Development and performance assessment of a qualitative SYBR® green real-time PCR assay for the detection of Aspergillus versicolor in indoor air.

Currently, contamination of indoor environment by fungi and molds is considered as a public health problem. The monitoring of indoor airborne fungal contamination is a common tool to help understanding the link between fungi in houses and respiratory problems. Classical analytical monitoring methods, based on cultivation and microscopic identification, depend on the growth of the fungi. Consequently, they are biased by difficulties to grow some species on certain culture media and under certain conditions or by noncultivable or dead fungi that can consequently not be identified. However, they could have an impact on human health as they might be allergenic. Since molecular methods do not require a culture step, they seem an excellent alternative for the monitoring of indoor fungal contaminations. As a case study, we developed a SYBR® green real-time PCR-based assay for the specific detection and identification of Aspergillus versicolor, which is frequently observed in indoor environment and known to be allergenic. The developed primers amplify a short region of the internal transcribed spacer 1 from the 18S ribosomal DNA complex. Subsequently, the performance of this quantitative polymerase chain reaction (qPCR) method was assessed using specific criteria, including an evaluation of the selectivity, PCR efficiency, dynamic range, and repeatability. The limit of detection was determined to be 1 or 2 copies of genomic DNA of A. versicolor. In order to demonstrate that this SYBR® green qPCR assay is a valuable alternative for monitoring indoor fungal contamination with A. versicolor, environmental samples collected in contaminated houses were analyzed and the results were compared to the ones obtained with the traditional methods.

The role of indoor pollution in the development and maintenance of chronic airway inflammation in children.

Air pollution is one of the great problems of this century and it plays an important role in the increasing prevalence of chronic inflammatory problems in the upper airway in children. Since their lungs and immune system are not fully developed when exposure begins, newborns and children appear to be more sensitive to the effects of both outdoor and indoor air pollution. Furthermore, children spend most of their time indoors and are exposed more often to pollutants in indoor air. The link between health problems, chemical products and allergens (the latter mainly from cats and mites) has been extensively studied. Other important indoor contaminants are fungi, which are often present in damp buildings and can cause severe respiratory disease by producing spores, allergens, volatile irritant compounds and toxins. A proper identification of mould contamination of this kind is vital for correct diagnosis, treatment and the prevention of health problems, and improvements have been observed after the removal or cleaning of the contaminated materials and improvements to the ventilation of buildings. While a possible association between respiratory symptoms, such as rhinitis, and the presence of fungi in the indoor environment has been documented by several authors, other studies have observed no significant relationship. The development of standardised sampling, detection and diagnostic tests will be essential to understand the proper role of fungi in the indoor atmosphere and their impact on public health.

Inhibition of MAO by fractions and constituents of hypericum extract.

The inhibition of monoamine oxidase (MAO) by six fractions from hypericum extract and three characteristic constituents (as pure substances) were analyzed in vitro and ex vivo to study the antidepressive mechanism of action. Rat brain homogenates were used as the in vitro model, while the ex vivo analysis was performed after intraperitoneal application of the test substances to albino rats. Massive inhibition of MAO-A could be shown with the total extract and all fractions only at the concentration of 10(-3) mol/L. At 10(-4) mol/L, one fraction rich in flavonoides showed an inhibition of 39%, and all other fractions demonstrated less than 25% inhibition. Using pure hypericin as well as in all ex vivo experiments, no relevant inhibiting effects could be shown. From the results it can be concluded that the clinically proven antidepressive effect of hypericum extract cannot be explained in terms of MAO inhibition.

Pharmaceutical quality of hypericum extracts.

Hypericum extracts contain at least ten constituents or groups of components that may contribute to the pharmacological effects. It is not yet possible to correlate the antidepressive mode of action with specific constituents; therefore, the pharmaceutical quality of the extracts was characterized on the basis of typical leading substances and especially the hypericins. For the analysis and improvement of the production procedure, the content of hypericin and pseudohypericin was measured experimentally. The drug material was extracted with different solvents and the yield was analyzed for each kind of solvent, its concentration and extraction temperature. Optimal yields were obtained with 80% methanol at temperatures of 80 degrees C.

Analytical and pharmacological studies on Mahonia aquifolium.

Alcoholic extracts of the antipsoriatic Mahonia aquifolium drug which were analysed for their qualitative and quantitative alkaloid content, showed an in vitro inhibiting effect on cyclo-oxygenase (CO) of sheep seminal vesicles and 5-lipoxygenase (5-LO) of porcine leucocytes. The same extracts exhibited also immunostimulating activities in two phagocytosis assays. In contrast the major alkaloids of Mahonia aquifolium extracts berberine, palmatine, magnoflorine and jatrorrhizine were found to be inactive at all in both antiinflammatory assays, suggesting a different mechanism of action for the alkaloids or further not yet detected compounds in the extract.

[DC- and HPLC-Analysis of Eleutherococcus.]. / Die DC- and HPLC-Analyse der Eleutherococcus Droge.

A rapid and reproducible TLC- and HPLC-analysis of root-extracts and preparations of ELEUTHEROCOCCUS SENTICOSUS on the basis of the characteristic lignans and other phenylpropan derivatives has been developed.