蚯蚓发光体系及其应用研究

发光蚯蚓在世界各地普遍存在。Wampler研究了蚯蚓发光的生理学和生物化学性质。Mulkerrin等还分离纯化了发光蚯蚓的荧光酶,人工合成了荧光素。由于蚯蚓发光体系可用于许多生化分析,因而引起许多学者的注意和兴趣。本文首次报道从我国无锡发现的发光蚯蚓,并从其体液中提取荧光素酶粗提取物,在体外重新组成新的发光体系。研究了这一发光体系的性质,并对蚯蚓发光反应机制和可能应用作了初步探讨。     

对发光小蚯蚓的观察及研究

1987年1月,在无锡县华庄镇发现了一种可以发光的小蚯蚓(属于巨蚓科、棘蚓亚科),现将对其发光现象的研究结果报道如下: 发光小蚯蚓的生态环境发光小蚯蚓生活在阳暗、潮湿和腐殖质含量极为丰富的土壤中.我们是在一个厨房旁边     

萤火虫发光的化学机制

在若干昆虫中,普遍存在着生物冷光(Bioluminescence)现象。这些发光的昆虫大多数属于鞘翅目、萤科,通常称做萤火虫。这些昆虫发光有专门的复器,也是一种交配的信号。关于萤火虫发光的化学机理问题,不少学者进行过研究探讨。1947年,McElroy最早用北美洲萤火虫photinus pyralis作材料进行研究,发现萤火虫生物冷光要求ATP。1952年,B.L.斯特勒和J.R.托特尔用萤火虫生物冷光探测ATP,其灵敏度可达10~(-15)M(ATP);同年,Harvey研究发现,萤火虫发冷光的颜色是不同的,这是因为不同种的萤火虫含有不同类型的萤光素/萤光素酶系统。以后,M.德卢卡又进行了一系列的研究,提出了萤火虫发光反应的现代知识,这个被概述在图1中。     

生物中的超微弱发光

超微弱发光是所有生物都具有的一种普遍现象,发光强度极其微弱,量子效率也很低,波长范围广,自１９２３年发现超微弱发光以来,各国科学家做了大量的研究,对机理进行探讨,目眼微弱发光的柚是依然不很清楚。生物超微弱发光在许多领域得到广泛的应用,由超微弱发光研究发展为的发光检测技术快速,简便,而且不会对组织材料造成损伤。本语文就生物超微弱发光机理及应用研究进行了综述。     

漫谈生物的发光

发光生物的分布具有发光能力的生物,在大自然中分布很广,其中少数为一些细菌和真菌,大多数是存在于动物界的一些物种。细菌中的发光细菌(luminous bacteria),主要是海产的发光性水生微生物(plankton),目前已知的发光细菌,不下百十种之多,其中具有显著发光现象的有:磷光假单胞菌(photobacterium phosphoreum)、费氏无色菌(A chromobacter fisheri)、磷光弧菌(Vibrio phosphorescens)和发光杆菌(Bacillus photogenus)。这些发光细菌,除了常常引起夜间海面发光外,还由于它们的孳生繁殖,成为被污染的死鱼、水产加工品、尸肉在黑暗中发光的原因。在缺少冷藏设备的过去年代,     

中国发光蚓一新种(后孔目:巨蚓科,棘蚓亚科)

1987年在江苏省无锡县采到小型发光蚯蚓,经鉴定,为一新种—无锡微蠕蚓(Microscolex wuxiensis)本种与燐微蠕蚓[(Microscolex phosphoreus(Ant.Dug,)]虽近似,但仍有明显不同。     

发光甲虫与生物荧光

生物荧光是活体生物自身可以发光的有趣生命现象。具有这一现象的生物存在于生物四界中,但目前关于这一现象的研究报道主要来自于昆虫,尤其是以萤火虫为代表的发光甲虫的研究。文章对发光甲虫的分类地位、生物荧光发生的原理、发光器官的类型、闪光的“开关”机制、生物荧光的生物学意义及其相关行为学研究进展等进行了详细介绍。此外,还简要提及了荧光生物及其荧光酶的应用。这对了解及探讨生物荧光现象、加强对中国的发光甲虫及其它发光生物的研究及保护利用具有一定的借鉴作用。     

生物超弱发光及其应用研究概述

生物超弱发光是生物物理中光生物学的重要内容之一,自１９２３年以来,人们已进行了大量研究。本文评述了生物超弱发光的机理、测量和理化影响因素,总结了生物超弱发光在我国农业和医学中的应用研究,并对初步展望了生物超弱发光的未来研究方向。     

细菌荧光素酶体外发光体系的建立及应用于NADH定量检测

摘要：【目的】本研究旨在建立鳆发光杆菌（Photobacterium leiognathi）YL 荧光素酶：FMN-NADH氧化还原酶体外发光双酶体系,并对荧光素酶：FMN-NADH氧化还原酶体外发光双酶体系应用于NADH的定量检测进行初步探索。【方法】利用从鳆发光杆菌提取并经部分纯化的荧光素酶和FMN-NADH氧化还原酶,通过优化体系中各底物的添加量,实现荧光素酶的体外发光。【结果】荧光素酶：FMN-NADH氧化还原酶体外发光双酶体系为：1 mL酶液中添加100 μL十二烷醛（27 mmol/L）、0.5 μL FMN-Na（10 mmol/L）、300 μL NADH（0.14 mmol/L）。NADH与荧光素酶：FMN-NADH氧化还原酶体系的发光强度呈良好的线性关系,其线性范围为1.0×10-10 ～1.0×10-8 mol/L。【结论】荧光素酶：FMN-NADH氧化还原酶体外发光双酶体系可以简便、灵敏、快速的定量检测NADH,为其进一步应用于环境检测、食品卫生与安全等领域活细菌数量的检测奠定了基础。     

生物超弱发光研究

生物超弱发光是自然界普遍在的一种生命现象。本文概述了近年来生物超弱发光研究的进展,探讨了生物超弱发光在医学和农业上应用的可能性,并提出了该领域有待深入研究的问题。     

Luminometer development in the last four decades: recollections of two entrepreneurs

This article, written by two entrepreneurs in luminescence, traces their involvement in the major part of the interconnected innovation and development of luminometers, adenosine triphosphate (ATP) bioluminescence and other technologies from the mid‐1970s to 2011 that ushered in much of the field of luminometry as we know it today. Key developments leading to current commercial applications of ATP bioluminescence, luminescence immunoassay, cellular luminescence, reporter gene and other applications are described from the first tube luminometers derived from early luminescence studies using liquid scintillation counting technology to measuring bioluminescence from crude ATP and firefly tail extracts. Copyright © 2012 John Wiley & Sons, Ltd.     

Effect of concentrating and exposing the bioluminescent bacteria to the non‐luminescent allo‐bacterial extracellular products on their luminescence

Bioluminescence is a biochemical process occurring in many organisms. Bacterial bioluminescence has been investigated extensively that lead to many applications of such knowledge. Quorum sensing in the bioluminescent bacteria is a chemical signal process to recognize the strength of its own population to start luminescence in harmony. There is a mechanism in these bacteria to also recognize inter‐species strength. When there is a higher number of these bacteria, the possibility and frequency of cell–cell physical contact will be high. In this study, the physical proximity was artificially enhanced between cells and the effect on luminescence in the concentrated cells in the normal culture medium and in the presence of other non‐bacterial cell‐free supernatants was investigated. The role of such physical contact in the quorum sensing in the bioluminescence is not known. Increase in the luminescence of V. fischeri when concentrated shows that the presence of physical proximity facilitates the quorum sensing for their bioluminescence. Copyright © 2009 John Wiley & Sons, Ltd.     

A combination of NADHP and hispidin is not essential for bioluminescence in luminous fungal living gills of Mycena chlorophos

The chemical mechanisms underlying visible bioluminescence in the fungus Mycena chlorophos are not clear. A combination of dihydronicotinamide adenine dinucleotide phosphate (NADPH) and hispidin, which has been reported to increase the intensity of in vitro luminescence in crude cold‐water extracts prepared from the bioluminescent fruiting bodies of M. chlorophos, exhibited potential bioluminescence activation in the early bioluminescence stages, in which the bioluminescence was ultra‐weak, for living gills and luminescence activation for non‐bioluminescent gills, which was collapsed by freezing and subsequent thawing, at all bioluminescence stages. These abilities were not evident in considerably bioluminescent gills. These abilities were blocked by trans‐4‐hydroxycinnamic acid and trans‐3,4‐dihydroxycinnamic acid, which were identified as in vivo bioluminescence‐activating components. Original bioluminescence and bioluminescence produced from the addition of trans‐4‐hydroxycinnamic acid and trans‐3,4‐dihydroxycinnamic acid in living gills were almost completely inhibited by 10 mM NaN₃, whereas the luminescence produced form the combination of NADPH and hispidin in thawed non‐bioluminescent and living gills at the early weak bioluminescence stages was not inhibited by 10 mM NaN₃. Thus, the combination of NADPH and hispidin plays different roles in luminescence systems compared with essential bioluminescence systems, and the combination of NADPH and hispidin was not essential for visible bioluminescence in living gills.     

成熟小麦抗穗发芽能力与超弱发光关系的研究

生物体的超弱发光表现与其本身的生理活动密切相关,利用超弱发光为指标,测定和比较了小麦不同品种在成熟时抗穗发芽的能力．为其进一步应用,解决农业实际问题提供方法和依据．     

Exogenous compounds in studying the mechanism of electron-excited state formation in bioluminescence

Using a series of exogenous fluorescent molecules as potential energy acceptors, the hypothesis on the activity of the upper electron-excited states in bioluminescence was tested. The results in bacterial and firefly bioluminescent enzyme systems were compared. Similar activity to the energetic precursor in bacterial bioluminescence was not proven in the case of the firefly system, the result of a very efficient intramolecular energy transfer in the emitter of the firefly bioluminescence. The influence of a number of metallic salts on a bacterial bioluminescent enzyme system was studied. Bioluminescence inhibition coefficients were compared to the free energies of electron withdrawing of cations. The correlation shows that inhibition and activation of luminescence intensity result from the effects of cations on electron transfer in the bioluminescent system.     

人体体表超弱光发射的测量

研制了一种能在室温下正常工作的高灵敏度并具有极低暗计数的光子计数器。用它来研究人体体表超弱光的发射。测量结果表明：人体不同部位体表发光强度是很不一致的：手指发光最强、掌心次之、然后是面颊、前额、小臂、上臂、胸腹部等依次减弱。先照对体表的发光有很大影响。体表不同部位因光照引起发光强度的增加值差异很大;避光后先强的衰减速度也不同。体表的发光强度随体表温度的升高而增强。某些物质接触体表可以使体表发光强度发生很大变化。     

A new rapid and sensitive bioluminescence assay for antibiotics that inhibit protein synthesis

N aveh , A., P otasman , I., B assan , H. & U litzur , S. 1984. A new rapid and sensitive bioluminescence assay for antibiotics that inhibit protein synthesis. Journal of Applied Bacteriology 56 , 457–463.
A new sensitive, rapid and simple bioluminescence assay for antibiotics inhibiting protein synthesis is described. In this assay the ability of the tested antibiotic to inhibit the de novo synthesis of the enzymes participating in the bacterial luminescence system is determined by means of a dark variant of a luminous bacterium that undergoes prompt induction of the luminescence system with certain DNA-intercalating agents. Upon induction, the in vivo luminescence of the dark variant is increased more than 50-fold within 30 min. Antibiotics that block the de novo synthesis of protein limit the development of luminescence at a level that was found to be a function of the antibiotic concentration. The minimum detectable concentration of antibiotics in the bioluminescence test, after 45–60 min of incubation, was 0.1 μg ml for streptomycin, gentamicin, kanamycin, lincomycin and chlorampheni-col and 0.3 μg/ml for neomycin, clindamycin and spectinomycin. The new bioluminescence test has been used to assay these antibiotics in serum.     

Intracellular redox equilibrium and growth phase affect the performance of luciferase-based biosensors

Light emission from the bacterial luciferase operon has been variously exploited during last two decades. The use of convenient inducible promoters has granted significant degrees of specificity to whole cell-based assays for high-throughput screening and environmental monitoring. Nevertheless, unexplained unspecific responses have been repeatedly reported. Here, we show that the impairment of the intracellular biochemical equilibrium interferes with the luminescence produced by Escherichia coli and Staphylococcus aureus strains carrying the lux operon under constitutive or inducible control. Compounds as trimethoprim and methotrexate, by indirectly inducing NADPH accumulation, enhance light emission. Conversely, molecules driving the cell toward an oxidized state, as dimethyl sulfoxide, inhibit luminescence. These findings fit into the accepted biochemical pathway for bioluminescence, where NADPH and reducing equivalents are necessary for the production of luciferase substrates, although they do not directly take part into the light-emitting reaction. Moreover, we investigated the influence of induction timing upon the bioluminescence response from inducible reporter systems and demonstrated a correlation between the emitted light and the growth phase at which induction is performed. Our results provide explanations for some unspecific responses recorded so far in whole cell-based luminescent biosensors and emphasize the intrinsic limitations of this kind of reporting system.     

旧石器和古人类遗址释光测年技术的可靠性和测年上限

释光测年技术已成为旧石器和古人类遗址,尤其是现代人类遗址,建立年代框架的重要工具之一。这一技术提供了现代人类出现在非洲、亚洲和澳大利亚的最早年代证据。本文简要介绍了释光测年的基本原理,对释光测年的可靠性和上限及所受的影响因素进行了综述。光释光测年的精密度（相对标准误差σ）一般为5%-10%,在理想条件下σ<5%,但是σ>10%的情况也不少见。与大量其他测年方法所获结果的一致性表明,光释光测年技术是可靠的。光释光测年的上限与样品的释光性质及环境剂量率有关,释光可靠年龄最大可达1百万年。对大多数遗址50万年的测年上限是可行的,这个年代范围涵盖了所有的现代人遗址。不同样品或颗粒间的释光性质差异很大,因而它们有不同的测年上限。同一样品中钾长石比石英有更高的测年上限,同一矿物中不同的释光信号对应的测年上限也不同。     

Bioluminescence in oceanology

For analytical purposes bioluminescence can be used in three main ways: 1. luminescence measurement of bioluminescent system components isolated in vitro; 2. determination of luminous organisms' reaction to the in vivo test-action; 3. measurement of bioluminescence in marine ecological systems. The majority of the reports of this Symposium are dealing with the first two topics. The aim of our presentation is to draw attention to the third one. The possibilities of bioluminescent analysis are wider than its traditional scheme of applications in the laboratory, when the emitting system is withdrawn from a native source and is placed in a cuvette of the light measuring device. The reverse scheme is also possible, i.e. the device can be introduced into light emitting system such as a marine biocenosis–the community of the sea inhabitants–where we obtain a highly sensitive and rapid means of gaining the information on the vital activity of marine ecosystems, i.e. their spatial structure, rhythms, man's influence upon them, etc. The present communication will consider the possibilities of this form of bioluminescent analysis.