Perlecan/heparan sulfate proteoglycan 2 (HSPG2), a large HSPG, is indispensable for the development of musculoskeletal tissues, where it is deposited within the pericellular matrix (PCM) surrounding chondrocytes and disappears nearly completely at the chondro-osseous junction (COJ) of developing long bones. Destruction of perlecan at the COJ converts an avascular cartilage compartment into one that permits blood vessel infiltration and osteogenesis. Mutations in perlecan are associated with chondrodysplasia with widespread musculoskeletal and joint defects. This study elucidated novel signaling roles of perlecan core protein in endochondral bone formation and chondrocyte behavior. Perlecan subdomains were tested for chondrogenic properties in ATDC5 cells, a model for early chondrogenesis. A region within domain IV of perlecan (HSPG2 IV-3) was found to promote rapid prechondrocyte clustering. Introduction of the mutation (R3452Q) associated with the human skeletal disorder Schwartz-Jampel syndrome limited HSPG2 IV-3-induced clustering. HSPG2 IV-3 activity was enhanced when thermally unfolded, likely because of increased exposure of the active motif(s). HSPG2 IV-3-induced clustering was accompanied by the deactivation of key components of the focal adhesion complex, FAK and Src, with increased messenger RNA (mRNA) levels of precartilage condensation markers Sox9 and N-cadherin ( Cdh2), and cartilage PCM components collagen II ( Col2a1) and aggrecan ( Acan). HSPG2 IV-3 reduced signaling through the ERK pathway, where loss of ERK1/2 phosphorylation coincided with reduced FoxM1 protein levels and increased mRNA levels cyclin-dependent kinase inhibitor 1C (Cdkn1c) and activating transcription factor 3 ( Atf3), reducing cell proliferation. These findings point to a critical role for perlecan domain IV in cartilage development through triggering chondrocyte condensation.

The mechanistic aspects of a novel methodology for the electrochemical hydride generation (EcHG) of cadmium, entitled as the catholyte variation, have been studied. The hydrogen overvoltage of different cathode materials was determined in dilute electrolyte. The lead-tin alloy used as the cathode material for the EcHG of Cd had the highest hydrogen overpotential. In this technique, both electrolyte medium and cathode surface would change during electrolysis. The catholyte variation EcHG coupled with a flame heated quartz tube atomizer (QTA) atomic absorption spectrometer (AAS) produced peak-shaped transient signal of Cd. The analytical signal was significantly enhanced at neutralization time. The simultaneous production of lead and tin hydrides as well as the hydrogen at that time may speed up the desorption of cadmium species from the surface of the cathode. Also the high hydrogen overvoltage and the neutral medium aided the formation and separation of cadmium hydride molecules. The generated CdH2 vapour was kinetically stable.

Prior studies demonstrate that astrotactin (ASTN1) provides a neuronal receptor for glial-guided CNS migration. Here we report that ASTN1 binds N-cadherin (CDH2) and that the ASTN1:CDH2 interaction supports cell-cell adhesion. To test the function of ASTN1:CDH2 binding in glial-guided neuronal migration, we generated a conditional loss of Cdh2 in cerebellar granule cells and in glia. Granule cell migration was slowed in cerebellar slice cultures after a conditional loss of neuronal Cdh2, and more severe migration defects occurred after a conditional loss of glial Cdh2 Expression in granule cells of a mutant form of ASTN1 that does not bind CDH2 also slowed migration. Moreover, in vitro chimeras of granule cells and glia showed impaired neuron-glia attachment in the absence of glial, but not neuronal, Cdh2 Thus, cis and trans bindings of ASTN1 to neuronal and glial CDH2 form an asymmetric neuron-glial bridge complex that promotes glial-guided neuronal migration.

Hippocampal formation is important in spatial learning and memory. Members of the cadherin superfamily are observed in the neural system with diverse spatial and temporal expression patterns and are involved in many biological processes. To date, the avian hippocampal formation is not well understood. In this study, we examined the expression of cadherin mRNA in chicken and mouse brains to investigate the morphological and cytoarchitectural bases of hippocampal formation. Profiles of the spatiotemporal expression of cadherin mRNAs in the developing chicken embryonic parahippocampal area (APH) are provided, and layer-specific expression and spatiotemporal expression were observed in different subdivisions of the APH. That fact that some cadherins (Cdh2, Cdh8, Pcdh8 and Pcdh10) showed conserved regional expression both in the hippocampus and entorhinal cortex of mice and the hippocampal formation of chickens partially confirmed the structural homology proposed by previous scientists. This study indicates that some cadherins can be used as special markers of the avian hippocampal formation.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Normal gonad development assures the fertility of the individual. The properly functioning gonads must contain a sufficient number of the viable germ cells, possess a correct architecture and tissue structure, and assure the proper hormonal regulation. This is achieved by the interplay between the germ cells and different types of somatic cells. N-cadherin coded by the Cdh2 gene plays a critical role in this interplay. To gain an insight into the role of N-cadherin in the development of mouse gonads, we used the Cre-loxP system to knock out N-cadherin separately in two cell lines: the SF1⁺ somatic cells and the OCT4⁺ germ cells. We observed that N-cadherin plays a key role in the survival of both female and male germ cells. However, the N-cadherin is not necessary for the differentiation of the Sertoli cells or the initiation of the formation of testis cords or ovigerous cords. In the later stages of gonad development, N-cadherin is important for the maintenance of testis cord structure and is required for the formation of steroidogenic cells. In the ovaries, N-cadherin is necessary for the formation of the ovarian follicles. These results indicate that N-cadherin plays a major role in gonad differentiation, structuralization, and function.

The multiplex role of cadherin-based adhesion complexes during development of pallial excitatory neurons has been thoroughly characterized. In contrast, much less is known about their function during interneuron development. Here, we report that conditional removal of N-cadherin (Cdh2) from postmitotic neuroblasts of the subpallium results in a decreased number of Gad65-GFP-positive interneurons in the adult cortex. We also found that interneuron precursor migration into the pallium was already delayed at E14. Using immunohistochemistry and TUNEL assay in the embryonic subpallium, we excluded decreased mitosis and elevated cell death as possible sources of this defect. Moreover, by analyzing the interneuron composition of the adult somatosensory cortex, we uncovered an unexpected interneuron-type-specific defect caused by Cdh2-loss. This was not due to a fate-switch between interneuron populations or altered target selection during migration. Instead, potentially due to the migration delay, part of the precursors failed to enter the cortical plate and consequently got eliminated at early postnatal stages. In summary, our results indicate that Cdh2-mediated interactions are necessary for migration and survival during the postmitotic phase of interneuron development. Furthermore, we also propose that unlike in pallial glutamatergic cells, Cdh2 is not universal, rather a cell type-specific factor during this process.

Bovine sequence tagged sites (STSs) were developed for seven genes and used for synteny mapping with a hybrid bovine x rodent cell line panel. The genes were thymidylate synthase (TYMS), pituitary adenylate cyclase activating peptide (ADCYAP1), and melanocortin-2 receptor (MC2R) from the short arm of human chromosome (HSA) 18 and N-cadherin (CDH2), transthyretin (TTR), gastrin-releasing peptide (GRP), and plasminogen activator inhibitor 2 (PAI2) from the long arm of HSA 18. Primers for these genes were designed with human, ovine, or bovine sequences aligned with a sequence from a second species. The bovine PCR product was cloned, and the fragment was sequenced to verify that the homologous gene was indeed amplified. A second set of bovine-specific PCR primers were developed for each gene from these sequences. These STSs were used for synteny mapping, and all seven genes were syntenic with markers of bovine chromosome (BTA) 24. The concordance with BTA 24 was at least 96.5% for all genes.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a well-known immunotoxic environmental pollutant. However, most immunotoxicology studies of TCDD were based on the animal models and the inner mechanisms have just focused on a few genes/proteins. In this study, the immune functions of THP-1-derived macrophages was measured with in-vitro bioassays after 24-h exposure of TCDD including environmentally relevant concentrations. RNA-seq and Weighted Gene Co-expression Network Analysis were used to characterize the immunotoxicity molecular mechanisms. Our study is the first report on the TCDD-induced effects of cell adhesion, morphology, and multiple cytokines/chemokines production on THP-1 macrophages. After TCDD treatment, we observed an inhibited cell adherence, probably attributed to the suppressed mRNA levels of adhesion molecules ICAM-1, VCAM-1 and CD11b, and a decrease in cell pseudopodia and expression of F-actin. The inflammatory cytokines TNF-α, IL-10 and other 8 cytokines/chemokines regulating granulocytes/T cells and angiogenesis were disrupted by TCDD. Alternative splicing event was found to be a sensitive target for TCDD. Using WGCNA, we identified 10 hub genes (TNF, SRC, FGF2, PTGS2, CDH2, GNG11, BDNF, WNT5A, CXCR5 and RUNX2) highly relevant to these observed phenotypes, suggesting AhR less important in the effects TCDD have on THP-1 macrophages than in other cells. Our findings broaden the understanding of TCDD immunotoxicity on macrophages and provide new potential targets for clarifying the molecular mechanisms.

Keywords: Immunotoxicity; Macrophage; TCDD; Transcriptome; WGCNA.

Urokinase-type plasminogen activator (uPA; encoded by Plau) is a serine proteinase that, in the central nervous system, induces astrocytic activation. β-Catenin is a protein that links the cytoplasmic tail of cadherins to the actin cytoskeleton, thus securing the formation of cadherin-mediated cell adhesion complexes. Disruption of cell-cell contacts leads to the detachment of β-catenin from cadherins, and β-catenin is then degraded by the proteasome following its phosphorylation by GSK3β. Here, we show that astrocytes release uPA following a scratch injury, and that this uPA promotes wound healing via a plasminogen-independent mechanism. We found that uPA induces the detachment of β-catenin from the cytoplasmic tail of N-cadherin (NCAD; also known as CDH2) by triggering its phosphorylation at Tyr654. Surprisingly, this is not followed by degradation of β-catenin because uPA also induces the phosphorylation of the low density lipoprotein receptor-related protein 6 (LRP6) at Ser1490, which then blocks the kinase activity of GSK3β. Our work indicates that the ensuing cytoplasmic accumulation of β-catenin is followed by its nuclear translocation and β-catenin-triggered transcription of the receptor for uPA (Plaur), which in turn is required for uPA to induce astrocytic wound healing.

Keywords: Low density lipoprotein receptor-related protein 6; Plasmin; Urokinase-type plasminogen activator; Wnt-β-catenin pathway; uPA; β-catenin.

Objective: The aim of this study is the identification of hub genes associated with idiopathic pulmonary arterial hypertension (IPAH).

Materials and methods: GSE15197 gene expression data was downloaded from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) were identified by screening IPAH patients and controls. The 5,000 genes with the greatest variances were analyzed using a weighted gene co-expression network analysis (WGCNA). Modules with the strongest correlation with IPAH were chosen, followed by a functional enrichment analysis. Protein-protein interaction (PPI) networks were constructed to identify hub gene candidates using calculated degrees. Real hub genes were found from the overlap of DEGs and candidate hub genes. microRNAs (miRNAs) targeting real hub genes were found by screening miRNet 2.0. The most important IPAH miRNAs were identified.

Results: There were 4,395 DEGs identified. WGCNA indicated that green and brown modules associated most strongly with IPAH. Functional enrichment analysis showed that green and brown module genes were mainly involved in protein digestion and absorption and proteoglycans in cancer, respectively. The top ten candidate hub genes in green and brown modules were identified, respectively. After overlapping with DEGs, 11 real hub genes were identified: EP300, MMP2, CDH2, CDK2, GNG10, ALB, SMC2, DHX15, CUL3, BTBD1, and LTN1. These genes were expressed with significant differences in IPAH versus controls, indicating a high diagnostic ability. The miRNA-gene network showed that hsa-mir-1-3p could associate with IPAH.

Conclusion: EP300, MMP2, CDH2, CDK2, GNG10, ALB, SMC2, DHX15, CUL3, BTBD1, and LTN1 may play essential roles in IPAH. Predicted miRNA hsa-mir-1-3p could regulate gene expression in IPAH. Such hub genes may contribute to the pathology and progression in IPAH, providing potential diagnostic and therapeutic opportunities for IPAH patients.

Keywords: differentially expressed genes; functional enrichment analysis; hub genes; idiopathic pulmonary arterial hypertension; weighted gene co-expression network analysis.

Inhibitors of DNA binding and cell differentiation (ID) proteins regulate cellular differentiation and tumor progression. Whether ID family proteins serve as a linkage between pathological differentiation and cancer stemness in colorectal cancer is largely unknown. Here, the expression of ID4, but not other ID family proteins, was enriched in LGR5-high colon cancer stem cells. Its high expression was associated with poor pathological differentiation of colorectal tumors and shorter survival in patients. Knockdown of ID4 inhibited the growth and dissemination of colon cancer cells, while enhancing chemosensitivity. Through gene expression profiling analysis, brain-derived neurotrophic factor (BDNF) was identified as a downstream target of ID4 expression in colorectal cancer. BDNF knockdown decreased the growth and migration of colon cancer cells, and its expression enhanced dissemination, anoikis resistance, and chemoresistance. ID4 silencing attenuated the epithelial-to-mesenchymal transition (EMT) pattern in colon cancer cells. Gene cluster analysis revealed that ID4 and BDNF expression was clustered with mesenchymal markers and distant from epithelial genes. BDNF silencing decreased the expression of mesenchymal markers Vimentin, CDH2 and SNAI1. These findings demonstrated that ID4-BDNF signaling regulates colorectal cancer survival, with the potential to serve as a prognostic marker in colorectal cancer.

Endothelial cells sense and respond to shear stress. Different in vitro model systems have been used to study the cellular responses to shear stress, but these platforms do not allow studies on high numbers of cells under uniform and controllable shear stress. The annular dish, or dish-in-a-dish (DiaD), on the orbital shaker has been proposed as an accessible system to overcome these challenges. However, the influence of the DiaD design and the experimental parameters on the shear stress patterns is not known. In this study, we characterize different designs and experimental parameters (orbit size, speed and fluid height) using computational fluid dynamics. We optimize the DiaD for an atheroprotective flow, combining high shear stress levels with a low oscillatory shear index (OSI). We find that orbit size determines the DiaD design and parameters. The shear stress levels increase with increasing rotational speed and fluid height. Based on our optimization, we experimentally compare the 134/56 DiaD with regular dishes for cellular alignment and KLF2, eNOS, CDH2 and MCP1 expression. The calculated OSI has a strong impact on alignment and gene expression, emphasizing the importance of characterizing shear profiles in orbital setups.

Keywords: computational fluid dynamics; endothelial cells; flow; orbital shaker; shear stress.

In this article, we propose a systemic approach to investigate the impact of electrohydrodynamic jetting (EHDJ) encapsulation on viability, proliferation, and functionality of the encapsulated cells. EHDJ consists in applying a high-voltage electrical field between a target substrate and a jetting needle, which is fed with a suspension of cells in a polymeric solution undergoing a sol-gel transition upon contact with the target. The viability, proliferation, and self-assembling ability of SHSY5Y human neuroblastoma cell line encapsulated in 2% alginate microbeads were analyzed by confocal microscopy and DNA quantification assays. In addition, the expression of stress (HSP70B'), apoptotic (CASP3), necrotic (HMGB1), hypoxic (HYOU1, GAPDH), and adhesion (CDH2) markers was measured with reverse transcription quantitative polymerase chain reaction (qPCR). After an initial upregulation of the HSP70B' expression within 24 h, its expression decreased to the negative control level together with a decrease in the expression of CASP3. Any increase in necrotic or hypoxic marker expression was not detected, while a slight upregulation of CDH2 was observed in the first days after encapsulation, followed by its downregulation and stabilization to the control level. Furthermore, cell-laden beads started to self-assemble in three-dimensional (3D) constructs from the 3rd week after encapsulation. The results indicated that the EHDJ encapsulation method had a mild effect on cells, which after a week, fully recovered their proliferation rate and ability to self-assemble into 3D constructs.

The mechanism underlying increased concentrations of cancer stem cell (CSC)-associated factors in non-small cell lung cancer (NSCLC) cells treated with transforming growth factor β1 (TGFβ1) and tumor necrosis factor α (TNFα), is still not clear. The purpose of this study was to investigate the possible role of CD44 in the regulation of CSC-associated genes, by analyzing the effect of CD44 knockdown on their expression. A549, a NSCLC cell line that expresses CD44 antigen, was treated with TGFβ1 and TNFα. Small-interfering ribonucleic acid (siRNA) that specifically targets the CD44 gene was used to knockdown the expression of CD44 in A549. The gene expressions of CD44, CXCR4, POU5F1 (octamer-binding transcription factor 4 [Oct4]), PROM1, NANOG, c-Myc, KLF4, and SOX2, as well as of CDH1 (E-cadherin), CDH2 (N-cadherin), VIM (vimentin), and FN1 (fibronectin) were analyzed in A549 cells by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Cell morphology was observed using light microscopy. After TGFβ1/TNFα treatment, increased expressions of CXCR4 and POU5F1 were detected. Silencing of CD44 gene expression was confirmed by RT-qPCR. The knockdown of CD44 decreased the CXCR4 and POU5F1 gene expressions in TGFβ1/TNFα-treated A549 cells. However, the silencing of CD44 did not affect the morphology of TGFβ1/TNFα-treated A549 cells nor it reversed epithelial-mesenchymal transition (EMT) gene signature induced by TGFβ1/TNFα in A549 cells. Our preliminary findings suggest that the CD44 gene may have a role in regulating CXCR4 and POU5F1 gene expressions, independently of the EMT signaling pathway.

Cadherins form connection between cells, facilitate communication, and serve as essential agents in the progression of multiple cancers. Over 100 cadherins have been identified and they are mainly divided into four groups: classical cadherins (CDHs), protocadherins (PCDHs), desmosomal (DSC), and cadherin-related proteins. Accumulating evidence has indicated that several members of the cadherins are involved in breast cancer development. Nevertheless, the expression profiles and corresponding prognostic outcomes of these breast cancer-related cadherins are yet to be analyzed. Here, we examined the expression levels and prognostic potential of these breast cancer-related cadherins from the specific databases viz. oncomine, gene expression profiling interactive analysis, human protein atlas, UALCAN, Kaplan-Meier Plotter, and cBioPortal. We found that the <em>CDH2</em>/11 levels were higher in breast cancer tissues, compared to healthy breast tissues, whereas with CDH3-5, PCDH8/10, and DSC3, the levels were lower in the former than in the latter. Additionally, for CDH1/6/13/17/23, PCDH7, and FAT4, trancript level alterations between breast cancer and healthy tissues varied across different databases. The CDH1 protein levels were elevated in breast cancer tissues versus healthy breast tissues, whereas the protein levels of CDH3/11 and PCDH8/10 were reduced in breast cancer, compared to healthy breast tissues. For CDH15 and <em>CDH2</em>3, the expression levels paralleled tumor stage. Survival analysis, using the Kaplan-Meier Plotter database, demonstrated that elevated CDH1-3 levels correlated with diminished relapse-free survival in breast cancer patients. Alternately, enhanced CDH4-6/15/17/23, PCDH10, DSC3, and FAT4 levels estimated a rise in relapse-free survival of breast cancer patients. These data suggest CDH1-3 to be a promising target for breast cancer precision therapy and CDH4-6/15/17/23, PCDH10, DSC3, and FAT4 to be novel biomarkers for breast cancer prognosis.

Keywords: Breast cancer; bioinformatics; biomarkers; cadherin; expression.

Tunneling chemical reactions between deuterated methyl radicals and the hydrogen molecule in a parahydrogen crystal have been studied by Fourier transform infrared spectroscopy. The tunneling rates of the reactions R + H2 ->> RH + H (R = CD3,CD2H,CDH2) in the vibrational ground state were determined directly from the temporal change in the intensity of the rovibrational absorption bands of the reactants and products in each reaction in solid parahydrogen observed at 5 K. The tunneling rate of each reaction was found to differ definitely depending upon the degree of deuteration in the methyl radicals. The tunneling rates were determined to be 3.3 x 10(-6) s(-1), 2.0 x 10(-6) s(-1), and 1.0 x 10(-6) s(-1) for the systems of CD3, CD2H, and CDH2, respectively. Conversely, the tunneling reaction between a CH3 radical and the hydrogen molecule did not proceed within a week's time. The upper limit of the tunneling rate of the reaction of the CH3 radical was estimated to be 8 x 10(-8) s(-1).

Objectives: Oral squamous cell carcinoma (OSCC) is a common head and neck cancer with high incidence. Numerous reports have elucidated that long non-coding RNAs (lncRNAs) serve as crucial regulatory factors in various diseases including cancer. Nonetheless, the role of lncRNA JPX in OSCC is still not elaborated.

Subjects and methods: In our research, we detected the expression of lncRNA JPX through qRT-PCR. Colony formation, EdU staining, Transwell, TUNEL and caspase-3 activity assays were applied for estimating the function of lncRNA JPX in OSCC. RIP, pull down and luciferase reporter experiments were adopted to examine the interrelations between lncRNA JPX, miR-944 and CDH2.

Results: LncRNA JPX was discovered as highly expressed in OSCC cells. Silencing lncRNA JPX restrained OSCC cell proliferation, migration and invasion. Interestingly, lncRNA JPX bound with miR-944, and then augmented CDH2 via a competing endogenous RNA (ceRNA) mechanism. Importantly, overexpressed CDH2 recovered the suppression of silenced lncRNA JPX on the oncogenic behaviors of OSCC cells.

Conclusions: In short, lncRNA JPX contributes to OSCC cell proliferation, migration and invasion via miR-944/CDH2 axis, which offers a new direction for potential targeted treatment of OSCC.

Keywords: CDH2; lncRNA JPX; miR-944; oral squamous cell carcinoma.

N-cadherin is a homophilic cell adhesion molecule that stabilizes excitatory synapses, by connecting pre- and post-synaptic termini. Upon NMDA receptor (NMDAR) activation by glutamate, membrane-proximal domains of N-cadherin are cleaved serially by a-disintegrin-and-metalloprotease 10 (ADAM10) and then presenilin 1(PS1, catalytic subunit of the γ-secretase complex). To assess the physiological significance of the initial N-cadherin cleavage, we engineer the mouse genome to create a knock-in allele with tandem missense mutations in the mouse N-cadherin/Cadherin-2 gene (Cdh2 ^{R714G, I715D}, or GD) that confers resistance on proteolysis by ADAM10 (GD mice). GD mice showed a better performance in the radial maze test, with significantly less revisiting errors after intervals of 30 and 300 s than WT, and a tendency for enhanced freezing in fear conditioning. Interestingly, GD mice reveal higher complexity in the tufts of thorny excrescence in the CA3 region of the hippocampus. Fine morphometry with serial section transmission electron microscopy (ssTEM) and three-dimensional (3D) reconstruction reveals significantly higher synaptic density, significantly smaller PSD area, and normal dendritic spine volume in GD mice. This knock-in mouse has provided in vivo evidence that ADAM10-mediated cleavage is a critical step in N-cadherin shedding and degradation and involved in the structure and function of glutamatergic synapses, which affect the memory function.

Keywords: ADAM10; Hippocampus; N-cadherin; Synapse; Working memory.

Introduction: Heterogeneity of non-small cell lung carcinoma (NSCLC) among patients is currently not well studied. Pathologic markers and staging systems have not been a precise predictor of the prognosis of an individual patient. Hence, we hypothesize to develop a transcript-based signature to categorize stage IIIA-NSCLC in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC), plus identify markers that could indicate the prognosis of the disease.

Methods: Human Transcriptome Array 2.0 (HTA) and NanoString nCounter® platform were used for high-throughput gene-expression profiling. Initially, we profiled stage IIIA-NSCLC through HTA and validated through NanoString. Additionally, two metastatic markers SPP1 and CDH2 were validated in 47 NSCLC stage IIIA samples through real-time PCR.

Results: We observed distinct gene clusters in LUAD and LUSC with down-regulation of six genes and up-regulation of 57 genes through HTA. Ninety-six transcripts were randomly selected after analyzing HTA data and validated on the NanoString platform. We found 40 differentially expressed transcripts that categorized NSCLC into LUAD and LUSC. SPP1 is significantly overexpressed (4.311±1.27 fold in LUAD and 13.41±3.82 fold in LUSC compared to control), and the CDH2 transcript was significantly overexpressed (11.53 ± 4.027-fold compared to control) only in LUSC.

Discussion: These markers enable us to categorize stage IIIA NSCLC into LUAD and LUSC plus these markers may be helpful to understand the pathophysiology of NSCLC. However, more data required to make these findings useful in general clinical practice.

Keywords: NanoString; human transcriptome array; lung adenocarcinoma; lung squamous cell carcinoma; non-small cell lung cancer.

Among the genetic factors playing a key role in the etiology of intellectual disabilities (IDs) and autism spectrum disorders (ASDs), several encode RNA-binding proteins (RBPs). In this study, we deciphered the molecular and cellular bases of ID-ASD in a patient followed from birth to the age of 21, in whom we identified a de novo CSDE1 (Cold Shock Domain-containing E1) nonsense variation. CSDE1 encodes an RBP that regulates multiple cellular pathways by monitoring the translation and abundance of target transcripts. Analyses performed on the patient's primary fibroblasts showed that the identified CSDE1 variation leads to haploinsufficiency. We identified through RNA-seq assays the Wnt/β-catenin signaling and cellular adhesion as two major deregulated pathways. These results were further confirmed by functional studies involving Wnt-specific luciferase and substrate adhesion assays. Additional data support a disease model involving APC Down-Regulated-1 (APCDD1) and cadherin-2 (CDH2), two components of the Wnt/β-catenin pathway, CDH2 being also pivotal for cellular adhesion. Our study, which relies on both the deep phenotyping and long-term follow-up of a patient with CSDE1 haploinsufficiency and on ex vivo studies, sheds new light on the CSDE1-dependent deregulated pathways in ID-ASD.

The expression of DNA repair (DDB1, ERCC4, ERCC5), leukocyte adhesion (VCAM1, ICAM1, SELE, SELP), endothelial mechanotransduction (KLF4), endothelial differentiation (PECAM1, CDH5, CD34, NOS3), endothelial-to-mesenchymal transition (SNAI1, SNAI2, TWIST1, GATA4, ZEB1, CDH2), scavenger receptors (LOX1, SCARF1, CD36, LDLR, VLDR), antioxidant system (PXDN, CAT, SOD1) and transcription factor (HEY2) genes in primary human coronary (HCAEC) and internal thoracic (HITAEC) arteries endothelial cells exposed to alkylating mutagen mitomycin C (MMC) was studied at two time points - after 6 h of incubation with MMC and after 6 h of the genotoxic load followed by 24 h of incubation in pure culture medium using the quantitative PCR. Immediately after MMC exposure, in the exposed HCAEC and HITAEC a decreased expression of almost all studied genes was noted excepted SNAI, which demonstrated a 4-told increase in its expression compared to the unexposed control. Elimination of MMC from the cultures, an increased expression of the VCAM1, ICAM1, SELE, SNAI2, KLF4 genes and a decreased the mRNA level of the PECAM1, CDH5, CD34, ZEB1, CAT, PXDN genes were observed in both cell lines. In addition, HITAEC cells were characterized by a decreased expression of the SOD1, SCARF1, CD36 genes and an increased expression of the SNAI1 and TWIST1 genes; in HCAEC, an increased mRNA level of the LDLR and VLDLR genes was noted. Thus, MMC-induced genotoxic stress is associated with the endothelial dysfunction.

Provedena otsenka ékspressii genov reparatsii DNK (DDB1, ERCC4, ERCC5), adgezii leĭkotsitov (VCAM1, ICAM1, SELE, SELP), éndotelial'noĭ mekhanotransduktsii (KLF4), éndotelial'noĭ differentsirovki (PECAM1, CDH5, CD34, NOS3), éndotelial'no-mezenkhimal'nogo perekhoda (SNAI1, SNAI2, TWIST1, GATA4, ZEB1, CDH2), skavendzher-retseptorov (LOX1, SCARF1, CD36, LDLR, VLDR), antioksidantnoĭ zashchity (PXDN, CAT, SOD1) i transkriptsionnogo faktora HEY2 v kul'turakh pervichnykh éndotelial'nykh kletok koronarnoĭ (HCAEC) i vnutrenneĭ grudnoĭ (HITAEC) arteriĭ cheloveka, éksponirovannykh alkiliruiushchim mutagenom mitomitsinom S (MMS). Uroven' ékspressii opredeliali s pomoshch'iu metoda kolichestvennoĭ PTsR v dvukh vremennykh tochkakh: cherez 6 ch kul'tivirovaniia kletok v prisutstvii MMS i cherez 6 ch ékspozitsii mutagenom s posleduiushchimi 24 ch kul'tivirovaniia v chistoĭ kul'tural'noĭ srede. Neposredstvenno posle vozdeĭstviia mutagena v éksponirovannykh kul'turakh HCAEC i HITAEC bylo otmecheno snizhenie ékspressii prakticheski vsekh izuchennykh genov, za iskliucheniem SNAI2 (ego ékspressiia byla povyshena v 4 raza) po sravneniiu s neéksponirovannym kontrolem. Posle éliminirovaniia iz kul'tur MMS v obeikh kletochnykh liniiakh bylo otmecheno uvelichenie ékspressii genov VCAM1, ICAM1, SELE, SNAI2, KLF4 i snizhenie ékspressii genov PECAM1, CDH5, CD34, ZEB1, CAT, PXDN. Krome togo, kletki HITAEC takzhe kharakterizovalis' snizheniem ékspressii genov SOD1, SCARF1, CD36 i povysheniem ékspressii genov SNAI1 i TWIST1; v kletkakh HCAEC bylo otmecheno uvelichenie ékspressii genov LDLR i VLDLR. Takim obrazom, genotoksicheskiĭ stress, vyzvannyĭ mutagenom alkiliruiushchego mekhanizma deĭstviia, assotsiirovan s razvitiem éndotelial'noĭ disfunktsii, chto proiavliaetsia v izmenenii profilia gennoĭ ékspressii éndotelial'nykh kletok, kul'tiviruemykh v usloviiakh genotoksicheskoĭ nagruzki.

Keywords: atherogenesis; endothelial dysfunction; gene expression; mutagenesis.

The challenge for treating breast cancer (BC) is partly due to long-term dormancy driven by cancer stem cells (CSCs) capable of evading immune response and resist chemotherapy. BC cells show preference for the BM, resulting in poor prognosis. CSCs use connexin 43 (Cx43) to form gap junctional intercellular communication with BM niche cells, fibroblasts, and mesenchymal stem cells (MSCs). However, Cx43 is an unlikely target to reverse BC dormancy because of its role as a hematopoietic regulator. We found N-cadherin (CDH2) and its associated pathways as potential drug targets. CDH2, highly expressed in CSCs, interacts intracellularly with Cx43, colocalizes with Cx43 in BC cells within BM biopsies of patients, and is required for Cx43-mediated gap junctional intercellular communication with BM niche cells. Notably, CDH2 and anti-apoptotic pathways maintained BC dormancy. We thereby propose these pathways as potential pharmacological targets to prevent dormancy and chemosensitize resistant CSCs.

Pesticides despite being agents that protect the plants and humans from noxious pests, are infamous for their potential to cause detrimental health issues in nontargeted species. In order to ascertain the latter, a set of experiments were conducted by exposing early chick embryos to a widely used combination insecticide (Ci, 50% chlorpyrifos and 5% cypermethrin). The results revealed a myriad of congenital defects pertaining to craniofacial development such as anophthalmia, microphthalmia, exencephaly as well as deformed beak and cranial structures. These teratological manifestations could be attributed to the Ci induced alteration in the titre of major regulators of neurulation and ossification. Therefore, the mRNA and/or the protein level expression pattern of genes which are reported to be involved in the craniofacial development were studied at selected time points of embryonic development. The analysis of the result showed that there have been significant alternations in the expression patterns of the signalling molecules such as SHH, WNTs, CDH1, CDH2, L1CAM, PAX6, HOX, PCNA, GLI3, BMP7, FGF8, GLIs, SOX9, RUNX2, DLX5, COL10A1, CASPASE3 etc. on embryonic days 2, 4 and/or 10. Concurrently, on day 10, whole-mount skeletal staining and biochemical estimation of hydroxyproline were carried out in the cranial tissues of the embryos. The overall result of the current study indicates that exposure to Ci during early development impede the crucial regulatory signals that orchestrate the morphogenesis of cranial neural crest cells thereby hindering the normal progression of neural tube and endochondral ossification which collectively lead to craniofacial dysmorphism in domestic chicks.