Herewith, lymphocytes featured an increase in the activity of lactate dehydrogenase, a cytosolic enzyme of glycolysis, without changes in succinate-dehydrogenase activity. Structural changes occurring in the SN and striatum manifested themselves in the destruction of mitochondria, degeneration of neurons and synapses, and stratification of myelin sheaths in them. Subcutaneous injections of 30 µg/kg uridine for 22 days restored the neurotoxin-induced changes in these parameters to levels close to the control. 5-Hydroxydecanoate (5 mg/kg), a specific mitoK-ATP channel inhibitor, eliminated the beneficial effect of uridine for almost all characteristics tested, indicating the involvement of the mitoK-ATP channel in the protective effect of uridine. The mechanism of the protective effect of uridine and its therapeutic applications for the prevention and treatment of PD are discussed.Using a model of Parkinson’s disease (PD) induced by the bilateral injection of neurotoxin 6-hydroxydopamine (6-OHDA) into rat brain substantia nigra (SN), we showed uridine to exert a protective effect associated with activation of the mitochondrial ATP-dependent potassium (mitoK-ATP) channel. Injection of 4 µg neurotoxin evoked a 70% decrease in the time the experimental animal spent on the rod in the RotaRod test, an increase in the amount of lipid peroxides in blood serum and cerebral-cortex mitochondria and the rate of reactive oxygen species formation, and a decrease in Ca Parkinson’s disease (PD) is a neurodegenerative disorder the probability of the development of which significantly increases with age. It is expected that, as a result of overall aging of the world population, the number of patients with PD will significantly increase . ElucidaOne of the main processes underlying neurodegenerative disorders, in particular, PD, is mitochondrial dysfunction. Taking into account the high energy needs of the body, especially of the brain, this condition may have devastating consequences for the survival of neurons .The mitochondrial dysfunction in brain tissues is commonly caused by an excessive production of reactive oxygen species (ROS), whose level increases with age . At the Since PD patients are found to feature an inhibition of respiratory chain complex I in mitochondria , one of We have previously found that the injection of uridine to animals leads to an increase in the concentration of uridine diphosphate (UDP), a natural activator of the mitochondrial ATP-dependent potassium (mitoK-ATP) channel, which normalizes oxidative exchange and eliminates mitochondrial dysfunction ,9,10,11.In this work, we studied the therapeutic action of uridine in a model of neurotoxin-induced PD in rats and the degree of the involvement of the mitoK-ATP channel in the development of Parkinson’s syndrome.In a model of PD created by the bilateral injection of 6-OHDA into the rat brain SN, we observed a disturbance of the dopaminergic nigrostriatal pathway and an impairment of the motor function. Nissl staining used to quantitatively assess the fate of dopaminergic neurons in the substantia nigra pars compacta after administration of the neurotoxin revealed that 70% of the neurons died. Upon creation of the model, the rats were in a calm environment with free access to water and food; in the postoperative period, they were under daily control. Additionally, the animals were administered Ringer’s solution (5 mL per day). Motor impairments were assessed using the RotaRod behavioral test on the 22nd day after the injection of the neurotoxin A.Administration of the neurotoxin decreased by 70% the time spent by the animal on the rotating rod compared to the control group A. To corDisturbance of calcium homeostasis observed in PD is known to lead to cell death. The cell death is, presumably, associated with the overload of mitochondria with calcium, which promotes the development of oxidative stress in them and changes the permeability of the mitochondrial membrane followed by the opening of a nonspecific pore and the death of dopaminergic neurons ,13.The results of the present study showed that the administration of the neurotoxin to model PD decreased the calcium retention capacity of mitochondria by 63% relative to the control. Injection of uridine at a concentration of 30 mg/kg of body weight together with 6-OHDA increased the calcium retention capacity of mitochondria to the control level. Application of 5-HD completely eliminated the beneficial action of uridine B.The rate of hydrogen peroxide formation in the experiments was determined in a system with Amplex Red and horseradish peroxidase. It was found that the neurotoxin increased the rate of hydrogen peroxide formation in rat brain mitochondria in the experimental group by 23% compared to the control groups. The injection of uridine together with the neurotoxin decreased the characteristic to values even below the control, and the injection of 5-HD in combination with uridine eliminated the protective effect of uridine, increasing the rate of hydrogen peroxide formation A.When diffusing through biological membranes, hydrogen peroxide can react with unsaturated fatty acids of lipids, which leads to the destruction of membranes. One of the products of these reactions is malondialdehyde (MDA). In this work, we determined the content of lipid peroxidation products in brain mitochondria B. In ratThe measurements showed that the MDA content in the blood serum of rats with PD increased by 37%, and the treatment with uridine lowered this characteristic to the control level C. At theA cytobiochemical determination of the activity of the key mitochondrial enzyme, succinate dehydrogenase (SDH), in immobilized blood lymphocytes in a blood smear from PD rats did not reveal any significant differences from the control. After the injection of uridine to sick animals, the activity of SDH increased by 80% compared to the control. The application of 5-HD reduced the beneficial effect of uridine. Measurements of the activity of the glycolysis enzyme, lactate dehydrogenase (LDH), in blood lymphocytes in a smear from PD rats showed that it increased by 35% compared to the control, and the treatment with uridine decreased the activation to that of the control value. The administration of the 5-HD inhibitor abolished the effect of uridine, increasing the LDH level by 37% to the level in the group of sick rats. Thus, in the mitochondria of blood lymphocytes from rats with PD, glycolysis dominated over oxidative phosphorylation; the administration of uridine enhanced mitochondrial respiration by 80% and reduced glycolysis. The injection of 5-HD completely abolished the effect of uridine .r = 0.99. In animals with Parkinson’s syndrome, the LDH activity increased by 37%, and the MDA content rose by 35%. Treatment with uridine decreased both characteristics to the values for the control group, and 5-HD completely abolished the effect of uridine in lymphocytes and serum. The reduction in SDH activity and the increase in LDH indicate the disturbance of energetic processes in lymphocytes in PD, which shows up in the retardation of aerobic oxidation and enhancement of glycolysis; this is in line with the literature data [The characteristics of glycolysis (activity of LDH) in lymphocytes and oxidative stress (MDA level) in serum were revealed to have a direct positive correlation with coefficient ure data .Images of the ultrastructure of the dopaminergic neuron in the SN were obtained 22 days after the induction of Parkinson’s syndrome in rats by the bilateral injection of 6-OHDA into the SN .Exposure to the neurotoxin resulted in the appearance of a large number of dark neurons B. As comIn the neuropil of SN neurons, we observed significant structural damage to the myelin sheaths of axons B,D.Compared with the control A, in neuAlong with disruptions in the SN structure, we observed changes in the striatum. They were in many respects similar to those found in the SN and consisted in the appearance of dark neurons with a dark cytoplasm and a dark nucleus, which had an untwisted sheath .The cisternae of the reticulum and the Golgi complex in the cytoplasm were dilated B as compThe termini of axons of dopaminergic neurons of the SN commonly form axo-dendritic synapses in the striatum. Analysis of the state of axons in the striatum in control A and expIn experimental rats, dilated axo-dendritic synaptic endings with significant accumulation of synaptic vesicles were found C. SynaptThe structure of myelin sheaths in the neuropil of striatal neurons was also disordered in the rat PD model .Sheaths with stratified lamellae and membrane protrusions appeared B. After The work proposes a new approach to the prevention and treatment of PD. The approach is based on the activation of the mitoK-ATP channel by its natural activator, which leads to an enhancement of energy metabolism, normalization of oxidative metabolism in brain tissues, and, as a consequence, a decrease in mitochondrial dysfunction developing in this disease.A characteristic sign of PD is the progressive degeneration of dopaminergic neurons in the compact part of the SN and striatum, which leads to a decline in the motor activity of animals ,7. UridiPD is also characterized by an oxidative metabolism disturbance in brain tissues expressed in the accumulation of ROS, as well as by an increase in the rate of hydrogen peroxide production. This is probably due to the fact that, being structurally similar to dopamine, 6-OHDA enters the cell by means of dopamine and norepinephrine transporters, where it increases the concentration of ROS both enzymatically  and nonenzymatically . FurtherWe used uridine to correct biochemical impairments caused by the administration of 6-OHDA. At the concentrations applied, this nucleoside prevents the development of hypoxia in tissues, as shown in our previous research . AdminisModeling of PD in rats revealed significant changes in the structures of rat brain SN and striatum. Each of these regions is of critical significance for the development of PD. Their structures are part of the extrapyramidal motor system, which is responsible for the regulation of motor function and muscle tone and is involved in many vegetative functions, such as respiration, cardiac activity, and blood vessel tone .Morphological changes are observed in neurons, presynaptic endings, and in the myelin sheath of the SN and striatum. These changes may be a cause of a decrease in energy efficiency and ATP level in brain tissues, which leads to progressive degeneration of dopaminergic neurons in the compact region of the SN and striatum . An incrIt is considered that neurodegeneration begins in synaptic junctions, as they are the first to be damaged in PD . An incrUpon administration of the neurotoxin, we observed a disruption of myelin sheaths known to be required for the separation of nerves from one another and the enhancement of nerve pulse transmission. At these structural disorders, signals become mixed, and the normal movements of the animal become impossible . The abom) [m diminishes the rate of peroxide formation in mitochondria sevenfold [The study using the inhibitory analysis shows that the mechanism by which uridine affects the biochemical parameters and structure of experimental animals’ brains is associated to a greater extent with the activation of the mitoK-ATP channel, because 5-HD, the channel inhibitor, abolishes the positive effect of uridine almost completely. We explain this by an increase in the concentration of UDP, an activator of this channel, in tissues at the administration of uridine . It is km) ,26. It hevenfold .As the effect of uridine on brain tissues is almost completely eliminated by 5-HD, a selective inhibitor of the mitoK-ATP channel, we suggest that this channel is involved in maintaining metabolic processes and integrity of the brain structure in PD and, probably, in the development of the disease. The assumption about the role of the channel in the development of PD is consistent with our earlier data on a significant (3–5-fold) decrease in the activity of this channel with age . The fac2, known to be depleted during the development of oxidative stress in toxic models of PD [Still, as 5-HD did not always completely abolish the effect of uridine, other metabolic pathways of this nucleoside can be also assumed to be involved in protecting the rat body from the neurotoxin. One of such pathways, as we noted above, can be the synthesis of acetyl-CoA from uridine, which is important for the recovery of the Krebs cycle energy equivalents, NADH and FADNls of PD . In thisls of PD ,31. As fls of PD ,9. An inls of PD .Since the mitoK-ATP channel inhibitor eliminated the beneficial effect of uridine in almost all cases, we assume that the main mechanism of action of this metabolite is through the activation of the channel. The activation of the channel leads to an acceleration of potassium cyclization in mitochondria, which lowers their membrane potential to induce a “mild uncoupling”. Herewith, the rate of peroxide formation in mitochondria decreases, oxidative metabolism normalizes, and both the calcium overload of cells and the neurotoxin-induced structural changes in the brain tissue are prevented.Wistar male rats (N = 23) weighing 190–220 g were used. The animals were housed in a vivarium at the Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences. All manipulations with animals were carried out in compliance with the Council Directive 2010/63 EU of the European Parliament and the Regulations of the European Convention for the Protection of Vertebrate Animals Used for Scientific Purposes .In this series of experiments, animals were divided into four groups: (1) the control group; to the SN of both brain hemispheres, 4 µL of a 0.2% L-ascorbic acid solution was injected (N = 5); (2) the 6-OHDA group; 4 µg of neurotoxin 6-OHDA dissolved in 6 µL of a 0.2% L-ascorbic acid solution was injected (N = 6); (3) the 6-OHDA + U group; after the injection of 6-OHDA, the mitoK-ATP channel activator, uridine (U), was daily administered intraperitoneally for 22 days at a dose of 30 mg/kg of body weight (N = 6); (4) the 6-OHDA+U + 5-HD group; after the injection of 6-OHDA, uridine (U) was administered daily, and 15 min after the injection of uridine, the mitoK-ATP channel inhibitor, 5-HD, was subcutaneously injected at a dose of 5 mg/kg of body weight (N = 6).For anesthesia, Xylozin (4 mg/kg of body weight) and Zoletil (60 mg/kg) were used. 6-OHDA (5 mg) was dissolved in 0.8 mL of 300 mM sodium chloride  with 0.1% ascorbic acid. Operations were carried out on a stereotaxic frame using a 10 mL micro syringe  by the method described by Torres and Dunnett . The neuThe endurance and motor coordination of animals were assessed on a RotaRod device  on the 22nd day after surgery. The facility was a rotating horizontal rod with lateral vertical baffles. An experimenter placed a rat into the facility and controlled the running of the animal. The time during which the animal stayed on the rotating rod (speed 8 cm/s) was measured.g for 10 min. The supernatant was poured into a new test tube and centrifuged again at 12,000× g for 10 min. Then, the supernatant was removed and the sediment was homogenized to a homogenous state in a washing medium  and centrifuged at 12,000× g for 10 min. The resulting sediment was collected and further homogenized with the washing medium. All procedures were carried out on ice in a cold room at +4 °C.Rats were decapitated on the 22nd day after surgery. Brain tissue fragments were washed and homogenized by a manual homogenizer in an isolation medium . Homogenates were centrifuged at 12,000× 2+ retention capacity of mitochondria from the brain of rats in the norm and in experimental parkinsonism was determined using a Record-4 potentiometric system with a Ca2+-selective electrode . The incubation medium contained 150 mM KCl, 1 mM NaH2PO4, 10 µM EGTA, and 10 mM HEPES-KOH (pH 7.4). The concentration of the mitochondrial protein in a cuvette was 1 mg/mL. The substrates used were 5 mM malate and 5 mM potassium gluconate. CaCl2 was added in fractions of 25 µM every 30 s. The calcium retention capacity was calculated as nmol Ca2+/mL × mg protein.The Ca2PO4, 10 mM HEPES-KOH, and 0.5 mM EGTA, pH 7.4. The hydrogen peroxide production was measured at 37 °C in the presence of 20 µM Amplex Red and 2 U/mL horseradish peroxidase. To avoid the impact of optical effects associated with mitochondrial swelling, the activity of mitochondria was measured using the two-wave mode of registration (572–600 nm) [2O2/min × mg protein.The generation of hydrogen peroxide by rat brain mitochondria was measured using a fluorimeter  at an excitation of 570 nm and an emission of 585 nm. The reaction mixture contained 120 mM KCl, 5 mM KH–600 nm) . The con3COONa, 300 µL of a 0.8% solution of thiobarbituric acid (TBA), and 50–25 µL serum or mitochondria. Eppendorf dishes with the reaction mixture were incubated for 1 h at 90 °C. Then, the liquid was centrifuged at 2000× g for 5 min. The product was a trimethine complex of characteristic pink color (λmax = 532 nm). The measurement was performed on a spectrophotometer  at 532–650 nm. The results were expressed in nmol/min × mg protein.The concentration of lipid peroxidation products in rat brain mitochondria and blood serum was determined from the reaction of 250–275 µL of 5% SDS, 900 µL of 20% CH2) in a sample of 100 lymphocytes.The activities of succinate dehydrogenase (SDH) in mitochondria in peripheral blood and lactate dehydrogenase (LDH) in the cytosol in lymphocytes in peripheral blood were determined by a biocytochemical method  from the reduction in nitro blue tetrazolium (NBT) to dark blue formazan [After decapitation, pieces of the SN were taken from rats. The material was fixed immediately, by placing tissue samples in a 2.5% solution of glutaraldehyde in 0.1 M cacodylate buffer (pH 7.4). Postfixation was performed with 2% osmium tetroxide in 0.1 M cacodylate buffer (pH 7.4). Dehydration was carried out in alcohols of increasing concentration and acetone. Dehydrated samples were embedded in Epon 812 resin. Ultrathin sections 60–70 nm thick were made on an ultramicrotome . Sections were contrasted with 1% uranyl acetate and zinc citrate. Samples were scanned using a transmission electron microscope . The total number of mitochondria (nMch) on microphotographs was calculated.t-test. Differences are significant at * p < 0.05 and ** p < 0.01. MS Excel, ImageJ , Origin 2016 , and Prism GraphPad 7  softwares were used for data and statistical analysis.All values are given as the means ± standard error of the means. Statistical analysis was performed by the paired Based on the data obtained, it was concluded that the natural metabolite uridine protects the rat body against impairments induced by the administration of neurotoxin 6-OHDA and prevents negative consequences observed when modeling PD. The mechanism of the protective effect of uridine is mainly associated with the activation of the mitoK-ATP channel, which reduces the oxidative stress and the mitochondrial dysfunction in tissues. In this connection, uridine can be considered to be a metabolite that prevents the development of PD and can be used as a therapeutic means at its early stages.


Particle media are widely used in engineering and greatly influence the performance of engineering materials. Asphalt mixtures are multi-phase composite materials, of which coarse aggregates account for more than 60%. These coarse aggregates form a stable structure to transfer and disperse traffic loads. Therefore, knowing how to adjust the structural composition of coarse aggregates to optimize their performance is the key to optimize the performance of asphalt mixtures. In this study, the effects of different roughness and different sizes on the interlocking force and contact force of coarse aggregates were investigated through means of simulation (DEM), and then the formation-evolution mechanism of the coarse aggregate structure and the role of different sizes of aggregates in the coarse aggregate structure were analyzed. Subsequently, the optimal ratio of coarse aggregates was explored through indoor tests, and finally, the gradation of asphalt mixture based on the optimization of fine structure was formed and verified through indoor tests. The results showed that the major model can effectively reveal the role of different types of aggregates in the fine structure and the relationship between the strength of contact forces between them and clarify that the strength of the fine structure increases with the increase in aggregate roughness. Hence, the coarse aggregate structure can be regarded as a contact force transmission system composed of some strong and sub-strong contact forces. Their formation-evolution mechanism can be regarded as a process of the formation of strong and sub-strong contact forces and the transformation from sub-strong contact force to strong contact force. Moreover, the dynamic stability of the optimized graded asphalt mixture was increased by 30%, and the fracture toughness was increased by 26%. As the most widely used road pavement materials, the asphalt mixture plays an important role in infrastructure construction. In the asphalt mixture as an asphalt–void–aggregate composition of multi-phase complex materials , its coarse aggregates account for more than 60%, and these coarse aggregates can form a stable structure to transfer and disperse the traffic load. Therefore, there is a significant correlation between coarse aggregate and the performance of the asphalt mixture. Shen et al. ,2 specifShashidhar et al.  applied Currently, only a few studies have analyzed the structural mechanical properties of aggregates. Cal et al.  establisHence, this study analyzed the macro-skeleton interlocking forces and meso-aggregate contact force of the coarse aggregate structure based on the DEM simulation and clarified the formation–evolution mechanism and the function of different types of aggregates on the coarse aggregate structure. On this basis, the coarse aggregate structure is optimized based on the principle of optimal aggregate skeleton strength, and a better asphalt mixture gradation is proposed, which passed the indoor test verification.In this paper, AH-70 asphalt is adopted, and the basic performance of AH-70 asphalt is tested according to the specification . Its indThe physical and mechanical properties of the aggregate affect the road characteristics of the asphalt mixture. Coarse aggregate is mainly used to form the skeleton structure, fine aggregate is mainly used to fill the skeleton structure of coarse aggregate, mineral powder and asphalt to form asphalt slurry filled in the gaps between coarse and fine aggregate and the aggregate bonded together, so that the asphalt mixture to produce the ability to resist the traffic load. The aggregate is obtained from limestone in Shandong Province, China. The test results  of the bDue to the particularity of DEM numerical calculation, the complexity of model operation depends on the number of particles and contact points. The DEM simulation of unbonded particles is usually random, and these particles need to reach the equilibrium state of contact force under gravity before the calculation begins. However, the existing numerical models ,19,20 iniS) of coarse aggregate is calculated according to the mass, apparent density and size of different coarse aggregates, as shown in Equation (1). Then, the built-in command “ball” of PFC2D is used to generate circular particles in the simulated cylinder and make them meet the particle size requirements of a coarse aggregate.The two-dimensional mapping area  of the coarse aggregate, four sub-particles are generated by the built-in command “ball” of PFC2D. The structure of irregular particles is shown in Secondly, the built-in command “b_id” of PFCS) can be represented by Equation (2). Then, S is set qual to the initial circular particle area (oS), as shown in Equation (3). On this basis, the radius of the particle is obtained by Equation (4).As shown in Subsequently, the center coordinates of the four small particles are calculated according to Equations (5)–(8).a values are shown in a value.A series of particles of different 2D. Then, initial particles of number t are removed by using “delete” in the PFC2D and replaced by the corresponding irregular particles. When the aggregate structure reaches the static equilibrium state under the action of gravity, the numerical model of the irregular particle structure can be obtained, as shown in In the end, the irregular particles are composed of four sub-particles by using “clump” in the PFCIn addition, the simplified Hertz–Mindlin constitutive is adopted because it can better describe the nonlinear characteristics of aggregate structures, as shown in Equations (9)–(11).According to In order to simulate the confining pressure effect in the test, the speed of walls  are controlled by the servo mechanism. When the aggregate structure is compacted to reach the target porosity, the servo mechanism of the W1 and W4 stops running, while that of the W2 and W3 keeps running during the whole numerical test. The servo mechanism can be listed as follows:The simulated indenter is penetrated into the numerical model of aggregate structure at a constant speed, and the contact force and displacement of the indenter are recorded during the whole simulated penetration process. The numerical simulation results are shown in (1)The relationship between force and displacement of the indenter(2)Initial stageIt can be seen that the particle contact force is uniformly distributed in (3)Peak stageThe particle contact force is not uniformly distributed under the action of load and has an obvious phenomenon of concentration of contact force, as shown in (4)Attenuation stageSubsequently, when the contact force of the indenter exceeds the peak value, the aggregate structure begins to collapse, which corresponds to the curve state of B-C. Correspondingly, in Based on the conventional triaxial test system, this paper reproduces the above numerical simulation loading process by setting an improved loading head to measure the interlocking forces of the aggregate structure. The schematic diagram of the test device is shown in a value and follows the principle of the highest simulation accuracy. The comparison of simulation results and test results of aggregate structures with different a values is shown in The interlocking forces of the aggregate structure are evaluated by using a series of laboratory and numerical tests. As shown in Equations (9)–(11), three DEM contact parameters  need to be assigned to the numerical model of aggregate structure. The shear modulus and Poisson’s ratio can be measured by corresponding tests. However, the friction coefficient is obtained through the indoor test inversion after the change in a = 0.6) have the best accuracy, with an average error of only 4.9%. Hence, the parameters corresponding to a = 0.6  are used in this section for the later numerical simulations. In addition, the same parameters are used for other a value to facilitate the comparison between the simulation results.As shown in However, the above simulations are also flawed in that 2D aggregate structural bodies are referenced instead of 3D structural bodies, which are known to affect the contact force paths. Moreover, 3D model has the problems of long simulation time and high complexity. Many existing studies ,25,26 ofa values on the mechanical properties of the aggregate structures are discussed. There are five kinds of aggregates, including 13.2~19 mm (D1), 9.5~13.2 mm (D2), 4.75~9.5 (D3), 2.36~4.75 mm (D4) and 1.18~2.36 mm (D5). In addition, the a value also includes five kinds, which are 0, 0.2, 0.4, 0.6 and 0.8. The simulation plan is as follows:In this section, the effect of different aggregates and Step 1: Denote the numerical model of D1 mixing as P1 and record the corresponding aggregate interlocking force and contact force.Step 2: Obtain the aggregate interlocking force and contact force of the aggregate structure mixed with different proportions of P1 and D2; determine the optimal ratio of P1 and D2 by following the principle of maximum aggregate interlocking force and denote the above ratio of P1 and D2 as P2.Step 3: Denote the optimal ratio of P2 and D3 as P3 by following the above principle and record the corresponding force.Step 4: By analogy, complete the design of P4 and P5 and record the corresponding results.f) of P1~P5 and the particle size (mind) of the minimum-size aggregate in the aggregate structure is shown in f and a.The relationship between the aggregate interlocking force , slightly strong , strong , where {F} is the average value of the aggregate contact force.The aggregate contact forces play an important role in the strength of granular media. In this section, the distribution and number of aggregate contact forces in the numerical model of aggregate structure are studied when the aggregate structure reaches the ultimate load strength. The aggregate contact force is divided into three levels : weak (0a = 0.6). a values. The distribution probability of aggregate contact force [P(f)] is given by Equation (13).As shown in With the addition of D5, the number of weak contact forces in the aggregate structure increases sharply, while the number of strong and slightly strong contact forces increases very slowly. This leads to a decrease in the distribution probability of strong and slightly strong contact forces, while the distribution probability of weak contact force increases obviously. Furthermore, it can be seen from the above research that P5 and P4 have little difference in aggregate interlocking forces. This indicates that the strong and slightly strong contact forces are mainly provided by D1~D4. The weak contact force has little effect on the strength enhancement of aggregate structure. The D5 type mainly provides weak contact force for the aggregate structure but makes little contribution to strong and slightly strong contact forces. This explains why the interlocking force of the aggregate structure does not increase obviously with the addition of D5. Therefore, the formation and evolution mechanism of aggregate structure can be regarded as a process of the formation of strong and slightly strong contact forces and the transformation from slightly strong contact force to strong contact force.a value, which indicates that the particle roughness can increase the degree of interlocking of the aggregate. In addition, P(f) and iN of the strong contact force and iN of the slightly strong contact force increase with the increase in a value, but P(f) of the slightly strong and weak contact force decreases with the increase in a value. This shows that the increase in the aggregate roughness promotes the transformation from slightly strong and weak contact forces to strong contact force and slightly strong contact force, thus increasing the interlocking forces of the aggregate structure.a value and optimal mass percentage, respectively.As shown in a values of 0.4, 0.6 and 0.8 are used in the calculation of coarse aggregate gradation. Hence, the calculation table of coarse aggregate is shown in The ratio of CF aggregate  is determined to be 90:10, 85:15, 80:20, 75:25, 70:30, 65:35 and 60:40. It should be noted that since real aggregate is generally rarely round or round-like, the The performance of the asphalt mixture with different CF ratios is shown in According to The grading range determined based on the previous test results can be seen in Based on In order to compare the performance indicators of SG and RG, the results of the rutting test(dynamic stability) and SCB test(fracture toughness) are used to detect the high and low temperature performance of the asphalt mixture, as shown in As shown in Although some useful conclusions and findings can be obtained from the numerical simulation study at the two-dimensional scale, the performance pattern of the asphalt mixture will definitely be affected by the three-dimensional scale, so the subsequent study will strive to expand from two-dimensional scale to three-dimensional scale.The coarse aggregate structure can be regarded as a contact force transmission system composed of some strong and sub-strong contact forces. Moreover, the formation–evolution mechanism of coarse aggregate structure can be regarded as a process of the formation of strong and sub-strong contact forces and the transformation from sub-strong contact force to strong contact force.The main body of coarse aggregate consists of 4.75~19 mm aggregate, which is composed of a relatively stable structural system and is less affected by other sizes of coarse aggregate. The 2.36~4.75 mm aggregate acts as the secondary structure of the aggregate structure body, which plays the role of supporting the main structure and, at the same time, induces the secondary strong contact force provided by 4.75~19 mm aggregate to transform it into strong contact force and provide additional secondary strong contact force.The weak contact force is mainly provided by the aggregates with the particle size less than 2.36 mm, indicating that this aggregate contributes slightly to the formation of coarse aggregate structural bodies. From the point of view of the contribution to the formation of the aggregate structure body, 2.36 mm can be used as the dividing point of coarse and fine aggregates.The strength of the aggregate structure increases with the increase in the roughness of the aggregate, which is due to the transformation of sub-strong and weak contact forces to strong and sub-strong contact forces.In the asphalt mixture with the recommended gradation, dynamic stability increases by 30%, and fracture toughness increases by 26%, when compared to the mixtures with the standard gradation.The effects of other aggregate (recycled aggregate) properties and different temperatures on the aggregates are considered in our subsequent studies.In this study, the mechanical properties of aggregate structures, pavement properties of asphalt mortar and the effect of CF ratio on material properties are studied by means of numerical simulation and laboratory test, respectively. A new gradation based on meso-structure optimization is formed and then verified by laboratory tests. The main conclusions are as follows:


To assess the individual and synergistic effects of 2-butyne-1,4-diol (BD) and chloride ions on the microstructure and residual stress of electrodeposited nickel, various nickel layers were prepared from sulfamate baths comprising varying concentrations of BD and chloride ions by applying direct-current electrodeposition. And their surface morphologies, microstructure, and residual stress were tested using SEM, XRD, EBSD, TEM, and AFM. While the nickel layers composed of pyramid morphology were prepared from additive-free baths, the surface flattened gradually as the BD concentration of the baths was increased, and the acicular grains in the deposits were replaced with <100> oriented columnar grains or <111> oriented nanograins; additionally, the residual tensile stress of the deposits increased. The addition of chloride ions to the baths containing BD significantly increased the residual stress in the nickel layers, although it only slightly promoted surface flattening and columnar grain coarsening. The effects of BD and chloride ions on the growth mode and residual stress of nickel deposits were explained via analysis of surface morphologies and microstructure. And the results indicate that the reduction of chloride ion concentration is a feasible way to reduce the residual stress of the nickel deposits when BD is included in the baths. Electrodeposition technology has been widely employed in many fields, such as coatings, electronics, and weapons, due to its ease of controlling the microstructure and properties of deposited materials by altering the electrolyte composition and electrodeposition parameters ,2,3,4. N4H6O2, BD), which neither contains N nor S elements, is a class-II brightener [2 changed successively from <110> to <100> and <111> with increasing BD concentration, and the surface morphologies became spiral-type and lozenge-type, while the actual growth plane was always (111). When the electrolytes were stirred, the addition of BD caused a smooth surface and bright appearance of the deposit [During the nickel electrodeposition, the addition of suitable organic additives in the electrolytes can significantly alter the grain size and texture of deposits to obtain the desired properties, such as hardness, toughness, and corrosion resistance ,6,13,14.ightener ,17. Its ightener , allowinightener ,18. Meanightener ,18. Therightener ,19. Alimightener  found th deposit ,21 but a deposit ,22.To reduce the residual tensile stress caused by BD, it is necessary to reduce the use of BD or to add a stress reliever to the electrolytes ,12. HoweThe primary aim of this study is to assess the effect of chloride ions on the action of BD, as well as their synergistic effects on electrodeposited nickel. For this, the evolution of the microstructure of nickel deposits with varying concentrations of BD and chloride ions was investigated by using complementary characterization methods. The residual stress and the Vickers hardness (HV) test were conducted to characterize the mechanical properties of the deposits. Thereafter, the growth process of the nickel electrodeposits was analyzed, and the mechanisms of BD and chloride ions affecting the residual stress and hardness were explained via analysis of surface morphologies and microstructure.3NH2)2•4H2O) was the solute supplying Ni2+, and boric acid (H3BO3) was the pH buffer. Furthermore, nickel chloride (NiCl2•6H2O) was employed as a chloride ion source and for solving the anode, and sodium dodecyl sulfate (C12H25SO4Na) served as the wetting agent. Nickel deposits were prepared from sulfamate baths via the direct-current electrodeposition technique. Three sulfamate baths  contained the same nickel ion concentration, but varying chloride ion concentrations were used. In these baths, nickel sulfamate , which was polished and cleaned before the deposition. The initial pH values of the baths were adjusted to 4.0 by adding sodium hydroxide or sulfamic acid solutions, and the temperatures of the baths were maintained at 323 K at a constant magnetic stirring rate of 800 rpm. All the nickel layers were deposited at a constant current density of 2 A/dm2. The electrodeposition was conducted for 2 h to reach a nominal thickness of ~50 μm according to Faraday’s law. The current efficiency was estimated by weighing the cathodes before and after deposition. Deposits with a thickness of 20 μm were also prepared to evaluate the thickness-based change in the residual stress.During the electrodeposition, sulfur-activated nickel (Ni content > 99.96 wt.% and S content 0.027–0.04 wt.%), which was specifically designed for the process, was used as a soluble anode to supplement the consumption of Ni(hkl) was calculated from the diffraction intensities of the (hkl) plane using the following equation [hkl and I0hkl are the diffraction intensities of the (hkl) planes in the nickel deposits and in nickel powder, respectively. The 4 major peaks, i.e., (111), (200), (220), and (311), were used as the total value of the denominator.The crystal structure of the nickel deposits was analyzed using X-ray diffraction  with Cu-Kα radiation λ = 0.15405 nm). To estimate the preferred orientation of the electrodeposited nickel, the relative texture coefficient RTC405 nm. TScanning electron microscopy  was used to examine the surface morphologies of the nickel deposits at an acceleration voltage of 5 kV and a working distance of 10 mm. Further, the electron back-scattered diffraction (EBSD) images of the nickel deposits along the growth direction (GD) were characterized via SEM at a step size of 0.07 μm. The data was processed using the Channel 5 software (Version: 5.0.9.0). The microstructures of some of the deposits were also investigated via transmission electron microscopy  operating at 200 kV. Additionally, an atomic force microscopy  was used to characterize the surface morphology in an area of 10 µm × 10 µm.2ψ method [2ψ vs. 2θ curves. Moreover, to evaluate the mechanical properties of the various electrodeposits, the Vickers micro-hardness measurement was conducted on the cross-section of the deposits, and a peak load of HV0.05 and a hold time of 10 s were employed for the measurements. The hardness values comprised the average of 6 test values.The residual stress in the nickel deposits was tested via non-destructive XRD techniques using the classical sinψ method . The peaThe surface morphologies of the nickel layers that were deposited from the sulfamate baths containing varying BD and chloride ion concentrations were characterized by SEM, as shown in After adding a small amount of BD (0.3 mmol/L) to the baths, the surface of the nickel deposits flattened b,f,j. ThBy increasing the BD concentration to 1 mmol/L, the surface morphologies of the deposits were further altered, and the synergistic effects of BD and chloride ions became evident. By further increasing the BD concentration to 3 mmol/L, the surface morphologies of the nickel deposits from the various baths became consistent again, and their surfaces were covered with tiny bumps, as shown in (hkl) of the nickel layers that were deposited from different baths. It is easy to find that the main diffraction peaks of the electrodeposits clearly matched with XRD peaks of the reference nickel phase (JCPDS: 00-004-0850). However, the peak exhibiting the highest intensity varied with the BD concentration. In the absence of the additive, the (220) peak exhibited the highest intensity, and its RTC(220) value was >84%, indicating the presence of a strong <110> texture in the nickel deposits. The formation of the <110> texture in the nickel layers was attributed to the inhibition of the surface-adsorbed hydrogen atoms (Hads) [(220) value decreased slightly with increasing chloride ion concentration in the baths, indicating a decrease in the amount of Hads on the surface. After adding 0.3 mmol/L BD to the baths, only the (200) peak appeared distinctly and other peaks disappeared almost completely. RTC(200) increased to ~100%, and the deposits exhibited a strong <100> texture. Further, increasing the BD concentration to 1 mmol/L slightly altered the RTC(200) value, and the <100> texture in the deposits was unaffected. The <100> texture corresponds to the free lateral crystal growth mode and is generally associated with coarse grains and low residual stress [2 precipitation on the deposit surface resulted in the formation of this texture [2 precipitation on the surface. Put simply, the preferred orientation changed successively from <110> to <100> and <111> with the increase in the BD concentration, and the chloride ions hardly impacted it.s (Hads) . Additiol stress . The add texture . FurtherEBSD analysis was performed to study the microstructures of the electrodeposits; the inverse pole figure (IPF) maps of the cross-section of the nickel layers are shown in After adding 3 mmol/L BD in the baths, the grain size in the deposits was reduced significantly, lower than the resolution of EBSD. Therefore, TEM was performed to ensure a clearer observation of the microstructure. 2ψ method. And the deposits with thicknesses of 20 µm and 50 µm were tested to investigate the variation in the residual stress based on the thickness of the deposits, and the results are shown in The growth of the electrodeposits followed a layer-by-layer process, which generally forms residual stress in the deposits. The residual stress of the deposits was determined via XRD using the sinHardness is a crucial property of electrodeposited nickel . Figure 2), the reduction of Ni2+ was accompanied by that of H+; thus, the cathode surfaces were covered by a large amount of Hads, which promoted the formation of the <110> texture [+, which exhibited a low concentration in the acidic baths (pH = 4) [+ reduction, which facilitated the alkalization of the solution near the electrode surface and the formation of NiOH+ [ads adsorbed on the surface, imparting Deposit A-0 with a slightly strengthened <110> texture.During nickel electrodeposition, the chemical species in the baths can be adsorbed onto the cathode surface owing to the high surface energy of nickel, and the adsorbed species may significantly affect the nickel deposition process, resulting in changes in the morphology, microstructure, and properties of the deposits . Regardi texture ,32. The  texture . In addi(pH = 4) ,36. At tof NiOH+ ,37. The When BD was added to the sulfamate baths, it acted as a strong inhibitor for nickel deposition; its reaction kinetics were controlled by the mass-transport process ,38,39. Dads on the surface and caused the free-growth mode to operate. However, the BD molecules adsorbed on the cathode surface and inhibited the deposition of nickel atoms on the sites they occupied. The inhibition strength of BD was related to its coverage on the surface, and they were first adsorbed on (100) planes on the apex of the bumps via diffusion [ads or hydrogen gas at the grain boundaries. For nickel layers (Deposits A-0.3 and A-1) electrodeposited from the baths without chloride ions, the hydrogen reduction, which occurred prior to the nickel deposition [+ near the apex of the peaks and benefitted the out-growth of the micropeaks. Hence, in When the BD concentration in the baths was below 3 mmol/L, the deposits developed a <100> texture, the surface was flattened, and the coarse columnar grains appeared and increased in number with increasing BD concentration. The transformation of the texture from <110> to <100> can be explained by the hydrogenation of BD molecules, which rapidly consumed Hiffusion . The higiffusion . The forposition ,41, may 2 precipitation. Ni(OH)2 is considered a very efficient inhibitor of nickel growth, as well as a stabilizer of the <111> texture [2, the growth of the grain stopped frequently, and the deposits grew via repeated nucleation. Therefore, the grains in the deposits were fine, and the surface was flat. The growth type of these deposits corresponded to the unoriented dispersion type (UD) [After adding 3 mmol/L BD to the baths, the electrodeposited nickel layers were composed of nanoscale grains and formed a <111> texture, and their surface exhibited a large number of tiny bumps. During the electrodeposition, the hydrogenation of excessive BD molecules consumed extra hydrogen, which caused a rapid increase in the pH near the electrode surface, favoring Ni(OH) texture ,32. Undeype (UD) . The hig+Cl− ion pairs, which were readily included in the deposits due to they were electrically neutral. The subsequent desorption of OH− and Cl− may lead to the shrinkage of the deposits and the formation of residual tensile stress. The higher the chloride ion concentration in the baths, the more the ion pairs would be formed and incorporated in the deposits, and the greater the residual tensile stress of the deposits would be. The hydrogenation of BD promoted the alkalization of the solution near the electrode surface, which may favor the formation of the ion pairs; thus, BD in the baths enhanced the effect of chloride ions on the residual stress.The residual stress in the electrodeposited nickel was affected by numerous factors ,43. The 2 on the surface was partly incorporated into the deposits, and the deposit shrinkage, which was caused by the desorption of OH-, resulted in the formation of high tensile stress [Further, adding BD to the baths can generally induce an increase in residual tensile stress in the deposits. When the BD concentration was below 3 mmol/L, a strong <100> texture was formed in the deposits. Additionally, the adsorption and inhibition of BD first occurred on the (100) planes , which we stress . The dece stress ; thus, Ad. The deviation of partial points may be due to the influences of defect density, texture, and grain boundary structure [2 adsorption resulted in the increased interference of the deposition of the active sites, forming additional defects and further reducing the crystal size, and continuously increasing the hardness.It is well known that the hardness of materials is closely related to their grain size and that a Hall–Petch relationship exists between them ,47. The tructure . AlthougThe addition of BD to the sulfamate baths resulted in the replacement of the <110> oriented acicular grains by the <100> oriented coarse columnar grains or <111> oriented nanograins, as well as the increases in the residual stress;For BD-containing baths, BD molecules were adsorbed on the cathode surface and inhibited nickel deposition. And the grains in the deposits grew through the continuous formation and merging of the micropeaks on the surface. These processes determined the surface morphologies and microstructure, therefore affecting the residual stress and hardness of the deposits;Adding chloride ions into the baths could improve the inhibition of BD, which slightly promoted surface flattening and the <100> oriented grain coarsening, but evidently increased the residual stress in the nickel layers. The BD enhanced the effect of chloride ions on the residual stress. Therefore, reducing the concentration of chloride ions in the baths is a feasible way to obtain a low residual stress nickel deposit when using BD-containing baths for electrodeposition.Herein, the microstructure and residual stress of electrodeposited nickel layers, which were prepared from the sulfamate baths containing varying BD and chloride ion concentrations, were tested. The individual and synergistic effects of BD and chloride ions were explained via analysis of the results. The key conclusions are as follows:


Di-isononyl phthalates are chemicals that are widely used as plasticizers. Humans are extensively exposed to these compounds by dietary intake, through inhalation and skin absorption. Sulfotransferases (SULTs) are enzymes responsible for the detoxification and elimination of numerous endogenous and exogenous molecules from the body. Consequently, SULTs are involved in regulating the biological activity of various hormones and neurotransmitters. The present study considers a computational approach to predict the toxicological potential of the metabolites of di-isononyl phthalate. Furthermore, molecular docking was considered to evaluate the inhibitory potential of these metabolites against the members of family 1 of SULTs. The metabolites of di-isononyl phthalate reveal a potency to cause liver damage and to inhibit receptors activated by peroxisome proliferators. These metabolites are also usually able to inhibit the activity of the members of family 1 of SULTs, except for SULT1A3 and SULT1B1. The outcomes of this study are important for an enhanced understanding of the risk of human exposure to di-isononyl phthalates.  Xenobiotics are chemical compounds that do not occur naturally in the human body and, if not metabolized, they could reach toxic concentrations. Humans are exposed daily to numerous xenobiotics: drugs, food additives, cosmetic ingredients, pesticides, environmental pollutants, etc. The metabolism of xenobiotics usually occurs through enzymatic reactions, and the enzymes involved are classified into phase I, phase II, and transporter enzymes. Phase I enzymes (such as cytochromes P450) convert lipophilic xenobiotics into more polar compounds for easier excretion and provide sites for conjugation reactions. Phase II enzymes (such as glutathione transferases and sulfotransferases) carry out the conjugation reactions and can interact directly with xenobiotics, but most of the time they interact with the metabolites obtained in the phase I metabolism . This study focuses on family 1 of sulfotransferases (SULT1), containing enzymes that are known to catalyze the sulfate conjugation of pharmacologically important endobiotics and xenobiotics by transferring a sulfate group from the donor 3′-phosphoadenosine 5′-phosphosulfate (PAPS) to the hydroxyl group of an acceptor . In humaSeveral polymorphisms have been identified for the SULT1 family of enzymes that proved to alter enzyme activities and presented differences in the metabolism of endogenous compounds, drugs and other xenobiotics ,5. The fAmong other xenobiotics, humans are exposed to phthalates because these compounds are widely used as solvents, additives and/or plasticizers in numerous consumer products. People can be exposed to phthalates by ingesting them from food or drink, by inhaling phthalate-contaminated air and dust resulting from paintings or furniture, by using pharmaceuticals and cosmetics, and by skin contact with products and/or clothing that contain them ,7. ThereIn humans, the metabolism of phthalates occurs in at least two steps: (i) a phase I hydrolysis when the diester phthalate is hydrolyzed into the monoester phthalate and (ii) a phase II conjugation . Some inhttps://echa.europa.eu/documents/10162/31b4067e-de40-4044-93e8-9c9ff1960715, accessed on 15 June 2023). Investigations conducted on rats and on human adult volunteers have shown that DiNP has been metabolized to a number of secondary metabolites before being excreted in the urine: monoisononyl phthalate (MiNP), mono-hydroxy-isononyl phthalate (MHiNP), mono-carboxy-isooctyl phthalate (MCiOP) and mono-oxo-isononyl phthalate (MOiNP) [Di-isononyl phthalates (DiNP) are a group of isomeric forms of dinonyl phthalates widely used as plasticizers and found in various products such as cables, foil, PVC flooring, toys, shoe materials, tablecloths, etc. . It was  (MOiNP) ,19,20. This study aims to predict the possible toxicological effects on humans of the metabolites of DiNP and to assess the interactions of these xenobiotics with members of the SULT1 family, also taking into account the most frequently identified polymorphic variants of SULT1A1 enzymes. To obtain this information, a computational study was implemented.Sometimes the reliability of the results obtained by using calculation methods is called into question. Considering both the cost of laboratory studies and the ethical concerns of using animals for testing, the role of computational studies is becoming well recognized. There has been an explosive growth in both the scale and diversity of data in the natural sciences, leading to the creation of freely accessible databases and enabling scientists to build accurate computational models for toxicology assessments. Consequently, the Organization for Economic Development and Cooperation (OECD) created guidelines for quantitative structure activity relationship (QSAR) models in 2004. The principles for building QSAR models and calculation methods have been described in detail since 2007 . The RegThe physicochemical and structural properties of the DiNP metabolites and of the ligands that are present in the crystallographic structures of SULT1 enzymes are presented in Data presented in Considering the fact that DiNP metabolites were observed in urine from human subjects, predictions about their toxicity on humans have been also obtained by using the ADMETLab2.0 computational tool ,24. ThisThe predicted toxicological effects of DiNP and of its metabolites on the human organism are illustrated in The following can be observed from The outcomes of the molecular docking are revealed in In the case of the allelic variant SULT1A1*2, MiNP and MOiNP are the DiNP metabolites that are able to bind to the active site of the enzyme .As a control, the molecular docking of p-nitrophenol  to SULT1A1*2 emphasizes that the binding mode of p-nitrophenol obtained through molecular docking strongly corresponds to the position of one of the p-nitrophenol molecules in the crystallographic structure, the binding energy being −6.05 kcal/mol . The binAll the DiNP metabolites are able to bind to the active site of SULT1A1*3 .In the case of SULT1A2, MCiOP, MiNP and MOiNP are able to bind to the active site of SULT1A2 . MHiNP dNone of the DiNP metabolites bind to SULT1A3 and SULT1B1 active sites .In order to assess the binding of DiNP metabolites to SULT1C1, first the molecular docking of iodothyronine  to the enzyme has been completed. The binding to the active site of SULT1C1 has been emphasized by superposition of the binding modes of iodothyronine to SULT1C1 with the structures of SULT1A1*2 and SULT1B1 . FurtherAll the DiNP metabolites are able to bind to the active site of SULT1C1  and the All the DiNP metabolites are also able to bind to the active site of SULT1E1 , with thThe energies resulting from the molecular docking study and corresponding to the binding modes matching the active sites of SULT1 enzymes are presented in Data presented in The results obtained using PLIP 2021 software regarding the identification of the residues of the SULT1 enzymes that are involved in the noncovalent interactions with DiNP metabolites are presented in Information presented in To the best of our knowledge, this is the first study dealing with the possible toxicity of DiNP and its metabolites. Predictions obtained using the ADMETLab2.0 tool revealed that DiNp may be an inhibitor of P-glycoprotein, underlining that DiNP may potentially affect the absorption, distribution, metabolism and elimination of the substrates of this protein. The high degree of binding to plasma proteins of the DiNP and its metabolites may lead to long acting xenobiotics as their bound fractions are not available for metabolism or excretion. The present study emphasizes that the DiNP metabolites may produce liver damage and this outcome is in good correlation with literature data revealing that several phthalate metabolites produced liver injuries . The potNone of the DiNP metabolites are considered to produce cardiotoxicity, mutagenicity or carcinogenicity. These predictions correlate with no carcinogenic  and mutaSeveral toxicological effects on humans have been predicted for numerous other xenobiotics ,31,32,33A molecular docking study reveals that MCiOP and MiNP bind to the active site of SULT1A1*1, but MHiNP and MOiNP do not bind to the active site of this enzyme. This distinct behavior of MCiOP and MiNP that binds to SULT1A1*1 may be associated with their higher molecular lipophilicity potential  benzene-1,2-dicarboxylate) are considered in the present study : mono-cahttps://www.molinspiration.com/cgi/galaxy, accessed on 4 June 2023). For comparison purposes, the physicochemical and structural properties of ligands that are present in the crystallographic structures of SULT1 enzymes have been also obtained. Physicochemical properties and three-dimensional structures of the metabolites of DiNP have been extracted from PubChem database . Surface2 > 0.72. There are numerous other computational tools that can be used to assess the ADMET properties, with ADMETLab2.0 tool being used in this study because it is based on well-performed prediction models and has been used by more than 50,000 users in recent years, with millions of compounds screened [ADMETLab2.0 server has been used in order to obtain information regarding the possible human organ toxicological effects of DiNP and of its metabolites. The classification models used for predicting the toxicity endpoints have a minimum accuracy of 80%; regarding the regression models, most of them achieved an Rscreened ,24. The The molecular docking approach has been considered for assessing the possible interactions of the DiNP metabolites with the SULT1 family. In order to implement the molecular docking study, the structures of enzymes and those of the metabolites of di-isononyl phthalates are necessary. Structures of DiNP metabolites were extracted from PubChem database. Structures of SULT1 enzymes have been extracted from Protein Data Bank and are described in It is known that both SULT1A1 and SULT1A2 have p-nitrophenol as a specific substrate; in order to interpret the docking outputs, structures of the SULT1A2 and of the variants SULT1A1*1 and SULT1A1*3  have been superimposed to the structure of SULT1A1*2 that corresponds to the complex formed by the SULT1A1*2 variant with the inactive cofactor adenosine-3′-5′-diphosphate (A3P) and two molecules of p-nitrophenol . The supIn the case of SULT1C1, in order to locate the catalytic cavity and to more easily interpret the molecular docking outcomes, its crystal structure has been superposed with both that of SULT1A1*2 containing the inactive cofactor A3P and two molecules of p-nitrophenol, and that of SULT1B1 that contains the substrate resveratrol, with the superposition also reflecting a high similarity between the three structures . FurtherThe noncovalent interactions between the DiNP metabolites and the SULT1 enzymes have been analyzed using the Protein–Ligand Interaction Profiler tool .The results obtained in the present study reveal several human health risks of the metabolites of di-isononyl phthalate. These compounds revealed the capacity to cause liver damage, to inhibit receptors activated by peroxisome proliferators and are usually able to inhibit the activity of family 1 of sulfotransferases, except SULT1A3 and SULT1B1. Taking into account that humans are extensively exposed to phthalates and the important role of sulfotransferases for the detoxification and elimination of numerous endogenous and exogenous molecules in the body, information regarding the biological effects of the metabolites of phthalates and their possible inhibitory effects against the activities of sulfotransferases contributes to better understanding the risk of human exposure to phthalates.


Injection molding (IM) is already an established technology for manufacturing polymer products. However, in the course of the increased use of recyclates for economic and ecological reasons, its application capability has been confronted with new requirements for reliability and reproducibility. In addition, the IM process is confronted with regulations regarding a verifiable recycling degree in polymers. With regard to the material identification and storage of manufacturer-, process- or product-related data in polymers, the implementation of a material-inherent marking technology forms a potential answer. The IM process combined with modified polymers (MP) as a marking technology turns out to be a feasible approach to manufacturing reproducibly and offers a high quality based on increased process awareness and fulfilling the required traceability. Therefore, this work focuses on the trial evaluation of MP within the IM process. The influence of MP on the material process behavior and mechanical and thermal component properties, as well as the influence of the IM process and recycling on MP traceability, are investigated. No discernible influences of MP on the investigated properties could be identified, and the traceability from the initial material to a recyclate could be confirmed. MP is suitable for monitoring the aging state of polymers in IM. Injection molding (IM) is among the most prominent representatives of polymer-based manufacturing processes and is characterized by a causal flow from the initial material to the end product. In the IM process, the plastification of granular or powdered polymers into a flowable state occurs using dissipation and heat conduction along a temperature-controlled cylinder. The plastified material is injected into the closed mold via the axial movement of the injection unit and subsequently solidifies. In the case of thermoplastic materials, the mold is temperature-controlled to a temperature below the solidification temperature of the melt so that the melt cools and solidifies as it enters the mold. Part quality is influenced by the IM process parameters, such as the temperatures from the hopper to the nozzle, the holding pressure and the back pressure, as well as the screw rotation speed, holding time and mold temperature ,2,3,4.The widespread use of thermoplastics and their composites in the present is a cause of growing concern because of their adverse effects on the environment . AlthougAs different batches and material streams have different material properties, the identification of the different classes of recyclates is required ,32,36,37IM combined with modified polymers as a marking technology turns out to be a promising way to manufacture reproducibly and with high quality, based on an advanced process awareness . TherefoConsequently, the aim of the present study is to validate modified polymers for application in IM. Investigations are carried out to determine the influence of the modified polymer on the processing behavior of the material and the component properties and to determine the influence of the IM process and recycling on the traceability of the modified polymer. Thermal and rheological material properties, as well as mechanical and thermal component properties, are analyzed. Since repeated extrusion and IM are evaluated as suitable methods to investigate recyclability ,44,45,46In order to minimize disturbance variables and avoid aging processes due to humidity, temperature and ultraviolet radiation ,14,21,483. The material choice was based on the fact that polypropylene is one of the most commonly used semi-crystalline thermoplastics in injection molding [The Daplen EF155AE elastomer-modified polypropylene compound with 10% mineral filling from Borealis AG  was used. This material is among the thermoplastic polyolefins ,15,49 an molding ,50,51,52 molding .® technology [The modified polymer used in this study was supplied by Polysecure GmbH  and branded as POLTAGchnology ,54,55,56chnology  listed fchnology , this moOn the basis of the used injection molding material and modified polymer, a master batch was produced. Via a spray-drying process, the modified polymer was added to the injection molding material via mixing. In the master batch, a concentration of 30 ppm of the modified polymer was used. The method of producing the master batch based on the modified polymer was confirmed in a previous study . ThroughIn In Then, the ground material was extruded by a twin screw extruder of the ZE25A-40D-UTX-UG type from KraussMaffei Berstorff  with gravimetric dosing from Scholz Dosiertechnik GmbH . All materials were extruded according to the parameters listed in Before the next cycle (R1), a granulate quantity of 20 g was taken for further tests. The remaining amount of granulates was further processed. Then, the pre-dried granulates were injection-molded, and the recycling process was repeated up to ten times (R10). The number of recycling steps was based on Aurrekoetxea et al. , and it Thermogravimetric analysis (TGA) was applied using a TGA/DSC-1 measuring system from Mettler Toledo . Measurements were performed according to the DIN EN ISO 11358-1:2022-07  standard+ measuring system from Mettler Toledo . Measurements were obtained according to the DIN EN ISO 11357-1:2023-06 [Differential scanning calorimetry (DSC) was performed using a DSC-3:2023-06  standardMelt flow index testing was realized with an Mflow measuring device from Zwick/Roell GmbH & Co. KG  and was performed according to the DIN EN ISO 1133-1:2012-03  standardA tensile test was performed on a tensile testing machine of the Zwick/Roell Z100 type from Zwick/Roell GmbH&Co. KG. The equipment of the testing machine included a makroXtens mechanical extensometer and two wedge clamping jaws designed for a normal force of up to 10 kN. Furthermore, the equipment included an Xforce K load cell determined for the same load limit and a testControl II control unit. According to the DIN EN ISO 19069-2:2020-01  standardA Charpy impact test was performed on a pendulum impact tester of the Zwick/Roell HIT25P type from Zwick/Roell GmbH & Co. KG using a testControl II control unit. According to the DIN EN ISO 179-1:2010-11  standard2. The measured temperature was defined as the VST. Three specimens were used for each measurement. The mean value and the standard deviation were used to determine the VST. The objective is to quantitatively characterize the thermal stability of a polymer [The Vicat softening temperature (VST) of the specimens was measured according to the DIN EN ISO 306:2023-03  standard polymer . Since t polymer ,69.Tandem mass spectroscopy (MS/MS) was used to detect, sequence and measure the traceability of the modified polymer ,70,71,72Based on a Python tool used in a previous study , the resIn M) between the first and fifth steps for all concentrations considered, there was a decrease in TM from R0 to R10 for all concentrations observed. From R0 to R10, the TM for 0 ppm decreased by approximately 0.36%. In contrast, for 1 ppm, the TM decreased by approximately 0.09% from R0 to R10, and for 30 ppm, it decreased by approximately 1.31%. At R0, the deviations of the TM between the unmarked material and the marked materials amounted to a maximum of approximately 0.34%. At R1, the maximum deviation of TM was approximately 0.8%. At R10, the deviations of the TM between the unmarked material and the marked materials amounted to a maximum of approximately 1.47%.The influence of the modified polymer on the thermal process window, characterized by melting and crystallization temperature and depending on the concentration of the modified polymer and the recycling step, is presented in C), there was an increase from R0 to R10 for nearly all concentrations considered. From R0 to R10, the TC for 0 ppm increased by approximately 0.53%. For 1 ppm, the TC increased by approximately 1.54% from R0 to R10, and for 30 ppm, it increased by approximately 1.42%. At R0, the deviations of the TC between the unmarked material and the marked materials amounted to a maximum of approximately 0.32%. At R1, the maximum deviation of TC was approximately 1.17%. At R10, the deviations of the TC between the unmarked material and the marked materials amounted to a maximum of approximately 0.96%.Regarding crystallization temperature , depending on the concentration of the modified polymer and the recycling step, is presented in The mean value of the MFR for 0 ppm increased by approximately 4.61% from R0 to R1. In contrast, for 1 ppm, the mean value of the MFR increased by approximately 4.08% from R0 to R1, and for 30 ppm, it increased by approximately 4.92%. From R1 to R10, the mean value of the MFR for 0 ppm increased by approximately 0.72%. In contrast, for 1 ppm, the mean value of the MFR decreased by approximately 3.43% from R0 to R10, and for 30 ppm, it increased by approximately 3.40%. At R0, the deviations of the mean value of the MFR between the unmarked material and the marked materials amounted to a maximum of approximately 2.97%. At R1, the maximum deviation of the mean value of the MFR was approximately 2.45%. At R10, the deviations of the mean value of the MFR between the unmarked material and the marked materials amounted to a maximum of approximately 2.55%. The deviations of the mean values of the MFR as a function of the concentration of the modified polymer were also within the standard deviation across the recycling steps.The influence of the modified polymer used on the mechanical properties, depending on the concentration of the modified polymer and the recycling step, is presented in The influence of the modified polymer on the double-edge-notched impact strength (dNIS), depending on the concentration of the modified polymer and the recycling step, is presented in The mean value of the dNIS for 0 ppm increased by approximately 5.13% from R0 to R10. In contrast, for 1 ppm, the mean value of the dNIS increased by approximately 4.33% from R0 to R10, and for 30 ppm, it increased by approximately 5.52%. At R0, the maximum deviation of the mean value of the dNIS between the unmarked material and marked materials was approximately 2.63%. At R10, the maximum deviation of the mean value of the dNIS between the unmarked material and marked materials was approximately 3.38%. The deviations of the mean values of the dNIS, depending on the concentration of the modified polymer, were within the standard deviation even across the recycling steps, with the exception of recycling step R1.The influence of the modified polymer on the Vicat softening temperature (VST), depending on the concentration of the modified polymer and the recycling step, is presented in The maximum deviation of the mean value of the VST between the unmarked material and marked materials at R0 was approximately 1.06%. At R10, the maximum deviation of the mean value of the VST between the unmarked material and marked materials was approximately 0.59%. The deviations of the mean values of the VST as a function of the concentration of the modified polymer were also within the standard deviation across the recycling steps.The traceability of the modified polymer at different recycling steps is presented in The determined intensities did not increase linearly with the selected concentrations. Furthermore, there was no apparent correlation between the concentration of the modified polymer and the mean value of the detected intensity of the main mass . If the The investigated thermal and rheological material properties , as wellThe selected concentrations of the marking agent are in accordance with an earlier study . In addiAs there is no apparent correlation between the investigated concentration of the modified polymer and the determined mean value of the intensity of the main mass, only a qualitative analysis is available. This observation is confirmed by a previous study . AdditivRegarding the melting temperature, a small decrease occurred from R0 to R10 for all concentrations . This isThe investigated increasing mean value of the melt flow rate (MFR) from R0 to R10, illustrated in The resulting investigated mechanical component properties at R0 are in accordance with previous studies ,87. WithWith regard to tensile strength , there aRegarding elongation at break , there aAlthough no clear increase in Young’s modulus or tensile strength could be observed, as indicated by Aurrekoetxea et al. , the incRegarding Charpy impact strength , there aWith regard to the Vicat softening temperature (VST) , there aThe identification and traceability of materials and components is a major challenge in the injection molding process. This challenge can be overcome through the use of a material-inherent marking technology. This allows the aging state of polymers and the composition of individual material mixtures to be monitored, as a result of which scatter-reducing process parameters can be set. Based on a systematic selection of modified polymers as a proper marking technology for application in polymer processing manufacturing, as well as the experimental validation of this marking agent in selective laser sintering in previous studies ,39, thisConsidering the applied investigations and concentrations of the modified polymer used in the injection molding material used, the marking technology does not have any discernible influence on the investigated properties of the material and components;During injection molding, the marked material shows analogous processing behavior to the unmarked material;The modified polymer used can be reliably detected in the material down to a concentration of 1 ppm, even after ten recycling steps;The concentration of the modified polymer and the measured intensity of the marking agent in the injection molding material used do not correlate;The dispersion of the modified polymer used in the injection molding material is sufficient;The modified polymer is suitable as a marking agent in injection molding;The modified polymer allows encoding of the material used at the molecular level.The applicability of modified polymers for use in injection molding can be confirmed. This demonstrates the thermal stability of modified polymers ,55 for aBased on this study’s findings, an injection-molding-specific coding strategy can be developed. The tracer-based process optimization  must be


Cluster molecular magnets prove their potential for applications in quantum technologies, encouraging studies of quantum entanglement in spin systems. In the paper we discuss quantum entanglement properties of pentamer cluster composed of spins  Molecular magnetism constitutes a flourishing field of study within the theoretical and experimental condensed matter physics and chemistry ,2, combiThe interest in quantum entanglement brings the localized spin systems of cluster geometry to the attention of the theoreticians ,22,23,24Among various shapes of clusters, a pentamer geometry can be highlighted. Such clusters attracted so far some attention of theorists, but only in the context unrelated to quantum entanglement, to mention the neutron scattering simulation ,68, magnS=1/2 . This paS=1/2 . TherefoMotivated by the synthesability of pentamer spin clusters described above in the field of molecular magnetism and by their non-trivial, highly symmetric geometry, we present a theoretical study focused on characterization of quantum entanglement in such structure. In order to capture the maximized quantum effects, we select cluster composed of spins The system of interest is a spin pentamer cluster, composed of spins An interesting feature of the studied cluster is that it forms a smallest nonplanar Kuratowski graph, being a complete graph of 5 vertices K5 , each veentamers ,81.We model the system with isotropic Heisenberg Hamiltonian:i, moreover, H.In the above Hamiltonian, the symbol z axis It should be noticed that the Hamiltonian  commutesHowever, it must be stated that some of the states labelled with three quantum numbers The symbolic or numerically exact diagonalization of the Hamiltonian matrix provides a set of eigenvectors In the description of system states in our work, the single-spin states with spin up (down) are marked with Out of Bell states, the state H. The thermodynamic description of the system is constructed following the rules of canonical ensemble of statistical physics. Let us put emphasis on the fact that the external magnetic field is directly included in the Hamiltonian  rather than pure states. Therefore, if we deal with s. The remark about mixed ground state also applies to the case of the phase boundary  or a triple/quadruple point (where three/four ground states indicate the same energy) when the degeneracy appears necessarily, even if the neighbouring ground states are pure ones. In both such cases the state of the system is a mixed quantum state being a probabilistic mixture of all the involved Hamiltonian eigenstates sharing the same energy along specific line or at point . In the absence of the external magnetic field, for s share the same energy.If the case of Equation ] and alls for spin of tetrahedron). In the presence of the external magnetic field s. On the contrary, in the case of a mixed state, it is described by a density matrix s).If the ground state for a given range of parameters is non-degenerate, it is marked in In both panels of The ground-state phase diagram for ferromagnetic /2,2 see , having For Equation  involvinIn this state, the central spin inside the tetrahedron (labelled with 1) takes the pure state For Equation , composeAlso in this state, the central spin inside the tetrahedron (labelled with 1) takes the pure state Let us separately mention the form of the ground states exactly at If Equation  with d=6If the coupling fulfils the condition Equation  with theSome of the above states, corresponding to Finally, if Equation  with d=4The ground-state phase diagram for antiferromagnetic stem see ; it has In this state, the central spin inside a tetrahedron, labelled with 1, is in mixed state of the form If Equation ] composeIn this state, the spin labelled with 1 is in mixed state of the form Again, for If Equation ] containIn the case of Equation ] is formFinally, if Equation  with d=4After analysis of the ground-state phase diagram it can be concluded that relatively large area is filled with states showing degeneracy, due to high symmetry of the studied cluster.The system ground states can exhibit entanglement for the case of ferromagnetic J1 a and of J1  spins are less entangled, with concurrence value of 1/10 and quantum state:Let us note that also the boundary of both spin-3/2 states shows the presence of entanglement, as well as the triple point at which two spin-3/2 states and spin-5/2 state have the same energy. Also the boundary between spin-3/2 and saturated ferromagnetic state corresponds to non-vanishing entanglement of both kinds of spin pairs. On the other hand, at the boundary between the pure spin-3/2 state and spin-1/2 state The effect of the finite temperature Equation ]. In thettention ,91,92 anttention ; noticeattention ). Again,The discussion of the finite-temperature properties will first involve the case of The effect of finite temperature on the concurrence values for The effect of variable temperature and magnetic field on the concurrence in the case of T increases , but its height is reduced when the magnetic field rises . The effect of the increasing magnetic field can be tracked in T) and a maximum is formed  composed of spins clusters . The intaterials . For theaterials  exposingThe system in question was inspired by synthesis of Cu-based cluster molecular magnets containing spins 1/2  and Co-bel study  or partiel study ,97).


Moreover, we noted a significant relationship between the serum profile of PTX3 and disease activity, expressed as CDAI, both before  and after  treatment. A similar correlation was noted in the case of S100A12 , albeit exclusively within the post-treatment group of patients. Anti-inflammatory treatment resulted in an elevation of pro-GN concentration  and a reduction in PTX3 level  in the serum of patients with CD. In comparison to our previous research conducted on a group of patients with ulcerative colitis (UC), those with CD exhibited reduced levels of PTX3  and elevated concentrations of S100A12 . The results obtained from this investigation suggest that measurements of pro-GN, PTX3 and S100A12 could prove beneficial in the diagnosis of Crohn’s disease. Assessment of changes in the serum profile of PTX3 appears to be a good marker of response to treatment but also, along with analysis of S100A12 protein serum levels, a useful marker in differentiating CD from UC.The aim of our case-control study was to identify novel biomarkers of Crohn’s disease (CD) that hold the potential to be employed in both disease diagnosis and monitoring activity. In the context of the contribution of intestinal barrier integrity and immune response to the pathogenesis of CD, we assessed the serum concentrations of proguanylin (pro-GN), pentraxin 3 (PTX3) and S100A12 in 20 patients before and after anti-inflammatory treatment, as well as in 20 healthy individuals. Statistical analyses revealed a significant difference in the levels of pro-GN (5.5 vs. 11.35,  Inflammatory bowel disease (IBD) is a chronic and inflammatory condition characterized by alternating episodes of clinical remission and relapse. It can be classified into two primary types of disorders—ulcerative colitis (UC) and Crohn’s disease (CD). In UC, the disease spreads proximally from the colon to other segments of intestines, affecting the mucosa and submucosa layers. In contrast, CD can lead to lesions anywhere along the gastrointestinal tract, extending through the entire tissue ,2. The oThe pathogenesis of Crohn’s disease involves multiple factors, but a crucial role is contributed to impaired intestinal barrier integrity and exceeded immune response . In our Other potential biomarkers of Crohn’s disease analyzed in this study are pentraxin 3 (PTX3) and s100A12 protein, both of whose function is related to the immune response. PTX3 is a glycoprotein belonging to the pentraxin family along with the well-known C-reactive protein (CRP). Similar to CRP, PTX3 plays a role in innate immune response, and its expression increases under inflammatory conditions. The synthesis of PTX3 occurs in various cell types, including monocytes/macrophages, epithelial cells, endothelial cells and smooth muscle cells. Neutrophils also serve as a cellular source of PTX3, storing the protein in their granules and releasing it immediately upon appropriate stimuli. The stimuli controlling synthesis and release of PTX3 include inflammatory cytokines, including IL-1β, TNF-α and microbial antigens like lipopolysaccharide (LPS) ,10. In tRegarding the contribution of proguanylin to intestinal barrier integrity and pentraxin 3 along with S100A12 to innate immunity, these mentioned biomarkers may be useful in the diagnosis of Crohn’s disease. Moreover, as the analyzed biomarkers are related to the inflammatory response, playing a key role in the progression of the disease, measurement of pro-GN, PTX3 and S100A12 might also be valuable in assessing disease activity and the intensity of the inflammatory process. Therefore, the hypothesis of our study is that pro-GN, PTX3 and S100A12 measurements in the serum of patients with Crohn’s disease are useful in diagnosing and monitoring the activity of the disease.The material investigated in this research consisted of venous blood collected from 40 adults including 20 healthy individuals and 20 patients with Crohn’s disease before treatment and after a year of anti-inflammatory therapy. The exclusion criteria for the control group were hospitalization or surgery within the year prior to study as well as pharmacological treatment in the prior month. Additionally, all the laboratory parameters assessed in the blood of healthy individuals had to be within the reference ranges. For the study group, the diagnosis of Crohn’s disease was made on the basis of clinical symptoms, endoscopic examination and laboratory tests at the Department of Gastroenterology of St. Barbara’s Regional Specialist Hospital in Sosnowiec. Moreover, the disease activity was evaluated using the Crohn’s Disease Activity Index (CDAI). The exclusion criteria among patients with CD were unstable coronary disease, myocardial failure, pregnancy or breastfeeding, diabetes, liver or kidney disease, severe viral, fungal or bacterial infections and precancerous or cancerous conditions. Patients included in this study were treated with prednisone, the dosage of which was adjusted for each patient individually and was gradually reduced up to 12 weeks. The anti-inflammatory treatment was further continued with azathioprine or mercaptiopurine.This research is a case-control observational study in which we compared two groups: a control group comprising healthy individuals and a study group including patients with a prior diagnosis of Crohn’s disease. The aim of this study was to identify the significance of changes in the levels of pro-GN, PTX3 and S100A12 in the serum of the subjects depending on the presence or absence of the disease. In addition, our research was designed as a prospective study; therefore, we continued the observation of the study groups in order to determine the progression of the disease during treatment with corticosteroids as well as to identify changes in the levels of analyzed biomarkers after one year of the mentioned therapy. The study size was determined based on the aforementioned inclusion criteria, and subjects included in the research were only newly diagnosed patients with no prior corticosteroid treatment.Serum proguanylin level was measured using the Human Proguanylin ELISA test from BioVendor Company . The analytical sensitivity of the test was 0.06 ng/mL, while the intra-essay variability was estimated at the level of 5.2%.Concentration of serum pentraxin 3 was assessed using a Human Pentraxin 3 ELISA test from BioVendor Company . The analytical sensitivity of the test equaled 22 pg/mL, while the intra-essay error was 3.4%.Assessment of the serum S100A12 concentration was determined with the use of a Human S100A12 ELISA test from BioVendor Company . The analytical sensitivity of the method was estimated at the level of 0.02 ng/mL, and the intra-essay variability was 3.6%.t test in normally distributed data and the Mann–Whitney U test in not normally distributed data. Comparison of results before and after the implemented treatment was conducted using Student’s t test and the Wilcoxon test. The correlation between the concentration of analyzed biomarkers and disease activity along with CRP were evaluated using the Pearson test. In all tests and statistical analyses, a significance level of p < 0.05 was used.Obtained data were characterized and evaluated with the use of STATISTICA software . The normality of data distribution was assessed using the Shapiro–Wilk test. In the case of normally distributed data, the descriptive statistics included mean values and standard deviation (SD), while not normally distributed data were characterized by the median (Me) and interquartile range—lower (Q1) and upper (Q3) quartiles. The statistical significance of differences between patients with Crohn’s disease and healthy individuals was assessed with the use of Student’s The clinical characteristics of patients with Crohn’s disease are presented in p < 0.001) by 52% compared to healthy individuals. At the same time, we noted a significant increase in PTX3 (p < 0.05) and S100A12 (p < 0.001) levels in the CD group by 32% and 302%, respectively, compared to control group.In this study, we aimed to evaluate the utility of pro-GN, PTX3 and S100A12 as diagnostic markers of Crohn’s disease. To achieve this, we assessed the levels of these mentioned parameters in the serum of patients with CD and healthy individuals, and the results are presented in p < 0.005, r = 0.63) and after  treatment. A similar correlation was identified in the case of S100A12  but only in the post-treatment group of patients with CD.Regarding the aim of this study to identify new biomarkers reflecting disease activity, we evaluated the correlation between serum profiles of pro-GN, PTX3 and S100A12 and the CDAI score. The correlations between CDAI score and the analyzed biomarkers are presented in Moreover, as the median CRP concentration in the blood of patients with Crohn’s disease was increased compared to the reference values, we estimated the correlation between the level of this inflammatory marker and serum profiles of pro-GN, PTX3 and S100A12. Unfortunately, statistical analyses revealed no significant relationship between the concentration of the analyzed biomarkers and CRP in the pre- and post-treatment groups of patients with CD.p < 0.001) and PTX3 (p < 0.05). The increase in serum pro-GN concentration reached 46%; however, in comparison to healthy individuals, the level of pro-GN remained decreased (p < 0.005). After one year of anti-inflammatory treatment, we observed a decrease in PTX3 levels by 24%, reaching values similar to the control group. However, no significant change was observed in the case of S100A12 after therapy. The level of S100A12 in the post-treatment group of patients with CD remained significantly increased compared to healthy individuals , while the level of S100A12 (p < 0.05) was increased compared to the subjects with ulcerative colitis before treatment. On the other hand, after implemented treatment, the serum profiles of pro-GN, PTX3 and S100A12 did not differ significantly between UC and CD patients.In this study, we aimed to evaluate the use of serum pro-GN, PTX3 and S100A12 measurements in the differential diagnosis of inflammatory bowel disease. To achieve this, we referred to our previous research, conducted in the Department of Clinical Chemistry and Laboratory Diagnostics , in whicThe pathogenesis of Crohn’s disease is linked to disrupted intestinal barrier integrity, resulting in the activation of the immune response and the subsequent initiation and development of inflammatory process within the intestines . As impaIn this study, we also assessed the concentration of pentraxin 3 and S100A12, which are proteins related to the innate immune response and are engaged in the development of inflammation, being an integral part of the pathogenic process during Crohn’s disease. The conducted statistical analyses revealed a significant difference in the concentration of both PTX3 and S100A12 between patients with CD and healthy individuals. The levels of PTX3 and S100A12 in the serum of CD patients were markedly increased. A similar increase in PTX3 expression was also reported in the study performed by Chen et al. , where PRegarding the crucial relationship between the analyzed biomarkers and the pathogenic process during Crohn’s disease, we also assessed the usefulness of serum pro-GN, PTX3 and S100A12 measurements in evaluating disease activity. The conducted statistical analyses revealed no significant relationship between pro-GN concentration and the CDAI score. However, we noted a significant correlation between the profile of PTX3 and disease activity both before and after anti-inflammatory treatment. Corresponding results were presented in Chen et al.’s study , where aAnother significant correlation in this study was found between the level of S100A12 protein and the CDAI score in the group of patients with CD, but only after anti-inflammatory treatment. Similar to the results obtained in our study, Däbritz et al.  also notIn this study, we analyzed the levels of pro-GN, PTX3 and S100A12 in the serum of patients before and after one year of anti-inflammatory therapy. Implemented treatment significantly decreased the activity of the disease in patients with CD. Moreover, in the post-treatment group of patients with CD, we noted a significant increase in the pro-GN level in circulation; however, compared to the control group, the pro-GN concentration remained decreased. These results can be explained by the study conducted by Brenna et al. , where tIn group of patients with Crohn’s disease, the implemented anti-inflammatory treatment significantly decreased the concentration of PTX3, reaching levels similar to those measured in the serum of healthy individuals. As pro-inflammatory cytokines stimulate the synthesis and release of PTX3 from neutrophils and macrophages, the observed decline in PTX3 levels may reflect the diminishing of the pro-inflammatory state across the intestinal mucosa ,10. FurtIn our previous study, we assessed the serum profiles of pro-GN, PTX3 and S100A12 protein in patients with ulcerative colitis . In thisThe increase in PTX3 level in the serum of patients with ulcerative colitis compared to those with Crohn’s disease may be related to enhanced NET formation during UC, as noted in the study conducted by Dinallo et al. . The menAmong patients with Crohn’s disease and ulcerative colitis, another difference was noted in the case of S100A12, the concentration of which was increased in the serum of CD patients compared to UC. These results may be further confirmed by the Foell et al.’s  study, iRegarding the divergent phenotypes of Crohn’s disease and its possible extraintestinal manifestation, the diagnosis of the disease remains a significant challenge for clinicists. Therefore, the aim of our study was to identify new possible biomarkers of Crohn’s disease that could support the diagnostic process and disease monitoring. In our research, we assessed the concentration of a biomarker related to intestinal barrier integrity, proguanylin, and proteins related to inflammatory process, namely, pentraxin 3 and S100A12. The conducted statistical analyses revealed a significant difference in the serum profiles of pro-GN, PTX3 and S100A12 between patients with Crohn’s disease and healthy individuals, indicating the diagnostic usefulness of all the analyzed biomarkers in this disease. Moreover, a crucial relationship was noted between the level of PTX3 and the CDAI score, which assesses the disease activity, both before and after anti-inflammatory treatment. A similar correlation was observed for S100A12 serum concentration and disease activity but only in the post-treatment group of patients. These findings provide evidence that PTX3 and S100A12 measurements might be helpful in evaluating disease activity. Furthermore, we also estimated the effect of the implemented anti-inflammatory treatment based on prednisone on the profile of analyzed biomarkers and noted a significant difference in pro-GN and PTX3 levels, confirming the potential use of these parameters in evaluating the proper response to treatment. As in our previously conducted study, we assessed the levels of pro-GN, PTX3 and S100A12 in serum of patients with ulcerative colitis, and we also compared the profiles of these biomarkers among patients with CD and UC. We revealed a significant difference in the serum concentration of PTX3 and S100A12 protein between patients with CD and UC, highlighting the usefulness of these biomarkers in the differential diagnosis of IBD. However, the presented results should be interpreted with caution and confirmed on the greater study group. The results of our study not only provide new insights into the molecular basis of Chron’s disease but also can be helpful in the diagnosis and monitoring the disease’s activity.