I. Kocherova, Rut Bryl, I. Crha, P. Ventruba, J. Žáková, M. Ješeta
{"title":"The extracellular reactive oxygen species levels in primary in vitro culture of human ovarian granulosa and cumulus cells","authors":"I. Kocherova, Rut Bryl, I. Crha, P. Ventruba, J. Žáková, M. Ješeta","doi":"10.2478/acb-2020-0014","DOIUrl":"https://doi.org/10.2478/acb-2020-0014","url":null,"abstract":"Abstract In the female reproductive tract, reactive oxygen species (ROS) may exert physiological and pathophysio-logical effects. Although ROS play an essential role as the signaling molecules, their excessive accumulation contributes to the pathogenesis of many reproductive processes. In the ovarian follicle, ROS affect multiple physiological processes, including oocyte maturation and fertilization. However, a lack of studies showing to which extend ovarian granulosa and cumulus cells can contribute to the development of oxidative stress within the ovarian follicle. In the presented research, the extracellular ROS accumulation level was investigated using GCs and CCs primary in vitro cultures. The obtained results demonstrated a steady decrease in extracellular ROS level during GCs primary culture. By contrast, ROS concentration in CCs conditioned medium increased gradually between the first and the seventh days of culture. The observed changes may reflect the proliferation status and metabolic activity of GCs and CCs during in vitro culture. Additionally, the elevated ROS level at respective points of time could occur as a consequence of culture in atmospheric oxygen. The distinct function and localization within the ovarian follicle may explain the differences between GCs and CCs oxygen metabolism. Running title: Reactive oxygen species in primary culture of human follicular cells","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"8 1","pages":"112 - 117"},"PeriodicalIF":0.0,"publicationDate":"2020-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44512684","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}
K. Stefańska, Rut Bryl, Greg Hutchings, J. Shibli, M. Dyszkiewicz-Konwińska
{"title":"Human umbilical cord stem cells – the discovery, history and possible application","authors":"K. Stefańska, Rut Bryl, Greg Hutchings, J. Shibli, M. Dyszkiewicz-Konwińska","doi":"10.2478/acb-2020-0009","DOIUrl":"https://doi.org/10.2478/acb-2020-0009","url":null,"abstract":"Abstract Stem cells are now one of the most extensively studied fields in experimental medicine. Although embryonic stem cells (ESCs) have the capability to differentiate into lineages derived from all three primary germ layers, their use remains ethically controversial. Mesenchymal stromal cells’ (MSCs) utilization is not burdened with such concerns. MSCs were primarily characterized as fibroblastic cells from bone marrow stroma, with the ability to adhere to plastic and form clonogenic colonies (CFU-Fs – colony-forming unit-fibroblasts). Subsequent studies led to their isolation from various other adult or perinatal tissues, including the umbilical cord. Although in most cases MSCs are obtained from umbilical cords’ stroma (Wharton’s jelly), it has been demonstrated that both cord’s lining and endothelial layer of umbilical vein possess the cells that fulfill the minimal criteria for MSCs published in 2006 by the International Society for Cellular Therapy. Since the initial discovery, a number of pre-clinical and clinical studies were conducted with the use of MSCs derived from umbilical cords, targeting diseases such as cancer or autoimmune diseases. Moreover, their secretome may be even more beneficial, providing an opportunity to create cell-free therapies. Running title: The history of human umbilical cord stem cells","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"8 1","pages":"78 - 82"},"PeriodicalIF":0.0,"publicationDate":"2020-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42880911","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}
M. Ješeta, M. Mysková, J. Žáková, I. Crha, K. Crha, E. Chmelikova, E. Kistanova, P. Ventruba
{"title":"Can oocytes repair fragmented DNA of spermatozoa?","authors":"M. Ješeta, M. Mysková, J. Žáková, I. Crha, K. Crha, E. Chmelikova, E. Kistanova, P. Ventruba","doi":"10.2478/acb-2020-0008","DOIUrl":"https://doi.org/10.2478/acb-2020-0008","url":null,"abstract":"Abstract Approximately half of the cases of infertility are due to male factor. In many cases the underlying cause of male infertility is not discovered and, therefore, the condition is considered idiopathic. Examination of morphology, motility, concentration and total sperm count is very important but not sufficient for complex men fertility evaluation. Sperm DNA integrity is a very important one. Sperm DNA can be fragmented by several internal or external factors. In immature sperm cells, the DNA can be repaired by reparatory mechanisms of spermatogonia or spermatocytes. However, in a haploid mature sperm cell, these fragments can not be repaired by male and the fragmented DNA is transferred to oocyte. Oocytes are able to repair male fragmented DNA after their fertilization. A quality embryo can repair damaged sperm DNA and the repair system depends on cytoplasmic and genomic quality of the oocyte. The ability of oocyte to repair sperm DNA strong depend on quality of fertilized oocytes. Running title: Oocyte and DNA repair","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"8 1","pages":"73 - 77"},"PeriodicalIF":0.0,"publicationDate":"2020-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46398583","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}
K. Stefańska, Rut Bryl, Lisa Moncrieff, Nelson Pinto, J. Shibli, M. Dyszkiewicz-Konwińska
{"title":"Mesenchymal stem cells – a historical overview","authors":"K. Stefańska, Rut Bryl, Lisa Moncrieff, Nelson Pinto, J. Shibli, M. Dyszkiewicz-Konwińska","doi":"10.2478/acb-2020-0010","DOIUrl":"https://doi.org/10.2478/acb-2020-0010","url":null,"abstract":"Abstract Mesenchymal stem cells are currently one of the most extensively studied topics in experimental medicine, given their unique properties in terms of immunomodulation, multiple factors secretion and homing to injured tissue sites. Such characteristics were proven to be invaluable in various disease management treatments, for example in cancer, tissue regeneration or immunologic/inflammatory-related disorders. MSCs were first isolated from bone marrow in 1960-1970’s and were characterized as cells with fibroblastoid shape and osteogenic potential, which form clonogenic colonies (CFU-F – colony-forming unit-fibroblast). Nowadays the term ‘mesenchymal stem cells’ is used in regards to all of the cells meeting minimal criteria published in 2006 by the International Society for Cellular Therapy, however the name ‘mesenchymal stromal cells’ has been suggested to be more appropriate. Regardless of the name controversy, these cells exhibit multilineage differentiation potential, self-renewal ability, adhere to plastic and express specific surface antigens. In 2011 the first commercial product based on MSCs was developed and many more are expected to emerge. This review focuses on a historical perspective concerning studies on MSCs, controversies regarding their name and their characteristics and clinical utilization. Running title: The history of mesenchymal stem cells","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"8 1","pages":"83 - 87"},"PeriodicalIF":0.0,"publicationDate":"2020-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48458182","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}
B. Borowiec, Greg Hutchings, Rut Bryl, M. A. Melo, J. Shibli, M. Dyszkiewicz-Konwińska
{"title":"The advances in human oral biology and biotechnology","authors":"B. Borowiec, Greg Hutchings, Rut Bryl, M. A. Melo, J. Shibli, M. Dyszkiewicz-Konwińska","doi":"10.2478/acb-2020-0011","DOIUrl":"https://doi.org/10.2478/acb-2020-0011","url":null,"abstract":"Abstract The oral cavity is a very specific and complicated structure. It plays the role of a speech apparatus but is also a gate and a preliminary processing plant for fuel nourishing the body. Analyzing the embryological development of the oral cavity, we can distinguish here the stages of growth, migration and fusion. This processes mainly involves the formation of branched arches that appear between the fourth and fifth weeks of fetal development. In recent years, interest in researching its various features has been still growing. After a thorough description and anatomical examination, researchers began to think about its other possibilities than just crushing, throwing, swallowing, bringing out the sounds and articulating words. It turns out that the whole range of its potential is not limited to just a few functions mentioned above. The following review article covers specific studies that have emerged most frequently over the past year. You will find here information about the microbiome of the oral cavity which plays an significant role in maintaining a proper physiological environment of the oral cavity, oral tumors and how the SCC or squamous cell carcinoma is histologically the most common and its treatment, even in the context of the Covid-19 pandemic, as well as the least discovered and still fascinating for scientists wound healing processes and unknown exosomes that can transport biological molecules in their original, intact and active form, including nucleic acids and proteins with their signaling pathways. Running title: The advances in human oral biology and biotechnology","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"8 1","pages":"88 - 94"},"PeriodicalIF":0.0,"publicationDate":"2020-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43891736","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}
M. Nawrocki, S. Kałużna, K. Jopek, Greg Hutchings, B. Perek, M. Jemielity, A. Malińska, B. Kempisty, P. Mozdziak, M. Nowicki
{"title":"Aortocoronary conduits may show a different inflammatory response - comparative study at transcript level","authors":"M. Nawrocki, S. Kałużna, K. Jopek, Greg Hutchings, B. Perek, M. Jemielity, A. Malińska, B. Kempisty, P. Mozdziak, M. Nowicki","doi":"10.2478/acb-2020-0003","DOIUrl":"https://doi.org/10.2478/acb-2020-0003","url":null,"abstract":"Abstract Coronary artery bypass grafting (CABG), together with percutaneous coronary intervention (PCI), are both still the most efficient procedures for myocardial revascularization to treat advanced coronary artery disease (CAD). Donor blood vessels used in CABG are usually the internal thoracic artery (ITA) and saphenous vein (SV). The importance of inflammation and inflammatory pathways in graft patency is well established. Nevertheless, not all molecular mechanisms underlying the inflammatory process appear to be clear. Employing the expressive microarray approach to analyze the transcriptome of both venous and arterial grafts, five GO BP terms has been selected: “cellular response to interferon-gamma”, “inflammatory response”, “interferon-gamma-mediated signaling pathway”, “response to interferon-gamma” and “positive regulation of inflammatory response”. This study aimed to evaluate potential molecular factors that could be characteristic markers for both SV and ITA conduits. Running title: Aortocoronary conduits may show a different inflammatory response","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"132 1","pages":"24 - 34"},"PeriodicalIF":0.0,"publicationDate":"2020-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69077820","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}
B. Borowiec, S. Ciesiółka, Krzysztof Janowicz, P. Celichowski, A. Bryja, Lisa Moncrieff, Greg Hutchings, C. Dompe, B. Kempisty, J. Shibli, P. Mozdziak, K. Gerreth, M. Dyszkiewicz-Konwińska
{"title":"The processes of homeostasis, chemotaxis and organic and inorganic response are significantly up-regulated during short-term oral mucosal cells in vitro cultivation","authors":"B. Borowiec, S. Ciesiółka, Krzysztof Janowicz, P. Celichowski, A. Bryja, Lisa Moncrieff, Greg Hutchings, C. Dompe, B. Kempisty, J. Shibli, P. Mozdziak, K. Gerreth, M. Dyszkiewicz-Konwińska","doi":"10.2478/acb-2020-0006","DOIUrl":"https://doi.org/10.2478/acb-2020-0006","url":null,"abstract":"Abstract Mucous membranes appear in various parts of the whole body performing similar functions. However, they differ based on where the mucosa is located. It functions as a barrier in such systems as: respiratory, urogenital and digestive . In this study we will be focusing strictly on the oral mucosa. Keratinocytes and fibroblasts, which mainly form the structure of the oral mucosa, are subjected to numerous factors. Being one of the million parts that build the animal organism, they are involved in various processes. In this study, we will try to confirm that in the in vitro culture of oral mucosa cells, the expression of our selected genes undergoes significant changes which are tied to such processes as: homeostasis, chemotaxis and organic/inorganic response of the organism. For this study, 20 pubertal crossbred Landrace gilts were used. After slaughter, samples of buccal pouch mucosa were obtained and transported to the laboratory. The excised tissue was prepared and processed due to protocols. The final pellet was resuspended in supplemented DMEM. Once the cultures attained 70–80% confluency, they were passaged. Total RNA from each pooled sample was subjected to two rounds of sense cDNA amplification. The cDNA was processed on microarrays. Analysis of the scanned arrays was performed. The files were imported into downstream data analysis software. The DAVID analysis showed that differently expressed genes belongs to 56 Gene ontology groups. In this paper we focused on “cellular divalent inorganic cation homeostasis”, “chemical homeostasis”, “chemotaxis”, “homeostatic process” and “response to organic substance” GO BP terms. These sets of genes were subjected to hierarchical clusterization procedure. In summary, the data we collected showed primarily changes in gene expression that occurred in the thirty-day cell culture of oral mucosa tissue. We assume that indicated genes could be new gene markers for studied processes. Running title: Homeostasis in oral mucosa cells","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"8 1","pages":"50 - 59"},"PeriodicalIF":0.0,"publicationDate":"2020-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46135721","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}
M. Brązert, W. Kranc, K. Jopek, B. Kempisty, L. Pawelczyk
{"title":"New markers of human cumulus oophorus cells cultured in vitro – transcriptomic profile","authors":"M. Brązert, W. Kranc, K. Jopek, B. Kempisty, L. Pawelczyk","doi":"10.2478/acb-2020-0007","DOIUrl":"https://doi.org/10.2478/acb-2020-0007","url":null,"abstract":"Abstract The presence of CCs around the oocyte after ovulation is one of the key elements contributing to oocyte developmental competence. In the presented study, we used CCs from 12 patients aged 18-40 diagnosed with infertility. After harvesting cells on day 1, 7, 15 and 30 of culture, total RNA was isolated and transcriptomic analysis was performed. The DAVID software indicated the following GO BP terms: “cell junction organization”, “cell migration”, “cell morphogenesis involved in differentiation”, “cell morphogenesis” and “cell motility”. Of the genes belonging to all ontological groups, the most downregulated were: SLC7A8, DFNB31, COL1A1, CDC42SE1, TGFBR3, HMGB1, with the most upregulated genes being: ANXA3, KIAA1199, HTR2B, VCAM1, DKK1. While many studies focus on attempts to obtain fully competent oocytes, scientists still have difficulty attaining adequate results in vitro. Lack of adequate knowledge often results in low in vitro fertilization efficiency. Therefore, our research focuses on CCs cells, thanks to which the oocyte most likely acquires developmental competence. The main purpose of the study was to identify the potential molecular markers responsible for cell junction organization, migration, differentiation, morphogenesis and motility. Running title: New markers of human cumulus oophorus cells cultured in vitro","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"8 1","pages":"60 - 72"},"PeriodicalIF":0.0,"publicationDate":"2020-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42721204","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}
Lisa Moncrieff, I. Kocherova, A. Bryja, W. Kranc, Joanna Perek, M. Kulus, M. Ješeta, C. Dompe, Greg Hutchings, Krzysztof Janowicz, P. Celichowski, M. Bruska, M. Zabel, B. Kempisty, P. Mozdziak
{"title":"Transcriptomic profile of genes encoding proteins responsible for regulation of cells differentiation and neurogenesis in vivo and in vitro – an oocyte model approach","authors":"Lisa Moncrieff, I. Kocherova, A. Bryja, W. Kranc, Joanna Perek, M. Kulus, M. Ješeta, C. Dompe, Greg Hutchings, Krzysztof Janowicz, P. Celichowski, M. Bruska, M. Zabel, B. Kempisty, P. Mozdziak","doi":"10.2478/acb-2020-0001","DOIUrl":"https://doi.org/10.2478/acb-2020-0001","url":null,"abstract":"Abstract The growth and development of the oocyte is essential for the ovarian follicle. Cumulus cells (CCs) - a population of granulosa cells - exchange metabolites, proteins and oocyte-derived paracrine factors with the oocyte through gap junctions, to contribute to the competency and health of the oocyte. This bi-directional communication of the cumulus-oocyte complex could be better understood through the micro-analysis of a porcine oocyte gene expression before in vitro maturation (IVM) and after. Additionally, the study of the somatic and gamete cells differentiation capability into neuronal lineage would be promising for future stem cell research as granulosa cells are easily accessible waste material from in vitro fertilization (IVF) procedures. Therefore, in this study, the oocytes of 45 pubertal Landrace gilts were isolated and the protein expression of the COCs were analyzed through micro-analysis techniques. Genes belonging to two ontological groups: neuron differentiation and negative regulation of cell differentiation have been identified which have roles in proliferation, migration and differentiation. Twenty identified porcine oocyte genes (VEGFA, BTG2, MCOLN3, EGR2, TGFBR3, GJA1, FST, CTNNA2, RTN4, MDGA1, KIT, RYK, NOTCH2, RORA, SMAD4, ITGB1, SEMA5A, SMARCA1, WWTR1 and APP) were found to be down-regulated after the transition of IVM compared to in vitro. These results could be applied as gene markers for the proliferation, migration and differentiation occurring in the bi-directional communication between the oocyte and CCs. Running title: Differentiation and neurogenesis in oocyte cells","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"8 1","pages":"1 - 11"},"PeriodicalIF":0.0,"publicationDate":"2020-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47786917","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}
S. Kałużna, M. Nawrocki, K. Jopek, Greg Hutchings, B. Perek, M. Jemielity, B. Kempisty, A. Malińska, P. Mozdziak, M. Nowicki
{"title":"In search of markers useful for evaluation of graft patency - molecular analysis of ‘muscle system process’ for internal thoracic artery and saphenous vein conduits","authors":"S. Kałużna, M. Nawrocki, K. Jopek, Greg Hutchings, B. Perek, M. Jemielity, B. Kempisty, A. Malińska, P. Mozdziak, M. Nowicki","doi":"10.2478/acb-2020-0002","DOIUrl":"https://doi.org/10.2478/acb-2020-0002","url":null,"abstract":"Abstract Coronary artery bypass graft (CABG) is the surgical method most commonly used to treat coronary artery disease (CAD). The vessels that are used in CABG are usually the internal thoracic artery (ITA) and the saphenous vein (SV). Transplant patency is one of the most important factors affecting transplant success. In this study, we used an expressive microarray method, approved by RT-qPCR, for transcriptome analysis of arterial and venous grafts. In the search for potential molecular factors, we analyzed gene ontologies of different expression based on the muscular system. Among interesting groups, we distinguished muscle cell proliferation, muscle contraction, muscle system process, regulation of smooth muscle cell proliferation and smooth muscle cell proliferation. The highest increase in gene expression was observed in: ACTN2, RBPMS2, NR4A3, KCNA5, while the smallest decrease in expression was shown by the P2RX1, KCNH2, DES and MYOT genes. Particularly noteworthy are the ACTN2 and NR4A3 genes, which can have a significant impact on vascular patency. ACTN2 is a gene that can affect the formation of atherosclerotic plaques, while NR4A3 occurs in 4 of the 5 ontological groups discussed and can affect the inflammatory process in the blood vessel. To summarize, the presented study provided valuable insight into the molecular aspects characterizing the vessels used in CABG, and in particular identified genes that may be the target for further studies on duct patency. Running title: CABG grafts’ molecular analysis of ‘muscle system process’","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"8 1","pages":"12 - 23"},"PeriodicalIF":0.0,"publicationDate":"2020-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44877298","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}