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ASOs suppress ARL4C mRNA expression in vitro.

ADP-ribosylation factor-like 4c (ARL4C) is identified as a small GTP-binding protein, which is expressed by Wnt and EGF signaling and plays an important role in tubulogenesis of cultured cells and the ureters. ARL4C is little expressed in adult tissues, but it is highly expressed in lung cancer and colorectal cancer and shown to represent a molecular target for cancer therapy based on siRNA experiments. This study revealed that ARL4C is highly expressed in primary hepatocellular carcinoma (HCC) tumors and colorectal cancer liver metastases, and that ARL4C expression is associated with poor prognosis for these cancers. Chemically modified antisense oligonucleotides (ASO) against ARL4C effectively reduced ARL4C expression in both HCC and colorectal cancer cells and inhibited proliferation and migration of these cancer cells in vitro. ARL4C ASOs decreased the PIK3CD mRNA levels and inhibited the activity of AKT in HCC cells, suggesting that the downstream signaling of ARL4C in HCC cells is different from that in lung and colon cancer cells. In addition, subcutaneous injection of ARL4C ASO was effective in reducing the growth of primary HCC and metastatic colorectal cancer in the liver of immunodeficient mice. ARL4C ASO accumulated in cancer cells more efficiently than the surrounding normal cells in the liver and decreased ARL4C expression in the tumor. These results suggest that ARL4C ASO represents a novel targeted nucleic acid medicine for the treatment of primary and metastatic liver cancers.

The sequences of ARL4C ASOs (Table S1), primers (Table S2) and siRNA (Table S3). Relationship between ARL4C expression and clinicopathological facrors of HCC and CRC cases (Table S4). Univariate analysis of relapse-free survival of HCC and CRC (Table S5). mRNA levels of candidate off-target genes for ARL4C ASO (Table S6).

Wnt5a is a representative ligand that activates the β-catenin–independent pathway in Wnt signaling. It was reported that the expression of Wnt5a in human gastric cancer is associated with aggressiveness and poor prognosis and that knockdown of Wnt5a reduces the ability of gastric cancer cells to metastasize in nude mice. Therefore, Wnt5a and its signaling pathway might be important targets for the therapy of gastric cancer. The aim of this study was to examine whether an anti-Wnt5a antibody affects metastasis of gastric cancer cells. One anti-Wnt5a polyclonal antibody (pAb5a-5) inhibited migration and invasion activities in vitro of gastric cancer cells with a high expression level of Wnt5a. Previously, it was shown that Wnt5a induces the internalization of receptors, which is required for Wnt5a-dependent activation of Rac1. pAb5a-5 inhibited Wnt5a-dependent internalization of receptors, thereby suppressed Wnt5a-dependent activation of Rac1. Laminin γ2 is one of target genes of Wnt5a signaling and Rac1 was involved in its expression. pAb5a-5 also inhibited Wnt5a-dependent expression of laminin γ2. In an experimental liver metastasis assay, gastric cancer cells were introduced into the spleens of nude mice. Laminin γ2 was required for liver metastatic ability of gastric cancer cells in vivo. Furthermore, intraperitoneal injection of pAb5a-5 inhibited the metastatic ability of gastric cancer cells. These results suggest that an anti-Wnt5a antibody was capable of suppressing Wnt5a-dependent internalization of receptors, resulting in the prevention of metastasis of gastric cancer cells by inhibiting the activation of Rac1 and the expression of laminin γ2. Mol Cancer Ther; 11(2); 298–307. ©2011 AACR.

Dickkopf1 (DKK1) was originally identified as a secreted protein that antagonizes Wnt signaling. Although DKK1 is essential for the developmental process, its functions in postnatal and adult life are unclear. However, evidence is accumulating that DKK1 is involved in tumorigenesis in a manner unrelated to Wnt signaling. In addition, recent studies have revealed that DKK1 may control immune reactions, although the relationship of this to Wnt signaling is unknown. Other DKK family members, DKK2-4, are likely to have their own functions. Here, we review the possible novel functions of DKKs. We summarize the characteristics of receptors of DKKs and the signaling mechanisms through DKKs and their receptors, provide evidence showing that DKKs are involved in tumor aggressiveness independently of Wnt signaling, and emphasize promising cancer therapies targeting DKKs and receptors. Lastly, we discuss various physiological and pathological processes controlled by DKKs.

Growth regulation by estrogen in breast cancer 1 (GREB1) is involved in hormone-dependent and -independent tumor development (e.g., hepatoblastoma). In this study, we found that a GREB1 splicing variant, isoform 4 (Is4), which encodes C-terminal half of full-length GREB1, is specifically expressed via microphthalmia-associated transcription factor (MITF) in melanocytic melanoma, and that two MITF-binding E-box CANNTG motifs at the 5’-upstream region of GREB1 exon 19 are necessary for GREB1 Is4 transcription. MITF and GREB1 Is4 were strongly co-expressed in approximately 20% of the melanoma specimens evaluated (17/89 cases) and their expression was associated with tumor thickness. GREB1 Is4 silencing reduced melanoma cell proliferation in association with altered expression of cell proliferation-related genes in vitro. In addition, GREB1 Is4 targeting by antisense oligonucleotide (ASO) decreased melanoma xenograft tumor formation and GREB1 Is4 expression in a BRAFV600E; PTENflox melanoma mouse model promoted melanoma formation, demonstrating the crucial role of GREB1 Is4 for melanoma proliferation in vivo. GREB1 Is4 bound to CAD, the rate-limiting enzyme of pyrimidine metabolism, and metabolic flux analysis revealed that GREBI Is4 is necessary for pyrimidine synthesis. These results suggest that MITF-dependent GREB1 Is4 expression leads to melanoma proliferation and GREB1 Is4 represents a new molecular target in melanoma.

There are currently no treatments for salivary gland diseases, making it vital to understand signaling mechanisms operating in acinar and ductal cells so as to develop regenerative therapies. To date, little work has focused on elucidating the signaling cascades controlling the differentiation of these cell types in adult mammals. To analyze the function of the Hippo-TAZ/YAP1 pathway in adult mouse salivary glands, we generated adMOB1DKO mice in which both MOB1A and MOB1B were TAM-inducibly deleted when the animals were adults. Three weeks after TAM treatment, adMOB1DKO mice exhibited smaller submandibular glands (SMGs) than controls with a decreased number of acinar cells and an increased number of immature dysplastic ductal cells. The mutants suffered from reduced saliva production accompanied by mild inflammatory cell infiltration and fibrosis in SMGs, similar to the Sjogren's syndrome. MOB1-deficient acinar cells showed normal proliferation and apoptosis but decreased differentiation, leading to an increase in acinar/ductal bilineage progenitor cells. These changes were TAZ-dependent but YAP1-independent. Biochemically, MOB1-deficient salivary epithelial cells showed activation of the TAZ/YAP1 and β-catenin in ductal cells, but reduced SOX2 and SOX10 expression in acinar cells. Thus, Hippo-TAZ signaling is critical for proper ductal and acinar cell differentiation and function in adult mice.

Most cancer cells are exposed to altered extracellular environments, such as an increase in extracellular matrix (ECM) stiffness and soluble signals consisting of growth factors and cytokines. It is therefore conceivable that changes in tumor extracellular environments affect tumor cell behavior. The Hippo pathway reportedly responds to the extracellular environment and regulates the nuclear localization of the transcription co-activator, yes-associated protein (YAP)/transcriptional co-activator with PDZ-binding motif (TAZ). Inactivation of the Hippo pathway with nuclear translocation of YAP/TAZ stimulates cell proliferation. Its pathway also regulates gene expression, but the precise molecule(s) meditating the cell-proliferating effect of YAP signaling on oral squamous cell carcinoma (OSCC) is unclear. First, we examined the effects of YAP signaling on OSCC tumorigenesis. Loss-of-function experiments using siRNA or an inhibitor, and immunohistochemical analyses of tissue specimens obtained from OSCC patients demonstrated that YAP signaling was involved in OSCC cell proliferation. Second, we identified Piezo-type mechanosensitive ion channel component 1 (PIEZO1), a Ca2+ channel, as a transcriptional target of YAP signaling and showed that elevated PIEZO1 was required for PIEZO1 agonist-dependent Ca2+ entry and cell proliferation in OSCC cells. Experiments using three-dimensional and suspension culture revealed that PIEZO1 was involved in OSCC cellular growth. Finally, YAP overexpression in the nucleus and/or cytoplasm was immunohistochemically detected in tumor lesions with frequent expression of both PIEZO1 and Ki-67, but not in non-tumor regions of OSCC specimens. These results suggest that the YAP/PIEZO1 axis promotes OSCC cell growth. © 2020 The Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Lung adenocarcinoma is the most common histological type of lung cancer and is classified into adenocarcinoma in situ (AIS), minimally invasive adenocarcinoma (MIA) and invasive adenocarcinoma (IA). Atypical adenomatous hyperplasia (AAH) lesions are possible precursors to adenocarcinoma. However, the mechanism underlying the stepwise continuum of lung adenocarcinoma is unclear. In this study, the involvement of ADP‐ribosylation factor (ARF)‐like (ARL) 4C (ARL4C), a member of the small GTP‐binding protein family, in the progression of lung adenocarcinoma and the possibility of ARL4C as a molecular target for lung cancer therapy were explored. ARL4C was frequently expressed in AAH and ARL4C expression in immortalized human small airway epithelial cells promoted cell proliferation and suppressed cell death. In addition, ARL4C was expressed with increased frequency in AIS, MIA and IA in a stage‐dependent manner, and the expression was correlated with histologic grade, fluorine‐18 fluorodeoxyglucose uptake and poor prognosis. An anti–sense oligonucleotide (ASO) against ARL4C (ARL4C ASO‐1316) inhibited RAS‐related C3 botulinum toxin substrate activity and nuclear import of Yes‐associated protein and transcriptional coactivator with PDZ‐binding motif, and suppressed in vitro proliferation and migration of lung cancer cells with KRAS or epidermal growth factor receptor (EGFR) mutations. In addition, transbronchial administration of ARL4C ASO‐1316 suppressed orthotopic tumor formation induced by these cancer cells. Thus, ARL4C is involved in the initiation of the premalignant stage and is associated with the stepwise continuum of lung adenocarcinoma. ARL4C ASO‐1316 would be useful for lung adenocarcinoma patients expressing ARL4C regardless of the KRAS or EGFR mutation., ARL4C is involved in the initiation of the premalignant stage and is associated with the stepwise continuum of lung adenocarcinoma. Transbronchial administration of ARL4C ASO suppressed orthotopic tumor formation of lung cancer cells.

Neurofibromatosis type 2 (NF2) gene, a tumor suppressor gene located on chromosome 22q12.2, is frequently abnormal in mesothelioma. Recent studies have revealed the effectiveness of diagnostic assays for differentiating malignant pleural mesothelioma from reactive mesothelial hyperplasia. These include detection of homozygous deletion of the 9p21 locus by fluorescence in situ hybridization (FISH) (9p21 FISH), loss of expression of BAP1 as detected by immunohistochemistry, and loss of expression of methylthioadenosine phosphorylase (MTAP) as detected by immunohistochemistry. However, the application of FISH detection of NF2 gene deletion (NF2 FISH) in differentiation of malignant pleural mesothelioma from reactive mesothelial hyperplasia has not been fully evaluated. In this study, we investigated whether NF2 FISH, either alone or in a combination with other diagnostic assays (9p21 FISH, MTAP immunohistochemistry, and BAP1 immunohistochemistry), is effective for distinguishing malignant pleural mesothelioma from reactive mesothelial hyperplasia. This study cohort included malignant pleural mesothelioma (n = 47) and reactive mesothelial hyperplasia cases (n = 27) from a period between 2001 and 2017. We used FISH to examine deletion status of NF2 and 9p21 and immunohistochemistry to examine expression of MTAP and BAP1 in malignant pleural mesothelioma and in reactive mesothelial hyperplasia. Hemizygous NF2 loss (chromosome 22 monosomy or hemizygous deletion) was detected in 25 of 47 (53.2%) mesothelioma cases. None of the mesothelioma cases showed homozygous NF2 deletion. Hemizygous NF2 loss showed 53.2% sensitivity and 100% specificity in differentiating malignant pleural mesothelioma from reactive mesothelial hyperplasia. A combination of NF2 FISH, 9p21 FISH, and BAP1 immunohistochemistry yielded greater sensitivity (100%) than that detected for either diagnostic assay alone (53.2% for NF2 FISH, 78.7% for 9p21 FISH, 70.2% for MTAP immunohistochemistry, or 57.4% for BAP1 immunohistochemistry). Thus, NF2 FISH in combination with other diagnostic assays is effective for distinguishing malignant pleural mesothelioma from reactive mesothelial hyperplasia.

Most human malignant tumor cells arise from epithelial tissues, which show distinctive characteristics, such as polarization, cell-to-cell contact between neighboring cells, and anchoring to a basement membrane. When tumor cells invaginate into the stroma, the cells are exposed to extracellular environments, including the extracellular matrix (ECM). Increased ECM stiffness has been reported to promote cellular biological activities, such as excessive cellular growth and enhanced migration capability. Therefore, tumorous ECM stiffness is not only an important clinical tumor feature but also plays a pivotal role in tumor cell behavior. Transient receptor potential vanilloid 4 (TRPV4), a Ca2+-permeable nonselective cation channel, has been reported to be mechano-sensitive and to regulate tumorigenesis, but the underlying molecular mechanism in tumorigenesis remains unclear. The function of TRPV4 in oral squamous cell carcinoma (OSCC) is also unknown. The current study was conducted to investigate whether or not TRPV4 might be involved in OSCC tumorigenesis. TRPV4 mRNA levels were elevated in OSCC cell lines compared with normal oral epithelial cells, and its expression was required for TRPV4 agonist-dependent Ca2+ entry. TRPV4-depleted tumor cells exhibited decreased proliferation capabilities in three-dimensional culture but not in a low-attachment plastic dish. A xenograft tumor model demonstrated that TRPV4 expression was involved in cancer cell proliferation in vivo. Furthermore, loss-of-function experiments using siRNA or an inhibitor revealed that the TRPV4 expression was required for CaMKII-mediated AKT activation. Immunohistochemical analyses of tissue specimens obtained from 36 OSCC patients showed that TRPV4 was weakly observed in non-tumor regions but was strongly expressed in tumor lesions at high frequencies where phosphorylated AKT expression was frequently detected. These results suggest that the TRPV4/CaMKII/AKT axis, which might be activated by extracellular environments, promotes OSCC tumor cell growth.

Homozygous deletion (homo-d) of the cyclin-dependent kinase inhibitor 2A (CDKN2A) gene is frequently found in malignant pleural mesothelioma (MPM). Fluorescence in situ hybridization (FISH) is commonly used to detect chromosomal deletion, and sometimes reveals more frequent heterozygous deletion (hetero-d) compared with homo-d. In clinical practice, such CDKN2A FISH results belong to the ‘borderline’ homo-d rate, which makes it difficult to definitively diagnose MPM. Microdeletion, [

Rehabilitation is an important stage in mining operations for environmental conservation. However, the shortage of topsoil makes it difficult to achieve rehabilitation in open-cast coal mines. Securing topsoil by mixing soil with fly ash (FA), which is treated as an industrial waste, is expected to solve this issue in coal mines. While mixing soil with FA makes it possible to secure the topsoil and treat industrial waste simultaneously, the high alkalinity of FA and the dissolution of heavy metals from FA may inhibit plant growth. This study investigated the effects of FA in the topsoil on plant growth via vegetation tests with simulated topsoil mixed with FA using Acacia mangium, a species of flowering tree: the FA mixing ratios were set to 0%, 20%, 40%, 60%, and 100%. The growth of Acacia mangium was inhibited with increasing FA mixing ratio, especially from 60% to 80%. However, the growth rate of Acacia mangium in an FA mixing ratio of 100% was nearly comparable to that in a mixing ratio of 40%. Furthermore, there were no effects of the physical characteristics and pH conditions in the topsoil on the plant growth at any of the mixing ratios; meanwhile, the accumulated concentration of Al in the plant body increased significantly at an FA mixing ratio of 60%–80%. This suggests that the accumulation of Al, which inhibits plant growth, including root growth and its functions, in the plant body inhibited the growth of Acacia mangium. Therefore, the most important aspect in terms of rehabilitation concerning the use of FA for securing topsoil is not the mixing ratio of FA but the amount of Al in the FA and the accumulation of Al in the plant body., Rekultywacja jest ważnym etapem w eksploatacji górniczej wzakresie ochrony środowiska. Brak wierzchniej warstwy gleby utrudnia osiągnięcie efektu rekultywacji w kopalniach odkrywkowych. Oczekuje się, że zabezpieczenie wierzchniej warstwy gleby przez zmieszanie gleby z popiołem lotnym (FA), który jest traktowany jako odpad przemysłowy, rozwiąże ten problem w odkrywkowych kopalniach węgla. Podczas mieszania gleby z FA możliwe jest jednoczesne zabezpieczenie wierzchniej warstwy gleby i utylizacja odpadów przemysłowych. Jednak wysoka alkaliczność FA i rozpuszczanie metali ciężkich z FA może hamować wzrost roślin. W artykule przedstawiono wpływ dodatku FA do wierzchniej warstwy gleby na wzrost roślin poprzez testy roślinności z symulowaną wierzchnią warstwą gleby zmieszaną z FA przy użyciu Acacia mangium i gatunków drzew kwitnących. Proporcje mieszanek FA zostały określone na 0%, 20%, 40%, 60% i 100%. Wzrost Acacia Mangium był hamowany wraz ze wzrostem zawartości FA, zwłaszcza od 60% do 80%. Jednak tempo wzrostu w mieszance FA w proporcji 40% było praktycznie takie jak bez dodatku FA. Ponadto nie zaobserwowano wpływu właściwości fizycznych i warunków pH w wierzchniej warstwie gleby na wzrost roślin w żadnej mieszance. Oznacza to, że skumulowane stężenie Al w roślinie znacznie wzrosło dla mieszanek z dodatkiem FA 60% -80%. Sugeruje to, że nagromadzenie Al, które hamuje wzrost roślin, w tym wzrost korzeni i jego funkcje, w organizmie rośliny hamuje wzrost Acacia Mangium. Dlatego najważniejszym aspektem rekultywacji z zastosowaniem FA do zabezpieczania wierzchniej warstwy gleby nie jest wielkość dodatku FA, lecz ilość Al w FA i akumulacja Al w roślinie.

The design and development for a compact 1 GHz (23.5 T)-class NMR magnet are presented. The magnet uses inner coils wound with Ni alloy-reinforced Bi-2223 conductors in combination with LTS outer coils. The series-connected coils are operated in a power supply-driven mode. The total magnet size is very compact, and is comparable to that of a currently commercialized 600-MHz (14.1 T) NMR magnet. This owes to a high current density operation of the Bi-2223 inner coils, which contribute as much as 53% of the total magnetic field. This paper investigated the following properties of the Bi-2223 inner coils: First, joint resistance under external magnetic fields; second, tensile stress tolerances of the Bi-2223 conductor under bending condition; third, high compressive stress on Bi-2223 coils; fourth, the effect of screening current-induced magnetic field on the field homogeneity and stability; and fifth, the maximum permissible Joule heating to avoid a thermal runaway. Two Bi-2223 inner coils for the magnet were built and tested in liquid nitrogen, which provided important knowledge for the magnet excitation.

The β-catenin mutation is frequently observed in hepatoblastoma (HB), but the underlying mechanism by which Wnt/β-catenin signaling induces HB tumor formation is unknown. Here we show that expression of growth regulation by estrogen in breast cancer 1 (GREB1) depends on Wnt/β-catenin signaling in HB patients. GREB1 is localized to the nucleus where it binds Smad2/3 in a competitive manner with p300 and inhibits TGFβ signaling, thereby promoting HepG2 HB cell proliferation. Forced expression of β-catenin, YAP, and c-Met induces HB-like mouse liver tumor (BYM mice), with an increase in GREB1 expression and HB markers. Depletion of GREB1 strongly suppresses marker gene expression and HB-like liver tumorigenesis, and instead enhances TGFβ signaling in BYM mice. Furthermore, antisense oligonucleotides for GREB1 suppress the formation of HepG2 cell-induced tumors and HB-like tumors in vivo. We propose that GREB1 is a target molecule of Wnt/β-catenin signaling and required for HB progression., The mechanisms promoting hepatoblastoma (HB) progression through Wnt/β-catenin signaling are unclear. Here, the authors show that the Wnt/ β-catenin axis induces GREB1 expression and nuclear localization, and suppresses TGFβ pathway, and propose GREB1 as a therapeutic target in HB.

We are developing a cryocooled (RE)Ba2Cu3O7 (REBCO; RE = rare earth) persistent-mode superconducting magnet system that generates 1 T with multiple superconducting joints. In parallel to the design of the system, we have investigated transport characteristics of the REBCO superconducting joints. Critical currents (Ic) were measured as a function of magnetic field and temperature. Two joint samples with Ic of 107 and 53 A at 77 K, self-field were measured. The first one showed Ic of 357 A at 50 K and 0.1 T (magnetic field is perpendicular to the tape surface). Ic of the REBCO joint samples showed remarkable hysteresis. The anisotropy of Ic changed with temperature. Ic at B perpendicular to the tape were higher than those at B parallel to the tape at temperatures below 20 K. The random pinning is possibly the dominant pinning mechanism in the whole temperatures for the REBCO superconducting joints in magnetic fields perpendicular to c-axis.

The technological development for a 3-T high-temperature superconducting (HTS) magnetic resonance imaging (MRI) magnet is underway. The 3-T HTS MRI magnet is imaged such that RE(= Rare Earth)Ba 2 Cu 3 O 7-δ (REBCO) coils are installed and cooled using a cryocooler between 30 and 50 K without liquid helium refrigerant. REBCO MRI magnets are expected to maintain the magnetic field with stability better than 0.1 ppm/h with superconducting joints for a persistent-current mode. The resistance in a joint must be suppressed less than 1 × 10 -12 Ω to achieve the expected stability. The resistance of REBCO joint was estimated measuring the decay rate of trapped magnetic field in a REBCO single-turn loop including a joint. An external magnetic field of up to 1 T was applied to the joint in the STL at temperatures of 35 K and 50 K. The order of resistance in the REBCO joint was less than 1 × 10 -13 Ω.

Soil erosion is one of the major environmental problems in open-cut mines in tropical regions. It causes negative impacts including the removal of nutrient-rich topsoil, destroys aquatic habitat, dam and pond siltation, clogs river by deposition of sediment, and causes water pollution in the rehabilitation process. Soil texture is an important factor to affect soil erosion. In this study, artificial rainfall experiment in the laboratory scale was conducted to clarify the mechanism of soil erosion under the different soil composition and to discuss the methods for minimizing soil erosion. The obtained results showed that the soil seal generated due to the presence of fine particle under high rainfall intensity is the main contributor to accelerate the soil erosion. Additionally, the surface coverage by the cover crops is the most effective measure to reduce soil erosion because both the coarse and fine contents runoff can be minimized while arranging of the slope angle is effective for reducing the runoff of coarse contents and the soil compaction is effective to reduce that of fine contents. Soil erosion can be minimized by selecting prevention method considering the type of soil because the prevention effect on soil erosion is different depending on the type of soil.

ADP-ribosylation factor-like 4c (ARL4C) is identified as a small GTP-binding protein, which is expressed by Wnt and EGF signaling and plays an important role in tubulogenesis of cultured cells and the ureters. ARL4C is little expressed in adult tissues, but it is highly expressed in lung cancer and colorectal cancer and shown to represent a molecular target for cancer therapy based on siRNA experiments. This study revealed that ARL4C is highly expressed in primary hepatocellular carcinoma (HCC) tumors and colorectal cancer liver metastases, and that ARL4C expression is associated with poor prognosis for these cancers. Chemically modified antisense oligonucleotides (ASO) against ARL4C effectively reduced ARL4C expression in both HCC and colorectal cancer cells and inhibited proliferation and migration of these cancer cells in vitro. ARL4C ASOs decreased the PIK3CD mRNA levels and inhibited the activity of AKT in HCC cells, suggesting that the downstream signaling of ARL4C in HCC cells is different from that in lung and colon cancer cells. In addition, subcutaneous injection of ARL4C ASO was effective in reducing the growth of primary HCC and metastatic colorectal cancer in the liver of immunodeficient mice. ARL4C ASO accumulated in cancer cells more efficiently than the surrounding normal cells in the liver and decreased ARL4C expression in the tumor. These results suggest that ARL4C ASO represents a novel targeted nucleic acid medicine for the treatment of primary and metastatic liver cancers.

Catchment water quality plays an important role in ecosystem and water resource management in mountainous areas. Shallow landslides triggered by earthquakes or heavy rainfall can cause a sudden and long-term deterioration in stream water quality by releasing contaminants into streams. Although many studies have been undertaken on the relationship between a single landslide and the water chemistry of a nearby river, little is known about the impact of densely distributed shallow landslides on stream water chemistry at the catchment scale. To this end, this study determined the major ion concentrations and isotopic compositions of stream water along with the shallow landslide area/catchment area ratio (LCR) in 37 headwater subcatchments in the southern part of Hokkaido, Japan, where an earthquake caused more than 6000 shallow landslides on September 6, 2018. In subcatchments with a high LCR, stream water exhibited significantly higher Ca