ABSTRACT
XRN1 is a highly conserved exoribonuclease which degrades uncapped RNAs in a 5'-3' direction. Degradation of RNAs by XRN1 is important in many cellular and developmental processes and is relevant to human disease. Studies in D. melanogaster demonstrate that XRN1 can target specific RNAs, which have important consequences for developmental pathways. Osteosarcoma is a malignancy of the bone and accounts for 2% of all pediatric cancers worldwide. Five-year survival of patients has remained static since the 1970s and therefore furthering our molecular understanding of this disease is crucial. Previous work has shown a down-regulation of XRN1 in osteosarcoma cells; however, the transcripts regulated by XRN1 which might promote osteosarcoma remain elusive. Here, we confirm reduced levels of XRN1 in osteosarcoma cell lines and patient samples and identify XRN1-sensitive transcripts in human osteosarcoma cells. Using RNA-seq in XRN1-knockdown SAOS-2 cells, we show that 1178 genes are differentially regulated. Using a novel bioinformatic approach, we demonstrate that 134 transcripts show characteristics of direct post-transcriptional regulation by XRN1. Long noncoding RNAs (lncRNAs) are enriched in this group, suggesting that XRN1 normally plays an important role in controlling lncRNA expression in these cells. Among potential lncRNAs targeted by XRN1 is HOTAIR, which is known to be up-regulated in osteosarcoma and contributes to disease progression. We have also identified G-rich and GU motifs in post-transcriptionally regulated transcripts which appear to sensitize them to XRN1 degradation. Our results therefore provide significant insights into the specificity of XRN1 in human cells which are relevant to disease.
Subject(s)
Bone Neoplasms/genetics , Exoribonucleases/genetics , Microtubule-Associated Proteins/genetics , Osteosarcoma/genetics , RNA, Long Noncoding/genetics , RNA, Messenger/genetics , RNA, Neoplasm/genetics , Bone Neoplasms/metabolism , Bone Neoplasms/pathology , Cell Line, Tumor , Child , Computational Biology , Exoribonucleases/deficiency , Gene Expression Regulation, Neoplastic , Gene Knockdown Techniques , Genome-Wide Association Study , Humans , Microtubule-Associated Proteins/deficiency , Molecular Sequence Annotation , Nucleotide Motifs , Osteosarcoma/metabolism , Osteosarcoma/pathology , RNA Processing, Post-Transcriptional , RNA Stability , RNA, Long Noncoding/metabolism , RNA, Messenger/metabolism , RNA, Neoplasm/metabolismABSTRACT
Rotary blood pumps (RBPs) used for mechanical circulatory support of heart failure patients cannot passively change pump flow sufficiently in response to frequent variations in preload induced by active postural changes. A physiological control system that mimics the response of the healthy heart is needed to adjust pump flow according to patient demand. Thus, baseline data are required on how the healthy heart and circulatory system (i.e., heart rate (HR) and cardiac output (CO)) respond. This study investigated the response times of the healthy heart during active postural changes (supine-standing-supine) in 50 healthy subjects (27 male/23 female). Early response times (te) and settling times (ts) were calculated for HR and CO from data continuously collected with impedance cardiography. The initial circulatory response of HR and CO resulted in te of 9.0-11.7 s when standing up and te of 4.7-5.7 s when lying back down. Heart rate and CO settled in ts of 50.0-53.6 s and 46.3-58.2 s when standing up and lying down, respectively. In conclusion, when compared to active stand up, HR and CO responded significant faster initially when subjects were lying down (p < 0.05); there were no significant differences in response times between male and female subjects. These data will be used during evaluation of physiological control systems for RBPs, which may improve patient outcomes for end-stage heart failure patients.
Subject(s)
Cardiovascular Physiological Phenomena , Heart/physiology , Posture/physiology , Adult , Female , Humans , Kinetics , MaleABSTRACT
CONTEXT: It is recommended that hemoglobin (Hb) A1c (Hb A1c) not be used to assess average glycemia in patients who have altered red blood cell life span. OBJECTIVE: To investigate the frequency of reporting an Hb A1c value for Hb variant samples that do not include Hb A. DESIGN: Hb A1c samples (n = 500) were procured and screened for Hb variants that may affect Hb A1c interpretation (Hb SS, Hb SC, and Hb S-ß-thalassemia). Five of each of these samples were tested by ion-exchange high-performance liquid chromatography, immunoturbidimetric assay, second-generation immunoturbidimetric assay, and affinity chromatography. RESULTS: Eleven (2.2%) homozygous Hb SS, 6 (1.2%) Hb SC, and 5 (1.0%) Hb S-ß-thalassemia samples were identified out of the 500 samples tested. Three of 4 instruments investigated in this study are known to not be plagued by analytic interference from these Hb variants but disturbingly reported Hb A1c values in the absence of Hb A. CONCLUSIONS: The improved analytic specificity of Hb A1c platforms has by and large eliminated interferences from the most common heterozygous Hb variants. A consequence, however, is the potential for unintended reporting of Hb A1c results in the presence of homozygous and compound heterozygous Hb variants that lack Hb A and the inability to distinguish those samples not recommended to be used for patient care. The ability to identify samples harboring Hb variants that preclude the utility of Hb A1c may be beneficial in high prevalence populations.
