Posttranscriptional Regulation of the Human LDL Receptor by the U2-Spliceosome

Background: The low-density lipoprotein receptor (LDLR) in the liver is the major determinant of LDL-cholesterol levels in human plasma. The discovery of genes that regulate the activity of LDLR helps to identify pathomechanisms of hypercholesterolemia and novel therapeutic targets against atherosclerotic cardiovascular disease.Methods: We performed a genome-wide RNA interference screen for genes limiting the uptake of fluorescent LDL into Huh-7 hepatocarcinoma cells. Top hit genes were validated by in vitro experiments as well as analyses of datasets on gene expression and variants in human populations.Results: The knockdown of 54 genes significantly inhibited LDL uptake. Fifteen of them encode for components or interactors of the U2-spliceosome. Knocking down any one of 11 out of 15 genes resulted in the selective retention of intron 3 of LDLR. The translated LDLR fragment lacks 88% of the full length LDLR and is detectable neither in non-transfected cells nor in human plasma. The hepatic expression of the intron 3 retention transcript is increased in non-alcoholic fatty liver disease as well as after bariatric surgery. Its expression in blood cells correlates with LDL-cholesterol and age. Single nucleotide polymorphisms and three rare variants of one spliceosome gene, RBM25, are associated with LDL-cholesterol in the population and familial hypercholesterolemia, respectively. Compared to overexpression of wild type RBM25, overexpression of the three rare RBM25 mutants in Huh-7 cells led to lower LDL uptake.Conclusions: We identified a novel mechanism of post-transcriptional regulation of LDLR activity in humans and associations of genetic variants of RBM25 with LDL-cholesterol levels.</p

Performance of the quick SOFA in very old ICU patients admitted with sepsis

BACKGROUND: The number of very elderly ICU patients (abbreviated to VOPs; ≥80 years) with sepsis increases. Sepsis was redefined in 2016 (sepsis 3.0) using the quick SOFA (qSOFA) score. Since then, multiple studies have validated qSOFA for prognostication in different patient categories, but the prognostic value in VOPs with sepsis is still unknown. METHODS: Retrospective cohort study including patients admitted to Dutch ICUs with sepsis, in the period 2012 to 2016, evaluating the outcome and the performance of qSOFA, an extended qSOFA model, SOFA, SAPS II and APACHE IV for hospital mortality. RESULTS: 5969 patients were included, of which 935 VOPs. Crude hospital mortality rates were 19%, 28% and 39% for patients aged 18-65, 65-80 and ≥80 years respectively. Discriminative performance of qSOFA for in-hospital mortality in VOPs was poor (AUC 0.596) and lower than that of SOFA, APACHE IV and SAPS II (0.704, 0.722 and 0.780 respectively). A qSOFA model extended with several other characteristics (AUC 0.643) was non-inferior to the full SOFA, but still inferior to APACHE IV and SAPS II, for all age groups. The Hosmer-Lemeshow goodness-of-fit test showed non-significant p-values for all models. Accuracy for both qSOFA and the extended qSOFA was lower compared to APACHE IV and SAPS II (Brier scores 0.227, 0.223, 0.184 and 0.183 respectively). CONCLUSION: The qSOFA showed worse discriminative performance to predict mortality than SOFA, APACHE IV and SAPS II in both VOPs and younger patients admitted with sepsis

Outcomes of Intensive Care Patients Older Than 90 Years: An 11‐Year National Observational Study

BACKGROUND/OBJECTIVES: Many intensive care unit (ICU) physicians are reluctant to admit patients aged 90 years and older, although evidence to support these decisions is scarce. Although the body of evidence on outcomes of patients aged 80 years and older is growing, it does not include patients aged 90 years and older. The aim of this study was to compare the short- and long-term mortality of ICU patients aged 90 years and older in the Netherlands with ICU patients aged 80 to 90 years, that is, octogenarians. DESIGN: Multicenter national cohort study over an 11-year period (2008-2018), using data of the National Intensive Care Evaluation (NICE) registry and the Dutch insurance claims registry. SETTING: All 82 ICUs in the Netherlands. PARTICIPANTS: All patients aged 80 years and older at the time of ICU admission. MEASUREMENTS: A total of 104,754 patients aged 80 years and older, of whom 9,495 (9%) were 90 years and older, were admitted to Dutch ICUs during the study period. RESULTS: ICU mortality of the patients aged 90 years and older was lower (13.8% vs 16.1%; P <.001) and hospital mortality was similar (26.1% vs 25.7%; P =.41) compared with octogenarians. After 3 months, mortality was higher for the patients aged 90 years and older (43.1% vs 33.7%; P <.001) and after 1-year mortality was 55.0% vs 42.7%; P <.001. CONCLUSION: In the Netherlands, mortality rates of patients aged 90 years and older admitted to the ICU are not as disappointing as often assumed. They have a lower ICU mortality and a similar hospital mortality compared with octogenarians. Nevertheless, their longer term mortality is higher compared with octogenarians. However, almost 3 of 4 patients leave the hospital alive, and almost half of the patients aged 90 years and older are still alive 1 year after their ICU admission. J Am Geriatr Soc 68:1842-1846, 2020

Outcomes of Intensive Care Patients Older Than 90 Years: An 11-Year National Observational Study

BACKGROUND/OBJECTIVES: Many intensive care unit (ICU) physicians are reluctant to admit patients aged 90 years and older, although evidence to support these decisions is scarce. Although the body of evidence on outcomes of patients aged 80 years and older is growing, it does not include patients aged 90 years and older. The aim of this study was to compare the short- and long-term mortality of ICU patients aged 90 years and older in the Netherlands with ICU patients aged 80 to 90 years, that is, octogenarians. DESIGN: Multicenter national cohort study over an 11-year period (2008-2018), using data of the National Intensive Care Evaluation (NICE) registry and the Dutch insurance claims registry. SETTING: All 82 ICUs in the Netherlands. PARTICIPANTS: All patients aged 80 years and older at the time of ICU admission. MEASUREMENTS: A total of 104,754 patients aged 80 years and older, of whom 9,495 (9%) were 90 years and older, were admitted to Dutch ICUs during the study period. RESULTS: ICU mortality of the patients aged 90 years and older was lower (13.8% vs 16.1%; P <.001) and hospital mortality was similar (26.1% vs 25.7%; P =.41) compared with octogenarians. After 3 months, mortality was higher for the patients aged 90 years and older (43.1% vs 33.7%; P <.001) and after 1-year mortality was 55.0% vs 42.7%; P <.001. CONCLUSION: In the Netherlands, mortality rates of patients aged 90 years and older admitted to the ICU are not as disappointing as often assumed. They have a lower ICU mortality and a similar hospital mortality compared with octogenarians. Nevertheless, their longer term mortality is higher compared with octogenarians. However, almost 3 of 4 patients leave the hospital alive, and almost half of the patients aged 90 years and older are still alive 1 year after their ICU admission. J Am Geriatr Soc 68:1842-1846, 2020

Multiple Reaction Monitoring Assay for Pre-eclampsia Related Calcyclin Peptides in Formalin Fixed Paraffin Embedded Placenta

Although the cause of pre-eclampsia during pregnancy has not been elucidated yet, it is evident that placental and maternal endothelial dysfunction is involved. We previously demonstrated that in early onset pre-eclampsia placental calcyclin (S100A6) expression is significantly higher compared to controls (De Groot, C.J.; et al. Clin. Proteomics 2007, 1, 325). In the current study, the results were confirmed and relatively quantified by using multiple reaction monitoring (MRM) on two peptide fragments of calcyclin. Cells were obtained from control (n = 5) and pre-eclamptic placental (n = 5) tissue collected by laser capture microdissection from formalin-fixed paraffin-embedded (FFPE) material treated with a solution to reverse formalin fixation. Two calcyclin peptides with an extra glycine inserted in the middle of the amino acid sequence were synthesized and used as an internal reference. Data presented show that MRM on laser microdissected material from FFPE tissue material is possible. The developed MRM assay to study quantitative levels of proteins in FFPE laser microdissected cells using nonisotopic-labeled chemical analogs of mass tagged internal references showed that in pre-eclamptic patients elevated levels of calcyclin is observed in placental trophoblast cells compared to normal trophoblast cells. By immunohistochemistry, we were able to confirm this observation in a qualitative manner

Healthcare-related costs in very elderly intensive care patients

Introduction: The long-term outcome of “very old intensive care unit patients” (VOPs; ≥ 80 years) is often disappointing. Little is known about the healthcare costs of these VOPs in comparison to younger ICU patients and the very elderly in the general population not admitted to the ICU. Methods: Data from a national health insurance claims database and a national quality registry for ICUs were combined. Costs of VOPs admitted to the ICU in 2013 were compared with costs of younger ICU patients (two groups, respectively 18–65 and 65–80 years old) and a matched control group of very elderly subjects who were not admitted to the ICU. We compared median costs and median costs per day alive in the year before ICU admission (2012), the year of ICU admission (2013) and the year after ICU admission (2014). Results: A total of 9272 VOPs were included and compared to three equally sized study groups. Median costs for VOPs in 2012, 2013 and 2014 (€5944, €35,653 and €12,565) are higher compared to the ICU 18–65 population (€3022, €30,223 and €5052, all p < 0.001) and the very elderly control population (€3590, €4238 and €4723, all p < 0.001). Compared to the ICU 65–80 population, costs of VOPs are higher in the year before and after ICU admission (€4323 and €6750, both p < 0.001), but not in the year of ICU admission (€34,448, p = 0.950). The median healthcare costs per day alive in the year before, the year of and the year after ICU admission are all higher for VOPs than for the other groups (p < 0.001). Conclusions: VOPs required more healthcare resources in the year before, the year of and the year after ICU admission compared to younger ICU patients and the very elderly control population, except compared to the ICU 65–80 population in the year of ICU admission. Healthcare costs per day alive, however, are substantially higher for VOPs than for all other study groups in all three studied years

Healthcare-related costs in very elderly intensive care patients

Introduction: The long-term outcome of “very old intensive care unit patients” (VOPs; ≥ 80 years) is often disappointing. Little is known about the healthcare costs of these VOPs in comparison to younger ICU patients and the very elderly in the general population not admitted to the ICU. Methods: Data from a national health insurance claims database and a national quality registry for ICUs were combined. Costs of VOPs admitted to the ICU in 2013 were compared with costs of younger ICU patients (two groups, respectively 18–65 and 65–80 years old) and a matched control group of very elderly subjects who were not admitted to the ICU. We compared median costs and median costs per day alive in the year before ICU admission (2012), the year of ICU admission (2013) and the year after ICU admission (2014). Results: A total of 9272 VOPs were included and compared to three equally sized study groups. Median costs for VOPs in 2012, 2013 and 2014 (€5944, €35,653 and €12,565) are higher compared to the ICU 18–65 population (€3022, €30,223 and €5052, all p < 0.001) and the very elderly control population (€3590, €4238 and €4723, all p < 0.001). Compared to the ICU 65–80 population, costs of VOPs are higher in the year before and after ICU admission (€4323 and €6750, both p < 0.001), but not in the year of ICU admission (€34,448, p = 0.950). The median healthcare costs per day alive in the year before, the year of and the year after ICU admission are all higher for VOPs than for the other groups (p < 0.001). Conclusions: VOPs required more healthcare resources in the year before, the year of and the year after ICU admission compared to younger ICU patients and the very elderly control population, except compared to the ICU 65–80 population in the year of ICU admission. Healthcare costs per day alive, however, are substantially higher for VOPs than for all other study groups in all three studied years

Auto-detection and segmentation of involved lymph nodes in HPV-associated oropharyngeal cancer using a convolutional deep learning neural network

Funding Information: Nicolette Taku received funding from the National Institutes of Health Research Education Program (R25EB025787). Kareem A. Wahid is supported by the Dr. John J. Kopchick Fellowship through The University of Texas MD Anderson UTHealth Graduate School of Biomedical Sciences, the American Legion Auxiliary Fellowship in Cancer Research, and an NIH/National Institute for Dental and Craniofacial Research (NIDCR) F31 fellowship (1 F31DE031502-01). Clifton David Fuller received funding from the National Institute for Dental and Craniofacial Research Award [1R01DE025248-01/R56DE025248] and Academic-Industrial Partnership Award [R01 DE028290], the National Science Foundation (NSF), Division of Mathematical Sciences, Joint NIH/NSF Initiative on Quantitative Approaches to Biomedical Big Data (QuBBD) Grant [NSF 1557679], the NIH Big Data to Knowledge (BD2K) Program of the National Cancer Institute (NCI) Early Stage Development of Technologies in Biomedical Computing, Informatics, and Big Data ScienceAward [1R01CA214825], the NCI Early Phase Clinical Trials in Imaging and Image-Guided Interventions Program [1R01CA218148], the NIH/NCI Cancer Center Support Grant (CCSG) Pilot Research Program Award from the UT MD Anderson CCSG Radiation Oncology and Cancer Imaging Program [P30CA016672], the NIH/NCI Head and Neck Specialized Programs of Research Excellence (SPORE) Developmental Research Program Award [P50 CA097007] and the National Institute of Biomedical Imaging and Bioengineering (NIBIB) Research Education Program [R25EB025787]. He has also received direct industry grant support from Elekta. Mohamed Naser received funding from the National Institutes of Health (R01DE028290-01). Publisher Copyright: © 2022 The Author(s)Purpose: Segmentation of involved lymph nodes on head and neck computed tomography (HN-CT) scans is necessary for the radiotherapy planning of early-stage human papilloma virus (HPV) associated oropharynx cancers (OPC). We aimed to train a deep learning convolutional neural network (DL-CNN) to segment involved lymph nodes on HN-CT scans. Methods: Ground-truth segmentation of involved nodes was performed on pre-surgical HN-CT scans for 90 patients who underwent levels II-IV neck dissection for node-positive HPV-OPC (training/validation [n = 70] and testing [n = 20]). A 5-fold cross validation approach was used to train 5 DL-CNN sub-models based on a residual U-net architecture. Validation and testing segmentation masks were compared to ground-truth masks using predetermined metrics. A lymph auto-detection model to discriminate between “node-positive” and “node-negative” HN-CT scans was developed by thresholding segmentation model outputs and evaluated using the area under the receiver operating characteristic curve (AUC). Results: In the DL-CNN validation phase, all sub-models yielded segmentation masks with median Dice ≥ 0.90 and median volume similarity score of ≥ 0.95. In the testing phase, the DL-CNN produced consensus segmentation masks with median Dice of 0.92 (IQR, 0.89–0.95), median volume similarity of 0.97 (IQR, 0.94–0.99), and median Hausdorff distance of 4.52 mm (IQR, 1.22–8.38). The detection model achieved an AUC of 0.98. Conclusion: The results from this single-institution study demonstrate the successful automation of lymph node segmentation for patients with HPV-OPC using a DL-CNN. Future studies, including validation with an external dataset, are necessary to clarify its role in the larger radiation oncology treatment planning workflow.Peer reviewe

Fetal liver X receptor activation acutely induces lipogenesis but does not affect plasma lipid response to a high-fat diet in adult mice

van Straten EM, van Meer H, Huijkman NC, van Dijk TH, Baller JF, Verkade HJ, Kuipers F, Plosch T. Fetal liver X receptor activation acutely induces lipogenesis but does not affect plasma lipid response to a high-fat diet in adult mice. Am J Physiol Endocrinol Metab 297: E1171-E1178, 2009. First published September 1, 2009; doi: 10.1152/ajpendo.00021.2009.-There is increasing evidence that the metabolic state of the mother during pregnancy affects long-term glucose and lipid metabolism of the offspring. The liver X receptors (LXR)alpha and -beta are key regulators of cholesterol, fatty acid, and glucose metabolism. LXRs are activated by oxysterols and expressed in fetal mouse liver from day 10 of gestation onward. In the present study, we aimed to elucidate whether in utero pharmacological activation of LXR would influence fetal fatty acid and glucose metabolism and whether this would affect lipid homeostasis at adult age. Exposure of pregnant mice to the synthetic LXR agonist T0901317 increased hepatic mRNA expression levels of Lxr target genes and hepatic and plasma triglyceride levels in fetuses and dams. T0901317 treatment increased absolute de novo synthesis and chain elongation of hepatic oleic acid in dams and fetuses. T0901317 exposure in utero influenced lipid metabolism in adulthood in a sex-specific manner; hepatic triglyceride content was increased (+45%) in male offspring and decreased in female offspring (-42%) when they were fed a regular chow diet compared with untreated sex controls. Plasma and hepatic lipid contents and hepatic gene expression patterns in adult male or female mice fed a high-fat diet were not affected by T0901317 pretreatment. We conclude that LXR treatment of pregnant mice induces immediate effects on lipid metabolism in dams and fetuses. Despite the profound changes during fetal life, long-term effects appeared to be rather mild and sex selective without modulating the lipid response to a high-fat diet