Search

Linea Blichert-Refsgaard,1 Mette Nørgaard,2 Maria Bisgaard Bengtsen,2 Jørgen Bjerggaard Jensen1 1Department of Urology, Aarhus University Hospital, Aarhus, Denmark; 2Department of Clinical Epidemiology, Aarhus University Hospital, Aarhus, DenmarkCorrespondence: Linea Blichert-Refsgaard, Department of Urology, Aarhus University Hospital, Palle Juul-Jensens Boulevard 35, Aarhus, 8200, Denmark, Tel +45 30915431, Email lineblic@rm.dkPurpose: Globally non-muscle invasive bladder cancer (NMIBC) is a high-incidence disease. There is a large heterogeneity within NMIBC regarding recurrence- and progression risks, and large-scale studies of treatment patterns and prognoses in an everyday setting could result in NMIBC-subgroup treatment optimization, benefiting both patients and the economy. The Danish national registries provide such an opportunity if the registered procedure codes are valid. Therefore, the aim of the study was to validate the International Classification of Diseases, 10th Edition (ICD-10) codes of NMIBC treatment used in the Danish National Patient Registry (DNPR).Patients and Methods: From the DNPR, we randomly selected 200 NMIBC treatment courses identified by the dates of the course and the codes of transurethral resection of the bladder ((TURB), n = 125), photodynamic diagnosis ((PDD), n = 25), bladder instillation with Bacillus Calmette vaccine ((BCG), n = 25), or bladder instillation with chemotherapy/Mitomycin C ((MMC), n = 25). We used medical record reviews as the reference standard and estimated positive predictive values (PPVs) of all procedure codes and negative predictive values (NPVs) of PDD- and the perioperative single-shot MMC codes.Results: We identified the medical records in 150 (75%) of the 200 treatment courses (149 individual patients). The overall PPVs were TURB: 98.9% (95% confidence interval: 93.8; 100.0%), PDD: 95.8% (78.9; 99.9%), adjuvant BCG: 90.0% (68.3; 98.8%), perioperative single-shot MMC 1/5, and adjuvant MMC: 69.2% (38.6; 90.9%). The overall NPVs were PDD: 64.8% (54.4; 73.9%) and perioperative single-shot MMC: 97.7% (92.1; 99.4%).Conclusion: The ICD-10 NMIBC procedure codes recorded in the DNPR are generally valid with high PPVs. The NPV of the PPD code is acceptable. However, the code for perioperative single-shot MMC is uncertain with low PPV, but a high NPV.Keywords: validation, procedure codes, non-muscle invasive bladder cancer, Danish National Patient Registry

Maria Bisgaard Bengtsen,1 Uffe Heide-Jørgensen,1 Linea Sandfeld Blichert-Refsgaard,2 Thomas Johannesson Hjelholt,1 Michael Borre,2 Mette Nørgaard1 1Department of Clinical Epidemiology, Aarhus University Hospital, Aarhus N, Denmark; 2Department of Urology, Aarhus University Hospital, Aarhus N, DenmarkCorrespondence: Maria Bisgaard BengtsenDepartment of Clinical Epidemiology, Aarhus University Hospital, Olof Palmes Allé 43-45, Aarhus N, DenmarkTel +45 87155872Fax +45 87167215Email mariabb@clin.au.dkBackground and Aim: Benign prostatic hyperplasia comprises a significant burden to ageing men due to frequently associated lower urinary tract symptoms and the risk of developing serious complications, such as acute urinary retention. Healthcare databases are a valuable source of epidemiological research; however, continuous validation of definitions is imperative. We examined the positive predictive values of International Classification of Diseases, 10th Revision (ICD-10), diagnostic coding for benign prostatic hyperplasia and acute urinary retention in men in the Danish National Patient Registry.Methods: We investigated a random sample of 100 men diagnosed with benign prostatic hyperplasia and 100 men diagnosed with acute urinary retention between 2011 and 2017 in the Central Denmark Region. Using medical record review as reference standard, we estimated the positive predictive value with corresponding 95% confidence intervals (CI) overall and stratified by age, type of hospital (university hospital vs regional hospital), type of hospital contact (inpatient, outpatient or emergency room), calendar year group (2011– 2013, 2014– 2017), and department (department of urology, geriatrics, endocrinology or emergency room).Results: Medical records were available for all 200 sampled patients. We found an overall positive predictive value (PPV) of 95% (95% CI: 89– 98%) for benign prostatic hyperplasia and 98% (95% CI: 93– 99%) for acute urinary retention. The PPVs were consistent across age, type of hospital, type of hospital contact, calendar year group, and department.Conclusion: The PPVs of ICD-10 codes for benign prostatic hyperplasia and acute urinary retention recorded in the Danish National Patient Registry are high.Keywords: epidemiology, benign prostatic hyperplasia, acute urinary retention, validity

Lysine methylation is an abundant posttranslational modification, which has been most intensively studied in the context of histone proteins, where it represents an important epigenetic mark. Lysine methylation of histone proteins is primarily catalyzed by SET-domain methyltransferases (MTases). However, it has recently become evident that also another MTase family, the so-called seven-β-strand (7BS) MTases, often denoted METTLs (methyltransferase-like), contains several lysine (K)-specific MTases (KMTs). These enzymes catalyze the attachment of up to three methyl groups to lysine residues in specific substrate proteins, using S-adenosylmethionine (AdoMet) as methyl donor. About a decade ago, only a single human 7BS KMT was known, namely the histone-specific DOT1L, but 15 additional 7BS KMTs have now been discovered and characterized. These KMTs typically target a single nonhistone substrate that, in most cases, belongs to one of the following three protein groups: components of the cellular protein synthesis machinery, mitochondrial proteins, and molecular chaperones. This article provides an extensive overview and discussion of the human 7BS KMTs and their biochemical and biological roles., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Context: Iatrogenic hypoglycemia remains one of the leading hindrances of optimal glycemic management in insulin-treated diabetes. Recurring hypoglycemia leads to a condition of hypoglycemia-associated autonomic failure (HAAF). HAAF refers to a combination of (i) impaired hormonal counterregulatory responses and (ii) hypoglycemia unawareness to subsequent hypoglycemia, substantially increasing the risk of severe hypoglycemia. Several studies since the 1990s have experimentally induced HAAF, yielding variable results., Objective: The aim of this review was to assess the varying designs, clinical outcomes, potential assets, and drawbacks related to these studies., Method: A systemic literature search was conducted on PubMed and Embase in winter 2021 to include all human studies attempting to experimentally induce HAAF. In different combinations, the search terms used were "hypoglycemia-associated autonomic failure," "HAAF," "hypoglycemia," "recurring," "recurrent," "repeated," "consecutive," and "unawareness," yielding 1565 publications. Inclusion criteria were studies that had aimed at experimentally inducing HAAF and measuring outcomes of hormonal counterregulation and awareness of hypoglycemia., Results: The literature search yielded 27 eligible publications, of which 20 were successful in inducing HAAF while statistical significantly impairing both hormonal counterregulation and impairing awareness of hypoglycemia to subsequent hypoglycemia. Several factors were of significance as regards inducing HAAF: Foremost, the duration of antecedent hypoglycemia should be at least 90 minutes and blood glucose should be maintained below 3.4 mmol/L. Other important factors to consider are the type of participants, insulin dosage, and the risk of unintended hypoglycemia prior to the study., Conclusion: Here we have outlined the most important factors to take into consideration when designing a study aimed at inducing HAAF, including to take into consideration other disease states susceptible to hypoglycemia, thus hopefully clarifying the field and allowing qualified studies in the future., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Endocrine Society.)

Muscle cells have different phenotypes adapted to different usage, and can be grossly divided into fast/glycolytic and slow/oxidative types. While most muscles contain a mixture of such fiber types, we aimed at providing a genome-wide analysis of the epigenetic landscape by ChIP-Seq in two muscle extremes, the fast/glycolytic extensor digitorum longus (EDL) and slow/oxidative soleus muscles. Muscle is a heterogeneous tissue where up to 60% of the nuclei can be of a different origin. Since cellular homogeneity is critical in epigenome-wide association studies we developed a new method for purifying skeletal muscle nuclei from whole tissue, based on the nuclear envelope protein Pericentriolar material 1 (PCM1) being a specific marker for myonuclei. Using antibody labelling and a magnetic-assisted sorting approach, we were able to sort out myonuclei with 95% purity in muscles from mice, rats and humans. The sorting eliminated influence from the other cell types in the tissue and improved the myo-specific signal. A genome-wide comparison of the epigenetic landscape in EDL and soleus reflected the differences in the functional properties of the two muscles, and revealed distinct regulatory programs involving distal enhancers, including a glycolytic super-enhancer in the EDL. The two muscles were also regulated by different sets of transcription factors; e.g. in soleus, binding sites for MEF2C, NFATC2 and PPARA were enriched, while in EDL MYOD1 and SIX1 binding sites were found to be overrepresented. In addition, more novel transcription factors for muscle regulation such as members of the MAF family, ZFX and ZBTB14 were identified., Competing Interests: The authors have declared that no competing interests exist.

In recent years the role of altered alpha cell function and glucagon secretion in type 1 diabetes has attracted scientific attention. It is well established that glucagon responses to hypoglycemia are absent in type 1 diabetes, but more uncertain whether it is intact following other physiological and metabolic stimuli compared with nondiabetic individuals. The aim of this review is to (i) summarize current knowledge on glucagon responses during hypoglycemia in normal physiology and type 1 diabetes, and (ii) review human in vivo studies investigating glucagon responses after other stimuli in individuals with type 1 diabetes and nondiabetic individuals. Available data suggest that in type 1 diabetes the absence of glucagon secretion after hypoglycemia is irreversible. This is a scenario specific to hypoglycemia, since other stimuli, including administration of amino acids, insulin withdrawal, lipopolysaccharide exposure and exercise lead to substantial glucagon responses though attenuated compared to nondiabetic individuals in head-to-head studies. The derailed glucagon secretion is not confined to hypoglycemia as individuals with type 1 diabetes, as opposed to nondiabetic individuals display glucagon hypersecretion after meals, thereby potentially contributing to insulin resistance. The complexity of these phenomena may relate to activation of distinct regulatory pathways controlling glucagon secretion i.e., intra-islet paracrine signaling, direct and autonomic nervous signaling., (© 2021 The Authors. Physiological Reports published by Wiley Periodicals LLC on behalf of The Physiological Society and the American Physiological Society.)

Context: Women show an accelerated loss of muscle mass around menopause, possibly related to the decline in estrogen. Furthermore, the anabolic response to resistance exercise seems to be hampered in postmenopausal women., Objective: We aimed to test the hypothesis that transdermal estrogen therapy (ET) amplifies the skeletal muscle response to resistance training in early postmenopausal women., Design: A double-blinded randomized controlled study., Setting: Department of Public Health, Aarhus University, Denmark., Participants: Thirty-one healthy, untrained postmenopausal women no more than 5 years past menopause., Interventions: Supervised resistance training with placebo (PLC, n = 16) or transdermal ET ( n = 15) for 12 weeks., Main Outcome Measures: The primary outcome parameter was a cross-sectional area of quadriceps femoris measured by magnetic resonance imaging, and secondary parameters were fat-free mass (dual-energy X-ray absorptiometry), muscle strength, and functional tests., Results: The increase in muscle cross-sectional area was significantly greater in the ET group (7.9%) compared with the PLC group (3.9%) ( p < 0.05). Similarly, the increase in whole-body fat-free mass was greater in the ET group (5.5%) than in the PLC group (2.9%) ( p < 0.05). Handgrip strength increased in ET ( p < 0.05) but did not change in the PLC group. Muscle strength parameters, jumping height, and finger strength were all improved after the training period with no difference between groups., Conclusion: The use of transdermal ET enhanced the increase in muscle mass in response to 12 weeks of progressive resistance training in early postmenopausal women., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Dam, Dalgaard, Ringgaard, Johansen, Bisgaard Bengtsen, Mose, Lauritsen, Ørtenblad, Gravholt and Hansen.)

The small ubiquitin-like modifier (SUMO) post-translationally modifies lysine residues of transcription factors and co-regulators and thereby contributes to an important layer of control of the activities of these transcriptional regulators. Likewise, deSUMOylation of these factors by the sentrin-specific proteases (SENPs) also plays a role in gene regulation, but whether SENPs functionally interact with other regulatory factors that control gene expression is unclear. In the present work, we focused on SENP1, specifically, on its role in activation of gene expression investigated through analysis of the SENP1 interactome, which revealed that SENP1 physically interacts with the chromatin remodeler chromodomain helicase DNA-binding protein 3 (CHD3). Using several additional methods, including GST pulldown and co-immunoprecipitation assays, we validated and mapped this interaction, and using CRISPR-Cas9-generated CHD3- and SENP1-KO cells (in the haploid HAP1 cell line), we investigated whether these two proteins are functionally linked in regulating chromatin remodeling and gene expression. Genome-wide ATAC-Seq analysis of the CHD3- and SENP1-KO cells revealed a large degree of overlap in differential chromatin openness between these two mutant cell lines. Moreover, motif analysis and comparison with ChIP-Seq profiles in K562 cells pointed to an association of CHD3 and SENP1 with CCCTC-binding factor (CTCF) and SUMOylated chromatin-associated factors. Lastly, genome-wide RNA-Seq also indicated that these two proteins co-regulate the expression of several genes. We propose that the functional link between chromatin remodeling by CHD3 and deSUMOylation by SENP1 uncovered here provides another level of control of gene expression., (© 2018 Rodríguez-Castañeda et al.)

Context: Acromegaly has been associated with increased risk of cancer morbidity and mortality, but research findings remain conflicting and population-based data are scarce. We therefore examined whether patients with acromegaly are at higher risk of cancer., Design: A nationwide cohort study (1978 to 2010) including 529 acromegaly cases was performed. Incident cancer diagnoses and mortality were compared with national rates estimating standardized incidence ratios (SIRs). A meta-analysis of cancer SIRs from 23 studies (including the present one) was performed., Results: The cohort study identified 81 cases of cancer after exclusion of cases diagnosed within the first year [SIR 1.1; 95% confidence interval (CI), 0.9 to 1.4]. SIRs were 1.4 (95% CI, 0.7 to 2.6) for colorectal cancer, 1.1 (95% CI, 0.5 to 2.1) for breast cancer, and 1.4 (95% CI, 0.6 to 2.6) for prostate cancer. Whereas overall mortality was elevated in acromegaly (SIR 1.3; 95% CI, 1.1 to 1.6), cancer-specific mortality was not. The meta-analysis yielded an SIR of overall cancer of 1.5 (95% CI, 1.2 to 1.8). SIRs were elevated for colorectal cancer, 2.6 (95% CI, 1.7 to 4.0); thyroid cancer, 9.2 (95% CI, 4.2 to 19.9); breast cancer, 1.6 (1.1 to 2.3); gastric cancer, 2.0 (95% CI, 1.4 to 2.9); and urinary tract cancer, 1.5 (95% CI, 1.0 to 2.3). In general, cancer SIR was higher in single-center studies and in studies with <10 cancer cases., Conclusions: Cancer incidence rates were slightly elevated in patients with acromegaly in our study, and this finding was supported by the meta-analysis of 23 studies, although it also suggested the presence of selection bias in some earlier studies.

Background: Recent large-scale undertakings such as ENCODE and Roadmap Epigenomics have generated experimental data mapped to the human reference genome (as genomic tracks) representing a variety of functional elements across a large number of cell types. Despite the high potential value of these publicly available data for a broad variety of investigations, little attention has been given to the analytical methodology necessary for their widespread utilisation., Findings: We here present a first principled treatment of the analysis of collections of genomic tracks. We have developed novel computational and statistical methodology to permit comparative and confirmatory analyses across multiple and disparate data sources. We delineate a set of generic questions that are useful across a broad range of investigations and discuss the implications of choosing different statistical measures and null models. Examples include contrasting analyses across different tissues or diseases. The methodology has been implemented in a comprehensive open-source software system, the GSuite HyperBrowser. To make the functionality accessible to biologists, and to facilitate reproducible analysis, we have also developed a web-based interface providing an expertly guided and customizable way of utilizing the methodology. With this system, many novel biological questions can flexibly be posed and rapidly answered., Conclusions: Through a combination of streamlined data acquisition, interoperable representation of dataset collections, and customizable statistical analysis with guided setup and interpretation, the GSuite HyperBrowser represents a first comprehensive solution for integrative analysis of track collections across the genome and epigenome. The software is available at: https://hyperbrowser.uio.no., (© The Author 2017. Published by Oxford University Press.)

LIM-domain proteins, containing multiple cysteine-rich zinc finger-like motifs, have been shown to play diverse roles in several cellular processes. A common theme is that they mediate important protein-protein interactions that are key to their function. Androgen receptor-associated protein 55 (ARA55) belongs to this family of bridging proteins containing four C-terminal LIM domains. It has a dual role with functions both at focal adhesions and in the nucleus, apparently shuttling between the two compartments. In the present work, we have expanded our understanding of its nuclear functions by showing that it interacts with three nuclear regulators not previously linked to ARA55. We first identified ARA55 as a novel interaction partner of the nuclear kinase HIPK1 and found that ARA55, like HIPK1, also interacts with the transcription factor c-Myb. In search of a function for these associations, we observed that the coactivator p300 not only binds to c-Myb, but to ARA55 as well. When combined, c-Myb, p300, HIPK1 and ARA55 caused strong synergistic activation of a chromatinized reporter gene. In parallel, all partners, including p300, were efficiently recruited to chromatin at the c-Myb-bound promoter. Consistent with this cooperation, we found that c-Myb and ARA55 share a common set of target genes in an osteosarcoma cellular context. We propose that ARA55 and HIPK1 assist c-Myb in recruiting the coactivator and acetyltransferase p300 to chromatin., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Objective: Illicit use of growth hormone (GH) as a performance-enhancing drug among athletes is prevalent, although the evidence of such effects in healthy, young subjects is sparse. We therefore performed a meta-analysis of published studies on the effect of GH administration on body composition, substrate metabolism, and athletic performance in healthy, young subjects., Design: The English-language based databases PubMed, EMBASE, and Cochrane Central Register of Controlled Trials were searched, and eligible articles were reviewed in accordance with the PRISMA guidelines. Fifty-four potentially relevant articles were retrieved of which 11 were included in this analysis comprising 254 subjects., Results: Administration of GH significantly increased lean body mass (p<0.01) and decreased fat mass (p<0.01). In addition, GH increased the exercising levels of glycerol (p=0.01) and free fatty acids (p<0.01), but did not alter the respiratory quotient during exercise (p=0.30). GH significantly increased anaerobic exercise capacity (p<0.01) in the only study which investigated this, but did not over weeks to months improve muscle strength (p=0.36) or maximum oxygen uptake (p=0.89)., Conclusion: GH administration elicits significant changes in body composition, but does not increase either muscle strength or aerobic exercise capacity in healthy, young subjects., (Copyright © 2017. Published by Elsevier Ltd.)

The PIAS proteins (Protein Inhibitor of Activated STATs) constitute a family of multifunctional nuclear proteins operating as SUMO E3 ligases and being involved in a multitude of interactions. They participate in a range of biological processes, also beyond their well-established role in the immune system and cytokine signalling. They act both as transcriptional corepressors and coactivators depending on the context. In the present work, we investigated mechanisms by which PIAS1 causes activation or repression of c-Myb dependent target genes. Analysis of global expression data shows that c-Myb and PIAS1 knockdowns affect a subset of common targets, but with a dual outcome consistent with a role of PIAS1 as either a corepressor or coactivator. Our mechanistic studies show that PIAS1 engages in a novel interaction with the acetyltransferase and coactivator p300. Interaction and ChIP analysis suggest a bridging function where PIAS1 enhances p300 recruitment to c-Myb-bound sites through interaction with both proteins. In addition, the E3 activity of PIAS1 enhances further its coactivation. Remarkably, the SUMO status of c-Myb had a decisive role, indicating a SUMO-dependent switch in the way PIAS1 affects c-Myb, either as a coactivator or corepressor. Removal of the two major SUMO-conjugation sites in c-Myb (2KR mutant), which enhances its activity significantly, turned PIAS1 into a corepressor. Also, p300 was less efficiently recruited to chromatin by c-Myb-2KR. We propose that PIAS1 acts as a "protein inhibitor of activated c-Myb" in the absence of SUMOylation while, in its presence, PIAS behaves as a "protein activator of repressed c-Myb"., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Strict control of tissue-specific gene expression plays a pivotal role during lineage commitment. The transcription factor c-Myb has an essential role in adult haematopoiesis and functions as an oncogene when rearranged in human cancers. Here we have exploited digital genomic footprinting analysis to obtain a global picture of c-Myb occupancy in the genome of six different haematopoietic cell-types. We have biologically validated several c-Myb footprints using c-Myb knockdown data, reporter assays and DamID analysis. We show that our predicted conserved c-Myb footprints are highly dependent on the haematopoietic cell type, but that there is a group of gene targets common to all cell-types analysed. Furthermore, we find that c-Myb footprints co-localise with active histone mark H3K4me3 and are significantly enriched at exons. We analysed co-localisation of c-Myb footprints with 104 chromatin regulatory factors in K562 cells, and identified nine proteins that are enriched together with c-Myb footprints on genes positively regulated by c-Myb and one protein enriched on negatively regulated genes. Our data suggest that c-Myb is a transcription factor with multifaceted target regulation depending on cell type.