With the gradual maturity of the terrestrial laser scanners (TLS) technology, it is widely used in the field of deformation monitoring due to its fast, automated, and non-contact data acquisition capabilities. The TLS technology has changed the traditional deformation monitoring mode which relies on single-point monitoring. This paper analyzes the application of TLS in deformation monitoring, especially in the field of ground surface, dam, tunnel, and tall constructions. We divide the methods for obtaining ground surface deformation into two categories: the method based on point cloud distance and the method based on displacement field. The advantages and disadvantages of the four methods  based on point cloud distance are analyzed and summarized. The deformation monitoring methods and precisions based on TLS for dams, tunnels, and tall constructions are summarized, as well as the various focuses of different monitoring objects. Additionally, their limitations and development directions in the corresponding fields are analyzed. The error sources of TLS point cloud data and error correction models are discussed. Finally, the limitations and future research directions of TLS in the field of deformation monitoring are presented in detail. With the development of human activities, there are more and more constructions. At the same time, disasters caused by construction deformation occur from time to time , causing great losses to human life and property safety. Therefore, the monitoring and analysis of the ground surface and constructions deformation has become increasingly important. It is necessary to regularly monitor the constructions to reasonably ensure and maintain their safety . ContinuThree-dimensional (3D) laser scanning is an emerging measurement technology, which is used to quickly and accurately capture points on the object surface with its advantages of fast speed, automation, non-contact, and high precision . Its appIn the past decade, a large number of studies have proved the feasibility of TLS in deformation monitoring, and deformation monitoring has been applied in surface, building, dam, tunnel, and other scenes . For theIn view of the keyword association in Under the influence of long-term geological processes and human factors, the original stress state of natural or artificial surface slopes will change, leading to stress redistribution and stress concentration, which will cause the surrounding rock and soil to experience different forms and even deformation, thus endangering the safety of nearby infrastructure and people. Therefore, it is important to monitor the safety and stability of slopes. The traditional monitoring methods include manual measurement, total station, GNSS, and close-range photogrammetry . AlthougThe surface slope deformation monitoring mainly calculates two types of quantities: (i) distance measurement between point clouds based on the variation difference with time; (ii) displacement field calculation based on the identification of corresponding elements in continuous point clouds.Methods for calculating point cloud distance have been developed for model-to-model (M2M), cloud-to-cloud (C2C) , cloud-tMany researchers use the method of point cloud distance measurement to obtain deformation information. In the study of the seasonal deformation on the Qinghai–Tibet plateau, TIN (triangulated irregular network) has the advantage of representing the digital elevation features by topographic feature points, and continuously calculating differences between multi-temporal TINs to obtain the elevation fluctuation of the Qinghai–Tibet plateau . The infIt is a common method to construct a 3D displacement field to represent the deformation direction of the surface area. The displacement field contains displacement vectors with corresponding directions, which represent the displacement change trend of the surface of the survey area. The displacement field operation often uses a sliding window to register the regional point cloud to obtain the displacement vector. In other words, the difference in the method based on displacement field is the different way of point cloud registration.The cross-correlation-based particle image velocimetry (PIV) method was applied to derive a surface deformation field . Point cThe two methods based on point cloud distance and displacement field are the most commonly used methods in surface slope monitoring. The method based on point cloud distance is used more frequently than the method based on displacement field. Except for C2C and M3C2 that directly compare point clouds, other methods all need to establish different types of models. This requires selecting appropriate models for different monitoring scenarios. Simple triangular meshes can be established for those with small fluctuations, while polynomial fitting is often used for those with surface irregularity. At present, there is a lack of unified evaluation of model accuracy, in addition, there is not much research that gives appropriate type correspondence for selecting fitting methods. The displacement field technology does not need to face this problem. The core of the displacement field technology is the matching of point cloud features. Through sliding windows, roughness, and other point cloud features, it is possible to match the differences in features of point clouds at different periods. The key is the method of feature extraction, which is more difficult than point cloud distance measurement. From the above analysis, it can be seen that the average accuracy of TLS in surface deformation monitoring is basically at the centimeter level. This is because the surface deformation area is large, and the point cloud data quality decreases with the increase of distance when TLS scans over long distances.As an important part of river flood control projects, dams realize the reasonable regulation and optimal allocation of water resources through water storage, which is very important to produce electricity, water supply, and irrigation. They play an important role in the lifeblood of society and economy . HoweverA reconstruction algorithm based on spherical projection was proposed , which uIn dam deformation monitoring, deformation information should be extracted from the model rather than the point cloud. First, a complete dam model was constructed from the point cloud, including triangulation model, contour model, and function fitting model. Since the point cloud data will inevitably encounter the situation of leakage holes and noise points, it was necessary to interpolate the point cloud data during modeling, which reduced the influence of point cloud roughness. At the same time, the question of whether the point cloud obtained by interpolation was consistent with the actual surface condition deserved consideration. As a means to obtain deformation information, the accuracy of the model affected the accuracy of the final deformation monitoring results directly. The current stage is to explore the improvement of model accuracy, from TIN, the linear contour model, and then to the function fitting model. The high accuracy of the dam surface model is the key goal we pursued. Due to the dam being monitored over a smaller area, the accuracy in deformation monitoring of the dam was at millimeter level.With the development and utilization of urban underground space, tunnels are becoming more and more common. Laser scanning technology for deformation monitoring in the tunnel engineering application has caused some concern , especiaAiming at the problem of how to effectively obtain 3D models and accurately extract the feature information of large-scale composite structures such as tunnels, a surface-based nondestructive measurement method was proposed and developed, and its accuracy falls within the millimeter range . The metIn recent years, the application of TLS technology in engineering surveys has become a technology for large-scale applications in the tunnel engineering environment, due to its advantages of non-contact, fast speed, high precision, and large-scale data acquisition. In order to obtain satisfactory scanning data, this section briefly introduces the application in tunnel monitoring. In tunnel deformation monitoring work, the first thing to do is to extract the central axis, and then slice the tunnel point cloud to fit the cross-sectional analysis. In tunnel deformation analysis, the accurate extraction of the axis is particularly important, and how to fit the central axis accurately and quickly is the key research direction. From the above research, it can be seen that the accuracy of TLS in tunnel axis extraction and section fitting is very high, at the millimeter level. In addition, the geometric deformation of the tunnel is monitored during excavation, and the accuracy and reliability of the 3D reconstruction of the tunnel scene are also to be studied. The real scene in the tunnel is 3D, and the regular monitoring modeling comparison is to determine the deformation trend of the tunnel. Moreover, the recognition of tunnel cracks or leakage areas should also rely on deep learning and image recognition algorithms, after all, image recognition algorithms are more mature, and point cloud intensity information can also be used to monitor water leakage and cracks. TLS has not yet become a common tool for tunnel measurements, but it still has great potential for exploitation.Some scholars have used TLS to monitor the deformation of some tall constructions. The most common characteristics of these constructions is that they are very tall. Most tall constructions have complex multi-story structures, such as cooling towers, wind turbine towers, and chimneys. The cross-sections of these constructions are usually circular or elliptical. The long-term influence of external loads on these structures causes different degrees of deformation of the structure. This not only affects the stability of the construction itself, but also threatens the security of people’s lives and property. Monitoring the deformation of tall constructions can do a good job of risk transfer and early warning before the risk occurs. Therefore, it is important to monitor the deformation of tall constructions.x, y coordinates and average adjustment errors of the cylindrical axes could be calculated. By comparing the x and y coordinates of the higher layer with those of the lowest layer, the deviation of the chimney axis from the vertical direction was calculated [Cross-sectional method: surveyors generally use the line scanning mode of observation to fit circles at different heights, and to fit the center coordinates of circles of thin point cloud slices at multiple levels. By comparing the center coordinates of the upper levels with those of the lowest levels, the 2D displacement deviations of the tower moving in the axial and lateral directions can be determined. Some researchers used TLS to conduct line scans of cooling towers, wind turbine towers, and chimneys to analyze axial deformation monitoring , and uselculated . For thelculated .y axis of the building’s central axis and the horizontal x axis, and determining whether the building is inclined due to ground subsidence by the degree of the angle. An angle of 90° indicates no inclination and less than 90° indicates that the building has been inclined.Surface parametric modeling: Gauss–Helmet nonlinear model was used to estimate the surface parameters of a cylinder for 3D parametric modeling . This meIn addition, some scholars have used not only TLS, but also combined it with other technologies to monitor the deformation of tall constructions. Based on the TLS technology, the video measurement was also used , with thThe TLS technology with rapid survey, large number of points, and high recognition accuracy is very effective in surveying the geometry of objects that change rapidly over time. However, its limitation is the repeatability, which requires the identification of new points on the surface each time. The advantage of the cross-section method is the simplicity of the algorithm and its wide application. However, its limitation is that it is not global monitoring, but uses the coordinate difference of the center of the fitting circle of the point cloud cross-section at different heights to determine the axial deviation of the object, and does not use all the point cloud data. The advantage of the surface parametric modeling method is that it directly obtains the inclination angle of the structure axis, avoids the blind spots between the cross sections, and provides a more advanced analysis of the structural deformation compared with the modeling surface. Both methods are more effective when the shape of the monitored object is regular and can be expressed by functions. In addition, other means  can be combined to achieve more comprehensive monitoring when monitoring the deformation of tall constructions.As numerous studies and applications have been conducted in the field of deformation monitoring, data acquisition is the premise and basis of deformation analysis. It includes inspection of instruments, field survey, control network layout, and scanning plan formulation. It is necessary to check whether the scanner can work normally and whether its accuracy can meet the engineering requirements; then, to go to the site to survey the terrain. For different terrain conditions, the subsequent deployment of the control network and determination of the scanning position of the scanner should be considered in advance. After investigating the terrain, the control network can be established, which is not necessary but recommended. Control points can improve the registration accuracy, which is conducive to transform the local coordinate system of the point cloud into the unified coordinate system. Finally, the scanning plan is formulated, and operators acquire the point cloud data according to the scanning plan.In the application of TLS technology, data preprocessing is also an extremely important aspect . The firIn deformation monitoring projects, multi-station splicing of point clouds in the same period is usually done through white balls or checkerboard markers ,21, whicIn the process of laser scanning, point clouds of unwanted objects such as vegetation and electric wires will inevitably be collected and treated as noise points, which will affect deformation monitoring and analysis ,79. CorrAs a common step in all deformation monitoring projects, the registration and filtering of point clouds are sufficient to prove their importance in deformation monitoring studies. The registration speed and accuracy are important indicators for evaluating the registration method, and they are also the directions of future research. Faster point cloud registration can reduce the preprocessing time of point clouds, and higher registration accuracy can build a model that is more consistent with the surface and obtain higher shape variables. Point cloud filtering is required to improve the degree of automation and the accuracy. Nowadays, some projects still use the manual method to remove noise, which undoubtedly increases the time of data processing. How to completely and accurately remove noise points to ensure that the real surface data is not removed is a key issue. Currently, many algorithms can meet the basic requirements, but the presence of noise point outliers is fatal for more precise deformation monitoring projects. It is also a factor restricting the application of TLS in high-precision deformation monitoring. In the future, the filtering algorithm should be developed in a fast and accurate way to promote the application of TLS technology.Millimeter accuracy is often required in the deformation monitoring industry. Therefore, data quality is one of the most important factors for the effective use of TLS in the deformation monitoring industry. As with traditional techniques, TLS is subject to different sources of uncertainty during surveying. In this case, it is crucial to identify potential sources of errors which affect the data quality and assess their impact on the results.When using TLS, the accuracy of point cloud data can be affected by instrument mechanisms, atmospheric conditions, object surface properties, and scan geometry ,83, and In the future, the impact of various errors on the accuracy of point cloud data should be solved from following two aspects. Firstly, research and development of the TLS equipment should be conducted to improve the scanning distance of the TLS, reduce the scanning limitation caused by the incidence angle, improve the scanning distance and accuracy of the TLS, and select a laser that is less affected by environmental factors as the light source. Secondly, the error correction models should be studied, and the algorithm model should be applied to improve the point cloud data quality. The error models of atmospheric refraction, point cloud reflection, and other influencing factors should be established. The correction model of temperature and humidity should be improved through experiments and analysis.With the development of TLS technology, the proportion of its application in deformation monitoring has gradually increased. In this paper, we reviewed the latest development of deformation monitoring based on TLS, especially the deformation monitoring methods used in the fields of surface, tunnel, dam, and tall constructions. We summarized the differences in the methods and the focus of extracting deformation information in different domains. In ground surface and slope monitoring, there are two methods based on point cloud distance measurement and characteristic displacement. Most dam monitoring methods are based on the difference of models. In tunnel monitoring, we should pay attention to the extraction of the central axis, segment dislocation, and leakage cracks. In tall constructions monitoring, the cross-sectional method and surface parametric modeling are used to analyze their tilt and offset. Then, we presented the main steps of point cloud preprocessing, including registration and filtering, and analyzed the factors that affect the quality of point cloud data, as well as the factors limiting the development of TLS were discussed. Finally, we can see from the above research that the accuracy of the TLS is unfixed for different monitoring objects. When monitoring the surface displacement or settlement, the observation range is often large and some places are far away from the scanner, resulting in incomplete point cloud data, which reduces the final overall accuracy. This is the limitation of the application of TLS. However, when monitoring in a small range, objects can be scanned in detail, and the accuracy can mostly be within the millimeter level, so TLS can play an efficient role in short distance and small range observation.In order to promote the development and application of TLS in deformation monitoring, this paper analyzes and proposes the following future research directions:(1) Improve the accuracy of the data acquired by the instrument. From the above analysis, we can know that TLS acquires very detailed data in short distance and small ranges, but with the increase in the observation distance, data quality becomes more and more poor, which limits the widespread application. Instrument manufacturers should develop more precise scanners suitable for remote measurement to adapt to deformation monitoring in different scenarios.(2) Improve the data processing capability. Although a lot of research has been conducted on data processing, there is still a considerable gap between the existing technology and its application requirements. There is great potential in improving the processing efficiency, robustness, and automation of algorithms. In addition, the developed algorithms should be oriented to practical applications and have universal applicability in the field of deformation monitoring. Finally, a standard system should be established to evaluate the performance of the algorithms in order to select appropriate methods for different applications.(3) Establish a high-precision point cloud model. When TLS is used for deformation monitoring, the scanning point cloud modeling is used to obtain the overall information of the object surface. When the point cloud data is lost , the sel(4) Artificial intelligence (AI) is adopted. At present, AI has been widely used in all walks of life. As a branch of computer science, it has great advantages in processing and analyzing a large amount of data. There is no doubt that AI will become one of the main trends in the field of deformation monitoring in the future. In recent years, various AI techniques, especially deep learning, have been found in previous studies proving its great potential in object detection and quality assessment ,99,100. (5) Use BIM (building information modeling) technology. It is also a trend of point cloud application. Through the construction of building model information, the characteristics of objects and their surrounding environment can be intuitively seen. To achieve an integrated project, i.e., integrate and manage each project phase and management  a BIM approach becomes indispensable . In addi(6) Combined with VR/AR  technology. In the current workflow of using point cloud data for architectural applications, the acquisition of point cloud data is usually separated from the visualization and processing of point cloud data. Due to process fragmentation, engineers are unable to obtain meaningful construction activity information from point cloud data in time. Its delay prevents the timely identification and processing of on-site problems, especially those related to geometric quality inspection and construction progress tracking. Due to this limitation, future research needs to develop methods for real-time visualization and processing of point cloud data for practical applications. One possible solution is to combine point cloud acquisition and processing tools with VR/AR technology . Taking


Alpheus bellulus in a tank using an X-ray computed tomography (CT) scanner. CT scanning was performed at 10–30 min intervals immediately after the start of burrow construction. The three-dimensional morphology  of burrows at each observation time was imaged and measured. In addition, the rate of increase of each parameter was calculated. Surface area, volume, length, and depth rapidly increased immediately after the start of the experiment in all burrows. Subsequently, there was a reduction in the rate of increase at 40 min after the start of excavation for burrow depth, at 75 min for length, and at 90 min for surface area and volume. Although there were large differences in burrow diameter among the burrows immediately after the start of the experiment, all burrows reached nearly identical diameters after 90 min. Changes in burrow morphology were not observed in most of the burrows more than 210 min after the start of the experiment, meaning that A. bellulus can create burrows that are sufficient for survival within this time period. The use of CT scans in this study clarified the developmental process of the three-dimensional structure of A. bellulus burrows and is applicable to various burrow-producing organisms. Our results provide new insights into the development of burrow structures.The burrow morphology of endobenthic organisms reflects their subsurface ecology. In this study, we observed the three-dimensional development of burrows produced by the tiger pistol shrimp  Therefore, much of their subsurface ecology remains unknown.Burrows in marine environments have existed throughout the world since the Cambrian explosion when benthic organisms started to make a burrow acting as a refuge from predators , and havThe burrows of benthic organisms are generally characteristic, and can be distinguished by taxon or feeding ecology . AlthougAlpheus bellulus  in experimental aquaria  individuals in the laboratory for this study. These individuals were sourced from an aquarium shop in Tsuchiura, Ibaraki Prefecture, Japan. This species is widely distributed in the Indo-Western Pacific, and its congeners are common in tropical and temperate shallow marine settings throughout the world. Of the more than 300 Alpheus species described so far [Alpheus bellulus has been identified in Japan from the Izu Peninsula [A. bellulus burrows reach 30–70 cm in length and 70 cm in depth [Amblyeleotris japonica, Amblyeleotris diagonalis [Tomiyamicthys oni [Stonogobiops pentafaciata [We observed five d so far , about 6eninsula , Kii Peneninsula , Okinawaeninsula , and Yaeeninsula . The shrin depth . Burrowsagonalis , Tomiyamthys oni , and Stoafaciata .A. djiboutensis has been reported to show negative phototaxis [The five study animals were identified as individual A (IA) through E (IE). Their carapace lengths were 13.6 mm, 19.4 mm, 13.3 mm, 17.4 mm, and 15.8 mm, respectively. Prior to experiments, all shrimps were maintained in the laboratory under the same constant environmental conditions as those of the experiment for over 10 days to ensure that any endogenous or physiological rhythms were abolished. IA and IB were first housed in their experimental tanks on 22 July 2020, and IC, ID, and IE were first housed on 17 September 2020. Each shrimp was placed in a separate plastic tank . No negative effects of the repeated X-ray scanning on shrimp burrowing activity were observed during the experiment; burrowing activity by the shrimp after the end of the experiment appeared to be the same as the activity observed prior to the experiment.After acclimatization to constant environmental conditions for 10 days or more, tanks with active burrows were subjected to X-ray CT scanning  at power settings of 80 kV and 250 mA and a slice thickness of 0.625 mm. The pixel area was 0.59 mmp-value did not exceed 0.05.The Amira 2019.2 software platform with X Image PAQ Extension  was used to analyze three-dimensional images obtained by the X-ray CT scans. The cross-sectional  shape of the burrows in each X-ray image were traced to obtain three-dimensional images of the burrows at each scanning timepoint . The surA. bellulus shrimp  and volume . However, there was almost no correlation between carapace length and burrow surface area , volume , depth , length , or diameter .The relationship between carapace length and the maximum value of each burrow parameter was as follows: There was a positive, though not statistically significant, correlation between carapace length and the rates of increase of burrow surface area  might have been sufficient to see an effect of carapace length on burrow morphology because range width of carapace lengths in Henmi et al [Henmi et al.  made resmi et al  was simiOne possible factor is the limited time and space provided for burrow development in our study. Henmi et al.  measuredAnother possible factor is the size of the shading plate used in our experiment. All shrimp in our study placed their burrow openings near the coral rock and constructed their burrows exclusively under the shading plate. Considering the diagonal length of the plate (about 330 mm) and the diameter of the coral rock (about 30 mm), the maximum horizontal length of any burrow that satisfies these conditions would be about 360 mm . This buA. brevicristatus [A. brevicristatus have a branch-like structure in which the burrow diameter matches the size of the body [A. bellulus burrows observed in our study did not have a branch-like structure, and the bulk of the burrow consisted of the chamber dug under the shading plate. The diameter of this chamber did not appear to be correlated to carapace length. In addition, the lack of correlation can be attributed to the small sample size (n = 5).Although we did not find any correlation between carapace length and burrow diameter in our study, significant positive correlations have been reported for the congeneric ristatus . This isthe body . HoweverLasius flavus and found that exogenous environmental features such as sediment boundaries affect nest morphology. The advantage of this method is that it allows researchers to nondestructively observe temporal changes in nest internal structures in three dimensions [Minter et al.  used micmensions . HoweverL. flavus was not affected by exposure to X-ray CT scanning, and in the present study, the A. bellulus individuals used in our research were still constructing burrows more than a year after our X-ray CT experiments . This suggests that the levels of radiation used in our experiment will not affect the survival and behavior of other large crustaceans, including other Alpheus shrimps, but more data is needed to confirm this. CT scanning of burrows will likely prove to be a highly useful and versatile method for quickly obtaining quantitative four-dimensional data on burrow morphology.Although repeated exposure to X-ray radiation could conceivably alter the behavior of experimental animals, this is unlikely to have affected our results. Minter et al.  reported


Aberrant protein phase transitions and aggregation trigger various neurodegenerative diseases, including ALS, FTD, Alzheimer’s disease, and Huntington’s disease. Dynamic exchange of proteins and RNA between the nucleus and cytoplasm is essential for cellular functioning, which is greatly affected by the mislocalization, phase separation, and aggregation of different RNA-binding proteins (RBPs), such as FUS, TDP-43, and hnRNPA1/A2 in their respective neurodegenerative diseases. Disease-linked abnormal accumulation of these RBPs impairs the nucleocytoplasmic transport (NCT) by disrupting the nuclear envelope architecture and mislocalizing essential NCT factors, such as nucleoporins, importins, exportins, and the Ran protein and its regulatory components. Nuclear import receptors (NIRs) have the potential to restore abnormal mislocalization of RBPs by regulating the import/export pathway. NIRs can also potentially prevent or reverse the phase separation and fibrillization of RBPs. Overall, this review focuses on the role of nuclear import receptors in regulating phase transitions in neurodegenerative diseases.RNA-binding proteins (RBPs) with a low-complexity prion-like domain (PLD) can undergo aberrant phase transitions and have been implicated in neurodegenerative diseases such as ALS and FTD. Several nuclear RBPs mislocalize to cytoplasmic inclusions in disease conditions. Impairment in nucleocytoplasmic transport is another major event observed in ageing and in neurodegenerative disorders. Nuclear import receptors (NIRs) regulate the nucleocytoplasmic transport of different RBPs bearing a nuclear localization signal by restoring their nuclear localization. NIRs can also specifically dissolve or prevent the aggregation and liquid–liquid phase separation of wild-type or disease-linked mutant RBPs, due to their chaperoning activity. This review focuses on the LLPS of intrinsically disordered proteins and the role of NIRs in regulating LLPS in neurodegeneration. This review also discusses the implication of NIRs as therapeutic agents in neurogenerative diseases. Liquid–liquid phase separation (LLPS) is an indispensable and pervasive phenomenon underlying the formation of membraneless organelles and biological condensates throughout the cells ,2. A groNucleocytoplasmic transport is a bidirectional communication between nucleus and cytoplasm required for the transport of different proteins, RNAs, and ribonucleoproteins . A complNups are categorized based on their location within the NPC A 28]. T. T28]. TNuclear transport receptors belong to a protein family called karyopherins (Kaps) that includes importins and exportins . ImportiThe Ran protein, which is a member of the Ras superfamily of small GTPases, plays a significant role in nucleocytoplasmic transport. Ran is highly ubiquitous and found in two forms in mammals: Ran guanosine diphosphate (RanGDP) and Ran guanosine triphosphate (RanGTP). GTP hydrolysis by Ran to form RanGDP from RanGTP is responsible for a large portion of the energy required for nucleocytoplasmic transport. Moreover, Kaps associate with RanGTP or RanGDP to regulate the directionality of nucleocytoplasmic transport ,41,42. SA nuclear localization signal (NLS) is a short peptide sequence of positively charged amino acids, including lysine and arginine, that acts as a signal peptide to mediate the transport of proteins from cytoplasm to the nucleus ,47. NIRsSignal-umpired nuclear localization is mediated by NIRs that are members of the karyopherin family of proteins, including importin αs (or Kapαs) and importin βs (or Kapβs). Importin αs are adapter proteins that connect cNLS-containing proteins with importin β for nuclear import . ImportiImportin βs are another family of nuclear transport receptors that include importins, exportins, and bidirectional receptors ,88. TherRan gradient across the nuclear membrane regulates the directionality of nuclear transport ,41,42. FA nuclear export signal (NES) is a short signal peptide of four to five hydrophobic amino acids in the proteins that directs the protein out of the nucleus and is recognized by exportins. An NES affects several vital functions of the cell, including transcription and nuclear export  A typicaThe chromosome region maintenance 1 protein (CRM1) or Exportin-1 (Xpo1) is the leading karyopherin that helps with the export of most protein cargos that have a leucine-rich NES  96]. CR. CR96]. The roles of CRM1 in the nuclear export of TDP-43 ,107 and Our understanding of membraneless organelles has increased with developments in the field of liquid–liquid phase separation research. LLPS is involved in the formation of many membraneless organelles, such as P bodies, stress granules, nuclear Cajal bodies, nucleolar body, nuclear speckles, and carboxysomes ,111,112.NIR cargos, such as FUS, TDP-43, and hnRNPA1, which are IDR-containing RBPs, can undergo liquid–liquid phase separation and are involved in different neurodegenerative diseases ,117,118.Fused-in sarcoma (FUS) is a RNA-binding protein that plays a role in RNA transcription, splicing, transport, and translation . FUS plaTDP-43 has been identified as one of the major disease proteins aggregating in ALS and FTD patients . It is aD262V is more associated with stress granule assembly [D262V mutant can promote its amyloid-like aggregation [hnRNPA1 is a 320 amino acid long protein and a member of heterogeneous nuclear ribonuclear protein (hnRNP) family . It compassembly . The LLPregation .A shared pathological feature of multiple neurodegenerative diseases is the presence of large intracellular or extracellular inclusions in the brain of diseased patients that cannot be degraded by normal mechanisms. Protein aggregates could induce nucleocytoplasmic defects by sequestering components of the NPC or directly blocking the NPC, leading to protein mislocalization and aggregation ,148,149.ALS is a late-onset, relentlessly progressive, prototypic age-dependent neurodegenerative disorder caused by the loss of motor neurons, leading to paralysis and death ,163. TheFTLD is a progressive neurodegenerative disease associated with neuronal degeneration in the frontal and temporal lobes, with neuronal intranuclear and cytoplasmic inclusions ,168. FTLBoth diseases are heterogeneous and share some clinical, neuropathological, and genetic features . In addiDifferent reports indicated that impairment in the NCT is one the major mechanisms of ALS and FTLD pathology. Cytoplasmic TDP-43 aggregates were found to be accumulated with many Nups and transport factors, suggesting their role in disrupting the NCT ,199. TheThe GGGGCC (G4C2) hexanucleotide repeat expansion (HRE) in C9orf72 is the most common genetic cause of fALS and fFTD ,203. G4CFurther, mutations in other proteins implicated in ALS, such as senataxin and Vesicle-associated membrane protein-associated protein B/C can cause impairment in the NCT ,154,155.Alzheimer’s disease is a chronic neurodegenerative disease with well-defined pathological mechanisms. It contributes to 60% to 80% of dementia cases . NeuritiRecently, disruption of neuronal NCT was observed as one of the major pathological features involved in AD . Using aThe mechanism of NCT impairment in AD has been studied in the context of Aβ and tau aggregates. A study demonstrated that oligomeric Aβ species can reduce the Ran expression level and disrupt the nucleocytoplasmic transport in AD models . AdditioHuntington’s disease (HD) is a rare, inherited, and progressive neurodegenerative disease caused by CAG-repeat expansion in exon 1 of the huntingtin gene, leading to the expression of mutant huntingtin (Htt) protein with expanded polyglutamine (polyQ) repeat ,239. HttThe prevalence of most neurodegenerative diseases, including AD, PD, and ALS, rises with age. Age-related neurodegenerative disorders have few or no effective therapies, and they tend to proceed in an irreversible way, resulting in high economical and human consequences . Normal Some differentiated cells, such as muscle fibers and neurons, have long-lived scaffold nucleoporin proteins, such as Nup93 and Nup107 . These lThe aforementioned neurodegenerative diseases target different regions of the brain, but they all selectively target a subpopulation of neurons, leading to the progressive failure of defined nervous system regions. Selective neuronal vulnerability is a shared property of these neurodegenerative diseases, but the basis of such selective neuronal vulnerability has remained elusive. Interestingly, different neuronal cells showed different susceptibility to nucleocytoplasmic transport defect . This diRegardless of the significant evidence of NCT difficulties in ALS, FTD, HD, and AD, there is currently no treatment option for neurodegenerative disorders that targets nuclear transport impairments. Finding new approaches to repair NCT disruption in neurodegenerative proteinopathies is a tantalizing new potential approach for avoiding neuronal death in these disorders, but it is also a daunting task. The tremendous molecular and structural complexity of the NPC, as well as the relevance of NCT for practically all cellular activities, present major obstacles in designing treatment techniques. NPC disturbances frequently result in an imbalance in the nucleocytoplasmic gradient of NTFs, transcription factors, nuclear proteins, and RNA, which can be partially reversed by boosting or inhibiting nuclear import or export ,266. CRMNIRs employ nuclear-localization signal of polypeptide cargo in the cytoplasm for their transport to the nucleoplasm. Through binding to NLS, NIRs have the potential to inhibit and reverse the aberrant phase transitions of several intrinsically disordered RNA-binding proteins with prion-like domains, such as FUS, TDP-43, hnRNPA1, hnRNPA2, EWSR1, and TAF15, as well as their disease-linked mutant variants . Higher In addition to Kapβ2 , a few other nuclear import receptors, including transportin-3, importin β, importin 7, or the importin β/7 heterodimers, directly bind to FUS via its RG/RGG motifs . In addiRAN translation of the C9orf72 HRE produces arginine-rich DPRs (R-DPRs)  and poly (PR)), which are the most toxic of the five DPRs generated in neurons . These RThe chaperone and disaggregase activities of nuclear import receptor family members have become an attractive target for therapeutic development in conditions involving the protein aggregates. Screening for drug-like molecules that improve NIRs’ ability to prevent or reverse RBP fibrillization will be very interesting . DiffereMoreover, a chaperone’s activity in cell can be boosted by small molecules that can boost their expression level, which can be screened using cell-based models . In addiMany proteins rely on their intracellular localization to function properly. Therefore, NCT is critical for a wide range of physiological and pathological processes. Defects in NCT have been associated with several neurodegenerative disorders characterized by protein mislocalization and the formation of toxic protein aggregates resulting from aberrant protein phase transition. These aggregates can trigger more alterations in the NCT machinery, either by causing morphological changes in the nuclear pore complex and nuclear membrane or by the mislocalization of NCT components, including Nups, importins (NIRs), exportins, and other regulatory factors. Although NCT impairments have been linked to several neurodegenerative disorders and ageing, no therapeutic strategies are available. Nuclear import receptors can act as a chaperone and have the potential to inhibit or reverse the aberrant phase separation and toxic aggregation of different RBPs. They can also effectively restore the nuclear localization of mislocalized RBPs to their normal function. Thus, potentiating NIRs’ activity using small molecules or genetic approaches to prevent the mislocalization and aberrant phase transition of RBPs involved in disease pathology may be a promising technique.


The progress of science increasingly relies on machine learning (ML) and machines work alongside humans in various domains of science. This study investigates the team structure of ML-related projects and analyzes the contribution of ML to scientific knowledge production under different team structure, drawing on bibliometric analyses of 25,000 scientific publications in various disciplines. Our regression analyses suggest that (1) interdisciplinary collaboration between domain scientists and computer scientists as well as the engagement of interdisciplinary individuals who have expertise in both domain and computer sciences are common in ML-related projects; (2) the engagement of interdisciplinary individuals seem more important in achieving high impact and novel discoveries, especially when a project employs computational and domain approaches interdependently; and (3) the contribution of ML and its implication to team structure depend on the depth of ML. Scientific knowledge shapes the foundation of the modern society, contributing to economic, social, and technological progress , 2. The Increasing examples have been reported in various domains, in which machines work alongside humans to push forward the progress of science. For example, in life sciences, protein-protein interactions are predicted to understand disease mechanisms ; in chemAs contemporary science is usually based on a team activity , the intdomain science" hereafter). Namely, our analysis includes six domains–agriculture, biology, chemistry, material sciences, medicine, and physics. To highlight the role of machines, we exploit a comparative approach, contrasting (1) ML-related projects  and (2) ML-unrelated projects (pure domain sciences). We collected approximately 2,500 ML-related and 22,000 ML-unrelated scholarly publications. With bibliometric and text analyses, we operationalized key variables of our interest. We investigated the quality (citation impact and novelty) of publication output produced by research teams with different team characteristics. Our results suggest (1) that interdisciplinary collaboration between domain scientists and computer scientists as well as the engagement of interdisciplinary individuals who have expertise in both domain and computer sciences are common in ML-related projects, (2) that the engagement of interdisciplinary individuals seem more important in achieving high impact and novel discoveries, especially when a project employs computational and domain approaches interdependently, and (3) that the contribution of ML and its implication to team structure depend on the depth of ML, in particular deep learning being associated with greater impact but with lower novelty.To this end, we draw on bibliometric analyses. Our primary interest is how computational science techniques are integrated into the fields of conventional domains of science : As another route to tackle interdisciplinary research challenges, individual scientists may acquire expertise in multiple domains. If an individual scientist has skills both in computer science and in domain science, the aforementioned organizational challenges can be resolved within him/herself. In fact, some areas of domain science recognized the promising power of computational techniques and actively incorporated computational science, such as bioinformatics in the biology domain . These dSuch interdisciplinary individuals can also play a boundary spanner role, who mediates members of different expertise , 49. TheThe engagement of an interdisciplinary scientist who has expertise in both computer science and domain science is positively associated with the quality of research output.Hypothesis 2 (H2): Among these organizational arrangements, the optimal form may depend on how computer science expertise is applied to domain science, or on the interdependency of the two areas of tasks . On the The proximity of collaboration between computer scientists and domain scientists is more positively associated with the quality of research output when the computer-related tasks and domain tasks are interdependent.Hypothesis 3A (H3A): The engagement of an interdisciplinary scientist who has expertise in both computer science and domain science is more positively associated with the quality of research output when the computer-related tasks and domain tasks are interdependent.Hypothesis 3B (H3B): To test our hypotheses, we draw on bibliometric data collected from the Web of Science (WoS). Our primary interest is in the integration of computational science (ML) and domain science. To highlight the contribution of ML, we draw on a comparative approach, contrasting (1) ML-related projects (combination of computation and domain sciences) and (2) ML-unrelated projects (purely domain science). The unit of analysis is a project team, which is operationalized by a group of authors of a publication.We employ the following sampling strategy. First, we chose six domains–agriculture, biology, chemistry, material sciences, medicine, and physics. The selection of these domains is based on WoS Subject Categories (SC). We chose 20 SCs in total within these domains see .Second, in these domains (SCs), we aimed to choose journals that are as mono-disciplinary as possible for two reasons. First, this is to lower the risk of sampling ML-related projects that are unrelated to domain science. We assume that mono-disciplinary journals set a clear scope of publication, with such a risk being mitigated. Second, to clarify the impact due to the integration of computer science and domain science, we minimized noise stemming from interdisciplinarity within a domain. To these ends, we chose up to five journals in each SC that are associated with a single SC (not associated with any other SC).Third, in these journals we selected two sets of papers, ML-related and ML-unrelated. We first searched for ML-related papers with "machine learning", "deep learning", and "artificial intelligence" as search keywords, which resulted in 2,500 papers.Next, we collected ML-unrelated papers that include none of the ML-related keywords. For clearer comparison, for each ML-related paper, we randomly selected up to 10 ML-unrelated papers published in the same journal and in the same year. We found 22,300 ML-unrelated papers. In total, we sampled 25,000 papers with 10% of ML-related papers and downloaded their bibliometric information.Impact).As the dependent variables of our analyses, we prepared two measures of scientific quality. First, we use the citation count as of 2021 to assess the impact of the findings reported in the paper. To mitigate the skewness, we took a natural logarithm of citation count (Novelty). We computed this variable only for part of our sample (67%) due to lack of access to citation network information.Second, we measure the novelty of a paper. This is because we are interested in to what extent ML contributes to creating new knowledge beyond human cognition. We drew on the recombinant novelty concept , 52 and Comp-Domain Collab).Since we are interested in the integration of computational expertise and conventional expertise, we investigated the forms of collaboration. To this end, we scrutinized the names of authors’ affiliated organizations to distinguish if an organization has a computational background or not. Then, we consider an organization to be computational if the name includes "computation", "information", or "system", and we consider an organization to be in a traditional domain if the name includes none of them. Using this distinction of organizations, we prepared a few variables. We first measured if a team involved both computational and domain organizations (instead of involving only domain organizations). A dummy variable is coded 1 if at least one affiliated organization is computational and 0 otherwise (Intra-Org Collab). Similarly, if a team involved computational and domain organizations in two different organizations, another dummy variable is coded 1 and otherwise 0 (Inter-Org Collab). Note that one team can involve both intra-organizational collaboration and inter-organizational collaboration.To further investigate the proximity in collaborating parties, we examined whether a team involved computational and domain organizations inside the same parent organization . In such teams, a dummy variable is coded 1 and otherwise 0 (Multi-Affiliation). The other measure is based on the previous experience of individual members. For feasibility, we focused on the corresponding author of each paper and tracked WoS Subject Categories (SCs) associated with their previous publications. We grouped SCs into computer-related and domain-related SCs, and coded a dummy variable 1 if the previous paper is associated with both a computer-related SC and a domain-related SC, and 0 otherwise (Multi-Expertise).Third, we measured whether an individual team member has both computational and domain expertise in two ways. One measure is based on organizational affiliation. If an individual member of a team (an author of a paper) is affiliated with both computational and domain organizations, we consider that the member has both computational and conventional expertise and plays a boundary spanner role at the individual level. A dummy variable is coded 1 if a team has at least one author affiliated with both types of organizations and 0 otherwise (Computer-Domain Integrated) should require a greater extent of integration between computer and domain expertise compared with the latter group (Computation-Focused).ML may be used in different ways in domain sciences. We sampled ML-related papers and categorized them into two groups by reading the method section of the papers. The first group of papers integrates both computational approaches and domain approaches , whereas the second group uses mainly computational approaches, typically based on secondary data. We assume that this is a critical distinction in that the former group (ML-related).Part of the following analyses compare ML-related papers and ML-unrelated papers. For this comparison, we prepared a dummy variable, coded 1 if a paper is ML-related  and 0 otherwise (DL-related).ML can mean various technologies. In fact, common technical keywords in our selected papers include, for example, "neural network", "classification", "regression", "support vector machine", and "random forest." Some of these have been used traditionally . To highlight the impact of machines, we attempted to differentiate the depth of models on which ML is carried out. Hence, we aim to contrast "deep" learning (DL) and "non-deep" learning (non-DL). This is because the distinction should affect the computer-science expertise required in a team and because deeper learning might provide new values for science beyond what traditional statistical techniques can do. Making this technical distinction based on text information is challenging because the detail of ML models is not always available. For feasibility, we consider that a project involved DL if a paper includes "deep learning" or "neural network" in the abstract or in the keywords (#Author), the number of organizations  included in the author address (#Org), and the number of countries included in the author address (#Country). We also control for university-industry collaboration as it is known to affect scientific performance [Univ-Industry Collab). We include publication year dummies as well as journal dummies. In so doing we control for systematic differences in the quality measures between publication years and between disciplines.In the regression analyses, we control for several variables. As the base characteristic of a team, we counted the number of authors of each paper . This may be because ML-related projects require less physical work and thus fewer members. The second bars, however, indicate that ML-related projects involve more organizations  probably because the ML-related projects tend to require a broader set of expertise . Finally, the bottom bars show no significant difference in the number of involved countries.We then describe the organizational features of ML-related projects as opposed to ML-unrelated projects. Then, we analyze the collaboration forms between computational and conventional organizations . As expeFinally, ML-related with controlling for the team size and other variables , suggesting that ML-related papers tend to receive more citations than ML-unrelated papers. On the other hand, Model 2 finds no significant coefficient of ML-related , suggesting that ML-related papers do not necessarily present novel discoveries. We further repeated the same set of analyses with a matching approach, in which ML-related papers are compared with ML-unrelated papers published in the same journal in the same year, finding a consistent result  and ones not involving such individuals (Comp-Domain Collab without Multi-Affiliation), finding that only the former group is associated with higher citation impact  but not the latter . This suggests that interdisciplinary individuals are important to achieve high citation impact. Similarly, Model 4 breaks down computer-domain collaboration into ones involving individuals having previous experience in computer and domain sciences (Multi-Expertise) and ones not involving such individuals (Comp-Domain Collab without Multi-Expertise). The result shows significantly positive coefficients for both variables , suggesting that high citation impact requires either computer-domain collaboration or interdisciplinary individuals. These results are supportive to H2.Model 3 further breaks down computer-domain collaboration into ones involving individuals affiliated with both computer and domain departments , which is consistent with H1, but they are both insignificant. Finally, Model 8 finds that interdisciplinary individuals having both computer and domain science expertise are weakly associated with higher novelty , consistent with H2.Multi-Affiliation and Multi-Expertise between ML-related and ML-unrelated projects (Multi-Affiliation) are significantly associated with higher citation impact , and Model 4 shows that individuals having both computer and domain expertise (Multi-Expertise) are significantly associated with higher novelty .These results seem to imply relatively greater importance of interdisciplinary individuals rather than interdisciplinary collaboration. Thus, we further test whether the role of interdisciplinary individuals is specific to ML-related projects. To this end, we compare the impact of projects . In all computation-focused projects and computer-domain integrated projects. We first test whether the proximity of collaboration shows a different impact between the two subsamples, finding mostly insignificant results. Thus, H3A is rejected. We then test whether interdisciplinary individuals make a greater impact on interdependent projects . However, Model 2 indicates that DL-related papers are less novel compared to DL-unrelated papers . Thus, it appears that DL does not necessarily allow scientists to gain novel insights beyond human cognition. DL rather seems to be applied to an agenda that humans have relatively good understanding of (and thus with lower novelty). Our interviewee suggested that DL tends to provide better model performance , facilitating further uses of DL models, which is consistent with the positive coefficient of DL-related on citation count (Model 1).Finally, we break down ML technologies. First, The progress of science increasingly relies on computational expertise and particularly on ML , and macThis study contributes to the literature by illustrating the role of machines in a scientist team. Previous literature on the use of machines has been either at a macro level  or at a A potential challenge in integrating computer and domain sciences is the lack of incentive. While our analysis shows that interdisciplinarity contributes to higher impacts and novelty, it is not obvious whether scientists with computational expertise and those with domain expertise are willing to work together. In fact, our interview suggested that scientists working on ML tended to appreciate publications in computer science rather than publications in other fields. It is also suggested that domain scientists do not always appreciate research approaches based upon ML because it is difficult to explain how prediction is made by a model that could be considered a black box. It is thus critical to understand what motivates ML scientists to collaborate with domain scientists and what obstacles exist in their collaboration.novelty of scientific discoveries with the engagement of interdisciplinary individuals. Our empirical work further suggests that interdisciplinary individuals are critical in delivering novel discoveries based on deeper ML (DL). ML can bring various values, such as the efficiency of data analysis or greater precision of model prediction. However, supplementing humans’ cognitive capacity and delivering novel discoveries is a fundamental benefit of machines under the burden of knowledge [It is particularly interesting to find that ML can contribute to the nowledge . Our resnowledge . EffortsOur results need to be interpreted with a few limitations. First, our bibliometric approach captures the structure of scientist teams only partially. Future research should look into how team members interact and what skills each member has. Second, future research should further investigate the technical aspects of ML. ML can be based on various models and can differ particularly in depth and complexity. We attempted to capture the depth by a simplified approach, but the result needs careful interpretation and a more sophisticated analysis is required. Third, we have to be cautious about potential changes in the role of machines over time. The vast majority of our sampled papers were published in the last four years (2017–2021) because ML is a rather recent phenomenon. The role of machines and how it affects the team design might change in the future with the advancement of computational techniques. Fourth, our results are based on cross-sectional analyses, and thus, the causal mechanism behind our findings cannot be completely clear.S1 File(ZIP)Click here for additional data file.S1 Appendix(PDF)Click here for additional data file.


Cellular lattices with architectural intricacy or metamaterials have gained a substantial amount of attention in the past decade due to the recent advances in additive manufacturing methods. The lattice topology controls its physical and mechanical properties; therefore, the main challenge is selecting the appropriate lattice topology for a desired function and application. In this work, we comprehensively study the topology–property relationship of three classes of polymer metamaterials based on triply periodic minimal surfaces (TPMS) of sheet/shell and ligament types, and other types of well-known strut-based lattices. The study uses a holistic approach of designing, additive manufacturing, microstructural characterization, and compressive uniaxial mechanical testing of these polymer lattices that are 3D-printed using the laser powder bed fusion technique known as selective laser sintering (SLS). In total, 55 lattices with different topologies and relative densities were 3D-printed and tested. Printing quality was assessed using scanning electron microscopy and micro-computed tomography. The extracted mechanical properties of elastic modulus, yield strength, plateau strength, and energy absorption are thoroughly compared between the different lattice architectures. The results show that all the investigated ligament-based TPMS polymer lattices exhibit bending-dominated elastic and plastic behavior, indicating that they are suitable candidates for energy absorbing applications. The sheet-based TPMS polymer lattices, similarly to the well-known Octet-Truss lattice, exhibited an elastic stretching-dominated mode of deformation and proved to have exceptional stiffness as compared to all other topologies, especially at low relative densities. However, the sheet-based TPMS polymer lattices exhibited a bending-dominated plastic behavior which is mainly driven by manufacturing defects. Overall, however, sheet-based TPMS polymer lattices exhibited the best mechanical properties, followed by strut-based lattices and finally by ligament-based TPMS lattices. Finally, it is depicted that at high relative densities, the mechanical properties of lattices of various architectures tend to converge, which implies that the topological effect is more significant at low relative densities. Generally, this study provides important insights about the selection of polymer mechanical metamaterials for various applications, and shows the superiority of TPMS-based polymer metamaterials as compared to several other classes of polymer mechanical metamaterials. Architected interconnected and cellular networks of solid struts, plates, or shells that form edges and faces of cells are known as metamaterials or latticed materials ,2,3,4. IThe most common types of cellular solids are stochastic foams that are considered a subset of cellular solids and were studied long before the attention shifted to architected or periodic cellular solids or lattices . However, the mechanical behavior of foams could be less reliable and hard to predict due to their random structures and possible extensive flaws and defects from manufacturability. To overcome such problems, lattice materials are designed to have an accurate estimate of their mechanical properties and consist of periodic unit cells ,16, wherManufacturing complex lattice architectures, especially with small feature sizes, is a fundamental challenge. However, recent developments in advanced macro/micro additive manufacturing (AM) techniques can overcome the limitations of conventional manufacturing techniques. AM is versatile and has been used to produce functional intricate parts using a broad spectrum of materials such as composites, metals ,56,57,58The polymer laser powder bed fusion technique known as selective laser sintering (SLS)  is consiIn the current literature, there are no systematic studies experimentally comparing polymeric cellular struts with ligament- and sheet-based TPMS lattices that are fabricated using SLS. In this regard, this paper compares the material–topology–property relationship of polymeric sheet-based TPMS, ligament-based TPMS, and cellular struts. This work extensively investigates the role of the topology in tuning the mechanical behavior by experimentally investigating and comparing the compressive mechanical properties of different sheet-based TPMS lattices, namely Schwarz Diamond, Schoen Gyroid, Schoen I-WP, and Fischer–Koch CY, with corresponding ligament-based lattices, and strut-based lattices with Gibson–Ashby (GA), Kelvin (K), Octet-Truss (OT), and Idealized Gyroid (IG) architectures at various relative densities.c is the level set parameter, which controls the solid volume fraction  in the TPMS lattice; Computer-aided design (CAD) files were created for different lattice architectures  technique, also known as polymer laser powder-bed fusion, using the Formiga P110 3D printer by EOS GMbH . The base material used for 3D printing was PA1102 black, which is a mass-colored black polyamide 11 powder , with siTM 250 FEG scanning electron microscope (SEM) to evaluate the 3D printing resolution. Moreover, the interior structure of the fabricated lattices was scanned using a micro-computed tomography (CT) scanner ), which has an ultra-high-performance nano-focus X-ray tube current with a wide sample and application range of ® visualization software (2019 version).After fabrication, architectural characterization was performed using the QuantaThe compressive mechanical properties of the 3D-printed lattices were obtained by performing uniaxial compression tests using the Universal Material Testing Machine Instron 5980 Series with a  ignored ,76. FiguThe designed RD is compared to the measured RD to assess the printability of the fabricated lattices. The measured RD is deduced by dividing the density of the lattice by the density of the base material, 99 g/cm3 . The denIt is also worth noting that the sheet-based lattices have higher surface area than strut-based lattices. In the context of the previous conclusion, more powder is most likely to fuse on the surface of the lattices. This could be attributed to the thermal gradient between the melted element and the loose powder nearby. We can say that the higher the surface area, the higher the deviation; this is also concluded by Al-Ketan et al. . AdditioExamples of SEM images of all the topologies are shown in The effect of varying the relative density for each lattice on its uniaxial compressive behavior is comprehensively investigated experimentally. The compressive stress–strain responses at different RDs are portrayed in Taking a closer look at the deformation mechanism of the 3D-printed lattices, it is noted that the deformation pattern is influenced mainly by the lattice topology. The deformation patterns of the different lattices are presented in The compressive mechanical properties of each lattice structure at each RD are compared with respect to the uniaxial modulus, yield strength, compressive plateau strength, and toughness, as illustrated in tectures . Thus, nIt can be noticed from n exponents of the uniaxial modulus for the D-STPMS and the Octet-Truss have the lowest values of 0.75 and 0.76, respectively, indicating much less dependency on RD. A comparable n exponent for the D-STPMS of 0.8 was attained by Bobbert et al. [n value of 0.5 for the D-STPMS and 1.23 for the Octet-Truss. The G-STPMS and IWP-STPMS have approximately the same n exponent of 1, where Abueidda et al. [n exponent varies between 1 and 2, i.e., 1.85. The ligament-based TPMS, Gibson–Ashby, and Idealized Gyroid lattices exhibited a bending deformation mode since the value of the n exponent is more than 2. This is in contrast to the conclusions of the Maraging steel SLM-printed topologies [n exponents of 1.68 and 1.62, respectively, implying a mixed deformation mode. Yan et al. [Following the sheet-based TPMS lattices are the common strut-based lattices, where the cellular struts showed enhanced mechanical properties in comparison to the ligament-based TPMS lattices. In terms of stiffness and yield strength, the sheet-based TPMS and the Octet-Truss lattices exhibited the best mechanical behavior at low relative densities when compared to other lattice topologies. The t et al. , whereasa et al.  concluden et al.  have stan et al. , rubber-n et al. , calciumn et al. , and phon et al. ,35,46. Tn value observed is higher than 1.5, implying that at yielding, the topologies adopted a bending-dominated deformation mode for all the architectures, apart from the G-STPMS and Octet-Truss lattices, which exhibited a stretching-dominated deformation mode behavior with an n value of approximately n exponent is equal to 1.25, while all the other architectures and even the sheet-based TPMS lattices exhibited a bending deformation mode, as all the n values are more than 1.5. This is in agreement with other studies on metallic TPMS lattices, where it was concluded that the compressive strength of the sheet-based TPMS lattices exhibits a bending-dominated mode of deformation for the Diamond, IWP, and Gyroid lattices [In terms of the yield strength, the lattices  showed tlattices  concludeIt is noteworthy that the toughness of bending-dominated lattices is usually larger than that of stretching-dominated lattices . In factIt is noteworthy that although the G-Idealized strut-based lattice is an idealization of the G-LSTPMS ligament-based lattice, overall, the G-Idealized lattice slightly outperforms the G-LSTPM lattice, especially at lower relative densities. Their elastic modulus and yield strength are almost the same, but the plateau strength and toughness of the G-Idealized lattice are slightly higher than that of G-LTPMS. In fact, one would expect G-LTPMS to perform better than G-Idealized due to its smooth surface with no geometric discontinuities. This counter-intuitive result can be mainly attributed to more manufacturing defects in G-LTPMS as compared to G-Idealized, as these defects affect the strength and toughness more than modulus and yield strength. Furthermore, while the results of the ligament-based TPMS lattices indicate that the G-LTPMS had superior mechanical properties among ligament-based lattices and is more promising at low relative densities, the D-LTPMS and CY-LTPMS had the lowest mechanical properties, which could be attributed to their smaller cross-sectional area at many junction points within the lattice, making them more prone to manufacturing defects.Generally, it can be noticed that at high relative densities, the compressive mechanical properties start to converge for different lattice topologies as the effect of the base material starts to dominate. Moreover, Kelvin lattice shows mechanical properties as high as the STPMS lattices at high relative densities. Although Kelvin lattice exhibits a bending-dominated deformation behavior, it can be observed that it has the highest compressive strength at high relative densities. This could be attributed to the hardening effect, which increases as the relative density increases, such that at high relative densities, a plateau in the stress–strain response is not observed and the strength continues to increase with inelastic deformation. On the other hand, the Octet-Truss lattice initially fails, and then the stress–strain response reaches a plateau. In fact, Kelvin lattice becomes much stronger at higher relative densities as the strut thickens and seems like it is converging similar to a closed unit cell, in that its resistance to load increases significantly and starts to behave like a sheet-based lattice. Based on the previous analyses of the mechanics of lattice materials, lattices with a stretching-dominated deformation behavior are more desirable in the design for additive manufacturing as they offer superior mechanical properties at the same relative density when compared to the bending-dominated topologies. Therefore, stretching-dominated lattices such as the Octet-Truss lattice have been a matter of extensive research ,87,88,89The stiffness and strength of the lattice architectures tested here are compared with other materials in We comprehensively studied the topology–property relationship of three classes of architected cellular materials or metamaterials based on TPMS lattices of ligament (LTPMS) and sheet/shell (STPMS) types and other types of well-known strut-based lattices additively manufactured using polymer laser selective laser sintering (SLS). In total, 55 lattices  were 3D-printed and tested under quasi-static compressive loading. The printing quality of these lattices was evaluated using SEM and micro-CT. Furthermore, detailed comparisons were made between the extracted compressive mechanical properties  of each lattice class of different topologies and relative densities. Overall, the results show that the stress responses of lattices 3D-printed using the SLS technique have smooth transition from elastic to plastic behavior, with a smooth hardening curve with increased deformation, except for the Octet-Truss lattice. Furthermore, STPMS lattices demonstrated a stretching-dominated elastic deformation behavior and a bending-dominated plastic behavior, and the latter deformation mode is mainly driven by manufacturing defects. Overall, however, STPMS lattices exhibited the best mechanical properties, with D-STPMS and G-STPMS having the best mechanical performance, followed by strut-based lattices and finally LTPMS lattices. Kelvin strut-based lattice exhibited high compressive strength and toughness that are comparable to STPMS lattices, especially at high relative densities. An interesting result is that the G-Idealized lattice, which is a strut-based idealization of the smooth G-LTPMS ligament-based lattice, exhibits slightly higher plateau strength and toughness than G-LTPMS at lower relative densities, which can be attributed to more manufacturing defects in the 3D printing of G-LTPMS.n scaling exponents for various properties. However, it is noticed that at high relative density, the mechanical properties of all the topologies tend to converge, implying that the effect of topology is more significant at low relative densities. Furthermore, all the investigated ligament-based TPMS lattices demonstrated a bending-dominated deformation mode, indicating that they are best for energy absorption applications. This study emphasizes the attractive mechanical behavior of sheet/shell-based lattices as compared to strut/ligament-based lattices and their utilization in various applications.Overall, the D-STPMS and Octet-Truss lattices exhibited the best compressive mechanical properties, as they are the least affected by the change in the relative density because of their small Gibson–Ashby


Effects of B-vitamin/folate therapy on CLT and Absmax were studied. Plasma samples were collected from 1,952 CAD patients randomized in a 2 x 2 factorial design to (i) folic acid, vitamins B12, B6; (ii) folic acid, vitamin B12; (iii) vitamin B6; (iv) placebo for 3.8 years in the Western Norway B-Vitamin Intervention Trial. Clot lysis time (CLT) and maximum absorbance (Absmax) were determined using a validated turbidimetric assay. Acute myocardial infarction (AMI) and mortality were assessed during a 7-year follow-up. Data were analyzed using bivariate and multiple regression. Survival free of events was studied using Kaplan Mayer plots. Hazard ratios (HR) and 95% confidence intervals (CI) were estimated using Cox proportional hazards models. Baseline urinary homocysteine (uHcy)-thiolactone and plasma cysteine (Cys) were significantly associated with CLT while plasma total Hcy was significantly associated with Absmax, independently of fibrinogen, triglycerides, vitamin E, glomerular filtration rate, body mass index, age, sex plasma creatinine, CRP, HDL-C, ApoA1, and previous diseases. B-vitamins/folate did not affect CLT and Absmax. Kaplan-Meier analysis showed associations of increased baseline CLT and Absmax with worse outcomes. In Cox regression analysis, baseline CLT and Absmax (>cutoff) predicted AMI  and mortality . After adjustments for other prognostic biomarkers these associations remained significant. Cys and uHcy-thiolactone, but not tHcy, were significant predictors of AMI in Cox regression models that included CLT. Conclusions uHcy-thiolactone and plasma Cys are novel determinants of CLT, an important predictor of adverse CAD outcomes. CLT and Absmax were not affected by B-vitamin/folate therapy, which could account for the lack of efficacy of such therapy in CAD.Fibrin clot structure/function contributes to cardiovascular disease. We examined sulfur-containing metabolites as determinants of fibrin clot lysis time (CLT) and maximum absorbance (AbsTrial registration: URL: http://clinicaltrials.gov. Identifier: NCT00354081. Thrombotic events initiated by an underlying vascular dysfunction are a major component of cardiovascular disease (CVD). In occlusive arterial disease the formation of a platelet-rich thrombus is supported by a fibrin mesh, whose formation depends on complex interactions between the components of the coagulation cascade. Accumulating evidence suggests that fibrin clot structure and function is associated with the development and progression of CVD. For example, dense structure of the fibrin clot, reflected in increased maximum absorbance and longer clot lysis time, have been observed in CVD patients . Fibrin i.e. hyperhomocysteinemia (HHcy), increases a risk for the development of CVD and stroke [S-Hcy-albumin, S-Hcy-IgG, and Hcy-S-S-Cys, with free reduced Hcy contributing only about 1% [N-Hcy-protein [Elevated plasma total homocysteine (tHcy), d stroke , 4. tHcyabout 1% , 4. Howe-protein , which h-protein  and stro-protein , are not-protein .N-Hcy-protein [Cbs-/- mice [12, and B6 vitamins did not affect levels of uHcy-thiolactone and its association with AMI [The only known source of Hcy, an important intermediate in one-carbon metabolism in humans, is the dietary protein methionine. Hcy is metabolized to Hcy-thiolactone in a reaction catalyzed by methionyl-tRNA synthetase. Hcy-thiolactone, a chemically reactive thioester, modifies protein lysine residues generating -protein . Hcy-thi-protein  than in -protein . Urinary-protein , and 10–max, a measure of clot structure) in a cohort of CAD patients participating in the Western Norway B-Vitamin Intervention Trial (WENBIT). We have studied associations of fibrin CTL and Absmax with Hcy-thiolactone, Hcy and cysteine (Cys), and effects of folic acid and B-vitamin therapy on these variables. We also examined how these associations are influenced by other variables related to CVD risk and examined prognostic values of CLT and Absmax.How sulfur-containing metabolites affect fibrin clot structure/function has not been examined in large randomized clinical trials. For this reason, we quantified fibrin clot lysis time  and maximum absorbance (Absi) folic acid (0.8 mg) + vitamin B12 , vitamin B6 ; (ii) folic acid + vitamin B12; (iii) vitamin B6; (iv) placebo for an average of 3.8 years. The study medication  was given as a single capsule, indistinguishable by color, weight, or the ability to dissolve in water.We analyzed existing citrated plasma samples from patients with suspected CAD who underwent coronary angiography for stable angina pectoris and participated in WENBIT . ParticiThe present study included 61.2-year-old patients  from baseline  and the end-of-study (n = 192) for whom uHcy-thiolactone values were available. Samples were assayed by investigators blinded to the clinical data to avoid bias. Written informed consent was obtained on the day of randomization. The study protocol was in accordance with the principles of the Declaration of Helsinki and was approved by the Regional Committee for Medical and Health Research Ethics, Bergen, Norway; by the Norwegian Medicines Agency, Bergen, Norway; and the WENBIT Steering Committee, Bergen, Norway.https://cvdnor.b.uib.no/) project, which provided information on discharge diagnoses from Norwegian hospitals during 1994–2009, linked to each patient’s unique 11-digit personal number [The endpoints were mortality and AMI, which included both fatal and nonfatal events, defined according to the International Classification on Diseases (ICD) 10th edition; I21-22. Information on endpoints was obtained from the Cardiovascular Disease in Norway  to each well of the 96-well plate using a multichannel pipette at 20 sec intervals. The time of addition of the activation mix was recorded to enable the plate reader times to be adjusted to the start of clot initiation. Absorbance was read at 340 nm every 30 s for 1 hour in a Tecan NanoQuant Infinite M200 Pro microplate reader. Complete lysis of fibrin clots occurred within 1 hour at the tPA concentration used. Plasma sample were assayed in duplicates and the values were averaged.The assay was modified from that described previously . BrieflyS1 Fig, were analyzed using a customized software kindly provided by Dr. Peter Grant [max, a measure of fibrin network density) and fibrin CLT  were calculated from the kinetics. Inter-assay variabilities for Absmax and fibrin CLT were 1.5% and 4.7%, respectively. The definitions of clotting and lysis variables are shown in S1 Fig. Correlations between the clotting and lysis variables in the WENBIT cohort, shown in S1 Table, are like the correlations previously reported in healthy individuals [Kinetics of fibrin clot formation and lysis, illustrated in er Grant . Maximumividuals .6 and B12, urinary Hcy-thiolactone and creatinine, and other variables, determined by standard assays, were obtained from analyses reported previously [Values for serum/plasma tHcy, Cys, creatinine, folate, vitamins Beviously . Hcy-thit-test was used for comparisons between two groups of variables with normal distribution. A Mann-Whitney rank sum test was used for comparisons between two groups of non-normally distributed variables. Associations between fibrin clot lysis time (CLT) or Absmax and other variables were studied by bivariate and multiple regression analyses. Event-free survival was analyzed by the Kaplan-Meier method and log-rank test was used to estimate differences in survival between patients with long vs. short fibrin CLT. Hazard ratios (HR) and 95% confidence intervals (CI) for clinical events associated with long vs. short fibrin CLT were calculated using the multivariable Cox proportional hazard regression analysis. The optimal cut-off values for CLT and Absmax were determined using a recursive partitioning statistical method that builds classification and regression trees for predicting continuous dependent variables (regression) and categorical predictor variables (classification) (https://statisticasoftware.wordpress.com/2012/06/15/classification-and-regression-trees/). Statistical software packages Statistica, version 13  and PSPP, version 1.0.1 (www.gnu.org) were used. Probability values were 2-sided and P value <0.05 was considered significant.Normality of distribution was tested with the Shapiro-Wilk’s statistic. Mean ± standard deviation (SD) or median was calculated for normally or non-normally distributed variables, respectively. An unpaired two-sided Table 1). Longer fibrin CLT in women was accompanied by reduced levels of uHcy-thiolactone, urinary creatinine (uCreatinine), plasma creatinine (pCreatinine), plasma tHcy (but not Cys), and older age compared to men. Fibrin clot Absmax varied from 0.0025 to 0.362 A340 and showed a tendency for lower values in women compared to men . Descriptive statistics of all variables analyzed in the present study are shown in S2 Table.For the CAD patients  mean age at baseline was 61.2 years and 71.5% were men. Baseline fibrin CLT varied from 105 to 1560 s and was significantly longer in women (n = 556) than in men (n = 1396), P = 0.0097 , GRF  and positively with plasma Cys  but was unaffected by tHcy levels . There was no correlation between fibrin Absmax and uHcy-thiolactone, GFR, plasma Cys or tHcy (not shown).Relationships between plasma fibrin CLT and uHcy-thiolactone, glomerular filtration rate (GFR), plasma Cys and tHcy are illustrated in These findings suggest that better clearance of plasma Hcy-thiolactone into urine  shortensS2B Fig). We also found that excretion of uHcy-thiolactone was better at high GFR and worse at low GFR , mimicking better excretion of uCreatinine at high GFR and worse at low GFR . Although we were not able to quantify plasma Hcy-thiolactone due to limited availability of samples, we found a negative correlation between plasma Cys and GFR , like the correlation between pCreatinine and GFR .These findings also suggest that impairments in kidney function and attenuation of GFR would reduce fibrinolysis. Indeed, fibrin CLT was shorter at high GFR and longer at low GFR . Methionine did not affect fibrin CLT or Absmax (not shown).Multiple regression analysis showed that fibrin CLT and Absmax were both associated with fibrinogen, albumin, and triglycerides (TG). However, while fibrin CLT and Absmax were positively associated with fibrinogen, CLT and Absmax showed opposite associations with albumin and TG: positive for CLT and negative for Absmax (Table 2).Fibrin CLT and Absmax showed disparate associations with other variables in multiple regression analysis. For example, fibrin CLT, but not Absmax, was associated with vitamin E, BMI, GFR, TG, and fibrinogen. On the other hand, Absmax, but not CLT, was associated with pCreatinine, CRP, HDL-C, and ApoA1 (Table 2).Fibrin CLT and AbsTable 2). Similarly, Absmax was associated with BMI, GFR, and Lpa in bivariate analysis, but not in multiple regression analysis. Conversely, associations of Absmax with tHcy, age, and sex found in multiple regression analysis, were not observed in bivariate analysis.Many associations found in bivariate analysis were not observed in multiple regression analysis. Specifically, although fibrin CLT was associated with uCreatinine, CRP, HDL-C, ApoA1, Lpa, ApoB, and age in bivariate analysis, these associations were not observed in multiple regression analysis (Table 2). Inclusion of previous disease status in the multiple regression models did not affects these associations. In contrast, the association of tHcy or Cys with fibrin CLT lost significance in models adjusted for pCreat or uCreat, respectively (Table 2). The association of vitamin E with fibrin CLT lost significance in models adjusted for HDL-C and LDL-C. Additional adjustments for CRP, ApoB, ApoA1, Lpa, tCys/tHcy had no effect on these associations.The associations of uHcy-thiolactone, vitamin E, and albumin with fibrin CLT persisted in multiple regression models adjusted for age, sex, pCreatinine, and uCreatinine, as did the associations of BMI, GFR, and triglycerides with CLT (max (>0.025) (S3 Table). Kaplan-Meier analysis showed a worse survival free of AMI events among CAD patients with a longer fibrin CLT (>397.5 s) compared to patients with a shorter fibrin CLT (≤397.5) (P = 0.011) . Survival free of mortality was also worse in CAD patients with a longer fibrin CLT (>532.5 s) compared to patients with a shorter fibrin CLT (≤532.5 s) (P = 0.002) . Kaplan-Meier analysis also showed worse survival without an AMI  or mortality  among CAD patients with a higher Absmax compared to those with lower Absmax. These differences started early in year 1 and progressively increased in years 2–7 during the follow-up.During the median follow-up time of 7 years, there were 8.0% (n = 160) acute myocardial infarctions (AMI) and mortality was 5.8% (n = 116), with a higher proportion of events in groups with a longer plasma fibrin CLT (>397.5 s) or higher AbsP = 0.013) and mortality  . The association between fibrin CLT and AMI remained significant after adjustment for vitamin E , glucose and BMI , LDL-C, HDL-C, TG, APOA1, APOB, and Lpa , uHcy-thiolactone, tHcy, and Cys , and uHcy-thiolactone, tHcy, Cys, uCreatinine, pCreatinine, and GFR . However, adjustments for smoking, diabetes, hypertension, extent of CAD at angiography, LVEF, heart failure, previous peripheral artery disease, AMI, stroke, and coronary artery bypass attenuated the association between CLT and AMI , as did adjustments for fibrinogen and CRP .Multivariable Cox proportional hazard regression analysis in a model including age and sex, showed that fibrin CLT  was significantly associated with the incidence of AMI . Cys was also a significant predictor of mortality in Cox regression models that included fibrin Absmax.Cys and uHcy-thiolactone, but not tHcy, were significant predictors of AMI in Cox regression models that included fibrin CLT and sulfur-containing metabolites  was significantly associated with the incidence of AMI  and mortality  . The association between fibrin Absmax and AMI remained significant after adjustment for vitamin E , glucose and BMI , LDL-C, HDL-C, TG, APOA1, APOB, and Lpa , uHcy-thiolactone, tHcy and Cys , and uHcy-thiolactone, tHcy, Cys, uCreatinine, pCreatinine, and GFR . Adjustments for smoking, diabetes, hypertension, extent of CAD at angiography, LVEF, heart failure, previous peripheral artery disease, AMI, stroke, and coronary artery bypass  and for fibrinogen and CRP  did not affect the association between Absmax and AMI.Cox regression analysis in a model including age and sex, showed that fibrin Abs12 and/or vitamin B6 resulted in significant increases in plasma levels of these vitamins and a significant decrease in plasma tHcy at the end of study (Table 4). However, we found that the supplementation with any combination of folic acid and B vitamins did not affect fibrin CLT or Absmax (Table 4). This finding indicates that the reduction in plasma tHcy does not affect fibrin clot structure/function and is consistent with the absence of correlation between fibrin CLT or Absmax and plasma tHcy at baseline . We also found that uHcy-thiolactone and plasma Cys, which were correlated with fibrin CLT at baseline , were not affected by the supplementation with any combination of folic acid and B vitamins (Table 4).As previously reported , supplemi) uHcy-thiolactone was significantly associated with CLT, while plasma tHcy was a significantly determinant of Absmax, independent of fibrinogen, triglycerides, vitamin E, GFR, independently of fibrinogen, triglycerides, vitamin E, creatinine, CRP, HDL-C, ApoA1, GFR, BMI, age, sex, and previous disease status; (ii) Kaplan-Meier analyses showed worse survival and worse survival free of AMI in CAD patients with longer baseline CLT and higher Absmax; (iii) in Cox regression analyses, longer baseline CLT and higher Absmax predicted future AMI events and mortality; (iv) in Cox regression models with CLT, Cys and uHcy-thiolactone predicted AMI, while Cys predicted AMI in Cox models with Absmax. (v) B-vitamin/folic acid therapy did not affect CLT and Absmax.We found that at baseline,  in a cohort of CAD patients, consistent with previous findings in studies with healthy probands [Accumulating evidence suggests that dysregulation of sulfur amino acid metabolism and impaired fibrin clot properties are associated with CVD. In the present work we examined Hcy-thiolactone, Hcy, and Cys as determinants of fibrin CLT and Absprobands , 14.Table 2). The association of CLT with vitamin E is consistent with previous findings showing that vitamin E is an inhibitor of plasmin-mediated fibrinolysis [Notably, in multiple regression analysis, we found that uHcy-thiolactone was a negative predictor  while Cys was a positive predictor  of CLT. Association of uHcy-thiolactone with CLT was independent of vitamin E, which we identified as another new predictor of CLT  (inolysis .Table 2) as well as ApoB, Lpa, HDL-C, ApoA1 (not shown), known to be associated with CLT in bivariate analyses [Table 2). However, the association between plasma Cys and CLT was attenuated by the adjustment for uCreatinine. We found that tHcy was significantly associated with CLT only in an unadjusted model but not in models adjusted for age, sex, pCreatinine, and uCreatinine (Table 2). Another study with large cohorts of thrombosis patients (n = 770) and healthy controls (n = 743) also reported no association of CLT with tHcy [The association of CLT with uHcy-thiolactone was also independent of BMI, GFR, CRP, fibrinogen, and TG, known determinants of CLT –20. Impoanalyses –20, but ith tHcy .Table 2). Further, although we found that CRP, HDL-C, ApoA1, Lpa, and ApoB, but not tHcy, were significantly associated with CLT in a univariate analysis, these associations were absent in multivariate regression analysis (Table 2).Other studies reported that CLT was associated with tHcy , CRP 2323, Lpa , HDL-C amax with plasma tHcy  that has not been reported before. The association between tHcy and Absmax was not affected by other variables associated with Absmax in our cohort: albumin, TG, HDL-C; ApoA1, fibrinogen, pCreatinine, CRP, age, sex, and previous diseases. Adjustments for uHcy-thiolactone, Cys, BMI, GFR, uCreatinine, Lpa, LDL-C, and vitamin E did not affect the association between tHcy and Absmax. This finding suggests that tHcy may affect the structure of the plasma fibrin clot without affecting clot’s function (see below).Notably, in multiple regression analyses, we found a significant negative association of Absmax suggest that each metabolite can affect clot properties via different metabolite-specific mechanisms. Some metabolites may affect clot structure while others might affect function. For example, Hcy-thiolactone has the ability to modify protein lysine residues in a process called N-homocystinylation, which leads to protein damage [N-Hcy-protein [N-Hcy-protein levels, including N-Hcy-fibrinogen and N-Hcy-albumin [N-Hcy-fibrinogen is prothrombotic because it forms fibrin clots that are more resistant to lysis by plasmin [N-Hcy-fibrinogen and N-Hcy-albumin as well as S-Hcy-albumin and S-Cys-albumin have been identified in human plasma [The disparate effects of uHcy-thiolactone and tHcy on CLT and Absn damage . The acc-protein . More ef-protein  will red-albumin . N-Hcy-f plasmin . Hcy and plasmin . Indeed,n plasma .e.g., N-Hcy-fibrinogen, N-Hcy-albumin, S-Hcy-albumin), as well as variations in sulfur-containing metabolites , will result in fibrin clots of different composition, structure, and susceptibility to lysis. These factors most likely account for the associations of the sulfur-containing metabolites as well as fibrinogen and albumin with measures of fibrin clot properties such as CLT and Absmax.As fibrin clot contains other protein components, in addition to fibrinogen, variations in the levels of these proteins, their modified forms  and confirmed  our previous finding that uHcy-thiolactone is a predictor of AMI [This interpretation is consistent with our present findings showing that CLT and Absr of AMI .max predicted AMI (Table 3). Previous studies also have shown a positive association between plasma fibrin clot properties and outcomes. For example, a case-control study with AMI patients (n = 800) and controls  showed that long CLT and high Absmax (>90th percentile in controls) were associated with an increased risk of AMI, 2.62- and 1.66-fold, respectively [In the present study, we found that longer CLT and higher Absectively .th percentile) was associated with a 2.52-fold increased risk of major adverse cardiovascular events  at a 12-month follow-up [max predicted AMI and mortality in a large PLATO study involving patients with acute coronary syndrome  [max predicted AMI (n = 160 events) and mortality (n = 116) during a 7-year follow-up (Table 3). In a relatively small study with CAD patients , CLT or Absmax did not predict vascular events after a 3-year follow-up [Another study with 300 patients hospitalized with acute coronary syndrome found that long CLT (>77ollow-up . CLT andleeding)  and patiollow-up , possiblN-Hcy-protein autoantibodies [max), an important pro-atherogenic factor.Previous studies have suggested that the failure of B-vitamin to attenuate Hcy-thiolactone levels , anti-N-tibodies , and inftibodies , 35 coultibodies . Our premax reflects the clot structure while CLT is a functional property that reflects clot’s susceptibility to lysis. High Absmax indicates a more compact structure of dense thin fibers characterized by low permeability and low susceptibility to lysis, while low Absmax indicates a less compact structure of loose thicker fibers characterized by high permeability and high susceptibility to lysis [max should also similarly affect CLT and vice versa. However, we found that factors affecting Absmax did not affect CLT and factors affecting CLT did not affect Absmax (Table 2). Specifically, in multiple regression analysis we identified variables that were associated with (i) only plasma fibrin CLT  and (ii) only with plasma fibrin clot Absmax . Only three variables were associated with both plasma fibrin CLT and Absmax  (Table 2).It is generally accepted in the field that Absto lysis . These smax suggests that variables that affect only CLT may act on a functional component of the fibrinolysis cascade without affecting Absmax, i.e., clot structure. A case in point is vitamin E, which affected CLT, but not Absmax, and is known to inhibit plasmin-mediated fibrinolysis [max apparently involve changes in the clot structure that have no functional consequence on CLT. Only a few variables were correlated with both structure and function of the fibrin clot (Table 2).The lack of congruence between associations of some variables with CLT and Absinolysis . On the The present study is the first to evaluate sulfur-containing compounds as determinants of fibrin clot properties in relation to future AMI events and mortality in CAD patients. Major strengths of the present study are the size , the prospective design, an extended 7-year-long follow-up, and an extensive information regarding baseline clinical/biochemical characteristics. Further, the high E-values for HR and lower CI suggest that our findings are robust to the presence of unmeasured confounders. Thus, these findings are likely to be reproducible in other populations. As our study was limited to a white elderly population with CAD, our findings remain to be confirmed in populations in other age groups and ethnic backgrounds.max but not of CLT. CLT and Absmax predict future AMI events and mortality in CAD patients but were not affected by B-vitamin/folate supplementation. Our findings also suggest that targeting sulfur-containing metabolites other than tHcy might be a useful therapeutic strategy to mitigate prothrombotic phenotypes that increase a risk of AMI and mortality.uHcy-thiolactone and plasma Cys are novel determinants of CLT. Plasma Hcy is a determinant of AbsS1 Figmax, are highlighted in bold.Variables examined in a greater detail in the present study, CLT and Abs(PPTX)Click here for additional data file.S2 FigP values are shown. uHcy-thiolactone was present in each sample and varied from 1.3 to 1,724.0 nM.Relationships between CLT and plasma Cys (A.), tHcy (B.), uHcy-thiolactone (C.) and GFR (D.). Spearman (PPTX)Click here for additional data file.S3 FigRelationships between GFR and uHcy-thiolactone (A.), uCreatinine (B.), plasma Cys (C.) and pCreatinine (D.).(PPTX)Click here for additional data file.S1 Tableet al. [P<0.000, except when indicated otherwise. The clotting and lysis variables are illustrated in S1 Fig. *Terms Absmax and CLT referring to terms MaxAbs and Lysis50MA, respectively, of Carter et al. [Values are from analyses of data for n = 1,983 patients. Nomenclature is after Carter et al. . P<0.000r et al. , have be(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table*Groups of patients without and with AMI or mortality events are indicated by 0 and 1, respectively.(DOCX)Click here for additional data file.