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At least 595 records · Page 33

Identifying the gluc-1 and gluc-2 mutations in Neurospora crassa by genome resequencing

Genome resequencing is an efficient strategy for associating mutant phenotypes with physical genomic loci. A pilot study of this approach demonstrated that the Neurospora crassa genetic map was critical in narrowing the possible candidate mutations in a strain to a small number in a limited, defined region of the genome. Here in this study, we utilize a resequencing strategy to identify the mutations underlying the gluc-1 and gluc-2 genes in N. crassa.

59 BASIC BIOLOGICAL SCIENCES↗

Grafted nickel-promoter catalysts for dry reforming of methane identified through high-throughput experimentation

High-throughput synthesis of a series of monometallic and bimetallic catalysts (45 bimetallic and 50 monometallic samples) consisting of nickel and one of nine different metal promoters (B, Co, Cu, Fe, Mg, Mn, Sn, V and Zn) supported on one of six different metal oxides alumina, ceria, magnesia, silica and titania) is carried out via organometallic grafting using a robotic platform. The catalysts are evaluated for their activity and selectivity for the dry reforming of methane at a feed ratio of CH 4 :CO 2 of 1 at 650–800 °C in a parallel flow reactor system. The type of oxide support prevails over the type of additive for both catalyst activity and stability. On Al 2 O 3 and MgO, Fe was found to be the best promoter; on SiO 2 , Cu is the best promoter at 700 °C and higher, while on TiO 2 , Mn is found to enhance the conversion at 800 °C. On CeO 2 , all additives except Fe have beneficial effects. Twenty-five catalysts show > 90% methane conversion with ten catalysts showing > 95% conversion at 800 °C with the H 2 :CO ratios ranging from 0.8 to 1.2. Amongst the ten highest performers, NiFe/Al 2 O 3 and NiFe/MgO are more active than Ni/Al 2 O 3 and Ni/MgO, respectively and were stable over a period of 25 h at 800 °C. Characterization on the as-prepared samples reveals highly dispersed phase, while after reduction in H 2 , highly dispersed and reduced nickel particles up to 10 nm are formed. The particles do not increase in size under dry reforming reaction conditions at 800 °C. An increased hydrogen consumption observed during H 2 -TPR of the nickel particles is positively correlated with methane conversion for Al 2 O 3 -based catalysts. The resistance to deactivation by coking and variation in coke structure are investigated by spectroscopic and microscopic methods to identify the relationship between metal promoters, alloy formation, and type of surface carbon deposits. Carbon whiskers were observed on the ten selected spent samples and are preferentially deposited on Ni rather than on the promoters. Carbon nanotube formation and metal particle removal from support were not observed to cause deactivation while amorphous carbon formation was clearly linked to catalyst deactivation, as amorphous carbon could encapsulate nickel, either on the support or at the end of the carbon nanotube. Furthermore, the organometallic grafting technique is an efficient and suitable technique for synthesizing highly dispersed and homogeneous phases which lead to high conversion and high durability for dry reforming of methane.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Utilizing fluorescent life time imaging microscopy technology for identify carriers of BRCA2 mutation

Worldwide, more than one million women are diagnosed with breast cancer every year, making it the most common female malignancy in the developed world. Germline mutations in BRCA1 and BRCA2 genes are estimated to increase the risk for developing breast cancer by up to 87%. From a clinical point of view, identification of BRCA1 and BRCA2 mutation carriers offers an opportunity to early identify or prevent the development of malignancy; therefore the ability to determine which women are more likely to carry BRCA1 or BRCA2 mutations is of great importance. The available diagnostic tests for mutation analysis of BRCA1 and BRCA2 are time- and labor-intensive, expensive, and do not allow the identification of all the functional mutations. We utilized the Fluorescent lifetime (FLT) imaging microscopy method which allows recognizing different cell populations, in order to distinguish between lymphocytes from BRCA1 and BRCA2 mutation carriers and non-carrier women by using easily obtainable lymphocyte cells from peripheral blood. Our results demonstrate that cells originated from BRCA2-mutation carriers have significantly lower FLT values compared with BRCA1 mutation carriers and control cells. This simple, inexpensive and sensitive method may be utilized in the future to detect BRCA2 mutation carriers, particularly those bearing unknown functional mutations. - Highlights: • Fluorescent lifetime imaging microscopy (FLIM) technology can be used for recognizing BRCA mutation carriers. • Cells originated from BRCA2-mutation carriers have lower FLT values compared with BRCA1 mutation carriers and control cells. • The difference in FLT values were observed regardless irradiation. • FLIM method may be more informative in the detection of BRCA2-mutation carriers than BRCA1.

60 APPLIED LIFE SCIENCES↗

Structural study of the effects of mutations in proteins to identify the molecular basis of the loss of local structural fluidity leading to the onset of autoimmune diseases

Protein-Protein Interactions (PPIs) are crucial in most of the biological processes and PPI dysfunctions are known to be associated with the onsets of various diseases. One of such diseases is the auto-immune disease. Auto-immune diseases are one among the less studied group of diseases with very high mortality rates. Thus, we tried to correlate the appearances of mutations with their probable biochemical basis of the molecular mechanisms leading to the onset of the disease phenotypes. We compared the effects of the Single Amino Acid Variants (SAVs) in the wild type and mutated proteins to identify any structural deformities that might lead to altered PPIs leading ultimately to disease onset. For this we used Relative Solvent Accessibility (RSA) as a spatial parameter to compare the structural perturbation in mutated and wild type proteins. We observed that the mutations were capable to increase intra-chain PPIs whereas inter-chain PPIs would remain mostly unaltered. This might lead to more intra-molecular friction causing a deleterious alteration of protein's normal function. A Lyapunov exponent analysis, using the altered RSA values due to polymorphic and disease causing mutations, revealed polymorphic mutations have a positive mean value for the Lyapunov exponent while disease causing mutations have a negative mean value. Thus, local spatial stochasticity has been lost due to disease causing mutations, indicating a loss of structural fluidity. The amino acid conversion plot also showed a clear tendency of altered surface patch residue conversion propensity than polymorphic conversions. So far, this is the first report that compares the effects of different kinds of mutations (disease and non-disease causing polymorphic mutations) in the onset of autoimmune diseases. - Highlights: • Protein-Protein Interaction. • Changes in Relative Solvent Accessibility (RSA). • Amino acid conversion matrix. • Polymorphic mutations. • Disease causing mutations.

60 APPLIED LIFE SCIENCES↗

Reprint of: A chemical screen identifies trifluoperazine as an inhibitor of glioblastoma growth

Highlights: • Screening of GBM cells uncovers 45 active dopaminergic ligands and 8 were validated. • Ligand selectivity suggests dopamine-mediated cell viability regulation in GBM. • The DRD2 antagonist trifluoperazine inhibits cell viability and growth of GBM cells. • DR expression profile of GBM cells alter the susceptibility to trifluoperazine. Glioblastoma (GBM) is regarded as the most common malignant brain tumor but treatment options are limited. Thus, there is an unmet clinical need for compounds and corresponding targets that could inhibit GBM growth. We screened a library of 80 dopaminergic ligands with the aim of identifying compounds capable of inhibiting GBM cell line proliferation and survival. Out of 45 active compounds, 8 were further validated. We found that the dopamine receptor D2 antagonist trifluoperazine 2HCl inhibits growth and proliferation of GBM cells in a dose dependent manner. Trifluoperazine’s inhibition of GBM cells is cell line dependent and correlates with variations in dopamine receptor expression profile. We conclude that components of the dopamine receptor signaling pathways are potential targets for pharmacological interventions of GBM growth.

60 APPLIED LIFE SCIENCES↗

The malaria parasite RhopH protein complex interacts with erythrocyte calmyrin identified from a comprehensive erythrocyte protein library

Highlights: • WGCFS generated protein libraries are useful in identification of host receptors. • Mouse calmyrin is PyRhopH complex RBC receptor in rodent malaria model, P. yoelii. • WGCFS and AlphaScreen technology are invaluable to post-genome malaria research. Malaria merozoite apical organelles; microneme and rhoptry secreted proteins play functional roles during and following invasion of host erythrocytes. Among numerous proteins, the rhoptries discharge high molecular weight proteins known as RhopH complex. Recent reports suggest that the RhopH complex is essential for growth and survival of the malaria parasite within erythrocytes. However, an in-depth understanding of the host-parasite molecular interactions is indispensable. Here we utilized a comprehensive mouse erythrocyte protein library consisting of 443 proteins produced by a wheat germ cell-free system, combined with AlphaScreen technology to identify mouse erythrocyte calmyrin as an interacting molecule of the rodent malaria parasite Plasmodium yoelii RhopH complex (PyRhopH). The PyRhopH interaction was dependent on the calmyrin N-terminus and divalent cation capacity. The finding unveils a recommendable and invaluable usefulness of our comprehensive mouse erythrocyte protein library together with the AlphaScreen technology in investigating a wide-range of host-parasite molecular interactions.

60 APPLIED LIFE SCIENCES↗

Identifying the SUMO1 modification of FAM122A leading to the degradation of PP2A-Cα by ubiquitin-proteasome system

Highlights: • FAM122A can be modified by SUMO1 and the sumoylation is desumoylated by SENP1. • The SUMO1 modification site of FAM122A is lysine 89. • The sumoylated FAM122A degrades PP2A-Cα protein by ubiquitin-proteasome system. • The sumoylated FAM122A reduces the activity of PP2A and suppresses cell proliferations. FAM122A is a highly conserved protein in mammals. Here, we identify that FAM122A can be sumoylated at lysine 89, which can be de-conjugated by SENP1. Furthermore, the sumoylation of FAM122A reduces the PP2A-Cα protein level together with the reduced phosphatase activity of PP2A, which suppresses cell proliferation. Collectively, our results suggest that the sumoylation of FAM122A may have a significant role in cellular function.

60 APPLIED LIFE SCIENCES↗

A natural compound, aristoyagonine, is identified as a potent bromodomain inhibitor by mid-throughput screening

Bromodomain-containing protein 4 (Brd4) is known to play a key role in tumorigenesis. It binds acetylated histones to regulate the expression of numerous genes. Because of the importance of brd4 in tumorigenesis, much research has been undertaken to develop brd4 inhibitors with therapeutic potential. As a result, various scaffolds for bromodomain inhibitors have been identified. To discover new scaffolds, we performed mid-throughput screening using two different enzyme assays, alpha-screen and ELISA. We found a novel bromodomain inhibitor with a unique scaffold, aristoyagonine. This natural compound showed inhibitory activity in vitro and tumor growth inhibition in a Ty82-xenograft mouse model. In addition, we tested Brd4 inhibitors in gastric cancer cell lines, and found that aristoyagonine exerted cytotoxicity not only in I-BET-762-sensitive cancer cells, but also in I-BET-762-resistant cancer cells. This is the first paper to describe a natural compound as a Brd4 bromodomain inhibitor.

60 APPLIED LIFE SCIENCES↗

A parametric approach to identify synergistic domains of process intensification for reactive separation

Process intensification aims to combine multiple tasks within multi-functional units to drastically improve economic, energy or sustainability metrics of a chemical process. Limited work exists to systematically identify the synergistic domains where intensification outperforms its nonintensified counterpart. In this work, we computationally derive the synergistic domains of a reactive separation system. Specifically, we first postulate general models for both intensified and nonintensified systems. We use these models to generate data to train a ReLU-type artifical neural network (ANN). Further, the trained ReLU-NN model is formulated as a multi-parametric mixed-integer linear program (mp-MILP), and the critical regions of this mp-MILP define the synergistic feasible domains of intensification. We have derived these synergistic domains of vapor–liquid equilibrium (VLE)-based reactive separation for several industrial applications. These synergistic domains enable quick screening of properties that favor intensification.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Comprehensive proteomics analysis of stressed human islets identifies GDF15 as a target for type 1 diabetes intervention

Type 1 diabetes results from the progressive loss of beta cells, a process propagated by pro-inflammatory cytokine signaling that disrupts the balance between the expression of pro- and anti-apoptotic proteins. To identify such proteins, we performed comprehensive proteomics of human pancreatic islets treated with interleukin-1ß and interferon-?, leading to the identification of 11,325 proteins, of which 387 were significantly regulated by treatment. We then tested the function of growth/differentiation factor 15 (GDF15), which was repressed by the cytokine treatment. We found that GDF15 translation was blocked during inflammation, while the addition of exogenous GDF15 inhibited interleukin-1ß/interferon-?-induced apoptosis of human islets and a human beta-cell line. Furthermore, administration of GDF15 significantly decreased insulitis in NOD mice. Our approach provides a unique resource for the identification of the human islet proteins regulated by cytokines and was effective in discovering a potential target for type 1 diabetes therapy.

Nakayasu, Ernesto S.↗

Analyzing acoustic emission data to identify cracking modes in cement paste using an artificial neural network

This research is focused on the identification of cracking mechanisms for cement paste using acoustic emission data, recorded from compression and notched four-point bending tests. A procedure is developed for analyzing the data by employing an agglomerative hierarchical clustering method, an artificial neural network, and a ray-tracing source location algorithm. An agglomerative hierarchical clustering method is utilized to cluster the AE data from a compression test using frequency-dependent features. A neural network is trained using the compression test data and applied to the AE data emitted during the four-point bending test. The clustered data from the four-point bending test is localized using a ray-tracing algorithm. Based on the occurrence and locations of the clustered events and signal feature analyses, potential cracking mechanisms are identified and assigned.

36 MATERIALS SCIENCE↗

Implementation of data citations and persistent identifiers at the ORNL DAAC

A requirement of data archives is that data holdings can be easily discovered, accessed, and used. One approach to improving data discovery and access is through data citations coupled with Digital Object Identifiers (DOI). The Oak Ridge National Laboratory Distributed Active Archive Center (ORNL DAAC) since 1998 has issued data citations that have been accepted and used in peer-reviewed journals. Citation elements established by the ORNL DAAC are similar to those used for journal articles (authors, year, product title, and information to locate), and beginning in 2007 included a DOI that is persistent, actionable, specific, and complete. The approach used at the ORNL DAAC also allows for referring to subsets of the data, by including within the citation the temporal and spatial extent, and parameters used. Data citations allow readers to find data and reproduce the results of the research article, and also use those data to test new hypotheses, design new sample collections, or construct or evaluate models. The ORNL DAAC uses a manual method to compile data citations and has developed a database that links research articles and their use of specific ORNL DAAC data products. Automation of the data citation compilation process, as is the case for articles, will enable data citations to become a more common practice. In addition to enhancing discovery and access of the data used in a research article, the citation gives credit to data generators, data centers and their funders, and, through citation indices, determine the scientific impact of a data set.

96 KNOWLEDGE MANAGEMENT AND PRESERVATION↗

A temperature threshold to identify the driving climate forces of the respiratory process in terrestrial ecosystems

Terrestrial ecosystem respiration (R e ) is the second largest carbon flux between the biosphere and atmosphere. Therefore, climate-driven changes of Re greatly impact on future atmospheric CO 2 concentration. The aim of this study was to derive an air temperature threshold to identify the driving climate forces of the respiratory process in terrestrial ecosystems within different temperature zones. A global dataset of 647 site-years of ecosystem flux data and related variables were collected at 152 sites. The quantile regression was applied to evaluate relationships between the maximum realizable Re rates and mean annual air temperature (MAT) as well as other micrometeorological factors (i.e., atmospheric CO 2 concentration, atmospheric water content, soil heat flux, sensible heat flux, latent heat flux, precipitation, relative humidity, and soil water content). Our analysis revealed an ecosystem threshold of MAT of 11 ± 2.3 °C. In ecosystems with MATs lower than the threshold, the maximum R e rates were primarily dependent on temperature and respiration was mainly a temperature-driven process. In ecosystems with MATs higher than the threshold, besides MAT, other factors, such as water availability and surface heat flux, became significant driving forces of the maximum Re rates and respiration was a multi-factor-driven process. Temperature played the key role in generation of the maximum R e rates in the terrestrial ecosystems, while other driving forces reduce the maximum R e rates and the temperature sensitivity of the respiratory process. According to a regression tree analysis, MAT was also the most influencing factor of mean R e rates among the climate forces. The information from this study should be useful to predict the respiratory process in terrestrial ecosystems with different temperatures under the climate change.

54 ENVIRONMENTAL SCIENCES↗

A practical approach for identifying various polarization behaviors of redox-stable electrodes in symmetrical solid oxide fuel cells

In symmetrical solid oxide fuel cells, comprehensively understanding the elementary reaction processes and the polarization behaviors of redox-stable electrode materials is critical for further optimization of the electrode performance. In this work, a systematical and practical approach, based on electrochemical impedance spectroscopy technology, is applied to identify the rate-limiting elementary reactions of the redox-stable electrodes. The feasibility of this proposed method is demonstrated in symmetrical solid oxide fuel cells with Sr 2 Fe 1.5 Mo 0.5 O 6-σ -Ce 0.9 Gd 0.1 O 1.95 as electrodes. Based on the characteristic frequency ranges and the experimental results tested under various fuel gas components, operating temperatures, and discharge current densities, the rate-limiting steps of the cathode are associated with the formation of adsorbed oxygen ions and the combination of oxygen ions and oxygen vacancies, while the rate-limiting steps of the anode are ascribed to the hydrogen dissociated adsorption and the steam desorption processes. This experimental and analysis framework can be straightforwardly extended to other electrode materials to unravel their electrochemical performance in detail.

25 ENERGY STORAGE↗

Personal and environmental predictors of polycyclic aromatic hydrocarbon exposure identified through repeated silicone wristband sampling

This study integrates quantitative data on personal exposure to polycyclic aromatic hydrocarbons (PAHs) in 162 silicone wristbands with demographics, behavioral information, and housing characteristics to explore contributions to residential exposure in a superfund-adjacent community over the course of a year. Forty-six residents completed questionnaires and wore silicone wristbands as personal passive samplers for seven consecutive days on up to four separate occasions in alternating months between November 2022 and June 2023. It was hypothesized that individual behaviors and housing characteristics are sources of dependence and correlation between personal PAH exposures. 50 PAHs were detected at least once, 17 of which were alkylated PAHs. Exposure to PAHs of similar molecular weight was often correlated, notably between naphthalenes (2-rings) and higher molecular weight PAHs (3 or more rings). Generalized linear mixed models identified flooring type, participant age, and sampling month as important predictors of increased PAH exposure, and flooring type, and use of wood stoves or heavy machinery as predictors of increased naphthalene exposure relative to higher molecular weight PAHs. Individual chemical models based on concentration data and detection frequencies corroborated these findings across multiple PAHs. We demonstrate that personal exposure is not static and the degree of variability in personal exposure is individual. Hence, identification of influential exposure factors through repeated measures of chemical exposure and characterization of variability in personal exposure as performed in this study, is important in the development of exposure mitigation strategies.

Bonner, Emily↗

Insights from an international stakeholder consultation to identify informational needs related to seafood safety

Food safety assessment and communication have a strong importance in reducing human health risks related to food consumption. The research carried out within the ECsafeSEAFOOD project aims to assess seafood safety issues, mainly related to non-regulated priority environmental contaminants, and to evaluate their impact on public health. In order to make the research results accessible and exploitable, and to respond to actual stakeholders' demands, a consultation with international stakeholders was performed by means of a survey. The focus was on policy and decision makers, food producers and processors, and agencies (i.e. EU and National or Regional agencies related to Food Safety or Public Health) and consumer organisations. The survey considered questions related to: seafood safety assessment and mitigation strategies, availability of data, such as the level of information on different contaminants, and communication among different stakeholder groups. Furthermore, stakeholders were asked to give their opinion on how they believe consumers perceive risks associated with environmental contaminants. The survey was distributed to 531 key stakeholders and 91 responses were received from stakeholders from 30 EU and non-EU countries. The main results show that communication between different groups of stakeholders needs to be improved and that there is a deficit of information and data in the field of seafood safety. This pertains mainly to the transfer of contaminants between the environment and seafood, and to the diversity of environmental contaminants such as plastic additives, algal toxins and hormones. On-line tools were perceived to be the most useful communication channel. - Highlights: • We consulted stakeholders to identify their needs about seafood safety. • An on-line survey was prepared and sent to gather stakeholders' opinions. • Communication among stakeholders needs to be improved. • There is a deficit of information and data in the field of seafood safety. • On-line tools are perceived to be the most useful communication channel.

54 ENVIRONMENTAL SCIENCES↗

Geochemistry and mercury contamination in receiving environments of artisanal mining wastes and identified concerns for food safety

Artisanal small-scale gold mining (ASGM) using mercury (Hg) amalgamation has been occurring on Buru Island, Indonesia since early 2012, and has caused rapid accumulation of high Hg concentrations in river, estuary and marine sediments. In this study, sediment samples were collected from several sites downstream of the Mount Botak ASGM site, as well as in the vicinity of the more recently established site at Gogrea where no sampling had previously been completed. All sediment samples had total Hg (THg) concentrations exceeding Indonesian sediment quality guidelines and were up to 82 times this limit at one estuary site. The geochemistry of sediments in receiving environments indicates the potential for Hg-methylation to form highly bioavailable Hg species. To assess the current contamination threat from consumption of local seafood, samples of fish, molluscs and crustaceans were collected from the Namlea fish market and analysed for THg concentrations. The majority of edible tissue samples had elevated THg concentrations, which raises concerns for food safety. This study shows that river, estuary and marine ecosystems downstream of ASGM operations on Buru Island are exposed to dangerously high Hg concentrations, which are impacting aquatic food chains, and fisheries resources. Considering the high dietary dependence on marine protein in the associated community and across the Mollucas Province, and the short time period since ASGM operations commenced in this region, the results warrant urgent further investigation, risk mitigation, and community education. - Highlights: • Mercury contamination of sediments and seafood due to artisanal gold mining. • Considerable risks to human and ecosystem health are identified. • Results emphasise the urgent need for risk mitigation and community education.

54 ENVIRONMENTAL SCIENCES↗

Identifying geochemical hot moments and their controls on a contaminated river floodplain system using wavelet and entropy approaches

Geochemical hot moments are defined here as short periods of time that are associated with disproportionally high levels of concentrations (biogeochemically-driven or transport-related) relative to longer intervening time periods. We used entropy and wavelet techniques to identify temporal variability in geochemical constituents and their controls along three transects within a contaminated floodplain system near Rifle CO. Results indicated that transport-dominated hot moments drove overall geochemical processing in the contaminated groundwater and seep zones. These hot moments were associated with seasonal hydrologic variability (~4 months) in the contaminated aquifer and with annual hydrologic cycle and residence times in the seep zone. Hot moments associated with a naturally reduced zone within the aquifer were found to be biogeochemically-driven, with a different dominant frequency (~3 months) and no correlation to hydrologic or weather variations, in contrast to what is observed in other regions of the floodplain.

54 ENVIRONMENTAL SCIENCES↗