity was enhanced by TGF-beta. Restoration of fibroblast and keratinocyte functions proved most effective when combining HD with TGF-beta.

Actihaemyl↗
PubMed1993

Biotechnological aspects of the production of human pro-kallikrein using the AcNPV-baculovirus-expression system.

We have investigated large scale production processes (up to 2 liters) of recombinant proteins using the baculovirus expression system in order to optimize the product yields. Experiments using cell lines of Spodoptera frugiperda (Sf9) and Mamestra brassicae (IZD-Mb0503) were performed to show the different production capacities of the cell lines. The influence of the infection at different cell densities is described. Beyond that, TC100-, IPL41- and serum-free IPL41-medium were compared to demonstrate their different capabilities of supporting cell growth and protein expression. Additionally, the inhibitory effect of FCS on the protease activity of kallikrein, which is produced in its zymogenic form, is discussed Improved production parameters are described, which enabled us to produce up to 8000 units of activated pro-kallikrein within 14 days using perfusion cultivation.

Animals↗
PubMed1993

The genotoxicity status of senna.

Genotoxicity tests were performed by several laboratories with the drug fructus sennae, senna extract, sennosides, rhein and aloe-emodin. The drug fructus sennae, the sennosides and rhein did not increase mutation frequencies in the following test systems: bacterial systems (Salmonella reverse mutation test and/or Escherichia coli forward mutation test); mammalian cell cultures [hypoxanthine guanine phosphoribosyl transferase (HGPRT) test; mouse lymphoma test; chromosome aberration test with Chinese hamster ovary cells]; bone marrow (micronucleus test; chromosome aberration test); melanoblast cells (mouse spot test) of rodents. With aloe-emodin mutagenic effects were observed only in vitro in the chromosome aberration test with CHO cells and in the Salmonella reverse mutation test (frameshift mutations in strains TA 1537, TA 1538 and TA 98). In the in vitro gene mutation test with V79 cells (HGPRT test) no mutagenic potential of aloe-emodin was observed. In in vivo studies [micronucleus test with bone marrow cells of NMRI mice, chromosome aberration test with bone marrow cells of Wistar rats, mouse spot test (crossing DBA/2J x NMRI) no indication for a mutagenic activity of aloe-emodin was found. The relevance of the absence of a mutagenic potential in in vivo test systems was strengthened by the fact that aloe-emodin could be found in the blood serum after oral administration. Additional information on the interaction of aloe-emodin with DNA was obtained from an ex vivo unscheduled DNA synthesis test performed with hepatocytes of male Wistar rats: aloe-emodin did not induce unscheduled DNA synthesis as expression of DNA damage.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Construction and evaluation of a cea-lacZ gene fusion for the detection of environmental mutagens and carcinogens.

The cea-kil operon of the ColE1 plasmid is negatively regulated by the LexA-repressor and therefore, it is under the control of SOS regulation. We constructed a gene fusion between the cea and lacZ genes. Expression of the translational fusion can be easily detected by monitoring the levels of beta-galactosidase. Since the whole detection system is plasmid-based, it can be used in both Escherichia coli and Salmonella typhimurium strains. The SOS-function-inducing activities of 14 chemical mutagens were investigated in E. coli K12 and in two S. typhimurium Ames-strains and compared with results obtained by the SOS-chromotest and by the Umu-test. To correct for the inhibitory effects of test chemicals on mRNA and/or protein synthesis, the level of the constitutive chloramphenicol acetyl transferase was assayed in parallel.

Bacillus subtilis↗
PubMed1991

Report of a comparative study of DNA damage and repair assays in primary rat hepatocytes with five coded chemicals.

We report the results of a collaborative study for the detection of chemical-induced DNA damage in primary cultures of rat hepatocytes. The methods include the detection of unscheduled DNA synthesis (UDS) with either autoradiography (5 laboratories) or liquid scintillation counting (2 laboratories) and the assessment of DNA single-strand breaks with the alkaline elution assay (1 laboratory). Interlaboratory standardization was omitted in order to prove the agreement of the assays under routine conditions. Five coded chemicals were tested. For 4 chemicals (2-acetylaminofluorene, thiourea, glycerine and potassium chloride) the UDS data were consistent in all laboratories, thus indicating a high consensus of the test systems applied in the different laboratories. Those 3 chemicals that were not expected to elicit genotoxic activity (thiourea, glycerine, and potassium chloride) yielded negative results in all laboratories. 2-Acetylaminofluorene, a known DNA-damaging agent in hepatocytes, gave strongly positive responses in all laboratories. In contrast, N-nitrosodiphenylamine led to equivocal responses.

2-Acetylaminofluorene↗
PubMed1991

Inter-laboratory ring trial of in vitro DNA repair tests using rat hepatocytes: further testing and conclusive remarks.

The results of the extension of a collaborative study for the detection of chemical-induced DNA damage in rat hepatocytes in vitro are presented in this report. Three coded compounds, i.e. 1,4-butanediol dimethanesulphonate, hydrazine sulphate and sodium dichromate, were tested for DNA repair synthesis by seven different laboratories, either using autoradiographic procedures or the liquid scintillation counting technique. Inter-laboratory standardization was intentionally not requested in order to investigate the validity of each study design under routine conditions. 1,4-Butanediol dimethanesulphonate was clearly positive in most laboratories; sodium dichromate was generally positive, while the results on hydrazine sulphate were contradictory.

Animals↗
PubMed1990

Multistage production of Autographa californica nuclear polyhedrosis virus in insect cell cultures.

The aim of our study was to establish an efficient system for the in vitro production of the insect pathogenic Autographa californica nuclear polyhedrosis virus in a Spodoptera frugiperda cell line. We optimized cultivation conditions for cell proliferation as well as for virus replication in a 1.5 litre stirred tank bioreactor. Cell and virus propagation were found to be optimal at a constant oxygen tension of 40%. In order to provide sufficient nutrients during virus synthesis filtration and perfusion devices were connected to the bioreactor. A virus production procedure in a repeated batch mode by using a two stage bioreactor system is described. Stage I was optimized for cell production and stage II for virus production.

Animals↗
PubMed1990

Cell cycle kinetics of insect cell cultures compared to mammalian cell cultures.

Cell cycle kinetic of lepidopteran cell lines Sf9 (Spodoptera frugiperda) and IZDMb0503 (Mamestra brassicae) were investigated and compared to mammalian cell cycle distributions. The resting phase (G0) of mammalian cells is characterized by a 2c-DNA content whereas G0-phase of insect cell lines is characterized by a 4c-DNA content. Flow cytometric data in combination with growth curves of partially synchronized and asynchronously growing cells proved the existence of this phenomenon. Kinetics of cells labeled by the thymidine analog on 5-bromo-2'-deoxyuridine supported these results, which now render the possibility of applying cell cycle analysis in fermentation technology of insect cells.

Animals↗
PubMed1990

[Genotoxicity of a standardized Hypericum extract].

St. John's wort (Hypercum perforatum) contains hypericin and hypericin-like substances as well as flavonoids, of which particularly Quercetin has generated a wide-spread controversial discussion with respect to mutagenic action. The genotoxicity of a standardized aqueous ethanolic Hypericum extract (Hypericum extract Steigerwald, Psychotonin M) was verified in different in-vivo and in-vitro testsystems with mammalian cells. The in-vitro investigations were performed with the HGPRT (hypoxanthine guanidine phosphoribosyl transferase)-test, UDS (unscheduled DNA synthesis)-test and with the cell transformation test using Syrian hamster embryo cells. Both the in-vitro tests as well as the in-vivo tests--fur spot test of the mouse and the chromosome aberration test with the bone marrow cells of the chinese hamster--were negative, giving completely no indication of a mutagenic potential of Hypericum extract. These investigations lend support to the view that results from bacterial short-term tests are of very limited transferability to human.

Animals↗
PubMed1989

Flow-cytometric cell-cycle analysis of Chinese hamster cells following exposure to cytotoxicants.

Chinese hamster cell-cycle kinetics were studied following exposure to a cytotoxic agent. Different parameters like dependence on concentration and preparation interval were measured by flow cytometry. The results were compared with data from established methods. DNA histograms showed the cell-cycle effect of EMS (ethyl methanesulfonate) at different time intervals after treatment. The principle of quenching the fluorescence of Hoechst 33258 by staining 5-bromodeoxyuridine (BrdU) substituted DNA was applied and supports these results. Comparison to chromosome-aberration studies demonstrates the suitability of this method to screen quickly for adequate dosing of a cytotoxic substance and also gives information on the appropriate preparation interval.

Animals↗
PubMed1987

S-phase cell detection with a monoclonal antibody.

The use of a newly developed monoclonal antibody against Bromodeoxyuridine is demonstrated in a study with V79 Chinese hamster cells. It can replace radioactive material in cell cycle analysis studies in the way that BrdU incorporated into the DNA during S-phase is detected by immunofluorescence. By this technique the proportion of actively synthesizing S-phase cells can be determined. The proportion of such cells is a good measure with regard to the quality of a culture and/or medium in question. The method is fast and offers within 3-5 hours the possibility to analyse growth conditions in monolayers and suspension cultures.

Animals↗
PubMed1987

Cell cycle analysis by flow cytometry in an antibody secreting cell line.

This presentation reports on the routine use of flow cytometric techniques in suspension cultures. As a model system an antibody secreting mouse hybridoma cell line was used to monitor antibody production (a Bromodeoxyuridine specific antibody) and proliferative status as obtained by flow cytometry (FCM) and cell counting.

Animals↗
PubMed1985

Heavy-ion effects on mammalian cells: inactivation measurements with different cell lines.

In track segment experiments, the inactivation of different mammalian cells by heavy charged particles between helium and uranium in the energy range between 1 and 1000 MeV/u has been measured at the heavy ion accelerator Unilac, Darmstadt, the Tandem Van de Graaf, Heidelberg, and the Bevalac, Berkeley. The inactivation cross sections calculated from the final slope of the dose-effect curves are given as a function of the particle energy and the linear energy transfer.

Animals↗
PubMed1985

Qualitative and quantitative aspects of animal cell in vitro systems.

The evaluation of sublethal cellular damage is shown to be important for the performance of experiments and bioproduction with animal cells. Detailed information on the fate of single cells in colony pedigrees from untreated and irradiated mammalian cells was obtained in vitro with cinematography recordings. It can be demonstrated that in generations two, three, and four the cells are more sensitive because of higher death rates and increased generation times induced by the handling of the cells during passaging or by the damaging agent. It is obvious that chemical agents can act in the same way. Also poor medium quality induces similar effects. Growth curves of single colony cell populations render much better information on the proliferation of a cell population than the determination of cells per ml (= particle counting). Examples are given for such growth curves and for colony size distribution curves based on individual cell countings. The advantages of the use of flow-cytofluorometry for the control of cell cycle phases are shown. As an example for the relevance of qualitative and quantitative data on cell substrate and medium conditions for bioproduction the in vitro mass replication of an insect pathogenic virus (baculovirus) is presented. Baculoviruses are potentially important biological pesticides which can replace chemical insecticides partially.

Animals↗
PubMed1984

HZE effects on mammalian cells.

In track segment experiments cell survival and chromosome aberrations of mammalian cells have been measured for various heavy ion beams between helium and uranium in the energy range between 0.5 and 960 MeV/u, corresponding to a velocity range of 0.03 to 0.87 C, and an LET spectrum from 10 to 15 000 keV/micrometers. At low LET, the cross section (sigma) for cell killing increases with increasing LET and shows a common curve for all ions regardless of the atomic number. This indicates that in this region the track structure of the different ions is of only a minor influence, and it is rather the total energy transfer, which is important for cell killing. At higher LET values, deviations from a common sigma-LET curve can be observed which indicate a saturation effect. The saturation of the lighter ions occurs at lower LET values than for the heavier ions. These findings are also confirmed by the chromosome data, where the efficiency for the induction of chromosomal aberrations for high LET particles depends on the track structure and is nearly independent of LET. In the heavier beams (Z > or = 10) individual particles cause multiple chromosome breaks in mitotic cells.

Animals↗