Journal of Circulating Biomarkers最新文献_第8页

Journal of Circulating Biomarkers Pub Date : 2016-01-22 eCollection Date: 2016-01-01 DOI: 10.5772/62219

Evo K Lindersson Søndergaard, Lotte Hatting Pugholm, Rikke Bæk, Malene Møller Jørgensen, Anne Louise Schacht Revenfeld, Kim Varming

{"title":"Oxygen-Related Differences in Cellular and Vesicular Phenotypes Observed for Ovarian Cell Cancer Lines.","authors":"Evo K Lindersson Søndergaard, Lotte Hatting Pugholm, Rikke Bæk, Malene Møller Jørgensen, Anne Louise Schacht Revenfeld, Kim Varming","doi":"10.5772/62219","DOIUrl":"10.5772/62219","url":null,"abstract":"Extracellular vesicles (EVs) are one of several tools that cells use to communicate with each other. This communication is facilitated by a number of surface-associated proteins and the cargo of the vesicles. For several cancer types, the amount of EVs is observed to be up-regulated in patients compared to healthy individuals, possibly signifying the presence of an aberrant process. The hypoxia-induced release of EVs from cancer cells has been hypothesized to cause the malignant transformation of healthy recipient cells. In this study, the phenotype of cells and EVs from the ovarian cancer cell lines, COV504, SKOV3, and Pt4, were quantified and analysed under normoxic and hypoxic conditions. It was shown that both cells and EVs express common markers and that the EV phenotype varies more than the cellular phenotype. Additionally, cells subjected to 24 hours of hypoxia compared to normoxia produced more EVs. The phenotyping of EVs from cancer cell lines provides information about their molecular composition. This information may be translated to knowledge regarding the functionality of EVs and lead to a better understanding of their role in cancer.","PeriodicalId":37524,"journal":{"name":"Journal of Circulating Biomarkers","volume":"5 ","pages":"1"},"PeriodicalIF":0.0,"publicationDate":"2016-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5772/62219","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35436350","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

Detection and Characterization of Circulating Tumour Cells in Multiple Myeloma. 多发性骨髓瘤循环肿瘤细胞的检测与定性

Journal of Circulating Biomarkers Pub Date : 2016-01-01 DOI: 10.5772/64124

Liangxuan Zhang, Sharon Beasley, Natalie L Prigozhina, Renee Higgins, Shoji Ikeda, Florence Y Lee, Dena Marrinucci, Shidong Jia

{"title":"Detection and Characterization of Circulating Tumour Cells in Multiple Myeloma.","authors":"Liangxuan Zhang, Sharon Beasley, Natalie L Prigozhina, Renee Higgins, Shoji Ikeda, Florence Y Lee, Dena Marrinucci, Shidong Jia","doi":"10.5772/64124","DOIUrl":"10.5772/64124","url":null,"abstract":"Multiple myeloma (MM) remains an incurable disease despite recent therapeutic improvements. The ability to detect and characterize MM circulating tumour cells (CTCs) in peripheral blood provides an alternative to replace or augment invasive bone marrow (BM) biopsies with a simple blood draw, providing real-time, clinically relevant information leading to improved disease management and therapy selection. Here we have developed and qualified an enrichment-free, cell-based immunofluorescence MM CTC assay that utilizes an automated digital pathology algorithm to distinguish MM CTCs from white blood cells (WBCs) on the basis of CD138 and CD45 expression levels, as well as a number of morphological parameters. These MM CTCs were further characterized for expression of phospho-ribosomal protein S6 (pS6) as a readout for PI3K/AKT pathway activation. Clinical feasibility of the assay was established by testing blood samples from a small cohort of patients, where we detected populations of both CD138pos and CD138neg MM CTCs. In this study, we developed an immunofluorescent cell-based assay to detect and characterize CTCs in MM.","PeriodicalId":37524,"journal":{"name":"Journal of Circulating Biomarkers","volume":"5 ","pages":"10"},"PeriodicalIF":0.0,"publicationDate":"2016-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5548310/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35535118","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

Protein Detection Using the Multiplexed Proximity Extension Assay (PEA) from Plasma and Vaginal Fluid Applied to the Indicating FTA Elute Micro Card™. 使用多路接近扩展试验(PEA)从血浆和阴道液中检测蛋白质，用于指示FTA洗脱微卡™。

Journal of Circulating Biomarkers Pub Date : 2016-01-01 DOI: 10.5772/64000

Malin Berggrund, Daniel Ekman, Inger Gustavsson, Karin Sundfeldt, Matts Olovsson, Stefan Enroth, Ulf Gyllensten

{"title":"Protein Detection Using the Multiplexed Proximity Extension Assay (PEA) from Plasma and Vaginal Fluid Applied to the Indicating FTA Elute Micro Card™.","authors":"Malin Berggrund, Daniel Ekman, Inger Gustavsson, Karin Sundfeldt, Matts Olovsson, Stefan Enroth, Ulf Gyllensten","doi":"10.5772/64000","DOIUrl":"https://doi.org/10.5772/64000","url":null,"abstract":"The indicating FTA elute micro card™ has been developed to collect and stabilize the nucleic acid in biological samples and is widely used in human and veterinary medicine and other disciplines. This card is not recommended for protein analyses, since surface treatment may denature proteins. We studied the ability to analyse proteins in human plasma and vaginal fluid as applied to the indicating FTA elute micro card™ using the sensitive proximity extension assay (PEA). Among 92 proteins in the Proseek Multiplex Oncology Iv2 panel, 87 were above the limit of detection (LOD) in liquid plasma and 56 among 92 above LOD in plasma applied to FTA cards. Washing and protein elution protocols were compared to identify an optimal method. Liquid-based cytology samples showed a lower number of proteins above LOD than FTA cards with vaginal fluid samples applied. Our results demonstrate that samples applied to the indicating FTA elute micro card™ are amendable to protein analyses, given that a sensitive protein detection assay is used. The results imply that biological samples applied to FTA cards can be used for DNA, RNA and protein detection.","PeriodicalId":37524,"journal":{"name":"Journal of Circulating Biomarkers","volume":"5 ","pages":"9"},"PeriodicalIF":0.0,"publicationDate":"2016-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5772/64000","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35535117","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

The Role of Isolation Methods on a Nanoscale Surface Structure and its Effect on the Size of Exosomes. 分离方法对纳米级表面结构的作用及其对外泌体大小的影响

Journal of Circulating Biomarkers Pub Date : 2016-01-01 DOI: 10.5772/64148

JungReem Woo, Shivani Sharma, James Gimzewski

{"title":"The Role of Isolation Methods on a Nanoscale Surface Structure and its Effect on the Size of Exosomes.","authors":"JungReem Woo, Shivani Sharma, James Gimzewski","doi":"10.5772/64148","DOIUrl":"10.5772/64148","url":null,"abstract":"Exosomes are ∼100 nanometre diameter vesicles secreted by mammalian cells. These emerging disease biomarkers carry nucleic acids, proteins and lipids specific to the parental cells that secrete them. Exosomes are typically isolated in bulk by ultracentrifugation, filtration or immunoaffinity precipitation for downstream proteomic, genomic, or lipidomic analysis. However, the structural properties and heterogeneity of isolated exosomes at the single vesicle level are not well characterized due to their small size. In this paper, by using high-resolution atomic force microscope imaging, we show the nanoscale morphology and structural heterogeneity in exosomes derived from U87 cells. Quantitative assessment of single exosomes reveals nanoscale variations in morphology, surface roughness and counts isolated by ultracentrifugation (UC) and immunoaffinity (IA) purification. Both methods produce intact globular, 30-120 nm sized vesicles when imaged under fluid and in air. However, IA exosomes had higher surface roughness and bimodal size population compared to UC exosomes. The study highlights the differences in size and surface topography of exosomes purified from a single cell type using different isolation methods.","PeriodicalId":37524,"journal":{"name":"Journal of Circulating Biomarkers","volume":"5 ","pages":"11"},"PeriodicalIF":0.0,"publicationDate":"2016-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5548320/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35427686","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

The Most Favourable Procedure for the Isolation of Cell-Free DNA from the Plasma of Iso-Immunized RHD-Negative Pregnant Women. 从等免疫RHD阴性孕妇血浆中分离无细胞DNA的最理想程序

Journal of Circulating Biomarkers Pub Date : 2015-12-09 eCollection Date: 2015-01-01 DOI: 10.5772/62113

Riyaz Ahmad Rather, Subhas Chandra Saha, Veena Dhawan

{"title":"The Most Favourable Procedure for the Isolation of Cell-Free DNA from the Plasma of Iso-Immunized RHD-Negative Pregnant Women.","authors":"Riyaz Ahmad Rather, Subhas Chandra Saha, Veena Dhawan","doi":"10.5772/62113","DOIUrl":"10.5772/62113","url":null,"abstract":"Background: The ability to achieve quality recovery of cell-free foetal DNA is important for making non-invasive prenatal diagnoses. In this study, we performed quantitative and qualitative analyses of isolated DNA from maternal plasma, using different DNA-isolation methods.Method: DNA was isolated from 30 iso-immunized women via the QIAamp column-based method, using four different elution volumes and two conventionally based methods. Real-time polymerase chain-reaction quantification of RHD and β-globin genes was performed in order to determine foetal-specific sequences and total genome equivalents, respectively.Results: The column-based method at a 3 μl elution volume yielded the highest quality and quantity of total DNA (67.0±0.6 ng/μL). At a 3 μl elution volume, the β-globin and RHD-gene sequences were estimated to be the highest among all isolation procedures, with 2778.13±1.5 and 66.9±0.6 GEq/mL, respectively, and a 100% sensitivity for RHD-gene sequence detection. Among the two conventional manual methods, the boiling lysis method yielded a higher DNA concentration (53.8±0.8 ng/μL) and purity (1.73±0.05). In addition, the method's sensitivity for foetal-detection sequences was only 80%, whereas the salting-out method's sensitivity was just 70%.Conclusions: This study confirms the theory that the QIAamp method is a specific and sensitive approach for purifying and quantifying plasma DNA, when used in the minimum elution volume.","PeriodicalId":37524,"journal":{"name":"Journal of Circulating Biomarkers","volume":"4 ","pages":"12"},"PeriodicalIF":0.0,"publicationDate":"2015-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5548194/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35436348","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

An Immunofluorescence-Assisted Microfluidic Single Cell Quantitative Reverse Transcription Polymerase Chain Reaction Analysis of Tumour Cells Separated from Blood. 免疫荧光辅助微流控单细胞定量反转录聚合酶链反应分析血液中分离的肿瘤细胞。

Journal of Circulating Biomarkers Pub Date : 2015-11-02 eCollection Date: 2015-01-01 DOI: 10.5772/61822

Kazunori Hoshino, HaeWon Chung, Chun-Hsien Wu, Kaarthik Rajendran, Yu-Yen Huang, Peng Chen, Konstantin V Sokolov, Jonghwan Kim, John X J Zhang

{"title":"An Immunofluorescence-Assisted Microfluidic Single Cell Quantitative Reverse Transcription Polymerase Chain Reaction Analysis of Tumour Cells Separated from Blood.","authors":"Kazunori Hoshino, HaeWon Chung, Chun-Hsien Wu, Kaarthik Rajendran, Yu-Yen Huang, Peng Chen, Konstantin V Sokolov, Jonghwan Kim, John X J Zhang","doi":"10.5772/61822","DOIUrl":"https://doi.org/10.5772/61822","url":null,"abstract":"Circulating tumour cells (CTCs) are important indicators of metastatic cancer and may provide critical information for individualized treatment. As CTCs are usually very rare, the techniques to obtain information from very small numbers of cells are crucial. Here, we propose a method to perform a single cell quantitative reverse transcription polymerase chain reaction (qPCR) analysis of rare tumour cells. We utilized a microfluidic immunomagnetic assay to separate cancer cells from blood. A combination of detailed immunofluorescence and laser microdissection enabled the precise selection of individual cells. Cancer cells that were spiked into blood were successfully separated and picked up for a single cell PCR analysis. The breast cancer cell lines MCF7, SKBR3 and MDAMB231 were tested with 10 different genes. The result of the single cell analysis matched the results from a few thousand cells. Some markers (e.g., ER, HER2) that are commonly used for cancer identification showed relatively large deviations in expression levels. However, others (e.g., GRB7) showed deviations that are small enough to supplement single cell disease profiling.","PeriodicalId":37524,"journal":{"name":"Journal of Circulating Biomarkers","volume":"4 ","pages":"11"},"PeriodicalIF":0.0,"publicationDate":"2015-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5772/61822","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35436347","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: 6

Influence of Pre-Analytical Factors on Thymus- and Activation-Regulated Chemokine Quantitation in Plasma. 分析前因素对胸腺和激活调节的血浆趋化因子定量的影响。

Journal of Circulating Biomarkers Pub Date : 2015-10-27 eCollection Date: 2015-01-01 DOI: 10.5772/61749

Xuemei Zhao, Liliana Delgado, Russell Weiner, Omar F Laterza

{"title":"Influence of Pre-Analytical Factors on Thymus- and Activation-Regulated Chemokine Quantitation in Plasma.","authors":"Xuemei Zhao, Liliana Delgado, Russell Weiner, Omar F Laterza","doi":"10.5772/61749","DOIUrl":"https://doi.org/10.5772/61749","url":null,"abstract":"Thymus- and activation-regulated chemokine (TARC) in serum/plasma associates with the disease activity of atopic dermatitis (AD), and is a promising tool for assessing the response to the treatment of the disease. TARC also exists within platelets, with elevated levels detectable in AD patients. We examined the effects of pre-analytical factors on the quantitation of TARC in human EDTA plasma. TARC levels in platelet-free plasma were significantly lower than those in platelet-containing plasma. After freeze-thaw, TARC levels increased in platelet-containing plasma, but remained unchanged in platelet-free plasma, suggesting TARC was released from the platelets during the freeze-thaw process. In contrast, TARC levels were stable in serum independent of freeze-thaw. These findings underscore the importance of pre-analytical factors to TARC quantitation. Plasma TARC levels should be measured in platelet-free plasma for accurate quantitation. Pre-analytical factors influence the quantitation, interpretation, and implementation of circulating TARC as a biomarker for the development of AD therapeutics.","PeriodicalId":37524,"journal":{"name":"Journal of Circulating Biomarkers","volume":"4 ","pages":"10"},"PeriodicalIF":0.0,"publicationDate":"2015-10-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5772/61749","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35436346","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

Quantification of Cell-Free HER-2 DNA in Plasma from Breast Cancer Patients: Sensitivity for Detection of Metastatic Recurrence and Gene Amplification. 乳腺癌患者血浆中无细胞HER-2 DNA的定量:检测转移复发和基因扩增的敏感性

Journal of Circulating Biomarkers Pub Date : 2015-08-31 eCollection Date: 2015-01-01 DOI: 10.5772/61320

Patricia Diana Sørensen, Rikke Fredslund Andersen, Niels Pallisgaard, Jonna Skov Madsen, Erik Hugger Jakobsen, Ivan Brandslund

{"title":"Quantification of Cell-Free HER-2 DNA in Plasma from Breast Cancer Patients: Sensitivity for Detection of Metastatic Recurrence and Gene Amplification.","authors":"Patricia Diana Sørensen, Rikke Fredslund Andersen, Niels Pallisgaard, Jonna Skov Madsen, Erik Hugger Jakobsen, Ivan Brandslund","doi":"10.5772/61320","DOIUrl":"https://doi.org/10.5772/61320","url":null,"abstract":"The purpose of this study was to quantify the free-circulating plasma HER-2 DNA (cfHER-2 DNA) and to assess the ability of analysis to discriminate between patients with primary breast cancer and healthy controls in order to detect metastatic recurrence in comparison with serum HER-2 protein and also HER-2 gene amplification. The study population consisted of 100 patients with primary breast cancer and 50 healthy female donors. An additional 22 patients with metastases were subsequently included. cfHER-2 DNA was quantified with a quantitative PCR method together with a reference gene.Results: Using a cut-off of 2.5 for the ratio of the cfHER-2 DNA/reference gene, three (of 15) tissue HER-2-positive patients had a ratio >2.5 prior to the detection of metastatic disease. In the post-metastatic/pre-chemotherapy setting, 11 (of 23) tissue HER-2-positive patients with metastases had a ratio >2.5. There was no difference between absolute preoperative cfHER-2 DNA values for patients with primary breast cancer and those for healthy controls. There was no difference between cfHER-2 DNA values taken within nine months of development of the metastatic disease and the levels in patients without metastases, but there was a significant difference in the corresponding serum HER-2 protein levels in the tissue HER-2-positive patient group.Conclusion: Amplified HER-2 DNA can be detected in plasma when using a ratio between cfHER-2 DNA and a reference gene. cfHER-2 DNA could not be used to discriminate between patients with primary breast cancer and healthy controls, and could not predict the development of metastatic disease.","PeriodicalId":37524,"journal":{"name":"Journal of Circulating Biomarkers","volume":"4 ","pages":"9"},"PeriodicalIF":0.0,"publicationDate":"2015-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5772/61320","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35436344","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

Short Course in the Microbiome. 微生物组短期课程。

Journal of Circulating Biomarkers Pub Date : 2015-07-27 eCollection Date: 2015-01-01 DOI: 10.5772/61257

Kimberly Falana, Rob Knight, Camilia R Martin, Romina Goldszmid, K Leigh Greathouse, Joanne Gere, Howard Young, Winston Patrick Kuo

{"title":"Short Course in the Microbiome.","authors":"Kimberly Falana, Rob Knight, Camilia R Martin, Romina Goldszmid, K Leigh Greathouse, Joanne Gere, Howard Young, Winston Patrick Kuo","doi":"10.5772/61257","DOIUrl":"https://doi.org/10.5772/61257","url":null,"abstract":"Over the past decade, it has become evident that the microbiome is an important environmental factor that affects many physiological processes, such as cell proliferation and differentiation, behaviour, immune function and metabolism. More importantly, it may contribute to a wide variety of diseases, including cancer, inflammatory diseases, metabolic diseases and responses to pathogens. We expect that international, integrative and interdisciplinary translational research teams, along with the emergence of FDA-approved platforms, will set the framework for microbiome-based therapeutics and diagnostics. We recognize that the microbiome ecosystem offers new promise for personalized/precision medicine and targeted treatment for a variety of diseases. The short course was held as a four-session webinar series in April 2015, taught by pioneers and experts in the microbiome ecosystem, covering a broad range of topics from the healthy microbiome to the effects of an altered microbiome from neonates to adults and the long term effects as it is related to disease, from asthma to cancer. We have learned to appreciate how beneficial our microbes are in breaking down our food, fighting off infections and nurturing our immune system, and this information provides us with ideas as to how we can manipulate our microbiome to prevent certain diseases. However, given the variety of applications, there are scientific challenges, though there are very promising areas in reference to the clinical benefits of understanding more about our microbiome, whether in our gut or on our skin: the outlook is bright. A summary of the short course is presented as a meeting dispatch.","PeriodicalId":37524,"journal":{"name":"Journal of Circulating Biomarkers","volume":"4 ","pages":"8"},"PeriodicalIF":0.0,"publicationDate":"2015-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5772/61257","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35436343","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

Exosomes: Mechanisms of Uptake. 外泌体:摄取机制。

Journal of Circulating Biomarkers Pub Date : 2015-07-17 eCollection Date: 2015-01-01 DOI: 10.5772/61186

Kelly J McKelvey, Katie L Powell, Anthony W Ashton, Jonathan M Morris, Sharon A McCracken

引用次数: 320