Tenebrio molitor L., Acheta domesticus L., Alphitobius diaperinus P.) were used at two different additive levels (15% and 30%) in relation to the weight of the nuts. The addition of insect flour significantly increased protein content and the insoluble fraction of dietary fiber. The largest amount of these compounds was found in bars with 30% cricket flour, 15.51 g/100 g and 6.04 g/100 g, respectively, in comparison to standard bars, 10.78 g/100 g and 3.14 g/100 g, respectively. The greatest consumer acceptance was found in relation to bars with buffalo worm flour. The overall acceptance of these bars was 6.26–6.28 points compared to 6.48 for standard bars. Bars and raw materials were characterized by the high biological value of the protein. Cis linoleic acid dominated among unsaturated fatty acids. The percentage of this compound was in the range of 69.56%, for bars with a 30% addition of buffalo worm flour, to 73.88%, for bars with 15% cricket flour. Instrumental analysis of taste and smell compounds showed the presence of compounds such as 3-methylbutanoic acid, hexanal, and 2,3-pentanedione.Six types of nut-based bars with the addition of edible insect flour were obtained. Flours made from three different insects ( In recent years, the world has faced a series of new challenges, of which the issue of feeding humanity appears to be of the utmost importance. Global population growth is projected to reach 10 billion by 2050, posing enormous challenges to all sectors involved in food production. Classic animal production that provides humans with the main amount of protein containing the necessary amino acids may turn out to be insufficient. Breeding of cattle, pigs, and poultry requires a lot of energy, requires space, and is both water- and feed-intensive. To this should be added other environmental issues, such as greenhouse gas emissions . Edible Tenebrio molitor L.) and buffalo worm (Alphitobius diaperinus P.) at their larval stage, as well as house cricket (Acheta domestica L.). These insects are used to enrich, inter alia, products such as bread [Among the most frequently and willingly used edible insects in the creation of food products are yellow mealworm  of 2.7% between 2022 and 2030. The changing lifestyle seen mainly among Americans and Europeans, involving the consumption of smaller meals, contributes to the increased consumption of snacks. That is why bars, especially those which are a healthier form of snack product, are in growing demand. Changing consumer behavior, giving up traditional animal protein, is mainly due to people’s growing concerns about animal welfare or sustainable development. Consumers are increasingly looking for products that will help them maintain a healthy and balanced diet. Functional snacks are becoming more and more popular, and the expectations and needs of young customers are evolving at a rapid pace [T. molitor), buffalo worm (A. diaperinus), and house cricket (A. domesticus) powders.Nut bars are a type of food aimed at young, active people who want to quickly supplement the required food ingredients. These people are often more open to nutritional information and, at the same time, are highly aware of the ethical and environmental risks associated with food production. It is extremely important that, when facing the threat of failures in traditional food production, necessary alternatives to meet the nutritional needs of humanity are found. Promoting entomophagy among such people may contribute to increasing interest in eating edible insects in Western societies, or at least reducing the reluctance to introduce new food products to the market. This can be achieved by introducing products dedicated to young generations for whom they will soon become a natural alternative and nutritional choice. For this reason, it was decided that this study would focus on the development of nut-based bars enriched with powders from various edible insects. The most popular and consumer-accessible preparations of edible insects were used, i.e., yellow mealworm , fat (29.01), chitin (9.92), and insoluble fractions of dietary fiber (18.48 g/100 g) compared to the others . Nuts, e5.18, fatLactobacillus rhamnosus GG and the inhibition of Escherichia coli TG [Bifidobacteria and Lactobacillus.Chitin is a type of carbohydrate polymer that forms the exoskeleton of most arthropods, including insects. Selenius et al.  demonstr coli TG . Fernand coli TG  reportedOn the basis of electrophoretic profiles of raw materials, it can be concluded that the molecular weight of hazelnut proteins was in the range of about 8–70 kDa, with the main protein fractions approximately 17–20 kDa (two protein bands) and 32–37 kDa (three bands) . Cashew In the amino acid profile of both raw materials and the obtained bars, endogenous amino acids dominated; the highest sum of their content was characteristic for the standard bar, and the lowest for hazelnuts . Most bars with insect flour did not differ significantly in terms of the sum of endogenous amino acids . The staTherefore, the content of amino acids in the studied bars was compared to the model proposed by the FAO/WHO . Based oThe fatty acid profile of the bars was correlated with the fatty acid profile of the raw materials used for their production. Particular insect flours and nuts differed significantly in the content of individual fatty acids. Their profile was dominated by cis-oleic acids (C18:1(9) n-9) (from 82.44 for hazelnuts to 73.87% for bars with 30% addition of cricket flour) . The conThe obtained results, regarding fatty acid profile of insect flour, are consistent with the data presented by other authors ,15,17,25After assessing the acceptance of six samples against the control sample , it was On the basis of a detailed analysis, the most frequently declared features of smell, taste, and texture were selected. Odor analysis showed that popcorn, coffee, and almond were the most frequently identified odors among all samples . HoweverA nutty flavor was most pronounced in the case of bars with 15% and 30% buffalo worm content compared to the control sample . For theAn unpleasant taste was most often reported for the variant with 30% mealworm content, as demonstrated by odor identification.Among the control bar texture descriptors, the most frequently reported texture characteristics were the firm and cohesive texture expected for this type of food product. With the addition of 15% insect flour, the identified structure was hard and cohesive, but the addition of a larger amount of insect flour, regardless of the type, resulted in deterioration of the texture .Results of the analysis of aroma compounds in the control and edible insect flour-substituted bars analyzed by an electronic nose are presented as a heatmap . A totalThe results showed that the proportions of different insect flours contributed to notable differences in the concentrations of volatile compounds. In addition, it was observed that bars enriched with insect flours, regardless of the type of flour, had significantly higher 3-methylbutanal content than the control samples, which is responsible for pleasant caramel, fruity, and malty notes . The MaiTaste quality (distinguished between different tastes) and its intensity in water extracts made from the control and edible insect flour-substituted bars were evaluated using an electronic tongue with seven cross-selectivity sensors. The results showed that enrichment of bars with insect flour significantly affected the taste attributes of the studied samples A,B. The The second cluster only contained the bar with 30% cricket flour, which is characterized by the sweetest taste among the samples tested. The third cluster includes bars with mealworm flour. These samples are characterized as more bitter than other bars. The fourth cluster contained bars with buffalo worm flour, which are characterized by a pronounced sour taste. The observed relationship between increases in sourness, sweetness, bitterness, and saltiness and the fortification of bars with insect flour may be related to their high protein, mineral, and organic acid content ,31. TherT. molitor , buffalo worm A. diaperinus , or cricket A. domesticus  was added based on the weight of the nuts.Nut bars were made of hazelnuts  and cashew nuts , in a 1:1 ratio, and honeydew honey , in an amount equal to 0.75% of the weight of the nuts. In the enriched bars, 15% and 30% insect flour from mealworm Nuts were roasted in a pan at 180 °C for 5 min. Crushed nuts for standard bars and crushed nuts and insect flour for the remaining bars were combined with dissolved honey (35 °C), thoroughly mixed, placed in silicone molds with dimensions of 75 × 30 × 30 mm, and placed in a refrigerator at 4 °C for 120 min.Basic physicochemical analysis of standard bars, as well as bars with the addition of edible insect powder (flour), was conducted according to AOAC methods . Chitin Analysis was based on the methods of Moore and Stein , DavidsoTo express the nutritional value of the protein, amino acid score (AAS) was calculated according to the FAO/WHO (1991) (Equation (1)): the FAO .v/v glycerol, and DTT 50 mg/mL at 100 °C for 5 min. The Schägger-von Jagow method [Extraction was performed using a mixture containing 125 mM Tris-HCl pH 6.8, 4% SDS, 20% w method  with pro3 (methanol solution)) was added and heating was continued at 85 °C. To isolate the fatty acid methanol esters, 1 mL of hexane and 5 mL of saturated NaCl solution was added to the sample. Chromatographic separation was obtained using a Shimadzu GC2010Plus Chromatograph  with a flame ionization detector (FID). Operating parameters were as follows: FID detector temp., 250 °C; temperature dispenser, 220 °C; oven temp, 80 °C (held for 2.5 min), heated to 160 °C (60 °C/min) and held for 35 min; heated to 230 °C (15 °C/min) and held for 5 min; heated to 240 °C (5 °C/min) and held for 5.5 min. An SH-FAME column (30 m × 0.32 mm × 0.25 μm) was used and the carrier gas was helium (1.6 mL/min). Split ratio 100. Identification of individual fatty acids in the form of their methyl esters was carried out by comparing the retention times of the peaks of the standard substances  and the peaks of the analyzed samples. Analyses was performed in four repetitions.Fatty acid profile was determined based on the AOAC method . The extOrganoleptic analysis of the obtained bread was carried out by 36 panelists aged 20 to 29 who were selected randomly from students at the Medical University of Gdansk on the basis of a descriptive analysis test. During the planning stage of the experiment, the sample size was selected at a level generating statistical conclusions with appropriate accuracy and certainty, and the probability of the test detecting effects of a given size was examined based on test power analysis and interval estimation of the sample size. The performed statistical analysis showed that all the tests that were made for the acceptance assessment of the seven test samples of bars can be considered representative. Based on collected data from the assessment of acceptance level, the minimum sample size for each bar should be at least 24 for each variant for all measurements. The evaluators provided answers on the previously prepared evaluation cards. Each assessor received his own set of samples (six samples + control) in a random order. A seven-point Likert scale with appropriately selected descriptors was used for the evaluation: appearance, smell, texture, taste, and the overall acceptability of the bread were rated on a scale from one to seven . Additionally, the flavor, aroma, and texture profile of the bars obtained were determined. In the case of smell, the assessors had a choice of the eleven most frequently reported smells: eight positive smells  and three negative smells , with one identified as unpleasant. Regarding the taste assessment, nine basic tastes were identified: five desirable tastes  and four undesirable tastes , with one identified as unpleasant. Regarding the texture assessment, eight differentiators characterizing the texture and consistency of bars were identified: hard, crumbly, sticky, springy, adhesive, sandy, and grainy.Analysis of the three basic discriminants was based on the assessors selecting one answer out of five possible: one—completely imperceptible; two—almost imperceptible; three—hard to define; four—slightly perceptible; and five—completely perceptible.The evaluators were healthy, took no medications or supplements, and followed no special diets. All participants voluntarily signed consent forms in order to participate in the research, which was approved by the Independent Bioethical Committee for Scientific Research at the Medical University of Gdansk (NKBBN/346/2021). This study complies with the ethical principles of non-harm, charity, justice, and autonomy as enshrined in the ethical provisions of the revised 2013 Declaration of Helsinki. Before the study, all evaluators completed the Food Neophobia Study (FNS) questionnaire. Only those who did not show nutritional neophobia participated in the study.Electronic nose (E-nose) analysis of volatile flavor compounds was carried out according to the method of Żyżelewicz et al. , with sog for 10 min at 20 °C. The supernatants were filtered through Whatman no. 4 filter paper and were transferred to the glass containers of the 25 mL autosampler. Analysis time was set at 120 s at 25 °C and data from the last 30 s were used for acquisition. Between measurements, the sensors were rinsed with distilled water for 120 s. Prior to analysis, the e-tongue sensors were conditioned (0.01 mol/L HCl), calibrated (0.01 mol/L HCl), and tested  for proper function and stability. After successful testing, each water extract was analyzed five times. Instrument control, data acquisition, and data processing were performed with Alphasoft software . Taste screening analysis was used to rank the samples according to taste attributes on a 0 to 12 intensity scale. Principal component analysis (PCA) was performed to discriminate the response signals from the seven sensors. The heat map was created using a web-based interactive next-generation clustered heat map (NG-CHM) builder (https://build.ngchm.net/NGCHM-web-builder/) (accessed on 30 August 2022) with hierarchical clustering using the Euclidean distance metric with Ward’s agglomeration method.Instrumental taste analysis of samples was carried out using the Alpha MOS ASTREE II electronic tongue (e-tongue) instrument , which consists of a 48-position autosampler for automated analysis of a sample set in reproducible conditions , an array of liquid sensors (set #5), and a reference electrode (Ag/AgCl). The sensor set #5 array used in this study contains seven sensors  and was designed for food and beverage applications . For e-tp ≤ 0.05. When the Levene test indicated significant differences, a post-hoc Fisher’s least significant difference (LSD) test was performed. The results are presented as mean ± standard deviation. The primary data obtained in the consumer evaluation study were subjected to statistical calculations, and on their basis a regression analysis was performed for each of the tested insect meal variants. Multiple regression was used to quantify the relationships between multiple independent variables, such as appearance, smell, texture, and taste, and the dependent variable acceptance. Additionally, Dixon’s Q test was performed to eliminate gross errors. The value of Q was lower than the value determined from the Qcr tables each time, which means that the doubtful result was an element of the sample for α = 0.05.Statistical analysis was carried out using Statistica 13.0 . A two-way ANOVA for flour type, percentage of substitution, and their interaction was used to test bread features at a significance level of Analysis of the obtained products showed that organoleptic acceptance was influenced by the type of edible insect flour used. This was a result of both the taste and smell sensations, as well as the texture of the tested product. Bars with the addition of buffalo worm flour were rated the highest, which contrasts with bars with the addition of mealworm flour. This evaluation could be based on the volatile compound profile, in particular the presence of compounds such as 3-methylobutanoic acid, hexanal, and 2,3-pentanedione in mealworm flour. Bars enriched with buffalo worm and cricket flours were high in butanal and furfural, which give products a chocolate and caramel taste and which may have contributed to the higher organoleptic evaluation of these bars. Regardless of the above, all bars containing insect flour showed an improvement in nutritional characteristics, especially in terms of protein quality, which was characterized by high biological value; leucine and lysine were identified as the limiting amino acids.


Cast iron’s propensity for plastic deformation under complex loading conditions is analyzed from the standpoint of the morphology of graphite inclusions, depending on the degree and mechanical scheme of deformation. The research methodology included: the choice of a brand of cast iron with spherical graphite and the technological process of its deformation; mathematical modeling of the deformation process using the DEFORM–3D software package; stamping of an experimental batch of forgings; and microstructural studies of the forging material, together with the data of its stress–strain state and the direction of flow of the material. Under the conditions of comprehensive compression of the workpiece material at the beginning of the deformation process, lateral pressure was created from the tool walls. It was carried out by selecting the size and shape of the initial blank by mathematical modeling. When analyzing the morphology of graphite inclusions, the stress state scheme was determined as the main influencing factor. The greatest change in the shape and size of graphite inclusions of cast iron, including their crushing, corresponds to the condition of the disappearance of all-round compression. When stamping in open dies, this occurs in the area where the material exits from the stamp engraving into the flash gutter. In the process of forming, cast iron showed the possibility of deformation in the deformation intensity index ε As aIn mechanical engineering, one of the important practical tasks is the manufacture of products from hard-to-deform metal materials, one of which is cast iron. As a structural material, grey cast iron contains, in the microstructure, graphite, which is widely used for the manufacture of various parts. Compared with steels, it has a number of advantages: it works well on friction, is wear-resistant, and dampens vibration. However, cast iron is inferior in strength due to the low level of viscosity and fracture resistance, which is due, on the one hand, to the presence of graphite particles, and on the other, to defects in the structure of the cast metal ,2,3,4,5.0.5–0.6) . The sec0.5–0.6) ,11,12. TWhen analyzing the developed processes of plastic deformation of cast iron to date, the practical interest in the manufacture of volumetric shaped products using deformation technological schemes that differ from the schemes of the implemented processes should be noted. In particular, this applies to the process of hot volumetric stamping in open dies, which makes it possible to produce products of various shapes. The shaping of the material in open dies is characterized by uneven all-round compression at an average total pressure and incomplete lateral pressure of the metal on the rigid walls of the tool. During deformation, part of the metal is displaced from the engraving in the form of a technological burr, which causes local free broadening of the metal in the area of the exit into the burr groove, creating additional tensile stresses that contribute to destruction. The structural features of the deformation focus and the periods of its change during stamping in open dies are described in detail in the works of M.V. Storozhev, S.I. Gubkin, E. I. Semenov, and other scientists ,21. To dThe solution to this problem involves the application of a comprehensive research methodology, which includes: mathematical modeling of the process, with the determination of the optimal dimensions of the workpiece and the assessment of the stress–strain state and the nature of the flow of the material in its volume during deformation; stamping of the experimental batch of forgings; and structural studies of the deformed material.The strength characteristics are hardness 170–180 HB, which corresponds to the ultimate strength 490–530 MPa.h = 12%.The permissible degree of deformation in hot precipitation is εThe chemical composition of cast iron is shown in Ductile cast iron in a ferrite–perlite matrix with the following characteristics was selected as the material under study:h = 20%. It was also noted that the cooling of the deformed material must be carried out in the furnace at a speed of 100 °C per hour [In order to clarify the ductility parameters of cast iron used for deformation, tests were carried out on the hot sediment of turned cylindrical samples with a diameter of 30 mm and a height of 60 mm from the batch of material used. The samples were heated in the furnace to a temperature of 1025–1050 °C and deformed by precipitation on the press with degrees of deformation of 10, 15, and 20%. In order to avoid contact with cold strikers, the deflection was carried out between heated plates. The results obtained showed that the cast iron used for testing has a ductility with a degree of deformation at a hot draft of εper hour . As a teMathematical modeling was carried out using the DEFORM–3D software package .The optimal dimensions of the initial workpiece were determined based on the possibility of creating conditions of all-round uneven compression in it during deformation. According to the modeling results, the dimensions of the workpiece were selected: height 30 mm and diameter 56 mm, which is 4 mm less than the internal size of the flange section of the working cavity of the stamp.Evaluation of the stress–strain state in the volume of the product was carried out in areas corresponding to microstructural studies. The schemes of the deformed state of the workpiece material are shown in Stamping was carried out on a crank hot-stamping press with a force of 10 kN. The ground workpieces were heated in an electric furnace at a temperature of 1025–1050 °C for one hour. A sketch (a) and a photograph (b) of the stamped forging of the “flange” part are shown in Microstructural analysis was carried out on samples, the cutting scheme of which, from the fourth part of the flange’s forging, is shown in One feature of the shaping of the initial blank in an open die is the fact that individual sections have different directions of metal flow due to the implementation of various mechanical deformation schemes, which forms a multidirectional structure of graphite particles.The material of the initial blank is initially deformed in the central zone under conditions of reverse extrusion a. The stThe beginning of deformation of the material of the lateral  zone of the workpiece has a delay in relation to the deformation of the material of the central zone b. In itsIn the subsequent period of stamping, when the material flows out in the form of a technological burr, in the zone of the connector plane, the state of back pressure, as well as all-round compression, disappears. The deformed state of the material, as in the central zone, shortens in the direction of compression and elongates towards the center of the workpiece and the perimeter of the stamp. The maximum voltage of 239–256 MPa corresponds to the output of the material from the engraving along the connector plane.The formation of the cylindrical section of the product occurs last of all under conditions of reverse extrusion, which is carried out due to counter radial displacements of the material of the central and peripheral zones of the workpiece c. FrictiThe implementation of various mechanical deformation schemes during forging was expressed in the unevenness of the deformed state of the material in its volume and the change in shape and size of graphite inclusions. The location of the studied characteristic zones in the forging layers is shown in i = 2.02–2.52 and, in the second layer, it is εi = 1.64–1.9. The deformed state of the material displays shortening in the direction of compression and elongation in two other directions, primarily to the perimeter of the engraving stamp. Graphite inclusions have an elongated shape in the direction of the flow of the material into the burr groove (Along the perimeter of the flange part of the forging at the place where the material enters the burr groove, in the first layer, the deformation intensity has the value εr groove .i = 1.41–1.52 and then increases again to εi = 1.68–1.91. In the center, the deformed state of the material is characterized by shortening in the direction of compression and uniform elongation in the other two directions. Graphite inclusions have largely changed their spherical shape to a flattened lenticular shape, the position of which is perpendicular to the direction of precipitation (In the direction of the forging center, the deformed state of the first two layers decreases to the value of εpitation .i = 0.76–1.26, which is less than in the first two layers. As the transition to the pipe part of the forging occurs, the deformed state of these layers increases, reaching the value of εi = 1.79–2.21 in the radius section. The deformed state of the material is characterized by elongation of its height and shortening in thickness. The change in the shape of graphite inclusions is also characterized by stretching in the direction of extrusion (In the third and fourth layers of the perimeter of the flange part of the forging, the deformation has the value εxtrusion .i = 2.07–2.42. The highest value corresponds to the outer surface of the section, the smallest to the inner surface.In the fifth and sixth layers of the pipe section, the deformed state continues to increase to the value εi = 1.5, and in the inner layers, to the value of εi = 1.35. Graphite inclusions have slightly lost their rounded shape (In the seventh and eighth layers, the deformed state of the pipe section in comparison with the previous layers decreases: in the outer layers, to the value of εed shape .Based on the result of the deformation analysis, it can be concluded that there is a significant unevenness of deformation in the forging volume, which is determined by the complex geometry of the deformation focus and, as a consequence, by different directions of the material flow . The maii = 2.5.There were positive results of the stamping of the experimental batch of round forgings, in terms of the technological possibility of hot plastic deformation of ductile cast iron in open dies. In addition, the position that the deformability of the material is not a property, but the state of the material, was also confirmed and is determined by the conditions of deformation: speed, temperature, degree, and mechanical scheme of deformation. The most significant factor in increasing plasticity is considered to be deformation in a state of all-round compression. It can be created by applying special techniques that simulate hydrostatic pressure. In this work, it was created due to lateral pressure from the walls of the tool, carried out by optimizing the size of the initial workpiece. The diameter of the workpiece was selected in accordance with the dimensions of the inner contour of the flange part of the product. During deformation, the cast iron showed plasticity with a value of deformation intensity of εi = 2.1–2.5, in all zones, graphite inclusions had different morphologies, which showed their dependence on loading conditions. The most difficult loading corresponds to the zone of the material’s exit from the die cavity into the burr groove, in which there is no state of all-round compression. Unlike the other two zones, in which the scheme of uneven all-round compression is implemented, on the one hand, by a deforming tool, and on the other, by adjacent layers of metal, there is no compressive stress in the considered zone from the material outlet side. The morphology of graphite inclusions is characterized by the greatest change in shape and size, including their fragmentation.The study of structural changes in cast iron with spherical graphite during deformation in open dies made it possible to evaluate the ability of graphite inclusions to undergo plastic deformation under conditions of complex loading and large degrees of deformation. During forging, the material underwent the greatest deformation in three zones: along the perimeter and center of the flange part and in the transition zone of the flange part to the cylindrical section. With almost identical degrees of deformation, ε1.By the method of hot volumetric stamping in an open die, an experimental batch of forgings of the round shape of the “flange” part, with satisfactory quality of the deformed metal in macro and microstructure, grinding, and orientation of the graphite phase in the direction of the main deformations, was obtained.2.i = 2.5.The results of the described work made it possible to determine the technological possibility of stamping forgings of round and similar shape in open dies in terms of creating conditions for uneven all-round compression of the material, while cast iron showed plasticity with a value of deformation intensity ε3.When stamping in open dies to create conditions for all-round compression, it is rational to use special techniques that simulate hydrostatic pressure, in particular, lateral pressure from the walls of the tool, carried out by selecting the size and shape of the initial workpiece.4.The study of structural changes in ductile cast iron during deformation in open dies has shown the ability of graphite inclusions to undergo plastic deformation under conditions of complex loading with changes in shape and size, including their crushing.Based on the results of the work, the following conclusions were made:


In addition, the G/COFs/BMI composites showed lower friction coefficient fluctuations and volume wear rates than G/BMI composites. This is mainly attributed to the fact that the deposition of COFs can not only effectively prevent the aggregation of graphene nanosheets, but can also significantly improve the compatibility and interfacial bond between the graphene and BMI matrix. Moreover, the good synergistic effect between the lamellar COFs and graphene nanosheets can generate high-quality self-lubricating transfer films during the friction process. The excellent dispersibility, efficient chemical functionalization, better friction reduction and wear-resistance properties, and facile preparation method make graphene/COFs hybrid nanoparticles promising as an excellent lubricating filler.The poor compatibility between the polymer matrix and complex modification processes greatly affects the excellent tribological properties of graphene in the polymer matrix. In this study, a covalent organic framework (COF)-coated graphene hybrid lubricating filler (G/COFs) was synthesized in situ using a sample one-step mechanochemical synthesis process. This was used to improve the tribological properties of bismaleimide (BMI) resin. The morphology and microstructure of the G/COFs hybrid were characterized, and the effect of the added amount on the tribological properties of the G/COFs/BMI composites was studied. The results showed that the G/COFs hybrid could improve the stability of the friction coefficient and decrease the volume wear rate of BMI composites. Compared to the neat BMI, the 0.6 wt% G/COFs/BMI composites showed optimal tribological performance, with the friction coefficient and volume wear rate decreasing from 0.35 to 0.14 and from 48 × 10 Due to their significant mechanical strength and high-temperature resistance, polymer materials are widely applied in aerospace, army facilities, healthcare equipment, automatic processing, and other fields . Zhou etGraphene (G) has been widely introduced into the polymer matrix as a lubricant nanofiller to improve friction reduction and the wear-resistance of composites due to its high thermal conductivity, significant mechanical properties, and superior friction and abrasion properties ,8. HowevCovalent organic frameworks (COFs) ,19,20,21In the present work, with graphene oxide, p-Phenylenediamine, and 1,3,5-Benzenetricarbonyl trichloride as raw materials, a composite lubricating filler (G/COFs) was synthesized via a one-step mechanochemical synthesis. Following, the G/COFs hybrid was introduced into bismaleimide (BMI) resin and G/COFs/BMI composites were cast. The morphology and microstructure of the G/COFs were characterized by FT-IR, XRD, and TEM, and the effects of the added amount on the tribological properties of the BMI composite were studied. The results revealed that the tribological properties of the BMI composite were significantly improved by introducing the G/COFs hybrid. The use of COF-modified G not only avoids graphene nanosheet agglomeration, but the lamellar COFs and graphene nanosheets also have a good synergistic effect, resulting in the composites being able to generate high-quality self-lubricating transfer films during the friction process.2SO4, 98%) and concentrated hydrochloric acid  were obtained from Sinopharm Chemical Reagent Co., Ltd. . Potassium persulfate , phosphorus pentoxide , and anhydrous ethanol (99.7%) were sourced from Tianjin Fuyu Fine Chemical Co . Natural graphite flakes (325 mesh) were obtained from Qingdao Hensen Graphite Co., Ltd. . Sodium hydroxide , p-Phenylenediamine (97%), and 1,3,5-Benzenetricarbonyl trichloride (98%) were obtained from Shanghai Macklin Biochemical Co. . 4,4′-Bismaleimidodiphenylmethane  and diallyl bisphenol A  were purchased from Wuhan Zhisheng Technology Co. . All reagents were applied without further purification.Concentrated sulfuric acid (H2S2O8 (5 g) and P2O5 (5 g) were mixed and dissolved in H2SO4; then, the graphite was added to an oil bath at 80 °C and stirred and kept for 6 h. After cooling to room temperature, the mixture was rinsed with deionized water to neutralize it, and the powder was dried at 60 °C for 12 h to obtain a black powder. KFirstly, graphene oxide (100 mg), p-Phenylenediamine (140 mg), and 1,3,5-Benzenetricarbonyl trichloride (160 mg) were added to a grinding pot with a certain number of agate balls and ground in a low-temperature planetary ball mill at 500 r/min for 90 min. Next, the pot was filled with 80 mL of 5% aqueous sodium hydroxide solution, and milling was maintained for 30 min. Finally, the product was filtered through a filter membrane and washed with deionized solution and ethanol, and it was finally dried under vacuum at 70 °C for 12 h. The black powder obtained was the prepared G/COFs.The G/COFs/BMI composites were prepared by mixing pre-weighed quantities of G/COFs, DBA, and BDM (DBA and BDM with a mass ratio of 3:4). Then, the mixture was heated to 135 °C until the mixture had totally melted and the G/COFs had uniformly dispersed. Finally, the mixture was cured following the schedule of 150 °C/2 h + 180 °C/2 h + 220 °C/4 h. The post-curing process was 250 °C/4 h. The composites based on BMI, including neat BMI and G/BMI composites with different filler contents ,34, were2 atmosphere at a heating rate of 10 °C/min. The Fourier transform infrared  spectra of samples were characterized. The sample phases were determined by an X-ray diffractometer . The sample morphology was recorded by a transmission electron microscope . A small amount of the sample was dispersed into ethanol and sonicated to make it uniformly dispersed. The sample was dropped onto a 230-mesh carbon support film to obtain a transmission electron microscope sample. The surface morphology of the samples was observed using a scanning electron microscope . The tribological properties of the samples were examined using a tribological test machine . The experiments were conducted with end-face friction sub-materials made of 45# steel, and friction and abrasion tests were performed on a testing machine under dry conditions. The hardness properties of the composite materials were tested by a microhardness tester  Co., Ltd., Suzhou, China). The thermal properties of the composites were analyzed using a thermogravimetric analyzer  in a N−1, which belong to the C=O stretching vibrations and C−Cl stretching vibrations in the acyl chloride groups, respectively [−1 belong to the anti-symmetric and symmetric stretching vibration peaks of −NH2, respectively. In the spectrum of the COFs, the peaks at 3051 cm−1 and 1514 cm−1 belong to the C−H and C=C stretching vibrations of the benzene ring, respectively. The peaks at 824 cm−1 and 708 cm−1 belong to the characteristic substitution peaks on the benzene rings [2 stretching band (3202 and 3380 cm−1) in p-Phenylenediamine and the C−Cl stretching (704 cm−1) in 1,3,5-Benzenetricarbonyl trichloride. Meanwhile, two new peaks occurred at 1253 cm−1 and 1660 cm−1, which were assigned to the absorption III (C=N) and Ⅰ (C=O) bands of −CONH− [−1, which belong to the C−O−C stretching vibrations, C−H bending vibration, and −OH stretching vibrations, respectively [FT-IR was used to study the chemical structures of p-Phenylenediamine, 1,3,5-Benzenetricarbonyl trichloride, graphene oxide, COFs, and G/COFs. As shown in ectively . In the ne rings ,28. In af −CONH− ,36. Thesectively . The FT-XRD analysis was used to study the structure of the sample. To identify the successful fabrication of COFs and G/COFs, the morphological structure of the samples was analyzed by TEM. As shown in −6 mm3/(N·m). Compared to neat BMI (0.35 and 48 × 10−6 mm3/(N·m)), they decreased by 60% and 77.9%, respectively. The reduction of the friction coefficient of the G/COFs compared to BMI is significantly better than similar reductions seen in the literature [The friction coefficient and volume wear rate of composites with various filler contents are shown in terature ,45. Howeterature .To obtain the tribological mechanism of G/COFs, the tribological properties of neat BMI, 0.6 wt% G/BMI composites, and 0.6 wt% G/COF/BMI composites were studied, as shown in To further investigate the tribological suitability of G/COFs/BMI composites, the tribological properties of G/COFs/BMI composites under different loads and rotational speeds were studied at 0.6 wt% filling. To study the wear mechanism of G/COFs/BMI composites, SEM was adopted to analyze the worn surface of the composites. Because of the enormous amount of heat generated during the friction process, the thermal stability of the composites is critical for their tribological qualities. Hardness is a parameter that resists various permanent shape changes when subjected to forces, and the hardness of a material is strongly correlated with its frictional properties. The correlation between the hardness of G/COFs/BMI composites and the G/COFs content is shown in −6 mm3/(N‧m). These are lowered by 60% and 77.9% compared to neat BMI, respectively. Even in harsh friction environments, the tribological characteristics of the G/COFs/BMI composites are still superior to those of neat BMI. This is mainly attributed to the fact that COF-coated graphene can not only effectively improve compatibility with the polymer matrix, but can also effectively play a role in friction reduction and wear resistance, thus synergistically improving the tribological properties of their composites. We believe that these novel G/COFs hybrid nanoparticles have the potential to become the lubricating filler for large-scale production applications, such as for the production of various automotive parts. However, this method is limited to G/COFs hybrid nanoparticles that need to be synthesized at a high temperature. In summary, novel G/COFs hybrid nanoparticles were synthesized in one step via mechanochemical synthesis. This method not only avoids the use of harmful solvents, but can also realize the scaled-up production of composite nanomaterials. The G/COFs/BMI composites show superior tribological characteristics compared to neat BMI and G /BMI composites. When the G/COFs addition is merely 0.6 wt%, the frictional coefficient and volume wear rate of their BMI composites are decreased to 0.14 and 10.6 × 10


Cyclin-dependent kinase-5 (CDK5) is a key kinase involved in brain development and function and recently found to be involved in neuronal and astroglial apoptosis, neural stem/progenitor cell stemness, mitochondrial fission, and synaptic transmission. But the specific mechanism of CDK5-mediated anti-inflammatory remains unclear in ICH. The aim of the present study was to explore the role of CDK5 in mediating microglia activity through activated DRP1 phosphorylation in a rat ICH model. α, interleukin- (IL-) 1β, and IL-6 levels were measured using enzyme-linked immunosorbent assay (ELISA).  We measured behavioral change after ICH; detected the expression of CDK5 in the rat brain using immunohistochemistry; and measured the protein levels of CDK5, p35, p25, p-histone H1, and p-DRP1 using Western blot analysis. Coimmunoprecipitation analysis indicated interaction of CDK5 and DRP1. Tumor necrosis factor-p < 0.05), and the expression of p35 was lowest at 12 h, while the expression of p25 peaked at 2 d after ICH. Besides, p-DRP1 expression change follows with CDK5 kinase activity change. Coimmunoprecipitation showed that interaction between CDK5 and DRP1 certainly exists in microglia. Then, knockdown CDK5 or p35 expression by siRNA reduced the expression level of p-DRP1. ELISA data showed that the protein levels of proinflammatory mediators, such as TNF-α, IL-1β, and IL-6, were decreased by knockdown of CDK5.  After ICH, CDK5 protein level and kinase activity increased. Western blot data showed that CDK5 expression increased from 6 h and peaked at 2 d after ICH ( CDK5 may regulate DRP1 by direct phosphorylation in microglia and further induce microglia secreting proinflammation factor. Intracerebral hemorrhage (ICH) is a subtype of stroke with high mortality and morbidity. The prevalence of ICH in the elderly is increasing rapidly, and this disease seriously endangers the health and life of elderly patients . When thMicroglia are extremely important for hematoma and debris clearance, but hyperactivation is also a factor in secondary inflammation in ICH . MicroglCDK5 is a unique member of cyclin-dependent kinase (CDK) family and is activated by p35 and p39 . During β-amyloid by microglia by upregulating the expression of lipoprotein lipase -ATP , and 125 μg/ml histone H1 was added to the protein samples, followed by incubation at 30°C for 30 min. The kinase reaction was then terminated by addition of 4× SDS loading buffer. Samples were boiled for 5 min and separated by 15% SDS-PAGE, and phosphorylated histone H1 was detected by immunoblotting with anti-phospho-histone H1-specific antibody .BV2 cells were treated with 30 μM). Twenty-four hours after stimulation, culture supernatants were collected and centrifuged. Concentrations of TNF-α, IL-1β, and IL-6 were measured with commercially available ELISA kits , followed by measurements in a plate reader at a wavelength of 450 nm.Cultured BV2 microglia were treated with hemin  .Western blot was performed to detect the time change of CDK5 expression near the hematoma. It was found that the expression of CDK5 in the sham operation group was low, and it began to increase at 6 h after ICH and reached a peak at day 2 after ICH ( Figures . We also Figures . Our resTo explore if CDK5 kinase activity changes in the rat ICH model, phosphorylation of histone H1 by activated CDK5-associated kinase activity was carried out. We found that p-histone H1 level upregulated at day 1 and reaching peak at day 3, then decreased following ICH Figures . DRP1 isSince most of CDK5 functions are concerned with its phosphorylation ability, and CDK5 interacts with phosphorylate DRP1 has been reported in neuron. Next, we use coimmunoprecipitation to define if CDK5 interacts with phosphorite DRP1 in microglia. The result of coimmunoprecipitation shows that interaction between CDK5 and DRP1 certainly exists in microglia . To defiα, IL-1β, and IL-6 by ELISA. The result shows that the expression of proinflammatory cytokines is induced by hemin stimulation in microglial cells. And the increment is blocked by knockdown of CDK5 (Figures In the past research, mitochondrial fusion and fission are mentioned in maintaining mitochondrial functions including mitochondrial biogenesis and distribution as well as apoptosis, cell division, and complementation of mitochondrial DNA . Park et Figures . It indiICH is a devastating cerebrovascular disease that includes primary injury and secondary injury . Inflammβ phagocytosis by microglia and provide a new insight into the molecular pathogenesis of AD [In recent years, abnormal CDK5 expression is associated with some neurodegenerative or cardiovascular diseases. Specific CDK5 function in microglia was first explored . They fois of AD . Microglis of AD . To furtis of AD . DRP1 esis of AD . CDK5 anis of AD . Since CHowever, our study has its own limitations. Due to the simplicity of the experimental design, some experimental data cannot fully enrich the accuracy of the experimental results. The specific regulatory mechanism between CDK5 and DRP1 has not been deeply explored. Therefore, in the next study, we will further explore the relationship between CDK5 and DRP and the regulatory mechanism, so as to provide an important basis for the pathology of ICH diseases.In a conclusion, our result shows DRP1 as a phosphorylated substrate of CDK5 in microglia, and the phosphorylation of DRP1 promotes the secretion of proinflammatory mediators by microglia in the ICH model. It may provide a therapeutic target of ICH.


In the process of foundation pit excavation, different retaining structures and support stiffness may lead to different deformation modes of retaining structures. The soil displacement cause by the deformation of the retaining structure is calculated by the virtual image technique. Meanwhile, the collaborative deformation model for rotation and dislocation is introduced to analyze the tunnel longitudinal deformation caused by different deformation modes as well as the tunnel maximum displacement equivalent field, and two case verifications are carried out. The study shows that there is a large difference in the size and distribution law of the soil displacement field caused by different enclosure structure deformation modes. The horizontal displacement of soil caused by the cantilever type always shows a "cantilever type" curve with increasing horizontal distance from the enclosure structure, while the composite type, inner convex type and kicker type develop from the "bow" to the "cantilever type" curve. The vertical displacement field of the soil is in the shape of a "spoon" and the soil exhibits a certain bulge deformation below the critical depth. The critical depths of the composite type and inner convex type are similar, while the kick type is the largest and the cantilever type is the smallest. The influence area of the maximum horizontal displacement of the tunnel outside the pit of the inner convex type and the composite type is basically the same. The influence area of the cantilever mode is the smallest, while the influence depth of the kick-in mode is higher than that of the other three deformation modes. The closest distance was approximately 33 m between a construction foundation pit and Nanning Metro Line 22. The unloading effect on the sidewall due to the excavation of the foundation pit leads to the deformation of the enclosure structure, which generates additional loads on the adjacent tunnel. The excessive loads will endanger the safety of the tunnel. An engineering case in which the tunnel section of the Panchiao line was damaged due to adjacent foundation pit excavation during the construction of the Taipei MRT system was reported3. As a result, it is important to predict the deformation of adjacent shield tunnels due to foundation pit excavation.With the continuing development of rail transit, excavation projects of foundations adjacent to shield tunnels are becoming more common. For example, the minimum horizontal distance between the metro and the foundation pit was only 18.1 m6, numerical simulation7, centrifuge testing8 and measured data analysis10. In theoretical calculation, the two-stage analysis method is the most widely used method: first, calculating the soil unloading stress or displacement caused by the foundation excavation, then, computing the deformation of the tunnel subjected additional stress or displacement. For the unloading stress on the sidewall of the pit caused by the excavation of the pit, most of the current studies consider the active earth pressure outside the pit as the sidewall unloading amount12, the support function of the enclosure cannot be considered. Some literatures14 use the unloading loss rate β to consider the partial release of unloading stresses in the sidewalls. However, during the excavation of the foundation pit, the enclosure is bound to deform to varying degrees, thus affecting the amount of unloaded load on the enclosure. Zhang et al.15 derived the formula for calculating the unloading capacity considering the displacement under the convexity deformation pattern. It has been shown that there are differences in unloading on the sidewalls for different modes of envelope deformation, thus having different effects on the shield tunnel outside the pit16. However, studies on sidewall unloading under other deformation modes of the envelope have not been reported.This kind of engineering problem has received attention at home and abroad. At present, the main research methods are theoretical calculation5. However, the Euler–Bernoulli beam cannot take into account the shear effect between the rings, and thus it leads to a significant overestimation of the shear effect. The Timoshenko beam, which takes into account both bending and shear effects, are also commonly used to simulate shield tunnels13. However, the Timoshenko beam is still unable to simulate the assembling characteristics of segment rings. To fully understand the tunnel deformation mechanism, a few studies17 built a collaborative deformation mode with rotation and dislocation to simulate the shield tunnel.In the presented analytical methods, the shield tunnels are mainly considered as Euler Bernoulli beams as a way to simulate the bending deformation of the tunnelThis paper summarizes the formulas for calculating the deformation curve of pit sidewalls under four typical enclosure deformation modes. Meanwhile, the soil displacement fields caused by different enclosure deformation modes are calculated based on the virtual image technique, and the horizontal action range and influence depth of different enclosure deformation modes are compared and analyzed. In addition, this paper further introduces a rotating staggered platform collaborative deformation tunnel model that can fully take into account the tube assembly characteristics and investigates the difference in longitudinal deformation of the side-by-side shield tunnel under different pit envelope deformation modes.18. The four deformation modes are shown schematically in Fig. The deformation mode of a foundation pit enclosure structure is influenced by the form of foundation support and construction methods. There are four main deformation modes: cantilever, kick-in, convex and composite19.For flexible enclosure structures, when no support is set or excavation is shallow and no support has yet been set, there is a cantilevered distribution with maximum top displacementB, the excavation depth is d, and the depth of the enclosure structure is H. The enclosure structure underwent cantilever deformation. The deformation of enclosure structure at depth η is u(η). Gu et al.20 simplified the deformation of the row pile support to the cantilever deformation mode and provided the deformation curve of the row pile. Several scholars23 used the same hypothetical curve as the cantilever deformation of the enclosure structure.u denotes the displacement of the sidewall to the pit at depth z from corner y of the pit, H indicates the length of the enclosure structure, L corresponds to the length of the pit, and As indicated in Fig. y = L/2, substituting into Eq.  indicates the incremental deformation of the pit sidewall enclosure at depth z caused by the excavation of the ith layer, and di denotes the depth of the excavation surface after the excavation of the ith layer. Moreover, δimax  denotes the maximum incremental deformation of the pit sidewall enclosure caused by the excavation of the ith layer.As indicated in Fig. 15 used the ratio of the accumulated maximum deformation of the enclosure to the excavation depth δmax/d as the control parameter of the enclosure deformation, and they assumed that the accumulated deformation of each layer of excavation satisfies the control value. Accordingly, the maximum deformation increment of the excavation of the ith layer can be expressed as follows:Zhang et al.n layers and the depth of the excavated surface reaches di, the cumulative deformation distribution of the enclosure structure can be expressed as follows:When the foundation pit is excavated in 25 provided a formula for the lateral deformation curve of the enclosure wall on the symmetrical surface of the pit. Zhang et al.26 and Zhu et al.27 also used the formula to predict the composite deformation of the enclosure structure, which is as follows:Figure 28 proposed the virtual image technique to derive a formula to mathematically describe the distribution law of the surrounding soil displacement field owing to stratigraphic losses at any point in the elastic half-space. In this study, that method was used to calculate the soil displacement caused by the deformation of the pit sidewalls. The displacement mechanism of the soil loss involves certain simplifications with the following assumptions: (a) The soil is incompressible, and the soil displacement field is only due to soil loss. (b) The calculation process does not consider pore water pressure, soil consolidation, etc. (c) The same size of deformation occurs in each section of the envelope structure.In 1987, Sagastetax-axis component of the gap of radius a at the point F and the point P.Taking the convex deformation model as an example, Fig. x-axis component is as follows:F′ and the point P.Equation  represenDuring the establishment of Eq. , a shearG denotes the soil shear modulus.The additional shear stress generated at the ground surface is expressed as follows:x-axis generated by the shear stress at the point P is expressed as follows:When additional shear stress is applied to the surface with the opposite sign, the displacement component along the x-axis generated by a void of radius a at the point F in a semi-infinite space at the point P is as follows:In summary, the component of the soil displacement along the v generated in the elastic half-space space, as shown in Fig. n differential segments, each of which is approximated as a rectangle. The area corresponding to each differential segment is equated to a circle based on the area equivalence principle, and the equivalence radius was Since the outside of the enclosure should be 1/2 half-space, based on the symmetry, the problem can be transformed into a solution of deformation at any point under the stratigraphic loss of 2Similarly, the vertical displacement of the soil caused by the section deformation of the envelope structure is30 all control the maximum deformation of the enclosure structure. However, even if the maximum deformation of the enclosure structure is the same, the displacement field of the soil outside the pit will be different due to the different deformation modes of the enclosure structure. Therefore, it is of great significance for practical engineering to analyze the difference in soil displacement caused by different enclosure structures under the same maximum deformation of the foundation pit enclosure.During the excavation of the foundation pit, the current relevant specificationsL = 68 m, B = 72 m, excavation depth d = 15.8 m, underground diaphragm wall is 37.2 m below the ground, and the minimum distance between the sideline of the foundation pit enclosure and the tunnel axis is s = 12.6 m31. Statistics of the measured data show34 that the excavation depth of the foundation pit is closely related to the accumulated maximum deformation of the foundation pit enclosure. The δmax/d of the foundation pit enclosure in Hangzhou varies from 0.09 to 0.61%32. The deformation of foundation pits in Shanghai calculated by Xu et al.33 is very close to that collected by Qu et al.34 in Taipei, China, with δmax/d ranging between 0.1 and 1.0%, with an average value of 0.42%.Take the deep foundation pit project next to the shield tunnel of Metro Line 2 that has been put into operation at the intersection of Shixin Road and Jincheng Road in Xiaoshan District, Hangzhou City as a case. The excavation size of the foundation pit on the side of the tunnel is δmax/d of this project case are taken as 0.6%. Under the condition of the same cumulative maximum deformation, the deformation curves of the sidewall under the four deformation modes of the foundation pit enclosure are shown in Fig. The four envelope deformation modes Figure As shown in Fig. Figure The horizontal soil displacement field generated by the cantilever deformation mode of the envelope structure is shown in Fig. Figure As shown in Fig. The soil settlement outside the pit presents a "spoon"-shaped distribution. Figure The horizontal displacement field of soil caused by the cantilever deformation mode of the envelope structure is shown in Fig. Under the four deformation modes of the foundation pit enclosure, the soil will have a certain degree of uplift below a certain depth. The critical depth is related to the deformation mode of the envelope structure. The critical depth of the composite type is similar to that of the convex mode, the kick-in mode is the largest, and the cantilever mode is the smallest.15.This paper cites a cooperative deformation model that can comprehensively consider the two deformation effects of segment ring rotation and dislocation of shield tunnelsAs shown in Fig. l is the calculated longitudinal position of the tunnel, w(l) is the horizontal displacement distribution of the shield tunnel beside the foundation pit, and wt(l) is the horizontal displacement of the soil around the tunnel.Assuming that the shield tunnel beside the foundation pit and the surrounding soil meet the deformation coordination conditions, the displacement of the tunnel is equal to the displacement value of the soil at the corresponding position:m and m + 1 are the serial numbers of the two adjacent segment rings, and Dt is the ring width of the segment rings.The horizontal displacement between rings can be expressed asN is the number of segment rings within the calculation range of the shield tunnel. In theory, the larger the value of N, the higher the calculation accuracy. However, the corresponding calculation amount will also increase, and the calculation efficiency will be affected.(1) Work done by horizontal additional loads caused by soil deformation:Overcome the resistance of the formation to do work:Work against interring shear force:Overcome the tension caused by the rotation angle between the rings to do work:According to the process analysis of the longitudinal deformation of the shield tunnel, the total potential energy of the shield tunnel deformation can be specifically composed of the following four parts:Therefore, the following can be obtained:The total potential energy of deformation of the shield tunnel beside the foundation pit isksl and kt are the interring shear stiffness and interring tensile stiffness of the tunnel, respectively35. k is the subgrade coefficient of the soil, calculated by the Vesic36 formula, D is the outer diameter of the tunnel; μ is Poisson’s ratio of the soil; Es is the compressive modulus of the foundation soil; and (EI)eq is the equivalent flexural stiffness of the tunnel37.In Eqs. –22), ks, ks22), n is the Fourier expansion series.The distribution of the excavation area in the simplified model in this paper is parallel to the side shield tunnel. In theory, the longitudinal deformation of the shield tunnel should be symmetrical about the middle point of the excavation of the foundation pit, so it can be obtained by Fourier series expansion according to the cosine function:Ep takes the extreme value of each undetermined coefficient,ai is the (i + 1)th element in matrix A, i.e., the tunnel deformation curve function coefficient polynomial.Based on the principle of minimum potential energy, the total potential energy The governing equation can be obtained by solving the above equation:AT can be obtained:Calculated by Eq. , the undAT back into Eq.  are are29) aL = 68 m, B = 72 m, excavation depth d = 15.8 m, and the underground diaphragm wall is 37.2 m below the ground. The minimum distance between the sideline of the foundation pit envelope and the tunnel axis is s = 12.6 m31. The outer diameter of the shield tunnel lining is D = 6.2 m. Using C50 concrete segments, thickness t = 0.35 m, and ring width Dt = 1.2 m. The segment rings are connected by sixteen M30 longitudinal bolts. According to the calculation, ksl = 2.23 × 106 kN/m, kt = 9.39 × 105 kN/m, and (EI)eq = 1.1 × 108 kN m2. The tunnel axis is buried at a depth of 14.3 m. The soil in the project site is composed of silty clay, sandy silt, silty sand mixed with silt, and muddy silty clay from top to bottom. The compression modulus of each soil layer is 5.2 MPa, 10 MPa, 12.6 MPa and 3 MPa. The soil layers within the excavation area of the foundation pit from top to bottom are silty clay, sandy silt and silty sand mixed with silt, respectively. The tunnel is predominantly passing through the sandy silt, silty sand mixed with silt and muddy silty clay. According to the actual engineering geological conditions, Poisson's ratio of the soil is μ = 0.4 and Es = 6.39 MPa. The measured data of tunnel deformation is from the literature31.A foundation pit is located next to the shield tunnel of Hangzhou Metro Line 2, where δmax/d = 0.6%, N = 300, and the proportional coefficient of the rotation effect of the shield tunnel jx = 0.2. Figure In the calculation example, As shown in Fig. −5 rad.As shown in Fig. Figure 38. Suzhou Line 1 is located on the south side of the pit, parallel to the pit, with a distance of about 10.5 m from the enclosure structure, and the tunnel axis burial depth is about 13 m; calculation parameters ksl = 2.23 × 106 kN/m, and kt = 9.39 × 105 kN/m. Within the excavation area of the foundation pit, there are mainly powdered clay mixed with clay and silty. The tunnel interval is located in powdered clay with soil Poisson's ratio μ = 0.35 and Es = 10.76 MPa. The measured data of tunnel deformation is from the literature38.The plan size of the new office building pit of Hengfeng Bank is 85 m × 45 m, excavation depth is 10.6 mδmax/d = 0.5%, N = 100, and the rotational effect proportional coefficient of the shield tunnel beside the foundation pit is jx = 0.2. Figure In the calculation example, As shown in Fig. The horizontal displacement distributions of the shield tunnel caused by the convex type and the composite type are similar, and the horizontal displacement of the tunnel under the two deformation modes is in good agreement with the measured value. Similarly, the kick-in and cantilever deformation modes underestimate the horizontal displacement of the tunnel to a certain extent. In case 2, the influence range of the tunnel deformation caused by the four envelope structure deformation modes is also approximately twice the length of the sidewall of the foundation pit along the tunnel axis.With the development of pit design technology, the support scheme of the adjacent existing shield tunnel gradually develops into a strong support scheme with a large stiffness enclosure structure and multiple internal supports, and the deformation mode of the enclosure structure is mainly composite as well as convex type. In this paper, the construction time of case 1 is 2014 and the construction time of case 2 is 2017, so composite, as well as convex calculation results in the two cases, are more consistent with the actual measurement results.39. In this section, the horizontal displacement control value of the tunnel is 10 mm. In the composite envelope deformation mode, the tunnel horizontal displacement of more than 10 mm is approximately 46 m from the horizontal distance of the envelope structure, the depth is 30 m within the triangular area, and the range is similar to the convex envelope deformation mode. Both deformation modes reach the maximum horizontal displacement of the tunnel in the area near the pit bottom. In the composite and internal convex deformation modes, the maximum horizontal displacement of the tunnel decreases with increasing horizontal distance and depth from the enclosure, and the effect of depth is significantly greater than the effect of horizontal distance from the enclosure.Taking engineering case 1 as the basic working condition, Fig. Figure Figure The horizontal displacement fields of soil caused by the deformation modes of composite and convex enclosures are similar in distribution. Except for the horizontal displacement of the soil outside the pit caused by the cantilever-type deformation mode, which always shows a "cantilever-type" curve, the horizontal displacement of the deep soil outside the pit caused by other deformation modes develops from the "bow" to the "cantilever-type" curve with increasing horizontal distance from the enclosure structure.The vertical displacement field of the soil outside the pit caused by the deformation mode of four enclosure structures shows a "spoon" shape. The horizontal influence range of soil settlement caused by the four modes is, in descending order, kick-in, composite, convex, and cantilever type. After reaching critical depth, the soil will show a certain uplift deformation. The critical depths of the composite type and convex type are similar, while the kick-in type is the largest and the cantilever type is the smallest.For the four deformation modes on the horizontal displacement impact area of the tunnel outside the pit, the impact area of the convex type and the composite type is basically the same. However, since there is a certain displacement at the top of the composite enclosure, the range in the horizontal direction is slightly larger than that of the convex type. The cantilever mode has the smallest influence area, while the kick-in mode has the largest deformation at the bottom of the envelope, and the influence area is smaller in the horizontal direction but increases in the depth direction compared with the other three deformation modes.There is a large difference in the deformation of the side shield tunnel caused by different envelope deformation modes for the same cumulative maximum deformation of the envelope structure. During the construction of the foundation pit, the maximum displacement of the enclosure structure does not completely reflect the impact of the enclosure structure deformation on the tunnel outside the pit. In the predesign stage of the pit, it should be combined with the relative position of the pit and the tunnel to design a suitable support scheme, control the deformation mode of the enclosure structure and minimize the impact on the surrounding environment.Considering the deformation mode of the retaining structure, this paper proposes a simplified calculation method of the horizontal displacement of the shield tunnel caused by the excavation of the foundation pit, which can accurately evaluate the influence of the excavation of the foundation pit on the deformation of the tunnel. From the case conditions of this paper, the following four main conclusions are drawn.IN this research, the influence of different deformation modes of foundation pit retaining structures on the deformation of the surrounding soil is analyzed. At the same time, a simplified calculation method for the deformation of the side shield tunnel caused by the excavation of the foundation pit is proposed, which can calculate the rotation and dislocation of the shield tunnel during the deformation process.15 on the calculation method of shield tunnel deformation beside the foundation pit used the stress release amount of the side wall of the foundation pit to evaluate the tunnel deformation. However, the determination of the stress release amount is difficult and cannot be verified. The method in this paper evaluates the deformation of the side tunnel by the deformation of the side wall, which can be determined by the measured engineering data or the specified deformation control value and can be easily verified.The previous researchAs the calculation results in this paper show Figs. , 15, 16,14 simplified the shield tunnel to the Euler–Bernoulli beam or Timoshenko beam, but could not consider the characteristics of shield tunnel segment assembly. In this paper, the rotating staggered platform model is introduced to simulate the rotation and staggered platform behavior of the shield tunnel segment during deformation.Most previous studiesCompared with previous studies, the calculation method in this research has the following advantages:Many simplifications are made in the analysis process, such as ignoring the influence of tunnel existence, precipitation, stratification of foundation soil and nonlinear action of tunnel and soil. No consideration is given to the nonuniformity of soil convergence, and the theoretical results will have certain errors, which can be further studied based on this study.δmax/d plays an important role in the calculation results. There are three methods to take the value of parameter δmax/d, namely, the measured data, the control value specified in the specification or design, and the parameter back analysis. To accurately evaluate the deformation of the side shield tunnel caused by the excavation of the foundation pit, the accurate value of parameter δmax/d is necessary. However, due to the regional differences in the soil mass and the different supporting conditions of the foundation pit, the parameter δmax/d of different projects varies greatly. The subsequent statistical analysis of the parameter δmax/d of different soil mass conditions and different regions will be carried out to help the accurate value.The parameter However, the calculation method in this paper still has some shortcomings:


But estimation of domain structure from its analysis is difficult due to the wide overlap of spectra from multiple domains. Therefore, we have developed a materials informatics approach that combines the domain modeling (http://dmar.riken.jp/matrigica/) and the integrated analysis of meta-information  in polymer materials. Firstly, the 1H-static ssNMR data of 120 polymer materials were subjected to a short-time Fourier transform to obtain frequency, intensity, and T2 relaxation time for domains with different mobility. The average T2 relaxation time of each domain is 0.96 ms for Mobile, 0.55 ms for Intermediate (Mobile), 0.32 ms for Intermediate (Rigid), and 0.11 ms for Rigid. Secondly, the estimated domain proportions were integrated with meta-information such as elements, functional group and thermophysical properties and was analyzed using a self-organization map and market basket analysis. This proposed method can contribute to explore structure–property relationships of polymer materials with multiple domains.In the development of polymer materials, it is an important issue to explore the complex relationships between domain structure and physical properties. In the domain structure analysis of polymer materials,  Computational science solves equations numerically based on theory and physical models. On the other hand, data science explores candidate materials with a certain function from a large quantity of material data. Its physical meaning needs to be verified by experimental, theoretical and computational science. The meta-information involved in the production process of the material also plays an important role in desired materials development10.Traditional design approaches for materials are experimentally driven, facing significant challenges due to the vast design space of materials. Experimental science can be supported by materials informatics12. Furthermore, “carbon–neutral” bio-based polymers, such as polylactic acid (PLA), polybutylene succinate (PBS), poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBH), and poly-ε-caprolactone (PCL), have become a focus in the era of biorefinery materials as an alternative to oil-based materials14. Polymers such as PLA15, PCL16, are multiple domain systems, are often employed as high-performance materials which can display various properties.In recent years, the development of sustainable polymers that meet the needs of consumers without destroying the environment has become an important issue due to global problems such as marine pollution, waste disposal, and global warming caused by plastics18. In addition, NMR measurements, especially low magnetic field NMR, is a method for routine material evaluations, which has produced a lot of NMR datasets19. Typical ssNMR methods are cross-polarization (CP)/magic-angle spinning (MAS) methods with elimination of linewidth broadening due to chemical shift anisotropy for high resolution. On the other hand, there are different complementary approaches to tackle complexity of polymer domain structure. 1H-static ssNMR can be applied to quantify domain mobility in terms of dynamic heterogeneity20. In addition, the use of magic-and-polarization echo (MAPE)21 and double-quantum (DQ)22 filters can determine the spectral parameters for the mobile amorphous domains with the long-time decay and the strongly dipole–dipole-coupled crystalline domains with the quickly decay, respectively.Solid-state nuclear magnetic resonance (ssNMR) spectroscopy is a powerful tool that is used to characterize the native structure, components and dynamics of solid-state samples at the atomic level, and has been increasingly applied in material sciences1H-static ssNMR measurement is useful as a measure of the kinetic nature of higher order structures, although its analysis is difficult because the spectrum is broadened and overlapped23. Therefore, application of signal deconvolution is needed to characterize structure and property of the sample. Several methods for spectral separation19, fitting and numerical simulation24 such as SIMPSON25, SPINEVOLUTION26, dmfit27, EASY-GOING deconvolution28, INFOS29, Fityk30, ssNake31, and a noise reduction method based on principal component analysis32 have been developed. So far, in NMR data analysis, signal simulation and fitting have targeted only the frequency domain or the time domain. In our previous study, we proposed signal deconvolution methods that combines short-time Fourier transform  and probabilistic sparse matrix factorization33, and non-negative tensor/matrix factorization34. In our method using STFT, by simulating the signal for both the frequency and time domain, it was possible to separate the signal related to the motility characteristics of the domain structure based on the indicators of chemical shift and T2 relaxation time. The NMR signal can be calculated by functions such as Lorentzian35, Gaussian36, and Voigt37 in the frequency domain, and by the T2 relaxation equation38 in the time domain. In addition, the difference in T2 relaxation times can be adjusted by the Weibull coefficient39. Analysis of the relaxation time of a sample's free-induction decay (FID) provides important insights into the chemical composition, structure, and mobility of the sample40.In the case of characterization of a solid-state sample with domains of rigid, intermediate and mobile types, the 41. Materials informatics, which is the emerging field, support analysis of relationships of structure and property from materials data sets42. NMR signal has potential for use as a descriptor having the structural features of the molecules contributing to their physical/chemical/biological properties43. In previous studies, a self-organization map (SOM) has been applied to tool wear monitoring44. Market basket analysis (MBA) has been applied to predict drug-drug interactions45. Bayesian optimization has been applied in real and virtual degradable experiments of bioplastics46. Generative topographic mapping regression (GTMR) has also been applied to the analysis of CP-MAS spectra to predict 13C NMR spectrum of the material in its solid-state based on its thermophysical properties34. Machine learning methods have applied for various material studies such as cloud-point engineering of polymers47, prediction of drug-polymer amorphous solid dispersion miscibility and stability48, atomic/inter-atomic properties prediction49, solubility prediction50, descriptor selection for investigating physical properties of biopolymers in hairs51, classification of the membrane materials52, prediction of crystallization tendency53, prediction of density, glass transition temperature, melting temperature, and dielectric constants of polymer9, macromolecular modeling54.The polymer domain structure has a significant influence on their macroscopic properties1H-static ssNMR spectral parameters obtained using STFT were utilized, including T2 relaxation time, frequency, and intensity. The domain structure with different mobility in the polymeric material was estimated by fitting the physical indices such as T2 relaxation time, frequency, and linewidth. In addition, using a SOM and MBA, the relationships between the estimated domain structure and the meta-information such as elements, functional group, and thermophysical property were explored.In this study, we propose a materials informatics approach to explore the structure–property relationships of polymers that combines the polymer domain modeling and the integrated analysis of polymer materials meta-information. For polymer domain modeling, 1H-static ssNMR (1H-static ssNMR) data, primary structure of the polymer, and thermophysical property data (TG/DTA/DTG). Then, frequency and time information are obtained by STFT against FIDs obtained by 1H-static ssNMR. The domain modeling method is the following. The domain components are firstly separated by fitting the obtained frequency and time information , Intermediate (Rigid) and Rigid) are calculated from Eqs. The ratios of the domain components were calculated from the volume ratios obtained from domain modeling of 71 samples of the polymer materials Fig. . Here, tT2 relaxation time and thermophysical properties of the domain components of each polymer are shown , 0.32 ms for Intermediate (Rigid), and 0.11 ms for Rigid. The weighted average (WA) of the polymer materials showed a distribution among the samples, among which PCL , differential thermal analysis (DTA), derivative thermogravimetry (DTG), differential scanning calorimetry (DSC))56. To capture the characteristics of the integrated data, clustering by SOM was performed , Intermediate (Rigid), and Rigid) shows a high lift value with the primary structure information . While the thermophysical properties  show a dominant lift value with their structure information. The lift value here is one of the indicators for correlation analysis in MBA. Figure 57. In PCL, there is a melting temperature at 66.5 °C of DTG , functional groups (aromaticity), and thermophysical properties. For input data of MBA, the domain component information calculated by the component separation method and meta-information of thermophysical properties, elements, and functional groups used is listed in Tables DTG Fig. . While tDTG Fig. a.Figure 1H-static ssNMR spectra. In our integrated analysis of domain proportions and meta-information, SOM was a useful tool for capturing trends across polymer material data. On the other hand, MBA was able to investigate the strong relationships between structure and meta-information in individual materials, including qualitative data as well as quantitative data. The relationships between mobility of domain structure and melting temperature were similar to the results shown by SOM and MBA  simulation model. Before applying this method, the FID data was phase-corrected, baseline-corrected, and inverse Fourier transformed using TopSpin . In order to calculate the ratio between the Mobile and Intermediate (Mobile) domain components obtained from the MAPE filtered spectra, and the Rigid and Intermediate (Rigid) domain components obtained from the DQ filtered spectra, the calculation errors between the four domain components and the STFT 1H-static spectra (Static) were calculated using the following equation  of the tion Eq. .6\docume61. In order to evaluate the relationships between domain structure and thermophysical properties of polymer materials, we integrated the domain component information calculated by the component separation method, 13C-CP/MAS spectra, which easily reflect primary chemical structure, and quantitative thermophysical data . To capture the characteristics of the integrated data, clustering by SOM was performed (Tables In the integrated analysis of domain proportions and meta-information of polymer materials, a SOM was produced using the R package kohonend Tables , S2. A b63. Linearly connected methylenes > 4 carbons, oxygen containing and aromaticity was set to 1 if true and 0 if false. The numeric data of domain component information calculated by the component separation method and meta-information of thermophysical properties, elements, and functional groups listed in Tables MBA was performed using the R package arules50 spectral simulation tool that automates the T2 relaxation time domain fitting, frequency fitting, and 3D domain modeling of our domain component separation method. The details of the Python program including T2 relaxation time information, frequency information, and 3D domain modeling of the present domain component separation method can be obtained at https://github.com/riken-emar/matrigica. For improving the level of accuracy of the prediction, intermediate regression models were employed when performing in-phase machine learning. In addition, we developed a website dedicated to the established domain component ratio calculation, which is freely available at http://dmar.riken.jp/matrigica/.We have created a Bayesian optimization-basedSupplementary Information.


Small molecules can either inhibit, modulate or hijack E3 function. The latter class of small molecules led to the development of selective protein degraders, such as PROTACs (PROteolysis TArgeting Chimeras), that recruit protein targets to the ubiquitin system leading to a new class of pharmacologically active drugs and to new therapeutic options. Recent efforts have focused on the E3 family of Baculovirus IAP Repeat (BIR) domains that comprise a structurally conserved but diverse 70 amino acid long protein interaction domain. In the human proteome, 16 BIR domains have been identified, among them promising drug targets such as the Inhibitors of Apoptosis (IAP) family, that typically contain three BIR domains . To date, this target area lacks assay tools that would allow comprehensive evaluation of inhibitor selectivity. As a consequence, the selectivity of current BIR domain targeting inhibitors is unknown. To this end, we developed assays that allow determination of inhibitor selectivity in vitro as well as in cellulo. Using this toolbox, we have characterized available BIR domain inhibitors. The characterized chemical starting points and selectivity data will be the basis for the generation of new chemical probes for IAP proteins with well-characterized mode of action and provide the basis for future drug discovery efforts and the development of PROTACs and molecular glues.E3 ligases constitute a large and diverse family of proteins that play a central role in regulating protein homeostasis by recruiting substrate proteins  E. coli infection (Human inhibitors of apoptosis (IAP) proteins, also often referred to as baculoviral IAP repeat-containing proteins (BIRCs) comprise a family of proteins sharing a homologous domain, called baculoviral IAP repeat (BIR) domain. The BIRC protein family consists of five Really Interesting New Gene (RING) type E3 ligases , or in trans resulting in the recruitment of two BIRC E3 ligases and ubiquitinylation followed by proteasomal degradation , an N-terminal tetrapeptide (AVPI) which inspired IAP antagonists, called “SMAC mimetics” binding to the IAP binding motif (IBM) groove of some BIR domains . Monovalradation . Monovalradation . Despiteradation .Fluorescence polarization has been widely used for the characterization of BIR domain inhibitors , but parSM-164 and GDC-0152 were purchased from Cayman Chemical (#28632 and #17810). AZD5582, BV-6, Birinapant, LCL161 and AT406 were purchased from Selleckem . A 410099.1 and UC 112 were purchased from Tocris Bioscience (#6470 and #5251). In addition, CUDC-427 was purchased from MedChemExpress (#HY-15835). Quality control of the purchased compounds was carried out by HPLC-MS mass validation which confirmed the expected molecular weight of all purchased inhibitors.For phylogenetic analysis of the BIR domains, protein sequences were obtained from UniProt and aligned using the MAFFT sequence alignment tool available in the “MPI Bioinformatics Toolkit” . The obt48H60BF2N8O7S (M + H) 941.44 Da with the found mass of 941.5 Da confirming the structure of the synthesized BRET tracer.IAP amine  (25.0 mg6 and GST tag at the N-terminus. E. coli were cultured in Terrific Broth (TB) at 37°C until an OD600 of 1.0 was reached. The culture was then cooled to 18°C and allowed to reach an OD600 of 2.5. Protein expression was induced by the addition of 0.5 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) and the protein was allowed to express overnight. Cells were harvested  and lysed by sonication  in the presence of DNase I  and cOmplete EDTA-free protease inhibitor , and recombinant protein was purified using GST-affinity chromatography in Purification buffer  and elution was carried out using Purification buffer including additional 10 mM reduced glutathione. The eluted proteins were dialysed overnight into gel filtration buffer  while the expression tag was cleaved using 1 mg tobacco etch virus (TEV) protease. The cleaved protein was passed through a HiLoad® 26/600 Superdex® 75 pg  size exclusion chromatography column and the resulting pure protein was stored in gel filtration buffer, flash frozen in liquid nitrogen and subsequently stored at −80°C for further experiments.BIRC2-BIR3 domain (260–352) was expressed as a recombinant fusion protein incorporating a HisKA and KD, respectively), and stoichiometry (n) were calculated. Measurements were carried out in technical duplicates except for the inhibitor UC-112. Data were displayed using GraphPad Prism 9.3.ITC experiments were performed using a NanoITC instrument  at 25°C in gel filtration buffer . Purified BIRC2-BIR3 protein at a concentration of 116 μM was titrated into the reaction cell containing 10 µM inhibitor dissolved in gel filtration buffer. For this protocol, the chamber was pre-equilibrated with the test compound, and the BIRC2-BIR3 domain was titrated in while continuously measuring the rate of exothermic heat evolution. The heat of binding was integrated, corrected, and fitted to an independent single-binding site model based on the manufacturer’s instructions, from which thermodynamic parameters (ΔH and TΔS), equilibrium association and dissociation constants (Tm) data were measured as described in Differences in the melting temperature (ΔKD (30 nM) in assay buffer  in a black 384-well flat bottom plate  and purified BIRC2-BIR3 domain was titrated in a concentration range from 40 μM to 600 pM. After 1 h incubation at room temperature, fluorescence polarization was measured with excitation wavelength of 485 nm and emission wavelength of 535 nm, respectively, using a Tecan Spark plate reader (TECAN). Resulting data was plotted using GraphPad Prism 9.3 software and analyzed using a nonlinear fit to calculate the probe IC50. For competition assays, 5 nM probe was added to assay buffer containing 30 nM BIRC2-BIR3 domain (2x IC50). Compounds were titrated from 20 μM to 20 nM using an ECHO 550 acoustic dispenser (Labcyte) incubated for 1 h at room temperature and subsequent read out. Data was plotted in GraphPad Prism 9.3 and analyzed using a nonlinear fit [equation: Y = 100/(1 + 10^((X-LogIC50)))] for IC50 determination. KI calculation was performed using the Cheng-Prusoff equation (KI = IC50/(1+([R]/KD))] with KD = 15 nM, [R] = 30 nM and IC50 determined in each assay.For the complementation assay, the fluorescently labeled SMAC probe (AVPIAQKSEK-K(5-FAM)-NH2) was diluted to 2-fold equation  . Competitive displacement data were then graphed using GraphPad Prism 9.3 software using a normalized 3-parameter curve fit with the following equation: Y = 100/(1 + 10^(X-LogIC50)).The assay was performed as described previously . In brieD, app  were pipThe human proteome encodes for 16 BIR domains that are present in 8 different BIRC proteins. The first objective towards a selectivity platform was to analyze the similarity of the different BIR domains and we therefore aligned the individual BIR domains based on their amino acid sequence homology. The resulting tree is depicted in in vitro biophysical assays including fluorescence polarization (FP) assay, differential scanning fluorimetry (DSF) and isothermal titration calorimetry (ITC) were set up for initial characterization of SMAC inhibitor binding. Since thus far most medicinal chemistry approaches have targeted the BIR3 domain of BIRC2 (BIRC2-BIR3), this domain was chosen as a representative BIR domain for a comprehensive analysis. Each of the chosen assays complements the other in its ability to characterize the binding of a chemical compound to the BIR domain. Using these assays, 10 diverse and commercially available inhibitors were selected  assays have often been the first choice for  14.9 nM . Overall50 values between 1.8 ± 0.7 nM (SM-164) and 8.4 ± 0.7 nM (BV-6) with no discernable difference between monovalent and bivalent compounds derived from the same SMAC mimetic inhibitor. As the nonlinear fits did not reach their respective lower plateaus due to the high affinity of the binders, below the protein concentration used in the assay, these numbers represent approximations  was performed on the BIR3 domain of BIRC2. First, we determined the melting temperature of the native BIR3 domain of BIRC2, which had a mid-point of the unfolding transition at ∼74°C. The assay was run in the presence of DMSO to exclude DMSO influences towards melting temperature differences when testing small molecules . Despiteied BIR3 .As a third biophysical method for characterization of the literature compounds, isothermal titration calorimetry was performed. This method allowed the determination of accurate binding constants in solution without the need of tracer or dye molecules and additionally provide insight into the thermodynamic properties of the inhibitor interaction and the binding stoichiometry . GeneralE. coli. Individual BIRC1-BIR domains however were successfully obtained, indicating potential toxicity of the full-length BIRC1 towards E. coli despite using a T7/CMV promoter which should not be transcribed by the used strain. We were not able to obtain a BIRC6 NanoLuc expression construct due to the size of over 14500 nucleotides and the lack of a suitable cDNA template. BIRC5 full-length protein only contains an additional helix in addition to the BIR domain and the full-length sequence was therefore considered as a single BIR domain construct.As a critical input for the family-wide assessment of compounds in a cellular environment, we established NanoBRET assays for full-length BIRC proteins as well as individual BIR domains. All constructs were cloned using a NanoBRET-vector harboring an N-terminal NanoLuc. Unfortunately, BIRC1 and BIRC6 constructs were not obtained. BIRC1 full-length cloning failed, resulting in accumulation of mutations when transformed into KD of 21.1 ± 10 nM measured by ITC. The tracer was not only tested on BIR domains expected to bind SMAC mimetics, but all successfully cloned and expressed BIR domains.Since the development of SMAC mimetics has been based on the AVPI motif of SMAC, all compounds were expected to bind to the IBM groove. Based on this, a pan-BIRC tracer was synthesized, using LCL161 as parent compound , which hKD of lower than 250 nM was measured with z’ values of 0.6–0.7, indicating good assay quality. BIRC8 had lower affinity for the tracer. Nevertheless, a surprisingly high assay quality with a z’ of 0.8 and an assay window of 42.9 was obtained, resulting in a high-quality assay panel of full-length BIRC proteins  and BIRC1 has been suggested to have an SMA modulating role  but also an ungrouped BIRC1-BIR[2] and a fourth “BIR4” domain group , the functional importance of which still needs to be determined . Both, Bing role . HoweverThe E3 BIRCs, BIRC2-4, together with BIRC1-BIR1 encode a BIR1 domain. The BIR1 domains of BIRC2 and BIRC3 do not possess the IBM grove found in BIR3 and BIR2 domains, but instead interact with TNF Receptor-Associated Factor 2 (TRAF2) as part of the TNFα signaling pathway . This pain vitro affinity validation. Data obtained by the fluorescence polarization assay showed low nanomolar KI values for the different SMAC mimetics, which generally agreed with published data. In our assay, most of the compound titrations showed 2 data points (2 and 5 nM) with significantly higher mP values with respect to the residual data points, indicating, that the ‘assay wall’ is located between the used peptide tracer concentration and the peptide tracers KD. Since the majority of calculated KI values were smaller than 5 nM, and therefore smaller than the detection limit, the general conclusion for of the FP assay is that all SMAC mimetics have shown extraordinary high affinities of smaller than 10 nM towards BIRC2-BIR3 proving the FP assay to be applicable for interaction determination of compounds with lower affinity while the non-SMAC mimetic UC-112 has shown no interaction at all. UC-112 has been reported as an inhibitor for BIRC5 (Survivin) based on molecular modelling and cellular data monitoring apoptosis  were found to be well cell membrane permeable as indicated by their comparable affinity to the monovalent SMAC mimetics. However, biological activities such as cell viability, degradation of c-IAP1 and activation of noncanonical NF-kB pathway may well differ from the measured on-target effect for individual BIRCs, in particular given the promiscuous nature of the compoundsInterestingly, two compounds were identified that showed preferential binding to BIRC7. AZD5582 is more than 10-fold more potent for full-length BIRC7 compared with any of the remaining full-length proteins . This shFuture structural studies on the single BIRC7-BIR domain bound to AZD5582 and CUDC-427 may provide insights for the development of BIRC7 selective inhibitors. We also observed some additional domain selectivity: AZD5582 and A 410099.1 showed significantly higher affinity towards BIR2 domains compared to other inhibitors. Some of the investigated compounds, such as AT406, SM-164 and AZD5582 bound to the BIRC1-BIR[2] domain, which is an atypical BIR domain that has not prior been shown to interact with SMAC mimetics, expanding the target scope of SMAC mimetics outside the E3 ligases family members.In this work, a BIRC family-wide selectivity screening platform was established. This selectivity toolbox comprises all full-length E3 ligases, the non-E3 ligase BIRC5 and 7 of the 14 single BIR domains of this family. The here presented selectivity panel therefore consists of BIRC1 (BIR[2]), BIRC2 , BIRC3 , BIRC4 (FL), BIRC5 (FL), BIRC7 (FL and BIR) and BIRC8 (FL and BIR). Hence, all five E3 ligases can now be used for selectivity profiling of putative inhibitors in living cells. Due to the similarities of their BIR3 domains, selectivity of the BIRC E3 ligases may be difficult to achieve. This setting allows to evaluate selectivity profiles of E3 ligands, novel chemical probes and PROTACs with the possibility of domain specific screening campaigns.Strikingly, most of the tested compounds did not show any selectivity within the BIRC family, including clinical candidates like LCL161, Birinapant and AT406 (Xevinapant) defining them as pan-BIRC inhibitors. Due to the general involvement of BIRC proteins in a diverse range of pathways, a pan-BIRC inhibitor could lead to unwanted effects which could be avoided by a development of selective BIRC inhibitors targeting only the BIRC proteins involved in the specific signalling pathway. Since BIRC1 has been shown to play a role in response to microbial infections , the inh