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Singlet oxygen production in herbicide-treated photosystem II.

Photo-generated reactive oxygen species in herbicide-treated photosystem II were investigated by spin-trapping. While the production of .OH and O2-* was herbicide-independent, 1O2 with a phenolic was twice that with a urea herbicide. This correlates with the reported influence of these herbicides on the redox properties of the semiquinone QA-* and fits with the hypothesis that 1O2 is produced by charge recombination reactions that are stimulated by herbicide binding and modulated by the nature of the herbicide. When phenolic herbicides are bound, charge recombination at the level of P+*Pheo-* is thermodynamically favoured forming a chlorophyll triplet and hence 1O2. With urea herbicides this pathway is less favourable.

Binding Sites↗

Direct detection of singlet oxygen via its phosphorescence from cellular and fungal cultures.

In summary, we have developed methods to detect 1O2 spectrally from viable keratinocytes and fungal mycelial cultures. Both time-resolved and steady-state spectrophotometers were used, providing complementary information on optimal conditions for the unambiguous detection of 1O2 phosphorescence. By using our techniques, we were able to confirm that photosensitizers in contact with living cells indeed generate 1O2. The model systems and the procedures described can be adopted for other studies involving 1O2 and oxidative stress in living cells.

Ascomycota↗

Fluorescence decay of singlet excited-state of safranine T and its interaction with ground-state of pyridinthiones in micelles and homogeneous media.

The fluorescence decays of safranine T were studied in different homogeneous solvents and heterogeneous micellar solutions. It has been found that micellization leads to an increase in the lifetime. The lifetime distributions were studied in micelles and homogeneous media. It was found that the different half-width distributions of the dye in different micelles are related to the different orientation of the dye in different micelles. Also, the fluorescence quenching of safranine T by 4,6-disubstituted-3-cyanopyridin-2(1H)-thiones was studied in chloroform, methanol and acetonitrile as well as in different micelles. In heterogeneous media the kq values for quenching of safranine T by thiones in various micelles increase on the following order: kq(CTAB) < kq(TX-100) < kq(SDS). This is due to the electrostatic interactions between the anionic SDS and the cationic moiety of safranine T and therefore the quenching process will be less significant.

Fluorescence↗

Phenylalanine-to-tyrosine singlet energy transfer in the archaebacterial histone-like protein HTa.

The Archaebacterium Thermoplasma acidophilum has a histone-like protein (HTa) abundantly associated with its deoxyribonucleic acid. Each native tetrameric complex of HTa contains 20 phenylalanine residues, 4 tyrosine residues, and no tryptophan. When the protein was excited by radiation at 252 nm, which is a wavelength absorbed predominantly by phenylalanine, the fluorescent emission was mostly from tyrosine. According to the excitation spectrum for this tyrosine fluorescence, the cause was energy transfer from phenylalanine, which occurred with about 50% efficiency. When the tyrosine residues were removed enzymatically, the excited-state lifetime of the phenylalanine residues nearly doubled. Because of energy transfer, the tyrosine emission had two apparent fluorescence decay lifetimes; one lifetime (3.9 ns) was that of tyrosine while the second (12.1 ns) corresponded to the excited state of phenylalanine.

Archaea↗