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At least 145 records · Page 8

Developing a pipeline to expand the genetic code of diverse bacteria for microbial engineering

Microbial biotechnologies are key to addressing grand challenges to promote human health, reverse carbon emissions, recycle mixed plastic waste, remediate contaminated soils, and achieve sustainable economies. Synthetic biology has enabled design of diverse microbes and their proteins for useful purposes, but the narrowness of the natural genetic code limits functional diversity (e.g., biosynthesis) of engineered microbes. The natural genetic code defines the fundamental rules of translating genetic information into proteins comprised of 22 ‘canonical’ amino acids. However, using a technique called genetic code expansion (GCE), the chemical properties and therefore functions of proteins can be transformed by incorporation of one or more of ~200 chemically diverse ‘non-canonical’ amino acids. The effective application of genetic code expansion in diverse microbes has the potential to revolutionize biotechnology. However, despite over 50 years of research and its transformative potential, the application of genetic code expansion has been limited to a handful of bacterial species. In this project, we will perform three tasks to both overcome the barriers that prevent wide spread adoption of GCE as molecular tool and demonstrate its potential for biotechnological applications. Specifically, we will (1) develop a genetic engineering methodology that will enable use of GCE in a broad range of bacterial hosts, (2) use high-throughput functional genomics methods to identify physiological responses to both genetic code expansion and exposure to non-canonical amino acids in three different bacteria, and (3) demonstrate an application of GCE by selectively incorporate non-canonical amino acids into surface displayed peptides such as those used for biomining.

59 BASIC BIOLOGICAL SCIENCES↗

Plant Growth Promotion Diversity in Switchgrass-Colonizing, Diazotrophic Endophytes

Endophytic nitrogen-fixing (diazotrophic) bacteria are essential members of the microbiome of switchgrass ( Panicum virgatum ), considered to be an important commodity crop in bioenergy production. While endophytic diazotrophs are known to provide fixed atmospheric nitrogen to their host plant, there are many other plant growth-promoting (PGP) capabilities of these organisms to be demonstrated. The diversity of PGP traits across different taxa of switchgrass-colonizing endophytes is understudied, yet critical for understanding endophytic function and improving cultivation methods of important commodity crops. Here, we present the isolation and characterization of three diazotrophic endophytes: Azospirillum agricola R1C, Klebsiella variicola F10Cl, and Raoultella terrigena R1Gly. Strains R1C and F10Cl were isolated from switchgrass and strain R1Gly, while isolated from tobacco, is demonstrated herein to colonize switchgrass. Each strain exhibited highly diverse genomic and phenotypic PGP capabilities. Strain F10Cl and R1Gly demonstrated the highest functional similarity, suggesting that, while endophyte community structure may vary widely based on host species, differences in functional diversity are not a clearly delineated. The results of this study advance our understanding of diazotrophic endophyte diversity, which will allow us to design robust strategies to improve cultivation methods of many economically important commodity crops.

59 BASIC BIOLOGICAL SCIENCES↗

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↗

Machine-learning based approach to examine ecological processes influencing the diversity of riverine dissolved organic matter composition

Dissolved organic matter (DOM) assemblages in freshwater rivers are formed from mixtures of simple to complex compounds that are highly variable across time and space. These mixtures largely form due to the environmental heterogeneity of river networks and the contribution of diverse allochthonous and autochthonous DOM sources. Most studies are, however, confined to local and regional scales, which precludes an understanding of how these mixtures arise at large, e.g., continental, spatial scales. The processes contributing to these mixtures are also difficult to study because of the complex interactions between various environmental factors and DOM. Here we propose the use of machine learning (ML) approaches to identify ecological processes contributing toward mixtures of DOM at a continental-scale. We related a dataset that characterized the molecular composition of DOM from river water and sediment with Fourier-transform ion cyclotron resonance mass spectrometry to explanatory physicochemical variables such as nutrient concentrations and stable water isotopes ( 2 H and 18 O). Using unsupervised ML, distinctive clusters for sediment and water samples were identified, with unique molecular compositions influenced by environmental factors like terrestrial input and microbial activity. Sediment clusters showed a higher proportion of protein-like and unclassified compounds than water clusters, while water clusters exhibited a more diversified chemical composition. We then applied a supervised ML approach, involving a two-stage use of SHapley Additive exPlanations (SHAP) values. In the first stage, SHAP values were obtained and used to identify key physicochemical variables. These parameters were employed to train models using both the default and subsequently tuned hyperparameters of the Histogram-based Gradient Boosting (HGB) algorithm. The supervised ML approach, using HGB and SHAP values, highlighted complex relationships between environmental factors and DOM diversity, in particular the existence of dams upstream, precipitation events, and other watershed characteristics were important in predicting higher chemical diversity in DOM. Our data-driven approach can now be used more generally to reveal the interplay between physical, chemical, and biological factors in determining the diversity of DOM in other ecosystems.

54 ENVIRONMENTAL SCIENCES↗

Genetic and Structural Diversity of Prokaryotic Ice-Binding Proteins from the Central Arctic Ocean.

Ice-binding proteins (IBPs) are a group of ecologically and biotechnologically relevant enzymes produced by psychrophilic organisms. Although putative IBPs containing the domain of unknown function (DUF) 3494 have been identified in many taxa of polar microbes, our knowledge of their genetic and structural diversity in natural microbial communities is limited. Here, we used samples from sea ice and sea water collected in the central Arctic Ocean as part of the MOSAiC expedition for metagenome sequencing and the subsequent analyses of metagenome-assembled genomes (MAGs). By linking structurally diverse IBPs to particular environments and potential functions, we reveal that IBP sequences are enriched in interior ice, have diverse genomic contexts and cluster taxonomically. Their diverse protein structures may be a consequence of domain shuffling, leading to variable combinations of protein domains in IBPs and probably reflecting the functional versatility required to thrive in the extreme and variable environment of the central Arctic Ocean.

59 BASIC BIOLOGICAL SCIENCES↗

Soil carbon accrual and biopore formation across a plant diversity gradient

Plant diversity promotes soil organic carbon (SOC) gains through intricate changes in root-soil interactions and their subsequent influence on soil physical and biological processes. We assessed SOC and pore characteristics of soils under a range of switchgrass-based plant systems 12 years after their establishment. The systems represented a gradient of plant diversity with species richness ranging from 1 to 30 species. We focused on soil biopores as indicators of the legacy of root activity and explored biopore relationships with SOC accumulation. Biopores were measured using X-ray computed micro-tomography. Plant functional richness explained 29 % of bioporosity and 36 % of SOC variation, while bioporosity itself explained 36 % of the variation in SOC. The most diverse plant system (30 species) had the highest SOC, while long-term bare soil fallow and monoculture switchgrass had the lowest. Of particular note was a 2-species mixture of switchgrass (Panicum virgatum L.) and ryegrass (Elymus canadensis), which exhibited the highest bioporosity and achieved SOC levels comparable to those of the systems with 6 and 10 plant species, and were inferior only to the system with 30 species. We conclude that plant diversity may enhance SOC through biopore-mediated mechanisms and suggest a potential for identifying specific plant combinations that may be particularly efficient for fostering biopore formation and, subsequently, SOC sequestration.

Kim, Kyungmin [Seoul National Univ. (Korea, Republ↗

Initial results from the VPI&SU SIRIO diversity experiment

The first year of observations of the precipitation effects on the 11.6-GHz beacon signal from the SIRIO satellite are reported. This experiment is unique in that it uses dual-polarized receivers in a diversity configuration and a low path elevation angle. Rain rate, attenuation, and isolation statistics are presented for both sites. In addition, attenuation diversity gain is plotted and compared to current models. Isolation diversity gain is also discussed. It was found that little improvement in dual-channel isolation is obtained with site diversity at this low elevation angle. The influence of ice clouds was noted in the months of October and November.

Towner, G. C.↗

Coherent lidar signal fluctuation reduction by means of frequency diversity technique

The atmospheric return measured by a coherent lidar is typically characterized by rapid and deep fluctuations in signal strength. These fluctuations result from the interference of the fields backscattered to the lidar from randomly located aerosol particles which move relative to the lidar pulse. In many applications, it is necessary to determine the average value of the lidar signal intensity at some range. A new method utilizes frequency diversity initially suggested by Goldstein and subsequently studied in the microwave radar domain by others. It is expected that the application of the frequency diversity method in the coherent lidar domain will eventually provide greater efficiency and speed in the return signal averaging needed to obtain accurate intensity estimates. The frequency diversity method recognizes that the transmitted lidar pulse is very long compared to a wavelength and consequently a given phase, theta sub i, is repeated many times within the pulse. In order to test this concept, a fairly simple laboratory experiment was designed which simulates scattering of a lidar pulse from atmospheric aerosol. The testing of the frequency diversity method is discussed.

Schotland, R. M.↗

Endangered and potentially endangered wildlife on John F. Kennedy Space Center and faunal integrity as a goal for maintaining biological diversity

Buffer zones for space operations provide for a wildlife diversity unsurpassed among most federal facilities in the continental U.S. demonstrating the coexistence possible with one of man's greatest technological achievements. This document ranks 119 resident or migratory wildlife species that are endangered or declining. The ranking system herein was based on species' vulnerability to extinction and the relevance of Kennedy Space Center (KSC) for maintaining populations in the U.S. and Florida. One amphibian, 19 reptiles, 80 birds, and 19 mammals were considered endangered or declining. KSC is an integral area for regional species diversity being the focus of the Merritt Island/Cape Canaveral/Turnbull Ecosystem which is part of the Indian River Lagoon watershed, an estuary of national significance. Many species that use this system also use the nearby St. Johns River Basin ecosystem. These two ecosystems are biological corridors between temperate Carolinian and tropical/subtropical Caribbean biotic provinces. Threats to biological diversity on KSC were also reviewed. Traditional environmental assessments, resulting from environmental regulation guidelines, focus on environmental contaminants and habitat lost due to construction. However, this review suggested that small population sizes, isolation of populations, ecosystem and habitat fragmentation, road mortality, and other edge effects may represent more critical threats to biological diversity than the traditional topics.

Breininger, David R.↗

Dissecting global diversity patterns: examples from the Ordovician Radiation

Although the history of life has been characterized by intermittent episodes of radiation that can be recognized in global compilations of biodiversity, it does not necessarily follow that these episodes are caused by processes that occurred uniformly around the world. Major diversity increases could be generated by the cumulative effects of different mechanisms operating simultaneously at several geographic or environmental scales. The purpose of this review is to describe ongoing research on the manifestations, at several scales, of the Ordovician Radiation, which was among the most extensive intervals of diversification in the history of life. Through much of the period, diversity was concentrated most heavily near regions of active mountain building and volcanism; differences in diversity patterns from continent to continent, and among regions within continents, reflect this overprint. While this suggests a linkage of the Radiation and tectonic activity, this is by no means the only mediating agent. Outcrop-based research in North America has demonstrated that tectonic activity was detrimental to some biotic elements, in contrast to its effects on other organisms. Moreover, in the Great Basin of North America where the local stratigraphic record is of particularly high quality, biotic transitions characteristic of the period occurred far more rapidly than observed in global compilations of diversity, suggesting that the global rate of transition may represent the aggregate sum of transitions that occurred abruptly, but at different times, around the world. Finally, it has been demonstrated that, in concert with an increase in average age, the environmental and geographic ranges of Ordovician genera both increased significantly through the period, indicating a role for intrinsic factors in producing Ordovician biotic patterns.

Non-NASA Center↗

Effects of posture on blood flow diversion by hypoxic pulmonary vasoconstriction in dogs

We used differential excretion of sulphur hexafluoride from the left and right lung to measure blood flow diversion by hypoxic pulmonary vasoconstriction (HPV) in the prone and supine positions in dogs (n = 9). Gas exchange was assessed using the multiple inert gas elimination technique. Blood flow diversion from the hypoxic (3% oxygen) left lung was mean 70.7 (SD 11.2)% in the supine compared with 57.0 (12.1)% in the prone position (P < 0.02). The supine position was associated with increased perfusion to low VA/Q regions (P < 0.05). The increased flow diversion with hypoxia in the supine position was associated with more ventilation to high VA/Q regions (P < 0.05). We conclude that flow diversion by hypoxic pulmonary vasoconstriction is greater in the supine position. This effect could contribute to the variable response in gas exchange with positioning in patients with ARDS.

Non-NASA Center↗

Diversity of form in the amphibian papilla of Puerto Rican frogs

In modern frogs, the amphibian papilla exhibits a caudal extension whose shape, relative length, and proportion of hair cells vary markedly from species to species. Tuning in the caudal extension is organized tonotopically and evidently involves the tectorium. In terms of the proportion of amphibian-papillar hair cells in the caudal extension, we report more diversity among 8 species of a single genus (Eleutherodactylus) on a single island (Puerto Rico) than has been found so far among all of the (more than 50) other modern anurans examined for this feature from around the world. These 8 Puerto Rican species have overlapping habitat and conspicuous diversity in the male advertisement call. For 7 of the 8 species, we report that the call has transient spectral components in the frequency range of the amphibian papilla, and that the proportion of caudal extension hair cells and the frequency distribution of those components are correlated. Thus one might conclude that the selective pressures that led to diversity of calls among the 8 species also led to diversity in form of the amphibian papilla.

Non-NASA Center↗

Comparison of the Microbial Diversity and Abundance Between the Freshwater Land-Locked Lakes of Schirmacher Oasis and the Perennially Ice-Covered Lake Untersee in East Antarctica

Extreme conditions such as low temperature, dryness, and constant UV-radiation in terrestrial Antarctica are limiting factors of the survival of microbial populations. The objective of this study was to investigate the microbial diversity and enumeration between the open water lakes of Schirmacher Oasis and the permanently ice-covered Lake Untersee. The lakes in Schirmacher Oasis possessed abundant and diverse group of microorganisms compared to the Lake Untersee. Furthermore, the microbial diversity between two lakes in Schirmacher Oasis (Lake L27C and L47) was compared by culture-based molecular approach. It was determined that L27Chad a richer microbial diversity representing 5 different phyla and 7 different genera. In contrast L47 consisted of 4 different phyla and 6 different genera. The difference in microbial community could be due to the wide range of pH between L27C (pH 9.1) and L47 (pH 5.7). Most of the microbes isolated from these lakes consisted of adaptive biological pigmentation. Characterization of the microbial community found in the freshwater lakes of East Antarctica is important because it gives a further glimpse into the adaptation and survival strategies found in extreme conditions.

Huang, Jonathan↗

Comet Dust: The Diversity of "Primitive" Particles and Implications

Comet dust is primitive and shows significant diversity. Our knowledge of the properties of primitive particles has expanded significantly through microscale investigations of cosmic dust samples ( IDP's(Interplanetary Dust Particles) and AMM's (Antarctic Micrometeorites)) and of comet dust samples (Stardust and Rosetta's COSIMA), as well as through remote sensing (spectroscopy and imaging) via Spitzer and via spacecraft encounters with 103P/Hartley 2 and 67P/Churyumov-Gerasimenko. Microscale investigations show that comet dust and cosmic dust are particles of unequilibrated materials, including aggregates of materials unequilibrated at submicron scales. We call unequilibrated materials "primitive" and we deduce they were incorporated into ice-rich (H2O-, CO2-, and CO-ice) parent bodies that remained cold, i.e., into comets, because of the lack of aqueous or thermal alteration since particle aggregation; yet some Stardust olivines suggest mild thermal metamorphism. Primitive particles exhibit a diverse range of: structure and typology; size and size distribution of constituents; concentration and form of carbonaceous and organic matter; D-, N-, and O- isotopic enhancements over solar; Mg-, Fe-contents of the silicate minerals; the compositions and concentrations of sulfides, and of less abundant mineral species such as chondrules, CAIs and carbonates. The uniformity within a group of samples points to: aerodynamic sorting of particles and/or particle constituents; the inclusion of a limited range of oxygen fugacities; the inclusion or exclusion of chondrules; a selection of organics. The properties of primitive particles imply there were disk processes that resulted in different comets having particular selections of primitive materials. The diversity of primitive particles has implications for the diversity of materials in the protoplanetary disk present at the time and in the region where the comets formed.

Wooden, Diane H.↗

Braving Diversity

In 2020, staff at the Lunar and Planetary Institute (LPI) initiated discussions on diversity and inclusion topicssuch as microaggressions, power imbalances, and personal identity, with the goal of nurturing and sustaining an inclusive workplace climate. Authors Shupla, Filiberto, Rivera-Valentín, and Svambera determined that rather than conducting a single workshop, holding a series of seminars that they titled “Braving Diversity” on assorted topics would be more meaningful. These seminar and discussion sessions are intentionally designed to supplement, not replace, traditional trainings and workshops on unconscious bias, bystander intervention, harassment, and other topics.Braving Diversity is a series of LPI staff discussions, coordinated by and open to all staff on the challenges related to maintaining an inclusive and accessible environment. The vision is to hold these hour-long sessions, which include current research on diversity and inclusion and facilitated dialogue, several times each year.

C. Shupla↗

Hydraulic Diversity of Forests Regulates Ecosystem Resilience During Drought

Plants influence the atmosphere through fluxes of carbon, water and energy, and can intensify drought through land–atmosphere feedback effects. The diversity of plant functional traits in forests, especially physiological traits related to water (hydraulic) transport, may have a critical role in land–atmosphere feedback, particularly during drought. Here we combine 352 site-years of eddy covariance measurements from 40 forest sites, remote-sensing observations of plant water content and plant functional-trait data to test whether the diversity in plant traits affects the response of the ecosystem to drought. We find evidence that higher hydraulic diversity buffers variation in ecosystem flux during dry periods across temperate and boreal forests. Hydraulic traits were the predominant significant predictors of cross-site patterns in drought response. By contrast, standard leaf and wood traits, such as specific leaf area and wood density, had little explanatory power. Our results demonstrate that diversity in the hydraulic traits of trees mediates ecosystem resilience to drought and is likely to have an important role in future ecosystem–atmosphere feedback effects in a changing climate.

Climate and Earth system modelling↗

Diversity and Inclusion in Spacecraft Science Teams: What Do We Know and What Can We Do About It?

Introduction: Not only does the planetary science community lack diversity [1-3], the subset of the community that participates on spacecraft science team is even less diverse than the community as a whole [1, 4]. Results of 2020 Workforce Survey: Previous studies of the diversity of members of spacecraft science teams made incorrect assumptions about the nature of the data before collecting the data. Those analyses assumed a binary gender and ignored the existence of planetary scientists who are neither men nor women [4-6]. We present here results where demographic data was collected without assumptions; each individual surveyed supplied their own answers to demographic questions. The April 2020 survey of Planetary Scientists, which was conducted by the Statistical Research Center of the American Institute of Physics (AIP) and funded by the American Astronomical Society (AAS)’s Division of Planetary Science (DPS) asked participants their gender with 4 possible responses: Woman, Man, Another identify (please specify if you wish), and Prefer not to answer. 32% of respondents chose Woman, 67% chose Man and 1% chose Another gender identity [1]. The survey also asked demographic questions on race, ethnicity, LGBTQ+ identity, and disability. For a full list of questions, see https://dps.aas.org/sites/dps.aas.org/files/reports/2020/survey2020_questionnaire.pdf. In addition to demographic questions, the 2020 Workforce survey asked how many times respondents had been involved in Mission proposals as a Principal Investigator (PI) and, separately, as a Co-Investigator (CoI) [1]. Answers to questions about mission involvement were correlated with answers to demographic questions and the results show that members of historically underrepresented groups (non-white scientists, women, members of the LGBTQ+ community, and disabled scientists) were less likely to be involved in spacecraft mission proposals than were members of historically overrepresented groups [1]. The figures below show the correlated responses for four different axes of underrepresentation [1]. Note that while the figure on gender shows only Women and Men (due to the small percentage of folks answering “Another gender”), non-binary respondents are included in the LGBTQ+ community figure. Conclusion: Being part of a spacecraft science team is a goal for many planetary scientists. With it comes brand new data, more stable funding, and a sense of awe and exploration. It can lead to a cascade of opportunities from conference and public presentations, to membership in subsequent mission teams, and prestige in the community [4]. As a result, participation in spacecraft teams can be used a measure of success within the field. From the survey results, we see that members of historically excluded groups, even after they have overcome barriers to participating in the field, are still experiencing barriers to success within the field itself. Why?: The diminishing percentage of members of underrepresented groups as a career progresses has been referred to as a “leaky pipeline”. However, this fails to adequately capture the experiences of the members of these underrepresented groups as it implies a passive process. In order to capture the active processes (bias, discrimination, harassment, and other exclusionary behaviors) that contribute to low retention in the workforce, the term “Hostile Obstacle Course” is more useful [7,8]. It is these processes that need to be addressed in order to retain valued members of our community. Moving Forward: In order to broaden participation in planetary science, particularly mission science teams, we need to address conditions that create hostile workplace climates. What can mission teams and other groups do to address these conditions? First, each group/team needs to evaluate their own members to determine what specific barriers exist in their own interactions. One tool to accomplish this would be an anonymous survey designed to understand how team members feel about working within the group. Working with professionals who know how to create and analyze such surveys (often called “climate surveys” when applied to University students, for example) would ensure that the survey meets its goals and does not make assumptions that counter the meaningfulness of the results. Such professionals in EDIA (Equity, Diversity, Inclusion, and Accessibility) and workplace culture can make suggestions for policy changes that would eliminate hostile workplace conditions. Policy changes that are often suggested include instituting professional EDIA training for the team and/or for team leadership, instituting and following a code of conduct [9], including more interactive group activities in group meetings, etc. Training and information on EDIA is available for all members of the planetary science community. The first place to look would be in your University or Institution’s EDIA or human resources offices. Bystander Intervention is often offered as part of other meetings [10]. A newer offering is a Workshop on EDIA for Leaders in Planetary Science led by Julie Rathbun (first author of this abstract) and JA Grier (https://edialps.psi.edu/). This 3-day workshop gives participants the tools they need to enact positive change in their personal and professional spheres. The first workshop was help in November 2022 and another workshop will take place in the late spring 2023 with exact dates to be announced soon.

J. A. Rathbun↗

Data-Driven Load Diversity and Variability Modeling for Quasi-Static Time-Series Simulation on Distribution Feeders

This paper presents a data-driven load modeling methodology for distribution system quasi-static time-series (QSTS) simulation considering both diversity and variability characteristics of distribution loads. Based on our previous work in [1]-[2], a variability library and diversity library have been established based on the realistic high-resolution data collected from actual utility feeders. Given the load profile for the start-of-circuit load of a feeder, the loads on the feeder nodes can be modeled with both diversity and variability instead of being directly scaled from the substation load profile according to the distribution allocation factors. With diversified load models, the load-induced impact on the feeder operation characteristics, such as voltage ramp and regulator operations, can be better considered in QSTS simulation. The proposed modeling methodology has been tested on both the IEEE 123-bus feeder and an actual utility feeder model, and the simulation results have demonstrated the merits of deploying the proposed load modeling methodology.

24 POWER TRANSMISSION AND DISTRIBUTION↗