Clinicopathological features of rhabdomyosarcoma with novel FET::TFCP2 and TIMP3::ALK fusion: report of two cases and literature review.

AIMS: The aim of this study was to evaluate the clinicopathological features, immunophenotype, differential diagnosis, molecular genetic features and prognosis of spindle cell rhabdomyosarcoma with TFCP2 rearrangement. METHODS: Two cases of spindle cell rhabdomyosarcoma with FET::TFCP2 gene fusion were included in this study. Samples were collected and evaluated through histological observation, immunohistochemistry, fluorescence in-situ hybridisation and high-throughput gene sequencing and previous findings. RESULTS: The tumour tissues mainly comprised spindle cells and epithelioid cells, which expressed striated muscle markers, and exhibited high expression levels of CK and ALK protein markers. Molecular detection showed that the FET::TFCP2 gene was fused. A rare case with TIMP3::ALK and FUS::TFCP2 double-fusion was observed in this study. CONCLUSIONS: A case with double fusion of ALK and TFCP2 was reported in rhabdomyosarcoma for the first time in this study, which provides information on the molecular characteristic of the tumour. Spindle cell rhabdomyosarcoma with FET::TFCP2 fusion is characterised by histological, immunohistochemical and genetic changes. The tumour is aggressive, with poor prognosis and poor response to radiotherapy and chemotherapy. The efficacy of targeted therapy for ALK should be explored through more clinical studies.

UnVELO: Unsupervised Vision-Enhanced LiDAR Odometry with Online Correction.

Due to the complementary characteristics of visual and LiDAR information, these two modalities have been fused to facilitate many vision tasks. However, current studies of learning-based odometries mainly focus on either the visual or LiDAR modality, leaving visual-LiDAR odometries (VLOs) under-explored. This work proposes a new method to implement an unsupervised VLO, which adopts a LiDAR-dominant scheme to fuse the two modalities. We, therefore, refer to it as unsupervised vision-enhanced LiDAR odometry (UnVELO). It converts 3D LiDAR points into a dense vertex map via spherical projection and generates a vertex color map by colorizing each vertex with visual information. Further, a point-to-plane distance-based geometric loss and a photometric-error-based visual loss are, respectively, placed on locally planar regions and cluttered regions. Last, but not least, we designed an online pose-correction module to refine the pose predicted by the trained UnVELO during test time. In contrast to the vision-dominant fusion scheme adopted in most previous VLOs, our LiDAR-dominant method adopts the dense representations for both modalities, which facilitates the visual-LiDAR fusion. Besides, our method uses the accurate LiDAR measurements instead of the predicted noisy dense depth maps, which significantly improves the robustness to illumination variations, as well as the efficiency of the online pose correction. The experiments on the KITTI and DSEC datasets showed that our method outperformed previous two-frame-based learning methods. It was also competitive with hybrid methods that integrate a global optimization on multiple or all frames.

Two novel prognostic models for ovarian cancer respectively based on ferroptosis and necroptosis.

BACKGROUND: Platinum-resistant cases account for 25% of ovarian cancer patients. Our aim was to construct two novel prognostic models based on gene expression data respectively from ferroptosis and necroptosis, for predicting the prognosis of advanced ovarian cancer patients with platinum treatment. METHODS: According to the different overall survivals, we screened differentially expressed genes (DEGs) from 85 ferroptosis-related and 159 necroptosis-related gene expression data in the GSE32062 cohort, to establish two ovarian cancer prognostic models based on calculating risk factors of DEGs, and log-rank test was used for statistical significance test of survival data. Subsequently, we validated the two models in the GSE26712 cohort and the GSE17260 cohort. In addition, we took gene enrichment and microenvironment analyses respectively using limma package and GSVA software to compare the differences between high- and low-risk ovarian cancer patients. RESULTS: We constructed two ovarian cancer prognostic models: a ferroptosis-related model based on eight-gene expression signature and a necroptosis-related model based on ten-gene expression signature. The two models performed well in the GSE26712 cohort, but the performance of necroptosis-related model was not well in the GSE17260 cohort. Gene enrichment and microenvironment analyses indicated that the main differences between high- and low- risk ovarian cancer patients occurred in the immune-related indexes, including the specific immune cells abundance and overall immune indexes. CONCLUSION: In this study, ovarian cancer prognostic models based on ferroptosis and necroptosis have been preliminarily validated in predicting prognosis of advanced patients treated with platinum drugs. And the risk score calculated by these two models reflected immune microenvironment. Future work is needed to find out other gene signatures and clinical characteristics to affect the accuracy and applicability of the two ovarian cancer prognostic models.

LncRNA-RMRP promotes nucleus pulposus cell proliferation through regulating miR-206 expression.

Long noncoding RNAs (LncRNAs) are involved in the pathogenesis of intervertebral disc degeneration (IDD). However, the biological function and expression of RMRP were still unclear. In our study, we showed that RMRP expression was up-regulated in degenerated NP tissues compared to normal NP samples, and higher RMRP expression was associated with the disc degeneration grade. Further studies indicated that ectopic expression of RMRP enhanced NP cell growth and also enhanced the expression of ki-67, PCNA and cyclin D1 in the NP cell. Moreover, overexpression of RMRP promoted the expression of Type II collagen and aggrecan and suppressed the expression of MMP13 and ADAMTS4. In addition, we found that the expression of miR-206 was down-regulated in degenerated NP tissues compared to normal NP samples, and lower miR-206 expression was correlated with the disc degeneration grade. Interestingly, we indicated that miR-206 expression in NP tissues was negatively correlated with the expression of RMRP. Ectopic expression of miR-206 suppressed NP cell proliferation and suppressed the expression of Type II collagen and aggrecan and enhanced the expression of MMP13 and ADAMTS4. Furthermore, we demonstrated that overexpression of RMRP increased NP cell growth and regulated ECM expression through targeting miR-206. These results suggested that lncRNA-RMRP promoted the progression of IDD through targeting miR-206, providing an attractive new therapeutic approach for the treatment of IDD disease.

miR-423-5p Inhibits Osteosarcoma Proliferation and Invasion Through Directly Targeting STMN1.

BACKGROUND/AIMS: Increasing evidences suggest that dysregulated expression of miRNAs contributes to the progression of various tumors. However, the underlying function of miR-423-5p in osteosarcoma remains unexplored. METHODS: The expression of miR-423-5p and STMN1 were determined in osteosarcoma samples and cell lines via quantitative real-time PCR. Colony formation and Cell Counting Kit-8 (CCK-8) assays were performed to measure cell proliferation ability and transwell analysis was used to detect cell invasion, and dual luciferase reporter assay was perform to analysis the interaction between the miR-423-5p and STMN1. RESULTS: The expression levels of miR-423-5p and STMN1 in the osteosarcoma tissues and cell lines were measured by qRT-PCR. Cell viability was determined using the clone formation and CCK-8 assays. A dual-luciferase reporter and Western blot were performed to stdudy the target gene of miR-423-5p. Here, we showed that miR-423-5p expression was downregulated in osteosarcoma tissues and cell lines. However, the expression of stathmin1 (STMN1) was downregulated in osteosarcoma tissues and cell lines. Moreover, STMN1 expression level was negatively correlated with the miR-423-5p expression in the osteosarcoma tissues. We identified STMN1 was a direct target gene of miR-423-5p in osteosarcoma cell. Overexpression of miR-423-5p inhibited osteosarcoma cell proliferation, colony formation and invasion. Furthermore, we demonstrated that STMN1 was involved in miR-423-5p-mediated cell behavior such as cell proliferation, colony formation and invasion in the osteosarcoma cell. CONCLUSION: Our present study indicated that miR-423-5p acted as a tumor suppressor gene in osteosarcoma partly through inhibiting STMN1 expression.

Amine-Functionalized Silica Nanoparticles with Drug and Gene Co-Delivery for Anti-Angiogenesis Therapy of Breast Cancer.

In our study, we report on the design and characterization of a combined angiogenesis therapy for breast cancer based on well-formed amine-functionalized silica nanoparticles (SLNs). The aminefunctionalized SLNs was employed to simultaneously deliver angiostatin (ANG) plasmid and candesartan (CD) to the same cancer cell. The well-formed ANG/CD/SLNs exhibited small particle size, reasonable positive charges, excellent loading of drug and gene in vitro. Moreover, aminefunctionalized SLNs were almost no cytotoxicity. ANG/CD/SLNs resulted in enhanced gene transfection compared to naked plasmid. More importantly, ANG/CD/SLNs as a co-delivery system achieved a stronger inhibitory effect on angiogenesis in vitro, possibly resulting from significant downregulation of vascular endothelial growth factor (VEGF) expression via different pathways. In particular, in vivo investigation on nude mice bearing MCF-7 xenografts confirmed that ANG/CD/SLNs codelivery system exerted strong anti-tumor efficacy by synergistic antiangiogenic mechanism.

Expression of Heat Shock Protein-27 (Hsp27) and P38MAPK in Esophageal Squamous Cell Carcinoma.

BACKGROUND Esophageal squamous cell carcinoma (ESCC) is a worldwide concern. This study looked at the relationship between the expression of differential proteins and the clinicopathological data and survival rate of ESCC patients to identify potential tumor markers for the growth and metastasis of ESCC. MATERIAL AND METHODS This study included 162 patients who underwent surgical excision for management of ESCC. Fresh ESCC tissue and adjacent normal tissue specimens were collected. Protein expressions were detected by western blotting. The expression of Hsp27 and P38MAPK were detected by immunohistochemistry in formalin-fixed paraffin embedded primary tissue specimens. RESULTS The rate of positive Hsp27 and P38MAPK expression in ESCC tissue were higher than in normal esophageal tissue (p<0.05). The expression of P38MAPK was related to the depth of infiltration (p<0.05). The expression of Hsp27 was correlated with lymph node metastasis (p<0.05), but not with age, depth of infiltration, or tumor size. ROC were plotted to estimate the significance of the diagnosis: for Hsp27, AUC=0.735 (p<0.05), for P38MAPK, AUC=0.882 (p<0.05). CONCLUSIONS The expression of Hsp27 and P38MAPK plays a role in ESCC development. Hsp27 and P38MAPK could be used as prognostic factors in ESCC.

A L0 sparse analysis prior for blind poissonian image deconvolution.

This paper proposes a new approach for blindly deconvolving images that are contaminated by Poisson noise. The proposed approach incorporates a new prior, that is the L0 sparse analysis prior, together with the total variation constraint into the maximum a posteriori (MAP) framework for deconvolution. A greedy analysis pursuit numerical scheme is exploited to solve the L0 regularized MAP problem. Experimental results show that our approach not only produces smooth results substantially suppressing artifacts and noise, but also preserves intensity changes sharply. Both quantitative and qualitative comparisons to the specialized state-of-the-art algorithms demonstrate its superiority.

Noncentral catadioptric camera calibration using a generalized unified model.

This Letter proposes a generalized unified model (GUM) for the calibration of noncentral catadioptric cameras. Releasing the constraint on the projection center and the orientation of the imaging plane that the traditional unified projection model has, the new model is able to well compensate the misalignment between the mirror and the camera. Being a compact and approximate central model, the GUM inherits the flexibility and simplicity from the unified model while maintaining accuracy even under severe misalignment. The calibration algorithm to compute the describing parameters of the model is also given. With the GUM, the calibration of central or noncentral systems could be treated with equal simplicity (or complexity). Experiments on both synthetic data and real images proved our success.

3D LIDAR-camera extrinsic calibration using an arbitrary trihedron.

This paper presents a novel way to address the extrinsic calibration problem for a system composed of a 3D LIDAR and a camera. The relative transformation between the two sensors is calibrated via a nonlinear least squares (NLS) problem, which is formulated in terms of the geometric constraints associated with a trihedral object. Precise initial estimates of NLS are obtained by dividing it into two sub-problems that are solved individually. With the precise initializations, the calibration parameters are further refined by iteratively optimizing the NLS problem. The algorithm is validated on both simulated and real data, as well as a 3D reconstruction application. Moreover, since the trihedral target used for calibration can be either orthogonal or not, it is very often present in structured environments, making the calibration convenient.

USP14 predicts poorer survival outcomes and promotes tumor progression in endometrial carcinoma by activating NF-κB signaling.

Ubiquitin-specific protease 14 (USP14), a member of the USP family, which catalyzes ubiquitin cleavage from a range of protein substrates, has been found dysregulated in several cancers. Our aim is to explore the functions and mechanism of USP14 in endometrial carcinoma (EC). Quantitative real-time PCR (qRT-PCR) and western blot (WB) were used to assess USP14 levels in EC tissues and adjacent nontumor tissues. USP14 overexpression or knockdown models were adopted to determine USP14-mediated effects on EC cell proliferation, migration, invasion, apoptosis, and epithelial-mesenchymal transition (EMT). The xenograft tumor experiment checked the effect of USP14 overexpression on tumor cell growth. Furthermore, the upstream signaling pathway of USP14 was predicted by bioinformatics. Consequently, EC tissues exhibited USP14 overexpression compared to normal paracancerous nontumor tissues. USP14 presence was linked to an adverse prognosis in EC cases. Functionally, USP14 overexpression reduced apoptosis and increased cell migration, invasion, and EMT in vivo and ex vivo. USP14 knockdown had the opposite effect. Mechanistically, NF-κB pathway activation occurred through the inhibitory effect of USP14 on I-κB expression. Conversely, NF-κB pathway inhibition attenuated USP14-mediated carcinogenic effects. Additionally, there existed a binding interaction between miR-124-3p and the 3'-UTR of USP14, resulting in USP14 activity inhibition. In summary, our research indicates that the involvement of miR-124-3p in USP14 regulation contributes to exacerbated EC progression through NF-κB pathway activation. The modulation of this pathway may be a new strategy for treating EC.

Offline-Online Associated Camera-Aware Proxies for Unsupervised Person Re-Identification.

Recently, unsupervised person re-identification (Re-ID) has received increasing research attention due to its potential for label-free applications. A promising way to address unsupervised Re-ID is clustering-based, which generates pseudo labels by clustering and uses the pseudo labels to train a Re-ID model iteratively. However, most clustering-based methods take each cluster as a pseudo identity class, neglecting the intra-cluster variance mainly caused by the change of cameras. To address this issue, we propose to split each single cluster into multiple proxies according to camera views. The camera-aware proxies explicitly capture local structures within clusters, by which the intra-ID variance and inter-ID similarity can be better tackled. Assisted with the camera-aware proxies, we design two proxy-level contrastive learning losses that are, respectively, based on offline and online association results. The offline association directly associates proxies according to the clustering and splitting results, while the online strategy dynamically associates proxies in terms of up-to-date features to reduce the noise caused by the delayed update of pseudo labels. The combination of two losses enables us to train a desirable Re-ID model. Extensive experiments on three person Re-ID datasets and one vehicle Re-ID dataset show that our proposed approach demonstrates competitive performance with state-of-the-art methods. Code will be available at: https://github.com/Terminator8758/O2CAP.

Emerging Roles of Long Non-Coding RNAs in Ankylosing Spondylitis.

Ankylosing spondylitis (AS) is a chronic systemic autoimmune disease characterized by inflammation, bone erosion, spur formation of the spine and the sacroiliac joints. However, the etiology and molecular pathogenesis of AS remain largely unclear. Recently, a growing number of studies showed that long non-coding RNAs (lncRNAs) played critical roles in the development and progression of autoimmune and orthopedic conditions, including AS. Studies demonstrated that a myriad of lncRNAs (e.g. H19, MEG3, LOC645166) pertinent to regulation of inflammatory signals were deregulated in AS. A number of lncRNAs might also serve as new biomarkers for the diagnosis and predicting the outcomes of AS. In this review, we summarize lncRNA profiling studies on AS and the functional roles and mechanism of key lncRNAs relevant to AS pathogenesis. We also discuss their potential values as biomarkers and druggable targets for this potentially disabling condition.

An update on the roles of circular RNAs in spinal cord injury.

Spinal cord injury (SCI) is a disabling condition for which therapeutic options are limited. Increasing number of microarray and next-generation sequencing studies have demonstrated that SCI coincides with altered expression of circular RNAs (circRNAs) in the damaged tissue. Emerging functional evidence further pinpointed specific differentially expressed circRNAs (e.g., circ-HIPK3, cicRNA.7079, circRNA_01477, circRNA-2960, and circ_0001723) for their effects on cellular processes relevant to SCI repair and regeneration, including neuronal apoptosis, astrocyte activation, and neuroinflammation, via sponging SCI-related microRNAs. Although circRNAs and their target microRNAs appear to be good candidates for therapeutic exploitation in SCI, further investigation into the efficient delivery of these regulatory molecules in a cell-type specific manner is a pre-requisite for translating these basic discoveries into clinical benefits.

GLUT5 increases fructose utilization in ovarian cancer.

Background: Fructose is one of the most common dietary carbohydrates in the whole world, and recent studies have found that fructose consumption is closely related to the oncogenesis and development of tumors, however, very few studies have focused on the fructose in ovarian cancer. GLUT5 (Glucose transporter type 5), as a specific fructose transporter in mammalian cells, has also been found highly expressed in many cancers. Methods: In this study, we investigated the abilities of proliferation, colony formation, and migration of ovarian cancer cells in fructose medium, and then silenced GLUT5 in ovarian cancer cells to explore the role GLUT5 in fructose metabolism in ovarian cancer. Results: The results showed that the ovarian cancer cells had similar abilities of proliferation and migration in fructose medium and glucose medium, but silencing GLUT5 could significantly inhibit these abilities in fructose medium. Meanwhile, we found that GLUT5 was higher expressed in ovarian cancer tissues, and its expression correlated significantly with tumor malignancy and poor survival of ovarian cancer patients. Furthermore, the results of animal experiments also demonstrated that intake too much fructose could prominently increase tumor volume, and silencing GLUT5 could significantly inhibit tumor proliferation. Conclusion: In conclusion, we demonstrate that ovarian cancer cells could utilize fructose for their growth, and restricting the fructose intake or targeting GLUT5 may be efficacious strategies for ovarian cancer therapy.

LncRNA HIF1A-AS2 promotes osteosarcoma progression by acting as a sponge of miR-129-5p.

Increasing studies have demonstrated that long noncoding RNAs (lncRNAs) play vital roles in tumor development and progression. However, the relationship between osteosarcoma and HIF1AAS2 remains unknown. The expression of HIF1AAS2 and miR-129-5p was detected in osteosarcoma cell lines and samples via qRT-PCR. Cell Counting Kit-8 (CCK-8) and invasion assays were performed to determine cell proliferation and invasion ability, and a dual luciferase reporter assay was performed to determine the interaction between HIF1AAS2 and miR-129-5p. We showed that the expression of HIF1A-AS2 was upregulated in the osteosarcoma samples compared with the expression in noncancerous samples. Moreover, patients with high HIF1A-AS2 expression had a shorter overall survival. Ectopic expression of HIF1A-AS2 enhanced osteosarcoma cell proliferation, cell cycle progression and invasion. We found that overexpression of miR-129-5p decreased the luciferase activity of wild-type (WT) HIF1A-AS2 but not mutant HIF1A-AS2. Ectopic expression of HIF1A-AS2 suppressed miR-129-5p expression in MG-63 cells. We demonstrated that miR-129-5p was downregulated in osteosarcoma and was negatively associated with HIF1A-AS2 expression. Furthermore, ectopic expression of miR-129-5p suppressed osteosarcoma cell proliferation, cell cycle progression and invasion. In addition, overexpression of HIF1A-AS2 promoted cell proliferation, cell cycle progression and invasion of osteosarcoma cells through the modulation of miR-129-5p. These results indicated that HIF1A-AS2 might be a potential therapeutic target for osteosarcoma.

Changes in inflammatory factors and prognosis of patients complicated with non-alcoholic fatty liver disease undergoing coronary artery bypass grafting.

This study was designed to assess the relationship between changes in peripheral inflammatory factors of patients complicated with non-alcoholic fatty liver disease (NAFLD) undergoing coronary artery bypass grafting (CABG) before and after operation, and their prognosis. A total of 68 patients with stable angina pectoris treated in Xinjiang Hospital who underwent CABG at some point between August of 2013 and August of 2015 were enrolled in the study, and divided into the NAFLD group (n=31) and the non-NAFLD group (n=37) according to the presence of the condition or its absence. Peripheral blood was drawn from the patients before and at 24 h and 1 month after the operation, and the expression levels of high-sensitivity C-reactive protein (hsCRP), soluble CD40 ligand (sCD40L), intercellular adhesion molecule-1 (ICAM-1) and matrix metalloproteinase-9 (MMP-9) were measured via standard enzyme-linked immunosorbent assay. Our results showed the expression levels of hsCRP and sCD40L of patients in both groups reached a peak 24 h after operation; and there were statistically significant changes compared with the levels before the operation and at 1 month after operation (p<0.01). However, there were no statistically significant differences in the expression levels between the two groups (p>0.05). The expression levels of ICAM-1 at each time-point in both groups were increased after the operation, but the changes were not statistically significant (p>0.05). The expression levels of MMP-9 increased after the operation, and the levels at 1 month after operation were significantly higher than those before operation and at 24 h after the operation (p<0.01). Importantly, the expression levels of MMP-9 of patients in the NAFLD group at 1 month after operation were significantly higher than those of patients in the non-NAFLD group at the same time, and the differences were statistically significant (p<0.01). Finally, logistic regression analysis showed that the expression level of MMP-9 was an important influencing factor for cardiovascular events after CABG (OR=1.182, p<0.05). Based on our findings, the expression levels of inflammatory factors in peripheral blood in patients complicated with NAFLD undergoing CABG differ from those who are not complicated, and the MMP-9 levels may be closely related to the prognosis of patients complicated with NAFLD.

MicroRNA-103 Promotes Proliferation and Inhibits Apoptosis in Spinal Osteosarcoma Cells by Targeting p57.

Osteosarcoma is one of the most aggressive malignancies with poor prognosis rates. Many studies have demonstrated that miRNAs were involved in osteosarcoma, but the role of miR-103a in osteosarcoma remains elusive. In this study, we detected the expression levels of miR-103 in osteosarcoma and non-osteosarcoma tissues and cell lines. The binding effect of miR-103 on p57 was detected by luciferase reporter assay. After altering expressions of miR-103 or p57, viability, migration, invasion, and apoptosis of MG63 cells and expressions of proteins related with the JNK/STAT and mTOR pathways were all detected. We found the higher expression of miR-103 in osteosarcoma tissues and cell lines compared with non-osteosarcoma tissues and cell lines. miR-103 overexpression promoted survival, migration, and invasion of MG63 cells. Knockdown of miR-103a inhibited cell survival, migration, and invasion by upregulating the expression of p57, which was a target of miR-103. Moreover, miR-103a overexpression activated the JNK/STAT and mTOR pathways probably through inhibiting p57 expression. In conclusion, miR-103a acted as an oncogene in osteosarcoma, probably through activating the JNK/STAT and mTOR pathways by inhibiting p57 expression.

Bipolar Sealer Device Reduces Blood Loss and Transfusion Requirements in Posterior Spinal Fusion for Degenerative Lumbar Scoliosis: A Randomized Control Trial.

STUDY DESIGN: A prospective, randomized, controlled clinical study. OBJECTIVE: To determine the efficacy of bipolar sealer device in reducing perioperative blood loss and transfusions in degenerative lumbar scoliosis patients undergoing primary posterior spinal fusion. BACKGROUND: It has recently been used successfully in pediatric spine surgery, particularly in idiopathic and neuromuscular deformities. However, there is a dearth of literature on prospective study of the efficacy of bipolar sealer device in reducing perioperative blood loss and transfusions in patients undergoing degenerative lumbar scoliosis surgery. METHODS: A total of 100 consecutive degenerative lumbar scoliosis patients who had undergone primary decompression and posterior spinal fusion with segmental spinal instrumentation between June 2010 and June 2012 were prospectively randomized into 1 of 2 groups according to whether bipolar sealer device for intraoperative/postoperative blood management was used or not. Demographic distribution, perioperative blood loss, blood transfusion rate, the length of stay, and postoperative complications were analyzed. RESULTS: The operation time was significantly shorter in the study group than in the control group, 223.4 versus 248.9 minutes (P=0.026). There was significantly lower intraoperative estimated blood loss in the bipolar sealer group, 407 versus 696 mL (P=0.000). Of the patients with the use of bipolar sealer device, the mean red blood cell transfusion requirement during hospitalization was significantly less than the control group, 0.4 versus 1.1 U/patient (P=0.003). Furthermore, significant difference existed in allogenic blood transfusion rate between the 2 cohorts. Within the study group (with the use of bipolar sealer device), the entire perioperative allogenic blood transfusion rate was 18.0% compared with 40.0% of the control group (P=0.015). There were no complications related directly to the use of the bipolar sealer. CONCLUSIONS: Utilization of a bipolar sealer during correction of lumbar degenerative scoliosis may offer comparable hemostatic effects, without prohibitive cost or adverse drug-related risks.