Effects of Cancer Presence and Therapy on the Platelet Proteome

Platelets are involved in tumor angiogenesis and cancer progression. Previous studies indicated that cancer could affect platelet content. In the current study, we investigated whether cancer-associated proteins can be discerned in the platelets of cancer patients, and whether antitumor treatment may affect the platelet proteome. Platelets were isolated from nine patients with different cancer types and ten healthy volunteers. From three patients, platelets were isolated before and after the start of antitumor treatment. Mass spectrometry-based proteomics of gel-fractionated platelet proteins were used to compare patients versus controls and before and after treatment initiation. A total of 4059 proteins were detected, of which 50 were significantly more abundant in patients, and 36 more in healthy volunteers. Eight of these proteins overlapped with our previous cancer platelet proteomics study. From these data, we selected potential biomarkers of cancer including six upregulated proteins (RNF213, CTSG, PGLYRP1, RPL8, S100A8, S100A9) and two downregulated proteins (GPX1, TNS1). Antitumor treatment resulted in increased levels of 432 proteins and decreased levels of 189 proteins. In conclusion, the platelet proteome may be affected in cancer patients and platelets are a potential source of cancer biomarkers. In addition, we found in a small group of patients that anticancer treatment significantly changes the platelet proteome.     

Antithrombin and Its Role in Host Defense and Inflammation

Antithrombin (AT) is a natural anticoagulant that interacts with activated proteases of the coagulation system and with heparan sulfate proteoglycans (HSPG) on the surface of cells. The protein, which is synthesized in the liver, is also essential to confer the effects of therapeutic heparin. However, AT levels drop in systemic inflammatory diseases. The reason for this decline is consumption by the coagulation system but also by immunological processes. Aside from the primarily known anticoagulant effects, AT elicits distinct anti-inflammatory signaling responses. It binds to structures of the glycocalyx (syndecan-4) and further modulates the inflammatory response of endothelial cells and leukocytes by interacting with surface receptors. Additionally, AT exerts direct antimicrobial effects: depending on AT glycosylation it can bind to and perforate bacterial cell walls. Peptide fragments derived from proteolytic degradation of AT exert antibacterial properties. Despite these promising characteristics, therapeutic supplementation in inflammatory conditions has not proven to be effective in randomized control trials. Nevertheless, new insights provided by subgroup analyses and retrospective trials suggest that a recommendation be made to identify the patient population that would benefit most from AT substitution. Recent experiment findings place the role of various AT isoforms in the spotlight. This review provides an overview of new insights into a supposedly well-known molecule.     

Nitroreductase-Mediated Release of Inhibitors of Lysine-Specific Demethylase 1 (LSD1) from Prodrugs in Transfected Acute Myeloid Leukaemia Cells

Lysine-specific demethylase 1 (LSD1) has evolved as a promising therapeutic target for cancer treatment, especially in acute myeloid leukaemia (AML). To approach the challenge of site-specific LSD1 inhibition, we developed an enzyme-prodrug system with the bacterial nitroreductase NfsB (NTR) that was expressed in the virally transfected AML cell line THP1-NTR⁺. The cellular activity of the NTR was proven with a new luminescent NTR probe. We synthesised a diverse set of nitroaromatic prodrugs that by design do not affect LSD1 and are reduced by the NTR to release an active LSD1 inhibitor. The emerging side products were differentially analysed using negative controls, thereby revealing cytotoxic effects. The 2-nitroimidazolyl prodrug of a potent LSD1 inhibitor emerged as one of the best prodrug candidates with a pronounced selectivity window between wild-type and transfected THP1 cells. Our prodrugs are selectively activated and release the LSD1 inhibitor locally, proving their suitability for future targeting approaches.     

A toy or a friend? Children's anthropomorphic beliefs about robots and how these relate to second-language word learning

This study investigates the degree to which children anthropomorphize a robot tutor and whether this anthropomorphism relates to their vocabulary learning in a second-language (L2) tutoring intervention. With this aim, an anthropomorphism questionnaire was administered to 5-year-old children (N = 104) twice: prior to and following a seven-session L2 vocabulary training with a humanoid robot. On average, children tended to anthropomorphize the robot prior to and after the lessons to a similar degree, but many children changed their attributed anthropomorphic features. Boys anthropomorphized the robot less after the lessons than girls. Moreover, there was a weak but significant positive correlation between anthropomorphism as measured before the lessons and scores on a word-knowledge post-test administered the day after the last lesson. There was also a weak but significant positive correlation between the change in anthropomorphism over time and scores on a word-knowledge post-test administered approximately 2 weeks after the last lesson. Our results underscore the need to manage children's expectations in robot-assisted education. Also, future research could explore adaptations to individual children's expectations in child-robot interactions.     

System-oriented ecotoxicological research: which way to go?

This paper presents an overview of the possibilities for further development of tools and. approaches for the ecological assessment and management of diffusively contaminated ecosystems. It is based on the results of the “Netherlands Stimulation Programme on Ecosystem-oriented Ecotoxicological Research”, the SSEO programme, which ran from 1998 to 2006, and on opinions of international experts on ecological and ecotoxicological risk-assessment methods and their legal applications. The paper also discusses the pros and cons of the set-up of the SSEO programme.Proper management of diffusively polluted areas has to be based on an integral risk-based and system approach. The approach has to be founded on the relationships between pollution, natural stresses, management measures and the presence and activities of specific species. Furthermore, the relationships between biodiversity in ecosystems and its stability and functioning have to be known. The assessment of aquatic ecosystem quality is now based on the comparisons of the composition of actual species with that of reference species. This type of system does not yet exist for the assessment of soil quality, but it is being developed. It is shown that ecological quality criteria based on a Species Sensitivity Distributions approach are sufficiently conservative to avoid or prevent major ecological impacts of diffuse pollution at concentrations below legal standards.However, a proper quality relationship of biodiversity and ecosystem functioning is lacking in the ecological assessment methods. Future research should focus on the relationship of ecosystem structure (species composition) and ecosystem functioning and on the impact of disturbing the environment and appropriate management measures.     

Immunostimulatory CpG oligonucleotides form defined three-dimensional structures: results from an NMR study

The DNA eicosamer 5'-TCCATGACGTTCCTGATGCT-3' is known to stimulate the innate immune system of vertebrae. The immunostimulatory activity is based on the activation of Toll-like receptor 9 (TLR9). While it is known that the CG dinucleotide of the eicosamer has to be unmethylated, the structural basis of the recognition of the DNA through the receptor remains unclear. Oligodeoxynucleotides containing the sequence of the eicosamer, or a portion thereof, ranging in length from hexamer to pentaeicosamer were studied by (1)H NMR spectroscopy. Based on two-dimensional NMR spectra, a number of resonances could be unambiguously assigned. For all oligonucleotides, structural transitions were detected upon heating, as monitored by the line width and chemical shift of low-field resonances. This includes the TC dinucleotide of the 5'-terminal portion, which does not have any clear base-pairing partners. The melting transitions, together with the NOESY cross-peaks, demonstrate that structure formation occurs well beyond the core hexamer 5'-GACGTT-3', a fact that may be important for understanding the molecular recognition by the Toll-like receptors of the innate immune system.     

Native protein nanolithography that can write, read and erase

The development of systematic approaches to explore protein-protein interactions and dynamic protein networks is at the forefront of biological sciences. Nanopatterned protein arrays offer significant advantages for sensing applications, including short diffusion times, parallel detection of multiple targets and the requirement for only tiny amounts of sample. Atomic force microscopy (AFM) based techniques have successfully demonstrated patterning of molecules, including stable proteins, with submicrometre resolution. Here, we introduce native protein nanolithography for the nanostructured assembly of even fragile proteins or multiprotein complexes under native conditions. Immobilized proteins are detached by a novel vibrational AFM mode (contact oscillation mode) and replaced by other proteins, which are selectively self-assembled from the bulk. This nanolithography permits rapid writing, reading and erasing of protein arrays in a versatile manner. Functional protein complexes may be assembled with uniform orientation at dimensions down to 50 nm. Such fabrication of two-dimensionally arranged nano-objects with biological activity will prove powerful for proteome-wide interaction screens and single molecule/virus/cell analyses.