Uncoupling protein 1 (UCP1) expressed by the brown adipose tissue (BAT) in the mitochondrial crista acts as a homeostatic thermogenerator of eutherians. The evaluation of UCP1 expression in the BAT offers significant ...Uncoupling protein 1 (UCP1) expressed by the brown adipose tissue (BAT) in the mitochondrial crista acts as a homeostatic thermogenerator of eutherians. The evaluation of UCP1 expression in the BAT offers significant scientific insight, especially in studies targeting limited areas such as the periarterial and pericardial regions of small experimental mammals. However, the negligible amount of this adipose tissue would render the general quantitative evaluation of the protein unreliable because of lipid contamination and low protein concentration. To address this problem, we quantitatively evaluated UCP1 expression in the mitochondrion of the mouse interscapular BAT using immunoelectron microscopy and immunohistochemical studies using a combination of primary and secondary antibodies in scheme A (rabbit anti-UCP1 IgG/gold particle-conjugated goat anti-rabbit IgG), B (rabbit IgG/gold particle-conjugated goat anti-rabbit IgG), C (rabbit anti-UCP1 IgG/gold particle-unconjugated goat anti-rabbit IgG), and D (rabbit IgG/gold particle-unconjugated goat anti-rabbit IgG). Scheme A shows the immunopositive reaction of obvious gold particles in the mitochondrial area, whereas other procedures revealed less distinctive reactions. The distinctive gold particle immunoreaction comprised electrical high-density spots with a mean diameter of >5 nm. However, in scheme B, the electrical high-density spots were scattered outside the mitochondrion and were significantly smaller than 4 nm;schemes C and D demonstrated few immunoreactions. Logistic regression analysis between schemes A and B showed that the threshold diameter of the electrical high-density spots measuring >5 nm indicated a true positive immunoreaction to anti-UCP1 antibody specifically in the mitochondrial area. Minor statistical difference was observed in the primary anti-UCP1 antibody between polyclonal IgG and monoclonal antibodies. Therefore, immunoelectron microscopy might be useful for evaluating negligible protein expression in some limited areas, such as UCP1 expression in the BAT of small experimental animals.展开更多
【目的】研究分析无核白葡萄干贮期颜色变化。【方法】以吐鲁番无核白葡萄干为试材,用0.08 mm PE袋密封包装,贮藏于室温,0、5和15℃条件下,用纱布袋子包装后室温下存放作为CK,通过测定叶绿素含量、总酚含量、抗坏血酸氧化酶(AAO)活性、...【目的】研究分析无核白葡萄干贮期颜色变化。【方法】以吐鲁番无核白葡萄干为试材,用0.08 mm PE袋密封包装,贮藏于室温,0、5和15℃条件下,用纱布袋子包装后室温下存放作为CK,通过测定叶绿素含量、总酚含量、抗坏血酸氧化酶(AAO)活性、多酚氧化酶(PPO)活性的变化情况,探讨贮藏温度对无核白葡萄干颜色变化的影响。【结果】五个处理组的无核白葡萄干颜色都发生了不同程度的褐变,但0和5℃处理较其它处理组降低了褐变速率,减缓了叶绿素降解速度,较好的保持了其贮期的颜色;CK、室温和15℃处理贮期叶绿素降解速率较快,无核白葡萄干色泽明显的发黄或变褐。在贮藏150~300 d区间,叶绿素含量在CK与0和5℃处理之间有显著性差异,室温在0和5℃处理之间存在显著性差异,0与15℃之间也存在显著性差异。由各因素间的相关性得知,叶绿素含量与总酚含量、叶绿素含量与AAO活性之间,在5个处理组间都有极显著的相关性(P≤0.01);叶绿素含量与PPO活性在CK处理与室温处理间为显著的相关性(0.01<P≤0.05);0、5和15℃处理之间为极显著的相关性(P≤0.01)。总酚含量在CK处理与0、5℃间有显著性差异。总酚含量与AAO在五个处理组间都有极显著的相关性(P≤0.01);总酚含量与PPO活性在0、5和15℃处理之间都有极显著的相关性(P≤0.01)。贮期无核白葡萄干AAO活性与PPO活性受到贮藏温度的影响,0℃处理和5℃处理降低了贮期无核白葡萄干AAO活性和PPO活性,有助于减缓色泽的变化。在贮藏150~300 d区间,AAO活性在CK贮藏下与0和5℃处理间有显著性差异,室温与5℃处理间有显著性差异。【结论】贮藏温度对无核白葡萄干贮期褐变有一定影响,0和5℃处理可以较好的保持贮期无核白葡萄干的色泽。展开更多
文摘Uncoupling protein 1 (UCP1) expressed by the brown adipose tissue (BAT) in the mitochondrial crista acts as a homeostatic thermogenerator of eutherians. The evaluation of UCP1 expression in the BAT offers significant scientific insight, especially in studies targeting limited areas such as the periarterial and pericardial regions of small experimental mammals. However, the negligible amount of this adipose tissue would render the general quantitative evaluation of the protein unreliable because of lipid contamination and low protein concentration. To address this problem, we quantitatively evaluated UCP1 expression in the mitochondrion of the mouse interscapular BAT using immunoelectron microscopy and immunohistochemical studies using a combination of primary and secondary antibodies in scheme A (rabbit anti-UCP1 IgG/gold particle-conjugated goat anti-rabbit IgG), B (rabbit IgG/gold particle-conjugated goat anti-rabbit IgG), C (rabbit anti-UCP1 IgG/gold particle-unconjugated goat anti-rabbit IgG), and D (rabbit IgG/gold particle-unconjugated goat anti-rabbit IgG). Scheme A shows the immunopositive reaction of obvious gold particles in the mitochondrial area, whereas other procedures revealed less distinctive reactions. The distinctive gold particle immunoreaction comprised electrical high-density spots with a mean diameter of >5 nm. However, in scheme B, the electrical high-density spots were scattered outside the mitochondrion and were significantly smaller than 4 nm;schemes C and D demonstrated few immunoreactions. Logistic regression analysis between schemes A and B showed that the threshold diameter of the electrical high-density spots measuring >5 nm indicated a true positive immunoreaction to anti-UCP1 antibody specifically in the mitochondrial area. Minor statistical difference was observed in the primary anti-UCP1 antibody between polyclonal IgG and monoclonal antibodies. Therefore, immunoelectron microscopy might be useful for evaluating negligible protein expression in some limited areas, such as UCP1 expression in the BAT of small experimental animals.
文摘【目的】研究分析无核白葡萄干贮期颜色变化。【方法】以吐鲁番无核白葡萄干为试材,用0.08 mm PE袋密封包装,贮藏于室温,0、5和15℃条件下,用纱布袋子包装后室温下存放作为CK,通过测定叶绿素含量、总酚含量、抗坏血酸氧化酶(AAO)活性、多酚氧化酶(PPO)活性的变化情况,探讨贮藏温度对无核白葡萄干颜色变化的影响。【结果】五个处理组的无核白葡萄干颜色都发生了不同程度的褐变,但0和5℃处理较其它处理组降低了褐变速率,减缓了叶绿素降解速度,较好的保持了其贮期的颜色;CK、室温和15℃处理贮期叶绿素降解速率较快,无核白葡萄干色泽明显的发黄或变褐。在贮藏150~300 d区间,叶绿素含量在CK与0和5℃处理之间有显著性差异,室温在0和5℃处理之间存在显著性差异,0与15℃之间也存在显著性差异。由各因素间的相关性得知,叶绿素含量与总酚含量、叶绿素含量与AAO活性之间,在5个处理组间都有极显著的相关性(P≤0.01);叶绿素含量与PPO活性在CK处理与室温处理间为显著的相关性(0.01<P≤0.05);0、5和15℃处理之间为极显著的相关性(P≤0.01)。总酚含量在CK处理与0、5℃间有显著性差异。总酚含量与AAO在五个处理组间都有极显著的相关性(P≤0.01);总酚含量与PPO活性在0、5和15℃处理之间都有极显著的相关性(P≤0.01)。贮期无核白葡萄干AAO活性与PPO活性受到贮藏温度的影响,0℃处理和5℃处理降低了贮期无核白葡萄干AAO活性和PPO活性,有助于减缓色泽的变化。在贮藏150~300 d区间,AAO活性在CK贮藏下与0和5℃处理间有显著性差异,室温与5℃处理间有显著性差异。【结论】贮藏温度对无核白葡萄干贮期褐变有一定影响,0和5℃处理可以较好的保持贮期无核白葡萄干的色泽。