bluuebunny/crossref_metadata_2025_split huggingface dataset:
Novel Role for Claudin-11 in the Regulation of Osteoblasts via Modulation of ADAM10-Mediated Notch Signaling
Authors: Lindsey Richard C , Xing Weirong , Pourteymoor Sheila , Godwin Catrina , Gow Alexander , Mohan Subburaman 
Published: May 2019

ABSTRACT
The claudin (Cldn) family comprises 27 members of 20 to 34 kDa transmembrane tight junction proteins. In addition to Cldns’ established canonical role as barriers controlling paracellular flow of molecules, a distinct noncanonical role for them as mediators of cell signaling is now emerging. In our studies evaluating Cldn family expression levels during osteoblast differentiation, Cldn-11 showed the largest increase (60-fold). Immunohistochemistry studies revealed high Cldn-11 expression in trabecular (Tb) bone lining cells. Micro-CT analysis of femurs and vertebrae of Cldn-11 knock-out (KO) mice at 12 weeks of age exhibited a 40% (p &lt; 0.01) reduction in Tb bone volume adjusted for tissue volume compared with control mice, a change caused by significant reductions in Tb number and thickness and increase in Tb separation. Histomorphometry and serum biomarker studies revealed that reduced bone formation, not increased resorption, is the cause for reduced Tb bone volume in the Cldn-11 KO mice. Cldn-11 KO osteoblasts expressed reduced ALP and BSP, whereas Cldn-11 overexpression in MC3T3-E1 cells increased expression of ALP and BSP. Mechanistically, Cldn-11 interacted with tetraspanin (Tspan)3 in osteoblasts, and Tspan3 knockdown reduced osteoblast differentiation. Because members of the Tspan family regulate cell functions via Notch signaling, we evaluated whether Cldn-11/Tspan3 regulates Notch signaling in osteoblasts. Accordingly, Notch targets Hey1 and Hey2 were significantly upregulated in Cldn-11 overexpressing cultures but downregulated in both Cldn-11 KO and Tspan3 knockdown osteoblasts. Because ADAM10 has been shown to interact with Tspan family members to regulate Notch signaling, we evaluated whether Cldn-11 regulates ADAM10 expression. Cldn-11 overexpressing cells express more mature ADAM10, and an ADAM10 inhibitor blocked the Cldn-11 effect on osteoblast differentiation. Based on these data, we propose Cldn-11 as a novel component of an osteoblast cell surface protein complex, comprising Tspan3 and ADAM10, which regulates Notch signaling and cell differentiation. © 2019 American Society for Bone and Mineral Research.
Knockdown of lncRNA‐UCA1 inhibits cell viability and migration of human glioma cells by miR‐193a‐mediated downregulation of CDK6
Authors: Xin Haibin , Liu Nina , Xu Xiaosheng , Zhang Jinwu , Li Yu , Ma Yongchao , Li Guoqiang , Liang Junjun 
Published: May 2019

AbstractBackgroundLong noncoding RNA urothelial carcinoma‐associated 1 (lncRNA‐UCA1) is generally recognized as an oncogenic molecule in several human malignant tumors. However, the available evidence does not necessarily imply an unequivocal causal function of UCA1 in glioblastoma. The current study was aimed to probe the biological function of lncRNA‐UCA1 in human glioblastoma cell lines. Besides, we further investigated the potential mechanisms.MethodsCell viability, apoptosis, as well as migration and invasion were measured using a commercial cell counting kit‐8, flow cytometry, and 24‐Transwell assay, respectively. LncRNA‐UCA1, microRNA‐193a (miR‐193a), and CDK6 at messenger RNA levels were evaluated by quantitative real‐time polymerase chain reaction method. Protein level was examined by Western blot analysis. RNA immunoprecipitation was utilized to validate lncRNA‐UCA1 associated with miR‐193a. Luciferase activity assay was used to identify the miR‐193a‐targeted CDK6 3′‐untranslated region.ResultslncRNA‐UCA1 knockdown weakened cell viability, augmented apoptosis progression, as well as suppressed migration and invasion behaviors in glioblastoma cells, whereas lncRNA‐UCA1 silence exhibited the opposite functions. lncRNA‐UCA1 functioned as an endogenous sponge of miR‐193a. miR‐193a silence reversed the biological function of lncRNA‐UCA1 knockdown on U‐118 MG cells. miR‐193a negatively regulated the expression of CDK6, and it affected the U‐118 MG cells through regulating CDK6 expression. CDK6 overexpression abrogated the blockage of PI3K/AKT, mitogen‐activated protein kinase (MAPK), and Notch signaling pathways. Furthermore, lncRNA‐UCA1 and miR‐193a could affect these signaling cascades through regulating CDK6 expression. The regulatory mechanisms of lncRNA‐UCA1 were further consolidated in clinical specimens.ConclusionlncRNA‐UCA1 silence reduced cell viability, promoted apoptosis progression, while impeding the migration and invasion of glioblastoma cells by miR‐193a‐mediated silence of CDK6, with blockage of PI3K/AKT, MAPK, and Notch pathways.
microRNA‐96 promotes osteoblast differentiation and bone formation in ankylosing spondylitis mice through activating the Wnt signaling pathway by binding to SOST
Authors: Ma Shu , Wang Dan‐Dan , Ma Cheng‐Yuan , Zhang Yan‐Dong 
Published: May 2019

AbstractAnkylosing spondylitis (AS) refers to a type of arthritis manifested with chronic inflammation of spine joints. microRNAs (MiRNAs) have been identified as new therapeutic targets for inflammatory diseases. In this study, we evaluated the influence of microRNA‐96 (miR‐96) on osteoblast differentiation together with bone formation in a murine model of AS. The speculated relationship that miR‐96 could bind to sclerostin (SOST) was verified by dual luciferase reporter assay. After successful model establishment, the mice with AS and osteoblasts isolated from mice with AS were treated with mimics or inhibitors of miR‐96, or DKK‐1 (a Wnt signaling inhibitor). The effects of gain‐ or loss‐of‐function of miR‐96 on the inflammatory cytokine release (IL‐6, IL‐10, and TNF‐α), alkaline phosphatase (ALP) activity, calcium nodule formation, along with the viability of osteoblasts were determined. It was observed that miR‐96 might target and regulate SOST. Besides, miR‐96 was expressed at a high level in AS mice while SOST expressed at a low level. TOP/FOP‐Flash luciferase reporter assay confirmed that miR‐96 activated the Wnt signaling pathway. Moreover, AS mice overexpressing miR‐96 exhibited increased contents of IL‐6, IL‐10 and TNF‐α, ALP activity, calcium nodule numbers, and viability of osteoblasts. In contrast, inhibition of miR‐96 resulted in suppression of the osteoblast differentiation and bone formation. In conclusion, the study implicates that overexpressing miR‐96 could improve osteoblast differentiation and bone formation in AS mice via Wnt signaling pathway activation, highlighting a potential new target for AS treatment.
Low glucose availability alters the expression of genes involved in initial adhesion of human glioblastoma cancer cell line SF767
Authors: Aftab Saira , Shakoori Abdul Rauf 
Published: May 2019

AbstractStudying the metabolic pathways of cancer cells is considered as a key to control cancer malignancies and open windows for effective drug discovery against cancer. Of all the properties of a tumor, metastasis potential is a defining characteristic. Metastasis is controlled by a variety of factors that directly control the expression of cell adhesion proteins. In this study we have investigated the expression of cell to cell and cell to matrix adhesion protein genes during the initial phases of attachment of human glioblastoma cancer cell line SF767 (66Y old human female: UCSF Neurosurgery Tissue Bank) to the attachment surface under (Cell culture treated polystyrene plate bottom) glucose‐rich and glucose‐starved conditions. The aim was to imitate the natural microenvironment of glucose availability to cancer cells inside a tumor that triggers epithelial to mesenchymal transition (EMT). In this study, we have observed the gene expression of epithelial and mesenchymal isoforms of cadherin (E‐CAD and N‐CAD) and Ig like cell adhesion molecules (E‐CAM and N‐CAM) along with Integrin family subunits for the initial attachment of cancer cells. We observed that high glucose environments promoted cell survival and cell adhesion, whereas low glucose accelerated EMT by downregulating the expression level of integrin, E‐CAD, and N‐CAD, and upregulation of N‐CAM during early period of cell adhesion. Low glucose availability also downregulated variety of structural and regulatory genes, such as zinc finger E‐box binding home box 1A), cytokeratin, Snail, and β catenin, and upregulation of hypoxia‐inducible factor 1, matrix metalloprotease 13/Collagenase 3, vimentim, p120, and fructose 1,6 bisphosphatase. Glucose conditions are more efficient for cancer studies in this case glioblastoma cells.
Long noncoding RNA MALAT‐1 inhibits apoptosis and matrix metabolism disorder in interleukin‐1β‐induced inflammation in articular chondrocytes via the JNK signaling pathway
Authors: Gao Gui‐Cheng , Cheng Xi‐Gao , Wei Qiang‐Qiang , Chen Wei‐Cai , Huang Wen‐Zhou 
Published: May 2019

AbstractProinflammatory cytokine such as interleukin (IL)‐1β causes inflammation of articular cartilage. In this current study, we explored the chondroprotective effects of long noncoding RNA (lncRNA) MALAT‐1 on cell proliferation, apoptosis, and matrix metabolism in IL‐1β‐induced inflammation in articular chondrocytes. Articular chondrocytes from knee joints of normal rats were isolated and cultured, followed by identification through observation of toluidine blue and COL II immunocytochemical stainings. The proliferation of chondrocytes at passage 2 was detected by the 3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyl‐2H‐tetrazolium bromide (MTT) assay. The inflammatory chondrocytes induced by 10 ng/mL IL‐1β were observed and identified by toluidine blue and COL II immunocytochemical stainings. pcDNA 3.1 and pcDNA‐MALAT‐1 were transfected in the chondrocytes. Ultrastructure of chondrocytes was observed by using a transmission electron microscope. The MTT assay was carried out to evaluate chondrocyte viability. Hoechst 33258 staining and flow cytometry were adopted to assess chondrocyte apoptosis. The chondrocytes at passage 2 with the biological characteristics of chondrocytes were used for subsequent experiments. In IL‐1β‐treated chondrocytes, the growth rate of chondrocytes slowed down, the cells became narrow and long, the vacuoles were seen in the cells, and the morphology of the chondrocytes was irregular. The toluidine blue staining and the immunohistochemical staining of COL II became weaker. In response to IL‐1β induction, articular chondrocytes showed reduced MALAT‐1 expression; moreover, obvious cartilage injury was observed with decreased chondrocyte viability and Col II expression and elevated chondrocyte apoptosis, MMP‐13 expression, and p‐JNK expression. With the treatment of pcDNA‐MALAT‐1, the cartilage injury was alleviated with increased chondrocyte viability and type II collagen (Col II) expression and reduced chondrocyte apoptosis, MMP‐13 expression and p‐JNK expression. Taken together these results, lncRNA MALAT‐1 blocked the activation of the JNK signaling pathway; thereby, IL‐1β‐induced inflammation in articular chondrocytes was reduced with enhanced chondrocyte proliferation and suppressed chondrocyte apoptosis and extracellular matrix degradation.
Novel c.G630A TCIRG1 mutation causes aberrant splicing resulting in an unusually mild form of autosomal recessive osteopetrosis
Authors: Zirngibl Ralph A. , Wang Andrew , Yao Yeqi , Manolson Morris F. , Krueger Joerg , Dupuis Lucie , Mendoza‐Londono Roberto , Voronov Irina 
Published: May 2019

AbstractAutosomal recessive osteopetrosis (ARO) is a severe genetic bone disease characterized by high bone density due to mutations that affect formation or function of osteoclasts. Mutations in the a3 subunit of the vacuolar‐type H+‐ATPase (encoded by T‐cell immune regulator 1 [TCIRG1]) are responsible for ~50% of all ARO cases. We identified a novel TCIRG1 (c.G630A) mutation responsible for an unusually mild form of the disease. To characterize this mutation, osteoclasts were differentiated using peripheral blood monocytes from the patient (c.G630A/c.G630A), male sibling (+/+), unaffected female sibling (+/c.G630A), and unaffected parent (+/c.G630A). Osteoclast formation, bone‐resorbing function, TCIRG1 protein, and mRNA expression levels were assessed. The c.G630A mutation did not affect osteoclast differentiation; however, bone‐resorbing function was decreased. Both TCIRG1 protein and full‐length TCIRG1 mRNA expression levels were also diminished in the affected patient's sample. The c.G630A mutation replaces the last nucleotide of exon 6 and may cause splicing defects. We analyzed the TCIRG1 splicing pattern between exons 4 to 8 and detected deletions of exons 5, 6, 7, and 5‐6 (ΔE56). These deletions were only observed in c.G630A/c.G630A and +/c.G630A samples, but not in +/+ controls. Among these deletions, only ΔE56 maintained the reading frame and was predicted to generate an 85 kDa protein. Exons 5‐6 encode an uncharacterized portion of the cytoplasmic N‐terminal domain of a3, a domain not involved in proton translocation. To investigate the effect of ΔE56 on V‐ATPase function, we transformed yeast with plasmids carrying full‐length or truncated Vph1p, the yeast ortholog of a3. Both proteins were expressed; however, ΔE56‐Vph1p transformed yeast failed to grow on Zn2+‐containing plates, a growth assay dependent on V‐ATPase‐mediated vacuolar acidification. In conclusion, our results show that the ΔE56 truncated protein is not functional, suggesting that the mild ARO phenotype observed in the patient is likely due to the residual full‐length protein expression.
B cell epitopes in the intrinsically disordered regions of neuraminidase and hemagglutinin proteins of H5N1 and H9N2 avian influenza viruses for peptide‐based vaccine development
Authors: Ramamurthy Mageshbabu , Sankar Sathish , Abraham Asha Mary , Nandagopal Balaji , Sridharan Gopalan 
Published: May 2019

AbstractAvian influenza viruses (AIV) are very active in several parts of the globe and are the cause of huge economic loss for the poultry industry and also human fatalities. Three dimensional modeling was carried out for neuraminidase (NA) and hemagglutinin (HA) proteins of AIV. The C‐score, estimated TM‐Score, and estimated root‐mean‐square deviation (RMSD) score for NA of H5N1 were −1.18, 0.57 ± 0.15, and 9.8 ± 7.6, respectively. The C‐score, estimated TM‐Score, and estimated RMSD score for NA of H9N2 were −1.43, 0.54 ± 0.15, and 10.5 ± 4.6, respectively. The C‐score, estimated TM‐Score, and estimated RMSD score for HA of H5N1 were −0.03, 0.71 ± 0.12, and 7.7 ± 4.3, respectively. The C‐score, estimated TM‐Score, and estimated RMSD score for HA of H9N2 were −0.57, 0.64 ± 0.13, and 8.9 ± 4.6, respectively.Intrinsically disordered regions were identified for the NA and HA proteins of H5N1 and H9N2 with the use of PONDR program. Linear B cell epitope was predicted using BepiPred 2 program for NA and HA of H5N1 and H9N2 avian influenza strains. Discontinuous epitopes were predicted by Discotope 2 program. The linear epitopes that were considered likely to be immunogenic and within the intrinsically disordered region for the NA of H5N1 was TKSTNSRSGFEMIWDPNGWTGTDSSFSVK, and for H9N2 it was VGDTPRNDDSSSSSNCRDPNNERGAP. In the case of HA of H5N1, it was QRLVPKIATRSKVNGQSG and ATGLRNSPQRERRRKK; for H9N2 it was INRTFKPLIGPRPLVNGLQG and SLKLAVGLRNVPARSSR. The discontinuous epitopes of NA of H5N1 and H9N2 were identified at various regions of the protein structure spanning from amino acid residue positions 90 to 449 and 107 to 469, respectively. Similarly, the discontinuous epitopes of HA of H5N1 and H9N2 were identified in the amino acid residue positions 27 to 517 and 136 to 521, respectively.This study has identified potential and highly immunogenic linear and conformational B‐cell epitopes towards developing a vaccine against AIV both for human and poultry use.
The effect of combined miR‐200c replacement and cisplatin on apoptosis induction and inhibition of gastric cancer cell line migration
Authors: Ghasabi Mehri , Majidi Jafar , Mansoori Behzad , Mohammadi Ali , Shomali Navid , Shirafkan Naghmeh , Baghbani Elham , Kazemi Tohid , Baradaran Behzad 
Published: May 2019

AbstractOne of the major obstacles in the treatment of cancer is resistance to standard chemotherapeutic drugs. According to the numerous reports, miR‐200c is involved in many cancers, especially gastric cancer, and also miR‐200c has been known as an effective factor in the elimination of chemotherapy resistance. The purpose of this study was to explore the potential function and mechanism of miR‐200c and cisplatin in the inhibition of migration and induction of apoptosis in gastric cancer cells. In this study, first, miR‐200c mimics and LNA‐anti‐miR‐200c were transfected into KATOIII cells. Moreover, 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay results revealed that increased miR‐200c expression and cisplatin can more inhibited the proliferation of KATOIII cells. MiR‐200c overexpression inhibited the movement of KATOIII cells in wound healing assay. Subsequently, miR‐200c/cisplatin could suppress colony formation in KATOIII cells. To identify a potential miR‐200c target, we investigated the effect of miR‐200c modulation on RhoE, PTEN, VEGFR, and MMP9 expression levels. Increased miR‐200c expression caused a reduction in VEGFR and MMP9 mRNA and protein, suggesting that VEGFR and MMP9 are targets of miR‐200c. In addition, reverse‐transcription polymerase chain reaction assays showed that RhoE is target gene of miR‐200c and LNA‐anti‐miR‐200c suppressed the expression of PTEN. Flow cytometry and 4′,6‐diamidino‐2‐phenylindole staining analysis indicated that miR‐200c increased the cisplatin‐induced apoptosis which may be associated with suppression of RhoE expression in KATOIII cells, also cell‐cycle analysis showed the arrest of cell‐cycle progression at the G2 phase. These data demonstrated that miR‐200c functioned as a suppressor gene in KATOIII cells. Also, miR‐200c can be a potential therapeutic approach to overcome chemoresistance during cisplatin chemotherapy.
Enrichment of cancer stem‐like cells by the induction of epithelial‐mesenchymal transition using lentiviral vector carrying E‐cadherin shRNA in HT29 cell line
Authors: Saltanatpour Zohreh , Johari Behrooz , Alizadeh Akram , Lotfinia Majid , Majidzadeh‐A Keivan , Nikbin Behrooz , Kadivar Mehdi 
Published: May 2019

AbstractA better understanding of cancer stem cells (CSCs) may facilitate the prevention and treatment of cancers. Epithelial‐mesenchymal transition (EMT) is a process activated during invasion and metastasis of tumors. EMT induction in normal and tumor cells makes them more resistant to chemotherapy. E‐cadherin is a membrane protein and plays a role in tumor invasion, metastasis, and prognosis. Downregulation of E‐cadherin is a hallmark of EMT. Here, we created a model of cancer stem‐like cells enrichment via EMT induction using E‐cadherin downregulation in HT29 cell line using a lentiviral vector carrying shRNA. We aimed to evaluate cancer and anti‐CSC chemotherapeutics screening. The markers of EMT and CSCs were assessed and compared with control cells using flow cytometry, real‐time PCR, immunocytochemistry, western blot, migration assay, invasion assay, and colony formation assay. The transduced cells showed a mesenchymal morphology. High levels of EMT‐related proteins were also expressed. These results confirmed that the transduced cells underwent EMT. In addition, we observed an increased population of E‐cadherin‐downregulated HT29 cell line among the cells expressing colon CSC markers (CD133+ and CD44+) after EMT induction. E‐cadherin‐downregulated cells were morphologically like mesenchymal cells, and the number of CD133+‐ and CD44+‐cells (CSC‐like cells) increased. These cells can be used as stable models to study cancer cells and screening of antitumor therapeutics.
Factors responsible for estivation in spotted stem borer, Chilo partellus (Swinhoe)
Authors: Dhillon Mukesh K. , Hasan Fazil , Tanwar Aditya K. , Bhadauriya Amarpal S. 
Published: May 2019

AbstractDiapause is an important adaptive trait in certain groups of insects at a particular stage, which contribute in controlling insect ecology, phenology, and physiology. Considering the importance of various factors in appropriate combinations, we studied effects of different temperature and photoperiod conditions on larval morphometrics, development, induction, and termination of estivation, and overall mortality due to estivation in Chilo partellus (Swinhoe). Weight, length, and head capsule width of C. partellus larvae significantly increased with increase in temperature and photoperiod treatments upto certain age and duration of exposure. According to Dyar's ratio, overall head capsule width progressed geometrically with minimum constant ratio of 1.20, and second‐degree polynomial regression equations were best fitted to all the test treatment conditions. The mortality of diapausing C. partellus individuals varied between 7.0% and 37.3% under different constant and varying temperature and photoperiod regimes, which at 38°C + 14L:10D reached upto 100.0%. However, the exposure to 30°C + 12.5L:11.5D and 32°C + 13L:11D along with dry food accelerated the process of estivation, wherein the diapause larval survival was significantly higher and overall mortality of individuals due to diapause was lower suggesting these conditions appropriate for induction of estivation in C. partellus. These studies have implications for better understanding the bioecology and population regulation system under varying climatic conditions and devising novel strategies for management of C. partellus.
Low catalytic activity is insufficient to induce disease pathology in triosephosphate isomerase deficiency
Authors: Segal Joanna , Mülleder Michael , Krüger Antje , Adler Thure , Scholze‐Wittler Manuela , Becker Lore , Calzada‐Wack Julia , Garrett Lillian , Hölter Sabine M. , Rathkolb Birgit , Rozman Jan , Racz Ildiko , Fischer Ralf , Busch Dirk H. , Neff Frauke , Klingenspor Martin , Klopstock Thomas , Grüning Nana‐Maria , Michel Steve , Lukaszewska‐McGreal Beata , Voigt Ingo , Hartmann Ludger , Timmermann Bernd , Lehrach Hans , Wolf Eckhard , Wurst Wolfgang , Gailus‐Durner Valérie , Fuchs Helmut , H. de Angelis Martin , Schrewe Heinrich , Yuneva Mariia , Ralser Markus 
Published: May 2019

AbstractTriosephosphate isomerase (TPI) deficiency is a fatal genetic disorder characterized by hemolytic anemia and neurological dysfunction. Although the enzyme defect in TPI was discovered in the 1960s, the exact etiology of the disease is still debated. Some aspects indicate the disease could be caused by insufficient enzyme activity, whereas other observations indicate it could be a protein misfolding disease with tissue‐specific differences in TPI activity. We generated a mouse model in which exchange of a conserved catalytic amino acid residue (isoleucine to valine, Ile170Val) reduces TPI specific activity without affecting the stability of the protein dimer. TPIIle170Val/Ile170Val mice exhibit an approximately 85% reduction in TPI activity consistently across all examined tissues, which is a stronger average, but more consistent, activity decline than observed in patients or symptomatic mouse models that carry structural defect mutant alleles. While monitoring protein expression levels revealed no evidence for protein instability, metabolite quantification indicated that glycolysis is affected by the active site mutation. TPIIle170Val/Ile170Val mice develop normally and show none of the disease symptoms associated with TPI deficiency. Therefore, without the stability defect that affects TPI activity in a tissue‐specific manner, a strong decline in TPI catalytic activity is not sufficient to explain the pathological onset of TPI deficiency.