11 results on '"Hirama, Masahiro"'
Search Results
2. A study on mechanisms of toxic actions of ciguatoxins: existence of functional relationship between CTX3C and charged residues of voltage sensors in Nav1.4 sodium channel
- Author
-
Yamaoka, Kaoru, Inoue, Masayuki, and Hirama, Masahiro
- Published
- 2011
- Full Text
- View/download PDF
3. Rationally Designed Synthetic Haptens to Generate Anti-Ciguatoxin Monoclonal Antibodies, and Development of a Practical Sandwich ELISA to Detect Ciguatoxins.
- Author
-
Tsumuraya, Takeshi and Hirama, Masahiro
- Subjects
- *
MONOCLONAL antibodies , *FOOD poisoning , *HAPTENS , *SEAFOOD poisoning , *ENZYME-linked immunosorbent assay - Abstract
"Ciguatera" fish poisoning (CFP) is one of the well-known food poisoning caused by the ingestion of fish that have accumulated trace amounts of ciguatoxins (CTXs). CFP affects more than 50,000 individuals annually. The difficulty in preventing CFP comes from the lack of reliable methods for analysis of CTXs in contaminated fish, together with the normal appearance, taste, and smell of CTX-contaminated fish. Thus, a sensitive, accurate, routine, and portable analytical method to detect CTXs is urgently required. Monoclonal antibodies (mAbs) specific against either wing of major CTX congeners (CTX1B, 54-deoxyCTX1B, CTX3C, and 51-hydroxyCTX3C) were generated by immunizing mice with rationally designed synthetic haptens-KLH conjugates instead of the CTXs. Haptenic groups with a surface area greater than 400 Å2 are required to produce mAbs that can strongly bind to CTXs. Furthermore, a highly sensitive fluorescence-based sandwich enzyme-linked immunosorbent assay (ELISA) was developed. This protocol can detect and quantify four major CTX congeners (CTX1B, 54-deoxyCTX1B, CTX3C, and 51-hydroxyCTX3C) with a limit of detection (LOD) of less than 1 pg/mL. The LOD determined for this sandwich ELISA is sufficient to detect CTX1B-contaminated fish at the FDA guidance level of 0.01 ppb. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
4. Transcriptomic Analysis of Ciguatoxin-Induced Changes in Gene Expression in Primary Cultures of Mice Cortical Neurons.
- Author
-
Rubiolo, Juan Andrés, Vale, Carmen, Boente-Juncal, Andrea, Hirama, Masahiro, Yamashita, Shuji, Camiña, Mercedes, Vieytes, Mercedes R., and Botana, Luis M.
- Subjects
NEURONS ,TOXINS ,SODIUM channels ,GENES ,DNA microarrays - Abstract
Ciguatoxins are polyether marine toxins that act as sodium channel activators. These toxins cause ciguatera, one of the most widespread nonbacterial forms of food poisoning, which presents several symptoms in humans including long-term neurological alterations. Earlier work has shown that both acute and chronic exposure of primary cortical neurons to synthetic ciguatoxin CTX3C have profound impacts on neuronal function. Thus, the present work aimed to identify relevant neuronal genes and metabolic pathways that could be altered by ciguatoxin exposure. To study the effect of ciguatoxins in primary neurons in culture, we performed a transcriptomic analysis using whole mouse genome microarrays, for primary cortical neurons exposed during 6, 24, or 72 h in culture to CTX3C. Here, we have shown that the effects of the toxin on gene expression differ with the exposure time. The results presented here have identified several relevant genes and pathways related to the effect of ciguatoxins on neurons and may assist in future research or even treatment of ciguatera. Moreover, we demonstrated that the effects of the toxin on gene expression were exclusively consequential of its action as a voltage-gated sodium channel activator, since all the effects of CTX3C were avoided by preincubation of the neurons with the sodium channel blocker tetrodotoxin. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
5. Humanization and characterization of an anti-ciguatoxin CTX3C monoclonal antibody.
- Author
-
Yamaguchi, Asako, Aino, Hiroaki, Tsumuraya, Takeshi, Hirama, Masahiro, and Fujii, Ikuo
- Subjects
- *
MONOCLONAL antibodies , *FOODBORNE diseases , *IMMUNOGLOBULINS , *GERM cells , *ANTIGENS - Abstract
Ciguatera poisoning (CP), caused by ciguatoxins (CTXs), is one of the most common food-borne diseases, affecting more than 50,000 people each year. In most cases, CP are managed with symptomatic and supportive remedies, and no specific treatment has been devised. In this study, toward the development of therapeutic antibodies for CP, we examined to humanize mouse anti-CTX3C antibody 10C9 (m10C9), which exhibited neutralizing activity against ciguatoxin in vitro and in vivo. The complementarity determining regions were grafted onto a human germline sequence with high sequence identity to m10C9, and the backmutations were examined to maintain the binding affinity. The optimized humanized antibody, Opt.h10C9Fab, showed a strong binding affinity to CTX3C with a high affinity (K D = 19.0 nM), and only two backmutations of ArgL46 and CysH94 in the framework regions were involved in determining the antigen binding affinity. [Display omitted] • The mouse anti-ciguatoxin CTX3C antibody was humanized by CDR-grafting coupled with the backmutations. • The optimized humanized antibody maintained a high binding affinity to CTX3C. • Two backmutations in the framework regions were involved in antigen binding affinity. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
6. Production of monoclonal antibodies for sandwich immunoassay detection of Pacific ciguatoxins
- Author
-
Tsumuraya, Takeshi, Fujii, Ikuo, and Hirama, Masahiro
- Subjects
- *
MARINE toxins , *MONOCLONAL antibodies , *IMMUNOASSAY , *TOXICOLOGY of poisonous fishes , *SEAFOOD poisoning , *ENZYME-linked immunosorbent assay , *HAPTENS , *CROSS reactions (Immunology) - Abstract
Abstract: Ciguatoxins are the major causative toxins of ciguatera seafood poisoning. Limited availability of ciguatoxins has hampered the development of a reliable and specific immunoassay for detecting these toxins in contaminated fish. Monoclonal antibodies (mAbs) specific against both ends of Pacific ciguatoxins CTX3C and 51-hydroxyCTX3C were prepared by immunization of mice with the protein conjugates of rationally designed synthetic haptens in place of the natural toxin. Haptenic groups that possess a surface area larger than 400Å2 were required to produce mAbs that can bind strongly to CTX3C or 51-hydroxyCTX3C. A direct sandwich enzyme-linked immunosorbent assay (ELISA) using these mAbs was established to detect CTX3C and 51-hydroxyCTX3C at the ppb level with no cross-reactivity against the other marine toxins, including brevetoxin A, brevetoxin B, okadaic acid, or maitotoxin. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
7. Evaluation of relative potency of calibrated ciguatoxin congeners by near-infrared fluorescent receptor binding and neuroblastoma cell-based assays.
- Author
-
Yokozeki, Toshiaki, Hama, Yuka, Fujita, Kazuhiro, Igarashi, Tomoji, Hirama, Masahiro, and Tsumuraya, Takeshi
- Subjects
- *
BINDING site assay , *TOXAPHENE , *NEUROBLASTOMA , *SEAFOOD poisoning , *FISHERY resources , *FOODBORNE diseases , *SODIUM channels - Abstract
Ciguatera fish poisoning (CFP) is a foodborne illness affecting > 50,000 people worldwide annually. It is caused by eating marine invertebrates and fish that have accumulated ciguatoxins (CTXs). Recently, the risk of CFP to human health, the local economy, and fishery resources have increased; therefore, detection methods are urgently needed. Functional assays for detecting ciguatoxins in fish include receptor binding (RBA) and neuroblastoma cell-based assay (N2a assay), which can detect all CTX congeners. In this study, we made these assays easier to use. For RBA, an assay was developed using a novel near-infrared fluorescent ligand, PREX710-BTX, to save valuable CTXs. In the N2a assay, a 1-day assay was developed with the same detection performance as the conventional 2-day assay. Additionally, in these assays, we used calibrated CTX standards from the Pacific determined by quantitative NMR for the first time to compare the relative potency of congeners, which differed significantly among previous studies. In the RBA, there was almost no difference in the binding affinity among congeners, showing that the differences in side chains, stereochemistry, and backbone structure of CTXs did not affect the binding affinity. However, this result did not correlate with the toxic equivalency factors (TEFs) based on acute toxicity in mice. In contrast, the N2a assay showed a good correlation with TEFs based on acute toxicity in mice, except for CTX3C. These findings, obtained with calibrated toxin standards, provide important insights into evaluating the total toxicity of CTXs using functional assays. [Display omitted] • Receptor binding assay using near-infrared fluorescent-labeled brevetoxin was effective in saving valuable ciguatoxins. • Assay time of neuroblastoma cell-based assay was shortened by omitting the 24 h pre-culture without lowering detectability. • Differences in binding affinities of calibrated ciguatoxin congeners to voltage-gated sodium channels were slight. • The relative potency of neuroblastoma cell-based assay for ciguatoxin congeners correlated well with acute toxicity in mice. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
8. Development of a monoclonal antibody against the left wing of ciguatoxin CTX1B: Thiol strategy and detection using a sandwich ELISA
- Author
-
Tsumuraya, Takeshi, Takeuchi, Katsutoshi, Yamashita, Shuji, Fujii, Ikuo, and Hirama, Masahiro
- Subjects
- *
TOXICOLOGY of poisonous fishes , *MONOCLONAL antibodies , *THIOLS , *ENZYME-linked immunosorbent assay , *FOOD poisoning , *INGESTION , *REEF fishes , *DINOFLAGELLATES , *FOOD chains - Abstract
Abstract: Ciguatera fish poisoning (CFP) is a form of food poisoning caused by the ingestion of a variety of reef fish that have accumulated trace amounts of ciguatoxins produced by dinoflagellates of the genus Gambierdiscus through the food chain. CFP affects more than 50,000 people each year. The extremely low level of the causative neurotoxins, ciguatoxins, in fish has hampered the preparation of antibodies for detecting the toxins. In this paper, we describe a thiol strategy for synthesizing a keyhole limpet hemocyanin (KLH)-conjugate (20) of the ABCDE-ring fragment of the Pacific ciguatoxins, CTX1B (1) and 54-deoxyCTX1B (4). We succeeded in producing a monoclonal antibody (3G8) against the left wings of these ciguatoxins by immunizing mice with the hapten-KLH conjugate (20) as the synthetic antigen. The most promising mAb, 3G8, does not cross-react with other related marine toxins. Sandwich enzyme-linked immunosorbent assay (ELISA) utilizing 3G8 and the previously prepared monoclonal antibody (8H4) enabled us to detect 1 specifically at less than 0.28 ng/mL. [Copyright &y& Elsevier]
- Published
- 2012
- Full Text
- View/download PDF
9. Ciguatera incidence and fish toxicity in Okinawa, Japan
- Author
-
Oshiro, Naomasa, Yogi, Kentaro, Asato, Shuko, Sasaki, Toshiki, Tamanaha, Koji, Hirama, Masahiro, Yasumoto, Takeshi, and Inafuku, Yasuo
- Subjects
- *
TOXICOLOGY of poisonous fishes , *IDENTIFICATION of fishes , *BIOLOGICAL assay , *LUTJANUS , *EPINEPHELUS , *LIQUID chromatography , *MASS spectrometry - Abstract
Abstract: Okinawa being located in the subtropical region has the highest incidence of ciguatera in Japan. Officially, 33 outbreaks involving 103 patients have been reported between 1997 and 2006. The implicated species were Variola louti, Lutjanus bohar, Lutjanus monostigma, Epinephelus fuscoguttatus, unidentified Lutjanus sp., Plectropomus areolatus, Oplegnathus punctatus, Epinephelus polyphekadion, Caranx ignobilis and moray eel. Toxicities of the leftover meals, as determined by mouse bioassays, ranged from 0.025 to 0.8MU/g or above (equivalent to 0.175–5.6ngCTX1B/g). We collected 612 specimens of fish belonging to L. monostigma, L. bohar, Lutjanus argentimaculatus, Lutjanus russellii, V. louti, Variola albimarginata, and E. fuscoguttatus from the coasts around Okinawa and examined the toxicity of the flesh by the mouse bioassay. The rate of toxic fish was as follows: L. monostigma: 32.3%, L. bohar: 11.9%, V. louti: 14.3%, E. fuscoguttatus: 20.8%. Only one out of 36 samples of V. albimarginata and two of 74 samples of L. russellii were found toxic. None of the 35 samples of L. argentimaculatus was toxic. Nor the L. bohar samples weighing less than 4kg were toxic. In all toxic samples, CTX1B was detected by LC/MS analysis but CTX3C and 51-hydroxyCTX3C were not. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
10. Use of monoclonal antibodies as an effective strategy for treatment of ciguatera poisoning
- Author
-
Inoue, Masayuki, Lee, Nayoung, Tsumuraya, Takeshi, Fujii, Ikuo, and Hirama, Masahiro
- Subjects
- *
FOOD poisoning , *MONOCLONAL antibodies , *TOXICOLOGY of poisonous fishes , *FOODBORNE diseases , *TOXICOLOGICAL emergencies , *FORENSIC toxicology , *SODIUM channels , *THERAPEUTICS - Abstract
Abstract: Ciguatera is a global food poisoning caused by the consumption of fish that have accumulated sodium channel activator toxins, ciguatoxins. At present, most diagnosed cases of ciguatera are treated with symptomatic and supportive remedies, and no specific therapy has been devised. Here we report that ciguatoxin CTX3C can be effectively neutralized in vitro and in vivo by simultaneous use of two anti-ciguatoxin monoclonal antibodies, providing the first rational approach toward directly preventing and treating ciguatera. [Copyright &y& Elsevier]
- Published
- 2009
- Full Text
- View/download PDF
11. Production of monoclonal antibodies for sandwich immunoassay detection of ciguatoxin 51-hydroxyCTX3C
- Author
-
Tsumuraya, Takeshi, Fujii, Ikuo, Inoue, Masayuki, Tatami, Atsushi, Miyazaki, Keisuke, and Hirama, Masahiro
- Subjects
- *
SEAFOOD poisoning , *TOXICOLOGY of poisonous fishes , *NEUROTOXIC agents , *MONOCLONAL antibodies - Abstract
Abstract: Every year, more than 50,000 people in subtropical and tropical regions suffer from ciguatera seafood poisoning. The extremely low level of the causative neurotoxins (ciguatoxins) in fish has hampered the preparation of antibodies for detection of the toxins. In this study, we produced a monoclonal antibody (8H4) against the right end of ciguatoxin CTX1B (1) and 51-hydroxyCTX3C (3) by immunizing mice with the keyhole limpet hemocyanin-conjugate of the synthetic HIJKLM ring fragment (10). We used 8H4 and another previously reported monoclonal antibody (10C9) that recognizes the left end of 3 to develop a sandwich enzyme-linked immunosorbent assay (ELISA) to detect 3. The assay could detect 3 down to the ppb level and lacked cross-reactivity with other related marine toxins, including brevetoxin A, brevetoxin B, okadaic acid, and maitotoxin. [Copyright &y& Elsevier]
- Published
- 2006
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.