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At least 1,135 records · Page 63

Solid State Ionics - the key to the discovery, introduction and domination of lithium batteries for portable energy storage

2017 is the 50th anniversary of the discovery of fast ion transport in β-alumina. This discovery led to a revolution in the study of solid materials that might exhibit ionic motion as fast as that found in liquids. The need for materials with both fast ionic and electronic conductivity and a wide stoichiometry range to measure the ionic conductivity of beta alumina led to the intercalation concept for rechargeable alkali metal batteries. These discoveries and the resulting development of the lithium battery are described in this paper.

25 ENERGY STORAGE↗

Structure of the Mitochondrial Aminolevulinic Acid Synthase, a Key Heme Biosynthetic Enzyme

5-Aminolevulinic acid synthase (ALAS) catalyzes the first step in heme biosynthesis. We present herein the crystal structure of a eukaryotic ALAS from Saccharomyces cerevisiae . In this homodimeric structure, one ALAS subunit contains covalently bound cofactor, pyridoxal 5'-phosphate (PLP), whereas the second is PLP free. Comparison between the subunits reveals PLP-coupled reordering of the active site and of additional regions to achieve the active conformation of the enzyme. The eukaryotic C-terminal extension, a region altered in multiple human disease alleles, wraps around the dimer and contacts active-site-proximal residues. Mutational analysis demonstrates that this C-terminal region that engages the active site is important for ALAS activity. Our discovery of structural elements that change conformation upon PLP binding and of direct contact between the C-terminal extension and the active site thus provides a structural basis for investigation of disruptions in the first step of heme biosynthesis and resulting human disorders.

59 BASIC BIOLOGICAL SCIENCES↗

Simulation and fitting of complex reaction network TPR: The key is the objective function

In this research, a method has been developed for finding improved fits during simulation and fitting of data from complex reaction network temperature programmed reactions (CRN-TPR). It was found that simulation and fitting of CRN-TPR presents additional challenges relative to simulation and fitting of simpler TPR systems. The method used here can enable checking the plausibility of proposed chemical mechanisms and kinetic models. The most important finding was that when choosing an objective function, use of an objective function that is based on integrated production provides more utility in finding improved fits when compared to an objective function based on the rate of production. The response surface produced by using the integrated production is monotonic, suppresses effects from experimental noise, requires fewer points to capture the response behavior, and can be simulated numerically with smaller errors. For CRN-TPR, there is increased importance (relative to simple reaction network TPR) in resolving of peaks prior to fitting, as well as from weighting of experimental data points. Using an implicit ordinary differential equation solver was found to be inadequate for simulating CRN-TPR. Lastly, the method employed here was capable of attaining improved fits in simulation and fitting of CRN-TPR when starting with a postulated mechanism and physically realistic initial guesses for the kinetic parameters.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Differential DNA methylation profile of key genes in malignant prostate epithelial cells transformed by inorganic arsenic or cadmium

Previous work shows altered methylation patterns in inorganic arsenic (iAs)- or cadmium (Cd)-transformed epithelial cells. Here, the methylation status near the transcriptional start site was assessed in the normal human prostate epithelial cell line (RWPE-1) that was malignantly transformed by 10 μM Cd for 11 weeks (CTPE) or 5 μM iAs for 29 weeks (CAsE-PE), at which time cells showed multiple markers of acquired cancer phenotype. Next generation sequencing of the transcriptome of CAsE-PE cells identified multiple dysregulated genes. Of the most highly dysregulated genes, five genes that can be relevant to the carcinogenic process (S100P, HYAL1, NTM, NES, ALDH1A1) were chosen for an in-depth analysis of the DNA methylation profile. DNA was isolated, bisulfite converted, and combined bisulfite restriction analysis was used to identify differentially methylated CpG sites, which was confirmed with bisulfite sequencing. Four of the five genes showed differential methylation in transformants relative to control cells that was inversely related to altered gene expression. Increased expression of HYAL1 (> 25-fold) and S100P (> 40-fold) in transformants was correlated with hypomethylation near the transcriptional start site. Decreased expression of NES (> 15-fold) and NTM (> 1000-fold) in transformants was correlated with hypermethylation near the transcriptional start site. ALDH1A1 expression was differentially expressed in transformed cells but was not differentially methylated relative to control. In conclusion, altered gene expression observed in Cd and iAs transformed cells may result from altered DNA methylation status. - Highlights: • Cd and iAs are known human carcinogens, yet neither appears directly mutagenic. • Prior data suggest epigenetic modification plays a role in Cd or iAs induced cancer. • Altered methylation of four misregulated genes was found in Cd or iAs transformants. • The resulting altered gene expression may be relevant to cellular transformation.

60 APPLIED LIFE SCIENCES↗

Identification of protein expression alterations in gefitinib-resistant human lung adenocarcinoma: PCNT and mPR play key roles in the development of gefitinib-associated resistance

Gefitinib is the first-line chemotherapeutic drug for treating non-small cell lung cancer (NSCLC), which comprises nearly 85% of all lung cancer cases worldwide. However, most patients eventually develop drug resistance after 12–18 months of treatment. Hence, investigating the drug resistance mechanism and resistance-associated biomarkers is necessary. Two lung adenocarcinoma cell lines, PC9 and gefitinib-resistant PC9/Gef, were established for examining resistance mechanisms and identifying potential therapeutic targets. Two-dimensional differential gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry were used for examining global protein expression changes between PC9 and PC9/Gef. The results revealed that 164 identified proteins were associated with the formation of gefitinib resistance in PC9 cells. Additional studies using RNA interference showed that progesterone receptor membrane component 1 and pericentrin proteins have major roles in gefitinib resistance. In conclusion, the proteomic approach enabled identifying of numerous proteins involved in gefitinib resistance. The results provide useful diagnostic markers and therapeutic candidates for treating gefitinib-resistant NSCLC. - Highlights: • 164 proteins associated with gefitinib resistance were identified through proteomic analysis. • In this study, a lung adenocarcinoma and its gefitinib resistant partner were established. • mPR and PCNT proteins have evidenced to play important roles in gefitinib resistance.

60 APPLIED LIFE SCIENCES↗

Identification of the key pathway of oxazolinoanthracyclines mechanism of action in cells derived from human solid tumors

Oxazolinodoxorubicin (O-DOX) and oxazolinodaunorubicin (O-DAU) are novel anthracycline derivatives with a modified daunosamine moiety. In the present study, we evaluated the cytotoxicities, genotoxicities and abilities of O-DOX and O-DAU to induce apoptosis in cancer cell lines (SKOV-3; A549; HepG2), and compared the results with their parent drugs. We assessed antiproliferative activity by MTT assay. We evaluated apoptosis-inducing ability by double-staining with fluorescent probes (Hoechst 33258/propidium iodide), and by determining expression levels of genes involved in programmed cell death by reverse transcription-polymerase chain reaction. Genotoxicities of the compounds were tested by comet assays. Oxazolinoanthracyclines demonstrated high anti-tumor activity. O-DOX had significantly higher cytotoxicity, apoptosis-inducing ability, and genotoxicity compared with parental doxorubicin (DOX) in all tested conditions, while O-DAU activity differed among cell lines. The mechanism of oxazoline analog action appeared to involve the mitochondrial pathway of programmed cell death. These results provide further information about oxazoline derivatives of commonly used anthracycline chemotherapy agents. O-DOX and O-DAU have the ability to induce apoptosis in tumor cells. - Highlights: • Substituted amino group increased the anticancer activity of anthracyclines. • Mitochondrial apoptotic pathway seems to be involved in the mechanism of action. • Favorable biological properties of oxazoline derivatives were confirmed.

60 APPLIED LIFE SCIENCES↗

Key Issues Hindering a Practical Lithium-Metal Anode

The sluggish progress of battery technologies has drastically hindered the rapid development of electric vehicles and next-generation portable electronics. The lithium (Li) metal anode is critical to break the energy-density bottleneck of current Li-ion chemistry. After being intensively studied in recent years, the Li-metal field has developed new understanding and made unprecedented progress in preventing Li-dendrite growth and improving Coulombic efficiency, especially through development of advanced electrolytes and novel analytical tools. Here, we revisit the controversial issues surrounding Li metal as an anode based upon recent advances, revealing the underlying cause of Li metal failure and the true role of ‘solid electrolyte interphase’ in Li-metal anodes. Finally, we propose future directions that must be taken in order for Li-metal batteries to become commercially viable.

25 ENERGY STORAGE↗

Kelvin-Helmholtz instability as one of the key features for fast and efficient emulsification by hydrodynamic cavitation

The paper investigates the oil–water emulsification process inside a micro-venturi channel. More specifically, the possible influence of Kelvin-Helmholtz instability on the emulsification process. High-speed visualizations were conducted inside a square venturi constriction with throat dimensions of 450 µm by 450 µm, both under visible light and X-Rays. We show that cavity shedding caused by the instability results in the formation of several cavity vortices. Their rotation causes the deformation of the oil stream into a distinct wave-like shape, combined with fragmentation into larger drops due to cavitation bubble collapse. Later on, the cavity collapse further disperses the larger drops into a finer emulsion. Thus, it turns out that the Kelvin-Helmholtz instability is similarly characteristic for hydrodynamic cavitation emulsification inside a microchannel as is the Rayleigh-Taylor instability for acoustically driven emulsion formation.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

The Lyssavirus glycoprotein: A key to cross-immunity

Rabies is an acute viral encephalomyelitis in warm-blooded vertebrates, caused by viruses belonging to Rhabdovirus family and genus Lyssavirus. Although rabies is categorised as a neglected disease, the rabies virus (RABV) is the most studied amongst Lyssaviruses which show nearly identical infection patterns. In efforts to improving post-exposure prophylaxis, several anti-rabies monoclonal antibodies (mAbs) targeting the glycoprotein (G protein) sites I, II, III and G5 have been characterized. To explore cross-neutralization capacity of available mAbs and discover new possible B-cell epitopes, we have analyzed all available glycoprotein sequences from Lyssaviruses with a focus on sequence variation and conservation. This information was mapped on the structure of a representative G protein. We proposed several possible cross-neutralizing B-cell epitopes (GUVTTTF, WLRTV, REECLD and EHLVVEEL) in complement to the already well-characterized antigenic sites. The research could facilitate development of novel cross-reactive mAbs against RABV and even more broad, against possibly all Lyssavirus members. -- Highlights: •The current PEP has raised safety and availability concerns. •Cocktails of mAbs have been proposed as alternative treatment. •Amino acid conservation amongst Lyssavirus G proteins was studied. •Possible cross-neutralizing B-cell epitopes were proposed.

60 APPLIED LIFE SCIENCES↗

Assessing recycling versus incineration of key materials in municipal waste: The importance of efficient energy recovery and transport distances

Highlights: Black-Right-Pointing-Pointer We model the environmental impact of recycling and incineration of household waste. Black-Right-Pointing-Pointer Recycling of paper, glass, steel and aluminium is better than incineration. Black-Right-Pointing-Pointer Recycling and incineration of cardboard and plastic can be equally good alternatives. Black-Right-Pointing-Pointer Recyclables can be transported long distances and still have environmental benefits. Black-Right-Pointing-Pointer Paper has a higher environmental benefit than recyclables found in smaller amounts. - Abstract: Recycling of materials from municipal solid waste is commonly considered to be superior to any other waste treatment alternative. For the material fractions with a significant energy content this might not be the case if the treatment alternative is a waste-to-energy plant with high energy recovery rates. The environmental impacts from recycling and from incineration of six material fractions in household waste have been compared through life cycle assessment assuming high-performance technologies for material recycling as well as for waste incineration. The results showed that there are environmental benefits when recycling paper, glass, steel and aluminium instead of incinerating it. For cardboard and plastic the results were more unclear, depending on the level of energy recovery at the incineration plant, the system boundaries chosen and which impact category was in focus. Further, the environmental impact potentials from collection, pre-treatment and transport was compared to the environmental benefit from recycling and this showed that with the right means of transport, recyclables can in most cases be transported long distances. However, the results also showed that recycling of some of the material fractions can only contribute marginally in improving the overall waste management system taking into consideration their limited content in average Danish household waste.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Waste to energy – key element for sustainable waste management

Highlights: • First paper on the importance of incineration from a urban metabolism point of view. • Proves that incineration is necessary for sustainable waste management. • Historical and technical overview of 100 years development of MSW incineration. - Abstract: Human activities inevitably result in wastes. The higher the material turnover, and the more complex and divers the materials produced, the more challenging it is for waste management to reach the goals of “protection of men and environment” and “resource conservation”. Waste incineration, introduced originally for volume reduction and hygienic reasons, went through a long and intense development. Together with prevention and recycling measures, waste to energy (WTE) facilities contribute significantly to reaching the goals of waste management. Sophisticated air pollution control (APC) devices ensure that emissions are environmentally safe. Incinerators are crucial and unique for the complete destruction of hazardous organic materials, to reduce risks due to pathogenic microorganisms and viruses, and for concentrating valuable as well as toxic metals in certain fractions. Bottom ash and APC residues have become new sources of secondary metals, hence incineration has become a materials recycling facility, too. WTE plants are supporting decisions about waste and environmental management: They can routinely and cost effectively supply information about chemical waste composition as well as about the ratio of biogenic to fossil carbon in MSW and off-gas.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Effects of size and thermophilic pre-hydrolysis of banana peel during anaerobic digestion, and biomethanation potential of key tropical fruit wastes

Highlights: • Digestibility of tropical fruit wastes is in order of banana, longan and rambutan. •Size reduction increased CH{sub 4} yield of LF banana peel by 23.2% in batch assay. • H{sub 2} and CH{sub 4} were produced by adding the front-end thermo pre-hydrolysis tank. • Thermo pre-hydrolysis tank provided methanogenic system stability at high solid feed. • Two stage digestion gave higher VS destruction and energy yield at high solid feed. - Abstract: Methane production potential of tropical fruit wastes, namely lady-finger banana peel, rambutan waste and longan waste were compared using BMP assay and stoichiometric modified Buswell and Mueller equation. Methane yields based on volatile solid (VS) were in the order of ground banana peel, chopped banana peel, chopped longan waste, and chopped rambutan waste (330.6, 268.3, 234.6 and 193.2 mLCH{sub 4}/gVS) that corresponded to their calculated biodegradability. In continuous operations of banana peel digestion at feed concentrations based on total solid (TS) 1–2%, mesophilic single stage digester run at 20-day hydraulic retention time (20-day HRT) failed at 2%TS, but successfully recovered at 1.5%TS. Pre-hydrolysis thermophilic reactor (4-d HRT) was placed as pre-treatment to mesophilic reactor (20-d HRT). Higher biogas (with an evolution of H{sub 2}) and energy yields were obtained and greater system stability was achieved over the single stage digestion, particularly at higher solid feedstock. The best performance of two stage digestion was 68.5% VS destruction and energy yield of 2510.9 kJ/kgVS added at a feed concentration of 2%TS.

09 BIOMASS FUELS↗

Generation of reactive oxygen species (ROS) is a key factor for stimulation of macrophage proliferation by ceramide 1-phosphate

We previously demonstrated that ceramide 1-phosphate (C1P) is mitogenic for fibroblasts and macrophages. However, the mechanisms involved in this action were only partially described. Here, we demonstrate that C1P stimulates reactive oxygen species (ROS) formation in primary bone marrow-derived macrophages, and that ROS are required for the mitogenic effect of C1P. ROS production was dependent upon prior activation of NADPH oxidase by C1P, which was determined by measuring phosphorylation of the p40phox subunit and translocation of p47phox from the cytosol to the plasma membrane. In addition, C1P activated cytosolic calcium-dependent phospholipase A{sub 2} and protein kinase C-{alpha}, and NADPH oxidase activation was blocked by selective inhibitors of these enzymes. These inhibitors, and inhibitors of ROS production, blocked the mitogenic effect of C1P. By using BHNB-C1P (a photolabile caged-C1P analog), we demonstrate that all of these C1P actions are caused by intracellular C1P. It can be concluded that the enzyme responsible for C1P-stimulated ROS generation in bone marrow-derived macrophages is NADPH oxidase, and that this enzyme is downstream of PKC-{alpha} and cPLA{sub 2}-{alpha} in this pathway. -- Highlights: Black-Right-Pointing-Pointer Ceramide 1-phosphate (C1P) stimulates reactive oxygen species (ROS) formation. Black-Right-Pointing-Pointer The enzyme responsible for ROS generation by C1P in macrophages is NADPH oxidase. Black-Right-Pointing-Pointer NADPH oxidase lies downstream of cPLA{sub 2}-{alpha} and PKC-{alpha} in this pathway. Black-Right-Pointing-Pointer ROS generation is essential for the stimulation of macrophage proliferation by C1P.

60 APPLIED LIFE SCIENCES↗

Vascular endothelial growth factor-D is a key molecule that enhances lymphatic metastasis of soft tissue sarcomas

Studies on lymph node metastasis of soft tissue sarcomas are insufficient because of its rarity. In this study, we examined the expressions of vascular endothelial growth factor (VEGF)-C and VEGF-D in soft tissue sarcomas metastasized to lymph nodes. In addition, the effects of the two molecules on the barrier function of a lymphatic endothelial cell monolayer against sarcoma cells were analyzed. We examined 7 patients who had soft tissue sarcomas with lymph node metastases and who had undergone neither chemotherapy nor radiotherapy before lymphadenectomy. Immunohistochemistry revealed that 2 of 7 sarcomas that metastasized to lymph nodes expressed VEGF-C both in primary and metastatic lesions. On the other hand, VEGF-D expression was detected in 4 of 7 primary and 7 of 7 metastatic lesions, respectively. Interestingly, 3 cases that showed no VEGF-D expression at primary sites expressed VEGF-D in metastatic lesions. Recombinant VEGF-C at 10{sup -8} and VEGF-D at 10{sup -7}and 10{sup -8} g/ml significantly increased the random motility of lymphatic endothelial cells compared with controls. VEGF-D significantly increased the migration of sarcoma cells through lymphatic endothelial monolayers. The fact that VEGF-D induced the migration of fibrosarcomas through the lymphatic endothelial monolayer is the probable reason for the strong relationship between VEGF-D expression and lymph node metastasis in soft tissue sarcomas. The important propensities of this molecule for the increase of lymph node metastases are not only lymphangiogenesis but also down-regulation of the barrier function of lymphatic endothelial monolayers, which facilitates sarcoma cells entering the lymphatic circulation.

60 APPLIED LIFE SCIENCES↗

Differential contribution of key metabolic substrates and cellular oxygen in HIF signalling

Changes in availability and utilisation of O{sub 2} and metabolic substrates are common in ischemia and cancer. We examined effects of substrate deprivation on HIF signalling in PC12 cells exposed to different atmospheric O{sub 2}. Upon 2–4 h moderate hypoxia, HIF-α protein levels were dictated by the availability of glutamine and glucose, essential for deep cell deoxygenation and glycolytic ATP flux. Nuclear accumulation of HIF-1α dramatically decreased upon inhibition of glutaminolysis or glutamine deprivation. Elevation of HIF-2α levels was transcription-independent and associated with the activation of Akt and Erk1/2. Upon 2 h anoxia, HIF-2α levels strongly correlated with cellular ATP, produced exclusively via glycolysis. Without glucose, HIF signalling was suppressed, giving way to other regulators of cell adaptation to energy crisis, e.g. AMPK. Consequently, viability of cells deprived of O{sub 2} and glucose decreased upon inhibition of AMPK with dorsomorphin. The capacity of cells to accumulate HIF-2α decreased after 24 h glucose deprivation. This effect, associated with increased AMPKα phosphorylation, was sensitive to dorsomorphin. In chronically hypoxic cells, glutamine played no major role in HIF-2α accumulation, which became mainly glucose-dependent. Overall, the availability of O{sub 2} and metabolic substrates intricately regulates HIF signalling by affecting cell oxygenation, ATP levels and pathways involved in production of HIF-α. - Highlights: • Gln and Glc regulate HIF levels in hypoxic cells by maintaining low O{sub 2} and high ATP. • HIF-α levels under anoxia correlate with cellular ATP and critically depend on Glc. • Gln and Glc modulate activity of Akt, Erk and AMPK, regulating HIF production. • HIF signalling is differentially inhibited by prolonged Glc and Gln deprivation. • Unlike Glc, Gln plays no major role in HIF signalling in chronically hypoxic cells.

60 APPLIED LIFE SCIENCES↗