ADAMTS10 have long been known to cause autosomal recessive Weill-Marchesani Syndrome which is characterized by short stature and ocular abnormalities, more recent work has shown that certain mutations in ADAMTS10 cause glaucoma in dogs. In humans, glaucoma is the leading cause of irreversible vision loss that affects tens of millions of people world-wide. Vision loss in glaucoma is a result of neurodegeneration of retinal ganglion cells that form the inner-most layer of the retina and whose axons form the optic nerve which relays visual information to the brain. ADAMTS10 contributes to the formation of microfibrils which sequester latent transforming growth factor β (TGFβ). Among its many biological functions, TGFβ promotes the development of retinal ganglion cells and is also known to play other roles in glaucoma pathogenesis. The aim of this study was to test the hypothesis that ADAMTS10 plays a role in retinal ganglion cell development through regulation of TGFβ signaling. To this end, Adamts10 expression was targeted for reduction in zebrafish embryos carrying either a fluorescent reporter that labels retinal ganglion cells, or a fluorescent reporter of pSmad3-mediated TGFβ family signaling. Loss of adamts10 function in zebrafish embryos reduced retinal ganglion cell reporter fluorescence and prevented formation of an ordered retinal ganglion cell layer. Targeting adamts10 expression also drastically reduced constitutive TGFβ signaling in the eye. Direct inhibition of the TGFβ receptor reduced retinal ganglion cell reporter fluorescence similar to the effect of targeting adamts10 expression. These findings unveil a previously unknown role for Adamts10 in retinal ganglion cell development and suggest that the developmental role of Adamts10 is mediated by active TGFβ family signaling. In addition, our results show for the first time that Adamts10 is necessary for pSmad3-mediated constitutive TGFβ family signaling.Although mutations in  ADAMTS10 as disease-causative for a colony of Beagle dogs with autosomal recessive inheritance of glaucoma , a rare connective tissue disorder characterized by short stature and ocular abnormalities including glaucoma (FBN1)  , suggesta (FBN1) . FBN1 asa (FBN1) . Microfia (FBN1) . Mutatioa (FBN1) . ADAMTS1a (FBN1) . Recent activity . Mutatioactivity . Therefoactivity .adamts10 mRNA has been shown in the developing zebrafish embryo (ADAMTS10 is abundantly expressed in the developing mouse eye and up-regulation of h embryo . In the Danio rerio) were maintained in aquatic housing units on a monitored recirculating system at 26°C with standard light/dark (14/10 h) cycle. Experiments were performed using embryos from outcrosses of wildtype AB strain to a transgenic reporter line that expresses GFP under control of a pou4f1 (brn3a) enhancer element, Tg (pou4f1-hsp70l:GFP)rw0110bTg formerly Tg (brn3a-hsp70l:GFP)rw0110bTg provided by Dr. Takanori Hamaoka and Dr. Hitoshi Okamoto of the Riken Brain Institute through the European Zebrafish Resource Center (Tg(12xSBE:EGFP)ia16Tg, generated by Dr. Francesco Argenton, Universita di Padova, obtained through the European Zebrafish Resource Center  following the manufacturer’s protocol. The adamts10 probe set was checked to avoid cross detection of the adamts6 which has high homology with adamts10. A probe set for DapB, a bacterial gene, was used as negative control. Briefly, embryo sections were post-fixed in 4% PFA/PBS for 15 min at 4°C, dehydrated through gradient ethanol then treated with hydrogen peroxide for 10 min at room temperature. Target retrieval was conducted by submerging slides in retrieval solution for 5 min at 100°C. Following 30 min protease plus treatment at 40°C in a hybridization oven , samples were hybridized with probes, subjected to signal amplification steps and then reacted with Fast Red. Bright field images were captured using a Nikon Eclipse microscope equipped with a ×40 objective.Zebrafish embryos or adult eyes were embedded in 20% sucrose/OCT and 7 μm-thick cryosections were made as described below for immunohistochemistry. RNA echnique  using a Injections of 1–2 nl were made into the yoke adjacent to the embryo at the one-cell stage at approximately 12:00 p.m. To visualize injections, 0.025% phenol red was included in the injection solution. After injection, embryos were incubated at 29°C in egg water . At approximately 10:00 a.m. on post-injection days 1, 2 or 3 , embryos were euthanized in 300 mg/L tricaine methanesulfonate, then fixed in 4% PFA/PBS. For fluorescence microscopy, 0.2 mM phenylthiourea (PTU) was added post-injection to prevent pigmentation.adamts10 mRNA (referred to from here on as adamts10 MO) was injected at 4–5 ng/embryo. Human ADAMTS10 mRNA was transcribed from expression plasmids described previously (in vitro T7 ultra mMessage mMachine™ kit with poly-A tail addition (Thermo Fisher Scientific). For MO rescue experiments, embryos were co-injected with 4 ng adamts10 MO with 400 ng of either normal or G661R mutated human ADAMTS10 mRNA.MO obtained from Gene Tools , were reconstituted to 1 mM with RNase/DNase-free water and stored at room temperature as suggested by the manufacturer. Translation-blocking MO (5′-CCAAACTCCTCCACACCGTTTCCAT-3′) targeting the translation initiation complex of eviously  using thTg(12xSBE:EGFP) embryos within 8 h after treatment (Tg (pou4f1-hsp70l:GFP) or Tg(12xSBE:EGFP) embryos. This treatment protocol did not induce gross disruption of eye morphology, though treated embryos were slightly smaller than untreated , a concentration previously shown to drastically reduce reporter fluorescence of reatment , or vehintreated . At 48 hGFP-positive embryos were dechorionated, fixed in 4% PFA/PBS for 2 h at room temperature or at 4°C overnight, then stored in PBS at 4°C. For imaging, embryos were placed ventral side up in 3% (w/v) methyl cellulose and fluorescence and brightfield images taken with a ×4 objective with a Nikon AZ100M fluorescence microscope.Embryos were embedded in 20% sucrose in optimal cutting temperature compound (OCT) as described by For immunostaining, cryosections from the center of the eye were rehydrated with PBS before addition of blocking buffer  for 1 h. Blocking buffer was removed and primary antibody solution in incubation buffer  was added. The antibodies used were 1:1,000 rabbit anti-GFP (Torrey Pines Biolabs) and 1:25 mouse anti-Isl1 . After overnight incubation at 4°C in a humidified chamber, sections were washed 3 times in PBS for 10 min and incubated in 1:1,000 Alexa Fluor-donkey-anti-mouse-546 and 1:1,000 Alexa Fluor-donkey-anti-rabbit-488 (Invitrogen) in secondary incubation buffer  for 1 h. Sections were then washed 3 times with PBS and mounted with Prolong Gold mounting fluid with DAPI (Invitrogen).http://rsb.info.nih.gov/ij/plugins/surface-plot-3d.html). Statistical analyses were performed in Graphpad Prism. Statistical tests and sample sizes are indicated in the figure legends.Integrated fluorescence density of fluorescent embryo images, defined as the product of the area of the region of interest (ROI) and the mean gray value, was measured by drawing an ROI around the retina using the ImageJ polygon tool. 3D surface plots were generated from single-channel GFP images using the NIH ImageJ interactive 3D surface plot plug-in (adamts10 mRNA in the developing zebrafish embryos and in the adult zebrafish eye has been shown in a previous RT-PCR study (in situ hybridization (Advanced Cell Diagnostics), we found adamts10 mRNA expression in the eye of zebrafish embryos at 24, 48, and 72 hpf  line of zebrafish, which expresses GFP driven by a pou4f1 enhancer element  is specifically expressed in ∼80% of developing and differentiated RGCs . We used element  to test rescence . In uninrescence . As showrescence . Howeverc nerves . At 48 hadamts10, rescue experiments were performed in which Tg(pou4f1-hsp70l:GFP) embryos were co-injected with adamts10 MO and human ADAMTS10 mRNA (which is not targeted by the zebrafish adamts10 MO). GFP fluorescence was compared at 48 hpf to embryos that were uninjected or injected with adamts10 MO only. In uninjected embryos, the retina displayed strong fluorescence with staining of the optic nerve consistent with labeling of RGCs , with a more uniform distribution of fluorescence intensity along the retina  in integrated fluorescence density  embryos using an antibody against GFP and an antibody against Isl1, which is expressed in postmitotic RGCs and is required for RGC development  line of zebrafish which expresses GFP in response to activation of a pSmad3 binding element, thereby acting as a reporter of active TGFβ superfamily signaling (Tg(12xSBE:EGFP) embryos (adamts10 MO resulted in a 66.6% reduction in fluorescence as compared to uninjected (p < 0.0001). These results indicate that targeting Adamts10 expression drastically reduces constitutive pSmad3-mediated TGFβ superfamily signaling in the retina.ADAMTS10 plays a role in microfibril structure and function . Since mignaling . In unin embryos , indicatadamts10 MO suggested that Adamts10 may exert its effect on RGC development through suppression of TGFβ signaling. To test this hypothesis, embryos were treated with an inhibitor of the TGFβ receptor, SB431542, at 11 hpf, a time at which the optic primordium has just formed (Tg(12xSBE:EGFP) line treated with SB431542 showed nearly complete reduction of pSmad3-driven GFP fluorescence, verifying effective inhibition of TGFβ signaling, while vehicle control (DMSO) had no effect (Tg(pou4f1-hsp70l:GFP) embryos were treated with SB431542. Treatment with SB431542, but not vehicle control, strongly reduced pou4f1 enhancer-driven GFP expression  which is expressed early in post-mitotic RGC precursors and plays an important role in their development (Tg(pou4f1-hsp70l:GFP) line of zebrafish that express GFP driven by a pou4f1 enhancer element  embryos using antibodies against GFP and Isl1 which is expressed in post-mitotic RGCs and plays key roles in RGC development  line of zebrafish that expresses GFP in response to activation of a pSmad3 binding element. We found that uninjected Tg(12xSBE:EGFP) embryos expressed abundant GFP throughout the retina at 48 hpf  signaling in mouse embryo fibroblasts (ADAMTS10 deficiency could vary between tissues, developmental stages and species investigated. Another signaling role for ADAMTS10 described by Cain et al. that we did not investigate is its role in formation of focal adhesions and epithelial cell-cell junctions (A role for ADAMTS10 in TGFβ family signaling contrasts with the results of Mularczyk et al., who found that a premature termination mutation of roblasts . Althougunctions .adamts10 MO (via a pSmad3-mediated TGFβ family pathway.TGFβ is involved in neuronal development, including in the retina, in programmed cell death and in axon specification . This lemts10 MO . Howevermts10 MO . Althougmts10 MO . Our resadamts10 expression. This indicates that the primary defect of RGC development in adamts10 MO treated embryos is likely a failure to migrate to the appropriate position resulting in a retinal lamination defect.Isl1 and Pou4f1 are expressed in early post-mitotic RGCs and are known to play important roles in their differentiation . ExpressIn summary, we have discovered a previously unknown role for Adamts10 in RGC development, possibly contributing to their apical to basal translocation and formation of an ordered ganglion cell layer. Our results suggest that the developmental role of Adamts10 is mediated by active TGFβ family signaling. In addition, our results show for the first time that Adamts10 is necessary for pSmad3-mediated constitutive TGFβ family signaling in the developing retina.


The endogenous hypercortisolism that characterizes Cushing’s syndrome (CS) is associated with a state of hypercoagulability that significantly increases the risk of thromboembolic disease, especially, venous events. Despite this certainty, there is no consensus on the best thromboprophylaxis strategy (TPS) for these patients. Our aim was to summarize the published data about different thromboprophylaxis strategies, and to review available clinical tools assisting thromboprophylaxis decision making.Narrative review of thromboprophylaxis strategies in patients with Cushing’s syndrome. A search was carried out on PubMed, Scopus and EBSCO until November 14th, 2022, and articles were selected based on their relevance and excluded in case of redundant content.Literature is scarce regarding thromboprophylaxis strategies to be adopted in the context of endogenous hypercortisolism, most often being a case-by-case decision according to the centre expertise. Only three retrospective studies, with a small number of patients enrolled, evaluated the use of hypocoagulation for the thromboprophylaxis of patients with CS in the post-operative period of transsphenoidal surgery and/or adrenalectomy, but all of them with favourable results. The use of low molecular weight heparin is the most frequent option as TPS in CS context. There are numerous venous thromboembolism risk assessment scores validated for different medical purposes, but just one specifically developed for CS, that must be validated to ensure solid recommendations in this context. The use of preoperative medical therapy is not routinely recommended to decrease the risk of postoperative venous thromboembolic events. The peak of venous thromboembolic events occurs in the first three months post-surgery.The need to hypocoagulate CS patients, mainly in the post-operative period of a transsphenoidal surgery or an adrenalectomy, is undoubtable, especially in patients with an elevated risk of venous thromboembolic events, but the precise duration and the hypocoagulation regimen to institute is yet to be determined with prospective studies. Cushing’s syndrome (CS) is associated with an eighteen-fold higher risk of venous thromboembolic events (VTE) compared with the general population . Indeed,This hypercoagulable state is consequence of both quantitative and qualitative alterations in the haemostatic system induced by the cortisol excess.In one hand, by the increase in plasma clotting factors - especially factor VIII and Von Willebrand factor (VWF), but also factors IX, X and XI -, the decrease in plasma tissue factor pathway inhibitor (TFPI) and the impairment of fibrinolysis – by the upregulation of the synthesis of plasminogen activator inhibitor type I (PAI-1). On another hand, the overexpression of abnormally high molecular weight VWF multimers capable of inducing spontaneous platelet aggregation contribute to a higher risk of thrombotic events.Coagulation profiles in patients with CS are heterogeneously affected. The hemostatic abnormalities most consistently reported are shortening of activated partial thromboplastin time (aPTT) and increased thrombin generation –22. Intep = 0.007)  OR “Cushing disease” [MeSH Terms] OR “hypercortisolism” [MeSH Term] AND “coagulation” [MeSH Term] OR “hypercoagulability” [MeSH Terms]. Secondly, with the intention to find articles that could bring us some light about different thromboprophylaxis strategies implemented in these patients, and also their benefits and risks, with carried out the research with the terms “Cushing syndrome” [MeSH Terms] OR “Cushing disease” [MeSH Terms] OR “hypercortisolism” [MeSH Term] AND “anticoagulant agents” [MeSH Terms].Only three retrospective studies Table , with a p < 0.001), without any significant haemorrhagic event in the group of patients that received hypocoagulation strategy.The first one was done by Boscaro M et al. in 2002 . They cop = 0.081). No bleeding complications were observed during the follow-up in either group.The second one was conducted in 2015 by Barbot M et al. . LM. LM44]. insights . So, basinsights .It is also important to mention that this pharmacologic thromboprophylaxis should be used in addition to the mechanical thromboprophylaxis, e.g., intermittent pneumatic compression stockings or elastic stockings, to augment the efficacy of the thromboprophylaxis strategy.With respect to the use of PMT to reduce the risk of post-operative VTE, given its unclear benefit and potential adverse effects, it is not routinely recommended. Nevertheless, it should be offered on an individual basis, especially if surgery is delayed or if hypercortisolism is markedly severe .In conclusion, thromboprophylaxis strategy is not about a single, simple decision to initiate or not hypocoagulation drugs. It is all about the continuous balancing of risks and benefits, which must always be individualized, in the pre and postoperative period, for each patient who presents with CS. Nevertheless, based on the literature reviewed here, we can state a general recommendation about the adequate strategy of thromboprophylaxis to implement in patients with CS Fig. .


This review systematically evaluated radiomics analysis procedures for characterizing salivary gland tumors (SGTs) on magnetic resonance imaging (MRI). Radiomics analysis showed potential for characterizing SGTs on MRI, but its clinical application is limited due to complex procedures and a lack of standardized methods. This review summarized radiomics analysis procedures, focusing on reported methodologies and performances, and proposed potential standards for the procedures for radiomics analysis, which may benefit further developments of radiomics analysis in characterizing SGTs on MRI.Radiomics analysis can potentially characterize salivary gland tumors (SGTs) on magnetic resonance imaging (MRI). The procedures for radiomics analysis were various, and no consistent performances were reported. This review evaluated the methodologies and performances of studies using radiomics analysis to characterize SGTs on MRI. We systematically reviewed studies published until July 2023, which employed radiomics analysis to characterize SGTs on MRI. In total, 14 of 98 studies were eligible. Each study examined 23–334 benign and 8–56 malignant SGTs. Least absolute shrinkage and selection operator (LASSO) was the most common feature selection method (in eight studies). Eleven studies confirmed the stability of selected features using cross-validation or bootstrap. Nine classifiers were used to build models that achieved area under the curves (AUCs) of 0.74 to 1.00 for characterizing benign and malignant SGTs and 0.80 to 0.96 for characterizing pleomorphic adenomas and Warthin’s tumors. Performances were validated using cross-validation, internal, and external datasets in four, six, and two studies, respectively. No single feature consistently appeared in the final models across the studies. No standardized procedure was used for radiomics analysis in characterizing SGTs on MRIs, and various models were proposed. The need for a standard procedure for radiomics analysis is emphasized. Salivary gland tumors (SGTs) constitute approximately 2–6.5% of all head and neck tumors, with about 80% originating from the parotid gland ,2,3. TheMagnetic resonance imaging (MRI) is widely employed in mapping the SGTs for treatment plans because it provides detailed information on soft tissue. MRI has also demonstrated comparable efficacy to FNAC in characterizing SGTs ,10,11. MThis systematic review aims to evaluate studies that have assessed the performance of radiomics analysis in characterizing benign and malignant SGTs or PA and WT on MRI. The primary focus is analyzing the methodologies and performances reported in each eligible study to provide a comprehensive summary of the approaches employed during the radiomics analysis procedure. The aim is to contribute to the standardization of a radiomics analysis procedure for characterizing SGTs on MRI, which may facilitate its translation into clinical practice.The analysis and inclusion criteria methods were pre-defined and documented for this systematic review. The review protocol was registered at PROSPERO International Prospective Register of Systematic Reviews (ID: CRD 42023446728). This review followed the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) 2020 statement [A systematic literature search was conducted through PubMed, Embase, Web of Science, Scopus, and Cochrane Library. The search encompassed studies published from the inception of the electronic databases up to 20 July 2023. The search terms used were “ AND radiomics AND (MRI OR magnetic resonance imaging)”. These terms were chosen to ensure the inclusion of all salivary gland tumors and to summarize the methodologies and diagnostic performance of radiomics analysis in characterizing SGTs on MRI. The search results were stored in an Excel spreadsheet . Duplicate titles were screened and removed. As the data for this review were obtained solely from previously published studies, Institutional Review Board approval or written patient consent was not required and, therefore, waived.Original articles published in English.Participants: studies involving patients with SGTs who underwent pre-treatment head and neck MRI scans, including at least T1-weighted (T1W), T2-weighted (T2W), contrast-enhanced T1W (CE-T1W), CE-T2W, or diffusion-weighted imaging (DWI).Comparison: studies reporting the performance of radiomics analysis in characterizing SGTs on MRI.Outcomes: the primary outcome was the performance of radiomics analysis in characterizing benign and malignant SGTs on MRI; the second outcome was the performance of radiomics analysis in characterizing PA and WT on MRI.The inclusion criteria were as follows:Articles in the form of reviews, guidelines, conference proceedings, or case reports/series.Studies that did not report the area under the curve (AUC) of the radiomics models in characterizing SGTs.Studies with patient populations overlapped with previous studies conducted in the same investigated institution for assessing the same outcomes. The exclusion criteria were based on the publication time, with later studies being excluded.The exclusion criteria were as follows:Two observers (QYHA and KFH) independently screened the records based on the title and abstract, and full-text evaluation was performed for selected records. Any disagreements were resolved through discussion or consultation with a third observer (TYS).Study characteristics: first author, journal name, year of publication, city, patient recruitment period, and study design (prospective or retrospective).Patient characteristics: number of patients in the training, testing, and external datasets, methods for diagnosis of the nature of salivary gland tumors.MRI characteristics: MRI sequences used for analysis.Radiomics analysis procedure: segmentation method, number of features extracted, feature categories, methods for feature selection, number and categories of the selected features, names of the selected features, classifiers used for model build-up, and final model.Outcomes: model performance in training, testing, and external datasets.One observer (KM) extracted the following data from the included studies:A second observer (QYHA) verified the received data in an Excel spreadsheet.All of the eligible studies underwent assessments of study quality by one observer (KM) using the Quality Assessment of Diagnostic Accuracy Studies-2 (QUADAS-2) tool and The Radiomics Quality Score (RQS). The QUADAS-2 evaluates four domains: patient selection, index test, reference standard, and flow and timing . Each doThe inter-observer agreements for article selection based on titles and abstracts were calculated using Cohen’s kappa coefficients.Inter-observer agreement for the extracted features was assessed in 7/14 studies (50%), with all studies using an intra-class correlation coefficient (ICC) threshold of >0.75 to indicate high repeatability. The most common method for feature selection was the least absolute shrinkage and selection operator (LASSO), used in 8/14 studies (57.14%), followed by analysis of variance (ANOVA) in 3/14 studies (21.43%). Cross-validation or bootstrap techniques were applied in 11/14 studies (78.57%) to enhance the stability of the selected features. Logistic regression was the most commonly employed classifier for building the radiomics model, used in 7/14 studies (50%), followed by support vector machine (SVM) in 6/14 studies (42.86%).Nine studies reported the final selected features for characterizing benign and malignant SGTs. The number of features selected for building the final model ranged from 3 to 17, a total of 61 features . Four studies reported the final selected features for characterizing PA and WT. The number of features selected for building the final model ranged from 4 to 13, a total of 36 features . No features included in the final models were found to be present in more than two studies.Thirteen studies reported final radiomics models . The perFor characterizing benign and malignant tumors, the AUCs ranged from 0.74 to 1, of which studies that used only T1W or T2W images achieved AUCs ranging from 0.74 to 0.85, and those that used only DWI images achieved AUCs ranging from 0.76 to 0.89. The highest AUC was achieved using features selected from multi-parametric MRI  validated on a cross-validation dataset, while the lowest AUC was obtained using T2WI.Three studies compared the performance of radiomics models built using different classifiers. One study reported that radiomics models constructed with support vector machine (SVM) (AUC 0.893) and logistic regression (LR) (AUC 0.886) outperformed those built with k-nearest neighbors (KNN) (AUC 0.796) . AnotherTwo studies suggested that models incorporating clinical and radiomics features exhibited superior performance compared to models built solely with clinical or radiomics features ,20.Three studies compared the performances of radiomics models constructed using different MRI sequences. It was observed that models utilizing multiple MRI sequences for feature extraction outperformed those utilizing a single sequence ,21,23. AOne study demonstrated that a radiomics model constructed using a single feature category outperformed a model utilizing all feature categories . AnotherFor characterizing PA and WT, the AUCs ranged from 0.80 to 0.96, of which studies that used only T1W or T2W images achieved AUCs of 0.80–0.96, and those that used only DWI images achieved an AUC of 0.93. The highest and lowest AUCs were achieved using features selected from T2W images.Similar to the BT vs. MT studies, three studies indicated that models incorporating clinical and radiomics features outperformed those constructed using only clinical or radiomics features ,22,25. OThe initial flow of QUADAS-2 assessments is presented in The median score of the RQS was 12.5 out of 36, with a range of 5 to 16 . A totalIn this systematic review, we evaluated the procedures for radiomics analysis in characterizing SGTs on MRI based on 14 eligible studies. Ten studies focused on characterizing benign and malignant tumors, reporting AUC values ranging from 0.74 to 1. Seven studies differentiated PA and WT, with AUCs ranging from 0.80 to 0.96. Despite the promising accuracy of radiomics, with most studies achieving AUCs above 0.80, these studies employed various methods for radiomics analysis, and none of the resulting models have been further validated or widely implemented in clinical practice. To note, no features included in the final models were found to be present in more than two studies. This systematic review provides valuable insights into MRI sequence selection, image preprocessing, feature extraction, feature selection, and model development, which can help standardize the procedures for radiomics analysis in characterizing SGTs on MRI.The MRI protocol for SGTs typically includes multiple MRI sequences, such as T1WI, T2WI, and CE images. Functional MRI techniques, such as dynamic contrast-enhanced (DCE) MRI and DWI, have also demonstrated potential for characterizing SGTs and are increasingly included in the MRI protocol for SGTs . In radiImage preprocessing and feature extraction play crucial roles in radiomics analysis, but their implementation and reporting were inconsistent across the studies reviewed. Only five studies reported on image preprocessing, while the details of this step were unclear in the remaining studies. Image preprocessing is essential for standardizing images obtained from different institutions with varying protocols. This step is particularly critical but challenging to implement for MRI as MRIs are constructed by weighting the signals corresponding to the magnetic properties of the tissues being imaged. Several common methods have been proposed to address different protocol parameters and improve the quality and consistency of MRI images, including the “μ ± 3σ” method, N4ITK bias field correction, resampling, and Z-score normalization ,37,38,39The selection of ROI is an important consideration and depends heavily on the heterogeneity of the tumor as depicted on imaging. Since SGTs can exhibit high heterogeneity, it is suggested to use an ROI covering the whole tumor. Interestingly, one study showed that radiomics models built using features from manual segmentation outperformed those using automatic segmentation for characterizing benign and malignant SGTs on MRI . This maOn the other hand, in diseases with low incidences, such as SGTs, the number of features extracted for analysis should be carefully considered. The reviewed studies had total patient numbers ranging from 31 to 334, and the total number of extracted features ranged from 29 to 3396. Most studies extracted over 400 features. These increased the risk of overfitting the model. One study notably demonstrated that the radiomics model built up by the selection from over 1000 radiomics features did not outperform that from 91 features . FurtherThe review showed the LASSO method as the most commonly used feature selection method in the analyzed studies, with 8/14 studies employing it. LASSO is a regularization-based method that can effectively remove irrelevant or redundant features by shrinking their coefficients to zero . This heLR and SVM were the most commonly used classifiers for building radiomics models to characterize SGTs on MRI. However, the best-performing classifier varied among the studies. In three studies that compared multiple classifiers, SVM and LR outperformed KNN in one study, XGBoost and SVM performed better than DT in another study, and SVM and LDA performed similarly in the third study ,18,21. MFocus on non-contrast-enhanced MRIs.Implement image preprocessing.Limit the number of features extracted and consider the feature categories.Evaluate inter-observer agreement for the extracted features and select those with high repeatability for further analysis.Use multiple feature selection methods.Ensure feature stability by different approaches.Build models using different approaches and identify the best model.Validate models using at least cross-validation dataset with/without internal or external datasets.Report the final models for future validations.Open datasets  for future validations.Based on the findings of this review, the following suggestions can be made to facilitate radiomics analysis and the development of radiomics models in characterizing SGTs on MRI:Implementing these recommendations will enhance the standardization, reproducibility, and generalizability of radiomics analysis and radiomics models in characterizing SGTs on MRI, ultimately facilitating their broader use in clinical practice.This review has limitations that may result in inherent heterogeneity and publication bias. Firstly, all studies included in this systematic review were retrospective studies. Due to the retrospective nature, the concerns regarding the risk of bias in the included studies could not be avoided. However, the results from QUADAS-2 showed that most of the studies had a low risk of bias and high applicability. Secondly, a meta-analysis that evaluated the performance of radiomics models in characterizing SGTs on MRI was inappropriate due to the significant heterogeneity of the radiomics models among the eligible studies. Thirdly, according to the registered study protocol, eligible studies that were published after July were not included in the analysis. Furthermore, the added value of radiomics analysis to clinical practice remains underreported, and no studies have analyzed cost-effectiveness. Therefore, it is recommended that further studies be conducted to evaluate radiomics analysis using standardized procedures for characterizing SGTs on MRIs.Previous studies have demonstrated the potential of radiomics analysis in characterizing SGTs on MRI. However, the lack of standardized procedures for implementing radiomics analysis across these studies may have led to the limited generalizability of the final radiomics models, thereby restricting their application in clinical practice. To develop radiomics models that can be widely utilized for characterizing SGTs on MRI in the future, it is crucial to establish a consensus on the procedures for conducting radiomics analysis.


Yersinia pestis, is a zoonotic disease that can reemerge and cause outbreaks following decades of latency in natural plague foci. However, the genetic diversity and spread pattern of Y. pestis during these epidemic-silent cycles remain unclear. In this study, we analyze 356 Y. pestis genomes isolated between 1952 and 2016 in the Yunnan Rattus tanezumi plague focus, China, covering two epidemic-silent cycles. Through high-resolution genomic epidemiological analysis, we find that 96% of Y. pestis genomes belong to phylogroup 1.ORI2 and are subdivided into two sister clades (Sublineage1 and Sublineage2) characterized by different temporal-spatial distributions and genetic diversity. Most of the Sublineage1 strains are isolated from the first epidemic-silent cycle, while Sublineage2 strains are predominantly from the second cycle and revealing a west to east spread. The two sister clades evolved in parallel from a common ancestor and independently lead to two separate epidemics, confirming that the pathogen responsible for the second epidemic following the silent interval is not a descendant of the causative strain of the first epidemic. Our results provide a mechanism for defining epidemic-silent cycles in natural plague foci, which is valuable in the prevention and control of future plague outbreaks.Plague, caused by  Yersinia pestis isolates spanning more than 60 years and covering two epidemics and "silent” cycles in-between reveal the phylogeographic and evolutionary background of these plague cycles.Genome sequencing of Chinese  Yersinia pestis1. Three historic plague pandemics have caused over 160 million deaths worldwide2. The most recent of these pandemics, which began at the end of the 19th century in Hong Kong, China, and then spread to Africa, America, Oceania, and other parts of the world via maritime trade, lasted until the mid-20th century5. At present, plague remains a threat in many parts of the world and has been categorized by the World Health Organization since 2000 as a reemerging disease8.Plague is a deadly infectious disease caused by the gram-negative bacterium Y. pestis can shape the natural plague foci under suitable ecological conditions, sylvatic plagues in these foci exhibit recurrent cycles of epidemic and silent periods with intervals ranging from several years to decades, rather than a single continuous epidemic10. During the epidemic period, animal plague usually occurs prior to human plague, whereas in the silent period, human plague disappears and animal plague fades out with occasional rodents detected carrying the F1 antibody or antigen around the corresponding foci12. However, there is a lack of knowledge regarding the genomic diversity of Y. pestis during these epidemic-silent intervals. It is unknown whether the reactivation of plague foci is caused by invasion of a new Y. pestis strain from other populations or the awakening of offspring from the latent strain.Although Rattus tanezumi plague focus  is one of the 12 major natural plague foci in China13. Active animal plague surveillance in this plague focus has been conducted since 195114, and two epidemic periods and two silent periods have been documented since 195016. The first plague epidemic period (named Epidemic1) was observed from 1950 to 1956, during which a plague outbreak occurred in 12 counties/cities in western Yunnan, involving 2950 human cases and 633 deaths16. Since 1957, plague has been well controlled, with few Y. pestis strains, bacteriophages, and positive sera of animals observed within the investigation area17. However, after a silence of 25 years, which we named the Silent1 period (1957–1981), another epidemic wave (Epidemic2) emerged in animals and subsequently spread to humans, lasting until 2007. Since 2008, cases of plague have faded again and the natural plague focus has entered a new silent period (Silent2), which is ongoing to present18.The Yunnan Y. pestis isolates in Yunnan provide us with an opportunity to investigate the dynamics of the pathogen’s genomic diversity during these epidemic-silent cycles. Here, we analyzed the whole genomes of 356 Y. pestis strains isolated between 1952 and 2016 in the Yunnan R. tanezumi plague focus and combined genomic analysis with epidemiological data to infer genomic diversity, spread, and transfer patterns of Y. pestis across epidemic-silent intervals.Continuous surveillance data of more than 60 years and density sampling of 16.In this study, 356 strains were isolated from 11 prefectures or cities in Yunnan Province, China, and Myanmar, near the China-Myanmar border, between 1952 and 2016, spanning two epidemic-silent cycles Fig. . Among tY. pestis in Yunnan during the two epidemic-silent cycles, we reconstructed the phylogeny of 356 isolates from the Yunnan R. tanezumi plague. Ninety-six percent of the strains (343 strains) were assigned to phylogroup 1.ORI2, which is associated with the third historic pandemic   was higher than those in Sublineage2  Fig. . TherefoY. pestis during the two epidemics in Yunnan, we integrated the information of phylogenetic relationships, sampling dates, and geographic distribution of the 343 Y. pestis strains. Phylogeographic analysis revealed that strains of Cycle1, mostly corresponding to Sublineage1, were distributed in western Yunnan, including Dehong Dai-Jingpo Autonomous Prefecture (DH), the Dali Bai Autonomous Prefecture (DL), and Baoshan City (B)  Figs. . Most stSublineage2, including >99.29% of Cycle2 strains, can be further subdivided into four sub-clades (1.ORI2.2.3.1–1.ORI2.2.3.4) Figs. . Strains20, isolated in 1962 (Saigon-Nhatraung-62-3), 1967 , 1986 , and 1988 (P-14709) were grouped together and formed a distinct clade  holds that an ancestral-descendant connection exists between the two cycles. Y. pestis is preserved in individual hosts and vectors, or in the environment, such as soil, during the silent period, and its offspring can cause outbreaks again under suitable ecological conditions25. The second possibility (Hypothesis2) is that the introduction of new Y. pestis strains from other populations lead to the resurgence of plague in a specific focus26. This can occur through various scenarios, such as the rise of a distinct local population, spillover from an adjacent plague focus, or the introduction of a strain from a distant plague focus.Epidemiological evidence indicates that plague in a natural focus is characterized by periodic alternations between epidemics and silence. Normally, plague outbreaks cause a population decline of local susceptible hosts so the population size falls below the threshold to maintain the epidemic, which then leads to the silence phase of plague in the focus. Following years or decades of recovery, the number of R. tanezumi plague provides an ideal sample set to test the above assumptions. The surveillance here indicated that between 1952 and 2016, it experienced two epidemic periods and two silent periods  and Cycle2 (including Epidemic2 and Silent2) could be subdivided into two sister clades: Sublineage1 (including 98.33% of Cycle1 strains) and Sublineage2 (including 99.29% of Cycle2 strains), respectively  DH strains collected in Epidemic1, no obvious pattern could be summarized for spread during this period.We also inferred the spread pattern and possible origin of 27. Epidemiological data also indicated that DH was the area where plague first emerged and last disappeared during Epidemic2, and Y. pestis strains could be isolated in this region throughout the epidemic. Given that DH is a prefecture situated on the western edge of Yunnan Province and bordering northern Myanmar  were attributed to two independent migration events from the Central Highlands occurring in the early 1980s, a decade before the first human cases were reported. They propose that Y. pestis might survive in local wildlife before transmission to humans. This could also be the case for the Yunnan plagues, as human plague appeared four years later than the epizootic plague in Epidemic216.Epidemic-silent cycles of plague have been reported in other countries, with the plague in Mahajanga, a northwestern coastal city in Madagascar, being well-studiedY. pestis strains in Southeast Asia, it is difficult to investigate the origin of Yunnan R. tanezumi plague epidemics. Second, although no significant genomic variations in the strains responsible for Cycle1 and Cycle2 were identified, a bacterial pathogenicity experiment is still needed to determine the difference between Y. pestis isolated from these two periods to further explore the mechanism of epidemic cycles. Finally, ecological and environmental dynamics in the niche, such as climate change, have been proven to be related to plague outbreaks32. A comprehensive analysis of eco-evolutionary dynamics should be conducted in the future to clarify the causes and mechanisms, leading to the resurgence of the plague epidemic.Our study has several limitations. First, a sampling bias should be noted. The severity of plague was comparable between DL and DH during the Epidemic1 period. However, because of staffing shortages and material and financial resources in the 1950s, Epidemic1 strains were rarely collected in DH. Thus, there is insufficient genomic evidence to infer the origin and spread pattern of Epidemic1. Together with insufficient sampling of Y. pestis strains sampled over 60 years from the Yunnan R. tanezumi plague focus, which were involved in two epidemic-silent cycles. We found that the plague in Yunnan Province originated from DH or its adjacent countries and spread from west to east. Importantly, we found that the two plague epidemics in Yunnan were caused by two sister clades that evolved in parallel from a common ancestor and showed different genetic diversity. Our results provide robust genomic evidence that the second epidemic was led by a common ancestor, rather than a descendant of Epidemic1. A similar pattern might be the cause of epidemic-silent cycles in other natural plague foci globally, which needs further verification using a suitable dataset in the future.In this study, we analyzed the genetic diversity and spread pattern of Y. pestis genomes were used in this study. Among them, 470 global isolates were downloaded from The National Center for Biotechnology Information GenBank on October 19, 2020  kit . Whole-genome sequencing was performed on the Illumina X-Ten platform with a 150-bp paired-end sequencing library. An average of 500 Mb of clean data were generated for each strain. The reads were then assembled using SPAdesY. pestis CO92 (1.ORI1 phylogroup strain) (accession no. NC_003143.1) using MUMmer (v3.0)34 to generate base alignments and identify SNPs. Sequencing reads were mapped to the reference to evaluate SNP accuracy for each strain using BWA (v0.7)36 and GATK (v3.8)38. Only high-quality SNPs, with base quality >20 , supported by >10 reads, and present in the core genome shared by at least 95% of the total strains, were retained for further analysis. SNPs located in repetitive regions were removed from the SNP dataset.All assemblies were compared to the reference genome, Y. pestis strains, which were concatenated to construct a maximum likelihood tree using IQ-TREE (v1.6)39 under the Generalized Time Reversible (GTR) model with 100 bootstrap replicates to assess tree topology support40. Phylogenic analysis showed that 96.35% (343/356) of the Yunnan Y. pestis strains belonged to phylogroup 1.ORI2.A total of 3851 SNPs were identified in the 826 Y. pestis strains, with CO92 as the outgroup.To obtain a reliable topology with a high resolution of 1.ORI2 strains, we recalled the SNPs for the 343 Yunnan strains within this clade based on the same pipeline. Finally, 263 high-quality and non-homoplastic SNPs were identified and imported into Applied Maths Bionumerics 6.6 software to build a minimum spanning (MS) tree with hypothetical intermediate nodes for all 343 41 was employed to estimate the temporal signal before utilizing the concatenated sequences of SNPs, sampling locations, and dates of strains for discrete phylogeographic analysis in the BEAST software (v1.10)40 to speculate the spread routes of Y. pestis in Yunnan. We selected the GTR+γ substitution model, uncorrelated relaxed clock for substitution rate, constant population size, and Bayesian skyline for tree priors to perform Bayesian analysis. For each analysis, three independent Markov Chain Monte Carlo chains of 107 cycles for sub-clade 1.ORI2.2.3.3, and 106 for sub-clade 1.ORI2.2.3.4 were carried out, respectively, and then combined using LogCombiner (v2.6). Tracer (v1.7)42 was used to access the convergence and ensure that the effective sample sizes of all relevant parameters were above 20043. TreeAnnotator (v1.10) was used to generate a maximum clade credibility (MCC) tree, with the first 10% of the states excluded as burn-in. The MCC tree was visualized and modified using FigTree (v1.4) (http://tree.bio.ed.ac.uk/software/figtree/).TempEst (v1.5)Further information on research design is available in the Supplementary InformationDescription of Additional Supplementary FilesSupplementary Data 1Supplementary Data 2Supplementary Data 3Reporting Summary


Moringa stenopetala  plant material is not studied yet. Thus, parts of the plant has been studied as bio adsorbents for removing toxic manganese ion from aqueous solutions in batch adsorption model. The maximum percent removal of manganese ion obtained from laboratory synthetic wastewater at equilibrium are 96.05 %, 98.90 % and 97.93 % by M. stenopetala plant leaf, bark and seed, respectively. However, the use of M. stenopetala plant leaf procedures an intensive color with unpleasant odor, which is inauspicious. Therefore, M. stenopental plant leaf was no longer examined for isotherm and kinetics studies. The fitness of adsorption data were confirmed based on the value of correlation coefficient (R2). Thus, adsorption by bark best fits of Temkin model with R2 value of 0.9707, while adsorption by seed follows the Langmuir model with R2 value of 0.9733. Adsorption kinetics result indicates that pseudo second-order model well fitted with R2 value of 0.9912 and 0.9947 for bark and seed adsorbents, respectively. Additionally, the applicability of laboratory-developed method was also evaluated on a multicomponent real sample taken from KK textile industry from Addis Abeba, Ethiopia. After characterization, the percentage removal of manganese ion were 79.53 % and 88.93 % for bark and seed, respectively. This achievement is promising and in a good agreement with the results of single component laboratory synthetic wastes.Removal of heavy metal ions from industrial effluents using environmental friendly bioadsorbents is currently promising approach. However, removal of manganese metal ion via   Industrial waste dispose into enviromental segments is a significant issue that affects the biota in the receiving environment. Heavy metal concentrations in industrial waste effluents from the metal-finishing and metallurgical sectors are frequently high and pose major environmental pollution issues. One of the biggest issues is heavy metal contamination into waterbody and soil. In addition, toxic metal compounds came into contact with earth's surface can also leach into underground water after rain and snowfall. Consequently, several harmful metals may be present in the earth's water as well ,2. The bM. stenopetala plant parts including bark and seed were reported that contains different functinal groups/phytochemicals like OH-, amines, carbonyls, sulphates, arromatics, and the like which is responsible for adsorpative removal of metal ions from industrial influents [Heavy metals and harmful compounds can be removed from industrial effluents using a variety of techniques, including chemical precipitation, ion exchange, coagulation/flocculation, reverse osmosis, and others . The majnfluents .M.stenopetala seed and bark as adsorbent material for manganese ion adsorption removal in batch adsorption model.Nowadays, there is a wide expansions of industries established for the manufacture of processed goods along with excessive wastes throughout Ethiopia. However industrial wastes are not successfully treated before disposed into the nearby environmental segments. The economy of the country is not strong enough to use more advanced waste treatment technology. As a result, there is an urgent need to develop low-cost, effective and biofriendly adsorbents that effectively remove wastes. In line with this, there are reports that natural adsorbents constitute an excellent alternative for chemical remediation of heavy metals from industrial effluents . Kebede M. stenopentala plant materials from wastewater. The results made by this study confirms excellent, enviromental friendly and inexpensive adsorbents for removal of manganese metal ion from industrial effluents.The main objective of this study was to investigate manganese ion removal efficiency of 2M.stenopetala plant samples  were collected from koso share village, near Arbaminch in Southern Ethiopia, which is 500 km from Addis Ababa. M.stenopetala tree leaf, bark and seed were washed by distilled water to remove any contaminates. Then, seed was dried for 24 h at 110 °C and bark was dried at 105 °C in an oven overnight [, [vernight . The drinight [, .2.14.H2O in double distilled water [Analytical-grade reagents and deionized water were used in this study. 1000 mg/L of a synthetic stock solution of manganese was prepared using MnSOed water . The wor2.2m.stenopetala plant parts characterization taken from Arbaminch, Ethiopia confirmed the availablity of phytochemicals with a wide ranges of organic functional groups as reported Kebede and his coworkers in 2018 [The  in 2018 ,11. Havi2.3The batch experiment was performed by adding desired amount of metal solution in 50 mL volumetric flask at desired adsorbent dose, pH, agitation speed and temperature. The solution was transforrmed in to 250 mL conical flask and shaken by mechanical shaker  at desired rpm for definite periods. Adsorbent dose, metal ion concentration, contact time, agitation speed, pH, and temperature were optimized by continuous variation approach (studying one parameter keeping the others constant) . The dif2.4Wastewater samples were collected in triplicates from local industries found in Addis Abeba, KK textile industry. Three waste water samples were colleted from the discharge point of the effluent using preclear and acidified plastic bottles with a 30 min interval and it was mixed together. The sample was then, kept in ice bag and transported to laboratory for analysis.2.5and coworkers (2020) [i and Ce being the initial and metal concentration in mg/L at equilibrium, respectively.The percent of metal removed, R, was calculated according to Mahmoud, A. E. s (2020)  given asqe (mg/g), was calculated using equation in 2021 [For isotherm and kinetic study the data were taken at the equilibrium conditions using the equations dislayed under  in 2021 ,20.2qe= are bark(98.90 %) > seed (97.93 %) > leaf (96.05 %).From the batch adsorption study: result indicated in 52 are regarded as a measure of the goodness-of-fit of experimental data. Also values of RL found to be between 0 and 1 indicating that both adsorbents are favorable for adsorption of Manganese ion from aqueous solution. Additionally, the adsorption intensity (n) is greater than unity implies that the forces within the surface layer are attractive confirming manganese ion is favorably adsorbed by M.stenopetala plant (bark and seed) [To find the most appropriate model, the data were fitted to different isotherm models and computed using equations presnted in nd seed) ,33. PartR2 value of the isotherms showed both isotherms fit well. In comparison with each other, the Langmuir model was able to adequately describe the adsorption of Manganese ion by seed with R2 value of 0.9733. While, Temkin model successfully describes the adsorption of Manganese ion by bark with R2 value of 0.9707, which reflects the occupation of the more energetic adsorption sites at first. It is highly probable that their adsorption sites are energetically non-equivalent. Similar results were also reported in previuos studies [M.stenopetala. This is obviously may be because of the surface morphology and functional groups available on each part of M.stenopetala plant.Where, 1/n), n = Freundlich Adsorption tendency,B = Related to the occupied surface of Temkin modelAs shown in  studies . The ads6To ascertain the adsorption kinetics of heavy metal ions, the kinetics parameters for the adsorption process were evaluated for various contact times by measuring the metal ion % adsorption. Following that, the data were regressed by different kinetics models, such as the Elovich kinetics equation, the Intraparticle diffusion kinetics equation, pseudo-first order kinetics equation and a pseudo-second order kinetics equation as presented in 2 value indicated in M. stenopetala bark and seed is regarded as pseudo-second order kinetics. Similar results were also found in Refs. [M.stenopetala.The value of correlation coefficient, in Refs. ,31. The 6.1The adsorbent materials have various types of phytochemicals with multifunctional groups as well as surface morphologies, according to previous study reports on the adsorption phenomena ,11,13,206.2M.stenopetala tree (bark and seed) as an adsorbent material was assessed by its application in treatment of industrial waste water sample. Textile industry waste water samples containing the selected metal ions were collected from local industries situated in Addis Abeba, KK textile industry. The physicochemical characteristics of industrial wastewater that could affetc adsorption process was studied as shown in The utilization of the 6.3M.stenopeta bark and seed as adsorbent materials for the removal of manganese ions from industrial effluents were evaluated as shown in Application of M. stenopetal plant bark remove about 79.53 % of manganese ion from real industrial wastewater taken from KK textile industry, in Addis Abeba, Ethiopia. In the other hand the seed adsorption is 88.93 % of manganese from same real wastewater taken from KK textile industry. According to the obtained results on adsorption removal of mananese ion from KK textile industry, the seed is more efficient in adsorption of manganese ion than the bark (seed- 88.93 % > bark −79.53 %). It shows a slight decrease of adsorption efficiency comparing to single manganese ion containing synthetic wastewater due to computation of other ions from real wastewater to occupy adsorption sites. This result indicates that M.stenopetala plant parts are a suitable cheap and effective adsorbent for the removal of manganse metal ions from industrial effluents.From this result, it is possible to suggest adsorbent effectiveness of manganese metal ion holding the optimum conditions obtained using laboratory prepared synthetic wastewater. M. stenopetala bark and seed were compared with literature values shown in M. stenopetala bark and seed adsorbents for remediation of environment from manganese pollution. The antagonistic effect of heavy metal removal under ideal circumstances was confirmed by the results in Adsorption efficiency of 7M.stenopetal plant bark and seed adsorbent materials in lab scale. The maximum adsorption efficiency of bark and seed using single component synthesic wastewater are 98.90 % and 97.93 % while in multicomponent real wastewater are 79.53 % and 88.93 respectively. Based on the obtained results, the researcher concluded that the bark and seed of M.stenopetala plant parts are effective adsorbents for remoal of manganese ion from industrial enffluents. This study found that adsorption parameters such as pH, contact time, adsorbent dosage, temperature, and stirring speed affect Manganese ion adsorption. The equilibrium data were applied to the Langmuir, Temkin and Freundlich isotherm models and pseudo first order, pseudo second order kinetics, Elovich and Intraparticle diffusion kinetics model. It was found that the adsorption of manganese by bark well fitted to the Temkin isotherm model while Langmuir isotherm successfully describes the adsorption of Manganese ion by seed very well. Among the kinetics models, the pseudo second order kinetic model best fits for the kinetics study that describes the chemosorption mechanisms of adsorption. The developed method is promising in treatment of industrial wastes. In conclusion, this study confirms a potential of environmental friendly M. stenopetala plant bark and seed as an adsorbent in removing manganese ion from wastewater. The results obtained are therefore very encouraging for developing industrial application of bio-adsorption technique.This study evaluated the removal of manganese ion from synthetic wastewater using a low-cost and effective No funding was received for conducting this study.The data used to support the findings of this study are included in this report.Ashenafi Zeleke Melaku: Writing – review & editing, Writing – original draft, Validation, Project administration, Methodology, Investigation, Funding acquisition, Formal analysis, Data curation, Conceptualization.There is no conflict of interest in this study.


Quantifying the biomechanical properties of the whole eye globe can provide a comprehensive understanding of the interactions among interconnected ocular components during dynamic physiological processes. By doing so, clinicians and researchers can gain valuable insights into the mechanisms underlying ocular diseases, such as glaucoma, and design interventions tailored to each patient’s unique needs.The aim of this study was to evaluate the feasibility and effectiveness of a multifocal acoustic radiation force (ARF) based reverberant optical coherence elastography (RevOCE) technique for quantifying shear wave speeds in different ocular components simultaneously.ex vivo. RevOCE measurements were conducted using an excitation pulse train consisting of 10 cycles at 3 kHz, followed by data processing to produce a volumetric map of the shear wave speed.We implemented a multifocal ARF technique to generate reverberant shear wave fields, which were then detected using phase-sensitive optical coherence tomography. A 3D-printed acoustic lens array was employed to manipulate a collimated ARF beam generated by an ultrasound transducer, producing multiple focused ARF beams on mouse eye globes The results show that the system can successfully generate reverberant shear wave fields in the eye globe, allowing for simultaneous estimation of shear wave speeds in various ocular components, including cornea, iris, lens, sclera, and retina. A comparative analysis revealed notable differences in wave speeds between different parts of the eye, for example, between the apical region of the cornea and the pupillary zone of the iris (The study demonstrated the effectiveness of RevOCE based on a non-invasive multifocal ARF for assessing the biomechanical properties of the whole eyeball. The findings indicate the potential to provide a comprehensive understanding of the mechanical behavior of the whole eye, which could lead to improved diagnosis and treatment of ocular diseases. For example, measuring the biomechanical properties of the cornea and sclera can help detect the early signs of glaucoma, a condition that can be characterized by increased intraocular pressure (IOP).,,Recent advances in innovative techniques for measuring the biomechanical properties of individual ocular tissues hold great promise for improving our understanding of ocular biomechanical properties. For example, the ocular response analyzer,in vivo application and hence may influence measurement results. Additionally, the resolution of USE is relatively low.Several studies have demonstrated promising outcomes in estimating the biomechanical properties of multiple ocular components simultaneously. One such study involved measuring displacement in response to eye globe inflation using multiple cameras and a laser displacement sensor to enable the assessment of regional variations in the stiffness of the cornea and sclera.,OCEIn this context, the aim of the current study is to demonstrate the capability of a non-invasive RevOCE in assessing the biomechanical properties of the entire eye globe with high mechanical resolution and contrast. We present a multifocal acoustic radiation force (ARF) system, which was implemented using an array of acoustic lenses and a single-element ultrasound transducer as an excitation source to induce reverberant shear wave field in mouse eyeballs.22.1Experiments were conducted on three freshly exercised mouse eyeballs (within 2 h after euthanasia). The eyeballs were cleaned of extraocular tissues, maintained in 1 X phosphate-buffered saline (PBS) solution, and frequently hydrated during imaging. The average IOP of the eyeballs, as measured by a rebound tonometer  before the start of OCE experiments, was 2.2As shown in During OCE imaging, the eyeball was mounted on a soft gelatin phantom  program. First, the phase shift, ,,,,en-face frames of each 3D particle velocity volume. Wave speed results in different regions of the eye were compared using Student’s t-test.The 4D OCE data  from the two-step measurement were processed to reconstruct a 3D map of the elastic wave speed in the whole eyeball using a custom MATLAB33.1The structural image of the whole eyeball was reconstructed from the M-C OCT scan. 3.2en-face images positioned in parallel planes orthogonal to the anterior-posterior pole. en-face OCT images (top row) taken from different en-face positions and instantaneous wave motion snapshots (bottom row) at The reverberant shear wave field induced in the eye globe can be visualized by taking the instantaneous particle velocity map in various 3.3Similarly, the wave speed map in the posterior segment shown in 3.4en-face speed maps displayed in The ability to quantify the biomechanical properties of the entire eye globe offers a significant advantage by allowing for the evaluation of the localized stiffness of specific ocular components such as the cornea. In 4,,,,,,,–The purpose of this study was to explore the potential of a novel RevOCE technique for quantifying the biomechanical properties of the whole eyeball. A multifocal ARF method was utilized to induce reverberant shear wave fields, which were then imaged using PhS-OCT. The results show that the technique can be used to quantify the biomechanical properties of the whole eyeball, providing insight into the relative differences in the stiffness of different ocular components of the intact whole eyeball. For the experimental conditions of the current study, the wave speed was found to be the greatest in the sclera, approaching ,,,,The technique also allowed for the evaluation of localized elastic wave speed within specific ocular components. For instance, the cornea showed regional variation in elastic wave speed, with higher elastic wave speed observed in the central regions and lower speed towards the peripheral regions. This finding is consistent with prior studies,The elastic wave speed profile in the posterior segment in proximity to the optic nerve revealed a gradual increase from the inferior (in vivo conditions. Future work will consider allowing excitation orientation on the same side of the objective lens to enable in vivo assessment, such as with an imaging hole as demonstrated with ring-shaped transducers.,While this study successfully determined the elastic wave speed of various ocular tissues using OCE, there are some limitations that could be addressed in future work. First, the current OCT system has a limited axial imaging range, which necessitates a two-step measurement process. To overcome this limitation, future work may utilize longer-range OCT systems like swept source OCT systems,,,in vivo applications. By providing a more complete understanding of the interrelationship of the biomechanical properties of the different ocular components, this technique can have important implications for both research and clinical practice. For instance, it can help in analyzing the effect of a particular disease on different ocular components, which can lead to more effective diagnosis and treatment strategies that may have been unavailable due to focusing on a specific ocular component only. Furthermore, the ability to concurrently evaluate multiple ocular components is especially useful in monitoring disease progression, where the effect reflected on secondary ocular component due to a diseased ocular component would carry significant value, such as improved diagnostic efficacy. It can also be used to assess the efficacy and side effects of treatment strategies, such as corneal cross-linking for keratoconus,,Generally, the RevOCE technique employed in this study offers an unprecedented opportunity to evaluate the biomechanical properties of various ocular components simultaneously. This technique stands out for its superior elasticity resolution when compared to the USE previously reported