Journal of Cell and Molecular Research最新文献

{"title":"Identification of Suitable Housekeeping Genes for Quantitative Gene Expression Analysis During Retinoic Acid-induced Differentiation of Embryonal Carcinoma NCCIT Cells","authors":"S. Soltanian, Mahboubeh Sheikhbahaei","doi":"10.22067/JCMR.V12I2.88457","DOIUrl":"https://doi.org/10.22067/JCMR.V12I2.88457","url":null,"abstract":"Real-time quantitative PCR (qRT-PCR) is often used as an effective experimental method for analyzing gene expression. In this method, normalization of target gene expression levels must be performed using housekeeping genes (HKGs). HKGs are used to compensate for difference between samples due to diverse quality and quality of RNAs and different reverse transcription yield. For an ideal reference gene, constant expression levels across different samples of one experiment is necessary. In the current study, expression stability of four candidate references genes including Beta actin (ACTB), glyceraldeyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine guanine phosphoribosyl transferase (HPRT1) and Beta-2-Microglobulin (β2M) following retinoic acid (RA) treatment in embryonal carcinoma NCCIT cells were evaluated.NCCIT cells were exposed to RA (10 µM) for 14 days to induce differentiation. RT-qPCR for candidate references genes was performed and normalization between untreated and RA-treated cells was performed using identical sample input amounts. Expression of OCT4, SOX2, NANOG during RA-induced differentiation was assessed by quantitative real-time PCR. RT-qPCR results indicated significant difference in expression level of GAPDH between untreated (Ct mean: 19.36667± 0.28) and RA-treated (Ct mean: 28.94± 0.18) NCCIT cells. However, transcriptional level of ACTB, HPRT and β2M remained unchanged after RA treatment. qRT-PCR analysis using ACTB, HPRT and β2M showed treatment of NCCIT cells with RA lead to significant down regulation of OCT4 (79%), NANOG (71%) and SOX2 (96%) transcript. ACTB, HPRT and β2M were recognized as valid reference genes for analysis of gene expression during RA-induced differentiation of NCCIT cells, while GAPDH was not suitable.","PeriodicalId":30607,"journal":{"name":"Journal of Cell and Molecular Research","volume":"12 1","pages":"104-110"},"PeriodicalIF":0.0,"publicationDate":"2021-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49636443","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative in silico Survey of Retinoic Acids as Putative Biomolecules Against VEGF Receptors: A Glycomics-based Approach","authors":"A. Fallah, Esmaeil Samadian, Azadeh Mohammad-Hasani, A. Colagar","doi":"10.22067/JCMR.V12I2.89670","DOIUrl":"https://doi.org/10.22067/JCMR.V12I2.89670","url":null,"abstract":"In this study, putative interactions between all of the retinoic acid (RA) ligands (all-trans (At), 9-cis (9c), and 13-cis (13c)), and VEGF receptors (VEGFR-1, -2 and -3) were investigated. It was performed considering the glycosylation status of the receptors to achieve a more reliable mode of interactions based on glycomics. We found that RAs may have a higher affinity for ligand-binding domains in VEGFRs. Furthermore, all RA isomers can strongly attach to VEGFR-3 receptor in comparison to other ones. It was also demonstrated that receptor dimerization of RAs may be less targeted. Moreover, regarding post-translational modifications, glycosylated structures showed conflicting binding energies. RAs may target the human vasculature, specifically lymph vessels, through VEGFR-3. In addition, the ligand binding-mediated activation of VEGFRs may be affected by these agents. Also, the glycosylation status of the receptors can interfere with these manners. Furthermore, our results confirmed that the consideration of carbohydrates in crystal structures is essential for a better interpretation of ligand/receptor interactions during drug discovery studies. Even though these observations improved our understanding of the binding patterns of RAs to VEGFRs, validation of these results needs further analysis to introduce these biomolecules as anti-VEGF remedies.","PeriodicalId":30607,"journal":{"name":"Journal of Cell and Molecular Research","volume":"12 1","pages":"87-96"},"PeriodicalIF":0.0,"publicationDate":"2021-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46014289","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of Bacterial Proteins in the Gut of Sunn pest, Eurygaster integriceps","authors":"M. Saadati, M. Toorchi","doi":"10.22067/JCMR.2021.64305.0","DOIUrl":"https://doi.org/10.22067/JCMR.2021.64305.0","url":null,"abstract":"Proteomics is a powerful tool to identify effective proteins in the biochemical reactions of the insect body. Many proteins were reported in the gut lumen and tissues which are essential to complete the physiological role of the alimentary canal of Sunn pest. The gut microbiome of insects has a key role in the digestive process. In this study, for the first time, gut proteins of adult Sunn pest were extracted. These proteins were visualized and identified with two-dimensional polyacrylamide gel electrophoresis and mass spectrometry, respectively. Newly identified proteins include pyruvate dehydrogenase, oxidoreductase FAD-binding protein, hypothetical proteins, glycerol-3 phosphate dehydrogenase, conserved hypothetical proteins, ABC excinucleases, ABC-type transport systems and molecular chaperones. The accumulation of these metabolism proteins in the gut of Sunn pest indicates the importance of symbiotic proteins in the improvement of digestive activities and insect-bacteria interactions. Results suggest that the identified bacterial proteins can be considered as effective proteins in the process of nutrition and provide more gut-derived targets for enzyme engineering and development of biopesticides.","PeriodicalId":30607,"journal":{"name":"Journal of Cell and Molecular Research","volume":"12 1","pages":"97-103"},"PeriodicalIF":0.0,"publicationDate":"2021-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49387126","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Signal Transduction of Unique RAS Family Member towards Cell Survival","authors":"Saeideh Nakhaei-Rad","doi":"10.22067/JCMR.V12I2.2102.1002","DOIUrl":"https://doi.org/10.22067/JCMR.V12I2.2102.1002","url":null,"abstract":"Small GTPases of RAS act as central regulators of intracellular signal transduction and translate external stimuli to the various cellular responses. Embryonic stem cell expressed RAS (ERAS) is a member of the RAS family that is specifically expressed in undifferentiated mouse embryonic stem cells, hepatic stellate cells and diverse human tumors, such as gastric, breast, brain, pancreatic, and colorectal tumors. Although ERAS belongs to GTPase family, it is an inefficient enzyme to hydrolyze GTP to GDP. Therefore, it remains mainly in its GTP-bound active form and contributes to sustained signal transduction. In comparison with classical members (HRAS, NRAS and KRAS4B), ERAS is known as a unique member, due to its temporal expression, remarkable amino acid sequence deviations and functional differences. Notably, ERAS has been recently proposed as a potential marker for drug resistance in several human tumors. In this minireview, I compare in great detail the biochemical properties of ERAS with conventional members of the RAS family, and discuss the main ERAS function in the control of the PI3K-AKT-mTORC survival pathway. Targeting this pathway may sensitize ERAS expressing cell populations to chemotherapy.","PeriodicalId":30607,"journal":{"name":"Journal of Cell and Molecular Research","volume":"12 1","pages":"65-73"},"PeriodicalIF":0.0,"publicationDate":"2021-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47252695","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cloning and Expression of a Fusion Protein Containing Highly Epitopic Regions of Clostridium perfringens Epsilon Toxin and Clostridium novyi Alpha Toxin","authors":"M. Mehrvarz, M. Najafi, T. Z. Salehi, B. Majidi","doi":"10.22067/JCMR.2021.68924.1003","DOIUrl":"https://doi.org/10.22067/JCMR.2021.68924.1003","url":null,"abstract":"Clostridium perfringens and novyi species are two important toxin-producing pathogens which pose a risk to the livestock health. Epsilon and alpha toxins are major toxins of these two pathogens, respectively. Advances in current vaccine industrialization lead to the utilization of toxin epitopes instead of the whole pathogen/toxoids to produce novel vaccines. In the present study, bioinformatics approaches were applied to design a fused protein containing both toxin fragments of interest with the highest antigenicity score for B-cells. To do so different specialized algorithms including I-TASSER, IEDB, ElliPro, PyDock and CLC Main Workbench were applied. The chimeric protein was successfully cloned, expressed, and purified using an immobilized-metal affinity chromatography for His-tagged proteins. During in vivo experiments on rabbits, the levels of immunization provided by the recombinant protein or native alpha and epsilon toxins were compared based on serological studies. Results indicated that the designed protein was able to stimulate effective immune responses against both alpha and epsilon toxins. This can be used as a proper strategy to design novel peptide-based subunit vaccines. Clostridium perfringens and novyi species are two important toxin-producing pathogens which pose a risk to the livestock health. Epsilon and alpha toxins are major toxins of these two pathogens, respectively. Advances in current vaccine industrialization lead to the utilization of toxin epitopes instead of the whole pathogen/toxoids to produce novel vaccines. In the present study, bioinformatics approaches were applied to design a fused protein containing both toxin fragments of interest with the highest antigenicity score for B-cells. To do so different specialized algorithms including I-TASSER, IEDB, ElliPro, PyDock and CLC Main Workbench were applied. The chimeric protein was successfully cloned, expressed, and purified using an immobilized-metal affinity chromatography for His-tagged proteins. During in vivo experiments on rabbits, the levels of immunization provided by the recombinant protein or native alpha and epsilon toxins were compared based on serological studies. Results indicated that the designed protein was able to stimulate effective immune responses against both alpha and epsilon toxins. This can be used as a proper strategy to design novel peptide-based subunit vaccines.","PeriodicalId":30607,"journal":{"name":"Journal of Cell and Molecular Research","volume":"12 1","pages":"56-64"},"PeriodicalIF":0.0,"publicationDate":"2021-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48346973","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification and Functional Profiling of Differentially Expressed Extracellular Vesicle-free MicroRNAs for Efficient Prostate Cancer Diagnosis","authors":"Mina Jahandideh, Ebrahim Barzegari","doi":"10.22067/JCMR.V12I1.87155","DOIUrl":"https://doi.org/10.22067/JCMR.V12I1.87155","url":null,"abstract":"MicroRNAs are interesting as cancer diagnostic and prognostic biomarkers because of their unique tissue expression profiles, higher stability in the blood in comparison to mRNAs, and the possibility for reliable quantification. In the case of prostate cancer (PCa), it is currently emphasized to explore new biomarkers, particularly from microRNAs which are freely available in the bloodstream. In this study, the gene expression omnibus database (GEO), a repository of microarray data for PCa circulating extracellular vesicle-free microRNAs profiling, was analyzed for differentially expressed miRNAs (DE-miRs). Top 20 most differentially expressed miRs with significant (adjusted p-value < 0.01) high expression (fold change) levels were extracted by the simultaneous application of different filtering criteria. Then, microRNA-gene networks were constructed for the two sets of positively (n=20) or negatively (n=20) regulated miRNAs. Gene ontology annotations of the target gene sets were also extracted and analyzed. Results indicated that human miR-1587, miR-223-3p, miR-3125, and miR-642b-3p are highly significant DE-miRs in PCa. In addition, human miR-4459, miR-1273g, miR 642a-3p, and miR-642b-3p were identified as top-ranked hubs in the relevant miRNA-gene networks. FOXK1, PML, CD24, ATN1, BAZ2A, CDKN1A, NUFIP2, and HARNPU were identified as microRNA target genes with significant dysregulation. miR-4459, miR-1273g-3p, miR-3135b, miR-5001-5p, and miR-1587 were proposed as novel microRNAs with the potential to be utilized as diagnostic biomarkers of prostate cancer among circulating vesicle-free miRNAs.","PeriodicalId":30607,"journal":{"name":"Journal of Cell and Molecular Research","volume":"12 1","pages":"10-21"},"PeriodicalIF":0.0,"publicationDate":"2020-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47552102","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Element Accumulations in Liver and Kidney Tissues of Some Bony Fish Species in the Southwest Caspian Sea","authors":"M. Sattari, M. Bibak, S. Bakhshalizadeh, Mohammad Forouhar Vajargah","doi":"10.22067/JCMR.V12I1.85975","DOIUrl":"https://doi.org/10.22067/JCMR.V12I1.85975","url":null,"abstract":"The Caspian Sea is the largest inland body of water in the world and so has both common characteristics of seas and lakes with over 153 fish species which inhabit the sea and its basin. However, little is known about the trace element (TE) contaminations (TECs) in its tissues. In the present study, 122 specimens of three fish species including Rutilus caspius (Roach, n=71), Leuciscus aspius (Asp, n=20), and Tinca tinca (Tench, n=31) were collected from three different fisheries regions (i.e. Astara, Anzali and Kiashahr) of the southern part of the Caspian Sea from September 2017 to June 2018. Inductively coupled plasma optical emission spectrometry (ICP-OES) was employed to measure TE levels in different fish tissues. An attempt was made to assess possible influences of habitat on element accumulation in the liver and kidney of three fish species in the southwest of the Caspian Sea basin. Some elements including Ca, K, Mg, P, S, Sc, and Sr showed different concentrations in the liver and kidney. Also their levels were significantly different between freshwater resident (Tench) and marine (Roach) species (p < 0.05). The differences among TECs in the liver and kidney of Roach, Asp and Tench were reduced to three components using principal component analysis (PCA). Results indicated that 83.60% of the total variability is related to TEs such as Cu, Fe, Sr, Ca, S, Na, Mg, K, and Al. The impact of habitat variability on the element accumulation was confirmed through linear chart obtained for liver and kidney (as body filtering organs) of Roach and Asp as marine residents as well as Tench as a freshwater resident. This could illustrate the borderline created by these habitats.","PeriodicalId":30607,"journal":{"name":"Journal of Cell and Molecular Research","volume":"12 1","pages":"33-40"},"PeriodicalIF":0.0,"publicationDate":"2020-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46137004","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Network Analysis of Differential Gene Expression to Identify Hub Genes in Ovarian Cancer","authors":"A. Siavoshi, Mahdieh Taghizadeh, E. Dookhe, M. Piran, Mahsa Saliani, Shahla Mohammad, Ganji","doi":"10.22067/JCMR.V12I1.85654","DOIUrl":"https://doi.org/10.22067/JCMR.V12I1.85654","url":null,"abstract":"Epithelial ovarian cancer (EOC), as a challenging disease among women with poor prognosis and unclear molecular pathogenesis, each year is responsible for 140000 deaths globally. Recent progress in the field revealed the importance of proteins as key players of different biological events. Considering the complicated protein interactions, taking a deeper look at protein-protein interactions (PPIs) could be considered as a superior strategy to unravel complex mechanisms encountered with regulatory cell signaling pathways of ovarian cancer. Hence, PPI network analysis was performed on differentially expressed genes (DEGs) of ovarian cancer to discover hub genes which have the potential to be introduced as biomarkers with clinical utility. A PPI network with 600 DEGs was constructed. Network topology analysis determined UBC, FN1, SPP1, ACTB, GAPDH, JUN, and RPL13A, with the highest Degree (K) and betweenness centrality (BC), as shortcuts of the network. KEGG pathway analysis showed that these genes are commonly enriched in ribosome and ECM-receptor interaction pathways. These pivotal hub genes, mainly UBC, FN1, RPL13A, SPP1, and JUN have been reported previously as potential prognostic biomarkers of different types of cancer. However, further experimental molecular studies and computational processes are required to confirm the function and association of the identified hub genes with epithelial ovarian cancer prognosis.","PeriodicalId":30607,"journal":{"name":"Journal of Cell and Molecular Research","volume":"12 1","pages":"1-9"},"PeriodicalIF":0.0,"publicationDate":"2020-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45712593","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Association of TOMM40, CHAT and SORL1 Polymorphisms with the Alzheimer’s Disease in the Turkish-speaking Azeri population in Northwest of Iran","authors":"Nazanin Rohani, Mohammad Khalaj-Kondari, Majid Khodayi-Shahrak, M. Talebi, M. feizi","doi":"10.22067/JCMR.V12I1.87325","DOIUrl":"https://doi.org/10.22067/JCMR.V12I1.87325","url":null,"abstract":"Recent genome-wide association studies have introduced several genetic variants which contribute to the late-onset Alzheimer's disease (LOAD). Polymorphisms of CHAT, TOMM40, and SORL1 genes have been reported to be associated with the LOAD phenotype. This study was endeavored to evaluate the association of the CHAT rs3810950, TOMM40 rs1160985 and SORL1 rs11218304 polymorphisms with the LOAD in the Turkish-speaking  Azeri population of northwest Iran. In a case-control study, we included 174 cases: 88 cases with LOAD diagnosis and 86 healthy individuals. Peripheral blood samples were collected and the genomic DNA of all participants were extracted. Genotyping was carried out by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. We did not observe any significant association between the CHAT rs3810950 and SORL1 rs11218304 alleles with the LOAD. However, both the TOMM40 rs1160985 minor allele T and TT genotype showed significant negative associations with the LOAD. Hence, the TOMM40 rs1160985 polymorphism could be considered as a protective genetic factor against the LOAD in the Turkish-speaking Azeri population of northwest Iran.","PeriodicalId":30607,"journal":{"name":"Journal of Cell and Molecular Research","volume":"12 1","pages":"41-48"},"PeriodicalIF":0.0,"publicationDate":"2020-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44501168","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of Breast Cancer Associated Putative Functional Single Nucleotide Polymorphisms in the Iranian Population through In Silico Analyses","authors":"Vida Nadafi Sichani, S. Emami, Morteza Bitaraf Sani, M. Nassiri, V. Gopalan","doi":"10.22067/JCMR.V12I1.87629","DOIUrl":"https://doi.org/10.22067/JCMR.V12I1.87629","url":null,"abstract":"Previous studies have found several distinct alleles at both levels of transcriptional activity and protein-DNA binding manners in breast cancer patients vs. healthy individuals through multi-step experimental approaches. This study presents a computational-based model to investigate the regulatory potential and functional properties of disease-related non-coding single nucleotide polymorphisms (SNPs) variants through several online in silico tools in the Iranian population. The association between the risk of breast cancer and its putative single nucleotide polymorphisms in the Iranian population was investigated through SNPedia database and genome-wide association studies (GWAS). Furthermore, a meta-analysis was performed by Comprehensive Meta-Analysis (CMA) software. Functional analyses were carried out through LDlink, HaploReg, and RegulomeDB. The impact of each single nucleotide polymorphism on gene expression profiles and transcription factor binding sites were predicted by the RegulomeDB. \"5\", \"6\", and \"1d\" scores were assigned to rs3746444, rs1062577, and rs1049174 by this scoring system, respectively. RegulomeDB scores of rs3746444-MYH7B/MIR499A and rs1062577-ESR1 suggested that they are not putative functional single nucleotide polymorphisms; and may not associate with significant eQTL signals. The “1d” score for rs1049174-RP11-277P12.20 confirmed an association with the expression of the target gene. Proxy variants rs6088678 and rs2617160 have been identified using LDlink in non-coding segments. They were in strong linkage disequilibrium (LD) with single nucleotide polymorphisms rs3746444 and rs1049174, respectively. Also, non-coding variants rs6088678-TRPC4AP and rs2617160- RP11-277P12.20 with high-ranked scores showed the strongest related-expression. This work provides a rapid and direct in silico-based approach for the identification of functional genetic variants in the breast cancer. These analyses were conducted to evaluate the association of intended SNPs with the regulatory elements of histones, DNases, motif changes, and selected eQTL signals. It can be extended to some other complex single nucleotide polymorphism-related diseases.","PeriodicalId":30607,"journal":{"name":"Journal of Cell and Molecular Research","volume":"12 1","pages":"22-32"},"PeriodicalIF":0.0,"publicationDate":"2020-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47472753","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}