The release and transport of fungal spores from water-damaged building materials is a key factor for understanding the exposure to particles of fungal origin as a possible cause of adverse health effects associated to growth of fungi indoors. In this study, the release of spores from nine species of typical indoor fungi has been measured under controlled conditions. The fungi were cultivated for a period of 4-6 weeks on sterilized wet wallpapered gypsum boards at a relative humidity (RH) of approximately 97%. A specially designed small chamber (P-FLEC) was placed on the gypsum board. The release of fungal spores was induced by well-defined jets of air impacting from rotating nozzles. The spores and other particles released from the surface were transported by the air flowing from the chamber through a top outlet to a particle counter and sizer. For two of the fungi (Penicillium chrysogenum and Trichoderma harzianum), the number of spores produced on the gypsum board and subsequently released was quantified. Also the relationship between air velocities from 0.3 to 3 m/s over the surface and spore release has been measured. The method was found to give very reproducible results for each fungal isolate, whereas the spore release is very different for different fungi under identical conditions. Also, the relationship between air velocity and spore release depends on the fungus. For some fungi a significant number of particles smaller than the spore size were released. The method applied in the study may also be useful for field studies and for generation of spores for exposure studies.

The serine-protease hepsin is one of the most prominently overexpressed genes in human prostate carcinoma. Forced expression of the enzyme in mice prostates is associated with matrix degradation, invasive growth, and prostate cancer progression. Conversely, hepsin overexpression in metastatic prostate cancer cell lines was reported to induce cell cycle arrest and reduction of invasive growth in vitro. We used a system for doxycycline (dox)-inducible target gene expression in metastasis-derived PC3 cells to analyze the effects of hepsin in a quantitative manner. Loss of viability and adhesion correlated with hepsin expression levels during anchorage-dependent but not anchorage-independent growth. Full expression of hepsin led to cell death and detachment and was specifically associated with reduced phosphorylation of AKT at Ser(473), which was restored by growth on matrix derived from RWPE1 normal prostatic epithelial cells. In the chorioallantoic membrane xenograft model, hepsin overexpression in PC3 cells reduced the viability of tumors but did not suppress invasive growth. The data presented here provide evidence that elevated levels of hepsin interfere with cell adhesion and viability in the background of prostate cancer as well as other tissue types, the details of which depend on the microenvironment provided. Our findings suggest that overexpression of the enzyme in prostate carcinogenesis must be spatially and temporally restricted for the efficient development of tumors and metastases.

A conceptual approach including measurements of materials at rest (step 1), measurements using a large rotating drum (step 2) or a Particle-FLEC (step 2) and measurements at a workplace (step 4) has been used to characterize the release of microbial components (bacteria, fungi, actinomycetes, endotoxin or enzymes) and particles from straw, wood chips or fungal cultures of different ages on gypsum boards. Repeated agitation or handling periods were included in step 2 and step 4. There was a low similarity between the amount of microbial components measured in step 1 and the aerosolized amount (step 2) from gypsum boards, wood chips and straw. Ratios between some microbial components measured at the workplace (step 4) and measured in step 2, showed similarities. Less than 1.3% of the total amount of microorganisms and endotoxin becomes airborne during 5 min of agitation of straw or wood chips. Most microbial components were released at higher rates during the first agitation period than during the following periods. However, differences were seen between different microbial components, and endotoxin from straw was released at the same rate in two successive agitation periods. Fungal particles smaller than spores were released from fungal colonized gypsum boards at amounts that were up to 30 times higher in the first agitation period compared with that in the following period, while fungal spores were released at amounts that were five times as high in the first period compared with that in the following period. In addition to differences between microbial components, the release patterns of microbial components were different for wood chips and straw. The time for maximum particle release to half particle release was longer for straw than for wood chips. The observation that some components, e.g. endotoxin, are released at the same rate in two successive handling steps, and that others (e.g. fungi) are mainly released initially, shows that the exposure period to different components from the same material differs in duration. The observed differences in the release patterns of different components and the differences between materials are important when preventive steps are to be taken, and it stresses the importance of applying a relevant sampling time and period in exposure assessments.

A rapid, precise, and viability‐retaining method for cytoplasmic molecule delivery is highly desired for cell engineering. Routine methods suffer from low throughput, lack of selectivity, requirement of helper compounds, predominant endosomal delivery, and/or are restricted to specific molecule classes. Photonic cell manipulation bears the potential to overcome these drawbacks. Here we investigated mammalian cell manipulation by single sub‐nanosecond laser pulses. Axial beam waist positioning close to a cell monolayer induced culture vessel damage and zones of cell ablation. Cells at margins of ablation zones exhibited uptake of membrane‐impermeant fluorophores and GFP expression plasmids. Increasing Rayleigh‐length and beam waist diameter reduced the sensitivity to axial defocusing and resulted in robust molecule transfer. Serial application of single pulses focused over a moving cell monolayer yielded quantitative molecule transfer to cells at rates up to 40%. Our results could be basic to spatially and temporally controlled single laser pulse‐mediated marker‐free high throughput cell manipulation.