Ordinal traits are important complex traits in crops, while genome-wide association study (GWAS) is a widely-used method in their gene mining. Presently, GWAS of continuous quantitative traits (C-GWAS) and single-locus association analysis method of ordinal traits are the main methods used for ordinal traits. However, the detection power of these two methods is low.To address this issue, we proposed a new method, named MTOTC, in which hierarchical data of ordinal traits are transformed into continuous phenotypic data (CPData).Then, FASTmrMLM, one C-GWAS method, was used to conduct GWAS for CPData. The results from the simulation studies showed that, MTOTC+FASTmrMLM for ordinal traits was better than the classical methods when there were four and fewer hierarchical levels. In addition, when MTOTC was combined with FASTmrEMMA, mrMLM, ISIS EM-BLASSO, pLARmEB, and pKWmEB, relatively high power and low false positive rate in QTN detection were observed as well. Subsequently, MTOTC was applied to analyze the hierarchical data of soybean salt-alkali tolerance. It was revealed that more significant QTNs were detected when MTOTC was combined with any of the above six C-GWAs.Accordingly, the new method increases the choices of the GWAS methods for ordinal traits and helps to mine the genes for ordinal traits in resource populations. This includes count data for quantitative traits and hierarchical data for resistance traits, such as the number of main stem nodes , the numOrdinal traits are discrete traits that are controlled by multiple genes. However, their phenotypic data is hierarchical and non-continuous and contains relatively limited information; accordingly, GWAS for ordinal traits is more complex than that for continuous quantitative traits. The threshold model represents a reasonable method for the genetic analysis of ordinal traits, and most association mapping methods are developed under this framework . GeneralAccordingly, in this study, we proposed a method for transforming ordinal phenotypes into continuous phenotypes (MTOTC). First, the hierarchical phenotypic data for ordinal traits (HData) was transformed into continuous phenotypic data (CPData). Subsequently, FASTmrMLM , one GWA2Here we proposed a method, named MTOTC, to transform the discrete hierarchical data (HData) of ordinal traits into continuous phenotypic data. Then, GWAS for continuous quantitative traits (C-GWAS) are used to analyze the transformed continuous phenotypic data. The new method was described as below.2.1Arabidopsis thaliana lines harboring 10,000 SNPs with a minimum allele frequency >0.1 . This is MTOTC. Then, the predicted CPData is analyzed by C-GWAS methods, such as FASTmrMLM Figure 12.2.1P-value ≤0.05). To further improve the quality of the significant correlated SNPs in the initial screening for reducing interference and improving detection accuracy, logistic regression was used as a secondary SNP screening method. Logistic regression was performed using function “glm”  and “polr”  with a P-value ≤0.05. The aim of this step was to further eliminate SNPs that were not associated with the traits for simplifying the iterations in the following multi-locus genetic model.The Chi-square test in R 4.0.5 (function “chisq.test”) was used to scan the SNPs in the whole genome using a single marker method , set The parameters were estimated using empirical Bayes, as follows, and the Newton–Raphson method.Among them,Then, the empirical Bayesian estimates of these SNPs effects were obtained in the multi-locus model (1) based on the selected significant SNP markers and ordinal phenotype, and estimates of these effect were used to predict the phenotype, obtaining the continuous phenotypic data (CPData) of ordinal trait.2.3When continuous phenotypic data was obtained by the above MTOTC method, a C-GWAS method could be used to detect significant loci. In this work, FASTmrMLM, one C-GWAS method, was used. So loci significantly associated with ordinal traits were detected by FASTmrMLM using the obtained continuous phenotypic data and the potential associated markers identified in the above-described initial screening. The GWAS method is henceforth referred to as MTOTC+FASTmrMLM. Moreover, the effects of five other C-GWAS  methods are also discussed based on the MTOTC method for ordinal trait, in order to verify the feasibility of MTOTC.2.4We conducted six simulation studies to evaluate the feasibility of the new method. For each study, the loci 278, 2143, 2054, 3698, 1716, 6178, and 8501, located on chromosomes 1, 2, 2, 2, 1, 4, and 5, respectively, were selected as the causal loci related to the simulated trait. There were three types of phenotypic data in the simulation experiment—original data (OData), which were continuous and generated by Monte Carlo simulation; HData, which were generated from the above OData according to specific distribution proportions ; and CPData, which were generated from the above HData by MTOTC. Then, FASTmrMLM, one multi-locus C-GWAS algorithm, was used to conduct GWAS for CPData.33.13.1.1P-value >0.98 and an unusually large absolute value of effect estimate in logistic regression) were not truly related to the phenotype and interfered greatly with subsequent association analysis. Therefore, to further improve the quality of the screened significantly related SNPs and detection accuracy, logistic regression was used as a secondary screening method for SNPs in MTOTC.To determine the most suitable threshold value for the Chi-square test and logistic regression in the initial screening, four probability thresholds  were set for the Chi-square test in the first simulation study, while three probability thresholds  were set for logistic regression. The Chi-square test can eliminate a large number of SNPs that are not significantly related to a given phenotype. However, the simulation study showed that some SNPs screened in the above Chi-square test   was the lowest when the P-value was 0.0001, followed by 0.01, 0.05, and 0.10  when the P-value was 0.0001, followed by 0.01 and 0.05  Table 1.3.1.3The third simulation study investigated the effect of the number of hierarchical levels on MTOTC. Based on symmetrical distribution, the number of hierarchical levels was set to 2, 3, 4, and 5, respectively, and the number of replicates was 10,000. Meanwhile, we compared the results of OData, HData and CPData using FASTmrMLM.CV) of the OData was between 29.5% and 55.5%. Among the 10,000 replicates, the number of replicates beyond the CV range of the OData  also increased with increasing hierarchical level. Thus, the CV range of CPData2 was the closest to that of the OData. Third, among the 10,000 replicates, the skewness range between the CPData and the OData was the closest at 2 hierarchical levels. Among them, the skewness range of the OData was between −1.00 and 0.46 and the range of CPData2 was between −1.28 and 0.35. As the number of hierarchical levels increased, the skewness of the CPData gradually deviated from that of the OData; the kurtosis showed the same tendency as the skewness.Compared with CPData from the other hierarchical levels, the distribution of CPData2  was closer to the original data (OData). First, the frequency distribution of the CPData was closer to that of the OData when the hierarchical level was low, especially when it was equal to 2 Figure 4MTOTC performed well for the estimates of QTN position under different numbers of hierarchical levels. The position estimates via MTOTC+FASTmrMLM  were unbiased at loci 278, 2143, and 3698 , CPData3 (1:3:1), and CPData5 (1:2:4:2:1), 10 replicates were set at equal intervals from 1,000 to 10,000. As a result, the results across various numbers of replicates at each locus and for each hierarchical levels  were insignificant Figure 53.1.5In the fifth simulation study, we investigated the effect of distribution proportion skewness on the new method under three hierarchical levels. Here the distribution proportion skewness were set as symmetrical distribution , uniform distribution (1:1:1), and skewed distribution (4:2:1). The indicators were the relative power, false-positive rate, the estimates of QTN effects and positions. The skewed distribution had the lowest relative power at loci 278, 2143, and 3698, followed by the uniform distribution, and the symmetrical distribution  compared with the other five GWAS methods, which may be related to its relatively higher false-positive rate. Additionally, the numbers of significant QTNs detected by pKWmEB, mrMLM, and FASTmrMLM in CPData  were second only to the number of QTNs detected with pLARmEB.Four QTNs  for salt tolerance were simultaneously detected in the index data and at least one CPData; however, none of them was detected in HData5. For instance, Chr13-64738 was simultaneously detected in CPData2 by five methods and in the salt tolerance index data by two methods. Chr13-65248 was detected in CPData5 by four methods and in both CPData5 and the index data by FASTmrMLM. Three QTNs  for alkali tolerance were simultaneously detected in the index data and in at least one CPData, two of them were also detected in HData5.The results of six GWAS methods for the CPData of salt-alkali tolerance showed that only a few significant QTNs were coincident between 2009 and 2010, which can be explained by the differences in environmental influences between the two years. For salt tolerance, no QTNs were found to overlap between 2009 and 2010 in the six methods. For alkali tolerance, only Chr1-5051 and Chr16-82333 were detected in both years. There was indeed an environmental (year) effect according to variance analysis of the phenotypic results for the two years .3.2.2Error! Hyperlink reference not valid.http://www.Soybase.org/) was also used to screen candidate genes. A total of 34 potentially candidate genes for salt tolerance and 25 potentially candidate genes for alkali tolerance were mined.Potential candidate genes were mined from 100 kb upstream to 100 kb downstream  of signiGlyma05g25331, Glyma05g25420, Glyma05g25450, and Glyma05g25460 were all detected by five GWAS methods in CPData5 in 2009. Only one gene, Glyma13g25266, was detected in both the index data and CPData2 detected by five GWAS methods in CPData2 and two methods in the index data in 2010. In addition, five candidate genes were detected only in CPData2 by five methods, and nine candidate genes were detected only in CPData5 by three or more methods. No overlapping genes were found between HData5 and the index data or the CPData , between haplotypes TCAT and AATT (P=0.0091) in 2010, and between haplotypes TCAT and TCTT (P=0.0471) in 2010. However, for haplotype blocks of other salt tolerance QTNs, it was showed that the significant phenotypic differences existed between haplotypes only in a single year, and the haplotype pairs with significant differences included haplotype AGTGC and TACCC (P=0.0348), AGTGC and TGTCA (P=0.0345) for Chr5-24153; haplotype GCG and ATA (P=0.0408) for Chr10-52140; haplotypes GTAGA and GTAGT (P=0.0397), GTAGT and AAGTT (P=0.0540) for Chr11-54042.In haplotype block with the significant QTNs Chr13-64738 for salt tolerance, t-test showed significant phenotypic differences between haplotypes ACAT and AATT , between haplotypes CTGACG and CCGGAG (P=0.0005 in 2009), between haplotypes CTGACG and CCGAAG (P=0.0231 in 2009), between haplotypes TCGAAG and CCGAAG , and between haplotypes CCAAAG and CCGGAG (P=0.0296 in 2010). For Chr3-14262, the haplotype pairs with significant differences were detected as follows: TTT and TCT , TTT and GCT (P=0.0102 in 2010), GCT and TCT (P=0.0171). The other haplotype blocks of alkali tolerance showed significant phenotypic differences between haplotypes only in a single year and they include: GTGT and TTAT (P<0.0001), TTGT and TTAC (P=0.0038), TTAT and TAGT (P=0.0132) for Chr13-67342; CAG and TGT (P=0.0183) for Chr1-5051; ATCG and GATC (P=0.0009) for Chr7-34669; TAGGCG and AATGCA (P=0.0157), and TAGGCG and TATGCG (P=0.0128) for Chr20-105040.There were two significant QTNs Chr16-82333 and Chr3-14262 for alkali tolerance with significant phenotypic differences across haplotypes in both years. The Chr16-82333 recorded significant differences between haplotypes CTGACG and CCGGAG  and six alkali stress-related candidate genes  were found in the haplotype block.Genes with significant phenotypic differences across haplotypes were considered as the candidate genes 3, inclu4In this study, we established a method for transforming ordinal phenotypes into continuous phenotypes (MTOTC) based on hierarchical data for ordinal trait phenotypes and molecular marker data in resource populations. Therefore, the process of association analysis for ordinal traits is as follows: first, MTOTC is used to transform HData into continuous phenotypic data (CPData), and then a C-GWAS method (i.e. GWAS method for continuous quantitative traits) is selected to analyze the CPData to identify the QTNs that are significantly associated with ordinal traits.In this study, simulation experiments and soybean saline-alkali tolerance analysis indicated that the new method, MTOTC, is suitable for ordinal traits when they are less than five hierarchical levels. Moreover, the combination of MTOTC with any one of the proposed C-GWAs methods exhibited high power, low false-positive rates, and low bias in estimating the positions and effects of the QTN. The purpose of MTOTC is to provide a different approach for undertaking GWAS for ordinal traits. The feasibility of the MTOTC method was verified in real data analysis of soybean salt-alkaline tolerance using 286 soybean accessions. Compared with HData5 , a greater number of significant QTNs was detected concurrently by at least two GWAS methods or in two years, and more candidate genes for salt and alkali stress were screened in the CPData for salt and alkali tolerance traits. A greater number of QTNs was detected simultaneously by multiple GWAS methods in the CPData than in the index data and HData for salt-alkaline tolerance. For the three types of data, the number of QTNs detected simultaneously was respectively 4, 1, and 1 in salt tolerance and respectively 5, 2, and 3 in alkali resistance.When the phenotype distribution of the CPData generated by the new method were closer to those from the index data of salt-alkali tolerance, the GWAS results were better, and a greater number of candidate genes could be mined. This may be beneficial for selecting the appropriate distribution proportion to obtain hierarchical data of ordinal trait, screening stable QTNs, and promoting the development of molecular breeding. We also applied symmetric distribution (1:2:4:2:1) to generate HData5 for the salt tolerance index data and used MTOTC to generate the corresponding CPData5. The phenotype distribution of CPData5 with symmetric 1:2:4:2:1 exhibited a large deviation from that of the index data, and the phenotype distribution of CPData5 with uniform 1:1:1:1:1 was closer to that of the index data. Under the six methods, there were no overlapping QTNs in CPData5 and the index data for salt tolerance, which was far inferior to the above uniform distribution observed with the distribution proportion 1:1:1:1:1, under which three coincident QTNs were detected in CPData5 and the index data. This result corresponded precisely to the results presented in simulation study 5.P ≤ 0.05 threshold, a large number of SNPs that were significantly unrelated to the trait could be eliminated. Meanwhile, the simulation experiment showed that the retention rates of related loci remained high. MTOTC serves to simplify the model and save a substantial amount of computing time for subsequent association studies.MTOTC performed well in the initial SNP screening. After preliminary screening under a MTOTC helps to improve association analyses of ordinal traits. Regarding coefficient of variation, skewness, kurtosis, and frequency distribution, compared with the HData, the results obtained for the CPData were closer to those of the OData. Meanwhile, the results using six GWAS methods showed that the statistical power, the false-positive rate, and the position estimates in CPData were better than those in HData. Moreover, MTOTC performed better when the frequency distribution of the CPData was close to that of the OData.The fewer hierarchical levels, the more suitable MTOTC is. Regarding the relative power in CPData under different hierarchical levels, a trend of increasing relative power with increasing number of hierarchical levels was found for all six methods when there were four or less hierarchical levels. When there were five hierarchical levels, the power of MTOTC+FASTmrMLM was close to that of FASTmrMLM in HData, but slightly lower than the power from logistic regression; only three GWAS methods had higher relative power in CPData than in HData. In addition, MTOTC had a tendency to increase variation, especially with increasing numbers of hierarchical levels. This indicates that MTOTC is more suitable for ordinal traits with fewer hierarchical levels, especially those with two or three levels. Among the six GWAS methods, FASTmrEMMA, FASTmrMLM, and mrMLM are significantly better when combined with MTOTC. This is partly attributed to that the distribution and parameter estimation principles set in MTOTC were relatively consistent with those in these three GWAS models.This study will contribute to further research in association analysis of ordinal traits. This is especially in improving the retention rate of small-effect loci in preliminary screening, reducing the impact on variability when transforming ordinal phenotypes into continuous phenotypes, and developing novel methods for association analyses of ordinal traits.The original contributions presented in the study are included in the article/MY, JF, and YW designed the methodologies. MY, JF, JZ, and JCZ drafted the manuscript, conducted simulation studies, and analyzed the data. TZ and JF revised the paper. All authors contributed to the article and approved the submitted version.


The influence on the device of parameters, including perovskite thickness, total defect density, series and shunt resistances, and operating temperature, are discussed and analyzed. With optimized parameters, the efficiency increased from 13.47% to 18.33%. Then, a new 3NH3PbI3/PCBM/Ag device was proposed which includes a silicon-rich oxide (Inverted perovskite solar cells (PSCs) have gained much attention due to their low hysteresis effect, easy fabrication, and good stability. In this research, an inverted perovskite solar cell ITO/PEDOT:PSS/CH Perovskite solar cells (PSCs) have become a focus of research since they appeared in 2009 and are currently the most promising devices of the future. In just a few years, they have made significant progress, and it has been demonstrated that the power conversion efficiency . Pb is not environmentally friendly and has deleterious effects on humans and the environment. Consequently, numerous theoretical analyses have shown that inorganic perovskites based on carriers ,7.In addition, scientists are exploring the field of perovskite solar cells with different configurations. Some alternatives involve the replacement of or variations in electron and hole transport layers (HTLs and ETLs), the substitution of cations in the perovskite layer, and the use of intermediate or outer layers in the cells to achieve higher efficiency and improve the spectral response. Improving the utilization of the incident solar spectrum may be an approach to increasing the performance of perovskite solar cells because they currently have problems with their stability due to humidity and oxygen in the environment . Existinadiation . In addie values ,12.In this work, an inverted perovskite solar cell, ITO/PEDOT:PSS/The structure of the simulated inverted planar perovskite solar cell has the configuration ITO/PEDOT:PSS/ contact . MeanwhiThe simulation was performed in the SCAPS-1D software version 3.3.10 under AM1.5G illumination and at an ambient room temperature of 300 K. The SCAPS-1D software, developed by Professor Marc Burgelman of the Department of Electronics and Information Systems (ELIS) at the University of Ghent in Belgium, has been used to model and simulate perovskite solar cells . The sofThe physical, optical, and electrical parameters were obtained from a review of the scientific literature and are summarized in The simulated absorption coefficient of the perovskite active layer was obtained from SCAPS-1D software. We extracted the data and plotted them in Optimal selection of perovskite layer thickness is necessary to obtain a solar cell with efficient output values . Accordiion rate . The dec7) below .(7)Voc=The total defect density formance ,28. Recot values ,26. The t values .(8)RSRHConsequently, the minority carrier lifetime ectively .(9)τn,pow value .013 cm−3 ,28.On the other hand, the density of interface defects also plays an essential role in the efficiency of the solar cell due to defects functioning as recombination sites degrading the J-V and QE (%) output results . Interfadecrease . For exa013 cm−2 ,43.Series and shunt resistance (ar cells . To undewas used .(11)J=JIn an open-circuit state  and subsequently emits low-energy photons (visible light), where a solar device has the most advantages and sensitivity. The main characteristics of a down-conversion energy material are high PL photoluminescence in the visible spectrum, broadband absorption in the region where the solar cell response spectrum is low, a high absorption coefficient in the low wavelength region, high transmittance, broadband emission where the device response is high, chemical, and environmental stability, low roughness and finally ease of processing ,10. In tnnealing ,49. The Photoluminescence spectra were obtained using a Horiba Jobin Yvon NanoLog FR3 device and are shown in acancies ,51.Subsequently, transmittance spectra were obtained using a Varian Cary 5000 device and are shown in The transmittance spectra of the 4) below .(14)α  below .(15) or IPCE (incident-photon-to-current efficiency) for both devices b. EQE isAfterward, we obtained the value of the integrated photocurrent density ( devices a,b. The In addition, the SCAPS-1D software allows us to obtain the energy band diagram of the solar cell. In this work, we numerically simulated and optimized an inverted planar perovskite solar cell with ITO/PEDOT:PSS/In conclusion, the silicon-rich oxide


Using laser speckle flowgraphy (LSFG), we investigated the time course of changes in choroidal circulation hemodynamics after a cold pressor test in healthy eyes. This prospective study included the right eye of 19 young healthy participants. The macular mean blur rate (MBR) was measured with LSFG. The MBR, intraocular pressure (IOP), systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate (HR), mean blood pressure (MBP), and ocular perfusion pressure (OPP) were evaluated at baseline; immediately after the test and 10, 20, and 30 min later. Immediately after the test (0 min), SBP, DBP, MBP, and OPP were significantly elevated compared with those at baseline. The macular MBR significantly increased by +10.3 ± 7.1% immediately after the test. However, there was no change after 10, 20, and 30 min in the above parameter. A significant positive correlation of the macular MBR with the SBP, MBP, and OPP was observed. In young healthy individuals, increased sympathetic activity induced by a cold pressor test increases choroidal hemodynamics in the macula along with an increase in systemic circulatory dynamics, which normalizes after 10 min. Therefore, LSFG may provide a novel approach for assessing sympathetic activity and intrinsic vascular responsiveness in the eye. Stress caused by cold exposure activates the sympathetic nervous system, resulting in increased blood pressure (BP) and heart rate (HR); these are known as pressor responses, and cold pressor tests are performed to assess sympathetic responses in systemic circulatory dynamics ,2,3,4,5.The choroidal blood supply accounts for 85% of the total ocular blood flow . AutoregLaser speckle flowgraphy (LSFG) can be used to noninvasively measure choroidal blood flow and has the advantage of providing reproducible data throughout retinochoroidal diseases ,16,17,18Additionally, previous reports of a significant increase in choroidal blood flow velocity with increasing IOP, BP, and OPP after isometric exercise in healthy individuals indicate that choroidal circulatory dynamics are dependent on systemic circulatory dynamics ,11,20.Therefore, it is hypothesized that the choroidal circulations will also increase in conjunction with the increase in systemic circulations by the cold pressor test. This may lead to the prediction and early detection of retinochoroidal diseases and the classification of their severity. However, changes in choroidal blood flow with LSFG after an increase in sympathetic activity induced by a cold pressor test have not yet been assessed. In this study, we investigated the time course of changes in the MBR after a cold pressor test using LSFG.This study was approved by the ethics committee of Fukuoka International University of Health and Welfare  and adhered to the tenets of the Declaration of Helsinki. Written informed consent was obtained from all participants.This prospective study included the right eyes of 19 young healthy volunteers  with no ophthalmic or cardiovascular diseases . All parEach participant underwent examinations, including best-corrected visual acuity, fundus photography, IOP, and LSFG. SBP, DBP, and HR were also measured.The examinations were carried out in a quiet examination room at room temperature (24 ± 1 °C), and humidity was maintained at 47 ± 3%. During the cold pressor test, the right hand was immersed up to the wrist in ice water at approximately 1 °C for 30 s ,13. LSFGLSFG targets moving red blood cells in the deep choroidal vessels using an 830 nm diode laser to illuminate the ocular fundus. The laser speckle method is advantageous because it is quantitative and replicable, and its application to measure ocular blood velocity has been previously reported to yield reproducible results ,21,22. LThe measurement screen of LSFG (a). The MBR within the region of interest immediately after the cold pressor test (c) increased by 11.1% compared with that at baseline (b). However, after 10, 20, and 30 min of testing, the results were similar to those at baseline at −1.5% (d), −1.5% (e), and +0.8% (f), respectively.For all participants, BP and IOP were measured at baseline, immediately after the cold pressor test (0 min), and 10, 20, and 30 min later. The IOP measurement was performed with a noncontact tonometer . Hemodynamics in the eye depend on BP and IOP and are measured using an indicator called OPP, which reflects systemic circulatory dynamics. The OPP was calculated from the IOP and BP values . MBP wasOPP was calculated as:p values < 0.05 indicated statistical significance.All results are expressed as the mean ± standard deviation. The Friedman test and Scheffe’s paired comparison test were used to examine sequential changes in IOP, SBP, DBP, MBP, HR, OPP, and the MBR. Spearman’s rank correlation test was used to determine the relationships between the rate of change in MBR and other studied factors, including IOP, SBP, DBP, MBP, HR, and OPP. For all tests, The mean age was 22.3 ± 3.3 years . All participants had a best-corrected visual acuity score ≥ 20/20.Changes in the values of IOP and other systemic factors are summarized in p < 0.001, p = 0.025, p = 0.005, p = 0.004, p < 0.001, p < 0.001); they showed no changes after 10, 20, and 30 min.The mean IOP and HR did not change throughout the study  a,e. The The changes in the OPP are presented in MBR changes are presented in The correlation between the rate of change in MBR and changes in other studied factors from baseline to a point immediately after the cold pressor test (0 min) was examined. MBR showed a statistically significant positive correlation with SBP, MBP, and OPP . HoweverIn this study, SBP, DBP, MBP, OPP, and MBR increased significantly after the cold pressor test but did not vary further after 10 or more minutes. In addition, there was a statistically significant positive correlation of the macular MBR with SBP, MBP, and OPP. These results indicate that an increase in sympathetic activity induced by cold stimulation also increases the choroidal circulation hemodynamics, depending on systemic circulatory hemodynamics, and that these values normalize after 10 min.Previous reports have shown that both SBP and DBP increase owing to vasoconstriction caused by sympathetic hyperactivity in the cold pressor test ,2,3,4,5.It has been reported that choroidal blood flow velocity increases during the acute phase in sympathetic hyperactivity etiologies such as CSC. There are two possible causes of increased choroidal blood flow in eyes with acute CSC. Firstly, the activation of sympathetic α-adrenergic receptors causes vasoconstriction of the choroidal arteries and secondary overflow into the choroidal vasculature. Secondly, the cardiac output is increased owing to the activation of sympathetic β-adrenergic receptors, which increases blood flow in the choroid ,16. FurtAdditionally, previous studies showing a significant increase in choroidal blood flow velocity with increasing IOP, BP, and OPP after isometric exercise in healthy individuals indicate that choroidal circulatory dynamics are dependent on systemic circulatory dynamics ,11,20. AThe primary limitations of this study were as follows: first, the relatively small sample size and the difference in sex ratio and, second, the absence of other choroidal circulatory examinations. To further investigate choroidal changes in healthy participants, the relationship between choroidal function and morphology should be assessed in large and equally sexed samples using both LSFG and optical coherence tomography angiography. Future studies should compare the MBR of CSC eyes and healthy eyes using the cold pressor test to determine the relationship between the severity of CSC and the prediction of future onset.In conclusion, to the best of our knowledge, this is the first study to report changes in the MBR after a cold pressor test. Our data suggested that in young healthy individuals, as we hypothesized, an increase in sympathetic activity induced by a cold pressor test increases choroidal hemodynamics in the macula along with an increase in systemic circulatory dynamics, which normalizes after 10 min. These novel findings suggest that LSFG may provide a novel approach for assessing sympathetic activity and intrinsic vascular responsiveness in the eye. In addition, a comparison of the choroidal circulation in CSC eyes and healthy eyes using the cold pressor test may reveal the relationship between the severity of CSC and the prediction of future development.


Akkermansia, a relevant mucin degrader from the vertebrate gut microbiota, is a member of the deeply branched Verrucomicrobiota, as well as the only known member of this phylum to be described as inhabitants of the gut. Only a few Akkermansia species have been officially described so far, although there is genomic evidence addressing the existence of more species-level variants for this genus. This niche specialization makes Akkermansia an interesting model for studying the evolution of microorganisms to their adaptation to the gastrointestinal tract environment, including which kind of functions were gained when the Akkermansia genus originated or how the evolutionary pressure functions over those genes. In order to gain more insight into Akkermansia adaptations to the gastrointestinal tract niche, we performed a phylogenomic analysis of 367 high-quality Akkermansia isolates and metagenome-assembled genomes, in addition to other members of Verrucomicrobiota. This work was focused on three aspects: the definition of Akkermansia genomic species clusters and the calculation and functional characterization of the pangenome for the most represented species; the evolutionary relationship between Akkermansia and their closest relatives from Verrucomicrobiota, defining the gene families which were gained or lost during the emergence of the last Akkermansia common ancestor (LAkkCA) and; the evaluation of the evolutionary pressure metrics for each relevant gene family of main Akkermansia species. This analysis found 25 Akkermansia genomic species clusters distributed in two main clades, divergent from their non-Akkermansia relatives. Pangenome analyses suggest that Akkermansia species have open pangenomes, and the gene gain/loss model indicates that genes associated with mucin degradation (both glycoside hydrolases and peptidases), (micro)aerobic metabolism, surface interaction, and adhesion were part of LAkkCA. Specifically, mucin degradation is a very ancestral innovation involved in the origin of Akkermansia. Horizontal gene transfer detection suggests that Akkermansia could receive genes mostly from unknown sources or from other Gram-negative gut bacteria. Evolutionary metrics suggest that Akkemansia species evolved differently, and even some conserved genes suffered different evolutionary pressures among clades. These results suggest a complex evolutionary landscape of the genus and indicate that mucin degradation could be an essential feature in Akkermansia evolution as a symbiotic species. Due to ulations . In addi disease . Furthertbiotics .Akkermansia is an interesting case of a member of the GI microbiota with close relatives inhabiting non-related environments. Furthermore, Akkermansia-related species are almost the only members of the Verrucomicrobiota phylum identified in the vertebrate gut microbiome, especially in mammals , calculating the pangenome of different Akkermansia genomic species, searching for genes that could be involved in the differentiation of this genus, and calculating the evolutionary pressure of several genes from the most represented genomic species.In this study, in order to understand the (pan)genomic transformations involved in the divergence of a gut colonizer from free-living relatives, we performed a phylogenomic analysis of a set of more than three hundred 2.2.1.classify_wf’ of the GTDB-TK program, version 2.1.0 (Akkermansia genus (“g__Akkermansia”). Genome completeness and contamination were calculated using the program ‘lineage_wf’ from CheckM version 1.1.3 (A set of six hundred candidate genomes classified the Akkermansiaceae family was downloaded from the NCBI Genbank FTP site (May 2022). Those genomes were evaluated using two criteria: their taxonomic identity and their degree of completeness and contamination. The taxonomic identity was confirmed by using the program ‘on 2.1.0 , using ton 1.1.3 ; only thon 1.1.3 .2.2.de novo using Prokka, version 1.11 (--metagenome --kingdom Bacteria --addgenes). Orthogroups from the set of the final dataset were calculated by Orthofinder version 2.5.5   to generate a maximum likelihood-based tree with an aLRT with 1,000 replicates as the branch support test. The phylogenomic tree was visualized using the Toytree Python package  ; the alion 2.1.4   with the prediction of clusters defined by average nucleotide identity (ANI) values. All genomes were compared in an all-vs-all manner using FastANI version 1.32 . The orthogroup matrix (including unassigned orthogroups) was obtained for each run and utilized for different pangenome metrics. Pangenome curves were created using the panplots function in R (created by SioStef)curve_fit from scipy python package, using the equation “y = 𝛾,”ax applied on the panplots output. We calculated the shell, cloud, “soft-core” and core components of the pangenome from the complete orthogroup matrix by using Python scripts with the pandas package, considering the following criteria: core gene families as the orthogroups present in 100% of the strains, soft-core groups as present in between 99.999 and 90% of the strains, shell as groups present between 89 and 15% of strains, and cloud as the gene families present in between 14% and the equivalent to two strains. Unique groups can be deduced from the set of “species-specific orthogroups,” and the “unassigned genes,” both reported by Orthofinder. Figures were created with ggplot2 and the ggarrangment packages.In order to analyze the pangenome of the main 2.5.Akkermansia proteomes were analyzed using EggNOG mapper version 2.1.6 .on 2.1.6   of each species cluster (or the current proteome for 1-genome species groups) were compared by using Orthofinder as previously mentioned. The generated orthogroup matrix was utilized for the generation of a phylogenomic tree, as previously mentioned, confirming that the position of the species in the tree was the same as the one observed in the previous tree. This tree and the binary version of the complete orthogroup matrix were used by the software Count . Alignments were converted into codon-aware aligned using PAL2NAL  were inferred using HGTector v2.0b3 . Selecte2.9.shapiro.test). We compared data groups from two conditions with non-normal distribution using the Mann–Whitney U test (wilcox.test).All statistical tests were conducted in R version 4.1.0. Normal distribution was evaluated using the Shapiro–Wilk test , could not be included in the final dataset due to their below-cutoff completeness . Most species clusters were previously detected in the GTDB-TK reference. The four most prevalent clusters represented Akkermansia muciniphila, Akkermansia sp004167605, Akkermansia muciniphila_B, and Akkermansia sp001580195. Each cluster contained 193, 73, 40, and 19 genomes, respectively . Compared with previous studies . Unless specified, we used this 367-genome dataset (and its corresponding genome species clusters) for all downstream analyses.A representative tree was created from the original dataset , includi studies , the GTD3.2.A. muciniphila pangenome studies have been performed multiple times .Recently, comparative genomics and le times . HoweverAkkermansia sp004167605) is the longest among the analyzed species, and Cluster 1 (A. muciniphila) contained the smallest core genome. The pangenome comprised between 6,865 and 3,515 orthogroups among those four species; these changes in pangenome content seem to depend on the number of genomes in each cluster, a feature seen in other open pangenome models  and information transfer  were the most prevalent in the core and soft-core fractions. On the other hand, categories such as X (“Mobilome”) or Q (“Secondary metabolite transport and metabolism”) had the highest percentages detected in the shell cloud or unique gene fractions. Genes without COGs have strongly higher percentages in the fraction of unique genes (data not shown). Cluster 2 and Cluster 4 were the genomic species with the highest number of core-enriched categories compared to the accessory genes. COG categories U, G, Q, P, C, D, N, F, H, and E showed high core-to-accessory ratios in Cluster 2, while Cluster 4 showed I, K, C, E, Q, F, H, P, J, and N presenting the largest ratios. The categories that overlap these two Clusters correspond to P, C, F, H, and N. Cluster 1 showed that category J was core-enriched, and Cluster 3 showed that P and E categories were core-enriched –D. Categ3.3.Akkermansia genome dataset utilized in this study represented 25 potential genomic species , in comparison with a set of members of Rubritalea, Roseibacillus, Luteolibacter and Haloferula, the closest Verrucomicrobiales as outgroups . This feature suggests that Lineage A was more conservative than Lineage B in its gene content. In general, gene acquirements were most frequent than gene loss events across the Akkermansia phylogeny.According to the prediction, ancestor . The preancestor  experienLAkkCA  category  and the production of propionyl-CoA via propionyl CoA:succinate CoA transferase (COG0427). The possession of NADH dehydrogenase, ATPase, and some terminal oxidases suggest an inheritance of the respiratory metabolism from their non-Akkermansia ancestors, something noticeable if we consider that previous studies suggested that A. muciniphila may conduct microaerobic metabolism in the mucus layer niche , in addition to a member of COG1225 (Bcp Peroxiredoxin), COG0450 (AhpC peroxiredoxin), COG1592 (Rubreythrin), COG0605 (Superoxide dismutase) and COG0753 . This latter finding suggests the need for a proper adaptation to higher oxygen levels during the origin of the Akkermansia genus.The prediction of the putative genetic content that harbored the hypothetical last common ancestor of this complete genus raises the opportunity to predict the properties of early members of the f LAkkCA  includesf LAkkCA ], and ther niche , even ifLAkkCA also contained a series of proteins involved in its relationship with the host and environment. For example, this ancestor contained a set of genes encoding proteins containing COG0666 , a ubiquitous domain involved in protein–protein interactions. In Bacteria, ANKRs were mostly investigated in proteobacterial organisms, especially pathogens, where those proteins could be associated with protein secretion systems involved in pathogenic interactions with the host  of the same protein across the outer membrane of Gram-negative bacteria . In this ancestor, we also found another CAZyme family called GH16_3. This GH family comprises O-glycanase capable of targeting the polyLacNAc structures (consists of repeated N-acetyllactosamine units) within oligosaccharide side chains of both animal and human mucins , GH84 (N-acetyl β-glucosaminidase), GH89 (α-N- acetylglucosaminidase), and GH110 , were also found in Akkermansia species as expected , A24A (type 4 prepilin peptidase), and S41A , for example, seem to have housekeeping functions. In contrast, some families such as U32 (collagenase), I04 , or M42 , seem to be acquired as special adaptations. For example, bacterial collagenases have been associated with pathogenic bacteria , followed by members from the Proteobacteria and Bacteroidota phyla. Interestingly, we detected in the clusters of Akkermansia spp. different genes associated with families of glycosyl hydrolase enzymes , with a predicted donor from Bacteria (superkingdom) and Bacteroidota. Some of these enzymes were found in intestinal bacteria such as Bacteroides plebeius, Bifidobacterium longum subsp. infantis and Fibrobacter spp. , we calculated the Tajima D values for the core gene families for each species. Our analysis (Akkermansia sp004167605) showed a shift to more negative values. Cluster 1 (corresponding to the bonafide A. muciniphila) showed a more extended distribution, as seen as well in the boxplots , with a common language effect size (CLES) value of 0.809, which means that 80.9% probability that randomly chosen observation from one group will be greater than a randomly selected from the other group, making this difference substantial. In the same way, in comparison between cluster #1 and cluster #3 and between cluster #1 and cluster #4, both comparisons showed significant differences . However, CLES values show that both comparisons suggest that differences are more modest . Based on the aforementioned information, our results show that the differences between clusters are sufficient to back up the potential description of new Akkermansia species. Differences between cluster #2 versus cluster #3, and cluster #2 versus cluster #4 were also significantly different, although with lower CLES values . This information suggests that the more negative distribution in Cluster 2 is significant in relation to the other three clusters.Tajima D statistical test is a population genetic test used to elucidate if a gene family evolved in a neutral manner, or if it evolved under a non-random process, such as balancing selection, demographic expansion, or contraction, among other effects . In ordeanalysis  showed tanalysis , which Cboxplots . In addip-value) may reflect the effect of positive selection since it indicates an excess of rare alleles, suggesting population expansion. In counterpart, very positive D values (above 2) suggest a high proportion of common alleles, indicating balancing selection or population contraction  were annotated with EggNOG mapper, a posterior approach was the classification of Tajima value distribution across different COGs categories based on each cluster . It is p3.8.Akkermansia species represented in the genome dataset raise the possibility that conserved genes among different species groups have different selective pressures. In order to elucidate that aspect, the pairwise dN/dS ratio was compared from the core, single-copy, orthogroups between a symmetrical amount of genomes (n = 19) from those mentioned above the top four Akkermansia genomic species. Since dN/dS ratios are metric values for macroevolution, all comparisons were contrasted using the genomic content from A. glycaniphila as the outgroup; this ensured that all comparisons were compared with each other using representatives from the same outgroup. Additionally, all dN/dS comparison datasets between representative genomes  were compared using the Mann–Whitney U test, discarding all groups without significant differences.Different gene diversities among the main Akkermansia species , there are only a few studies focused on the evolutionary dynamics of gene gain/loss events . Moreovphibians , it is rtebrates . Moreovetebrates . Mucins,tebrates , are impntestine  and elicntestine , among ontestine . Our stuBacteria domain (through HGT), opens the way to the possibility that the genus Akkermansia obtained genes  from various intestinal taxa, or they were created by Akkermansia spp. and have been transferred to other microorganisms, or even that genes transferred to Akkermansia have been perfected through contact with various intestinal microenvironments of different hosts, to be later transferred to different taxa.In addition, the fact that most of the genes were cataloged in the Akkermansia, as well as, the existence of a copious taxonomic diversity for this genus. A systematic phylogenetic analysis must be implemented to decode the probable taxonomic origin of several HGT-derived genes.Our results show the necessity of new studies with the goal to understand the unique new species and the intraspecific evolutionary processes of Akkermansia isolates from several other members of the vertebrates may help to gain more insight into the relationship between mucin, the evolution of vertebrates, and the natural history of Akkermansia. Moreover, the understanding of the evolutionary process involved in the origin of the machinery to process mucin in Akkermansia spp. could help to understand the relationship with the maintenance of gut health, as well as their improvement through the design of probiotics.Finally, the need to explore and discover more The original contributions presented in the study are included in the article/JC conceived and directed the study. DG, MM-O, BV-V, and JC analyzed the data. All authors collaboratively elaborated, edited, corrected the text and figures in the manuscript, read the manuscript, and approved the content.ANID Doctorado Nacional/2021-21211564.JC was supported by ANID Fondecyt Project #11200209. BV-V was supported by The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


A fibre supplement containing glucomannan, inulin, psyllium, and apple fibre was the most effective at reducing the BMI, body weight, and CRP (p = 0.018 for BMI and body weight and p = 0.034 for CRP compared to placebo at the end of the intervention). Overall, the results suggest that dietary fibre supplements in combination with ER may have additional effects on weight loss and the metabolic profile. Therefore, taking dietary fibre supplements may be a feasible approach to improve weight and metabolic health in obese and overweight individuals.Obesity and its associated complications require various lifestyle changes and treatment options. Dietary supplements are considered an attractive alternative to traditional therapy, mainly because they are accessible to the general population. The aim of this study was to investigate the additive effects of a combination of energy restriction (ER) and four dietary supplements on changes in the anthropometric and biochemical parameters in 100 overweight or obese participants who were randomly assigned one of the dietary fibre supplements containing different dietary fibres or a placebo for 8 weeks. The results confirmed that fibre supplements plus ER significantly ( Obesity has reached the epidemic level across the world, and its prevalence continues to increase . MoreoveAmong the various alternatives for weight control, the use of dietary supplements from natural sources has become popular in recent years. A recent systematic review and meta-analysis of 62 human studies showed that viscous fibre intake affects body weight, body mass index (BMI), and waist circumference independent of an energy-restricted diet . There iAmorphophallus konjac [Common nondigestible carbohydrates (NDCs) in dietary supplements are glucomannan, inulin, psyllium, and β-glucans. Glucomannan is a natural dietary fibre composed of β 1,4-linked D-mannose and D-glucose monomers derived from a tuber called s konjac . Its mecs konjac . Inulin s konjac ,21. Howes konjac . β-glucas konjac . Psyllius konjac . Psyllius konjac , a feelis konjac , the secs konjac , and by s konjac . Differes konjac .On the market, numerous dietary fibre supplements with different formulations are available. Because of the conflicting results regarding the nature of dietary fibre supplements for weight loss and the attenuation of certain obesity-related metabolic diseases, the objective of the present study was to investigate the influence of four commercially available dietary fibre supplements on the anthropometric and biochemical parameters in a population of overweight and obese patients who also had dyslipidaemia and mild hypertension. As energy restriction is the most accepted weight loss strategy, all supplementation interventions were performed in combination with energy restriction and compared to the energy restriction alone. Regardless of the putative beneficial health effects, the supplements will only be accepted by the consumer if they are also sensory-acceptable and do not cause gastrointestinal symptoms, and therefore the two parameters were followed concurrently. The hypothesis of our study is that dietary fibre supplements with energy restriction aid in weight loss and improve the lipid profile in overweight and obese participants.ClinicalTrials.gov (NCT05333315). All subjects signed written informed consent forms before the conduction of the study and the Declaration of Helsinki was followed.The study was a five-arm, parallel, randomized, control trial conducted at the University of Primorska, the Faculty of Health Sciences, between April and July 2022. The study protocol was approved by the Slovenian National Medical Ethics Committee  and registered at The participants were randomly allocated to one of five groups: the investigation product (IP1–IP4) groups or the placebo product (PP) group, before the initiation of an 8-week energy-restricted weight loss period. Stratified variables were gender and age. Blinding was not possible because some products were capsules, liquids, powder, or soft chews, and therefore the experiment was performed as an open-labelled experiment. The participants attended screening visits before the intervention period and on three clinical investigation days: at baseline (week 0), at the end of the first month of the intervention (week 4), and at the end of the whole intervention (week 8). At each time point, the participants had a consultation with a dietitian, including an evaluation of diet and body composition, blood pressure was measured, and blood samples were collected.The primary outcome was the effect of dietary fibre supplements during energy restriction on body weight, compared with the placebo supplement. Secondary outcomes were the effects on body composition, blood pressure, gastrointestinal symptoms, and biochemical markers.2, no more than a 3% change in body mass within the last three months, age between 40 and 60 years, and a willingness to avoid the consumption of any food supplements at least 2 weeks before and during the study. The exclusion criteria included taking any prescribed medication or food supplement within the two weeks preceding the study; any clinically significant history of serious digestive tract, liver, kidney, cardiovascular, or haematological disease or diabetes; gastrointestinal disorders or other serious acute or chronic diseases. A study power calculation  was performed based on literature data [The study population included healthy adults (from 40 to 60 years old) recruited from the Primorska region of Slovenia. Volunteers were recruited through an advertisement posted on Internet forums, sent via e-mail lists, and published in local newspapers. Only healthy subjects were included. Additional inclusion criteria were a BMI between 25 and 30 kg/mure data . We calcOnce eligibility was confirmed, the following information was collected: age, gender, and ethnic origin of the subject; height (cm) and body weight (kg); body fat mass (kg); and visceral fat index. The participants did not receive any compensation for their participation in the study.The study was conducted with four different dietary fibre supplements (IP formulations) and a placebo product (PP) produced by Biostile d. o. o., Comen, Slovenia. The details on the content, form, and administration of the different dietary fibre supplements are shown in ® Microlife, Medical Home Solutions, Inc., Golden, CO, USA) was multiplied by the appropriate factor of physical activity (from 1.3 to 1.6), and then a reduction of 837 kJ/200 kcal–2093 kJ/500 kcal was made. The macronutrient composition of the diet was approximately 48–52% EI of carbohydrates, 30–33% EI of fat, and 18–22% EI of protein. Dietary intake of the participants was tracked during the study using a 24 h recall. An expert dietitian carefully collected a list of all foods and beverages, using food models. Dietary data were analysed using the Open Platform for Clinical Nutrition (OPEN), which can be accessed through the website http://opkp.si/ (accessed on 2 October 2022).At the beginning of the study, an energy-restriction plan was prepared for each participant. After nutritional data collection at baseline, all subjects participated in two educational sessions (2 h) on healthy diet, nutrient composition, the proper timing of eating, and the beneficial effects of the daily consumption of vegetables and fruits. All subjects also attended two sessions of individual consultations, during which they were given a personalized diet plan for eight weeks. To estimate the total energy needs, an individual’s RMR measured with an indirect calorimeter . BMI was calculated as weight (kg) divided by height (m) squared. Body composition ) was assessed using bioelectrical impedance analysis (BIA) Tanita MC-980MA  and dedicated software (GMON Pro-Tanita).Additionally, blood pressure was measured with an automatic device  after 10 min of rest in the supine position. The measurement was repeated twice.g, frozen, and stored at −80 °C until subsequent analysis. Serum concentrations of glucose, TAGs, total cholesterol, LDL cholesterol, HDL cholesterol, total bilirubin, AST, ALT, total bilirubin, uric acid, and C-reactive protein (CRP) were measured with a Cobass c111 analyser (Roche). Serum concentrations of interleukin-6 (IL-6), tumour necrosis factor alpha (TNF-α), lipopolysaccharide binding protein (LBP), and zonulin were determined in duplicate on a microplate reader  using human ELISA Kits . Assay sensitivities were 0.32 pg/mL for IL-6, 0.13 pg/mL for TNF-α, 0.13 ng/mL for LBP, and 0.5 ng/mL for zonulin. Assay interassay and intraassay CVs were typically between 5% and 12%.Venous blood samples were collected in 6 mL vacuum test tubes  after a 12 h overnight fast (complete dietary restriction with the exception of water). Serum was immediately separated by a 10 min centrifugation at 2000× During the fourth week of the intervention, the participants were asked to evaluate the products. The participants were provided a prescribed questionnaire that contained three items: the overall impression and likeability of the product, and the acceptability of its taste and its texture. The evaluation scores were from 1 (dislike very much) to 10 (like very much) for the likeability of the product, and from 1 (dislike very much) to 5 (like very much) for taste and texture, separately.Moreover, participants were also asked to report how likely they were to continue consuming the product, to report the possible gastrointestinal discomforts, and to self-report changes in appetite while consuming the product. Gastrointestinal discomforts, self-reported changes in appetite, concomitant medication, and adverse events were registered on the clinical investigation days and during the consultations with the dieticians.t-test or Wilcoxon signed-rank test, whereas the comparison of the mean changes between groups was analysed using an independent t-test or the Mann–Whitney U test. Moreover, to compare the means at baseline or the change at week 4 or 8 weeks from baseline between the groups, the one-way ANOVA was performed, and p-values < 0.05 were considered statistically significant.Statistical analysis was performed using SPSS version 23.0 . The statistical analysis was performed in two ways: an intention-to-treat analysis was performed on age, body weight, BMI, and fat mass, in which all participants were included, and a per protocol analysis of the treatment efficacy, which was performed on all participants who completed the 8-week intervention. All results are presented as the mean value with standard deviation (SD). Normality of variables was tested using the Shapiro–Wilk test. The effects of the interventions within each group were analysed with a Student’s paired-samples n = 1; IP3, n = 2; IP4, n = 1; PP, n = 2), ten participants withdrew due to adverse symptoms , and four participants were excluded because of protocol violation . The flowchart of the study is illustrated in Overall, 100 participants were included in the study. There were 83 females and 17 males, evenly distributed across the five treatment groups. In total, 80 of the 100 participants (20% dropout) completed the 8-week study. Six participants were COVID-19-positive during the study  randomised to five groups. At baseline, there were no significant differences (p > 0.05) in age, body weight, BMI, or fat mass across the five groups. If we consider only the participants who completed the study, then the BMI was significantly lower in the PP group. However, the body weight, percentages of fat mass and fat-free mass, and visceral fat rating did not significantly differ between the groups (Baseline data  show thee groups .Adherence to the treatment (monitored by the investigator by counting the medication bags returned by patients at each follow-up visit) and adherence to diet (assessed by patients at each follow-up visit) were generally high in all groups . No statistically significant differences were found between the groups in the adherence or the self-assessment of the level of adherence to the diet. Overall, the self-reported compliance was 89% consumption of the intended intervention products, with 92% in the IP1 group, 87% in the IP2 group, 86% in the IP3 group, 93% in the IP4 group, and 90% in the PP group.p < 0.01) reduced body weight and BMI 4 and 8 weeks after the start of the study, while in the PP group, the significant differences in body weight and BMI were observed only after 8 weeks of energy restriction (p = 0.002 for BMI and body weight), IP3 and PP (p = 0.050 for body weight and p = 0.029 for BMI), and IP4 and PP (p = 0.027 for body weight and p = 0.028 for BMI) groups after 4 weeks of intervention. Moreover, the decrease after 4 weeks of supplementation was significantly higher in the IP2 group than in the IP1 group (p = 0.041 for body weight and p = 0.021 for BMI) and IP3 group (p = 0.048 for body weight). After 8 weeks of the intervention, reductions in body weight and BMI were significantly different only between PP and IP2 (p = 0.018 for BMI and body weight) and between PP and IP4 for BMI (p = 0.049).The first objective was to test whether dietary fibre supplements combined with energy restriction are able to reduce body weight and BMI. The result confirmed that the administration of IP1, IP2, IP3, and IP4 significantly (triction . The stup < 0.01) in groups IP1, IP2, IP3, and IP4, and this reduction was even greater after 8 weeks (p < 0.01) compared to the baseline measurement. However, the reduction in fat mass and visceral fat was also significant in the PP group after 8 weeks of energy restriction. The study showed that the reduction in visceral fat was significantly different only between PP and IP2 (p = 0.019), but only after 4 weeks of the intervention. However, the FFM was also significantly affected in all groups (except PP). A significantly higher reduction was observed in the IP2 group than in the PP group (p = 0.021) and in the IP1 group than in the PP group (p = 0.032) after 4 weeks of intervention, and even after 8 weeks of intervention . Overall, the IP2 supplementation was the most effective at reducing not only body weight, BMI, and visceral fat, but also FFM  in groups IP1 and IP4, while DBP decreased significantly in groups IP2 and IP4 after 4 weeks (IP2) and 8 weeks (IP2 and IP4) compared with the baseline measurement (p = 0.049 at 4 weeks and p = 0.050 at the end of the intervention). No other differences were found in relation to blood pressure ; at week 8, the differences were no longer significant  (p = 0.046)  . The ser= 0.034) . Due to = 0.034) . The red= 0.046) .Participants were asked to rate how much they liked the product in general, and how acceptable it was in terms of taste and texture . The meaAs shown in Because energy restriction was prescribed to the participants of all groups, dietary intakes were analysed for the baseline and for week 8. Even though the energy intake decreased in all the groups and the macronutrient composition of the diets changed due to the dietary recommendations, neither the energy intake nor the macronutrient composition of the dietary intake differed between the five groups . This alMany studies have found that increasing fibre intake results in increased satiety, reduced hunger, decreased energy intake, and weight loss ,34. The The influence of dietary fibre supplements on energy regulation remains controversial . TherefoIn addition, a similar combination (inulin (nonviscous fibre) and β-glucans (viscous fibre)) in the IP4 group also significantly reduced body weight and BMI at 4 weeks and BMI at 8 weeks in comparison to the placebo. These results are very interesting, as the amount of fibre in this supplement was very low compared to the literature; nonetheless, this product showed very good results. It has been reported that the ingestion of β-glucans in an amount ≥ 3 g causes a decrease in body weight in people with type 2 diabetes for at least 3 weeks ,40. The Overall, IP2 and IP4 were the most effective supplements in reducing body weight (−5 kg for IP2 and −4 kg for IP4 in 8 weeks) and BMI. It is known that even slight reductions in weight can produce metabolic improvements. Each kilogram of lost body weight has been connected with a 16% reduction in type 2 diabetes risk . NeverthThe present study and others  report tw/w) of fibre treatment  beneficially enhanced the mucosal barrier function and anti-inflammatory status in mice [There is no consensus in the literature on the anti-inflammatory effects of dietary fibre. The results of two epidemiological studies suggest that the consumption of dietary fibre is inversely associated with serum CRP concentrations ,57, but  in mice . Our resIn this study, we also investigated whether 8 weeks of increased fibre intake would influence the participants’ gastrointestinal comfort, whether the increased fibre intake was acceptable for the participants, and whether the consumption of dietary fibre supplements led to decreased appetite. After 4 weeks of dietary fibre supplementation, the data indicated an increase in the frequency of bloating and a hardening of the stool consistency in all four conditions of dietary fibre supplementation. There was no evidence that other side effects, such as bloating or cramping, increased under fibre supplementation. This suggests that, on the whole, the participants did not experience significant adverse gastrointestinal reactions due to the increased fibre intake. However, there was some abdominal discomfort and flatulence, which is in line with previous studies that report the positive associations between fibre intake and increased flatulence, bloating, uncomfortable abdominal distention, and changed stool consistency ,62. MoreThe results of this study were limited by a relatively small sample size, which may explain the lack of significant results when comparing between groups. In addition, meals and the consumption of the test products were self-administered; thus, the possibility of noncompliance could not be excluded. Misreporting of intake may have occurred in the current study.In conclusion, based on the improvements in the health parameters observed in the present study, and because of the extensive dietary fibre intake gap between the recommended intake and actual dietary intake in Slovenia , fibre s


Steel slag is a solid waste produced in crude steel smelting, and a typical management option is stockpiling in slag disposal yards. Over the years, the massive production of steel slags and the continuous use of residue yards have led to vast occupation of land resources and caused severe environmental concerns. Steel slag particles can potentially be used as aggregates in concrete production. However, the volume stability of steel slag is poor, and the direct use of untreated steel slag aggregate (SSA) may cause cracking and spalling of concrete. The present research summarizes, analyzes, and compares the chemical, physical, and mechanical properties of steel slags. The mechanism and treatment methods of volume expansion are introduced, and the advantages, disadvantages, and applicable targets of these methods are discussed. Then, the latest research progress of steel slag aggregate concrete (SSAC) is reviewed. Using SSA leads to an increase in the density of concrete and a decrease in workability, but the mechanical properties and durability of SSAC are superior to natural aggregate concrete (NAC). Finally, future research in this field is proposed to motivate further studies and guide decision-making. Concrete is the most-used building material globally, with global consumption of about 12 billion tons annually [Steel slag is the byproduct of the crude steel smelting process, and the discharge is approximately 12–20% of crude steel production . The ann2S, C3S, C4AF, and C3A in steel slags are typical mineral phases of Portland cement, indicating the potential of steel slags to function as hydraulic materials. Therefore, one recycling option is to grind steel slags into a fine powder and use it as a supplementary cementitious material. It has been proved that replacing cement with 10–20% steel slag powder does not weaken the mechanical strength of the system [Steel slag could be valuable for promoting a sustainable relationship between the concrete and steel industries. Ce system ,10,11, be system . Steel se system . GrindinAnother recycling option is to sieve steel slag into a good grading range, then use it as a coarse/fine aggregate to produce concrete. This option is the most likely option to realize the bulk utilization of steel slag, which is significant in alleviating the resource and environmental dilemma of the concrete and steel industries. However, the volume stability of steel slag aggregate (SSA) is poor, and the prepared concrete has the risk of cracking . To dateSlags of various types account for approximately 90% (by mass) of the byproducts discharged from the crude steel smelting process; the others are gas, dust, and sludge . Figure BOF steel slag is the solid waste produced in the converter steelmaking process, and its discharge is approximately 100–150 kg per ton of BOF steel ,21,22,23EAF steel slag is the byproduct of electric furnace steelmaking, the output of which is approximately 110–200 kg per ton of EAF steel ,23,26,27LF steel slag is the solid waste produced by the secondary refining of BOF and EAF steel. About 10–50 kg of LF steel slag is discharged for each ton of crude steel refined ,27,28,292, FexOy , Al2O3, and MgO is nearly 90%, while the rest covers the oxides of P, Mn, S, Ti, Na, K, Sr, and others. The chemical compositions of BOF and EAF steel slags are somewhat similar; despite this, the predominance of higher contents of specific components can be analyzed, due to their different steelmaking processes. BOF steel slag has a higher CaO content, while EAF steel slag has higher SiO2, FexOy, and Al2O3 concentrations. Furthermore, oxides of Cr, Pb, V, Mn, Zn, and other heavy metals can be observed in steel slag [eel slag . In the eel slag ,32.2S), alite (C3S), brownmillerite (C4AF), wustite (FeO), and srebrodolskite (C2F). The most common mineral phases in EAF steel slag are larnite (C2S), wustite (FeO), magnetite (Fe3O4), gehlenite (C2AS), and quartz (SiO2). The C2S, C3S, and C4AF crystal size in steel slags is larger than in cement, and the content is much lower [2S, wustite, gehlenite, and CaO-FeO-MnO-MgO solid solution, cannot form new products in the hydration process, and they only participate as fillers [3S, and C4AF can react with water and show less hydration activity [3), portlandite (Ca(OH)2), and brucite (Mg(OH)2) have also been reported in the previous literature, and these mineral phases are mainly formed during the aging process of steel slag [There are more than ten kinds of mineral phases in steel slag that can be observed. The most common mineral phases in BOF steel slag are larnite 2 produced by the hydration of cement, which has a regular layered or sheet-like structure [2 crystals causes the concrete to crack, providing enough room for ettringite growth. The growth of ettringite further promotes the expansion of cracks, allowing more water to enter the concrete, and further aggravates the expansion of steel slag. The continuous generation of Ca(OH)2 and ettringite crystals eventually cause cracking and spalling of the concrete.The aggregate is tightly embedded in concrete, with little room for free expansion. The local stress caused by the expansion of SSA can crack concrete easily, or the concrete structure could even suddenly burst during service. The specific crack morphology is shown in tructure . SimultaSteel slag produced by steel mills must be preliminarily treated for a high cooling rate. Common preliminary treatment methods include water spray, water quenching, air quenching, and instantaneous slag chill . These mNatural aging is a traditional post-treatment option for steel slag stabilization. The steel slag discharged from the steel mill is stored in the slag disposal yards and relies on rain and moisture in the air to consume f-CaO and f-MgO. Natural aging does consume a certain amount of f-CaO and f-MgO in the early stage, but it also leads to structural densification of the steel slag surface and prevents further reaction ,40. ChenTherefore, natural aging is slow and time-consuming, occupying a large area of slag disposal yards to provide adequate stockpiling area ,120.Several techniques have been developed based on the same principle to solve the problems of prolonged time consumption, low efficiency, and poor modification quality of natural aging. Kumar et al.  proposed2 concentration conditions, CaO, MgO, and other calcium-containing minerals in steel slag rapidly form stable carbonate crystals, called accelerated carbonation [2 in steel slag, effectively turning steel slag into a safe and stable carbon sink. According to different reaction conditions, the accelerated carbonation of steel slag can be divided into dry and aqueous.Under high-temperature, high-pressure, and high-CObonation ,124,125.2 gas in a high-temperature and -pressure environment. The chemical processes of dry carbonation are presented in Equations (1)–(3), and the technical flow is shown in 2 gas diffused into the steel slag through the vesicular pore structure. Then, the solid chemical components such as CaO, MgO, and Ca(OH)2 are carbonated to produce CaCO3 and MgCO3. The dry carbonation reaction can directly feed flue gas from steelmaking into the reactor to simultaneously achieve the capture of CO2 and stabilization of steel slag [Dry carbonation refers to the reaction of solid steel slag with COeel slag .CaO(s) 2S, and C3S in steel slag are leached and hydrated, and the hydration products are Ca(OH)2 and C-S-H. The chemical processes involved in the process are shown in Formulas (4)–(6) [2S and C3S and hydration products Ca(OH)2 and C-S-H. Subsequently, CO2 gas is dissolved into water to form CO32−, and CO32− reacts with the above active components to form C-S-H and CaCO3. The C-S-H is an intermediate product, which will further carbonize to form SiO2 and CaCO3 [Compared with dry carbonation reaction, aqueous carbonation is a complex three-phase reaction process, which can obtain a higher reaction rate and carbonation level at a relatively low reaction temperature (still higher than room temperature) and pressure conditions ,129. The (4)–(6) ,41,43,51nd CaCO3 . The chend CaCO3 ,43.CaO 2 [2 [3 produced by accelerated carbonation are aragonite (dry carbonation) and calcite (wet carbonation), and their crystal hardness reaches 67 GPa and 72.8 GPa, respectively. These mineral phases can form a dense organizational structure in the steel slag [The particle size of steel slag is an important factor affecting the accelerated carbon rate and the treated quality. On the one hand, as particle size increases, the specific surface area of particles decreases correspondingly, making steel slag less reactive to CO2 . On the 2 [2 . Therefo2 [2 ,125,128.2 [2 ,133, as eel slag . TherefoJiang et al.  investigThe principle of molten slag converting is adding modifiers in remelted steel slag to convert its composition. Modifiers react with f-CaO and f-MgO in molten steel slag to crystallize f-CaO and f-MgO, thus improving the volume stability of steel slag. Long et al. added 5%, 7%, and 14% fly ash to molten steel slag as the modifier. After the reaction at 1580 °C for 30 min, the corresponding digestion rate of f-CaO was 56.99%, 63.69%, and 68.55%, respectively . Zhang eThe external restraint technique refers to confining SSAC with a steel/FRP tube to limit its cracking and spalling ,141. As 2 concentration, reaction time, and steel slag particle size, the CO2 capture capacity of steel slag per kilogram can reach 4.7–516.9 g [2 gas annually worldwide [2 sequestration is small.Accelerated carbonation is a highly environmentally beneficial technique that can achieve the desired modification quality in a short period. By optimizing the temperature, pressure, humidity, CO–516.9 g ,51,128. orldwide , therebyorldwide . TherefoMolten slag converting technology can consume f-CaO and f-MaO quickly and improve the hydraulic and pozzolanic activity of steel slag, significantly improving the grindability of the treated steel slag . HoweverFilling SSAC into steel/FRP tubes is an effective and easy application that could accept the uncertain expansion of SSA. In particular, there are no additional processing and usage limits for SSAs, which would be more attractive for construction units. The use of steel/FRP tubes may cause an increase in material costs. However, the initial cost may be balanced, or even outbalanced, by the reduction in maintenance cost throughout the service life due to the excellent corrosion resistance of the jacket . BesidesOverall, the introduced treatment methods can consume f-CaO during the total life cycle of steel slag. In the hot slag stage, hot steam aging equipment and molten slag converting equipment can be installed on the steel slag production line to treat the fresh steel slag according to its characteristics. For the steel slags stockpiling in slag disposal yards, f-CaO content and expansion risk should be tested first. Then, an assessment should focus on the proposal for further treatment. Steel slags with qualified volume stability can be directly used in the production of SSAC. For the steel slag that still has the threat of uncertain expansion, it can be considered to make carbonized artificial SSA or used for steel/FRP tube-confined SSAC composite structures. In this process, combining multiple technologies should be tried to balance the capacity and cost of steel slag modification. Research on pilot and industrial trials to connect these methods as a new complete system needs to be further conducted.Over the years, uncontrolled concrete production has led to the overexploitation of high-quality river sand and gravel, leading to severe concerns about resource depletion. Utilizing treated steel slag as coarse/fine aggregate in concrete production is a high-value solution that can alleviate the worry about resource depletion and solve the negative impact of steel slag stockpiling in slag disposal yards.cD refers to the density of concrete prepared with coarse SSA, fD refers to the density of concrete prepared with fine SSA, and r stands for the replacement ratio of SSA. The R2 of Formulas (10) and (11) are 0.49 and 0.68, respectively, caused by the difference in the concrete mix ratio and the density of selected steel slag in the literature.The density of SSA is 10–50% higher than that of NA. When designing the mix proportion, the amount of SSA should be calculated by the same volume, with the NA being replaced, but not of the same weight. 60Co in concrete increased by 6.6% and 14.5%, respectively, when SSA replaced 25% and 50% of coarse NA. Pomaro et al. observed that, by substituting 100% coarse SSA for NA, the half-value layer of 60Co and 137Cs increased 12~16.8% and 15.7~17.8%, respectively [In addition, the increase in density means SSAC could be potentially used as a shielding material against the nuclear radiation of X-rays, gamma rays, and neutrons. In work conducted by Baalamurugan , the linectively . In contectively . UtiliziBecause SSA has a higher water absorption than NA, using SSA in concrete reduces the amount of free water in the mix. In addition, the surface texture of SSA is rougher than NA due to its honeycomb pores and small “dust” on the surface, which increases the frictional resistance between particles during the flow of concrete paste. Therefore, the fluidity, cohesiveness, and water relativity of the SSAC mixture are generally poor. Using SSA can improve the mechanical strength of concrete. The improvement in mechanical strength of concrete can be attributed to the following aspects: (a) greater contact area between mortar and SSA. The vesicular nature and rough surface texture of SSA increase the contact area with the mortar see ; (b) A sIn addition, the elastic modulus of SSAC is higher than that of NAC. Pomaro et al.  reportedReplacing NA with SSA harms the water tightness of concrete. Substituting 50% to 100% coarse SSA for NA, the total water absorption could be maximized by 13% and 24% , and theCompared with the features of NAC, the frost resistance durability of SSAC is more satisfactory. On the one hand, the rough texture of SSA can activate a stronger interlocking effect in ITZ, resulting in a closer bond between SSA and cement mortar ,160. On 3 present on the surface of the SSA reaction with MgSO3 solution to form gypsum [Utilizing SSA has a beneficial effect in improving the chloride resistance of concrete. The conductivity of SSA is higher than that of NA, which reduces the overall resistivity of concrete and results in a higher total charge pass (TCP) of SSAC . Howeverm gypsum . Adding m gypsum .2 penetration of SSAC is 55% lower in natural carbonation and 40% lower in accelerated carbonation than NAC. Andrade et al. [2. Therefore, SSA has a lower carbonation depth in the same accelerated carbonation period than NAC.Concrete produced with SSA presents better carbonation resistance. Chatzopoulos et al.  reportede et al.  designede et al. ,161. Sim2 emissions per cubic meter [Although the volumetric expansion treatment brought additional costs, SSA is still highly economical as a sustainable building material. The price of SSA is only about 20% that of NA . Shen etic meter . In addiThe utilization of SSA in the concrete industry can effectively reduce the exploitation of high-quality river sand and gravel, thus alleviating growing concerns about resource depletion and environmental pollution. Simultaneously, the consumption of steel slag can release many occupied land resources and alleviate the conflict between the steel industry and land. In addition, some heavy metal oxides with significant biological toxicity exist in steel slag; Mn, Zn, Cr, Cd, Ni, Al, V, and Pb are examples that can be given . If steeIn the present article, the production, properties, and treatment method of SSA and the performance of SSAC were reviewed. Steel slag is the main solid waste of crude steel smelting, and the main chemical components are oxides of Ca, Fe, Si, Al, and Mg. The cementitious mineral in steel slag belongs to the crystalline phase, which has poor hydraulic activity but no pozzolanic activity. Steel slag particles have the characteristics of high-density, rough surface texture, and good mechanical properties, their specific gravity is 10–50% higher than that of NA, and their mechanical strength is comparable to basalt gravel and superior to granite and limestone gravel. Therefore, steel slag particles have the potential to be used as coarse/fine aggregate in concrete production.2S. Among them, f-CaO is the main reason for the volume expansion of steel slag. The harmful effects of using untreated steel slag directly as an aggregate are obvious and could cause cracking and spalling of concrete. Therefore, SSA must be treated to control the content of f-CaO. The methods used to modify the poor volume stability of SSA include natural aging, hot steam aging, accelerated carbonization, molten slag converting, and external constraint. These methods have made significant progress and have, partly, been put into practice. The advantages, disadvantages, and applicability of treatment methods in making steel slag treatment schemes should be taken into account, and different methods should be combined.Steel slag is still a solid waste rather than an available resource because of the poor volume stability of SSA. The expansion chemical components in steel slag include f-CaO, f-MgO, and β-CReplacing NA with SSA could increase concrete density. Thus, SSAC is a type of heavy-weight concrete potentially used as a shielding material for nuclear radiation of X-rays, gamma rays, and neutrons. Replacing 100% river sand or gravel with SSA can reduce the slump by 75% and 100%, respectively. In the design of concrete mix ratio, the amount of superplasticizer or filler (cemented or inert) should be adjusted according to the SSA type and content. Fine aggregate is not recommended to replace more than 50% ratio with fine SSA due to a serious deterioration in the workability of concrete, and the coarse SSA could replace NA at ratios up to 100%. Ongoing studies show that, thanks to ITZ enhancement and the superior mechanical properties of SSA, well-designed SSACs commonly show superior mechanical strength to NAC, and the compressive, flexural, and split tensile strength can be improved by up to 50%, 60%, and 60%, respectively.Using SSA in concrete can provide great economic, environmental, and sustainable development benefits. SSAC showed lower total lifecycle costs. The price of SSA is only 20% that of NA, which can significantly reduce concrete production costs. The water tightness of SSAC is poor, but it still has good resistance to freeze-thaw cycles, corrosion, and carbonation, which is an outstanding advantage of utilizing steel slags as coarse/fine aggregate without decreasing the durability of concrete. Using SSAC allows a longer service life of structures and saves maintenance costs during service life. In addition, the consumption of steel slag can reduce slag disposal yard occupation and environmental pollution risk and avoid over-exploitation of high-quality sand and gravel. Meanwhile, the acceptable leaching results indicate that SSAC showed adequate environmental safety. This case implies a perfect industrial-ecological relationship between the steel industry and the concrete industry for achieving sustainable development.(1)Combining artificial granulation and accelerated carbonization is a promising method for steel slag modification with good quality and high environmental benefits. However, this technique is still in the laboratory research stage. Future research should focus on simplifying the production process, increasing production capacity, and controlling production costs to realize the industrial production of carbonized artificial SSA.(2)Filling SSAC into steel/FRP tubes can limit the cracking and spalling of concrete caused by the expansion of SSA, and the expansion of SSA can reduce the negative impact of the interfacial gap effect in the service stage; such composite structures are promising. In future research, an assessment should focus on the mechanical properties and design model of steel/FRP tube-confined SSAC composite structures.(3)Different sources are inconsistent in reports on the performance of steel slag. Mix proportions and properties of concrete prepared with SSA from one source cannot be extended to all sources, which hindered the research and large-scale usage of SSA. Conducting a global data survey on the SSAC and creating a primary dataset is necessary for future research. Then, this primary dataset should be used to train an artificial neural network to predict the properties of SSAC.(4)The research on the mechanical properties of SSAC mainly focuses on cube compressive strength, flexural strength, and flexural strength. In the future, research should be carried out on the fracture mechanical properties and deformation ability of SSAC, and the bond properties between SSAC and steel bars, to provide data support for the application of SSAC in broader fields.(5)There is a gap in research on the heavy metal leaching behavior in SSAC, and works have only been carried out in the condition of a low, coarse SSA replacement ratio. Future research should be conducted for more complex situations, including different SSA types, different SSA particle sizes, and higher SSA contents, to guide the decision of the SSA utilization scheme.The analyses of existing works indicate that steel slag has the potential to be utilized as aggregates for concrete. However, many research gaps in this field need further study, thus providing more critical evidence for utilizing steel slag. On the basis of the literature review, the following recommendations for future research are forwarded:


T/dt ~ 109–1011 K/s) for the material, is a unique approach for the rapid “single-step” synthesis of nanomaterials and coatings. In this study, experiments were carried out with intertwined wire under a directional spraying device in atmospheric Ar ambience. Two load systems of Fe-Ni and Fe-Ni-Co were considered in this work. Electrical parameters and high-speed camera images were obtained to reveal the physical mechanism and dynamic process of explosive spraying. The morphologic and crystallographic results were characterized by SEM and XRD. The magnetic properties were measured via VSM equipment, and the parameters of saturation magnetization Ms, residual magnetization Mr, and coercivity Hc were emphasized in the hysteresis loop pattern. The experimental results indicate that a dense coating was prepared with extremely low porosity, and the morphology of the coating surface shows different regions characterized by solidified chunks and loose particles. XRD patterns showed that crystalline structures were discrepant under two load systems with different Ni weight proportions. Magnetic measurements gave a thin and narrow hysteresis loop, which represents loops with good soft magnetic properties. Quantitatively, coercivity Hc decreased from 59.3 to 52.6 and from 121.0 to 49.9 for the coatings not containing and containing Co under parallel and perpendicular fields, respectively.Fe-Ni-based nanocrystalline coatings with unique magnetic properties are widely used as soft magnetic materials and usually act as the core component in electronic devices. Nanocrystallized particles and thin films have become a popular contemporary research direction. Electrical explosion, characterized by an ultrafast atomization and quenching rate (d Soft magnetic material refers to the ability to obtain a large magnetization response under a small alternating magnetic field and is the core component in magnetic devices. Fe-Ni-based magnetic materials, typically as permalloys, are among of the most essential soft magnetic materials, characterized by relatively large anisotropic magnetoresistance (AMR) , high peTraditional bulk magnetic materials were always limited in their practical application, especially in high-frequency and microwave fields, because of their low resistivity. Since the rise of the nanomaterials’ field, attention has been drawn to the thin films or fine powders of magnetic materials. Later studies verified that nano-sized magnets (such as nanoparticles or nano-films) still reserve their outstanding properties and preserves the soft magnetic properties, while more attractive effects and satisfactory performance were revealed, such as the giant magnetoresistance (GMR) effect ,12, whicT/dt ~ 1011 K/s), quenching rate (~1010 K/s), as well as the intense mechanical effect (shock wave) during an electrical explosion make this method a powerful tool to prepare high-quality fine powders and spray coatings [Electrical explosion of wires (EEW) refers to when a pulse current passes through the conductor , rapid phase change occurs due to Joule heating, and the conductor undergoes liquefication, vaporization, and, finally, ionization into plasma. The extremely high temperature increase rate  and a Rogowski coil (bandwidth of 4 MHz), respectively. The resistive load voltage uR was estimated asLs refers to the inductance of the wire and its holder. The correctness of voltage and current measurement was examined by conducting the pulsed current with a low-inductance (nH level) ceramic resistor [Ed was calculated asThe discharge voltage resistor ,23. The High-speed cameras (Phantom VEO) were used to record time-resolved images of the explosion.Morphologic and crystallographic characterizations included SEM  and XRD , respectively. The XRD pattern of the sample was obtained using Cu Kα radiation (λ = 0.15406 nm) at the working voltage and current of 40 kV and 40 mA, and the recorded range at 2θ was from 30 to 80° with a step size of 0.02°. Magnetic performance was detected by the vibrating sample magnetometer (VSM-LakeShore 8604), and two magnetic field directions, namely, parallel and perpendicular to the coating surface, were applied to measure the magnetic properties of the samples. Furthermore, experiments were conducted in Ar gas ambience, and the prepared samples were stored in a vacuum chamber to avoid possible oxidation.Two experimental methods were used in this paper. System 1 involved an explosion of an intertwined wire made from Fe, Ni, and Co metal wires. System 2 involved an explosion of an intertwined wire made from Fe and Ni metal wires with different weight proportions. The coating composition was selected by changing the diameter of the Ni wire. In this paper, system 1 was mainly investigated with regard to electrical characteristics, dynamic process, and crystallographic results; meanwhile, system 2 was used as a comparison for the synthesized alloy coating. The detailed load parameters are listed in In the conditions of the joint explosion of multi-species metal wires, the electrical characteristics and dynamics behaviors are usually complex and display asynchronicity ,24. To pnλ = d2sinθ, and Scherrer’s law, D = kλ/Bcosθ, the crystal lattice distance d and grain size D can be calculated, where λ is the waveform of the Cu target (0.154 nm), k is the Scherrer constant (0.89), B is the FWHM of the diffraction peak, and θ is the degree of the diffraction peak. Detailed parameters are given in d and the calculated cell constant [Crystallographic characterization for all four samples was performed by XRD and is shown in constant . In addiMs saturates faster under the parallel field, indicating easy axis magnetization in the coating plane. To show the characteristics of coercivity Hc, partial enlarged patterns of magnetic field intensity from −1.0 to 1.0 kOe are presented as the insets in Hc is 59.3 and 121.0 Oe under the parallel and perpendicular field, respectively. When measuring sample 1, the Hc is the same, which is approximately 49.9 and 52.6 Oe under the two fields. Moreover, adding Co into the permalloy coating decreases the Hc, which is satisfactory for soft magnetic materials.The magnetic properties of the prepared alloy coating were measured via VSM at room temperature, as shown in Hc several tens to hundreds of Oe [Hc, which means the synthesized films by the EEW method are feasible and applicable. However, some studies indicate that the coercivity could reach less than 10 Oe, and the magnetic properties were influenced by numerous factors, such as film thickness, grain size, added elements, substrate materials, etc. [Coercivity is one of the most critical parameters that can represent the magnetic properties of soft magnetic materials. Common Fe-Ni-based magnetic film preparation methods, such as thermal evaporation, electrochemically deposited magnetron sputtering can synthesize film with the ds of Oe ,16,26. Ils, etc. ,28. DeepThe behaviors of the electrical explosion spraying of intertwined wire using a directional spraying device in atmospheric Ar ambience were investigated. Spatio-temporal images showed that the effective diameter of the funnel-like spraying area was, more or less, 3 cm. The morphology and crystalline structure of the two synthesized systems of Fe-Ni and Fe-Ni-Co coatings were measured. Dense coatings with a low porosity were prepared. For both systems, the morphologic characteristics showed discrepancies in different regions, manifested as solidified chunks in CR and loose particles in ER. XRD patterns indicated that the Ni weight proportion in the coating influenced the crystalline structure of the sample. Thin and narrow hysteresis loops were obtained for both systems, especially under the parallel magnetic field, which represented relatively good soft magnetic properties. The changes between parallel and perpendicular coercivity were 49.9 and 52.6 for sample 1 and 59.3 and 121 for sample 3, respectively. A nanocrystalline permalloy film with relatively good soft magnetic properties was successfully prepared via the method of directional electrical explosion spraying.