Fluorescence-based proteasome activity profiling

With the proteasome emerging as a therapeutic target for cancer treatment, accurate tools for monitoring proteasome (inhibitor) activity are in demand. In this chapter, we describe the synthesis and use of a fluorescent proteasome activity probe that allows for accurate profiling of proteasomal activity in cell lysates, intact cells, and murine and human patient-derived material, with high sensitivity using SDS-PAGE. The probe allows for direct scanning of the gel for fluorescent emission of the distinct proteasomal subunits and circumvents the use of Western blot analysis. Due to its suitable biochemical and biophysical properties, the fluorescent probe can also be used for confocal laser scanning microscopy and flow cytometry-based experiments

Mental health discourse and social media: Which mechanisms of cultural power drive discourse on Twitter

The global burden of mental health disorders has increased steadily during the past decade. Today, mental illness is the leading cause of total years lived with disability. At the same time, global mental health policies and budgets fall short of addressing the societal burden as mental health discourse languishes in the shadows due to stigma. As social media have become an increasingly popular source of i

The Influence of Metabolism on Drug Response in Cancer

Resistance to therapeutic agents, either intrinsic or acquired, is currently a major problem in the treatment of cancers and occurs in virtually every type of anti-cancer therapy. Therefore, understanding how resistance can be prevented, targeted and predicted becomes increasingly important to improve cancer therapy. In the last decade, it has become apparent that alterations in cellular metabolism are a hallmark of cancer cells and that a rewired metabolism is essential for rapid tumor growth and proliferation. Recently, metabolic alterations have been shown to play a role in the sensitivity of cancer cells to widely-used first-line chemotherapeutics. This suggests that metabolic pathways are important mediators of resistance toward anticancer agents. In this review, we highlight the metabolic alterations associated with resistance toward different anticancer agents and discuss how metabolism may be exploited to overcome drug resistance to classical chemotherapy

“Mental Health” as Defined by Twitter: Frames, Emotions, Stigma

This study analyzes the general public’s framing of ‘mental health’ and critically assesses the implications of these findings. A mismatch between how people think about mental health and what messages are used in mental health campaigns may hinder attempts to improve mental health awareness and reduce stigma. We have conducted frame analysis by using a combination of topic modeling and sentiment analysis, examining 10 years of mental health-related tweets (n = 695,414). The results reveal seven distinctive mental health frames: ‘Awareness’, ‘Feelings and Problematization’, ‘Classification’, ‘Accessibility and Funding’, ‘Stigma’, ‘Service’, and ‘Youth’ (arranged by salience). In analyzing these frames, we have learned that (1) the general awareness about mental health relates to mental illness, while health and well-being framing, although present, is prone to low quality of information, (2) mental health discourse is often used to problematize social issues and externalize personal anxieties, which tends toward trivialization and, possibly, treatment delays, (3) mental health discourse often revolves around popularized mental illness (e.g., depression, anxiety, but not neurocognitive diseases), (4) the mental health ‘Stigma’ frame is not overly pronounced; it revolves around violence, fear, and madness, (5) mental health is frequently politicized, especially concerning gun laws in the US and service acce

Типові схеми використання офшорних та оншорних зон для зменшення податкового навантаження бізнесу в Україні

The proteasome is able to create spliced Ags, in which two distant parts of a protein are excised and ligated together to form a novel peptide, for presentation by MHC class I molecules. These noncontiguous epitopes are generated via a transpeptidation reaction catalyzed by the proteasomal active sites. Transpeptidation reactions in the proteasome follow explicit rules and occur particularly efficiently when the C-terminal ligation partner contains a lysine or arginine residue at the site of ligation. Lysine contains two amino groups that theoretically may both participate in ligation reactions, implying that potentially not only peptide but also isopeptide linkages could be formed. Using nuclear magnetic resonance spectroscopy, we demonstrate in the present study that the proteasome can use the ε-amino group of an N-terminal lysine residue in transpeptidation reactions to create a novel type of posttranslationally modified epitopes. We show that the overall efficiency of ε ligation is only 10-fold lower as compared with α ligation, suggesting that the proteasome can produce sufficient isopeptide Ag to evoke a T cell response. Additionally, we show that isopeptides are more stable toward further proteasomal processing than are normal peptides, and we demonstrate that isopeptides can bind to HLA-A2.1 and HLA-A3 with high affinity. These properties likely increase the fraction of ε-ligated peptides presented on the cell surface for CD8+ T cell surveillance. Finally, we show that isopeptide Ags are immunogenic in vivo. We postulate that ε ligation is a genuine posttranslational modification, suggesting that the proteasome can create a novel type of Ag that is likely to play a role in immunity

Arginine deprivation enriches lung cancer proteomes with cysteine by inducing arginine-to-cysteine substitutants

Many types of human cancers suppress the expression of argininosuccinate synthase 1 (ASS1), a rate-limiting enzyme for arginine production. Although dependency on exogenous arginine can be harnessed by arginine-deprivation therapies, the impact of ASS1 suppression on the quality of the tumor proteome is unknown. We therefore interrogated proteomes of cancer patients for arginine codon reassignments (substitutants) and surprisingly identified a strong enrichment for cysteine (R&gt;C) in lung tumors specifically. Most R&gt;C events did not coincide with genetically encoded R&gt;C mutations but were likely products of tRNA misalignments. The expression of R&gt;C substitutants was highly associated with oncogenic kelch-like epichlorohydrin (ECH)-associated protein 1 (KEAP1)-pathway mutations and suppressed by intact-KEAP1 in KEAP1-mutated cancer cells. Finally, functional interrogation indicated a key role for R&gt;C substitutants in cell survival to cisplatin, suggesting that regulatory codon reassignments endow cancer cells with more resilience to stress. Thus, we present a mechanism for enriching lung cancer proteomes with cysteines that may affect therapeutic decisions.</p

Arginine deprivation enriches lung cancer proteomes with cysteine by inducing arginine-to-cysteine substitutants

Many types of human cancers suppress the expression of argininosuccinate synthase 1 (ASS1), a rate-limiting enzyme for arginine production. Although dependency on exogenous arginine can be harnessed by arginine-deprivation therapies, the impact of ASS1 suppression on the quality of the tumor proteome is unknown. We therefore interrogated proteomes of cancer patients for arginine codon reassignments (substitutants) and surprisingly identified a strong enrichment for cysteine (R&gt;C) in lung tumors specifically. Most R&gt;C events did not coincide with genetically encoded R&gt;C mutations but were likely products of tRNA misalignments. The expression of R&gt;C substitutants was highly associated with oncogenic kelch-like epichlorohydrin (ECH)-associated protein 1 (KEAP1)-pathway mutations and suppressed by intact-KEAP1 in KEAP1-mutated cancer cells. Finally, functional interrogation indicated a key role for R&gt;C substitutants in cell survival to cisplatin, suggesting that regulatory codon reassignments endow cancer cells with more resilience to stress. Thus, we present a mechanism for enriching lung cancer proteomes with cysteines that may affect therapeutic decisions.</p

Investigating Supply Chains Models and Enabling Technologies Towards Collaborative Networks

This research employs an extensive multiple case studies analysis to identify the most important business models affecting supply chain configurations and related enabling technologies towards the creation of collaborative networks. The results obtained from the investigation of 24 companies of manufacturing and process industry, informed by literature, identify four ‘design principles’ of business models, i.e. Personalized production, Servitization, Decentralized and modular production, and Recycle, Re-use and Sustainability. Each model is further described and discussed at the interplay between digitalization and collaborative network practices at supply chain level, showing that adopting one or a combination of the four design principles allows to actuate some of the most important features of collaboration like Vertical integration or networking of smart production systems, Horizontal integration through global value chain networks, Through-engineering across the entire value chain, Acceleration of manufacturing and Digitalization of products and services