A series of modified chlorophylls, namely, pyrochlorophyll a, Zn-pheophytin a, Zn-pheophorbide a, chlorophyllide a, [3-acetyl]-chlorophyll a, and bacteriochlorophyll a, have been investigated in micellar solutions. The study is aimed at establishing the role played by the different functional groups of the chlorophyll in the molecular organization of chlorophylls in a microheterogeneous environment. The surfactants (AOT, CTAB, and Triton X-100) have been chosen mainly on the basis of the different charges carried by their polar heads, to study the effect of a point charge on the spectral characteristics of the pigments. Besides optical techniques used to investigate the spectral properties of the pigments, the state of the micellar system was studied by resonance light scattering (RLS) and NMR (self-diffusion and relaxation time measurements). The results of the UV-vis measurements evidence the role played by the functional groups of the chlorophylls in the formation of the different species in solution. In view of the few cases reported in the literature on blue-shifted chlorophyll species, special attention has been devoted, to the behavior of the modified chlorophylls in CTAB where a species absorbing around 642 nm is always formed. CD, RLS, and NMR data identify this species as a pigment-surfactant aggregate, in which the positive charge of the surfactant interacts with the C-13 2 ketoester group of a monomeric chlorophyll.
The phospholipid composition of Rhodobacter sphaeroides cells resuspended in various hypertonic solutions has been examined by thin-layer chromatography and ESI mass spectrometry. R. sphaeroides responds to hyperosmotic stress by increasing the amount of cardiolipin in the membranes; this phenomenon occurs in spheroplasts also. Cardiolipin increases quickly and continuously during the time when the cells are resuspended in hypertonic medium. The optimum of stimulation of the neosynthesis of cardiolipin during osmotic stress was found to be at external 1 osm. ESI-MS analyses allowed the identification of two different cardiolipins in R. sphaeroides: the tetravaccenylcardiolipin ([M - H](-), m/z 1456.9) and the trivaccenylmonopalmitoylcardiolipin ([M - H](-), m/z 1430.0).
Cyclodextrins, cyclic oligosaccharides composed of amylose subunits, are known to interact with mycotoxins. The interactions may be useful to analytical chemists by altering the properties of the mycotoxin of interest, namely the chromatographic properties, electrophoretic properties, fluorescence, or absorption of these fungal metabolites. Practical applications of these effects have been the incorporation of cyclodextrins into high-performance liquid chromatography and capillary electrophoresis methods for mycotoxin detection. Specific mycotoxins include those with a native fluorescence such as the aflatoxins, ochratoxin A (OTA) and zearalenone (ZEN) as well as those that can be rendered fluorescent through derivatization, such as T-2 toxin. The literature describing the applications of cyclodextrins in mycotoxin analysis is reviewed and an attempt to extend the use of cyclodextrins to the detection of labelled T-2 toxin is presented. Twenty cyclodextrins were evaluated for their ability to enhance the fluorescence emission of T-2 toxin derivatized with pyrene-1-carbonyl cyanide (T2-Pyr). This evaluation revealed that heptakis (2,6-di-O-methyl)--cyclodextrin (DIMEB), in particular, enhanced T2-Pyr fluorescence. DIMEB was used as a buffer modifier in a capillary electrophoresis-laser-induced fluorescence (CE-LIF) method for detecting T-2 in maize. Because of the effects that certain cyclodextrins have, especially under aqueous conditions, they may make useful additives for a variety of mycotoxin analytical methods.
Aptamers are synthetic single-stranded DNA or RNA sequences that can fold into tertiary structures allowing them to interact with and bind to targets with high affinity and specificity. This paper describes the first selection and identification of DNA aptamers able to recognize the biogenic amine tyramine. To successfully isolate aptamers to this challenging small molecule target, the SELEX methodology was adapted by combining a systematic strategy to increase the selection stringency and monitor enrichment success. As the benefits of applying high-throughput sequencing (HTS) in SELEX experiments is becoming more clear, this method was employed in combination with bioinformatics analysis to evaluate the utility of the selection strategy and to uncover new potential high affinity sequences. On the basis of the presence of consensus regions (sequence families) and family similarities (clusters), 15 putative aptamers to tyramine were identified. A recently described workflow approach to perform a primary screening and characterization of the aptamer candidates by microequilibrium dialysis and by microscale thermophoresis was next leveraged. These candidate aptamers exhibited dissociation constant (Kd) values in the range of 0.2-152 μM with aptamer Tyr_10 as the most promising one followed by aptamer Tyr_14. These aptamers could be used as promising molecular recognition tools for the development of inexpensive, robust and innovative biosensor platforms for the detection of tyramine in food and beverages.
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