Structural basis for recruitment of DAPK1 to the KLHL20 E3 ligase

BTB-Kelch proteins form the largest subfamily of Cullin-RING E3 ligases, yet their substrate complexes are mapped and structurally characterized only for KEAP1 and KLHL3. KLHL20 is a related CUL3-dependent ubiquitin ligase linked to autophagy, cancer, and Alzheimer's disease that promotes the ubiquitination and degradation of substrates including DAPK1, PML, and ULK1. We identified an “LPDLV”-containing motif in the DAPK1 death domain that determines its recruitment and degradation by KLHL20. A 1.1-Å crystal structure of a KLHL20 Kelch domain-DAPK1 peptide complex reveals DAPK1 binding as a loose helical turn that inserts deeply into the central pocket of the Kelch domain to contact all six blades of the β propeller. Here, KLHL20 forms salt-bridge and hydrophobic interactions including tryptophan and cysteine residues ideally positioned for covalent inhibitor development. The structure highlights the diverse binding modes of β-propeller domains versus linear grooves and suggests a new target for structure-based drug design. KLHL20 is a BTB-Kelch family E3 ligase linked to autophagy, cancer, and Alzheimer's disease. Chen et al. identify an “LPDLV” motif in DAPK1 that determines its recruitment and degradation by KLHL20. A 1.1-Å crystal structure of a KLHL20-DAPK1 complex reveals a hydrophobic pocket in KLHL20 suitable for inhibitor development.</p

Two paths to let the replisome go

Accurate DNA replication is essential for genome mainte- nance. Two recent reports have uncovered new molecular mechanisms controlling the termination phase of DNA replication in higher eukaryotes and established crucial roles for the CRL2LRR1 ubiquitin ligase and the p97 segregase in replisome unloading from chromatin

Efficiency measures of emergency departments: an Italian systematic literature review

Life expectancy globally increased in the last decades: the number of people aged 65 or older is consequently projected to grow, and healthcare demand will increase as well. In the recent years, the number of patients visiting the hospital emergency departments (EDs) rocked in almost all countries of the world. These departments are crucial in all healthcare systems and play a critical role in providing an efficient assistance to all patients. A systematic literature review covering PubMed, Scopus and the Cochrane Library was performed from 2009 to 2019. Of the 718 references found in the literature research, more than 25 studies were included in the current review. Different predictors were associated with the quality of EDs care, which may help to define and implement preventive strategies in the near future. There is no harmonisation in efficiency measurements reflecting the performance in the ED setting. The identification of consistent measures of efficiency is crucial to build an evidence base for future initiatives. The aim of this study is to review the literature on the problems encountered in the efficiency of EDs around the world in order to identify an organisational model or guidelines that can be implemented in EDs to fill inefficiencies and ensure access optimal treatment both in terms of resources and timing. This review will support policy makers to improve the quality of health facilities, and, consequently of the entire healthcare systems

Proteomic analysis of cell cycle progression in asynchronous cultures, including mitotic subphases, using PRIMMUS

AbstractThe temporal regulation of protein abundance and post-translational modifications is a key feature of cell division. Recently, we analysed gene expression and protein abundance changes during interphase under minimally perturbed conditions (Ly et al. 2014; Ly et al. 2015). Here we show that by using specific intracellular immunolabeling protocols, FACS separation of interphase and mitotic cells, including mitotic subphases, can be combined with proteomic analysis by mass spectrometry. Using this PRIMMUS (PRoteomic analysis of Intracellular iMMUnolabeled cell Subsets) approach, we now compare protein abundance and phosphorylation changes in interphase and mitotic fractions from asynchronously growing human cells. We identify a set of 115 phosphorylation sites increased during G2, which we term ‘early risers’. This set includes phosphorylation of S738 on TPX2, which we show is important for TPX2 function and mitotic progression. Further, we use PRIMMUS to provide a proteome-wide analysis of protein abundance remodeling between prophase, prometaphase and anaphase.</jats:p

Molecular mechanisms of cell death:recommendations of the Nomenclature Committee on Cell Death 2018

Over the past decade, the Nomenclature Committee on Cell Death (NCCD) has formulated guidelines for the definition and interpretation of cell death from morphological, biochemical, and functional perspectives. Since the field continues to expand and novel mechanisms that orchestrate multiple cell death pathways are unveiled, we propose an updated classification of cell death subroutines focusing on mechanistic and essential (as opposed to correlative and dispensable) aspects of the process. As we provide molecularly oriented definitions of terms including intrinsic apoptosis, extrinsic apoptosis, mitochondrial permeability transition (MPT)-driven necrosis, necroptosis, ferroptosis, pyroptosis, parthanatos, entotic cell death, NETotic cell death, lysosome-dependent cell death, autophagy-dependent cell death, immunogenic cell death, cellular senescence, and mitotic catastrophe, we discuss the utility of neologisms that refer to highly specialized instances of these processes. The mission of the NCCD is to provide a widely accepted nomenclature on cell death in support of the continued development of the field.</p

Health Technology Assessment on the use of the Wearable Cardioverter Defibrillator in Patients with Myocardial Infarction and with ICD Explant

The objective of the present work is to conduct a Health Technology Assessment (HTA) on the use of the Wearable Cardioverter Defibrillator (WCD) in patients at risk of Sudden Cardiac Arrest (SCA) following Myocardial Infarction (MI) or with an explanted Implantable Cardioverter Defibrillator (ICD)

Cost Minimization Analysis of Radiofrequency Compared to Laser Thermal Ablation in Patients with Hepatocellular Carcinoma

BACKGROUND: Over the last decade of years, minimally invasive techniques have been developed for the treatment of hepatocellular carcinoma and liver metastases. We sought to investigate the health costs associated with the management of patients with hepatocellular carcinoma treated with radiofrequency vs laser thermal ablation and their clinical outcomes.METHODS: We performed a retrospective analysis of the ablations performed in two referral centers in southern Italy, from 2009 to 2013. Resource use was valued by year 2017 official prices, in €. Direct healthcare costs (drugs, visits, tests and hospitalizations) of different ablation techniques were compared. Total costs were analyzed from Italian NHS perspective.RESULTS: A total of 140 patients were identified. Baseline demographics and clinical outcomes of interest did not differ between the two groups. Patients treated with laser thermal ablation resulted in an expected annually cost savings of 258.9 € per patient, in one-year follow-up healthcare costs compared with radiofrequency. The largest components of annual medical expenditures were attributable to drugs, regardless of the type of ablative technique.CONCLUSIONS: The ablation using either laser thermal ablation or radiofrequency is equally effective. Laser thermal ablation would carry disposable cost savings as compared to radiofrequency. The costs associated with management of patients with hepatocellular carcinoma, treated with laser thermal ablation were lower than those treated with radiofrequency ablation