Paraseptal Emphysema in Indium Lung: Tracing the Pathological Footprints of Chronic Exposure.

Indium lung is an occupational lung disease caused by exposure to indium-tin-oxide (ITO) dust. Compared to other occupational lung diseases, indium lung has a shorter latency period and the respiratory status continues to worsen even after exposure to the work environment improves. Paraseptal emphysema which affects mainly the subpleural area is seen on chest images obtained via computed tomography (CT), regardless of the smoking history. However, the pathogenesis of emphysema in indium lung is still unclear. Therefore, we re-evaluated the pathology of three previously reported cases of indium lung. Paraseptal emphysema was observed in both smokers and nonsmokers. Obstructive respiratory impairment worsened over time in the cases with paraseptal emphysema. Many alveolar walls were destroyed independent of the presence or absence of emphysetamous changes or fibrosis. Moreover, bronchiolitis was found to be less common in indium lung than in asbestosis (the most common occupational lung disease) or common cases of chronic obstructive pulmonary disease caused by smoking. It has been shown that ITO causes protease anti-protease imbalance, oxidant-antioxidant imbalance, and continuous, abnormal inflammation (the three major causes of emphysema). In addition, nano-sized ITO is less likely to be trapped in the upper airways and may easily reach the subpleural alveoli. Furthermore, ITO may continue to cause sustained tissue injury at the alveolar level potentially resulting in emphysema. Further studies are needed to elucidate the detailed pathogenesis of indium lung by comparing it with other occupational lung diseases.

Biodegradation of polycyclic aromatic hydrocarbons (PAHs) by microbial consortium from paddy rice soil.

Polycyclic aromatic hydrocarbons (PAHs) are among the most widely spread pollutants in the environment including the agricultural soil. PAH degradation by indigenous bacteria is an effective and economical means to remove these pollutants from the environment. Here, we report a bacterial consortium (Pdy-1) isolated from paddy rice soil in northern Japan able to degrade polycyclic aromatic hydrocarbons (PAHs) at high rates. Pdy-1 was incubated with a mixture of PAH compounds (fluorene, phenanthrene, and pyrene) in Bushnell Haas Medium at a final concentration of 100 mg/L each. PDY-1 degraded 100% of fluorene, 95% of phenanthrene, and 52% of pyrene in 5 days. Phenanthrene and pyrene were completely degraded at 10 d and 15 d, respectively. Cloning of the 16S rRNA gene revealed that the consortium was composed of 40% Achromobacter and 7% each of Castelaniella, Rhodanobacter, and Hypomicrobium. Comamonas, Ferrovibrio, Terrimonas, Bordetella, Rhizobium, and Pseudonocardia were also detected. PCR-DGGE showed the dynamics of the consortium during the incubation period. Real-time PCR revealed that PAH degrading genes such as the gram-positive ring dihydroxylating genes (PAH-RDH) and pyrene dioxygenase (nidA) were most abundant at day 5 when the rapid biodegradation of the PAHs was observed. This study improves our understanding on dynamics and characteristics of an effective PAH-degrading bacterial consortium from paddy rice soil.

Metabolic and Epigenetic Regulation of SMAD7 by STC1 Ameliorates Lung Fibrosis.

As shown in our previous studies, the intratracheal-administration of STC1 (stanniocalcin-1) ameliorates pulmonary fibrosis by reducing oxidative and endoplasmic reticulum stress through the uncoupling of respiration in a bleomycin-treated mouse model. However, the overall effect of STC1 on metabolism was not examined. Therefore, we first conducted a comprehensive metabolomics analysis to screen the overall metabolic changes induced by STC1 in an alveolar epithelial cell line using capillary electrophoresis time-of-flight mass spectrometry. The results were subsequently validated in multiple alveolar epithelial and fibroblast cell lines by performing precise analyses of each substance. STC1 stimulated glycolysis, acetyl-CoA synthesis, and the methionine and cysteine-glutathione pathways, which are closely related to the uncoupling of respiration, modulation of epigenetics, and reduction in oxidative stress. These results are consistent with our previous study. Subsequently, we focused on the inhibitory factor SMAD7, which exerts an antifibrotic effect and is susceptible to epigenetic regulation. STC1 upregulates SMAD7 in an uncoupling protein 2-dependent manner, induces demethylation of the SMAD7 promoter region and acetylation of the SMAD7 protein in human alveolar epithelial and fibroblast cell lines and a bleomycin-treated mouse model, and subsequently attenuates fibrosis. The antifibrotic effects of STC1 may partially depend on the regulation of SMAD7. In the evaluation using lung tissue from patients with idiopathic pulmonary fibrosis, SMAD7 expression and acetylation were high in the alveolar structure-preserving region and low in the fibrotic region. The intratracheal administration of STC1 may prevent the development of pulmonary fibrosis by regulating the metabolism-mediated epigenetic modification of SMAD7 in patients.

EphB4 as a Novel Target for the EGFR-Independent Suppressive Effects of Osimertinib on Cell Cycle Progression in Non-Small Cell Lung Cancer.

Osimertinib is the latest generation epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor used for patients with EGFR-mutated non-small cell lung cancer (NSCLC). We aimed to explore the novel mechanisms of osimertinib by particularly focusing on EGFR-independent effects, which have not been well characterized. We explored the EGFR-independent effects of osimertinib on cell proliferation using NSCLC cell lines, an antibody array analysis, and the association between the action of osimertinib and the ephrin receptor B4 (EphB4). We also studied the clinicopathological significance of EphB4 in 84 lung adenocarcinoma patients. Osimertinib exerted significant inhibitory effects on cell growth and cell cycle progression by promoting the phosphorylation of p53 and p21 and decreasing cyclin D1 expression independently of EGFR. EphB4 was significantly suppressed by osimertinib and promoted cell growth and sensitivity to osimertinib. The EphB4 status in carcinoma cells was positively correlated with tumor size, T factor, and Ki-67 labeling index in all patients and was associated with poor relapse-free survival in EGFR mutation-positive patients. EphB4 is associated with the EGFR-independent suppressive effects of osimertinib on cell cycle and with a poor clinical outcome. Osimertinib can exert significant growth inhibitory effects in EGFR-mutated NSCLC patients with a high EphB4 status.

Influence of low temperature on comparative arsenic accumulation and release by three Pteris hyperaccumulators.

Arsenic (As) hyperaccumulator Pteris ferns are renowned for their capacity to accumulate As and have been used to remediate As-contaminated environmental. However, there is less information on how they perform under low temperature though it is important for practical phytoremediation. The purpose of this study was to identify the effect of temperature on As accumulation by three As hyperaccumulators, Pteris multifida, Pteris cretica and Pteris vittata. Ferns were cultured with 5 mg/L As addition under 25 °C to 5 °C for 15 days. The results showed that dropping of temperatures reduced As accumulation by P. vittata moderately but not P. multifida and P. cretica until 10 °C. At 5 °C, all ferns discontinued As accumulation, and the morphology showed necrosis in P. vittata, wherein P. multifida and P. cretica kept healthy. The As distribution showed that As was mainly accumulated in fronds, while P. multifida stored partial As in its root. Both translocation factor and As efflux showed that temperate zone ferns manage As more strictly as compared to P. vittata. Our findings demonstrated that temperature should be considered when applying Pteris ferns for As phytoremediation, and P. multifida could be the most suitable fern for treating As-contaminated water in temperate zone area.

Biodegradation of binary mixtures of octane with benzene, toluene, ethylbenzene or xylene (BTEX): insights on the potential of Burkholderia, Pseudomonas and Cupriavidus isolates.

The contamination of the environment by crude oil and its by-products, mainly composed of aliphatic and aromatic hydrocarbons, is a widespread problem. Biodegradation by bacteria is one of the processes responsible for the removal of these pollutants. This study was conducted to determine the abilities of Burkholderia sp. B5, Cupriavidus sp. B1, Pseudomonas sp. T1, and another Cupriavidus sp. X5 to degrade binary mixtures of octane (representing aliphatic hydrocarbons) with benzene, toluene, ethylbenzene, or xylene (BTEX as aromatic hydrocarbons) at a final concentration of 100 ppm under aerobic conditions. These strains were isolated from an enriched bacterial consortium (Yabase or Y consortium) that prefer to degrade aromatic hydrocarbon over aliphatic hydrocarbons. We found that B5 degraded all BTEX compounds more rapidly than octane. In contrast, B1, T1 and X5 utilized more of octane over BTX compounds. B5 also preferred to use benzene over octane with varying concentrations of up to 200 mg/l. B5 possesses alkane hydroxylase (alkB) and catechol 2,3-dioxygenase (C23D) genes, which are responsible for the degradation of alkanes and aromatic hydrocarbons, respectively. This study strongly supports our notion that Burkholderia played a key role in the preferential degradation of aromatic hydrocarbons over aliphatic hydrocarbons in the previously characterized Y consortium. The preferential degradation of more toxic aromatic hydrocarbons over aliphatics is crucial in risk-based bioremediation.

Leaching of As and Se from coal fly ash: fundamental study for coal fly ash recycling.

Coal fly ash (CFA) is a useful recycled resource for uses such as cement raw material. To manage and evaluate safety for effective utilization of CFA, the leaching concentration and amounts of toxic elements in CFA need to be determined. In this study, 38 types of CFA and aged CFA generated in Japan were used to measure the occurrence and leaching concentration range of As and Se. In addition, the leaching characteristics over the long term were examined using statistical analysis. Leaching concentrations of As and Se from CFAs were in the range of 0.001-0.163 mg/L (average: 0.025 mg/L, median: 0.014 mg/L) and 0.001-0.189 mg/L (average: 0.071 mg/L, median: 0.055 mg/L), respectively. In general, the concentrations of aged CFAs were less than those of the CFAs with a few exceptions. Leaching concentrations of As and Se in the tank leaching test changed with time, and As and Se concentrations in the dispersions increased with stirring time. In contrast, pH of the dispersion decreased with time. The relation between As or Se and CFA factors showed that As or Se and pH or Ca were highly correlated. However, in aged CFAs for long-term use, the correlation coefficient for the relation between As and other factors was low while that for Se-S was high. Considering the effective utilization of CFA as a long-term recyclable resource, the leaching processes of As and Se in CFA would change with time depending on the environmental conditions.

11ß hydroxysteroid dehydrogenase 1: a new marker for predicting response to immune-checkpoint blockade therapy in non-small-cell lung carcinoma.

BACKGROUND: Understanding the status of intratumoural immune microenvironment is necessary to ensure the efficacy of immune-checkpoint (IC) blockade therapy. Cortisol plays pivotal roles in glucocorticoid interactions in the immune system. We examined the correlation between intratumourally synthesised cortisol through 11ß hydroxysteroid dehydrogenase (HSD) 1 and the immune microenvironment in non-small-cell lung carcinoma (NSCLC). METHODS: We correlated 11ßHSD1 immunoreactivity in 125 cases of NSCLC with the amount of intratumoural immune cells present, and 11ßHSD1 immunoreactivity with the efficacy of IC blockade therapy in 18 specimens of NSCLC patients. In vitro studies were performed to validate the immunohistochemical examination. RESULTS: 11ßHSD1 immunoreactivity showed a significant inverse correlation with the number of tumour-infiltrating lymphocytes and CD3- or CD8-positive T cells. 11ßHSD1 immunoreactivity tended to be inversely correlated with the clinical efficacy of the IC blockade therapy. In vitro studies revealed that 11ßHSD1 promoted the intratumoural synthesis of cortisol. This resulted in a decrease in cytokines and in the inhibition of monocyte migration. CONCLUSIONS: Our study is the first report clarifying the inhibitory effects of intratumourally synthesised cortisol through 11ßHSD1 on immune cell migration. We propose that the response to IC blockade therapy in NSCLC may be predicted by 11ßHSD1.

Cupriavidus basilensis strain r507, a toxic arsenic phytoextraction facilitator, potentiates the arsenic accumulation by Pteris vittata.

As a toxic and carcinogenic metalloid, arsenic has posed serious threat to human health. Phytoremediation has emerged as a promising approach to circumvent this problem. Arsenic uptake by Pteris vittata is largely determined by arsenic speciation and mainly occurs via roots; thus, rhizospheric microbial activities may play a key role in arsenic accumulation. The aim of this study was to investigate the potential of arsenic resistant rhizobacteria to enhance arsenic phytoextraction. A total of 49 cultivable rhizobacteria were isolated from the arsenic hyperaccumulating fern, Pteris vittata, and subjected to an initial analysis to identify potentially useful traits for arsenic phytoextraction, such as arsenic resistance and the presence of aioA(aroA)-like (arsenite oxidase-like) gene. Isolated strain r507, named as Cupriavidus basilensis strain r507, was a selected candidate for its outstanding arsenic tolerance, rapid arsenite oxidation ability, and strong colonization to P. vittata. Strain r507 was used in co-cultivation trials with P. vittata in the field for six months. Results showed that the inoculation with strain r507 potentiated As accumulation of P. vittata up to 171%. Molecular analysis confirmed that the inoculation increased the abundance of aioA-like genes in the rhizosphere, which might have facilitated arsenite oxidation and absorption. The findings of this study suggested the feasibility of co-cultivating hyperaccumulators with facilitator bacteria for practical arsenic phytoremediation.

Renal Epithelioid Angiomyolipoma Undergoing Aggressive Clinical Outcome: The MDM2 Expression in Tumor Cells of Two Cases.

Epithelioid angiomyolipoma (EAML) has been known as a potentially malignant tumor which occasionally recur and/or metastasize to other organs, and clinically and pathologically recognized as distinct entity. However, the mechanisms of recurrence and/or metastasis (recurrence/metastasis) has still remained unknown. Here, we report two cases of renal EAML associated with recurrence/metastasis, and three cases of EAML in kidney or liver without recurrence/metastasis. According to the previous histological predictive models of EAML, the primary tumor was classified as low risk group in one of the cases with recurrence/metastasis in spite of its malignant behavior. Therefore, we considered that further investigation about the mechanisms of recurrence/metastasis in EAML is required for a malignancy prediction. We focused on some cell-cycle modulators, including mouse double minute 2 homolog (MDM2), which is ubiquitin ligase well-known to promote malignant behaviors by p53 ubiquitination and degradation, and also other cellular processes including genomic instability and epithelial-mesenchymal transition in p53-independent manners in various human malignancies. Immunohistochemical evaluation revealed that MDM2 protein expression increased stepwise throughout every steps of metastasis/recurrence in both cases, although it was negative in primary tumors. In conclusion, this is the first study demonstrating that MDM2 could play an important role in the molecular mechanisms of recurrence/metastasis of EAML. Further analyses focusing on MDM2 pathway could contribute to the identification of novel prognostic factors and/or therapeutic targets in EAML patients.

Classification of coal fly ash based on pH, CaO content, glassy components, and leachability of toxic elements.

Coal fly ash (CFA), a by-product generated from coal-burning power plants, readily leaches toxic elements into aquatic environments. The present study describes a classification system for CFA based on the chemical composition of CFA and leachability of toxic elements, which can promote the safe and effective utilization of CFA for uses such as fly ash cement. To classify CFA samples, the CaO content, leachate pH, leachability of toxic elements such as B, As, and Se, and the acid- and alkali-soluble Si and Al in glassy components were determined for ten types of CFA samples produced in Japan. The results indicated that the CFA samples could be grouped into three groups: group A, which was characterized by low CaO content, low leachate pH, and a relatively high amount of alkaline-soluble Al; group B, which was characterized by low CaO content, low leachate pH, and relatively low amount of alkaline-soluble Al; and group C, which was characterized by high CaO content, high leachate pH, and relatively low amount of alkaline-soluble Al. Characteristic of group A CFA was the simultaneous leaching of Al and the minor elements along with a gradual increase in pH. This type of CFA carries the risk of leaching toxic substances upon contact with alkali solutions. These results can aid the discovery and separation of safe and unsafe CFA, allowing the safe CFA to be used in cement to produce concrete under alkali conditions.

Arsenic, lead and cadmium removal potential of Pteris multifida from contaminated water and soil.

The main threats to the environment from heavy metals are associated with arsenic (As), lead (Pb) and cadmium (Cd). In this study, the potential of Pteris multifida for removing As, Pb and Cd from hydroponic solution and pot soil was evaluated for the first time. Short-term (5 day) experiments were conducted to assess phytofiltration efficiency of temperate zone fern P. multifida and to compare it with mostly studied tropical zone fern P. vittata. Within 5 days, P. multifida accumulated 33% of As(III), whereas P. vittata could not accumulate that most toxic arsenic species As(III) at all. Long-term hydroponic results showed that 90% of Pb, 50% of As and 36% of Cd were removed by P. multifida. Concentration of As in the frond (22 mg/kg dw) was comparatively higher than other parts of plant and significantly higher concentration of Cd and Pb were stored in root and rhizome. Pot soil experiment of P multifida confirmed the comparative uptake and translocation of As(V), Pb and Cd from soil. Therefore, from the assessment of heavy metal accumulation capacity, translocation and healthy survival for long time, P. multifida was identified as an excellent species for the treatment of multi-metal contaminated water and soil.

The Significance of MMP-1 in EGFR-TKI-Resistant Lung Adenocarcinoma: Potential for Therapeutic Targeting.

Epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) resistance is one of the most important problems in lung cancer therapy. Lung adenocarcinoma with EGFR-TKI resistance was reported to have higher abilities of invasion and migration than cancers sensitive to EGFR-TKI, but the function of matrix metalloproteinases (MMPs) has not been explored in EGFR-TKI-resistant lung adenocarcinoma. This study aims to clarify the significance of MMP-1 in EGFR-TKI-resistant lung adenocarcinoma. From the results of in vitro studies of migration and invasion assays using EGFR-TKI-sensitive and -resistant cell lines and phosphorylation antibody arrays using EGF and rapamycin, we first demonstrate that overexpression of MMP-1, which might follow activation of a mammalian target of rapamycin (mTOR) pathway, plays an important role in the migration and invasion abilities of EGFR-TKI-resistant lung adenocarcinoma. Additionally, immunohistochemical studies using 89 cases of lung adenocarcinoma demonstrate that high expression of MMP-1 is significantly correlated with poor prognosis and factors such as smoking history and the subtype of invasive mucinous adenocarcinoma. These are consistent with the results of this in vitro study. To conclude, this study provides insights into the development of a possible alternative therapy manipulating MMP-1 and the mTOR signaling pathway in EGFR-TKI-resistant lung adenocarcinoma.

Direct Self-Sustained Fragmentation Cascade of Reactive Droplets.

A traditional hand-held firework generates light streaks similar to branched pine needles, with ever smaller ramifications. These streaks are the trajectories of incandescent reactive liquid droplets bursting from a melted powder. We have uncovered the detailed sequence of events, which involve a chemical reaction with the oxygen of air, thermal decomposition of metastable compounds in the melt, gas bubble nucleation and bursting, liquid ligaments and droplets formation, all occurring in a sequential fashion. We have also evidenced a rare instance in nature of a spontaneous fragmentation process involving a direct cascade from big to smaller droplets. Here, the self-sustained direct cascade is shown to proceed over up to eight generations, with well-defined time and length scales, thus answering a century old question, and enriching, with a new example, the phenomenology of comminution.

Analysis of stable 1,2-dichlorobenzene-degrading enrichments and two newly isolated degrading strains, Acidovorax sp. sk40 and Ralstonia sp. sk41.

Stable degrading 1,2-dichlorobenzene (1,2-DCB) enrichments were generated from original contaminated soil and groundwater via enrichment procedures using a mineral salt medium containing 1,2-DCB as the sole carbon and energy source. Four transferred enrichments showed stable 1,2-DCB-degrading ability and completely degraded 1,2-DCB within 32 h. PCR-denaturing gradient gel electrophoresis (DGGE) and 16S rRNA gene clone library analyses indicated that two bacterial strains, belonging to Acidovorax spp. and Ralstonia spp., respectively, were the predominant organisms in each enrichment. Moreover, these strains maintained a stable coexistence in the four transferred enrichments. These two bacteria were subsequently identified as Acidovorax sp. strain sk40 and Ralstonia sp. strain sk41. Strain sk40 was more tolerant to higher concentrations of 1,2-DCB than strain sk41, while strain sk41 maintained a shorter degradation time under lower concentrations of 1,2-DCB. Notably, however, both strains exhibited similar growth rates and degradation rates in media containing 40 mg/l 1,2-DCB, as well as complete degradation of the 1,2-DCB (40 mg/l) within 32 h. It is expected that these two strains will be used in future applications of bioremediation of 1,2-DCB contamination.

Development of suitable hydroponics system for phytoremediation of arsenic-contaminated water using an arsenic hyperaccumulator plant Pteris vittata.

In this study, we found that high-performance hydroponics of arsenic hyperaccumulator fern Pteris vittata is possible without any mechanical aeration system, if rhizomes of the ferns are kept over the water surface level. It was also found that very low-nutrition condition is better for root elongation of P. vittata that is an important factor of the arsenic removal from contaminated water. By the non-aeration and low-nutrition hydroponics for four months, roots of P. vittata were elongated more than 500 mm. The results of arsenate phytofiltration experiments showed that arsenic concentrations in water declined from the initial concentrations (50 µg/L, 500 µg/L, and 1000 µg/L) to lower than the detection limit (0.1 µg/L) and about 80% of arsenic removed was accumulated in the fern fronds. The improved hydroponics method for P. vittata developed in this study enables low-cost phytoremediation of arsenic-contaminated water and high-affinity removal of arsenic from water.

Evaluation of the effectiveness and salt stress of Pteris vittata in the remediation of arsenic contamination caused by tsunami sediments.

On March 11, 2011, one of the negative effects of the tsunami phenomenon that devastated the Pacific coast of the Tohoku district in Japan was the deposition of a wide range of arsenic (As) contamination to the soil. To remediate such a huge area of contamination, phytoremediation by Pteris vittata, an As-hyperaccumulator, was considered. To evaluate the efficacy of applying P. vittata to the area, the salt tolerance of P. vittata and the phytoextraction of As from soil samples were investigated. For the salt tolerance test, spore germination was considerably decreased at an NaCl level of more than 100 mM. At 200 mM, the gametophytes exhibited a morphological defect. Furthermore, the growth inhibition of P. vittata was observed with a salinity that corresponded to 66.2 mS/m of electric conductivity (EC) in the soil. A laboratory phytoremediation experiment was conducted using As-contaminated soils for 166 days. P. vittata grew and accumulated As at 264 mg/kg-DW into the shoots. Consequently, the soluble As in the soil was evidently decreased. These results showed that P. vittata was applicable to the phytoremediation of As-contaminated soil with low salinity as with the contamination caused by the 2011 tsunami.

Immobilization of B, F, Cr, and As in alkaline coal fly ash through an aging process with water.

Fourteen different alkaline coal fly ashes (CFAs) were used for the experiment, in which each sample was mixed with water to be 28.6% of water content (wt/wt) and aged for 1-4 weeks at 10-30 °C. This simple treatment is advantageous for decreases in water-soluble B, F, Cr, and As. Compared to non-aged CFAs, their water-soluble fractions remained 0.56-88%, 21-85%, 0.37-93% and 2.6-88%, respectively, after aging for a week at 20 °C, although the amounts of Cr and As released from some CFA samples increased. Considering the significant decrease in elution of sulfate, Ca and Al after aging, the immobilization, namely prevention of toxic element elution, could be related to formation of secondary minerals such as portlandite, gypsum and ettringite. Immobilization of B and Cr tends to proceed preferentially under colder conditions. Aging at higher temperatures enhances the leachability of Cr in some CFA samples. Contrary to the behavior of B and Cr, water-soluble F effectively decreases under warmer conditions.