The nutritional and palatability relevance of bread prepared with soy flour was examined. There are a few effective nutritional measures that combine palatability, convenience, and functionality in the suppression of muscle loss (contributing to the improvement and prevention of sarcopenia). Therefore, in the present study, we attempted to produce bread using soybeans, which are rich in amino acids involved in the synthesis and degradation of skeletal muscle proteins. Rice flour was also used to avoid gluten intolerance. The bread was baked in an automatic bread maker, and the rheological properties of its breadcrumbs were determined using a creep meter. We found that a 70 g slice of soy bread satisfied approximately one-fifth of the daily nutritional requirement for leucine. Although soy decreased the specific volume of bread by preventing starch construction, the use of preprocessed rice flour recovered the volume, and corn starch improved the taste. We propose that the addition of soy bread to the daily diet may be an effective protein source. The 2022 annual report of the World Health Organization (WHO) shows that both healthy life expectancy (HALE) and overall life expectancy (LE) have increased in developed countries . HALE waSarcopenia is defined as an age-related decline in skeletal muscle mass and physical function (strength) and is associated with an increased risk of falling and disability ,3,4,5,6.Certain amino acids play a more prominent role in protein metabolism, rendering them ideal targets for inclusion in a protein-rich food source. The mechanistic/mammalian target of rapamycin (mTOR) is an evolutionarily conserved serine/threonine kinase that is known to be a master regulator of cellular metabolism. mTOR stimulates anabolic processes, such as protein synthesis while simultaneously inhibiting autophagy, which includes protein degradation. Leucine, a branched-chain amino acid (BCAA), activates mTOR signaling more efficiently than other amino acids . In addiClock and Bmal1, and by BCAA ingestion [Another important factor affecting efficient protein synthesis is the timing of its intake, which can influence muscle hypertrophy. In particular, a high-protein (BCAA) breakfast is more efficient in establishing muscular hypertrophy than an equal distribution of protein intake across the day or a higher intake at dinner. This time-dependent effect is affected by clock genes, such as ngestion . Howeverngestion .The current study seeks to evaluate formulations for producing protein-rich bread that can assist in efficient protein synthesis, which in turn can prevent the loss of skeletal muscle mass. Although wheat-based bread is commonly consumed for breakfast, it contains gluten that can induce gastrointestinal discomfort. Zonulin is a physiological modulator of intestinal tight junctions that leads to a leaky gut. Gliadin, a protein component of wheat gluten, activates zonulin signaling, which increases intestinal permeability to macromolecules ,20,21,22The specific aims of the study were to investigate the effects of sub-ingredients on the quality of bread baked with rice flour and soy flour supplements as an initiative to increase protein consumption by humans and thus contribute to the prevention of sarcopenia. Our results showed that although soy flour supplementation to rice bread reduced the specific volume , the volume was recovered using preprocessed rice flour and cornstarch, which participate in the fermentation process.The ingredients of the bread comprised rice flour , soy flour , wheat flour , cornstarch , starch , sugar , salt , preprocessed rice flour , dry yeast , and canola oil .The bread samples were prepared using an original method designed by us. The components of each base formulation (baker’s %) were rice flour (100), water (82), sugar (5), salt (2.1), dry yeast (1.5), and canola oil (5.7). For wheat bread, wheat flour was used instead of rice flour. The total amount of flour was adjusted when soy flour, preprocessed rice, and cornstarch were added to the formula. The bread was baked using the rice bread program or wheat bread program of an automatic bread machine . The baked bread loaves were cooled at room temperature (25 ± 2 °C) for 1 h before performing various measurements.After cooling, the weight and volume of the bread loaves were measured. The volume was determined based on the rapeseed displacement method . The mass was measured using a digital scale (TANITA CORPORATION). The specific volume was calculated as the ratio of volume to weight (mL/g).A creep meter  was used to determine the rheological properties of the breadcrumbs. The parameters for measuring rupture characteristics included sample size, 30 × 30 × 20 mm; plunger, wedge form ; measurement strain rate, 100%; and compression speed, 1.0 mm/s. Conditions for the measurement of bread texture included sample size, 30 × 30 × 20 mm; plunger, circular form (12 mm diameter); measurement strain rate, 50%; and compression speed, 5 mm/s. Each experiment was performed at least in triplicate, and averages were obtained for the data.Protein and amino acid contents of bread types  were analyzed using the combustion method and HPLC , respectively, by the Japan Food Research Laboratories. For each bread sample, 0.31–0.39 g and 0.2–0.3 g were used for the protein and amino acid analyses, respectively. The samples were dissolved in 20 mL of 20% hydrochloric acid containing 0.04% 2-mercaptoethanol. After hydrolysis , the samples were quantitated and prepared for HPLC measurement. Ninhydrin was used as the reaction reagent, and tryptophan was measured by HPLC (fluorescence detection), whereas other amino acids were measured by post-column derivatization. Each sample was analyzed twice.The water absorption of each flour sample was determined using the technique developed by Matsuki et al. . Seven hSensory analysis of the bread was carried out according to Japanese Industrial Standard   by a panp < 0.05 was considered statistically significant.All results were obtained from at least three separate experiments. Statistical differences were analyzed via one-way analysis of variance (ANOVA) and Tukey’s range test for multiple comparisons using IBM SPSS . The results are expressed as mean ± standard deviation (SD), and To determine the ideal amount of soy flour for baking rice bread, we investigated 15, 25, 35, 50, and 75% replacements of soy flour with rice flour. The specific volume is an important indicator of the technological quality of bread and is used to express the technological aptitude of a formulation for bread production [Saito et al.  reportedNext, the most appropriate quantity of pregelatinized rice flour was determined. As shown in The amino acid content of the different bread types is shown in The effects of pregelatinized rice flour and cornstarch on the baking properties of soy bread are shown in 2 gas retention [2 during mixing and proofing. Furthermore, soy proteins may interact with amylose and starch granules through non-covalent bonds, thereby reducing viscosity and interfering with the association of hydrogen bonds between the starch molecules [The volume and specific volume of bread depend on the retention of gas by the matrix during fermentation and affect bread quality . Next, wetention . Consistetention  and riceetention ,31,32 reetention  reportedetention  observedolecules . MizuhocBread quality is also affected by the water absorption of its constituent components. Accordingly, we examined whether there was a difference in water absorption between each bread type. In comparison with the control , water absorption significantly increased in the soy bread and increased further by the use of pregelatinized rice. In contrast, the addition of cornstarch returned water absorption levels to those of the control .2 gas emitted during the fermentation process, thereby improving bread quality in terms of specific volume. Soy absorbs water but does not influence batter viscosity and only increases its weight; this may be due to the prevention of the pregelatinizing action of rice flour, thereby leading to a reduction in bread-specific volume. Cornstarch returned the increase in adhesiveness observed for pregelatinized rice flour to that of the control. Cornstarch suppressed water absorption and attenuated excess batter adhesiveness, resulting in an appropriate viscosity of the bread.Next, we examined whether water absorption influenced batter adhesion and whether pregelatinized rice flour increases the absorption of water, resulting in the thickening of the batter. 2 gas in the batter, thus leading to a loss of specific volume in the bread.In summary, soy disrupted the cross-linking of rice starch (which determines adhesiveness) and interfered with the retention of COp > 0.05) differences were observed between the 35% soy flour replacement bread , the soy rice bread with preprocessed rice flour (F2), and the soy rice bread with preprocessed rice flour and cornstarch (F3), the chewiness and stickiness of F3 tended to be lower than those of F1 or F2. Moreover, F3 tended to have sponginess and firmness qualities similar to those of F1, which were superior to those of F2. In the sensory evaluation, the following comments were made on the soy rice bread with preprocessed rice flour: “I felt that F2 is like a mochi, F3 has no bad taste and aroma, and F3 is dry as compared with F2”. It is important to note that we chose to include a panel of young people for this preliminary study to ensure safety, as textures similar to that of rice cake can readily cause dysphagia in elderly individuals. Thus, further sensory analysis is needed with a panel comprised of elderly people, and additional studies are required to assess the inhibitory effect of these breads on skeletal muscle declension.The results of the descriptive sensory analysis performed on the three breads are shown in An increasing number of people exhibit symptoms of gluten intolerance. The most common wheat flour substitutes for gluten-free bread are rice flour, which is mainly composed of carbohydrates and has a low protein content. Numerous studies have attempted to improve not only the quality but also the nutritive values of gluten-free bread by increasing its protein contents using plant- and animal-derived components. In this study, we evaluated the formulations for producing protein-rich bread. We used soy flour owing to its efficient protein synthesis activation and proteolysis inhibition effects. Although several studies have added soy flour to wheat or rice flour bread, the bread quality has remained uninvestigated. We observed a deterioration in the specific volume of soy bread; however, we attempted to improve the soy bread quality. Although there have been several studies on the addition of soybean flour to wheat and rice flour breads, improvements in bread quality have not been explored. We found that soybean inhibits the cross-linking of rice starch, which determines adhesion, and inhibits carbon dioxide retention in the batter, thereby reducing the specific volume of the bread. This can be applied to the production of gluten-free soy bread, for example, by preparing bread with improved cross-linking of rice starch, i.e., by either inhibiting or not inhibiting cross-linking. In this study, we used preprocessed rice flour to improve the cross-linking of rice starch, but we believe that this is possible with other materials as well, which are currently considering. Although we developed soy bread that may increase protein supply, it is necessary to continue investigating other formulations to improve nutritional aspects, quality, taste, and costs, including the inclusion of additional ingredients or changes to the recipe, while also confirming whether soy bread has beneficial effects on skeletal muscle in elderly people.2 in the dough. However, the lost volume was recovered using preprocessed rice flour, and the taste was improved using cornstarch (which inhibited the excess of adhesiveness produced by preprocessed rice flour). We propose that soy bread has the potential to become an effective countermeasure against skeletal muscle declension. Soy bread is expected to enhance the nutritional value and quality of bread. Based on the findings, soy bread is a potential source of protein in the daily diet. However, further research is required to improve the nutritional aspects, baking quality, and cost of soy bread, while also ensuring optimal taste and minimal dysphagia in elderly individuals.This study considers a nutritional approach to contribute to the prevention and improvement of skeletal muscle decline in the future. We attempted to produce quality soy bread as a breakfast meal option for the elderly, considering its provision of amino acids at an appropriate time to allow efficient muscle protein synthesis. Since the sustainability of use is a key factor to success, we considered bread taste and avoided gluten intolerance issues by using rice flour in analyzing bread quality. Nutritionally, one 70 g slice of soy bread delivered approximately 19% of the total BCAA and daily leucine requirements (60 kg body weight). In terms of baking quality, soy significantly decreased the specific volume of bread by inhibiting starch cross-linking that retains CO


MS7.4 earthquake obtained by the method proposed in this paper, was compared with that obtained from the only InSAR measurements obtained using a multi-satellite and multi-technology approach. The results showed the difference in root-mean-square errors (RMSE) of the integration and GNSS displacement was 0.98 cm, 5.64 cm, and 1.37 cm in the east–west, north–south and vertical direction respectively, which was better than the RMSE of the method using only InSAR and GNSS displacement, which was 5.2 cm and 12.2 cm in the east–west, north–south, and no vertical direction. With the geological field survey and aftershocks relocation, the results showed good agreement with the strike and the position of the surface rupture. The maximum slip displacement was about 4 m, which was consistent with the result of the empirical statistical formula. It was firstly found that the pre-existing fault controlled the vertical deformation on the south side of the west end of the main surface rupture caused by the Maduo MS7.4 earthquake, which provided the direct evidence for the theoretical hypothesis that large earthquakes could not only produce surface rupture on seismogenic faults, but also trigger pre-existing faults or new faults to produce surface rupture or weak deformation in areas far from seismogenic faults. An adaptive method was proposed in GNSS and InSAR integration, which could take into account the correlation distance and the efficiency of homogeneous point selection. Meanwhile, deformation information of the decoherent region could be recovered without interpolation of the GNSS displacement. This series of findings formed an essential supplement to the field surface rupture survey and provided a novel idea for the combination of the various spatial measurement technologies to improve the seismic deformation monitoring.Three-dimensional deformation is an important input to explore seismic mechanisms and geodynamics. The GNSS and InSAR technologies are commonly used to obtain the co-seismic three-dimensional deformation field. This paper focused on the effect of calculation accuracy caused by the deformation correlation between the reference point and the points involved in the solution, to build a high-accuracy three-dimensional deformation field for a detailed geological explanation. Based on the variance component estimation (VCE) method, the InSAR LOS, azimuthal deformation, and the GNSS horizontal and vertical deformation were integrated to solve the three-dimensional displacement of the study area in combination with the elasticity theory. The accuracy of the three-dimensional co-seismic deformation field of the 2021 Maduo  The three-dimensional deformation field caused by any geological hazard is an important parameter in the study of crustal movement characteristics, which implies certain geodynamics. Accurately acquiring the complete regional three-dimensional crustal deformation field is an important research direction for the dynamics of regional crustal deformation and understanding its geohazard process. The rapid development of geodetic measurement technologies, such as InSAR (Interferometric Synthetic Aperture Radar) and GNSS , provides an opportunity to acquire an accurate regional three-dimensional crustal deformation field. The InSAR has the advantages of all-weather, all-day, high-spatial resolution, while it only provides one-dimensional deformation information, so that the monitoring of true crustal deformation dynamics is restricted. The GNSS can provide high-precision three-dimensional deformation information , but itsIntegrating the advantages of these two observations to obtain high-accuracy and high-spatial-resolution three-dimensional deformation, is a subject that has been explored. Since Guemundsson  proposedGuglielmino  proposedThe high construction cost and strict setting requirements determine the uneven distribution density of GNSS stations. In many cases, only InSAR was used to obtain three-dimensional deformation ,28,29,30MS7.4 earthquake in Qinghai, China, which occurred in the Bayan Har block as an example, we collected more than 100 GNSS stations in the surrounding area of interest, and the InSAR co-seismic one-dimensional deformation. Based on the elasticity theory [Taking the 2021 Maduo y theory , we estay theory ,40,41,42MS7.4 earthquake occurred in Maduo County, Luozhou, Qinghai, China on 22 May 2021, whose epicenter located at 34.59° N, 98.34° E, and the source depth was 17 km , which was the largest earthquake after the 2008 Wenchuan MW 7.9 earthquake in China. The Maduo MS7.4 earthquake occurred on the Bayan Har block of the Qinghai–Tibet Plateau secondary tectonic block, 70 km south of the Toso Lake section of the East Kunlun Fault. The Bayan Har block had a very complex tectonic background, and was surrounded by the Ganzi–Yushu–Xianshuihe Fault to the south, the Longmenshan Fault to the east, the East Kunlun Fault to the north, the Maergaichaka Fault and the Altyn Fault to the west. The southern side of the Bayan Har block was pushed by the Indian plate to the northeast and was blocked by the Tarim Basin, which caused material flow and formed the uplift deformation on the Qinghai–Tibet Plateau. Due to the blockage of the eastern Erdos block and the Sichuan Basin, the Qinghai–Tibet Plateau as a whole continued escaping to the southeast, and the geotectonic movements were active and easily induced large earthquakes in the block. In recent years, many strong earthquakes have occurred on the Bayan Har block, such as the 2001 Kunlun Mountain Pass West M8.1 earthquake, the 2008 Wenchuan M8.0 earthquake, the 2010 Yushu M7.1 earthquake, the 2013 Lushan M7.0 earthquake, and the 2017 Jiuzhaigou M 7.0 earthquake, which indicated that the block was in a strong active state. It was necessary to pay more attention to geological hazards in this area. The 2021 Maduo MS7.4 earthquake occurred on a secondary fault that was approximately parallel to the East Kunlun Fault [MS7.4 earthquake and the used data in the area of interest.An un Fault . HoweverMS7.4 earthquake had been researched by various means [So far, the seismogenic structure and deformation characteristics of the Maduo us means ,51,52,53us means ,54, whicThe GNSS stations from different organizations provided the research conditions for the discrete three-dimensional deformation in the Qinghai–Tibet Plateau. However, the regional co-seismic three-dimensional deformation could not be solved simply by interpolating GNSS data. As a complement for the low spatial resolution of GNSS stations, InSAR measurement could be used to obtain high spatial resolution one-dimensional deformation of the study area. Using the GAMIT/GLOBK software, the time series of high accuracy GNSS three-dimensional deformation could be obtained before and after the earthquake. This paper applied the continuous GNSS observation stations from different sources, including the Crustal Movement Observation Network of China, the provincial earthquake agency, enterprises, and more than 200 IGS stations uniformly distributed worldwide . Firstly, all the stations used were loosely constrained, and their daily solutions by the GAMIT software were obtained, where the baseline solution used the RELAX mode, the satellite elevation cutoff angle was 10°, the atmospheric zenith wet delay was solved every 2 h, and mapped to the satellite by the GMF mapping function. All files, including the receiver antenna file, the satellite antenna file, the receiver and satellite antenna phase center models, the solid tide model, the solar ephemeris table, the lunar ephemeris table, and the ocean tide model, were updated. The daily solutions were combined with the calculated global distribution of 200 IGS stations. then, we chose some global, even, stable, and self-consistent IGS stations as constraints, and performed a seven-parameter transformation to tie the above solution to the International Terrestrial Reference Frame 2014 (ITRF14) by the GLOBK software. We could obtain the accurate three-dimensional coordinates time series under the ITRF 2014.To avoid the after-slip effect and increase the GNSS measurement reliability, the co-seismic displacement of GNSS stations before and after the earthquake was chosen based on the principle that the ratio of displacement to error was greater than 1. Some data after the earthquake was removed, so we used selected data, including 3 days before the earthquake and 1~3 days after the earthquake, to calculate the co-seismic deformation. The station time series were averaged, respectively, and the differential before and after the earthquake, the errors in each direction could be obtained by the error propagation law. In InSAR was the space-to-earth observation combining synthetic aperture radar (SAR) technology with interferometric technology. The SAR satellites are in an almost polar orbit around the north and south poles of the Earth, thus it was very insensitive to the north–south (NS) deformation of InSAR LOS direction, while the azimuthal direction could make up for the shortcoming of the InSAR LOS direction. The DInSAR technique could acquire the LOS deformation, and the MAI and POT techniques could acquire the LOS and azimuthal deformation, in which the azimuthal accuracy of the MAI was significantly better than that of the POT in areas with higher coherence, while the POT could resist decoherence and obtain the fault trace, which all belong to the InSAR measurement.The SAR satellite provided two different geometry SAR images by ascending and descending orbits, respectively. The research shows ,57 that https://scihub.copernicus.eu/dhus/#/home, accessed on 12 July 2021) by registered users, and the corresponding precise orbit file can be downloaded freely about 21 days after measurement on the other website . We downloaded the Sentinel-1A/B interferometric wide (IW) swath mode single look complex (SLC) data before and after the 2021 Maduo MS7.4 earthquake, as well as the precise orbit files at the corresponding time. The specific information is shown in The European Space Agency (ESA) launched two C-band SAR satellites, the Sentinel-1A satellite, and Sentinel-1B satellite, in April 2014 and April 2016, respectively. The revisit period was 12 days for a single satellite, while the revisit period could be shortened to 6 days for two satellites. The Sentinel-1 observation data could be downloaded freely from ESA official website H=ε+By bringing the Equation (2) into Equation (1), it could be expressed asIn which, the unknown matrix was expressed as follows.The coefficient matrix was as follows.Then,For the GNSS measurement, the GNSS stations discrete co-seismic three-dimensional deformation can be directly obtained by displacement difference before and after an earthquake. It can be expressed as follows.For the InSAR measurement, only one-dimensional deformation could be obtained in the azimuthal or LOS direction, and needed to be decomposed into three-dimensional direction by a certain formula. The relationship between three-dimensional and one-dimensional displacement of the same point was as follows.In whichAccording to Equations (6)–(8), the relationship between the observations and the calculated values was as follows.The error equations could be listed as follows.The research showed that  the numbfunction  between When more than 200 pixels were involved in the solution, the RMSE tended to be steady . We lineThe heterogeneous points near the fault must be considered in the process of adaptively adjusting the window sizes, due to the deformation characteristics being different on both sides of the earthquake fault. To search heterogeneous points around the reference point and remove them, we obtained the specific earthquake fault trace and the boundary of the heterogeneous points, due to the POT being insensitive to decoherence. It was different from the empirical approach that all GNSS stations were involved in the solution; we chose the GNSS stations within the correlation distance in order to ensure the deformation correlation between the used observations and the reference point. It meant that the number of the GNSS stations and the InSAR pixels involved in the solution were variable pixel by pixel.After determining the observations involved in the solution by adaptively adjusting the window size, we could establish the error equation. We considered weighting reasonably for the different observations.The observations were classified according to their measurement accuracy. The GNSS observations were weighted as two types of observations, due to the different measurement accuracy in the horizontal and vertical directions. InSAR observations were weighted according to the directions and the orbit from different measurement technologies. Each type of observations was considered to be independent. Firstly, the prior variances reciprocal was as the initial weights. For the GNSS measurement, the prior variances were the error square in the corresponding directions. For InSAR measurement, the prior variances were calculated by using the pixels within the adaptive window size base on the ergodicity assumption; the formula is as follows.After estimating the unit weight variance of each type of observation by the VCE method, the corresponding new weight was given by iterating the initial weight, that was as follows.2. That is, the end-of-iteration condition was satisfied In the weighting iteration process, the difference between the different types of unit weight variance should be less than a convergence threshold In this paper, the three-dimensional surface deformation field in the hundreds of kilometers along the seismic fault near the epicenter was obtained by effectively integrating the GNSS and InSAR measurements. The strike of the earthquake fault was NWW and locally near EW. Since the angle between the strike of the earthquake fault and the east–west direction was small, the east–west deformation was dominant in the earthquake fault deformation. We considered that the east–west component result could indicate the earthquake fault movement mode. As shown in MS7.4 earthquake indicated [The field scientific investigation of the 2021 Maduo ndicated  that no We overlaid the main surface rupture of the geological mapping with the three-dimensional deformation field. In og(Dmax) . The deformation range in the south of the earthquake fault were significantly larger than that in the north of the earthquake fault, which may indicate that the primary power source causing this earthquake was on the south of the earthquake fault, which was consistent with the knowledge that the Qinghai–Tibet Plateau has been driven northward by the Indian plate. MS8.1 earthquake [rthquake . Accordirthquake ,53. The rthquake . Where trthquake .MS7.4 earthquake were from the epicenter to the east and to the west and had obvious broom-shaped bifurcations at both ends of the rupture. This phenomenon was a typical manifestation of energy release and conformed with the principles of seismology. However, it was a relatively rare deformation feature within the Bayan Har block.The surface ruptures caused by the Maduo An important finding was that the vertical component had special deformation characteristics at the west end of the main surface rupture, called the near east–west main surface rupture. The boundary between the uplift and subsidence was located on the north side of the near east–west main surface rupture See , while tCombining the deformation result with the field survey, the above analysis results proved the reliability of the calculated three-dimensional deformation, and illustrated the importance of the three-dimensional deformation field for the fine interpretation of the surface rupture and the fault kinematics characteristics.While the InSAR could not directly provide three-dimensional deformation, the GNSS three-dimensional deformation played an important role in solving the regional three-dimensional deformation field. Compared to the GNSS measurement discreteness, the InSAR measurement was important in spatial scale. The integration of GNSS and InSAR complement each other to maximize their advantages. The differences in root-mean-square errors (RMSE) between calculated three-dimensional deformation and the corresponding GNSS stations were 0.98 cm, 5.64 cm, and 1.37 cm in the east–west, north–south, and vertical directions. The accuracy was better than that of only InSAR measurement, whose RMSEs were 5.2 cm and 12.2 cm in the east–west and north–south directions, but none in the vertical direction . The accFor different types of observations, the calculation precision of the variance component estimation (VCE) weighing was better than the weighted least squares (WLS), which used the observation prior variance as the weight with some uncertainty assumptions, which made it difficult to accurately reflect the observation weight. We took the reciprocal of the observation prior variances within the adaptive window size as the initial weights, then carried out several iterations based on the VCE to obtain the reasonable weights of the observations. The analysis result verified that it was feasible to obtain the high accuracy three-dimensional deformation field by the VCE weighting.M7¾ earthquake and the 2021 Maduo MS7.4 earthquake.The Bayan Har block located in the central and eastern regions of the Qinghai–Tibet Plateau is one of the strongest seismically active regions in China nowadays . Its souThe three components interpretation for the co-seismic deformation was helpful to realize the kinematic processing of the seismogenesis structure, and provided the reference for understanding the geological structure and the geodynamics in this region. The three-dimensional deformations showed that the seismogenesis fault had a typical left-lateral strike-slip movement, and the two ends of the main surface rupture had obvious displacement and strike deflections. It was speculated that the fault was blocked by some hard material (obstacle body) during the strike-slip process, which led to the main surface rupture strike southward deflection at the west end and formed two branches at the east end. This was more conducive to energy release in some way. The energy dispersion led to surface distributed deformation .The east–west deformation dominating the NWW strike-slip displacement, had a good indicator of the regional geodynamic environment. The active source of this earthquake might lie in the south of the earthquake fault, which was consistent with the fact that the Qinghai-Tibet Plateau is continuously pushed by the southern Indian plate to the Eurasian plate . The norThe vertical deformation in both sides of the main surface rupture See c,f beingMS7.4 earthquake caused at least three different strike ruptures. As field investigation from Li et al. [Intuitively, the southward deflection at the west end and the bifurcation rupture at the east end of the main surface rupture showed that the 2021 Maduo i et al.  and Pan i et al.  showed, i et al.  used thei et al. , who beli et al. .The continuous surface rupture was the most obvious in the south of the Eling Lake. The rupture’s geometric distribution was consistent with the bifurcation phenomenon of the aftershock sequence relocation  see Fig, which mMS8.1 earthquake [MS7.1 earthquake [The surface rupture and deformation characteristics of the Maduo earthquake were significantly different from those of several strike-slip earthquakes that had occurred in the Qinghai–Tibet Plateau in the past, and this indicates the complexity of the earthquake. For example, the main surface rupture of the 2001 Kunlun Mountain Pass West rthquake , locatedrthquake , which oMS7.4 earthquake [In the surface rupture, we could see some left-stepping tensional cracks and pull-apart basins; the mechanism of these phenomena was caused by the Riedel shear. The major strike-slip faults, which may be thousands of kilometers long and tens of kilometers wide, are in domains of simple shear, and the displacement can be monitored in hundreds of kilometers along a fault. Within the domain of simple shear, the most recently active strand might be a zone of active faulting that was only a few meters wide. The simple shear had a donoclinic symmetry of strain because it was rotational, and a greater variety of structure forms in simple shear than in pure shear . The en rthquake . The focrthquake ,85,86,87The calculated three-dimensional deformation field was in agreement with several geological field surveys ,95,96,97MS7.4 earthquake was a typical left-lateral strike-slip earthquake; the revealed kinematic characteristics of the seismogenic fault showed that there was a relatively obvious dip-slip component at the ends of the rupture.Combining the surface rupture field investigation with the regional three-dimensional deformation field by integrating the GNSS and InSAR, we considered that the 2021 Maduo In the inversion of the three-dimensional co-seismic deformation field, we also made the important discovery that there was the obvious left-lateral strike-slip deformation but no vertical deformation at the west end of the main surface rupture, while the pre-existing fault dominated the vertical deformation. This discovery was an important supplement to the co-seismic deformation characteristics obtained from the field investigation. It provided an important reference for further exploring this earthquake’s deformation mechanism and regional geodynamics.The deformation correlation between the reference point and the points involved in the solution was fully considered, and it was taken as the decorrelation distance range for selecting the points used. An adaptive method was proposed to obtain a high accuracy regional three-dimensional deformation field. Taking the 2021 Maduo MS7.4 earthquake as an example, based on the elasticity theory, we established the accurate regional co-seismic three-dimensional deformation field by effectively and rationally integrating GNSS and InSAR, and gave the crustal dynamics and geological explanation of the three-dimensional co-seismic deformation field. The conclusions could be drawn as follows.(1)The accuracy obtained by integrating GNSS and InSAR increased by 81.2% and 53.8% in the east–west direction and north–south direction, respectively. Additional constraints were helpful to obtain a high accuracy regional three-dimensional deformation field, such as increasing the GNSS stations, considering deformation correlation, removing heterogeneous points around the reference points, and the variance component estimation weighting.(2)The spatial position and the left-lateral strike-slip motion of the earthquake were consistent with those of the Jiangcuo section of the NWW strike of the Kunlun Mountain Pass–Jiangcuo fault in the south of the East Kunlun Fault. The mechanism of the left-lateral left-stepping was caused by the Riedel shear, which dominated the formation and evolution of strike-slip faults.(3)The maximum slip of the earthquake was about 4 m, which was difficult to investigate in the geological field survey due to the lack of reliable markers, and the slip was consistent with the empirical statistical results.(4)The ruptures started from the epicenter to both ends and formed bifurcations, which were rarely found in the same type of previous earthquakes that occurred in the Qinghai–Tibet Plateau.(5)The pre-existing fault controlled the vertical deformation at the north side of the near east–west surface rupture, which only dominated the east–west strike-slip. The three-dimensional deformation was consistent with the surface rupture trace. This provided direct evidence for the theoretical hypothesis that large earthquakes can not only produce a surface rupture on seismogenic faults, but also trigger pre-existing faults or new faults to produce surface ruptures or weak deformation in areas far from seismogenic faults.We recognized that the new seismic surface rupture did not necessarily completely control the distribution of seismic deformation. For example, in the western end of the surface rupture of the Maduo earthquake, the strike-slip deformation was distributed along the new near east–west surface rupture, while the vertical deformation was controlled by the pre-existing structure in the northwest direction.The deformation distribution range in the south of the fault was wider than that in the north, which might indicate that the physical properties of the geological bodies were different on both sides of the fault. The northward extrusion of the Indian plate led to the formation of large strike-slip faults in the Qinghai–Tibet Plateau, and these faults constituted the boundary of the active block. The epicenter was located on the south of the East Kunlun fault, which was the northern boundary of the Bayan Har block. The co-seismic deformation field indicated that the thrusting effect from the southern Indian plate was transmitted to the epicenter. The block in the south of the earthquake fault moved faster in the SEE direction than the block in the north. In future, we would obtain the displacement distribution on both sides of the surface rupture through a series of deformation profiles, and compare it with the field investigation to further discuss the changing trend of deformation and its mechanism. In addition, the relationship between the earthquake fault and the East Kunlun fault, and the relationship between the new rupture and the pre-existing fault are topics to be further studied.This series of results formed an important supplement to the field survey and provided a reference for the risk assessment of geological hazards  in the a


We studied the magnetic properties of a glass-covered amorphous microwire that was stress-annealed at temperatures distributed along the microwire length. The Sixtus-Tonks, Kerr effect microscopy and magnetic impedance techniques have been applied. There was a transformation of the magnetic structure across the zones subjected to annealing at different temperatures. The annealing temperature distribution induces the graded magnetic anisotropy in the studied sample. The variety of the surface domain structures depending on the longitudinal location has been discovered. Spiral, circular, curved, elliptic and longitudinal domain structures coexist and replace each other in the process of magnetization reversal. The analysis of the obtained results was carried out based on the calculations of the magnetic structure, assuming the distribution of internal stresses. The Giant Magnetoimpedance (GMI) effect is determined by the remarkable change in the magnetic impedance in a wide range of frequencies when an external magnetic field is applied. This allows its sustainable and active use in magnetic, chemical and biochemical sensors. Magnetic wires, in which this effect is observed, have certain advantages that allow them to occupy a special place in the line of active elements ,2,3,4,5.Another application of microwires, which we took into account when choosing topics for this article, was their possible use in magnetic memory structures. The idea of controlled motion of domain walls is promising precisely for nano- and microwires, in which the observation of the formation and motion of domain walls is widely studied by a variety of experimental methods .As regards the background of the current research, a new direction of research related to the creation of microwire samples with longitudinally distributed magnetic properties should be noted here. As shown earlier, a characteristic feature of amorphous materials (and particularly of microwires) is that the anisotropy caused by stress annealing depends on the key parameter of the annealing temperature ,8,9. TheThe mentioned distributed magnetic anisotropy is achieved by a fairly simple method, which consists of annealing at a fixed applied mechanical stress and a variable annealing temperature. It is also important that such stress annealing is only partially reversible: subsequent annealing can restore only part of the anisotropy caused by stress annealing ,10. In aIn this article, we present our recent results on the study of the magnetic structure and magnetization reversal processes in magnetic microwires with distributed magnetic anisotropy caused in turn by distributed annealing temperature. The originality and novelty of this research is initially determined by our original idea to create samples with a continuous and uniform distribution of magnetic properties. The longitudinal distribution of the annealing temperature is a natural reason for the distribution of magnetic anisotropy, which in turn leads to a continuous distribution of magnetic properties along the length of the microwire. It is the continuity of changes in properties that is the object of our close scientific interest. Having carried out the first studies of such samples, in particular, the influence of graded magnetic anisotropy on domain wall propagation, we discovered the lack of knowledge about the features of the magnetic structure in this type of sample. Although we previously studied the magnetic structure in various annealed samples, it is the continuity of the change in the magnetic properties of the microwire in a wide temperature range from room temperature to 300 °C that is the decisive factor. Previously, we “locally” examined various annealed samples, and, as it turned out, thereby missed and did not fix various essential properties. Now this shortcoming has been eliminated. In particular, for the first time we observe the effect of the coexistence of various magnetic structures, replacing each other both in the process of magnetization reversal and when moving along the sample.64.04Fe5.71B15.88Si10.94Cr3.4Ni0.3  prepared by the Taylor-Ulitovsky technique  was created by attaching a mechanical load to one end of the microwire. The temperature, annT, inside the furnace was set to 300 °C. The annealing was carried out in air because insulating and continuous glass coatings protect microwires against oxidation. During the annealing process, a part of a sample was placed in a conventional furnace, while the rest of the sample remained outside the furnace. A tensile stress was applied during annealing. As a result, different parts of the sample were subjected to annealing at different temperatures . A tensiThe temperature was measured using a commercial  thermocouple inside the furnace, in the orifice (through which the microwire is introduced into the furnace) and near the furnace. As a result of the performed measurement, the distribution of the annealing temperature was obtained . LongituAX) co-directed with the microwire axis we have observed and fixed by microscope the surface magnetic structure transformation.The magneto-optical technique (Kerr effect) has been applied as a method to retrieve the information about the magnetization reversal (surface hysteresis loops) and to acquire the contrast images of the surface magnetic domain structures . ApplingTo measure the velocity of the volume domain wall we applied the modified method of Sixtus-Tonks ,12. The Furthermore, after obtaining the information of the shape of the EMF peaks for different locations in the long microwire sample, we use the Sixtus-Tonks technique for the estimation of the structure of the domain wall (DW) corresponding to the different locations in the studied microwire.max is the maximum applied magnetic field. Use of the mentioned technique allows us to measure the GMI effect in the extended frequency range up to 1 GHz.The impedance dependencies were evaluated using the specially designed micro-strip sample holder. The holder was placed inside a sufficiently long solenoid that creates a homogeneous magnetic field. The microwire impedance was determined from the reflection coefficient measured by the vector network analyzer ,14. We eThe GMI dependencies were measured for the locations in the studied microwire corresponding to the room temperature and to the annealing temperature of 300 °C. The small samples for GMI measurements were cut off from two opposite ends of the long sample, where the regions of interest to us were located. Other parts of the sample were not destroyed during this manipulation.The crystallization and Curie temperatures of the studied microwire, evaluated by the differential scanning calorimetry (DSC) method as described elsewhere , were abThus, the preliminary results obtained using the Sixtus-Tonks method were the reason for the using of the magneto-optic Kerr effect (MOKE) method for a more detailed study of the magnetization reversal and the rearrangement of the domain structure. The difference between these two methods is that the Sixtus-Tonks method provides information about the passage of a bulk domain wall along the whole cross section of the microwire, while the MOKE method shows the details of the domain structure and the magnetization reversal process in the microwire surface.A certain asymmetry of the hysteresis curves is observed at room temperature (RT) and 40 °C, while at temperatures of 60, 140 and 240 °C they are symmetrical enough. Being rectangular or nearly rectangular at temperatures close to room temperature, the MOKE hysteresis curves become progressively flatter as we move along the long sample towards the locations corresponding to higher temperatures of the annealing.A decisive addition, without which a detailed analysis could not be carried out, is that of the MOKE magnetic images of the structures obtained in the same places of the microwire where the MOKE hysteresis curves were measured . The men(a)Room temperature. Here we have determined that image 4b corresponds to the sharp jump in the MOKE hysteresis (right semi-loop of (b)Temperature of 40 °C. At this temperature, we also observe two different domain structures corresponding to two semi-loops b. The ve(c)For the location on the surface of the sample corresponding to the annealing temperature of 60 °C, the MOKE hysteresis loop is symmetrical c. This m(d)The magnetization reversal process in the location corresponding to the annealing temperature of 140 °C is also a combination of two types of domain structure g. We obs(e)Finally, in the location on the surface of the microwire corresponding to the annealing temperature of 240 °C, we found the domain structure which could be tentatively named “slow spiral structure,” which is characterized namely by the relatively low velocity of the motion along the microwire surface. Additional information for the analysis of the magnetic structures in different locations of the sample is provided by a cross-comparison of The wider and lower but still compact EMF peak b correspWe attribute the appearance of a double EMF peak observed at temperatures 60 °C and 140 °C to the coexistence of two types of domain structure in the process of magnetization reversal. The overall decrease in the amplitude of the EMF peak is associated with the turn of the magnetization in the magnetic domains in the direction of the circular direction and a decrease in the axial projection of the magnetization. For the temperature of 60 °C, one peak is associated with the formation of a multi-domain structure, and the second peak is associated with the rapid motion of a curved domain wall. For the temperature of 140 °C, the first peak is also related to the formation of the multi-domain structure, and the second peak reflects the motion of the circular domain wall separating the multi-domain and mono-domain magnetic state.Finally, for the location on the microwire surface corresponding to the annealing temperature of 240 °C, the results presented in Analyzing the obtained results, we identified several essential factors. For all the studied longitudinal locations on the surface of the microwire, the magnetization reversal process includes the simultaneous or successive coexistence of different types of domain magnetic structures. These structures can be divided into two main classes: those that change rapidly and those that change slowly. Behind this formal difference lies a more significant difference in structure, which we will discuss further.To explain the results obtained, we involved the results of theoretical simulations , as wellExperimental MOKE images presented in Theoretical calculations confirm this consideration. Applying the performed analysis to the experimental results presented in In the location of the microwire determined by the room temperature annealing, we find fast and slow elliptical structures. In the location where the annealing temperature was 40 °C, we find fast longitudinal and curved structures, which are special cases of the spiral domain structure. At an annealing temperature of 60 °C, one structure is replaced by another  during one half-cycle of magnetization reversal.The area of the microwire corresponding to the annealing temperature of 140 °C is characterized by the slow formation of an elliptical domain structure close to a circular one and the rapid movement of a single circular domain wall, which apparently has no correlation with the internal structure of the microwire.Finally, the domain structure observed in the longitudinal location associated with the annealing temperature of 240 °C is the spiral domain structure, with a small angle of inclination of the magnetization from the circular direction. Here we observe, namely, the motion of the whole domain structure, but not the domain wall. The domain structure moves by a formed front (marked by the red dashed line). According to our consideration, the low mobility of the motion of the front, separating the spiral domain structure and the mono-domain state, is determined by the strong correlation of this structure with the modified inner core inside the microwire.Taking into account the active use of microwires in magnetic sensors and for the sake of completeness of the analysis, we presented the results of GMI measurements carried out at two limit locations of the microwire. These locations correspond to room temperature and the annealing temperature of 300 °C . It is sThe difference in the value of the GMI ratio in the two studied locations is explained within the framework of the previously established relationship between the value of the GMI ratio and the angle of the helicity of the surface magnetic structure . As showAlong with sensors based on the GMI effect, microwires are active elements for sensors that use the magnetization reversal that occurs due to the fast motion of domain walls. In this regard, we would like to recall that, in addition to a significant GMI effect, a fast movement of a solitary domain wall is also observed in the location corresponding to RT. This rather unique property makes this area of the microwire the object of our further scientific attention in the frame of the sensor application of magnetic microwires.Our extensive research was aimed at the study of the magnetic properties of microwires with distributed magnetic properties. These distributed properties were determined by the distribution of magnetic anisotropy, which in turn was determined by the process of annealing at variable temperature. In our opinion, the most significant result obtained is that we managed to recognize a wide variety of the complex magnetic structures in various longitudinal locations on the surface of the microwire.We succeeded in determining the basic differences between these magnetic structures, which are manifested in the features of the magnetization reversal process. The complex use of the techniques allowed us to look at the problem from an unexpected angle and find those basic features of magnetic structures that were implemented in all the applied experimental techniques.To the analysis of the obtained results, we involved the results of well-proven theoretical calculations and experimental images of domain structures observed in magnetic semi-cylinders. As we were able to establish, the investigated domain structures can be divided into two classes, which are characterized by the velocity of their transformation and motion. We determined that the reason for this is the different degree of coupling of surface domains with the internal domain structure. The combination of helical, elliptical, curved, axial and circular domain structures changes smoothly as the point of observation moves along the microwire surface from room temperature to a maximum annealing temperature of 300 °C.Additionally, the studies of the domain wall velocity and GMI response give us the additional grounds to consider the studied microwire as a possible active element of magnetic sensors.


Antisense oligonucleotides (ASOs) are short single stranded synthetic RNA or DNA molecules, whereas double-stranded RNA nucleotide sequences are called small interfering RNA (siRNA). ASOs bind to complementary nucleic acid sequences impacting the associated functions of the targeted nucleic acids. They represent an emerging class of drugs that, through a revolutionary mechanism of action, aim to directly regulate disease-causing genes and their variants, providing an alternative tool to traditional “protein-specific” therapies. The majority of the ASOs are designed to treat orphan genetic disorders that in most of the cases are seriously disabling and still lacking an adequate therapy. In order to translate ASOs into clinical success, constant technological advances have been instrumental in overcoming several pharmacological, toxicological and formulation limitations. Accordingly, chemical structures have been recently implemented and new bio-conjugation and nanocarriers formulation strategies explored. The aim of this work is to offer an overview of the antisense technology with a comparative analysis of the oligonucleotides approved by the Food and Drug Administration (FDA) and the European Medicines Agency (EMA). Last century has witnessed the drug development process primarily based on the small molecules and antibodies . RecentlASOs are polymers consisting of 15–21 nucleotides . There aWhen single stranded, ASOs have to be stable and able to be selectively addressed to their target before the degradation due to circulating nucleases. On the other hand, the duplex structure of siRNAs makes them more stable than ASOs in the blood as well as within the cells. However, systemic delivery of double-stranded siRNA is more challenging as they have larger molecular weight and negative charge. Moreover, they have hydrophilic phosphates on their outside surface, while the aromatic nucleobases are located mainly inside the duplex, thus leading to scarce interactions with plasma membranes and quite rapid renal excretion .Once the ASOs has bound to the target RNA, a number of possible molecular mechanisms are triggered, which can be broadly classified as: (a) those promoting RNA cleavage through the recruitment of endogenous enzymes; (b) those that interfere with mRNA translation or maturation without promoting the degradation of the target . ASOs anOne of the common mechanisms for RNA target inactivation is the induction of RNase H, a family of endonucleases that selectively cleave the RNA strand of RNA/DNA hybrids . In the siRNAs act by using the RNA interference (RNAi) pathway, a natural cellular defence mechanism evolved to recognize and degrade pathogenic RNAs . For thisiRNAs enter the physiological process of RNAi by associating with TAR RNA-binding protein (TRBP) and the argonaut 2 enzyme (AGO2), composing the RNA-Induced Silencing Complex (RISC). Once the siRNA is part of RISC, one RNA strand is cleaved and released, whereas the remaining guide strand finds the complementary mRNA, forming an RNA-RNA hybrid that induces AGO2 to degrade the target RNA .The formation of RISC participates in cell-based defence, for example, against viruses. Despite that, siRNA-based drug applications are not limited to infective diseases, but they can be used for treating different types of cancers and various pathological conditions, including inflammatory disorders and neuropathies.When the guide or sense strand binds off-target transcripts, the RNAi-related mechanism of action of siRNA may cause unwanted activity. An important safety concern related to therapeutic siRNA mechanism of action is the possible non-specific suppression of non-target mRNAs by the passenger strand. Also ASOs can induce off-target effects by association with sequences having a high degree of homology or by interactions with proteins .Another mechanism used by ASOs for RNA inactivation is by preventing the interaction of mRNA with the ribosomes for reasons of steric hindrance. ASOs also interfere with mRNA maturation by constraining splicing or destabilizing nuclear pre-mRNA.Due to steric bulk, after pairing with the mRNA, an ASO can block the translation of the transcript with one of the following mechanisms: i) by hindering its contact with the ribosomal 40S subunit; ii) by avoiding the assemblage of the 40S/60S subunits; iii) by hampering the sliding of the ribosome along the transcript; iv) by impeding the interaction with sequences that are essential for the maturation of the transcript, such as the addition of a 7-methylguanosine cap at the 5′ end and polyadenylation at the 3′ end . Steric The goal of ASO therapy is to block the production of an abnormal form of protein or to restore the production of a protein that is lacking or not functioning . ASOs caMicroRNAs (miRNA) are small RNA molecules essential for gene regulation. They bind AGO proteins forming RISC, sharing the same proteins used by RNAi. Indeed, given that siRNA drugs compete with endogenous miRNAs to bind AGO2, it is important to optimize their dose to avoid RISC saturation and, in turn, inhibition of the RISC complex.A particular class of single-stranded DNA or RNA oligonucleotides is represented by aptamers. They have a three-dimensional structure that allows a highly specific interaction with proteins , this meHaving explored the diverse mechanisms by which ASOs exert their effects on target RNA, it is crucial to delve into their pharmacokinetics to better understand how these therapeutic molecules behave within the body. Understanding of pharmacokinetics is important in the drug development and safety field. This is particularly true for ASOs, where pharmacokinetics is intricately intertwined with the chemical properties of these therapeutic molecules. ASOs undergo various chemical modifications that impact their internucleotide links (first generation ASOs), the connecting rings between the ASO backbone and nucleobases (second generation ASOs), and the nucleobases themselves, which play a vital role in complementary base pairing with target RNA. These chemical alterations not only define the structural characteristics of ASOs but also govern their pharmacokinetic behavior, ultimately influencing their therapeutic effectiveness and safety profiles .The main route of administration for ASOs is the parenteral one (either intravenous or subcutaneous) . After sSubsequent to their release from the endolysosome, ASOs navigate to their designated site of action, endeavoring to evade degradation or re-exportation via exocytosis . FollowiThe use of chemical modifications has been crucial to increase ASO enzymatic stability, target recognition, binding efficiency, tissue distribution and, at the same time, to reduce their toxicity, leading to significant improved antisense drugs . The mosin vivo studies have demonstrated the necessity of high and repeated doses to achieve the therapeutic effect  and nucleic peptide acids (PNAs). PMOs are characterized by the substitution of ribose with morpholino and of the phosphorodextere bond with a phosphorodiamine bond. PNAs, on the other hand, are synthetic nucleic acid imitations containing neutral N-2-aminoethyl glycine units, with nucleobases linked by a flexible methyl carbonyl linker . Both clThe main positive effect of modifications applied to the heterocycle is the increase of the binding affinity to the target . One of Currently, modifications of the hydroxyl in the 2′ position of the sugar portion of deoxyribose in DNA and ribose in RNA, respectively, have made the greatest contribution in increasing the pharmacological properties of oligonucleotides by creating second-generation ASOs . The mosInstead Bridged nucleic acids (BNAs) contain a constrained bridge between 2′ oxygen and 4’ carbon of the ribose ring . These bin vivo, leading to improved symptoms in both severe and mild mouse models of spinal muscular atrophy (SMA)  .Advances in nanotechnology make it possible to overcome certain pharmacokinetic limitations of ASOs, including plasma and endosomal degradation, direct renal clearance and intracellular transmembrane delivery . The nanLipid-based delivery systems, such as lipoplexes and liposomes, derive from mixing polyanionic oligonucleotides with lipids. These formulations are able to mask the negative charge of the ASOs in order to reduce electrostatic repulsion with the cell surface . SpecifiPolymeric nanoparticles result from the non-covalent complexation between negatively charged ASOs and cationic polymers, including PEI (polyethylenimine), PBAE (poly-(beta-amino ester)) or poly-L-lysine . After eExosomes are biological nanoparticles whose use as delivery systems is current under study at a preclinical level. In particular, exosomes are extracellular bilayer vesicles, ranging in size from 30 to 100 nm, whose function is to facilitate intercellular communication through the transfer of nucleic acids, lipids and proteins . ExosomeSNA particles consist of a densely packed shell of nucleic acids, which are oriented around a hollow or solid core nanoparticle. SNA particles are currently undergoing clinical trials for the administration of ASOs to tumors (such as glioblastoma) or for their topical administration in the treatment of psoriasis .The potential delivery of ASOs to DNA and RNA can be enhanced by conjugation with ligands able to promote cellular uptake and active targeting by interaction with superficial receptors and, on the other hand, to reduce the direct renal clearance of ASOs by increasing their size . The mosin vivo activity of lipid-conjugated ASOs depends on their ability to bind plasma lipoproteins and thus exploit the endogenous system for lipid transport and uptake  to the siRNA passenger strand or single-stranded DNA ASOs . This liCell-penetrating peptides (CPPs) are short peptide sequences with positive charge that have been identified as carrying neutrally charged ASOs, with which they are linked, such as PMO and PNA, across cell membranes. CPPs can penetrate into the cell through endocytosis or by perturbing the plasma membrane lipid bilayer . The useEven though a number of technologies for hepatic delivery are available, extrahepatic targeting is still a challenge . CurrentThe most toxicologically characterised classes of ASOs are the first-generation ones (ASO-PS) and the second-generation ASOs with 2′-O-MOE sugar adjustments due to the larger number of drugs in clinical use. The toxicity of ASOs is dose-proportional and can be affected by both the oligonucleotide and the formulation .The toxicity of ASOs is commonly categorised into: hybridization-dependent or hybridization-independent way .Hybridization-dependent toxicity includes both excessive pharmacological effects that occur between the ASO and the target; and non-specific effects due to complementary or partial recognition of unwanted transcripts .in vitro effects of splice-modulating ASOs on 108 potential off-targets predicted on the basis of sequence complementarity, and identified 17 mis-splicing events for one of the ASOs tested  .iv. Thrombocytopenia - At the clinical level, thrombocytopenia is a side effect that may hamper the development of ASOs after the use of gapmer PS-2′-O-MOE . For exaThere are four subcategories of non-pharmacological toxicity:Proinflammatory and immunostimulatory effects are linked to the residual positive surface charge of some lipid nanoparticles used to promote siRNA biodistribution. Therefore, lipid nanoparticle formulations, as in the case of patisiran, require premedication with antihistamines, glucocorticoids and non-steroidal anti-inflammatory medicines to avoid unwanted effects .Encouraging results obtained mainly in preclinical studies suggest ASOs as interesting tools against oxidative stress. In C57BL/6J mice, an ASO was demonstrated to modulate the activity of a phosphatidylethanolamine N-methyltransferase (PEMT), that positively regulates mitochondrial ubiquinone (CoQ) content. Acute PEMT disruption mediated by ASOs was sufficient to increase mitochondrial CoQ and decrease superoxide, resulting in the preservation of insulin sensitivity and an improvement of glucose and insulin responses in high fat diet (HFD)-fed mice .Moreover, in HFD-fed mice, the treatment with generation 2.5 ASOs, which are highly potent and contain 2′–4′ constrained ethyl (cEt)-modifications that allow the binding to RNase H1.20, were used against the mammalian STE20-like protein kinase 3 (MST3) and resulted in protection against diet-induced oxidative stress at hepatic level, improving the full spectrum of HFD-induced nonalcoholic fatty liver disease, including suppressed liver steatosis, inflammation, fibrosis, and cellular damage . In partThe same authors had previously reported that the knockdown of MST3 in cultured human hepatocytes stimulates β-oxidation and triacylglycerol (TAG) and inhibits fatty acid influx and lipid synthesis, instead MST3 overexpression increases lipid accumulation in mouse and human liver cells. This resulted in an increase in density and size of intracellular lipid droplets, which is known to cause cellular dysfunction and contribute to the development of lipid-related metabolic disorders. The authors in fact reported that reducing or increasing MST3 abundance in human cultured hepatocytes leads to a suppression or an aggravation of oxidative stress, respectively .In a murine model of Alzheimer disease (AD), a phosphorothionated antisense against glycogen synthase kinase (GSK)-3β, GAO, lead to an improvement of learning and memory abilities coupled with a reduction of protein oxidation and lipid peroxidation markers . This dein vitro and showed protective effects against MPTP-induced dopaminergic terminal damage in vivo when injected into the striatum .Lastly, regarding miRNA inhibitors, an anti-miR-21 was demonstrated to be highly effective in murine models of Alport nephropathy: in particular, anti-miR-21 was able to reduce TGF-β-mediated stress response in glomeruli, to mitigate the expression of PPAR-α and its downstream fatty acid oxidation, to inhibit pro-inflammatory and profibrotic signals and to control the production of reactive oxygen species . Based oAs shown in The first oligonucleotide available for therapy was fomivirsen, a phosphorothioate ASO approved by the FDA and EMA respectively in 1998 and 1999 for the second-line treatment of cytomegalovirus (CMV) retinitis in patients with acquired immunodeficiency syndrome (AIDS) . FomivirThe first systemically approved ASO therapy was mipomersen, an antisense 20-mer phosphorothioate 2′-methoxy-ethoxy (MOE) gapmer, developed by Genzyme for adult patients with homozygous familial hypercholesterolaemia (HoFH) . HoFH isVolesorsen is a PS, 2′-O-MOE gapmer, approved by the EMA in May 2019, suitable for use in adults affected by familial chylomicronemia syndrome (FCS), a rare genetic disorder with high risk of pancreatitis, in case the response to diet and the therapeutic lowering of triglycerides is insufficient . VolanesDefibrotide is approved by EMA and by FDA, respectively in 2013 and in 2016, for the treatment of patients with hepatic veno-occlusive disease (VOD) that occurs after high dose chemotherapy and autologous bone marrow transplantation (HSCT) . Liver vInclisiran is a 2′F, 2′-O-Me, PS siRNA-GalNAc, approved by both the EMA and FDA on the basis of surrogate endpoints for the use in adults with primary hypercholesterolaemia or mixed dysplidaemia as a supplement to standard diet and therapy . These dInclisiran should be used in combination in patients who, with the maximum tolerated dose of statins, are unable to reach low-density lipoprotein cholesterol goals. Inclisiran, by activating AGO2, induces cleavage of the mRNA encoding for proprotein convertase subtilisin-kexin type 9 (PCSK9), an enzyme that negatively regulates the turnover and therefore LDL receptor (LDLR) levels on the hepatocyte membrane . This reGivosiran is a GalNAc-conjugated PS-siRNA used to treat acute hepatic porphyria (AHP) in adults and adolescents over 12 years of age . GivosirDeveloped by Alnylam Pharmaceuticals and approved in November 2020 by both the FDA and the EMA, lumasiran is a subcutaneously administered siRNA specifically to reduce hepatic overproduction of oxalate in patient with primary hyperoxaluria type 1 (PH1) . A high Nusinersen is an antisense 18-mer phosphorothioate 2′-methoxy-ethoxy (MOE) gapmer engineered to treat patients with Spinal muscular atrophy 5q (SMA), an autosomal recessive degenerative motor neuron disease . This diASO-based approaches aimed at exon 7 splicing correction  have empNusinersen increases the inclusion rate of exon 7 in SMN2 gene transcripts by binding to ISS-N1 and displacing splicing factors by steric hindrance . It is aOsredkar et al. recently have conducted a two-center study (in Slovenia and Czech Republic) which confirm the efficacy and safety of nusinersen after 14 months of treatment. The patient population under consideration included 61 subjects aged between 2 months and 19 years with a genetically confirmed diagnosis of SMA , a disease that primarily affects the nervous system causing vision loss, dysarthria, ataxia and dysphagia . It is cTransthyretin-mediated hereditary amyloidosis (hATTR) is caused by a mutation in the gene coding for transthyretin (TTR), a protein produced by the liver that transports thyroxine and retinol in the plasma and cerebrospinal fluid . The resInotersen is a 2′-O-MOE PS gapmer ASO that has been approved by both the EMA and FDA in 2018 for the treatment of hATTR . InotersDuchenne muscular dystrophy (DMD) is an autosomal recessive myophathy caused by mutations in the dystrophin gene. Patients usually experience premature death around 30/40 years of age due to respiratory or cardiac failure . Dystropin vivo studies (Golodirsen is a ASO with 25 monomers suitable for the treatment of DMD . Similar studies . Golodir studies . The appde novo dystrophin production in muscle (surrogate endpoint) and clinical improvements in time-based functional tests . It is administered intravenously once a week for 1 h at a dose of 80 mg/kg. Renal function monitoring is required for patients receiving viltolanersen. Viltolanersen received accelerated marketing approval from the FDA in August 2020 on the basis of a multicentre phase II clinical trial, which showed increases in al tests .Casimersen is an antisense phosphorodiamidate oligonucleotide designed for exon skipping in patients diagnosed with DMD confirmed by the susceptible exon 45 skip genotype . This drCasimersen is administered by intravenous infusion at the dose of 30 mg/kg once weekly .In February 2021, Casimersen received accelerated approval from the FDA based on the results of the ongoing, double-blind, placebo-controlled phase 3 study. Intermediate results showed a greater change in dystrophin expression from baseline in the casimersen treated group compared to placebo controls.The number of ASO molecules under investigation for the treatment of several diseases is considerable. The main candidates in phase III clinical trials are briefly described below in Major chemical and technological improvements have been made to diminish the toxicity and increase the efficacy of ASOs. This process has recently led to a great number of ASO-based medicines being used in clinical practice and to a renewed interest from big pharma, resulting in a multibillion-dollar market, comparable to that of small molecules. ASOs may be used in monotherapy or in combination with traditional therapy. They constitute a valid therapeutic alternative for diseases with no cure or effective therapies (orphan genetic diseases).Thanks to their properties, ASOs are able to regulate oxidative stress and inflammatory response that are common mechanisms altered in many disorders. Currently, ASOs are primarily intended for the treatment of neurological, neuromuscular and metabolic disorders. However, thanks to further technological innovations, ASOs are expected to be used as advanced therapeutics for many other disorders in the next future.


Open Science Graphs (OSGs) are scientific knowledge graphs representing different entities of the research lifecycle  and the relationships among them. They present a contextualized view of current research that supports discovery, re-use, reproducibility, monitoring, transparency and omni-comprehensive assessment. A Data Management Plan (DMP) contains information concerning both the research processes and the data collected, generated and/or re-used during a project’s lifetime. Automated solutions and workflows that connect DMPs with the actual data and other contextual information  are missing from the landscape. DMPs being submitted as deliverables also limit their findability. In an open and FAIR-enabling research ecosystem information linking between research processes and research outputs is essential. ARGOS tool for FAIR data management contributes to the OpenAIRE Research Graph (RG) and utilises its underlying services and trusted sources to progressively automate validation and automations of Research Data Management (RDM) practices.A comparative analysis was conducted between the data models of ARGOS and OpenAIRE Research Graph against the DMP Common Standard. Following this, we extended ARGOS with export format converters and semantic tagging, and the OpenAIRE RG with a DMP entity and semantics between existing entities and relationships. This enabled the integration of ARGOS machine actionable DMPs (ma-DMPs) to the OpenAIRE OSG, enriching and exposing DMPs as FAIR outputs.This paper, to our knowledge, is the first to introduce exposing ma-DMPs in OSGs and making the link between OSGs and DMPs, introducing the latter as entities in the research lifecycle. Further, it provides insight to ARGOS DMP service interoperability practices and integrations to populate the OpenAIRE Research Graph with DMP entities and relationships and strengthen both FAIRness of outputs as well as information exchange in a standard way. Funders’ and institutions’ mandates on open and FAIR 1 data mafree-form text documents that describe the processes, tools, and effort undertaken in data handling and management activities, including FAIR implementation, throughout a project lifecycle. The structure and expected content of the DMPs is provided in model templates prepared by the respective authorities who will receive researchers’ DMPs upon its submission. The most common requirements for DMPs are: (1) Data description and collection or re-use of existing data, (2) Documentation and data quality, (3) Storage and backup during the research process, (4) Legal and ethical requirements, codes of conduct, (5) Data sharing and long-term preservation, (6) Data management responsibilities and resources [DMPs have traditionally been esources . Today, esources .There have been early efforts to facilitate the DMP authoring process involving the development of software that guides researchers into the DMP contents and instructs them on how to answer the required questions to then download and share their work as .pdf files. DMPRoadmapRecently, Research and Innovation (R&I) ecosystems have started to realize Knowledge Graphs for their ability to present a contextualized view of current research that supports discovery, re-use, reproducibility, monitoring, transparency and omni-comprehensive assessment . In partAlthough important, DMPs are not yet integral parts of the ecosystem, and consequently, OSGs. Moreover, the adoption of ma-DMPs, i.e. machine actionable exports of DMPs that provide a structured way of organizing their content, is limited. Until today, DMPs are typically produced and submitted as deliverables outside of scientific workflows that support the publication of FAIR research outputs. Those practices pose several limitations on the DMP lifecycle, from the discovery and re-use of DMPs, to their enhancement with other information and further exploitation , 16.machine actionable’ DMPs (ma-DMPs) [In this respect, ARGOS (argos.openaire.eu)  was builma-DMPs) . OpenAIRma-DMPs) , 10 and lifecycle and enhance FAIRification of its outputs (DMPs are assigned DOIs and licenses). Finally, with DMPs being exposed in OpenAIRE (via Zenodo), the OpenAIRE RG became ARGOS main trusted source to: populate DMPs with standardized information, and create links with other outputs and activities .First, ARGOS applied the DMP Commons Standard  to achieThis paper, to our knowledge, is the first to explore the modelling, representation and linking of DMPs in OSGs. It explains the mapping activity that led to ARGOS DMP service integration with the OpenAIRE RG. The paper presents ARGOS as a case study for FAIR and ma-DMPs exposed in OSGs and highlights the new potential unlocked in the DMP landscape, from discovery to enrichment and exploitation.ARGOS DMP service is based on the OpenDMP open-source softwareARGOS is an instance of the OpenDMP open source softwareThe OpenDMP software features, among other things, a Template editor which is used by Administrators who define personalised structures and rules  for DMPs according to RDM policies. An example is the H2020 DMPs template  that canThe Template editor is a key enabler of compatibility with the RDA standard as it incorporates a mechanism that maps ARGOS OpenDMP fields contained in templates to template entities, including RDA dataset entities.OpenDMP has two more editors: the DMP Editor and the Dataset editor (embedded in the DMP editor) which are used by researchers who describe their datasets/outputs (e.g. how FAIR principles were followed throughout the RDM lifecycle) and project information (e.g. funding and beneficiaries) in DMPs. DMPs produced in OpenDMP enable values in DMPs to be fetched from external APIs, like OpenAIRE APIsAn essential element for achieving interoperability with standards and 3rd party systems, given the flexibility of the ARGOS underlying data model, which assumes almost unlimited user-driven structuring of dataset descriptions, is the concept of “semantic tags”. Semantic tags are tags that are attached to elements of a data set description template by their authors, so that, in due time, they utilized properly by an informed component that handles an intended action, be it an export or an interaction with an external system.A DMP’s lifecycle in ARGOS consists of a predefined number of internal states followed by versioning and publication workflows. A DMP is in draft state while being composed, and while draft it might be either validated  or non-validated, which is computed substate. Validated DMPs can advance to a finalized state, where editing is restricted, unless the DMP is reverted to draft again. The DMP may be published both internally, i.e. listing of published DMPs offered by the service, or externally.As shown in Figs. Once a DOI is issued from Zenodo, reverting a DMP to draft state is not possible and a new version must be created in order to add modifications. Following one cycle of publication, new versions of a DMP may be created and linked to their predecessor ones, while the latter continue to survive in the system for reference.The following table highlights how the ARGOS OpenDMP instance has enabled FAIRification of its outputs Fig. In summary, ARGOS DMPs are findable because they are minted DOIs directly from Zenodo, which in turn is registered in the re3data data repositories registryThe process of integrating ARGOS FAIR ma-DMPs with the OpenAIRE Research Graph concentrated on mapping the data models of OpenAIRE , OpenDMPThe aim was to identify what information captured in DMPs was already available in the graph, and vice versa, as well as how entities and properties are expressed in all settings, thus contributing to the mapping results. Since the DMP Common Standard is used to exchange DMPs in a standard way, it had a prominent place in this activity for providing the minimum set of DMP related properties that comprise a ma-DMP. Then, identified gaps and weaknesses were reported and prioritized for implementation as new entities and properties in both ARGOS and the OpenAIRE RG.The mappings were further analysed to report possible new relationships and properties that were not included in metadata records of DMPs found in OpenAIRE, but that are instead available in ma-DMPs like those produced and exposed by ARGOS.Of particular interest is the RDA HackathonThe comparative study  between The mapping highlighted the need for updates in ARGOS to be fully compliant to the RDA standard and strengthen its ma-DMP outputs. In Fig. Given that some researchers publish their DMPs in repositories, the OpenAIRE RG already included metadata about few DMPs made available by content providers as records compliant to the OpenAIRE guidelines , 10. MetAs it is shown in Fig. Those entities are:DMP_id), indicating the persistent identifier of the DMP record.Identifier  or be the compilation of free text statements provided in the DMP (non authoritative source). In Fig. Particularly for dataset metadata about ethics, security, quality, and preservation available in ma-DMPs, OpenAIRE could consider revisiting its guidelines , 10 so tBesides expanding information captured in the OpenAIRE RG, additional efforts are needed to create value out of this captured information. Below we describe what relation types are available in the OpenAIRE RG and what changed to accommodate links between ma-DMPs and other entities.The OpenAIRE Research Graph features a dedicated relation type between research products and project grants: “isProducedBy/produces” . This type of relationship can exist between projects and research products of any type , and is used by OpenAIRE to model the association between DMPs and project grants. Considering the importance assigned by funders to DMPs , and to support the development of added-value services on top of machine actionable DMPs, the OpenAIRE Research Graph will include a relationship with a dedicated semantics “hasDMP”/”hasProject”.Interestingly, the DMPs that are currently available in the OpenAIRE Research Graph do not include explicit links to the datasets they refer to. Clearly, such links may not exist when the first version of a DMP is published - because the datasets may not yet exist -, but the expectation is that the datasets are made available during the life-time of the project. Thanks to the different versions of the DMPs, therefore, it would be possible for OpenAIRE to add relationships with specific semantics between a DMP and the referred datasets. The relationships available in the OpenAIRE Research Graph, inspired from CERIF  and DataObservations made during the mapping activity of entities and properties between the three data models of OpenDMP, OpenAIRE RG and DMP Common Standard showed that when direct mapping couldn’t be fulfilled, information might still be able to be found in more abstract/ general fields of OpenAIRE or ARGOS though diverged in cardinality and / or data type; some information might still be covered by ARGOS DMP outputs as they enter the OpenAIRE RG or tweaked to accommodate the needs of ma-DMPs documentation or, rarely, they may be omitted in information exchange. OpenAIRE, also, highlighted the value added in contextualising ma-DMPs as they contain structured and specialised information, especially about datasets, which cannot be found in original / traditional DMP documents.Searching OpenAIRE for DMP outputs outside ARGOS, showed that DMPs are still typed as generic publications or reports, and not as data management plans. The introduction of a specific term for DMPs in global vocabularies, such as COAR’s, is expected to improve the current status, although the adoption of the new term may take some time to be widespread. The recent update of the Datacite metadata schemaFurthermore, in order to conform to the needs of ma-DMP fixed schema, without losing the versatility of its templating mechanism, ARGOS software follows an approach that engages an extensible mechanism for attaching export format converters (ma-DMP being one of them) and semantic tagging of template elements that can be used “at-will” by those converters. The ma-DMP converter makes use of its knowledge of the fixed part of ARGOS data model as well as attributes attached to various dataset description fields in order to pick the data required for a ma-DMP file.The flexibility of ARGOS machine actionable templating system combined with the integration of the publication mechanism of Zenodo and the interlinking with the OpenAIRE RG is crucial as it fosters change in the scene for DMPs content, distribution, exploitation, and reusability. According to the recent study of DMPs in Horizon 2020 , there aMoreover, it is foreseen that the OpenAIRE data model will be complemented with information that is currently not present, such as DMP cost or metadata. Cost information is not crucial to improve the level of FAIRness of DMPs or datasets, but it might enable analysis about data management costs useful to funders, organisations, and project administrators. Similarly, this kind of information is potentially useful to studies about responsible research and may be integrated to serve specific analysis or use cases related to RRI (Responsible Research & Innovation) monitoring.Finally, this case study explored information carried out in published, i.e. DOIed DMPs. An opportunity that ARGOS has already started to exploit with Zenodo is pre-filling of ma-DMPs with information from deposited metadata records. This automation highly contributes to the needed cultural shift that Open Science cultivates and incentivizes researchers to follow best practices while minimizing their compliance effort. Among the issues being examined in this implementation are capturing of the state of datasets that are pre-filled (e.g. reused dataset in a DMP) and adopting the pre-filling mechanism in other repository software, such as Dataverse. Further connections with data analysis PIDs are expected in the context of the Research Analysis Identifier System - RAISE project.The paper aimed to provide a case study of how machine actionable DMPs can be contextualised and exploited in an Open Science Graph, and specifically the OpenAIRE Research Graph. ARGOS, the OpenAIRE DMP service, contributed to this work by paving the way to exposing ma-DMPs in OpenAIRE.The paper explained how ARGOS is structured, how it applied the RDA standard to increase interoperability and machine-actionability of its outputs, how it integrated with other services to increase openness and FAIRness, and how it enriched the OpenAIRE RG. This is an ongoing effort between ARGOS and the OpenAIRE RG, supported thanks to the OpenAIRE API providing information about organizations, data sources and datasets, obtained by collecting metadata records from more than 12 K trusted scholarly communication sources .The results of the comparative analysis that was performed between the data models of ARGOS and OpenAIRE Research Graph against the DMP Common Standard extended ARGOS with export format converters and semantic tagging, and the OpenAIRE RG with a DMP entity and semantics between existing entities and relationships. This enabled the integration of ARGOS machine actionable DMPs (ma-DMPs) to the OpenAIRE OSG, enriching and exposing DMPs as FAIR outputs.Today, the work of this paper has shaped ARGOS in becoming the only DMP provider that completes the lifecycle by exposing its DMPs, including the ma-DMPs, directly to repositories and the only one to apply the whole set of the FAIR principles to its outputs so that DMPs are Findable, Accessible, Interoperable and Reusable.