Over the past decade, laser capture microdissection (LCM) has grown as a tool for gene expression profiling of small numbers of cells from tumor samples and of specific cell populations in complex tissues. LCM can be used to study toxicant effects on selected cell populations within the testis at different stages of spermatogenesis. There are several LCM-related hurdles to overcome, including issues inherent to the method itself, as well as biases that result from amplifying the LCM-isolated RNA. Many technical issues associated with the LCM method are addressed here, including increasing RNA yield and obtaining more accurate quantification of RNA yields. We optimized the LCM method optimized to generate RNA quantities sufficient for quantitative reverse transcription polymerase chain reaction (qRT-PCR) array analysis without amplification and were able to validate the method through direct comparison of results from unamplified and amplified RNA from individual samples. The addition of an amplification step for gene expression studies using LCM RNA resulted in a bias, especially for low abundance transcripts. Although the amplification bias was consistent across samples, researchers should use caution when comparing results generated from amplified and unamplified LCM RNA. Here, we have validated the use of LCM-derived RNA with the qRT-PCR array, improving our ability to investigate cell-type and stage-specific responses to toxicant exposures.
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http://dx.doi.org/10.1177/0192623314526319 | DOI Listing |
Acta Parasitol
January 2025
Genkök Genome and Stem Cell Center, Erciyes University, Talas, Kayseri, 38039, Türkiye.
Purpose: Encephalitozoon intestinalis is an obligate intracellular microsporidian fungus that causes severe gastrointestinal infections, particularly in immunocompromised individuals. Propolis (PROP), a resinous substance derived from bees, has antimicrobial, anti-inflammatory and antioxidant properties, while royal jelly (RJ) has immunomodulatory, antioxidant and antimicrobial activities. The aim of this study was to investigate the therapeutic potential of PROP and RJ against E.
View Article and Find Full Text PDFJ Nutr
January 2025
Department of Human Physiology of the Chair of Preclinical Sciences, Medical University in Lublin, Lublin, Poland.
Background: Systemic inflammation plays a crucial role in the development and progression of chronic heart failure (CHF) across all phenotypes. The continuous release of pro-inflammatory cytokines causes muscle atrophy and adipocyte breakdown, ultimately resulting in cachexia. Long non-coding RNAs (lncRNAs) are emerging as potential biomarkers associated with cachexia, as they indirectly regulate muscle and fat tissue metabolism.
View Article and Find Full Text PDFFront Genet
January 2025
First Clinical School, Liaoning University of Traditional Chinese Medicine, Shenyang, China.
Introduction: The deficiency of estrogen correlates with a range of diseases, notably Postmenopausal osteoporosis (PMO) and Parkinson's disease (PD). There is a possibility that PMO and PD may share underlying molecular mechanisms that are pivotal in their development and progression. The objective of this study was to identify critical genes and potential mechanisms associated with PMO by examining co-expressed genes linked to PD.
View Article and Find Full Text PDFPLoS One
January 2025
Center for Inflammation, Immunity, & Infection, Institute for Biomedical Sciences, Georgia State University, Atlanta, Georgia, United States of America.
Microbiota-induced production of IL-22 by type 3 innate lymphoid cells (ILC3) plays an important role in maintaining intestinal health. Such IL-22 production is driven, in part, by IL-23 produced by gut myeloid cells that have sensed select microbial-derived mediators. The extent to which ILC3 can directly respond to microbial metabolites via IL-22 production is less clear, in part due to the difficulty of isolating and maintaining sufficient numbers of viable ILC3 ex vivo.
View Article and Find Full Text PDFNanomaterials (Basel)
December 2024
Division of Biochemical Toxicity, FDA/National Center for Toxicological Research, Jefferson, AR 72079, USA.
The safety of titanium dioxide (TiO), widely used in foods and personal care products, has been of ongoing concern. Significant toxicity of TiO has been reported, suggesting a risk to human health. To evaluate its potential epigenotoxicity, the effect of exposure to a TiO product to which humans could be exposed on DNA methylation, a primary epigenetic mechanism, was investigated using two human cell lines (Caco-2 (colorectal) and HepG2 (liver)) relevant to human exposure.
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