Outstanding women scientists who have broadened the knowledge on biological photoreceptors-II.

This part II is a continuation of the article published in Photochemical and Photobiological Sciences (2023) 22, 2799-2815, https://doi.org/10.1007/s43630-023-00487-1 , which should be considered a work in progress. Now, two female scientists who have worked on different aspects of chronobiology, plus a younger colleague who recently and too prematurely died, are incorporated to the list of outstanding women who have expanded the knowledge in the field of biological photoreceptors.

Outstanding women scientists who have broadened the knowledge on biological photoreceptors.

Short biographical sketches are given of women born before 1955 who have contributed to our knowledge on the function, structure, and molecular basis of biological photoreceptors, both energy converters and photosensors.

Photophysics of Xanthene Dyes at High Concentrations in Solid Environments: Charge Transfer Assisted Triplet Formation.

The photophysical behavior of two xanthene dyes, Eosin Y and Phloxine B, included in microcrystalline cellulose particles is studied in a wide concentration range, with emphasis on the effect of dye concentration on fluorescence and triplet quantum yields. Absolute fluorescence quantum yields in the solid-state were determined by means of diffuse reflectance and steady-state fluorescence measurements, whereas absolute triplet quantum yields were obtained by laser-induced optoacoustic spectroscopy and their dependence on dye concentration was confirmed by diffuse reflectance laser flash photolysis and time-resolved phosphorescence measurements. When both quantum yields are corrected for reabsorption and reemission of radiation, ΦF values decrease strongly on increasing dye concentration, while a less pronounced decay is observed for ΦT . Fluorescence concentration quenching is attributed to the formation of dye aggregates or virtual traps resulting from molecular crowding. Dimeric traps are however able to generate triplet states. A mechanism based on the intermediacy of charge-transfer states is proposed and discussed. Calculation of parameters for photoinduced electron transfer between dye molecules within the traps evidences the feasibility of the proposed mechanism. Results demonstrate that photoactive energy traps, capable of yielding dye triplet states, can be formed even in highly-concentrated systems with random dye distributions.

Special issue dedicated to the memory of Elsa Beatriz Abuin Saccomano (1942-2012).

In memoriam of Elsa Abuin (1942-2012), teacher, mentor and friend. This Special Issue presents a collection of review articles and papers dedicated to the memory of Elsa Abuin and most of them presented during the Meeting of the Latin-American photochemists (Encuentro Latino-Americano de Fotoquímica y Fotobiología, ELAFOT) held in October 2012 in Córdoba, Argentina.

Triplet quantum yields in light-scattering powder samples measured by laser-induced optoacoustic spectroscopy (LIOAS).

In recent years, different methods and techniques have been applied to study the primary photophysical processes occurring in dye-loaded light-scattering powdered samples. In spite of this, there are still no reliable methods for the determination of triplet quantum yields for this kind of systems. Laser-induced optoacoustic spectroscopy (LIOAS) has been extensively used for the determination of triplet quantum yields of dyes in solution. In a previous work, LIOAS was applied to the measurement of absolute emission quantum yields of highly fluorescent powdered samples. Excellent agreement was found with values obtained from reflectance data. In this work, we apply the same technique for the determination of triplet quantum yields of Rose Bengal and Erythrosine B adsorbed on microcrystalline cellulose. In contrast to water and other solvents, internal conversion cannot be neglected in the cellulose environment. The triplet quantum yield for both dyes is around 0.55 and does not change with dye concentration.

Structural volume changes upon triplet formation of water-soluble porphyrins depend on the resonant effect of the substituents.

Laser-induced optoacoustic spectroscopy was applied to evaluate the photoinduced structural volume changes upon triplet state formation, Δ(T)V, of an aqueous solution of 5,10,15,20-tetrakis-(4-carboxyphenyl)-porphin. Two molecular forms differing in the ionization state of the carboxylic groups at the para-position of the phenyl ring were studied. The contractions, Δ(T)V, for all water-soluble 5,10,15,20-tetra-aryl-porphyrin compounds studied to date show a linear correlation with the Hammett resonant σ(R) constant. This indicates that the resonance electronic communication spreading over the π-orbitals between the meso-aryl rings and the porphyrin macrocycle determines the molecular contraction of the aqueous sphere around the macrocycle in the triplet state.

The history of ozone. Part VIII. Photochemical formation of ozone.

The historical development of the photochemical formation of ozone in the atmosphere is outlined, starting from the discovery of ozone by Schönbein in 1843 and the postulation of its role as UV filter by Hartley in 1881.

Quantum yield: the term and the symbol. A historical search.

Einstein's Photochemical Equivalence Law, i.e. one quantum of light absorbed per molecule, played a very important role in the development of photochemistry in the early part of the 20th century. The corresponding terminology, quantum yield and its symbol Phi, became accepted usage in a haphazard manner, which is presented in this paper.

Photophysical properties of structurally and electronically modified flavin derivatives determined by spectroscopy and theoretical calculations.

Four different riboflavin (RF) derivatives, two electronically modified compounds (1- and 5-deazariboflavin, 1DRF and 5DRF) and two sterically modified compounds (7,8-didemethyl- and 8-isopropylriboflavin, DMRF and iprRF), were subjected to a combination of time-resolved measurements (absorption and fluorescence) and high-level quantum chemical investigations. Both alkyl-modified flavins showed similar fluorescence properties as the parent compound, yet 5DRF had a larger quantum yield of fluorescence (PhiF = 0.52) than RF (PhiF = 0.27). Interestingly, 1DRF did not show fluorescence at all under these steady state conditions. The triplet quantum yield was different for the modified flavins such that no triplet formation was found for 1DRF, whereas the other compounds all formed triplet states (PhiTR for 5DRF of 0.64 and 0.50 and 0.23 for iprRF and DMRF, respectively). The triplet states of the two alkyl-modified flavins decayed with similar time constants as the parent compound, whereas a shorter lifetime was measured for 5DRF (tauTR = 15 micros, compared to tauTR = 29 micros for RF). In the calculations, the flavin derivatives were modeled as lumiflavins, that is, without the ribityl chain. We conclude that for aqueous solutions of DMRF, iprRF, and 5DRF intersystem crossing (ISC) takes place from the S1 1(pipi*) to the T2 3(pipi*) state by a vibronic spin-orbit coupling mechanism, a process common to most flavins, whereas ISC is slow in excited 1DRF due to the absence of a close-by triplet state.

Validation of fluorescence quantum yields for light-scattering powdered samples by laser-induced optoacoustic spectroscopy.

Determination of quantum yields for various processes (such as fluorescence and triplet formation) in dye-loaded light-scattering powdered samples is an open issue. Here, we report the testing of laser-induced optoacoustic spectroscopy (LIOAS) for the determination of fluorescence quantum yields of Rhodamine 101 and Rhodamine 6G adsorbed at various loadings on microgranular cellulose powder. The results of the LIOAS experiments are consistent with those from the method developed in one of our laboratories based on the measurement of apparent reflectance using an integrating sphere [Mirenda, M.; Lagorio, M. G.; San Roman, E. Langmuir 2004, 20, 3690-3697], which allows the simultaneous calculation of fluorescence quantum yield and reflectance devoid of fluorescence artifacts. Criteria to quantify overall errors and detect outlying values are developed. The theory underlying the application of LIOAS to light-scattering samples and experimental details are presented.

Flavodoxin relaxes in microseconds upon excitation of the flavin chromophore: detection of a UV-visible silent intermediate by laser photocalorimetry.

A small contraction concomitant with the relaxation of the protein in ca. 3 mus is observed upon ns-laser excitation at 455 nm of the Cys69Ala (C69A) mutant of flavodoxin II from Azotobacter vinelandii. This constitutes another example of detection of a UV-vis silent transient species through a photocalorimetric technique. The contraction is attributed to reorganization of protein dihedral bonds and of water molecules at relatively long distances from the flavin chromophore, after the protein has received the heat shock from the relaxing photoproduced charge transfer state. This study constitutes a preliminary step towards the understanding of the origin of protein movements taking place upon electron transfer reactions, e.g. between a photoinduced electron donor (or acceptor) and an accepting (or donating) cofactor in a protein.

Pitfalls and limitations in the practical use of Förster's theory of resonance energy transfer.

Disparate presentations in the literature of the basic equations of Förster's theory of resonance energy transfer are clarified and the limitations of these equations are discussed.

A photoprotection mechanism involving the D(2) branch in photosystem II cores with closed reaction centers.

Nanosecond transient absorption spectroscopy has been used to study reaction centre (RC) chlorophyll triplet quenching by carotenoid in intact photosystem II cores from T. elongatus with closed RCs. We found a triplet beta-carotene ((3)Car) signal (absorption difference maximum at 530 nm) that is sensitized by the RC chlorophyll (Chl) triplet with a formation time of ca. 190 ns, has a decay time of 7 micros and is formed with a quantum yield between 10 and 20%. The (3)Car signal is assigned to the beta-carotene on the D(2) branch of the RC. We thus propose a new photoprotection mechanism operative in closed RCs where-as a consequence of the negative charge on the quinone Q(A)-the triplet chlorophyll ((3)Chl) is formed by the radical pair (RP) mechanism on the normally inactive D(2) branch where it can be subsequently quenched by the D(2) beta-carotene. We suggest that the D(2) branch becomes active when the RCs are closed under high light fluence conditions. Under these conditions the D(2) branch plays a photoprotective role. This interpretation allows combining many seemingly inconsistent observations in the literature and reveals the so far missing RC triplet quenching mechanism in photosystem II. The newly proposed mechanism also explains the reason why this RC triplet quenching is not observed in isolated D(1)-D(2)-cyt b(559) RCs. If Q(A) is either not present at all (as in the isolated RC) or is not charged (as in open RCs or with doubly reduced Q(A)) then the RC (3)Chl is formed on the D(1) branch. The D(1) branch (3)Chl can not be quenched due to the large distance to the beta-carotene. This interpretation is actually in line with the well-known (3)RC quenching mechanism in bacterial RCs, where also the carotenoid in the (analogous to the D(2) branch) B-branch of the RC becomes the quencher.

Entropic changes control the charge separation process in triads mimicking photosynthetic charge separation.

Laser-induced optoacoustic spectroscopy (LIOAS) measurements with carotene-porphyrin-acceptor "supermolecular" triads (C-P-A, with A = C60, a naphthoquinone NQ, and a naphthoquinone derivative, Q) were carried out with the purpose of analyzing the thermodynamic parameters for the formation and decay of the respective long-lived charge separated state C*+-P-A*-. The novel procedure of inclusion of the benzonitrile solutions of the triads in Triton X-100 micelle nanoreactors suspended in water permitted the separation of the enthalpic and structural volume change contributions to the LIOAS signals, by performing the measurements in the range 4-20 degrees C. Contractions of 4.2, 5.7, and 4.2 mL mol-1 are concomitant with the formation of C*+-P-A*- for A = C60, Q and NQ, respectively. These contractions are mostly attributed to solvent movements and possible conformational changes upon photoinduced electron transfer, due to the attraction of the oppositely charged ends, as a consequence of the giant dipole moment developed in these compounds upon charge separation ( approximately 110 D). The estimations combining the calculated free energies and the LIOAS-derived enthalpy changes indicate that entropy changes, attributed to solvent movements, control the process of electron transfer for the three triads, especially for C-P-C60 and C-P-Q. The heat released during the decay of 1 mol of charge separated state (CS) is much smaller than the respective enthalpy content obtained from the LIOAS measurements for the CS formation. This is attributed to the production of long-lived energy storing species upon CS decay.

A large entropic term due to water rearrangement is concomitant with the photoproduction of anionic free-base porphyrin triplet states in aqueous solutions.

The enthalpy change, DeltaTH, and volume change, DeltaTV, associated with triplet state formation upon excitation of free-base meso-tetra-(4-sulfonatophenyl)porphyrin, TSPP4-, its Zn derivative, ZnTSPP4-, and meso-tetra-(4-carboxyphenyl)porphyrin, TCPP4-, were obtained in aqueous solutions by the application of laser-induced optoacoustics spectroscopy in the presence of phosphate salts of various monovalent cations (Li+, Na+, K+, NH4+ and Cs+). A linear correlation was found between DeltaTH and DeltaTV at different phosphate concentrations for the free-base porphyrins. The intercepts (132 +/- 8 kJ mol(-1) for TSPP4- and 164 +/- 23 kJ mol(-1) for TCPP4-) of these plots correspond to the respective value of the triplet energy content obtained from phosphorescence at 77 K (140 and 149 kJ mol(-1)). This suggests that DeltaTG for the triplet state formation is independent of the medium and an enthalpy-entropy compensation is responsible for the much smaller and salt-dependent DeltaTH values obtained at room temperature. The Gibbs energy for triplet state formation of the free-base porphyrins at room temperature is thus mainly determined by the entropic term due to solvent rearrangement. The DeltaTH values for 3ZnTSPP4- at different buffer concentrations and different cations are all between 130 and 150 kJ mol(-1), close to the triplet energy obtained from phosphorescence (E(T) = 155 kJ mol(-1)). The solvent structure and the nature of the counterion have a negligible influence on the 3ZnTSPP4-formation due to the blockage of the electron pairs on the central N atoms. Thus, the small DeltaTV value should be due to intrinsic bond changes upon 3ZnTSPP4- formation and no correlation between DeltaTH and DeltaTV should be expected in this case. The enthalpy change determines the Gibbs energy for 3ZnTSPP4-formation at room temperature.

Functional and biochemical analysis of the N-terminal domain of phytochrome A.

Phytochrome A (phyA) is a versatile plant photoreceptor that mediates responses to brief light exposures (very low fluence responses, VLFR) as well as to prolonged irradiation (high irradiance responses, HIR). We identified the phyA-303 mutant allele of Arabidopsis thaliana bearing an R384K substitution in the GAF subdomain of the N-terminal half of phyA. phyA-303 showed reduced phyA spectral activity, almost normal VLFR, and severely impaired HIR. Recombinant N-terminal half oat of PHYA bearing the phyA-303 mutation showed poor incorporation of chromophore in vitro, despite the predicted relatively long distance (>13 A) between the mutation and the closest ring of the chromophore. Fusion proteins bearing the N-terminal domain of oat phyA, beta-glucuronidase, green fluorescent protein, and a nuclear localization signal showed physiological activity in darkness and mediated VLFR but not HIR. At equal protein levels, the phyA-303 mutation caused slightly less activity than the fusions containing the wild-type sequence. Taken together, these studies highlight the role of the N-terminal domain of phyA in signaling and of distant residues of the GAF subdomain in the regulation of phytochrome bilin-lyase activity.