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单细胞模型中用于臭氧生物指示的荧光分析。

Fluorescent analysis for bioindication of ozone on unicellular models.

作者信息

Roshchina Victoria V, Yashin V A, Kuchin A V

机构信息

Institute of Cell Biophysics RAS, Institutskaya str. 3, Pushchino, Moscow Region, 142290, Russia,

出版信息

J Fluoresc. 2015 May;25(3):595-601. doi: 10.1007/s10895-015-1540-2. Epub 2015 Mar 17.

Abstract

Unicellular model plant systems (vegetative microspores of horsetail Equisetum arvense and pollen of six plant species Corylus avellana, Dolichothele albescens Populus balsamifera, Salix caprea, Saintpaulia ionantha, Tulipa hybridum, on which autofluorescence and fluorescence after histochemical treatment studied, have been represented as bioindicators of ozone. It has found that low doses of ozone 0.005 or 0.008 μl/l did not affect or stimulate the autofluorescence of the samples with the ability to germinate in an artificial medium. In higher ozone concentrations (0.032 μl/l) either the decrease in the intensity of the emission or changing in the position of the maxima in the fluorescence spectrum (new 515-520 nm maximum characteristic for the green-and yellow area has appeared) were observed. In dose of 0.2 μl/l, higher than above the threshold of danger to human health, autofluorescence in all samples fell down to up to zero, and there was no the ability to germinate. In this case the formation of lipofuscin-like compounds fluoresced in blue with maxima from 440 to 485 nm was observed. Stress metabolites, known as neurotransmitters biogenic amines, were found in treated cells as determined on the characteristic fluorescence at 460-480 nm in the samples after a specific histochemical reactions for catecholamines (with glyoxylic acid) or for histamine (with o-phthalic aldehyde). Increased intensity of the emission under the treatment with ozone (total doses from 0.012 to 0.032 μl/l) was associated with an increase in the concentrations of catecholamines and histamine. The fluorescent analysis on undamaged cells-possible bioindicators of ozone can be useful in ecomonitoring for earlier warning about health hazardous concentrations of this compound in the air.

摘要

单细胞模式植物系统(问荆Equisetum arvense的营养小孢子以及六种植物榛属Corylus avellana、白绵毛柱Dolichothele albescens、香脂杨Populus balsamifera、皱叶柳Salix caprea、非洲紫罗兰Saintpaulia ionantha、郁金香Tulipa hybridum的花粉,研究了其自发荧光和组织化学处理后的荧光)已被用作臭氧的生物指示物。研究发现,低剂量的臭氧(0.005或0.008微升/升)对能够在人工培养基中萌发的样品的自发荧光没有影响或有刺激作用。在较高的臭氧浓度(0.032微升/升)下,观察到发射强度降低或荧光光谱中最大值位置发生变化(出现了新的515 - 520纳米最大值,这是绿色和黄色区域的特征)。在0.2微升/升的剂量下,高于对人体健康的危险阈值,所有样品的自发荧光降至零,且失去了萌发能力。在这种情况下,观察到形成了在440至485纳米处有最大值的蓝色荧光脂褐素样化合物。在经过针对儿茶酚胺(与乙醛酸)或组胺(与邻苯二甲醛)的特定组织化学反应后,根据样品在460 - 480纳米处的特征荧光确定,在处理过的细胞中发现了作为神经递质的生物胺类应激代谢产物。用臭氧处理(总剂量为0.012至0.032微升/升)时发射强度增加与儿茶酚胺和组胺浓度增加有关。对未受损细胞进行荧光分析——可能作为臭氧的生物指示物——可用于生态监测,以便对空气中该化合物的健康危害浓度进行早期预警。

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