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Associations between Frequency of Culinary Herb Use and Gut Microbiota

While evidence suggests that culinary herbs have the potential to modulate gut microbiota, much of the current research investigating the interactions between diet and the human gut microbiome either largely excludes culinary herbs or does not assess use in standard culinary settings. As such, the primary objective of this study was to evaluate how the frequency of culinary herb use is related to microbiome diversity and the abundance of certain taxa, measured at the phylum level. In this secondary data analysis of the INCLD Health cohort, we examined survey responses assessing frequency of culinary herb use and microbiome analysis of collected stool samples. We did not observe any associations between frequency of culinary herb use and Shannon Index, a measure of alpha diversity. Regarding the abundance of certain taxa, the frequency of use of polyphenol-rich herbs and herbs with certain quantities of antibacterial compounds was positively associated with Firmicutes abundance, and negatively associated with Proteobacteria abundance. Additionally, the total number of herbs used with high frequency, defined as over three times per week, was also positively associated with Firmicutes abundance, independent of adjustments, and negatively associated with Proteobacteria abundance, after adjusting for dietary factors. Frequency of culinary herb use was not associated with Bacteroidota or Actinobacteria abundance.

59 BASIC BIOLOGICAL SCIENCES↗

Students Solving Problems for ISS and Beyond: Inspiring the Next-Generation

Students Solving Problems for ISS and Beyond: Inspiring the Next-Generation Session Title: Students Solving Problems for ISS and Beyond: Inspiring the Next-Generation Session Description: NASA High school students United with NASA to Create Hardware (HUNCH) mission is to empower and inspire students through a Project-Based Learning program where 7-12 grade students learn 21st century skills and can launch their careers through participation in the design and fabrication of real-world valued products for NASA. With six different tracks consisting of Design & Prototype, Culinary Challenge, Softgoods, Precision Machining, Software, and Video Challenge, students are given the opportunity to create solutions for the International Space Station (ISS), the Moon, and beyond. Many projects are requested by the Crew to help ease living conditions, giving students the opportunity to make an impact on the lives of Astronauts. Other projects come directly from NASA and its partners. Join our session to learn how NASA is working with teachers across the country to mentor the next generation of scientists and engineers to solve some of NASA’s greatest challenges. Learning Outcomes: 1. Describe the NASA HUNCH Program, including the program objectives, goals, and strategic partnerships for middle school and high school outreach and advocacy efforts. 2. Identify the innovative strategies NASA is using to work with middle and high school students to solve real-world problems 3. Use the knowledge gained to inspire the next generation of scientists and engineers Session Track: Advocacy & Outreach Specialized Focus Area: Women in Government and Military Learning Level: Foundational Session Format: Listen & Learn Speaker Qualifications: 1. Deboshri Sadhukhan • Current Job Title: Deputy Project Manager • Topic Experience (years of experience related to proposed topic): 1-5 years • Biography: Deboshri Sadhukhan is an engineer for NASA Glenn Research Center (GRC). She has worked on numerous projects — from International Space Station (ISS) fluid technologies, to Orion European Service Module propulsion, to planetary science missions and other game-changing technologies. Her roles have ranged from Project Manager to System Safety Lead. She currently serves as a GRC Regional Mentor for the High school students United with NASA to Create Hardware (HUNCH) program. She also serves as Deputy Project Manager for an ISS payload and Safety & Mission Assurance Lead for a Radioisotope Power Systems project. In these roles, she oversees each phase of a project from beginning to end and provides leadership to increase the reliability, maintainability and system safety of hardware and personnel throughout the system life cycle. She holds a Bachelor of Science in Electrical Engineering from The University of Akron. 2. Nancy Hall • Current Job Title: Project Manager • Topic Experience: 20+ years • Biography: Nancy Rabel Hall earned a B.S. degree in Space Sciences from Florida Institute of Technology and a M.S. degree in Mechanical Engineering from the University of Toledo. She has been at NASA Glenn for over 30 years. She has led several International Space Station experiments that studied how the behavior of fluids and fluid systems behave differently in microgravity as compared to here on Earth. She is also the High school students United with NASA to Create Hardware (HUNCH) project manager, a program that allows students to design and fabricate hardware and softgoods for NASA as well as participate in a culinary and video challenge. She enjoys talking to the public and students about the work being done at NASA as well as showing students how math and science can be fun. She is an amateur radio operator, enjoys playing golf, and reading science fiction and fantasy books.

HUNCH↗

From lab to table: Expanding gastronomic possibilities with fermentation using the edible fungus Neurospora intermedia

Fermentation is a powerful tool for enhancing flavor, improving sustainability, and expanding creative possibilities in the kitchen. However, most fermentations done in gastronomic contexts are restricted to a small set of readily available microbial species. Expanding beyond this limited biological diversity holds promise to unlock new gastronomic innovation. Here, we explore novel culinary applications of Neurospora intermedia, an edible fungus traditionally used in Java, Indonesia to produce the fermented food oncom. Our work demonstrates that N. intermedia can be readily used in the production of oncom-like meat alternatives with non-traditional substrates, as well as in the production of enzymes for starch-to-sugar conversion. As an example, we harness secreted N. intermedia starch-degrading enzymes to produce an amazake that has a distinct volatile aroma composition compared to traditional amazake produced with Aspergillus oryzae. In addition to providing texture and flavor, N. intermedia can be used to add an orange color to dishes due to the development of brightly colored spores and aerial mycelia. This property, along with its secreted enzymes, was utilized to create a dish for fine-dining restaurant Alchemist in Copenhagen, Denmark. Overall, the novel foods produced in this study were rated favorably in consumer trials, indicating a broad sensory appeal of N. intermedia across different culinary applications and cultural contexts. The protocols and approaches presented in this study represent a new addition to the chef's toolbox that hold promise to expand gastronomic possibilities with fermentation in restaurants and beyond.

59 BASIC BIOLOGICAL SCIENCES↗

Relationships between Habitual Polyphenol Consumption and Gut Microbiota in the INCLD Health Cohort

While polyphenol consumption is often associated with an increased abundance of beneficial microbes and decreased opportunistic pathogens, these relationships are not completely described for polyphenols consumed via habitual diet, including culinary herb and spice consumption. This analysis of the International Cohort on Lifestyle Determinants of Health (INCLD Health) cohort uses a dietary questionnaire and 16s microbiome data to examine relationships between habitual polyphenol consumption and gut microbiota in healthy adults (n = 96). In this exploratory analysis, microbial taxa, but not diversity measures, differed by levels of dietary polyphenol consumption. Taxa identified as exploratory biomarkers of daily polyphenol consumption (mg/day) included Lactobacillus, Bacteroides, Enterococcus, Eubacterium ventriosum group, Ruminococcus torques group, and Sutterella. Taxa identified as exploratory biomarkers of the frequency of polyphenol-weighted herb and spice use included Lachnospiraceae UCG-001, Lachnospiraceae UCG-004, Methanobrevibacter, Lachnoclostridium, and Lachnotalea. Several of the differentiating taxa carry out activities important for human health, although out of these taxa, those with previously described pro-inflammatory qualities in certain contexts displayed inverse relationships with polyphenol consumption. Our results suggest that higher quantities of habitual polyphenol consumption may support an intestinal environment where opportunistic and pro-inflammatory bacteria are represented in a lower relative abundance compared to those with less potentially virulent qualities.

60 APPLIED LIFE SCIENCES↗

Generation of a chromosome-scale genome assembly of the insect-repellent terpenoid-producing Lamiaceae species, Callicarpa americana

Plants exhibit wide chemical diversity due to the production of specialized metabolites that function as pollinator attractants, defensive compounds, and signaling molecules. Lamiaceae (mints) are known for their chemodiversity and have been cultivated for use as culinary herbs, as well as sources of insect repellents, health-promoting compounds, and fragrance. We report the chromosome-scale genome assembly of Callicarpa americana L. (American beautyberry), a species within the early-diverging Callicarpoideae clade of Lamiaceae, known for its metallic purple fruits and use as an insect repellent due to its production of terpenoids. Using long-read sequencing and Hi-C scaffolding, we generated a 506.1-Mb assembly spanning 17 pseudomolecules with N50 contig and N50 scaffold sizes of 7.5 and 29.0 Mb, respectively. In all, 32,164 genes were annotated, including 53 candidate terpene synthases and 47 putative clusters of specialized metabolite biosynthetic pathways. Our analyses revealed 3 putative whole-genome duplication events, which, together with local tandem duplications, contributed to gene family expansion of terpene synthases. Kolavenyl diphosphate is a gateway to many of the bioactive terpenoids in C. americana; experimental validation confirmed that CamTPS2 encodes kolavenyl diphosphate synthase. Syntenic analyses with Tectona grandis L. f. (teak), a member of the Tectonoideae clade of Lamiaceae known for exceptionally strong wood resistant to insects, revealed 963 collinear blocks and 21,297 C. americana syntelogs. Access to the C. americana genome provides a road map for rapid discovery of genes encoding plant-derived agrichemicals and a key resource for understanding the evolution of chemical diversity in Lamiaceae.

59 BASIC BIOLOGICAL SCIENCES↗

RNAseq analysis of Cellvibrio japonicus during starch utilization differentiates between genes encoding carbohydrate active enzymes controlled by substrate detection or growth rate

ABSTRACT Bacterial utilization of starch is increasingly of interest as the importance and contributions of animal gut microbiomes become more defined. Consequently, identifying and characterizing the bacterial enzymes responsible for the degradation, transport, and metabolism of starch will enable developments in pharmaceutical, biotechnological, and culinary industries searching for novel prebiotics, carrier molecules, and low glycemic index sweeteners. The current challenge is that bacteria proficient at starch utilization often have hundreds of carbohydrate active enzymes, and it is unclear which are essential for starch utilization using only homology-based bioinformatics or computational methods. Complementary experimental data are also needed, especially to understand the regulation of bacterial starch utilization. We have completed an RNAseq analysis of the Gram-negative bacterium Cellvibrio japonicus and found that it has sophisticated regulation that includes substrate sensing and growth rate components for genes that encode starch-degrading enzymes. Among the 22 genes predicted to encode starch-active enzymes, C. japonicus has 10 alpha-amylases, 4 alpha-glucosidases, 2 pullulnases, and 2 cyclomaltodextrin glucanotransferases, 15 of which were up-regulated during exponential growth on starch and 8 up-regulated in stationary phase. Growth analyses with an enzyme secretion deficient mutant of C. japonicus suggested that secreted amylases are essential for this bacterium to degrade starch. Our approach of coupling a physiological growth assay with transcriptomic data provides a platform to identify targets for further genetic or biochemical analysis that can be broadly applied to other starch-utilizing bacteria. IMPORTANCE Understanding the bacterial metabolism of starch is important as this polysaccharide is a ubiquitous ingredient in foods, supplements, and medicines, all of which influence gut microbiome composition and health. Our RNAseq and growth data set provides a valuable resource to those who want to better understand the regulation of starch utilization in Gram-negative bacteria. These data are also useful as they provide an example of how to approach studying a starch-utilizing bacterium that has many putative amylases by coupling transcriptomic data with growth assays to overcome the potential challenges of functional redundancy. The RNAseq data can also be used as a part of larger meta-analyses to compare how C. japonicus regulates carbohydrate active enzymes, or how this bacterium compares to gut microbiome constituents in terms of starch utilization potential.

59 BASIC BIOLOGICAL SCIENCES↗

Transmission of gravistimulus in the statocyte of the lentil root (7-IML-1)

The aim of this Biorack experiment is to determine the presentation time (minimum stimulation period to provoke a slight but significant response) of gravireaction of lentil roots cultivated in microgravity for 27 hours. The location of the statoliths in the statocytes will be analyzed in parallel in order to find the cell structure which can transmit the physical effect of the pressure of the statoliths into a biochemical factor leading to an asymmetrical production or release of the inhibitor which is responsible for the gravitropic response. Seeds of Lens culinaris will be used because of their small size. Details of the experimental procedure are given.

Perbal, Gerald↗

Consideration in selecting crops for the human-rated life support system: a Linear Programming model

A Linear Programming model has been constructed which aids in selecting appropriate crops for CELSS (Controlled Environment Life Support System) food production. A team of Controlled Environment Agriculture (CEA) faculty, staff, graduate students and invited experts representing more than a dozen disciplines, provided a wide range of expertise in developing the model and the crop production program. The model incorporates nutritional content and controlled-environment based production yields of carefully chosen crops into a framework where a crop mix can be constructed to suit the astronauts' needs. The crew's nutritional requirements can be adequately satisfied with only a few crops (assuming vitamin mineral supplements are provided) but this will not be satisfactory from a culinary standpoint. This model is flexible enough that taste and variety driven food choices can be built into the model.

NASA Center JSC↗

NASA HUNCH Hardware

What is NASA HUNCH? High School Students United with NASA to Create Hardware-HUNCH is an instructional partnership between NASA and educational institutions. This partnership benefits both NASA and students. NASA receives cost-effective hardware and soft goods, while students receive real-world hands-on experiences. The 2014-2015 was the 12th year of the HUNCH Program. NASA Glenn Research Center joined the program that already included the NASA Johnson Space Flight Center, Marshall Space Flight Center, Langley Research Center and Goddard Space Flight Center. The program included 76 schools in 24 states and NASA Glenn worked with the following five schools in the HUNCH Build to Print Hardware Program: Medina Career Center, Medina, OH; Cattaraugus Allegheny-BOCES, Olean, NY; Orleans Niagara-BOCES, Medina, NY; Apollo Career Center, Lima, OH; Romeo Engineering and Tech Center, Washington, MI. The schools built various parts of an International Space Station (ISS) middeck stowage locker and learned about manufacturing process and how best to build these components to NASA specifications. For the 2015-2016 school year the schools will be part of a larger group of schools building flight hardware consisting of 20 ISS middeck stowage lockers for the ISS Program. The HUNCH Program consists of: Build to Print Hardware; Build to Print Soft Goods; Design and Prototyping; Culinary Challenge; Implementation: Web Page and Video Production.

manufacturing↗

NASA’s Continued Partnerships with High School Students during a Global Pandemic

NASA HUNCH’s mission is to empower students through Project-Based Learning where they learn skills and can launch their careers through participation in the design and fabrication of real-world valued products. Our goal is to mentor the next generation of students to solve NASA’s greatest challenges even during a global pandemic

HUNCH↗