Hyperglycemia is a key risk factor for metabolic disorders. Western diet, rich in fats and sugars, increases the risk of metabolic disorders and oxidative stress. Blueberry (BB) polyphenols may help manage hyperglycemia via their antioxidant properties. Therefore, this study examines the impact of BB supplementation on glucose metabolism and hepatic oxidative stress in mice consuming a Western diet. Four-week-old UM-HET3 male mice were assigned to a control (low-fat, low-sucrose, low-salt, LFLSS) alone or supplemented with 5% BB (LFLSS + BB) for four weeks. Then, LFLSS either continued with that intervention or were randomized to HFHSS while LFLSS + BB was switched to HFHSS + BB (5% w/w) for 12 weeks. Insulin (ITT) and glucose tolerance tests (GTT) were performed at weeks 14 and 15, respectively. Liver was collected at sacrifice for protein expression analysis of oxidative stress markers by Western Blot. Normality was examined using Shapiro-Wilk test. Data were analyzed using one-way ANOVA or Kruskal-Wallis followed by the appropriate posthoc test. No significant difference was observed for ITT or GTT across all the groups. HFHSS decreased expression of the antioxidant enzymes glutathione peroxidase (GPx)-1, GPx-3 and superoxide dismutase (SOD)2, though not significantly. BB significantly increased GPx-1 expression compared to the HFHSS group but had no effect on GPx-3. BB numerically attenuated the HFHSS-induced decrease in SOD2, however. Our preliminary findings indicate that consuming a BB-rich diet may attenuate hepatic oxidative stress. Further analysis is ongoing.

Neurotropic flaviviruses continue to spread and cause human disease in new areas of the world. Powassan virus (POWV) causes encephalitis, including fatal neuroinvasive disease in approximately 10% of cases, and long-lasting neurological sequelae in 50% of survivors. Zika virus (ZIKV) infections have caused a wide spectrum of neurological diseases, such as Guillain-Barré syndrome, myelitis, meningoencephalitis, and congenital microcephaly. MicroRNAs (miRNAs) are a group of small RNAs involved in the regulation of a wide variety of cellular and physiological processes. To identify differentially expressed miRNAs associated with POWV and ZIKV infections, we employed nCounter® Technology (NanoString) to analyze global miRNAs expression in mice brains and primary mouse cortical neurons. Our data demonstrate that POWV and ZIKV infections cause global downregulation of miRNAs with only few up-regulated miRNAs. ZIKV infection also results in downregulation of miRNA processing enzymes including Dicer-1, Drosha, DGCR8, AGO1 and AGO2 in neurons. Biofunctional analysis revealed that POWV- and ZIKV-modulated miRNAs regulate the pathways related to neurological development and neuroinflammatory responses. Moreover, functional studies targeting specific miRNA revealed a critical role of miR-155 in the pathogenesis of POWV and ZIKV infection. Primary mouse cells lacking miR-155 were more susceptible to infection with POWV and ZIKV compared to cells derived from the wild-type mice. Collectively, these data indicate that cellular miRNAs may play a critical role in the pathogenesis of neurotropic flavivirus infection.

Abstract- Workplace violence (WPV) is a critical issue in healthcare, with Type 2 WPV, violence from patients or visitors directed at healthcare workers, being the most prevalent. Registered nurses (RNs) frequently experience verbal and physical assaults, leading to increased stress, burnout, and reduced job satisfaction, a problem exacerbated by the COVID-19 pandemic. Effective interventions to mitigate WPV and protect nurses are essential to ensure safer healthcare environments. This project aims to assess whether de-escalation training can improve nurses' confidence in handling patient aggression, evaluating the impact of training on RNs' ability to manage violent situations as measured by the Confidence in Coping with Patient Aggression Instrument (CCPAI). A pretest-posttest design will be used, targeting registered nurses in Georgia and New York. Participants will complete the CCPAI to measure their confidence in managing aggression before and after receiving de-escalation training, which focused on communication strategies, non-threatening body language, and techniques for maintaining calm during aggressive encounters. Paired t-tests and descriptive statistics will be used to analyze the data, comparing confidence levels pre- and post-training. Results are pending. The study’s findings will inform future strategies to address workplace violence and enhance safety in healthcare settings.

Gordon Holmes Syndrome (GHS) is a neurodegenerative disorder associated with mutations in the E3 ubiquitin ligase RNF216, leading to a range of neuroendocrine, cognitive, and motor impairments. Disruptions in the hypothalamic-pituitary-gonadal axis have been identified in Rnf216 knockout (KO) mice, suggesting a role for RNF216 in neuroendocrine function. To determine if RNF216 regulates cognitive and motor functions, we performed motor and learning tasks in adult and middle-aged male and female Rnf216 KO and Wildtype (WT) mice. Although no significant motor phenotypes were observed, female KO mice exhibited abnormal limb clasping. Both sexes displayed age-dependent deficits in strategy and associative learning. We next examined microglia density, area, soma size, and morphology in multiple brain regions of Rnf216 KO mice. Our results revealed sex- and region-specific changes: female KO mice exhibited increased microglia density in the hippocampus and smaller soma size, while male KO mice had reduced microglia intersections in the cortex. We also observed unexpected changes in lateral ventricular size in Rnf216 KO mice. Our findings reveal neurobehavioral, structural, and microglial alterations in Rnf216 KO mice. Future experiments will use conditional knockout strategies to determine if selective deletion of Rnf216 in microglia causes similar phenotypes to Rnf216 global KO mice.

Donor–π-acceptor (D–π-A) fluorophores are organic molecules that consist of a donor unit, a conjugated π-bridge, and an acceptor unit. This structure allows for efficient intramolecular charge transfer (ICT) between the donor and acceptor through the π-bridge, resulting in strong fluorescence emission and customizable optical properties. By altering the donor, acceptor, or π-bridge, we can fine-tune the fluorophores' quantum yields, chemical properties, and emission wavelengths, which can range from ultraviolet-visible (UV-Vis) light (400–650 nm) to near-infrared (NIR) regions, including NIR-I (650–900 nm) and NIR-II (1000–1500 nm). These fluorophores are of great interest due to their unique photophysical characteristics, making them promising candidates for biomedical and bioimaging applications. This research presents the synthesis, characterization, and photochemical properties of various D–π-A analogs composed of malonic acid and indanedione derivatives as the acceptor and various indole derivatives, connected by a Vilsmeier-Haack linker. This work builds on ongoing studies in our laboratory aimed at enhancing the properties of donor–π-acceptor fluorophores for future bioimaging applications

Increased intestinal permeability (IP) is associated with high blood pressure (BP). Blackberries (BL), blueberries (BB), raspberries (RB), and strawberries (SB) are widely studied for their anti-inflammatory and antioxidant effects and shown to preserve intestinal barrier function. Thus, the objective of this study was to evaluate the effects of a berry-rich diet on BP and IP in angiotensin (Ang) II-treated rats. Eight-week-old male Sprague-Dawley rats were fed an AIN-93M diet (control, Ang II, and Losartan (LOS; 20 mg/kg/BW)) or AIN-93M diet supplemented with 10% w/w freeze-dried berries (equal parts BL, BB, RB, and SB; Berry) for 8 weeks. Rats were implanted with subcutaneous osmotic minipumps delivering 0.9% saline (CON) or Ang II (270 ng/kg/BW/min; Ang II, LOS, and Berry) at week 4. BP was measured weekly. IP was measured by FITC-dextran 4K assay. Protein expression of inflammation, and oxidative stress in the proximal colon were assessed using Western Blot. Berry supplementation attenuated increases in BP after 8 weeks. IP was reduced in Berry group compared to Ang II, albeit not significantly. Protein expression of MYD88, an adaptor protein whose dysregulation may cause chronic inflammation, increased significantly in all groups. While the antioxidant enzyme, catalase, decreased in all groups, Berry preserved GPx1 expression. We demonstrate that a berry-rich diet protects against increases in BP and IP in Ang II-treated rats which may be through preservation of antioxidant enzymes. Future analyses will investigate tight junction gene expression and colon fibrosis.

Autoimmune diseases are diverse disorders arising from abnormal cellular and humoral responses to self-antigens in the immune system. However, the underlying cause of autoimmunity remains unclear. In this study, ready-made autoreactive CD4+ T cell hybridomas (containing autoreactive CD4+ lymphocytes from scurfy TCRmini mice with BWα−β− thymoma line) were cloned and their antigen receptors (TCRs) were characterized to decipher the specificity of autoreactive TCRs. This approach was used to understand the mechanisms of autoreactivity in the immune system and assist in redirecting abnormal CD4+ T cells present in the peripheral repertoire of healthy individuals for therapeutic advantage. Some hybridoma clones expressed CD4+ and Vα2+ TCRs with unique amino acid sequences and they upregulated Nur77GFP expression upon autologous DCs stimulation and a stronger expression of Nur77GFP in the presence of aCD3 stimulation. Our data suggests that these activated hybridoma clones TCRs have an elevated binding affinity for self-peptides/MHCII complex and are likely autoreactive. INDEX WORDS: autoreactive CD4+ T cell hybridomas, Vα2+ TCR, Nur77GFP, autologous DCs, antiCD3, scurfy TCRmini mice, BWα−β−thymoma line

Pseudomonas aeruginosa is a pathogenic bacterium causing human infections. The bacterium relies heavily on D-2-hydroxyglutarate dehydrogenase for L-serine biosynthesis, making the enzyme a potential target for antibacterial strategies. PaD2HGDH, a Zn2+ and FAD-dependent enzyme, catalyzes the oxidation of D-2-hydroxyglutarate or D-malate to 2-ketoglutarate or oxaloacetate. Metal substitutions of Zn2+ with Cd2+, Co2+, Mn2+, and Ni2+ have been proven to maintain the enzyme's activity, but their effects on the kinetic mechanism of the enzyme are still unclear. In this study, recombinant His-tag bacterial enzyme was purified to high levels with Cd2+, Co2+, Mn2+, Ni2+, and Zn2+ to investigate the effects of the various metals on enzyme catalysis. Irrespective of the metal bound to the bacterial enzyme, 1 mM phenazine methosulfate (PMS), the artificial electron acceptor used in the enzyme kinetic assays, was saturating for all the metal-loaded enzymes with D-malate as substrate. Viscosity studies with glucose were conducted to understand the effects of the various metals on the bacterial enzyme rate-limiting steps at pH 7.4 and 25°C. The viscosity effects on the kcat parameter showed that while the turnover rates of the Co2+, Mn2+, and Zn2+ loaded enzymes were limited by both catalysis and diffusional steps, with slopes of 0.13, 0.47, and 0.52, respectively, the Cd2+ (slope of 1) and Ni2+ (slope of 0) loaded enzymes were fully limited by diffusion and catalysis, respectively. Similar results were obtained when the viscosity effects on the kcat/Km parameter were investigated. The data demonstrate that loading the enzyme with various metals alters the rate-limiting steps during enzyme catalysis with D-malate.

Severe acute respiratory syndrome coronavirus 2 (SARSCoV- 2), which infects mainly the respiratory tract, caused a worldwide pandemic of the coronavirus disease 2019 (COVID-19) from 2020 until 2023. The emergence of new mutations is linked to consequent progressive selection of different dominate variants of concern (VOCs) (Wuhan-Hu-1 Wildtype (WT), Alpha, Delta, Omicron) and changes in viral properties. Presently, Omicron is the predominant variant, which is 2 to 4 times more transmissible and has an increased immune evasion compared to other variants. As infections continue, SARS-CoV-2 may further evolve to better evade the human immune system, such as the humoral immune system, and/or boost transmissibility. So far, existing vaccines target the highly, mutagenic spike protein (S), which allows viral entry into host epithelial cells via ACE2 receptor binding and is the main target for neutralizing antibodies (nAbs). To limit virus replication and transmission of SARS-CoV-2, a greater T-cell response is needed to eliminate virus infected cells at a more rapid rate. T-cell responses play a critical role in vaccine protection against severe COVID-19 disease by contributing to clear the infection and aiding in memory B cell activation for long-term protection. A way to increase cellular immune response is by incorporating the nucleocapsid (N) protein in vaccine development which is: 1) highly conserved amongst Sarbecovirus strains; 2) highly immunogenic; 3) abundantly expressed; and 4) interacts with various host cellular processes in the cellular immune response like inducing strong T cell responses. Thus, including a higher T cell response in the host may result in lower viral titers by eliminating virus-infected cells at a more rapid rate. The candidate vaccine will include: 1) the S protein of SARS-CoV- 2 VOC to induce the greatest nAb response to help counteract infection; 2) the N protein (highly, conserved) of SARS-CoV-2 VOC that elicits T-cell responses.

While using one’s voice is a habituated motor skill, intentionally altering one’s voice quality requires mental effort. Although elevated mental effort is commonly reported by singers, actors, and individuals with voice disorders acquiring a new voice production technique, this phenomenon has yielded almost no empirical investigation. In this study, we captured mental effort associated with intentional voice quality change by using cognitive pupillometry: pupil dilation associated with mental activation- as well as self-report scales of mental effort. Eleven adults (mean age 28, SD 5) were recorded counting out loud in their habitual voice quality as well as three novel qualities: fry, breathy, and twang. Both pupil size and self-report associated with altered voice qualities were significantly larger than for habitual voice production, with medium to large effect sizes. In addition, differences between different target voice qualities were likely due to differences in the physical and emotional demands of producing these voices.

Western diet, characterized by high intakes of saturated fats is a major risk factor for cardiovascular diseases. Plant-based foods, such as nuts, have been shown to be cardioprotective. Baru (Dipteryx alata) is a tree native to the Brazilian Cerrado that produces baru nuts which are rich in monounsaturated fats and antioxidants. This study aimed to examine the effect of baru nut supplementation on blood pressure (BP) and cardiac function. UM-HET3 mice were fed a low-fat, low-sucrose, and -salt (LFLSS) diet alone (n=24) or supplemented with baru (5% w/w; n=12) for four weeks. At week 5, a subset of LFLSS mice were switched to consume a high-fat, high-sucrose and -salt, (HFHSS; n=12) diet while LFLSS + baru was switched to a HFHSS + baru (5% w/w) for an additional 12 weeks. BP was measured biweekly starting at week 4 using tail-cuff plethysmography and cardiac function was assessed using echocardiography at weeks 5 and 15. BP was found to be lower in the baru group after 12 weeks of HFHSS diet compared to the other groups, although not significantly. As expected, 12 weeks of HFHSS diet significantly decreased ejection fraction (EF) and fractional shortening (FS). At the end of the study, baru-supplemented mice had significantly higher EF, FS, and stroke volume (SV) compared to HFHSS alone. Our results suggest that baru supplementation improves cardiac function in mice fed a HFHSS diet. Further studies are ongoing to shed light into the underlying mechanisms.

Chronic inflammation in inflammatory bowel disease (IBD) contributes to tumorigenesis, underscoring the need for targeted delivery systems capable of administering precise, low doses of therapeutics directly to inflamed colonic regions. Inspired by the colon-targeting properties of ginger-derived nanoparticles (GDNPs), we reversely engineered a novel lipid nanoparticle (nLNP) incorporating the three key lipids identified in GDNPs—phosphatidic acid (PA), monogalactosyldiacylglycerol (MGDG), and digalactosyldiacylglycerol (DGDG). Characterization by dynamic light scattering revealed an average nLNP diameter of 190.4 nm with a zeta potential of -22.3 mV. Orally delivered nLNP showed no significant impact on gut microbiota composition and was confirmed as a safe carrier through in vitro and in vivo toxicity tests. Biodistribution studies demonstrated its potential for oral drug delivery, achieving targeted release in colon tissue. Furthermore, fluorescence-activated cell sorting (FACS) revealed that nLNP is internalized by epithelial cells and macrophages, with uptake significantly enhanced in inflamed colon tissues compared to healthy controls. These findings support the development of the nLNP as a safe and effective inflammation-responsive delivery platform for therapeutic interventions in ulcerative colitis.

Eosinophilic esophagitis (EoE) is a chronic inflammatory condition that can negatively affect growth and nutritional status, especially in children. This study evaluates the effects of Dupixent (dupilumab), a biologic therapy approved for EoE, on growth metrics. We monitored the weight and height of 10 EoE patients during Dupixent treatment, comparing them to a control patient diagnosed before the drug's availability, who began treatment in his mid-20s after completing his growth phase and experienced persistent growth stunting despite symptom relief. In contrast, younger patients who started Dupixent demonstrated significant growth spurts, suggesting that early intervention may enhance growth outcomes in EoE patients. These results highlight the critical importance of early diagnosis and treatment of EoE to prevent long-term growth and nutritional issues. The study advocates for a shift in clinical practice towards prioritizing early biologic therapy initiation in pediatric and adolescent EoE patients, potentially leading to improved health outcomes in both symptom management and growth development. Further research is necessary to validate these findings and optimize treatment strategies to address the growth-related challenges associated with EoE.

The global rise of Neisseria gonorrhoeae necessitates a deeper exploration of its biology to address emerging challenges in combating gonorrhea. Optimizing complementation experiments is crucial for understanding the bacterium's complex biology and unraveling genetic factors related to challenges, particularly focusing the requirements for optimum protein translation initiation. Our study investigates the influence of different spacer sequence lengths between the ribosome binding site and start codon on super folder Green Fluorescent Protein (sfGFP) production, serving as a visual readout for protein expression. Colony screening reveals that increasing spacer lengths from 5 to 8 nucleotides leads to a gradual elevation in sfGFP production, while spacers exceeding 8 nucleotides result in decreased production of sfGFP. Ongoing experiments, including fluorescence intensity measurement, flow cytometry, and Western blot techniques, will enable a comprehensive evaluation of protein production across different spacer variants. These findings contribute to a deeper understanding of N. gonorrhoeae molecular biology and offer practical insights for refining complementation and protein production experiments in future studies.

A western diet, characterized by high levels of fats, sugars and salt, is linked to increased levels of inflammation and oxidative stress. The mitogen-activated protein kinase (MAPK) pathway plays a key role in driving inflammation associated with such diets. Blueberries (BB) are well known for their antioxidant and anti-inflammatory properties. Thus, the objective of this study is to assess the mechanisms by which BB exert their cardio-protective effects in mice fed a Western diet. UM-HET3 male mice were either assigned to a control diet (low-fat, low-sucrose, low-salt, LFLSS) alone or supplemented with 5% BB (LFLSS + BB) for four weeks. LFLSS animals remained in the LFLSS or were switched to a high-fat, high-sucrose, high-salt (HFHSS) diet. The LFLSS + BB were switched to consume a HFHSS + BB (5% w/w). Mice were sacrificed and hearts were collected for protein analysis using Western Blot. Data were examined for normality and analyzed using one-way ANOVA or Kruskal-Wallis as appropriate. We observed no changes in the expression of superoxide dismutase (SOD)2, an antioxidant enzyme, and IL-6, a pro-inflammatory cytokine. BB supplementation significantly reduced the phosphorylation of ERK1/2 compared to control (p>0.0316). ERK1/2 is a MAPK that is activated in response to oxidative stress and modulates the expression of IL-6 and SOD2 by activation of transcription factors. This may indicate that BB may reduce oxidative stress thereby downregulating ERK1/2; however, this needs to be confirmed. Further analysis is underway

Respiratory Syncytial Virus (RSV) and human Metapneumovirus (hMPV) are global pathogens and two leading causes of lower respiratory tract infection (LRTI), hospitalizations and deaths in infant, elderly, and immunocompromised populations. Belonging to the Pneumoviridae family, the two viruses share a homologous membrane fusion (F) protein that has shown potential as a target for therapeutic monoclonal antibodies (mAbs). By binding to F proteins at their pre-fusion (preF) conformation rather than post-fusion (postF) conformation, mAbs can neutralize viruses and block their entry into host cells. Nanobodies are antigen-binding entities derived from the heavy chain-only antibodies of camelid animals, and they retain full antigen specificity at a very small size, creating unique potential advantages over traditional approaches. Here we sequentially immunized llama with prefusion stabilized hMPV and RSV F proteins, and identified two nanobodies, NB16 and NB28, that target a conserved epitope on preF proteins. NB16 and NB28 showed strong binding affinity with both viral preF proteins and demonstrated cross-neutralization capabilities against the viruses. To further increase neutralization efficacy, in vitro maturation was performed by site directed mutagenesis in the variable regions of the nanobody sequences, and all cross-binding mutants were isolated for comparisons with the parent sequence. Further binding analyses and neutralization assays will be performed with the goal of identifying nanobodies with increased affinity for preF proteins and high levels of viral neutralization. Our work provides a framework of developing countermeasures against LRTI caused by HMPV and RSV and establishes a generalizable strategy of nanobody-based therapies development against other viral pathogens.

A high-fat diet increases oxidative stress and immune dysregulation in the gut, which negatively affects tight junction protein expression. Blueberries (BB) have antioxidant properties and research shows BB consumption strengthens the intestinal barrier. This study aims to determine the effects of BB supplementation on the intestinal health of mice fed a high-fat, high-sucrose, high-salt (HFHSS) diet. Four-week-old UM-HET3 male mice were fed a control diet (low-fat, low-sucrose, low-salt, LFLSS) alone or supplemented with BB for four weeks. Mice consuming LFLSS remained on a LFLSS diet or were switched to a HFHSS. Mice on LFLSS + BB were assigned to HFHSS + 5% BB for 12 weeks. Colons were excised and expression of pro-oxidant and antioxidant enzymes were examined via Western Blot. Data were analyzed using one-way ANOVA followed by Tukey’s posthoc test. Protein expression glutathione peroxidase (GPx)-1 was significantly downregulated by HFHSS (p = 0.007), and BB was unable to attenuate this effect (p=0.3). Though not significant, the expression of GPx-3 was also down-regulated by HFHSS diet, which was prevented by BB supplementation (p = 0.01). Lastly, the expression of heme oxygenase (HO)-1 and NADPH oxidase (NOX)-1 were not different across groups. Results indicate that BB supplementation does not mitigate the detrimental effects of HFHSS on the expression of antioxidant markers in the gut. Further analyses are ongoing to examine its impact on intestinal inflammatory markers and tight junction proteins.

Objective: High-fat, high-sugar, and high-salt (HFHSS) diets contribute to cognitive decline and neuroinflammation, highlighting the needs for dietary interventions to support brain health. Blueberries (BB), rich in anthocyanins, have shown promise in improving cognitive function and reducing oxidative stress. This study examines the effects of BB supplementation on cognitive function and neuroprotection in mice fed a HFHSS diet. Methods: UM-HET3 male mice were fed a low-fat, low-sugar, low-salt (LFLSS) diet with or without BB (5% w/w) for four weeks. Mice were then switched to a HFHSS diet (n=9) or remained in the LFLSS diet while LFLSS + BB were switched to a HFHSS + BB (n=9) for 12 weeks. Behavioral tests included Novel Object Recognition (NOR) at week 9, Barnes Maze at weeks 10–12, and Fear Conditioning at weeks 14–15. Results: HFHSS + BB group showed no significant differences in freezing time, freezing episodes, or latency compared to the other groups (p>0.05). However, a trend toward fear extinction was observed in the BB group, indicated by shorter freezing episodes (p>0.05). Conclusion: While BB supplementation did not significantly alter behavior in the HFHSS diet model, it may have a modest impact on fear extinction. NOR and Barnes Maze data analyses is underway as is hippocampal protein expression analyses for oxidative stress and inflammatory markers. Keywords: Blueberry supplementation, cognitive function, anthocyanins, UM-HET3 mice, high-fat diet, BDNF.