The use of spectroscopic techniques has shown that human serum albumin (HSA) undergoes reversible self-aggregation through protein–protein interactions. It ensures the subsequent overlapping of electron clouds along with the stiffening of the conformation of the interpenetrating network of amino acids of adjacent HSA molecules. The HSA oxidation process related to the transfer of one electron was investigated by pulse radiolysis and photochemical methods. It has been shown that the irradiation of HSA solutions under oxidative stress conditions results in the formation of stable protein aggregates. The HSA aggregates induced by ionizing radiation are characterized by specific fluorescence compared to the emission of non-irradiated solutions. We assume that HSA dimers are mainly responsible for the new emission. Dityrosine produced by the intermolecular recombination of protein tyrosine radicals as a result of radiolysis of an aqueous solution of the protein is the main cause of HSA aggregation by cross-linking. Analysis of the oxidation process of HSA confirmed that the reaction of mild oxidants (${\mathrm{Br}}_2^{•-}$, ${\mathrm{N}}_3^{•}$, ${\mathrm{SO}}_4^{•-}$) with albumin leads to the formation of covalent bonds between tyrosine residues. In the case of ^•OH radicals and partly, ${\mathrm{Cl}}_2^{•-}$, species other than DT are formed. The light emission of this species is similar to the emission of self-associated HSA.

We have shown that many proteins and enzymes (ovalbumin, β-lactoglobulin, lysozyme, insulin, histone, papain) undergo concentration-dependent reversible aggregation as a result of the interaction of the studied biomolecules. Moreover, irradiation of those protein or enzyme solutions under oxidative stress conditions results in the formation of stable soluble protein aggregates. We assume that protein dimers are mainly formed. A pulse radiolysis study has been made to investigate the early stages of protein oxidation by ${\mathrm{N}}_3^{•}$ or ^•OH radicals. Reactions of the ${\mathrm{N}}_3^{•}$ radical with the studied proteins lead to the generation of aggregates stabilized by covalent bonds between tyrosine residues. The high reactivity of the ^•OH with amino acids contained within proteins is responsible for the formation of various covalent bonds (including C–C or C–O–C) between adjacent protein molecules. In the analysis of the formation of protein aggregates, intramolecular electron transfer from the tyrosine moiety to Trp^• radical should be taken into account. Steady-state spectroscopic measurements with a detection of emission and absorbance, together with measurements of the dynamic scattering of laser light, made it possible to characterize the obtained aggregates. The identification of protein nanostructures generated by ionizing radiation using spectroscopic methods is difficult due to the spontaneous formation of protein aggregates before irradiation. The commonly used fluorescence detection of dityrosyl cross-linking (DT) as a marker of protein modification under the influence of ionizing radiation requires modification in the case of the tested objects. A precise photochemical lifetime measurement of the excited states of radiation-generated aggregates is useful in characterizing their structure. Resonance light scattering (RLS) has proven to be an extremely sensitive and useful technique to detect protein aggregates.

The reactions of radicals with human serum albumin (HSA) under reductive stress conditions were studied using pulse radiolysis and photochemical methods. It was proved that irradiation of HSA solutions under reductive stress conditions results in the formation of stable protein aggregates. HSA aggregates induced by ionizing radiation are characterized by unique emission, different from the UV emission of non-irradiated solutions. The comparison of transient absorption spectra and the reactivity of hydrated electrons ($e_{aq}^{-}$) with amino acids or HSA suggests that electron attachment to disulfide bonds is responsible for the transient spectrum recorded in the case of albumin solutions. The reactions of $e_{aq}^{-}$ and ${CO}_2^{•-}$ with HSA lead to the formation of the same products. Recombination of sulfur-centered radicals plays a crucial role in the generation of HSA nanoparticles, which are stabilized by intermolecular disulfide bonds. The process of creating disulfide bridges under the influence of ionizing radiation is a promising method for the synthesis of biocompatible protein nanostructures for medical applications. Our Raman spectroscopy studies indicate strong modification of disulfide bonds and confirm the aggregation of albumins as well. Low-temperature measurements indicate the possibility of electron tunneling through the HSA protein structure to specific CyS-SCy bridges. The current study showed that the efficiency of HSA aggregation depends on two main factors: dose rate (number of pulses per unit time in the case of pulse radiolysis) and the temperature of the irradiated solution.

This research investigated the influence of the layered structure of paper on its selected strength properties. Two types of cellulose pulp were used: unbleached kraft pine pulp (UBSK) and unrefined bleached birch kraft pulp (BHK). Multi-ply papers were obtained from layers of appropriate grammage formed separately and bonded in a wet state. Paper layers were formed from primary pulps (UBSK and BHK) or their mixtures (three-ply UBSK-BHK paper and three-ply UBSK-BHK-UBSK paper), from which various variants of laboratory paper samples were made. This work focused on three-layer papers. The obtained laboratory sheets were analysed for changes in tear resistance, tensile strength, bursting strength, short span compression strength (SCT), and corrugating medium test (CMT). In most cases, three-layer structures were characterised by higher strength parameters than single-layer papers. The tear strength and SCT of papers consisting of three layers formed from both mixed pulps were slightly lower than those of solid papers. The results demonstrate the feasibility of producing three-ply paper with significantly reduced variability in parameters such as elongation and bursting strength, which are inherently subject to high variability.