72 results on '"Keita Yoshida"'
Search Results
2. Systemic amyloid A amyloidosis of the bladder after transurethral resection of urothelial carcinoma
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Kaori Yamashita, Keita Yoshida, Tadao Nakazawa, Satoshi Kubota, Takahiro Shiseki, Eri Sekido, and Masashi Inui
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amyloidosis ,bladder ,carcinoma ,Diseases of the genitourinary system. Urology ,RC870-923 - Abstract
Introduction Amyloid A amyloidosis of the bladder is not a major disease. We report a patient with systemic amyloid A amyloidosis of the bladder after transurethral resection of urothelial carcinoma. Case presentation An 87‐year‐old Japanese man had bladder carcinoma. He was followed up regularly with cystoscopy. Cystoscopy revealed multiple polypoid tumors 6 months after the first transurethral resection of urothelial carcinoma. Pathologic specimens contained the amyloid A component. He had hypertrophic cardiomyopathy, valvular disorders, and arrhythmias. His cardiac disease may have resulted from amyloid A amyloidosis. We speculated the patient had systemic amyloid A amyloidosis of the heart and bladder. Conclusion We determined the type of amyloidosis via a biopsy of the bladder tumors. Our patient had cardiac disease. Therefore, systemic amyloid A amyloidosis could have caused his cardiac disease. The pathologic findings of bladder tumors can contribute to detecting systemic amyloid A amyloidosis.
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- 2024
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3. Localized Amyloid A amyloidosis of ureter: A case report
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Kaori Yamashita, Keita Yoshida, Tadao Nakazawa, Takahiro Shiseki, Eri Sekido, Satoshi Kubota, and Masashi Inui
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Diseases of the genitourinary system. Urology ,RC870-923 - Abstract
Amyloidosis of the ureter is a rare disease, distinguishing it from a neoplasm is difficult. A 64-year-old Japanese woman suffered from macrohematuria and left side hydronephrosis with ultrasound in 2014. Retrograde pyelography revealed no ureter tumor at that time. The patient had macrohematuria, left side hydronephrosis, and ureteral stenosis in the left ureter on retrograde pyelography. She was suspected of having a ureter tumor when she was 71 years old in 2021. The patient underwent ureteroscopy with biopsy. Pathological specimens contained the amyloid A component, based on immunohistochemistry staining. We diagnosed the patient with amyloid A amyloidosis of the ureter.
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- 2023
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4. Distinct pathways for export of silencing RNA in Caenorhabditis elegans systemic RNAi
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Keita Yoshida, Yuji Suehiro, Katsufumi Dejima, Sawako Yoshina, and Shohei Mitani
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Natural sciences ,Biological sciences ,Biochemistry ,Genetics ,Molecular biology ,Cell biology ,Science - Abstract
Summary: Dietary supplied double-stranded RNA (dsRNA) can trigger RNA interference (RNAi) systemically in some animals, including the nematode Caenorhabditis elegans. Although this phenomenon has been utilized as a major tool for gene silencing in C. elegans, how cells spread the silencing RNA throughout the organism is largely unknown. Here, we identify two novel systemic RNAi-related factors, REXD-1 and TBC-3, and show that these two factors together with SID-5 act redundantly to promote systemic spreading of dsRNA. Animals that are defective in all REXD-1, TBC-3, and SID-5 functions show strong deficiency in export of dsRNA from intestinal cells, whereas cellular uptake and processing of dsRNA and general secretion events other than dsRNA secretion are still functional in the triple mutant animals. Our findings reveal pathways that specifically regulate the export of dsRNA in parallel, implying the importance of spreading RNA molecules for intercellular communication in organisms.
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- 2023
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5. An endomembrane zinc transporter negatively regulates systemic RNAi in Caenorhabditis elegans
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Katsufumi Dejima, Rieko Imae, Yuji Suehiro, Keita Yoshida, and Shohei Mitani
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Molecular biology ,Cell biology ,Science - Abstract
Summary: Double-stranded RNA (dsRNA) regulates gene expression in a sequence-dependent manner. In Caenorhabditis elegans, dsRNA spreads through the body and leads to systemic RNA silencing. Although several genes involved in systemic RNAi have been genetically identified, molecules that mediate systemic RNAi remain largely unknown. Here, we identified ZIPT-9, a C. elegans homolog of ZIP9/SLC39A9, as a broad-spectrum negative regulator of systemic RNAi. We showed that RSD-3, SID-3, and SID-5 genetically act in parallel for efficient RNAi, and that zipt-9 mutants suppress the RNAi defects of all the mutants. Analysis of a complete set of deletion mutants for SLC30 and SLC39 family genes revealed that only zipt-9 mutants showed altered RNAi activity. Based on these results and our analysis using transgenic Zn2+ reporters, we propose that ZIPT-9-dependent Zn2+ homeostasis, rather than overall cytosolic Zn2+, modulates systemic RNAi activity. Our findings reveal a previously unknown function of zinc transporters in negative RNAi regulation.
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- 2023
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6. Expression of T-cell immunoreceptor with immunoglobulin and tyrosine-based inhibitory motif domains (TIGIT) in anaplastic thyroid carcinoma
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Tadao Nakazawa, Takuya Nagasaka, Keita Yoshida, Atsuko Hasegawa, Feng Guo, Di Wu, Kenzo Hiroshima, and Ryohei Katoh
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TIGIT ,Anaplastic thyroid carcinoma ,Poorly differentiated thyroid carcinoma ,Medullary thyroid carcinoma ,Immunohistochemistry ,Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Abstract
Abstract Background Immune checkpoint proteins have not been fully examined in follicular cell-derived thyroid carcinoma and medullary thyroid carcinoma (MTC). Anaplastic thyroid carcinoma (ATC) is one of the most aggressive carcinomas. Even multimodal treatment does not result in favorable clinical outcomes for patients with ATC. Anti-tumor immunity has therefore been highlighted as having therapeutic promise for ATC. Methods We examined a novel immune checkpoint receptor, T-cell immunoreceptor with immunoglobulin and tyrosine-based inhibitory motif domains (TIGIT), in variable thyroid lesions: adenomatous goiter, follicular adenoma, and thyroid carcinoma (TC) using immunohistochemistry (IHC). Results Our IHC results showed that TIGIT expression was detected in cancer cells of MTC and high-grade TC: poorly differentiated thyroid carcinoma (PDTC) and ATC. Neoplastic cells were positive for TIGIT in four of five MTCs (80.0%), 17 of 31 ATCs (54.8%) and in 3 of 12 PDTCs (25.0%). TIGIT was not detected in any adenomatous goiters, thyroid benign tumors, or differentiated thyroid carcinoma (DTCs). Intriguingly, ATC cells showing pleomorphic/giant cell features were positive for TIGIT, while ATC cells with other cell morphologies lacked the immunoreactivity. Intra-tumoral immune cell was inclined to be enriched in TIGI-positive ATC. Although coexisting papillary thyroid carcinoma (PTC) components demonstrated high-grade microscopic features, neither the PTC nor follicular thyroid carcinoma (FTC) components expressed TIGT in any composite ATCs. Conclusion TIGIT was immunohistochemically found in MTC with high frequency and partially in high-grade TC. TIGIT expression in cancer cells may be beneficial for a potential utility in MTC and a subset of high-grade TC, especially ATC therapy.
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- 2022
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7. Association and dose-response relationship between exposure to alcohol advertising media and current drinking: a nationwide cross-sectional study of Japanese adolescents
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Keita Yoshida, Hideyuki Kanda, Takashi Hisamatsu, Yuki Kuwabara, Aya Kinjo, Hisashi Yoshimoto, Teruna Ito, Hideaki Kasuga, Ruriko Minobe, Hitoshi Maesato, Maki Jike, Yuuki Matsumoto, Yuichiro Otsuka, Osamu Itani, Yoshitaka Kaneita, Susumu Higuchi, and Yoneatsu Osaki
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underage drinking ,alcohol ,adolescents ,advertisement ,Public aspects of medicine ,RA1-1270 - Abstract
Background: Underage drinking is a public health concern. However, few studies have examined the association between alcoholic beverage advertising and underage drinking, particularly in countries with low underage drinking rates, such as Japan. Therefore, we aimed to investigate the relationship between exposure to advertising in various media and alcohol drinking among Japanese adolescents. Methods: We conducted a cross-sectional study involving 15,683 adolescents (51% girls) using data from a nationwide lifestyle survey in 2021 among junior and senior high schools across Japan. Media types were websites, stores, and public transportation. We defined current drinking as alcohol consumption of ≥1 day in the 30 days preceding the survey. Multivariable logistic regression was used to examine the association between exposure to alcohol advertisements and current drinking, adjusting for sex, grades, school area, lifestyle (bedtime and having fun at school), and addictive behaviors (smoking status and parents’ alcohol consumption). Results: The prevalence of current drinking was 2.2% (2.3% of boys and 2.0% of girls). Students who were exposed to any alcohol advertising media had higher odds of current drinking compared with those who were not (odds ratio, 1.48; 95% confidence interval [CI], 1.18–1.87). Students who were exposed to web, in-store, and public transportation advertisements had odds ratios of 1.44 (95% CI, 1.14–1.81), 1.62 (1.28–2.05), and 1.45 (1.06–1.98) of current drinking, respectively, compared with those who were not. The association of exposure to alcohol advertising media with the prevalence of current drinking was similar among boys and girls (all p for sex interaction >0.1), except for that of exposure to web advertisements; its association with current drinking was more pronounced in girls (p for sex interaction = 0.046). Exposure to a larger cumulative number of different alcohol advertising media was independently associated with a higher prevalence of current drinking among all students, boys, and girls (p-values for trend
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- 2023
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8. Application of detergents or high hydrostatic pressure as decellularization processes in uterine tissues and their subsequent effects on in vivo uterine regeneration in murine models.
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Erna G Santoso, Keita Yoshida, Yasushi Hirota, Masanori Aizawa, Osamu Yoshino, Akio Kishida, Yutaka Osuga, Shigeru Saito, Takashi Ushida, and Katsuko S Furukawa
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Medicine ,Science - Abstract
Infertility caused by ovarian or tubal problems can be treated using In Vitro Fertilization and Embryo Transfer (IVF-ET); however, this is not possible for women with uterine loss and malformations that require uterine reconstruction for the treatment of their infertility. In this study, we are the first to report the usefulness of decellularized matrices as a scaffold for uterine reconstruction. Uterine tissues were extracted from Sprague Dawley (SD) rats and decellularized using either sodium dodecyl sulfate (SDS) or high hydrostatic pressure (HHP) at optimized conditions. Histological staining and quantitative analysis showed that both SDS and HHP methods effectively removed cells from the tissues with, specifically, a significant reduction of DNA contents for HHP constructs. HHP constructs highly retained the collagen content, the main component of extracellular matrices in uterine tissue, compared to SDS constructs and had similar content levels of collagen to the native tissue. The mechanical strength of the HHP constructs was similar to that of the native tissue, while that of the SDS constructs was significantly elevated. Transmission electron microscopy (TEM) revealed no apparent denaturation of collagen fibers in the HHP constructs compared to the SDS constructs. Transplantation of the decellularized tissues into rat uteri revealed the successful regeneration of the uterine tissues with a 3-layer structure 30 days after the transplantation. Moreover, a lot of epithelial gland tissue and Ki67 positive cells were detected. Immunohistochemical analyses showed that the regenerated tissues have a normal response to ovarian hormone for pregnancy. The subsequent pregnancy test after 30 days transplantation revealed successful pregnancy for both the SDS and HHP groups. These findings indicate that the decellularized matrix from the uterine tissue can be a potential scaffold for uterine regeneration.
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- 2014
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9. Radiation Hardened Flip-Flops with low Area, Delay and Power Overheads in a 65 nm bulk process.
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Shotaro Sugitani, Ryuichi Nakajima, Keita Yoshida, Jun Furuta, and Kazutoshi Kobayashi
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- 2023
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10. SEU Sensitivity of PMOS and NMOS Transistors in a 65 nm Bulk Process by α-Particle Irradiation.
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Keita Yoshida, Ryuichi Nakajima, Shotaro Sugitani, Takafumi Ito, Jun Furuta, and Kazutoshi Kobayashi
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- 2023
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11. Prognostic Evaluation of HER2-positive Early Breast Cancers Using Clinico-pathological Criteria.
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NORIE JIBIKI, TADAO NAKAZAWA, MAKO NOGAMI, KEITA YOSHIDA, ATSUKO HASEGAWA, FENG GUO, and KENZO HIROSHIMA
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HER2 positive breast cancer ,NEOADJUVANT chemotherapy ,TUMOR classification ,OVERALL survival ,UNIVARIATE analysis - Abstract
Background/Aim: HER2-positive breast carcinomas (BCs) generally behave more aggressively and show higher cytological and histological grade than HER2-negative BCs. However, the clinical properties of HER2-positive early BCs have not been studied extensively. Hence, the therapeutic significance of neoadjuvant chemotherapy (NAC) for this BC remains debatable. Patients and Methods: We retrospectively examined the clinicopathological features of 94 HER2-positive early BCs who perioperatively received anti-HER2 drugs, without undergoing NAC prior to surgery. Results: The patients' five year-disease free survival (DFS) and overall survival (OS) rates were 95.6% and 100%, respectively. Univariate analysis demonstrated significant differences in distant metastasis-free survival (DMFS) between clinical and pathological tumor stages (T stages). Pathological T1 stage and clinical T1 stage tumors showed significantly higher DMSF than pT2-3 and cT2-3 (p=0.0002 and 0.0294). Multivariate analysis disclosed no significant differences in DFS, OS, and DMFS with respect to preoperative clinical tumor stage, patient age, type of surgery, postoperative therapy, and pathological factors. Recurrences occurred in nine patients: four (4.3%) and five (5.3%) patients showed local and distant recurrences, respectively. One patient with cT2 BC died of disease. Interestingly, four of the five BCs with distant recurrence pathologically demonstrated lymph vessel invasion. The prognoses of patients with HER2-positive stage cT1/2N0M0 BC were highly favorable. Conclusion: The indications for NAC in small, localized, and node-negative HER2-positive BC should be carefully assessed based on the presence of a larger tumor size, postoperative pathological evaluation of tumor size, and lymph vessel invasion. [ABSTRACT FROM AUTHOR]
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- 2024
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12. Oligodeoxynucleotides Modified with 2′-O-(Cysteinylaminobutyl)carbamoylethylribothymidine Residues for Native Chemical Ligation with Peptide at Internal Positions
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Yoshiaki Masaki, Atsuya Maruyama, Keita Yoshida, Takahito Tomori, Tomohiro Kishimura, and Kohji Seio
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Pharmacology ,Organic Chemistry ,Biomedical Engineering ,Pharmaceutical Science ,Bioengineering ,Biotechnology - Published
- 2022
13. Hybridization of an n-type semiconducting polymer with PbS quantum dots and their photovoltaic investigation
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Keita Yoshida, Jia-Fu Chang, Chu-Chen Chueh, and Tomoya Higashihara
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Polymers and Plastics ,Materials Chemistry - Published
- 2022
14. Improvement of (S)-selective imine reductase GF3546 for the synthesis of chiral cyclic amines
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Yuta Fukawa, Keita Yoshida, Sayaka Degura, Koichi Mitsukura, and Toyokazu Yoshida
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Materials Chemistry ,Metals and Alloys ,Ceramics and Composites ,General Chemistry ,Catalysis ,Surfaces, Coatings and Films ,Electronic, Optical and Magnetic Materials - Abstract
Imine reductase (IRED) from Streptomyces sp. GF3546, one of the first IREDs, was engineered and showed excellent performance at a high concentration of 2-methyl-1-pyrroline and high thermostability at 50 °C.
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- 2022
15. Strain-insensitive naphthalene-diimide-based conjugated polymers through sequential regularity control
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Yan-Cheng Lin, Kosuke Terayama, Keita Yoshida, Ping-Jui Yu, Pin-Hsiang Chueh, Chu-Chen Chueh, Tomoya Higashihara, and Wen-Chang Chen
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Materials Chemistry ,General Materials Science - Abstract
Sequential regularity control on the n-type conjugated polymers was investigated in this work. The sequentially random polymer produced a near-amorphous structure and a strain-insensitive charge transport performance.
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- 2022
16. Improvement of (
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Yuta, Fukawa, Keita, Yoshida, Sayaka, Degura, Koichi, Mitsukura, and Toyokazu, Yoshida
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Stereoisomerism ,Imines ,Amines ,Oxidoreductases ,Streptomyces ,Amination - Abstract
An engineered (
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- 2022
17. Thiol-end-functionalized Regioregular Poly(3-hexylthiophene) for PbS Quantum Dot Dispersions
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Keita Yoshida, Jia Fu Chang, Chu-Chen Chueh, and Tomoya Higashihara
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chemistry.chemical_classification ,Materials science ,Polymers and Plastics ,chemistry ,Quantum dot ,Process Chemistry and Technology ,Organic Chemistry ,Thiol ,Photochemistry - Published
- 2021
18. Hox13 is essential for formation of a sensory organ at the terminal end of the sperm duct in Ciona
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Yasunori Sasakura, Yukako Tajima, Nicholas Treen, Takashi Yamamoto, Keita Yoshida, Akiko Hozumi, and Tetsushi Sakuma
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Male ,animal structures ,Ciona savignyi ,Genitalia, Male ,Models, Biological ,03 medical and health sciences ,0302 clinical medicine ,Species Specificity ,Animals ,Ciona intestinalis ,Hox gene ,Molecular Biology ,Transcription factor ,Gene ,030304 developmental biology ,Neurons ,0303 health sciences ,biology ,fungi ,Genes, Homeobox ,Gene Expression Regulation, Developmental ,Sense Organs ,Epithelial Cells ,Pigments, Biological ,Cell Biology ,biology.organism_classification ,Sperm ,Cell biology ,Ciona ,Pseudopodia ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Species-specific traits are thought to have been acquired by natural selection. Transcription factors play central roles in the evolution of species-specific traits. Hox genes encode a set of conserved transcription factors essential for establishing the anterior-posterior body axis of animals. Changes in the expression or function of Hox genes can lead to the diversification of animal-body plans. The tunicate ascidian Ciona intestinalis Type A has an orange-colored structure at the sperm duct terminus. This orange-pigmented organ (OPO) is the characteristic that can distinguish this ascidian from other closely related species. The OPO is formed by the accumulation of orange-pigmented cells (OPCs) that are present throughout the adult body. We show that Hox13 is essential for formation of the OPO. Hox13 is expressed in the epithelium of the sperm duct and neurons surrounding the terminal openings for sperm ejection, while OPCs themselves do not express this gene. OPCs are mobile cells that can move through the body vasculature by pseudopodia, suggesting that the OPO is formed by the accumulation of OPCs guided by Hox13-positive cells. Another ascidian species, Ciona savignyi, does not have an OPO. Like Hox13 of C. intestinalis, Hox13 of C. savignyi is expressed at the terminus of its sperm duct; however, its expression domain is limited to the circular area around the openings. The genetic changes responsible for the acquisition or loss of OPO are likely to occur in the expression pattern of Hox13.
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- 2020
19. Synthesis of 2′-O-(N-methylcarbamoylethyl) 5-methyl-2-thiouridine and its application to splice-switching oligonucleotides
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Jun Tanihata, Keishi Yamamoto, Kohji Seio, Shin'ichi Takeda, Tetsuya Nagata, Keita Yoshida, Atsuya Maruyama, Takeshi Inde, Yoshiaki Masaki, and Mitsuo Sekine
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chemistry.chemical_classification ,5-methyl-2-thiouridine ,Molecular Structure ,010405 organic chemistry ,Oligonucleotide ,Stereochemistry ,Thiouridine ,Organic Chemistry ,Clinical Biochemistry ,Oligonucleotides ,Pharmaceutical Science ,01 natural sciences ,Biochemistry ,Exon skipping ,0104 chemical sciences ,010404 medicinal & biomolecular chemistry ,chemistry ,Splice switching ,Drug Discovery ,RNA splicing ,Molecular Medicine ,Nucleotide ,Molecular Biology - Abstract
The effect of 2'-O-(N-methylcarbamoyl)ethyl (MCE) modification on splice-switching oligonucleotides (SSO) was systematically evaluated. The incorporation of five MCE nucleotides at the 5'-termini of SSOs effectively improved the splice switching effect. In addition, the incorporation of 2'-O-(N-methylcarbamoylethyl)-5-methyl-2-thiouridine (s2TMCE), a duplex-stabilizing nucleotide with an MCE modification, into SSOs further improved splice switching. These SSOs may be useful for the treatment of genetic diseases associated with splicing errors.
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- 2019
20. Effective Postoperative Analgesia Using Intravenous Flurbiprofen and Acetaminophen
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Masahiro Irifune, Mitsuru Doi, Akari Mukai, Mitsuhiro Yoshida, Keita Yoshida, and Yoshitaka Shimizu
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Adult ,Male ,Drug ,medicine.medical_specialty ,media_common.quotation_subject ,Oral Surgical Procedures ,Flurbiprofen ,Analgesic ,digestive system ,Fentanyl ,03 medical and health sciences ,0302 clinical medicine ,030202 anesthesiology ,030225 pediatrics ,Humans ,Medicine ,Mandibular Diseases ,Acetaminophen ,Pain Measurement ,media_common ,Pain, Postoperative ,Cysts ,business.industry ,Anti-Inflammatory Agents, Non-Steroidal ,Analgesics, Non-Narcotic ,Middle Aged ,digestive system diseases ,Discontinuation ,Analgesics, Opioid ,stomatognathic diseases ,Treatment Outcome ,Otorhinolaryngology ,Anesthesia ,Dental surgery ,Anesthetic ,Administration, Intravenous ,Drug Therapy, Combination ,Surgery ,Oral Surgery ,business ,medicine.drug - Abstract
Management of postoperative pain is one of the most important components in postoperative care, because most patients have pain after dental surgery. The aim of this study was to evaluate whether acetaminophen could be an alternative to fentanyl in combination with a nonsteroidal anti-inflammatory drug (NSAID) as an analgesic after dental surgery in cases in which narcotic drugs were contraindicated.Patients were 24- to 54-year-old men who underwent enucleation of a mandibular cyst under general anesthesia. The authors measured time from discontinuation of anesthetic administration until discharge from the operating room and postoperative pain during 4 hours after discharge. They compared these parameters between patients who were intravenously administered an NSAID such as flurbiprofen with fentanyl (NSAID/fentanyl group) and those administered an NSAID with acetaminophen (NSAID/acetaminophen group). Parametric data of time were analyzed using Student t test. Nonparametric data of the analgesic effect were analyzed using Mann-Whitney U test.Time until discharge from the operating room after discontinuation of anesthetics in the NSAID/fentanyl group was significantly longer than that in the NSAID/acetaminophen group (P .05). In contrast, there was no significant difference in analgesic effect between the NSAID/acetaminophen and NSAID/fentanyl groups (P .05).Although recovery time in the operating room of the flurbiprofen and acetaminophen group was markedly shorter than that of the flurbiprofen and fentanyl group, the postoperative analgesic effects of the 2 drugs were equipotent. Therefore, acetaminophen can be an alternative to fentanyl in cases in which narcotic drugs are contraindicated.
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- 2018
21. Treatment of dental caries in a patient with Joubert syndrome without the use of sedatives: A case study
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Chiaki Furutani, Atsuko Wakita, Miwa Miyauchi, Keita Yoshida, Yoshiyuki Okada, Yuki Oda, and Yoko Hama
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Pediatrics ,medicine.medical_specialty ,Ataxia ,business.industry ,Apnea ,030206 dentistry ,Gene mutation ,medicine.disease ,Oral hygiene ,Joubert syndrome ,Cerebral palsy ,stomatognathic diseases ,03 medical and health sciences ,Epilepsy ,0302 clinical medicine ,stomatognathic system ,Intellectual disability ,medicine ,030212 general & internal medicine ,medicine.symptom ,business ,General Dentistry - Abstract
Background Joubert syndrome (JS) is a rare autosomal recessive inherited ciliopathy caused by gene mutation. Manifestations can include intermittent dyspnea, apnea, ataxia, and other nervous system abnormalities. Case presentation The patient was a 21-year-old female with JS, severe intellectual disability, cerebral palsy, and epilepsy. Dental caries in both mandibular lower first molars was diagnosed by a local dentist, and the patient was referred to us for further treatment. Although her oral hygiene was good, the dental caries had reached the vicinity of the dental pulp. The caries had developed symmetrically on both first molars and was completely covered with the gingiva. As she was hesitant to receive treatment without anxiolysis, we considered using general anesthesia or intravenous sedation. However, we were recommended against it by the patient's family doctor because of the risk of apneic episodes in JS. Accordingly, dental treatment was performed over multiple days through physical behavior adjustment, while providing oxygenation (3 L/min) in preparation for an apneic episode. The dental treatment was successfully completed with a good postoperative outcome. Conclusions Dental treatment can be safely performed in patients with JS through physical behavior adjustment, thereby minimizing the risk of an apnea attack.
- Published
- 2021
22. Modification of oligonucleotides with weak basic residues via the 2'-O-carbamoylethyl linker for improving nuclease resistance without loss of duplex stability and antisense activity
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Takahito Tomori, Atsuya Maruyama, Hiroyuki Nakajima, Yusuke Iriyama, Keishi Yamamoto, Keita Yoshida, Yoshiaki Masaki, Tatsuro Kanaki, and Kohji Seio
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Exonuclease ,0303 health sciences ,Nuclease ,biology ,010405 organic chemistry ,Chemistry ,RNase P ,Oligonucleotide ,Organic Chemistry ,Oligonucleotides ,01 natural sciences ,Biochemistry ,Combinatorial chemistry ,0104 chemical sciences ,03 medical and health sciences ,Mice ,Ribonucleases ,Duplex (building) ,Nucleic acid ,biology.protein ,Animals ,Nucleic Acid Conformation ,Physical and Theoretical Chemistry ,Linker ,030304 developmental biology - Abstract
For the improvement of nuclease resistance, four kinds of new modifications through a carbamoylethyl linker were designed. Among them, the 2′-O-[2-N-{2-(benzimidazol-1-yl)ethyl}carbamoylethyl] modification showed 20-fold longer half-life when treated with a 3′ to 5′ exonuclease compared to the 2′-O-methoxyethyl (MOE) modification, which is used in approved drugs. In addition, this large modification did not disturb the binding affinity or RNase H-dependent antisense activity. From these findings, it could be concluded that an adequate linker, such as carbamoylethyl in this study, could extend the utility of 2′-O-modification without loss of the properties of nucleic acids. This strategy would be useful for the development of nucleic acid therapeutics.
- Published
- 2019
23. TALEN-Based Knockout System
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Keita Yoshida and Nicholas Treen
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0301 basic medicine ,Transcription activator-like effector nuclease ,Cas9 ,fungi ,Mutagenesis (molecular biology technique) ,Computational biology ,Biology ,biology.organism_classification ,03 medical and health sciences ,030104 developmental biology ,Genome editing ,embryonic structures ,Conditional gene knockout ,Gene Knockout Techniques ,CRISPR ,Ciona intestinalis - Abstract
Targeted mutagenesis of genes-of-interest is a powerful method of addressing the functions of genes. Genome editing techniques, such as transcriptional activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 systems, have enabled this approach in various organisms because of their ease of use. In the ascidian, Ciona intestinalis, recent studies show that TALEN-based knockout can be applied to establishing both mutant lines and tissue-specific knockout for addressing gene functions. Here, we introduce recent updates to the TALEN toolkit that facilitate detailed functional analysis of genes in ascidians.
- Published
- 2018
24. Chondroitin 6-O-sulfotransferases are required for morphogenesis of the notochord in the ascidian embryo
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Yasunori Sasakura, Keita Yoshida, Shigeki Fujiwara, and Jun Nakamura
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Genetics ,animal structures ,biology ,Convergent extension ,fungi ,Morphogenesis ,Neural tube ,Embryo ,biology.organism_classification ,Cell biology ,chemistry.chemical_compound ,medicine.anatomical_structure ,chemistry ,embryonic structures ,Notochord ,medicine ,Chondroitin ,Ciona intestinalis ,Chondroitin sulfate ,Developmental Biology - Abstract
Background: Chondroitin sulfate (CS) is a sulfated polysaccharide chain that binds to various core proteins to form proteoglycans. The amount and position of sulfate groups in CS are variable among different tissues, and are determined by specific sulfotransferases. Although the ascidians are the closest relatives of vertebrates, the functions of their sulfotransferases have not been studied. Results: The genome of the ascidian Ciona intestinalis contains eight genes encoding proteins similar to chondroitin 6-O-sulfotransferases (C6STs), which appear to have independently diverged in the ascidian lineage during evolution. Among them, Ci-C6ST-like1 and Ci-C6ST-like7 were predominantly expressed in the developing notochord. In addition, they were weakly expressed in the neural tube. The disruption of either one of them affected the convergent extension movement of notochordal cells. Presumptive notochord cells coming from both sides of the embryo did not intercalate. The results suggest that both of them are necessary. In some cases, the anterior neural tube failed to close. Forced expression of Ci-C6ST-like1 or Ci-C6ST-like7 in the notochord restored the normal intercalation of notochordal cells, indicating that the effects of morpholino oligos are specific. Conclusions: Ci-C6ST-like1 and Ci-C6ST-like7 are required for the morphogenesis of the notochord in the ascidian embryo. Developmental Dynamics 243:1637–1645, 2014. © 2014 Wiley Periodicals, Inc.
- Published
- 2014
25. CRISPR/Cas9-mediated gene knockout in the ascidian Ciona intestinalis
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Akiko Hozumi, Keita Yoshida, Yasunori Sasakura, and Haruka Sasaki
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animal structures ,ascidian ,Genetic Vectors ,Mutagenesis (molecular biology technique) ,knockout ,Biology ,Polymerase Chain Reaction ,Gene Knockout Techniques ,Genome editing ,Mutation Rate ,CRISPR ,Animals ,Ciona intestinalis ,CRISPR/Cas9 ,Gene knockout ,DNA Primers ,Genetics ,CRISPR interference ,Cas9 ,fungi ,Genes, Homeobox ,Cell Biology ,Original Articles ,Genomics ,biology.organism_classification ,Ciona ,Electroporation ,Mutagenesis ,embryonic structures ,CRISPR-Cas Systems ,Developmental Biology - Abstract
Knockout of genes with CRISPR/Cas9 is a newly emerged approach to investigate functions of genes in various organisms. We demonstrate that CRISPR/Cas9 can mutate endogenous genes of the ascidian Ciona intestinalis, a splendid model for elucidating molecular mechanisms for constructing the chordate body plan. Short guide RNA (sgRNA) and Cas9 mRNA, when they are expressed in Ciona embryos by means of microinjection or electroporation of their expression vectors, introduced mutations in the target genes. The specificity of target choice by sgRNA is relatively high compared to the reports from some other organisms, and a single nucleotide mutation at the sgRNA dramatically reduced mutation efficiency at the on-target site. CRISPR/Cas9-mediated mutagenesis will be a powerful method to study gene functions in Ciona along with another genome editing approach using TALE nucleases.
- Published
- 2014
26. Genome Editing of the Ascidian Ciona intestinalis with TALE Nuclease
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Yasunori, Sasakura, Keita, Yoshida, and Nicholas, Treen
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Gene Editing ,Gene Knockout Techniques ,Electroporation ,Organ Specificity ,Transcription Activator-Like Effector Nucleases ,Animals ,Ciona intestinalis - Abstract
The ascidian Ciona intestinalis is an important model animal for studying developmental mechanisms for constructing the chordate body. Although molecular and embryological techniques for manipulating Ciona genes were developed a long time ago, recent achievements of genome editing in this animal have innovated functional analyses of genes in Ciona. Particularly, knockout of genes in the G0 generation coupled with tissue-specific expression of TALENs enables us to rapidly address gene functions that were difficult using previous methods.
- Published
- 2017
27. Hox-mediated endodermal identity patterns the pharyngeal muscle formation in the chordate pharynx
- Author
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Azusa Nakahata, Takashi Yamamoto, Keita Yoshida, Nicholas Treen, Yasunori Sasakura, and Tetsushi Sakuma
- Subjects
0301 basic medicine ,Mesoderm ,animal structures ,Tretinoin ,Chordate ,Muscle Development ,Pharyngeal muscles ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Ciona intestinalis ,Hox gene ,Molecular Biology ,Feedback, Physiological ,Homeodomain Proteins ,biology ,Endoderm ,Anatomy ,biology.organism_classification ,Ciona ,030104 developmental biology ,medicine.anatomical_structure ,embryonic structures ,Pharyngeal Muscles ,Pharynx ,030217 neurology & neurosurgery ,Developmental Biology ,Endostyle - Abstract
The pharynx, possessing gill slits and the endostyle, is a characteristic of chordates that is a complex of multiple tissues well organized along the anterior-posterior (AP) axis. Although Hox genes show AP coordinated expression in the pharyngeal endoderm, tissue specific roles of these factors for establishing the regional identities within this tissue is largely unknown. Here, we show that Hox1 is essential for the establishment of AP axial identity of the endostyle, a major structure of the pharyngeal endoderm, in the ascidian Ciona intestinalis. We found that Hox1 knockout causes posterior to anterior transformation of the endostyle identity, and Hox1 represses Otx expression and anterior identity, and vice versa. Furthermore, alteration of the regional identity of the endostyle disrupts the formation of body wall muscles, suggesting that the endodermal axial identity is essential for the coordinated pharyngeal development. Our results reveal an essential role of Hox genes for establishment of the AP regional identity in the pharyngeal endoderm and crosstalk between endoderm and mesoderm for the development of chordate pharynx.
- Published
- 2017
28. Nonreproductive role of gonadotropin-releasing hormone in the control of ascidian metamorphosis
- Author
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Honoo Satake, Chisato Kamiya, Naoyuki Ohta, Takeo Horie, Takehiro Kusakabe, Yasunori Sasakura, Keita Yoshida, and Yosuke Ogura
- Subjects
Larva ,medicine.medical_specialty ,animal structures ,biology ,media_common.quotation_subject ,fungi ,Gonadotropin-releasing hormone ,biology.organism_classification ,Tunicate ,Ciona ,Endocrinology ,Internal medicine ,Second messenger system ,medicine ,Ciona intestinalis ,Metamorphosis ,Developmental Biology ,media_common ,Hormone - Abstract
Background: Gonadotropin-releasing hormones (GnRHs) are neuropeptides that play central roles in the reproduction of vertebrates. In the ascidian Ciona intestinalis, GnRHs and their receptors are expressed in the nervous systems at the larval stage, when animals are not yet capable of reproduction, suggesting that the hormones have non-reproductive roles. Results: We showed that GnRHs in Ciona are involved in the animal 0 s metamorphosis by regulating tail absorption and adult organ growth. Absorption of the larval tail and growth of the adult organs are two major events in the metamorphosis of ascidians. When larvae were treated with GnRHs, they completed tail absorption more frequently than control larvae. cAMP was suggested to be a second messenger for the induction of tail absorption by GnRHs. tGnRH-3 and tGnRH-5 (the “t” indicates “tunicate”) inhibited the growth of adult organs by arresting cell cycle progression in parallel with the promotion of tail absorption. Conclusions: This study provides new insights into the molecular mechanisms of ascidian metamorphosis conducted by non-reproductive GnRHs. Developmental Dynamics 00:000–000, 2014. V C 2014 Wiley Periodicals, Inc.
- Published
- 2014
29. Germ cell mutations of the ascidianCiona intestinaliswith TALE nucleases
- Author
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Nicholas Treen, Keita Yoshida, Yasunori Sasakura, Takashi Yamamoto, Akiko Hozumi, and Tetsushi Sakuma
- Subjects
Genetics ,Transcription activator-like effector nuclease ,biology ,Somatic cell ,fungi ,Mutagenesis (molecular biology technique) ,Cell Biology ,biology.organism_classification ,Genome ,Ciona ,Endocrinology ,medicine.anatomical_structure ,embryonic structures ,medicine ,Ciona intestinalis ,Gene ,Germ cell - Abstract
Summary: Targeted mutagenesis of genes-of-interest, or gene-knockout, is a powerful method to address the functions of genes. Engineered nucleases have enabled this approach in various organisms because of their ease of use. The ascidian Ciona intestinalis is an excellent organism to analyze gene functions by means of genetic technologies. In our previous study, we reported mutagenesis of Ciona somatic cells with TALE nucleases (TALENs) by electroporating expression constructs. In this study, we report germ cell mutagenesis of Ciona by microinjecting mRNAs encoding TALENs. TALEN mRNAs introduced mutations to target genes in both somatic and germ cells. TALEN-mediated mutations in the germ cell genome were inherited by the next generation. We conclude that knockout lines of Ciona that have disrupted target genes can be established through TALEN-mediated germ cell mutagenesis. genesis 52:431–439, 2014. © 2014 Wiley Periodicals, Inc.
- Published
- 2014
30. Thermal conductivities of ThO2, NpO2 and their related oxides: Molecular dynamics study
- Author
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Taku Matsumoto, Tatsumi Arima, Keita Yoshida, Kazuya Idemitsu, and Yaohiro Inagaki
- Subjects
Nuclear and High Energy Physics ,Phonon scattering ,Chemistry ,Ionic bonding ,Thermodynamics ,Actinide ,Lattice strain ,Molecular dynamics ,Thermal conductivity ,Nuclear Energy and Engineering ,Thermal ,Physical chemistry ,Condensed Matter::Strongly Correlated Electrons ,General Materials Science ,Solid solution - Abstract
The thermal conductivities of ThO 2 , NpO 2 , (Th, U)O 2 , (Th, Pu)O 2 and (U, Np)O 2 have been investigated by molecular dynamics (MD) simulation up to 2000 K using the Busing–Ida potential function with partial ionic charges. In the present study, the thermal conductivity was calculated mainly by the Green–Kubo formula in the equilibrium MD scheme. The thermal conductivities of above actinide dioxides decreased with the increase of temperature due to the phonon–phonon interaction (Umklapp process). Concerning the composition of solid solutions, the decrease in thermal conductivity of (Th, Pu)O 2 is great as compared to other ones. Various MD calculations elucidated that this result was caused by phonon scattering by lattice defects as additives rather than the phonon–phonon interaction, and that the lattice strain dominantly contributed to it.
- Published
- 2014
31. Tissue-specific and ubiquitous gene knockouts by TALEN electroporation provide new approaches to investigating gene function in Ciona
- Author
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Takashi Yamamoto, Nicholas Treen, Yasunori Sasakura, Keita Yoshida, Tetsushi Sakuma, Haruka Sasaki, and Narudo Kawai
- Subjects
animal structures ,Molecular Sequence Data ,Mutant ,Computational biology ,Gene Knockout Techniques ,Animals ,Tissue Distribution ,Ciona intestinalis ,Molecular Biology ,Gene knockout ,Genetics ,Transcription activator-like effector nuclease ,Deoxyribonucleases ,Base Sequence ,biology ,Electroporation ,fungi ,Gene Expression Regulation, Developmental ,Gene targeting ,DNA ,biology.organism_classification ,Reverse genetics ,Ciona ,Mutagenesis ,Genetic Engineering ,Developmental Biology - Abstract
Custom designed nucleases can simplify gene targeting experiments and have the potential to allow these techniques to be performed in a wide range of organisms. Transcriptional activator-like effector nucleases (TALENs) are starting to fulfill this potential with the advantages of low cost and fast construction times. Here, we report that TALENs are highly effective at inducing mutations in specific genomic loci in the ascidian chordate Ciona intestinalis. In Ciona there are well-established methods to introduce exogenous DNA by electroporation, and we show that this method can be used to introduce constructs that can express TALENs ubiquitously or in specific tissues. Our current protocols enable the rapid analysis of hundreds of TALEN-induced mutants. TALEN electroporations result in a high rate of mutations. These mutations can result in gene knockouts that recapitulate previously described functions of Fgf3 and Hox12. We show that TALENs can work efficiently to cause tissue-specific knockouts and demonstrate this by knocking out Hox12 in the epidermis and Fgf3 in neural tissues. We also use tissue-specific knockouts to reveal a new function of Fgf3 during ascidian larval metamorphosis.
- Published
- 2014
32. Germ cell regeneration-mediated, enhanced mutagenesis in the ascidian Ciona intestinalis reveals flexible germ cell formation from different somatic cells
- Author
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Maki Shirae-Kurabayashi, Akiko Hozumi, Nicholas Treen, Takashi Yamamoto, Keita Yoshida, Yasunori Sasakura, and Tetsushi Sakuma
- Subjects
0301 basic medicine ,Male ,Somatic cell ,Germline ,Animals, Genetically Modified ,Gene Knockout Techniques ,0302 clinical medicine ,Mutation Rate ,Intestinal Mucosa ,Genetics ,education.field_of_study ,Transcription activator-like effector nuclease ,Genome ,Metamorphosis, Biological ,Spermatozoa ,Cell biology ,Ciona intestinalis ,Intestines ,medicine.anatomical_structure ,Electroporation ,Organ Specificity ,embryonic structures ,Germ line development ,Germ cell ,Tail ,endocrine system ,Population ,Biology ,03 medical and health sciences ,Transcription Activator-Like Effector Nucleases ,medicine ,Animals ,Regeneration ,education ,Molecular Biology ,Ovum ,Homeodomain Proteins ,Base Sequence ,urogenital system ,fungi ,Cell Biology ,biology.organism_classification ,Ciona ,030104 developmental biology ,Germ Cells ,Mutagenesis ,Mutation ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
The ascidian Ciona intestinalis has a high regeneration capacity that enables the regeneration of artificially removed primordial germ cells (PGCs) from somatic cells. We utilized PGC regeneration to establish efficient methods of germ line mutagenesis with transcription activator-like effector nucleases (TALENs). When PGCs were artificially removed from animals in which a TALEN pair was expressed, somatic cells harboring mutations in the target gene were converted into germ cells, this germ cell population exhibited higher mutation rates than animals not subjected to PGC removal. PGC regeneration enables us to use TALEN expression vectors of specific somatic tissues for germ cell mutagenesis. Unexpectedly, cis elements for epidermis, neural tissue and muscle could be used for germ cell mutagenesis, indicating there are multiple sources of regenerated PGCs, suggesting a flexibility of differentiated Ciona somatic cells to regain totipotency. Sperm and eggs of a single hermaphroditic, PGC regenerated animal typically have different mutations, suggesting they arise from different cells. PGCs can be generated from somatic cells even though the maternal PGCs are not removed, suggesting that the PGC regeneration is not solely an artificial event but could have an endogenous function in Ciona. This study provides a technical innovation in the genome-editing methods, including easy establishment of mutant lines. Moreover, this study suggests cellular mechanisms and the potential evolutionary significance of PGC regeneration in Ciona.
- Published
- 2016
33. A Case With Deteriorating Palmoplantar Pustulosis and Hyperthyroidism After Simultaneous Bimaxillary Orthognathic Surgery
- Author
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Aya Oda, Keita Yoshida, Taiga Yoshinaka, Akari Mukai, Tamayo Uno, and Masahiro Irifune
- Subjects
Adult ,congenital, hereditary, and neonatal diseases and abnormalities ,endocrine system ,medicine.medical_specialty ,Palmoplantar pustulosis ,endocrine system diseases ,medicine.medical_treatment ,Orthognathic surgery ,Hyperthyroidism ,Orthognathic Surgical Procedures ,Psoriasis ,otorhinolaryngologic diseases ,Hypersensitivity ,Humans ,Medicine ,business.industry ,medicine.disease ,Dermatology ,stomatognathic diseases ,Anesthesiology and Pain Medicine ,Anesthesia ,Female ,Brief Communications ,business - Abstract
A case of palmoplantar pustulosis and hyperthyroidism following orthognathic surgery is presented. Both diseases may have been related to allergic phenomena.
- Published
- 2017
34. Oxygen potential of hypo-stoichiometric La-doped UO2
- Author
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Kazuya Idemitsu, Keita Yoshida, Shuhei Miwa, Tatsumi Arima, Masahiko Osaka, and Yaohiro Inagaki
- Subjects
Lanthanide ,Nuclear and High Energy Physics ,Ionic radius ,Chemistry ,Uranium dioxide ,Analytical chemistry ,chemistry.chemical_element ,Uranium ,Atmospheric temperature range ,Oxygen ,chemistry.chemical_compound ,Nuclear Energy and Engineering ,Uranium oxide ,General Materials Science ,Stoichiometry ,Nuclear chemistry - Abstract
The oxygen potential of hypo-stoichiometric (U0.8La0.2)O2−x was measured as a function of oxygen-to-metal ratio (2 − x) in the temperature range from 1173 K to 1473 K. Compared with uranium oxides doped with other lanthanides, e.g. Lu, Er, Gd and Nd, the relationship between Δ G ¯ O 2 and the oxygen-to-metal ratio obtained for this sample showed almost the same trend, and for the relatively large oxygen non-stoichiometry x, its significance was comparable with that of (U0.8Gd0.2)O2−x. However, the detailed variation of Δ H ¯ O 2 and Δ S ¯ O 2 was different among these uranium oxides, especially in the near O/M = 2.0 region. Furthermore, the oxygen defect structure was evaluated from the relationship between log P O 2 and log x, and Δ G ¯ O 2 value of lanthanide-doped uranium oxide was discussed in terms of lanthanide ionic radius.
- Published
- 2011
35. Molecular Dynamics Study on Grain Boundary Diffusion of Actinides and Oxygen in Oxide Fuels
- Author
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Tatsumi Arima, Kazuya Idemitsu, Masahiro Nishina, Isamu Sato, Keita Yoshida, and Yaohiro Inagaki
- Subjects
Molecular dynamics ,chemistry.chemical_compound ,Chemistry ,Nuclear engineering ,Monte Carlo method ,Oxide ,Grain boundary diffusion coefficient ,Grain boundary ,General Medicine ,Actinide ,Diffusion (business) ,Supercomputer ,Nuclear chemistry - Published
- 2011
36. Repression of Rx gene on the left side of the sensory vesicle by Nodal signaling is crucial for right-sided formation of the ocellus photoreceptor in the development of Ciona intestinalis
- Author
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Hidetoshi Saiga and Keita Yoshida
- Subjects
Male ,Embryo, Nonmammalian ,Nodal Protein ,Green Fluorescent Proteins ,Cell ,Nodal signaling ,Nodal ,Sensory system ,Nervous System ,medicine ,Animals ,Ciona intestinalis ,Gene ,Psychological repression ,Molecular Biology ,In Situ Hybridization ,Body Patterning ,Homeodomain Proteins ,Microscopy, Confocal ,biology ,Reverse Transcriptase Polymerase Chain Reaction ,Gene Expression Regulation, Developmental ,Left–right asymmetry ,Embryo ,Anatomy ,Rx ,Cell Biology ,biology.organism_classification ,Cell biology ,medicine.anatomical_structure ,Ocellus ,Female ,Photoreceptor Cells, Invertebrate ,NODAL ,Signal Transduction ,Developmental Biology - Abstract
Nodal signaling plays an essential role in the establishment of left–right asymmetry in various animals. However, it is largely unknown how Nodal signaling is involved in the establishment of the left–right asymmetric morphology. In this study, the role of Nodal signaling in the left–right asymmetric ocellus formation in the ascidian, Ciona intestinalis was dealt with. During the development of C. intestinalis , the ocellus pigment cell forms on the midline and moves to the right side of the midline. Then, the photoreceptor cells form on the right side of the sensory vesicle (SV). Ci-Nodal is expressed on the left side of the SV in the developing tail bud embryo. When Nodal signaling is inhibited, the ocellus pigment cell form but remain on the midline, and expression of marker genes of the ocellus photoreceptor cells is ectopically detected on the left side as well as on the right side of the SV in the larva. Furthermore, Ci-Rx , which is essential for the ocellus differentiation, turns out to be negatively regulated by the Nodal signaling on the left side of the SV, even though it is required for the right-sided photoreceptor formation. These results indicate that Nodal signaling controls the left–right asymmetric ocellus formation in the development of C. intestinalis .
- Published
- 2011
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37. Platinum(II) and Platinum(IV) Porphyrin Pincer Complexes: Synthesis, Structures, and Reactivity
- Author
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Shigeru Yamaguchi, Hiroshi Shinokubo, Atsuhiro Osuka, and Keita Yoshida
- Subjects
Valence (chemistry) ,Organic Chemistry ,chemistry.chemical_element ,Bond formation ,Photochemistry ,Porphyrin ,Reductive elimination ,Pincer movement ,Inorganic Chemistry ,chemistry.chemical_compound ,chemistry ,Polymer chemistry ,Physical and Theoretical Chemistry ,Platinum ,Electronic properties ,Coordination geometry - Abstract
Pt(II) and Pt(IV) porphyrin pincer complexes were synthesized and characterized. Their structures and electronic properties were influenced by the valence state of platinum at the outer coordination site. The considerable distortion in the porphyrin core of the Pt(II) complex was relieved in the Pt(IV) complex, due to the change of the coordination geometry of platinum. Oxidation of the phenylplatinum(II) pincer complex with iodine induced a facile carbon−carbon bond formation reaction at the meso position via reductive elimination.
- Published
- 2010
38. Left–right asymmetric expression of Pitx is regulated by the asymmetric Nodal signaling through an intronic enhancer in Ciona intestinalis
- Author
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Hidetoshi Saiga and Keita Yoshida
- Subjects
endocrine system ,Embryo, Nonmammalian ,Morpholino ,Nodal Protein ,Nodal signaling ,Biology ,Nodal Signaling Ligands ,Transcription (biology) ,Genetics ,Animals ,Paired Box Transcription Factors ,Tissue Distribution ,Ciona intestinalis ,Genes, Developmental ,Enhancer ,Gene ,Conserved Sequence ,Body Patterning ,Regulation of gene expression ,Gene Expression Regulation, Developmental ,biology.organism_classification ,Molecular biology ,Introns ,Enhancer Elements, Genetic ,Epidermis ,NODAL ,Signal Transduction ,Developmental Biology - Abstract
Left-right asymmetric expression of Pitx as well as of Nodal has been observed in some ascidian species, but a mechanism that regulates the asymmetric expression of Pitx remains largely unclear. We addressed the transcription regulatory mechanism of the left-right asymmetric expression of Pitx gene in the ascidian, Ciona intestinalis. We first identified an intronic enhancer that drives Ci-Pitx left-sided expression in the epidermis and found that a single FoxH1 binding site present in this enhancer is essential for its activity. Also, we have shown that the enhancer requires Nodal signaling to drive the Ci-Pitx expression in the left epidermis. In C. intestinalis, left-sided expression of the Nodal gene has not been reported so far. We have confirmed that Ci-Nodal is expressed in the left epidermis of embryos at the tailbud stage only when they are allowed to develop within the chorion. Then, to test the importance of the FoxH1 binding site, we carried out knock down experiments using morpholino antisense oligonucleotides against Ci-FoxHa, the only FoxH gene that is expressed in the same stage as the left-sided expression of Ci-Pitx is observed. Knocking down of the function of Ci-FoxHa led to the down regulation of the expression of Ci-Pitx in the left epidermis. The present results suggest that the regulatory mechanism controlling the left-right asymmetric expression of the Pitx genes is well conserved between C. intestinalis and vertebrates.
- Published
- 2008
39. Nonreproductive role of gonadotropin-releasing hormone in the control of ascidian metamorphosis
- Author
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Chisato, Kamiya, Naoyuki, Ohta, Yosuke, Ogura, Keita, Yoshida, Takeo, Horie, Takehiro G, Kusakabe, Honoo, Satake, and Yasunori, Sasakura
- Subjects
Gonadotropin-Releasing Hormone ,Larva ,Cyclic AMP ,Metamorphosis, Biological ,Animals ,Cell Cycle Checkpoints ,Ciona intestinalis - Abstract
Gonadotropin-releasing hormones (GnRHs) are neuropeptides that play central roles in the reproduction of vertebrates. In the ascidian Ciona intestinalis, GnRHs and their receptors are expressed in the nervous systems at the larval stage, when animals are not yet capable of reproduction, suggesting that the hormones have non-reproductive roles.We showed that GnRHs in Ciona are involved in the animal's metamorphosis by regulating tail absorption and adult organ growth. Absorption of the larval tail and growth of the adult organs are two major events in the metamorphosis of ascidians. When larvae were treated with GnRHs, they completed tail absorption more frequently than control larvae. cAMP was suggested to be a second messenger for the induction of tail absorption by GnRHs. tGnRH-3 and tGnRH-5 (the "t" indicates "tunicate") inhibited the growth of adult organs by arresting cell cycle progression in parallel with the promotion of tail absorption.This study provides new insights into the molecular mechanisms of ascidian metamorphosis conducted by non-reproductive GnRHs.
- Published
- 2014
40. Chondroitin 6-O-sulfotransferases are required for morphogenesis of the notochord in the ascidian embryo
- Author
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Jun, Nakamura, Keita, Yoshida, Yasunori, Sasakura, and Shigeki, Fujiwara
- Subjects
Embryo, Nonmammalian ,Gene Knockdown Techniques ,Organogenesis ,Notochord ,Animals ,Gene Expression Regulation, Developmental ,Sulfotransferases ,Gene Expression Regulation, Enzymologic ,Ciona intestinalis - Abstract
Chondroitin sulfate (CS) is a sulfated polysaccharide chain that binds to various core proteins to form proteoglycans. The amount and position of sulfate groups in CS are variable among different tissues, and are determined by specific sulfotransferases. Although the ascidians are the closest relatives of vertebrates, the functions of their sulfotransferases have not been studied.The genome of the ascidian Ciona intestinalis contains eight genes encoding proteins similar to chondroitin 6-O-sulfotransferases (C6STs), which appear to have independently diverged in the ascidian lineage during evolution. Among them, Ci-C6ST-like1 and Ci-C6ST-like7 were predominantly expressed in the developing notochord. In addition, they were weakly expressed in the neural tube. The disruption of either one of them affected the convergent extension movement of notochordal cells. Presumptive notochord cells coming from both sides of the embryo did not intercalate. The results suggest that both of them are necessary. In some cases, the anterior neural tube failed to close. Forced expression of Ci-C6ST-like1 or Ci-C6ST-like7 in the notochord restored the normal intercalation of notochordal cells, indicating that the effects of morpholino oligos are specific.Ci-C6ST-like1 and Ci-C6ST-like7 are required for the morphogenesis of the notochord in the ascidian embryo.
- Published
- 2014
41. Application of detergents or high hydrostatic pressure as decellularization processes in uterine tissues and their subsequent effects on in vivo uterine regeneration in murine models
- Author
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Takashi Ushida, Akio Kishida, Keita Yoshida, Katsuko S. Furukawa, Yasushi Hirota, Shigeru Saito, Yutaka Osuga, Osamu Yoshino, Masanori Aizawa, and Erna G Santoso
- Subjects
Pathology ,medicine.medical_specialty ,Hydrostatic pressure ,Materials Science ,Detergents ,Uterus ,Biomedical Engineering ,lcsh:Medicine ,Bioengineering ,Extracellular matrix ,Andrology ,Biomaterials ,Rats, Sprague-Dawley ,Tissue engineering ,In vivo ,Pregnancy ,medicine ,Hydrostatic Pressure ,Animals ,Regeneration ,lcsh:Science ,Cells, Cultured ,Multidisciplinary ,Decellularization ,biology ,Tissue Engineering ,Tissue Scaffolds ,Chemistry ,lcsh:R ,Biology and Life Sciences ,Sodium Dodecyl Sulfate ,Extracellular Matrix ,Rats ,Transplantation ,medicine.anatomical_structure ,Fertility ,Physical Sciences ,biology.protein ,Engineering and Technology ,Female ,lcsh:Q ,Elastin ,Research Article ,Biotechnology - Abstract
Infertility caused by ovarian or tubal problems can be treated using In Vitro Fertilization and Embryo Transfer (IVF-ET); however, this is not possible for women with uterine loss and malformations that require uterine reconstruction for the treatment of their infertility. In this study, we are the first to report the usefulness of decellularized matrices as a scaffold for uterine reconstruction. Uterine tissues were extracted from Sprague Dawley (SD) rats and decellularized using either sodium dodecyl sulfate (SDS) or high hydrostatic pressure (HHP) at optimized conditions. Histological staining and quantitative analysis showed that both SDS and HHP methods effectively removed cells from the tissues with, specifically, a significant reduction of DNA contents for HHP constructs. HHP constructs highly retained the collagen content, the main component of extracellular matrices in uterine tissue, compared to SDS constructs and had similar content levels of collagen to the native tissue. The mechanical strength of the HHP constructs was similar to that of the native tissue, while that of the SDS constructs was significantly elevated. Transmission electron microscopy (TEM) revealed no apparent denaturation of collagen fibers in the HHP constructs compared to the SDS constructs. Transplantation of the decellularized tissues into rat uteri revealed the successful regeneration of the uterine tissues with a 3-layer structure 30 days after the transplantation. Moreover, a lot of epithelial gland tissue and Ki67 positive cells were detected. Immunohistochemical analyses showed that the regenerated tissues have a normal response to ovarian hormone for pregnancy. The subsequent pregnancy test after 30 days transplantation revealed successful pregnancy for both the SDS and HHP groups. These findings indicate that the decellularized matrix from the uterine tissue can be a potential scaffold for uterine regeneration.
- Published
- 2014
42. Transcriptional regulation of a horizontally transferred gene from bacterium to chordate
- Author
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Takashi Yamamoto, Rui Yokomori, Nicholas Treen, Tetsushi Sakuma, Keita Yoshida, Kenta Nakai, Sung-Joon Park, Yosuke Ogura, Yasunori Sasakura, Hidetoshi Saiga, and Shigeki Fujiwara
- Subjects
0301 basic medicine ,Gene Transfer, Horizontal ,Chordate ,Genome ,General Biochemistry, Genetics and Molecular Biology ,Actinobacteria ,03 medical and health sciences ,0302 clinical medicine ,Transcriptional regulation ,Animals ,Urochordata ,Gene ,Transcription factor ,Phylogeny ,Research Articles ,General Environmental Science ,Genetics ,Binding Sites ,General Immunology and Microbiology ,biology ,General Medicine ,biology.organism_classification ,Biological Evolution ,Tunicate ,030104 developmental biology ,Gene Expression Regulation ,Transcription Factor AP-2 ,Glucosyltransferases ,Horizontal gene transfer ,General Agricultural and Biological Sciences ,030217 neurology & neurosurgery - Abstract
The horizontal transfer of genes between distantly related organisms is undoubtedly a major factor in the evolution of novel traits. Because genes are functionless without expression, horizontally transferred genes must acquire appropriate transcriptional regulations in their recipient organisms, although the evolutionary mechanism is not known well. The defining characteristic of tunicates is the presence of a cellulose containing tunic covering the adult and larval body surface. Cellulose synthase was acquired by horizontal gene transfer from Actinobacteria. We found that acquisition of the binding site of AP-2 transcription factor was essential for tunicate cellulose synthase to gain epidermal-specific expression. Actinobacteria have very GC-rich genomes, regions of which are capable of inducing specific expression in the tunicate epidermis as the AP-2 binds to a GC-rich region. Therefore, the actinobacterial cellulose synthase could have been potentiated to evolve its new function in the ancestor of tunicates with a higher probability than the evolution depending solely on a spontaneous event.
- Published
- 2016
43. Germ cell mutations of the ascidian Ciona intestinalis with TALE nucleases
- Author
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Keita, Yoshida, Nicholas, Treen, Akiko, Hozumi, Tetsushi, Sakuma, Takashi, Yamamoto, and Yasunori, Sasakura
- Subjects
Gene Knockout Techniques ,Deoxyribonucleases ,Electroporation ,Genome ,Germ Cells ,Mutagenesis ,Animals ,Ciona intestinalis - Abstract
Targeted mutagenesis of genes-of-interest, or gene-knockout, is a powerful method to address the functions of genes. Engineered nucleases have enabled this approach in various organisms because of their ease of use. The ascidian Ciona intestinalis is an excellent organism to analyze gene functions by means of genetic technologies. In our previous study, we reported mutagenesis of Ciona somatic cells with TALE nucleases (TALENs) by electroporating expression constructs. In this study, we report germ cell mutagenesis of Ciona by microinjecting mRNAs encoding TALENs. TALEN mRNAs introduced mutations to target genes in both somatic and germ cells. TALEN-mediated mutations in the germ cell genome were inherited by the next generation. We conclude that knockout lines of Ciona that have disrupted target genes can be established through TALEN-mediated germ cell mutagenesis.
- Published
- 2013
44. Selective blockade of N-methyl-D-aspartate channels in combination with dopamine receptor antagonism induces loss of the righting reflex in mice, but not immobility
- Author
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Katsuya Morita, Nobuhito Kikuchi, Norio Sakai, Norimitsu Morioka, Yoshihiro Nakata, Yoshitaka Shimizu, Masahiro Irifune, Keita Yoshida, and Takashi Kanematsu
- Subjects
Agonist ,Male ,medicine.medical_specialty ,medicine.drug_class ,Receptors, N-Methyl-D-Aspartate ,chemistry.chemical_compound ,Immobilization ,Mice ,Reflex, Righting ,Internal medicine ,Haloperidol ,Medicine ,Animals ,Dexmedetomidine ,Pharmacology ,SCH-23390 ,business.industry ,Effective dose (pharmacology) ,Drug Combinations ,Endocrinology ,chemistry ,Reflex ,NMDA receptor ,Dopamine Antagonists ,Righting reflex ,Dizocilpine Maleate ,business ,Excitatory Amino Acid Antagonists ,medicine.drug - Abstract
The selective N-methyl-d-aspartate (NMDA) channel blocker MK-801 is known to induce no loss of the righting reflex (LORR) and to stimulate catecholaminergic (CAergic) neurons in rodents, playing a crucial role in arousal. We examined whether MK-801 in combination with CA receptor ligands, which inhibit CAergic neuronal activities, could induce anesthesia including LORR. All drugs were administered systemically to mice. To assess anesthesia, three different behaviors were used: loss of nociceptive response (analgesia in the free-moving state without LORR), LORR, and loss of movement in response to noxious stimulation (immobility under LORR). A very large dose of MK-801 (50 mg/kg) induced neither analgesia nor LORR. In contrast, MK-801 in combination with a small dose of the dopamine (DA) receptor antagonist haloperidol (0.2 mg/kg) dose-dependently produced LORR with a 50 % effective dose (ED50) of 1.6 (0.9–3.0; 95 % confidence limit) mg/kg, but not immobility. The α2-adrenoceptor agonist dexmedetomidine induced not only analgesia, but also immobility in animals treated with MK-801 (5 mg/kg) plus haloperidol (0.2 mg/kg), which then lost their righting reflex. The ED50 value of 0.26 (0.10–0.66) mg/kg (various doses of dexmedetomidine plus a fixed dose of MK-801 and haloperidol) for immobility was approximately three-fold larger than that of 0.09 (0.03–0.23) mg/kg (dexmedetomidine plus vehicle saline) for analgesia. This may occur, as LORR induced by MK-801 plus haloperidol inhibits the pain suppression system. The other ligands had little or no effect. The DAergic stimulant actions of MK-801 may mask its LORR effects by NMDA channel blockade.
- Published
- 2013
45. Uterine tissue engineering using a decellularized scaffold by hydrostatic pressure
- Author
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Katsuko, Furukawa, primary, Erna, Santoso, additional, Keita, Yoshida, additional, Yasushi, Hirota, additional, Osamu, Yoshino, additional, Akio, Kishida, additional, Takehiro, Hiraoka, additional, Yutaka, Osuga, additional, Shigeru, Saito, additional, and Takashi, Ushida, additional
- Published
- 2016
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- View/download PDF
46. 2A46 Study on reconstruction method of rat uterus using decellularized matrix scaffold
- Author
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Yasushi Hirota, Takashi Ushida, Osamu Yoshino, Katsuko Furukawa, Aiko Nakayasu, Keita Yoshida, Morikazu Seki, Yutaka Osuga, Akio Kishida, and Shigeru Saito
- Subjects
Scaffold ,Materials science ,Decellularized matrix ,Rat uterus ,Reconstruction method ,Biomedical engineering - Published
- 2015
47. 1H25 Uterus Tissue Engineering by the matrices decellularized with hyper hydrostatic pressure
- Author
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Shigeru Saitoh, Keita Yoshida, Yasushi Hirota, Jeonghyun Kim, Yutaka Osuga, Katsuko Furukawa, Osamu Yoshino, Akio Kishida, Morikazu Seki, Hiroshi Komatsu, and Takashi Ushida
- Subjects
Decellularization ,Materials science ,medicine.anatomical_structure ,Tissue engineering ,Hydrostatic pressure ,Uterus ,medicine ,Anatomy - Published
- 2016
48. Transcription regulatory mechanism of Pitx in the papilla-forming region in the ascidian, Halocynthia roretzi, implies conserved involvement of Otx as the upstream gene in the adhesive organ development of chordates
- Author
-
Hidetoshi Saiga, Keita Yoshida, Motoko Ueno, and Tomoko Niwano
- Subjects
endocrine system ,Morpholino ,Microinjections ,Transcription, Genetic ,Xenopus ,Notochord ,Embryonic Development ,Biology ,Conserved sequence ,Morpholinos ,Species Specificity ,Transcription (biology) ,Genes, Reporter ,medicine ,Animals ,Paired Box Transcription Factors ,RNA, Messenger ,Regulatory Elements, Transcriptional ,Urochordata ,Enhancer ,Gene ,Conserved Sequence ,Genetics ,Binding Sites ,Otx Transcription Factors ,Base Sequence ,Animal Structures ,Gene Expression Regulation, Developmental ,Cell Biology ,biology.organism_classification ,medicine.anatomical_structure ,Neurula ,Gene Knockdown Techniques ,Developmental Biology - Abstract
Pitx genes play important roles in a variety of developmental processes in vertebrates. In an ascidian species, Halocynthia roretzi, Hr-Pitx, the only Pitx gene of this species, has been reported to be expressed in the left epidermis at the tailbud stage. In the present study, first, we have shown that Hr-Pitx is also expressed in the papilla-forming region at the neurula to tailbud stages, and then we addressed transcription regulatory mechanisms for the expression of Hr-Pitx in the papilla-forming region. We have identified the genomic region ranging from 850 to 1211 bp upstream from the translation start site of the Hr-Pitx gene as an enhancer region that drives the transcription of Hr-Pitx in the papilla-forming region. Within the enhancer region, putative transcriptional factor binding sites for Otx as well as Fox were shown to be required for its activity. Finally, we carried out knocking down experiments of Hr-Otx function using an antisense morpholino oligonucleotide, in which the knocking down of Hr-Otx function resulted in reduction of the enhancer activity and loss of the expression of Hr-Pitx in the papilla-forming region. In Xenopus laevis, it has been reported that Pitx genes are expressed downstream of Otx function during development of the cement gland, an adhesive organ of its larva. Taken together, it is suggested that the expression regulatory mechanism of Pitx, involving Otx as the upstream gene, in the developing adhesive organ is conserved between ascidians and vertebrates.
- Published
- 2012
49. 2D16 Reconstruction of rat uterine tissue using a decellularized tissue scaffold
- Author
-
Jun Negishi, Osamu Yoshino, Takashi Ushida, Yutaka Osuga, Akio Kishida, Katsuko Furukawa, Shigeru Saito, Keita Yoshida, Yasushi Hirota, and Erna G Santoso
- Subjects
Scaffold ,Decellularization ,Chemistry ,Uterine tissue ,Biomedical engineering - Published
- 2014
50. Nodal signaling is involved in left-right asymmetric ocellus formation in Ciona intestinalis
- Author
-
Hidetoshi Saiga and Keita Yoshida
- Subjects
animal structures ,biology ,embryonic structures ,Nodal signaling ,Ciona intestinalis ,Cell Biology ,biology.organism_classification ,Molecular Biology ,Cell biology ,Developmental Biology - Published
- 2010
- Full Text
- View/download PDF
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