Matters最新文献 - Book学术

The Cell Culture Environment Regulates the Transcription Factor MafB in BV-2 Microglia. 细胞培养环境对BV-2小胶质细胞转录因子MafB的调控

Matters Pub Date : 2021-01-01 Epub Date: 2021-01-29

Patrick Miller-Rhodes, Harris A Gelbard

{"title":"The Cell Culture Environment Regulates the Transcription Factor MafB in BV-2 Microglia.","authors":"Patrick Miller-Rhodes, Harris A Gelbard","doi":"","DOIUrl":"","url":null,"abstract":"Microglia experience dramatic molecular and functional changes when transferred from the central nervous system (CNS) to a cell culture environment. Investigators largely attribute these findings to the loss of CNS-specific microenvironmental cues that dictate the gene-regulatory networks specified by master regulator transcription factors such as V-maf musculoaponeurotic fibrosarcoma oncogene homolog B (MafB). MafB regulates macrophage differentiation and activation by activating or repressing target genes critical to these processes. Here, we show that basal MafB levels in the BV-2 microglial cell line depend on the availability of lipids in the cell culture environment. Depletion of lipids, either by serum deprivation or the use of lipid-depleted serum, reduced MafB protein levels in BV-2 cells. Using live imaging, we also observed the engulfment of apoptotic BV-2 cell debris by neighboring BV-2 cells, highlighting an additional potential source of lipids in the cell culture environment. This observation was supported by experiments showing reduced MafB protein levels in BV-2 cells cultured with various phagocytosis inhibitors (cytochalasin D, annexin V) and reduced BV-2 cell phagocytic activity with serum deprivation. In aggregate, our data suggest that serum exposure regulates the transcription factor MafB in BV-2 cells through direct and indirect mechanisms.","PeriodicalId":18333,"journal":{"name":"Matters","volume":"2021 ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8101704/pdf/nihms-1677830.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38966008","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

MCV Truncated Large T antigen interacts with BRD4 in tumors. MCV截断大T抗原在肿瘤中与BRD4相互作用。

Matters Pub Date : 2019-01-11 DOI: 10.19185/matters.201811000004

Reety Arora, Arushi Vats, Vrushali Chimankar

{"title":"MCV Truncated Large T antigen interacts with BRD4 in tumors.","authors":"Reety Arora, Arushi Vats, Vrushali Chimankar","doi":"10.19185/matters.201811000004","DOIUrl":"https://doi.org/10.19185/matters.201811000004","url":null,"abstract":"Among Polyomaviridae family of viruses, Merkel Cell Polyomavirus (MCV) is the only human polyomavirus with convincing data supporting its classification as a direct causative agent of a human skin malignancy, Merkel Cell Carcinoma. Oncogenic transformation by MCV requires the integration of the viral genome into the human genome, truncation of the large T antigen (LT) to render the viral genome replication deficient and expression of small T antigen oncoprotein. The chromatin binding protein BRD4, was recently shown to transcriptionally regulate the expression of virus oncoproteins, thereby enhancing the tumorigenesis of virus-associated cancers, such as HPV associated cervical cancer. Previous work by Wang et al. revealed that BRD4 interacts with MCV full length LT during viral replication. In this study, we demonstrated that MCV truncated tumor LT antigen also interacts with BRD4 protein. We showed that the MCV tumor LT antigen and BRD4 protein complex co-localizes within the nucleus. Furthermore, we tested whether BRD4 protein transcriptionally regulates MCV Non Coding Control Region (NCCR), where we found that though full length LT and sT together, along with the BRD4 protein showed enhanced transcriptional activity whereas tumor truncated LT did not. These findings on the interactions of the MCV tumor truncated LT antigen with the BRD4 protein add to existing knowledge about interactions with LT and its role in tumorigenesis, and assist in efforts to more precisely define new therapy targets for this disease.","PeriodicalId":18333,"journal":{"name":"Matters","volume":"2019 ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7212090/pdf/EMS86188.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37925212","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Cytotoxicity of aqueous cigarette smoke extract is affected by properties of pipettes used to prepare the extract. 卷烟烟雾水提取物的细胞毒性受用于制备提取物的移液管性质的影响。

Matters Pub Date : 2019-01-01 Epub Date: 2019-10-14

Eric D Bernstein, Zhaohao Liao, Kenneth W Witwer

引用次数: 0

Lentiviral gene therapy vector with UCOE stably restores function in iPSC-derived neutrophils of a CDG patient. 慢病毒基因治疗载体UCOE稳定恢复CDG患者ipsc衍生中性粒细胞的功能。

Matters Pub Date : 2018-01-01 Epub Date: 2018-05-23 DOI: 10.19185/matters.201805000005

Walther Haenseler, Elena Kuzmenko, Adjoa Smalls-Mantey, Cathy Browne, Reinhard Seger, William S James, Sally A Cowley, Janine Reichenbach, Ulrich Siler

{"title":"Lentiviral gene therapy vector with UCOE stably restores function in iPSC-derived neutrophils of a CDG patient.","authors":"Walther Haenseler, Elena Kuzmenko, Adjoa Smalls-Mantey, Cathy Browne, Reinhard Seger, William S James, Sally A Cowley, Janine Reichenbach, Ulrich Siler","doi":"10.19185/matters.201805000005","DOIUrl":"https://doi.org/10.19185/matters.201805000005","url":null,"abstract":"A recent gamma-retroviral clinical Chronic Granulomatous Disease (CGD) gene therapy (GT) trial achieved proof-of-concept but was accompanied by activation of oncogenes and transgene silencing. The ubiquitous chromatin opening element (UCOE) comprises the sequences of two divergently oriented house-keeping gene promoters and is known to have anti-silencing properties. In a screen we identified two novel UCOE constructs that prevent adjacent promoter methylation in P19 cells. Experiments were continued with the shorter UCOE constructs in induced pluripotent stem cells (iPSC) derived from a p47phox-deficient CGD patient. The iPSC line was transduced with the lentiviral GT vectors expressing P47 under the constitutively active SFFV promoter with UCOE element (UCOE_SF) and without UCOE element (SF) adjacent to the SFFV promoter. The iPSC were expanded before propagation towards neutrophils. 20 days after transduction the UCOE_SF vector was protected from methylation in iPSC as previously shown in P19 cells, whereas the SF vector was heavily methylated in iPSC. The UCOE_SF vector maintained stable transgene expression in iPSC, macrophages and neutrophils, whereas the SF vector was strongly silenced. The UCOE_SF vector stably restored ROS production in neutrophils, whereas for the SF vector the count of ROS producing cells was marginal. To conclude, we have shown that the prevention of transgene silencing by UCOE is functionally and mechanistically preserved upon terminal neutrophil differentiation.","PeriodicalId":18333,"journal":{"name":"Matters","volume":"2018 ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6472893/pdf/nihms-1015902.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37170903","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 4

Dynasore inhibition on productive infection of HIV-1 in commonly used cell lines is independent of transferrin endocytosis. 在常用细胞系中，对HIV-1多产性感染的王朝抑制与转铁蛋白内吞作用无关。

Matters Pub Date : 2018-01-01 Epub Date: 2018-05-30 DOI: 10.19185/matters.201805000001

Hanna Song, Michael DeSantis, Chunjuan Tian, Wei Cheng

{"title":"Dynasore inhibition on productive infection of HIV-1 in commonly used cell lines is independent of transferrin endocytosis.","authors":"Hanna Song, Michael DeSantis, Chunjuan Tian, Wei Cheng","doi":"10.19185/matters.201805000001","DOIUrl":"https://doi.org/10.19185/matters.201805000001","url":null,"abstract":"The route of HIV-1 entry for productive infection in CD4+ host cells is a fundamental question for the molecular understanding of HIV-1 infection and transmission. Although direct fusion has long been thought to be the mode of entry, recent studies have suggested that productive entry of HIV-1 may actually occur through dynamin-dependent endocytosis. In several of these studies, dynasore, a noncompetitive inhibitor of the GTPase activity of dynamin, has been used to support this conclusion. Here we show that dynasore does produce inhibitory effects on the productive infection of HIV-1 in several commonly used cell lines. This effect is present regardless of the methods used to facilitate the infection of HIV-1. However, transferrin uptake remains fully functional in these cell lines upon dynasore treatment. Therefore, the inhibition on HIV-1 infection by dynasore in these cell lines is due to an effect that is independent of transferrin endocytosis. The use of dynasore in probing the role of endocytosis in HIV-1 infection should be corroborated by other methods.","PeriodicalId":18333,"journal":{"name":"Matters","volume":"2018 ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6492620/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37377621","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Structural analysis of clinically relevant pathogenic G6PD variants reveals the importance of tetramerization for G6PD activity. 临床相关致病G6PD变异的结构分析揭示了G6PD活性的四聚化的重要性。

Matters Pub Date : 2017-09-14 DOI: 10.19185/matters.201705000008

Anna D Cunningham, Daria Mochly-Rosen

{"title":"Structural analysis of clinically relevant pathogenic G6PD variants reveals the importance of tetramerization for G6PD activity.","authors":"Anna D Cunningham, Daria Mochly-Rosen","doi":"10.19185/matters.201705000008","DOIUrl":"https://doi.org/10.19185/matters.201705000008","url":null,"abstract":"Over 220 different amino acid variants have been identified in human glucose-6-phosphate dehydrogenase (G6PD), covering over 30% of the protein sequence. Many of these variants are pathogenic, causing varying degrees of G6PD deficiency with symptoms ranging from severe chronic anemia (class I) to milder triggered hemolytic episodes (classes II and III). The phenotypic effects of most G6PD variants have been reported, providing an opportunity to correlate phenotypic and structural information. In particular, we sought to investigate the tetramer interface of G6PD in relation to pathogenic variation, as there are conflicting reports indicating the importance of tetramerization for G6PD activity. Using a three-dimensional spatial scan statistic, hotspots of structural enrichment were identified for each class of pathogenic G6PD variants. Class I variants, the most phenotypically severe, were enriched at the dimer interface, consistent with previous evidence that dimerization is essential for G6PD activity. Class II variants were enriched near the tetramer interface, suggesting that tetramerization is also important for G6PD activity. This analysis explains why these two classes, both yielding 10% or less G6PD activity as compared to normal, lead to different clinical outcomes.","PeriodicalId":18333,"journal":{"name":"Matters","volume":"2017 ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2017-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6105287/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36431789","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 12

Internalization of scavenger receptor ligands by cortical neurons. 皮层神经元清道夫受体配体的内化。

Matters Pub Date : 2017-05-22 DOI: 10.19185/matters.201703000006

Chen Liang, Vrushali Chavan, Konark Mukherjee

引用次数: 0

Functional analysis of Na⁺/H⁺ exchanger 9 variants identified in patients with autism and epilepsy. 自闭症和癫痫患者Na+/H+交换物9变异的功能分析。

Matters Pub Date : 2017-04-01 Epub Date: 2017-05-22 DOI: 10.19185/matters.201704000009

Hari Prasad, James Osei-Owusu, Rajini Rao

{"title":"Functional analysis of Na+/H+ exchanger 9 variants identified in patients with autism and epilepsy.","authors":"Hari Prasad, James Osei-Owusu, Rajini Rao","doi":"10.19185/matters.201704000009","DOIUrl":"https://doi.org/10.19185/matters.201704000009","url":null,"abstract":"Na+/H+ exchanger isoform 9, NHE9, finely tunes the pH within the endosomal lumen to regulate cargo trafficking and turnover. In patients with autism, genetic approaches have revealed deletions, truncations and missense mutations in the gene encoding NHE9 (SLC9A9). To help establish causality, functional evaluation is needed to distinguish pathogenic mutations from harmless polymorphisms. Here, we evaluated three previously uncharacterized NHE9 variants, P117T, D496N, and Q609K reported in patients with autism and epilepsy. We show that NHE9-DsRed localizes to recycling endosomes in HEK293 cells where it significantly alkalinizes luminal pH, and elevates accumulation of transferrin. All three NHE9 variants were expressed and localized to endosomal compartments, similar to wild-type NHE9. In contrast to previously characterized NHE9 variants, we observed no loss-of-function with respect to endosomal pH homeostasis and transferrin endocytosis. These findings suggest that the three NHE9 substitutions analyzed in our study are either benign polymorphisms or may have a cell-type specific or regulatory function not detected in our cell culture model. Our findings highlight the importance of combining the use of cellular studies of function with sequencing technologies that capture genomic variation in patients.","PeriodicalId":18333,"journal":{"name":"Matters","volume":"2017 ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2017-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/c1/b3/nihms887306.PMC5555647.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35329185","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 8

Improving access to interventions among mothers screened positive for post-partum depression (PPD) at National Programme on Immunization (NPI) clinics in south-western and south-eastern Nigeria - A service development report. 一份服务发展报告:改善尼日利亚西南部和东南部国家免疫规划诊所产后抑郁症筛查呈阳性的母亲获得干预措施的机会。

Matters Pub Date : 2017-01-01 Epub Date: 2017-10-12 DOI: 10.19185/matters.201707000005

Muideen O Bakare, Mashudat A Bello-Mojeed, Kerim M Munir, Olaniyi O Duduyemi, Andrew O Orovwigho, Odutola I Odetunde, Olufemi G Taiwo, Jushua A Olofinlade, Olakunle N Omotoso, Olayinka H Famurewa, Oladipupo O Omolabi, Adebayo O Jejeloye

{"title":"Improving access to interventions among mothers screened positive for post-partum depression (PPD) at National Programme on Immunization (NPI) clinics in south-western and south-eastern Nigeria - A service development report.","authors":"Muideen O Bakare, Mashudat A Bello-Mojeed, Kerim M Munir, Olaniyi O Duduyemi, Andrew O Orovwigho, Odutola I Odetunde, Olufemi G Taiwo, Jushua A Olofinlade, Olakunle N Omotoso, Olayinka H Famurewa, Oladipupo O Omolabi, Adebayo O Jejeloye","doi":"10.19185/matters.201707000005","DOIUrl":"https://doi.org/10.19185/matters.201707000005","url":null,"abstract":"We investigate the possibility of improving access to interventions among mothers screened positive for post-partum depression (PPD) at National Programme on Immunization (NPI) clinics randomly selected from Lagos and Enugu States in south-western and south-eastern Nigeria respectively. The principle of human centred design was employed by engaging the mothers screened positive for PPD to be part of the decision making regarding their further assessment and intervention services. The study brought intervention services to primary healthcare centre at the NPI clinics. Improvement in willingness to seek interventions was observed among the mothers screened positive for PPD in this study when compared to our observation in a previous report, where mothers diagnosed with PPD were referred and requested to visit a mental health facility closer to their NPI clinics for further assessment and interventions (95.2% versus 33.7%). Interventional services for the mothers diagnosed with PPD also impact positively on the growth parameters of their infants on follow-up. Principle of human centred design improved access to intervention services among the mothers and infants studied. NPI clinics at primary healthcare level would provide appropriate forum for early screening of mothers for PPD and interventions in low-resource setting like Nigeria. There would be improvement in maternal and child health coverage if the Nigerian Government can adapt human centred design principles employed in this study nationwide.","PeriodicalId":18333,"journal":{"name":"Matters","volume":"2017 ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5665652/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35577887","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Mutant Cas9-transcriptional activator activates HIV-1 in U1 cells in the presence and absence of LTR-specific guide RNAs. 突变的cas9转录激活因子在存在或不存在ltr特异性引导rna的情况下激活U1细胞中的HIV-1。

Matters Pub Date : 2017-01-01 Epub Date: 2017-01-12 DOI: 10.19185/matters.201611000027

Veronica Kim, Brian M Mears, Bonita H Powell, Kenneth W Witwer

引用次数: 10