Formalin-fixed paraffin-embedded (FFPE) tissue samples are important for genomic analysis of thyroid carcinomas, particularly for various molecularly targeted therapies. Therefore, this study developed and validated a technique for preparing FFPE tissue samples that preserves nucleic acid quality, which is fundamental for precise genomic analysis, more effectively than conventional methods. We analyzed surgically resected thyroid gland tumors, lymph node metastases, and separately fixed tumor samples to optimize formalin fixation and assess the influence of specimen type and preparation methods on nucleic acid quality. We assessed several quality indicators, including the DNA integrity number, cycle threshold ratio, RNA integrity number, and DV200. Separately fixed tumor samples consistently exhibited higher DNA and RNA quality than conventionally processed samples. Additionally, lymph node metastases often exhibit nucleic acid quality matching or exceeding that of thyroid gland tumors, highlighting their potential reliability for genomic analysis. These findings suggest the utility of various specimen types for the comprehensive genetic profiling of thyroid carcinomas. In conclusion, this study demonstrated that preparing separately fixed tumor samples is an effective method for preserving DNA and RNA quality for genomic analyses. Biopsy punches enable specimen collection at various facilities, including those without the ability to handle frozen specimens. This contributes to the development of a method for obtaining high-quality pathological samples that can be widely used in general medical practice. Moreover, lymph node metastases often exhibit nucleic acid quality equal to or superior to that of thyroid gland tumors, highlighting their potential as acceptable sources for genomic analyses.
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http://dx.doi.org/10.1007/s12022-024-09838-9 | DOI Listing |
Biochemistry
January 2025
Department of Microbiology, Cornell University, Ithaca, New York 14853-8101, United States.
Metal ions are essential for all life. In microbial cells, potassium (K) is the most abundant cation and plays a key role in maintaining osmotic balance. Magnesium (Mg) is the dominant divalent cation and is required for nucleic acid structure and as an enzyme cofactor.
View Article and Find Full Text PDFCommun Biol
January 2025
Center for Research on Genomics and Global Health, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, 20892, USA.
Single cell studies have transformed our understanding of cellular heterogeneity in disease but the need for fresh starting material can be an obstacle, especially in the context of international multicenter studies and archived tissue. We developed a protocol to obtain high-quality cells and nuclei from dissected human skeletal muscle archived in the preservative Allprotect® Tissue Reagent. After fluorescent imaging microscopy confirmed intact nuclei, we performed four protocol variations that compared sequencing metrics between cells and nuclei enriched by either filtering or flow cytometry sorting.
View Article and Find Full Text PDFEnferm Infecc Microbiol Clin (Engl Ed)
January 2025
Microbiology Service, Clinic University Hospital, INCLIVA Health Research Institute, Valencia, Spain; CIBER de Enfermedades Infecciosas, Instituto de Salud Carlos III, Madrid, Spain; Department of Microbiology, School of Medicine, University of Valencia, Valencia, Spain. Electronic address:
Introduction: The extent to which commercially available nucleic acid extraction platforms impact the magnitude of Cytomegalovirus (CMV) DNA loads measured in plasma specimens by 1st WHO standard-normalized real-time PCR assays is uncertain.
Methods: This retrospective study compares the performance of Abbott m2000sp, Qiagen QIAsymphony SP, and KingFisher Flex platforms using plasma samples from allogeneic hematopoietic stem cell transplant recipients and plasma spiked with the CMV AD169 strain. The Abbott RealTime CMV PCR assay was used for CMV DNA quantitation.
Int Immunopharmacol
January 2025
Key Laboratory of Livestock Infectious Diseases, Ministry of Education, Key Laboratory of Zoonosis, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, 120 Dongling Road, Shenyang 110866, China; The Research Unit for Pathogenic Mechanisms of Zoonotic Parasites, Chinese Academy of Medical Sciences, 120 Dongling Road, Shenyang 110866, China. Electronic address:
Tripartite motif-containing proteins (TRIMs), comprising the greatest subfamily of E3 ubiquitin ligases with approximately 80 members of this family, are widely distributed in mammalian cells. TRIMs actively participate in ubiquitination of target proteins, a type of post-translational modification associated with protein degradation and other functions. Tripartite motif-containing protein 29 (TRIM29), a member of the TRIM family, differs from other members of this family in that it lacks the RING finger structural domain containing cysteine and histidine residues that mediates DNA binding, protein-protein interactions, and ubiquitin ligase, at its N-terminus.
View Article and Find Full Text PDFTalanta
December 2024
The Affiliated Guangdong Second Provincial General Hospital of Jinan University, Guangzhou 510317, China. Electronic address:
Tuberculosis (TB) is the second deadliest infectious disease worldwide. Current TB diagnostics utilize sputum samples, which are difficult to obtain, and sample processing is time-consuming and difficult. This study developed an integrated diagnostic platform for the rapid visual detection of Mycobacterium tuberculosis (Mtb) in breath samples at the point-of-care (POC), especially in resource-limited settings.
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