Analytical Comparison of Methods for Extraction of Short Cell-Free DNA from Urine

Publication date: Available online 20 August 2019Source: The Journal of Molecular DiagnosticsAuthor(s): Amy Oreskovic, Norman D. Brault, Nuttada Panpradist, James J. Lai, Barry R. LutzUrine cell-free DNA (cfDNA) is a valuable noninvasive biomarker for detecting cancer mutation, diagnosing infectious disease (eg, tuberculosis), monitoring organ transplantation, and prenatal screening. Conventional silica DNA extraction does not efficiently capture urine cfDNA, which is dilute (ng/mL) and highly fragmented (30 to 100 nt). The clinical sensitivity of urine cfDNA detection increases with decreasing target length, motivating use of sample preparation methods designed for short fragments. We compared the analytical performance of two published protocols (Wizard/GuSCN and Q Sepharose), three commercial kits (Norgen, QIAamp, and MagMAX), and an in-house sequence-specific hybridization capture technique. Dependence on fragment length (25 to 150 nt), performance at low concentrations (10 copies/mL), tolerance to variable urine conditions, and susceptibility to PCR inhibition was characterized. Hybridization capture and Q Sepharose performed best overall (60% to 90% recovery), although Q Sepharose had reduced recovery (
Source: The Journal of Molecular Diagnostics - Category: Pathology Source Type: research

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We report a case of disseminated abdominal tuberculosis developing 10 years after kidney transplantation and review the underlying literature. Patient concerns and diagnoses: A 51-year-old lady presented with epigastric pain, diarrhea, weight loss and night sweats 10 years after deceased-donor kidney transplantation. An epigastric as well as multiple peritoneal masses were found suspicious of a cancer of unknown primary. Colonoscopy revealed a colon tumor with the biopsy showing no dysplasia but histiocytic and granulomatous infiltration with acid-fast bacilli. Mycobacterium tuberculosis was detected in the biopsy and s...
Source: Medicine - Category: Internal Medicine Tags: Research Article: Clinical Case Report Source Type: research
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Source: UCLA Newsroom: Health Sciences - Category: Universities & Medical Training Source Type: news
Discussion MDSCs violently emerge in pathological conditions in an attempt to limit potentially harmful immune and inflammatory responses. Mechanisms supporting their expansion and survival are deeply investigated in cancer, in the perspective to reactivate specific antitumor responses and prevent their contribution to disease evolution. These findings will likely contribute to improve the targeting of MDSCs in anticancer immunotherapies, either alone or in combination with immune checkpoint inhibitors. New evidence indicates that the expansion of myeloid cell differentiation in pathology is subject to fine-tuning, as its...
Source: Frontiers in Immunology - Category: Allergy & Immunology Source Type: research
Alessandro Poggi1*, Roberto Benelli2, Roberta Venè1, Delfina Costa1, Nicoletta Ferrari1, Francesca Tosetti1 and Maria Raffaella Zocchi3 1Molecular Oncology and Angiogenesis Unit, IRCCS Ospedale Policlinico San Martino, Genoa, Italy 2Immunology Unit, IRCCS Ospedale Policlinico San Martino, Genoa, Italy 3Division of Immunology, Transplantation and Infectious Diseases, IRCCS San Raffaele Scientific Institute, Milan, Italy It is well established that natural killer (NK) cells are involved in both innate and adaptive immunity. Indeed, they can recognize molecules induced at the cell surface by stress signals ...
Source: Frontiers in Immunology - Category: Allergy & Immunology Source Type: research
Conclusions Toxoplasma gondii infection induces a strong innate and adaptive immune response. While the innate immunity is important for controlling the early stages of the infection (Yarovinsky, 2014), the adaptive immunity is critical for restricting the parasite replication during the later stages (Gazzinelli et al., 1992). Amongst the adaptive immune subsets, CD8 T cells are the primary effector cells while CD4 T cells play an essential helper role to maintain long-term immunity (Casciotti et al., 2002). Notwithstanding, a robust CD8 T cell immunity induced during acute phase of infection, does not result in the total...
Source: Frontiers in cellular and infection microbiology - Category: Microbiology Source Type: research
Yong Zhao1, Peijuan Liu1, Zhiqian Xin1, Changhong Shi1, Yinlan Bai2, Xiuxuan Sun3, Ya Zhao1, Xiaoya Wang1,4, Li Liu1,5, Xuan Zhao1,4, Zhinan Chen3* and Hai Zhang1,6* 1Laboratory Animal Center, Air Force Medical University, Xi’an, China 2Department of Microbiology, Air Force Medical University, Xi’an, China 3Department of Cell Biology, National Translational Science Center for Molecular Medicine, Air Force Medical University, Xi’an, China 4College of Veterinary Medicine, Northwest A&F University, Yangling, China 5Key Laboratory for Space Bioscience and Biotechnology, School of Life Sciences, Nor...
Source: Frontiers in Genetics - Category: Genetics & Stem Cells Source Type: research
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Source: Frontiers in Immunology - Category: Allergy & Immunology Source Type: research
Giuseppe Ristagno1*, Francesca Fumagalli1, Barbara Bottazzi2, Alberto Mantovani2,3,4, Davide Olivari1, Deborah Novelli1 and Roberto Latini1 1Department of Cardiovascular Research, Mario Negri Institute for Pharmacological Research IRCCS, Milan, Italy 2Humanitas Clinical and Research Center-IRCCS, Milan, Italy 3Humanitas University, Milan, Italy 4The William Harvey Research Institute, Queen Mary University of London, London, United Kingdom The long pentraxin PTX3 is a member of the pentraxin family produced locally by stromal and myeloid cells in response to proinflammatory signals and microbial moieties. The p...
Source: Frontiers in Immunology - Category: Allergy & Immunology Source Type: research
Conclusions In the last decade, several efforts have been made to shed light into the molecular mechanisms driving the inflammatory response, in all its facets. miRNAs have been demonstrated to be pivotal players actively participating in the modulation of the early phase as well as the resolution of inflammation. Recent findings on their involvement in chronic inflammatory conditions, sepsis, and tumors strongly encourage the development of new miRNA-based therapeutic strategies. Author Contributions All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for public...
Source: Frontiers in Immunology - Category: Allergy & Immunology Source Type: research
In this study they also showed PTX3 localized in NETs formed after neutrophil activation (5). Proteomics analysis revealed that PTX3 forms complexes with two anti-microbial proteins [azurocidin (AZU1) and myeloperoxidase (MPO)] associated to NETs (30). More recently, PTX3 localization in NETs has been confirmed, and the colocalization with AZU1 and MPO has been defined more accurately (31). Further investigation will be needed to understand the involvement of PTX3 interaction with AZU1 and MPO in their antibacterial role during NET formation. Regulation of Complement Activation PTX3 interaction with microorganisms is not...
Source: Frontiers in Immunology - Category: Allergy & Immunology Source Type: research
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