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At least 1,207 records · Page 67

Dose optimization of an adjuvanted peptide-based personalized neoantigen melanoma vaccine

The advancements in next-generation sequencing have made it possible to effectively detect somatic mutations, which has led to the development of personalized neoantigen cancer vaccines that are tailored to the unique variants found in a patient’s cancer. These vaccines can provide significant clinical benefit by leveraging the patient’s immune response to eliminate malignant cells. However, determining the optimal vaccine dose for each patient is a challenge due to the heterogeneity of tumors. To address this challenge, we formulate a mathematical dose optimization problem based on a previous mathematical model that encompasses the immune response cascade produced by the vaccine in a patient. We propose an optimization approach to identify the optimal personalized vaccine doses, considering a fixed vaccination schedule, while simultaneously minimizing the overall number of tumor and activated T cells. To validate our approach, we perform in silico experiments on six real-world clinical trial patients with advanced melanoma. We compare the results of applying an optimal vaccine dose to those of a suboptimal dose (the dose used in the clinical trial and its deviations). Our simulations reveal that an optimal vaccine regimen of higher initial doses and lower final doses may lead to a reduction in tumor size for certain patients. Our mathematical dose optimization offers a promising approach to determining an optimal vaccine dose for each patient and improving clinical outcomes.

59 BASIC BIOLOGICAL SCIENCES↗

teemi: An open-source literate programming approach for iterative design-build-test-learn cycles in bioengineering

Synthetic biology dictates the data-driven engineering of biocatalysis, cellular functions, and organism behavior. Integral to synthetic biology is the aspiration to efficiently find, access, interoperate, and reuse high-quality data on genotype-phenotype relationships of native and engineered biosystems under FAIR principles, and from this facilitate forward-engineering strategies. However, biology is complex at the regulatory level, and noisy at the operational level, thus necessitating systematic and diligent data handling at all levels of the design, build, and test phases in order to maximize learning in the iterative design-build-test-learn engineering cycle. To enable user-friendly simulation, organization, and guidance for the engineering of biosystems, we have developed an open-source python-based computer-aided design and analysis platform operating under a literate programming user-interface hosted on Github. The platform is called teemi and is fully compliant with FAIR principles. In this study we apply teemi for i) designing and simulating bioengineering, ii) integrating and analyzing multivariate datasets, and iii) machine-learning for predictive engineering of metabolic pathway designs for production of a key precursor to medicinal alkaloids in yeast. The teemi platform is publicly available at PyPi and GitHub.

59 BASIC BIOLOGICAL SCIENCES↗

Changing temperature profiles and the risk of dengue outbreaks

As temperatures change worldwide, the pattern and competency of disease vectors will change, altering the global distribution of both the burden of infectious disease and the risk of the emergence of those diseases into new regions. To evaluate the risk of potential summer dengue outbreaks triggered by infected travelers under various climate scenarios, we develop an SEIR-type model, run numerical simulations, and conduct sensitivity analyses under a range of temperature profiles. Our model extends existing theoretical frameworks for studying dengue dynamics by introducing temperature dependence of two key parameters: the mosquito extrinsic incubation period and the lifespan of mosquitoes, which empirical data suggests are both highly temperature dependent. We find that changing temperature significantly alters dengue risk in an inverted U-shape, with temperatures in the range 27-31°C producing the highest risk. As temperatures increase beyond 31°C, the determinants of dengue risk begin to shift from mosquito biting rate and carrying capacity to the duration of the human infectious period, suggesting that changing temperatures not only alter dengue risk but also the potential efficacy of control measures. To illustrate the role of spatial and temporal temperature heterogeneity, we select five US cities where the primary dengue vector, the mosquito Aedes aegypti , has been observed, and which have had dengue cases in the past: Los Angeles, Houston, Miami, Brownsville, and Phoenix. Our analysis suggests that an increase of 3°C leads to an approximate doubling of the risk of dengue in Los Angeles and Houston, but a reduction of risk in Miami, Brownsville, and Phoenix due to extreme heat.

54 ENVIRONMENTAL SCIENCES↗

Detecting multiple communities using quantum annealing on the D-Wave system

A very important problem in combinatorial optimization is the partitioning of a network into communities of densely connected nodes; where the connectivity between nodes inside a particular community is large compared to the connectivity between nodes belonging to different ones. This problem is known as community detection, and has become very important in various fields of science including chemistry, biology and social sciences. The problem of community detection is a twofold problem that consists of determining the number of communities and, at the same time, finding those communities. This drastically increases the solution space for heuristics to work on, compared to traditional graph partitioning problems. In many of the scientific domains in which graphs are used, there is the need to have the ability to partition a graph into communities with the “highest quality” possible since the presence of even small isolated communities can become crucial to explain a particular phenomenon. We have explored community detection using the power of quantum annealers, and in particular the D-Wave 2X and 2000Q machines. It turns out that the problem of detecting at most two communities naturally fits into the architecture of a quantum annealer with almost no need of reformulation. This paper addresses a systematic study of detecting two or more communities in a network using a quantum annealer.

97 MATHEMATICS AND COMPUTING↗

Duckweed hosts a taxonomically similar bacterial assemblage as the terrestrial leaf microbiome

Culture-independent characterization of microbial communities associated with popular plant model systems have increased our understanding of the plant microbiome. However, the integration of other model systems, such as duckweed, could facilitate our understanding of plant microbiota assembly and evolution. Duckweeds are floating aquatic plants with many characteristics, including small size and reduced plant architecture, that suggest their use as a facile model system for plant microbiome studies. Here, we investigated the structure and assembly of the duckweed bacterial microbiome. First, a culture-independent survey of the duckweed bacterial microbiome from different locations in New Jersey revealed similar phylogenetic profiles. These studies showed that Proteobacteria is a dominant phylum in the duckweed bacterial microbiome. To observe the assembly dynamics of the duckweed bacterial community, we inoculated quasi-gnotobiotic duckweed with wastewater effluent from a municipal wastewater treatment plant. Our results revealed that duckweed strongly shapes its bacterial microbiome and forms distinct associations with bacterial community members from the initial inoculum. Additionally, these inoculation studies showed the bacterial communities of different duckweed species were similar in taxa composition and abundance. Analysis across the different duckweed bacterial communities collected in this study identified a set of “core” bacterial taxa consistently present on duckweed irrespective of the locale and context. Furthermore, comparison of the duckweed bacterial community to that of rice and Arabidopsis revealed a conserved taxonomic structure between the duckweed microbiome and the terrestrial leaf microbiome. Our results suggest that duckweeds utilize similar bacterial community assembly principles as those found in terrestrial plants and indicate a highly conserved structuring effect of leaf tissue on the plant microbiome.

54 ENVIRONMENTAL SCIENCES↗

Using mixed methods to construct and analyze a participatory agent-based model of a complex Zimbabwean agro-pastoral system

Complex social-ecological systems can be difficult to study and manage. Simulation models can facilitate exploration of system behavior under novel conditions, and participatory modeling can involve stakeholders in developing appropriate management processes. Participatory modeling already typically involves qualitative structural validation of models with stakeholders, but with increased data and more sophisticated models, quantitative behavioral validation may be possible as well. In this study, we created a novel agent-basedmodel applied to a specific context: Zimbabwean non-governmental organization the Muonde Trust has been collecting data on their agro-pastoral system for the last 35 years and had concerns about land-use planning and the effectiveness of management interventions in the face of climate change. We collaboratively created an agent-based model of their system using their data archive, qualitatively calibrating it to the observed behavior of the real system without tuning any parameters to match specific quantitative outputs. We then behaviorally validated the model using quantitative community-based data and conducted a sensitivity analysis to determine the relative impact of underlying parameter assumptions, Indigenous management interventions, and different rainfall variation scenarios. We found that our process resulted in a model which was successfully structurally validated and sufficiently realistic to be useful for Muonde researchers as a discussion tool. The model was inconsistently behaviorally validated, however, with some model variables matching field data better than others. We observed increased model system instability due to increasing variability in underlying drivers (rainfall), and also due to management interventions that broke feedbacks between the components of the system. Interventions that smoothed year-to-year variation rather than exaggerating it tended to improve sustainability. The Muonde trust has used the model to successfully advocate to local leaders for changes in land-use planning policy that will increase the sustainability of their system.

54 ENVIRONMENTAL SCIENCES↗

Managing flowering time in Miscanthus and sugarcane to facilitate intra- and intergeneric crosses

Miscanthus is a close relative of Saccharum and a potentially valuable genetic resource for improving sugarcane. Differences in flowering time within and between Miscanthus and Saccharum hinders intra- and interspecific hybridizations. A series of greenhouse experiments were conducted over three years to determine how to synchronize flowering time of Saccharum and Miscanthus genotypes. We found that day length was an important factor influencing when Miscanthus and Saccharum flowered. Sugarcane could be induced to flower in a central Illinois greenhouse using supplemental lighting to reduce the rate at which days shortened during the autumn and winter to 1 min d -1 , which allowed us to synchronize the flowering of some sugarcane genotypes with Miscanthus genotypes primarily from low latitudes. In a complementary growth chamber experiment, we evaluated 33 Miscanthus genotypes, including 28 M . sinensis , 2 M . floridulus , and 3 M . ×giganteus collected from 20.9° S to 44.9° N for response to three day lengths (10 h, 12.5 h, and 15 h). High latitude-adapted M . sinensis flowered mainly under 15 h days, but unexpectedly, short days resulted in short, stocky plants that did not flower; in some cases, flag leaves developed under short days but heading did not occur. In contrast, for M . sinensis and M . floridulus from low latitudes, shorter day lengths typically resulted in earlier flowering, and for some low latitude genotypes, 15 h days resulted in no flowering. However, the highest ratio of reproductive shoots to total number of culms was typically observed for 12.5 h or 15 h days. Latitude of origin was significantly associated with culm length, and the shorter the days, the stronger the relationship. Nearly all entries achieved maximal culm length under the 15 h treatment, but the nearer to the equator an accession originated, the less of a difference in culm length between the short-day treatments and the 15 h day treatment. Under short days, short culms for high-latitude accessions was achieved by different physiological mechanisms for M . sinensis genetic groups from the mainland in comparison to those from Japan; for mainland accessions, the mechanism was reduced internode length, whereas for Japanese accessions the phyllochron under short days was greater than under long days. Thus, for M . sinensis , short days typically hastened floral induction, consistent with the expectations for a facultative short-day plant. However, for high latitude accessions of M . sinensis , days less than 12.5 h also signaled that plants should prepare for winter by producing many short culms with limited elongation and development; moreover, this response was also epistatic to flowering. Thus, to flower M . sinensis that originates from high latitudes synchronously with sugarcane, the former needs day lengths >12.5 h (perhaps as high as 15 h), whereas that the latter needs day lengths <12.5 h.

54 ENVIRONMENTAL SCIENCES↗

A robust method of nuclei isolation for single-cell RNA sequencing of solid tissues from the plant genus Populus

Single-cell transcriptome analysis has been extensively applied in humans and animal models to uncover gene expression heterogeneity between the different cell types of a tissue or an organ. It demonstrated its capability to discover key regulatory elements that determine cell fate during developmental programs. Single-cell analysis requires the isolation and labeling of the messenger RNA (mRNA) derived from each cell. These challenges were primarily addressed in mammals by developing microfluidic-based approaches. For plant species whose cells contain cell walls, these approaches have generally required the generation of isolated protoplasts. Many plant tissues’ secondary cell wall hinders enzymatic digestion required for individual protoplast isolation, resulting in an unequal representation of cell types in a protoplast population. This limitation is especially critical for cell types located in the inner layers of a tissue or the inner tissues of an organ. Consequently, single-cell RNA sequencing (scRNA-seq) studies using microfluidic approaches in plants have mainly been restricted to Arabidopsis roots, for which well-established procedures of protoplast isolation are available. Here we present a simple alternative approach to generating high-quality protoplasts from plant tissue by characterizing the mRNA extracted from individual nuclei instead of whole cells. We developed the protocol using two different plant materials with varying cellular complexity levels and cell wall structure, Populus shoot apices, and more lignified stems. Using the 10× Genomics Chromium technology, we show that this procedure results in intact mRNA isolation and limited leakage, with a broad representation of individual cell transcriptomes.

54 ENVIRONMENTAL SCIENCES↗

Topographical shifts in fine root lifespan in a mixed, mesic temperate forest

Root lifespan, often is estimated in landscape- and ecosystem-level carbon models using linear approximations. In water manipulation experiments, fine root lifespan can vary with soil water content. Soil water content is generally structured by complex topography, which is largely unaccounted for in landscape- and ecosystem-scale carbon models. Topography governs the range of soil water content experienced by roots which may impact their lifespan. We hypothesized that root lifespan varied nonlinearly across a temperate, mesic, forested catchment due to differences in soil water content associated with topographic position. We expected regions of the landscape that were too wet or too dry would have soils that were not optimal for roots and thus result in shorter root lifespans. Specifically, we hypothesized that root lifespan would be longest in areas that consistently had soil water content in the middle of the soil water content spectrum, while in soils at either very low or very high soil water content, root lifespan would be relatively short. We tested this hypothesis by collecting and analyzing two years of minirhizotron and soil moisture data in plots widely distributed in the Shale Hills catchment of the Susquehanna-Shale Hills Critical Zone Observatory in Pennsylvania. We found that fine root lifespans were longer in traditionally wetter topographic regions, but detected no short term (biweekly) effect of soil moisture on root lifespan. Additionally, depth in soil, soil series, slope face orientation, and season of birth strongly affected root lifespans across the catchment. In contrast, lifespan was unaffected by root diameter or mycorrhizal association. Failure to account for these variables could result in erroneous estimates of fine root lifespan and, consequentially, carbon flux in temperate forested regions.

54 ENVIRONMENTAL SCIENCES↗

Intraspecific variability in plant and soil chemical properties in a common garden plantation of the energy crop Populus

Optimizing crops for synergistic soil carbon (C) sequestration can enhance CO 2 removal in food and bioenergy production systems. Yet, in bioenergy systems, we lack an understanding of how intraspecies variation in plant traits correlates with variation in soil biogeochemistry. This knowledge gap is exacerbated by both the heterogeneity and difficulty of measuring belowground traits. Here, we provide initial observations of C and nutrients in soil and root and stem tissues from a common garden field site of diverse, natural variant, Populus trichocarpa genotypes—established for aboveground biomass-to-biofuels research. Our goal was to explore the value of such field sites for evaluating genotype-specific effects on soil C, which ultimately informs the potential for optimizing bioenergy systems for both aboveground productivity and belowground C storage. To do this, we investigated variation in chemical traits at the scale of individual trees and genotypes and we explored correlations among stem, root, and soil samples. We observed substantial variation in soil chemical properties at the scale of individual trees and specific genotypes. While correlations among elements were observed both within and among sample types (soil, stem, root), above-belowground correlations were generally poor. We did not observe genotype-specific patterns in soil C in the top 10 cm, but we did observe genotype associations with soil acid-base chemistry (soil pH and base cations) and bulk density. Finally, a specific phenotype of interest (high vs low lignin) was unrelated to soil biogeochemistry. Our pilot study supports the usefulness of decade-old, genetically-variable, Populus bioenergy field test plots for understanding plant genotype effects on soil properties. Finally, this study contributes to the advancement of sampling methods and baseline data for Populus systems in the Pacific Northwest, USA. Further species- and region-specific efforts will enhance C predictability across scales in bioenergy systems and, ultimately, accelerate the identification of genotypes that optimize yield and carbon storage.

54 ENVIRONMENTAL SCIENCES↗

Cristae formation is a mechanical buckling event controlled by the inner mitochondrial membrane lipidome

Cristae are high–curvature structures in the inner mitochondrial membrane (IMM) that are crucial for ATP production. While cristae–shaping proteins have been defined, analogous lipid–based mechanisms have yet to be elucidated. Here, we combine experimental lipidome dissection with multi–scale modeling to investigate how lipid interactions dictate IMM morphology and ATP generation. When modulating phospholipid (PL) saturation in engineered yeast strains, we observed a surprisingly abrupt breakpoint in IMM topology driven by a continuous loss of ATP synthase organization at cristae ridges. We found that cardiolipin (CL) specifically buffers the inner mitochondrial membrane against curvature loss, an effect that is independent of ATP synthase dimerization. To explain this interaction, we developed a continuum model for cristae tubule formation that integrates both lipid and protein–mediated curvatures. This model highlighted a snapthrough instability, which drives IMM collapse upon small changes in membrane properties. We also showed that cardiolipin is essential in low–oxygen conditions that promote PL saturation. These results demonstrate that the mechanical function of cardiolipin is dependent on the surrounding lipid and protein components of the IMM.

59 BASIC BIOLOGICAL SCIENCES↗

Co-catabolism of arginine and succinate drives symbiotic nitrogen fixation

Biological nitrogen fixation emerging from the symbiosis between bacteria and crop plants holds promise to increase the sustainability of agriculture. One of the biggest hurdles for the engineering of nitrogen-fixing organisms is an incomplete knowledge of metabolic interactions between microbe and plant. In contrast to the previously assumed supply of only succinate, we describe here the CATCH-N cycle as a novel metabolic pathway that co-catabolizes plant provided arginine and succinate to drive the energy-demanding process of symbiotic nitrogen fixation in endosymbiotic rhizobia. Using systems biology, isotope labeling studies and transposon sequencing in conjunction with biochemical characterization, we uncovered highly redundant network components of the CATCH-N cycle including transaminases that interlink the co-catabolism of arginine and succinate. The CATCH-N cycle uses N2 as an additional sink for reductant and therefore delivers up to 25% higher yields of nitrogen than classical arginine catabolism—two alanines and three ammonium ions are secreted for each input of arginine and succinate. We argue that the CATCH-N cycle has evolved as part of a synergistic interaction to sustain bacterial metabolism in the microoxic and highly acid environment of symbiosomes. Thus, the CATCHN cycle entangles the metabolism of both partners to promote symbiosis. Our results provide a theoretical framework and metabolic blueprint for the rational design of plants and plant-associated organisms with new properties to improve nitrogen fixation.

59 BASIC BIOLOGICAL SCIENCES↗

2019 Results for Avian Monitoring at the Technical Area 36 Minie Site, Technical Area 39 Point 6, and Technical Area 16 Burn Ground at the Los Alamos National Laboratory

Los Alamos National Laboratory biologists in the Environmental Compliance and Protection Division at Los Alamos National Laboratory (LANL) initiated a multi-year program in 2013 to monitor avifauna (birds) at two open detonation sites and one open burn site on LANL property. Monitoring results from these efforts were compared among years to monitor trends. The objectives of this study were to 1) determine whether LANL operations impact bird species richness, diversity, or abundance 2) evaluate trends in species abundance by feeding guild, and 3) examine occupancy and nest success of secondary-cavity nesting birds using nestboxes. LANL biologists completed the seventh year of this effort in 2019. Three bird point count surveys were completed at each of the treatment sites at the Technical Area (TA) 36 Minie site, the TA-39 point 6, and the TA-16 burn ground between May and July 2019. A total of 853 birds representing 53 species were recorded at the three treatment sites and compared to their associated controls. Occupancy and nest success data from nestboxes at treatment sites were compared with the overall avian nestbox monitoring network. In 2019 the species richness and diversity at the treatment sites were not statistically different from their associated controls. Rarefaction and extrapolation plots for all years through 2018 suggest that over time the species diversity was statistically different between treatments and controls, although the diversity was higher at the treatment sites than the control sites. Avian abundance showed more variability but treatment and controls were trending together year to year. The mean number of granivores, insectivores, and omnivores varies from year to year, but there is not a significant trend at any of the treatment sites. The dominant feeding guild at the treatment sites continues to be insectivores. The overall results from 2019 continue to indicate that operations at the three treatment sites are not negatively affecting bird populations. This long-term monitoring will continue to monitor for any changes over time.

54 ENVIRONMENTAL SCIENCES↗

2019 Results for Avian Monitoring of Inorganic and Organic Element Concentrations in Passerine Eggs and a Nestling Collected from Technical Area 16 Burn Grounds, Technical Area 36 Minie, and Technical Area 39 Point 6 at Los Alamos National Laboratory

In 2019, non-viable avian eggs and one nestling were opportunistically collected at Los Alamos National Laboratory (LANL) near open detonation sites located at Technical Area (TA) 16 burn grounds, TA-36 Minie, and TA-39 Point 6. These samples were evaluated for inorganic elements (mostly metals), polychlorinated biphenyls (PCBs), dioxins, and furans. A total of 26 western bluebird (Sialia mexicana) and five mountain bluebird (Sialia currucoides) egg samples and one non-viable ash-throated flycatcher (Myiarchus cinerascens) nestling were collected among the three locations of interest. Concentrations of inorganic elements observed in this study were compared with the regional statistical reference level (RSRL) which is the upper-level bounds of background concentrations (mean + three standard deviations = 99% confidence interval). Several inorganic elements were not detected in avian eggs and the majority of inorganic elements detected were below the RSRL. The few elements that exceeded the RSRL were below the lowest observable adverse effect level (LOAEL), when available. One nestling collected from TA-16 contained detectable concentrations of some dioxin and furan congeners. Octachlorodibenzodioxin-1,2,3,4,6,7,8,9 concentration exceeded the RSRL, but did not exceed the calculated tetrachlorodibenzodioxin-2,3,7,8 (TCDD) toxic equivalent LOAEL. PCBs were also detected in the nestling sample and were above the RSRL but below the LOAEL. These data suggest that inorganic and organic element concentrations in eggs and nestlings are not of ecological concern. More data are needed to make a robust assessment and to evaluate trends over time.

54 ENVIRONMENTAL SCIENCES↗

Actinide Research Quarterly, First Quarter 2020: 25 Years, Commemorative Issue, 9 Articles from the Archive

This quarter we are celebrating the 25 th anniversary of Actinide Research Quarterly. The publication was established in 1995 by the former Nuclear Materials Technology Division of TA-55 at Los Alamos National Laboratory (LANL), before later becoming part of the Glenn T. Seaborg Institute and the National Security Education Center. A total of 64 issues have been produced, covering the full spectrum of topics related to actinide science and research, including contributions from esteemed scientists such as Glenn Seaborg and Siegfried Hecker. This back catalog is available electronically at www.lanl.gov/arq.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗