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.
The effects of low-temperature plasma treatment on microorganisms typically related to skin diseases are studied qualitatively by the inhibition of growth and viability assays to evaluate the potential for classifying as a prospective antiseptic agent. A variety of microorganisms enveloping gram- negative and gram-positive bacteria as well as one genus of yeast and fungus each were exposed to plasma in vitro. In a comparative approach, two power supplies, both of which produce high voltage pulses yet at different temporal characteristics, are applied for the growth study. While operation with both devices led to growth inhibition of all microbes, the results indicate a superior antimicrobial efficacy for high voltage pulse lengths in the nanosecond scale. Fluorescence assays reveal the efficacy of nanosecond-pulse driven plasma in reducing germ viability. Furthermore, the technical background for patents related to low-temperature plasma technology in the field of plasma medicine is discussed.
A new, gentle enzymatic method was developed for a controlled removal of casein layers from medieval wall paintings. These casein layers were applied over the last 60 years on wall paintings in order to decrease substantial damage due to a peeling off of the frescoes from the roughcast surface due to environmental effects. However, due to the aging of the casein layers (at 40±50 years), a more drastic peeling occurred and the danger of total destruction of the wall paintings is severe. Thus, screening was performed to ®nd the most suitable enzyme for casein digestion. Alcalase 2.5 DX L was the most appropriate enzyme for an effective proteolysis reaction. The enzyme was immobilized on functionalized cellulose membrane. A membrane pad system with immobilized enzymes was developed which could be pressed on the casein layers on the wall painting. A controlled removal of the casein layers by proteolytic digestion was observed and it was possible to continuously wash off the hydrolyzed casein fragments from the wall painting surface by an aqueous carbonate buffer¯owing through the membrane pad. The removal and the digestion was monitored by reverse HPLC. Additionally, an on-line monitoring system was set up in order to continuously follow the casein layer removal and the digestion procedure directly on the wall painting. This technique is based on noninvasive 2D-¯uorescence monitoring. Optical ®ber systems were used to continuously monitor the¯uorescence intensity of casein-bound tryptophan. The off-line data were veri®ed with the on-line 2D-¯uorescence data. Based on the scienti®c result an appropriate technique for the controlled enzymatic removal of damaging casein layers on the surface of medieval wall paintings using immobilized enzyme is now available. It is now applied to remove such casein layers from medieval wall paintings in the Allerheiligen-Kapelle Cloister,
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