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COVID-19 causes physical and psychological impacts on health care workers (HCWs), especially when it occurs during an outbreak. As there are few reports on outcomes of HCWs infected with COVID-19 during a hospital outbreak, we investigated the physical and psychological impacts on HCWs infected with COVID-19 during an outbreak in our hospital.During the outbreak in our hospital, 231 people were infected with COVID-19 including patients, HCWs and their families. Among them, 83 HCWs were enrolled in this study. Current quality of life (QOL) was assessed with the EuroQol-visual analogue scales (EQ-VAS), and motivation to keep on working was evaluated by a 10-point analogue scale. Physiological recovery rates including return to work (RTW) period were also analyzed.One nurse quit work due to anxiety regarding re-infection with COVID-19. The median period to RTW from the diagnosis was 14.0 (12.0-17.0) days. Motivation to keep on working was slightly reduced, and the EQ-VAS was 75.0 (65.0-83.6). There were no significant differences in QOL and motivation between male and female HCWs, nurses and other HCWs, treatment and non-treatment group, and supplemental and non-supplemental oxygen group. The most frequent persistent symptoms at 1,3 and 6 months after infection were anosmia followed by fatigue.Although QOL and motivation to keep on working were slightly reduced, only one HCW quit work. No severe persistent symptoms were observed, and the RTW period was relatively short.

A 62-year-old woman was diagnosed with peritoneal dissemination of gastric cancer and was treated with anticancer drugs. Eleven months after the start of the treatment, follow-up computed tomography newly showed thickening of the bladder wall and left hydronephrosis even though the chemotherapy reduced peritoneal dissemination. Therefore, she was referred to our hospital for further evaluation. Cystoscopy and magnetic resonance imaging showed the tumor arising from the bladder neck to trigone. A few days later, she was admitted to our hospital because of bladder tamponade. Transurethral coagulation was carried out, and we resected part of the bladder tumor for pathological examination at the same time. As the pathological features of the bladder tumor were similar to those of the primary stomach cancer and peritoneal dissemination, the diagnosis of the bladder tumor was metastatic gastric adenocarcinoma. She died three months after visiting our hospital.

We conducted a multicenter, retrospective study to evaluate the efficacy and safety of combination nivolumab plus ipilimumab (NIVO+IPI) in 35 patients with advanced or metastatic renal cell carcinoma (mRCC). In this study, we focused on patients who received NIVO+IPI and were stratified into intermediate- or poor-risk disease according to the International Metastatic Renal Cell Carcinoma Database Consortium model at five institutions in Japan. The primary endpoint was overall survival (OS). Secondary endpoints were disease control rate (DCR), best overall response (BOR), objective response rate (ORR), and progression-free survival (PFS). In addition, we evaluated the role of inflammatory cell ratios, namely neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR), as predictive biomarkers in patients with mRCC. The median follow-up period was 1 year, and the 1-year OS rate was 95.8%. The ORR and DCR were 34.3% and 80.0%, respectively. According to BOR, four patients (11.4%) achieved complete response. According to NLR stratification, the 1-year PFS rates were 82.6% and 23.7% when the NLR was ≤4.6 and &gt, 4.6, respectively (p = 0.04). Based on PLR stratification, the 1-year PFS rates were 81.7% and 34.3% when the PLR was ≤188.1 and &gt, 188.1, respectively (p = 0.033). Although 71.4% of the patients experienced treatment-related adverse events (TRAEs) with NIVO+IPI, only four patients discontinued NIVO+IPI due to grade 3/4 TRAEs. Patients treated with NIVO+IPI as a first-line therapy for advanced or mRCC achieved relatively better oncological outcomes. Therefore, NIVO+IPI may have potential advantages and may lead to a treatment effect compared to those receiving targeted therapies. In addition, PLR &gt, 188.1 may be a useful predictive marker for mRCC patients who received NIVO+IPI.

The bioavailability of cadmium (Cd) in agricultural soils is a significant health concern due to the potential risk of human exposure via foods grown in Cd-contaminated fields. Biochar has been known to have a highly porous structure and high pH, as well as containing various functional groups; as such, it can immobilize heavy metals. Although it has found that biochar amendment in Cd-contaminated agricultural soils could be effective in reducing Cd bioavailability in previous studies, differences in plant Cd accumulation from Cd-contaminated soils amended with biochars produced from various types of biomass have not been fully discussed yet; we aimed to address this shortcoming in the present work. The soil investigated was an acid soil (pH 5.1) and had an elevated concentration of Cd (total Cd: 3.3 mg kg-DW−1). Six kinds of biochar were produced, i.e., from woodchips (Japanese cedar [CE] and Japanese cypress [CY]), moso bamboo (MB), rice husk (RH), poultry manure (PM), and wastewater sludge (WS), at a pyrolysis temperature of 600 °C. Biochars were incorporated into the Cd-contaminated soil at 3% (w/w) and pot experiments using Brassica rapa var. perviridis were conducted for 28 days in a growth chamber. The Cd concentrations in the above-ground portion of the plants were significantly decreased as a result of the incorporation of all biochars compared to the unamended soil, with reduction ratios following the order PM (78%) > > WS (31%) ≈ RH (29%) ≈ MB (28%) ≈ CY (26%) > CE (19%). Among all biochar-amended soils, soil pH and shoot biomass were highest for those amended with PM-derived biochar. These results suggest that in Cd-contaminated soils, PM-derived biochar may offer significant potential in reducing plant Cd accumulation due to the immobilization of soil Cd and an effect of dilution resulting from enhanced plant shoot biomass.

Objectives To assess the effect of cernitin pollen extract on serum prostate-specific antigen level prostate biopsy candidates, and to develop an ideal protocol to avoid an unnecessary biopsy procedure. Methods A total of 61 patients were administrated cernitin pollen extract tablets (two tablets t.i.d.) for 30 days, and then underwent a prostate biopsy with ≥12 systematic and targeted biopsy cores obtained. Serum prostate-specific antigen levels were examined before and after administration of the pollen extract, and the change in serum prostate-specific antigen and the rate of change were analyzed in relation to negative and positive biopsy results for cancer. Results The mean change in serum prostate-specific antigen and rate of change after administration of cernitin pollen extract in all patients were -0.6 ± 1.4 ng/mL and -7.6 ± 16.1%, respectively, which were significantly different from the baseline values (P = 0.0003 and P = 0.0005, respectively). When prostate-specific antigen change values and rates were compared between patients negative and positive for cancer, a significant difference between those groups was observed (P = 0.04 and P = 0.03, respectively). Conclusions The present study is the first to show that an ideal protocol using cernitin pollen extract has the potential to avoid an unnecessary prostate biopsy procedure in patients with elevated prostate-specific antigen, possibly caused by inflammation. Additional studies with greater numbers of participants are required to confirm our findings and develop an ideal protocol.

Physicochemical properties of biochar, which are used as a soil amendment material in agricultural fields, are different depending on biomass feedstock and pyrolysis processes. In this study, we evaluated the influence of feedstock type and pyrolysis temperature on the water-retention related properties of biochar. Wood-chips [cedar (CE) and cypress (CY)], moso bamboo (MB), rice husk (RH), sugarcane bagasse (SB), poultry manure (PM) and agricultural wastewater sludge (WS) were each pyrolysed at 400, 600 and 800 &deg, C with a retention time of two hours. Scanning electron microscopy micrographs (SEM), hydrophobicity indices, pore-size distribution measured by mercury-intrusion porosimetry, water-retention curves (WRCs) and plant-available water capacities (AWCs) of the biochars were measured to evaluate their potentials as soil-amendment materials for improving soils&rsquo, water-retention. As the pyrolysis temperature was increased, the hydrophobicity index decreased. On the other hand, pyrolysis temperature did not affect the distribution of micrometre-range pores, which are useful for plant-available water, of biochars. The AWCs of the biochars formed from CE, CY and SB were greater than those produced from other feedstocks, at 600 and 800 &deg, C. Therefore, we can suggest that the biochars derived from wood-chips (CE and CY) and SB have greater potential for enhancing soils&rsquo, water-retention.

Unlike urinary tract infection (UTI), asymptomatic bacteriuria (ABU) should not be treated, with some exceptions such as pregnant women and patients who will undergo traumatic urologic interventions. However, there has been no clinically available marker for their differential diagnosis. Exosomes or small extracellular vesicles carry proteins contained in cells from which they are derived, thus having the potential as a biomarker of several diseases. On the basis of the hypothesis that the molecular signature of exosomes in urine may differ between UTI and ABU patients, we examined if urinary exosomes could serve as a marker for their differential diagnosis. Exosomes were isolated by ultracentrifugation or affinity-based method from cell culture medium of monocytic THP-1 and uroepithelial SV-HUC-1 cells and human urine. Protein expression was examined by Western blot analysis, ELISA, and CLEIA. The results showed that the levels of intracellular signalling molecules Akt and ERK and transcription factor NF-κB increased in exosomes isolated from THP-1 and SV-HUC-1 cells cocultured with Escherichia coli and/or treated with lipopolysaccharide. In urinary exosomes of UTI patients, Akt significantly diminished, and an exosomal marker CD9 showed a trend to decrease after treatment with antimicrobial agents. More importantly, Akt and CD9 levels in urinary exosomes were higher in UTI patients than in ABU patients, which was also observed after correction by urine creatinine. Collectively, these results suggest that Akt and CD9 in urinary exosomes could be useful markers for differential diagnosis of UTI and ABU.

Biochar (BC), charcoal produced through the pyrolysis of biomass, is reported to adsorb dissolved nitrate-nitrogen (NO3-N). The NO3-N adsorption properties of BC differ depending on the feedstock and the pyrolysis conditions, and the influences have not been systematically clarified. Therefore, we evaluated the dependence of feedstock and pyrolysis temperature on the NO3-N adsorption properties of BC. Wood chips [Japanese cedar [Cryptomeria japonica] (CE) and Japanese cypress [Chamaecyparis obtusa] (CY)], moso bamboo [Phyllostachys edulis] chips (MB), rice [Oryza sativa] husks (RH), sugarcane [Saccharum officinarum] bagasse (SB), poultry manure (PM) and domestic wastewater sludge (WS) were air-dried and heated in a batch-type carbonization furnace at pyrolysis temperatures of 400, 600 and 800°C, with a hold time of 2 h. Among the BC produced from each feedstock, the one produced at 800°C had the greatest NO3-N adsorption. The NO3-N adsorption by BC produced from wood-based biomass at 800°C was sig...

Biochar (BC) is a promising soil amendment that can enhance water retention and plant-available water capacity while mitigating CO2 emissions. We investigated the effect of sugarcane bagasse–derived BC on the water retention properties of a calcaric clay soil amended with 3% (wt/wt) BC produced at three pyrolysis temperatures (400°C, 600°C, and 800°C). For BC pyrolyzed at 800°C (BC800), water retention curves of soil amended at 1%, 5%, and 10% (wt/wt) were also measured. Water retention curves were measured immediately after amending soil with BC (all types and rates) and after a 180-day incubation period for soils amended with 3% BC. The hydrophobicity of BC pyrolyzed at 400°C (B400) was the highest of the three temperatures tested, resulting in the lowest water retained in soil amended with BC400, but only for measurements done before incubation. During incubation, the hydrophobicity of B400 decreased as the aliphatic compounds became exhausted by oxidation of the BC surfaces. The available water capacity of the clay soil increased significantly by more than 60% when amended with BC at rates greater than of 3% wt/wt (P < 0.05).

To the Editor: In 2009, 2010, and 2013, Neisseria gonorrhoeae strains H041 (ceftriaxone MIC of 2 mg/L), F89 (ceftriaxone MIC of 1 mg/L), and A8806 (ceftriaxone MIC of 0.5 mg/L) were isolated from samples from patients in Japan (1), France (2) and Australia (3), respectively. In Japan, no other clinical N. gonorrhoeae strains with decreased susceptibility to ceftriaxone were reported until 2014, when clinical strain {"type":"entrez-nucleotide","attrs":{"text":"GU140106","term_id":"268370439","term_text":"GU140106"}}GU140106 (ceftriaxone MIC of 0.5 mg/L) was isolated from a man in in Nagoya, Japan. We report details of this case and sequencing results of the penA gene for the strain. The study was approved by the Institutional Review Board of the Graduate School of Medicine, Gifu University, Japan. N. gonorrhoeae strain {"type":"entrez-nucleotide","attrs":{"text":"GU140106","term_id":"268370439","term_text":"GU140106"}}GU140106 was isolated from a urethral swab sample from a man with acute urethritis. The man had received fellatio, without condom use, from a female sex worker in Nagoya in December 2013. He visited our clinic in January 2014 for urethral discharge. Culture of a urethral swab sample was positive for N. gonorrhoeae. We used the Cobas 4800 CT/NG Test (Roche Molecular Systems Inc., Pleasanton, CA, USA) to test a first-voided urine sample; results were positive for N. gonorrhoeae but negative for Chlamydia trachomatis. The infection was treated with a single-dose regimen of ceftriaxone (1 g) administered by intravenous drip infusion. Two weeks later, the man reported no symptoms, and his first-voided urine sample was negative for leukocytes. The test-of-cure for N. gonorrhoeae was not performed. The female sex worker could not be examined for the presence of N. gonorrhoeae strain {"type":"entrez-nucleotide","attrs":{"text":"GU140106","term_id":"268370439","term_text":"GU140106"}}GU140106 in her pharynx. The strain was confirmed to be a gonococcal species by testing with Gonochek-II (TCS Biosciences Ltd, Buckingham, UK), the HN-20 Rapid system identification test (Nissui, Tokyo, Japan), and the Aptima Combo 2 assay for CT/NG (Hologic, Inc., Bedford, MA, USA) and by 16S rRNA gene sequencing and porA pseudogene PCR (4). MICs of antimicrobial drugs for {"type":"entrez-nucleotide","attrs":{"text":"GU140106","term_id":"268370439","term_text":"GU140106"}}GU140106 were as follows, as determined by using the agar dilution method: 2.0 mg/L for penicillin G, 1.0 mg/L for tetracycline, 2.0 mg/L for cefixime, 0.5 mg/L for ceftriaxone, 8.0 mg/L for levofloxacin, 0.5 mg/L for azithromycin, and 32.0 mg/L spectinomycin. The strain was determined to be resistant to penicillin G, tetracycline, cefixime, ceftriaxone, and levofloxacin, according to criteria of the European Committee on Antimicrobial Susceptibility Testing (5) The penA gene of strain {"type":"entrez-nucleotide","attrs":{"text":"GU140106","term_id":"268370439","term_text":"GU140106"}}GU140106 was sequenced as previously described (6); results showed the presence of a novel mosaic penicillin-binding protein 2 (PBP2; GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"LC056026","term_id":"965690016","term_text":"LC056026"}}LC056026) (Figure). Multilocus sequence typing (MLST) and N. gonorrhoeae multiantigen sequence typing (NG-MAST) of {"type":"entrez-nucleotide","attrs":{"text":"GU140106","term_id":"268370439","term_text":"GU140106"}}GU140106 were performed as previously reported (8,9). MLST assigned strain {"type":"entrez-nucleotide","attrs":{"text":"GU140106","term_id":"268370439","term_text":"GU140106"}}GU140106 to sequence type 7363, the same as strains H041 and A8806 (1,2). NG-MAST assigned strain {"type":"entrez-nucleotide","attrs":{"text":"GU140106","term_id":"268370439","term_text":"GU140106"}}GU140106 to sequence type 6543. MLST and NG-MAST results for {"type":"entrez-nucleotide","attrs":{"text":"GU140106","term_id":"268370439","term_text":"GU140106"}}GU140106 differed from those for F89 (3). Figure Sequences of altered amino acids in penicillin-binding protein 2 (PBP2) of Neisseria gonorrhoeae strains with decreased susceptibility to oral cephalosporins and strains with resistance to ceftriaxone. Strain {"type":"entrez-nucleotide","attrs":{"text":"GU140106","term_id":"268370439","term_text":"GU140106"}} ... Since the naming of the mosaic PBP2 associated with decreased susceptibilities to oral cephalosporins as pattern X (6), various PBP2 mosaic structures have been discovered. Mosaic PBP2 structures are basically composed of fragments analogous to PBP2s in Neisseria species. Before strain H041 emerged, strains harboring mosaic PBP2s had been resistant to oral cephalosporins but susceptible to ceftriaxone. H041 (ceftriaxone MIC of 2 mg/L) had additional novel amino acid changes, including A311V, V316P, and T483S, in its mosaic PBP2. The presence of substitutions A311V, V316P, and T483S was reported to be responsible for resistance to ceftriaxone (7). Like strain H041, strains {"type":"entrez-nucleotide","attrs":{"text":"GU140106","term_id":"268370439","term_text":"GU140106"}}GU140106 and A8806 (ceftriaxone MICs of 0.5 mg/L) had substitutions A311V and T483S, but instead of substitution V316P, they had substitution V316T. In addition, {"type":"entrez-nucleotide","attrs":{"text":"GU140106","term_id":"268370439","term_text":"GU140106"}}GU140106 had several changes in positions 227–281 that were not present in other strains. These alterations might also contribute to the decreased susceptibility to ceftriaxone. On the basis of pharmacodynamic analyses (10), a 1-g dose of ceftriaxone (the recommended first-line treatment for gonorrhea in Japan) would be effective against genital gonorrhea caused by strains exhibiting decreased susceptibility to ceftriaxone (e.g., strains {"type":"entrez-nucleotide","attrs":{"text":"GU140106","term_id":"268370439","term_text":"GU140106"}}GU140106 and A8806). However, such strains could be resistant to lower-dose regimens, including 250-mg and 500-mg doses of ceftriaxone. This N. gonorrhoeae strain, {"type":"entrez-nucleotide","attrs":{"text":"GU140106","term_id":"268370439","term_text":"GU140106"}}GU140106, was isolated from the urethra of a man who received fellatio from a female sex worker; thus, the bacteria could have derived from her pharynx. N. gonorrhoeae strain H041 was previously isolated from the pharynx of a female sex worker (1). To prevent the emergence and spread of ceftriaxone-resistant N. gonorrhoeae, pharyngeal gonorrhea must be treated. It is uncertain whether a 1-g dose of ceftriaxone would be effective against pharyngeal gonorrhea caused by strains with decreased susceptibility to ceftriaxone, and this regimen might facilitate the selection of such strains from oral cephalosporin-resistant strains in the pharynx. The emergence of N. gonorrhoeae {"type":"entrez-nucleotide","attrs":{"text":"GU140106","term_id":"268370439","term_text":"GU140106"}}GU140106 in Japan suggests that new strategies (not just increased ceftriaxone doses), including combination treatment with ceftriaxone and another class of antimicrobial drugs and multiple dose regimens of ceftriaxone, might be required to treat pharyngeal gonorrhea.

Exosomes or microvesicles that are secreted from cells are considered to play important roles in tumor microenvironment. Carbonic anhydrase 9 (CA9), which is induced by hypoxia-inducible factor 1 (HIF1) in response to hypoxia, is overexpressed in many types of cancer including renal cell carcinoma (RCC). We examined the expression level of CA9 in several RCC cell lines and found that the basal level of CA9 was much higher in OSRC-2 cells than in Caki-1, KMRC-1 and 786-O cells. Consistent with the intracellular expression levels, CA9 was abundantly detected in exosomes isolated by ultracentrifugation from OSRC-2 cells. Density gradient centrifugation of OSRC-2 and 786-O exosomes confirmed the co-presence of CA9 with exosomal markers. Upon hypoxia and treatment with CoCl2, a hypoxia mimic agent, the CA9 level in exosomes was increased for all cell lines. In order to examine the effects of CA9 exosomes on angiogenesis, we generated stably transfected HEK293 cells expressing CA9. Immunocytochemical staining demonstrated the uptake of CA9 exosomes by human umbilical vein endothelial cells (HUVEC). In vitro angiogenesis assays using HUVEC revealed that CA9 exosomes promoted migration and tube formation. Lastly, MMP2 expression was increased by treatment with CA9 exosomes in HUVEC. Taken together, our results suggest the possibility that CA9 exosomes released from hypoxic RCC may enhance angiogenesis in microenvironment, thereby contributing to cancer progression.

Background Exosomes or extracellular vesicles have the potential as a diagnostic marker for various diseases including cancer. In order to identify novel exosomal markers for prostate cancer (PC), we performed proteomic analysis of exosomes isolated from PC cell lines and examined the usefulness of the marker in patients. Methods Exosomes isolated by differential centrifugation from the culture medium of androgen-dependent LNCaP prostate cancer cell line and its sublines of partially androgen-independent C4, androgen-independent C4–2 and bone metastatic C4–2B were subjected to iTRAQ-based proteomic analysis. Exosomes were also isolated by immunocapture and separated by size exclusion chromatography and density gradient centrifugation. Protein expression was determined by Western blot analysis. GGT activity was measured using a fluorescent probe, γ-glutamyl hydroxymethyl rhodamine green (gGlu-HMRG). Immunohistochemical analysis of tissues was performed using anti-GGT1 antibody. Results Among proteins upregulated in C4–2 and C4–2B cells than in LNCaP cells, we focused on gamma-glutamyltransferase 1 (GGT1), a cell-surface enzyme that regulates the catabolism of extracellular glutathione. The levels of both GGT1 large and small subunits were elevated in exosomes isolated from C4–2 and C4–2B cells by differential centrifugation and by immunocapture with anti-CD9 or -prostate-specific membrane antigen (PSMA) antibody. In cell lysates and exosomes, GGT1 expression correlated with GGT activity. Size exclusion chromatography of human serum demonstrated the presence of GGT activity and GGT1 subunits in fractions positive for CD9. Density gradient centrifugation revealed the co-presence of GGT1 subunits with CD9 in exosomes isolated by differential centrifugation from human serum. Since GGT activity correlated with GGT1 expression in serum exosomes isolated by differential centrifugation, we measured serum exosomal GGT activity in patients. Unexpectedly, we found that serum exosomal GGT activity was significantly higher in PC patients than in benign prostatic hyperplasia (BPH) patients. In support of this finding, immunohistochemical analysis showed increased GGT1 expression in PC tissues compared with BPH tissues. Conclusions Our results suggest that serum exosomal GGT activity could be a useful biomarker for PC. Electronic supplementary material The online version of this article (doi:10.1186/s12885-017-3301-x) contains supplementary material, which is available to authorized users.

In this article we have assessed the expected impacts of future climate changes on rainfall erosivity, a rainfall factor associated with soil erosion, of farmlands in Japan. The assessment compared values of R -factors for two future periods with values for a near past period. The R -factor is defined as the mean annual sum of individual storm erosion index values EI 30 , where E is the total energy for a storm and I 30 is the maximum 30-min intensity. The R -values at 1036 meteorological stations for three periods (1981–2000, 2031–2050 and 2081–2100) were calculated with a model that estimates monthly EI 30 values based on daily precipitation data. Subsequently, R -values for farmland areas were obtained. The daily precipitation data were provided from the regional climate model RCM20, which simulates climate change based on the SRES A2 scenario. The model to estimate monthly EI 30 values performed well with a coefficient of determination in excess of 0.5 for 97% of the 1036 stations when parameters of the model were optimized. Therefore, the rainfall erosivity model can be used to accurately estimate the R -factors. Calculations of relative changes in R -values for the two future periods compared to the near past period indicated that rainfall erosivity will increase in most farmland areas due to climate change. The R -values of farmland areas for the two future periods are expected to increase an average of more than 20% when compared to the near past. Additionally, changes in rainfall erosivity during each of the future periods are expected to vary with different seasonal and spatial patterns. The changes in erosivity appear to be attributed mainly to changes in precipitation intensity as well as amount of precipitation. Therefore, these results suggest the scenario of climate changes occurring in the future up to 2100 will increase soil erosion of farmlands in Japan at an average rate of increase greater than 20% based on just the effects of the rainfall erosivity factor.

A 26-year-old woman with gross hematuria was seen in a previous hospital. Magnetic resonance imaging (MRI) showed a tumor at the dome of the urinary bladder with invasion outside of the bladder wall. The patient underwent transurethral resection of the bladder tumor (TUR-BT). From the result of the pathological examination, the tumor was suggested to be carcinosarcoma of the bladder. The patient was then referred to our hospital for treatment. We performed radical cystectomy and ileal conduit diversion. Pathological examination of the excised specimen revealed an inflammatory myofibroblastic tumor as the basis for immunostaining of anaplastic lymphoma kinase (ALK).

Summary The incorporation of biochar (BC) into agricultural soil changes the soil's physical properties, which leads to changes in the soil's hydraulic properties, such as water retention and permeability, and alters the soil moisture environment in agricultural fields. To elucidate the effects of the incorporation of BC on the soil moisture environment, measurements of the soil water in biochar-amended agricultural fields are needed. Time domain reflectometry (TDR) is a widely used and established technique for the continuous measurement of the soil water content (SWC) in agricultural fields. However, TDR measurements are affected by the conductivity of soils. Biochar formed at higher pyrolysis temperatures is known to be very conductive. Therefore, we investigated the influence of the incorporation of BC on TDR-based SWC measurements. We examined calcaric dark red soil and the BC produced by pyrolysis of sugarcane bagasse at 400, 600 and 800°C. The apparent relative permittivity (ea) of the BC (800°C)-amended soil was greater than that of the non-amended soil at a given water content, whereas the ea values of the soils amended with the BC (400°C) and BC (600°C) were the same as that of the non-amended soil at a given water content. We concluded that when a calibration curve obtained from a non-amended soil is used, TDR-based measurements tend to over-estimate the SWC containing the BC formed at higher pyrolysis temperatures because of conductive and dielectric losses. Therefore, the use of the real component () of the soil's complex relative permittivity instead of ea is effective when making TDR-based water content measurements of soils that contain BC formed at higher pyrolysis temperatures.

With the increase in the production of bioethanol, it has become urgent to establish a sustainable system for recycling vinasse that includes its application to land. The purpose of this study was to determine the impact of vinasse application on the hydraulic properties of the soil. An incubation experiment was conducted to investigate the biological effects. Temporal changes in both saturated and unsaturated hydraulic conductivities [Kₛ and K(ψ), respectively] after applying vinasse were measured by the falling-head method and the steady-state flux control method, respectively. After vinasse application, the number of bacteria increased by one order of magnitude, whereas the number of fungi increased by two orders of magnitude compared with a test with deionized water. A remarkable reduction in Kₛ was observed within the fourth day after vinasse was applied instead of deionized water. The reduction in Kₛ was due to both biological and physical clogging. We attempted to determine what factors contributed to these effects on the Kₛ reduction and found that 73 to 92% of the Kₛ reduction was caused by physical clogging due to suspended solids. The temporal reduction in K(ψ) was within one order of magnitude; however, the values of K(ψ) dropped by two orders of magnitude for the same matric potential (ψ) value at the end of each column experiment. Moreover, 91% of the increase in the total amount of organic matter came from vinasse. From the experimental results, we concluded that the reductions in both Kₛ and K(ψ) occurred during vinasse application and the main reason for the reductions was the physical clogging due to suspended solids.

Advanced bladder cancer is treated mainly with gemcitabine and cisplatin, but most patients eventually become resistance. Androgen receptor (AR) signaling has been implicated in bladder cancer as well as other types of cancer including prostate cancer. In this study, we investigated the expression and role of AR in gemcitabine-resistant bladder cancer cells and also the potential of enzalutamide, an AR inhibitor, as a therapeutic for the chemoresistance. First of all, we established gemcitabine-resistant T24 cells (T24GR) from T24 bladder cancer cells and performed gene expression profiling. Microarray analysis revealed upregulation of AR expression in T24GR cells compared with T24 cells. AR mRNA and protein expression was confirmed to be increased in T24GR cells, respectively, by quantitative RT-PCR and western blot analysis, which was associated with more potent AR transcriptional activity as measured by luciferase reporter assay. The copy number of AR gene in T24GR cells determined by PCR was twice as many as that of T24 cells. AR silencing by siRNA transfection resulted in inhibition of proliferation of T24GR cells. Cell culture in charcoal-stripped serum and treatment with enzalutamide inhibited growth of T24GR cells, which was accompanied by cell cycle arrest. AR transcriptional activity was found to be reduced in T24GR cells by enzalutamide treatment. Lastly, enzalutamide also inhibited cell proliferation of HTB5 bladder cancer cells that express AR and possess intrinsic resistance to gemcitabine. Our results suggest that enzalutamide may have the potential to treat patients with advanced gemcitabine-resistant bladder cancer with increased AR expression.

Advanced renal cell carcinoma is treated with mammalian target of rapamycin (mTOR) inhibitors or tyrosine kinase inhibitors (TKIs). The effects of these drugs are, however, limited and novel treatment strategies are required. Clear-cell type renal cell carcinoma (ccRCC) is chemo-resistant, in part, due to expression of multidrug resistance proteins such as p-glycoprotein. Cabazitaxel, a tubulin-binding taxane drug used for castration-resistant prostate cancer, has less affinity for p-glycoprotein compared to docetaxel. In the current study, the effects of docetaxel and cabazitaxel on ccRCC cells were investigated.The expression of p-glycoprotein was evaluated in the ccRCC cell lines, Caki-1, KMRC-1 and OS-RC-2 by western blotting. Cells were treated with cabazitaxel or docetaxel, and growth kinetics and tubulin polymerization were determined by the WST-1 assay and cell-based tubulin polymerization assay, respectively. Intracellular drug concentrations were measured by chromatography. AKT activation after treatment was examined by western blotting.All ccRCC cell lines expressed p-glycoprotein. Cabazitaxel inhibited cell growth and induced tubulin polymerization more potently than docetaxel. The intracellular concentration of cabazitaxel was much higher than docetaxel in all cell lines. Both docetaxel and cabazitaxel inhibit AKT phosphorylation at 5 min among three cells.Cabazitaxel inhibits growth of ccRCC cells expressing p-glycoprotein and could thus be possibly used for advanced ccRCC patients in combination with targeted-therapy enhancing their effects.

Dielectric properties of soils are widely used to estimate their water content. Andisols are unique soils in terms of aggregate structure and show dielectric properties different from other mineral soils. To understand the relationship between the dielectric properties and soil physical properties, multi-phase dielectric mixing models are often used. However, solid phase permittivity (e s ) for Andisols, which is required for calculating the model output, has not been directly measured yet. Therefore, the objectives for this study were to measure e s for Andisols in Japan. In addition, the measured e s values were compared with those calculated from the traditional method, which applies two-phase mixing models to estimate e s values based on effective permittivities of repacked soil samples. The applicability of the traditional method to estimate such values for Andisols is also discussed. The effects of bound water and aggregate structure on measured e s values were evaluated prior to their measurements. We found that the aggregate structural effects were negligible. However, the amount of bound water caused overestimates of e s . Four Andisols from the A and B horizons of soils in Memuro Hokkaido (northern part of Japan), from the A horizon of a soil in Tsukuba Ibaraki (central part of Japan), and from the A horizon of a soil in Koshi Kumamoto (southern part of Japan) were used in this study. The e s values obtained fall between 5.6 and 6.1, and deviated from the estimated e s, values derived from the traditional method. Therefore, the traditional method is probably unsuitable to estimate e s values for Andisols.

Reddish sediment runoff from agricultural fields results in coastal environmental problems in Okinawa, Japan. Recent studies have demonstrated the effectiveness of strips of centipede grass (Eremochloa ophiuroides (Munro) Hack.), a perennial turf grass, in reducing the sediment loads from farmlands. However, sufficient information has not been provided to determine the appropriate strip specifications in the grass strip design. This study evaluated centipede grass strips for reduction of reddish sediment runoff from farmlands in Okinawa, Japan. A numerical model simulating the reddish sediment transport in the grass strip was constructed to determine the sediment removal efficiency of the strip. The model was verified using data obtained from field plot experiments with the grass strips under natural conditions. The sensitivity analysis of the model showed that the length of the grass strip (i.e. the dimension of the strip in the direction of flow) and unit inflow discharge have a great effect on sediment removal efficiency. The sediment removal efficiency obtained from the model simulation increased with the length of the strip and the increment of the efficiency decreased with the length of the strip. Therefore, these results indicate that the effective and efficient length of a centipede grass strip is 3 m for the reduction of reddish sediment loads under typical farmland conditions in Okinawa.

Objectives Docetaxel is used as the first-line chemotherapy for castration-resistant prostate cancer (CRPC), but docetaxel resistance occurs in part owing to induction of P-glycoprotein (P-gp) encoded by multidrug resistance protein 1 (MDR1) gene. A recently developed taxane—cabazitaxel—has poor affinity for P-gp and is thereby effective in docetaxel-resistant CRPC. It has been recently demonstrated that exosomes in the body fluids could serve as a diagnostic marker because they contain proteins and RNAs specific to the cells from which they are derived. In this study, we aimed to investigate if P-gp in blood exosomes could be a marker to diagnose docetaxel resistance and select a taxoid for patients with CRPC. Methods and materials Exosomes were isolated by differential centrifugation from docetaxel-resistant prostate cancer (PC-3) cells (PC-3R) and their parental PC-3 cells and from the serum of patients. Silencing of P-gp was performed by small interfering RNA transfection. Protein expression was examined by Western blot analysis. Viability of cells treated with docetaxel or cabazitaxel was determined by water soluble tetrazolium salt (WST) assay. Results The level of P-gp was higher in exosomes as well as cell lysates from PC-3R cells than in those from PC-3 cells. Cabazitaxel effectively killed PC-3R cells, and MDR1 knockdown improved the sensitivity of PC-3R cells to docetaxel but not to cabazitaxel. The P-gp level in blood exosomes was relatively higher in clinically docetaxel-resistant patients than in therapy-naive patients. Conclusions Our results suggest that detection of P-gp in blood exosomes, which is involved in resistance to docetaxel but not to cabazitaxel, could be useful to diagnose docetaxel resistance and select an appropriate taxoid for patients with CRPC—docetaxel or cabazitaxel.

Treatment with taxanes for castration-resistant prostate cancer often leads to the development of resistance. It has been recently demonstrated that exosomes present in the body fluids contain proteins and RNAs in the cells from which they are derived and could serve as a diagnostic marker for various diseases. In the present study, we aimed to identify proteins contained in exosomes that could be markers for progression and taxane-resistance of prostate cancer. Exosomes were isolated by differential centrifugation from the culture medium of taxane-resistant human prostate cancer PC-3 cells (PC-3R) and their parental PC-3 cells. Isolated exosomes were subjected to iTRAQ-based quantitative proteomic analysis. Exosomes were also isolated from the culture medium by using anti-CD9 antibody-conjugated magnetic beads. Protein expression was knocked down by siRNA transfection followed by analysis of the silencing effects. Proteomic analysis showed that integrin β4 (ITGB4) and vinculin (VCL) were upregulated in exosomes derived from PC-3R cells compared to PC-3 cells. The elevation of ITGB4 and VCL was confirmed in exosomes captured by anti-CD9 antibody from the culture medium of PC-3R cells. Silencing of ITGB4 and VCL expression did not affect proliferation and taxane-resistance of PC-3R cells, but ITGB4 knockdown attenuated both cell migration and invasion and VCL knockdown reduced invasion. Our results suggest that ITGB4 and VCL in exosomes could be useful markers for progression of prostate cancer associated with taxane-resistance, providing the basis for development of an exosome-based diagnostic system.

A 72-year-old Japanese man was referred to a hospital because of urinary retention. Digital rectal examination revealed a stony, hard nodule in the prostate. A high level of serum PSA was not detected. Prostatic biopsy was performed, and pathological examination indicated adenocarcinoma of the prostate. He was referred to our hospital for treatment. Imaging examinations revealed no metastases (T4N0M0), so we re-evaluated the biopsy specimens. Immunohistochemical examination revealed prostatic small cell carcinoma. His levels of neuron-specific enolase (NSE) and pro-gastrin-releasing peptide (Pro-GRP) were high. We treated him with combination chemotherapy comprising irinotecan and cisplatin, and the treatment was effective. After four courses of the chemotherapy, levels of NSE and Pro-GRP had decreased, and the prostatic mass had decreased in size. Needle rebiopsy of the prostate demonstrated no evidence of malignancy. Adjuvant external beam radiation therapy was also performed. The patient iss till alive at 18 month after diagnosis with no evidence of relapse or metastasis of the disease.

Purposes of the study: Exosome has been demonstrated to be a useful non-invasive biomarker for several cancers including prostate cancer. For better understanding cancer-derived exosome, their proteomic analysis is necessary. The aim of this study is to establish the method for isolation and analysis of prostate cancer-related exosomes. Experimental procedures: Total exosomes were isolated from the conditioned media of LNCaP cells by ultracentrifugation. Then, exosomes positive for CD9 or prostate specific membrane antigen (PSMA) were isolated by magnetic beads conjugated with anti-CD9 or -PSMA antibody, respectively. Isolated exosomes were subjected to proteomic analysis. The PSMA positive fraction was also collected by anti-PSMA beads from diluted serum of 3 prostate cancer patients and analyzed by proteomics. Results: A total of 126, 139 and 13 proteins were detected in the LNCaP exosome fractions that were isolated by ultracentrifugation, anti-CD9 beads and anti-PSMA beads, respectively. Out of 126 proteins identified in ultracentrifuge isolated exosome, 56 and 6 protein were found in exosomes that were isolated by anti-CD9 beads and anti-PSMA beads, respectively. Seven proteins were commonly detected in exosomes that were isolated by both beads. Fifty, 54 and 48 proteins were identified in the PSMA positive fraction from serum of 3 prostate cancer patients (P1: localized prostate cancer, P2: advanced prostate cancer and P3: castration resistant prostate cancer). Thirty-three proteins were detected in one patient, 22 in two patients and 25 in three patients. Most proteins detected in all patients were serum- or immunoglobulin-related proteins. The numbers of proteins that were detected only in P2 and P3 were 16 and 8, respectively. Six proteins were found in both P2 and P3 patients. Conclusion: In the present study, we analyzed prostate cancer-related exosomes that were isolated by immunoaffinity-based method. Although most proteins were serum- or immunoglobulin-related proteins, proteins that had been reported as a cancer marker were also detected, which could be candidates for exosomal marker of prostate cancer. Citation Format: Kosuke Mizutani, Kyojiro Kawakami, Yasunori Fujita, Kengo Horie, Koji Kameyama, Masafumi Ito, Takashi Deguchi. Proteomic analysis of prostate cancer-related exosomes isolated by anti-PSMA antibody beads [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5697. doi:10.1158/1538-7445.AM2017-5697

Exosomes have been demonstrated to be useful non-invasive biomarkers for several cancers including prostate cancer. Since normal cells also secrete exosomes, isolation of cancer-derived exosomes from blood is a prerequisite for their better understanding. The aim of this study is to establish the method for isolation of prostate cancer-related exosomes from blood.Exosomes were collected from prostate cancer LNCaP and PC-3 cell lines by ultracentrifugation and by using magnetic beads conjugated with anti-CD9 antibody and anti-prostate-specific membrane antigen (PSMA) antibody. Prostate cancer-related exosomes were also isolated from the plasma of prostate cancer patients by anti-PSMA beads. Isolated exosomes were analyzed by western blotting.Exosomes were isolated from LNCaP cells by ultracentrifugation, contained PSMA and androgen receptor (AR). AR was also detected in exosomes isolated from LNCaP cells by anti-PSMA and anti-CD9 beads, showing that AR is present in prostate cancer-related exosomes. The amount of CD9 in isolated exosomes was much higher in advanced and chemo-resistant prostate cancer patients than in prostate cancer patients without metastasis and healthy volunteers, indicating that patients with aggressive prostate cancer exhibit higher levels of prostate cancer-related exosomes in blood.The immunoaffinity-based method we developed is capable of isolating prostate cancer-related exosomes from blood, the use of which will enhance investigation processes on exosomes in prostate cancer.

Application of biochar has been suggested to improve water- and fertilizer-retaining capacity of agricultural soil. The objective of this study was to evaluate the effects of bagasse charcoal (sugarcane [ L.] bagasse-derived biochar) on nitrate (NO) leaching from Shimajiri Maji soil, which has low water- and fertilizer-retaining capacity. The nitrate adsorption properties of bagasse charcoal formed at five pyrolysis temperatures (400-800° C) were investigated to select the most suitable bagasse charcoal for NO adsorption. Nitrate was able to adsorb onto the bagasse charcoal formed at pyrolysis temperatures of 700 to 800° C. Nitrate adsorption by bagasse charcoal (formed at 800° C) that passed through a 2-mm sieve was in a state of nonequilibrium even at 20 h after the addition of 20 mg N L KNO solution. Measurements suggested that the saturated and unsaturated hydraulic conductivity of bagasse charcoal (800° C)-amended soils are affected by changes in soil tortuosity and porosity and the presence of meso- and micropores in the bagasse charcoal, which did not contribute to soil water transfer. In NO leaching studies using bagasse charcoal (800° C)-amended soils with different charcoal contents (0-10% [w/w]), the maximum concentration of NO in effluents from bagasse charcoal-amended soil columns was approximately 5% less than that from a nonamended soil column because of NO adsorption by bagasse charcoal (800° C). We conclude that application of bagasse charcoal (800°C) to the soil will increase the residence time of NO in the root zone of crops and provide greater opportunity for crops to absorb NO.

A 41-year-old woman with an incidental tumor of the urinary bladder was referred to our hospital. Computed tomography and magnetic resonance imaging showed a tumor in the urinary bladder wall with expansive growth. Under the suspicion of leiomyoma, we performed transurethral resection of the tumor. Pathological examination of tumor specimens revealed patternless arrangements of spindle cells. Immunochemical analysis revealed tumor cells positive for CD34 and bcl-2. The final diagnosis was a solitary fibrous tumor.