Dear Editor,Amplexus,a reproductive behavior in which the male clasps the female dorsally,is a characteristic of amphibians(Wells,2007).Most species exhibit either inguinal or axillary amplexus,although some reproduce...Dear Editor,Amplexus,a reproductive behavior in which the male clasps the female dorsally,is a characteristic of amphibians(Wells,2007).Most species exhibit either inguinal or axillary amplexus,although some reproduce without it(Wells,2007).While male-female amplexus is predominant,other forms such as male-male and multiple-male amplexus have also been documented(Soni et al.,2025).Release calls are typically produced by amplexed males to signal mating mismatches and mitigate the costs of inappropriate clasping(Kelehear and Shine,2019).Female-female amplexus,however,is rare and considered maladaptive,as it does not lead to fertilization(Chuang et al.,2019).展开更多
The combination of horseradish peroxidase(HRP)and a fluorescence substrate has been attracting great interests in developing sensitive biochemical analysis and immunoassays.10-Acetyl-3,7-dihydroxyphenoxazine(ADHP or A...The combination of horseradish peroxidase(HRP)and a fluorescence substrate has been attracting great interests in developing sensitive biochemical analysis and immunoassays.10-Acetyl-3,7-dihydroxyphenoxazine(ADHP or Amplex red)is the most sensitive fluorogenic substrate known for HRP in current market,however,it suffers from some drawbacks,such as non-specific reactivity to carboxylesterase and limited fluorescence stability.In the present study,a novel HRP substrate10-cyclopropylcarbonyl-dichloro-dihydroxyphenoxazine(AR-2),has been prepared,which exhibited improved sensitivity than ADHP in sensing HRP.Moreover,the fluorescence of AR-2/HRP demonstrated improved tolerance to physiological relevant p H fluctuation as compared to ADHP/HRP.Successful detection of uric acid/urate oxidase reaction indicated excellent application prospect of AR-2/HRP for monitoring H_(2)O_(2)-generating biochemical reactions.More interestingly,an enzyme-linked immunosorbent assay(ELISA)using AR-2 as the fluorescence reporter has been successfully used in detecting IgG against severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)from human serum samples.Overall,AR-2 exhibits improved performances over the commercial ADHP,which will be an ideal alternative to ADHP in HRP-based fluorescence biochemical analysis and immunoassays.展开更多
基金supported by the National Natural Science Foundation of China(Grant Nos.32571739,32270457,31872216,and 31670392)In addition,this study received approval from the Experimental Animal Ethics and Management Committee of Anhui University(IACUC(AHU)-2024-050).
文摘Dear Editor,Amplexus,a reproductive behavior in which the male clasps the female dorsally,is a characteristic of amphibians(Wells,2007).Most species exhibit either inguinal or axillary amplexus,although some reproduce without it(Wells,2007).While male-female amplexus is predominant,other forms such as male-male and multiple-male amplexus have also been documented(Soni et al.,2025).Release calls are typically produced by amplexed males to signal mating mismatches and mitigate the costs of inappropriate clasping(Kelehear and Shine,2019).Female-female amplexus,however,is rare and considered maladaptive,as it does not lead to fertilization(Chuang et al.,2019).
基金the funding support from Key-Area Research and Development Program of Guangdong Province(No.2022B1111020003)Guangzhou Talents Program for Innovation and Entrepreneurship(No.2021-L010)the Foshan“Blue Ocean Talent Program”for Innovation and Entrepreneurship(No.2230032002063)。
文摘The combination of horseradish peroxidase(HRP)and a fluorescence substrate has been attracting great interests in developing sensitive biochemical analysis and immunoassays.10-Acetyl-3,7-dihydroxyphenoxazine(ADHP or Amplex red)is the most sensitive fluorogenic substrate known for HRP in current market,however,it suffers from some drawbacks,such as non-specific reactivity to carboxylesterase and limited fluorescence stability.In the present study,a novel HRP substrate10-cyclopropylcarbonyl-dichloro-dihydroxyphenoxazine(AR-2),has been prepared,which exhibited improved sensitivity than ADHP in sensing HRP.Moreover,the fluorescence of AR-2/HRP demonstrated improved tolerance to physiological relevant p H fluctuation as compared to ADHP/HRP.Successful detection of uric acid/urate oxidase reaction indicated excellent application prospect of AR-2/HRP for monitoring H_(2)O_(2)-generating biochemical reactions.More interestingly,an enzyme-linked immunosorbent assay(ELISA)using AR-2 as the fluorescence reporter has been successfully used in detecting IgG against severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)from human serum samples.Overall,AR-2 exhibits improved performances over the commercial ADHP,which will be an ideal alternative to ADHP in HRP-based fluorescence biochemical analysis and immunoassays.