Research utilizing human and non-human animal analogues has provided valuable information to forensic anthropology. This includes data on taphonomic processes that have assisted in the refinement of forensic archaeological techniques that are crucial to locating clandestine graves, recovery of decedents, and scene documentation. Additionally, research on the effects of various trauma mechanisms on skeletal remains has allowed for increased involvement of forensic anthropologists in trauma-specific casework. Animal models have long been used to replicate humans, dating back to early anatomical and biomedical research. However, despite their widespread use, recent studies have called into question the legitimacy of research involving animal models. While the number of outdoor taphonomic facilities using human donors continues to expand, access to such resources remains limited. As a result, a substantial portion of current forensic anthropological data is still derived from studies using non-human analogues. These models often serve as practical alternatives where human donors are unavailable, though their limitations must be acknowledged. This presentation highlights the contributions of non-human animal proxies in forensic anthropological research and discusses when their use may be appropriate. Guidelines are proposed for evaluating proxy suitability, including the overarching goals of the project, what research has previously been conducted, the availability of different species and locations for research, as well as the ethical considerations of the research. Thus, before discounting non-human animal proxies entirely, their distinct advantages and disadvantages should be evaluated for each research goal during the project design process to determine the applicability of proxy use in forensic anthropological research. 

The regulation of the cell cycle is essential for maintaining cell health and preventing cellular diseases like cancer. The budding yeast Saccharomyces cerevisiae is widely used as a eukaryotic model to investigate these mechanisms because of its rapid cell cycle and ease of genetic manipulation. Elm1, a protein kinase, has been shown to regulate the septin cytoskeleton, mitotic exit, and cell polarity; however, the process behind these connections remains a mystery. A recent proteomic screen identified the polarity protein Boi1 as a potential Elm1 substrate. In this study we investigated Boi1 and paralog Boi2 for their effects on the Mitotic Exit Network (MEN). Fluorescence microscopy revealed that Boi1 and Boi2 localize to septin structures with timing like the MEN phosphatase, Cdc14. The absence of Boi1 leads to premature Cdc14 recruitment. The removal of Boi2 produced more subtle effects on the cell cycle timing, as shown by the microtubules, with no statistically significant differences in the microtubule dynamics. Cells lacking Boi2 had an increase in transitional double-ring septins, while Cdc14 was localized earlier than normal. Together, these results highlight distinct but similar roles for Boi1 and Boi2 in coordinating septin architecture with MEN signaling. Boi1 appears to play a more dominant role in the regulation of the cell cycle, while Boi2 refines the coordination of mitotic exit events. This research helps provide new information about how polarity proteins work together with signaling networks to ensure proper cell division and offers a baseline for understanding similar regulatory mechanisms in humans.
 

Glioblastoma(GBM) is one of the most aggressive brain cancers, causing around 200,000 deaths annually in the United States. The phosphoacetylglucosamine mutase(PGM3) has been recently identified as a potential therapeutic target for GBM. The goal of this study is identifying a monoclonal antibody that binds effectively to PGM3 using computational modeling and molecular docking techniques, leading to the effective treatment and detection of GBM . The amino acid sequence for PGM3 and antibodies was obtained from UniProt and modeled by the AI system AlphaFold3, effectively predicting its 3D structure. Multiple antibodies then underwent docking simulation with PGM3 using the HDOCK machine learning model. PLIP was then used to analyze the interactions such as hydrogen bonds and π-π stacking in the docking, while Prodigy (Machine learning model) estimated binding affinity in ΔG (kcal/mol). Using P2Rank(Machine learning model) and ScanNet (AI deep learning model), the binding site of PGM3 was identified. Plugging the previous data into an antibody selection criteria, antibody 6KDH was identified as the ideal candidate. To further validate the effectiveness of 6KDH, molecular dynamic simulation was performed using GROMAC software, which confirms that 6KDH has a stable bond with the PGM3 protein making it the most ideal antibody out of the antibodies tested. These results show that antibody 6KDH may serve as a framework for future computational as well as laboratory validation and could be a starting point in the treatment and detection of GBM. 
 

Crucial to our understanding of speciation is how populations diverge and accumulate differences. Walnut-infesting Rhagoletis speciated allopatrically with geographic isolation as the primary barrier to reproduction between species. When this barrier is experimentally removed, sexual isolation is maintained, suggesting behavioral mechanisms underlie mating decisions between closely related species. We performed mate choice assays by pairing a female R. juglandis with a conspecific and a heterospecific male. We found strong assortative mating (91% of females mating within species) which is not surprising given ~0.5 million years of divergence and morphological differences, notably in body size.

To further understand how sexual isolation develops within species, we conducted behavioral trials between two geographically separated (~150 km) populations of R. juglandis. We also measured body size to evaluate morphological variation. We found that individuals from one population were significantly larger. However, we found no evidence of assortative mating; only 42% of copulations were between flies of the same population. These results suggest that morphological divergence between populations may precede divergent mate choice preferences. While species of Rhagoletis exhibit strong sexual isolation in experimental settings, geographically separated populations of R. juglandis do not, despite evolving morphological differences. Divergence in courtship behavior may additionally clarify patterns of sexual isolation. Understanding the full scope of variation within and between Rhagoletis species can clarify how mating decisions contribute to early stages of speciation.

The Nancy Member of the Borden Formation of central Kentucky contains an assemblage of Osagean trilobites which includes new occurrences of several species, extends the range of an existing genus, and contains an occurrence of one new species. Two species from the assemblage have been previously recorded from the Borden Formation, Phillibole conkini and Australosutura lodiensis, but only in its lowermost member: the New Providence Shale. Breviphillipsia semiteretis has been recorded from several occurrences throughout the United States, but never in the Borden Formation. Namuropyge (Cuyapyge) brooksi has been recorded from earlier (Kinderhookian) strata in the United States, but the occurrence of this genus in the younger (Osagean) strata of the Borden Formation marks an extension to the chronostratigraphic range of this genus in North America. The new species Australosutura sp. nov. is distinguished from all other known Australosutura species by the unique array of spines along the margins of its cephalon and pygidium, as well as by its glabella shape and overall tuberculation pattern. The occurrence of Australosutura lodiensis and Australosutura sp. nov. in the same assemblage is significant because it marks the first known cooccurrence of multiple species of this genus.

Fluorescent dye tracing is essential for delineating groundwater flow in karst aquifers, where subsurface connectivity governs water quality, protection, and contaminant transport. Charcoal receptors are standard, yet construction materials may bias dye recovery. Despite widespread use, few field studies quantify material effects. Controlled tests by Crawford Hydrology Lab (CHL) and the Kentucky Geological Survey (KGS) suggested little difference between fiberglass mesh bags and filter fiber “milk socks,” but field deployments introduce variables that could alter performance. This study presents the largest field-based comparison of these two receptor types.
To evaluate these differences, CHL, in partnership with U.S. Geological Survey (USGS), conducted a series of multi-dye tracer tests using paired mesh and milk-sock receptors. Tracers included Tinopal, Fluorescein, Eosine, Rhodamine WT, and Sulphorhodamine B. Of 1,043 paired samples, 312 yielded measurable dye; the remainder were pairs of non-detects. Among the 312 positive pairs, 72% of mesh receptors recorded concentrations more than 10% higher than their paired milk socks, and 35% were more than 50% higher. Several mesh samples required dilution due to high concentrations, whereas paired milk socks did not. Milk socks frequently contained fine charcoal dust, elevating background signal and complicating peak interpretation.
These results indicate that mesh bags more consistently yield higher dye recoveries than milk socks. While milk socks provide logistical and cost advantages, their use increases the risk of underestimating dye presence. Understanding material limitations will improve receptor selection and interpretation in karst dye tracing and strengthen decision-making for water resource protection.

Persulfates are compounds capable of oxidizing a wide range of organic materials. They can be used in soil and groundwater remediation to break down organic contaminants. More than 50% of the U.S. population relies on groundwater for drinking water, which is also one of the most important sources of water for irrigation. Unfortunately, groundwater is highly susceptible to pollution. Fluorescent dyes are organic compounds used to detect and quantify the presence of target molecules. The use of these dyes can help trace the direction of groundwater flow and determine travel times. The goal of this research is to understand how persulfate oxidizing agents interact with fluorescent dyes. Samples collected in the field and stored under subsurface-like conditions were analyzed using a field fluorometer. Insights into the behaviors of contaminant oxidation by persulfates, as well as the fluorescence dye, will be presented.
 

National research statistics reported that the six-year certificate and degree completion rates of students differ by race and ethnicity. For African American students, about 29% completed their degree, and 45% were no longer enrolled in college at the end of the six-year timeframe. The corresponding average values for non-minority students are 43% and 31%, respectively. Although the literature explores several explanations for this phenomenon, few studies have examined this topic from the perspective of African American high school students from Kentucky who plan to attend college and major in a Science, Technology, Engineering, and Mathematics (STEM) field or program. The purpose of this study is to listen to the voices of African American high school students from Louisville, Kentucky and identify the extent to which their level of motivation, self-efficacy, mentorship, and family support are aligned with a success mindset when they face their freshman year, and what obstacles the participants think they may face as they transition from high school to college in a STEM major. It is expected that the researchers' findings will result in actionable recommendations to minimize attrition and promote collaboration among communities, higher education personnel, and policymakers to implement effective retention strategies.

  Lipopolysaccharides (LPS), also known as endotoxin, is a component of Gram-negative bacterial cell walls that are known to activate immune responses but also directly affect ion channels. LPS, from Serratia marcescens, results in a transient hyperpolarization of the membrane potential, followed by a subsequent depolarization in larval Drosophila muscle. The mechanisms of action appears to be that LPS transiently activates two-pore-domain K+ (K2P) channels, followed by action on the Na+ leak channels (NALCN) channels. Substitution of the ionic composition and concentrations of ions in the bathing media, as well as genetic overexpression of dORK1 (i.e., a Drosophila K2P subtype), helped to substantiate the proposed mechanisms. However, the effects on the overall permeability of the ions in varied conditions have yet to be estimated. With the use of the Goldman-Hodgkin-Katz (GHK) equation, the known concentrations of ions in the bathing media, and empirical data of the membrane potential, the focus is now to estimate the permeability of the ions with computational simulations. In this study, through computational simulations, the GHK equation provided an accurate framework for estimating ionic permeabilities during LPS-induced changes in membrane potential, highlighting the use of the GHK equation in modeling complex ion channel dynamics.
 

     Where and with whom do students of color (SOC) find a sense of belonging in college? SOCs currently make up 33% of undergraduate students and projections suggest that they will reach as much as 57% by 2050. Compared to their White classmates, SOCs have shown lower rates of persistence towards completing a degree in higher education. Sense of belongingness is a key predictor of students’ persistence towards graduation. At Centre College, as a PWI, we are focused on understanding where and with whom SOC find a sense of belonging. In the present research, we conducted 21 semi-structured interviews with students of color (10 Men, 10 Women, 1 Non-binary) at Centre College. Participants discussed spaces where they do and do not belong, their transition to college and support from family, and the role of language in their experiences. Qualitative coding revealed that counterspaces and shared identity with other students contributed to a sense of belonging. Our findings also point to a key role of white professors and professors of color contributing to a sense of belonging both in and outside of the classroom. Students shared being misunderstood and feeling ‘othered’ because of cultural and language differences. Some SOCs also attributed difficulties to their perceived lack of academic preparation. In this presentation, we will discuss initial findings and identify contributors towards feelings of belonging and not belonging in college.

Evaluating how much pain one is in given pain facial expressions (PFE) can be difficult. Numerous scales exist to assist health care professionals in documenting pain experienced by patients. Despite strong evidence that pain is often displayed with a narrow set of facial muscle activation, even trained human coders have difficulty distinguishing between genuine and posed expressions of pain. We compared an existing data set created by humans who observed a dynamic set of pain expressions and coded them for facial cue activation to coding data newly generated by the OpenFace software platform. OpenFace allows users to estimate facial action coding unit activation in videos, recording the presence and proportional strength of cues. We evaluated the extent to which outcomes from human coders coding and from automated platforms were similar, especially in their ability to note differences in real and posed pain given the sex of the emoter. Past research suggests that PFE are recognized differently between sexes, with pain being harder to identify in women’s faces.

Production of jerky from Asian carp meat may increase the consumption of Asian carp in the U.S. Dehydration is a critical step in jerky production. The objective of this study was to determine the effect of dehydration time on the weight loss from jerky prepared from Asian carp meat. Silver carp were harvested from Lake Barkley and deboned through a drum with 3 mm orifice. A total of 1000 g of deboned Asian carp meat was mixed with spices, extruded into 5 mm thick and 2.5 cm jerky strips, and assigned into four treatment groups varying in dehydration times with 6 replicates. The jerky was dehydrated at 145 °F for 3, 4, 5, or 6 hours before they were removed from the dehydrator and cooled to room temperature.  The final weight loss was calculated from the equation (initial weight-final weight)/Initial weight x 100%). Average weight loss ranged from 31% to 48%. As dehydration time increased, weight loss also increased. These results indicate that optimizing the dehydration time is critical as it could affect the jerky yield and its safety

     Non-adults in the archaeological record have been historically overlooked, creating a bias against their social importance. Social age markers are important steps in the transition from non-adult to adult: from childhood to personhood. Theoretical insights concerning social age markers of children describe why non-adults are omitted from the archaeological record. Bioarcheological methods can assist in closing this gap in research by utilizing biocultural components from childhood to determine the implications of life stage transitions. The weaning process is an important period for non-adults, both biologically and socially. The transition from breast-feeding to solid foods can be traced via changes in the isotopic values of carbon, oxygen, and nitrogen, thus providing a timeline for the weaning process. Determining weaning ages through stable isotope analysis has strong implications regarding the concept of personhood in children from past populations since weaning represents the first social age marker in many cultures. While their biological age categorizes them as non-adults, children in the past were often socially considered adults. Combining the social theory of childhood in archaeology and stable isotope analyses, the weaning process, often the first step in the transition from childhood to personhood, can be viewed as a significant social age marker. A cross-cultural investigation of weaning behaviors and sociocultural implications is highly consequential regarding how non-adults are perceived in the archaeological record, the sociocultural identification of ‘children’ in the past, and challenges Western views of ‘childhood’ in the past.  

Adult stem cells are present in many tissues and organs, where they continually generate the specialized cells necessary for tissue function. Stem cells reside in specific microenvironments called “niches” and are regulated by both intrinsic and extrinsic mechanisms. Lipid metabolism has emerged as an important factor influencing stem cell dynamics. Work from our laboratory using the Drosophila melanogaster (Meigen) testis as a model demonstrated that lipid imbalance negatively impacts both germline and cyst stem cells (GSCs and CySCs, respectively). Interestingly, disruption of lipid metabolism in GSCs also led to lipid accumulation in neighboring support cells, suggesting that lipid exchanged between stem and niche cells is important for stem cell maintenance. However, the precise mechanisms that maintain lipid balance under homeostatic conditions remain unknown.

Neural lazarillo (Nlaz), the Drosophila homologue of human apolipoprotein D (ApoD), is a lipid transporter that is expressed in the testis stem cell niche. Preliminary results indicate that depletion of Nlaz in hub support cells causes loss of both hub and stem cells, pointing to a crucial role in niche integrity. Current efforts focus on investigating the mechanisms by which Nlaz regulates niche homeostasis, and whether hub cell loss occurs through cell death or conversion. Given the evolutionary conservation of both Nlaz and ApoD, as well as parallels between fly and human stem cell systems, this work has the potential to uncover conserved lipid-regulatory mechanisms and inform novel therapeutic strategies in regenerative medicine.

Vyjuvek is the first FDA-approved topical gene therapy for dystrophic epidermolysis bullosa, hence referred to as DEB, a devastating genetic skin disorder that causes fragile skin and chronic wounds. DEB is caused by mutations in the COL7A1 gene, which prevents the body from producing type VII collagen, a protein that anchors skin layers together. This treatment addresses this problem at the genetic level by using a harmless, non-replicating herpes simplex virus to deliver a healthy copy of the COL7A1 gene directly to skin cells. When applied as an ointment directly to wound areas, this therapy enables skin cells to produce functional collagen, helping to strengthen skin and support healing. In clinical studies, treated wounds showed significantly improved closure within about 3 months, with durable collagen expression maintained over time, marking a significant advance in patient care. Beyond its immediate benefits for DEB, this approval highlights the growing potential of gene therapy for other rare dermatological diseases with similar genetic origins, such as junctional and simplex epidermolysis bullosa, ichthyosis, and Netherton syndrome. Currently, the annual cost for this therapy is over half a million USD. Future directions will involve reducing treatment costs and expanding gene-based therapies to additional subtypes of epidermolysis bullosa.

Mitochondrial dysfunction is a hallmark of several neurodegenerative diseases, including Alzheimer’s, Parkinson’s, and ALS. The Integrated Stress Response (ISR) is a key adaptive pathway that responds to cellular stress by downregulating global protein synthesis while upregulating stress-responsive proteins such as ATF4. Although the ISR can promote survival, prolonged activation may trigger apoptosis. This study investigates if oligodendrocyte precursor cells (OPCs) and mature oligodendrocytes respond to mitochondrial stress via the ISR. OPCs were treated with various mitochondrial toxins at 8- and 24-hour time points, followed by Western blot analysis. By comparing ISR activation between in-proliferating OPCs and differentiated oligodendrocytes, we aim to determine whether mitochondrial impairment result in the activation of the ISR system. Understanding ISR dynamics in glial cells may offer new insights into therapeutic strategies for neurodegenerative disorders. From the study, it can be determined that impairment of the mitochondrial result in the activation of the Integrate Stress Response in the mature and immature oligodendrocytes.

Highly accurate prediction of functionally relevant structural motifs using deep-learning-based protein structure modeling. 

A fundamental aspect of cellular regulation is the controlled transport of proteins between the nucleus and cytoplasm, governed by nuclear localization signals (NLS) and nuclear export signals (NES). This tightly controlled trafficking ensures proper gene expression, signal transduction, and stress responses. A mutation in either of these signals disrupts cellular regulation, leading to various diseases, including cancers.  NLSs are relatively easy to identify due to their lysine- and arginine-rich sequences. In contrast, NESs are more structurally diverse and harder to determine. To address this, we employed artificial intelligence to model the interactions between proteins and the nuclear export machinery, including CRM1 (exportin), RAN, and RAN-GTP, to predict potential NES motifs. 

Using this approach, we screened more than 2300 human proteins and discovered over 1600 novel NES sequences. We validated our NES discovery using a protein, ANKZF1, which contains a classical NES sequence. Mutation of this NES sequence resulted in constitutive nuclear retention of VMS1, confirming the predicted NES sequence motif. Finally, we also identified a juxtaposed NLS, which suggested a tight co-regulation of NLS-NES motifs. Thus, we successfully applied a deep-learning-based approach to effectively identify and annotate NES sequences, offering a more efficient alternative to traditional sequence-based computational methods. This approach may enhance the understanding of protein transport and contribute to identifying functional motifs relevant to disease.

In humans, a mutation in the CHD7 gene, a chromatin remodeling factor, is closely linked to the congenital disorder CHARGE syndrome. CHARGE syndrome is a developmental disorder that manifests in organs including the eye, ear, and heart. Previous research modeling CHARGE in the mouse recapitulated the haploinsufficient human phenotype and demonstrated embryonic lethality in CHD7 homozygous knockouts. In order to circumvent embryonic mortality, this disorder can be modeled in zebrafish using a CRISPR mutant knockout line. Our previous research using the mouse and fish models was supportive of a role for CHD7 in the development, population number, and mature function of photoreceptors in the retina. The current study aimed to more closely evaluate the effect of CHD7 knockout on the visual acuity of zebrafish larvae. Using our previously validated CRISPR knockout line, we assayed the optokinetic reflex behavior using a well supported behavioral testing paradigm. This experimentation found a statistically significant reduction in the optokinetic reflex behavior of CHD7 maternal zygotic mutant and wild type zebrafish larvae. Maternal zygotic mutants have no functional maternally deposited CHD7 needed to start the developmental program that results in properly formed cone photoreceptors. Although we found a significant difference, more in depth analysis of the optokinetic response behavior via video tracking software and histological corroboration of deficient response with cellular retinal phenotype is necessary to robustly conclude a meaningful acuity deficiency of the maternal zygotic mutants. 

According to the National Cancer Institute’s SEER database, prostate cancer (PCa) is the leading cancer diagnosis among men in the US with over 300,000 men diagnosed each year. It has been observed that African American men in the United States have a significantly higher risk and mortality rate of prostate cancer (PCa) compared to men of Caucasian descent (American Cancer Society, 2024). This difference highlights the importance of investigating novel therapeutic strategies because it reflects underlying issues such as differences in access to healthcare, socioeconomic factors, and tumor biology. Recently it has been observed that African American men with PCa exhibit an increased reliance on de novo lipogenesis for progression and proliferation compared to men of caucasian descent. Our lab utilizes in vitro PCa cell models of both Caucasian (LNCaP) and African American (MDA-PCa-2b) descent to exploit molecular differences and develop novel therapeutic strategies. The focus of this project is to increase the efficacy of the current Docetaxel treatment with the addition of Orlistat, a fatty acid synthase (FASN) inhibitor targeting de novo lipogenesis. Recent publications have indicated a stronger reliance of PCa with African American descent on lipogenesis and may indicate a vulnerability to these kinds of inhibitors (Berchuck, J.E., et al). This project focuses on establishing the standard operating procedure for the lab to perform clonogenic assays, a gold standard in investigating cancer biology. Our results establish the proof of concept for Docetaxel mono treatment that inhibits cell proliferation by microtubule depolymerization. Colony count and intensity were calculated utilizing the ColonyArea plugin with ImageJ software. Future directions involve clonogenic assays in MDA-PCa-2b utilizing Docetaxel and Orlistat mono treatments and in combination to test drug synergy.
 

DNA extraction of Nile Tilapia Oreochromis niloticus (Linnaeus) fin clips using the Promega Wizard® Genomic DNA Purification Kit

DNA extraction of fish is essential for producing intended genetics within aquaculture, as well as informing conservation efforts in our aquatic ecosystems and keeping fisheries sustainable. Four Nile Tilapia fin clips were collected from the anal fin using scissors sterilized in 95% ethanol. A 10-20 mg piece of each fin clip was then added to 600 µl of nuclei lysis solution in a centrifuge tube, which was then combined with 17.5 µl of proteinase K and incubated overnight at 55°C. Three microliters of RNase solution was added to the nuclei lysate, and incubated at 370 C for 30 minutes. Two hundred microliters of protein precipitate solution were added to the lysate, and the solution was chilled on ice for five minutes. Each solution was then centrifuged at 13,000 x g* for four minutes, separating a physical pellet from a liquid supernatant within the centrifuge tube. The supernatant was then removed and added to 600 µl of isopropanol, and centrifuged for 1 minute at 13,000 x g*. The resulting supernatant was removed, and the remaining pellet was treated with 70% ethanol and centrifuged for 1 minute. The resulting supernatant was aspirated, and the remaining DNA pellet was resuspended using the Promega DNA rehydration solution. DNA was quantified using a Nanodrop spectrometer. All DNA samples were found to have a concentration greater than 50 ng/µL, and a 260/280 ratio between 1.8 and 2.0. DNA extraction is an important first step in further genomic analysis such as PCR or sequencing.

Food and fluid intake are influenced by circadian rhythms. A circadian rhythm is a biological variation of rhythms within a cycle of approximately 24 h. Rats are nocturnal, and previous research demonstrates that their ingestive behaviors are primarily conducted during the dark phase. In Spiteri et al. (1981), they found that when food access is restricted to the light phase, rats still consumed the majority of their water intake in the dark phase. This is surprising because food is a primary stimulator of water intake, but it appears the circadian control of water intake is a stronger stimulus than the prandial drive to consume water. This study, however, was limited to males. Studying this in female rats is important because there are well-documented sex differences in ingestive behaviors and circadian rhythms. Therefore, more studies are needed for a complete understanding of how circadian rhythms influence fluid intake, including conducting this research in females. We tested the hypothesis that restricting food access to the diurnal period is not sufficient to shift circadian water intake patterns in male and female rats.  Male and female Long Evans rats (n = 16) were individually housed in modified shoebox cages and were given ad libitum access to water and food except where described. The cages had external lick blocks, allowing for interfacing with the contact lickometer, which allowed for recording of licking behavior.  Rats underwent two weeks of a control period where they were given ad libitum access to food and water. Then, the rats were given food only during the light phase, while having ad libitum access to water for a two-week period. Water intake was measured daily (ml), and licks were measured constantly. Food intake was measured in six-hour bins Monday-Friday and 12-hour bins Saturday and Sunday.

Uniparental Inheritance (UPI) of mitochondria, in which only mitochondria from one sexual parent is passed to offspring, is common in many organisms. For example, Ustilago maydis, a smut fungus that infects corn (Zea mays), passes on mitochondria based in the a locus of haploid parent cells. U. maydis is thought to select the mitochondria with the Rga2 and Lga2 proteins, coded for in cells of the a2 type. Rga2 protects mitochondria from the a2 parent while Lga2 is thought to degrade other mitochondria after successful mating. Interestingly, Sporisorium reilianum (SRZ), another smut fungus closely related to U. maydis, appears to have a similar process with several corresponding homologous genes also located on their a2 locus.

This project explores similarities between the roles of Rga2/Lga2 homologs found in S. reilianum and U. maydis by inserting Rga2 and Lga2 from S. reilianum into the U. maydis model and observing mitochondrial inheritance. By utilizing strains of U. maydis with different mitochondria, namely differing intron lengths in the cox1 gene, we can use PCR techniques to visualize how U. maydis passes on mitochondria with different combinations of native proteins and the S. reilianum homologs. Using this method we found that the S. reilianum Lga2 homolog behaves in the same manner when inserted in U. maydis, but the Rga2 homolog does not.

Diffuse intrinsic pontine glioma (DIPG) is an aggressive, malignant pediatric brain tumor, characterized by mutations of histone H3.3 (H3.3K27M) and TP53, a tumor suppressor gene. The only treatment for DIPG is radiotherapy, which results in a high recurrence rate within 2 years (90% mortality) due to the development of tumor radio-resistance. The hypothesis is that TP53 mutations, at several hotspots such as R175H, R248W, and R273C/H, together with H3.3K27M, drive epigenetics reprogramming leading to radio-resistance and aggressive tumor behavior in DIPG patients. We conducted a transient transfection in HEK293 cells to evaluate the interactions of different combinations of H3.3 and TP53 mutations. Cell proliferation was measured using a luminescent cell viability assay following irradiation. Our results indicated that reintroduction of p53 WT into p53 KO cells rescued the DNA damage response, while specific mutant combinations induced a proliferative advantage, especially with the combination of R273C/H and H3.3K27M. The presence of H3.3K27M enhanced the proliferative effect of the R273H mutant. In conclusion, specific TP53 mutations cooperated with H3.3K27M to promote radio-resistance, suggesting a mechanism of epigenetics interaction.

Ovarian cancer is the deadliest gynecological cancer afflicting women with a 5-year survival rate below 45% and approximately 19,000 new cases diagnosed annually (SEER Database, NCI). Due to the low number of affiliated patients, there is limited focus for drug development in this area.  Metastatic and treatment-resistant ovarian cancers are of particular concern, as they are associated with the poorest survival outcomes. Approximately 80% of patients with advanced-stage disease develop resistance to platinum-based therapies, and response rates to subsequent treatments remain as low as 20%.  To study if our new platinum-based drug will provide a treatment option for patients with cisplatin resistance we aim to develop a drug-resistant cell line. Drug resistant variant cell lines are developed by continual exposure of a specific drug to a parental cell line. Parental cell lines are exposed for a period of 72 hrs. with an IC50 (half maximal drug concentration) and then allowed to recover for 72 hrs.; this continues for a few months depending on the cell line. Cells are then examined for drug resistance through both an MTT and uptake assay. The A2780-Cis cell line was developed through this methodology by continual treatment of cisplatin in the parental A2780 cell line. Based on our preliminary results, the IC50 for our platinum compound in the A2780 cell line is approximately 5µM. Continual exposure will be tested to induce resistance compared to the parental cell line. This is a critical step in determining new treatment options for patients developing cisplatin resistant ovarian cancer.

    Adult or tissue-resident stem cells are an essential population of progenitor cells that are responsible for the regeneration of specialized cells supporting organ function. Upon asymmetric division, stem cells can self-renew and differentiate into specialized cells. Within the male Drosophila melanogaster (Meigen) gonad, three distinct cell populations form a specialized microenvironment known as the “niche”: germline stem cells (GSCs), cyst stem cells (CySCs), and hub cells, which form an organizing center that provides support and maintenance for stem cells. With age, stem cell maintenance and activity decrease due to, in part, the loss of hub cell maintenance and function.

    Changes in acto-myosin contractility can influence the regulation of stem cell niches. However, it is unknown if and how aging may lead to changes in acto-myosin contractility and perhaps niche homeostasis. In our study, we will characterize how acto-myosin forces influence the niche by utilizing Rho1 and Rac1, small GTPases that regulate actin cytoskeleton, and AMP-activated protein kinase (AMPK), a downstream target of actin polymerization. We plan to evaluate how these genes influence hub cell identity and maintenance, and if so, through which mechanism(s). This study has the potential to determine how cytoskeletal signaling pathways interact to preserve hub cell identity and prevent age-related niche deterioration, providing key insights into the mechanistic links between mechanical signaling and stem cell homeostasis during aging.

Cathepsin L is a conserved cysteine protease with roles in development, extracellular matrix (ECM) remodeling, and cancer progression. However, its transcriptional impact during epithelial development remains poorly understood. Using Drosophila melanogaster wing imaginal discs, we investigated how Cathepsin L modulation affects gene expression. The GAL4/UAS system was used to drive both overexpression and RNAi-mediated knockdown, followed by RNA sequencing and bioinformatic analysis. Overexpression altered 581 genes, while knockdown affected 385 genes. Comparative analysis revealed that overexpression and knockdown activated largely distinct transcriptional programs, with only limited overlap. These findings will be discussed and presented. Because of its evolutionary conservation, our findings provide insights into Cathepsin L’s dual role in epithelial regulation and tumorigenesis, offering a foundation for future studies in higher organisms.

     Bovine mastitis, an inflammatory condition affecting the udder tissue of cows, poses a significant threat to the global dairy industry. Triggered by physical injury or bacterial infection, this condition leads to substantial economic losses for farmers. The causative organisms of bovine mastitis are diverse, including both gram-positive and gram-negative bacteria, many of which are highly contagious. The impact of mastitis is significant: it reduces milk production, compromises milk quality, and increases milk discard rates. Additionally, the disease incurs higher treatment and labor costs, often resulting in the premature culling of infected cows. Understanding the cow's innate immune response is crucial for addressing this persistent issue. β-defensins, a vital component of this defense system, serve as the first line of defense against mastitis. However, quantitative analysis of β-defensins in bovine milk is limited in the literature. This study aims to address this knowledge gap by developing a robust analytical method to measure β-defensins in milk. By employing Liquid Chromatography-Mass Spectrometry (LC-MS) coupled with advanced sample preparation techniques, such as size-exclusion and reverse-phase solid-phase extraction, we seek to enhance our understanding of β-defensins. 

Hydrogen has frequently been considered as a potential renewable energy source to replace our current dependence on fossil fuels. An issue with this, however, is the difficulty in storing bulk amounts of a gas without the use of extremely high pressures or cryogenic temperatures. Creating metal hydrides as a solid-state hydrogen storage material has shown promise as a more favorable method of storing hydrogen, with magnesium hydrides in particular being favorable. However, doping the material with metal atoms is necessary for improved hydrogen sorption kinetics in the bulk. Here, multiple structural isomers of small Mg_nTi clusters were first located theoretically using unbiased global optimization techniques.  Once candidate isomers were obtained, we further stringently optimized each structure using the B3PW91 density functional theory method with the 6-311+G(d) all electron basis set for all atoms.  These calculations were performed using the EXPANSE high performance computing cluster at the San Diego Supercomputing Center.  For each determined ground state cluster, electronic properties were further explored and will be presented.  This work builds upon our previous investigation containing only a single magnesium atom in the titanium doped magnesium hydride cluster [1]. Further investigations into the stability of increasing sized clusters and their hydrogen reactivity will provide insight into an optimal hydrogen storage material for a potential future fuel source.

There is growing urgency to develop sustainable alternatives to petroleum-based materials as concerns about environmental impact and resource depletion intensify. Among renewable biopolymers, cellulose stands out as one of the most abundant and versatile organic compounds on Earth. As the structural backbone of plant cell walls, it provides rigidity and mechanical strength, while its abundance, biodegradability, and tunable chemistry make it an attractive candidate for sustainable materials. Beyond its traditional uses in textiles, paper, and food additives, cellulose has been increasingly investigated in polymer chemistry for advanced applications. The present study aims to optimize synthetic routes to dual ionic liquid-functionalized cellulose. Two alternative pathways were evaluated: one beginning with 6-chlorohexanol and the other with caprolactone. Both strategies rely on click chemistry for triazole installation and subsequent triazolium functionalization, but differ in cost, number of steps, and scalability. The goal is to determine which route provides the most efficient balance of yield, labor, and material expense. Characterization of the resulting derivatives was carried out using NMR spectroscopy, infrared spectroscopy, and differential scanning calorimetry to confirm structure and assess thermal behavior. Optimizing the synthetic process will not only advance the design of cellulosic poly(ionic liquids) but also support the broader pursuit of sustainable, high-performance materials.
 

The purpose of this study was to raise awareness about the harmful environmental impacts of traditional plastics and to explore a biodegradable alternative made from milk and vinegar. This experiment aimed to create casein plastic which is a solid byproduct formed when milk curdles with vinegar, and to test its potential as an eco-friendly substitute. To minimize waste, both fresh and expired milk (whole, 2% low fat, and skimmed) were used. After heating the milk, vinegar was added to separate the curds, which were then kneaded, molded, dried, and coated with beeswax to improve water resistance. Each plastic sample was tested for durability by being dropped from shoulder height and for water safety using pH strips. Results showed that plastics made from whole and 2% milk, both fresh and expired, were moldable and durable, while skimmed milk plastics cracked easily. pH levels of the water in contact with the plastics were around 5–6, slightly acidic, indicating some material erosion despite the beeswax coating. Although the casein plastics did not maintain ideal water pH levels, they demonstrated strength, flexibility, and biodegradability. This study suggests that casein plastic, especially from expired milk, can be a sustainable, low-cost alternative to petroleum-based plastics. With further research to enhance water resistance, biodegradable casein plastic could significantly reduce plastic pollution and promote a greener future.

     Wetlands provide essential and beneficial ecosystem services for people as well as benefits for wildlife, but are being rapidly lost. Unused agricultural land can be restored to forested wetlands through manipulation and replanting if hydric soil is present. Forested wetland restorations can struggle due to stunted tree growth and tree mortality, likely caused by intensive agricultural use that alters bacterial community composition and causes nutrient imbalance and loss of organic matter in soil.  

     To assess soil health in restored forested wetlands on agricultural land, soil core samples were collected adjacent to Quercus (oak) trees that were planted in restorations experiencing high tree mortality and stunted growth, in addition to restorations with low mortality and normal growth. Samples were tested for physiochemical properties, such as organic matter and nutrient levels, as well as bacterial functional diversity. Results were compared to score soil health across restoration sites.  

A thermal pre-treatment is required to kill harmful bacteria in beef before dehydration for beef jerky products. The objective of this study was to determine if a thermal pretreatment is also needed before dehydration for jerky prepared from Asian carp meat. Asian carp were harvested from Lake Buckley and deboned through a drum with 3 mm orifice. A total of 1000 g of deboned Asian carp meat was mixed with an equal weight of spices, extruded into strips, and assigned into two treatment groups with 6 replicates: Treatment 1 (cooked at 350 °F for 10 minutes) and Treatment 2 (uncooked). Both treatments were then dehydrated at 145 °F. and samples were analyzed for final weight loss, water activity, moisture content and total aerobic count. Jerky in Treatment 1 had lower final weight, and moisture content than jerky in Treatment 2 group. However, there were no significant differences between the two groups in water activity. There were no detectable bacteria in both groups. These results indicate that dehydrating at 145 °F was able to cook the product and eliminate bacteria from the jerky. A thermal pre-treatment may not be necessary for preparing jerky from Asian carp meat.  

Water activity is important for the safety and stability of jerky products. The objective of this study was to determine the effects of dehydration time on the water activity of jerky prepared from Asian carp meat. Silver carp were harvested from Lake Barkley and deboned through a drum with 3 mm orifice. A total of 1000 g of deboned Asian carp meat was mixed with spices, extruded into 5 mm thick jerky strips, and assigned into four treatment groups varying in dehydration times with 6 replicates. The jerky was dehydrated at 145 °F for 3, 4, 5, or 6 hours before they were removed from the dehydrator and cooled to room temperature.  The water activity of each jerky strip was determined on a water activity analyzer. The water activity ranged from 0.77 to 0.88.  As dehydration time increased, water activity decreased. These results indicate that dehydration time needs to be optimized to achieve a water activity level to assure the safety and quality of jerk prepared from Asian carp meat. 

Salamanders have been used in a variety of different experiments to determine the health of various ecosystems. Biofluorescence was recently studied in amphibians in a study done in 2020. Not much is known about all its functions, more studies are needed to fully understand how it works and how it can be affected by environmental factors. Learning more about the functions of salamander biofluorescence could lead to a greater understanding of how pollutants and human impacts can affect salamanders and the health of their ecosystems. This project aimed to focus on tiger salamander biofluorescence and how its distribution and intensity is affected by the exposure of natural sunlight. Unmarked, 2nd year larval salamanders were taken from a single diverse pond to control for any differences among the ponds. The larvae were exposed to three different light treatments being dark, dimmed, and light. Within these light treatments there were also two time treatments that were six and twelve days. Their biofluorescence was photographed before and after the time treatments with a blue excitation light and a 500 nm long-pass filter. All images were quantified using ImageJ. The results from this experiment support the idea that salamander biofluorescence may be partly environmentally dependent and this could provide useful information for future studies to further understand the functions of amphibian biofluorescence. Furthermore, more extensive research into this topic could provide a new indicator of ecosystem health for use in conservation.

Maternal mortality, or the death of a woman during pregnancy or childbirth, is still a serious health problem around the world. Many women die from preventable medical causes like heavy bleeding, high blood pressure, infections, and unsafe abortions, usually during labor or shortly after giving birth. But these medical causes are not the whole story. Social issues such as poverty, gender inequality, poor education, racism, and weak health systems play a significant role. In rural areas, like Southcentral Kentucky, mothers often face long travel distances, few doctors, and limited hospital resources, which makes pregnancy and delivery more dangerous. In this study, we will use a meta-analysis approach to identify the causes of maternal deaths that are due to healthcare access problems and to suggest possible solutions. We are conducting several analyses to find out why Kentucky has one of the highest maternal mortality rates in the country. Findings indicate that many women experience birth-related trauma due to factors such as a lack of patient-centered care, medical interventions, and insufficient support. These findings call for changes in healthcare practices to reduce the maternal mortality rates in the state of Kentucky.  

Migraines affect over one billion people worldwide, with current management approaches largely reactive rather than preventive due to the difficulty of predicting onset. This study develops a real-time migraine prediction model integrating blood oxygen saturation (SpO₂) and barometric pressure data. The model's predictive performance achieved 94.50% accuracy, 0.972 AUC-ROC (area under the receiver operating characteristic curve, indicating excellent discriminative ability), 0.889 precision, and 0.821 recall. Model reliability was verified through multiple validation approaches: five-fold cross-validation demonstrated exceptional stability (coefficient 0.0075), while a small training-test performance gap (0.0345) confirmed the model wasn't overfitting to training data. Statistical validation through proportions z-test (z=8.684, p<0.001, null accuracy=0.803) verified performance significantly exceeded chance. SHAP (SHapley Additive exPlanations) analysis provided model interpretability, identifying experimental hypoxia-triggered attacks (44.3%) and hypoxia-induced headache (17.7%) as the strongest predictors. The model architecture, selected after comparative analysis of six machine learning approaches, employs an optimized Random Forest classifier incorporating clinically-established SpO₂ thresholds and barometric pressure changes known to trigger migraines. The algorithm's reliance on data that can be extracted from standard smartwatch sensors and environmental measurements makes it particularly suitable for integration into existing wearable health monitoring systems, enabling a shift from reactive to preventive migraine care, with potential benefits for millions of migraine sufferers.

As the obesity crisis expands across developing countries, the prevalence of obstructive sleep apnea has risen dramatically. This chronic condition involves periodic episodes of hypoxia and sleep fragmentation (SF), which trigger inflammatory responses in the brain and peripheral tissues. Most studies evaluating sleep fragmentation have focused on males, though females are also affected. Given known sex differences in immune function, we examined the inflammatory responses of female and male C57BL/6J mice following varying durations of chronic sleep fragmentation (CSF; 1, 2, 4, and 8 weeks) or control conditions with no sleep fragmentation (NSF). Sleep disruption was induced using an automated cage system in which a sweeping bar moved across the cage floor every 2 minutes for 12 h/day (8 a.m.–8 p.m.). Brains were collected, and the hypothalamus, hippocampus, and prefrontal cortex were dissected for analysis of interleukin-1β (IL-1β) and tumor necrosis factor-alpha (TNF-α) gene expression via RT-PCR.
In the hypothalamus, males under CSF exhibited a significant TNF-α peak at Week 4, whereas females showed no major differences between NSF and CSF conditions. Males also demonstrated distinct IL-1β peaks under CSF, indicating heightened inflammatory sensitivity. In hippocampus, females displayed a strong, short-lived surge in both cytokines under CSF, while males showed faster resolution of inflammation compared to controls. In the prefrontal cortex, females exhibited dramatic, highly variable TNF-α and IL-1β spikes under CSF, whereas males displayed more moderate, transient increases.
These results demonstrate that chronic sleep fragmentation induces region- and sex-specific neuroinflammatory responses, revealing differential inflammatory responses between male and female mice.

Unlike in the central nervous system, tissues of the peripheral nervous system have the capacity to regenerate after injury. This unique ability comes from a complex collaboration between multiple cell types to repair injured nerve tissues and ultimately restore their function. Part of this repair involves axon and myelin degeneration of the nerve segment distal to the injury site (Wallerian degeneration), a process mediated by dedifferentiated (repair) Schwann cells and the immune system. Revascularization is also required to reestablish function of the nerve. This study aims first to visualize immune and vascular changes after a full transection injury at the tissue level using immunohistochemical (IHC) analysis. Secondly, it will identify key transcripts and proteins implicated in the immune, vascular, and other systems’ responses to injury via transcriptomics, proteomics, and pathway analyses. As expected, IHC and differential expression analyses provide evidence of neovascularization and immune activation 15 days after injury, suggesting roles in the degeneration and subsequent healing process of the nerve. Further, analysis of differentially expressed genes between the intact and injured condition show that a direct relationship exists between expression of many transcripts and their corresponding proteins. Finally, functional pathway analysis using Reactome revealed the cell cycle as the most upregulated group of pathways 15 days after injury. Cell senescence, DNA modification and repair, and EGFR signaling were also upregulated and myelination and apoptosis pathways were downregulated. Our findings are among the first experimental evidences revealing subacute pro-regenerative changes during Wallerian degeneration in the transected adult human nerve.

On average cost billed to patients for 1 dose of Alteplase ranged around $8434. Normal dosing of TPA requires 0.9mg/kg (not to exceed 90mg). This medication is the difference between lifelong disabilities as well as death. If you find your self in the emergency room being treated for your unplanned stroke, are you going to refuse the one medication that will save the neurons in your brain from death? The obvious answer is no. As one could imagine, spending an unplanned $8400 can be detrimental for people. In 2023, the median Kentucky income per individual was $34,030 per year. Most seen today is the production of TPA via Chinese hamster ovary cells. This method is both expensive and inconvenient. My research at Pikeville University is aiming to raise production rates and lower treatment costs around Appalachia for TPA. We have begun using plant-based platforms such as Tobacco plants to produce TPA. By using this technology, we can eliminate both virus risk and cost implications that are associated with the use of hamster cells. This research can reduce the cost of TPA to a bare minimum by 50%. This is a significant step in leading accessibility of TPA around both the US and Appalachia.

Billions of birds migrate every spring and fall to reach their breeding or wintering grounds. Several populations live in sympatry on the wintering grounds but breed at different latitudes. The dark-eyed junco (Junco hyemalis) is one such species. Migrants breeding at higher latitudes typically respond to early spring day length by increasing fat stores to prepare for long-distance migration, while resident populations initiate breeding earlier at lower latitudes. Very limited information is available about how gut microbial communities may contribute to digestion, immunity, and energy storage during this preparation period. To investigate whether gut microbiome differences contribute to these seasonal physiological changes, we collected gut tissue samples from both resident (J. h. carolinensis) and migrant (J. h. hyemalis) juncos across three life-history stages: fall (non-breeding), spring (resident breeding initiation and migrant pre-migratory fattening), and summer (resident breeding at lower latitudes and migrants at higher latitudes). DNA and RNA were extracted using the ZymoBIOMICS DNA/RNA Miniprep Kit, and Nanodrop spectrophotometry confirmed high-quality nucleic acids for sequencing. Current results show that migrants and residents differ in fat accumulation patterns despite shared environments, suggesting that gut microbial communities, along with photoperiod cues, may influence seasonal fat deposition. These findings support future microbiome sequencing to better understand the role of microbial variation in migratory physiology and seasonal adaptation.

In this project, we are studying the oscillations of a rolling cylindrical container filled with fluids of varying viscosities on an isochrone (Tautochrone) ramp. The goal of the project is to determine how the oscillation period changes depending on the viscosity of the fluid inside the container, and develop a data acquisition system (DAQ) to record video data. Coordinates of the cylinder while oscillating were extracted from two videos from a stereoscopic camera system connected to a raspberry PI 4. We developed a DAQ system using python witch tracks the cylinders center and determines its position, velocity, and acceleration. In this presentation, we will discuss the developed experimental setup, present the data we have collected with various viscosities, and discuss the software developed to determine cylinder position.

Neural networks are a powerful tool for solving complex classification problems. This research applies them to algorithmically classify different structural types of simulated polymers. Two datasets were used: one consisting of descriptive structural parameters selected by the researchers, and another containing the raw three-dimensional coordinates of the polymer. Both approaches performed at a high level on the test data, suggesting that unprocessed coordinate data can be used effectively in the future. This simplification may streamline analyses, reduce preprocessing time, and enable unstructured learning approaches.

ATP synthase is a key membrane protein that produces adenosine triphosphate (ATP), a vital energy source for all living organisms. It uses energy from the movement of protons across the membrane to produce ATP. It can also break down ATP to ADP and move protons against the regular flow. In this project, we prepared E. coli ATP synthase membrane vesicles with the cytoplasmic part of the ATP synthase exposed to the exterior, making it easier to test its function. We made the inside-out membrane vesicles, measured them using a modified Lowry assay, and performed a hydrolysis assay to evaluate the ATP synthase activity.