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L Müller

Publications and source records attributed to L Müller.

At least 37 records · Page 2Linked to original sources
PubMed2001

Dendritic biomimicry: microenvironmental hydrogen-bonding effects on tryptophan fluorescence.

Two series of dendritically modified tryptophan derivatives have been synthesised and their emission spectra measured in a range of different solvents. This paper presents the syntheses of these novel dendritic structures and discusses their emission spectra in terms of both solvent and dendritic effects. In the first series of dendrimers, the NH group of the indole ring is available for hydrogen bonding, whilst in the second series, the indole NH group has been converted to NMe. Direct comparison of the emission wavelengths of analogous NH and NMe derivatives indicates the importance of the Kamlet-Taft solvent beta3 parameter, which reflects the ability of the solvent to accept a hydrogen bond from the NH group, an effect not possible for the NMe series of dendrimers. For the NH dendrimers, the attachment of a dendritic shell to the tryptophan subunit leads to a red shift in emission wavelength. This dendritic effect only operates in non-hydrogen-bonding solvents. For the NMe dendrimers, however, the attachment of a dendritic shell has no effect on the emission spectra of the indole ring. This proves the importance of hydrogen bonding between the branched shell and the indole NH group in causing the dendritic effect. This is the first time a dendritic effect has been unambiguously assigned to individual hydrogen-bonding interactions and indicates that such intramolecular interactions are important in dendrimers, just as they are in proteins. Furthermore, this paper sheds light on the use of tryptophan residues as a probe of the microenvironment within proteins--in particular, it stresses the importance of hydrogen bonds formed by the indole NH group.

Fluorescence↗
PubMed2001

Molecular approach to the characterisation of fungal communities: methods for DNA extraction, PCR amplification and DGGE analysis of painted art objects.

A protocol for efficient extraction of fungal DNA from micromycetes colonising painted art objects was developed. Polymerase chain reaction (PCR) inhibitors were successfully removed by a combined application of a Chelex-100 adsorption resin and a Geneclean Kit for Ancient DNA. Universal fungal primers for PCR amplification of 28S rDNA (U1 and U2) were tested for their applicability in denaturing gradient gel electrophoresis (DGGE) analysis of fungal communities. Artificially produced mortar samples inoculated with fungal pure cultures isolated from mural paintings were used as model objects for DNA extractions and DGGE analysis. Good resolution in DGGE was achieved using 260-bp rDNA fragments amplified with U1/DGGE and U2 primers directly from model communities.

Biofilms↗
PubMed2001

Levels of metals and organic substances in blood and urine of workers at a new hazardous waste incinerator.

OBJECTIVE: To assess baseline concentrations of a number of metals and organic compounds in blood and urine of 28 workers employed at a new hazardous waste incinerator (HWI), before operation of the plant. METHODS: Plasma analyses of hexachlorobenzene (HCB), polychlorinated biphenyls (PCB 28, 52, 101, 138, 153 and 180), benzene, toluene, ethylbenzene, m-xylene, and polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) were carried out. The urinary levels of 2,4- and 2,5-dichlorophenol (DCPs), 2,4,5- and 2,4,6-trichlorophenol (TCPs) and pentachlorophenol (PCP), as well as those of 1-hydroxypyrene (1-HP) were also measured. Beryllium (Be), manganese (Mn), mercury (Hg) and lead (Pb) concentrations were determined in total blood, while the levels of arsenic (As), cadmium (Cd), chromium (Cr), nickel (Ni) and vanadium (V) were measured in urine. RESULTS: The levels of benzene, toluene, ethylbenzene and m-xylene were under their respective detection limits. The concentrations of HCB, PCBs and PCDD/Fs in plasma of the HWI workers, chlorophenols (CLPs) and 1-HP urinary concentrations, as well as those of metals in blood and urine are shown. PCDD/F concentrations in plasma of the new HWI workers ranged between 13.4 and 84.0 pg international toxic equivalents (I-TEQ)/g lipid, with a geometric mean value of 24.4 pg I-TEQ/g lipid. DISCUSSION: PCDD/F levels, as well as the concentrations of PCBs were of the same order of magnitude as those recently obtained for non-occupationally exposed populations of Catalonia (Spain). No significant gender differences were found for the levels of HCB, PCBs and PCDD/Fs. Although the present results showed a notable variability in the levels of CLPs, geometric mean values of these compounds were similar or even lower than those considered as potential reference values. Metal concentrations in blood and urine were also of the same order of magnitude than previously reported metal levels in the same area. CONCLUSION: The present results should be useful in future surveys in which internal exposure of the HWI workers will be determined.

Adult↗
PubMed2001

Detrusor thickness in healthy children assessed by a standardized ultrasound method.

PURPOSE: We determine the ultrasonographic detrusor thickness in healthy infants and children. MATERIALS AND METHODS: We studied 150 healthy children, 79 boys and 71 girls, from newborns to 13 years old. The detrusor of the ventral and dorsal wall was measured with a previously established ultrasonographic technique, and the variation with age and bladder fullness was assessed. RESULTS: Detrusor thickness varied significantly with the degree of bladder fullness (thinner at higher volumes). It also increased slightly with age. At bladder fullness of 10% or greater of expected bladder capacity (EBC), calculated using the formula EBC (ml.) = age (years) x 30 + 30, a practical upper limit for the detrusor thickness of the ventral wall at all ages was 2.0 mm. at a bladder fullness up to 50% of expected bladder capacity or 1.5 mm. at a higher degree of fullness. The detrusor of the ventral wall was slightly thicker in boys than in girls and also somewhat thicker than the detrusor of the dorsal wall. CONCLUSIONS: The results indicate that ultrasonographic measurements of detrusor thickness in children may serve as a diagnostic tool and its usefulness in clinical as well as research work should be explored.

Adolescent↗
PubMed2001

PCDD/F concentrations in soil and vegetation in the vicinity of a municipal waste incinerator after a pronounced decrease in the emissions of PCDD/Fs from the facility.

Emission of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) by municipal solid waste incinerators (MSWI) is an issue of great concern. In 1997, an adaptation to the EU legislation on pollutant emissions from the stack was carried out in an MSWI from Tarragona (Catalonia, Spain). As a result, PCDD/F emissions were significantly reduced. The aim of this study was to determine the current levels of PCDD/Fs in soil and vegetation samples collected near the facility and to compare these levels with those obtained in previous surveys (1996 and 1997). In the period 1997-1999, PCDD/F concentrations in vegetation samples were significantly decreased (60%). By contrast, the levels of PCDD/Fs in soil samples increased slightly (14%, P > 0.05) during the same period. An exhaustive analysis of the present data indicates that other emission sources of PCDD/Fs have also a notable environmental impact on the area under direct influence of the MSWI.

Air Pollution↗
PubMed2001

Recognition of chronic myelogenous leukaemia cells by autologous T lymphocytes primed in vitro against the patient's dendritic cells.

Defects in immune responses are common in patients with chronic myelogenous leukaemia (CML). However, using dendritic cells (DCs) to promote T-cell immunity in vitro may nonetheless elicit potent specific anti-tumour responses for use in immunotherapy. Here, we show that DCs generated from CML patients had a typical dendritic phenotype and were able to stimulate autologous T cells. Three primed T-cell lines were studied in more detail in one patient. They were stimulated by autologous CML cells, but not by normal non-leukaemic cells from the patient's HLA-identical sibling. This was blocked by HLA-DR-specific, but not HLA-DQ- or HLA-DP-specific antibodies. CML-stimulated cytokine secretion, including interferon-gamma and granulocyte macrophage-colony stimulating factor, suggested a Th1-type phenotype for these sensitized anti-leukaemic T cells. This study therefore shows that cells with a functional dendritic phenotype can be generated from the blood of CML patients and are potent inducers of T-cell responses to tumour cells. This approach allows sensitization of patients' T cells by their own particular tumour without the need to identify the exact leukaemia antigens involved, and may find application in immunotherapy of CML.

Cell Division↗
PubMed2001

Role of the Bacillus subtilis fatty acid desaturase in membrane adaptation during cold shock.

In our attempt to understand the cold shock response of Bacillus subtilis, we report on the role of the B. subtilis fatty acid desaturase (FA-D) Des during membrane adaptation to low temperatures and demonstrate its importance during cold shock. A des null mutant was constructed and analysed in comparison with its parental strain. Growth studies and large-scale comparative fatty acid (FA) analysis revealed a severe cold-sensitive phenotype of the des deletion mutant during the absence of isoleucine and showed that four unsaturated fatty acid (UFA) species differing in length, branching pattern and position of the double bond are synthesized in B. subtilis JH642 but not in the des null mutant. Apart from the lack of UFA synthesis, the FA-D deletion strain showed a dramatically altered saturated fatty acid (SFA) profile at the onset of the stationary growth phase in the presence of exogenous isoleucine sources. Expression of des integrated in trans at the amyE locus of the des deletion strain not only cured the cold-sensitive phenotype observed for the des mutant but allowed much better growth than in strain JH642 after a shift from 37 degrees C to 15 degrees C. These results show that, during cold shock adaptation, des expression can completely replace the isoleucine-dependent, long-term, FA branching adaptation mechanism. We conclude that the crucial aspect in cold adaptation of the cytoplasmic membrane is not its specific molecular composition but rather its physical status in terms of its fluidity.

Adaptation, Physiological↗
PubMed2001

[Variability of coping strategies in coronary artery bypass surgery patients].

The state-versus-trait discussion in coping research would become more meaningful if attention is paid to the variability both of coping patterns in individual patients as well as stability of single coping strategies over time. 35 patients undergoing coronary artery bypass surgery were interviewed three times about their coping and stress experience: after cardiac catheterization, on the day before surgery, and six days after surgery. Anxiety and depression were measured. A coping attitude of "positive passivity" was present at all three points of time. While the group means were stable, vast interindividual differences occurred. Also, there was a high degree of scatter in the stability of single coping items; emotion related coping modes were more stable than cognition and action related ones. The variability of the patients' coping patterns correlated positively with the amount of stress experienced and with preoperative depression. Different coping strategies are linked to a different degree with personality traits, emotional coping modes revealing the closest connection. The variability of individual coping efforts might be linked to a personality disposition characterized by a vulnerability for stress and depressive reactions.

Adaptation, Psychological↗
PubMed2001

[In vitro methods for phototoxicity and photocarcinogenicity testing of drugs].

Phototoxicity is an acknowledged property of some UV and/or visible light absorbing substances some of which are used as pharmaceuticals or in cosmetic preparations. In recent years attention has been called upon the fact that toxic intermediates that are generated upon photoactivation of a substance can also lead to DNA damage. Such damage may lead to mutated/initiated skin cells which in turn can contribute to an elevated skin cancer risk. The method of choice to test for photo-related skin carcinogenesis is a 1-year study in genetically hairless mice in which the formation of skin papilloma and their latency time are assessed. Here, in vitro test approaches to test for photogenotoxicity can be used in a tiered assessment approach asking the use of in vitro genotoxicity tests for prediction of rodent/human carcinogenicity. In the past few years some effort has been put into the evaluation for such systems, in particular standard test protocols have been generated for the in vitro photo-micronucleus test and the in vitro photo-comet assay with Chinese hamster V79 cells. The data that have been produced so far show promising results regarding the implementation of these systems in a tiered approach for photocarcinogenicity assessment of UV- and/or visible light absorbing substances but the systems will have to be validated in further collaborative studies.

Animal Testing Alternatives↗
PubMed2000

Strategies and testing methods for identifying mutagenic risks.

The evolution of testing strategies and methods for identification of mutagenic agents is discussed, beginning with the concern over potential health and population effects of chemical mutagens in the late 1940s that led to the development of regulatory guidelines for mutagenicity testing in the 1970s and 1980s. Efforts to achieve international harmonization of mutagenicity testing guidelines are summarized, and current issues and needs in the field are discussed, including the need for quantitative methods of mutagenic risk assessment, dose-response thresholds, indirect mechanisms of mutagenicity, and the predictivity of mutagenicity assays for carcinogenicity in vivo. Speculation is offered about the future of mutagenicity testing, including possible near-term changes in standard test batteries and the longer-term roles of expression profiling of damage-response genes, in vivo mutagenicity testing methods, and models that better account for differences in metabolism between humans and laboratory model systems.

Carcinogenicity Tests↗
PubMed2000

Evaluating the environmental impact of an old municipal waste incinerator: PCDD/F levels in soil and vegetation samples.

In order to determine the temporal variation in the levels of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) in the vicinity of an old municipal solid waste incinerator (MSWI) (S. Adrià del Besòs, Barcelona, Spain), 24 soil and vegetation samples were collected at the same sampling points in which samples had been taken 1 year before. Each sample was analyzed for PCDDs and PCDFs by high-resolution gas chromatography/high-resolution mass spectrometry. While in the previous study PCDD/F concentrations in soil ranged from 1.22 to 34. 28 ng I-TEQ/kg (median and mean values: 9.06 and 12.24 ng I-TEQ/kg), in the present study, PCDD/F levels ranged from 1.33 to 54.23 ng I-TEQ/kg (median and mean values: 11.85 and 14.41 ng I-TEQ/kg). On the other hand, in the previous study, PCDD/F levels in vegetation ranged from 0.33 to 1.98 ng I-TEQ/kg (median and mean values: 0.58 and 0.70 ng I-TEQ/kg), whereas in the present study, PCDD/F levels ranged from 0.32 to 2.52 ng I-TEQ/kg (median and mean values: 0.82 and 0.97 ng I-TEQ/kg). During the last 12 months, PCDD/F levels increased in 16 of the 24 soil samples and in 17 of the 24 vegetation samples analyzed. However, no significant differences in the median I-TEQ concentrations of both studies were found either in soil or vegetation samples.

Benzofurans↗
PubMed2000

Human biological relevance and the use of threshold-arguments in regulatory genotoxicity assessment: experience with pharmaceuticals.

Issues of biological relevance and thresholds for genotoxicity are discussed here based upon the background of experience with the submissions for the approval of new pharmaceuticals to the German regulatory authority over the period between 1990 and 1997. This experience shows that out of the genotoxicity test systems which are required according to existing guidelines in the European Union (EU), the in vitro tests for chromosomal aberrations (CA) and the mouse lymphoma tk assays (MLA) yield a rate of positives that is about four-fold higher than that of other genotoxicity tests. A detailed analysis of chemical and pharmacological classes of compounds and their effects in these systems reveals that in addition to direct DNA reactivity several mechanisms of indirect genotoxicity such as nucleoside analogue incorporation into DNA, interaction with microtubule assembly, topoisomerase inhibition and high levels of cytotoxicity are relevant. New pharmaceuticals, for which the latter mechanisms apply, often display threshold-like characteristics in their genotoxic effects in vitro or even in vivo in experimental animals. This casts doubt upon the relevance of positive in vitro test results for such compounds. However, the discussion of examples shows that it may not be easy to demonstrate the exact thresholded mechanism of genotoxicity in a given case. In particular, the demonstration of a coincidence of genotoxicity and high levels of cytotoxicity, which seems to be a major factor for biologically non-relevant in vitro positive new pharmaceuticals, usually requires quite extensive testing. Hence, for new pharmaceuticals it is practice to provide in addition to in vitro results that may be thresholded a wealth of information from in vivo studies on genotoxicity, carcinogenicity, metabolism, pharmacokinetics, etc. the results of which help in assessing the biological relevance of in vitro positives. The regulatory acknowledgement of biologically non-relevant, thresholded mechanisms of (in vitro) genotoxicity in addition to those that are considered relevant for human risk ensures a better understanding of test results and is needed for the credibility of genotoxicity testing practice in general.

Animals↗
PubMed2000

Interpretation of the biological relevance of genotoxicity test results: the importance of thresholds.

Despite recent improvements in genotoxicity protocols, we have observed an increase in the occurrence of positive results, particularly in chromosomal aberration tests in vitro, yet very few of these are accompanied by positive responses in vivo. Thus, the positive results may not be biologically relevant either for rodents or humans in vivo, but how should we determine "biological relevance"? Chemicals that produce thresholded dose-responses may well not pose a genotoxic risk at low (relevant to human) exposures, but thresholds should not just be "seen"; there must be an explanation and understanding of the underlying mechanism. In addition to extremes of pH, ionic strength and osmolality, as have been identified previously, such mechanisms include indirect genotoxicity resulting from interaction with non-DNA targets, chemicals/metabolites which are inherently genotoxic but which, at low concentrations, are effectively conjugated and unable to form adducts, and production of specific metabolites under in vitro conditions that are not formed in rodents or humans in vivo. If such thresholded mechanisms can be identified at exposures which are well in excess of expected human exposure, then there may be a strong argument that the positive results are not biologically relevant.

Animals↗
PubMed2000

Considerations on photochemical genotoxicity: report of the International Workshop on Genotoxicity Test Procedures Working Group.

Recent toxicological observations have caused concern regarding the need to test, for example, pharmaceuticals and cosmetic products for photochemical genotoxicity. The objective of this report is to give assistance on how to adapt existing test methods to investigate the potential of light-absorbing compounds to induce genotoxic effects on photoactivation. In general, the Organization for Economic Co-Operation & Economic Development (OECD) draft guideline on in vitro phototoxicity testing served as a basis for consideration. Concomitant exposure of the cells to the test compound and solar simulated light was considered appropriate as the initial, basic test condition. Optimization of the exposure scheme, e.g., a change of the irradiation spectrum, might be indicated depending on the initial test results. Selection of test compound concentrations should be based on results obtained with the dark version of the respective test system but might have to be modified if phototoxic effects are observed. Selection of the irradiation dose has to be performed individually for each test system based on dose-effect studies. The irradiation should induce per se a small, reproducible toxic or genotoxic effect. The report includes a specification of necessary controls, discusses factors that might have an impact on the irradiation characteristics, and gives a rationale for the omission of an external metabolic activation system. It also addresses the question that physicochemical and pharmacokinetic properties might trigger the need to test a chemical for photochemical genotoxicity. Relevant experimental observations are presented to back up the recommendations. The working group did not reach a consensus as to whether a single, adequately perfomed in vitro test for clastogenicity would be sufficient to exclude a photogenotoxic liability or whether a test battery including a gene mutation assay would be needed for product safety testing regarding photochemical genotoxicity.

Animals↗
PubMed2000

Atmospheric deposition of PCDD/Fs near an old municipal solid waste incinerator: levels in soil and vegetation.

The levels of polychlorinated dibenzo-p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDF) were determined in soil and vegetation samples taken from 24 sites in the vicinity of an old municipal solid waste incinerator (San Adrià del Besòs, Barcelona, Spain). Duplicate samples were collected within a radius of 3 km from the stack. PCDD/F concentrations in soils ranged from 1.22 to 34.28 ng I-TEQ/kg (d.m.) with median and mean values of 9.06 and 12.24 ng I-TEQ/kg, respectively. In turn, the levels of PCDD/Fs in vegetation samples ranged from 0.33 to 1.98 ng I-TEQ/kg (d.m.), with median and mean values of 0.58 and 0.70 ng I-TEQ/kg, respectively. Although the present PCDD/F concentrations in soil samples were higher than those recently found in soils taken near other incinerators from Catalonia, they are of the same order of magnitude than the levels of these pollutants found in incinerators from other countries. By contrast, the concentrations of PCDD/Fs in herbage samples were comparable to those found in recent surveys carried out in Catalonia.

Air Pollutants↗