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Introduction: Antidiabetic drug metformin exerts various antitumor effects on different cancers. Esophageal squamous cell carcinoma (ESCC) is an intractable digestive organ cancer and new treatment strategy is required. In this study, we performed a comprehensive gene expression analysis of ESCC cell lines treated with metformin, which provided helpful information on the antitumor effects of metformin in ESCC. Next, we selected a promising gene among them and examined its effects on ESCC properties. Methods: We examined metformin-induced mRNA expression changes in two human ESCC cell lines by performing next-generation sequencing (NGS) and pathway analysis. Heat shock protein family A (Hsp70) member 6 (HSPA6) expression in surgical specimens obtained from 83 ESCC patients who underwent curative operations was evaluated immunohistochemically and analyzed. Results: Metformin upregulated mRNA expression of the many genes, including HSPA6, a cancer immune-related gene, and inhibited mRNA expression of the other many genes. Pathway analysis indicated major canonical pathways and upstream regulators related to metformin. The result indicated HSPA6 as a promising biomarker. HSPA6 expression correlated with disease-free survival (DFS) of the patients with all stage ESCC (p = 0.021), especially with stage I/II ESCC (p < 0.001). With stage III, low HSPA6 expression was not associated with poor DFS (p = 0.918). Multivariate analysis indicated that independent low HSPA6 expression was an independent poor prognostic factor of stage I/II ESCC (p < 0.001). However, HSPA6 expression did not correlate with the clinicopathological characteristics, including age, sex, tumor depth, lymph node metastasis, tumor stage, and tumor markers of the patients with stage I/II ESCC. Conclusions: This NGS analysis detected prospective candidate genes, including HSPA6. Our results indicate that HSPA6 is a promising biomarker of the recurrence risk of stage I/II ESCC. Further studies on HSPA6 would lead to better treatment.

This study indicated the novel impaired mechanism of ribosomal RNA (rRNA) transcription that causes cancer development. Human transcription factor IIH (TFIIH) is an essential factor with a seven-subunit core, including the P62 subunit, and is pivotal for DNA repair and transcription. Three RNA polymerases (RNAPs), RNAPI, II and III, possess a common RPB6 subunit that interacts with P62 of TFIIH. The FUBP1-interacting repressor (FIR) is a transcriptional suppressor of c-myc via suppressing DNA helicase activity of TFIIH. This study showed that FIR contains a highly conserved acidic string region, like the region of the RPB6 subunit of RNAPI. Knockdown of FIR by siRNA suppressed P62 expression in vivo in cancer cells. The isothermal titration calorimetry (ITC) measurements revealed the molecular interaction between the authentic FIR rather than FIRΔexon2, a dominant-negative splicing form of FIR that lacks the transcriptional repression domain exon2, and P62. Furthermore, knockdown of FIR and FIRΔexon2 by siRNA led to dynamic changes in ribosomal RNA (rRNA) expression assessed by RNA-seq analysis. A qRT-PCR analysis confirmed that FIR and FIRΔexon2 significantly changed the expression of numerous rRNAs on specific chromosomes. Moreover, small molecular chemical compounds against FIR and FIRΔexon2 were screened among 23,275 chemicals by Natural Product Depository (NPDepo) Array at RIKEN (Japan). Among them, BK697 that potentially binds FIR and FIRΔexon2 significantly inhibited tumor cell growth with the suppression of ribosomal proteins, FIR, and FIRΔexon2 expression. In summary, FIR and FIRΔexon2 provide a novel insight into cell proliferation by stimulating dynamic ribosome biosynthesis and a hint of therapeutic strategy.

Background Acute ischemic stroke (AIS) is a serious cause of mortality and disability. AIS is a serious cause of mortality and disability. Early diagnosis of atherosclerosis, which is the major cause of AIS, allows therapeutic intervention before the onset, leading to prevention of AIS. Methods Serological identification by cDNA expression cDNA libraries and the protein array method were used for the screening of antigens recognized by serum IgG antibodies in patients with atherosclerosis. Recombinant proteins or synthetic peptides derived from candidate antigens were used as antigens to compare serum IgG levels between healthy donors (HDs) and patients with atherosclerosis-related disease using the amplified luminescent proximity homogeneous assay-linked immunosorbent assay. Results The first screening using the protein array method identified death-inducer obliterator 1 (DIDO1), forkhead box J2 (FOXJ2), and cleavage and polyadenylation specificity factor (CPSF2) as the target antigens of serum IgG antibodies in patients with AIS. Then, we prepared various antigens including glutathione S-transferase-fused DIDO1 protein as well as peptides of the amino acids 297–311 of DIDO1, 426–440 of FOXJ2, and 607–621 of CPSF2 to examine serum antibody levels. Compared with HDs, a significant increase in antibody levels of the DIDO1 protein and peptide in patients with AIS, transient ischemic attack (TIA), and chronic kidney disease (CKD) but not in those with acute myocardial infarction and diabetes mellitus (DM). Serum anti-FOXJ2 antibody levels were elevated in most patients with atherosclerosis-related diseases, whereas serum anti-CPSF2 antibody levels were associated with AIS, TIA, and DM. Receiver operating characteristic curves showed that serum DIDO1 antibody levels were highly associated with CKD, and correlation analysis revealed that serum anti-FOXJ2 antibody levels were associated with hypertension. A prospective case–control study on ischemic stroke verified that the serum antibody levels of the DIDO1 protein and DIDO1, FOXJ2, and CPSF2 peptides showed significantly higher odds ratios with a risk of AIS in patients with the highest quartile than in those with the lowest quartile, indicating that these antibody markers are useful as risk factors for AIS. Conclusions Serum antibody levels of DIDO1, FOXJ2, and CPSF2 are useful in predicting the onset of atherosclerosis-related AIS caused by kidney failure, hypertension, and DM, respectively.

There is no clinically available biomarker for efficiently indicating the overall survival or therapy response of gastric cancer (GC). The autoantibodies (Abs) in the sera of anti‐far‐upstream element‐binding protein‐interacting repressor‐lacking exon2 (FIRΔexon2), anti‐sorting nexin 15, and anti‐spermatogenesis and oogenesis–specific basic helix–loop–helix 1 were markedly higher in GC patients than in healthy donors (HDs). These Abs were identified by large‐scale serological identification of antigens by recombinant cDNA expression cloning screenings and their expression levels were evaluated by amplified luminescence proximity homogeneous assay. In particular, compared with age‐matched HDs, the level of anti‐FIRΔexon2 Abs in GC patients was significantly higher (P, Anti‐FIRΔexon2 Ab is a predictive or overall survival marker for patients with gastric cancer.

Some cancers are related to atherosclerotic diseases; therefore, these two types of disease may share some antibody biomarkers in common. To investigate this, a first screening of sera was performed from patients with esophageal squamous cell carcinoma (ESCC) or acute ischemic stroke (AIS) for serological identification of antigens using recombinant cDNA expression cloning (SEREX). The amplified luminescent proximity homogeneous assay‐linked immunosorbent assay (AlphaLISA) method, which incorporates glutathione donor beads and anti‐human IgG acceptor beads, was used to evaluate serum antibody levels. SEREX screening identified low‐density lipoprotein receptor–related protein–associated protein 1 (LRPAP1) as a target antigen of serum IgG antibodies in the sera of patients with ESCC or AIS. Antigens, including recombinant glutathione S‐transferase–fused LRPAP1 protein, were prepared to examine serum antibody levels. AlphaLISA revealed significantly higher antibody levels against the LRPAP1 protein in patients with solid cancers such as ESCC and colorectal carcinoma and some atherosclerosis‐related diseases such as AIS and diabetes mellitus compared with healthy donors. Correlation analysis revealed that the elevated serum antibody levels against LRPAP1 were associated with smoking, a well‐known risk factor for both cancer and atherosclerosis. Serum LRPAP1 antibody is therefore a common marker for the early diagnosis of some cancers and atherosclerotic diseases and may reflect diseases caused by habitual smoking., Some cancers and arteriosclerosis may share some antibody biomarkers in common. To investigate this, a first screening of sera was performed from patients with esophageal squamous cell carcinoma for SEREX. Serum LRPAP1 antibody is a useful marker for the early diagnosis of some cancers and atherosclerotic diseases.

Black-odorous urban water bodies and sediments pose a serious environmental problem. In this study, we conducted microcosm batch experiments to investigate the effect of remediation reagents (magnesium hydroxide and calcium nitrate) on native bacterial communities and their ecological functions in the black-odorous sediment of urban water. The dominant phyla (Proteobacteria, Actinobacteria, Chloroflexi, and Planctomycetes) and classes (Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria, Actinobacteria, Anaerolineae, and Planctomycetia) were determined under calcium nitrate and magnesium hydroxide treatments. Functional groups related to aerobic metabolism, including aerobic chemoheterotrophy, dark sulfide oxidation, and correlated dominant genera (Thiobacillus, Lysobacter, Gp16, and Gaiella) became more abundant under calcium nitrate treatment, whereas functional genes potentially involved in dissimilatory sulfate reduction became less abundant. The relative abundance of chloroplasts, fermentation, and correlated genera (Desulfomonile and unclassified Cyanobacteria) decreased under magnesium hydroxide treatment. Overall, these results indicated that calcium nitrate addition improved hypoxia-related reducing conditions in the sediment and promoted aerobic chemoheterotrophy.

Recently, Wadi flash floods (WFFs) have happened frequently in arid environments, resulting in great damage the society and the environment. In Oman, severe WFFs have occurred repeatedly within the last 10 years causing a huge impact on human lives and properties. This paper aims at introducing the framework of an international collaboration project between Japan and Oman for WFF management considering sediment dynamics and climate changes. Four research groups were established: climate change (G1), rainfall-runoff modeling (G2), sediment yield and transport (G3), and sedimentation and infiltration processes (G4). Several field investigations were conducted since 2017 until now. The detailed field survey to assess the deposited sediment in a dry reservoir by using sediment bars, and infiltration test, as well as drone survey were addressed. Some of the preliminary results and findings from the field investigation is discussed. The results show there is an adverse impact of sedimentation clogging on the infiltration process at the reservoirs. Based on the historical rainfall data analysis, there is a systematic increasing trend of the annual average precipitation with remarkable cycles over the MENA region and Oman. The knowledge obtained from this project is expected to be valuable to understanding sediment dynamics at Wadi basins.

To understand the potential roles of terrestrial bamboo on controlling cyanobacterial blooms in aquatic systems, growth rates of the cyanobacterium Microcystis aeruginosa and its competitor algae were examined under different concentrations of bamboo extract. In mono-species cultures with unicellular algal strains, 5.0 g L−1 extract treatment suppressed M. aeruginosa growth, while it had little effect on the growth of green alga Scenedesmus obliquus or promoted the growth of diatom Nitzschia palea. In co-species cultures, the extract treatment increased the effect of S. obliquus and N. palea on the growth of M. aeruginosa. Under the extract treatment with a field-collected M. aeruginosa population, its cell density declined and its colony was etiolated and sank, while co-cultured N. palea increased explosively by invading the colony. These results suggest that bamboo forest stands along banks and artificially installed bamboo poles can affect the aquatic environment for phytoplankton community. Enhancing the growth of competitors, especially diatoms that can invade cyanobacterial colonies, by using extracts or by providing substrates for growth, was suggested to be the major way of the bloom control by bamboo.

Atherosclerosis has been considered as the main cause of morbidity, mortality, and disability worldwide. The first screening for antigen markers was conducted using the serological identification of antigens by recombinant cDNA expression cloning, which has identified adaptor-related protein complex 3 subunit delta 1 (AP3D1) as an antigen recognized by serum IgG antibodies of patients with atherosclerosis. Serum antibody levels were examined using the amplified luminescent proximity homogeneous assay-linked immunosorbent assay (AlphaLISA) using a recombinant protein as an antigen. It was determined that the serum antibody levels against AP3D1 were higher in patients with acute ischemic stroke, transient ischemic attack , diabetes mellitus (DM), cardiovascular disease , chronic kidney disease (CKD), esophageal squamous cell carcinoma (ESCC), and colorectal carcinoma than those in the healthy donors. The area under the curve values of DM, nephrosclerosis type of CKD, and ESCC calculated using receiver operating characteristic curve analysis were higher than that of other diseases. Correlation analysis showed that the anti-AP3D1 antibody levels were highly associated with maximum intima-media thickness, which indicates that this marker reflected the development of atherosclerosis. The results of the Japan Public Health Center-based Prospective Study indicated that this antibody marker is deemed useful as risk factors for AIS.

Anti‐PUF60 autoantibodies are reportedly detected in the sera of patients with dermatomyositis and Sjögren's syndrome; however, little is known regarding its existence in the sera of cancer patients. FIR, a splicing variant of the PUF60 gene, is a transcriptional repressor of c‐myc. In colorectal cancer, there is an overexpression of the dominant negative form of FIR, in which exon 2 is lacking (FIRΔexon2). Previously, large‐scale SEREX (serological identification of antigens by recombinant cDNA expression cloning) screenings have identified anti‐FIR autoantibodies in the sera of cancer patients. In the present study, we revealed the presence and significance of anti‐FIR (FIR/FIRΔexon2) Abs in the sera of patients with esophageal squamous cell carcinoma (ESCC). Our results were validated by an amplified luminescence proximity homogeneous assay using sera of patients with various cancer types. We revealed that anti‐FIRΔexon2 Ab had higher sensitivity than anti‐FIR Ab. Receiver operating characteristic (ROC) analysis was applied for evaluating the use of anti‐FIRΔexon2 Ab as candidate markers such as anti‐p53 Ab and carcinoembryonic antigen, and the highest area under the ROC curve was observed in the combination of anti‐FIRΔexon2 Ab and anti‐p53 Ab. In summary, our results suggest the use of anti‐FIRΔexon2 Ab in combination with the anti‐p53 Ab as a predictive marker for ESCC. The area under the ROC curve was further increased in the advanced stage of ESCC. The value of anti‐FIRΔexon2 autoantibody as novel clinical indicator against ESCC and as a companion diagnostic tool is discussed.

Riverine ponds, which are formed and sustained through sediment erosion and deposition, are key habitats for enhancing biodiversity in river reaches. The objective of this study was to understand the roles of traditional river-training wooden structures called "seigyu" on the formation of ponds on nonvegetated bars. Here, the spatial and temporal patterns of the flow and bedform coupled with pond formations for several flood events were assessed. The surface flow patterns were monitored by an unmanned aerial vehicle (UAV) and evaluated by large-scale particle image velocimetry (LSPIV); the maximum flow velocities were 1.3 m s

Background: Numerous antibody biomarkers have been reported for cancer and atherosclerosis-related diseases. The major complications of atherosclerosis and diabetes mellitus (DM) are acute ischemic stroke (AIS), cardiovascular disease (CVD), and chronic kidney disease (CKD). Cancer development is accompanied by arterial disorders such as angiogenesis and atherosclerosis, and DM is a risk factor for certain cancers. Atherosclerosis-related diseases and cancers are therefore interrelated and could be detected by a common biomarker.Methods: We employed the protein array method for the initial screening of antigens and employed the amplified luminescent proximity homogeneous assay-linked immunosorbent assay (AlphaLISA) to evaluate antibody levels in serum samples.Results: The protein array identified KIAA0513 as an antigen recognized by serum IgG antibodies in the sera of patients with atherosclerosis. We then prepared recombinant glutathione S-transferase-fused KIAA0513 protein. AlphaLISA showed significantly higher serum antibody levels against KIAA0513 protein in patients with AIS, transient ischemic attack, DM, CVD, obstructive sleep apnea syndrome (OSAS), CKD, and solid cancers, such as esophageal, gastric, colon, lung, and breast cancer, than in healthy donors. A receiver operating characteristic (ROC) analysis revealed that the highest areas under the ROC curves of anti-KIAA0513 antibodies were for esophageal cancer, nephrosclerosis-type CKD, and DM. Spearman’s correlation analysis revealed that serum anti-KIAA0513 antibody levels were associated with maximum intima-media thickness and plaque score, which are indices of atherosclerosis and stenosis.Conclusion: Serum anti-KIAA0513 antibody markers appear to be useful for diagnosing AIS, TIA, DM, CVD, OSAS, CKD, and solid cancers and might reflect common arterial alterations leading to atherosclerotic and cancerous diseases.

Background: Atherosclerosis has been considered as the main cause of morbidity, mortality, premature incapacity, and disability worldwide. For early and sensitive diagnosis, development of novel biomarkers is expected and of significant practical importance.Methods: The first screening for antigen markers was conducted using the serological identification of antigens by recombinant cDNA expression cloning (SEREX). Serum antibody levels were examined using the amplified luminescent proximity homogeneous assay-linked immunosorbent assay (AlphaLISA) using a recombinant protein as an antigen.Results: SEREX screening has identified adaptor-related protein complex 3 subunit delta 1 (AP3D1) as an antigen recognized by serum IgG antibodies of patients with atherosclerosis. As per the results of AlphaLISA, it was determined that the serum antibody levels against AP3D1 were higher in patients with acute ischemic stroke (AIS), transient ischemic attack (TIA), diabetes mellitus (DM), cardiovascular disease (CVD), chronic kidney disease (CKD), esophageal squamous cell carcinoma (ESCC), and colorectal carcinoma than those in the healthy donors. The area under the curve values of DM, nephrosclerosis type of CKD, and ESCC calculated using receiver operating characteristic curve analysis were noted to be higher than that of other diseases. Correlation analysis showed that the anti-AP3D1 antibody levels were highly associated with maximum intima-media thickness, which indicates that this marker reflected the development of atherosclerosis. The Japan Public Health Center-based Prospective Study results have verified that the serum antibody levels against AP3D1 showed significantly higher odds ratios with the risk of AIS for persons with the third quartiles and the highest quartiles versus the lowest quartile, indicating that this antibody marker is deemed useful as risk factors for AIS.Conclusions: Serum anti-AP3D1 antibodies, which are broad-spectrum biomarkers of atherosclerotic diseases and digestive organ cancers, could be useful in predicting the onset of AIS.

Effects of substrate and water depth on the physiological status of a submerged macrophyte,Vallisneria natans(Lour.) H. Hara, were determined by measuring biomarkers in leaves and roots, to understand factors limiting the re-establishment ofV. natansin urban eutrophic ponds. Ramets ofV. natanswere grown in the laboratory using aquaria containing water and bottom mud from a eutrophic pond and maintained under sufficient light in an incubator. The growth and chlorophyll-a (Chl-a) content of leaves were greater in aquaria with mud than in those with sand, which was used as the reference substrate. The contents of a peroxidation product (malondialdehyde (MDA)) and three antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD)) in leaves and roots, used as stress biomarkers, changed during the experiment, although differences in these contents between mud and sand were not consistent across the experimental days. To control water depth in the field, ramets ofV. natanswere grown in cages with different substrates (mud and sand) installed at different depths (0.5, 1.2, and 2.0 m) in the pond. The mean light quantum during the experiment decreased with increasing depth, from 79.3 μmol/m2s at 0.5 m to 7.9 μmol/m2s at 2.0 m. The Chl-a content in leaves decreased, whereas the MDA content in both leaves and roots increased with increasing water depth. All enzyme activities increased at the beginning and then decreased to the end of the experiment at 2.0 m depth, suggesting deterioration of enzyme activities due to depth-related stress. The MDA content and CAT activity were higher for sand than for mud, whereas the difference in the growth and the leaf Chl-a content between substrates remained unclear in the pond. On comparing the laboratory and field experiments, the leaf Chl-a content was found to be lower and the MDA content and enzyme activities exhibited sharp increase for ramets grown in the pond, even at 0.5 m depth, when compared with those grown in the aquaria. Our results suggest that the bottom mud of the pond is not the major limiting factor in the re-establishment ofV. natans. Because water depth and light attenuation exerted strong stress onV. natans, shallow areas or measures to improve water transparency are required to promote the introduction ofV. natansin eutrophic ponds for successful restoration in urban areas.

Brahma-related gene 1 (BRG1), an ATPase subunit of the SWItch/sucrose non-fermentable (SWI/SNF) chromatin remodeling complex controls multipotent neural crest formation by regulating epithelial-mesenchymal transition (EMT)-related genes with adenosine triphosphate-dependent chromodomain-helicase DNA-binding protein 7 (CHD7). The expression of BRG1 engages in pre-mRNA splicing through interacting RNPs in cancers; however, the detailed molecular pathology of how BRG1and CHD7 relate to cancer development remains largely unveiled. This study demonstrated novel post-transcriptional regulation of BRG1 in EMT and relationship with FIRΔexon2, which is a splicing variant of the far-upstream element-binding protein (FUBP) 1-interacting repressor (FIR) lacking exon 2, which fails to repress c-myc transcription in cancers. Previously, we have reported that FIR complete knockout mice (FIR−/−) was embryonic lethal before E9.5, suggesting FIR is crucial for development. FIRΔexon2 acetylated H3K27 on promoter of BRG1 by CHIP-sequence and suppressed BRG1 expression post-transcriptionally; herein BRG1 suppressed Snai1 that is a transcriptional suppressor of E-cadherin that prevents cancer invasion and metastasis. Ribosomal proteins, hnRNPs, splicing-related factors, poly (A) binding proteins, mRNA-binding proteins, tRNA, DEAD box, and WD-repeat proteins were identified as co-immunoprecipitated proteins with FIR and FIRΔexon2 by redoing exhaustive mass spectrometry analysis. Furthermore, the effect of FIRΔexon2 on FGF8 mRNA splicing was examined as an indicator of neural development due to impaired CHD7 revealed in CHARGE syndrome. Expectedly, siRNA of FIRΔexon2 altered FGF8 pre-mRNA splicing, indicated close molecular interaction among FIRΔexon2, BRG1 and CHD7. FIRΔexon2 mRNA was elevated in human gastric cancers but not in non-invasive gastric tumors in FIR+/ mice (K19-Wnt1/C2mE x FIR+/−). The levels of FIR family (FIR, FIRΔexon2 and PUF60), BRG1, Snai1, FBW7, E-cadherin, c-Myc, cyclin-E, and SAP155 increased in the gastric tumors in FIR+/− mice compared to those expressed in wild-type mice. FIR family, Snai1, cyclin-E, BRG1, and c-Myc showed trends toward higher expression in larger tumors than in smaller tumors in Gan-mice (K19-Wnt1/C2mE). The expressions of BRG1 and Snai1 were positively correlated in the gastric tumors of the Gan-mice. Finally, BRG1 is a candidate substrate of F-box and WD-repeat domain-containing 7 (FBW7) revealed by three-dimensional crystal structure analysis that the U2AF-homology motif (UHM) of FIRΔexon2 interacted with tryptophan-425 and asparate-399 (WD)-like motif in the degron pocket of FBW7 as a UHM-ligand motif. Together, FIRΔexon2 engages in multi-step post-transcriptional regulation of BRG1, affecting EMT through the BRG1/Snai1/E-cadherin pathway and promoting tumor proliferation and invasion of gastric cancers.

Herein, we summarized the current and future study topics of sediment management using bypass tunnels based on the discussions at the Second International Workshop on Sediment Bypass Tunnels (SBTs) at Kyoto in May 2017. Although reservoir sediment management using bypass tunnels has appeared since the beginning of the 20th century in Kobe, the number of SBTs worldwide is still limited. To promote the installation of SBTs as an effective sediment management measure, it is essential to appropriately evaluate their long-term advantages economically and for the restoration of the aquatic ecosystem. An abrasion model has been developed to predict the abrasion rate of tunnels from the volume of sediment transport. Further, methods to monitor sediment transport in tunnels have advanced. With a significant amount of sediment supply by SBTs, the ecosystems in the downstream reaches of dams can be restored within a few years. A precise rainfall and runoff model for predicting the inflow hydrograph and sediment is essential for the efficient operation of dam gates and SBT (e.g., diverting minimum amount of water for sediment transport). Further studies are needed to clarify the suitable grain size for transportation through SBTs in terms of both the mitigation of tunnel abrasion and promotion of the restoration of downstream ecosystems.

Overexpression of alternative splicing of far upstream element binding protein 1 (FUBP1) interacting repressor (FIR; poly(U) binding splicing factor 60 [PUF60]) and cyclin E were detected in esophageal squamous cell carcinomas (ESCC). Accordingly, the expression of FBW7 was examined by which cyclin E is degraded as a substrate via the proteasome system. Expectedly, FBW7 expression was decreased significantly in ESCC. Conversely, c-myc gene transcriptional repressor FIR (alias PUF60; U2AF-related protein) and its alternative splicing variant form (FIRΔexon2) were overexpressed in ESCC. Further, anticancer drugs (cis-diaminedichloroplatinum/cisplatin [CDDP] or 5-fluorouracil [5-FU]) and knockdown of FIR by small interfering RNA (siRNA) increased cyclin E while knockdown of FIRΔexon2 by siRNA decreased cyclin E expression in ESCC cell lines (TE1, TE2, and T.Tn) or cervical SCC cells (HeLa cells). Especially, knockdown of SAP155 (SF3b1), a splicing factor required for proper alternative splicing of FIR pre-mRNA, decreased cyclin E. Therefore, disturbed alternative splicing of FIR generated FIR/FIRΔexon2 with cyclin E overexpression in esophageal cancers, indicating that SAP155 siRNA potentially rescued FBW7 function by reducing expression of FIR and/or FIRΔexon2. Remarkably, Three-dimensional structure analysis revealed the hypothetical inhibitory mechanism of FBW7 function by FIR/FIRΔexon2, a novel mechanism of cyclin E overexpression by FIR/FIRΔexon2-FBW7 interaction was discussed. Clinically, elevated FIR expression potentially is an indicator of the number of lymph metastases and anti-FIR/FIRΔexon2 antibodies in sera as cancer diagnosis, indicating chemical inhibitors of FIR/FIRΔexon2-FBW7 interaction could be potential candidate drugs for cancer therapy. In conclusion, elevated cyclin E expression was, in part, induced owing to potential FIR/FIRΔexon2-FBW7 interaction in ESCC.

The present study was planned to identify novel serum antibody markers for digestive organ cancers. We have used screening by phage expression cloning and identified novel fourteen antigens in this experiment. The presence of auto-antibodies against these antigens in serum specimens was confirmed by western blotting. As for auto-antibodies against fourteen antigens, AlphaLISA (amplified luminescence proximity homogeneous assay) assay was performed in the sera of gastrointestinal cancers patients to confirm the results. Serum antibody levels against these fourteen recombinant proteins as antigens between healthy donors (HD) and esophageal squamous cell carcinoma (ESCC) patients, gastric cancer (GC), or colon cancer (CC) were compared. The serum levels of all fourteen auto-antibodies were significantly higher in ESCC and GC than those of HD. Among those auto-antibodies, except ECSA2 and CCNL2, were also detected significantly higher levels in CC than those of HD. Receiver operating curve (ROC) revealed similar results except CCNL2 in CC. AUC values calculated by ROC were higher than 0.7 in auto-antibodies against TPI1, HOOK2, PUF60, PRDX4, HS3ST1, TUBA1B, TACSTD2, AKR1C3, BAMBI, DCAF15 in ESCC, auto-antibodies against TPI1, HOOK2, PUF60, PRDX4, TACSTD2, AKR1C3, BAMBI, DCAF15 in GC, and auto-antibodies against TPI1, HOOK2, PUF60 in CC. AUC of the combination of HOOK2 and anti-p53 antibodies in ESCC was observed to be as high as 0.8228. Higher serum antibody levels against ten antigens could be potential diagnostic tool for ESCC. Higher serum antibody levels against eight antigens could be potential diagnostic tool for GC, and serum antibody levels against three antigens could be potential diagnostic tool for CC.

To enhance water quality in eutrophic urban ponds, the effects of a portable water circulation device, equipped with biological oxidation carriers and light-emitting diodes (LEDs), on dissolved oxygen (DO), water transparency, total nitrogen (TN), total phosphorus (TP), chemical oxygen demand (COD), and plankton community in laboratory and field mesocosms were examined. In the laboratory, water quality was monitored in vessels containing water and mud collected from an urban pond, which were subjected to different treatments (control, device without LEDs, and device with LEDs) for 28 days. In the treatments using the device (with and without LEDs), DO increased by aeration, whereas TP decreased by mitigation of mud anoxia. Reductions in TN and the presence of phyto- and zooplankton were observed in those treatments with LEDs, in which periphyton growth was stimulated in the carriers. Under field conditions, water quality of the pond was monitored for 40 days in enclosures. Clear increases in DO and transparency and reductions in TN, TP, and COD were detected in the treatments using the device. Reductions in cyanobacteria and increases in diatoms, green algae, and zooplankton were observed in the treatments with LEDs. Thus, the circulation device effectively improved oxygen condition of the bottom and nutrient flux between sediment and overlying water. In addition, these results suggest that the COD removal and growth of diatoms and green algae by LEDs in carriers play roles on developing periphyton at the bottom and suitable phytoplankton community in the water column.

Four dams in Japan and Switzerland with Sediment Bypass Tunnels (SBT) as a measure against reservoir sedimentation were monitored to analyse the effects of sediment supply on the downstream environment based on up- to downstream differences in geomorphological and biological characteristics. SBT operation times ranged from 93 years at Pfaffensprung and 17 at Asahi to only three years at Solis and no operation at Koshibu. Sediment grain size distribution was monitored, and microhabitats and invertebrates were analysed in terms of richness and composition. Results showed that grain sizes were coarser down- than upstream at dams with newly established SBTs, while they were similar or finer for dams with long SBT operation. Analysis of biotic data revealed that microhabitat and invertebrate richness was low directly below the dam but increased further downstream the longer the SBT operation. Sedentary species dominated at locations where bed conditions were stable, e.g. directly downstream of the dam ...

Sediment replenishment is an effective method for resupplying depleted sediment and detaching overgrown algae in the downstream reaches of a dam. In this study, we used empirical data to examine the effects of sediment replenishment on bed material size and algal biomass in the downstream reaches of the Futase Dam, Chichibu City, Saitama Prefecture, Japan. Assuming that algae detach from bed materials when they are moved by water flow, we calculated the tractive force on the riverbed (τ) and allotted a threshold bed material size in motion (Dcri) for each given τ. The resulting bed material in the downstream reaches of the dam in any year was typically finer than that in the previous year when flooding in the rainy season transported a large volume of sediment. Algal biomass was lower when monthly Dcri exceeded 2 mm, versus when it was less than 2 mm. These results suggest that replenishment of fine bed materials accelerates algal detachment and restricts the accumulation of algal biomass by reducing bed stability.

// Sohei Kobayashi 1 , Tyuji Hoshino 2 , Takaki Hiwasa 3 , Mamoru Satoh 4 , Bahityar Rahmutulla 1, 5 , Sachio Tsuchida 4 , Yuji Komukai 2 , Tomoaki Tanaka 1 , Hisahiro Matsubara 6 , Hideaki Shimada 7 , Fumio Nomura 4 , Kazuyuki Matsushita 1, 8 1 Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Chiba City, Chiba 260-8670, Japan 2 Department of Physical Chemistry, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 260-8675, Japan 3 Department of Biochemistry and Genetics, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan 4 Divisions of Clinical Mass Spectrometry and Clinical Genetics, Chiba University Hospital, Chiba 260-8670, Japan 5 Department of Molecular Oncology, Graduate School of Medicine, Chiba University, Chiba City, Chiba 260-8670, Japan 6 Department of Academic Surgery, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan 7 Department of Gastroenterological Surgery, Toho University Omori Medical Center, Tokyo 143-8541, Japan 8 Department of Laboratory Medicine & Division of Clinical Genetics and Proteomics Chiba University Hospital, Chiba 260-8670, Japan Correspondence to: Kazuyuki Matsushita, email: kmatsu@faculty.chiba-u.jp Keywords: auto-antibodies, cancer biomarker, colorectal cancer, far-upstream element-binding protein-interacting repressor (FIR) = poly(U)-binding-splicing factor (PUF60) Received: June 29, 2016 Accepted: October 12, 2016 Published: October 15, 2016 ABSTRACT Anti-PUF60, poly(U)-binding-splicing factor, autoantibodies are reported to be detected in the sera of dermatomyositis and Sjogren’s syndrome that occasionally associated with malignancies. PUF60 is identical with far-upstream element-binding protein-interacting repressor (FIR) that is a transcriptional repressor of c-myc gene. In colorectal cancers, a splicing variant of FIR that lacks exon2 (FIRΔexon2) is overexpressed as a dominant negative form of FIR. In this study, to reveal the presence and the significance of anti-FIRs (FIR/FIRΔexon2) antibodies in cancers were explored in the sera of colorectal and other cancer patients. Anti-FIRs antibodies were surely detected in the preoperative sera of 28 colorectal cancer patients (32.2% of positive rates), and the detection rate was significantly higher than that in healthy control sera (Mann–Whitney U test, p < 0.01). The level of anti-FIRs antibodies significantly decreased after the operation ( p < 0.01). Anti-FIRs antibodies were detected in the sera of early-stage and/or recurrent colon cancer patients in which anti-p53 antibodies, CEA, and CA19-9 were not detected as well as in the sera of other cancer patients. Furthermore, the area under the curve of receiver operating characteristic for anti-FIRs antibodies was significantly larger (0.85) than that for anti-p53 antibodies or CA19-9. In conclusions, the combination of anti-FIRs antibodies with other clinically available tumor markers further improved the specificity and accuracy of cancer diagnosis.

Large woody debris (LWD) could increase the risk of flooding in the event of heavy rainfall, and fast estimation of the accumulated LWD is crucial for river basin management. We develop a method to automatically detect LWD floating in reservoirs using airborne images, and estimated the total volume of LWD accumulated in a reservoir during a severe flood event in northern Kyushu, Japan, early July 2017. The LWD was successfully extracted from the images based on colour and brightness by using ImageJ. The total volume of the accumulated LWD estimated from images was similar to the LWD volume actually removed from the reservoir by the end of October. Most LWD pieces were concentrated in the upstream-side of the reservoir, and a longitudinal variation in density and length of LWD pieces was also evident.

Background The clinical course of ulcerative colitis (UC) is characterized by repeated episodes of relapse and remission. We hypothesized that biomarkers that help distinguish refractory UC patients who are in remission using strong anti-immunotherapy could contribute in preventing the overuse of corticosteroids for treatment. Here, we clarified novel autoantibodies for UC patients in remission as clinical indicators to distinguish between refractory and non-refractory UC. Methods Antigen proteins recognized by serum antibodies of patients with UC in remission were screened using the protein array method. To validate the results, AlphaLISA was used to analyze the serum antibody titers with candidate protein antigens. Serum samples from 101 healthy controls, 121 patients with UC, and 39 patients with Crohn's disease were analyzed. Results Of 66 candidate protein antigens screened by ProtoArray™, six were selected for this study. The serum titers of anti-poly ADP-ribose glycohydrolase (PARG), anti-transcription elongation factor A protein-like 1, and anti-proline-rich 13 (PRR13) antibodies were significantly higher in patients with UC than in healthy controls. Anti-PARG and anti-PRR13 antibody titers were significantly higher in patients with refractory UC than in patients with non-refractory UC. There were no significant differences in any antibody titer between the active and remission phases. Conclusions The serum titers of anti-PARG, anti-transcription elongation factor A protein-like 1, and anti-PRR13 antibodies were elevated in patients with UC. Anti-PARG and anti-PRR13 antibody titers may be novel clinical indicators for detecting refractory UC in patients in remission.

Sediment replenishment is a technique employed to prevent sediment deficiency in riverbeds The distribution of sediment in a riverbed is affected by the volume of replenished sediment in addition to the flood discharge We examine the effects of flood discharge on the riverbed material distributions by analyzing a case of sediment replenishment in the Futase Dam in Saitama Prefecture Japan Specifically we compared data from to on the annual volume of flushed sediment Sf the annual maximum flood discharge Qmax and the proportion of particles less than mm in diameter P lt mm Results showed that the annual P lt mm at the downstream site may decrease following heavy flooding i e Qmaxreaches a ten year high possibly as a result of fine particles on the riverbed being carried away by flood water Alternatively the area covered by the replenishing sediment may be limited when Qmax is at a ten year low Thus in the case of the Futase Dam a moderate degree of flooding may facilitate sediment transport at the downstream reach and distribute fine particles on the riverbed

Flash floods in wadi systems discharge large volumes of water to either the sea or the desert areas after high-intensity rainfall events. Recently, wadi flash floods have frequently occurred in arid regions and caused damage to roads, houses, and properties. Therefore, monitoring and quantifying these events by accurately measuring wadi discharge has become important for the installation of mitigation structures and early warning systems. In this study, image-based methods were used to measure surface flow velocities during a wadi flash flood in 2018 to test the usefulness of large-scale particle image velocimetry (LSPIV) and space&ndash, time image velocimetry (STIV) techniques for the estimation of wadi discharge. The results, which indicated the positive performance of the image-based methods, strengthened our hypothesis that the application of LSPIV and STIV techniques is appropriate for the analysis of wadi flash flood velocities. STIV is suitable for unidirectional flow velocity and LSPIV is reliable and stable for two-dimensional measurement along the wadi channel, the direction of flow pattern which varies with time.

We compared biomass and community structure of macroinvertebrates among 3 flow zones (deep, rapid, flat) of riffles at 3 sites in a gravel-bed river. We evaluated bed stability in these zones with a 2-dimensional hydrodynamic simulation over a range of discharge levels. Deep zones had higher flow velocity and coarser bed materials than other zones. Rapid zones were shallower with higher flow velocity than flat zones. The probability of bed movement was greatest in rapid zones and was lowest in deep zones based on bed shear stress and the size of bed materials. Total macroinvertebrate biomass was dominated by filterer insects and was highest in deep zones and lowest in rapid zones across the sites. This trend was most conspicuous for taxa that build retreats on stones, such as net-spinning caddisflies, which have a sessile life form and prefer stable environments. The trend was less apparent for taxa that move freely on the bed, such as baetid and heptageniid mayflies. The macroinvertebrate communi...

To understand the impacts of debris flows on the distribution of an amphipod with limited dispersal ability in the context of stream networks, we surveyed the presence of Gammarus nipponensis in 87 headwater streams with different legacies of debris flow occurrence within an 8.5-km2 mountain catchment. The amphipod was present in only 7% of the streams impacted by debris flows after 1977; in contrast, it was present in 69% of the streams that had older or no debris flow occurrence. The absence of the amphipod in certain headwater streams did not appear to be related to water chemistry because pH and calcium concentrations differed little among streams within the catchment. In addition, survival rates of individuals incubated in streams with the amphipod present and absent did not differ significantly. Debris flows appeared to displace amphipod populations, and the absence of amphipods in streams for more than 30 years after debris flow occurrence suggests that considerable time is required for the recovery of populations. Because of geographic isolation from the source of colonists, headwater streams in the uppermost sections of the catchment and those indirectly connecting to the main stream via tributaries appear to be at greater disadvantage for receiving colonists from other areas and thus population recovery. Copyright © 2012 John Wiley & Sons, Ltd.

Particulate organic matter (POM) plays an important role in nutrient dynamics in river ecosystems, but little is known about changes in the origin and quality of POM in relation to physical and seasonal changes along rivers. Using stable isotope and stoichiometric analyses, we investigated the changes in origin and quality of POM of three different size fractions (fine [FPOM], 1.2–100 μm; medium [MPOM], 100–250 μm; and coarse [CPOM], 250–1,000 μm) at 14 sites along a gravel-bed river over four seasons. FPOM and MPOM accounted for 90% of all POM at all study sites. At each site, the δ13C level was lower for FPOM (range: −29.0 to −21.1‰) than for MPOM (−26.9 to −17.2‰) and CPOM (−27.5 to −16.3‰). The C:N ratio was lower for FPOM (6.9–15.6) than for MPOM (6.3–17.4) and CPOM (5.7–27.1). The contribution of periphyton to POM of all size fractions had a tendency to increase downstream, though the trend was less clear and varied seasonally for MPOM and CPOM between sites in middle and downstream reaches. Contrastively, the C:N ratio in all size fractions of POM consistently decreased downstream. The downstream decrease in the C:N ratio of POM can be partly explained by the increase in the contribution of periphyton, which seems to be associated with increased discharge and enhanced periphyton dislodgement, especially in winter. In addition, an increase in bacterial biomass associated with the greater nutrient availability in pool areas is another possible reason for the decrease in the C:N ratio of POM downstream.

For perennial forage grasses, tillering and shoot elongation are important developmental events to determine grassland production. To thoroughly identify and characterize quantitative trait locus (QTL) regions controlling the events, a perennial ryegrass (Lolium perenne L.) mapping population comprising 188 F 2 individuals was evaluated for tiller number (TN), plant length (PL), and leaf length (LL) over 12 cuts in replicated field and pot experiments in Sapporo, Japan, during 2 yr. Quantitative trait loci were repeatedly detected in 16 genomic regions. Based on the QTL, 10 regions affected both shoot elongation (PL and LL) and tillering (TN), whereas only six regions had a single effect on either event. Five QTL regions mapped on linkage groups 1,2,5, and 6, explaining at maximum 34 to 56% phenotypic variance, likely contain key genes for plant development. Profiles (e.g., direction and magnitude) of QTL effects differed greatly among most regions, indicating a large diversity among the genes involved. Statistical tests revealed significant interaction effects of genotype with growth condition and cut for all QTL regions. The QTL positions were compared with those reported for ryegrass and rice, suggesting the presence of genes identical to the current population and semiwild perennial for-age grasses. The current QTL analysis indicated that the control of ryegrass plant development involves diverse, unique genes whose effects are enhanced or suppressed by growth environment and foliage cutting.

Summary 1. To understand longitudinal changes in the trophic base of benthic macroinvertebrates from mountain to lowland river sections, we investigated carbon stable isotopic compositions (δ13C) of macroinvertebrates and their food resources in riffles for four seasons at 14 sites along the main stem of the Toyo River, Japan. 2. At each site, δ13C was usually highest or nearly highest for periphyton (epilithic biofilm) and was lowest for transported leaf materials. Among macroinvertebrate groups, grazers usually had higher δ13C values than filterers or predators. 3. During all seasons, δ13C of periphyton and all macroinvertebrate groups increased downstream from mountain to upland sections, but decreased downstream from upland to lowland sections. In addition, the difference between grazer δ13C and filterer δ13C decreased from mountain to upland sections, but increased from upland to lowland sections. 4. The observed changes in δ13C of periphyton and macroinvertebrates from mountain to upland sections agree with previous reports: the δ13C of periphyton and consumers increased with stream size and productivity. The decrease in δ13C of periphyton and macroinvertebrates from upland to lowland sections has not been reported previously, and this may have resulted from an increased importance of terrestrial detritus relative to periphyton production in the lowland section, where riffles were infrequent and pools dominated the reach. 5. A simple mixing model of δ13C showed that grazers rely mostly on periphyton at all sites, whereas the importance of periphyton for filterers changed longitudinally increasing from mountain to upland sections and decreasing from upland to lowland sections. This longitudinal trend for filterers is possibly associated with the changes in the availability or quality of terrestrial detritus in transported particulate organic matter. 6. Longitudinal changes in the relative importance of autochthonous production and allochthonous detritus appear to be reflected in δ13C of riffle benthic communities. The longitudinal changes were not monotonic, and specific reach characteristics may be responsible for the greater importance of allochthonous detritus in mountain and lowland sections.

To analyze how silicon (Si) fertilizer improves plant growth under water stress, we investigated the growth and root responses of sorghum seedlings to Si application. Seedlings were grown hydroponically at two Si levels (0 and 1.78 mM) and under two water stress conditions simulated with polyethylene glycol. The reduction in dry weight due to stress was alleviated by Si application, accompanied by an increase in root water uptake. Silicon application decreased the osmotic potential of the roots without affecting their water content, showing that osmotic adjustment occurred to increase water uptake. An assessment of root solutes suggested that soluble sugar and amino acids (alanine and glutamic acid) were osmolytes responsible for this adjustment. Root anatomical traits related to water transport were not affected by the Si application. These results improved our understanding of the physiological mechanisms that underlie the Si-induced increases in sorghum growth and water uptake under water stress.

Purpose Human serum and plasma are often used as clinical specimens in proteomics analyses, and peptidome profiling of human serum is a promising tool for identifying novel disease-associated biomarkers. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is widely used for peptidomic biomarker discovery. Careful sample collection and handling are required as either can have a profound impact on serum peptidome patterns, yet the effects of preanalytical variables on serum peptidome profiles have not been completely elucidated. The present study investigated the effects of preanalytical variables, including storage temperature, duration (up to 12 months), and thawing methods, on MALDI-TOF MS-based serum peptidome patterns. Experimental design Aliquots of serum samples were pretreated with weak cation exchanger magnetic beads using an automated ClinProtRobot system and then analyzed by MALDI-TOF MS. Results A number of significant differences in peak intensities were observed depending on sample processing variables. Conclusions and clinical relevance These peaks can be used as sample quality markers to assess the effects of long-term storage on serum peptidome profiles using MALDI-TOF MS.

We examined short-term responses of macroinvertebrate drift associated with experimental sediment flushing in a headwater. Increases in the drifting abundances of Ephemeroptera and Plecoptera coincided with increases in bed load yield rather than peaks in discharge or suspended sediment concentrations. The approach and arrival of a sediment wave may provide a physical cue that initiates the escape of benthic macroinvertebrates. Because fine bed load sediments, with diameters

河川における瀬の物理特性と底生動物現存量の関係を理解するため，愛知県豊川において流程間（下流から平野部，峡谷部，山間部，山地部）で瀬の物理特性と底生動物群集の比較を行った。河床礫は山間部や山地部の地点で大きかったが，年最大流量で動きうる礫径に対する実際の河床礫径の比は平野部の地点で大きかった。一方，平水時の平均流速は峡谷部と山間部の地点で大きい傾向にあった。年間平均の全底生動物現存量は平野部で最も大きく，次いで山間部で大きかった。摂食機能群別に現存量をみると，付着物食者は平野部，濾過食者は平野部と山間部で大きい一方，捕食者は山間部と山地部で大きかった。底生動物を河床中の生息拠点と移動・固着性を基に河床生息型区分を行うと，平野部で多いのは礫面-固着巣型や礫下砂-固着巣型などいずれも安定した環境を好むグループ，一方，山間部と山地部で多いのは礫間-固着巣型，礫間-自由型など礫間の空隙を利用するグループであった。これらに基づき，底生動物現存量に貢献し群集を特徴づけている瀬の物理特性は，平野部では河床安定性，山間部では礫の大きさに伴う礫間空隙量と考えられた。