Lemborexant-induced interstitial lung disease: A case report.

We report the first case of drug-induced interstitial lung disease attributed to lemborexant. A 66-year-old man reported to our hospital with the acute onset of cough and breathlessness with ground-glass opacity on radiological examination. Symptoms were identified after taking lemborexant for 2 consecutive days. The patient had undergone lemborexant treatment 2 years prior and had exhibited no symptoms at that time. The drug-induced lymphocyte stimulation test for lemborexant was positive. He showed rapid improvement upon treatment with steroid. With the rise in prescriptions of lemborexant for insomnia, lemborexant should be considered as a possible cause of drug-induced interstitial lung disease.

Influential factors of long-term and seasonal 137Cs change in agricultural and forested rivers: Temperature, water quality and an intense Typhoon Event.

In this study, the effect of temperature, water quality, and the impact of an intense typhoon event on change in 137Cs concentration in the water of agricultural and forested rivers near the Fukushima Daiichi nuclear power plant (Japan) was evaluated using monthly stationary observations obtained under baseflow conditions 2.8-10.6 years after the nuclear accident in 2011. The dissolved 137Cs concentration fluctuated seasonally with water temperature in all rivers, and the increase in dissolved 137Cs concentration for unit increase in temperature was higher in forested rivers than in agricultural rivers. The relationship between water temperature and the apparent distribution coefficient of 137Cs well followed the van 't Hoff equation in the two agricultural rivers, where the enthalpy of reaction was estimated as -15.6 and -19.6 kJ mol-1. The van 't Hoff equation was not well followed for a forested river, where the suspended solids mainly comprised organic matter, suggesting that the dominant process determining dissolved 137Cs concentrations in forested rivers is not only water temperature effect on ion exchange, but rather the input of 137Cs and K+ (competing with 137Cs for exchange sites on mineral particles) into the water phase via litter leaching. Suspended solids concentrations in agricultural rivers correlated negatively with 137Cs concentrations in suspended solids, suggesting an increased proportion of coarse particles or the input of soils with low 137Cs concentration from decontaminated agricultural land. At some sites, 137Cs concentrations in dissolved form and in suspended solids were reduced sharply in association with the passage of Typhoon Hagibis in October 2019, suggesting that Typhoon Hagibis caused large-scale surface erosion that removed the source of 137Cs.

Regulation of PD-L1 expression in non-small cell lung cancer by interleukin-1ß.

Introduction: Programmed cell death-ligand 1 (PD-L1) is a biomarker for prediction of the clinical efficacy of immune checkpoint inhibitors in various cancer types. The role of cytokines in regulation of PD-L1 expression in tumor cells has not been fully characterized, however. Here we show that interleukin-1ß (IL-1ß) plays a key role in regulation of PD-L1 expression in non-small cell lung cancer (NSCLC). Methods: We performed comprehensive screening of cytokine gene expression in NSCLC tissue using available single-cell RNA-Sequence data. Then we examined the role of IL-1ß in vitro to elucidate its induction of PD-L1 on NSCLC cells. Results: The IL-1ß gene is highly expressed in the tumor microenvironment, particularly in macrophages. The combination of IL-1ß and interferon-γ (IFN-γ) induced a synergistic increase in PD-L1 expression in NSCLC cell lines. IL-1ß and IFN-γ also cooperatively activated mitogen-activated protein kinase (MAPK) signaling and promoted the binding of downstream transcription factors to the PD-L1 gene promoter. Furthermore, inhibitors of MAPK signaling blocked upregulation of PD-L1 by IL-1ß and IFN-γ. Discussion: Our study reports high levels of IL-1ß in the tumor microenvironment may cooperate with IFN-γ to induce maximal PD-L1 expression in tumor cells via activation of MAPK signaling, with the IL-1ß-MAPK axis being a promising therapeutic target for attenuation of PD-L1-mediated suppression of antitumor immunity.

Contrasting radiocesium transfer in the river and lake food webs: Importance of trophic level and food source.

Radiocesium (137Cs) contamination of the freshwater ecosystems adjacent to the Fukushima Daiichi Nuclear Power Plant (FDNPP) in Japan has persisted long after the accident that occurred at the facility in March 2011. It is necessary to elucidate the dynamics of 137Cs in various aquatic ecosystems to predict 137Cs concentrations in fish and manage freshwater fisheries in the vicinity of FDNPP. To these ends, we applied stable isotope analysis to evaluate changes in 137Cs levels through trophic positions and the relative importance of the 137Cs sources at the trophic bases of two rivers and two lakes in Fukushima. The δ15N analyses disclosed that 137Cs decreases from primary producers to fish consumers in the river food web and 137Cs increases among fish consumers with increasing trophic position in the lake food web. The δ13C analysis revealed that autochthonous 137Cs contributed to fish contamination. The periphyton-dependent and zooplankton-dependent fish had comparatively higher 137Cs concentrations in the rivers and lakes, respectively. Cesium-137 supply from the pelagic food web was observed to contribute to greater 137Cs levels in the fish consumers inhabiting the lakes. The results of this study show that stable isotope analysis may help clarify 137Cs dynamics in freshwater food webs and identify the important 137Cs sources in the food web. Identifying important 137Cs sources and trophic transfers depending on the ecosystem help guide regulatory and management frameworks to establish profitability of the food fish stocks there and maintain food security.

Contrasting seasonality of 137Cs concentrations in two stream animals that share a trophic niche.

Understanding the seasonality of 137Cs concentrations in aquatic animals is crucial for reviving local inland fisheries. The seasonality of 137Cs concentrations in animals is expected to vary, even if focal species consume similarly contaminated foods because the 137Cs excretion rate is species-specific, and 137Cs uptake by foraging autochthonous food resources also vary among seasons. Here, we conducted a seasonal monitoring survey of dissolved 137Cs concentrations as an indicator of the contamination level of food resources and measured 137Cs concentrations in two carnivorous aquatic animals (Palaemon paucidens and Rhinogobius sp.) that share a trophic niche in a stream connected to a dam reservoir. The dissolved 137Cs concentration had clear seasonality-high in summer and low in winter. The 137Cs concentrations in the animals revealed a different seasonal pattern-it peaked in October in P. paucidens and peaked in February in Rhinogobius. Overall, the 137Cs concentration was relatively higher in P. paucidens than in Rhinogobius, suggesting that P. paucidens has a lower excretion rate than Rhinogobius. Consequently, the seasonality of the 137Cs concentration in P. paucidens showed temporal changes similar to those of the dissolved 137Cs concentration, which were likely affected by 137Cs uptake through foraging, whereas that in Rhinogobius was controlled by 137Cs excretion. This study shows that the seasonality of 137Cs concentration can differ between sympatric animals that share a trophic niche. Accumulating knowledge and comparing the seasonality of 137Cs concentrations in fisheries species based on the balance between uptake and excretion will be valuable to determine the appropriate seasons to obtain less-contaminated products.

Radiocesium-bearing microparticles cause a large variation in 137Cs activity concentration in the aquatic insect Stenopsyche marmorata (Tricoptera: Stenopsychidae) in the Ota River, Fukushima, Japan.

After the Tokyo Electric Power Company Fukushima Daiichi Nuclear Power Plant accident in Japan, freshwater ecosystems near the site remained contaminated by radiocesium (RCs). Clarifying RCs concentrations in aquatic insects is crucial because fishes consume these insects that transfer RCs into freshwater ecosystems. As aquatic insects are usually measured for radioactivity in bulk samples of several tens of insects, variation in RCs concentration among individuals is not captured. In this study, we investigated the variability in 137Cs activity concentration in individual aquatic insects in detritivorous caddisfly (Stenopsyche marmorata) and carnivorous dobsonfly (Protohermes grandis) larvae from the Ota River, Fukushima. Caddisfly larvae showed sporadically higher radioactivity in 4 of the 46 caddisfly larvae, whereas no such outliers were observed in 45 dobsonfly larvae. Autoradiography and scanning electron microscopy analyses confirmed that these caddisfly larvae samples contained radiocesium-bearing microparticles (CsMPs), which are insoluble Cs-bearing silicate glass particles. CsMPs were also found in potential food sources of caddisfly larvae, such as periphyton and drifting particulate organic matter, indicating that larvae may ingest CsMPs along with food particles of similar size. Although CsMP distribution and uptake by organisms in freshwater ecosystems is relatively unknown, our study demonstrates that CsMPs can be taken up by aquatic insects.

Inhibition of PI3Kδ Differentially Regulates Poly I:C- and Human Metapneumovirus-Induced PD-L1 and PD-L2 Expression in Human Bronchial Epithelial Cells.

Bronchial epithelial cells are front sentinels eliciting innate and adaptive immunity to respiratory viral pathogens. Recognition of viral double-stranded RNA induces antiviral interferon (IFN) responses in bronchial epithelial cells. Co-inhibitory molecules programmed cell death 1 ligand 1 (PD-L1) and ligand 2 (PD-L2) were also induced on bronchial epithelial cells, which bind programmed cell death 1 on T cell and inhibit the function of virus-specific cytotoxic T lymphocyte. A previous study showed that antiviral type I IFN increased PD-L1 and PD-L2 expression in cultured melanoma cells. However, it remains unknown whether antiviral IFNs affect PD-L1 and PD-L2 expression in bronchial epithelial cells. In addition, we previously reported that inhibition of PI3Kδ signaling enhanced antiviral IFN responses in human primary bronchial epithelial cells (PBECs). Here we assessed the effect of exogenous IFNs or a selective PI3Kδ inhibitor IC87114 on PD-L1 and PD-L2 in PBECs stimulated with a synthetic double-stranded RNA poly I:C or human metapneumovirus. Treatment with IFNß or IFNλ increased PD-L1 and PD-L2, and IFNß or IFNλ treatment plus poly I:C further increased both expressions. Treatment with IC87114 or transfection with siRNA targeting PI3K p110δ enhanced poly I:C-induced gene and protein expression of PD-L2, whereas IC87114 suppressed poly I:C-induced PD-L1. IC87114 enhanced poly I:C-induced gene expression of IFNß, IFNλ, and IFN-regulated genes via increased TBK1 and IRF3 phosphorylation. Transfection with siIRF3 counteracted the enhancement of poly I:C-induced PD-L2 by IC87114, whereas IC87114 suppressed poly I:C-induced PD-L1 regardless of transfection with siNC or siIRF3. Similar effects of IC87114 on PD-L1 and PD-L2 expression were observed in human metapneumovirus-infected PBECs. We showed for the first time that type I and type III IFNs induced the expression of PD-L1 and PD-L2 in PBECs. Our findings suggest that during viral infections, inhibition of PI3Kδ differentially regulates PD-L1 and PD-L2 expression in bronchial epithelial cells.

Untangling radiocesium dynamics of forest-stream ecosystems: A review of Fukushima studies in the decade after the accident.

Forest-stream ecosystems are widespread and biodiverse terrestrial landscapes with physical and social connections to downstream human activities. After radiocesium is introduced into these ecosystems, various material flows cause its accumulation or dispersal. We review studies conducted in the decade after the Fukushima nuclear accident to clarify the mechanisms of radiocesium transfer within ecosystems and to downstream areas through biological, hydrological, and geomorphological processes. After its introduction, radiocesium is heavily deposited in the organic soil layer, leading to persistent circulation due to biological activities in soils. Some radiocesium in soils, litter, and organisms is transported to stream ecosystems, forming contamination spots in depositional habitats. While reservoir dams function as effective traps, radiocesium leaching from sediments is a continual phenomenon causing re-contamination downstream. Integration of data regarding radiocesium dynamics and contamination sites, as proposed here, is essential for contamination management in societies depending on nuclear power to address the climate crisis.

Airway compliance measurements in mouse models of respiratory diseases.

The quantification of airway compliance (Caw) is essential to the study of airway alterations in disease models. However, the required measurements of airway pressure and volume are difficult to acquire in mice. We hypothesized that the inflation limb of full-range pressure-volume (PV) curves could be used to quantify Caw, as it contains a segment where only the airway tree is distended. The study objective was to assess the feasibility of the approach by analysis of full-range PV curves previously collected in three mouse models: an elastase model of emphysema, a genetic model spontaneously developing emphysema (leukotriene C4 synthase knockout; LTC4S-KO), and a bleomycin model of lung fibrosis. Attempts to validate results included Caw change relative to respiratory system compliance (ΔCaw/ΔC), the minute work of breathing (mWOB), and the elastance at 20.5 Hz (Ers_20.5) from prior respiratory mechanics measurements in the same subjects. Caw was estimated at 3% of total compliance in healthy mice or 2.3 ± 1 µL/cmH2O (n = 17). The technique detected changes in models of respiratory obstructive and restrictive diseases relative to control mice as well as differences in the two emphysema models studied. The changes in Caw were consistent with those seen in ΔCaw/ΔC, mWOB, or Ers_20.5, with some variations according to the model, as well as with results reported in the literature in humans and mice. Direct Caw measurements in subjects as small as mice could prove useful to further characterize other respiratory disease models associated with airway remodeling or to assess treatment effects.

Incense smoke-induced oxidative stress disrupts tight junctions and bronchial epithelial barrier integrity and induces airway hyperresponsiveness in mouse lungs.

Recent clinical studies have suggested that inhalation of incense smoke (IS) may result in impaired lung function and asthma. However, there is little experimental evidence to link IS with airway hyperresponsiveness (AHR) and bronchial epithelial barrier function. Using mouse and cell culture models, we evaluated the effects of IS exposure on AHR, expression of multiple epithelial tight junction (TJ)- and adherens junction-associated mRNAs and proteins in the lungs, and the barrier function of bronchial epithelial cells assessed by transepithelial electronic resistance (TEER). Exposure of BALB/c mice to IS increased AHR and inflammatory macrophage recruitment to BALF; reduced claudin-1, -2, -3, -7, -10b, -12, -15, and -18, occludin, zonula occludens-1 [ZO-1], and E-cadherin mRNA expression; and caused discontinuity of claudin-2 and ZO-1 protein immunostaining in lung tissue. IS extract dose-dependently decreased TEER and increased reactive oxygen species production in bronchial epithelial cell cultures. Treatment with N-acetyl-L-cysteine, but not glucocorticosteroids or long-acting ß2-agonists, prevented the detrimental effects of IS. IS exposure can be problematic for respiratory health, as evidenced by AHR, increased recruitment of inflammatory macrophages and disruption of TJ proteins in the lung, and damage to epithelial barrier function. However, antioxidants may be useful for the treatment of IS-induced airway dysfunction.

Data on 137Cs concentration factor of freshwater fish and aquatic organisms in lake and river ecosystems.

This article provides the data which were analyzed in the research article "Different factors determine 137Cs concentration factors of freshwater fish and aquatic organisms in lake and river ecosystems" (Y. Ishii, S. S. Matsuzaki, S. Hayashi, 2019) [1]. Radionuclide accumulation in aquatic organism is defined in terms of the concentration factor (CF), which is calculated as the radionuclide concentration in the organism (Bq kg-1) divided by that in the surrounding water (Bq L-1). Quantification of the radionuclide CF allows estimation of environmental radionuclide transfer and the potential risks of consuming fish contaminated with the radionuclide. We calculated the 137Cs CF values for freshwater fish and aquatic organisms using the monitoring data of multiple sites in five rivers and three lakes of Fukushima in years 2013-2017 after the Fukushima Dai-ichi Nuclear Power Plant accident. The data also include the 137Cs activity concentration of the water and water chemistry data (pH, biochemical oxygen demand, chemical oxygen demand, dissolved oxygen, electric conductivity, salinity, total organic carbon, suspended solid concentration, turbidity) at each sampling location associated with each CF value.

Different factors determine 137Cs concentration factors of freshwater fish and aquatic organisms in lake and river ecosystems.

Determination of radionuclide concentration factor (CF) allows estimating the transfer of environmental radionuclides and potential risks of consuming fish contaminated with radionuclides. Although it is known that biotic and abiotic factors affect fish CF, only a few studies have examined whether these factors differ among ecosystems. We estimated radiocesium (137Cs) CF of 30 different fish species and other aquatic organisms by monitoring three lakes and five rivers in Fukushima, 2-4 y after the Fukushima Dai-ichi Nuclear Power Plant accident. The relative effects of biotic and abiotic factors on 137Cs CF in freshwater organisms were compared between river and lake ecosystems using generalized linear models. Our analysis demonstrated the following. (1) The factors critically affecting fish CF differed between rivers and lakes. The negative effects of suspended solid concentration (SS), total organic carbon (TOC), and salinity were significant for rivers, but not for lakes. Biomagnification of 137Cs in piscivore fish was significant only in the lakes. (2) Fish size significantly affected the CF in both rivers and lakes. Nevertheless, the correlation between 137Cs concentration and piscivore fish size was stronger in lakes than in rivers. (3) The SS, TOC, and salinity simultaneously influenced the CF at every trophic level. However, feeding habit was a stronger determinant of 137Cs bioaccumulation than water chemistry in organisms at higher trophic levels, such as aquatic insects, amphibians, and fish. Our findings indicate that 137Cs accumulation in aquatic organisms is ecosystem-dependent due to different environmental factors and food web structure.

Effects of cigarette smoke on barrier function and tight junction proteins in the bronchial epithelium: protective role of cathelicidin LL-37.

BACKGROUND: Airway epithelial barrier function is maintained by the formation of tight junctions (TJs) and adherens junctions (AJs). Inhalation of cigarette smoke causes airway epithelial barrier dysfunction and may contribute to the pathogenesis of chronic lung diseases such as asthma and chronic obstructive pulmonary disease (COPD). We assessed the effects of cigarette smoke on barrier function and expression of multiple TJ and AJ proteins in the bronchial epithelium. We also examined whether treatment with glucocorticosteroids (GCSs), long-acting ß2-agonists (LABAs), and human cathelicidin LL-37 can protect against cigarette smoke extract (CSE)-induced barrier dysfunction. METHODS: Calu-3 cells cultured at the air-liquid interface were pretreated with or without GCSs, LABAs, GCSs plus LABAs, or LL-37, and subsequently exposed to CSE. Barrier function was assessed by transepithelial electronic resistance (TEER) measurements. Gene and protein expression levels of TJ and AJ proteins were analyzed by quantitative PCR and western blotting, respectively. Immunofluorescence staining of TJ and AJ proteins was performed. RESULTS: CSE decreased TEER and increased permeability in a concentration-dependent manner. CSE suppressed gene expression of claudin-1, claudin-3, claudin-4, claudin-7, claudin-15, occludin, E-cadherin, junctional adhesion molecule-A (JAM-A) and zonula occludens-1 (ZO-1) within 12 h post-CSE exposure, while suppressed protein expression levels of occludin at 12 h. CSE-treated cells exhibited discontinuous or attenuated immunostaining for claudin-1, claudin-3, claudin-4, occludin, ZO-1, and E-cadherin compared with untreated cells. GCS treatment partially restored CSE-induced TEER reduction, while LABA treatment had no effect. GCS and LABA combination treatment had no additive effect on CSE-induced TEER reduction and gene suppression of TJ and AJ proteins. Human cathelicidin LL-37 counteracted CSE-induced TEER reduction and prevented disruption of occludin and ZO-1. LL-37 also attenuated CSE-induced decreases in gene and protein expression levels of occludin. CONCLUSIONS: CSE caused airway epithelial barrier dysfunction and simultaneously downregulated multiple TJ and AJ proteins. GCS and LABA combination treatment had no additive effect on CSE-induced TEER reduction. LL-37 counteracted CSE-induced TEER reduction and prevented disruption of occludin and ZO-1. Use of LL-37 to counteract airway epithelial barrier dysfunction may have significant benefits for respiratory diseases such as asthma and COPD.

Factors controlling dissolved 137Cs concentrations in east Japanese Rivers.

To investigate the main factors that control the dissolved radiocesium concentration in river water in the area affected by the Fukushima Daiichi nuclear power plant accident, the correlations between the dissolved 137Cs concentrations at 66 sites normalized to the average 137Cs inventories for the watersheds with the land use, soil components, topography, and water quality factors were assessed. We found that the topographic wetness index is significantly and positively correlated with the normalized dissolved 137Cs concentration. Similar positive correlations have been found for European rivers because wetland areas with boggy organic soils that weakly retain 137Cs are mainly found on plains. However, for small Japanese river watersheds, the building area ratio in the watershed strongly affected the dissolved 137Cs concentration. One reason for this would be because the high concentrations of solutes, such as K+ and dissolved organic carbon, discharged in urban areas would inhibit 137Cs absorption to soil particles. A multiple regression equation was constructed to predict the normalized dissolved 137Cs concentration with the topography, land use, soil component, and water quality data as explanatory variables. The best model had the building land use as the primary predictor. When comparing two multiple regression models in which the explanatory variables were limited to (1) the land use and soil composition and (2) the water quality, the water quality model underestimated the high normalized dissolve 137Cs concentration in urban areas. This poor reproducibility indicates that the dissolved 137Cs concentration value in urban areas cannot be solely explained by the solid-liquid distribution of 137Cs owing to the influence of the water quality, but some specific 137Cs sources in urban areas would control the dissolved 137Cs concentration.

Frequency-dependent airway hyperresponsiveness in a mouse model of emphysema and allergic inflammation.

Asthma and chronic obstructive pulmonary disease (COPD), chronic airway inflammatory diseases characterized by airflow limitation, have different etiologies and pathophysiologies. Asthma-COPD Overlap (ACO) has recently been used for patients with mixed asthma and COPD. The pathophysiological mechanisms of ACO have not been clearly understood due to the lack of an appropriate murine model. To investigate its pathophysiology, we examined a murine model by allergen challenge in surfactant protein-D (SP-D)-deficient mice that spontaneously developed pulmonary emphysema. SP-D-deficient mice were sensitized and challenged by ovalbumin (OVA). Lungs and bronchoalveolar lavage fluid (BALF) were collected for analysis, and static lung compliance and airway hyperresponsiveness (AHR) were measured 48 h after the last OVA challenge. In SP-D-deficient, naïve, or OVA-challenged mice, the mean linear intercept and static lung compliance were increased compared with wild-type (WT) mice. There was no significant difference in goblet cell hyperplasia and the gene expression of Mucin 5AC (MUC5AC) between SP-D-deficient and WT OVA-challenged mice. In SP-D-deficient OVA-challenged mice, airway hyperresponsiveness was significantly enhanced despite the lower eosinophil count and the concentration of interleukin (IL)-5 and IL-13 in BALF compared with WT OVA-challenged mice at 120 ventilations per minute. When mice were ventilated at a lower ventilation frequency of 100 ventilations per minute, elevated airway hyperresponsiveness in SP-D-deficient OVA-challenged mice was diminished. This model of emphysematous change with allergic airway inflammation raises the possibility that frequency-dependent airway hyperresponsiveness may be involved in the pathophysiology of ACO.

Radiocesium Transfer in Forest Insect Communities after the Fukushima Dai-ichi Nuclear Power Plant Accident.

To understand radiocesium transfer in the forest insect food web, we investigated the activity concentrations of radiocesium in forest insects in the Fukushima and Ibaraki Prefectures approximately 1.5-2.5 years after the Fukushima Dai-ichi Nuclear Power Plant. We analyzed 34 species of insects sampled from 4 orders and 4 feeding functional groups (herbivore, carnivore, omnivore, and detritivore) from three sites in each prefecture. 137Cs activity concentrations were lowest in herbivorous species and were especially high in detritivorous and omnivorous species that feed on forest litter and fungi. Radiocesium activity concentrations in any given species reflected the degree of contamination of that species' primary food sources since radiocesium activity concentrations were found to be the lowest in leaves and grass and the highest in litter, bark, and fungi. This study confirmed that litter and other highly contaminated forest components such as fungi, decaying wood, bryophytes, and lichens serve as sources of 137Cs transfer into the forest insect community.

Leukotriene B4 receptor type 2 protects against pneumolysin-dependent acute lung injury.

Although pneumococcal infection is a serious problem worldwide and has a high mortality rate, the molecular mechanisms underlying the lethality caused by pneumococcus remain elusive. Here, we show that BLT2, a G protein-coupled receptor for leukotriene B4 and 12(S)-hydroxyheptadecatrienoic acid (12-HHT), protects mice from lung injury caused by a pneumococcal toxin, pneumolysin (PLY). Intratracheal injection of PLY caused lethal acute lung injury (ALI) in BLT2-deficient mice, with evident vascular leakage and bronchoconstriction. Large amounts of cysteinyl leukotrienes (cysLTs), classically known as a slow reactive substance of anaphylaxis, were detected in PLY-treated lungs. PLY-dependent vascular leakage, bronchoconstriction, and death were markedly ameliorated by treatment with a CysLT1 receptor antagonist. Upon stimulation by PLY, mast cells produced cysLTs that activated CysLT1 expressed in vascular endothelial cells and bronchial smooth muscle cells, leading to lethal vascular leakage and bronchoconstriction. Treatment of mice with aspirin or loxoprofen inhibited the production of 12-HHT and increased the sensitivity toward PLY, which was also ameliorated by the CysLT1 antagonist. Thus, the present study identifies the molecular mechanism underlying PLY-dependent ALI and suggests the possible use of CysLT1 antagonists as a therapeutic tool to protect against ALI caused by pneumococcal infection.

Practical selection methods for rat and mouse round spermatids without DNA staining by flow cytometric cell sorting.

Round spermatid injection (ROSI) into unfertilized oocytes enables a male with a severe spermatogenesis disorder to have children. One limitation of the application of this technique in the clinic is the identification and isolation of round spermatids from testis tissue. Here we developed an efficient and simple method to isolate rodent haploid round spermatids using flow cytometric cell sorting, based on DNA content (stained with Hoechst 33342 or Dye Cycle Violet) or by cell diameter and granularity (forward and side scatter). ROSI was performed with round spermatids selected by flow cytometry, and we obtained healthy offspring from unstained cells. This non-invasive method could therefore be an effective option for breeding domestic animals and human male infertility treatment. Mol. Reprod. Dev. 83: 488-496, 2016. © 2016 Wiley Periodicals, Inc.

Prevalence of Airflow Limitation Defined by Pre- and Post-Bronchodilator Spirometry in a Community-Based Health Checkup: The Hisayama Study.

Spirometry in health checkup may contribute to early diagnosis of chronic obstructive pulmonary disease (COPD) and asthma. Although post-bronchodilator airflow limitation is essential for definite diagnosis of COPD and post-bronchodilator normalization of airflow is suggestive of asthma, this test has not been prevailed in health checkup. The objective of this study was to estimate the prevalence of airflow limitation defined by pre- and post-bronchodilator spirometry in health checkup. Post-bronchodilator spirometry was conducted for participants with airflow limitation in a town-wide health checkup for residents aged 40 years and older in Hisayama, a town in the western part of Japan. The prevalence of pre- and post-bronchodilator airway limitation defined by FEV1/FVC < 70% were estimated. A total of 2,232 participants underwent pre-bronchodilator spirometry. In males, the age of current smokers was significantly younger than those of never smokers and former smokers. In females, the ages of current- and former smokers were significantly younger than never smokers. The values of %FEV1 and %FVC in current smokers were significantly lower than those in former smokers and never smokers. Two hundred sixty nine subjects, 85% of total subjects with a pre-bronchodilator FEV1/FVC < 70%, completed post-bronchodilator spirometry. The prevalence of pre-bronchodilator airflow limitation was 14.6% in males and 13.7% in females, and the prevalence of post-bronchodilator airway limitation was 8.7% and 8.7%, respectively. Post-bronchodilator spirometry in health checkup would reduce the number of subjects with probable COPD to two-third. Recommendation for those examinees to take further evaluations may pave the way for early intervention.

Leukotriene B4 receptor type 2 (BLT2) enhances skin barrier function by regulating tight junction proteins.

GPCRs are involved in numerous physiologic functions and are important drug targets. Although the epithelial barrier is important for protection from invading pathogens, the correlation between GPCRs and epithelial barrier function remains unknown. Leukotriene B4 (LTB4) receptor type 2 (BLT2), mainly expressed in epithelial cells, is a GPCR for 12(S)-hydroxyheptadeca-5Z,8E,10E-trienoic acid (12-HHT) and LTB4. In our study, BLT2 localized at the lateral membrane in BLT2-overexpressing Madin-Darby canine kidney (MDCK) II cells and in the small intestine of BLT2-transgenic mice. BLT2-deficient mice exhibited higher transepidermal water loss and were more sensitive to epicutaneous sensitization. MDCK-BLT2 cells recovered transepithelial electrical resistance (TER) after a calcium switch faster than did MDCK-Mock cells, and 12-HHT stimulation accelerated TER recovery only in MDCK-BLT2 cells. Quantitative PCR and immunoblot analyses revealed that the 12-HHT/BLT2 axis up-regulated claudin-4 (CLDN4) expression in MDCK-BLT2 cells and human primary keratinocytes, and CLDN4 knockdown abolished 12-HHT-dependent TER recovery. Acceleration of TER recovery and induction of CLDN4 expression by 12-HHT stimulation were abolished by inhibition of Gαi protein or p38 MAPK. These results show that 12-HHT/BLT2 enhances epithelial barrier function by increasing CLDN4 expression via the Gαi protein-p38 MAPK pathway.