Van Den Berghe Research Articles

6827 Pharmacological PPARα Activation In Combination With Ketogenic Nutrition Aggravated Muscle Weakness By Metabolic Failure In Septic Mice

Abstract Disclosure: C. Lauwers: None. M.P. Casaer: None. W. Vankrunkelsven: None. S. Derde: None. I. Derese: None. S. Vander Perre: None. P. Vermeersch: None. G.H. Van Den Berghe: None. L. Langouche: None. Introduction: In septic mice, infusion of ketone bodies (beta-hydroxybutyrate (BHB)) attenuated muscle weakness, a debilitating complication of critical illness. Interestingly, in critically ill children, fasting-induced ketosis also improved outcomes, but in adults ketosis upon fasting was impaired, likely due to suppression of PPARα, a key transcriptional regulator of ketogenesis. In septic mice, pharmacological induction of PPARα by pemafibrate (PF) alone, however, was ineffective in promoting ketosis. Therefore, we here hypothesized that PPARα activation by PF combined with ketogenic nutrition can more effectively induce ketosis and hereby reduce muscle weakness in sepsis. Methods: In a fluid-resuscitated and antibiotic-treated mouse model of prolonged (5 days) sepsis (male C57BL/6J), the impact of PF (1mg/kg/d) combined with 4 types of parenteral nutrition (PN) was studied. Mice were either allocated to PF + total PN (composed of glucose, long-chain triglycerides (LCTs) and amino acids) (TPN, n = 18), PF + TPN with an extra LCT emulsion (TPN + LCTs, n = 18), PF + a pure low dose LCT emulsion (Low LCT, n = 16) or PF + a pure high dose LCT emulsion (High LCT n = 18). Healthy control mice on standard chow served as a reference (HC, n=19). After 5 days of sepsis, ex vivo muscle force (aurora scientific®), plasma BHB and lipids (TG, FFA, LDL- and HDL-cholesterol; commercial assays), and liver gene expression levels were measured. Metabolomics of muscle tissue was assessed by LC-MS and plasma acylcarnitines by LC-MS-MS. Results: Liver PPARα gene expression was higher in all PF-treated mice than in HCs. Compared to HCs, specific muscle force was reduced in all septic mice, but mostly so in both PF + LCT groups (PF + Low LCT: 10.7%, PF + High LCT: 25.0%, PF+TPN + LCTs: 44.6%, PF+TPN: 58.4% of HC: 124.7 mN/mm²; p = 0.0001). PF + TPN + LCTs did not induce ketosis, whereas BHB plasma levels increased 95-fold in the PF + High LCT and 10-fold in the PF + Low LCT group (p < 0.0001 vs. other groups). Glycemia was lower in both PF + LCT groups relative to the other septic mice (PF + Low LCT: 52 mg/dl; PF + High LCT: 80 mg/dl; PF + TPN + LCTs: 108 mg/dl; PF + TPN: 102 mg/dl; p < 0.0001). Compared to the PF + TPN group, plasma lipids and long-chain acylcarnitines were increased in all other septic mice with the highest levels in the PF + High LCT group. Muscular glycolytic intermediates and ATP levels were depleted in both PF + LCT groups in comparison with all other septic mice and HCs. Conclusion: In septic mice, PF-induced PPARα activation combined with TPN, irrespective of the amount of LCTs, was unable to induce ketosis and did not attenuate muscle weakness. By contrast, PF-induced PPARα activation in combination with pure LCTs resulted in ketosis, but aggravated rather than improved muscle weakness, irrespective of the dose. In these groups, metabolic analyses suggested a bioenergetic failure of muscle fibers with an accumulation of plasma lipids. Presentation: 6/1/2024

6826 Mevalonate Treatment Worsened Hypocholesterolemia During Prolonged Sepsis Without Affecting Adrenal And Muscle Function

Abstract Disclosure: L. De Bruyn: None. S. Derde: None. F. Van Beek: None. S. Vander Perre: None. I. Derese: None. L. Pauwels: None. G. Van den Berghe: None. L. Langouche: None. Introduction: Sepsis is hallmarked by an immediate and sustained reduction in total-, HDL- and LDL-cholesterol (-C), associated with illness severity and a potential contributor to the development of adrenal failure and muscle weakness. The underlying mechanisms that might be involved in sepsis-induced hypocholesterolemia are multifactorial and partly understood. We hypothesized that treatment with the cholesterol precursor mevalonate might improve cholesterol availability, thereby improving adrenal gland and skeletal muscle function. Our hypothesis was tested in a validated and clinically relevant mouse model of prolonged sepsis. Methods: In a catheterized mouse model of cecal ligation and puncture-induced, fluid resuscitated and antibiotics treated polymicrobial abdominal sepsis (5 d), septic mice received continuous infusion with either mevalonate (78 mg/d) or placebo solution, whereas healthy mice served as controls (N=50). Plasma total-C (HDL- + LDL-C), CORT and total bile acids were measured, in addition to ex vivo muscle force and adrenocortical cholesterol ester content. Gene expression markers of inflammation (Tnf-α, Il-6) were measured in the adrenal gland, as well as hepatic mRNA expression of cholesterol synthesis enzymes (Hmgcs1, Hmgcr, Fdft1). Metabolites of the mevalonate pathway (mevalonic acid, squalene, lanosterol, desmosterol and (7-dehydro) cholesterol) were quantified in the liver with LC-MS. Results: The sepsis-induced reduction in plasma total-C was further decreased in mevalonate-treated septic mice (26±3 mg/dL) as compared with placebo (53±3 mg/dL) and healthy controls (108±3 mg/dL) (p<0.0001). Metabolites of the mevalonate pathway were all increased in the liver after mevalonate treatment (p<0.01), whereas hepatic cholesterol content was decreased as compared with placebo (p<0.05). Also, gene expression markers of cholesterol synthesis enzymes were further decreased in the liver of mevalonate-treated septic mice as compared with placebo (p<0.05). In the context of the unexpected further reduction in plasma total-C, no additional effect on plasma CORT, total bile acids, and sepsis-induced loss of muscle force and depletion in adrenocortical cholesterol ester content was observed. In addition, the sepsis-induced rise in markers of adrenal inflammation (Tnf-α, Il-6) was not further affected by mevalonate treatment. Conclusion: Mevalonate treatment further reduced the sepsis-induced hypocholesterolemia in prolonged septic mice, potentially due to increased feedback inhibition on hepatic cholesterol biosynthesis, but without further affecting adrenal and muscle function. Other therapies such as substitution with bovine serum HDL-C are currently investigated to improve altered cholesterol availability during sepsis. Presentation: 6/2/2024

Abstract 357: Reducing reagent-resistant transferrin receptor 1 dimer is a specific marker for ferroptosis

Abstract Ferroptosis is an iron-dependent cell death caused by the catastrophic accumulation of lipid peroxidation. Despite the burgeoning ferroptosis research, there remains a void for an intrinsic and comprehensive ferroptosis indicator. Here we reported a simple and cost-effective way to detect ferroptotic cell death using reducing reagent-resistant (RRR) transferrin receptor 1 (TfR1). The TfR1 RRR dimer strongly correlates with ferroptosis induced by various ferroptosis-inducing conditions and relies on lipid peroxidation, which is not observed in other types of cell death. Through iron-overloading and tumor xenograft mouse models, we validate the in vivo ferroptosis detection using this technique. Furthermore, we identify the presence of TfR1 RRR dimers in extracellular vesicles isolated from the plasma of mice with ferroptosis-related diseases. Since TfR1 is a ubiquitously expressed protein, the increase of TfR1 RRR dimer can be used as a specific and endogenous ferroptosis marker with potential clinical applications. Citation Format: Jianping He, Mackenzie Kelleher, Lucas Reynolds, Hannah Tanczos, Zhengrong Huang, Brett McLaughlin, Wesley Huang, Lauren Pope, Geraldine Veeckmans, Tom Vanden Berghe, Scott J. Dixon, Yatrik Shah, Zachary T. Schafer. Reducing reagent-resistant transferrin receptor 1 dimer is a specific marker for ferroptosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 357.

Abstract A004: Comparison of ctDNA detection in seven different types of body liquids from patients with metastatic breast cancer

Abstract Background. Liquid biopsies represent a less invasive alternative to tissue biopsy to characterize the disease in patients with metastatic cancer. Blood remains the most frequently investigated body liquid with regards to detection, quantification and characterization of the circulating tumor DNA (ctDNA). However, it might not capture the full disease profile, and other sources of body liquids may be complementary. The aim of the present study is to compare ctDNA detection in different types of body liquids. Patients and methods. Sixteen patients from the post-mortem tissue donation program UPTIDER (NCT04531696) were included in this study. The receptor status of their primary tumor was: estrogen receptor positive, Human Epidermal Growth Factor Receptor 2 non-amplified (ER+/HER2-) (n=12), ER-/HER2- (n=3) and ER+/HER2+ (n=1). Seven types of liquids were collected: blood, saliva, ascites, pleural fluid (PFL), cerebrospinal fluid (CSF), pericardial fluid and urine. Fluids were collected at study inclusion (blood, saliva, urine, and ascites whenever possible) and at autopsy (except for saliva). In total, 281 liquid samples were collected and processed according to standard protocols. All extracted cfDNA (supernatant), and the 16 matched germline DNA (buffy coat) samples underwent shallow whole genome sequencing. Log2 coverage ratios were computed with CNVkit, and good quality profiles were co-segmented per patient using the copynumber R package. Purity and ploidy were assessed by ABSOLUTE and manually reviewed. Associations between organ involvement and ctDNA yield were assessed by Wilcoxon rank-sum tests. Results. At the patient level, the proportion of liquid types in which ctDNA was detected was highly variable (median: 62%, IQR: 25-89%). ctDNA was detected in ascites of all patients where investigation was possible, in 92% of PFL, 77% of CSF, 69% of blood, 30% of pericardial fluid and in 10% of urine samples. No ctDNA was detected in the saliva samples. At autopsy, ctDNA could not be identified in blood in 4 out of the 16 patients but was detected in at least one of the other fluids for 3 of these patients. ctDNA levels tended to be higher in PFL and CSF in case of pleural and central nervous system (CNS) metastases, respectively. 3 patients had CSF ctDNA detected with no documented involvement of the CNS. In ascites, ctDNA levels were independent of peritoneal invasion. Conclusion. ctDNA was detected in 6 out of the 7 investigated body liquids and its level was partially associated with metastases in surrounding organs, except for ascites. In 3 patients, blood was not contributive but other liquids were, encouraging the evaluation of additional body fluids, when possible, in patients with metastatic breast cancer. These results open new avenues for the clinical monitoring and characterization of the disease. Citation Format: François Richard, Tatjana Geukens, Maxim De Schepper, Amena Mahdami, Karen Van Baelen, Marion Maetens, Ha-Linh Nguyen, Anirudh Pabba, Sophia Leduc, Edoardo Isnaldi, Maysam Mohammadzadeh Hajipirloo, Emily Vanden Berghe, Imane Bachir, Sigrid Hatse, Peter Vermeulen, Evy Vanderheyden, Bram Boeckx, Diether Lambrechts, Ann Smeets, Ines Nevelsteen, Kevin Punie, Patrick Neven, Hans Wildiers, Wouter Van Den Bogaert, Jonas Demeulemeester, Elia Biganzoli, Giuseppe Floris, Christine Desmedt. Comparison of ctDNA detection in seven different types of body liquids from patients with metastatic breast cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Translating Cancer Evolution and Data Science: The Next Frontier; 2023 Dec 3-6; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_2):Abstract nr A004.

New Insights into the Nature of the 5q- Deletion Syndrome Based on Quantitative Measurement of BAALC- Expressing Stem Cell Burdens

A discovery of nonrandom recurrent interstitial aberration at the long arm of chromosome 5 was made by Van den Berghe et al. in 1974. For a long time, this entity was classified as myelodysplastic syndrome (MDS). Meanwhile, its definition as well as classification criteria were repeatedly changed due to both clinical studies and advances in new techniques. In particular an insufficiency of ribosome-forming protein (RPS14) gene was found soon after similar gene RPS19 discovery in patients with severe inherited Diamond-Blackfan anemia (DBA). It cannot be excluded that basic pathogenetic mechanisms, including participation of activated gene TP53, seem to be similar in both entities.
 This article for the first time presents the quantitative data on the BAALC expression in the majority (25/31) of patients tested with 5q- deletions to be under the cut-off values. It concerns a group of 14/16 patients with isolated 5q- anomaly, and 11 other cases in whom 5q- deletion was combined with additional non-identical chromosomal aberrations. On the contrary, this molecular parameter exceeded the cut-off levels in all (n=10) MDS patients without 5q- abnormality. Hence, these data might effectively support an assumption of a ribosomopathy in cases of isolated 5q- deletion. Since about 8-10 % of these patients are transformed into MDS and/or secondary AML, a possible exclusion of isolated 5q- deletion syndrome from MDS category should be discussed carefully and this assumption is needed an additional support in larger studies.

THU155 Aberrant DNA Methylation Within Genes Of The HPA-axis Two Years After Paediatric Critical Illness, Role Of Glucocorticoid Treatment And Association With Impaired Physical And Neurocognitive Development.

Abstract Disclosure: G. Coppens: None. I. Vanhorebeek: None. F. Guïza: None. I. Derese: None. P.J. Wouters: None. J.F. Koen: None. S.C. Verbruggen: None. G. Van den Berghe: None. Critically ill children requiring intensive care suffer from impaired physical, neurocognitive and behavioural development two years later. Other severe early-life adverse events, such as abuse or neglect occurring during sensitive neurodevelopmental windows, have been associated with similar developmental problems. Earlier work has demonstrated the risk of impaired development of brain regions crucial for cognition and behaviour through increased glucocorticoid-signalling that, when prolonged or excessive, can induce aberrant DNA methylation within the hypothalamus-pituitary-adrenal (HPA) axis. We hypothesised that paediatric critical illness induces long-term DNA methylation changes within the HPA-axis, possibly aggravated by treatment with glucocorticoids in the paediatric intensive care unit (PICU), which may contribute to the long-term physical, neurocognitive and behavioural impairments. We assessed buccal mucosal DNA methylation (Infinium-Human Methylation-EPIC-BeadChip) of former PICU patients two years after PICU discharge (N=608 No-glucocorticoid-treatment; N=210 glucocorticoid-treatment) and matched healthy children (N=392) among key genes of the different layers of the HPA-axis. CpG-sites differentially methylated between groups were identified with use of multivariable linear regression. Differentially methylated DNA-regions were detected via clustering of differentially methylated CpG-sites using kernel estimates. These analyses were adjusted for technical variation and baseline risk factors and corrected for multiple testing (FDR<0.05). Additionally, we investigated to what extent glucocorticoid treatment during PICU stay had played a direct role in bringing about any DNA methylation changes. Finally, we assessed whether an association with the long-term physical, neurocognitive and behavioural impairment was present, with use of multivariable linear models. Former PICU patients showed aberrant DNA methylation at 10 CpG-sites (located within NR3C1, FKBP5, TSC22D3, TNF and, HSD11B1) and one DNA-region (located within TNF). Additionally, we identified that the differential methylation of 2 CpG-sites within FKBP5 was at least partly attributable to the glucocorticoid treatment that was given during PICU stay. The aberrant DNA methylation in former PICU patients associated with impaired development. This was most pronounced for the hypomethylation within the promotor/5’UTR region of FKBP5, which associated with worse growth, intelligence, visual-motor integration, alertness and memory. Two years after critical illness in children, aberrant DNA methylation within different genes of the HPA axis was detected, some of which may partly be explained by the glucocorticoid treatment given during PICU admission. These changes associated with long-term physical and neurocognitive impairments of former PICU patients. Presentation: Thursday, June 15, 2023

Children’s Digital and Non-Digital Play Practices with Cozmo, the Toy Robot

Children’s Digital and Non-Digital Play Practices with Cozmo, the Toy Robot

Abstract P5-05-06: ctDNA detection in seven different types of body liquids in patients with metastatic breast cancer

Abstract Background. Liquid biopsies represent a less invasive alternative to tissue biopsy to characterize and possibly monitor the disease in patients with metastatic breast cancer. So far, blood remains the most frequently investigated body liquid in this context and the investigations mainly focus on the detection, quantification and characterization of the circulating tumor DNA (ctDNA). However, since blood might not capture the full disease profile, other sources of body liquids may have the potential to complement the information obtained from blood. The aims of the present study are therefore to assess whether: (i) ctDNA can be detected in different types of body liquids, and, (ii) the levels of ctDNA in a given liquid are associated with metastases in specific organs. Patients and methods. Twelve patients from the post-mortem tissue donation program UPTIDER (NCT04531696) were included in this study. The receptor status of their primary tumor was: estrogen receptor negative, HER2 non-amplified (ER+/HER2-) (n=9), ER-/HER2- (n=2) and ER+/HER2+ (n=1). Median time between inclusion and death of the patient was 1.6 months (Interquartile range: [0.4-3.4]). Seven types of liquids were collected: blood, saliva, ascites, pleural fluid (PFL), cerebrospinal fluid (CSF), pericardial fluid and urine. Fluids were collected at study inclusion (blood, as well as saliva, urine, and ascites whenever possible) and at autopsy (except for saliva). In total, 108 liquid samples were collected and immediately centrifuged according to standard protocols. Cell free DNA (cfDNA) was extracted from the supernatant. All extracted cfDNA as well as germline DNA extracted from the 12 matched buffy coat samples underwent shallow whole genome sequencing. Log2 ratios were computed with CNVkit, and co-segmented per patient using the copynumber R package. Purity and ploidy were assessed by ABSOLUTE. Associations between organ involvement and ctDNA yield were assessed by Wilcoxon rank-sum tests. Samples at study inclusion and at autopsy were considered together unless otherwise specified. Results. At the sample level, ctDNA could be identified in 54% of the samples. At the patient level, the proportion of liquid types in which ctDNA was detected was highly variable (median: 58%, IQR: 34-77%, Table 1). CtDNA was detected in ascites of all patients when investigated, in 78% of PFL, 73% of CSF, 67% of blood and 37% of pericardial fluid. Only for one patient with invasive lobular carcinoma, ctDNA was detected in saliva and urine, the latter most likely explained by invasion of the bladder. Of note, in 4/12 patients ctDNA could not be identified in blood but was detected in at least one of the other fluids for 3 of these patients. At autopsy, ctDNA levels tended to be higher in PFL, ascites, and CSF in case of pleural, peritoneal, and central nervous system (CNS) metastases respectively, reaching statistical significance only for PFL. In CSF, two patients have CSF ctDNA detected with no documented involvement of the CNS. No brain autopsy was however performed for these patients. Conclusion. We have shown that ctDNA can be detected in all 7 different body liquids that were investigated in this study. The ctDNA levels in a given liquid can be associated with the presence of metastases in specific organs. Since ctDNA was not detected in 4 of our patients in blood but detectable for 3 of them in other liquids, the evaluation of additional sources of body fluids should be further investigated in patients with metastatic breast cancer. These results therefore open new avenues for the clinical monitoring and characterization of the disease. Table 1. Summary of ctDNA detection per liquid type at the patient level based on the 108 evaluated samples. Histo.= Histological, ILC= Invasive lobular carcinoma, NA= not available, nr= number, NST= non-special type Citation Format: François Richard, Tatjana Geukens, Maxim De Schepper, Amena Mahdami, Karen Van Baelen, Marion Maetens, Ha-Linh Nguyen, Anirudh Pabba, Sophia Leduc, Edoardo Isnaldi, Maysam Hajipirloo, Emily Vanden Berghe, Imane Bachir, Sigrid Hatse, Peter Vermeulen, Evy Vanderheyden, Bram Boeckx, Diether Lambrechts, Ann Smeets, Ines Nevelsteen, Kevin Punie, Patrick Neven, Hans Wildiers, Wouter Van Den Bogaert, Elia Biganzoli, Giuseppe Floris, Christine Desmedt. ctDNA detection in seven different types of body liquids in patients with metastatic breast cancer [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P5-05-06.

Abstract P6-14-14: Advancing research on metastatic breast cancer: the UPTIDER post-mortem tissue donation program

Abstract Background. Research in metastatic breast cancer is hampered by limited sample availability. Post-mortem tissue donation programs can help to overcome this problem but are logistically challenging and have thus far mainly focused on histopathological and genomic research. We here present the UPTIDER program (NCT04531696), aimed at the multilevel characterization of advanced breast cancer and generation of tumour models. Patients and Methods. Patients with stage IV breast cancer receiving their last line(s) of treatment are eligible for participation. Blood, urine and saliva samples are collected upon inclusion. Upon death, a post-mortem MRI (when possible) followed by a rapid autopsy is performed. Liquid biopsies from all body fluids and tissue samples from all macroscopically identified metastatic sites are collected. Samples are processed as mirrored biopsies in different conditions, such as fresh frozen for omics analyses, formalin fixed paraffin-embedded for histopathology, and slowly frozen in freezing medium or fresh for generation of xenograft and organoid models. Results. Since approval by the local Ethical Committee in November 2020, 22 patients have been enrolled and 15 autopsies have been performed. Mean interval between death and start of autopsy was 3h (range 2-6h), mean duration of the autopsies was 6h (4-9h). A post-mortem MRI was performed in 6 patients. Peripheral blood, central blood and bone marrow were collected from all patients; urine, ascites, cerebrospinal, pericardial and pleural fluid all in more than 2/3 of patients. On average, 232 (range 90-406) tissue samples of which 164 (45-303) pathological from 42 (15 – 79) metastases were collected for each patient. Most often sampled metastatic sites were lymph nodes, liver, bones, pleura and peritoneum. Samples from the primary tumour could be retrieved from all patients, either during the autopsy (n=6) or from historical archives. In total, 133 tumour samples were sent to collaborating partners for patient-derived xenograft creation. Already some have been successfully established and stored, including models derived from a patient with invasive lobular carcinoma (ILC) and one with metaplastic squamous cell carcinoma. When correlating microscopic and macroscopic findings, patients could largely be divided into three main categories. Eleven patients presented with overt and extensive disease burden, often characterized by diffuse visceral, pleural, peritoneal, bone and lymph node involvement. Two patients, both with ILC, presented with underestimated yet extensive disease burden. While gross examination and cross sectioning of organs did not reveal clear involvement, microscopical invasion of stomach and liver, amongst others, was found. Lastly, limited disease burden was seen in two patients, both with leptomeningeal involvement. In those patients, massive tumoral infiltration in the subarachnoid space and along the blood-brain barrier was seen microscopically, with no grey matter invasion. Conclusion. We successfully launched a new and comprehensive post-mortem tissue donation program for patients with metastatic breast cancer, enrolling ~ 1 patient per month. Post-mortem tumour samples already resulted in successful establishment of some patient-derived xenografts. From a clinical point of view, vast underestimation of the disease extent on imaging during life as well as macroscopically during the autopsy was observed in some patients with metastatic ILC. For patients with leptomeningeal metastasis, we showed that the highly aggressive nature of their disease might be explained by extensive meningeal infiltration disrupting the blood-brain barrier. Further insights into disease progression and heterogeneity will be generated by the ongoing multi-omics analyses. Citation Format: Tatjana Geukens, Maxim De Schepper, Karen Van Baelen, François Richard, Marion Maetens, Amena Mahdami, Ha-Linh Nguyen, Edoardo Isnaldi, Anirudh Pabba, Sophia Leduc, Imane Bachir, Maysam Hajipirloo, Emily Vanden Berghe, Sigrid Hatse, Eleonora Leucci, Maria Francesca Baietti, Georgios Sflomos, Cathrin Brisken, Patrick Derksen, Colinda Scheele, Vincent Vandecaveye, Ann Smeets, Ines Nevelsteen, Kevin Punie, Patrick Neven, Elia Biganzoli, Hans Wildiers, Wouter Van Den Bogaert, Giuseppe Floris, Christine Desmedt. Advancing research on metastatic breast cancer: the UPTIDER post-mortem tissue donation program [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P6-14-14.

1199 PREVENTION AND TREATMENT OF CRITICAL ILLNESS ACQUIRED WEAKNESS IN THE ELDERLY

Abstract Introduction Critical Illness Acquired weakness (ICU-Acquired Weakness (ICU-AW)) is an umbrella term used to describe Critical Illness Myopathy (CIM) and Critical Illness Polyneuropathy (CIP). The condition exerts high prevalence in the elderly admitted in the ICU and is associated with deteriorating patient outcomes, namely mortality and morbidity. The prevalence of the syndrome is highly variable in the current literature hindering our ability to objectively quantify the scale of the problem. Moreover, several preventative methods and treatment for ICU-AW as a result of sarcopenia have been proposed in literature with some of them providing favourable outcomes. Objectives 1. Evaluate the prevalence of ICU-AW in the elderly through a systematic review; and 2. Explore the treatment options currently available. Methods We conducted a systematic review using the PubMed, Embase and Cochrane databases to explore the current studies available on the diagnosis of ICU-AW syndromes. Cochrane’s Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement was our template. Results Overall, twenty-one studies (1544 patients) were included. The minimum reported prevalence is 20%, whereas the maximum is 76%. The overall median prevalence was 52% (Q1: 32% and Q3: 61%) with an interquartile range (IQR) of 29%. The highest IQR was found in studies using clinical examination (IQR=37%) whereas the lowest in studies using electrophysiological assessment (IQR= 21%). Moreover, several preventative measures for ICU-AW were identified and analyzed namely: nutritional alterations (high protein dies), glucose control, early mobilization, neuromuscular electrical stimulation and the ABCDEF bundle. Conclusion The variability in the diagnostic modalities used to measure the syndrome as well as the inconsistency in the diagnostic parameters within each modality prevent us from objectively quantifying the prevalence of ICU-AW. With regards to treatment early mobilization protocols offer promising evidence. Reference Vanhorebeek, Latronico, Van den Berghe G. ICU-acquired weakness. Intensive Care Med. 2020;46(4):637-53.

Blood glucose targets in the critically ill: is one size fits all still appropriate?

In 2001, the paradigm for blood glucose control in hospitalised patients was challenged by a single-centre, randomised controlled trial (RCT). 1 van den Berghe G Wouters P Weekers F et al. Intensive insulin therapy in critically ill patients. N Engl J Med. 2001; 345: 1359-1367 Crossref PubMed Scopus (8134) Google Scholar Conducted in mechanically ventilated surgical intensive care unit (ICU) patients (970 [63%] of 1548 following cardiovascular surgery and 1344 [87%] without a previous diagnosis of diabetes), this trial used intravenous insulin to target euglycaemia (blood glucose 80–110 mg/dL [4·4–6·1 mmol/L]) and dramatic reductions in mortality and morbidity were seen compared with patients in the control arm, in which intravenous insulin was administered only if blood glucose exceeded 215 mg/dL (11·9 mmol/L), with a target range of 180–200 mg/dL (10·0–11·1 mmol/L).

Role of Macrophages and Mast Cells as Key Players in the Maintenance of Gastrointestinal Smooth Muscle Homeostasis and Disease.

The gut contains the largest macrophage pool in the body, with populations of macrophages residing in the mucosa and muscularis propria of the gastrointestinal (GI) tract. Muscularis macrophages (MMs), which are located within the muscularis propria, interact with cells essential for GI function, such as interstitial cells of Cajal, enteric neurons, smooth muscle cells, enteric glia, and fibroblast-like cells, suggesting that these immune cells contribute to several aspects of GI function. This review focuses on the latest insights on the factors contributing to MM heterogeneity and the functional interaction of MMs with other cell types essential for GI function. This review integrates the latest findings on macrophages in other organs with increasing knowledge of MMs to better understand their role in a healthy and diseased gut. We describe the factors that contribute to (muscularis macrophage) MM heterogeneity, and the nature of MM interactions with cells regulating GI function. Finally, we also describe the increasing evidence suggesting a critical role of another immune cell type, the mast cell, in normal and diseased GI physiology.

Reviews Spring 2021

De Raedt, N. A Review of Fabrizio Nevola, Street Life in Renaissance Italy. New Haven and London: Yale University Press, 2020.Sands, F. A Review of Matthew M. Reeve, Gothic Architecture and Sexuality in the Circle of Horace Walpole. University Park, PA: The Pennsylvania State University Press, 2020.Campbell, E. A Review of Carla D’Arista, The Pucci of Florence: Patronage and Politics in Renaissance Italy. Turnhout: Harvey Miller Publishers, 2020.Vanden Berghe, V. A Review of Louise Campbell, Studio Lives: Architect, Art and Artist in 20th-Century Britain. London: Lund Humphries, 2019.

Ethnic tourism: revisiting conceptual terminology and its significance in the context of interethnic relations in conditions of globalization of the society

The article deals with the analysis of foreign and national researchers’ approaches to definition of the concept of ethnic tourism. The majority of existing studies devoted to ethnic tourism in America and Europe concentrate on describing it as trips to exotic and often isolated ethnic groups (Smith V., McIntosh R., Goeldner C., Van den Berghe P., Harron S., Weiler B.). Some scientists regard that ethnic tourism includes trips to forefathers’ homelands with the purpose of learning the own ethnic belonging – nostalgic tourism (King B.). Along with the term of ethnic tourism, the concepts of aboriginal tourism (Getz D., Jamieson W.) and native tourism (Butler R., Hinch T.) are widely used, though the same phenomenon is meant. Russian researchers use the concepts of ethnic tourism, ethnographic tourism and nostalgic tourism. No unanimous opinion as to definition of ethnic tourism is found with Ukrainian scientists (Kyryliuk L., Parkhomenko T., Kyfiak V., Bochan I., Kulakovska I., Orlova M, Chubrey O., etc). We understand the concept of ethnic tourism to be a type of tourist activity that satisfies the consumers’ interests in cognition of material and spiritual culture of certain ethnos (ethnic group, ethnographic group) that inhabits (or previously inhabited) a certain territory. While on excursions, tourists get acquainted with architecture, clothes, folklore, rituals, folk traditions, handicrafts, everyday life, ethnic cuisine, historic events, known persons, and monuments of certain ethnos or its part (sub-ethnos or ethnic group). The author discloses functions of the present-day ethnic tourism (cultural-cognitive, social, communicative, worldview, integrative) in the context of interethnic relations in conditions of globalization processes, those functions that are essentially important for the development of individuality, broadening of knowledge of the surrounding world, people, trends and regularities in the development of society with regard to the formation of value and worldview orientations.

Nwd1 Regulates Neuronal Differentiation and Migration through Purinosome Formation in the Developing Cerebral Cortex.

SummaryEngagement of neural stem/progenitor cells (NSPCs) into proper neuronal differentiation requires the spatiotemporally regulated generation of metabolites. Purines are essential building blocks for many signaling molecules. Enzymes that catalyze de novo purine synthesis are assembled as a huge multienzyme complex called “purinosome.” However, there is no evidence of the formation or physiological function of the purinosome in the brain. Here, we showed that a signal transduction ATPases with numerous domains (STAND) protein, NACHT and WD repeat domain-containing 1 (Nwd1), interacted with Paics, a purine-synthesizing enzyme, to regulate purinosome assembly in NSPCs. Altered Nwd1 expression affected purinosome formation and induced the mitotic exit and premature differentiation of NSPCs, repressing neuronal migration and periventricular heterotopia. Overexpression/knockdown of Paics or Fgams, other purinosome enzymes, in the developing brain resulted in a phenocopy of Nwd1 defects. These findings indicate that strict regulation of purinosome assembly/disassembly is crucial for maintaining NSPCs and corticogenesis.

Expert teacher perceptions of two-way feedback interaction

The importance and influence of feedback is well-established in the literature (Hattie, 2009). The purpose of feedback is to improve learning (Voerman, Meijer, Korthagen, & Simons, 2012) by reducing discrepancies (Hattie, 2007), closing gaps (Sadler, 2010), and improving one's knowledge, and skill acquisition (Moreno, 2004). However, there is disjuncture concerning the effectiveness of unilateral or one-way feedback. Unilateral feedback has been critiqued for its failure to productively engage, guide learning, and monitor performance (Price, Handley, & Millar, 2011; Sadler, 1989). Despite calls to focus feedback on student learning outcomes (Hattie, 2009; Voerman et al., 2012), a third of feedback interventions have reported a decrease in student performance (Kluger & DeNisi, 1996). Research informs that feedback that focuses on self instead of task inhibits learning (Kluger & DeNisi, 1996; Shute, 2008). On the other hand, feedback that focuses on self-regulation, task, and cognitive processing enhances learning (Hattie & Timperley, 2007; Shute, 2008). However, teachers are observed delivering one-way feedback, rather than facilitating learning (Blair & Ginty, 2013; Van den Berghe, Ros, & Beijaard, 2013), and appear ‘to close down opportunities for exploring student learning rather than opening them up’ (Torrance & Pryor, 1988, p. 621). That is, self-focussed unilateral feedbackthwarts the potential to promote learning (Burke, 2009)...

Novel mRNA-Based Therapy Reduces Toxic Galactose Metabolites and Overcomes Galactose Sensitivity in a Mouse Model of Classic Galactosemia.

Classic galactosemia (CG) is a potentially lethal inborn error of galactose metabolism that results from deleterious mutations in the human galactose-1 phosphate uridylyltransferase (GALT) gene. Previously, we constructed a GalT-/- (GalT-deficient) mouse model that exhibits galactose sensitivity in the newborn mutant pups, reduced fertility in adult females, impaired motor functions, and growth restriction in both sexes. In this study, we tested whether restoration of hepatic GALT activity alone could decrease galactose-1 phosphate (gal-1P) and plasma galactose in the mouse model. The administration of different doses of mouse GalT (mGalT) mRNA resulted in a dose-dependent increase in mGalT protein expression and enzyme activity in the liver of GalT-deficient mice. Single intravenous (i.v.) dose of human GALT (hGALT) mRNA decreased gal-1P in mutant mouse liver and red blood cells (RBCs) within 24h with low levels maintained for over a week. Repeated i.v. injections increased hepatic GalT expression, nearly normalized gal-1P levels in liver, and decreased gal-1P levels in RBCs and peripheral tissues throughout all doses. Moreover, repeated dosing reduced plasma galactose by 60% or more throughout all four doses. Additionally, a single intraperitoneal dose of hGALT mRNA overcame the galactose sensitivity and promoted the growth in a GalT-/- newborn pup.

ABRO1 promotes NLRP3 inflammasome activation through regulation of NLRP3 deubiquitination.

Deubiquitination of NLRP3 has been suggested to contribute to inflammasome activation, but the roles and molecular mechanisms are still unclear. We here demonstrate that ABRO1, a subunit of the BRISC deubiquitinase complex, is necessary for optimal NLRP3-ASC complex formation, ASC oligomerization, caspase-1 activation, and IL-1β and IL-18 production upon treatment with NLRP3 ligands after the priming step, indicating that efficient NLRP3 activation requires ABRO1. Moreover, we report that ABRO1 deficiency results in a remarkable attenuation in the syndrome severity of NLRP3-associated inflammatory diseases, including MSU- and Alum-induced peritonitis and LPS-induced sepsis in mice. Mechanistic studies reveal that LPS priming induces ABRO1 binding to NLRP3 in an S194 phosphorylation-dependent manner, subsequently recruiting the BRISC to remove K63-linked ubiquitin chains of NLRP3 upon stimulation with activators. Furthermore, deficiency of BRCC3, the catalytically active component of BRISC, displays similar phenotypes to ABRO1 knockout mice. Our findings reveal an ABRO1-mediated regulatory signaling system that controls activation of the NLRP3 inflammasome and provide novel potential targets for treating NLRP3-associated inflammatory diseases.

New fifth‐order two‐derivative Runge‐Kutta methods with constant and frequency‐dependent coefficients

Two‐derivative Runge‐Kutta methods are Runge‐Kutta methods for problems of the form y′ = f(y) that include the second derivative y′′ = g(y) = f ′(y)f(y) and were developed in the work of Chan and Tsai. In this work, we consider explicit methods and construct a family of fifth‐order methods with three stages of the general case that use several evaluations of f and g per step. For problems with oscillatory solution and in the case that a good estimate of the dominant frequency is known, methods with frequency‐dependent coefficients are used; there are several procedures for constructing such methods. We give the general framework for the construction of methods with variable coefficients following the approach of Simos. We modify the above family to derive methods with frequency‐dependent coefficients following this approach as well as the approach given by Vanden Berghe. We provide numerical results to demonstrate the efficiency of the new methods using three test problems.

Therapeutic Benefit and Gene Network Regulation by Combined Gene Transfer of Apelin, FGF2, and SERCA2a into Ischemic Heart.

Despite considerable advances in cardiovascular disease treatment, heart failure remains a public health challenge. In this context, gene therapy appears as an attractive approach, but clinical trials using single therapeutic molecules result in moderate benefit. With the objective of improving ischemic heart failure therapy, we designed a combined treatment, aimed to simultaneously stimulate angiogenesis, prevent cardiac remodeling, and restore contractile function. We have previously validated IRES-based vectors as powerful tools to co-express genes of interest. Mono- and multicistronic lentivectors expressing fibroblast growth factor 2 (angiogenesis), apelin (cardioprotection), and/or SERCA2a (contractile function) were produced and administrated by intramyocardial injection into a mouse model of myocardial infarction. Data reveal that combined treatment simultaneously improves vessel number, heart function parameters, and fibrosis prevention, due to FGF2, SERCA2a, and apelin, respectively. Furthermore, addition of SERCA2a in the combination decreases cardiomyocyte hypertrophy. Large-scale transcriptome analysis reveals that the triple treatment is the most efficient in restoring angiogenic balance as well as expression of genes involved in cardiac function and remodeling. Our study validates the concept of combined treatment of ischemic heart disease with apelin, FGF2, and SERCA2a and shows that such therapeutic benefit is mediated by a more effective recovery of gene network regulation.