Peroneal split tears are an underrated cause of ankle pain. While magnetic resonance imaging (MRI) is useful for diagnosis, split tears are challenging to identify. The aim of the study was to investigate the association of peroneus brevis split rupture with abnormalities of the superior peroneal retinaculum (SPR), anterior talofibular ligament (ATFL), calcaneofibular ligament (CFL), joint effusion, morphology of the malleolar groove, presence of the bone marrow oedema and prominent peroneal tuberculum.n = 80) and one without . Two observers evaluated the soft tissue lesions, and three graded the bone lesions. Fisher’s exact test and Pearson correlation were used for analysis. The Bonferroni-Holm method (B-H) was used to adjust for multiple comparisons.Ankle MRI cases were assessed by independent observers retrospectively in two groups: one with peroneus brevis split tears (p < 0.05) more common in the split tear group after applying B-H. SPR total rupture was seen only in the experimental group. No differences in incidence of ATFL and CFL lesions or other SPR lesions were noticed (p < 0.05).Only bone marrow edema in the posterior part of the lateral malleolus was significantly (Bone marrow edema in the posterior part of the lateral malleolus is associated with peroneus split tears on MRI. Ankle disorders are a common clinical problem. Injuries to the ankle constitute up to 10% of all visits to the emergency room (ER)  of all iTo our best knowledge, no previous study regarding the relationship of the PB split rupture and lateral ligament injury has been conducted before.The pathophysiology of peroneus tendon split tears is not entirely understood . It has Clinical examination may be inaccurate in the acute situation due to pain . MRI or The International Olympics Committee (IOC) has repeatedly emphasized the importance of preventing injuries in the pathophysiology of peroneus split rupture . BecauseThe aim of this study was to investigate the association of peroneus brevis split rupture with abnormalities of the superior peroneal retinaculum (SPR), anterior talofibular ligament (ATFL), calcaneofibular ligament (CFL), joint effusion, morphology of the malleolar groove, presence of the bone marrow oedema and prominent peroneal tuberculum.2 in the experimental group and 23.7 in the control group (p > 0.05)  at the Sahlgrenska University Hospital (SU) in Gothenburg, Sweden, were eligible.MRI examinations of the adult patients (age > 18 years) performed between 2018 and 2021 (A dedicated ankle coil was used for MRI acquisition at 3.0 Tesla. Only MRI examinations with the following sequences were included: proton density (PD)-weighted with and without fat suppression, T2-weighted and T1-weighted without fat suppression.All ankle MRI examinations included PD-weighted turbo spin echo (TSE): echo time (TE) 45 ms and repetition time (TR) 2800–5000 ms; T2-weighted (TSE): TE 60 ms and TR 3000–5000 ms; and T1-weighted: TE 11.5 ms, TR 700–750 ms. Voxel size was 0.45 × 0.53 × 3.0 mm, slice thickness 3 mm and field of view (FOV) 14 cm.The patient was placed in a supine position. Ankle joint localization was maintained using a dedicated coil suited to the shape of the ankle and foot. To further lock the ankle and foot position, elastic wedge-shaped cushions were used.Exclusion criteria were recent fracture (n = 2 from the control group), neoplasm (n = 2 from the control group), sequences without angled axial projections , artifacts obstructing evaluation  and conditions which severely altered the appearance of the ankle . In total, we excluded 44 cases , Fig. Peroneal split tear was defined as a radiologically and clinically proved identifiable longitudinal tear of the PB tendon. All patients that met the inclusion and exclusion criteria were included in the split tear group.not include any PB split tears.The control group was assembled in a similar way. Matching exclusion criteria were applied but, in contrast, the image sequences were evaluated to The soft tissue abnormalities included in the study were abnormality in the SPR, ATFL or CFL. Abnormalities of SPR were classified as total rupture, relaxed or thickened. Abnormalities of the ATFL and CFL were assessed as one of three grades : grade 1The anatomical variations included in the study were the low-lying PB, os peroneum, prominent peroneal tubercle and malleolar groove shape. The presence of os peroneum was defined as a separate bone situated within the PL tendon, typically in the area inferior to the os cuboideum. If the length of the peroneal tubercle, measured from the lateral margin of the calcaneus, exceeded the width of each peroneal tendon it was defined as prominent. Three malleolar groove shapes were included: concave, flat and convex. The PB was considered low-lying if the muscle belly was seen at least at the level of the lateral malleolus at the level where the posterior talofibular ligament (PTFL) attaches. The shape was assessed at the level of where the PTFL attaches to the fibula. Using measurement tools in the Radiological Information System/Picture Archiving and Communication System (RIS/PACS), a straight line could be drawn through the groove. If the groove curved towards the center of the fibula, away from the measurement line, the groove was considered concave. If the groove neither curved nor protruded from the measurement line, it was considered flat. If the groove protruded beyond the measurement line, it was considered convex.To more accurately evaluate localization of bone marrow edema, the tibia, lateral malleolus, talus and calcaneus were divided into sectors see Fig. . The disNo gap between the talus and the adjacent fat pad in the anterior joint recess was defined as no effusion (coded as Effusion 1). Fluid creating a gap between the talus and the adjacent fat pad in the anterior joint recess, as well as fluid expansion of the posterior recess, was defined as moderate effusion (coded as Effusion 2). If fluid also created a gap between the anterior tibia and adjacent fat pad, it was defined as severe effusion (coded as Effusion 3). Presence of the synovitis in the talocrural joint was noted.We measured patients’ height and weight and BMI was calculated. History of trauma was considered if the patient had had an ankle injury in the last 6 months.All MRI examinations were evaluated using the radiology information system/ picture archiving and communication system (RIS/PACS) of the hospital (AGFA©). The data collection procedure was done separately by observers who were radiologists with 12  and 3 years’ experience  and a medical student  who did master’s degrees in medicine at the Department of Musculoskeletal Radiology at SU in Gothenburg, Sweden. PS and AB evaluated the soft tissue lesions. PS, SB and MH graded the bone lesions. Before scoring, the most experienced radiologist (PS) held a training session with all observers to ensure conformity in evaluation. The final decision was made by consensus.The proportion and percentage of feature occurrence, compared to the total number of patients in each group, was calculated . The analyses are based on comparisons of the probability of feature occurrence in both groups. This supposition was made due to the relatively large size of the groups. In this case, the probability was determined by analyzing feature frequency in the study groups. Binominal proportion confidence intervals  were calculated using asymptotic normal approximation and Wilsons’s test, to evaluate variance. The Bonferroni-Holm method (B-H)  was usedContinuous variables (BMI) were found to be normally distributed. Parametric tests were used for variables with normal distribution. Differences in mean values were analyzed with Student’s t-test. A p value ≤ 0.05 was considered as statistically significant.To investigate bivariate correlations, Pearson correlation for binominal variables was used to analyze the direction, strength  and significance of correlations.what differed in the evaluation.To evaluate the inter-rater reliability of the method, the results from the two parties were compared using Cohen’s kappa coefficient for each parameter using SPSS. Cohen’s intervals were used for interpretation: no agreement (≤ 0), none to slight (0.01–0.2), fair (0.21–0.4), moderate (0.41–0.6), substantial (0.61–0.8) and almost perfect (0.81–1.0) . In addiAnalyses and data visualization were performed using Microsoft Excel©, Statistical Package for the Social Sciences program (SPSS)© and Python 3.7.10  in Jupyter notebook.Ethics.The ethical considerations of this study were primarily the management of confidential patient material and the digital storage of said data. Patients were assigned pseudo-encrypted codes through which deidentification was achieved. The Swedish Ethical Review Authority approved the study and waived the need for informed consent (number 2020-06-177 and 2021–05447).The study was conducted in compliance with the Declaration of Helsinki. The anonymization of patient data ensured data protection following the European General Data Protection Regulation. The data were recorded in a password-protected secure database.MRI examinations were assigned into two groups, experimental and control, following the flow chart in Fig. No statistically significant differences were found regarding demographic and anthropometric parameters were similar in both groups (p > 0.05), Tables * As “history of trauma” could not be determined for all patients, in these calculations n = 73 for the split tear group and n = 102 for the control group.The agreement between observers in the experimental group was moderate to perfect ; the higThe most common findings in the experimental group were peroneal tenosynovitis (n = 27), relaxed SPR (n = 29), and thickening of SPR  was moderate to perfect, highest for bone marrow edema in the medial part of the distal tibia  and lowest for the medial part of the calcaneus, Table In the split tear group, the Cohen’s kappa value between the most experienced observers was the lowest for “convex malleolar groove” (moderate) and the highest for concave  Table . In the Agreement in evaluation of os peroneum and prominent peroneal tubercule between the most experienced observers was substantial in the experimental group and moderate in the control group Table .FT- Fisher test, H-B - Holm–Bonferroni method.Most of bone variables are somewhat similar in occurrence in both groups, Fig. Using FT statistically significant differences were identified in occurrence of presence of bone marrow edema in the posterior half of the lateral malleolus, normal and moderate increased amount of joint fluid Table . After uIn Fig. The joint fluid profile illustrates how the control group leans more heavily towards no joint effusion (coded as Effusion 1) and the split tear group towards moderate joint effusion (Effusion 2) Figs.  and 5.The malleolar groove profile demonstrates how the malleolar groove shape was more often concave in the split tear group while the other two shapes were slightly more common in the control group, however no statistical significance was seen Figs. ; Table 3Significant differences in percentual feature occurrence between the two groups can be seen in Table No joint effusion and history of trauma were significantly less common; 95% CI limits ranged from 2% to − 35%  between bone marrow edema in the medial part of the talus and normal joint fluid, a positive correlation (r = 0.32) between bone marrow edema in the anterior part of the lateral malleolus and no joint effusion, and a negative correlation (r = − 0.27) between bone marrow edema in the central part of the distal tibia and severe joint effusion.In the control group, other correlations were predominant, with the strongest being a positive correlation (r = 0.27) between the presence of bone marrow edema in the medial talus and severe joint effusion. A negative correlation (r = − 0.22) was found between bone marrow edema in the medial talus and normal joint fluid. Correlations in the control group were weaker than those in the split tear group; none reached r ≥ 0.3. When the groups were combined, a negative correlation (r = − 0.15) was still found between bone marrow edema in the medial talus and no joint effusion. No correlation was found between bone marrow edema in the anterior fibula and joint effusion. The strongest correlation (0.31) was between medial talus and severe joint effusion. No other significant correlations were found.We found significant differences between groups in the presence of bone marrow edema in the posterior part of the lateral malleolus, while there was no significant difference in occurrence of ATFL or CFL ruptures. Complete rupture of SPR was observed only in the control group. The most significant result of the current study is that bone marrow edema in the posterior half of the lateral malleolus is significantly common in patients with a peroneus split rupture. The absence of other significant differences between the groups confirms that the radiological diagnosis of split rupture is difficult.We found several statistically significant correlations in the split tear group between the presence of bone marrow edema in the medial talus and a normal amount of synovial fluid.Split of the PB is seen more often than that of PL, thus we focus on this pathology , 4. TherBeyond the significant differences, it is worth noting that most bone variables were similar in prevalence. This was generally expected, as most of the features had not been linked to peroneus split tears in previous research. Some features had been though   and findThe magnitude in differences between the groups can be interpreted by observing the CI. They ranged from 2%  to − 35%  where the higher value would carry clinical relevance. Pinpointing the true difference based on our study is thus difficult but it does mean all differences have the potential to be substantial.SPR disorder may cause peroneus tendon pathology inclusive of split rupture. SPR may be weakened or separated from the fibula following inversion ankle injuries. It may result as synovitis, subluxation or luxation of peroneus tendons, and split rupture . InjurieMost split ruptures of the PB are chronic, while ligament injuries are more associated with trauma , 27. TheWe found no differences in the anthropometric variables between the groups. However, it is difficult to determine based on our results whether BMI can contribute to split rupture of the PB. Studies in a more diverse cohort are needed to establish a relationship between BMI and the risk of split rupture of the PB.PB split rupture is multifactorial and can be difficult to diagnose , so accoThe distribution of peroneal groove shapes in neither group matched what was described by Edwards in 1928   which waFrom a pathophysiological perspective, one could speculate why a concave malleolar groove would predispose the peroneus tendons to tear. It is possible that some concave malleolar grooves contain sharp ridges, which in combination with repetitive subluxation or trauma could injure the peroneal tendons. Galli et al.  found coPeroneus split tears most commonly do not cause acute symptoms ; it is lDespite the limited number of subjects, the study still detected statistically significant finding and, relatively speaking, the sample size was substantial. The research to date has tended to only include case series  and hasThere are some limitations to the study. During evaluation of the MR images, we noticed the shape of the malleolar groove changing noticeably in the longitudinal axis. Even when adhering to the defined level of where the PTFL attached, the shape could change from concave in the upper region to convex in the lower region. This would explain why Cohen’s kappa was the lowest for malleolar groove shape. Future studies could use a stricter definition of groove level or, alternatively, use 3D imaging for a more comprehensive view of the malleolar groove. The retrospective character, the lack of surgical correlation and the absence of a sample size calculation may also be limitations of this study.Our study reaffirms that the ankle is one of the most challenging structures to evaluate on MRI. If some of the mentioned adjustments were made, we believe higher inter-rater reliability could be achieved.Split rupture of PB is more common than that of PL , 4. We iThe presence of bone marrow edema in the posterior half of the lateral malleolus should be regarded as a potential “red flag” indicating the presence of PB split rupture. There is seemingly a lack of studies on the area, and we believe this study has laid some groundwork for future research.Bone marrow edema in the posterior fibula is significantly associated with peroneus split rupture. A distinct correlation was found between bone marrow edema in the medial talus and no joint effusion in patients with peroneus split rupture. These could potentially facilitate MRI assessment, enabling faster and more accurate diagnosis for patients, but prospective studies are required for causal corroboration. Beyond a shadow of a doubt, this study suggests that certain MRI features are associated with peroneus tendon split tears.


To solve the current problem of poor weld formation due to groove width variation in swing arc narrow gap welding, an infrared passive visual sensing detection approach was developed in this work to measure groove width under intense welding interferences. This approach, called global pattern recognition, includes self-adaptive positioning of the ROI window, equal division thresholding and in situ dynamic clustering algorithms. Accordingly, the self-adaptive positioning method filters several of the nearest values of the arc’s highest point of the vertical coordinate and groove’s same-side edge position to determine the origin coordinates of the ROI window; the equal division thresholding algorithm then divides and processes the ROI window image to extract the groove edge and forms a raw data distribution of groove width in the data window. The in situ dynamic clustering algorithm dynamically classifies the preprocessed data in situ and finally detects the value of the groove width from the remaining true data. Experimental results show that the equal division thresholding algorithm can effectively reduce the influences of arc light and welding fume on the extraction of the groove edge. The in situ dynamic clustering algorithm can avoid disturbances from simulated welding spatters with diameters less than 2.19 mm, thus realizing the high-precision detection of the actual groove width and demonstrating stronger environmental adaptability of the proposed global pattern recognition approach. Narrow gap gas metal arc welding (NG-GMAW) is a high-efficiency and high-quality welding process for manufacturing thick-section structures ,2,3,4. TIn narrow gap welding, the groove width and weld central position usually vary due to the groove processing error, assembling error and welding thermal deformation, which leads to uneven sidewall penetrations and inconsistent bead surface ,16,17. TTo adaptively control weld formation for the SA-NGW process with variable grooves in the future, an infrared passive visual sensing detection approach and system were developed in the present work to measure groove width under intense welding interferences. This approach detects groove width from disturbed images of welding by global pattern recognition based on the adaptive extraction of the groove edge and the dynamic classification of groove width data. Accordingly, an in situ dynamic clustering algorithm is presented to identify the value of groove width from the classified true data in the data window after accurately extracting the groove edge by an equal division thresholding method. Additionally, simulated welding spatters and actual welding experiments were performed to demonstrate the anti-interference capability and effectiveness of the proposed approach so as to build a foundation for self-adaptive controls of the arc swinging angle and welding deposit in a groove of varying width.wV. Note that the test piece is actually fixed to the worktable during welding.θ. At lower frequencies of several hertz, the torch oscillator periodically turns the bending conductive rod to swing the arc circularly at the wire tip within the narrow gap groove. When the arc swings to every sidewall of the groove, this oscillator synchronously outputs a signal of the arc position once the arc pauses briefly at the sidewall. This arc position signal and the base current signal bi of the pulsed arc simultaneously trigger the camera to collect the image of the welding area. After the image is transferred to the acquisition card and processed by the computer, the groove edge on the opposite side of the arc is extracted to avoid arc light interference, and finally, the value of groove width is detected from the left and right edges of the groove.The positive pole of the power supply is connected to the carbon brush in the torch, and the current sensor detects the current signal of the pulsed arc. The camera is fixed to the torch, the lens of which is axially separated from the wire end by a distance of 270 mm and is directed at the welding pool ahead of the torch at a depression angle of An example of the actual welding infrared image is shown in i indicates the ith frame of the welding image, and Oi1 and Oi−1)2( are the current and previous origins of the region-of-interest (ROI) window. From the current and previous adjacent frames of ROI window images, i.e., the ith and the (i − 1)th frames, the positions RjP  and LjP  of right and left edge points of the groove are extracted, and then the corresponding raw datum j is the ordinal number of the position of h is the height of the square ROI window and expresses an integer value in pixels. Accordingly, the raw data distribution matrix h data, is formed from Ci1  and Ci−1)2 of the ROI window from the estimated positional value Since the positions of arc highest point and groove edge actually vary in welding, the position of the groove-edge ROI window needs to be adjusted in real time to overcome such interferences and to accurately intercept the groove edge image on the opposite side of the arc. Accordingly, in image processing, a global image  will be processed sequentially through Gaussian filtering, global thresholding and a morphological opening algorithm . The curyROI lies merely within ±0.3 mm, the value of the groove width can be approximately obtained directly from the left and right groove edges that are extracted from two neighboring frames of ROI window images.Furthermore, for a groove edge image intercepted by the ROI window, a groove edge is extracted by using a median filter, contrast stretching, the Otsu thresholding algorithm, a morphological closing algorithm and the Canny edge detector  in turn.According to the above self-adaptive positioning of the ROI window, together with the following methods of equal division thresholding (EDT) see  and in sIn SA-NGW, arc light and welding fume uniformly distribute in the welding region because the arc moves in a narrow gap groove, and as a result, the uniformity of image grayscale distribution is greatly affected. Consequently, it is difficult to accurately extract the groove edge using the process of Otsu thresholding segmentation from the whole ROI window image of the groove edge. To improve the extraction accuracy of the groove edge in the window image, an equal division thresholding (EDT) method is proposed. This method vertically divides the ROI window image into several equal partitions and then uses Otsu thresholding to segment each partial image for the purpose of accurately extracting each subsection of the groove edge to form the full edge of the groove.sN of the ROI window image on the extraction accuracy of the groove edge, where dS represents the standard deviation of the positional distribution of groove edge points, and α denotes the slope angle of the fitting line of groove edge points. sN on the standard deviation dS and the slope angle α, where the used ROI window image is cut from the welding image and indicated by the area in the white box in sN of vertical equal partitions increases, the slope angle α increases while the standard deviation dS decreases, and correspondingly, the perpendicularity of the fitting line of the groove edge improves. In particular, when the number sN of partitions is greater than 4, the variations in α and dS become gentle, which indicates that the extracted horizontal positions of groove edge points change little and thus suggests that the extraction accuracy of the groove edge becomes high.sN of partitions is 1, the lower partition of the groove edge contour line extracted after whole-threshold processing develops a pronounced bend, as shown by the white line in α is the smallest, while the standard deviation dS is the largest, which implies that the positional dispersity of groove edge points is the largest. When the number sN of partitions is 4, the whole ROI window image is equally cut into four partitions, whose dividing lines are represented by yellow dashed lines in α increases from 77.55° to 85.79°, while dS decreases from 4.581 to 1.449, and the verticality of the fitting line of the groove edge improves. Compared to the whole-thresholding approach, the division thresholding method with four equal partitions reduces the positional dispersity of groove edge points by ~68.37%, which indicates a significant improvement in the verticality of the groove edge fitting line. Therefore, the vertical equal partition number sN of the ROI window image is taken as 4.For example, due to the influence of arc light and welding fume, when the number The principle of the in situ dynamic clustering (ISDC) algorithm proposed is introduced below through flow charts and mathematical expressions. Dynamic analysis of this algorithm is then presented for intuitive understanding. Subsequently, the adaptivity of this algorithm is investigated by simulated spatters to demonstrate the capability of resisting welding spatter disturbance.To reduce the influence of welding spatter on the accuracies of groove edge extraction and groove width detection, the ISDC algorithm is proposed for detecting groove width. This algorithm performs an in situ global dynamic bipartite clustering of raw groove width data in the data window according to the numerical difference degree and removes false data from the raw data according to outlier criteria, finally utilizing linear fitting or mean calculation to obtain the sampling value of the groove width from the preserved true data in the data window. The algorithm process includes data preprocessing, dynamic clustering and cluster selection, as shown in ud in Equation (3).u is the ordinal number of the position of ud. The frequency duf at which different values of ud appear is then counted to determine its greatest value mk is the ordinal number of the current position of λ denotes the ordinal number of the original position of For the purpose of eliminating the turning-point data in the raw data distribution of groove width and thus improving data separability, a data preprocessing algorithm based on the numerical difference degree between adjacent values is presented. The raw datum m = (h−1) or m ≤ Th, respectively, the raw groove width data are directly used as true data . The two data v_dmax are respectively used as the initial centers of two clusters, as shown in Equation (6).ξ denotes the ordinal number of the original position of A-cluster data distribution matrix B-cluster data distribution matrix n is the iteration number of dynamic clustering.For the preprocessed data distribution matrix λ of the original position of According to the numerical difference degree, which is represented by the absolute difference between each datum in aw and bw are the positional variables of the data in AN and BN are the total numbers of data in The centers of clusters Q of cluster selection. By calculating the numerical difference degree between the two cluster centers .Q is determined by subsequent experiments in By sequencing the data in the matrix As an example, ud (in Equation (3)) of the numerical difference between each datum vd_max (in Equation (5)) of numerical difference degrees between each datum By calculating the degree Q. After eliminating the false data distribution matrix Finally, the corresponding raw datum of groove width at the position quantile of 3/4  is used as the value of Q on the adaptability of the ISDC algorithm was investigated with the occurrence of simulated welding spatters of different sizes, as shown in h of the ROI window. The effect of the cluster selection threshold h, a linear fitting method is used to find the sampling value sG of groove width from sG is derived from the 40th value of h (h = 80) data on the fitting line of groove width; otherwise, this value of sG is approximately calculated by finding the mean value of data in During image processing, the right edge image of the groove is cut by the ROI window from the welding image shown in Q on the sampling value sG of groove width, where Q1/4, Q2/4 and Q3/4 denote the corresponding values of Q at the position quantiles of 1/4, 2/4 and 3/4, respectively, and the method of determining the three values of Q is as described in Q do not influence sG. When this proportion varies within the range of 20% to 50%, the two thresholds Q2/4 and Q3/4 have no effect on sG. When further increasing it to 75%, the threshold Q3/4 only slightly affects sG. In other words, with an increase in the threshold Q, the sensitivity of the ISDC algorithm to welding spatter size decreases, which demonstrates that the ISDC algorithm has the ability to increase resistance to welding spatter interference and thus better adapts to welding environments. Therefore, Q3/4 was chosen as the threshold Q in subsequent experiments.Q3/4 of cluster selection. In this case, each raw data distribution matrix sG of groove width are obtained in the same ways as those in sG of groove width is obtained by the direct linear fitting of raw groove width data and comes from the 40th value of h (h = 80) data on the fitting line of groove width.sG of groove width slightly varies with a maximum fluctuation of only 0.034 mm for the ISDC algorithm, while sG decreases significantly for the DLF method, with a maximum reduction value of 0.659 mm. When increasing the number of spatters from 0 to 3, as indicated in sG is merely 0.024 mm for the ISDC algorithm, while sG decreases markedly in the DLF method, with a maximum decrease of 0.235 mm. Clearly, compared to the DLF method, the ISDC algorithm can stably obtain the sampling value sG of groove width despite great variation in the number and size of welding spatters, thus indicating the good adaptability of the ISDC algorithm to welding environments.When increasing this size proportion from 0 to 75%, as shown in To demonstrate the effectiveness of the GPR approach, several detection experiments of groove width were carried out for a flat-position SA-NGW process under the conditions of constant-width and width-varying grooves. The detection accuracy of the GPR approach was then compared to that of the composite linear fitting (CLF) method, which sequentially combines existing image processing, direct linear fitting and mean filtering algorithms, so as to exhibit the superiority of the proposed approach. Finally, the actual width of the post-welding groove is utilized to validate the suitability of the accuracy evaluation approach.L2 × L3 × L4 (length × width × height), as illustrated in bG and eG are the beginning and ending widths of the groove, respectively, and groove depth L1 is set as half of L4. For the experiment with a groove of constant width, the test piece size was 210 × 50 × 40 mm, and the arc swing angle was 82°, while the values of bG and eG were 14 mm before welding. For the experiment with a groove of varying width, the test piece size was 204 × 70 × 30 mm, and the arc swing angle was accordingly reduced to 64° due to the smaller starting value bG of groove width, while the groove width before welding varied uniformly from 11.8 to 15.9 mm.The experimental conditions are the same as those related to During welding, a global image was captured during the swinging of the arc to remain near each groove sidewall, and finally, 323 frames of global images were continuously collected along the whole weld length. By alternately using the ROI window, 162 and 161 frames of left and right groove edge images, respectively, were intercepted at the opposite side of the stationary arc, where one of the left edge images neighbors its previous or subsequent nearest right edge image. Subsequently, 80 instantaneous sampling positional values of corresponding groove edge points were obtained from each edge image to constitute each Accordingly, 322 sets of sG of groove width is generated from each dG of groove width is subsequently obtained by the amplitude-limiting mean filtering of the last five successive values of sG, as indicated by red hollow circles in sG are calculated by direct linear fitting of 322 sets of dG are obtained after the mean filtering of the last five successive values of sG, as indicated by black hollow squares in dG become very low near the positions of the above spatters, particularly around 14.28 mm and 133.28 mm of the weld length. Compared with the CLF method, the GPR approach significantly reduces the influence of welding spatters on the detected values of groove width. Compared with the planned values of groove width on the blue baseline, the detection error of groove width is within −0.086 to +0.109 mm when using the GPR approach and between −0.465 mm and +0.479 mm when using the CLF method. In addition, the standard deviations of the detected values are 0.035 and 0.115 for the two methods, respectively, because thresholding with four equal divisions in the GPR approach has a higher accuracy of groove edge extraction (see example in When using the CLF method, 322 sampling values of sN = 1 in The related performance comparison is listed in eP that was ~7 mm away from the groove terminal while finishing the collection of welding images. During the transiting period between the igniting point sP and the position of the dashed line in To detect groove width, starting from the position of the dashed line, 270 frames of images in total were alternately intercepted by the ROI window for the left and right edges of the groove; that is, 135 images were collected for each of the left and right edges. Eighty instantaneous sampling positional values of corresponding groove edge points were calculated from each image to form each dG were obtained through the GPR and CLF methods, as indicated by red hollow circles and black hollow squares in Similarly to the above constant-groove experiment, the ISDC algorithm eliminated false data from 269 sets of Compared with the planned values of groove width, as shown by the blue line in Moreover, for a clearer understanding, two groups of welding images, which respectively correspond to groove lengths of around 29 mm and 179 mm in sP and eP indicate the igniting and extinguishing points of the arc in the groove, respectively. As an example, Moreover, with an increase in groove width in the two cases, the average penetrations into the groove sidewall decrease from 0.872 to 0.654 mm; meanwhile, the height of the deposited metal, which is actually the vertical distance between the lowest position of the weld surface and the groove bottom, indicated by the red line in Based on infrared passive visual sensing, the detection approach and system of groove width were developed for swing arc narrow gap welding. This global pattern recognition approach can self-adaptively position the ROI window of groove edges, accurately extract the left and right edges of the groove on the opposite side of the arc by thresholding with four equal divisions and then detect the value of groove width by an in situ dynamic clustering algorithm based on dynamic classification. Experimental results show that the proposed approach has a strong ability to resist actual welding interferences and thus yields high detection accuracies, with detection error ranges of −0.086~+0.109 mm and −0.168~+0.119 mm for constant-width and width-varying grooves, respectively.Based on the system and methods proposed, future work will include the concurrent control of the arc swinging angle and welding deposit so as to adapt to the probable variation in groove width. Accordingly, sidewall penetration will be kept sufficient and consistent by self-adaptive adjustment of the arc swinging angle, while a weld bead of constant height will be obtained by the self-adaptive regulation of welding deposit, finally building an intelligent and automatic system for the SA-NGW process.


Historically, intravenous (IV) antibiotics have been the standard of care for the treatment of bloodstream infections (BSIs). IV antibiotic use predisposes patients to line-related complications and is associated with increased length of stay (LOS) and treatment costs as compared to oral (PO) antibiotics. The purpose of this study was to compare the efficacy and safety of IV therapy versus PO stepdown for uncomplicated Streptococcal BSIs.Streptoccoccus spp. in blood cultures with susceptibilities reported, < 48 hours of positive blood cultures, and were treated for the BSI. Patients were excluded if they had polymicrobial bacteremia, mortality prior to treatment completion, unattainable source control, or a complicated infection. The primary outcome was clinical success, defined as absence of infection recurrence, infection-related readmission, and infection-related mortality at 90 days. Secondary outcomes included individual components of composite endpoint, microbiological success, LOS, IV-line associated complications, and antibiotic-associated adverse events.This single-center, retrospective, non-inferiority study included adults who had Of 79 patients included, 36 patients (45.6%) received IV for the full course of therapy and 43 (54.4%) received PO stepdown. Baseline characteristics were similar between groups. The most common sources were skin and soft tissue (34.6%) and oropharyngeal (13.6%). The median duration of IV therapy in PO stepdown patients was 4.4 days (IQR 2.7 – 6.9). Amoxicillin/clavulanate (44%) and cefdinir (24%) were the most common PO agents. Clinical success occurred in 94% of IV only patients and 91% of PO stepdown patients (p = 0.573). No infection-related deaths or recurrences occurred. Line complications were more frequent in patients receiving IV compared to PO stepdown therapy . LOS was lower in the PO stepdown group .This study strengthens the hypothesis that PO stepdown therapy may be non-inferior to IV therapy for clinical success with a trend towards decreased LOS and line complications. However, this study was not powered to meet statistical significance. We are collaborating with several other centers to determine the true impact of this intervention.All Authors: No reported disclosures.


Fecal microbiota transplantation (FMT) is a promising therapeutic approach for microbiota-associated pathologies, but our understanding of the post-FMT microbiome assembly process and its ecological and clinical determinants is incomplete. Here we perform a comprehensive fecal metagenome analysis of 14 FMT trials, involving five pathologies and >250 individuals, and determine the origins of strains in patients after FMT. Independently of the underlying clinical condition, conspecific coexistence of donor and recipient strains after FMT is uncommon and donor strain engraftment is strongly positively correlated with pre-FMT recipient microbiota dysbiosis. Donor strain engraftment was enhanced through antibiotic pretreatment and bowel lavage and dependent on donor and recipient ɑ-diversity; strains from relatively abundant species were more likely and from predicted oral, oxygen-tolerant, and gram-positive species less likely to engraft. We introduce a general mechanistic framework for post-FMT microbiome assembly in alignment with ecological theory, which can guide development of optimized, more targeted, and personalized FMT therapies.  •Pre-FMT patient preparation has larger impact on strain engraftment than disease•Patient antibiotic pretreatment and lavage increase post-FMT donor strain engraftment•Models indicate leverage for personalized FMT optimization with patient-donor pairing•Engraftment rates not different in responders/non-responders with IBD or melanoma Using strain-level microbiota profiling in patients treated with fecal microbiota transplantation (FMT), Podlesny et al. identify a mechanistic framework for post-FMT microbiome assembly that aligns with ecological theory. Donor strain engraftment is dependent on microbiome and clinical parameters, including antibiotic treatment and bowel lavage, independently of the treated disease. Clostridioides difficile infection (rCDI), with a success rate of ∼90% that surpasses those of conventional antibiotic treatments.,,,Fecal microbiota transplantation (FMT) infuses the microbiota of healthy donor feces into the intestinal tract of a patient in the clinical attempt to treat a microbiota-associated disease.The dynamics of post-FMT microbiota organization and the clinical, host, and ecological factors that govern microbiome assembly in different FMT-treated populations and at the level of individual patients and microbes have not been extensively studied. Previous studies mostly relied on 16S rRNA analysis, which lacks the taxonomic resolution to track specific recipient and donor-derived microbiota members after FMT. This limits our ability to elucidate the ecological principles that govern post-FMT microbiota assembly, as interactions between microbiome members are hypothesized to be most competitive among closely related taxa, including conspecific strains belonging to the same species.,C. difficile infection,Although in many aspects FMT represents an ecosystem restoration approach, ecological framework theories have only recently been applied to understand and predict FMT outcomes.In this study, we characterize post-FMT microbiome assemblies at the strain level in patients who present with a range of disease and treatment backgrounds. We develop generalizable models to predict and control FMT outcomes, determine the relationship between post-FMT microbiota assembly and clinical response, thereby defining the opportunities and limitations of FMT-based precision microbiota therapies and outlining clinical strategies to optimize them.C. difficile infection , the IBDs ulcerative colitis (2 studies) and Crohn disease (one study); metabolic syndrome, type 2 diabetes mellitus, or obesity ; drug-resistant Enterobacteriaceae carriage ; or immune checkpoint inhibitor-refractory melanoma . Post-FMT samples were relatively evenly distributed between patient groups over the first 4 months after FMT (+/−), the FMT application type , as well as the number of FMTs received (which varied between a single and up to 41 applications). All rCDI patients were treated with antibiotics as part of their therapeutic regimens, while each of the ICI, IBD, and MDR groups included at least one study in which antibiotics were used as a bowel cleansing strategy to deplete the resident microbiota and prepare patients for donor microbiota engraftment  B. This aMost strains detected in patients after FMT could be assigned to either the donor or the recipient, or they represented new, i.e., previously undetected, strains or species from environmental or other sources A. The coABx−, IBDABx−, and ICIABx− patients was dominated by recipient strains , followed by recipient-derived strains (12.2% ± 20.1%) A. Howeve strains C. In con for FMT C. The an for FMT D, where ications .Donor strains from the majority of species (62.1% ± 15.4%) did not engraft in patients after FMT . Only 10In summary, donor strain engraftment after FMT appears clinically expandable, including for different medical conditions, using antibiotic microbiota disruption to induce dysbiosis in preparation for FMT, as well as other FMT protocol parameters.In order to gain a conceptual understanding of the factors that determine donor microbiota engraftment after FMT in individual patients, we wanted to determine the effects of both ecological microbiota and clinical FMT parameters on post-FMT patient microbiota assembly. As ecological parameters, recipient and donor microbiota (ɑ/β) diversity, and as clinical parameters, patient pretreatment  and FMT modalities (no. of FMTs) were used as input variables for a generalized linear mixed model (GLMM) to estimate donor-derived strain fractions after FMT per patient  and S5. Of the microbiota variables, donor ɑ-diversity (Shannon index) had the strongest estimated positive effect  on overall donor strain engraftment, whereas recipient ɑ-diversity had an estimated negative effect . Simulations with the GLMM to estimate the specific marginal effects of recipient and donor ɑ-diversity, using alternative, artificial values for the Shannon index, i.e., the highest and lowest value that was detected in any donor from the cohort, indicated that a disproportionately large fraction of strains from high-diversity donors would engraft in low-diversity patients after FMT B. ComposOf the clinical variables, antibiotic pretreatment , bowel lavage , and multiple FMT applications  were all predicted to increase donor strain engraftment based on our GLMM A. Time sIn summary, post-FMT microbiota assembly appears to be driven both by the recipient (ɑ/β-diversity) and donor (ɑ-diversity) microbiota, as well as the FMT procedure, i.e., resident microbiota depletion before FMT (by ABx treatment and lavage) and repeated exposure of the patient to the donor microbiota (multiple FMTs).As the previous model estimated the community-wide (ɑ/β-diversity) determinants of donor strain engraftment for individual patients, we next sought to compare the engraftment probabilities for individual strains from different bacterial taxa in a separate GLMM. This second model outlines the taxonomic and functional boundaries for donor strain engraftment, i.e., whether strains from specific taxa or with taxon-specific properties are more or less likely to engraft after FMT. For this GLMM, we used the same set of variables as described above, but in combination with relative abundance and other species features, such as Gram stain, oral habitat, spore formation, or oxygen tolerance, as estimated using functional databases see . SpeciesBacteroidetes compared with Firmicutes, Proteobacteria, and Actinobacteria (Megamonas (21.7%), Desulfovibrio (16.8%), and Paraprevotella (13.7%) representing the most and Klebsiella, Veillonella, and Haemophilus (<2e-07%) the least likely engrafted genera , spore formation , and oxygen tolerance  were all negatively correlated with engraftment probability  were generally higher for members of the phylum bacteria A. On thed genera B. In gend genera B. Donor-bability . Across In conclusion, the engraftment of specific donor strains appears to be less dependent on microbiome (ɑ/β-diversity) and clinical  parameters, than on associated species properties, such as  lifestyle and (recipient and donor) relative abundance.Firmicutes . Across all simulated recipient/donor pairs, the predicted total number of engrafted donor strains per patient varied in a range of log2(±5)-fold between the worst and best-engrafting donors relative to the actual donor D. DonorsCure rates of ∼90% after FMT have been reported for rCDI in patients after repeated antibiotic treatment failure,The FMT meta-cohort further included a subset of two ICI  and two IBD  studies for which clinical response data were available A. BetweeBetween responders and non-responders from the two analyzed IBD studies, consistent differences regarding donor microbiota engraftment could neither be detected across nor within any of the studies or their subgroups . FurtherFMT shows potential as a therapeutic tool for microbiome-associated pathologies and can be used as an experimental approach to establish causality for alterations of the human gut microbiota that have been associated with disease. However, the relationship between donor microbiota engraftment and FMT outcomes in treated patients has not yet been comprehensively characterized, compared, and correlated with patient parameters, FMT modalities, and clinical outcomes. Moreover, the ecological principles that govern the post-FMT microbiota assembly process are poorly understood. Our analyses identify clinically modifiable factors as targets for FMT outcome optimization and differentiate between two distinct therapeutic goals: (1) to maximize broad, untargeted engraftment of the donor microbiota after FMT with antibiotic patient pretreatment, with likely clinical benefits for the treatment of rCDI; and (2) to increase the engraftment probability of specific donor strains by increasing the relative abundance of the corresponding species in the transplant sample or matching a recipient to an optimal donor. The second goal offers the prospect of developing personalized FMT applications for the treatment of different microbiota-associated pathologies in a framework of precision medicine.,A key finding of our study is that donor strain engraftment after FMT is strongly dependent on recipient microbiota composition and dysbiosis. In rCDI, where dysbiosis is rampant  and linkEcosystems assemble through a combination of deterministic, neutral, and historical processes.in situ evolution or genetic adaptation to the host environment) mechanisms,,Findings from our strain-level analysis provide insights into the ecological principles that shape the post-FMT patient microbiome, as well as the influence of clinical factors on these principles, specifically as they relate to (1) the conspecific coexistence of strains, and (2) the ratio by which donor strains engraft at the expense of recipient strains. In a community that is shaped by deterministic, niche-specific processes, such as the competition between members based on fitness differences, coexistence theory would postulate that closely related taxa with resource niche overlaps, such as conspecific strains, are more likely to compete than to coexist.Although we cannot rule out the possibility that a reduced sensitivity of our bioinformatic analysis toward low-abundant strains is responsible for the decreased detection of strain coexistence, our findings are in agreement with strong niche-related competition and priority effects favoring recipient strains in undisturbed communities. Depletion of the resident microbiota is necessary to remove recipient strains, free ecological niches, and effectively overcome priority effects, resulting in increased donor strain engraftment in rCDI patients or recipients pretreated with antibiotics and bowel cleansing. The ability to overcome priority effects through antibiotic treatment has recently been demonstrated for consecutive strain colonization experiments in gnotobiotic mice.Previous microbiome studies mostly lacked the taxonomic resolution to assess the role of donor strain engraftment for the clinical response to FMT in patients with microbiota-associated immunological or metabolic diseases, such as IBD and ICI. We compared donor strain engraftment in responders and non-responders from four IBD and ICI studies and did not observe differences in donor strain engraftment in ICI and IBD responders and non-responders. Moreover, although donor engraftment rates substantially varied between studies , clinicaBlautia producta strains that inhibit vancomycin-resistant enterococci.Studies are beginning to outline personalized FMT strategies by drawing attention to both recipient and donor microbiome features to predict and optimize clinical FMT outcomes.In summary, our findings characterize the contribution of donor-derived strains to the post-FMT patient microbiota across a diverse set of patient, microbiome, and clinical conditions. They suggest a major impact of adjustable patient pretreatment modalities on donor strain engraftment and present generalizable predictive models for patient and strain-specific donor microbiota engraftment that are in agreement with ecological theory. They further illustrate the theoretical potential for personalized FMT applications through fecal supplementation with select strains and recipient/donor matching to increase the engraftment probability of specific strains. With this work, we lay the groundwork for future developments of precision microbiota modulation therapies.,Most clinical trials that were included in the analysis involved small patient numbers, which were sampled at different and inconsistent time points. Similarly, the control dataset of fecal metagenomes from healthy individuals was significantly larger than the patient or donor datasets from the FMT meta-cohort. Although our models identified strong and distinctive associations, these were partially based on study and patient subsets from the meta-cohort, and the temporal trajectories of the post-FMT assembly process were not characterized in detail. However, the use of a heterogeneous, but extensive, meta-cohort was also instrumental in the development of robust predictive generalized linear mixed models and the identification of what appear to be the universal drivers of post-FMT microbiota assembly. Our predictions for donor strain engraftment in other recipient/donor combinations or with fecal samples supplemented with bacterial cultures should be experimentally tested in follow-up clinical trials, as our retrospective study design did not allow for independent validation of our simulated FMT outcomes. Contributions of non-bacterial microbes, such as fungi and viruses/phages, to the post-FMT microbiome assembly, which may be clinically relevant,w.florian.fricke@uni-hohenheim.de).Further information and requests for resources and reagents should be directed to and will be fulfilled by the lead contact, W. Florian Fricke  and their donors and to apply strain-level microbiota profiling in order to determine donor-derived strain contributions to post-FMT microbiota assembly. This information was used to inform predictive models to describe the post-FMT microbiota assembly process and to determine the role of recipient and donor microbiota features, taxonomic and ecological microbiota parameters, and clinical modalities of patient preparation and FMT for donor strain engraftment.Clostridioides difficile infection (rCDI),,,,,,,Enterobacteriaceae carriage (MDR),,,We assembled a comprehensive meta-cohort of available metagenomics data from clinical FMT trials, including 254 distinct clinical cases in which FMT was used to modulate the microbiota of patients with different medical conditions A. PublisClinical response information was obtained from the original publications and defined as follows: For ICI patients . To remove host contamination, trimmed reads were mapped against the human genome (GRCh37/hg19) with Bowtie2 v2.2.3.,Taxonomic analyses were carried out to provide an overview of sample microbiota compositions and to generate the marker gene alignments that served as input for the strain-level analysis with SameStr. Preprocessed sequence reads from each sample were mapped against the MetaPhlAn clade-specific marker gene database  was incorporated as a fixed effect, using samples from the control cohort as a reference. Only post-FMT patient sample data were included that were collected at least five days after FMT. We controlled for study effects in the control cohort and repeated post-FMT patient sampling by including study and case as random effects. Multicollinearity was assessed by calculating the variance inflation factor with the performance package v0.7.2 , individual donor samples that the authors had combined in batches for FMT were combined. For Li et al. (BOR), which included data from a single donor sampled at three distinct time points without providing information about their specific use for distinct patients, donor samples were combined, as described in the original publication. For Kump et al. (GOR), which included repeated FMT, all available sequence data for each donor were combined across treatment rounds. Ng et al. (CHA) did not disclose sample metadata in the publication, including information about patient/donor pairing and patient assignments to different treatment groups , and this information was not made available upon requests to the authors. For this study, concatenated sequence files of all five donor samples were used and sham cases identified and excluded based on the lack of shared strains with any post-FMT sample see . In caseGeneralized linear mixed-effects models (GLMMs) were used to estimate the effects of recipient and donor microbiota parameters and clinical modalities on the post-FMT microbiota assembly process. Separate GLMMs were used to determine the role of these parameters for donor-derived strain fractions in individual patients after FMT  and of tOverall donor microbiota engraftment  was calc,We expanded our model to estimate the relevance of relative abundances and functional features  on the engraftment probabilities of individual strains in the meta-cohort . Using all species that were detected in a donor and provided only with donor and pre-FMT patient microbiota information, the model predicted whether the corresponding donor-derived strain would be detected in the post-FMT patient or not. Functional features were coded on the species level as +1  and −1 , replacing missing values with each feature’s mean frequency across all species (in order to avoid an influence on the model). Since some donors were used to treat multiple patients, in addition to repeated sampling and study effects, we also controlled for donor-specific influences with random effects in our model. The adjusted engraftment probability of each genus  is shownThe GLMM for the engraftment of individual strains  closely fit the underlying data, both for predicted donor strain engraftment events of individual genera and aggregating total numbers of engrafted strains . Therefore, we used the model to simulate FMT outcomes for other recipient/donor pairs from the meta-cohort. For these simulations, species-level taxonomic compositional profiles of recipient and donor samples, as determined with MetaPhlAn3, were used as input for the GLMM and the predicted total number of engrafted donor strains  were aggregated for each patient and time point and compared to actually detected numbers from the real recipient/donor pairs by calculating (log2) fold-changes D.https://www.github.com/danielpodlesny/SameStr).The code for SameStr, including documentation, examples and notebooks, is available at Github (


TT) derivatives bearing one and three hexyl-thiophene moieties, TT-HThio and 3TT-(HThio), have been prepared and characterized. Both compounds display enhanced quantum yields in their crystalline form with respect to those in a solution state, revealing crystallization-enhanced emissive (CEE) behavior. Importantly, while single fluorescence is observed in solution, crystalline powders also feature dual ORTP, whose respective molecular and aggregate origins have been disclosed through X-ray diffraction analysis and DFT/TDDFT calculations. The relation between the photophysical properties of TT-HThio and its crystallinity degree has been confirmed by a decrease in photoluminescent quantum yield (Φ) and loss of vibronic resolution when its crystals are ground in a mortar, revealing mechanochromic behavior and confirming CEE features.The development of organic room-temperature phosphorescent (ORTP) materials represents an active field of research due to their significant advantages with respect to their organometallic counterparts. Two cyclic triimidazole ( Single-component organic materials characterized by rich emissive behavior, including room-temperature long-lived features, are receiving growing attention from the scientific community due to the benefits they offer with respect to their widely used metal-containing phosphorescent counterparts, including biocompatibility and low cost. Applications of organic room-temperature phosphorescence (ORTP) in different fields spanning bioimaging ,2, anti-a:1’,2’-c:1’’,2’’-e]triazine, TT triazine, TT-HThio, and 3,7,11-tri(4-hexylthiophen-2-yl)-triimidazotriazine, 3TT-(HThio) -triimidazotriazine, TT-HThio, and 3,7,11-tri(4-hexylthiophen-2-yl)-triimidazotriazine, 3TT-(HThio), have been synthetized by Stille coupling between tributyl(4-hexylthiophen-2-yl)stannane and the corresponding mono- (TTBr) or tri-brominated (3TTBr) TT derivative (3-(4-hexylthiophen-2-yl)-triimidazo[1,2-rivative .−5 M), TT-HThio displays a single fluorescence (FL) with photoluminescent quantum yield, Φ, equal to 11%; wavelength of the maximum peak (λem) = 370 nm and lifetime (τ) = 0.81 ns at 298 K; and λem = 365 nm and τ =1.71 ns at 77 K (TT-HThio), one fluorescence  and 77 K, an excitation-dependent PL behavior comprising one fluorescence and two phosphorescences , one triplet excited state of molecular origin , Tmol, and one triplet excited state of aggregate origin , Tag, are observed. A mirroring relationship with the corresponding emission is clearly visible at 77 K.Crystals of escences . In partTT-HThio crystals, we hypothesized possible aggregation-enhanced emissive (AEE) features; therefore, solvent (THF)/non-solvent (water) experiments have been performed. Addition of increasing water fractions to TT-HThio in THF  results, instead, in emission quenching  and the loss of vibronic components when features .−5 M), 3TT-(HThio) displays a single fluorescence ), one fluorescence  has been performed on crystalline powders (Φ = 22%), while different attempts to prepare single crystals suitable for XRD analysis failed. Similarly to TT-HThio, 3TT-(HThio) displays at both 298 and 77 K an excitation-dependent PL behavior, comprising FL, HEP and LEP , one Tmol , and one Tag  are visible both at 298 and 77 K. The photophysical parameters of TT-HThio and 3TT-(HThio) are summarized in The solid-state photophysical characterization of  and LEP . In partTT-HThio suitable for X-ray diffraction analysis have been obtained by the slow evaporation of methanol solutions. TT-HThio crystallizes in the P-1 space group with two molecules in the asymmetric unit, A and B, forming very long colorless needles. In both A and B molecules, the TT and Thio moieties are almost coplanar, the least-squares planes through the two units are 14.12 and 6.00° in A and B, respectively , S1A⋅⋅⋅C9Ax,y,z1+, 3.344(2) and C5A⋅⋅⋅C5Ax,1−y,1−z1−, 3.376(3) Å (A stack) and S1B⋅⋅⋅C9Bx,y,z1+, 3.388(2) Å (B stack). The TT moiety of molecule A is connected from both sides to other two centrosymmetry-related A molecules through quite short C–H⋅⋅⋅N hydrogen bonds , forming infinite ribbons from which the thio-hexyl chains depart  connects A and B molecules with each other. Single crystals of ectively a. The heectively b, both fs depart c. The coTT scaffold, the presence of the chromophore switches on fluorescence which is otherwise absent in TT itself in solution. From DFT/TDDFT calculations, it was inferred that TT lacks low-energy singlet states with non-null oscillator strength (f) due to the high symmetry of its electronic system. Analogously, “silent” low-energy singlet states are present in trihalogenated TT derivatives [TT derivatives, the disruption of the molecular symmetry results in S1 and upper levels with medium-to-high oscillator strength depending on the nature of the substituent itself. This is also noted for TT derivatives functionalized with three equal chromophoric fragments owing to the adopted non-planar conformation [TT-HThio, the S1 level is computed at 258 nm (f = 0.49), quite similar to the mono-pyridine TT derivative for which S1 is computed at 256 nm with f = 0.48. In both cases, S1 is mainly (88%) a HOMO→LUMO transition of  type and very weak CT character (in the present case from thiophene to TT)  are computed at 258 , 257  and 257 nm , all of them of  type with very weak CT character.As previously observed for a series of TT derivatives bearing one to three chromophoric substituents  . Moving o TT)  can be explained by examining the singlet–triplet energy gap, ΔEST, separating S1 and the closer, lower-energy triplet state (Tn) and the nature of this level. In the DFT freely optimized geometry of TT-HThio, starting from its X-ray molecular structure, a rather high (0.53 eV) energy gap is obtained. However, compared with S1, the closer triplet state, T5  transition) has a much more marked CT character, as deduced by looking at the involved HOMO-2 (1) and 3.37 D (T5). The different character of S1 and T5 suggests an easy intersystem crossing (ISC) which explains the HEP observed in crystals. Moreover, it should be noted that the optimized structure is characterized by much larger twisting than that observed in the X-ray structure. In fact, the C1-C2-C10-S1 torsion angle  and 0.27 eV (ω = 7.4°), making the singlet-to-triplet ISC even easier. HEP is therefore associated with radiative decay from T5 itself or a lower triplet state Tmol after internal conversion (IC) from T5. It is visible in crystals thanks to the rigidifying effect and protection from oxygen quenching ascribable to intermolecular interactions. The observation of HEP in the solid state for both d HOMO-2 , which iTT-HThio and 3TT-(HThio) is reported in the Jablonski diagram below , they are responsible for a different emissive behavior in their solid state. While crystals of both compounds display a high-energy phosphorescence of molecular origin , associated with emission from a 3 low-energy triplet level, TT-Py shows an additional long-lived molecular component, almost overlapped with the fluorescence. This latter emission was ascribed to radiative decay from a triplet state of mixed 3 symmetry, which is responsible for the observed dual anti-Kasha phosphorescence [TT-HThio, whose frontier MOs have all π symmetry. Moreover, crystals of TT-Py display an additional low-energy fluorescence, associated with the higher mobility of the chromophoric pendant compared with the hexylthiophene group. The additional phosphorescent emission, LEP, can be associated with the presence of H-aggregates in the crystal structure, in agreement with previous findings on compounds with the same triazinic scaffold ,25,26,27am below . It is irescence . Such trTT family have been isolated and characterized. The compounds, bearing one or three hexylthiophene fragments, display CEE behavior comprising fluorescence and heavy-atom-free dual phosphorescence, associated with molecular and supramolecular features. This work adds a new building block to the knowledge of this family and to RTP organic phenomena in general. From these results, the preparation of new multicomponent emitters can be envisaged. In summary, two new members of the photophysically intriguing TTBr and 3TTBr were prepared according to published procedures [1H and 13C NMR spectra were recorded on a Bruker AVANCE-400 instrument (400 MHz). Chemical shifts are reported in parts per million (ppm) and are referenced to the residual solvent peak ; coupling constants (J) are given in hertz (Hz) and are quoted to the nearest 0.5 Hz. Peak multiplicities are described in the following way: s, singlet; d, doublet; t, triplet; p, pentet; m, multiplet. All reagents were purchased from chemical suppliers and used without further purification unless otherwise stated. ocedures ,21. Tribocedures . 1H and TT-HThio and 3TT-(HThio) dispersed in polymethylmethacrylate (PMMA) were prepared by spin coating  a dichloromethane solution (TT-HThio or 3TT-(HThio)/PMMA = 0.5 wt %; PMMA = 10 wt % with respect to the solvent) on a quartz substrate.Films of TT-HThio was prepared by Stille coupling between TTBr and tributyl(4-hexylthiophen-2-yl)stannane. In a typical reaction, TTBr , tributyl(4-hexylthiophen-2-yl)stannane , LiCl , Pd(PPh3)2Cl2  and dry toluene (10 mL) are transferred inside a 100 mL dry Schlenk flask equipped with a magnetic stirrer. The mixture is de-aerated by means of three freeze–pump–thaw cycles. The system is heated under a static nitrogen atmosphere at 120 °C for 16 h. The reaction is then cooled to room temperature, filtered on Buchner and the solvent removed in vacuum. The crude reaction mixture is purified by flash chromatography on SiO2 with DCM/ACN as eluents (Rf = 0.53 in DCM/ACN = 8/2). The product is further purified by precipitation from DCM with hexane to give TT-HThio as a white solid . Crystals suitable for X-ray diffraction analysis were obtained by slow evaporation from a MeOH solution.2Cl2, 298 K, δ, ppm): 1H NMRNMR data , 7.53 , 7.26 , 7.22 , 7.07 , 2.69 , 1.73 , 1.46–1.30 , 0.95 . 13C NMR 144.3 (C), 136.4 (C), 136.0 (C), 135.8 (C), 132.3 (CH), 129.6 (CH), 129.1 (CH), 129.1 (CH), 128.1 (C), 123.1 (C), 122.3 (CH), 112.1 (CH), 111.7 (CH), 32.3 (CH2), 31.0 (CH2), 29.6 (CH2), 23.2 (CH2), 14.4 (CH3).3TT-(HThio) was prepared by Stille coupling between 3TTBr and tributyl(4-hexylthiophen-2-yl)stannane. In a typical reaction, 3TTBr , (tributyl(4-hexylthiophen-2-yl)stannane , LiCl , Pd(PPh3)2Cl2  and dry toluene (10 mL) are transferred inside a 100 mL dry Schlenk flask equipped with a magnetic stirrer. The mixture is de-aerated by means of three freeze–pump–thaw cycles. The system is heated under a static nitrogen atmosphere at 120 °C for 16 h. The reaction is then cooled to room temperature, filtered on Buchner and the solvent removed in vacuum. The crude reaction mixture is purified by flash chromatography on SiO2 with DCM/hexane as eluents (Rf = 0.72 in hexane/AcOEt = 8/2). The product is further purified by precipitation from DCM with MeOH to give 3TT-(HThio) as a white solid . Crystals suitable for X-ray diffraction analysis were obtained by slow evaporation from a MeOH solution.2Cl2, 298 K, δ, ppm): 1H NMR NMR data : 1H NMRNMR data , 7.18 , 7.09 , 2.68 , 1.68 , 1.48–1.30 , 0.90 . 13C NMR 143.9 (C), 136.4 (C), 132.3 (CH), 129.2 (CH), 128.0 (C),122.3 (CH), 122.3 (C), 32.1 (CH2), 30.8 (CH2), 29.40 (CH2), 23.0 (CH2), 14.3 (CH3). TT-HThio were collected on a Bruker Apex II diffractometer  using MoKα radiation [2 using all data. Hydrogen atoms were placed at geometrically estimated positions. Details relating to the crystal and the structural refinement are presented in TT-HThio were grown at room temperature in a methanol solution of the compound. Full details of crystal data and structure refinement, in CIF format, are available as X-ray data of adiation . The stradiation  using a TT-HThio and 3TT-(HThio) were performed with the Gaussian 16 program (Revision A.03) [TT-HThio was optimized starting from the experimental molecular structure, as derived from X-ray studies. For comparison purposes, we adopted the same functional ωB97X [DFT and TDDFT calculations on on A.03)  using thal ωB97X  used forav = 2 quartz dewar.Photoluminescence quantum yields were measured using a C11347 Quantaurus–Absolute Photoluminescence Quantum Yield Spectrometer  equipped with a 150 W Xenon lamp, an integrating sphere and a multichannel detector. Steady-state emission, excitation spectra and photoluminescence lifetimes were obtained using an FLS 980  spectrofluorimeter. The steady-state measurements were recorded with a 450 W Xenon arc lamp. Photoluminescence lifetime measurements were performed using an EPLED-300 (Edinburgh Instrument Ltd.) and microsecond flash Xe-lamp  with the data acquisition devices time-correlated single-photon counting (TCSPC) and multi-channel scaling (MCS) methods, respectively. Average lifetimes were obtained as τ


The TEVA were more frequent in bilateral UDT (p = 0.009). Multiple defects were observed more frequently in the intra-abdominal testicles (p = 0.028). A correlation was found between the testicular atrophy index (TAI) and the incidence and number of TEVA in the UDT boys (p < 0.001). The smaller the testis (higher TAI), the more the defects that appeared in it and the higher the frequency of their appearance. Another correlation was established between testis position and the incidence and number of TEVA (p < 0.001). The higher the testis position, the more the defects that appeared in it and the higher the frequency of their appearance. A correlation was established between the position and the volume of the affected testis (p < 0.001). The higher the gonad position, the more severe the atrophy observed in it. The TEVA were more frequent in the UDT boys than in the hydrocele patients. We revealed that the risk of abnormal fusion between the testis, epididymis, and vas deferens is connected with the testis position  and bilateral non-descent.The goal of this study was to determine the prevalence of the testicular, epididymal, and vasal anomalies (TEVA) in cryptorchid and communicating hydrocele pediatric patients. Six hundred and ninety-one prepubertal boys underwent inguinal exploration for 741 undescended (UDT) or hydrocele testes. Two hundred and fifty-five TEVA were detected in 154 UDT boys, compared to 32 defects in 24 hydrocele patients ( In human beings, as in most mammals, the testis descends from the abdomen to the scrotum  to find a lower ambient temperature for normal spermatogenesis . The tesThe pathogenesis and etiology of cryptorchidism, although it has been extensively studied, is still unclear. Among multiple casual factors related to the condition, anatomical variations, including patent processus vaginalis (PV) and testicular/epididymal fusion anomalies, have been reported in cryptorchid patients, with a range from 32 to 72% ,5. HowevCryptorchidism is frequently 66.7–73%) associated with patent PV 6.7–73% a,9.Testicular and epididymal appendices have been considered to be congenital anomalies . They arThe goal of this study was to determine the prevalence of the testicular, epididymal and vasal anomalies (TEVA) in cryptorchid patients compared with boys with a communicating hydrocele and to examine the correlation between the presence of TEVA and the position of the undescended testis.We included 691 boys (aged 1.5 month–9 years) who were undergoing inguinal exploration for cryptorchidism and a communicating hydrocele in the Department of Pediatric Surgery and Urology at the University Center of Pediatrics in Lodz between January 2011 and December 2017. No patient was excluded from the study.All the patients underwent clinical and ultrasound examination before the treatment. The position and the three dimensions of the testis were recorded and used to calculate the testicular volume (TV) and testicular atrophy index (TAI) of the affected testis . The TAIThe patients were divided into age subgroups based on the developmental changes in testes: 0–2 years, 3–6 years, and 7–9 years  Table 1Table 1. A total of 741 testes were operated on. During the surgery, via an inguinal approach, the spermatic cord was isolated, the hernial sac or PV was dissected, when present, and high ligation was performed. In cryptorchid patients, the testis’ location and size were examined; orchidofunicolysis and orchidopexy were performed by the conventional technique of placing the testis in the subdartos pouch [Patients with hydrocele were qualified for operation after twelve months of age. Early surgery was performed in boys with a concomitant inguinal hernia or a large persistent hydrocele hindering the daily care of a child .The testis, epididymis, and vas deferens were meticulously examined, and any detected anomalies were recorded and classified as major (occluded sperm pathways—defects that could impair patient’s future fertility) or minor (patent sperm pathways—not influencing future fertility) .The anatomy of the gonads was regarded as normal when a normal firm attachment between the testis and the caput and cauda epididymis, as well as a connection to the vas deferens, was present. The anomalies were classified as a separation of the caput and/or cauda epididymis from the testis a,b or a t-test and Pearson’s chi-squared test were used for comparison of the examined parameter values and the incidence in the studied groups of patients. Differences were considered significant at p < 0.05.All the analyses were performed using Statistica 13.1 for Windows . The distribution of the data was analyzed using the Shapiro–Wilk test. The data were distributed in a nonparametric manner; so, they were presented as median and range and analyzed using the ANOVA Kruskall–Wallis test for the assessment of the statistical difference between the groups. The Spearman r correlation was conducted between the results of the examined parameters. The Student A total of 255 anomalies of the testis, epididymis, and vas deferens were detected in 164 UDT boys, compared to 32 defects in 24 patients with hydrocele .p < 0.001). Conversely, the incidence of appendices testis/epididymis was significantly higher in hydrocele patients   was significantly higher in both UDT groups in comparison to the hydrocele group  .p < 0.001). By analogy, the incidence of the minor defects also decreased with the lower position of the testis   . Conversp < 0.001) and the hydrocele group , while in the intra-abdominal UDT group testicles with multiple defects were observed more frequently .In 133 UDT patients, there was 1 defect only; in 17 patients, there were 2 defects; and in 14 patients, 3 defects were detected. One anomaly was observed in 16 boys, and 2 were observed in 8 boys with hydrocele . Testes p < 0.001) and canalicular  groups, as well as in the overall UDT group   .. The smp < 0.001). The higher the gonad position, the more the defects that appeared in it and the higher the frequency of their appearance.A significant correlation was also established also between the position of the testicle and the incidence and the number of TEVA .In the intra-abdominal UDT group, all unilateral and bilateral gonads were affected . Howeverp < 0.001) (p < 0.001). The higher the gonad position, the more severe the atrophy that was observed in it.The TAI of the undescended testes was significantly higher for the intra-abdominal gonads (< 0.001) . A signiDirect comparisons of the healthy and undescended testes’ size between the groups were not performed as these would have been directly dependent upon the patients’ age. The TAI seems to be the best tool for comparison, indicating the percentage loss of the affected testicle volume in relation to the healthy testicle, regardless of the absolute volume of the testicle related to the patient’s age.The sexual differentiation of male internal sex organs occurs in a narrow window of time between 8 and 12 weeks of human fetal development . Leydig Our results are consistent with those of other authors who have shown that cryptorchidism can be associated with various anatomical anomalies, but epididymal anomalies and patency of the PV are among the most frequent ,5,21. RaMoreover, we revealed that the higher the position of the testis the greater the incidence of TEVA. The same observation was made by Caterino et al. , who fouThus, we may assume that some factors secreted by developing testes (predominantly testosterone) are required not only for the epididymal development, but also for proper connections with the testicular efferent ducts, as well as PV obliteration. The expression of androgen and estrogen receptors in the epithelia of the early efferent ducts has been well documented . HoweverIn our study, we observed that boys with hydrocele show, although less frequently, TEVA as an accompanying disorder. This was observed by other authors as well ,5. In tuIn our study, we confirmed that TEVA are frequent in boys with UDT, more than in boys with hydrocele. We revealed that the risk of abnormal fusion between the testis, epididymis, and vas deferens correlates with the position and hypotrophy and with the bilateral non-descent of testis and is the highest in intra-abdominal, hypotrophic, and bilateral undescended testes. These abnormalities may be related to underlying testicular dysgenesis and androgen deficiency in utero rather than to the patent PV. However, this observation is not the explanation for the pathogenesis of UDT and TEVA in all cases. As major TEVA may impair future fertility, patients with a history of cryptorchidism should be under the care of an andrologist after the age of 18.


Brucellosis is among the most widespread zoonotic diseases worldwide. Although rare, nervous system involvement due to Brucella infection is a major diagnostic challenge in endemic regions.This study was a cross-sectional investigation of hospitalized adults with neurobrucellosis from March 2007 to February 2017. We described the clinical characteristics, radiographical and laboratory features, and clinical outcomes of patients with neurobrucellosis.Fifty-four patients with neurobrucellosis were included. The median age was 35  years, and 32 (59%) cases were male. Thirty-four (63%) patients were stockmen or shepherds. The most common clinical manifestations were fever in 49 (91%) cases, headache in 47 (87%), decreased consciousness in 12 (22%), and seizures in 6 (11%). Meningeal signs were detected in 36 (67%) cases. Brucella species were isolated in five cases from blood or cerebrospinal fluid (CSF). The median of CSF leukocytes was 75 per µL, CSF protein 83 mg/dL, and CSF glucose 39 mg/dL. Only two cases had severe hypoglycorrhachia and one CSF protein ≥ 500 mg/dL. No patient died during hospitalization.The symptoms of neurobrucellosis could be mild and nonspecific and the classic triad of meningitis is uncommon. Mild CSF pleocytosis of fewer than 50 leukocytes per microliter of CSF was common but severe hyperproteinorrhachia and severe hypoglycorrhachia were rare in neurobrucellosis. Differentiation between neurobrucellosis and systemic brucellosis is important, because more prolonged treatment is indicated for neurobrucellosis, and it could be associated with a broad spectrum of complications that require close follow-up. Although the incidence of human brucellosis has been reduced to a low level in western European countries and the United States and eradicated in some countries , 2, it iNervous system involvement is an infrequent complication of brucellosis that occurs in 3–10% of patients with brucellosis –8. AlthoDiagnosis of neurobrucellosis among laboratory-confirmed cases of brucellosis is confirmed by isolation of brucella from CSF and/or positive anti-Brucella antibodies in CSF . HoweverDespite a low mortality rate, neurological sequelae remain frequent after neurobrucellosis . It has The study was a cross-sectional investigation that was conducted in two main referral centers for CNS infections in Mashhad, Iran. Mashhad is the second largest city in Iran that is located in the northeast of the country. All individuals (≥ 15 years) who were diagnosed with neurobrucellosis between March 2007 to February 2017 were included in the study. Information regarding age, sex, job, history of consumption of unpasteurized dairy products, clinical characteristics, radiographic and laboratory features, and clinical outcome were recorded retrospectively using a checklist.Brucellosis was diagnosed based on suspected clinical findings and isolation of Brucella species from blood culture and/or positive serum anti-brucella antibodies.Neurobrucellosis among laboratory-confirmed cases of brucellosis is diagnosed by the presence of any one of the following criteria: (1) suspected symptoms and signs of neurobrucellosis such as severe and persistent headache that interferes with the patients normal activity, insomnia, confusion, depression, behaviour change, incontinence, and neck stiffness, and any neurological finding in examination (2) isolation of brucella from CSF and/or positive anti-Brucalla antibodies in CSF, (3) presence of lymphocytic pleocytosis, elevated protein and decreased glucose levels in CSF, or (4) cranial MRI or CT scan findings .Over the period of the study, conventional culture media were used for CSF culture and standard BACTEC™ Plus Aerobic/F bottles for blood cultures. Conventional culture media were incubated for 14–21 days and BACTEC bottles for 1 week., and 1/40 in 2ME (2-Mercaptoethanol) test were considered positive, considering other important differential diagnosis and ruling them out. In CSF, a cutoff titre of 1/8 in Wright test was considered positive. Patients with neurobrucellosis were treated with a triple-drug regimen including ceftriaxone, doxycycline, and rifampin for 4 weeks, followed by doxycycline and rifampin for at least 4 months. In those who did not receive or complete a 4-week parenteral ceftriaxone, trimethoprim–sulfamethoxazole was included in the initial regimen as the third agent.According to the Iranian national protocol for diagnosis and management of brucellosis in the endemic areas, the cutoff titre of 1/80 in Wright, 1/40 in Coombs testsContinuous data were described with median and interquartile ranges and categorical variables with frequency and percentage.The ethics committee of Mashhad University of Medical Sciences approved this study with the code of IR.MUMS.MEDICAL.REC.1397.138.From March 2007 to February 2017, 54 cases of CNS infections caused by Brucella species were diagnosed. The median age of patients was 35  years, with an age range of 1–69 years, and 32 (59%) cases were male. Thirty-four (63%) patients were stockmen or shepherds and 51 (94%) mentioned the consumption of unpasteurized dairy products , followed by headache in 47 (87%), nausea and vomiting in 35 (65%), decreased consciousness in 12 (22%), seizures in 6 (11%), and behaviour changes in 8 (5%). Meningeal signs were positive in 36 (67%) cases, focal neurologic deficits in 7 (13%), and papilledema in 6 (11%). Neurologic deficits included cranial nerve (CN) III palsy, bilateral CN VI palsy, combined CN III and VI palsy, and sensorineural hearing loss (CN VIII), each in one. One patient had left hemiparesis, two had decreased visual acuity, and one experienced total blindness. A classic triad of meningitis was found in seven (13%) cases.Brucella species were isolated in four cases, including two cases with positive blood cultures, one with positive CSF culture, and one with both blood and CSF cultures Table . The orgThe median of CSF leukocytes was 75  per µL and 24 (44%) cases had less than 50 CSF leukocytes per µL. Nine (17%) cases had CSF leukocytes ≤ 5/µL. The predominance of polymorphonuclear cells was found in 6 (11%) cases. The median of CSF protein and glucose were 83  mg/dL and 39  mg/dL, respectively. Only two cases had severe hypoglycorrhachia, defined as CSF glucose ≤ 10 mg/dL and one, CSF protein ≥ 500 mg/dL.Neuroimaging studies including brain CT and/or MRI were performed in 47 cases of whom 6 13%) showed abnormalities. Abnormal neuroimaging findings included meningeal enhancement on post-contrast T1-weighted MR images  in a previous report of 48 cases of neurobrucellosis in Turkey , as compAbout half of our patients with neurobrucellosis had mild CSF pleocytosis of fewer than 50 leukocytes per microliter of CSF, including 17% with five or fewer leukocytes per microliter. Previous studies showed that in cases with CSF leukocytes fewer than 50 per microliter, non-infectious neurological disorders should be considered as the main differential diagnoses ; howeverB. melitensis [The yield of Brucella culture from CSF of patients with neurobrucellosis is low (5–30%)  and mostlitensis .Neuroimaging findings in neurobrucellosis are variable and can resemble other infectious or inflammatory conditions. Four patterns of CNS involvement on imaging have been described in patients with neurobrucellosis: normal, inflammation, white matter changes and vascular changes. Inflammation is recognized by granulomas, or meningeal, perivascular space, or lumbar nerve roots enhancement . VasculaOne of the most important differentials for neurobrucellosis in TB endemic areas is neurotuberculosis . In factNeurobrucellosis is a diagnostic challenge in Brucella endemic areas. The symptoms could be mild and nonspecific and the classic triad of meningitis is uncommon. Behavior changes are among the important clinical features are neurobrucellosis. Differentiation between neurobrucellosis and systemic brucellosis is important, because more prolonged treatment is indicated for neurobrucellosis and it could be associated with a wide spectrum of complications that require close neurologic monitoring.Mild CSF pleocytosis of fewer than 50 leukocytes per microliter of CSF was common and identified in about half of the cases of neurobrucellosis. Nevertheless, severe hyperproteinorrhachia and severe hypoglycorrhachia were rare in neurobrucellosis. The yield of Brucella culture from CSF and blood is low in cases of neurobrucellosis and the culture-confirmed diagnosis was made in less than 10% of patients.. When a very high CSF protein level is among the known features of neurotuberculosis, it is rare in neurobrucellosis and should raise the possibility of neurotuberculosis as a more probable diagnosis.An important differential for neurobrucellosis in TB endemic areas is neurotuberculosis


Apoe−/−) mice. Herein, we investigated whether Brussels chicory can stabilize advanced plaques in the aorta via improving oxidative stress. Thirty week old Apoe−/− mice were fed the AIN-93G diet or supplemented with 0.5% freeze-dried Brussels chicory for twenty weeks. Aortic plaque size and stability, aortic relaxation, monocyte adhesion to aortic endothelium, free radicals, and enzymatic and non-enzymatic factors involved in free radical production and elimination in aorta and serum were measured. Brussels chicory consumption did not alter aortic plaque size, however, it stabilized aortic plaques, promoted aortic relaxation, and also inhibited monocyte adhesion to aortic endothelium. Moreover, this administration reduced oxidized LDL (ox-LDL) and 4-hydroxynonenal (4-HNE) content in aortic plaques, associated with inhibited aortic NADPH oxidase (NOX) and uncoupled endothelial nitric oxide synthase (eNOS)-mediated free radical production. However, Brussels chicory consumption did not appreciably alter aortic and serum superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities, aortic glutathione (GSH), as well as serum non-enzymatic antioxidants, such as bilirubin, uric acid, and GSH. Collectively, improved oxidative stress might contribute to the atheroprotective effect of Brussels chicory, supporting the prospect of the antioxidant therapy in advanced atherosclerosis progression.Brussels chicory, a typical vegetable in Mediterranean diets, has been recently reported to stabilize advanced atherosclerotic plaques in the brachiocephalic artery of apoE-deficient ( Cichorium intybus L. var. foliosum, Belgian endive), a typical Mediterranean vegetable, could stabilize advanced atherosclerotic plaques in the brachiocephalic artery of a apolipoprotein E-deficient (Apoe−/−) mouse model [Cardiovascular diseases are the leading global cause of death, with atherosclerosis being a major contributor . Adherense model . As athese model , it remaPersistent oxidative stress is vital for the etiology and progression of atherosclerosis . OxidatiCichorium intybus L., Asteraceae) has a long-standing medicinal use dating back to the ancient Egyptian times [Apoe−/− mice [Chicory  with advanced atherosclerotic plaques [The procedures of mouse studies and mouse tissue samples were adopted from our previous studies . Thirty  plaques  were fedAdvanced plaque size and stability were assessed by either hematoxylin and eosin or Sirius red staining as we previously described . Serial Cellular and molecular compositions of advanced plaques were analyzed using immunohistochemistry of frozen sections of heart as we previously documented . PrimaryAortic relaxation was measured with acetylcholine  or sodium nitroprusside  as we previously described . A 3 mm 6 calcein-acetoxymethyl-ester-labeled HL-60 monocytes for 30 min. After incubation, unbound HL-60 cells were removed by ice-cold PBS. Images were captured with Leica TCS SP5 laser scanning confocal microscope by two observers blind to the experimental groups, and the number of adherent monocytes were then counted in 5 consistent fields. The ex vivo adhesion of monocyte to the ascending aortic endothelium was evaluated as we previously described . In brieOxidative stress in advanced plaques was evaluated by fluorescent dihydroethidium (DHE) staining as previously described . Frozen Ox-LDL and 4-HNE in advanced plaques of frozen tissue sections were analyzed using immunohistochemistry assays. Primary antibodies used were rabbit polyclonal anti-ox-LDL  and mouse monoclonal anti-4-HNE . The omission of primary antibodies was used as negative controls. Images of the stained samples were captured by an operator blinded to the experimental groups. The area occupied by a particular stain was shown as a percentage of plaque size. Superoxide levels in thoracic and abdominal aorta rings were assessed by lucigenin-enhanced chemiluminescence as previously documented . SamplesThe eNOS activity was determined using eNOS assay kit , following the manufacturer’s instructions. The XO activity in aortas were measured by pterin-based method . In brie2O2. The GPx activity was evaluated using an NADPH decay and expressed as units/mg protein in aortas or units/mL in serum. The CAT activity was assayed using the protocol of Aebi [2O2 decomposition. In brief, the protein samples were added to phosphate buffer containing H2O2 and the consumption of H2O2 was then spectrophotometrically measured at 240 nm.The SOD activity was determined using WST-1 method with commercial kit, obtained from Nanjing Jiancheng Bioengineering Institute (A001-3), according to the manufacturer’s protocol. The GPx activity was assessed with spectrophotometry as previously described . Briefly of Aebi  by monitTotal RNA was isolated from aortas using TRI reagent . RNA concentration measurement was based on the absorbance at 260 nm. For mRNA quantification, RNA was reverse-transcribed to cDNA using QuantiTect Reverse Transcription Kit , followed by quantitative real-time PCR with SYBR Green PCR Master Mix  performed on ABI 7300 Real-Time qPCR System. β-actin was used as an internal control. The primer sequences of indicated genes were listed in The GSH and oxidized GSH (GSSG) level in aorta and serum were measured by a commercial kit . The albumin and uric acid level in serum was assayed using mice serum albumin ELISA kit  and uric acid assay kit , respectively. The vitamin C, vitamin E, and total bilirubin level in serum were assayed using corresponding colorimetric assay kit . p < 0.05.Data were expressed as the means ± SD. Normality of the data were tested by the Kolmogorov–Smirnov test. The data were analyzed by a two-tailed unpaired Student’s t-test with or without the Welch correction performed on GraphPad Prism Version 9.3.1. Statistical significance was accepted for Morphometric analyses show that Brussels chicory consumption for 20 weeks does not significantly affect aortic root plaque size as compared with the control treatment A,B. Instp < 0.05; + (a common marker for macrophages) and ACTA2+ (a common marker for vascular smooth muscle cells) cells  cells B,C in aoBrussels chicory consumption promotes endothelium-dependent relaxation to Ach A. This e·− levels assessed by DHE staining (p < 0.05).Compared with the control treatment, Brussels chicory consumption elicits a significant reduction in oxidative stress in aortic root plaques, as indicated by a reduction in O2staining A. Concorstaining B and 4-Hstaining C in aort·− in thoracic and abdominal aortas, whereas apocynin (a specific inhibitor for NOX) but not L-NAME (a specific inhibitor for eNOS) and oxypurinol (a specific inhibitor for XO) abolish the inhibitory effect of Brussels chicory on thoracic and abdominal aortic O2·− production  in aortic plaques prompted us to dissect the underlying mechanisms. Brussels chicory does not affect the expression and activities of aortic antioxidant enzymes SOD, CAT, or GPx, or the content of reduced GSH, an important endogenous non-enzymatic antioxidant. These observations strongly suggest that the oxidative stress-modulation effect of Brussels chicory inside aortic plaques is not due to enhanced free radical elimination. It should be pointed out that other antioxidant factors in aortic plaques, such as Nrf2  and thiostration . These fApoe−/− mice with advanced atherosclerosis [Increased free radical production and reduced antioxidant GSH bioavailability were demonstrated in experimental and human atherosclerosis ,47. Pharclerosis . Our curclerosis ,39, bothclerosis ,52. Apoe−/− mice. Together with other works [In conclusion, we showed that Brussels chicory can increase plaque stability in the aorta, at least in part through improving oxidative stress within plaques by inhibition of NOX and uncoupled eNOS-mediated free radical production in male er works ,39,40,41