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Lili Chen Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing 400715, China

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Xiaolong Jiang Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing 400715, China

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Haiwei Feng Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing 400715, China

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Hongjuan Shi Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing 400715, China

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Lina Sun Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing 400715, China

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Wenjing Tao Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing 400715, China

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Qingping Xie Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing 400715, China

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Deshou Wang Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing 400715, China

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Estrogen, which is synthesized earlier in females than androgen in males, is critical for sex determination in non-mammalian vertebrates. However, it remains unknown that what would happen to the gonadal phenotype if estrogen and androgen were administrated simultaneously. In this study, XY and XX tilapia fry were treated with the same dose of 17α-methyltestosterone (MT) and 17β-estradiol (E2) alone and in combination from 0 to 30 days after hatching. Treatment of XY fish with E2 resulted in male to female sex reversal, while treatment of XX fish with MT resulted in female to male sex reversal. In contrast, simultaneous treatment of XX and XY fish with MT and E2 resulted in female, but with cyp11b2 and cyp19a1a co-expressed in the ovary. Serum 11-ketotestosteron level of the MT and E2 simultaneously treated XX and XY female was similar to that of the XY control, while serum E2 level of these two groups was similar to that of the XX control. Transcriptomic cluster analysis revealed that the MT and E2 treated XX and XY gonads clustered into the same branch with the XX control. However a small fraction of genes, which showed disordered expression, may be associated with stress response. These results demonstrated that estrogen could maintain the female phenotype of XX fish and feminize XY fish even in the presence of androgen. Simultaneous treatment with estrogen and androgen up-regulated the endogenous estrogen and androgen synthesis, and resulted in disordered gene expression and endocrine disruption in tilapia.

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Wenjing Wang Islet and Cell Processing Laboratory, Puget Sound Blood Center/Northwest Tissue Center, Seattle, Washington, USA
Department of Medicine, University of Washington School of Medicine, Seattle, Washington, USA
Department of Surgery and Department of Orthopedics and Sports Medicine, University of Washington School of Medicine, Seattle, Washington, USA
Pacific Northwest Research Institute, Seattle, Washington, USA

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Lisa Upshaw Islet and Cell Processing Laboratory, Puget Sound Blood Center/Northwest Tissue Center, Seattle, Washington, USA
Department of Medicine, University of Washington School of Medicine, Seattle, Washington, USA
Department of Surgery and Department of Orthopedics and Sports Medicine, University of Washington School of Medicine, Seattle, Washington, USA
Pacific Northwest Research Institute, Seattle, Washington, USA

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D Michael Strong Islet and Cell Processing Laboratory, Puget Sound Blood Center/Northwest Tissue Center, Seattle, Washington, USA
Department of Medicine, University of Washington School of Medicine, Seattle, Washington, USA
Department of Surgery and Department of Orthopedics and Sports Medicine, University of Washington School of Medicine, Seattle, Washington, USA
Pacific Northwest Research Institute, Seattle, Washington, USA

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R Paul Robertson Islet and Cell Processing Laboratory, Puget Sound Blood Center/Northwest Tissue Center, Seattle, Washington, USA
Department of Medicine, University of Washington School of Medicine, Seattle, Washington, USA
Department of Surgery and Department of Orthopedics and Sports Medicine, University of Washington School of Medicine, Seattle, Washington, USA
Pacific Northwest Research Institute, Seattle, Washington, USA

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JoAnna Reems Islet and Cell Processing Laboratory, Puget Sound Blood Center/Northwest Tissue Center, Seattle, Washington, USA
Department of Medicine, University of Washington School of Medicine, Seattle, Washington, USA
Department of Surgery and Department of Orthopedics and Sports Medicine, University of Washington School of Medicine, Seattle, Washington, USA
Pacific Northwest Research Institute, Seattle, Washington, USA

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In this study, we investigated the use of a novel oxygen biosensor system to detect changes in oxygen consumption rates (OCRs) by islets in response to glucose. Islets from non-human primate and human pancreata were seeded into an oxygen biosensor system microplate and exposed to basal (2.8 or 5.6 mM) or high (16.7 or 33.3 mM) glucose over either a long-term or a short-term culture. Our data clearly demonstrated that non-human primate islets cultured in high glucose conditions exhibited significant increases in OCRs over a 168 h extended culture period (P<0.05), which indicates an accelerated rate of β-cell metabolism triggered by glucose over time. Significant increases in OCRs (P<0.01) were also attained in both non-human primate and human islets exposed to high glucose conditions in a 120 min short-term incubation period. OCRs exhibited by human islets exposed to different glucose concentrations correlated with insulin secretion (r 2=0.7681, P<0.01). Moreover, the OCR stimulation index (i.e. OCR at high glucose/OCR at basal glucose) was significantly greater in human islets displaying high viabilities as opposed to islets exhibiting low viabilities (P<0.05). Together these data demonstrate that this novel oxygen biosensor system documents significant increases in islet oxygen consumption upon acute and chronic exposure to high glucose concentrations. Importantly, this methodology rapidly and robustly detects changes in OCRs by islets in response to high glucose stimulation that correlate well with the metabolic activities and functional viability of islets and clearly delineates significant differences in OCR stimulation index between high and low viability human islets, and therefore may prove to be an effective approach for quickly assessing the functional viability of islets prior to transplantation.

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Yu Wang Department of General Surgery, Jintan Affiliated Hospital of Jiangsu University, Jintan, Jiangsu, China

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Airong Wu Department of Gastroenterology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China

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Liting Xi Department of Gastroenterology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China

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Ji Yang Department of Geriatrics, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang, China

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Wenjing Zhou Department of Geriatrics, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang, China

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Yuming Wang Department of Geriatrics, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang, China

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Shuang Liang Medical School of Nantong University, Nantong, Jiangsu, China

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Weixin Yu Department of General Surgery, Jintan Affiliated Hospital of Jiangsu University, Jintan, Jiangsu, China

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Yue Wang Department of Hepatology, Suzhou No. 5 People’s Hospital, Suzhou, Jiangsu, China

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Jinzhou Zhu Department of Gastroenterology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China

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