Forensic entomology is an important field of forensic science that utilizes insect evidence in criminal investigations. Blow flies (Diptera: Calliphoridae) are among the first colonizers of remains and are therefore frequently used in determining the minimum postmortem interval (mPMI). Blow fly development, however, is influenced by a variety of factors including temperature and feeding substrate type. Unfortunately, dietary fat content remains an understudied factor on the development process, which is problematic given the relatively high rates of obesity in the United States. To study the effects of fat content on blow fly development we investigated the survivorship, adult weight and development of Lucilia sericata (Meigen; Diptera: Calliphoridae) and Phormia regina (Meigen; Diptera: Calliphoridae) on ground beef with a 10%, 20%, or 27% fat content. As fat content increased, survivorship decreased across both species with P. regina being significantly impacted. While P. regina adults were generally larger than L. sericata across all fat levels, only L. sericata demonstrated a significant (P < 0.05) difference in weight by sex. Average total development times for P. regina are comparable to averages published in other literature. Average total development times for L. sericata, however, were nearly 50 hours higher. These findings provide insight on the effect of fat content on blow fly development, a factor that should be considered when estimating a mPMI. By understanding how fat levels affect the survivorship and development of the species studied here, we can begin improving the practice of insect evidence analysis in casework.

Medicolegal forensic entomology is the study of insects to aid with legal investigations (Gemmellaro, 2017). Insect evidence can be used to provide information such as the post-mortem interval (PMI). Blow flies are especially useful as these insects are primary colonizers, quickly arriving at a corpse (Malainey & Anderson, 2020). The age of blow flies found at a scene is used to calculate the PMI. Blow fly age can be estimated using weather data as these insects are poikilothermic (Okpara, 2018). Morphological analysis also can be used to estimate age; however, it is more difficult with pupal samples as the pupae exterior does not change significantly as development progresses (Bala & Sharma, 2016). Gene regulation analysis can estimate the age of samples. MicroRNAs are short noncoding RNA that regulate gene expression (Cannell et al., 2008). Here, we aim to catalog miRNAs expressed during the development of three forensically relevant blow fly species preserved in several storage conditions. Results demonstrated that various miRNA sequences were differentially expressed across pupation. Expression of miR92b increased during mid pupation, aga-miR-92b expression increased during early pupation, and bantam, miR957, and dana-bantam-RA expression increased during late pupation. These results suggest that microRNA can be used to estimate the age of pupal samples as miRNA expression changes throughout pupation. Future work could develop a statistical model to accurately determine age using miRNA expression patterns.

Triple Negative Breast Cancer (TNBC), indicated by the absence of estrogen, progesterone and human epidermal growth factor receptor 2 (HER2), is the most aggressive form of breast cancer characterized by high rates of metastasis and low survival. Among those diagnosed with TNBC, 34% contain Inhibitor of Growth 4 (ING4) deletion that is associated with poor patient outcomes. We previously showed that ING4 negatively regulates NF-B in breast cancer. Previous studies show parthenolide, a compound found in feverfew (Tanacetum parthenium) to inhibit NF-B in cervical and gastric cancer. We hypothesized that parthenolide inhibits cytokine-induced activation of NF-B in ING4 deficient TNBC cells. To test the hypothesis, previously established vectors, v2, ING4 wildtype and v2h1, ING4-deleted were synthesized in MDA-MB 231, a TNBC cell line, using a CRISPR/Cas9 system. Inflammatory cytokines, IL-1 and TNF, were tested in ING4 wildtype or ING4 deleted cells for elicited phosphorylation of NF-B, proliferation, and migration in the presence or absence of parthenolide. The results showed that TNF or IL-1 induced translocation phosphorylation of NF-B regardless of ING4 deletion. ING4 inhibited proinflammatory cytokine induced pp65, consistent with previous studies demonstrating the negative regulation of NF-B in ING4-sufficent cell lines. We found the optimal working dose of parthenolide, 100nM, had no effect on cell proliferation in the presence or absence of IL-1. Parthenolide inhibited IL-1induced phosphorylation of NF-B regardless of ING4 deletion. Parthenolide inhibited TNF-induced phosphorylation of NF-B in ING4-deleted cell lines. Moreover, parthenolide induced migration of TNBC cells regardless of ING4 presence of absence. TNF and parthenolide treated samples in ING4-deleted cell lines were found to inhibit cell migration to basal level. These results demonstrate the difference in inhibitory mechanism of parthenolide in induced phosphorylation of NF-B through proinflammatory cytokines TNF or IL-1This is demonstrated by the exclusivity of parthenolide inhibition of TNF induced phosphorylation of NF-B in ING4-deleted TNBC cell line. In contrast, parthenolide inhibition of IL-1 induced phosphorylation of NF-B occurred regardless of ING4 deletion. These results may inhibit parthenolide as an alternative to those with ING4-deleted TNBC due to its role in inducing cancer phenotype cell migration.

The ever-increasing importance of cancer and neurodegenerative diseases continues to grow as populations across the world are affected by death and aging. The vitamin A (RXR) and vitamin D (VDR) receptor pathways offer promising potential to aid in treatment of cancer and Alzheimer’s disease. This thesis discusses the potential application of novel analogs of Bexarotene (RXR agonist), MeTC7 (a new potent VDR antagonist), and vitamin D as possible therapeutics for cancer and Alzheimer’s disease.

The field of forensic science has been growing and changing with improvements in DNA analysis. One field affected is forensic entomology, which is exploring many ways in which DNA can increase the application of insects in forensic science. One application being explored is the use of insects as a source of human DNA in a criminal investigation. Using flies as a source of foreign DNA can also be utilized in ecological research to conduct surveys on the various species present in different environments. This experiment intends to determine if flies can act as a viable source of alternate DNA. This will be accomplished by an ecological survey of DNA extracted from flies. DNA extractions were performed on flies gathered from parts of the greater Phoenix area. The DNA was then amplified with primers targeting different animal species and examined to observe what animals the flies had come in contact with. Several samples had contamination due to human error and were not able to be evaluated. One DNA extraction out of fifteen yielded pig DNA, indicating flies can be used as a source of DNA. Future experiments should use different animal primers and amplify sections of DNA that can determine the different species consumed by flies. Further research into flies as a DNA source can increase the amount of information available to forensic scientists as well as improve ecologist’s observation of an environment’s biodiversity.