Syntenic analysis of ACCase loci and target-site-resistance mutations in cyhalofop-butyl resistant Echinochloa crus-galli var. crus-galli in Japan

BACKGROUND: Recently, suspected cyhalofop-butyl-resistant populations of allohexaploid weed Echinochloa crus-galli var. crus-galli were discovered in rice fields in Aichi Prefecture, Japan. Analyzing the target-site ACCase genes of cyhalofop-butyl helps understand the resistance mechanism. However, in E. crus-galli, the presence of multiple ACCase genes and the lack of detailed gene investigations have complicated the analysis of target-site genes. Therefore, in this study, we characterized the herbicide response of E. crus-galli lines and thoroughly characterized the ACCase genes, including the evaluation of gene mutations in the ACCase genes of each line. RESULT: Four suspected resistant lines collected from Aichi Prefecture showed varying degrees of resistance to cyhalofop-butyl and other FOP-class ACCase inhibitors but were sensitive to herbicides with other modes of action. Through genomic analysis, six ACCase loci were identified in the E. crus-galli genome. We renamed each gene based on its syntenic relationship with other ACCase genes in the Poaceae species. RNA-sequencing analysis revealed that all ACCase genes, except the pseudogenized copy ACCase2A, were transcribed at a similar level in the shoots of E. crus-galli. Mutations known to confer resistance to FOP-class herbicides, that is W1999C, W2027C/S and I2041N, were found in all resistant lines in either ACCase1A, ACCase1B or ACCase2C. CONCLUSION: In this study, we found that the E. crus-galli lines were resistant exclusively to ACCase-inhibiting herbicides, with a target-site resistance mutation in the ACCase gene. Characterization of ACCase loci in E. crus-galli provides a basis for further research on ACCase herbicide resistance in Echinochloa spp. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry

Questionnaire Survey on the Status of Promotion of Gender Equality and Diversity in Research : Current Situation at Tokushima University Based on Comparison with Nationwide Aggregated Results

本調査では，全国ダイバーシティネットワークの幹事機関である大阪大学と，日本学術会議科学者委員会男女共同参画分科会・同アンケート検討小分科会が共同で実施した「大学・研究機関における男女共同参画・ダイバーシティの推進状況調査」のうち「大学・研究機関における男女共同参画の推進状況に対する意見・感想」について，徳島大学回答分（男性60 件，女性21 件）のみ抽出されたデータを用いて，本学における男女共同参画・ダイバーシティ推進に関する研究者の意識調査を実施した。男女別に集計し，公開されている全国集計結果（計10，105 件）と比較検討することで，全国における本学の位置付けについて分析した。 　調査項目は， 1 ．男女共同参画の推進（全般） 2 ．男女共同参画の取り組み　3 ．ワークライフバランス・育児支援サービス　4 ．ダイバーシティ対応　5 ．ハラスメント防止体制　6 ．医学系　の6 項目にわたる。全国と徳島での回答差，また男女間の回答差から，現在の徳島大学の研究者における男女共同参画に対する意識について考察する。In this study, a survey was conducted to examine the attitude of researchers toward the promotion of gender equality and diversity at Tokushima University using the responses of only those from the university (60 male and 21 female), extracted from the survey segment “Opinions and Impressions on the Status of the Promotion of Gender Equality in Universities and Research Institutes.” The “Survey on the Promotion of Gender Equality and Diversity in Universities and Research Institutes” was conducted jointly by Osaka University, which is the secretariat of the National Diversity Network, and the Subcommittee on Gender Equality and the Committee for Scientific Community, the Science Council of Japan. By aggregating by gender and comparing the results with those published nationwide (total: 10,105 cases), this study analyzed the positioning of our university within Japan. The survey items were as follows: 1. promotion of gender equality (overall); 2. efforts for gender equality; 3. work-life balance and childcare support services; 4. support for diversity; 5. system to prevent harassment; and 6. the medical system. The differences in the responses between Japan and Tokushima as well as those between the male and female respondents were used to examine the attitude toward gender equality among current researchers at Tokushima University

Development of a Reduction‐Responsive Amino Acid that Induces Peptide Bond Cleavage in Hypoxic Cells

Utilization of a hypoxia-responsive amino acid is indispensable in the preparation of hypoxic tumor-specific peptidyl prodrugs. Bioreduction of a nitro group is among the most attractive triggering reactions in the hypoxia-responsive prodrugs. In this paper, design and synthesis of a reduction-responsive amino acid that induces peptide bond cleavage after reduction of the nitro group are described. Application to hypoxia-responsive peptide bond cleavage system is also reported

Development of a Reduction‐Responsive Amino Acid that Induces Peptide Bond Cleavage in Hypoxic Cells

Utilization of a hypoxia-responsive amino acid is indispensable in the preparation of hypoxic tumor-specific peptidyl prodrugs. Bioreduction of a nitro group is among the most attractive triggering reactions in the hypoxia-responsive prodrugs. In this paper, design and synthesis of a reduction-responsive amino acid that induces peptide bond cleavage after reduction of the nitro group are described. Application to hypoxia-responsive peptide bond cleavage system is also reported

CA9 and PRELID2; hypoxia-responsive potential therapeutic targets for pancreatic ductal adenocarcinoma as per bioinformatics analyses

A strong hypoxic environment has been observed in pancreatic ductal adenocarcinoma (PDAC) cells, which contributes to drug resistance, tumor progression, and metastasis. Therefore, we performed bioinformatics analyses to investigate potential targets for the treatment of PDAC. To identify potential genes as effective PDAC treatment targets, we selected all genes whose expression level was related to worse overall survival (OS) in The Cancer Genome Atlas (TCGA) database and selected only the genes that matched with the genes upregulated due to hypoxia in pancreatic cancer cells in the dataset obtained from the Gene Expression Omnibus (GEO) database. Although the extracted 107 hypoxia-responsive genes included the genes that were slightly enriched in angiogenic factors, TCGA data analysis revealed that the expression level of endothelial cell (EC) markers did not affect OS. Finally, we selected CA9 and PRELID2 as potential targets for PDAC treatment and elucidated that a CA9 inhibitor, U-104, suppressed pancreatic cancer cell growth more effectively than 5-fluorouracil (5-FU) and PRELID2 siRNA treatment suppressed the cell growth stronger than CA9 siRNA treatment. Thus, we elucidated that specific inhibition of PRELID2 as well as CA9, extracted via exhaustive bioinformatic analyses of clinical datasets, could be a more effective strategy for PDAC treatment

A Rat-Based Preclinical Platform Facilitating Transcatheter Hepatic Arterial Infusion in Immunodeficient Rats With Liver Xenografts of Patient-Derived Pancreatic Ductal Adenocarcinoma

Liver metastases from pancreatic ductal adenocarcinoma (PDAC) are highly fatal. A rat-based patient-derived tumor xenograft (PDX) model is available for transcatheter therapy. This study aimed to create an immunodeficient rat model with liver xenografts of patient-derived primary PDAC and evaluate efficacy of hepatic arterial infusion chemotherapy with cisplatin in this model. Three patient-derived PDACs were transplanted into the livers of 21 rats each (totally, 63 rats), randomly assigned into hepatic arterial infusion, systemic venous infusion, and control groups (n = 7 each) four weeks post-implantation. Computed tomography evaluated tumor volumes before and four weeks after treatment. Post-euthanasia, resected tumor specimens underwent histopathological examination. A liver-implanted PDAC PDX rat model was established in all 63 rats, with first CT identifying all tumors. Four weeks post-treatment, arterial infusion groups exhibited significantly smaller tumor volumes than controls for all three tumors on second CT. Xenograft tumors histologically maintained adenocarcinoma features compared to original patient tumors. Ki67 expression was significantly lower in arterial infusion groups than in the other two for the three tumors, indicating reduced tumor growth in PDX rats. A liver-implanted PDAC PDX rat model was established as a rat-based preclinical platform. Arterial cisplatin infusion chemotherapy represents a potential therapy for PDAC liver metastasis

Functional analyses of a novel missense and other mutations of the vitamin D receptor in association with alopecia

WOS: 000405172600069PubMed ID: 28698609Hereditary 1,25-dihydroxyvitamin D-resistant rickets (HVDRR) is a rare disorder, caused by bialellic mutations of the vitamin D receptor (VDR) gene, sometimes associated with alopecia. The aim of this study is to elucidate the mechanism of functional disruption of a novel mutation, detected in a patient with HVDRR, comparing to other mutations with or without alopecia. The patient was a 2-year-old girl with alopecia, who was clinically diagnosed as HVDRR. Genetic analysis revealed a novel homozygous mutation, S360P, located in ligand binding domain (LBD). The mutation was predicted as not disease causing by Polyphen2 and SIFT. But the transcriptional activity of S360P was disrupted as well as other reported mutations, Q152X (located in the hinge lesion), and R274L, H305Q (located in LBD). Following assays revealed no ligand binding affinity, no interaction with cofactors or RXR and no functioning of nuclear localization signals. Our results provide an additional evidence for the previous findings suggesting that DNA binding by the VDR/RXR heterodimer is essential for the function of the VDR in hair development. In conclusion, we identified a novel missense mutation of VDR causing HVDRR with alopecia. Functional analyses revealed that the single amino acid substitution could disrupt the function of the protein.Ministry of Education, Culture, Sports, Science, and Technology JapanMinistry of Education, Culture, Sports, Science and Technology, Japan (MEXT) [24791042, 23591489]; JSPS KAKENHIMinistry of Education, Culture, Sports, Science and Technology, Japan (MEXT)Japan Society for the Promotion of ScienceGrants-in-Aid for Scientific Research (KAKENHI) [JP16J04675]We are grateful to the patient and her parents for all their help and participation in this study. We thank Ms. Reiko Onai for technical support. This study was supported by a Grant-in-Aid from the Ministry of Education, Culture, Sports, Science, and Technology Japan (to TI: 24791042 and SK: 23591489), JSPS KAKENHI (to MT: JP16J04675)

Chloride Selective Macrocyclic Bisurea Derivatives with 2,2′‑Binaphthalene Moieties as Spacers

A cyclic bisurea derivative <b>2a</b> has been successfully prepared from the corresponding diamine and diisocyanate in the presence of tetrabutylammonium chloride as a template. A more soluble cyclic bisurea <b>2b</b> has also been prepared by introduction of sterically bulky <i>tert</i>-butyl groups. X-ray crystal analyses of [<b>2a</b>·Cl]⁻ and [<b>2b</b>·Cl]⁻ revealed that overall structure was saddle like and the chloride anion was located in the center of the cavity. The bound chloride anion was hydrogen bonded by four N–H of urea groups and weakly hydrogen bonded by four 1-C–H of naphthyl groups, respectively. After removal of the bound chloride anions of [<b>2b</b>·Cl]⁻ with silver nitrate, two different X-ray crystals of free <b>2b</b> were obtained; one was intermolecular hydrogen bonded shrunken structure and the other was extended structure. Receptor <b>2b</b> showed large binding ability for Cl^–, however, the selectivity for Cl^– against basic anions, such as AcO^– and F^–, has been insufficient. In aqueous MeCN, the association constant of <b>2b</b> for Cl^– was reduced but still large, and the selectivity for hydrophobic Cl^– was greatly improved. In this solvent, <b>2b</b> also selectively recognized alkaline metal chloride salts. Therefore, cyclic bisurea <b>2b</b> is highly selective and effective Cl^– selective receptor

Chloride Selective Macrocyclic Bisurea Derivatives with 2,2′‑Binaphthalene Moieties as Spacers

A cyclic bisurea derivative <b>2a</b> has been successfully prepared from the corresponding diamine and diisocyanate in the presence of tetrabutylammonium chloride as a template. A more soluble cyclic bisurea <b>2b</b> has also been prepared by introduction of sterically bulky <i>tert</i>-butyl groups. X-ray crystal analyses of [<b>2a</b>·Cl]⁻ and [<b>2b</b>·Cl]⁻ revealed that overall structure was saddle like and the chloride anion was located in the center of the cavity. The bound chloride anion was hydrogen bonded by four N–H of urea groups and weakly hydrogen bonded by four 1-C–H of naphthyl groups, respectively. After removal of the bound chloride anions of [<b>2b</b>·Cl]⁻ with silver nitrate, two different X-ray crystals of free <b>2b</b> were obtained; one was intermolecular hydrogen bonded shrunken structure and the other was extended structure. Receptor <b>2b</b> showed large binding ability for Cl^–, however, the selectivity for Cl^– against basic anions, such as AcO^– and F^–, has been insufficient. In aqueous MeCN, the association constant of <b>2b</b> for Cl^– was reduced but still large, and the selectivity for hydrophobic Cl^– was greatly improved. In this solvent, <b>2b</b> also selectively recognized alkaline metal chloride salts. Therefore, cyclic bisurea <b>2b</b> is highly selective and effective Cl^– selective receptor