Photosynthetic reaction center variants made via genetic code expansion show Tyr at M210 tunes the initial electron transfer mechanism

Photosynthetic reaction centers (RCs) from Rhodobacter sphaeroides were engineered to vary the electronic properties of a key tyrosine (M210) close to an essential electron transfer component via its replacement with site-specific, genetically encoded noncanonical amino acid tyrosine analogs. High fidelity of noncanonical amino acid incorporation was verified with mass spectrometry and X-ray crystallography and demonstrated that RC variants exhibit no significant structural alterations relative to wild type (WT). Ultrafast transient absorption spectroscopy indicates the excited primary electron donor, P*, decays via a ∼4-ps and a ∼20-ps population to produce the charge-separated state P+HA− in all variants. Global analysis indicates that in the ∼4-ps population, P+HA− forms through a two-step process, P*→ P+BA−→ P+HA−, while in the ∼20-ps population, it forms via a one-step P* → P+HA− superexchange mechanism. The percentage of the P* population that decays via the superexchange route varies from ∼25 to ∼45% among variants, while in WT, this percentage is ∼15%. Increases in the P* population that decays via superexchange correlate with increases in the free energy of the P+BA− intermediate caused by a given M210 tyrosine analog. This was experimentally estimated through resonance Stark spectroscopy, redox titrations, and near-infrared absorption measurements. As the most energetically perturbative variant, 3-nitrotyrosine at M210 creates an ∼110-meV increase in the free energy of P+BA− along with a dramatic diminution of the 1,030-nm transient absorption band indicative of P+BA– formation. Collectively, this work indicates the tyrosine at M210 tunes the mechanism of primary electron transfer in the RC.

High Resolution Powder Electron Diffraction in Scanning Electron Microscopy

A modern scanning electron microscope equipped with a pixelated detector of transmitted electrons can record a four-dimensional (4D) dataset containing a two-dimensional (2D) array of 2D nanobeam electron diffraction patterns; this is known as a four-dimensional scanning transmission electron microscopy (4D-STEM). In this work, we introduce a new version of our method called 4D-STEM/PNBD (powder nanobeam diffraction), which yields high-resolution powder diffractograms, whose quality is fully comparable to standard TEM/SAED (selected-area electron diffraction) patterns. Our method converts a complex 4D-STEM dataset measured on a nanocrystalline material to a single 2D powder electron diffractogram, which is easy to process with standard software. The original version of 4D-STEM/PNBD method, which suffered from low resolution, was improved in three important areas: (i) an optimized data collection protocol enables the experimental determination of the point spread function (PSF) of the primary electron beam, (ii) an improved data processing combines an entropy-based filtering of the whole dataset with a PSF-deconvolution of the individual 2D diffractograms and (iii) completely re-written software automates all calculations and requires just a minimal user input. The new method was applied to Au, TbF3 and TiO2 nanocrystals and the resolution of the 4D-STEM/PNBD diffractograms was even slightly better than that of TEM/SAED.

SECONDARY ELECTRON EMISSION FROM THIN ALUMINIUM FOILS PRODUCED BY HIGH ENERGY ELECTRON BEAMS

The description of the experimental equipment and technique for measuring the secondary emission of elec-trons (SEE) with application of accelerated electrons at the linear accelerator of the IHEPNP NSC KIPT with ener-gies up to 30 MeV and a standard secondary emission monitor [1] are presented. Experimental data of secondary electron emission yields from thin aluminum targets (8 and 50 μm) for primary electron beam energies of 16 and 25 MeV have been experimentally measured. The analysis of the experimental data and their comparison with the theory are carried out. It is shown that the proposed technique for measuring the yields of secondary electron emis-sion is useful and applied for study of low-energy and δ-electrons yields from thin foils, as well as to research the effect of the density effect depending on the energy of the primary electron beam.

FORMATION AND MONITORING OF SECONDARY X-RAY RADIATION UNDER PRODUCT PROCESSING WITH ELECTRON BEAM

When conducting an industrial radiation processes at an electron accelerator, a part of the beam energy is trans-formed into bremsstrahlung radiation. In such a way, the mixed e,X-radiation is formed in the area behind an irra-diated object. The intensity of the electron and photon components in the radiation is determined by the energy and power of the primary electron beam, as well as by the parameters of the object and devices located behind it. In paper, the characteristics of the e,X-radiation accompanying the product processing by a scanning electron beam with energy 8…12 MeV at a LU-10 Linac of NSC KIPT are studied. The conditions for obtaining a source of sec-ondary X-rays in the state of electronic equilibrium, as well as its monitoring using an extended free-air ionization chamber are explored. Such an extra-source of radiation can be used for carrying out various non-commercial pro-grams like radiation tests, sanitization of archival materials and cultural heritage objects, etc.

Application of Prandtl’s Theory in the Design of an Experimental Chamber for Static Pressure Measurements

Pumping in vacuum chambers is part of the field of environmental electron microscopy. These chambers are separated from each other by a small-diameter aperture that creates a critical flow in the supersonic flow regime. The distribution of pressure and shock waves in the path of the primary electron beam passing through the differentially pumped chamber has a large influence on the quality of the resulting microscope image. As part of this research, an experimental chamber was constructed to map supersonic flow at low pressures. The shape of this chamber was designed using mathematical–physical analyses, which served not only as a basis for the design of its geometry, but especially for the correct choice of absolute and differential pressure sensors with respect to the cryogenic temperature generated in the supersonic flow. The mathematical and physical analyses presented here map the nature of the supersonic flow with large gradients of state variables at low pressures at the continuum mechanics boundary near the region of free molecule motion in which the Environmental Electron Microscope and its differentially pumped chamber operate, which has a significant impact on the resulting sharpness of the final image obtained by the microscope. The results of this work map the flow in and behind the Laval nozzle in the experimental chamber and are the initial basis that enabled the optimization of the design of the chamber based on Prandtl’s theory for the possibility of fitting it with pressure probes in such a way that they can map the flow in and behind the Laval nozzle.

Direct Observation of Ultrafast Exciton Localization in an Organic Semiconductor with Soft X-ray Transient Absorption Spectroscopy

We report the first demonstration of time-resolved X-ray absorption spectroscopy to track previously undetected photoinduced dynamics of a paradigmatic crystalline conjugated polymer: poly(3-hexylthiophene) (P3HT) commonly used in solar cell devices. The pi to pi* transition, the first step of solar energy conversion, is pumped with a 15 fs optical pulse and the dynamics are probed by an attosecond soft X-ray pulse at the carbon K-edge. We observe direct spectroscopic signatures of the initially hot excitonic state, which is delocalized over multiple polymer chains, undergoing a rapid evolution on a sub 50 fs timescale which can be directly associated with cooling and localization to form the lowest excitonic state on a single polymer chain. This sensitivity of time-resolved X-ray spectroscopy to the primary electron dynamics occurring directly after excitation paves the way for new insights in a wide range of organic optoelectronic materials.

Shedding Light on Primary Donors in Photosynthetic Reaction Centers

Chlorophylls (Chl)s exist in a variety of flavors and are ubiquitous in both the energy and electron transfer processes of photosynthesis. The functions they perform often occur on the ultrafast (fs–ns) time scale and until recently, these have been difficult to measure in real time. Further, the complexity of the binding pockets and the resulting protein-matrix effects that alter the respective electronic properties have rendered theoretical modeling of these states difficult. Recent advances in experimental methodology, computational modeling, and emergence of new reaction center (RC) structures have renewed interest in these processes and allowed researchers to elucidate previously ambiguous functions of Chls and related pheophytins. This is complemented by a wealth of experimental data obtained from decades of prior research. Studying the electronic properties of Chl molecules has advanced our understanding of both the nature of the primary charge separation and subsequent electron transfer processes of RCs. In this review, we examine the structures of primary electron donors in Type I and Type II RCs in relation to the vast body of spectroscopic research that has been performed on them to date. Further, we present density functional theory calculations on each oxidized primary donor to study both their electronic properties and our ability to model experimental spectroscopic data. This allows us to directly compare the electronic properties of hetero- and homodimeric RCs.

Electron and Photon Energy Spectra Outside of 6 MV X-ray Small Radiotherapy Field Edges Produced by a Varian iX Linac

Due to the increase in the survival probability for patients treated with modern radiotherapy techniques to live enough for experimenting the late radiation effect, low dose outside the treatment volume becomes a concern. However, besides the absorbed dose, the beam quality outside the field edge should be taken into account. This work aimed at investigating the photon and electron fluence spectra outside the field edges for several small radiotherapy fields for determining the quality of the beams in order to better evaluate the secondary effect after modern radiotherapy treatments. Phase-space files of a 6 MV X-ray beam produced by a Varian iX linac for eight small fields of 0.7 × 0.7 cm2, 0.9 × 0.9 cm2, 1.8 × 1.8 cm2, 2.2 × 2.2 cm2, 2.7 × 2.7 cm2, 3.1 × 3.1 cm2, 3.6 × 3.6 cm2, and 4.5 × 4.5 cm2 and for the reference 10 × 10 cm2 field at SSD = 100 cm were generated using the BEAMnrc code. The photon and electron fluences in each field were calculated at 0.15, 1.35, and 9.85 cm water depth and several off-axis distances using FLURZnrc. The number of low-energy electrons between 1 and 10 keV at 2 cm outside the field edge increases by 60% compared to the central axis. Due to the relatively high linear energy transfer (LET) of these electrons, the results of this work should help to better evaluate the possible late effect of secondary radiation on healthy organs close to the tumor volume after radiotherapy treatment. We also observed high-energy electrons outside the field edge that are attributed to the leakage of the primary electron beam from the head of the linac. From a standpoint of radiological protection, these electrons should be taken into account when evaluating the dose delivered to the patient’s skin.

Down-Regulation of Photosynthetic Electron Transport and Decline in CO2 Assimilation under Low Frequencies of Pulsed Lights

The decline in CO2 assimilation in leaves exposed to decreasing frequencies of pulsed light is well characterized, in contrast to the regulation of photosynthetic electron transport under these conditions. Thus, we exposed sunflower leaves to pulsed lights of different frequencies but with the same duty ratio (25%) and averaged light intensity (575 μmoles photons m−2 s−1). The rates of net photosynthesis Pn were constant from 125 to 10 Hz, and declined by 70% from 10 to 0.1 Hz. This decline coincided with (1) a marked increase in nonphotochemical quenching (NPQ), and (2) the completion after 25 ms of illumination of the first phase of P700 photooxidation, the primary electron donor of PSI. Under longer light pulses (<5 Hz), there was a slower and larger P700 photooxidation phase that could be attributed to the larger NPQ and to a resistance of electron flow on the PSI donor side indicated by 44% slower kinetics of a P700+ dark reduction. In addition, at low frequencies, the decrease in quantum yield of photochemistry was 2.3-times larger for PSII than for PSI. Globally, our results indicate that the decline in CO2 assimilation at 10 Hz and lower frequencies coincide with the formation of NPQ and a restriction of electron flows toward PSI, favoring the accumulation of harmless P700+.