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164 results on '"Kusano, Kanichi"'

6. First evidence of the incorporation of daprodustat and other hypoxia-inducible factor stabilizers into equine hair by passive transfer based on segmental quantitative analysis

Author

Ishii, Hideaki, primary, Shibuya, Mariko, additional, Kusano, Kanichi, additional, Sone, Yu, additional, Kamiya, Takahiro, additional, Wakuno, Ai, additional, Ito, Hideki, additional, Miyata, Kenji, additional, Yamada, Masayuki, additional, and Leung, Gary Ngai-Wa, additional

Published
2023
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9. Segmental analysis and long-term monitoring of vadadustat in equine hair for the purpose of doping control.

Author

Ishii, Hideaki, Shibuya, Mariko, Kusano, Kanichi, Sone, Yu, Kamiya, Takahiro, Wakuno, Ai, Ito, Hideki, Miyata, Kenji, Yamada, Masayuki, and Leung, Gary Ngai-Wa

Subjects
HAIR analysis, HAIR growth, HAIR, SOLID phase extraction, LIQUID-liquid extraction, HYPOXIA-inducible factors, HORSE paces, gaits, etc., LIQUID chromatography-mass spectrometry
Abstract

Vadadustat is a newly launched hypoxia-inducible factor stabilizer with anti-anemia and erythropoietic effects; however, its use in horses is expressly forbidden in both racing and equestrian competitions. Following our previous report on the pharmacokinetic study of vadadustat in horse plasma and urine, a long-term longitudinal analysis of vadadustat in horse hair after nasoesophageal administration (3 g/day for 3 days) to three thoroughbred mares is described in this study. Our main objective is to further extend the detection period of vadadustat for the purpose of doping control. Three bunches of mane hair from each horse were collected at 0 (pre), 1, 2, 3 and 6 month(s) post-administration. These hair samples were each cut into 2-cm segments and pulverized after decontamination of hair samples. The analyte in the powdered hair samples was extracted with liquid–liquid extraction followed by further purification by solid-phase extraction with strong anion exchange columns. The amount of vadadustat incorporated into the hair was quantified with a newly developed and validated method using liquid chromatography–high-resolution mass spectrometry. Our results show that vadadustat was confirmed in all post-administration hair samples, but its metabolites were not present. Thus, the detection window for vadadustat could be successfully extended up to 6 months post-administration. Interestingly, the 2-cm segmental analysis revealed that the tip of the drug band in the hair shifted along with the hair shafts in correspondence with the average hair growth rate (∼2.5 cm/month) but gradually diffused more widely from 2 cm at 1 month post-administration to up to 14 cm at 6 months post-administration. However, the loss in the total amount of vadadustat in hair over time was observed to most likely be due to the degradation of vadadustat. These findings will be useful for the control of abuse and/or misuse of vadadustat and the interpretation of positive doping cases. [ABSTRACT FROM AUTHOR]

Published
2023
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12. Additional studies on nicotine exposure in horses: Accurate quantification and elimination profiles of potential biomarkers in plasma and urine

Author

Ishii, Hideaki, primary, Shibuya, Mariko, additional, Leung, Gary Ngai‐Wa, additional, Yamashita, Shozo, additional, Nagata, Shun‐ichi, additional, Kushiro, Asuka, additional, Sakai, Satoshi, additional, Toju, Kota, additional, Okada, Jun, additional, Kawasaki, Kazumi, additional, Kusano, Kanichi, additional, and Kijima‐Suda, Isao, additional

Published
2022
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13. Unique insertion/deletion polymorphisms within histidine-rich region of histidine-rich glycoprotein in Thoroughbred horses

Author

Muko, Ryo, primary, Sunouchi, Tomoya, additional, Urayama, Shuntaro, additional, Toishi, Yuko, additional, Kusano, Kanichi, additional, Sato, Hiroaki, additional, Muranaka, Masanori, additional, Shin, Taekyun, additional, Oikawa, Masa-aki, additional, Ojima, Yoshinobu, additional, Ali, Mohammad, additional, Nomura, Yoshihiro, additional, Matsuda, Hiroshi, additional, and Tanaka, Akane, additional

Published
2022
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15. Pharmacokinetic Study of Vadadustat and High-Resolution Mass Spectrometric Characterization of its Novel Metabolites in Equines for the Purpose of Doping Control

Author

Ishii, Hideaki, primary, Shibuya, Mariko, additional, Kusano, Kanichi, additional, Sone, Yu, additional, Kamiya, Takahiro, additional, Wakuno, Ai, additional, Ito, Hideki, additional, Miyata, Kenji, additional, Sato, Fumio, additional, Kuroda, Taisuke, additional, Yamada, Masayuki, additional, and Leung, Gary Ngai-Wa, additional

Published
2022
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16. Antimicrobial resistance profiles and phylogenetic groups of Escherichia coli isolated from healthy Thoroughbred racehorses in Japan

Author

SATO, Wataru, SUKMAWINATA, Eddy, UEMURA, Ryoko, KANDA, Takuya, KUSANO, Kanichi, KAMBAYASHI, Yoshinori, SATO, Takashi, ISHIKAWA, Yuhiro, TOYA, Ryohei, and SUEYOSHI, Masuo

Subjects
Full Paper, Escherichia coli, antimicrobial resistant, Thoroughbred
Abstract

In this study, we investigated the occurrence of antimicrobial resistance in commensal Escherichia coli isolated from healthy Thoroughbred (TB) racehorses in Japan. A total of 212 fecal samples were individually collected from TB racehorses from March 2017 to August 2018 at Japan Racing Association training centers. E. coli was isolated by using selective agar media, deoxycholate-hydrogen sulfide-lactose (DHL) and eosin methylene blue (EMB). A total of 417 E. coli isolates were examined against 10 antimicrobial agents by using the broth microdilution method. The 417 E. coli isolates were phylogenetically grouped using a multiplex polymerase chain reaction. The highest proportion of resistance was observed for streptomycin (30.9%, 129/417) followed by ampicillin (19.4%, 81/417), trimethoprim (15.8%, 66/417), tetracycline (8.4%, 35/417), chloramphenicol (2.6%, 11/417), kanamycin (1.2%, 5/417), nalidixic acid (0.5%, 2/417), cefazolin (0.2%, 1/417), colistin (0.2%, 1/417), and gentamycin (0%). Multidrug-resistant (MDR) E. coli was detected in 7.9% (33/417) of isolates. The proportions of resistance against ampicillin, streptomycin, kanamycin, and chloramphenicol and of multidrug-resistant phenotypes in E. coli belonging to phylogenetic group B2 were significantly higher than those of other groups. This study clarified the distribution of antimicrobial-resistant (AMR) E. coli in Japanese racehorses. A continuous monitoring program for antimicrobial resistance is required to control the spread of AMR bacteria in racehorses.

Published
2020

17. Long‐term monitoring of IOX4 in horse hair and its longitudinal distribution with segmental analysis using liquid chromatography/electrospray ionization Q Exactive high‐resolution mass spectrometry for the purpose of doping control

Author

Ishii, Hideaki, primary, Shibuya, Mariko, additional, So, Yat‐Ming, additional, Wong, Jenny K. Y., additional, Ho, Emmie N. M., additional, Kusano, Kanichi, additional, Sone, Yu, additional, Kamiya, Takahiro, additional, Wakuno, Ai, additional, Ito, Hideki, additional, Miyata, Kenji, additional, Yamada, Masayuki, additional, and Leung, Gary Ngai‐Wa, additional

Published
2022
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21. Medication control of flunixin in racing horses: Possible detection times using Monte Carlo simulations

Author

Kuroda, Taisuke, primary, Minamijima, Yohei, additional, Nomura, Motoi, additional, Yamashita, Shozo, additional, Yamada, Masayuki, additional, Nagata, Shunichi, additional, Mita, Hiroshi, additional, Tamura, Norihisa, additional, Fukuda, Kentaro, additional, Kuwano, Atsutoshi, additional, Kusano, Kanichi, additional, Toutain, Pierre‐Louis, additional, and Sato, Fumio, additional

Published
2021
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24. Comprehensive metabolic study of IOX4 in equine urine and plasma using liquid chromatography/electrospray ionization Q Exactive high‐resolution mass spectrometer for the purpose of doping control

Author

Ishii, Hideaki, primary, Shibuya, Mariko, additional, So, Yat‐Ming, additional, Wong, Jenny K. Y., additional, Ho, Emmie N. M., additional, Kusano, Kanichi, additional, Sone, Yu, additional, Kamiya, Takahiro, additional, Wakuno, Ai, additional, Ito, Hideki, additional, Miyata, Kenji, additional, Yamada, Masayuki, additional, and Leung, Gary Ngai‐Wa, additional

Published
2021
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26. Sequence determination of phosphorothioated oligonucleotides using MALDI‐TOF mass spectrometry for controlling gene doping in equestrian sports

Author

Tozaki, Teruaki, primary, Kwak, Ho‐Geun, additional, Nakamura, Kotono, additional, Takasu, Masaki, additional, Ishii, Hideaki, additional, Ohnuma, Aoi, additional, Kikuchi, Mio, additional, Ishige, Taichiro, additional, Kakoi, Hironaga, additional, Hirota, Kei‐ichi, additional, Kusano, Kanichi, additional, Hirata, Minoru, additional, Nirasawa, Takashi, additional, and Nagata, Shun‐ichi, additional

Published
2021
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28. Medication control of flunixin in racing horses: Possible detection times using Monte Carlo simulations.

Author

Kuroda, Taisuke, Minamijima, Yohei, Nomura, Motoi, Yamashita, Shozo, Yamada, Masayuki, Nagata, Shunichi, Mita, Hiroshi, Tamura, Norihisa, Fukuda, Kentaro, Kuwano, Atsutoshi, Kusano, Kanichi, Toutain, Pierre‐Louis, and Sato, Fumio

Abstract

Summary: Background: For medication control in several jurisdictions, withdrawal time is the period of refrain from racing after drug administration. It is set by adding a safety period to an experimental detection time. However, there are no reports of statistical analyses of detection time for the determination of withdrawal time in flunixin meglumine‐treated horses. Objective: To analyse the population pharmacokinetics of flunixin in horses through the generation of a dataset for detection time statistical analysis and predictions via Monte Carlo simulation. Study design: Experimental study. Methods: Drug plasma and urine concentrations following single intravenous administration of flunixin 1.1 mg/kg bodyweight (BW) in 10 horses and multiple administration of q 24 hours for 5 days in 10 horses were measured using liquid chromatography with tandem mass spectrometry (LC‐MS/MS). Data were modelled using a nonlinear mixed effect model followed by Monte Carlo simulation. Irrelevant plasma concentration (IPC) and irrelevant urine concentration (IUC) were calculated using the Toutain approach. Detection times were obtained considering the time after the last administration for selected quantiles of 5000 hypothetical horses under the international screening limit (ISL) proposed by the International Federation of Horseracing Authorities (plasma: 1 ng/mL, urine; 100 ng/mL). Results: For a regimen of 1.1 mg/kg BW q 24 hours, the IPC and IUC values were 2.0 and 73.0 ng/mL respectively. Detection times in plasma above the ISL for 90% of simulated horses were estimated as 74 hours after a single 1.1 mg/kg dose administration, 149 and 199 hours after multiple doses over 5 days at either 24‐ or 12‐hour intervals respectively. Corresponding detection times in urine were 46, 68 and 104 hours respectively. Main limitation: Only female horses were investigated. Conclusions: Statistical detection times for different flunixin meglumine regimens indicated a delay of detection time in plasma after multiple administrations under ISL. [ABSTRACT FROM AUTHOR]

Published
2022
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30. Gene Doping Control in Horseracing Industries

Author

TOZAKI, Teruaki, primary, OHNUMA, Aoi, additional, KIKUCHI, Mio, additional, ISHIGE, Taichiro, additional, HIROTA, Kei-ichi, additional, KAKOI, Hironaga, additional, KUSANO, Kanichi, additional, and NAGATA, Shun-ichi, additional

Published
2021
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38. Low‐copy transgene detection using nested digital polymerase chain reaction for gene‐doping control.

Author

Tozaki, Teruaki, Ohnuma, Aoi, Hamilton, Natasha A., Kikuchi, Mio, Ishige, Taichiro, Kakoi, Hironaga, Hirota, Kei‐ichi, Kusano, Kanichi, and Nagata, Shun‐ichi

Abstract

Gene doping is prohibited for fair competition in human and horse sports. One style of gene doping is the administration of an exogeneous gene, called a transgene, to postnatal humans and horses. Although many transgene detection methods based on quantitative polymerase chain reaction (PCR), including real‐time PCR and digital PCR, have been recently developed, it remains difficult to reliably detect low‐copy transgenes. In this study, we developed and validated a nested digital PCR method to specifically detect low‐copy transgenes. The nested digital PCR consists of (1) preamplification using conventional PCR and (2) droplet digital PCR detection using a hydrolysis probe. Using 5, 10, 20, 60 and 120 transgene copies as template, 496.0, 1089.7, 1820.7, 4313.3 and 7840.0 copies per microlitre, respectively, were detected using our nested digital PCR. Although high concentrations of phenol, proteinase K, ethanol, EDTA, heparin and genomic DNA all inhibited preamplification, their effects on the digital PCR detection were limited. Once preamplification was successful, even substitution of bases within the primers and probes had minimal effects on transgene detection. The nested digital PCR developed in this study successfully detected low‐copy transgenes and can be used to perform a qualitative test, indicating its usefulness in the prevention of false positives and false negatives in gene‐doping detection. [ABSTRACT FROM AUTHOR]

Published
2022
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39. Comprehensive metabolic study of IOX4 in equine urine and plasma using liquid chromatography/electrospray ionization Q Exactive high‐resolution mass spectrometer for the purpose of doping control.

Author

Ishii, Hideaki, Shibuya, Mariko, So, Yat‐Ming, Wong, Jenny K. Y., Ho, Emmie N. M., Kusano, Kanichi, Sone, Yu, Kamiya, Takahiro, Wakuno, Ai, Ito, Hideki, Miyata, Kenji, Yamada, Masayuki, and Leung, Gary Ngai‐Wa

Abstract

IOX4 is a hypoxia‐inducible factor prolyl hydroxylase (HIF‐PHD) inhibitor, which was developed for the treatment of anemia by exerting hematopoietic effects. The administration of HIF‐PHD inhibitors such as IOX4 to horses is strictly prohibited by the International Federation of Horseracing Authorities and the Fédération Équestre Internationale. To the best of our knowledge, this is the first comprehensive metabolic study of IOX4 in horse plasma and urine after a nasoesophageal administration of IOX4 (500 mg/day, 3 days). A total of four metabolites (three mono‐hydroxylated IOX4 and one IOX4 glucuronide) were detected from the in vitro study using homogenized horse liver. As for the in vivo study, post‐administration plasma and urine samples were comprehensively analyzed with liquid chromatography/electrospray ionization high‐resolution mass spectrometry to identify potential metabolites and determine their corresponding detection times. A total of 10 metabolites (including IOX4 glucuronide, IOX4 glucoside, O‐desbutyl IOX4, O‐desbutyl IOX4 glucuronide, four mono‐hydroxylated IOX4, N‐oxidized IOX4, and N‐oxidized IOX4 glucoside) were found in urine and three metabolites (glucuronide, glucoside, and O‐desbutyl) in plasma. Thus, the respective quantification methods for the detection of free and conjugated IOX4 metabolites in urine and plasma with a biphase enzymatic hydrolysis were developed and applied to post‐administration samples for the establishment of elimination profiles of IOX4. The detection times of total IOX4 in urine and plasma could be successfully prolonged to at least 312 h. [ABSTRACT FROM AUTHOR]

Published
2022
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40. Sequence determination of phosphorothioated oligonucleotides using MALDI‐TOF mass spectrometry for controlling gene doping in equestrian sports.

Author

Tozaki, Teruaki, Kwak, Ho‐Geun, Nakamura, Kotono, Takasu, Masaki, Ishii, Hideaki, Ohnuma, Aoi, Kikuchi, Mio, Ishige, Taichiro, Kakoi, Hironaga, Hirota, Kei‐ichi, Kusano, Kanichi, Hirata, Minoru, Nirasawa, Takashi, and Nagata, Shun‐ichi

Abstract

In human and equestrian sporting events, one method of gene doping is the illegal use of therapeutic oligonucleotides to alter gene expression. In this study, we aimed to identify therapeutic oligonucleotides via sequencing using matrix‐assisted laser desorption/ionisation‐time‐of‐flight mass spectrometry (MALDI‐TOF MS). As a model of therapeutic oligonucleotides, 22 bp‐long phosphorothioated oligonucleotides (PSOs) were used. By using a Clarity OTX kit for extracting short‐length oligonucleotides, a spectrum of singly charged PSO with a mean intensity of 6.08 × 104 (standard deviation: 4.34 × 103) was detected from 500 pmol PSO in 1 ml horse plasma using the linear negative mode of MALDI‐TOF MS. In addition, a 17 bp sequence was determined using in‐source decay (ISD) mode, indicating that 500 pmol of a PSO in 1 ml plasma is the detection limit for sequencing. Using the determined sequences (17 bp), a targeted gene for PSO was singly identified on the horse reference genome, EquCab2.0, via a GGGenome search. These procedures can be potentially used to identify therapeutic oligonucleotides, whose nucleotides are unknown, for gene doping control. [ABSTRACT FROM AUTHOR]

Published
2022
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49. A retrospective comparison of induction with thiopental/guaifenesin and propofol/ketamine in Thoroughbred racehorses anesthetized with sevoflurane and medetomidine during arthroscopic surgery

Author

TOKUSHIGE, Hirotaka, primary, ARAKI, Masanari, additional, KUSANO, Kanichi, additional, ARIMA, Daisuke, additional, ITO, Hideki, additional, YAMAZAKI, Yosuke, additional, URAYAMA, Shuntaro, additional, KAMBAYASHI, Yoshinori, additional, TATENO, Oki, additional, and OHTA, Minoru, additional

Published
2019
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