Determination of pH gradients in intact cyanobacteria by electron spin resonance spectroscopy.
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Cyanobacteria↗
Biomedical subjects
S Belkin
Publications and source records attributed to S Belkin.
Determination of dissolved oxygen in photosynthetic systems by nitroxide spin-probe broadening.
Concentrations of dissolved oxygen were monitored by following the width of the midfield line of the electron spin resonance spectrum of a nitroxide spin-probe. Measurements of peak-to-trough widths of first derivative spectra yielded accurate data over a high range of O2 concentrations (up to 5mM). Continuous traces of second harmonic line heights yielded similar results and proved to be advantageous for kinetic measurements, but were nonlinear and less sensitive at O2 levels above 2 mM. Photosynthetic oxygen evolution and respiratory oxygen uptake in the cyanobacterium Agmenellum quadruplicatum were thus examined over a broad range of oxygen concentrations. Upon prolonged (greater than 1 min) illumination, effects of photooxidative damage to both photosynthesis and respiration were demonstrated in the same experimental system. With the addition of an impermeable paramagnetic broadening agent, rapid transients in intracellular concentrations of dissolved O2 also could be measured.
Cyanobacteria↗
Reduction and destruction rates of nitroxide spin probes.
A series of nitroxides was tested for rates of one-electron reduction in a chemical, a photochemical, and two biological systems by ESR assays. In all cases, piperidine and hydropyridine nitroxides were reduced consistently more rapidly than pyrroline and pyrrolidine nitroxides. Substituents on the nitroxides also affected reduction rates, although not as greatly as ring structure. One of the reduction systems, consisting of the photosensitizer FMN and the photoreductant EDTA, was used to study both anaerobic reduction and O2-dependent reoxidation of some of the nitroxides. Reduced piperidine and hydropyridine nitroxides were also oxidized more rapidly than the reduced pyrroline and pyrrolidine nitroxides. Reoxidation subsequent to reduction was partially inhibited by superoxide dismutase, indicating that superoxide radicals are involved in the process. Even after prolonged reoxidation, not all of the probe molecules were returned to their oxidized form, implying an irreversible "destruction" of the spin probe concomitant with its chemical reduction. Probe destruction was studied more specifically with a photochemical system for generating methyl radicals, which showed that these carbon-centered radicals destroyed different nitroxides at rates which were much less influenced by the nitroxide structures than one-electron reduction was.
Animals↗
Proton gradients in intact cyanobacteria.
The internal pH values of two unicellular cyanobacterial strains were determined with electron spin resonance probes, over an external pH range of 6 to 9, in the light and in the dark. The slow growing, thylakoid-lacking Gloeobacter violaceus was found to have a low capacity for maintaining a constant internal pH. The distribution pattern of weak acid and amine nitroxide spin probes across the cell membranes of this organism, in the light and in the dark, was consistent with the assumption that it contains a single intracellular compartment. At an external pH of 7.0, intracellular pH was 6.8 in the dark and 7.2 in the light. The cells of Agmenellum quadruplicatum, a marine species, were found to contain two separate compartments; in the dark, the pH of the cytoplasmic and the intrathylakoid spaces were calculated to be 7.2 and 5.5, respectively. Upon illumination, the former increased and the latter decreased by about 0.5 pH units.
Biological Transport↗
A new sulfur-reducing, extremely thermophilic eubacterium from a submarine thermal vent.
A newly described bacterial isolate, designated strain NS-E, differs from presently known extremely thermophilic bacteria in various characteristics. It is a strictly heterotrophic eubacterium of marine origin and has a temperature range for growth of 50 to 95 degrees C with an optimum at 77 degrees C and a pH of 7.5. Its DNA base composition is 41.3 mol% guanine + cytosine. It is obligately anaerobic, utilizes various sugars as well as yeast extract, and reduces elemental sulfur facultatively to hydrogen sulfide. In 24-h cultures cell densities are up to fourfold higher in the presence than in the absence of elemental sulfur. Sulfide concentrations of 1.0 and 10.0 mM limit growth by 65 and 95%, respectively. Oxygen sensitivity is apparent only at or above that range of temperature at which growth occurs.
Journal Article↗
Biological and abiological sulfur reduction at high temperatures.
Reduction of elemental sulfur was studied in the presence and absencè of thermophilic sulfur-reducing bacteria, at temperatures ranging from 65 to 110 degrees C, in anoxic artificial seawater media. Above 80 degrees C, significant amounts of sulfide were produced abiologically at linear rates, presumably by the disproportionation of sulfur. These rates increased with increasing temperature and pH and were enhanced by yeast extract. In the same medium, the sulfur respiration of two recent thermophilic isolates, a eubacterium and an archaebacterium, resulted in sulfide production at exponential rates. Although not essential for growth, sulfur increased the cell yield in both strains up to fourfold. It is suggested that sulfur respiration is favored at high temperatures and that this process is not limited to archaebacteria, but is shared by other extreme thermophiles.
Journal Article↗
Na-Dithionite Promotes Photosynthetic Sulfide Utilization by the Cyanobacterium Oscillatoria limnetica.
The light- and sulfide-dependent induction process leading to photosystem I-mediated sulfide utilization by Oscillatoria limnetica, for either H(2) evolution or CO(2) photoassimilation, was studied. The identical dependence on pH of the lag length, the inhibition of leucine incorporation and final H(2)S concentration imply that the latter exerts a deleterious effect on nonadapted cells.Na-dithionite (Na(2)S(2)O(4)), Na-sulfite (Na(2)SO(3)), or ethanol cannot serve as photosynthetic electron donors. However, when these compounds were added to the sulfide-containing system, the need for induction was eliminated. At pH 6.9, in the presence of 3.5 millimolar sulfide, these substances (at concentrations of 10 millimolar, 5 millimolar, or 0.4 molar, respectively) completely abolished the delay preceding sulfide-dependent H(2) evolution. It is suggested that all three compounds expose a site capable of directly accepting sulfide electrons.Only dithionite could adapt the cells to sulfide utilization on its own. Sulfite or ethanol acted only in the presence of sulfide. It is implied that this specific activity of dithionite is related to its characteristic low redox potential.Sulfide-dependent H(2) evolution was insensitive to 3-(3,4-dichlorophenyl)-1,1-dimethylurea, but was inhibited by the plastoquinone antagonist 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone, in the presence as well as in the absence of dithionite. In both cases, therefore, the plastoquinone was implied in the electron transport from sulfide.
Journal Article↗
Hydrogen metabolism in the facultative anoxygenic cyanobacteria (blue-green algae) Oscillatoria limnetica and Aphanothece halophytica.
Two facultative anoxygenic photoautotrophic cyanobacteria, Oscillatoria limnetica and Aphanothece halophytica were found capable of CO2 photoassimilation using molecular hydrogen as electron donor in a photosystem I driven reaction. A. halophytica was also capable of evolving hydrogen from Na-dithionite reduced methylviologen in a light independent reaction.
Anaerobiosis↗
Birth defects involving the spine.
There is a multiplicity of birth defects of the spine occurring as isolated anomalies or as aspects of multifaceted syndromes. The clinical significance of these anomalies varies from the asymptomatic to the life threatening. Symptoms and signs may present at birth or not until years later. The optimal clinical result for the patient requires a careful history and physical examination, appropriate routine and specialized roentgenograms, laboratory data, and a high index of suspicion for signs or symptoms of neurological dysfunction. When the diagnosis is made, conservative or surgical treatment may be instituted as appropriate.
Atlanto-Occipital Joint↗
Water toxicity detection by a panel of stress-responsive luminescent bacteria.
A panel of Escherichia coli strains harbouring different stress-responsive promoters fused to a lux reporter system was used to assess the potential toxicity of 17 unknown model water samples. Using liquid cultures, nine out of 14 toxic samples were properly identified as toxic, whereas five were false negatives. All three non-toxic controls were identified correctly (no false positives). Two strains containing promoter-lux fusions were also tested when immobilized onto fibre-optic tips. One genotoxic sample and six toxic samples were correctly identified in this manner. The potential advantages and limitations in the use of genetically engineered bacteria as biosensors for water toxicity are discussed in view of these results.
Biosensing Techniques↗
Reporter gene bioassays in environmental analysis.
In parallel to the continuous development of increasingly more sophisticated physical and chemical analytical technologies for the detection of environmental pollutants, there is a progressively more urgent need also for bioassays which report not only on the presence of a chemical but also on its bioavailability and its biological effects. As a partial fulfillment of that need, there has been a rapid development of biosensors based on genetically engineered bacteria. Such microorganisms typically combine a promoter-operator, which acts as the sensing element, with reporter gene(s) coding for easily detectable proteins. These sensors have the ability to detect global parameters such as stress conditions, toxicity or DNA-damaging agents as well as specific organic and inorganic compounds. The systems described in this review, designed to detect different groups of target chemicals, vary greatly in their detection limits, specificity, response times and more. These variations reflect on their potential applicability which, for most of the constructs described, is presently rather limited. Nevertheless, present trends promise that additional improvements will make microbial biosensors an important tool for future environmental analysis.
Biological Assay↗
Microbial sensors of ultraviolet radiation based on recA'::lux fusions.
Escherichia coli strains containing plasmid-borne fusions of the recA promoter-operator region to the Vibrio fischeri lux genes were previously shown to increase their luminescence in the presence of DNA damage hazards, and thus to be useful for genotoxicant detection. The present study expands previous work by demonstrating and investigating the luminescent response of these strains to ultraviolet radiation. Several genetic variants of the basic recA'::lux design were examined, including a tolC modification of membrane efflux capacity, a chromosomal integration of the recA'::lux fusion, a different lux reporter (Photorhabdus luminescens instead of V. fischeri, allowing the assay to be run at 37 degrees C), and a different host bacterium (Salmonella typhimurium instead of E. coli). Generally, two modifications provided the fastest responses: the use of the S. typhimurium host or the P. luminescens lux reporter. Highest sensitivity, however, was demonstrated in an E. coli strain in which a single copy of the V. fischeri lux fusion was integrated into the bacterial chromosome.
Bacterial Proteins↗