The study examines two naphthoquinone compounds with similar structures but differing toxicities, menadione and plumbagin. Menadione is a synthetic derivative of Vitamin K, while plumbagin is a plant-derived toxin used in homeopathic medicine for its beneficial qualities. Both compounds have demonstrated anti-cancer ability and the ability to produce reactive oxygen species (ROS), but it is unclear from current literature what causes the differences in toxicity. It was hypothesized that plumbagin may produce relatively more ROS or more potent ROS when compared to menadione under various conditions, thus leading to increased toxicity. The study aims to characterize these compounds by examining their ability to cleave DNA and produce ROS. Menadione did not produce significant chemical cleavage under any conditions studied, while plumbagin does at 1 mM with 1 mM sodium ascorbate. Photocleavage showed the opposite effect, with menadione producing more cleavage compared to plumbagin, with an average of 83% and 45% respectively. An ROS probe assay utilizing a fluorescence microplate reader and fluorescein standard was developed for use in this study based on previous literature. Menadione and plumbagin have been examined for hydroxyl radical production using the ROS fluorescent probe HPF. The assay revealed that both compounds produce hydroxyl radicals in a concentration-dependent manner, which could explain the results from photocleavage experiments. Similar analyses will be performed to examine other ROS probes such as Singlet Oxygen Sensor Green. Both compounds will also be examined for differences in DNA cleavage and ROS production in vivo using E. coli.

Flaviviruses pose a significant global health threat, causing diseases that range from mild flu-like symptoms to severe hemorrhagic and neurological diseases. These viruses are typically spread by ticks or mosquito bites. Usutu virus (USUV) is a flavivirus currently circulating in Europe, with the potential to invade the central nervous system in humans. While neuroinvasive disease due to USUV is rare, it can result in severe conditions such as encephalitis, meningitis, and encephalomeningitis. This study aims to characterize USUV replication from peripheral sites through the central nervous system using in vivo mouse models, specifically C57BL/6 Wild-Type (WT) mice, type I interferon-deficient (IFNAR1-/-) mice, and type I and II interferon-deficient (AG129) mice. Our results showed that IFNAR1-/- and AG129 mice are highly susceptible to USUV infection, resulting in 100% mortality. WT mice exhibited lower viral replication in peripheral organs compared to IFNAR1-/- and AG129 mice. In addition, flow cytometry data showed high induction of T cell and B cell immune response along with innate immune response in all three mouse models. Additionally, immune responses were analyzed through qRT-PCR and multiplex immunoassay to observe transcriptional and translational changes following USUV infection. In conclusion, the data obtained from this study provides valuable insights into the viral replication kinetics and immune responses to USUV infection in vivo.

Sodium Chloride Enhances the Photo-Toxic Properties of a 1,4-Naphthoquinone Found in Marine Environments Bianca Vazquez, Owen Waters, Karen Amike, Sneha Kannan, Kathryn B. Grant Naphthoquinones are released into the environment through the incomplete combustion of wood, coal, petroleum, and other organic fuels, especially diesel. A number of naphthoquinones are not only highly toxic to humans but also to organisms that inhabit soil and aquatic environments. Through rainfall, these compounds find their way into bodies of water where they can cause great harm. By sensitizing the production of DNA damaging reactive oxygen species (ROS), irradiation of the PAHs by sunlight often dramatically enhances their toxicity to a variety of life forms. In investigating this problem, our laboratory discovered that high sodium chloride (NaCl) concentrations similar to those that exist in marine environments dramatically increase the levels of dangerous ROS generated by PAHs with anthraquinone frameworks consisting of three fused aromatic rings. The goal of the present study is to determine if naphthoquinone PAHs can also become more phototoxic when high NaCl concentrations are present. Because naphthoquinones have only two aromatic rings, they are more water soluble and therefore are observed at higher concentrations in marine environments. UV-visible spectroscopy was used to determine the stability of the compounds in dimethyl sulfoxide (DMSO) and in sodium phosphate buffer. Additionally, the MDID-5 Plate Reader was utilized to assess the fluorescence of each compound upon irradiation with and without NaCl. These results offer insight into potential levels of hydroxyl radicals generated by each compound via detection by hydroxyphenyl fluorescein (HPF). In addition to affording important mechanistic insights, it is hoped that our work will uncover the structural features that enable PAHs to display a salt-induced ROS enhancement.

INTRODUCTION: Older Black Americans are at a much higher risk for Alzheimer’s Disease (AD) than Caucasian counterparts, a difference which can only partially be explained by socioeconomic factors and other medical information. In this study, we investigated cerebrospinal fluid (CSF) biomarkers and their predictive role on subcortical atrophy in the precuneus and posterior cingulate in a middle aged, at-risk, racially diverse cohort. METHODS: The data set included 76 cognitively unimpaired, middle aged, Black Americans (N=29, F/M=17/12) and Non-Hispanic White (N=47, F/M=27/20) participants. We compared CSF tau, Aβ42, Aβ40, precuneal volumes, and posterior cingulate volumes. RESULTS: Brain volume analysis revealed that Black American men were found to have the lowest precuneal volumes of all groups compared, with white participants not demonstrating any sex differences in precuneal volumes. CSF biomarker analysis shows that Black American participants often have lower CSF Aβ42, which is related to lower precuneal volumes, a correlation that was not noted in the groups of white participants. DISCUSSION: Results make sense considering previous literature, and because lower CSF Aβ42 is indicative of higher Aβ plaque burden, our results indicate that in Black Americans, CSF Aβ42 could be utilized to predict level of Aβ plaque burden and resulting subcortical atrophy of the precuneus. This research is important because not all measurable differences across race can be attributed to socioeconomic factors like ADI. In the future, comparisons should be made with larger sample sizes and including information about cardiovascular risk factors and white matter hyperintensities.

Adipocytes enlarge in response to high-nutrient balance, such as in the western diet, which stimulates recruitment of macrophages and other immune signaling proteins promoting apoptosis, cell death. Blueberries (BB) contain bioactive compounds that have been reported to favorably modulate the apoptosis signaling cascade. This investigation aimed to examine the effects of BB consumption on mechanisms of adipose tissue apoptosis in mice fed a western-style diet. UM-HET3 male mice were fed either a low-fat, low-sugar, low-salt (LFLSS; n=8), a high-fat, high-sugar, high salt (HFHSS; n=7) or HFHSS combined with 5% w/w freeze-dried BB (HFHSS + BB; n=8) diets for 12 weeks. Whole epidydimal fat pads (eWAT) were collected at sacrifice and Western Blot was used to assess proteins implicated in the intrinsic apoptosis signaling pathway such as B-cell lymphoma-extra-large (BCL-XL), BAX (Bcl-2-associated X protein), caspase-9, and monocyte chemoattractant protein (MCP)-1. There were no differences in absolute or relative eWAT mass (p = 0.5). Compared with LFLSS, there was an increase in BCL-XL expression in HFHSS + BB (1.00  0.35 vs. 1.98  0.71; p = 0.02). The expression of caspase-9 was attenuated in HFHSS + BB compared to both LFLSS and HFHSS, albeit not significantly. No significant differences were observed in the expression of BAX, and MCP-1. Preliminary findings indicate a promising role for BB to protect adipocytes against apoptosis induced by a western diet contributing to overall metabolic and cellular resilience. This may prevent the development and/or delay the progression of metabolic disorders. Further analysis is underway.