No Concentration Decrease of House Dust Mite Allergens With Rising Altitude in Alpine Regions.

PURPOSE: Several studies over the past 4 decades have indicated a significant reduction in house dust mite (HDM) and HDM allergen concentration in areas higher than 1,500 m above sea level. These have served as basis of allergen avoidance therapies for HDM allergy and asthma. However, modern construction techniques used in the insulation, heating, and glazing of buildings as well as global warming have changed the environmental parameters for HDM living conditions. The present study revisits the paradigm of decreasing HDM allergen concentrations with increasing altitude in the alpine region of Germany and Austria. METHODS: A total of 122 dust samples from different abodes (hotels, privates and mountain huts) at different altitudes (400-2,600 m) were taken, and concentrations of HDM allergens were analyzed. Humidity and temperature conditions, and numerous indoor environmental parameters such as fine dust, type of flooring, age of building, and frequency of cleaning were determined. RESULTS: HDM allergen concentrations did not significantly change with increasing altitude or relative humidity. At the level of indoor parameters, correlations could be found for different flooring types and the concentration of HDM allergens. CONCLUSIONS: In contrast to the widespread view of the relationship between altitude and HDM allergen concentrations, clinically relevant concentrations of HDM allergens could be detected in high-lying alpine regions in Austria and Germany. These results indicate that improvement in conditions of asthmatic patients sensitized against HDMs during a stay at high altitude can no longer be ascribed to decreased levels of HDM allergens, instead, other mechanisms may trigger the beneficial effect.

Effects of ionized waterfall aerosol on pediatric allergic asthma.

OBJECTIVE: Ionized water aerosols have been suggested to exert beneficial health effects on pediatric allergic asthma. Their effect was evaluated in a randomized controlled clinical trial as part of a summer asthma camp. METHODS: Asthmatic allergic children (n = 54) spent 3 weeks in an alpine asthma camp; half of the group was exposed to water aerosol of an alpine waterfall for 1 hour per day, whereas the other half spent the same time at a "control site". Immunological analysis, lung function testing, and fractional exhaled nitric oxide (FeNO) testing were performed during the stay, and sustaining effects were evaluated 2 months later. Symptom score testing was done over a period of 140 days. RESULTS: The water aerosol group showed a significant improvement in all lung function parameters, whereas only the peak expiratory flow improved in the control group. All patients showed a significant improvement in symptom score and a significant decrease in FeNO after the camp. Only the water aerosol group exhibited a long-lasting effect on asthma symptoms, lung function, and inflammation in the follow-up examination. Induction of interleukin (IL)-10 and regulatory T (Treg) cells was measured in both groups, with a pronounced increase in the water aerosol group. IL-13 was significantly decreased in both groups, whereas IL-5 and eosinophil cationic protein were decreased only in the water aerosol group. CONCLUSIONS: Our findings confirm the induction of Treg cells and reduction in inflammation by climate therapy. They indicate a synergistic effect of water aerosols resulting in a long-lasting beneficial effect on asthma symptoms, lung function, and airway inflammation.

Low dose hypericin-PDT induces complete tumor regression in BALB/c mice bearing CT26 colon carcinoma.

BACKGROUND: Successful tumor eradication with photodynamic therapy (PDT) in vivo depends on the optimal combination of treatment parameters. (Low-dose) PDT may additionally induce antitumoral immune responses. Since the naturally occurring hypericin (Hyp) is a promising photosensitizer for PDT, the aim of the study was to investigate phototoxic and immunologic effects of a low-dose Hyp-PDT on murine tumors in contrast to commonly used Hyp-PDT conditions. METHODS: BALB/c mice bearing CT26 colon carcinoma received hypericin intravenously and were irradiated with red light 0.5-4h later. Tumor development was recorded. Mice were then re-challenged 60 days after the first tumor cell inoculation to investigate an antitumoral immune response. RESULTS: Different results of tumor/host responses were obtained, ranging from mice exitus over delayed tumor growth to complete tumor regression according to different treatment protocols. PDT with common doses and a 4h drug-light-interval resulted in a four times delayed tumor growth compared to the control groups. PDT with relatively low doses and a drug-light-interval of 0.5h led to 100% tumor eradication. Re-challenge of these mice with CT26 mouse colon carcinoma cells prevented new tumor growth. CONCLUSIONS: Not only drug concentrations and light doses seem to determine the efficiency of tumor eradication, but also the localization of hypericin at the time of irradiation. Targets in our low-dose PDT protocol are exclusively the vessels. The advantage of this low-dose PDT beside less drug and light exposure of the animals is reduced skin damage, faster healing of the lesions and induction of an antitumoral immune response.

Time-resolved gene expression profiling of human squamous cell carcinoma cells during the apoptosis process induced by photodynamic treatment with hypericin.

Hypericin is used as a powerful naturally occurring photosensitizer in photodynamic therapy (PDT). Activated by visible light, it kills tumour cells and tissues via generation of reactive oxygen species (ROS). Depending on the protocol, apoptotic cell death can be achieved very effectively by hypericin-PDT. To analyze the fundamental molecular mechanisms leading to apoptosis induced by photodamage especially with regard to human skin cancer cells, we studied the alteration of the gene expression pattern in the human squamous cell carcinoma cell line A-431 at 1.5, 3, 5 and 8 h after hypericin-PDT by cDNA-macroarray technique. Radioactively labelled samples were hybridized onto macroarray filters containing PCR products of 9738 ESTs of the Incyte Human UniGEM Microarray clone set. In total, 168 genes were found to be differentially upregulated and 45 down-regulated. Verification of expression changes of 45 genes of interest was performed by quantitative real-time PCR. Due to the observed significant expression changes the following can be concluded: lipoprotein receptor-mediated endocytosis could play a role in the uptake of lipophilic hypericin. Extracellular signal transduction to the cell is reduced, cell detachment facilitated, changes of the morphology, cytoskeleton and formation of apoptotic bodies occur. The promotion of p38MAPK, ERK, JNK and Ras signalling pathways supports survival and/or apoptosis. Switches between life and death could be the strongly upregulated transcription factors c-jun and FOSB as well as the MAPK-phosphatase 1 DUSP-1, possibly activated via H3 histone modifications. ROS activate ER-stress pathways or adaptive response, and provoke damage protection against ROS, partly in a cell-type specific way.

Gene expression profiling of the human carcinoma cell line A-431 after 5-aminolevulinic acid-based photodynamic treatment.

The photosensitizer protoporphyrin IX, endogenously accumulated from the precursor aminolevulinic acid (ALA) as well as other less photodynamically effective intermediates, is a successful agent in photodynamic therapy. Despite encouraging clinical results, the basic mechanisms leading to cell death are not fully understood. To elucidate these fundamentals, the alteration of the gene expression pattern in the squamous cell carcinoma cell line A-431 was studied at different time-points after photodynamic treatment with ALA by cDNA-array technique. Cells were incubated for 16 h with 100 microg/ml ALA and irradiated with a fluence of 3.5 J/cm(2) resulting in 50% survival until 8 h post treatment. RNA was isolated at 1.5, 3, 5 and 8 h post treatment and from 3 controls (untreated, light only and dark), radioactively labeled by reverse transcription with 33P-dCTP and hybridized onto macroarray PCR filters containing PCR products of 2135 genes, which were selected for relevance in carcinogenesis, stress response and signal transduction. Verification of observed expression changes was carried out by real-time RT-PCR. We found a strong induction of expression of the immediate early genes c-jun and c-fos as well as decreased expression of genes involved in proliferation such as myc, genes involved in apoptosis such as Fas associated via death domain (FADD) and the fibronectin gene for cell adhesion. An apoptosis induction rate of not more than 30% as proved by apoptosis detection assays and caused by PpIX localization in the membrane was reflected by the expression profile.

Gene expression pattern following photodynamic treatment of the carcinoma cell line A-431 analysed by cDNA arrays.

The photosensitizer protoporphyrin IX, endogenously accumulated from the precursor aminolevulinic acid, is a successful agent in photodynamic tumor therapy. In spite of encouraging clinical results, the basic mechanisms leading to cell death are not fully understood. We therefore set out to analyse the alteration of the gene expression pattern in the squamous cell carcinoma cell line A-431 after photodynamic treatment with endogenous protoporphyrin IX. Radioactively labelled cDNAs from untreated and treated cells were hybridized onto UniGene cDNA array filters containing lysed bacterial colonies with inserts representing approximately 32000 different human transcripts. Differentially expressed genes were identified and verified on sub-arrays containing only the candidate genes. We found increased expression of heat shock protein 70 and of the immediate early genes p55-c-fos and c-jun, may be due to oxidative stress and increased levels of intracellular calcium after photoactivation of protoporphyrin IX. Increased expression of heme oxygenase-1 following dark incubation was not further increased by irradiation and therefore probably caused by the need for heme degradation. Presumably heat shock protein 70 and heme oxygenase-1 serve for cell protection. Though similar results have been found for photodynamic treatment with external porphyrin-based photosensitizers, this is the first report about induction of the genes described above by (photoactivated) endogenous protoporphyrin IX.