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Full-Text Articles in Integrative Biology

Coralai: Emergent Ecosystems Of Neural Cellular Automata, Aidan A. Barbieux, Aidan A. Barbieux Mar 2024

Coralai: Emergent Ecosystems Of Neural Cellular Automata, Aidan A. Barbieux, Aidan A. Barbieux

Master's Theses

Artificial intelligence has traditionally been approached through centralized architectures and optimization of specific metrics on large datasets. However, the frontiers of fields spanning cognitive science, biology, physics, and computer science suggest that intelligence is better understood as a multi-scale, decentralized, emergent phenomenon. As such, scaling up approaches that mirror the natural world may be one of the next big advances in AI. This thesis presents Coralai, a framework for efficiently simulating the emergence of diverse artificial life ecosystems integrated with modular physics. The key innovations of Coralai include: 1) Hosting diverse Neural Cellular Automata organisms in the same simulation that …


Steminism: Analyzing Factors That Improve Retention Of Women In Stem, Kira Carter, Jane Kelley, Jason Vasser-Elong, Rc Patterson Feb 2024

Steminism: Analyzing Factors That Improve Retention Of Women In Stem, Kira Carter, Jane Kelley, Jason Vasser-Elong, Rc Patterson

Dissertations

Our co-authored research ‘Steminism: Analyzing Factors That Improve Retention for Women as STEM Majors’ analyzed factors that contributed to the retention of women in science, technology, engineering, and mathematics (STEM) programs at Missouri University of Science & Technology (Missouri S&T). Women make up half of the US population, and while careers in (STEM) are an integral part of the US economy, women are underrepresented in these career fields. The purpose of our dissertation is to address the underrepresentation of women in STEM majors. Our methodology included homogeneous sampling to collect qualitative data. More specifically, we consulted with academic advisors and …


Parental Environnement Influences Offspring Gene Expression In The Olympia Oyster, Ostrea Lurida., Mark S. Yeats Jan 2024

Parental Environnement Influences Offspring Gene Expression In The Olympia Oyster, Ostrea Lurida., Mark S. Yeats

LSU Master's Theses

The Olympia Oyster is the only native oyster species on the Pacific coast of North America. Seasonality in water temperatures across this threatened species’ broad and fragmented range has been linked to its development, reproductive timing, reproductive activity, growth, and survival rates. Recent evidence suggests parental environments can have carry-over effects on larval growth and respiration rates in response to temperature. In this study, we utilized transcriptomic methods to identify the underlying mechanisms of this cross-generation plasticity. Sequence libraries of adult ctenidia tissue and pooled larval replicates were extracted and spawned from hatchery raised oysters of the same parentage immediately …


The Cetacean Sanctuary: A Sea Of Unknowns, Jason N. Bruck Jan 2024

The Cetacean Sanctuary: A Sea Of Unknowns, Jason N. Bruck

Faculty Publications

Housing cetaceans in netted sea pens is not new and is common for many accredited managed- care facilities. Hence, the distinction between sanctuary and sea pen is more about the philosophies of those who run these sanctuary facilities, the effects of these philosophies on the animals’ welfare, and how proponents of these sanctuaries fund the care of these animals. Here, I consider what plans exist for cetacean sanctuaries and discuss the caveats and challenges associated with this form of activist-managed captivity. One goal for stakeholders should be to disregard the emotional connotations of the word “sanctuary” …


Development And Evaluation Of A Mitochondrial Dna Marker For The Detection Of Rattus Rattus And Rattus Norvegicus Fecal Contamination, A'Aden Redhead Jan 2024

Development And Evaluation Of A Mitochondrial Dna Marker For The Detection Of Rattus Rattus And Rattus Norvegicus Fecal Contamination, A'Aden Redhead

Graduate Research Showcase

No abstract provided.


Environmental Biology Masters Capstone, Antonio Gonzalez-Pita Jan 2024

Environmental Biology Masters Capstone, Antonio Gonzalez-Pita

Regis University Student Publications (comprehensive collection)

Human wildlife interactions (HWI) pose a complex challenge for wildlife managers. Human encroachment into wildlife habitat and the growing number of outdoor recreationists are increasing the frequency of contact and conflict, especially in regions such as the Front Range of Colorado. Geographic information systems (GIS), which use a combination of remote sensing and environmental survey data, allow for predictive spatial analyses of where human wildlife interactions are likely to occur. I used publicly reported observations of moose to create spatial predictive maps in a species distribution model framework. Slope and elevation were shown to be the strongest predictors of HWI, …


Managing Stress: A Study Of Stress Response Mechanisms In Mycobacteria, Augusto C. Hunt Serracin Jan 2024

Managing Stress: A Study Of Stress Response Mechanisms In Mycobacteria, Augusto C. Hunt Serracin

Biology Dissertations - Archive

Mycobacteria encompass many pathogenic species known to cause severe disease in humans. A well-known example is Mycobacterium tuberculosis (Mtb), the causative agent of the lung disease tuberculosis, which kills millions of humans worldwide yearly. Pathogenic mycobacteria like Mtb are challenging to treat because of their innate ability to adapt to environmental stress. Their unique cell physiology and conserved stress responses allow them to combat biological insults, regulate growth, and regulate genes involved in stress; all these responses increase tolerance to antibiotics. The current therapies to treat mycobacterial infections are lengthy and, at times, unsuccessful, partly due to antibiotic tolerance. A …


The Morphological Evolution Of The Inner Ear In Lacertoidea (Squamata), Cristian Hernandez Jan 2024

The Morphological Evolution Of The Inner Ear In Lacertoidea (Squamata), Cristian Hernandez

Biology Dissertations - Archive

The popularization of high-resolution computed tomography(HRCT) has produced a revolution in the field of comparative anatomy. This technology allows us to reach a level of morphological detail in our observations that has never been possible previously. Now we can perform digital dissections isolating structures of interest such as individual bones, soft tissue organs, and even generate 3D models of body cavities (endocasts) such as the braincase and inner ear. Encompassed with the progress of the HRCT, 3D geometric morphometrics have emerged as a common tool to address morphological questions in a statistical context that outperforms the traditional 2D geometric morphometrics. …


Corals In Crisis: A Transcriptomic Investigation Of Stony Coral Tissue Loss Disease, Kelsey M. Beavers Jan 2024

Corals In Crisis: A Transcriptomic Investigation Of Stony Coral Tissue Loss Disease, Kelsey M. Beavers

Biology Dissertations - Archive

Coral reefs face existential threats due to climate change and human activities. The combined effects of ocean warming, overfishing, pollution, and habitat destruction have fundamentally altered the functionality of coral reefs and are driving a drastic increase in the prevalence and severity of coral disease outbreaks. Stony coral tissue loss disease (SCTLD) has emerged one of the most devastating disturbances to Caribbean reef ecosystems on record. Despite notable progress, the cellular mechanisms driving SCTLD pathogenesis remain poorly understood. To address this issue, my dissertation investigates the gene expression, histopathology, immune protein activity, and symbiont community dynamics involved in SCTLD progression …


An Updated Outline For Clearing And Staining Of Preserved Specimens, Sadie E. Smith Jan 2024

An Updated Outline For Clearing And Staining Of Preserved Specimens, Sadie E. Smith

Honors Theses and Capstones

This protocol outlines a enhanced clearing and staining procedure for persevered specimens, focusing on the visualization of cartilage and bone in small taxa. With origins from O. Shultze's diaphonization methodology in 1897, the process of clearing and staining improves the clarity and contrast of intricate tissues. It is essential for studies in developmental biology, histology and anatomy. This protocol covers the key steps of the process while detailing the materials, solutions and estimated timeline for each step pertaining to specific taxa. The aim of the protocol is to standardize and provide guidance for procedures in the Biological Sciences department at …


The Biobehavioral Effects Of Embryonic Exposure To Neural Inflammation And Oxidative Stress In Zebrafish, Dalton J. Anderson Jan 2024

The Biobehavioral Effects Of Embryonic Exposure To Neural Inflammation And Oxidative Stress In Zebrafish, Dalton J. Anderson

Honors College Theses

The purpose of this research is to improve understanding of the neurodevelopmental effects of embryonic exposure to elevated inflammation and oxidative stress induced by the antipyretic drug acetaminophen (APAP). Our study was the first to examine the interactive effects of APAP and inflammation in zebrafish embryos and how the treatments affect brain development and larval behavior. Experimental groups of zebrafish larvae were exposed to lipopolysaccharide (LPS) to induce inflammation, APAP, or LPS + APAP and larval behavior was analyzed using Ethovision automated behavioral tracking software. We also measured changes in whole-brain Glycogen Synthase Kinase 3 Beta (GSK3B) and GSK3B phosphorylation, …


Female Titin Mutant Mice Outperform Young And Old Males, Reese Ger Jan 2024

Female Titin Mutant Mice Outperform Young And Old Males, Reese Ger

Scripps Senior Theses

Exercise performance is widely believed to generally be maximal among young males. However, little research has been conducted to investigate the impact of sex or age on muscle performance. This study investigated the effects of sex and age on forelimb and all limb grip strength performance in wildtype and titin mutant mice. Titin is the largest known protein, with known involvement in muscle function. The mice in this study were Ttn∆112-158 with a 75% deletion to the PEVK region of titin. Based on limited existing research, we hypothesized that wildtype mice would outperform all Ttn∆112-158 mice, males would outperform females …


Microglia Regulate Light Evoked Retinal Responses In The Zebrafish Retina., Brittney Moore Jan 2024

Microglia Regulate Light Evoked Retinal Responses In The Zebrafish Retina., Brittney Moore

Williams Honors College, Honors Research Projects

The primary goal of the project is to investigate the role of the interferon regulatory factor 9 (irf8) gene in the retinal function of zebrafish. My research aims to examine and compare the retinal responses of two subject groups: wildtype zebrafish and zebrafish with the irf8 gene knocked out. This project seeks to determine if irf8 knockout zebrafish lose visual system functionality before experiencing changes in retinal morphology. The primary technique to answer this research question is the electroretinogram, a commonly used method to examine retinal function. The final outcome of this project will be a scientific paper describing the …


The Effects Of Ethanol Exposure On Glucose And Lactate Levels In Crayfish, Mackenzie Kirwan Jan 2024

The Effects Of Ethanol Exposure On Glucose And Lactate Levels In Crayfish, Mackenzie Kirwan

Williams Honors College, Honors Research Projects

The goal of the project is to investigate and determine the effects of sunlight exposure and hormone levels. The circadian rhyth, which is heavily influenced by sunlight, not only affects the amount of sleep someone achieves, but is also related to behavior and mood, and heavily relies on the body’s hormones for proper regulation. The hormones that are prevelant to the circadian rhythm, and which will be studied in this experiment are cortisol, serotonin and melatonin. In this experiment, these three hormones will be extracted prior to alterations of light exposure. Animal test subjects will then undergo treatment of light …


Snake Translocation Policies And Practices Within The United States, Robin E. Bedard, Megan Rottenborn, Emily Taylor Jan 2024

Snake Translocation Policies And Practices Within The United States, Robin E. Bedard, Megan Rottenborn, Emily Taylor

Human–Wildlife Interactions

Human–wildlife conflict with nuisance snakes (Serpentes) is rapidly increasing with human population growth and development rates. Translocation of nuisance snakes has become a widespread practice, aided by social media pages connecting people to volunteer and for-profit snake translocators. However, translocators often struggle to find information and resources from wildlife agencies about the policies required, and some may unintentionally use translocation procedures that disregard the health and survival of the snakes. The goals of this study were to (1) obtain data on policies, permitting, and training required for translocating nuisance snakes in each U.S. state that has snakes and (2) compare …


Restoration For The Small And Slimy: How Pond-Breeding Amphibians Utilize Natural, Restored, And Created Wetlands, Miranda Florent Jan 2024

Restoration For The Small And Slimy: How Pond-Breeding Amphibians Utilize Natural, Restored, And Created Wetlands, Miranda Florent

Williams Honors College, Honors Research Projects

This research project for the Williams Honors College sought to examine how local pond-breeding amphibians utilized natural, restored, and created wetlands. Presence/absence data was gathered from field surveys, historical records, and iNaturalist data for the six study sites. It was predicted that created and restored wetlands may support less sensitive species with lower coefficient of conservatism scores, based on past surveys of a similar nature. No statistically significant results were found in this survey, but that could be due to low sample sizes and limited data sets, among other considerations. Therefore, more in-depth studies should still be undertaken to examine …


The Influence Of Genes On The Migratory Timing Of Great Reed Warblers, Emily Fackler Jan 2024

The Influence Of Genes On The Migratory Timing Of Great Reed Warblers, Emily Fackler

Honors Undergraduate Theses

Many organisms migrate; however, there is a lack of knowledge on the mechanisms that drive migration timing, including which genes play a role. The great reed warbler (Acrocephalus arundinaceus), a migratory bird that breeds in Eurasia and overwinters in sub-Saharan Africa, is an excellent model to study the genetics of migration. Each season, some individuals migrate early, whereas others migrate later in the season. In other words, they exhibit differences in their migratory chronotypes. Past research has identified three candidate genes (TOP1, PEAK1, and CPNE4) that influence the migratory chronotypes of another avian species, …


Interdisciplinary Learning: How Different Ways Of Thinking Can Fuel Passion And Expand Understanding., Allison Amy Peschek Jan 2024

Interdisciplinary Learning: How Different Ways Of Thinking Can Fuel Passion And Expand Understanding., Allison Amy Peschek

Undergraduate Theses, Professional Papers, and Capstone Artifacts

This Capstone project was created to show the profound experience resulting from an extended range of interdisciplinary learning, and how different methods, when paired together, can enhance a learning experience. This was a personal project of exploration into those different angles, and primarily focused on avenues of logical and creating learning, respectively. These methods have been implemented by this author often in the past, but this capstone will explore this concept further, as there is a lot of potential in combining these different methods and biases in learning, to both further passion and understanding as a learner, but to also …


Eliciting The Effect Free Roaming Cats Have On Native Hawaiian Wildlife Using Stable Isotope Analysis, Erin Salano Jan 2024

Eliciting The Effect Free Roaming Cats Have On Native Hawaiian Wildlife Using Stable Isotope Analysis, Erin Salano

Theses and Dissertations--Biology

This study measured the concentration of the stable isotopes Carbon and Nitrogen within the tissues of free-roaming and indoor cats on the Hawai’ian islands. This study aimed to determine whether cats are consuming more human provisioned or wild food sources and whether their diet is having a negative impact on endemic species; with a focus on assessing impacts on conserved bird populations. We hypothesized that human provisioned food sources have higher levels of carbon due to the amount of corn fillers within them, while wild food sources are more Nitrogen rich. We found that indoor and feral cats sampled from …


Incorporating Sex-Diverse And Gender-Inclusive Perspectives In Higher Education Biology Courses, Breanne N. Harris, A Kelsey Lewis, Sam L. Sharpe, Teri J. Orr, Christopher T. Martine, Chloe C. Josefson Jan 2024

Incorporating Sex-Diverse And Gender-Inclusive Perspectives In Higher Education Biology Courses, Breanne N. Harris, A Kelsey Lewis, Sam L. Sharpe, Teri J. Orr, Christopher T. Martine, Chloe C. Josefson

Faculty Journal Articles

Inclusive teaching is teaching in a way that reaches all students in the classroom; this is beneficial for everyone, particularly for those with minoritized identities. Instructors play a critical role in scaffolding how students are exposed to and learn science content in the classroom. In this manuscript, we discuss how biology instructors can make their classrooms more inclusive with regard to sex and gender diversity content. Many topics in biology are based on androcentric, heteronormative, and oppressive framing, even though those lenses are more reflective of our own history and culture than they are of the diversity we see in …


How Environmental Variation Affects Bumblebee Cognition And The Gut Microbiome, Parker Madison Campbell Jan 2024

How Environmental Variation Affects Bumblebee Cognition And The Gut Microbiome, Parker Madison Campbell

Graduate Theses/Dissertations

This master's thesis examines the influence of environmental variables on cognitive processes in Bombus impatiens, focusing on the effects of ingested microbial communities on gut microbiota and their subsequent impact on associative color learning. The study implemented controlled feeding trials with isolated microbe species (Candida bombi, Metschnikowia reukaufii, Lactobacillus micheneri and a control with no microbes) to analyze shifts in gut microbial composition in foraging bees. Results indicated minor alterations in the microbial profiles of bees, yet no statistically significant correlation was found between these modifications and variations in learning or memory performance. Furthermore, the study assessed the impact of …


Evaluation Of Alternative Methods For Dna Recovery Of Compromised Skeletal Remains, Ciara N. Rhodes Jan 2024

Evaluation Of Alternative Methods For Dna Recovery Of Compromised Skeletal Remains, Ciara N. Rhodes

Theses and Dissertations

Skeletal remains frequently serve as the sole source of genetic material available for developing a profile in cases concerning unidentified human remains or mass casualties. Nevertheless, as unidentified skeletal remains age and are exposed to uncontrolled environmental conditions, the success rates of conventional methods of bone analysis diminish significantly. The physical and mechanical processing of skeletal remains into fine powder for DNA isolation and downstream analysis can substantially impact the quality and quantity of recoverable genetic information. This concern may be exacerbated when dealing with degraded forensic skeletal remains. Few studies have explored whether pulverization of bone samples contributes to …


Characterizing Leaf Endophyte Composition In The Dune Grass Ammophila Breviligulata, Nabaa Majeed, Sarah Hobbs, Sarah Emery, Connor Morozumi, Natalie Christian Jan 2024

Characterizing Leaf Endophyte Composition In The Dune Grass Ammophila Breviligulata, Nabaa Majeed, Sarah Hobbs, Sarah Emery, Connor Morozumi, Natalie Christian

Undergraduate Research Events

Microorganisms living within plants play crucial roles in their health and function, yet these diverse communities that make up the plant microbiome remain poorly understood. The dune grass Ammophila breviligulata. is an ecosystem engineer which helps to build and stabilize dune ecosystems effectively acting as a buffer against storms to coastal communities. It is currently unknown whether the Ammophila microbiome plays a role in this plant species’ ability to transform dune ecosystems.. In this study, we assessed fungal diversity of endophytes—microbes living within plant tissues— in Ammophila leaves collected from Leelanau State Park, MI along the eastern shores of Lake …


Effect Of Rare Earth Element Mixtures On Daphnia Magna, Celine Do Jan 2024

Effect Of Rare Earth Element Mixtures On Daphnia Magna, Celine Do

Theses and Dissertations (Comprehensive)

Rare earth elements (REEs) are increasingly being used in modern technologies and as such, there is an increasing concern for the potential environmental risk associated with anthropogenic contamination. In addition to there being very little data available for individual REEs, there is also a lack of knowledge concerning mixtures. The objective of my study was to investigate the chronic effects of lanthanum (La) and yttrium (Y) as well as the acute effects of neodymium (Nd), praseodymium (Pr) and Y as a single metal and as a mixture to Daphnia magna. Standard 21-d chronic tests and 48-h single metal acute …


Comparing Ionoregulation And Modes Of Nitrogen Excretion Across The Life Cycle Of Parasitic And Non-Parasitic Lamprey Species, Shannon Davidson Jan 2024

Comparing Ionoregulation And Modes Of Nitrogen Excretion Across The Life Cycle Of Parasitic And Non-Parasitic Lamprey Species, Shannon Davidson

Theses and Dissertations (Comprehensive)

Abstract

Ionoregulation and nitrogenous waste (N-waste) metabolism in fishes has been extensively documented in teleosts and elasmobranchs, but less so in lampreys (Petromizontiformes). All lamprey species undergo metamorphosis which drastically changes their physiology and modes of feeding. However, differences in ionoregulatory mechanisms due to anadromous versus non anadromous life history strategies emerge following metamorphosis in different lamprey species. Furthermore, some species, such as anadromous juvenile sea lamprey (Petromyzon marinus), are parasitic and migrate to seawater (SW) to ingest blood protein from host fishes, while juveniles of freshwater (FW), non-parasitic species, such as the American brook lamprey …


Investigating The Role Of The Mammalian Dsrnai Pathway In Preventing Infection By Respiratory Syncytial Virus And Human Coronavirus, Iliana Portelli Jan 2024

Investigating The Role Of The Mammalian Dsrnai Pathway In Preventing Infection By Respiratory Syncytial Virus And Human Coronavirus, Iliana Portelli

Theses and Dissertations (Comprehensive)

dsRNA interference (dsRNAi) is an antiviral innate immune response that uses long dsRNA (>40bp) to inhibit gene expression through the RNA interference (RNAi) pathway. dsRNAi is an important antiviral defense mechanism in plants, invertebrates, and fungi. However, its use as an antiviral mechanism in vertebrates is not well understood. Evidence proving the significance of dsRNAi in vertebrate cells does exist, but there is still a lack of research in this area, particularly with the use of long dsRNA. Respiratory Syncytial Virus (RSV) and human coronaviruses (HCoVs) are two single stranded RNA (ssRNA) viruses that infect airway cells. This study …


Elucidating Mechanisms Of Enhanced Dsrna-Nanophytoglycogen Innate Immune Responses In Healthy Human Cells, Nicholas Jadaa Jan 2024

Elucidating Mechanisms Of Enhanced Dsrna-Nanophytoglycogen Innate Immune Responses In Healthy Human Cells, Nicholas Jadaa

Theses and Dissertations (Comprehensive)

Immunostimulatory nucleic acids, such as long double-stranded RNA (ds)RNA, can stimulate innate immune responses in a non-sequence specific manner. These molecules are recognized by pattern-recognition receptors in the cytoplasm, endosome, and on the cell surface. Activation leads to the production of mediators of innate immune pathways, including type I interferon and proinflammatory cytokines. Although these receptors are present in all cells, their role in non-immune cells is often overlooked due to their lower level of responsiveness. Nanocarriers can enhance these nucleic acid-mediated responses, allowing for exploration of innate immune pathways in non-immune cells. Nanoparticles as carriers of nucleic acids can …


The Effects Of Warmer Temperatures On The Physiology Of Invasive Sea Lamprey And Imperiled Populations Of Lake Sturgeon In The Laurentian Great Lakes, Milica Koledin Jan 2024

The Effects Of Warmer Temperatures On The Physiology Of Invasive Sea Lamprey And Imperiled Populations Of Lake Sturgeon In The Laurentian Great Lakes, Milica Koledin

Theses and Dissertations (Comprehensive)

Invasive sea lamprey (Petromyzon marinus) contributed to massive declines in fisheries in the Laurentian Great Lakes due to their parasitism of large-bodied salmonid fishes in the 20th century. Chemical control measures, which involve the application of 3-trifluoromethyl-4-nitrophenol (TFM) into rivers containing larval sea lamprey, were successfully implemented approximately 60 years ago to control populations. Recently, however, it was discovered that TFM tolerance increases in sea lamprey with a rise in temperature for reasons which are incompletely understood. Nor is it understood how sea lamprey will respond to warmer temperatures due to climate change, which could affect their …


Impacts Of Environmental Stressors On Mummichog (Fundulus Heteroclitus) Growth And The Gh-Igf1 Pathway Using A Laboratory Bioassay, Olena Kuntyj Jan 2024

Impacts Of Environmental Stressors On Mummichog (Fundulus Heteroclitus) Growth And The Gh-Igf1 Pathway Using A Laboratory Bioassay, Olena Kuntyj

Theses and Dissertations (Comprehensive)

Using a flow-through laboratory bioassay, experiments investigated the effects of limited feeding, increased temperature, ammonium chloride (NH4Cl), and 17α-ethinylestradiol (EE2) on juvenile mummichog (Fundulus heteroclitus) growth. Weight measurements were taken every three days as the apical growth endpoint. Gene expression was examined by qPCR for genes in the GH (growth hormone)-IGF1 (insulin-like growth factor 1) pathway, including gh, igf1, and their receptors (ghra, ghrb, igf1ra, igf1rb). In Experiment 1, juvenile mummichog were exposed to excess or limited feed for 21 days. The limited-fed mummichog weighed less, were …


High-Quality Dietary Protein (Essential Amino Acids Matched To Reproductive Needs) Partially Breaks The Lifespan And Reproduction Trade-Off In Lubber Grasshoppers, Emma Kordek Jan 2024

High-Quality Dietary Protein (Essential Amino Acids Matched To Reproductive Needs) Partially Breaks The Lifespan And Reproduction Trade-Off In Lubber Grasshoppers, Emma Kordek

UNF Graduate Theses and Dissertations

Finding interventions to break the trade-off between reproduction and lifespan can provide insight into physiological limitations of animals. Effects of dietary protein quality on the trade-off are currently unclear, but clarity could lead to better designed diets that match animal needs. Dietary amino acid blends matching yolk proteins support reproduction and extend lifespan in fruit flies. To test if this is conserved across species, we matched dietary amino acids to vitellogenin to test reproduction and lifespan in adult females of the lubber grasshopper. Specifically, we compared varying degrees of protein quality by manipulating dietary essential amino acids. We identified a …