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At least 631 records · Page 35

ClbM is a versatile, cation-promiscuous MATE transporter found in the colibactin biosynthetic gene cluster

Multidrug transporters play key roles in cellular drug resistance to toxic molecules, yet these transporters are also involved in natural product transport as part of biosynthetic clusters in bacteria and fungi. The genotoxic molecule colibactin is produced by strains of virulent and pathobiont Escherichia coli and Klebsiella pneumoniae. In the biosynthetic cluster is a multidrug and toxic compound extrusion protein (MATE) proposed to transport the prodrug molecule precolibactin across the cytoplasmic membrane, for subsequent cleavage by the peptidase ClbP and cellular export. We recently determined the X-ray structure of ClbM, and showed preliminary data suggesting its specific role in precolibactin transport. Here, we define a functional role of ClbM by examining transport capabilities under various biochemical conditions. Our data indicate ClbM responds to sodium, potassium, and rubidium ion gradients, while also having substantial transport activity in the absence of alkali cations. - Highlights: • ClbM is a cation promiscuous MATE multidrug transporter. • The role of key residues were identified in both the cation and proton binding. • The biologically relevant substrate for ClbM is the natural product precolibactin.

60 APPLIED LIFE SCIENCES↗

Adenovirus infection induces HuR relocalization to facilitate virus replication

HuR is an RNA-binding protein of the embryonic lethal abnormal vision (ELAV) family, which binds to the AU-rich element (ARE) in the 3′-untranslated region (UTR) of certain mRNAs and is involved in the nucleo-cytoplasmic export and stabilization of ARE-mRNAs. The cytoplasmic relocalization of ARE-mRNAs with several proteins such as HuR and pp32 increases in cells transformed by the adenovirus oncogene product E4orf6. Additionally, these ARE-mRNAs were stabilized and acquired the potential to transform cells. Although, the relocalization of HuR and the stabilization of ARE-mRNAs are crucial for cell transformation, evidence regarding the relationship of HuR and ARE-mRNAs with adenovirus replication is lacking. In this report, we demonstrate that adenovirus infection induces the relocation of HuR to the cytoplasm of host cells. Analysis using the luciferase-ARE fusion gene and the tetracycline (tet)-off system revealed that the process of stabilizing ARE-mRNAs is activated in adenovirus-infected cells. Heat shock treatment or knockdown-mediated depletion of HuR reduced adenovirus production. Furthermore, expression of ARE-including viral IVa2 mRNA, decreased in HuR-depleted infected cells. These results indicate that HuR plays an important role in adenovirus replication, at least in part, by up-regulating IVa2 mRNA expression and that ARE-mRNA stabilization is required for both transformation and virus replication.

60 APPLIED LIFE SCIENCES↗

Seed-specific overexpression of AtFAX1 increases seed oil content in Arabidopsis

Highlights: • Seed-specific overexpression of AtFAX1 increased seed oil and protein content in Arabidopsis. • Fatty acid and triacylglycerol biosynthesis activity were enhanced in AtFAX1 transgenic plants. • Genes involved in seed sugar importation were induced in AtFAX1 transgenic plants. • Seed-specific overexpression of AtFAX1 increased seed mass and seed size. Biosynthesis of plant seed oil is accomplished through the coordinate action of multiple enzymes in multiple subcellular compartments. Fatty acid (FA) has to be transported from plastid to endoplasmic reticulum (ER) for TAG synthesis. However, the role of plastid FA transportation during seed oil accumulation has not been evaluated. AtFAX1 (Arabidopsis fatty acid export1) mediated the FA export from plastid. In this study, we overexpressed AtFAX1 under the control of a seed specific promoter in Arabidopsis. The resultant overexpression lines (OEs) produced seeds which contained 21–33% more oil and 24–30% more protein per seed than those of the wild type (WT). The increased oil content was probably because of the enhanced FA and TAG synthetic activity. The seed size and weight were both increased accordingly. In addition, the seed number per silique and silique number per plant had no changes in transgenic plants. Taken together, our results demonstrated that seed specific overexpression of AtFAX1 could promote oil accumulation in Arabidopsis seeds and manipulating FA transportation is a feasible strategy for increasing the seed oil content.

60 APPLIED LIFE SCIENCES↗

Designing an effective drug combination for ER stress loading in cancer therapy using a real-time monitoring system

Excess stress caused by accumulation of misfolded proteins inside the endoplasmic reticulum (ER) lumen can cause cells to undergo apoptosis. Misfolded proteins exported from ER to cytoplasm are ubiquitinated and mostly degraded by the proteasome, but can also be destroyed by autophagy mediated by the docking proteins p62 and NBR1. When misfolded proteins accumulate beyond the capacity of these clearance systems, they are transported to the microtubule organization center to form aggresomes, which are also degraded by autophagy. Together, these phenomena suggest the existence of a coordinated intracellular network for coping with the accumulation of misfolded proteins. Thus, rational inhibition of this network system might enhance killing of cancer cells subjected to pronounced ER stress loading. Based on this rationale, we sought to establish a quantitative assay for monitoring ER stress loading. MDA-MB231 cells stably transfected with the ERAI-Venus vector exhibited a strong XBP1 splicing signal in response to ER stress. Using the IncuCyte cell imaging system, we monitored the fluorescence intensity of XBP1-Venus, normalized against cell density, as an ER stress indicator. This parameter correlated closely with other reporters of unfolded protein responses. Assessment of the XBP1-Venus signal during exposure to various drug combinations revealed that simultaneous inhibition of the proteasome, autophagy, and aggresome formation led to more effective ER stress loading and higher cytotoxicity than inhibition of only two components. Our data suggest that this monitoring system is a useful tool for designing effective drug combinations for ER stress loading in cancer therapy.

60 APPLIED LIFE SCIENCES↗

Role of nucleocytoplasmic transport in interphase microtubule organization in fission yeast

The proper organization of microtubules is essential for many cellular functions. Microtubule organization and reorganization are highly regulated during the cell cycle, but the underlying mechanisms remain elusive. Here we characterized unusual interphase microtubule organization in fission yeast nuclear export mutant crm1-124. The mutant cells have an intranuclear microtubule bundle during interphase that pushes the nuclear envelope to assume a protruding morphology. We showed that the formation of this protruding microtubule bundle requires the nuclear accumulation of two microtubule-associated proteins (MAPs), Alp14/TOG and Mal3/EB1. Interestingly, the forced accumulation of Alp14 in the nucleus of wild type cells is sufficient to form the intranuclear microtubule bundle. Furthermore, the frequency of the intranuclear microtubule formation by Alp14 accumulated in the nucleus is prominently increased by a reduction in the nucleation activity of interphase cytoplasmic microtubules. We propose that properly regulated nucleocytoplasmic transport and maintained activity of cytoplasmic microtubule nucleation during interphase are important for the proper organization of interphase cytoplasmic microtubules.

60 APPLIED LIFE SCIENCES↗

Nature’s conductors: what can microbial multi-heme cytochromes teach us about electron transport and biological energy conversion?

Microorganisms can acquire energy from the environment by extending their electron transport chains to external solid electron donors or acceptors. This process, known as extracellular electron transfer (EET), is now being heavily pursued for wiring microbes to electrodes in bioelectrochemical renewable energy technologies. Recent studies highlight the crucial role of multi-heme cytochromes in facilitating biotic–abiotic EET both for cellular electron export and uptake. Here we explore progress in understanding the range and function of these biological electron conduits in the context of fuel-to-electricity and electricity-to-bioproduct conversion. We also highlight emerging topics, including the role of multi-heme cytochromes in inter-species electron transfer and in inspiring the design and synthesis of a new generation of protein-based bioelectronic components.

59 BASIC BIOLOGICAL SCIENCES↗

A novel system for ammonia-based sustainable energy and agriculture: Concept and design optimization

Here this work presents a new vision for sustainable energy supply and agriculture: an ammonia-based sustainable energy agriculture (ABSEA) system. This intensified system uses renewables to produce ammonia both as fertilizer and as fuel for tractors and grain drying, to meet local electrical power demand and to provide predictable, consistent power export to the grid. The conceptual design of this system exploits synergies between ammonia production and energy supply to allow for economic viability, specifically time-varying chemical production as well as the potential of ammonia and hydrogen as energy storage media. A framework for combined optimization of the ABSEA design and its inherently time-varying operating schedule is proposed with a view on further improving economic competitiveness. A case study which considers the use of wind turbines to meet ammonia fertilizer and fuel demands of 40,300 kg/year and an annual hourly average power demand of 985 kW demonstrates the promise of the ABSEA system and the associated design-scheduling optimization framework. The annual net present cost is approximately $\$$75,000, which corresponds to an emissions reduction cost of $\$$17.60/tonCO2. The ABSEA optimal design uses both hydrogen and ammonia, but not a battery, for energy storage, illustrating their efficacy in this synergistic application.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

The transport of monomers during lignification in plants: anything goes but how?

Lignin is a highly abundant polymer in plant cell walls that is essential for land plants’ ability to stand upright and transport water. Inside plant cells, lignin monomers, called monolignols, are made from phenylalanine via a multistep pathway. In the cell wall, monomers move freely, until they encounter stationary oxidative enzymes that determine where the lignin polymer forms. However, it remains unclear how lignin monomers are trafficked from inside the cell to the cell wall. Although multiple lines of circumstantial evidence implicate transporters, additional possible mechanisms include the diffusion of monomers across lipid bilayers and the release of monolignol glucosides stored in vacuoles. There are therefore potentially diverse and overlapping mechanisms of monolignol export.

59 BASIC BIOLOGICAL SCIENCES↗

A new database website for nuclear level densities

We introduce a new open-access, web-based database (http://nld.ascsn.net), Current Archive of Nuclear Density of Levels (CANDL), that hosts experimental nuclear level density (NLD) datasets from a variety of techniques and energy ranges. Built using the Dash framework in Python, the database is designed to be interactive and user-friendly, allowing researchers to search, visualize, fit, and export NLD data with minimal effort. This resource includes data extracted from evaporation spectra, Oslo method variants, and other experimental techniques that cover excitation energies beyond the neutron resonance region. The database supports on-the-fly fitting with two widely-used phenomenological models—the Constant Temperature (CT) model and the Back-Shifted Fermi Gas (BSFG) model—selected for their simplicity and computational efficiency. Future versions aim to include additional datasets and model types, as well as easy-to-use interfaces to data science techniques. Here, this platform offers a vital tool for the nuclear physics, astrophysics, medicine, and reactor design communities.

73 NUCLEAR PHYSICS AND RADIATION PHYSICS↗

Dataset of timberland variables used to assess forest conditions in two Southeastern United States' fuelsheds

The data presented in this article are related to the research article entitled “How is wood-based pellet production affecting forest conditions in the southeastern United States?” (Dale et al., 2017). This article describes how United States Forest Service (USFS) Forest Inventory and Analysis (FIA) data from multiple state inventories were aggregated and used to extract ten annual timberland variables for trend analysis in two case study bioenergy fuelshed areas. This dataset is made publically available to enable critical or extended analyses of changes in forest conditions, either for the fuelshed areas supplying the ports of Savannah, Georgia and Chesapeake, Virginia, or for other southeastern US forested areas contributing biomass to the export wood pellet industry.

54 ENVIRONMENTAL SCIENCES↗

Control algorithms for dynamic windows for residential buildings

This study analyzes the influence of control algorithms for dynamic windows on energy consumption, number of hours of retracted shades during daylight and shade operations. Five different control algorithms - heating/cooling, simple rules, perfect citizen, heat flow and predictive weather were developed and compared. The performance of a typical residential building was modeled with EnergyPlus. The program Widow was used to generate a Bi-Directional Distribution Function (BSDF) for two window configurations. The BSDF was exported to EnergyPlus using the IDF file format. The EMS feature in EnergyPlus was used to develop custom control algorithms. The calculations were made for four locations with diverse climate. The results showed that: (a) use of automated shading with proposed control algorithms can reduce the site energy in the range of 11.6-13.0%; in regard to source (primary) energy in the range of 20.1-21.6%, (b) the differences between algorithms in regard to energy savings are not high, (c) the differences between algorithms in regard to number of hours of retracted shades are visible, (e) the control algorithms have a strong influence on shade operation and oscillation of shade can occur, (d) additional energy consumption caused by motor, sensors and a small microprocessor in the analyzed case is very small.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Future energy system challenges for Africa: Insights from Integrated Assessment Models

Although Africa’s share in the global energy system is only small today, the ongoing population growth and economic development imply that this can change significantly. In this paper, we discuss long-term energy developments in Africa using the results of the LIMITS model inter-comparison study. The analysis focusses on the position of Africa in the wider global energy system and climate mitigation. The results show a considerable spread in model outcomes. Without specific climate policy, Africa’s share in global CO 2 emissions is projected to increase from around 1-4% today to 3-23% by 2100. In all models, emissions only start to become really significant on a global scale after 2050. Furthermore, by 2030 still around 50% of total household energy use is supplied through traditional bio-energy, in contrast to existing ambitions from international organisations to provide access to modern energy for all. After 2050, the energy mix is projected to converge towards a global average energy mix with high shares of fossil fuels and electricity use. Finally, although the continent is now a large net exporter of oil and gas, towards 2050 it most likely needs most of its resources to meet its rapidly growing domestic demand. With respect to climate policy, the rapid expansion of the industrial and the power sector also create large mitigation potential and thereby the possibility to align the investment peak in the energy system with climate policy and potential revenues from international carbon trading.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗

Lessons from wind policy in Portugal

Wind capacity and generation grew rapidly in several European countries, such as Portugal. Wind power adoption in Portugal began in the early 2000s, incentivized by a continuous feed-in tariff policy mechanism, coupled with public tenders for connection licenses in 2001, 2002, and 2005. These policies led to an enormous success in terms of having a large share of renewables providing electricity services: wind alone accounts today for ~23.5% of electricity demand in Portugal. We explain the reasons wind power became a key part of Portugal's strategy to comply with European Commission climate and energy goals, and provide a detailed review of the wind feed-in tariff mechanism. We describe the actors involved in wind power production growth. We estimate the environmental and energy dependency gains achieved through wind power generation, and highlight the correlation between wind electricity generation and electricity exports. Finally, we compare the Portuguese wind policies with others countries' policy designs and discuss the relevance of a feed-in tariff reform for subsequent wind power additions.

17 WIND ENERGY↗

Species- and element-specific patterns of metal flux from contaminated wetlands versus metals shed with exuviae in emerging dragonflies

Dragonfly adults and their aquatic immature stages are important parts of food webs and provide a link between aquatic and terrestrial components. During emergence, contaminants can be exported into terrestrial food webs as immature adults fly away or be shed with their exuviae and remain in the wetland. Our previous work established metals accumulating in dragonfly nymphs throughout a contaminated constructed wetland designed to regulate pH and sequester trace metals from an industrial effluent line. In this paper we evaluated the concentration and mass of metals leaving the wetland in flying emergents versus remaining in the wetland with the shed exuviae in 10 species of dragonflies belonging to 8 genera. Nine elements (Cu, Zn, Cd, Mn, V, Mg, Fe, Al, Pb) were evaluated that include essential and nonessential elements as well as trace and major metals. Metal concentrations in the emergent body and exuviae can differ by orders of magnitude. Aluminum, Fe, Mn, and Pb were largely shed in the exuviae. Vanadium and Cd were more variable among species but also tended to be shed with the exuviae. In contrast, Cu, Zn, and Mg showed a higher tendency to leave the wetland with an emerging dragonfly. Metals shed in dragonfly exuviae can moderate the transport of metals from contaminated wetlands. Taxonomic- and metal-specific variability in daily metal flux from the wetland depended upon concentration accumulated, individual body mass, and number of individuals emerging, with each factor's relative importance often differing among species. This illustrates the importance of evaluating the mass of metals in an individual and not only concentrations. Furthermore, differences in numbers of each species emerging will magnify differences in individual metal flux when calculating community metal flux. A better understanding of the variability of metal accumulation in nymphs/larvae and metal shedding during metamorphosis among both metals and species is needed.

54 ENVIRONMENTAL SCIENCES↗

Evaluation of the effect of divalent metal transporter 1 gene polymorphism on blood iron, lead and cadmium levels

Divalent metal transporter 1 (DMT1), a member of the proton-coupled metal ion transporter family, mediates transport of ferrous iron from the lumen of the intestine into the enterocyte and export of iron from endocytic vesicles. It has an affinity not only for iron but also for other divalent cations including manganese, cobalt, nickel, cadmium, lead, copper, and zinc. DMT1 is encoded by the SLC11a2 gene that is located on chromosome 12q13 in humans and express four major mammalian isoforms (1A/+IRE, 1A/-IRE, 2/+IRE and 2/-IRE). Mutations or polymorphisms of DMT1 gene may have an impact on human health by disturbing metal trafficking. To study the possible association of DMT1 gene with the blood levels of some divalent cations such as iron, lead and cadmium, a single nucleotide polymorphism (SNP) (IVS4+44C/A) in DMT1 gene was investigated in 486 unrelated and healthy individuals in a Turkish population by method of polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP). The genotype frequencies were found as 49.8% homozygote typical (CC), 38.3% heterozygote (CA) and 11.9% homozygote atypical (AA). Metal levels were analyzed by dual atomic absorption spectrometer system and the average levels of iron, lead and cadmium in the blood samples were 446.01±81.87 ppm, 35.59±17.72 ppb and 1.25±0.87 ppb, respectively. Individuals with the CC genotype had higher blood iron, lead and cadmium levels than those with AA and CA genotypes. Highly statistically significant associations were detected between IVS4+44 C/A polymorphism in the DMT1 gene and iron and lead levels (p=0.001 and p=0.036, respectively), but no association was found with cadmium level (p=0.344). This study suggested that DMT1 IVS4+44 C/A polymorphism is associated with inter-individual variations in blood iron, lead and cadmium levels. - Highlights: • DMT1 IVS4+44 C/A polymorphism is associated with inter-individual variations in blood iron, cadmium and lead levels. • Individuals with the DMT1 IVS4+44 CC genotype had higher blood iron, lead and cadmium levels. • Individuals with DMT1 IVS4+44 CC genotype may be more susceptible for metal toxicity.

54 ENVIRONMENTAL SCIENCES↗

Bioaccumulation of persistent and emerging pollutants in wild sea urchin Paracentrotus lividus

Highlights: • Bioaccumulation of 99 organic contaminants were assessed in wild sea urchin. • 40 emerging and persistent organic contaminants were detected, generally at low levels. • Differences were found between sampling sites, Carreço being the least impacted area. Marine pollution has been increasing as a consequence of anthropogenic activities. The preservation of marine ecosystems, as well as the safety of harvested seafood, are nowadays a global concern. Here, we report for the first time the contamination levels of a large set of 99 emerging and persistent organic contaminants (butyltins (BTs), polycyclic aromatic hydrocarbons (PAHs), pesticides including pyrethroids, pharmaceuticals and personal care products (PCPs) and flame retardants) in roe/gonads of sea urchin Paracentrotus lividus. Sea urchins are a highly prized worldwide delicacy, and the harvesting of this seafood has increased over the last decades, particularly in South West Atlantic coast, where this organism is harvested mainly for exportation. Sampling was performed in three harvesting sites of the NW Portuguese coast subjected to distinct anthropogenic pressures: Carreço, Praia Norte and Vila Chã, with sea urchins being collected in the north and south areas of each site. Butyltins and pharmaceuticals were not found at measurable levels. Several PAHs, four pyrethroids insecticides, four PCPs and eleven flame retardants were found in roe/gonads of sea urchins, though in general at low levels. Differences among harvesting sites and between areas within each site were found, the lowest levels of contaminants being registered in Carreço. The accumulation of contaminants in sea urchins’ roe/gonads seemed to reflect the low anthropogenic pressure felt in the sampling sites. Nevertheless, taking into account the low accumulated levels of chemicals, results indicate that sea urchins collected in South West Atlantic coast are safe for human consumption.

54 ENVIRONMENTAL SCIENCES↗

Organic sulfur fluxes and geomorphic control of sulfur isotope ratios in rivers

Pyrite oxidation plays a critical role in the relationship between weathering and climate, and its impact on the global carbon cycle has previously been constrained through inversion models utilizing observations of river sulfate ($SO^{2–}_{4}$) and its 34 S/ 32 S isotope ratio (δ 34 S SO4 ). However, measurements from some rivers have suggested that SSO4 can be substantially impacted by processes such as microbial sulfate reduction and/or sulfur assimilation and cycling, rather than simply reflecting a weighted mixture of lithologic sulfur sources. To study the prevalence and controls on $SO^{2–}_{4}$ transformations, in this study we measured dissolved major element concentrations and δ 34 S SO4 in river water samples from throughout western Iceland. Our analyses focused on samples from a small catchment hosting the Efri Haukadalsá river, a system with relatively uniform and isotopically constrained basaltic bedrock. We also measured sediment δ 34 S and sulfur speciation using sulfur K-edge X-ray absorption spectroscopy on sediment and vegetation samples from this catchment. Values of dissolved δ 34 S SO4 in the Efri Haukadalsá ranged from 2.5‰ to 23.7‰ and had a linear relationship with Cl – /$SO^{2–}_{4}$ ratios, indicating that $SO^{2–}_{4}$ predominantly derived from basalt weathering and meteoric precipitation. The lower δ 34 S SO4 values were found in fluvial valleys with V-shaped cross sections, while higher values of δ 34 S SO4 occurred in U-shaped, glacially eroded valleys with thick alluvial fills blanketing the valley floor. Spectroscopic observations identified organic sulfur phases in suspended river sediment, floodplain deposits, and vegetation. Mass balance calculations quantified the organic sulfur flux as less than 10% of $SO^{2–}_{4}$ export, and sediment δ 34 S values were comparable to river δ 34 S SO4 . We interpreted these isotopic and chemical patterns as reflecting differences in the availability of unweathered bedrock across the Efri Haukadalsá catchment, with V-shaped valleys having greater access to fresh sulfide-bearing minerals than alluviated U-shaped valleys; this interpretation is in contrast to one in which the elevated δ 34 S SO4 values reflect fractionation during sulfur transformations along alluvial reaches. These results validated the application of river inversion models for constraining weathering fluxes and affirmed that pyrite oxidation globally, even in the presence of river sulfur cycling, modulates the abundance of atmospheric carbon dioxide.

54 ENVIRONMENTAL SCIENCES↗