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At least 343 records · Page 19

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DOE OSTI2016

PKC-alpha modulation by miR-483-3p in platinum-resistant ovarian carcinoma cells

The occurrence of drug resistance limits the efficacy of platinum compounds in the cure of ovarian carcinoma. Since microRNAs (miRNAs) may contribute to this phenomenon by regulating different aspects of tumor cell response, the aim of this study was to exploit the analysis of expression of miRNAs in platinum sensitive/resistant cells in an attempt to identify potential regulators of drug response. MiR-483-3p, which may participate in apoptosis and cell proliferation regulation, was found up-regulated in 4 platinum resistant variants, particularly in the IGROV-1/Pt1 subline, versus parental cells. Transfection of a synthetic precursor of miR-483-3p in IGROV-1 parental cells elicited a marked up-regulation of the miRNA levels. Growth-inhibition and colony-forming assays indicated that miR-483-3p over-expression reduced cell growth and conferred mild levels of cisplatin resistance in IGROV-1 cells, by interference with their proliferative potential. Predicted targets of miR-483-3p included PRKCA (encoding PKC-alpha), previously reported to be associated to platinum-resistance in ovarian carcinoma. We found that miR-483-3p directly targeted PRKCA in IGROV-1 cells. In keeping with this finding, cisplatin sensitivity of IGROV-1 cells decreased upon molecular/pharmacological inhibition of PKC-alpha. Overall, our results suggest that overexpression of miR-483-3p by ovarian carcinoma platinum-resistant cells may interfere with their proliferation, thus protecting them from DNA damage induced by platinum compounds and ultimately representing a drug-resistance mechanism. The impairment of cell growth may account for low levels of drug resistance that could be relevant in the clinical setting. - Highlights: • miR-483-3p is up-regulated in ovarian carcinoma cells resistant to platinum drugs. • Ectopic expression of miR-483-3p in IGROV-1 confers mild levels of Pt-resistance. • Overexpression of miR-483-3p down-regulates PRKCA levels in ovarian carcinoma cells. • miR 483–3p directly targets PRKCA 3′UTR in IGROV-1 cells. • The contribution of miR-483-3p to Pt-resistance is related to cell proliferation.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2016

Inhibition of autophagy by andrographolide resensitizes cisplatin-resistant non-small cell lung carcinoma cells via activation of the Akt/mTOR pathway

Resistance to cisplatin is a major obstacle for the success of non-small cell lung cancer therapy. The mechanisms underlying cisplatin resistance are not fully understood. In this study, we found that the increase of basal auotophagy accompanied the development of cisplatin resistance. Meanwhile the blockade of the Akt/mTOR pathway occurred in the process. Inhibition of this pathway was induced by cisplatin treatment in the resistant non-small cell lung carcinoma cells. Andrographolide, a natural diterpenoid, promoted the activation of the Akt/mTOR signaling by downregulating PTEN and suppressed autophagy, which subsequently resensitized the resistant cells to cisplatin-mediated apoptosis. Cisplatin treatment in combination with andrographolide significantly prevented the growth of the resistant cells in vivo. These results highlight the involvement of autophagy in cisplatin-resistance development and suggest that inhibition of autophagy via tuning the Akt/mTOR signaling could be a promising strategy in the therapy for cisplatin-resistant non-small cell lung cancer. - Highlights: • The increase of basal auotophagy accompanied the development of cisplatin resistance in NSCLC cells. • Cisplatin induced the blockade of the Akt/mTOR pathway. • Andrographolide promoted the activation of the Akt/mTOR signaling. • Andrographolide downregulated PTEN expression. • Cisplatin treatment in combination with andrographolide resensitized the resistant cells to cisplatin.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2016

Nuclear respiratory factor-1 and bioenergetics in tamoxifen-resistant breast cancer cells

Acquired tamoxifen (TAM) resistance is a significant clinical problem in treating patients with estrogen receptor α (ERα)+ breast cancer. We reported that ERα increases nuclear respiratory factor-1 (NRF-1), which regulates nuclear-encoded mitochondrial gene transcription, in MCF-7 breast cancer cells and NRF-1 knockdown stimulates apoptosis. Whether NRF-1 and target gene expression is altered in endocrine resistant breast cancer cells is unknown. We measured NRF-1and metabolic features in a cell model of progressive TAM-resistance. NRF-1 and its target mitochondrial transcription factor A (TFAM) were higher in TAM-resistant LCC2 and LCC9 cells than TAM-sensitive MCF-7 cells. Using extracellular flux assays we observed that LCC1, LCC2, and LCC9 cells showed similar oxygen consumption rate (OCR), but lower mitochondrial reserve capacity which was correlated with lower Succinate Dehydrogenase Complex, Subunit B in LCC1 and LCC2 cells. Complex III activity was lower in LCC9 than MCF-7 cells. LCC1, LCC2, and LCC9 cells had higher basal extracellular acidification (ECAR), indicating higher aerobic glycolysis, relative to MCF-7 cells. Mitochondrial bioenergetic responses to estradiol and 4-hydroxytamoxifen were reduced in the endocrine-resistant cells compared to MCF-7 cells. These results suggest the acquisition of altered metabolic phenotypes in response to long term antiestrogen treatment may increase vulnerability to metabolic stress. - Highlights: • NRF-1 and TFAM expression are higher in endocrine-resistant breast cancer cells. • Oxygen consumption rate is similar in endocrine-sensitive and resistant cells. • Mitochondrial reserve capacity is lower in endocrine-resistant cells. • Endocrine-resistant breast cancer cells have increased glycolysis. • Bioenergetic responses to E2 and tamoxifen are lower in endocrine-resistant cells.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2023

Multigenic resistance to Xylella fastidiosa in wild grapes ( Vitis sps.) and its implications within a changing climate

Xylella fastidiosa is a bacterium that infects crops like grapevines, coffee, almonds, citrus and olives. There is little understanding of the genes that contribute to plant resistance, the genomic architecture of resistance, and the potential role of climate in shaping resistance, in part because major crops like grapevines (Vitis vinifera) are not resistant to the bacterium. Here we study a wild grapevine species, V. arizonica, that segregates for resistance. Using genome-wide association, we identify candidate resistance genes. Resistance-associated kmers are shared with a sister species of V. arizonica but not with more distant species, suggesting that resistance evolved more than once. Finally, resistance is climate dependent, because individuals from low (< 10 °C) temperature locations in the wettest quarter were typically susceptible to infection, likely reflecting a lack of pathogen pressure in colder climates. In fact, climate is as effective a predictor of resistance phenotypes as some genetic markers. We extend our climate observations to additional crops, predicting that increased pathogen pressure is more likely for grapevines and almonds than some other susceptible crops.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2026

Recent efforts of vapour-phase strategies for EUV resist toward high- and hyper-NA extreme ultraviolet lithography

Extreme ultraviolet lithography (EUVL, λ = 13.5 nm) is critical for sub-1 nm technology nodes but remains constrained by inherent trade-offs among resolution, line-edge roughness (LER), and sensitivity. Stochastic effects originating from photon shot noise, low-energy secondary electron blur, and the random distribution of resist components further limit its advancement toward high numerical-aperture (NA, NA = 0.55) and hyper-NA (NA ≥0.75) EUVL. While the optimization of spin-on chemically amplified resists (CARs) continues, metal-oxide resists (MORs) have emerged as strong candidates for next-generation EUVL by incorporating metals with high EUV absorption coefficients, which enhances both resist sensitivity and etch resistance during pattern transfer. Besides spin-coating, recent advances in vapour-phase techniques, such as vapour-phase infiltration (VPI), chemical vapour deposition (CVD), and molecular atomic layer deposition (MALD), offer promising pathways to achieve new resist platforms, such as dry resists, that satisfy the stringent thickness and uniformity requirements of next-generation EUVL. These methods enable the direct incorporation of metal species into existing resist matrices or the formation of hybrid inorganic–organic resist platforms, thereby improving film uniformity, etch durability, and pattern fidelity while mitigating stochastic defects. This review highlights the latest advancements in vapour-phase-synthesized EUV resists, emphasizing material design, lithographic performance, and the underlying exposure mechanisms. Although still emerging, vapour-phase strategies are paving the way for an all-dry integration framework that could improve EUV patterning workflows and meet the demands of future technology nodes.

36 MATERIALS SCIENCE↗

DOE OSTI2016

Novel Metal Cation Resistance Systems from Mutant Fitness Analysis of Denitrifying Pseudomonas stutzeri

Metal ion transport systems have been studied extensively, but the specificity of a given transporter is often unclear from amino acid sequence data alone. In this study, predicted Cu 2+ and Zn 2+ resistance systems in Pseudomonas stutzeri strain RCH2 are compared with those experimentally implicated in Cu 2+ and Zn 2+ resistance, as determined by using a DNA-barcoded transposon mutant library. Mutant fitness data obtained under denitrifying conditions are combined with regulon predictions to yield a much more comprehensive picture of Cu 2+ and Zn 2+ resistance in strain RCH2. The results not only considerably expand what is known about well-established metal ion exporters (CzcCBA, CzcD, and CusCBA) and their accessory proteins (CzcI and CusF), they also reveal that isolates with mutations in some predicted Cu 2+ resistance systems do not show decreased fitness relative to the wild type when exposed to Cu 2+ . In addition, new genes are identified that have no known connection to Zn 2+ ( corB , corC , Psest_3226, Psest_3322, and Psest_0618) or Cu 2+ resistance (Mrp antiporter subunit gene, Psest_2850, and Psest_0584) but are crucial for resistance to these metal cations. Growth of individual deletion mutants lacking corB , corC , Psest_3226, or Psest_3322 confirmed the observed Zn-dependent phenotypes. Notably, to our knowledge, this is the first time a bacterial homolog of TMEM165, a human gene responsible for a congenital glycosylation disorder, has been deleted and the resulting strain characterized. Finally, the fitness values indicate Cu 2+ - and Zn 2+ -based inhibition of nitrite reductase and interference with molybdenum cofactor biosynthesis for nitrate reductase. These results extend the current understanding of Cu 2+ and Zn 2+ efflux and resistance and their effects on denitrifying metabolism. Here in this study, genome-wide mutant fitness data in P. stutzeri RCH2 combined with regulon predictions identify several proteins of unknown function that are involved in resisting zinc and copper toxicity. For zinc, these include a member of the UPF0016 protein family that was previously implicated in Ca 2+ /H + antiport and a human congenital glycosylation disorder, CorB and CorC, which were previously linked to Mg 2+ transport, and Psest_3322 and Psest_0618, two proteins with no characterized homologs. Experiments using mutants lacking Psest_3226, Psest_3322, corB , corC , or czcI verified their proposed functions, which will enable future studies of these little-characterized zinc resistance determinants. Likewise, Psest_2850, annotated as an ion antiporter subunit, and the conserved hypothetical protein Psest_0584 are implicated in copper resistance. Physiological connections between previous studies and phenotypes presented here are discussed. Functional and mechanistic understanding of transport proteins improves the understanding of systems in which members of the same protein family, including those in humans, can have different functions.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2019

Comprehensive Genetic Search to Clarify the Molecular Mechanism of Drug Resistance Identifies ASCL2-LEF1/TSPAN8 Axis in Colorectal Cancer

Background: Treatment-resistance genes limiting anticancer therapy have not been well clarified in colorectal cancer (CRC). We explored gene expression profiles to identify biomarkers for predicting treatment resistance to an anticancer drug in CRC. Methods: Six CRC cell lines were treated with phenylbutyrate (PB). The gene expression profiles were then compared using microarrays (harboring 54,675 genes), and genes associated with PB resistance were identified. Candidate genes were functionally examined in cell lines and clinically validated for treatment resistance in clinical samples. Results: Both DLD1 and HCT15 cells were PB resistant, while HCT116 cells were identified as PB sensitive. On microarray analysis, among the PB resistance-related genes, the expression of the genes ASCL2, LEF1, and TSPAN8 was clearly associated with PB resistance. PB-sensitive cells transfected with one of these three genes exhibited significant (P < 0.001) augmentation of PB resistance; ASCL2 induced expression of both LEF1 and TSPAN8, while neither LEF1 nor TSPAN8 induced ASCL2. RNA interference via ASCL2 knockdown made PB-resistant cells sensitive to PB and inhibited both genes. ASCL2 knockdown also played a critical role in sensitivity to treatment by 5-fluorouracil and radiotherapy in addition to PB. Finally, ASCL2 expression was significantly correlated with histological grade of rectal cancer with preoperative chemoradiation therapy. Conclusions: ASCL2 was identified as a causative gene involved in therapeutic resistance against anticancer treatments in CRC.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

DOE OSTI2018

Genome-Wide Association Mapping of Anthracnose ( Colletotrichum sublineolum ) Resistance in the U.S. Sorghum Association Panel

The productivity and profitability of sorghum [Sorghum bicolor (L.) Moench] is reduced by susceptibility to fungal diseases, such as anthracnose (Colletotrichum sublineolum P. Henn.). A limited number of resistant accessions are present in the temperate-adapted germplasm; other exotic sources of resistance are not currently available for breeding programs. Among 335 accessions available to breeders from a previously genotyped sorghum association panel (SAP), we found that 75 were resistant to anthracnose. A phylogenetic analysis of these accessions showed high genetic diversity and multiple resistance sources. Genome-wide association scans (GWAS) were conducted using 268,289 single-nucleotide polymorphisms to identify loci associated with anthracnose resistance. Using logistic regressions for binary measures of resistance responses, we identified three loci within a region on chromosome 5 that have been previously associated with three sources of anthracnose resistance. A GWAS limited to Caudatum germplasm identified an association with a region on chromosome 1 and with the same previous region on chromosome 5. Candidate genes within these loci were related to R-gene families, signaling cascades, and transcriptional reprogramming, suggesting that the resistance response is controlled by multiple defense mechanisms. The strategic integration of exotic resistant germplasm into the SAP is needed to identify additional rare resistance alleles via GWAS.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS1989

Resistivity of pristine and intercalated graphite fiber epoxy composites

Laminar composites were fabricated from pristine and bromine intercalated Amoco P-55, P-75, and P-100 graphite fibers and Hysol-Grafil EAG101-1 film epoxy. The thickness and r.f. eddy current resistivity of several samples were measured at grid points and averaged point by point to obtain final values. Although the values obtained this way have high precision (less than 3 percent deviation), the resistivity values appear to be 20 to 90 percent higher than resistivities measured on high aspect ratio samples using multi-point techniques, and by those predicted by theory. The temperature dependence of the resistivity indicates that the fibers are neither damaged nor deintercalated by the composite fabrication process. The resistivity of the composites is a function of sample thickness (i.e., resin content). Composite resistivity is dominated by fiber resistivity, so lowering the resistivity of the fibers, either through increased graphitization or intercalation, results in a lower composite resistivity. A modification of the simple rule of mixtures model appears to predict the conductivity of high aspect ratio samples measured along a fiber direction, but a directional dependence appears which is not predicted by the theory. The resistivity of these materials is clearly more complex than that of homogeneous materials.

Gaier, James R.↗

NASA NTRS1991

Resistivity of pristine and intercalated graphite fiber epoxy composites

Gaier, James R.↗

NASA NTRS2010

Electrically Variable Resistive Memory Devices

Nonvolatile electronic memory devices that store data in the form of electrical- resistance values, and memory circuits based on such devices, have been invented. These devices and circuits exploit an electrically-variable-resistance phenomenon that occurs in thin films of certain oxides that exhibit the colossal magnetoresistive (CMR) effect. It is worth emphasizing that, as stated in the immediately preceding article, these devices function at room temperature and do not depend on externally applied magnetic fields. A device of this type is basically a thin film resistor: it consists of a thin film of a CMR material located between, and in contact with, two electrical conductors. The application of a short-duration, low-voltage current pulse via the terminals changes the electrical resistance of the film. The amount of the change in resistance depends on the size of the pulse. The direction of change (increase or decrease of resistance) depends on the polarity of the pulse. Hence, a datum can be written (or a prior datum overwritten) in the memory device by applying a pulse of size and polarity tailored to set the resistance at a value that represents a specific numerical value. To read the datum, one applies a smaller pulse - one that is large enough to enable accurate measurement of resistance, but small enough so as not to change the resistance. In writing, the resistance can be set to any value within the dynamic range of the CMR film. Typically, the value would be one of several discrete resistance values that represent logic levels or digits. Because the number of levels can exceed 2, a memory device of this type is not limited to binary data. Like other memory devices, devices of this type can be incorporated into a memory integrated circuit by laying them out on a substrate in rows and columns, along with row and column conductors for electrically addressing them individually or collectively.

Liu, Shangqing↗

NASA NTRS2010

A Constant-Force Resistive Exercise Unit

A constant-force resistive exercise unit (CFREU) has been invented for use in both normal gravitational and microgravitational environments. In comparison with a typical conventional exercise machine, this CFREU weighs less and is less bulky: Whereas weight plates and associated bulky supporting structures are used to generate resistive forces in typical conventional exercise machines, they are not used in this CFREU. Instead, resistive forces are generated in this CFREU by relatively compact, lightweight mechanisms based on constant-torque springs wound on drums. Each such mechanism is contained in a module, denoted a resistive pack, that includes a shaft for making a torque connection to a cable drum. During a stroke of resistive exercise, the cable is withdrawn from the cable drum against the torque exerted by the resistance pack. The CFREU includes a housing, within which can be mounted one or more resistive pack(s). The CFREU also includes mechanisms for engaging any combination of (1) one or more resistive pack(s) and (2) one or more spring(s) within each resistive pack to obtain a desired level of resistance.

Colosky, Paul↗

DOE OSTI2014

Resistive foil edge grading for accelerator and other high voltage structures

In a structure or device having a pair of electrical conductors separated by an insulator across which a voltage is placed, resistive layers are formed around the conductors to force the electric potential within the insulator to distribute more uniformly so as to decrease or eliminate electric field enhancement at the conductor edges. This is done by utilizing the properties of resistive layers to allow the voltage on the electrode to diffuse outwards, reducing the field stress at the conductor edge. Preferably, the resistive layer has a tapered resistivity, with a lower resistivity adjacent to the conductor and a higher resistivity away from the conductor. Generally, a resistive path across the insulator is provided, preferably by providing a resistive region in the bulk of the insulator, with the resistive layer extending over the resistive region.

42 ENGINEERING↗

DOE OSTI2024

Relating flow resistance to equivalent roughness

Describing flow resistance using the physical properties of an underlying surface is a recalcitrant problem in overland flow models. If discharge measurements are available, an equivalent roughness (e.g., Manning’s n) can be calibrated to represent the effects of surface properties within the domain with a single numerical value. Alternatively, the flow resistance can be estimated from discharge and velocity measured at a point, typically a runoff plot outlet. However, such experimental estimates are often inconsistent with the equivalent roughness determined from calibration to discharge, even if both derive from the same dataset. For example, if Manning’s equation is used to parameterize flow resistance, the Manning’s n obtained by calibrating a model to discharge differs from the value of n calculated from measured flow and velocity at the hillslope outlet. Here, this discrepancy is resolved by deriving a correction factor relating experimentally-determined flow resistance to the equivalent roughness. The derived correction factor is tested for four commonly-used resistance formulations using 129 rainfall simulator experiments. The correction factor is necessary to reproduce measured velocities, and yields minor improvements in discharge prediction. Plain Language Summary: Accurate runoff prediction is needed for land and water management in dryland regions, where sporadic and limited rainfall necessitate efficient water use and drought mitigation strategies. The skill of runoff models is known to be hindered by out ability to estimate flow resistance, which is the quantity that describes how energy is lost from flowing water to the underlying surface. Typically, models represent flow resistance with an equivalent roughness, e.g., Manning’s n, that is adjusted until the model can reproduce available discharge observations at watershed scale. However, the flow resistance measured in plot-scale experiments (1–10 m) often exceeds equivalent roughness coefficients by a factor of 10. This means that the direct use of plot-scale experimental data to parameterize runoff models could cause errors in discharge and runoff velocity predictions. Here, we resolve these differences by deriving an analytic correction factor that relates flow resistance to the equivalent roughness required for models to reproduce experimental velocity and discharge data. This correction factor is tested using rainfall simulator data from 129 experiments performed in the US Southwest covering a wide range of precipitation intensities, soil textures and vegetation types. Use of the correction factor substantially improves model prediction of flow velocity, which is needed for reproducing the timing of flood events and the estimation of erosion.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2018

Whole-genome sequencing and single nucleotide polymorphisms in multidrug-resistant clinical isolates of Mycobacterium tuberculosis from the Philippines

Thousands of cases of multidrug-resistant tuberculosis (TB) have been observed in the Philippines, but studies on the Mycobacterium tuberculosis (MTB) genotypes that underlie the observed drug resistance profiles are lacking. This study aimed to analyse the whole genomes of clinical MTB isolates representing various resistance profiles to identify single nucleotide polymorphisms (SNPs) in resistance-associated genes. The genomes of ten MTB isolates cultured from banked sputum sources were sequenced. Bioinformatics analysis consisted of assembly, annotation and SNP identification in genes reported to be associated with resistance to isoniazid (INH), rifampicin (RIF), ethambutol (ETH), streptomycin, pyrazinamide (PZA) and fluoroquinolones (FQs). The draft assemblies covered an average of 97.08% of the expected genome size. Seven of the ten isolates belonged to the Indo-Oceanic lineage/EA12-Manila clade. Two isolates were classified into the Euro-American lineage, whilst the pre-XDR (pre-extensively drug-resistant) isolate was classified under the East Asian/Beijing clade. The SNPs katG Ser315Thr, rpoB Ser450Leu and embB Met306Val were found in INH- (4/7), RIF- (3/6) and ETH-resistant (2/6) isolates, respectively, but not in susceptible isolates. Mutations in the inhA promoter and in the pncA and gyrA genes known to be involved in resistance to INH, PZA and FQs, respectively, were also identified. This study represents the first effort to investigate the whole genomes of Philippine clinical strains of MTB exhibiting various multidrug resistance profiles. Whole-genome data can provide valuable insights to the mechanistic and epidemiological qualities of TB in a high-burden setting such as the Philippines.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2023

Co-regulation of water and energy in the spatial heterogeneity of drought resistance and resilience

Vegetation resistance and resilience to drought are linked to the stability of terrestrial ecosystems under climate change. However, the factors driving the spatial heterogeneity in drought resistance and resilience remain poorly understood. In the study, we utilized multiple satellite-derived vegetation indices to calculate and analyze changes in drought resistance and resilience across various biomes worldwide. Results indicated that drought resistance showed a significant increase with the increase in water availability, but no significant relationship was observed between drought resistance and energy. In contrast, drought resilience exhibited a significant increase with an increase in energy rather than in water. Furthermore, a negative correlation was observed between drought resistance and resilience across different biomes worldwide, indicating a trade-off between resistance and resilience. However, the strength of the negative correlation varied based on water and energy conditions. These findings provide compelling evidence that water and energy co-regulated the spatial heterogeneity in drought resistance and resilience across the globe. The robust linear relationship between drought resistance and resilience and available water and energy demonstrated in our study is critical to accurately predicting and assessing the impact of climate change on vegetation growth and terrestrial carbon cycling in the future.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2022

Li-Ion Battery Electrode Contact Resistance Estimation by Mechanical Peel Test

Li-ion battery electrode electronic properties, including bulk conductivity and contact resistance, are critical parameters affecting cell performance and fast-charge capability. Contact resistance between the coating and current collector is often the largest electronic resistance in an electrode and is affected by chemical, microstructural, and interfacial variations. Direct measurements of contact resistance and bulk conductivity have proven to be challenging. In their absence, a mechanical electrode peel test is often used to compare adhesion and electrical contact resistance. However, using a micro-flexible-surface probe, contact resistance can be directly determined. Here, this work compares contact resistance and mechanical peel strength of multiple commercial-grade HE5050 and NCM523 cathodes and graphite and silicon anodes. It was found that peel strength correlates well with contact resistance in a carefully curated data set (p < 0.05) and in some situations may be a good metric to estimate electrical properties. However, there were distinct outliers in the data set, indicating that peel strength may not accurately reflect electrical properties when there is significant variation in electrode composition. These results illustrate the value of the micro-flexible-surface probe in quantifying contact resistance and bulk conductivity to better understand how battery composition and processing steps affect microstructure and resulting cell performance.

25 ENERGY STORAGE↗

DOE OSTI2023

In silico analyses of diversity and dissemination of antimicrobial resistance genes and mobile genetics elements, for plasmids of enteric pathogens

The antimicrobial resistance (AMR) mobilome plays a key role in the dissemination of resistance genes encoded by mobile genetics elements (MGEs) including plasmids, transposons (Tns), and insertion sequences (ISs). These MGEs contribute to the dissemination of multidrug resistance (MDR) in enteric bacterial pathogens which have been considered as a global public health risk. To further understand the diversity and distribution of AMR genes and MGEs across different plasmid types, we utilized multiple sequence-based computational approaches to evaluate AMR-associated plasmid genetics. A collection of 1,309 complete plasmid sequences from Gammaproteobacterial species, including 100 plasmids from each of the following 14 incompatibility (Inc) types: A/C, BO, FIA, FIB, FIC, FIIA, HI1, HI2, I1, K, M, N, P except W, where only 9 sequences were available, was extracted from the National Center for Biotechnology Information (NCBI) GenBank database using BLAST tools. The extracted FASTA files were analyzed using the AMRFinderPlus web-based tools to detect antimicrobial, disinfectant, biocide, and heavy metal resistance genes and ISFinder to identify IS/Tn MGEs within the plasmid sequences. In silico prediction based on plasmid replicon types showed that the resistance genes were diverse among plasmids, yet multiple genes were widely distributed across the plasmids from enteric bacterial species. These findings provide insights into the diversity of resistance genes and that MGEs mediate potential transmission of these genes across multiple plasmid replicon types. This notion was supported by the observation that many IS/Tn MGEs and resistance genes known to be associated with them were common across multiple different plasmid types. Our results provide critical insights about how the diverse population of resistance genes that are carried by the different plasmid types can allow for the dissemination of AMR across enteric bacteria. The results also highlight the value of computational-based approaches and in silico analyses for the assessment of AMR and MGEs, which are important elements of molecular epidemiology and public health outcomes.

59 BASIC BIOLOGICAL SCIENCES↗