Silica-carrying particulate matter enhances Bjerkandera adusta-induced murine lung eosinophilia.

Bjerkandera adusta (B. adusta) causes fungus-associated chronic cough. However, the inflammatory response is not yet fully understood. Recently, B. adusta was identified in Asian sand dust (ASD) aerosol. This study investigated the enhancing effects of ASD on B. adusta-induced lung inflammation. B. adusta was inactivated by formalin. ASD was heated to remove toxic organic substances. ICR mice were intratracheally instilled with saline, B. adusta 0.2 µg, or B. adusta 0.8 µg with or without heated ASD 0.1 mg (H-ASD), four times at 2-week intervals. Two in vitro experiments were conducted to investigate any enhancing effects using bone marrow-derived macrophages (BMDM) from Toll-like receptor (TLR) knockout mice and ICR mice. Co-exposure to H-ASD and B. adusta, especially at high doses, caused eosinophil infiltration, proliferation of goblet cells in the airway, and fibrous thickening of the subepithelial layer, and remarkable increases in expression of Th2 cytokines and eosinophil-related cytokine and chemokine expression in bronchoalveolar lavage fluid. In the in vitro study using BMDM from wild-type, TLR2-/-, and TLR4-/- mice, the TLR-signaling pathway for cytokine production caused by B. adusta was predominantly TLR2 rather than TLR4. H-ASD increased the expression of NF-κB and cytokine production by B. adusta in BMDM from ICR mice. The results suggest that co-exposure to H-ASD and B. adusta caused aggravated lung eosinophilia via remarkable increases of pro-inflammatory mediators. The aggravation of inflammation may be related, at least in part, to the activation of the TLR2-NF-κB signaling pathway in antigen presenting cells by H-ASD.

The Role of Toll-Like Receptors and Myeloid Differentiation Factor 88 in Bjerkandera adusta-Induced Lung Inflammation.

BACKGROUND: Recently, a cluster of patients with an intractable allergic fungal cough who were characterized by sensitization to Bjerkandera adusta was reported. In the present study, the role of Toll-like receptors and myeloid differentiation factor 88 (MyD88) in B. adusta-induced lung inflammation was investigated. METHODS: Wild-type (WT), TLR2-/-,TLR4-/-, and MyD88-/- BALB/c mice were intratracheally challenged with B. adusta 4 times at 2-week intervals. Lung pathology, bronchoalveolar lavage fluid (BALF) cytological profiles, and inflammatory mediators in BALF were investigated. Bone marrow-derived macrophages (BMDM) from TLR2-/-,TLR4-/-, TLR2/4-/-, TLR7/9-/-,MyD88-/-, and WT C57BL/6J mice were stimulated with B. adusta for 12 h, and inflammatory mediators in the culture medium were measured. RESULTS: B. adusta caused lung inflammation along with Th2 cytokine [interleukin (IL)-5 and IL-13] and eosinophil-related chemokine [eotaxin and monocyte chemotactic protein (MCP-3)] production, an increase in eosinophils in BALF, and eosinophil infiltration in the airways in WT and TLR4-/- mice. However, Th2 and eosinophil-related responses in TLR2-/- and MyD88-/- mice were low or undetectable. The induction of neutrophils and IL-6, IL-12, IL-17A, and MCP-1 in the BALF of MyD88-/- mice was attenuated compared to that in WT mice. The induction of IL-6, TNF-α, MCP-1, and macrophage inflammatory protein-1α was reduced or undetectable in B. adusta-stimulated BMDM from TLR7/9-/- and MyD88-/- mice compared to WT mice. CONCLUSIONS: These results suggest that TLR2 and the adapter protein MyD88 may play an important role in the induction of eosinophils by B. adusta. However, TLR7/9-MyD88 might be important in the induction of neutrophils and the relevant inflammatory mediators, especially IL-17A.

Function of rayon fibers with metallophthalocyanine derivatives: potential of low-molecular weight polycyclic aromatic hydrocarbon removal and Bacillus sp. removal.

Polycyclic aromatic hydrocarbons (PAHs) are known as carcinogenic and/or mutagenic substances, and are present at high concentration in polluted environments. It has recently been reported that spore-forming bacteria (e.g., Bacillus spp.) can be transported long distances alive in the atmosphere, which raises the possibility that some of the transported bacteria could have adverse effects on human health. There is thus a need for filters that can remove gaseous PAHs from the air that people breathe and that can inhibit bacterial growth on the filters. We focused on metallophthalocyanine derivatives (M-Pc) which are known to adsorb PAHs as well as to inhibit the growth of bacteria as a potential filtering agent. In this study, we developed different types of M-Pc-supported rayon fibers by changing central metals, functional groups, concentrations of M-Pc and rayon types, and evaluated their removal effects by measuring adsorption rates of 3- and 4-ring PAHs with a HPLC and growth curves of Bacillus sp. with a spectrophotometer. The results showed that both the effects depended on functional groups and concentrations of M-Pc, and rayon types. The most effective combination was observed in Fe-Pc with sulfo group supported on cationized rayon fiber at the concentration of 2 to 3.3 wt%. Central metal species of M-Pc were influenced only on the antibacterial properties. This fiber would be applicable to filtering agents and textiles.

Bioprocess of Kosa bioaerosols: effect of ultraviolet radiation on airborne bacteria within Kosa (Asian dust).

Kosa (Asian dust) is a well-known weather phenomenon in which aerosols are carried by the westerly winds from inland China to East Asia. Recently, the frequency of this phenomenon and the extent of damage caused have been increasing. The airborne bacteria within Kosa are called Kosa bioaerosols. Kosa bioaerosols have affected ecosystems, human health and agricultural productivity in downwind areas. In order to develop a new and useful bacterial source and to identify the source region of Kosa bioaerosols, sampling, isolation, identification, measurement of ultraviolet (UV) radiation tolerance and experimental simulation of UV radiation conditions were performed during Kosa bioaerosol transportation. We sampled these bioaerosols using a Cessna 404 airplane and a bioaerosol sampler at an altitude of approximately 2900 m over the Noto Peninsula on March 27, 2010. The bioaerosol particles were isolated and identified as Bacillus sp. BASZHR 1001. The results of the UV irradiation experiment showed that the UV radiation tolerance of Kosa bioaerosol bacteria was very high compared with that of a soil bacterium. Moreover, the UV radiation tolerance of Kosa bioaerosol spores was higher than that of soil bacterial spores. This suggested that Kosa bioaerosols are transported across the atmosphere as living spores. Similarly, by the experimental simulation of UV radiation conditions, the limited source region of this Kosa bioaerosol was found to be southern Russia and there was a possibility of transport from the Kosa source area.

Variations in the structure of airborne bacterial communities in a downwind area during an Asian dust (Kosa) event.

Asian dust (Kosa) events transport airborne microorganisms that significantly impact biological ecosystems, human health, and ice-cloud formation in downwind areas. However, the composition and population dynamics of airborne bacteria have rarely been investigated in downwind areas during Kosa events. In this study, air samplings were sequentially performed at the top of a 10-m high building within the Kosa event arrival area (Kanazawa City, Japan) from May 1 to May 7, 2011, during a Kosa event. The particle concentrations of bacterial cells and mineral particles were ten-fold higher during the Kosa event than on non-Kosa event days. A 16S ribosomal DNA clone library prepared from the air samples primarily contained sequences from three phyla: Cyanobacteria, Firmicutes, and Alphaproteobacteria. The clones from Cyanobacteria were mainly from a marine type of Synechococcus species that was dominant during the first phase of the Kosa event and was continuously detected throughout the Kosa event. The clones from Alphaproteobacteria were mainly detected at the initial and final periods of the Kosa event, and phylogenetic analysis showed that their sequences clustered with those from a marine bacterial clade (the SAR clade) and Sphingomonas spp. During the middle of the Kosa event, the Firmicutes species Bacillus subtilis and Bacillus pumilus were predominant; these species are known to be predominant in the atmosphere above the Chinese desert, which is the source of the dust during Kosa events. The clones obtained after the Kosa event had finished were mainly from Bacillus megaterium, which is thought to originate from local terrestrial areas. Our results suggest that airborne bacterial communities at the ground level in areas affected by Kosa events change their species compositions during a Kosa event toward those containing terrestrial and pelagic bacteria transported from the Sea of Japan and the continental area of China by the Kosa event.

Lung inflammation by fungus, Bjerkandera adusta isolated from Asian sand dust (ASD) aerosol and enhancement of ovalbumin-induced lung eosinophilia by ASD and the fungus in mice.

BACKGROUND: Bjerkandera adusta (B. adusta) is one of the most important etiological fungi associated with chronic cough. However, precise details of the inflammatory response to exposure are not well understood yet. B. adusta was recently identified in Asian sand dust (ASD) aerosol. Therefore, in the present study the exacerbating effects of ASD on B. adusta-induced lung inflammation and B. adusta + ASD on ovalbumin (OVA)-induced murine lung eosinophilia were investigated using experimental mice. METHODS: In order to prepare testing samples, B. adusta obtained from ASD aerosol was inactivated by formalin and ASD collected from the atmosphere was heated to remove toxic organic substances (H-ASD). CD-1 mice were instilled intratracheally with 12 different samples prepared with various combinations of B. adusta, H-ASD, and OVA in a normal saline solution. The lung pathology, cytological profiles in bronchoalveolar lavage fluid (BALF), and the levels of inflammatory cytokines/chemokines in BALF were investigated. RESULTS: H-ASD aggravated the lung eosinophilia induced by B. adusta alone, which also aggravated the lung eosinophilia induced by OVA. The mixture of OVA, H-ASD, and B. adusta caused serious fibrous thickening of the subepithelial layer, eosinophil infiltration, and proliferation of goblet cells in the airways along with remarkable increases of IL-13, eotaxin, IL-5, and MCP-3 in BALF. CONCLUSIONS: The results of the present study demonstrated that B. adusta isolated from ASD aerosol induces allergic lung diseases. H-ASD enhanced allergic reactions caused by OVA or B. adusta. A mixture of B. adusta, H-ASD, and OVA caused the most remarkable exacerbation to the allergic airway inflammation via remarkable increases of pro-inflammatory mediators.

The utilization of Triton X-100 for enhanced two-dimensional liquid-phase proteomics.

One of the main challenges in proteomics lies in obtaining a high level of reproducible fractionation of the protein samples. Automated two-dimensional liquid phase fractionation (PF2D) system manufactured by Beckman Coulter provides a process well suited for proteome studies. However, the protein recovery efficiency of such system is low when a protocol recommended by the manufacturer is used for metaproteome profiling of environmental sample. In search of an alternative method that can overcome existing limitations, this study replaced manufacturer's buffers with Triton X-100 during the PF2D evaluation of Escherichia coli K12. Three different Triton X-100 concentrations-0.1%, 0.15%, and 0.2%-were used for the first-dimension protein profiling. As the first-dimension result was at its best in the presence of 0.15% Triton X-100, second-dimension protein fractionation was performed using 0.15% Triton X-100 and the standard buffers. When 0.15% Triton X-100 was used, protein recovery increased as much as tenfold. The elution reliability of 0.15% Triton X-100 determined with ribonuclease A, insulin, α-lactalbumin, trypsin inhibitor, and cholecystokinin (CCK) affirmed Triton X-100 at 15% can outperform the standard buffers without having adverse effects on samples. This novel use of 0.15% Triton X-100 for PF2D can lead to greater research possibilities in the field of proteomics.

Phylogenetic analysis of atmospheric halotolerant bacterial communities at high altitude in an Asian dust (KOSA) arrival region, Suzu City.

The microbial communities transported by Asian desert dust (KOSA) events have attracted much attention as bioaerosols because the transported microorganisms are thought to influence the downwind ecosystems in Korea and Japan. However, the atmospheric microbial community has not been investigated at high altitude in the KOSA arrival area. In this study, to estimate the viability and diversity of atmospheric halotolerant bacteria, which are expected to resist to various environmental stresses as well as high salinities, bioaerosol samples were collected at 10 and 600 m above the ground within the KOSA arrival area, Suzu City, Japan, during KOSA events. During the sampling period, the particle numbers at 600 m were higher than those at 10 m, suggesting that large particles of aerosol fall from the high altitude of 600 m to the ground surface. The microorganisms in bioaerosol samples grew in media containing up to 15% NaCl concentrations demonstrating the viability of the halotolerant bacteria in bioaerosol samples. The PCR-DGGE analysis using 16S rDNA revealed that the bacterial species in NaCl-amended cultures were similar to the bacteria detected from the genomic DNA directly extracted from the bioaerosol samples. The 16S rDNA sequences of bacterial communities in bioaerosol samples were classified into 4 phylotypes belonging to the Bacilluscereus or Bacillussubtilis group. The bioaerosol samples collected at 600 m included 2 phylotypes belonging to B. subtilis, and one phylotype among all 4 phylotypes was identical between the samples at 10 and 600 m. In the atmosphere at 600 m, the halotolerant bacterial community was expected to remain viable, and the species composition was expected to include a few species of the genus Bacillus. During this investigation period, these atmospheric bacteria may have been vertically transported to the ground surface, where the long-range KOSA particle transport from China is frequently observed.

Immobilization of cross-linked lipase aggregates onto magnetic beads for enzymatic degradation of polycaprolactone.

Candida rugosa lipase was immobilized on amino-functionalized magnetic supports via cross-linked enzyme aggregates (CLEA) and used to enhance the enzymatic degradation of polycaprolactone (PCL). The maximum amounts of lipase immobilized on the magnetic beads using glutaraldehyde as a coupling agent were determined to be 33.7 mg/g of beads with an 81% recovery of activity after immobilization. Compared to the free enzyme, the immobilized lipase showed the optimum pH at 1 unit higher (pH 8.0) and also retained its enzymatic activity at higher temperatures. There was 62.9% retention of lipase activity after 30 consecutive reuses, indicating its stability and reusability in aqueous media. Moreover, the immobilized lipase maintained more than 80% of its initial activity during 30 days storage period, while the free lipase lost all under same condition. In addition, the immobilized lipase showed a more than 6-fold increase in biodegradability over the free lipase when the immobilized lipase was used to degrade PCL in a batch system. Higher thermal and storage stability, as well as good durability after repeated use of the immobilized lipase CLEA, highlights its potential applicability as large scale continuous systems for the enzymatic degradation of PCL.

Biodegradation of diisodecyl phthalate (DIDP) by Bacillus sp. SB-007.

In this study, diisodecyl phthalate (DIDP) was efficiently degraded by Bacillus sp. SB-007. The optimal conditions for DIDP (100 mg l(-1)) degradation by Bacillus sp. SB-007 in a mineral salts medium were found to be pH 7.0 at 30 degrees C, stirring at 200 rpm. The specific rate of DIDP degradation was found to be concentration dependent with a maximum of 4.87 mg DIDP l(-1) h(-1). DIDP was transformed rapidly by Bacillus sp. SB-007 with the formation of monoisodecyl phthalate and phthalic acid, which subsequently degraded further. These results highlight the potential of this bacterium for removing DIDP contaminated waste in the environment.

Purification of seawater contaminated with undegradable aromatic ring compounds using ozonolysis followed by titanium dioxide treatment.

Treatment of aromatic ring compounds, 2,4-dichlorophenoxy acetic acid (2,4-D), 2,4,5-trichloro-phenoxy acetic acid (2,4,5-T), and bisphenol A, in the artificial seawater, i.e. Allen seawater, was carried out by ozonation and titanium dioxide (TiO2) photocatalyst treatment. Each compound was degraded and varnished within 30 min by only ozonolysis at pH 9.0 and at 20 degrees C, while the TOC value of each compound decreased gradually but reached almost constant value, i.e. about 70-80% of the initial value, at even 30 min of ozonation time. Ozonolysis (30 min of ozonation time) followed by TiO2 photocatalyst treatment (50h of reaction time) was a very effective method for decreasing the TOC values of aromatic ring compounds in the artificial seawater. In consequence, TOC values of 2,4-D, 2,4,5-T, and bisphenol A could be reduced to about 28, 21, and 34% of their initial values, respectively.

Determination of Cu, Pb, Fe, and Zn in plant component polymers of a hyperaccumulating plant.

Phytoremediation is an innovative technology that utilizes the natural properties of plants to remediate hazardous waste sites. For more cost-effective phytoremediation, it is important to utilize a hyperaccumulating plant after phytoremediation, i.e. the recovery of valuable metals and the production of useful materials. In this work, the determination of metals in plant component polymers in a fern, Athyrium yokoscense, as a hyper-accumulating plant was established using steam explosion, Wayman's extraction method, and ICP emission spectrometry. After A. yokoscense plants were treated by steam explosion, the steam-exploded A. yokoscense were separated into four plant component polymers, ie. water-soluble material fraction, holocellulose fraction, methanol-soluble lignin fraction, and residual lignin fraction. The concentrations of Cu, Pb, Fe, and Zn in these plant component polymers and the dry weights of plant component polymers were measured. These analytical process determining metals in the plants will contribute to not only the evaluation and the efforts of phytoremediation using a hyperaccumulating plant, but also to the development of more effective phytoremediation.

Methane production from steam-exploded bamboo.

To convert unutilized plant biomass into a useful energy source, methane production from bamboo was investigated using a steam explosion pretreatment. Methane could not be produced from raw bamboo but methane production was enhanced by steam explosion. The maximum amount of methane produced, i.e., about 215 ml, was obtained from 1 g of exploded bamboo at a steam pressure of 3.53 MPa and a steaming time of 5 min. A negative correlation between the amount of methane produced and the amount of Klason lignin was observed in the methane fermentation of steam-exploded bamboo.

Efficient production by Escherichia coli of recombinant protein using salting-out effect protecting against proteolytic degradation.

To produce glucoamylase efficiently as a recombinant protein, E. coli was grown with 20 g (NH4)2SO4 l-1 which removed proteolytic activity but did not effect cell growth. Growth in M9 medium with 20 g (NH4)2SO4 l-1 produced 11 U glucoamylase ml-1 compared to 7 U ml-1 without addition. Furthermore, the glucoamylase activity was maintained at about 9 U ml-1.