A new mutation-independent approach to cancer therapy : inhibiting oncogenic RAS and MYC, by targeting mitochondrial biogenesis.

Here, we used MCF7 cells as a model system to interrogate how MYC/RAS co-operativity contributes to metabolic flux and stemness in breast cancer cells. We compared the behavior of isogenic MCF7 cell lines transduced with c-Myc or H-Ras (G12V), either individually or in combination. Cancer stem cell (CSC) activity was measured using the mammosphere assay. c-Myc augmented both mammosphere formation and mitochondrial respiration, without any effects on glycolytic flux. In contrast, H-Ras (G12V) synergistically augmented both mammosphere formation and glycolysis, but only in combination with c-Myc, directly demonstrating MYC/RAS co-operativity. As c-Myc is known to exert its effects, in part, by stimulating mitochondrial biogenesis, we next examined the effects of another stimulus known to affect mitochondrial biogenesis, i.e. ROS production. To pharmacologically induce oxidative stress, we used Rotenone (a mitochondrial inhibitor) to target mitochondrial complex I. Treatment with Rotenone showed bi-phasic effects; low-dose Rotenone (1 to 2.5 nM) elevated mammosphere formation, while higher doses (10 to 100 nM) were inhibitory. Importantly, the stimulatory effects of Rotenone on CSC propagation were blocked using a mitochondrial-specific anti-oxidant, namely Mito-tempo. Thus, "mild" mitochondrial oxidative stress, originating at Complex I, was sufficient to pheno-copy the effects of c-Myc, effectively promoting CSC propagation. To validate the idea that mitochondrial biogenesis is required to stimulate CSC propagation, we employed Doxycycline, a well-established inhibitor of mitochondrial protein translation. Treatment with Doxycycline was indeed sufficient to block the stimulatory effects of H-Ras (G12V), c-Myc, and Rotenone on CSC propagation. As such, Doxycycline provides a strong rationale for designing new therapeutics to target mitochondrial biogenesis, suggesting a new "mutation-independent" approach to cancer therapy. In support of this notion, most currently successful anti-cancer agents therapeutically target "cell phenotypes", such as increased cell proliferation, rather than specific genetic mutations. Remarkably, we demonstrated that Doxycycline inhibits the effects of diverse oncogenic stimuli, of both i) genetic (MYC/RAS) and ii) environmental (Rotenone) origins. Finally, we discuss the advantages of our "Proteomics-to-Genomics (PTG)" approach for in silico validation of new biomarkers and novel drug targets. In this context, we developed a new Myc-based Mito-Signature consisting of 3 mitochondrial genes (HSPD1; COX5B; TIMM44) for effectively predicting tumor recurrence (HR=4.69; p=2.4e-08) and distant metastasis (HR=4.94; p=2.8e-07), in ER(+) in breast cancer patients. This gene signature could serve as a new companion diagnostic for the early prediction of treatment failure in patients receiving hormonal therapy

Primary ChAdOx1 vaccination does not reactivate pre-existing, cross-reactive immunity

Currently available COVID-19 vaccines include inactivated virus, live attenuated virus, mRNA-based, viral vectored and adjuvanted protein-subunit-based vaccines. All of them contain the spike glycoprotein as the main immunogen and result in reduced disease severity upon SARS-CoV-2 infection. While we and others have shown that mRNA-based vaccination reactivates pre-existing, cross-reactive immunity, the effect of vector vaccines in this regard is unknown. Here, we studied cellular and humoral responses in heterologous adenovirus-vector-based ChAdOx1 nCOV-19 (AZ; Vaxzeria, AstraZeneca) and mRNA-based BNT162b2 (BNT; Comirnaty, BioNTech/Pfizer) vaccination and compared it to a homologous BNT vaccination regimen. AZ primary vaccination did not lead to measurable reactivation of cross-reactive cellular and humoral immunity compared to BNT primary vaccination. Moreover, humoral immunity induced by primary vaccination with AZ displayed differences in linear spike peptide epitope coverage and a lack of anti-S2 IgG antibodies. Contrary to primary AZ vaccination, secondary vaccination with BNT reactivated pre-existing, cross-reactive immunity, comparable to homologous primary and secondary mRNA vaccination. While induced anti-S1 IgG antibody titers were higher after heterologous vaccination, induced CD4(+) T cell responses were highest in homologous vaccinated. However, the overall TCR repertoire breadth was comparable between heterologous AZ-BNT-vaccinated and homologous BNT-BNT-vaccinated individuals, matching TCR repertoire breadths after SARS-CoV-2 infection, too. The reasons why AZ and BNT primary vaccination elicits different immune response patterns to essentially the same antigen, and the associated benefits and risks, need further investigation to inform vaccine and vaccination schedule development

An AHP (Analytic Hierarchy Process)/ANP (Analytic Network Process)-based multi- criteria decision approach for the selection of solar-thermal power plant investment projects

In this paper the AHP (Analytic Hierarchy Process) and the ANP (Analytic Network Process) are applied to help the managing board of an important Spanish solar power investment company to decide whether to invest in a particular solar-thermal power plant project and, if so, to determine the order of priority of the projects in the company's portfolio. Project management goes through a long process, from obtaining the required construction permits and authorizations, negotiating with different stakeholders, complying with complex legal regulations, to solving the technical problems associated with plant construction and distribution of the energy generated. The whole process involves high engineering costs. The decision approach proposed in this paper consists of three phases. In the first two phases, the managing board must decide whether to accept or reject a project according to a set of criteria previously identified by the technical team. The third phase consists of establishing a priority order among the projects that have proven to be economically profitable based on project risk levels and execution time delays. This work analyzes the criteria that should be taken into account to accept or reject proposals for investment, as well as the risks used to prioritize some projects over others.The translation of this paper has been funded by the Universitat Politecnica de Valencia.Aragonés Beltrán, P.; Chaparro González, FV.; Pastor Ferrando, JP.; Pla Rubio, A. (2014). An AHP (Analytic Hierarchy Process)/ANP (Analytic Network Process)-based multi- criteria decision approach for the selection of solar-thermal power plant investment projects. Energy. 66:222-238. doi:10.1016/j.energy.2013.12.016S2222386

Peptide microarray based analysis of antibody responses to SARS-CoV-2 identifies unique epitopes with potential for diagnostic test development

Humoral immunity to the Severe Adult Respiratory Syndrome (SARS) Coronavirus (CoV)‐2 is not fully understood yet but is a crucial factor of immune protection. The possibility of antibody cross‐reactivity between SARS‐CoV‐2 and other human coronaviruses (HCoVs) would have important implications for immune protection but also for the development of specific diagnostic ELISA tests. Using peptide microarrays, n = 24 patient samples and n = 12 control samples were screened for antibodies against the entire SARS‐CoV‐2 proteome as well as the Spike (S), Nucleocapsid (N), VME1 (V), R1ab, and Protein 3a (AP3A) of the HCoV strains SARS, MERS, OC43 and 229E. While widespread cross‐reactivity was revealed across several immunodominant regions of S and N, IgG binding to several SARS‐CoV‐2‐derived peptides provided statistically significant discrimination between COVID‐19 patients and controls. Selected target peptides may serve as capture antigens for future, highly COVID‐19‐specific diagnostic antibody tests

Building ProteomeTools based on a complete synthetic human proteome.

We describe ProteomeTools, a project building molecular and digital tools from the human proteome to facilitate biomedical research. Here we report the generation and multimodal liquid chromatography-tandem mass spectrometry analysis of \u3e330,000 synthetic tryptic peptides representing essentially all canonical human gene products, and we exemplify the utility of these data in several applications. The resource (available at http://www.proteometools.org) will be extended to \u3e1 million peptides, and all data will be shared with the community via ProteomicsDB and ProteomeXchange

Mechanism of Thermal Reversal of the (Fulvalene)tetracarbonyldiruthenium Photoisomerization: Toward Molecular Solar-Thermal Energy Storage

In the currently intensifying quest to harness solar energy for the powering of our planet, most efforts are centered around photoinduced generic charge separation, such as in photovoltaics, water splitting, other small molecule activation, and biologically inspired photosynthetic systems. In contrast, direct collection of heat from sunlight has received much less diversified attention, its bulk devoted to the development of concentrating solar thermal power plants, in which mirrors are used to focus the sun beam on an appropriate heat transfer material. An attractive alternative strategy would be to trap solar energy in the form of chemical bonds, ideally through the photoconversion of a suitable molecule to a higher energy isomer, which, in turn, would release the stored energy by thermal reversal. Such a system would encompass the essential elements of a rechargeable heat battery, with its inherent advantages of storage, transportability, and use on demand. The underlying concept has been explored extensively with organic molecules (such as the norbornadiene-quadricyclane cycle), often in the context of developing photoswitches. On the other hand, organometallic complexes have remained relatively obscure in this capacity, despite a number of advantages, including expanded structural tunability and generally favorable electronic absorption regimes. A highly promising organometallic system is the previously reported, robust photo-thermal fulvalene (Fv) diruthenium couple 1 {l_reversible} 2 (Scheme 1). However, although reversible and moderately efficient, lack of a full, detailed atom-scale understanding of its key conversion and storage mechanisms have limited our ability to improve on its performance or identify optimal variants, such as substituents on the Fv, ligands other than CO, and alternative metals. Here we present a theoretical investigation, in conjunction with corroborating experiments, of the mechanism for the heat releasing step of 2 {yields} 1 and its Fe (4) and Os (6) relatives. The results of the combined study has enabled a rigorous interpretation of earlier and new experimental measurements and paint a surprising picture. First-principles calculations were employed based on spin unrestricted density functional theory (DFT) with a non-empirical gradient corrected exchange-correlation functional. Ultrasoft pseudopotentials were used to describe the valence-core interactions of electrons, including scalar relativistic effects of the core. Wavefunctions and charge densities were expanded in plane waves with kinetic energies up to 25 and 200 Rydberg, respectively. Reaction pathways were delineated with the string method, as implemented within the Car-Parrinello approach. This method allows for the efficient determination of the minimum energy path (MEP) of atomistic transitions and thus also saddle points (transition states, TSs), which are the energy maxima along the MEP. All geometries were optimized until all forces on the atoms were less than 0.02 eV/{angstrom}. The calculated structures of 1 and 2 were in good agreement with their experimental counterparts

Fluorinated Carbohydrates as Lectin Ligands: Biorelevant Sensors with Capacity to Monitor Anomer Affinity in 19F-NMR-Based Inhibitor Screening

11 páginas, 4 figurasd, 2 esquemas -- PAGS nros. 4354-4364Our recent description of pulse sequences for the intramolecular relay of 1H polarization to the 19F reporter of a carbohydrate ligand after saturation transfer from a cognate lectin prompted us to test the applicability of this technique for inhibitor screening. By strategically combining synthetic organic chemistry and cell assays with 19F-NMR-based competition experiments, we document the validity of this approach. Two mannose-specific leguminous agglutinins as receptors, the α-methyl derivative of 2-deoxy-2-fluoro-D-mannopyranoside as sensor and synthetic mannosides selected to represent different inhibitory capacities were used to establish a test panel. Signal amplitudes were found to vary among the two related lectins, and their precipitation by glycodendrimers enabled inherent limits to be set. The obtained experimental basis was then broadened by assaying a galactopyranoside-binding plant toxin, using 6-deoxy-6-fluoro-D-galactopyranose as a sensor molecule. The easy identification of the two anomeric signals by 19F NMR spectroscopy enabled ready detection of the preference of this lectin for the α-anomer and allowed the two individual inhibition profiles to be deduced. These 19F NMR spectroscopic data were in accord with the activities of inhibitors used to protect cells from toxicity. Our results therefore provide an experimental basis for 19F-NMR-based inhibitor screeningThis work was generously supported by the European Community (Marie Curie Research training grant MCRTN-CT-2005–19561), the Ministerio de Ciencia e Innovación (MICINN) (grant numbers CTQ2009-08536 and BFU2009-10052), and the Spanish Institute of Health Carlos III (ISCIII), under the initiative CIBER of Respiratory Diseases (CIBERES). The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement number 260600 (“GlycoHIT”)Peer reviewe

Cross-reactive CD4+ T cells enhance SARS-CoV-2 immune responses upon infection and vaccination

The functional relevance of pre-existing cross-immunity to SARS-CoV-2 is a subject of intense debate. Here, we show that human endemic coronavirus (HCoV)-reactive and SARS-CoV-2-cross-reactive CD4+ T cells are ubiquitous but decrease with age. We identified a universal immunodominant coronavirus-specific spike peptide (S816-830) and demonstrate that pre-existing spike- and S816-830-reactive T cells were recruited into immune responses to SARS-CoV-2 infection and their frequency correlated with anti-SARS-CoV-2-S1-IgG antibodies. Spike-cross-reactive T cells were also activated after primary BNT162b2 COVID-19 mRNA vaccination displaying kinetics similar to secondary immune responses. Our results highlight the functional contribution of pre-existing spike-cross-reactive T cells in SARS-CoV-2 infection and vaccination. Cross-reactive immunity may account for the unexpectedly rapid induction of immunity following primary SARS-CoV-2 immunization and the high rate of asymptomatic/mild COVID-19 disease courses

Solar thermal power plants

The solar thermal power plant technology, the opportunities it presents and the developments in the market are outlined. The focus is on the technology of parabolic trough power plants, a proven technology for solar power generation on a large scale. In a parabolic trough power plant, trough-shaped mirrors concentrate the solar irradiation onto a pipe in the focal line of the collector. The thermal energy thus generated is used for electricity generation in a steam turbine. Parabolic trough plants can be combined with thermal storage and fossil or biomass fired heat exchangers to generate electricity even when the sun is not shining. Solar Millennium AG in Erlangen has developed the first power plant of this kind in Europe. After two years of construction the plant started operation in Southern Spain in 2008. This one and its sister projects are important steps leading the way for the whole market. The paper also covers the technological challenges, the key components used and the research and development activities concerning this technology. Solar thermal power plants are ideal for covering peak and medium loads in power grids. In hybrid operation they can also cover baseload. The Solar Chimney power plant, another striking technology for the conversion of solar into electric energy, is described briefly. The paper concludes with a look at the future – the import of solar energy from the deserts of North Africa to central Europe