Basic Pathways Research Articles

Game Over for Russian Athletes? Human Rights Aspects of Measures Adopted by International Sports Organisations as a Response to the Russian Aggression against Ukraine

Summary The Russian invasion of Ukraine has tested the mettle of the international community, prompting not only States but also non-state actors to take deterrent action in response. Indeed, international sports organisations, namely the Fédération Internationale de Football Association (FIFA) and the Federation of European Football Associations (UEFA), have been relatively successful in shifting the power dynamics by introducing a complete ban on the Football Union of Russia (FUR), i.e., the Russian national and club teams. This article investigates the human rights implications of such actions by international sports organisations. First, the article departs from an assessment of the legal status of FIFA and UEFA under international law. It reflects how a legal link could be established vis-à-vis their human rights obligations. Second, it examines the human rights aspects of the impugned measures by FIFA and UEFA on the part of the Russian athletes, further scrutinising them against the international human rights law principle of non-discrimination through a proportionality test. Finally, it draws basic pathways for how possible human rights violations committed in the context of measures adopted by such organisations could be remedied, outlining the challenges of arbitration and litigation in Switzerland.

Constrained tropical land temperature-precipitation sensitivity reveals decreasing evapotranspiration and faster vegetation greening in CMIP6 projections

Over the tropical land surface, accurate estimates of future changes in temperature, precipitation and evapotranspiration are crucial for ecological sustainability, but remain highly uncertain. Here we develop a series of emergent constraints (ECs) by using historical and future outputs from the Coupled Model Inter-comparison Project Phase 6 (CMIP6) Earth System Models under the four basic Shared Socio-economic Pathway scenarios (SSP126, SSP245, SSP370, and SSP585). Results show that the temperature sensitivity to precipitation during 2015–2100, which varies substantially in the original CMIP6 outputs, becomes systematically negative across SSPs after application of the EC, with absolute values between −1.10 °C mm−1 day and −3.52 °C mm−1 day, and with uncertainties reduced by 9.4% to 41.4%. The trend in tropical land-surface evapotranspiration, which was increasing by 0.292 mm yr−1 in the original CMIP6 model outputs, becomes significantly negative (−0.469 mm yr−1) after applying the constraint. Moreover, we find a significant increase of 58.7% in the leaf area index growth rate.

CCIBP: a comprehensive cosmetic ingredients bioinformatics platform.

Cosmetics form an important part of our daily lives, and it is therefore important to understand the basic physicochemical properties, metabolic pathways, and toxicological and safe concentrations of these cosmetics molecules. Therefore, comprehensive cosmetic ingredients bioinformatics platform (CCIBP) was developed here, which is a unique comprehensive cosmetic database providing information on regulations, physicochemical properties, and human metabolic pathways for cosmetic molecules from major regions of the world, whilst also correlating plant information in natural products. CCIBP supports formulation analysis, efficacy component analysis, and also combines knowledge of synthetic biology to facilitate access to natural molecules and biosynthetic production. CCIBP, empowered with chemoinformatics, bioinformatics, and synthetic biology data and tools, presents a very helpful platform for cosmetic research and development of ingredients. CCIBP is available at: http://design.rxnfinder.org/cosing/.

Minimising Leachate Wastewater Generated from NaOH-Catalysed Biodiesel Synthesis from Methanol

Currently, energy generation based on fossil fuels is producing negative environmental impacts; two of the main symptoms of these impacts are water pollution and climate change. Consequently, the search for new technologies to satisfy the energy demand must have the goal to minimise possible impacts to the environment. There are alternatives with biofuels and, among them, biodiesel. The cheapest reaction pathway for biodiesel production is the transesterification of triglycerides by methanol in the presence of sodium hydroxide; however, this option can contaminate large volumes of water used in the final leach of the biodiesel product. Therefore, a feasible way of producing this biofuel while simultaneously minimising leaching water will be environmentally friendly and will improve economical savings. The present study developed an experimental design in order to minimise the addition of NaOH during biodiesel production by the basic homogeneous pathway. The best operating conditions were 46 °C, methanol in situ 7.5% v/v and NaOH 0.035 M. These conditions allowed to reduce the leaching water amount by 25% compared to techniques reported in the literature; however, the yield to biodiesel decreased from 98 wt.% to 87 wt.% when a model waste oil was used instead of virgin oil.

An atomic insight into reaction pathways and temperature effects in the degradation of polyethylene, polypropylene and polystyrene

Increasingly severe environmental pollution has spurred widespread concern regarding thermoplastics degradation. However, a lack of better understanding at the atomic level hinders the regulation of the related degradation processes. Through theoretical studies of three different typical plastics, first-principles calculations showed two basic reaction pathways for degradation: depolymerization and hydrogen-transfer-induced chain scission. The analyses of the reaction rates were consistent with those of the degradation energy barriers. Even at high temperatures, the differences in depolymerization and hydrogen transfer reactions in reaction rates and thermal perturbation probabilities are still greater than three orders of magnitude. Moreover, dynamic simulations confirm the existence of two reaction pathways and demonstrate that the two pathways occur sequentially. Our findings enrich the understanding of the basic processes of plastic degradation and may provide a reference for pyrolytic processing in waste management.

Characterization of Schistosome Sox Genes and Identification of a Flatworm Class of Sox Regulators.

Schistosome helminths infect over 200 million people across 78 countries and are responsible for nearly 300,000 deaths annually. However, our understanding of basic genetic pathways crucial for schistosome development is limited. The sex determining region Y-box 2 (Sox2) protein is a Sox B type transcriptional activator that is expressed prior to blastulation in mammals and is necessary for embryogenesis. Sox expression is associated with pluripotency and stem cells, neuronal differentiation, gut development, and cancer. Schistosomes express a Sox-like gene expressed in the schistosomula after infecting a mammalian host when schistosomes have about 900 cells. Here, we characterized and named this Sox-like gene SmSOXS1. SmSoxS1 protein is a developmentally regulated activator that localizes to the anterior and posterior ends of the schistosomula and binds to Sox-specific DNA elements. In addition to SmSoxS1, we have also identified an additional six Sox genes in schistosomes, two Sox B, one SoxC, and three Sox genes that may establish a flatworm-specific class of Sox genes with planarians. These data identify novel Sox genes in schistosomes to expand the potential functional roles for Sox2 and may provide interesting insights into early multicellular development of flatworms.

Nutrient Therapy for the Improvement of Fatigue Symptoms

Fatigue, characterised by lack of energy, mental exhaustion and poor muscle endurance which do not recover following a period of rest, is a common characteristic symptom of several conditions and negatively impacts the quality of life of those affected. Fatigue is often a symptom of concern for people suffering from conditions such as fibromyalgia, chronic fatigue syndrome, cancer, and multiple sclerosis. Vitamins and minerals, playing essential roles in a variety of basic metabolic pathways that support fundamental cellular functions, may be important in mitigating physical and mental fatigue. Several studies have examined the potential benefits of nutrients on fatigue in various populations. The current review aimed to gather the existing literature exploring different nutrients’ effects on fatigue. From the searches of the literature conducted in PubMed, Ovid, Web of Science, and Google scholar, 60 articles met the inclusion criteria and were included in the review. Among the included studies, 50 showed significant beneficial effects (p < 0.05) of vitamin and mineral supplementation on fatigue. Altogether, the included studies investigated oral or parenteral administration of nutrients including Coenzyme Q10, L-carnitine, zinc, methionine, nicotinamide adenine dinucleotide (NAD), and vitamins C, D and B. In conclusion, the results of the literature review suggest that these nutrients have potentially significant benefits in reducing fatigue in healthy individuals as well as those with chronic illness, both when taken orally and parenterally. Further studies should explore these novel therapies, both as adjunctive treatments and as sole interventions.

Investigation on the mercury nucleation mechanism based on the release characteristics of inhalable particles during coupled combustion of sewage sludge-coal washery tailing

Establishing an integrated treatment technology system for sewage sludge and coal washery tailing by mixed combustion can effectively overcome the disadvantages of the low calorific value and utilization efficiency, realizing the efficient resource. The single-component and coupled fuel were analyzed. Aiming at the generated inhalable particulate matter during the combustion process, using multiple characterization means and the fluidized bed experimental system, the micro-characteristics and elemental distribution of coupled fuel and combustion products were obtained, and the theory based on nucleation control was analyzed to reveal particle size distribution and particulate matter emission characteristics under multiparameter coupling conditions. The nucleation mechanism of Hg on the surface of the particulate matter under mixed combustion conditions was clarified. The results show that the weight loss process of sewage sludge in the combustion process mainly includes four stages. An obvious boundary between the volatilization and combustion processes of fixed carbon does not exist; furthermore, there is some overlap between the two processes. Volatiles and minerals interact in the mixed combustion process. The constituent elements of PM1 and PM1∼10 in the generated particulate matter are mainly easily vaporizable and refractory elements, respectively. The particle size of the particles produced presents the characteristics of bimodal distribution. There is a critical value (800 °C) for the influence of combustion temperature on the emission characteristics of particulate matter. It mainly affects the emission characteristics of coarse and fine particles through dominating the occurrence forms of tar, easily gasified and refractory elements, and the collision and aggregation process between particles. Homogeneous nucleation and heterogeneous condensation are the basic pathways for the transition of Hg from the gas phase to particle nucleation, and there are different nucleation control mechanisms. The heterogeneous condensation process includes diffusion control and surface reaction control. In addition, after the coupled fuel is demineralized by acid washing, the nucleation control mechanism changes significantly from diffusion control to chemical reaction control for the particles in the continuum region.

Complosome - the intracellular complement system.

The complement system is a recognized pillar of host defence against infection and noxious self-derived antigens. Complement is traditionally known as a serum-effective system, whereby the liver expresses and secretes most complement components, which participate in the detection of bloodborne pathogens and drive an inflammatory reactionto safely remove the microbial or antigenic threat. However, perturbations in normal complement function can cause severe disease and, for reasons that are currently not fully understood, the kidney isparticularly vulnerable to dysregulated complement activity. Novel insights into complement biology have identified cell-autonomous and intracellularly active complement - the complosome - as an unexpected central orchestrator of normal cell physiology. For example, thecomplosome controls mitochondrial activity, glycolysis, oxidative phosphorylation, cell survival and gene regulation in innate and adaptive immune cells, and in non-immune cells, such as fibroblasts and endothelial and epithelial cells. These unanticipated complosome contributions to basic cell physiological pathways make it a novel and central player in the control of cell homeostasis and effector responses. This discovery, together with the realization that an increasing number of human diseases involve complement perturbations, has renewed interest in the complement system and its therapeutic targeting. Here, we summarize the current knowledge about the complosome across healthy cells and tissues, highlight contributions from dysregulated complosome activities to human disease and discuss potential therapeutic implications.

Stability-indicating HPLC-DAD and TLC-densitometry methods for the quantification of bupivacaine and meloxicam in their co-formulated mixture

The implementation of a novel drug formulation necessitates the assessment of its content stability and the designation of appropriate stability monitoring methodologies. Bupivacaine (BVC) and Meloxicam (MLX) were recently authorized under the name Zynrelef® solution. This novel combination was tested for stability under a variety of stress circumstances for the first time in this study. The basic degradation pathway was shown to affect BVC, while the acidic degradation pathway works for MLX. Identification of the acquired degradation products, including BVC impurity F (BVC IMP-F), 1-buytylpiperidine-2- 2carboxylic acid (1-BPC), MLX acid degradation product (MLX-DEG), and MLX impurity B (MLX IMP-B), was carried out using IR and MS spectroscopic techniques. HPTLC and HPLC, two chromatographic-based stability-indicating methods, have been established. The HPTLC method offered efficient separation upon the usage of silica gel 60 F254 as a stationary phase and ethyl acetate/toluene/triethylamine/acetic acid (4:4:2:0.4, v/v/v/v) as a developing system with densitometric recording at 260 nm. The HPLC method was developed by using Zorbax® SB C18 column as a stationary phase and a mobile phase consisting of acetonitrile: methanol: phosphate buffer, pH 6.5 (20:60:20 v/v/v), at a flow rate of 1.5 mL min-1. Quantification of the analytes was performed at 265 nm using a diode array detector. The current research introduced the first stability-indicating methods for the assay of BVC and MLX which were successfully applied for the determination of the cited drugs in their combined pharmaceutical formulation. Moreover, the HPLC method was further extended for studying the degradation kinetics of the two drugs.

Re-thinking the role of exploratory scenarios for adaptive policymaking: An application for land use and transport planning

While scenario-building has proven useful in situations marked by considerable uncertainty in the context of land use and transport policymaking, additional research is needed to develop adaptive and dynamic scenario-building schemes. To address this gap, the present paper conceptualises an adaptive scenario-building approach that would facilitate adaptive policy responses by using exploratory scenarios as intermediate tipping points. An empirical research design is also conducted following a participatory approach, with a case study of the Henares Corridor in the Metropolitan Area of Madrid, Spain. Three exploratory scenarios are assessed as experimental tipping points for the basic policy pathways initially adopted by regional and local governments. The use of such tipping points suggested different adaptive policy responses: (i) preserving policy responses when exploratory scenarios are strongly aligned with the basic pathways; and (ii) transforming policy responses when significant deviations are perceived between the exploratory scenarios and the basic pathways. The paper also discusses the real-life application potential of this adaptive scenario-building approach, further research steps, and future horizons for scenario-building.

Advances in Electricity-Steering Organic Waste Bio-Valorization for Medium Chain Carboxylic Acids Production

Medium chain carboxylic acids (MCCAs, e.g., caproic acid, caprylic acid, etc.) with 6–12 carbon atoms are valuable platform chemicals produced from organic waste via microbial chain elongation metabolism named as reversed β-oxidation and fatty acid-biosynthesis cyclical pathway. Recently, many articles reported that electricity could not only serve as the external electron donor and provide the reduction equivalent required for chain elongation but also regulate the microbiome structure and metabolic behaviors to promote MCCAs formation. Electricity-steering MCCAs bioproduction has become an appealing technique to valorize low-value organic waste, paving an alternative pathway for net-zero carbon emission energy systems and sustainable socio-economic development. However, the MCCAs’ bioproduction from organic waste steered by electric field has not been comprehensively reviewed. From a systematical analysis of publicly available literature, we first covered the basic working principle, fermentation architecture, functional microflora, and metabolic pathway of MCCAs production driven by electricity. The strategies of substrate modulation, applied voltage/current regulation, electrode optimization, and microbial cooperation and stimulation for boosting electricity-driven MCCAs bioproduction are then scrutinized and extensively discussed. Ultimately, the pressing knowledge gaps and the potential path forward are proposed to provide pointers for consistently higher MCCAs yield and the transition from laboratory to market.

A Novel Glycoside Hydrolase DogH Utilizing Soluble Starch to Maltose Improve Osmotic Tolerance in Deinococcus radiodurans.

Deinococcus radiodurans is a microorganism that can adjust, survive or thrive in hostile conditions and has been described as "the strongest microorganism in the world". The underlying mechanism behind the exceptional resistance of this robust bacterium still remains unclear. Osmotic stress, caused by abiotic stresses such as desiccation, salt stress, high temperatures and freezing, is one of the main stresses suffered by microorganisms, and it is also the basic response pathway by which organisms cope with environmental stress. In this study, a unique trehalose synthesis-related gene, dogH (Deinococcus radiodurans orphan glycosyl hydrolase-like family 10), which encodes a novel glycoside hydrolase, was excavated using a multi-omics combination method. The content accumulation of trehalose and its precursors under hypertonic conditions was quantified by HPLC-MS. Ours results showed that the dogH gene was strongly induced by sorbitol and desiccation stress in D. radiodurans. DogH glycoside hydrolase hydrolyzes α-1,4-glycosidic bonds by releasing maltose from starch in the regulation of soluble sugars, thereby increasing the concentration of TreS (trehalose synthase) pathway precursors and trehalose biomass. The maltose and alginate content in D. radiodurans amounted to 48 μg mg protein-1 and 45 μg mg protein-1, respectively, which were 9 and 28 times higher than those in E. coli, respectively. The accumulation of greater intracellular concentrations of osmoprotectants may be the true reason for the higher osmotic stress tolerance of D. radiodurans.

Dysfunctional Lipid Metabolism-The Basis for How Genetic Abnormalities Express the Phenotype of Aggressive Prostate Cancer.

Prostate cancer is the second most frequent cancer in men, with increasing prevalence due to an ageing population. Advanced prostate cancer is diagnosed in up to 20% of patients, and, therefore, it is important to understand evolving mechanisms of progression. Significant morbidity and mortality can occur in advanced prostate cancer where treatment options are intrinsically related to lipid metabolism. Dysfunctional lipid metabolism has long been known to have a relationship to prostate cancer development; however, only recently have studies attempted to elucidate the exact mechanism relating genetic abnormalities and lipid metabolic pathways. Contemporary research has established the pathways leading to prostate cancer development, including dysregulated lipid metabolism-associated de novo lipogenesis through steroid hormone biogenesis and β-oxidation of fatty acids. These pathways, in relation to treatment, have formed potential novel targets for management of advanced prostate cancer via androgen deprivation. We review basic lipid metabolism pathways and their relation to hypogonadism, and further explore prostate cancer development with a cellular emphasis.

Gossypol and related compounds are produced and accumulate in the aboveground parts of the cotton plant, independent of roots as the source.

Use of Ultra-low gossypol cottonseed event as a scion in a graft combination confirmed that roots are not a source of terpenoids in the aboveground parts of a cotton plant. Gossypol and related terpenoids, derived from the same basic biosynthetic pathway, are present in the numerous lysigenous glands in the aboveground parts of a cotton plant. Roots, with sparse presence of such glands, do produce significant amount of gossypol and a different set of terpenoids. These compounds serve a defensive function against various pests and pathogens. This investigation was undertaken to examine whether gossypol produced in the roots can replenish the gossypol content of the cottonseed-glands that are largely devoid of this terpenoid in a genetically engineered event. Graft unions between a scion derived from the RNAi-based, Ultra-low gossypol cottonseed (ULGCS) event, TAM66274, and a rootstock derived from wild-type parental genotype, Coker 312 (Coker), were compared with various other grafts that served as controls. The results showed that the seeds developing within the scion of test grafts (ULGCS/Coker) continued to maintain the ultra-low gossypol levels found in the TAM66274 seeds. Molecular analyses confirmed that while the key gene involved in gland development showed normal activity in the developing embryos in the scion, two genes encoding the enzymes involved in gossypol biosynthesis were suppressed. Thus, the gene expression data confirmed the results obtained from biochemical measurements and collectively demonstrated that roots are not a source of gossypol for the aboveground parts of the cotton plant. These findings, combined with the results from previous investigations, support the assertion that gossypol and related terpenoids are produced in a highly localized manner in various organs of the cotton plant and are retained therein.

Accumulation of pyrethroids induces changes in metabolism of the entomopathogenic fungus Beauveria bassiana—Proteomic and lipidomic background

Advances in the agrochemical industry, such as using plant protection products e.g. pyrethroid insecticides, lead to environmental pollution via the accumulation of toxic compounds in soil. An interesting approach to overcoming this threat is using biopreparations based on entomopathogenic fungi that come into contact with the residues of the insecticides in the environment. The aim of this study was to determine whether the soil-dwelling entomopathogenic fungus Beauveria bassiana ARSEF 2860 is capable of accumulating pyrethroids (λ-cyhalothrin, α-cypermethrin and deltamethrin) and to identify the metabolomics and proteomic implications of this process. In this work, we demonstrated for the first time that the tested fungus accumulated pyrethroids as early as on day 2 of incubation with an average efficiency of 90%. Pyrethroids accumulated in large quantities in the mycelium of B. bassiana induced oxidative stress and interacted differently with the enzymes of the basic metabolic pathways, enzymes associated with the organization of the actin cytoskeleton and cell walls, as well as extracellular enzymes responsible for the infectious abilities (α-cypermethrin caused a 61% decrease in PR1, λ-cyhalothrin - a 31% decrease in PR2, which are proteolytic enzymes with a confirmed role in the infectious process). This study also revealed that the accumulated pyrethroids decreased the activity of phospholipase C, which increased the triacylglycerols/diacylglycerols (TAG/DAG) ratio, especially in mycelium in which α-cypermethrin was accumulated. It should be emphasized that the accumulation of pyrethroids in the environment is not fully understood, and current research suggests that entomopathogenic fungi may be part of the process.

Incorporation of tissue factor-integrated liposome and silica nanoparticle into collagen hydrogel as a promising hemostatic system

Bleeding complications are associated with substantial tissue morbidities and mortalities. Biomimetic composite materials that possess the ability to sufficiently stimulate and augment different physiological mechanisms of hemostasis are highly desirable to reduce bleeding-related casualties, which, however, are still largely under-explored. This study aims to develop a composite hemostatic system by combining collagen hydrogel with tissue factor (TF)-integrated liposome and silica nanoparticle, which could integrate the platelet plug-promoting capacity of collagen with the abilities of the latter two components to activate the extrinsic and intrinsic pathways of coagulation respectively. Several hydrogel compositions were synthesized and characterized. We show that lipidated TF and silica were evenly distributed in the collagen-based hydrogels, while exhibiting tunable release kinetics in simulated body fluid. Time-to-coagulation test revealed that each component in the TF-liposome/silica/collagen ternary hydrogels was hemostasis-active, and their combination showed enhanced and potent procoagulant performance, without detectable cytotoxicity against NIH/3T3 model cells. These results suggest that collagen hydrogels with embedded TF-liposome and silica nanoparticle may serve as a platform for an effective hemostatic composite that incorporates all the basic known pathways of coagulation.

A molecular network map of orexin-orexin receptor signaling system.

Orexins are excitatory neuropeptides, which are predominantly associated with feeding behavior, sleep-wake cycle and energy homeostasis. The orexinergic system comprises of HCRTR1 and HCRTR2, G-protein-coupled receptors of rhodopsin family and the endogenous ligands processed from HCRT pro-hormone, Orexin A and Orexin B. These neuropeptides are biosynthesized by the orexin neurons present in the lateral hypothalamus area, with dense projections to other brain regions. The orexin-receptor signaling is implicated in various metabolic as well as neurological disorders, making it a promising target for pharmacological interventions. However, there is limited information available on the collective representation of the signal transduction pathways pertaining to the orexin-orexin receptor signaling system. Here, we depict a compendium of the Orexin A/B stimulated reactions in the form of a basic signaling pathway map. This map catalogs the reactions into five categories: molecular association, activation/inhibition, catalysis, transport, and gene regulation. A total of 318 downstream molecules were annotated adhering to the guidelines of NetPath curation. This pathway map can be utilized for further assessment of signaling events associated with orexin-mediated physiological functions and is freely available on WikiPathways, an open-source pathway database ( https://www.wikipathways.org/index.php/Pathway:WP5094 ).

Metabolic Alterations and Elevation of Oxidative Stress during the Early Post-Transplant Period in AML Patients

Introduction Metabolic reprogramming as one of the hallmarks of cancer is necessary to satisfy the increased cell demands for energy and biosynthetic precursors. Our earlier study showed that the concentrations of TCA cycle, glycolysis metabolites, and oxidative stress markers differ significantly among individual stages of AML patient treatment before transplantation, but do not reach the values of healthy metabolism. In this work, we focused on pre-transplant (conditioning regimens) and early post-transplant periods in AML patients, specifically we monitored changes in concentrations of proteinogenic amino acids, metabolites of basic metabolic pathways, and oxidative stress markers. The aim of this study was to characterize concentration profiles of selected metabolites and oxidative stress markers during pre-transplant and early post-transplant periods and to reveal their possible influence on post-transplantation development. Methods Metabolomic profiles were generated using serum samples obtained at pre- and post-transplantation from selected patients treated in the Institute of Hematology and Blood Transfusion, Prague, Czech Republic. Targeted metabolomic profiling of selected metabolites related mainly to TCA cycle, glycolysis, proteinogenic amino acid, and markers of oxidative stress was performed by LC-MS/MS. Results Using non-parametric Wilcoxon test we found a significant decrease in most of the quantified metabolites of the citrate cycle and glycolysis during conditioning regimens. Conversely, in the case of proteinogenic amino acids, we found significant changes in 14 proteinogenic amino acids, specifically in 12 of them there was a significant increase during the conditioning regimens. To assess oxidative stress, we quantified total malondialdehyde as a marker of lipoperoxidation and pyroglutamate as a marker of glutathione depletion. We found that during conditioning regimens, there was a significant increase in malondialdehyde and a simultaneous increase in pyroglutamate. Moreover, we found that oxidative stress presented by malondialdehyde concentration significantly increased 7, 14, and 21 days after transplantation in comparison to the state before conditioning regimens. The lowest decrease in concentrations of TCA cycle and glycolysis metabolites occurred around day 7 post-transplantation. The suppression of monitored metabolic pathways resolved after 21 days post-transplantation. Conclusion The results of our preliminary targeted metabolomic profiling of AML patients contribute to expanding the knowledge of metabolite concentrations and their changes during pre/post-transplantation. Together with the description of the level of oxidative stress, it may reveal a possible influence on development after transplantation and could have a potential impact on supportive treatment regimens even after transplantation. Acknowledgement This work was supported by the European Regional Development Fund and the state budget of the Czech Republic (project AIIHHP: CZ.02.1.01/0.0/0.0/16_025/0007428, OP RDE, Ministry of Education, Youth and Sports) and by the project of the Ministry of Health, Czech Republic (00023736).

DDDR-18. COMPREHENSIVE PROTEOMIC PROFILING OF BRAFV600E GLIOMA CELLS SHOWS UPREGULATION OF GLYCOLYSIS AND TCA CYCLE PROTEINS AFTER COMBINED MTOR AND MAP KINASE PATHWAY BLOCKADE

Abstract Low-grade gliomas (LGG) are slow-growing tumors that affect neurologic function by pressing on the surrounding parts of the brain. Pediatric low-grade glioma (pLGG) is the most common childhood brain tumors. Patients with pLGG often have a significant life-long disabilities. The most common mutations in pLGG are in the mitogen-activated protein kinase (MAPK) pathway including tumor suppressor neurofibromin (NF1), and the oncogene BRAF. BRAF activates MEK1/2, which activates of ERK1/2, leading to tumorigenesis. BRAFV600E mutation results in the activation segment of the kinase “monomeric form of BRAF”, leading to constitutive activation of MAP kinase and mTOR pathways Combination treatment of TAK228 (mTOR inhibitor) and trametinib (MEK1/2 inhibitor), suppressed both mTOR and MAP kinase pathways and inhibited BRAFV600E glioma tumor growth. To further investigate and understand the molecular mechanism of TAK228 and trametinib combinatorial efficacy, we performed comprehensive proteomic analysis of BRAFV600E glioma tumors treated with TAK228 and trametinib. We found increased levels of many proteins in the glycolysis and TCA cycle. Specifically we found increases at the protein level in citrate synthase, aconitase and malate dehydrogenase in the TCA cycle. In glycolysis, we observed increased glucose transporter expression (SLC2A1), increased hexokinase 2, and increased GAPDH. Upregualtion of these proteins may represent attempts to bypass the effects of mTOR and MAP kinase blockade on the activity of these pathways. We are currently performing flux metabolic profiling using uniformly labeled glucose to assess the activity of the pathways after combined mTOR and MAP kinase blockade. Our results may increase our understanding of how glioma cells use glucose and how mTOR and MAP kinase pathway inhibition affects basic metabolic pathways in glioma cells.