Cathelicidin related antimicrobial peptide, laminin, Toll-like receptors and chemokines levels in experimental hypersensitivity pneumonitis in mice.

INTRODUCTION: Hypersensitivity pneumonitis (HP) is an interstitial lung disease caused by unresolved inflammation and tissue repair pathologies triggered by repeated organic dust exposure. The aim of the study was to investigate changes in levels of the cathelicidin related antimicrobial peptide (CRAMP), laminin (LAM-A1), selected Toll-like receptors (TLR) and chemokines in experimental HP in mice. MATERIALS AND METHODS: Three and 18-month-old female C57BL/6J mice underwent inhalations of the saline extract of Pantoea agglomerans cells, Gram-negative bacterium common in organic dust and known for its pathogenic impact. The inhalations were repeated daily (28 days). ELISA was used for measuring in lung tissue homogenates concentration of CRAMP, LAM-A1, TLR2, TLR4, TLR8, CXCL9 (chemokine [C-X-C motif] ligand) and CXCL10. RESULTS: Levels of TLR2, TLR4 and CXCL9 were significantly higher in both young and old mice lungs already after 7 days of inhalations, while significant increase of LAM-A1 and CXCL10 was noted after 28 days, compared to untreated samples. TLR8 level was significantly augmented only in young mice. Only CRAMP level significantly declined. Significantly higher TLR8 and CXCL9 concentration in untreated samples were noted in old animals compared to young ones. CONCLUSION: Significant alterations of the examined factors levels indicate their role in HP pathogenesis.

Cathelicidin LL-37 in bronchoalveolar lavage and epithelial lining fluids from COPD patients and healthy individuals.

Innate immunity is currently under scope of interest concerning its role in the development of chronic obstructive pulmonary disease (COPD). Antimicrobial peptides constitute a potent part of this fast response system. Here, we focus on the role of a specific antimicrobial peptide, the only human cathelicidin, the pleiotropic LL-37 peptide, in the development of COPD under clinical conditions. A cross-sectional study was conducted in groups of 43 patients with COPD (previously classified according to GOLD) and 12 healthy individuals. Bronchoalveolar lavage fluid (BALF) sampling, followed by LL-37 measurements by mass spectrometry combined with previous immunoaffinity purification, was performed. Based on urea levels, concentrations of LL-37 in epithelial lining fluid (ELF) were calculated. Additionally, an antimicrobial assay of growth inhibition of two bacterial species, often involved in COPD development mechanisms, by purchased LL-37 was conducted. Altogether, 55 BALF samples were analyzed. LL-37 levels were significantly higher in BALF from patients in early stages of COPD (GOLD I-II) compared to BALFs from healthy individuals. The same was true for ELF. Cathelicidin’s concentration was significantly lower in both BALF and ELF from patients in advanced COPD (GOLD III-IV). The significantly elevated LL-37 levels both in BALF and ELF in patients with COPD at stage GOLD I-II together with reduced levels in advanced (COPD stage III-IV) further supports the innate immunity involvement in COPD pathology and suggests a profound change in non-specific immunity during the disease progression.

Exposure to airborne microorganisms in fiberboard and chipboard factories.

Microbiological air sampling was performed in one fiberboard factory and two chipboard factories located in south-eastern Poland. It was found that the levels of bacteria, fungi, dust and bacterial endotoxin in the air of examined facilities were high during initial stages of the production cycle (shredding of waste wood, storing of chips) and then sharply decreased during further stages of this cycle (forming and formatting of the boards). In the fiberboard factory, the concentration of airborne microorganisms at the initial stages of production cycle was 71.8-95.2 x 10(3) cfu/m3 and dropped in further stages to the level of 8.4-17.5 x 10(3) cfu/m3. Fungi (mostly Aspergillus fumigatus and Penicillium spp.) were prevailing microorganisms in the air of the fiberboard factory, forming 46.0-87.3% of the total airborne microflora. The concentrations of microorganisms in the air of the chipboard factories were significantly lower compared to the fiberboard factory (p<0.05). During initial stages of production cycle they were within the range of 12.9-101.5 x 10(3) cfu/m3, while during forming and formatting of boards within the range of 5.3-12.4 x 10(3) cfu/m3. On average, the most common microorganisms in the air of the chipboard factories were corynebacteria (mostly Arthrobacter spp. and Corynebacterium spp.) which formed 24.4-64.6% of the total microflora. The values of the respirable fraction of airborne microflora in the fiberboard and chipboard factories varied within a fairly wide range and were between 20.5-91.1%. Altogether, 38 species or genera of bacteria and 16 species or genera of fungi were identified in the air of examined factories, of which respectively 14 and 9 species or genera were reported as having allergenic and/or immunotoxic properties. The concentration of bacterial endotoxin in the air of examined factories was greatest, similarly to the concentration of microorganisms, during the initial stages of the production cycle: 103.1-1974.0 EU/m3 in the fiberboard factory, and 3.2-217.4 EU/m3 in chipboard factories. In conclusion, the workers of fiberboard and chipboard factories may be exposed during the initial stages of the production cycle (shredding of waste wood, storing of chips) to high levels of airborne microorganisms and endotoxin posing respiratory hazard.

Exposure to airborne microorganisms and endotoxin in herb processing plants.

Microbiological air sampling was performed in two herb processing plants located in eastern Poland. Air samples for determination of the levels of bacteria, fungi, dust and endotoxin were collected at 14 sites during cleaning, cutting, grinding, sieving, sorting and packing of 11 kinds of herbs (nettle, caraway, birch, celandine, marjoram, mint, peppermint, sage, St. John's wort, calamus, yarrow), used for production of medications, cosmetics and spices. It was found that processing of herbs was associated with a very high pollution of the air with bacteria, fungi, dust and endotoxin. The numbers of microorganisms (bacteria and fungi) in the air of herb processing plants ranged within 40.6-627.4 x 10(3) cfu/m3 (mean +/- S.D = 231.4 +/- 181.0 x 10(3) cfu/m3). The greatest concentrations were noted at the initial stages of production cycle, during cleaning, cutting and grinding of herbs. The numbers of airborne microorganisms were also significantly (p<0.0001) related to the kind of processed herb, being the greatest at processing marjoram, nettle, yarrow and mint. The values of the respirable fraction of airborne microflora in the examined facilities varied within a fairly wide range and were between 14.7-67.7%. The dominant microorganisms in the air of herb processing plants were mesophilic bacteria, among which endospore-forming bacilli (Bacillus spp.) and actinomycetes of the species Streptomyces albus were most numerous. Among Gram-negative bacteria, the most common was endotoxin-producing species Alcaligenes faecalis. Altogether, 37 species or genera of bacteria and 23 species or genera of fungi were identified in the air of herb processing plants, of these, 11 and 10 species or genera respectively were reported as having allergenic and/or immunotoxic properties. The concentrations of dust and bacterial endotoxin in the air of herb processing plants were large with extremely high levels at some sampling sites. The concentrations of airborne dust ranged within 3.2-946.0 mg/m3 (median 18.1 mg/m3), exceeding at 13 out of 14 sampling sites the Polish OEL value of 4 mg/m3. The concentrations of airborne endotoxin ranged within 0.2-2681.0 microg/m3 (median 16.0 microg/m3), exceeding at all sampling sites the suggested OEL value of 0.1 microg/m3. In conclusion, the workers of herb processing plants could be exposed to large concentrations of airborne microorganisms, dust and endotoxin posing a risk of work-related respiratory disease.

Response of herb processing workers to work-related airborne allergens.

A group of 51 herb processing workers employed in a big herb processing facility located in eastern Poland were examined by the skin and precipitin tests with, respectively, 4 and 17 extracts of microorganisms associated with organic dusts. Out of this number, 32 workers were examined by the skin test with 7 extracts of selected herbs processed in the facility. All the subjects were asked about the occurrence of work-related symptoms. 32 healthy office workers were examined with microbial extracts as a reference group. The herb processing workers showed a high proportion of early skin reactions (after 20 min) to the extract of Gram-negative bacterium Alcaligenes faecalis (41.2%), significantly higher compared to the reference group (p<0.01). At all time intervals (20 min, 8 hrs, 24 hrs), the workers responded with a high frequency to the extract of Bacillus subtilis (respectively 72.5%, 64.7%, and 15.7%), significantly greater compared to the reference group (respectively p<0.001, p<0.001, and p<0.05). No significant differences were found between the groups of herb processing workers and referents in skin response to the extracts of Streptomyces albus and Alternaria alternata and, except for the extract of Pantoea agglomerans, in the frequency of positive precipitin reactions to microbial antigens. In the skin test with herb extracts, the highest response among workers were caused by the extracts of chamomile flowers and nettle leaves which evoked 40-65% of positive skin reactions at all time intervals. 39 out of 51 interviewed herb processing workers (76.5%) reported the occurrence of work-related general, respiratory and skin symptoms. The positive skin reactions occurred more frequently among symptomatic workers which suggests that the specific immunologic response might be implicated in etiopathogenesis of work-related symptoms in examined workers. However, in most cases the differences did not attain a significance level which indicates that there is no direct relationship between a positive immunologic response and the appearance of symptoms caused by occupational exposure to herb dust, and that most probably a considerable part of these symptoms might be also due to non-specific immunologic and/or toxic mechanisms.

Exposure to airborne microorganisms in Polish sawmills.

Microbiological air sampling was performed in four sawmills located in eastern Poland, of which two were processing coniferous wood (pine, fir) and other two deciduous wood (oak, birch). Total concentration of microorganisms (bacteria and fungi) in the air of sawmills processing coniferous wood was on average 20.2 +/- 5.6 x 10(3) cfu/m(3) (mean +/- S.E.) and significantly (p < 0.05) higher compared to those processing deciduous wood where the mean concentration of airborne microorganisms was 9.8 +/- 3.0 x 10(3) cfu/m(3). The greatest concentrations of microorganisms in the sawmills processing coniferous wood were noted at debarking and at first-cut frame sawing of pine logs (42.1 +/- 7.6 x 10(3) cfu/m(3) and 39.8 +/- 7.0 x 10(3) cfu/m(3), respectively). Microflora released into air during debarking consisted mostly of allergenic fungi (mainly Aspergillus fumigatus) and corynebacteria, whereas airborne microflora recovered during first-cut frame sawing constituted mostly of endotoxin-producing Gram- negative bacteria of the genus Rahnella, developing in the sapwood of pine. In the sawmills processing deciduous wood, the largest concentration of microorganisms (30.6 +/- 3.4 x 10(3) cfu/m(3)) was found at sorting of the oak parquet boards and was due to the secondary infection of the boards with moulds Penicillium citrinum during prolonged storing in the open air. Values of the respirable fraction of airborne microflora in the examined sawmills varied within fairly wide limits and were between 22.5 86.6%. Altogether, 34 species or genera of bacteria and 21 species or genera of fungi were identified in the air of sawmills, of which respectively 13 and 9 species or genera were reported as having allergenic and/or immunotoxic properties. The concentrations of airborne bacterial endotoxin which were determined on two sampling sites in the sawmills processing pine and fir, were 0.24 microg/m(3) and 4.00 microg/m(3) respectively, distinctly exceeding the suggested safe level. In conclusion, the workers of Polish sawmills may be exposed on some working stands to airborne microorganisms posing respiratory hazard, of which the greatest risk is represented by allergenic fungi developing on bark of logs or stored wood products and endotoxin-producing Gram-negative bacteria of the genus Rahnella, developing in sapwood of coniferous logs

Response of sawmill workers to work-related airborne allergens.

The aim of this work was to determine the reactivity of sawmill workers to biological allergens associated with wood dust. Allergological examinations by skin and precipitin tests were performed in 43 workers employed in a sawmill processing coniferous wood (pine), in 90 workers employed in two sawmills processing deciduous wood (oak), and in 32 healthy urban dwellers not exposed to organic dusts (referents). The skin test was performed by the intradermal method with the saline extracts of wood dust and of the cultures of three microbial species (Rahnella sp., Brevibacterium linens and Penicillium citrinum) isolated from the air polluted with wood dust. Sawdust from pine was used for testing of the pine processing workers and referents while sawdust from oak was used for testing of the oak processing workers. Skin reactions were recorded after 20 minutes, 8 hours and 24 hours. The agar-gel test for the presence of precipitins in serum was performed with the extract of pine wood dust and extracts of 17 microbial isolates. The workers processing pine showed a very high frequency of positive skin reactions to the extract of wood dust at all time intervals, significantly greater compared to the workers processing oak and referents (p < 0.001). The early skin reactions to the extracts of dust-borne bacteria and fungi were very common among sawmills workers and showed a significant relationship with the degree of exposure. The frequency of reactions to Gram-negative bacterium Rahnella sp. was significantly greater in the pine processing workers than in the oak processing workers and referents (p < 0.001). By contrast, the oak processing workers reacted significantly more frequently to Penicillium citrinum, compared to the pine processing workers and referents (p < 0.01). These results conform to the prior study of airborne microflora in which the dominancy of Gram-negative bacteria was stated in the pine processing sawmill while mould fungi were most common in the oak processing sawmills. The antibody response of sawmill workers to work-related antigens was much weaker compared to skin reactions. As many as 41 sawmill workers reported the occurrence of work-related symptoms. A significant relationship was found between the occurrence of symptoms and frequency of allergic reactions, but only with a limited number of antigens. The obtained results suggest that early allergic reactions to coniferous wood and to microorganisms associated with wood dust are common among sawmill workers, posing a potential risk of work-related disease in this occupational group.

Occupational biohazards in agricultural dusts from India.

Sixteen samples of settled dusts deposited during handling of various granular plant materials (green gram, red gram, amaranth, rice, pearl millet, sorghum, wheat, maize) in small food storing and processing facilities (godowns) were collected in the region of Aurangabad (Southern India). The samples were examined by the dilution plating method for the concentration and species composition of Gram-positive mesophilic bacteria, Gram-negative mesophilic bacteria, thermophilic actinomycetes and fungi. They were also examined by Limulus test for the concentration of bacterial endotoxin. The total concentration of microorganisms (bacteria + fungi) in examined samples varied within a wide range of 1.4 x 10(5) - 8.45 x 10(8) cfu/g (median 8.36 x 10(6) cfu/g). On average, the most common were Gram-positive bacteria (87.84% of all isolates) followed by Gram-negative bacteria (11.12%). Less common were fungi (1.24%) and thermophilic actinomycetes (0.01%). Among isolated bacteria and fungi, there were many species known as causative agents of allergic alveolitis, asthma and organic dust toxic syndrome. The concentration of bacterial endotoxin in the examined samples ranged between 12.5 - 62500 microg/g (median 781.25 microg/g), being particularly large in the samples of dust from maize (6250 microg/g and 62500 microg/g) and pearl millet (6250 microg/g and 12500 microg/g). The results of the present work indicate that the agricultural dusts from India represent a potential hazard for the workers because of high concentrations of allergenic microorganisms and bacterial endotoxin. The particular risk is associated with handing of maize and pearl millet. Further studies on this subject with the use of aerobiological methods are highly desirable

Influence of some immune factors on the IL-6 and soluble IL-2 receptors in haemodialysed patients.

In respect to the immune deficiency state of long-term haemodialysed patients, both cytokines and their receptor disturbances have been taken into consideration. The purpose of our study was to evaluate the effect of uraemic and haemodialysis factors on the interleukin-6 and interleukin-2 soluble receptor levels and the reactivity after influenza vaccination. We have found that IL-6 and IL-2 receptor levels were statistically significantly elevated (98.8 +/- 39 pg/ml and 1557 +/- 544 U/ml, respectively) in serum of haemodialysed patients. The fact that increased immune complexes statistically correlated with soluble IL-2 receptor levels (p < 0.01) was very interesting for us. In order to study the immunological response after vaccination, 10 patients have been investigated after influenza vaccination. Plasma samples were collected before, as well as 1 and 4 weeks after vaccine administration. Antibody titres measured by haemagglutinin inhibition showed decreased antibody levels in haemodialysed patients. We conclude that the interleukin disturbance and the elevated interleukin-2 receptor levels together with the presence of circulating immune complexes can influence in some way the immune response of haemodialysed patients.

[Microflora of the air in sawmills as a potential occupational hazard: concentration and composition of microflora and immunologic reactivity of workers to microbial aeroallergens]. / Mikroflora powietrza tartaków jako potencjalny czynnik zagrozenia zawodowego: stezenie i sklad mikroflory oraz immunologiczna reaktywnosc pracowników na aeroalergeny drobnoustrojowe.

Microbiologial studies of the air and allergological examinations of the workers were performed in two sawmills processing deciduous wood (mainly oak) and in one sawmill processing coniferous wood (mainly pine). The concentration of microorganisms in the air was of the order 10(3)-10(4) cfu/m3. The most common organisms were corynebacteria (Arthrobacter, Corynebacterium, Brevibacterium, Microbacterium), spore-forming bacilli (Bacillus), Gram-negative bacteria (rahnella) and filamentous fungi (Aspergillus, Penicillium). The workers responded to the extract of pine dust with much higher frequency than to the extract of oak dust. The workers processing pine were often sensitized to Rahnella while those processing oak were commonly sensitized to Penicillium. Precipitin reactions were rare and occurred only with the antigen of Rahnella.

[The effect of wood dust on the respiratory system. Medical examination of furniture factory workers]. / Wplyw pylu drzewnego na uklad oddechowy. Badania lekarskie pracowników zakladów meblarskich.

The medical-environmental questionnaire, physical examination and pre-shift and post-shift spirometry have been performed in 48 furniture factory workers. The workers showed the work-related symptoms: cough, shortness of breath, chest pain, headache, general malaise, skin symptoms, eye symptoms, rhinitis. No relationship was found between the spirometry values and the frequency of the symptoms. The exposed workers showed a significant post-shift reduction of the FVC, FEV1, FEV1%VC and PEF (p < 0.001). The higher drops of the spirometric parameters occurred in younger workers. The presented data show that processing of wood may be associated with the work-related respiratory symptoms and diseases in exposed workers.

[Microflora of the in furniture factors as a potential occupational hazard: concentration and composition of microflora and immunologic reactivity of workers to microbial aeroallergens]. / Mikroflora powietrza fabryk mebli jako potencjalny czynnik zagrozenia zawodowego: stezenie i skiad mikroflory oraz immunologiczna reaktywnosc pracowników na aeroalergeny drobnoustrojowe.

Microbiologial studies of the air were performed in two furniture factories. The concentration of microorganisms in the air was low, being of the order 10(3) cfu/m3. The most common organisms were corynebacteria (Arthrobacter, Corynebacterium, Brevibacterium, Microbacterium) and fungi (Aspergillus fumigatus, Rhodotorula rubra). Some of the species found in this environment possess known allergenic properties. Allergological examinations of the workers with environmental aeroallergens have been performed in three departments of one factory. The highest frequency of positive skin reactions were observed among the workers of the varnishing department which may be due to synergistic effects of chemical pollutants. The incidence of precipitin reactions was low among all workers.