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1.
Biosensors for rapid monitoring of primary-source drinking water using naturally occurring photosynthesis 总被引:4,自引:0,他引:4
Working with primary-source freshwater drinking samples from the Clinch and Tennessee Rivers, we have developed a tissue-based biosensor detection system that uses naturally occurring aquatic photosynthetic tissue as the sensing material for detection of chemical antagonists in the water. Sensor readout is based on well-known principles of fluorescence induction by living photosynthetic tissue. The Clinch River is the main source of drinking water for Oak Ridge, Tennessee, while the Tennessee River is a major source for the city of Knoxville. We have successfully detected algae in every sample that we examined and readily monitored changes in the characteristic fluorescence induction curves when the samples were exposed to potassium cyanide (KCN), methyl parathion (MPt), N'(3,4-dichlorophenyl)-N,N-dimethylurea (DCMU), and paraquat. The percentage decreases in photochemical yields observed in Tennessee River samples after a 24-min exposure to KCN, MPt, and DCMU were, respectively, 21.89+/-0.76, 3.28+/-0.18, and 14.77+/-1.81. For a site at the Clinch River, the percentage decreases were 22.78+/-1.63, 8.32+/-0.21, and 17.71+/-1.32 (Table 1). The unique aspect of this approach to real-time water quality monitoring is that unlike conventional sensing devices, this sensor material is external to the detecting instrument and is continuously refreshed. These biosensors may be used as continuous rapid-warning sentinels for detection of chemical warfare agents in sunlight-exposed drinking water supplies. 相似文献
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3.
Robin M. Black 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2010,878(17-18):1207-1215
This paper provides a short historical overview of the development of bioanalytical methods for chemical warfare (CW) agents and their biological markers of exposure, with a more detailed overview of methods for organophosphorus nerve agents. Bioanalytical methods for unchanged CW agents are used primarily for toxicokinetic/toxicodynamic studies. An important aspect of nerve agent toxicokinetics is the different biological activity and detoxification pathways for enantiomers. CW agents have a relatively short lifetime in the human body, and are hydrolysed, metabolised, or adducted to nucleophilic sites on macromolecules such as proteins and DNA. These provide biological markers of exposure. In the past two decades, metabolites, protein adducts of nerve agents, vesicants and phosgene, and DNA adducts of sulfur and nitrogen mustards, have been identified and characterized. Sensitive analytical methods have been developed for their detection, based mainly on mass spectrometry combined with gas or liquid chromatography. Biological markers for sarin, VX and sulfur mustard have been validated in cases of accidental and deliberate human exposures. The concern for terrorist use of CW agents has stimulated the development of higher throughput analytical methods in support of homeland security. 相似文献
4.
Carol-Visser J van der Schans M Fidder A Hulst AG van Baar BL Irth H Noort D 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2008,870(1):91-97
Rapid monitoring and retrospective verification are key issues in protection against and non-proliferation of chemical warfare agents (CWA). Such monitoring and verification are adequately accomplished by the analysis of persistent protein adducts of these agents. Liquid chromatography-mass spectrometry (LC-MS) is the tool of choice in the analysis of such protein adducts, but the overall experimental procedure is quite elaborate. Therefore, an automated on-line pepsin digestion-LC-MS configuration has been developed for the rapid determination of CWA protein adducts. The utility of this configuration is demonstrated by the analysis of specific adducts of sarin and sulfur mustard to human butyryl cholinesterase and human serum albumin, respectively. 相似文献
5.
Rizk SS Cuneo MJ Hellinga HW 《Protein science : a publication of the Protein Society》2006,15(7):1745-1751
The Escherichia coli phnD gene is hypothesized to code for the periplasmic binding component of a phosphonate uptake system. Here we report the characterization of the phosphonate-binding properties of the phnD protein product. We find that PhnD exhibits high affinity for 2-aminoethylphosphonate (5 nM), the most commonly occurring natural phosphonate produced by lower eukaryotes, but also binds several other phosphonates with micromolar affinities. A significant number of man-made phosphonates, such as insecticides and chemical warfare agents, are chemical threats and environmental pollutants. Consequently, there is an interest in developing methods for the detection and bioremediation of phosphonates. Bacterial periplasmic-binding proteins have been utilized for developing reagentless biosensors that report analytes by coupling ligand-binding events to changes in the emission properties of a covalently conjugated environmentally-sensitive fluorophore. Several PhnD conjugates described here show large changes in fluorescence upon binding to methylphosphonate (MP), with two conjugates exhibiting up to 50% decrease in emission intensity. Since MP is the final degradation product of many nerve agents, these PhnD conjugates can function as components in a biosensor system for chemical warfare agents. 相似文献
6.
A reduced model of the fluorescence from the cyanobacterial photosynthetic apparatus designed for the in situ detection of cyanobacteria 总被引:1,自引:0,他引:1
Beutler M Wiltshire KH Arp M Kruse J Reineke C Moldaenke C Hansen UP 《Biochimica et biophysica acta》2003,1604(1):33-46
Fluorometric determination of the chlorophyll (Chl) content of cyanobacteria is impeded by the unique structure of their photosynthetic apparatus, i.e., the phycobilisomes (PBSs) in the light-harvesting antennae. The problems are caused by the variations in the ratio of the pigment PC to Chl a resulting from adaptation to varying environmental conditions. In order to include cyanobacteria in fluorometric analysis of algae, a simplified energy distribution model describing energy pathways in the cyanobacterial photosynthetic apparatus was conceptualized. Two sets of mathematical equations were derived from this model and tested. Fluorescence of cyanobacteria was measured with a new fluorometer at seven excitation wavelength ranges and at three detection channels (650, 685 and 720 nm) in vivo. By employing a new fit procedure, we were able to correct for variations in the cyanobacterial fluorescence excitation spectra and to account for other phytoplankton signals. The effect of energy-state transitions on the PC fluorescence emission of PBSs was documented. The additional use of the PC fluorescence signal in combination with our recently developed mathematical approach for phytoplankton analysis based on Chl fluorescence spectroscopy allows a more detailed study of cyanobacteria and other phytoplankton in vivo and in situ. 相似文献
7.
The fast and slow kinetics of chlorophyll a fluorescence induction in plants, algae and cyanobacteria: a viewpoint 总被引:1,自引:0,他引:1
The light-induced/dark-reversible changes in the chlorophyll (Chl) a fluorescence of photosynthetic cells and membranes in the μs-to-several min time window (fluorescence induction, FI; or Kautsky transient) reflect quantum yield changes (quenching/de-quenching) as well as changes in the number of Chls a in photosystem II (PS II; state transitions). Both relate to excitation trapping in PS II and the ensuing photosynthetic
electron transport (PSET), and to secondary PSET effects, such as ion translocation across thylakoid membranes and filling
or depletion of post-PS II and post-PS I pools of metabolites. In addition, high actinic light doses may depress Chl a fluorescence irreversibly (photoinhibitory lowering; q(I)). FI has been studied quite extensively in plants an algae (less
so in cyanobacteria) as it affords a low resolution panoramic view of the photosynthesis process. Total FI comprises two transients,
a fast initial (OPS; for Origin, Peak, Steady state) and a second slower transient (SMT; for Steady state, Maximum, Terminal state), whose details are characteristically different in eukaryotic (plants and algae) and prokaryotic (cyanobacteria) oxygenic
photosynthetic organisms. In the former, maximal fluorescence output occurs at peak P, with peak M lying much lower or being
absent, in which case the PSMT phases are replaced by a monotonous PT fluorescence decay. In contrast, in phycobilisome (PBS)-containing
cyanobacteria maximal fluorescence occurs at M which lies much higher than peak P. It will be argued that this difference
is caused by a fluorescence lowering trend (state 1 → 2 transition) that dominates the FI pattern of plants and algae, and
correspondingly by a fluorescence increasing trend (state 2 → 1 transition) that dominates the FI of PBS-containing cyanobacteria.
Characteristically, however, the FI pattern of the PBS-minus cyanobacterium Acaryochloris marina resembles the FI patterns of algae and plants and not of the PBS-containing cyanobacteria. 相似文献
8.
Fluorometric determination of the chlorophyll (Chl) content of cyanobacteria is impeded by the unique structure of their photosynthetic apparatus, i.e., the phycobilisomes (PBSs) in the light-harvesting antennae. The problems are caused by the variations in the ratio of the pigment PC to Chl a resulting from adaptation to varying environmental conditions. In order to include cyanobacteria in fluorometric analysis of algae, a simplified energy distribution model describing energy pathways in the cyanobacterial photosynthetic apparatus was conceptualized. Two sets of mathematical equations were derived from this model and tested. Fluorescence of cyanobacteria was measured with a new fluorometer at seven excitation wavelength ranges and at three detection channels (650, 685 and 720 nm) in vivo. By employing a new fit procedure, we were able to correct for variations in the cyanobacterial fluorescence excitation spectra and to account for other phytoplankton signals. The effect of energy-state transitions on the PC fluorescence emission of PBSs was documented. The additional use of the PC fluorescence signal in combination with our recently developed mathematical approach for phytoplankton analysis based on Chl fluorescence spectroscopy allows a more detailed study of cyanobacteria and other phytoplankton in vivo and in situ. 相似文献
9.
Photosystem II-based biosensors for the detection of pollutants 总被引:1,自引:0,他引:1
Maria T. Giardi Michal Koblí zek Jirí Masojí dek 《Biosensors & bioelectronics》2001,16(9-12):1027-1033
Photosystem II (PSII) is the supramolecular pigment–protein complex in the chloroplast, which catalyses the light-induced transfer of electrons from water to plastoquinone (PQ) in a process that evolves oxygen. The PSII complex is also known to bind some groups of (photosynthetic) herbicides, heavy metals and other chemical substances that affect its activity. The objective of this study is to provide an overview of the systems available for the bioassay of pollutants using biosensors that are based on the photochemical activity of PSII. Some applications of the PSII-based biosensors including herbicide, heavy metal monitoring and the detection of radiation in space experiments are reported. 相似文献
10.
Matthew P. Willis Shawn M. Stevenson Thomas P. Pearl Brent A. Mantooth 《Journal of visualized experiments : JoVE》2014,(90)
The ability to directly characterize chemical transport and interactions that occur within a material (i.e., subsurface dynamics) is a vital component in understanding contaminant mass transport and the ability to decontaminate materials. If a material is contaminated, over time, the transport of highly toxic chemicals (such as chemical warfare agent species) out of the material can result in vapor exposure or transfer to the skin, which can result in percutaneous exposure to personnel who interact with the material. Due to the high toxicity of chemical warfare agents, the release of trace chemical quantities is of significant concern. Mapping subsurface concentration distribution and transport characteristics of absorbed agents enables exposure hazards to be assessed in untested conditions. Furthermore, these tools can be used to characterize subsurface reaction dynamics to ultimately design improved decontaminants or decontamination procedures. To achieve this goal, an inverse analysis mass transport modeling approach was developed that utilizes time-resolved mass spectroscopy measurements of vapor emission from contaminated paint coatings as the input parameter for calculation of subsurface concentration profiles. Details are provided on sample preparation, including contaminant and material handling, the application of mass spectrometry for the measurement of emitted contaminant vapor, and the implementation of inverse analysis using a physics-based diffusion model to determine transport properties of live chemical warfare agents including distilled mustard (HD) and the nerve agent VX. 相似文献
11.
Recent news from Syria on a possible use of chemical warfare agents made the headlines. Furthermore, the motivation of terrorists to cause maximal harm shifts these agents into the public focus. For incidents with mass casualties appropriate medical countermeasures must be available. At present, the most important threats arise from nerve agents and sulfur mustard. At first, self-protection and protection of medical units from contamination is of utmost importance. Volatile nerve agent exposure, e.g. sarin, results in fast development of cholinergic crisis. Immediate clinical diagnosis can be confirmed on-site by assessment of acetylcholinesterase activity. Treatment with autoinjectors that are filled with 2 mg atropine and an oxime (at present obidoxime, pralidoxime, TMB-4 or HI-6) are not effective against all nerve agents. A more aggressive atropinisation has to be considered and more effective oximes (if possible with a broad spectrum or a combination of different oximes) as well as alternative strategies to cope with high acetylcholine levels at synaptic sites should be developed. A further gap exists for the treatment of patients with sustained cholinergic crisis that has to be expected after exposure to persistent nerve agents, e.g. VX. The requirement for long-lasting artificial ventilation can be reduced with an oxime therapy that is optimized by using the cholinesterase status for guidance or by measures (e.g. scavengers) that are able to reduce the poison load substantially in the patients. 相似文献
12.
Martin H. Spalding Christa Critchley Govindjee William L. Orgren 《Photosynthesis research》1984,5(2):169-176
Carbon dioxide concentration during growth is commonly not considered to be a factor influencing the photochemical properties of plants. It was observed that fluorescence induction in Chlamydomonas reinhardii cells grown at air levels of CO2 was both qualitatively and quantitatively different from that of cells grown at 5% CO2. In the two cell types, measured at equivalent chlorophyll and irradiance levels, the fluorescence intensity and the ratio of the levels of peak fluorescence (Fp) to that of the initial fluorescence (Fo) were much lower in the air-adapted than in the 5% CO2 adapted cells. The maximum fluorescence (Fmax) in the presence of diuron was also lower for air-adapted cells. Roughly twice the light input was required for the air-adapted cells to give a fluorescence induction transient and intensity equivalent to that of the 5% CO2-adapted cells. Similar properties were observed in several other unicellular green algae and in cyanobacteria. Chlamydomonas grown under variable CO2 concentrations exhibit significant differences in photosynthetic carbon metabolism and are presumed to have altered energy requirements. The observed variation in fluorescence induction may be due to changes in the properties of the thylakoid reactions (e.g. cyclic electron flow) of Chlamydomonas cells, which may, in turn, be due to a response to the altered energy requirements. 相似文献
13.
Olsen EV Sorokulova IB Petrenko VA Chen IH Barbaree JM Vodyanoy VJ 《Biosensors & bioelectronics》2006,21(8):1434-1442
Proof-in-concept biosensors were prepared for the rapid detection of Salmonella typhimurium in solution, based on affinity-selected filamentous phage prepared as probes physically adsorbed to piezoelectric transducers. Quantitative deposition studies indicated that approximately 3 x 10(10)phage particles/cm(2) could be irreversibly adsorbed for 1 h at room temperature to prepare working biosensors. The quality of phage deposition was monitored by fluorescent microscopy. Specific-bacterial binding resulted in resonance frequency changes of prepared sensors, which were evaluated using linear regression analysis. Sensors possessed a rapid response time of <180 s, had a low-detection limit of 10(2)cells/ml and were linear over a range of 10(1)-10(7)cells/ml with a sensitivity of 10.9 Hz per order of magnitude of S. typhimurium concentration. Viscosity effects due to increasing bacterial concentration and non-specific binding were not significant to the piezoelectric platform as confirmed by dose-response analysis. Phage-bacterial binding was confirmed by fluorescence and scanning electron microscopy. Overall, phage may constitute effective bioreceptors for use with analytical platforms for detecting and monitoring bacterial agents, including use in food products and possibly biological warfare applications. 相似文献
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Photosynthetic fluorescence emission spectra measurement at the temperature of 77 K (–196°C) is an often-used technique in photosynthesis research. At low temperature, biochemical and physiological processes that modulate fluorescence are mostly abolished, and the fluorescence emission of both PSI and PSII become easily distinguishable. Here we briefly review the history of low-temperature chlorophyll fluorescence methods and the characteristics of the acquired emission spectra in oxygen-producing organisms. We discuss the contribution of different photosynthetic complexes and physiological processes to fluorescence emission at 77 K in cyanobacteria, green algae, heterokont algae, and plants. Furthermore, we describe practical aspects for obtaining and presenting 77 K fluorescence spectra. 相似文献
16.
Development of novel conductometric biosensors based on immobilised whole cell Chlorella vulgaris microalgae 总被引:2,自引:0,他引:2
A novel biosensor based on immobilised whole cell Chlorella vulgaris microalgae as a bioreceptor and interdigitated conductometric electrodes as a transducer has been developed and tested for alkaline phosphatase activity (APA) analysis. These sensors were also used for the detection of toxic compounds, namely cadmium ions, in aquatic habitats. Algae were immobilised inside bovine serum albumin (BSA) membranes cross-linked with glutaraldehyde vapours. The detection of the local conductivity variations caused by algae enzymatic reactions could be achieved. The inhibition of C. vulgaris microalgae Alkaline phosphatase activities in presence of cadmium ions was measured. These results were compared with measurements in bioassays. It finally appeared that conductometric biosensors using algae seemed more sensitive than bioassays to detect low levels of cadmium ions (the detection limit for the first experiments was 1 ppb of Cd2+). The main advantages of these alkaline phosphatase biosensors consist of their high specificity in regard to the toxic compounds they enable to detect, but also on their high stability since contrary to enzymatic biosensors, they use whole algae cells with APs on their walls. 相似文献
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Grimsley JK Calamini B Wild JR Mesecar AD 《Archives of biochemistry and biophysics》2005,442(2):169-179
Organophosphorus hydrolase detoxifies a broad range of organophosphate pesticides and the chemical warfare agents (CWAs) sarin and VX. Previously, rational genetic engineering produced OPH variants with 30-fold enhancements in the hydrolysis of CWA and their analogs. One interesting variant (H254R) in which the histidine at position 254 was changed to an arginine showed a 4-fold increase in the hydrolysis of demetonS (VX analog), a 14-fold decrease with paraoxon (an insecticide), and a 183-fold decrease with DFP (sarin analog). The three-dimensional structure of this enzyme at 1.9A resolution with the inhibitor, diethyl 4-methylbenzylphosphonate (EBP), revealed that the inhibitor did not bind at the active site, but bound exclusively into a well-defined surface pocket 12 A away from the active site. This structural feature was accompanied by non-competitive inhibition of paraoxon hydrolysis by EBP with H254R, in contrast to the native enzyme, which showed competitive inhibition. These parallel structure-function characteristics identify a functional, allosteric site on the surface of this enzyme. 相似文献
19.
Responses of aquatic algae and cyanobacteria to solar UV-B 总被引:4,自引:0,他引:4
Sinha Rajeshwar P. Klisch Manfred Gröniger Almut Häder Donat-P. 《Plant Ecology》2001,154(1-2):219-236
Continuous depletion of the stratospheric ozone layer has resulted in an increase in solar ultraviolet-B (UV-B; 280–315 nm) radiation reaching the Earth's surface. The consequences for aquatic phototrophic organisms of this small change in the solar spectrum are currently uncertain. UV radiation has been shown to adversely affect a number of photochemical and photobiological processes in a wide variety of aquatic organisms, such as cyanobacteria, phytoplankton and macroalgae. However, a number of photosynthetic organisms counteract the damaging effects of UV-B by synthesizing UV protective compounds such as mycosporine-like amino acids (MAAs) and the cyanobacterial sheath pigment, scytonemin. The aim of this contribution is to discuss the responses of algae and cyanobacteria to solar UV-B radiation and the role of photoprotective compounds in mitigating UV-B damage. 相似文献
20.
Fred M. Cowan Clarence A. Broomfield William J. Smith 《Cell biology and toxicology》1991,7(3):239-248
Sulfur mustard is a waemical warfare blistering agent for which neither the mechanism of action nor an antidote is known. Papirmeister et al. (1985) have postulated a biochemical hypothesis for mustard-induced cutaneous injury involving a sequelae of DNA alkylation, metabolic disruption and activation of protease. Human peripheral blood lymphocytes in cell cultures were employed as an in vitro model for alkylating agent toxicity. A chromogenic peptide substrate assay was used for detection of protease in lymphocytes treated with sulfur mustard or chloroethyl ethyl sulfide. Exposure of human peripheral blood lymphocytes from normal donors to these alkylating agents resulted in an increase in cell associated protease activity. This increase in protease activity may contribute to the pathology or act as an indicator to predict methods of therapeutic intervention for sulfur mustard toxicity.Abbreviations PBL
peripheral blood lymphocytes
- CEES
chloroethyl ethyl sulfide
- DFP
diisopropyl fluoro-phosphate
- pNA
p-nitroaniline
- CPSPA
Chromogenic Peptide Substrate Protease Assay
The opinions or assertions herein are the private views of the authors and are not to be construed as official or as reflecting the views of the Department of the Army or the Department of Defense. 相似文献