Cellular Communication Networks Research Articles

Construction of a Localized and Long-Acting CCN2 Delivery System on Percutaneous Ti Implant Surfaces for Enhanced Soft-Tissue Integration

Soft-tissue integration (STI) plays an essential role in the long-term success of percutaneous Ti implants since it acts as a biological barrier that protects the soft and hard tissue around implants. Surface modification of Ti implants with drug-release properties to achieve soft-tissue regeneration has been proven to be effective in STI. However, the short-acting effect caused by the uncontrolled drug release of the topical delivery system limits long-term STI enhancement. Herein, a long-acting protein delivery system for Ti implants that involved micro-arc oxidation of Ti surfaces (MAO-Ti) and localized immobilization of cellular communication network factor 2 (CCN2) bearing mesoporous silica nanoparticles (MSNs) on MAO-Ti was prepared, namely, CCN2@MSNs-Ti. The CCN2 release study of CCN2@MSNs-Ti demonstrated a sustained-release profile for 21 days, which was able to maintain long-term stable STI. In addition, in vitro cell behavior evaluation results indicated that CCN2@MSNs-Ti could promote the STI-related biological response of human dermal fibroblasts via the FAK-MAPK pathway. More importantly, the system could effectively enhance STI after 4 weeks and proinflammatory factors in the soft tissue decreased significantly in a rat model of implantation. These results denote that CCN2@MSNs-Ti showed an appealing application prospect for enhanced STI around transcutaneous Ti implants, which would ultimately result in an increased success rate of percutaneous Ti implants.

Abstract 587: The Matricellular Protein Ccn1 Induces Aortic Aneurysms In Mice

Aortic aneurysm ( AA ) is a dilation of the aorta due to an arterial wall weakening. Chronic inflammation and oxidative stress have been associated with dysregulated vascular smooth muscle cell (VSMC) homeostasis and functional phenotype leading to loss of VSMCs, matrix remodeling, and AA formation. The matricellular protein CCN1 (cellular communication network factor 1) has been identified in a subpopulation of VSMCs at the early stage of phenotypic switching. To assess the involvement of CCN1 in AA, CCN1 expression was examined in the CaCl 2 -induced mouse AA model using Ccn1 lacZ/+ -knockin mice. CCN1 expression revealed by whole-mount aortic X-gal staining was intensely induced 3 days after the peri-aortic application of CaCl 2 and sustained more refined to the lesion areas. CCN1 was largely expressed in VSMCs and infiltrating macrophages. CCN1 binds to its receptor integrin α6β1 and promotes atherogenesis through prolonged elevations of inflammation, oxidative stress, and matrix remodeling, the similar driving forces underlying AA progression. To test the role of CCN1 in AA development, Ccn1 dm/dm mice, carrying the α6β1-binding-defective mutant allele Ccn1-dm , were tested in the mouse AA model. Ccn1 dm/dm mice exhibited remarkable resistance against CaCl 2 -induced AA comparing with wild-type control mice 4 weeks after treatment. The Ccn1 dm/dm aortic wall possessed well-structured elastic lamellae and organized VSMC layers with less infiltrating macrophages, inflammation (interleukin-1β and tumor necrosis factor-α), and oxidative stress (8-OHdG). MMP9 levels detected by immunostaining were induced by CaCl 2 in the wild-type aortic wall, but not detected in Ccn1 dm/dm mice. Together, our results demonstrate a critical role of CCN1 in AA formation through binding its receptor α6β1 to increase VSMC inflammation and oxidative stress. CCN1 can be used as a novel therapeutic target for AA.

Abstract 532: Deficiency of CCN3 in Smooth Muscle Cells Perturbs Vascular Homeostasis and Exacerbates Thoracic Aortic Dissection

Introduction: Thoracic aortic dissection (TAD) is a life-threatening aortic disease with a high mortality rate in the event of rupture. Key mechanisms underlying the development of TAD include extracellular matrix (ECM) degradation and vascular smooth muscle cell (SMC) reprograming. Cellular communication network factor 3 (CCN3), a matricellular protein highly expressed in SMC, exerts a myriad of context-dependent biological functions. However, its role in SMC homeostasis and in the development of TAD remains largely undefined. Methods: A single-cell sequencing (Sc-seq) dataset (GSE213740) from human aortic dissection patients was analyzed to determine the association between CCN3 expression and TAD. In newly weaned male CCN3-floxed mice harboring Myh11-driven inducible Cre, 4 consecutive tamoxifen injections induced CCN3 deletion in SMCs. Mice without Cre expression were subjected to the same procedures and were used as controls. After 1 week of washout, 4-week-old mice were given 0.5% BAPN (β-aminopropionitrile monofumarate) in their drinking water to induce TAD. Histologic and protein measurements were performed on TAD tissues from control and CCN3-SMC-KO mice. Sc-seq analysis was performed to recapitulate cell heterogeneities and transcriptome profile changes. The regulatory role of SMC-derived CCN3 in TGFβ signaling was evaluated by western blot and colocalization analyses. Results: Analysis of the human Sc-seq dataset revealed downregulation of CCN3 in aortas from patients with TAD. CCN3-positive SMCs have elevated ECM and elastin gene expression in comparison to CCN3-null SMCs. In vivo, concordant with increased mortality in CCN3-SMC-KO animals, SMC-specific deletion of CCN3 promotes BAPN-induced elastin fibers fragmentation and degradation. In vitro, SMC deficiency of CCN3 diminished TGFβ-induced elastogenesis. CCN3 binds to the TGFβ receptor on SMCs and augments TGFβ receptor phosphorylation, thereby activating Smads, upregulating SMC elastogenesis, and as a consequence, alleviates elastin fiber fragmentation and TAD development. Conclusion: CCN3 is a key determinate of SMC reprogramming and sustains vascular homeostasis. Our findings suggest a novel target for the prevention and treatment of TAD.

Effects of CCN6 overexpression on the cell motility and activation of p38/bone morphogenetic protein signaling pathways in pancreatic cancer cells

Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive cancer types that is highly resistant to conventional treatments, such as chemotherapy and radiotherapy. As the demand for more effective therapeutics for PDAC treatment increases, various approaches have been studied to develop novel targets. The cellular communication network (CCN) family is a matricellular protein that modulates various cellular functions, including cell adhesion, proliferation, migration, and invasiveness. Despite this, little is known about the role of CCN6 in PDAC. The current study investigated the role of CCN6 in PDAC by generating CCN6-overexpressing PANC-1 cells (PANC-1-CCN6) by infecting lentivirus particles containing CCN6. PANC-1-CCN6 induces cell viability and tumorigenesis than PANC-1 cells with empty vector (control). The PANC-1-CCN6 formed more colonies, and the size of spheroids increased compared to the control. The upregulation of CCN6 enhances the expression of bone morphogenetic proteins (BMPs) genes and the upregulation of p38 mitogen-activated protein kinases (MAPKs). In PANC-1-CCN6 cells, the levels of N-cadherin, VEGF, and Snail expression were higher than the control, while E-cadherin expression was lower, which is associated with upregulation of epithelial-to-mesenchymal transition (EMT). Consistent with the changes in EMT-related proteins in PANC-1-CCN6, the migratory ability and invasiveness were enhanced in PANC-1-CCN6. Xenografted PANC-1-CCN6 in immunocompromised mice exhibited accelerated tumor growth than the control group. In immunohistochemistry (IHC), the PANC-1-CCN6 xenografted tumor showed an increased positive area of PCNA and Ki-67 than the control. These results suggest that CCN6 plays a tumorigenic role and induces the metastatic potential by the p38 MAPK and BMPs signaling pathways. Although the role of CCN6 has been introduced as an antitumor factor, there was evidence of CCN6 acting to cause tumorigenesis and invasion in PANC-1.

P150 The Triptryium wilfordii derivative celastrol has anti-fibrotic effects in systemic sclerosis

Abstract Background/Aims Scleroderma (systemic sclerosis, SSc) is an autoimmune inflammatory-fibrosis syndrome that damages the skin and internal organs and is considered a prototypic complex fibrotic disease. In SSc, persistently activated myofibroblasts are maintained by an excessive, autocrine mechanotranductive/pro-adhesive signaling loop. Drugs targeting this pathway are therefore of likely therapeutic benefit in SSc. The mechanosensitive transcriptional co-activator, yes activated protein-1 (YAP1), is activated in SSc fibroblasts. The terpenoid plant-derivative celastrol has recently been identified as a YAP inhibitor; however, if celastrol can alleviate SSc fibrosis, and its underlying mechanism of action, is as yet unclear. Methods We cultured age-, sex-, and site-matched human dermal fibroblasts sampled from healthy individuals and patients with early onset (<18 month after first diagnosis) diffuse cutaneous scleroderma (systemic sclerosis, dcSSc) and treated with or without transforming growth factor β1 (TGFβ1, 4ng/ml) in the presence or absence of celastrol (500nM). We also subjected C57BL6J mice to the inflammatory-driven bleomycin-induced model of skin SSc, in the presence or absence of celastrol (28days, every day, 0.1U bleomycin, 1mg/kg celastrol). RNA expression was assessed using RNAseq and real-time polymerase chain reaction analysis. Protein expression was determined using Western blot and enzyme-linked immunosorbent assay. Fibrosis was monitored by hematoxylin and eosin and trichrome staining of tissue sections, and by using indirect immunofluorescence analysis with anti-alpha-smooth muscle actin (SMA) antibodies to detect myofibroblasts. Results In dermal fibroblasts, celastrol impaired the ability of TGFβ1 to induce an SSc-like pattern of gene expression, including the induction of cellular communication network factor 2 (CCN2), collagen I and TGFβ1 protein (N = 6, all p < 0.01). Celastrol also alleviated the persistent fibrotic phenotype of dermal fibroblasts cultured from lesions of SSc patients, including the overproduction of CCN2 and collagen (N = 3, p < 0.05). Finally, in the bleomycin-induced model of SSc dermatopathology, celastrol inhibited skin thickness/collagen deposition in vivo and blocked nuclear localization of YAP in SMA-positive myofibroblasts (all N = 6, p < 0.01). Conclusion Our data are consistent with the hypothesis that compounds, such as celastrol, that antagonize the YAP pathway warrant further consideration as potential treatments for SSc skin fibrosis. Disclosure A. Leask: Grants/research support; CIHR, NSERC, Arthritis Society of Canada. R.J. Stratton: None. S. Xu: None. P. Chitturi: None.

Revealing the roles of glycosphingolipid metabolism pathway in the development of keloid: a conjoint analysis of single-cell and machine learning.

Keloid is a pathological scar formed by abnormal wound healing, characterized by the persistence of local inflammation and excessive collagen deposition, where the intensity of inflammation is positively correlated with the size of the scar formation. The pathophysiological mechanisms underlying keloid formation are unclear, and keloid remains a therapeutic challenge in clinical practice. This study is the first to investigate the role of glycosphingolipid (GSL) metabolism pathway in the development of keloid. Single cell sequencing and microarray data were applied to systematically analyze and screen the glycosphingolipid metabolism related genes using differential gene analysis and machine learning algorithms (random forest and support vector machine), and a set of genes, including ARSA,GBA2,SUMF2,GLTP,GALC and HEXB, were finally identified, for which keloid diagnostic model was constructed and immune infiltration profiles were analyzed, demonstrating that this set of genes could serve as a new therapeutic target for keloid. Further unsupervised clustering was performed by using expression profiles of glycosphingolipid metabolism genes to discover keloid subgroups, immune cells, inflammatory factor differences and the main pathways of enrichment between different subgroups were calculated. The single-cell resolution transcriptome landscape concentrated on fibroblasts. By calculating the activity of the GSL metabolism pathway for each fibroblast, we investigated the activity changes of GSL metabolism pathway in fibroblasts using pseudotime trajectory analysis and found that the increased activity of the GSL metabolism pathway was associated with fibroblast differentiation. Subsequent analysis of the cellular communication network revealed the existence of a fibroblast-centered communication regulatory network in keloids and that the activity of the GSL metabolism pathway in fibroblasts has an impact on cellular communication. This contributes to the further understanding of the pathogenesis of keloids. Overall, we provide new insights into the pathophysiological mechanisms of keloids, and our results may provide new ideas for the diagnosis and treatment of keloids.

CCN1 expression is regulated by mechanical stimuli in tendons

Tendon overuse injuries are common, but the processes that govern tendon response to mechanical load are not fully understood. A series of experiments of in vitro and in vivo experiments was devised to study to the relationship between mechanical stimuli and the matricellular protein Cellular Communication Network Factor 1 (CCN1) in tenocytes and tendons. First, human and murine tenocytes were subjected to cyclic uniaxial loading in order to evaluate changes in CCN1 gene expression as a response to mechanical stimuli. Then, baseline Ccn1 gene expression in different murine tendons (Achilles, patellar, forearm, and tail) was examined. Finally, changes in Ccn1 expression after in vivo unloading experiments were examined. It was found that CCN1 expression significantly increased in both human and murine tenocytes at 5 and 10% cyclical uniaxial strain, while 2.5% strain did not have any effect on CCN1 expression. At baseline, the Achilles, patellar, and forearm tendons had higher expression levels of Ccn1 as compared to tail tendons. Twenty-four hours of immobilization of the hind-limb resulted in a significant decrease in Ccn1 expression in both the Achilles and patellar tendons. In summary, CCN1 expression is up-regulated in tenocytes subjected to mechanical load and down-regulated by loss of mechanical load in tendons. These results show that CCN1 expression in tendons is at least partially regulated by mechanical stimuli.

Spatial heterogeneity and Immune infiltration of cellular lysosomal pathways reveals a new blueprint for tumor heterogeneity in esophageal cancer.

Esophageal squamous cell carcinoma (ESCC) is a common Malignant tumor of digestive tract which have a potential association with lysosomal pathway. The purpose of this study was to explore the correlation between lysosome pathway and immune infiltration of ESCC. The cell type annotation of ESCC patients and the distribution of their gene markers were analyzed by single cell data. They were also grouped according to the expression of lysosomal pathways. Gene set variation analysis (GSVA) enriched pathway scoring, Cellchat cell communication was performed to demonstrate the tumour-associated pathway scores and interactions of different cell populations. Relevant differential genes were screened, prognostic risk markers were constructed and direct associations of lysosomal pathway-related gene risk scores with immune infiltration and tumour treatment drug sensitivity were assessed by algorithms. In cellular experiments, qPCR and flow cytometry were used to assess the role of the lysosomal pathway gene-MT1X on tumour cell development. ESCC single cell data were annotated into 7 Cluster clusters by t-sne downscaling analysis. Cellchat analysis revealed that the "MIF" cellular communication network is the main communication mode of the lysosomal pathway in ESCC cells. The lysosomal pathway genetic risk model was found to be significantly different from ESCC prognosis in both the training and validation groups. The lysosome pathway gene risk model was associated with treatment resistance in ESCC patients using oncopredict R package. The correlation between the expression of lysosomal-DEG and tumour immune infiltration and immune cell types by the MCPcounter method. Cellular assays showed that the lysosomal pathway gene MT1X was less expressed in oesophageal cancer cells than in normal oesophageal epithelial cells. Knockdown of MT1X significantly promoted the growth rate of oesophageal cancer cells. Based on the single cell sequencing technology and transcriptomic analysis, we confirmed that there is a close association between the lysosomal pathway and the immune infiltration and treatment sensitivity of ESCC, which may be a potential target for a new direction of ESCC therapy.

SPONDİLOEPİ (META) FİZYAL DİSPLAZİ ÖN TANILI BEŞ TÜRK ERKEK HASTADAKİ GENETİK ETYOLOJİNİN ARAŞTIRILMASI

OBJECTIVE: Skeletal dysplasias is a complex disease group characterized by disproportionate short stature and various orthopedic complications. X-Linked Spondyloepiphyseal Dysplasia Tarda is an X-linked inherited skeletal dysplasia accompanied by progressive spondyloepi(meta)physeal dysplasia and premature osteoartritis. The gene related to the disorder is trafficking protein particle complex 2 (TRAPPC2). Progressive pseudorheumatoid dysplasia (PPRD) is characterized by enlargement of the elbow joints and arthritis-like findings. It is an autosomal recessive subtype of skeletal dysplasia caused by mutations in cellular communication network factor 6 (CCN6) gene located on chrosomal region 6q21. In this study, it was aimed to diagnose five male individuals with an X-linked or autosomal recessive inheritance pattern, who have not been diagnosed for years, to identify possible treatments and to offer prenatal pmreimplantation genetic testing. MATERIAL AND METHODS: Five male siblings with skeletal dysplasia with an uncertain inheritance either X-linked or autosomal recessive pattern were included in this study, Whole Exome Sequencing (WES) was applied to the four affected cases. Sanger Sequencing was performed in one affected case and four healthy individuals. RESULTS: Homozygous c.210C>A (p.Cys70Ter) and homozygous c.302G>A (p.Gly101Glu) mutations in the CCN6 gene were found in all affected siblings. Thus, the final diagnosis after WES was autosomal recessive PPRD despite the possibility of an X-linked recessive pattern. CONCLUSIONS: This study presents a series of the oldest patients diagnosed with Progressive pseudorheumatoid dysplasia, normally a childhood disease, with an average age of 54.6. The p.Cys70Ter alteration is the most frequent pathogenic variant in Turkish patients. This study is also important in terms of showing that Progressive pseudorheumatoid dysplasia has no significant effect on life expectancy. At the same time, this study shows the progression of this disease and clinical findings that may accompany lifetime.

Abstract 3949: CYR61, a member of the CCN protein family, regulates IGF-1 in metastatic prostate cancer

Abstract The cysteine-rich angiogenic inducer 61 (CYR61) is a member of the cellular communication network (CCN) protein family that can be induced by growth factors. CYR61-specific functions are dependent on cellular context. In prostate cancer (PCa), CYR61 contributes to cell growth and survival through its interactions with extracellular ligands, such as integrins avb3 and a6b4, in the extracellular matrix space. Interestingly, CYR61 contains an insulin-like growth factor-binding protein domain suggesting that it may interact with insulin-like growth factor-I (IGF-1). High serum levels of IGF-1 are directly linked to PCa development and recently have been studied as a predictor of metastatic disease. Given the important roles that IGF-1 and CYR61 play in PCa and their potential interaction, it is critical to investigate their molecular interplay. This study is designated to specifically determine and characterize the molecular interaction between CYR61 and IGF-1 in metastatic PCa and to determine if this interaction contributes to aggressive tumor properties (e.g., proliferation, tumorsphere formation). Using immunoblotting, we demonstrated that CYR61 is upregulated in the metastatic cell line PC3 and downregulated in the docetaxel-resistant cell line PC3-DR. Furthermore, optimized knockdown of CYR61 using siRNA significantly reduced PC3 cell proliferation, viability, and prostasphere formation. To examine the underlying mechanisms associated with IGF-1 signaling, we assessed the activation of either the PI3/Akt and MAPK pathways in PC3 cells with CYR61 silencing. CYR61 siRNA-mediated knockdown decreased activation of the PI3/Akt pathway but did not affect the MAPK pathway. We also initiated studies to determine the co-localization of CYR61 and IGF-1 using immunofluorescence microscopy (IF). Based on the initial IF observations, we will proceed to examine if their interaction is primarily intra- or extra-cellular. For intracellular interactions, we will co-immunoprecipitate CYR61 with IGF-1 and for extracellular interactions, we will use the AVidity-based EXtracellular Interaction Screen (AVEXIS) system. Our goal is to establish the contribution of CYR61-IGF1 protein-protein interaction to PCa cell proliferation. Specifically, the correlation between PCa aggressiveness and circulating levels of CYR61 and IGF-1 is poorly studied. Their combined role may explain a proportion of aggressive PCa. Citation Format: Greisha L. Ortiz-Hernandez, Mya Walker, Leanne Woods-Burnham, Rick A. Kittles, Carlos A. Casiano, Susan L. Neuhausen. CYR61, a member of the CCN protein family, regulates IGF-1 in metastatic prostate cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3949.

An Efficient Mobile Gateway Selection and Discovery Based-Routing Protocol in Heterogeneous LTE-VANET Networks

Coupling cellular communication networks with vehicular ad hoc networks (VANET) can be a very interesting way out for providing Internet access to vehicles in the road. However, due to the several specific characteristics of VANETs, making an efficient multi-hop routing from vehicular sources to the Internet gateways through Long Term Evolution (LTE) technology is still challenging. In this paper, an Internet mobile gateway selection scheme is proposed to elect more potential vehicles to behave as gateways to Internet in VANETs. Therefore, the discovery and the selection of route to those mobiles gateways is carried out via an efficient multiple metrics-based relay selection mechanism. The objective is to select the more reliable route to the mobile gateways, by reducing the communication overhead and performing seamless handover. The proposed protocol is compared with one recent protocol based on packet delivery ratio, average end-to-end delay and overhead. The results show that the proposed protocol ameliorates significantly the network performance in the contrast of the other protocol.

BMP-1-induced GBA1 nuclear accumulation provokes CCN2 mRNA expression via importin-β-mediated nucleocytoplasmic pathway.

Bone morphogenetic protein (BMP)-1 is expressed by odontoblasts in the dentin-pulp complex. Although the functional effects of BMP-1 on the maturation of various preforms of proteins and enzymes involved in initiating mineralization have been widely observed, how BMP-1 affects cellular molecules remains unknown. We performed a comprehensive analysis of BMP-1-altered glycome profiles and subsequent assays to identify the target glycoproteins in human dental pulp cells (hDPCs) by a glycomic approach. In the presence of BMP-1, a lectin microarray analysis and lectin-probed blotting showed that α2,6-sialylation was significantly attenuated in insoluble fractions from hDPCs. Six proteins were identified by a mass spectrometry analysis of α2,6-sialylated glycoproteins purified using a lectin column. Among them, glucosylceramidase (GBA1) was found to accumulate in the nuclei of hDPCs in the presence of BMP-1. Moreover, BMP-1-induced cellular communication network factor (CCN) 2 expression, which is well known as the osteogenesis/chondrogenesis marker, was significantly suppressed in the cells transfected with GBA1 siRNA. Furthermore, importazole, a potent inhibitor of importin-β-mediated nuclear import significantly suppressed BMP-1-induced GBA1 nuclear accumulation and BMP-1-induced CCN2 mRNA expression, respectively. Thus, BMP-1 facilitates the accumulation of GBA1 in the nucleus through the reduction of α2,6-sialic acid, which potentially contributes to the transcriptional regulation of the CCN2 gene via importin-β-mediated nuclear import pathway in hDPCs. Our results offer new insights into the role of the BMP-1-GBA1-CCN2 axis in the development, tissue remodeling, and pathology of dental/craniofacial diseases.

Use of machine learning-based integration to develop a monocyte differentiation-related signature for improving prognosis in patients with sepsis

BackgroundAlthough significant advances have been made in intensive care medicine and antibacterial treatment, sepsis is still a common disease with high mortality. The condition of sepsis patients changes rapidly, and each hour of delay in the administration of appropriate antibiotic treatment can lead to a 4–7% increase in fatality. Therefore, early diagnosis and intervention may help improve the prognosis of patients with sepsis.MethodsWe obtained single-cell sequencing data from 12 patients. This included 14,622 cells from four patients with bacterial infectious sepsis and eight patients with sepsis admitted to the ICU for other various reasons. Monocyte differentiation trajectories were analyzed using the “monocle” software, and differentiation-related genes were identified. Based on the expression of differentiation-related genes, 99 machine-learning combinations of prognostic signatures were obtained, and risk scores were calculated for all patients. The “scissor” software was used to associate high-risk and low-risk patients with individual cells. The “cellchat” software was used to demonstrate the regulatory relationships between high-risk and low-risk cells in a cellular communication network. The diagnostic value and prognostic predictive value of Enah/Vasp-like (EVL) were determined. Clinical validation of the results was performed with 40 samples. The “CBNplot” software based on Bayesian network inference was used to construct EVL regulatory networks.ResultsWe systematically analyzed three cell states during monocyte differentiation. The differential analysis identified 166 monocyte differentiation-related genes. Among the 99 machine-learning combinations of prognostic signatures constructed, the Lasso + CoxBoost signature with 17 genes showed the best prognostic prediction performance. The highest percentage of high-risk cells was found in state one. Cell communication analysis demonstrated regulatory networks between high-risk and low-risk cell subpopulations and other immune cells. We then determined the diagnostic and prognostic value of EVL stabilization in multiple external datasets. Experiments with clinical samples demonstrated the accuracy of this analysis. Finally, Bayesian network inference revealed potential network mechanisms of EVL regulation.ConclusionsMonocyte differentiation-related prognostic signatures based on the Lasso + CoxBoost combination were able to accurately predict the prognostic status of patients with sepsis. In addition, low EVL expression was associated with poor prognosis in sepsis.

Single-cell Sequence Analysis Combined with Multiple Machine Learning to Identify Markers in Sepsis Patients: LILRA5.

Sepsis is a disease with a very high mortality rate, mainly involving an immune-dysregulated response due to bacterial infection. Most studies are currently limited to the whole blood transcriptome level; however, at the single cell level, there is still a great deal unknown about specific cell subsets and disease markers. We obtained 29 peripheral blood single-cell sequencing data, including 66,283 cells from 10 confirmed samples of sepsis infection and 19 healthy samples. Cells related to the sepsis phenotype were identified and characterized by the "scissor" method. The regulatory relationships of sepsis-related phenotype cells in the cellular communication network were clarified using the "cell chat" method. The least absolute shrinkage and selection operator (LASSO), support vector machine (SVM), and random forest (RF) were used to identify sepsis signature genes of diagnostic value. External validation was performed using multiple datasets from the GEO database (GSE28750, GSE185263, GSE57065) and 40 clinical samples. Bayesian algorithm was used to calculate the regulatory network of LILRA5 co-expressed genes. The stability of atenolol-targeting LILRA5 was determined by molecular docking techniques. Ultimately, action trajectory and survival analyses demonstrate the effectiveness of atenolol-targeted LILRA5 in treating patients with sepsis. We successfully identified 1215 healthy phenotypic cells and 462 sepsis phenotypic cells. We focused on 447 monocytes of the sepsis phenotype. Among the cellular communications, there were a large number of differences between these cells and other immune cells showing a significant inflammatory phenotype compared to the healthy phenotypic cells. Together, the three machine learning algorithms identified the LILRA5 marker gene in sepsis patients, and validation results from multiple external datasets as well as real-world clinical samples demonstrated the robust diagnostic performance of LILRA5. The AUC values of LILRA5 in the external datasets GSE28750, GSE185263, and GSE57065 could reach 0.875, 0.940, and 0.980, in that order. Bayesian networks identified a large number of unknown regulatory relationships for LILRA5 co-expression. Molecular docking results demonstrated the possibility of atenolol targeting LILRA5 for the treatment of sepsis. Behavioral trajectory analysis and survival analysis demonstrate that atenolol has a desirable therapeutic effect. LILRA5 is a marker gene in sepsis patients, and atenolol can stably target LILRA5.

A lightweight D2D authentication protocol for relay coverage scenario in 5G mobile network

5G cellular network is becoming a preferred communication network for deployment of the Internet of Things (IoT), due to its high speed, better connectivity, increased bandwidth, lower latency and flexibility. As a result, the traffic load of 5G cellular network is bound to increase manifold in the near future. Therefore, there is a requirement to explore the use of technologies that have the potential to ease the traffic load off the core network. Device-to-Device (D2D) communication in 5G cellular network for relay coverage is one such technology that enables two devices to communicate directly with each other through a local communication medium, even if one of them is not in the coverage area of the 5G cellular network. This technology has the potential to provide wider coverage, reduce the traffic load of the core network and minimize communication latency. For smooth D2D communication, the devices need to be authenticated. In technical specification-23.303 that is standardized by 3rd generation partnership project, basic guidelines for D2D communication are provided. However, in this specification no mechanism for authentication of the devices is proposed. It is also observed that in recent times a few authentication protocols have been proposed for D2D communication, but none of them are designed for relay coverage scenario in 5G cellular network. Hence, to fill the above discussed gap, in this paper we propose a lightweight D2D protocol for relay coverage scenario in 5G Mobile Network, that uses elliptic curve cryptography and symmetric key cryptography. To check the robustness of the proposed protocol, security analysis using formal verification techniques/tools like BAN logic, Scyther and AVISPA is done. The results of the analysis show the protocol to be safe against various attacks. Moreover, performance analysis of the protocol shows it to be lightweight and efficient, in terms of computation and communication cost, compared to other related works in the literature.

Video Stream Recognition Using Bitstream Shape for Mobile Network QoE.

Video streaming service delivery is a challenging task for mobile network operators. Knowing which services clients are using could help ensure a specific quality of service and manage the users' experience. Additionally, mobile network operators could apply throttle, traffic prioritization, or differentiated pricing. However, due to the growth of encrypted Internet traffic, it has become difficult for network operators to recognize the type of service used by their clients. In this article, we propose and evaluate a method for recognizing video streams solely based on the shape of the bitstream on a cellular network communication channel. To classify bitstreams, we used a convolutional neural network that was trained on a dataset of download and upload bitstreams collected by the authors. We demonstrate that our proposed method achieves an accuracy of over 90% in recognizing video streams from real-world mobile network traffic data.

Healthy and Osteoarthritis-Affected Joints Facing the Cellular Crosstalk.

Osteoarthritis (OA) is a chronic, progressive, severely debilitating, and multifactorial joint disease that is recognized as the most common type of arthritis. During the last decade, it shows an incremental global rise in prevalence and incidence. The interaction between etiologic factors that mediate joint degradation has been explored in numerous studies. However, the underlying processes that induce OA remain obscure, largely due to the variety and complexity of these mechanisms. During synovial joint dysfunction, the osteochondral unit undergoes cellular phenotypic and functional alterations. At the cellular level, the synovial membrane is influenced by cartilage and subchondral bone cleavage fragments and extracellular matrix (ECM) degradation products from apoptotic and necrotic cells. These "foreign bodies" serve as danger-associated molecular patterns (DAMPs) that trigger innate immunity, eliciting and sustaining low-grade inflammation in the synovium. In this review, we explore the cellular and molecular communication networks established between the major joint compartments-the synovial membrane, cartilage, and subchondral bone of normal and OA-affected joints.

CCN6 influences transcription and controls mitochondrial mass and muscle organization.

Mutations in Cellular Communication Network Factor 6 (CCN6) are linked to the debilitating musculoskeletal disease Progressive Pseudo Rheumatoid Dysplasia (PPRD), which disrupts mobility. Yet, much remains unknown about CCN6 function at the molecular level. In this study, we revealed a new function of CCN6 in transcriptional regulation. We demonstrated that CCN6 localizes to chromatin and associates with RNA Polymerase II in human chondrocyte lines. Using zebrafish as a model organism we validated the nuclear presence of CCN6 and its association with RNA Polymerase II in different developmental stages from 10 hpf embryo to adult fish muscle. In concurrence with these findings, we confirmed the requirement of CCN6 in the transcription of several genes encoding mitochondrial electron transport complex proteins in the zebrafish, both in the embryonic stages and in the adult muscle. Reduction in the expression of these genes upon morpholino-mediated knockdown of CCN6 protein expression led to reduced mitochondrial mass, which correlated with defective myotome organization during zebrafish muscle development. Overall, this study suggests that the developmental musculoskeletal abnormalities linked with PPRD could be contributed at least partly by impaired expression of genes encoding mitochondrial electron transport complexes due to defects in CCN6 associated transcriptional regulation.

Accurate Base Station Placement in 4G LTE Networks Using Multiobjective Genetic Algorithm Optimization

Cellular mobile communication network planning and optimization involve a complex engineering process that deals with network fundamentals, radio resource elements, and critical decision variables. The continuous evolution of radio access technologies provides new challenges that necessitate efficient radio planning and optimization. Therefore, the planning and optimization algorithms should be highly efficient, advanced, and robust. An important component of 4G LTE network planning is the proper placement of evolved node base stations (eNodeBs) and the configuration of their antenna elements. This contribution proposes a multiobjective genetic algorithm that integrates network coverage, capacity, and power consumption for optimal eNodeB placement in an operational 4G LTE network. The multi-objective-based genetic algorithm optimization has been achieved using the optimization toolbox in MATLAB. By leveraging the proposed method, the effect of different population sizes on the cost of placing the eNodeBs and the percentage coverage of the eNodeBs in a given cell is determined. As a result, the optimal selection technique that minimizes the total network cost without compromising the desired coverage and capacity benchmarks is achieved. The proposed automatic eNodeB antenna placement method can be explored to optimize 4G LTE cellular network planning in related wireless propagation environments.

Do not overwork: cellular communication network factor 3 for life in cartilage.

Cellular communication network factor (CCN) 3, which is one of the founding members of the CCN family, displays diverse functions. However, this protein generally represses the proliferation of a variety of cells. Along with skeletal development, CCN3 is produced in cartilaginous anlagen, growth plate cartilage and epiphysial cartilage. Interestingly, CCN3 is drastically induced in the growth plates of mice lacking CCN2, which promotes endochondral ossification. Notably, chondrocytes in these mutant mice with elevated CCN3 production also suffer from impaired glycolysis and energy metabolism, suggesting a critical role of CCN3 in cartilage metabolism. Recently, CCN3 was found to be strongly induced by impaired glycolysis, and in our study, we located an enhancer that mediated CCN3 regulation via starvation. Subsequent investigations specified regulatory factor binding to the X-box 1 (RFX1) as a transcription factor mediating this CCN3 regulation. Impaired glycolysis is a serious problem, resulting in an energy shortage in cartilage without vasculature. CCN3 produced under such starved conditions restricts energy consumption by repressing cell proliferation, leading chondrocytes to quiescence and survival. This CCN3 regulatory system is indicated to play an important role in articular cartilage maintenance, as well as in skeletal development. Furthermore, CCN3 continues to regulate cartilage metabolism even during the aging process, probably utilizing this regulatory system. Altogether, CCN3 seems to prevent "overwork" by chondrocytes to ensure their sustainable life in cartilage by sensing energy metabolism. Similar roles are suspected to exist in relation to systemic metabolism, since CCN3 is found in the bloodstream.