Poster Session C

In this work a NIR emitting dye, p-Toluenesulfonate (IR-813) was explored as a model precursor to develop red emissive carbon dots (R-CD) for fluorescent imaging.  Current commercial photosensitizers (PS) used in fluorescence-based techniques have glaring limitations.  For example, to achieve optimal efficiency, PS used in theranostics techniques require the utilization of light within the red region to NIR-I window ranging between 650-950 nm, respectively.  A direct consequence of this requirement typically results in the degradation of the PS such as photobleaching or thermal degradation.  Additionally, modern PS have the disadvantage of non-rapid blood clearance from the host which limits biomedical applications.  Significantly, (CDs) have attracted increasing attention in various biomedical areas of application such as in vitro/in vivo imaging, photodynamic and photothermal therapy.  Previous literature suggests that complex surface modifications of produced CDs with hydrophobic organic dyes as the precursor are of interest to unlock the potential route to develop red emissive CDs.  If one were to develop carbon dots (CDs) from this source, it is expected that the product will retain most of the functionality of the precursor.  In this case, mainly utilization in the far-red region between 700 – 900 nm respectively.

The stress of a cancer diagnosis has adverse effects on the family caregiver's daily diet and subsequent immune health. Such relation may be exacerbated by limited resources, such as income.  Less known is the extent to which daily diet and income relate to immune health of family caregivers of adult cancer patients. This study investigates the association of diet with inflammatory biomarkers and the moderating effects of poverty among cancer caregivers concurrently and prospectively.
Participants reported consuming at least 5 servings of fruits and vegetables 3 days a week and fat 4-5 times a week. One-fifth of the sample met the criterion for poverty. IL-6 and IL-10 levels at T1 and T2 were comparable with those reported in other caregiver studies, and changes in levels from T1 to T2 were not significant. Hierarchical regression analyses revealed that higher FatC at T1 was associated with lower IL-10 at T1 (B = -.149, p = .017) and increased IL-6 from T1 to T2 (B = .302, p = .026).  Higher FVC at T1 was associated with increased IL-10 from T1 to T2 (B = .146, p < .001). These diet effects were independent of those of poverty level of income.
Findings indicate that dietary habits, regardless of income, uniquely contributes to caregiver's inflammatory biomarkers. Evidence suggests that high fat diets have both concurrent and prospective deleterious effects on caregiver's immune health, whereas diets consisting of high fruits and vegetables have lasting protective effects. 

Background: Diffuse large B cell lymphoma (DLBCL) is the most common lymphoma diagnosis. Targeted therapeutics originally focused on the cell-of-origin hypothesis, which divides DLBCL into subtypes based on the origin of oncogenic cells. However, newer classifications point to a wider diversity of subtypes and one gene that is mutated in approximately 5% of newly diagnosed DLBCL encodes the BCL10 protein. BCL10 associates with CARD11 and MALT1 to activate downstream oncogenic pathways such as the canonical NF-kB and JNK. The mechanisms by which BCL10 mutations promote lymphomagenesis and drug resistance are poorly understood. We hypothesize that BCL10 mutations promote oncogenesis and contribute to drug resistance through activation of NF-kB signaling. Methods: We used a luciferase dual reporter assay to probe NF-kB activity in 293T cells with wild-type or mutant BCL10. Overexpression of two recurrent mutations was performed using a doxycycline-inducible system in DLBCL cell lines and Western Blot was used to identify changes in signaling pathways. Next, we used a human phosphokinase array to determine the profile of phosphorylated proteins and RNA sequencing to understand changes in gene expression. Results:Our data indicate upregulation of both canonical and noncanonical NF-kB pathways and increased MALT1 activity. Additionally, the phosphokinase array and RNA sequencing data showed upregulated ERK signaling and activation of cytokine-mediated signaling. Conclusion: Our results demonstrate mechanisms through which BCL10 mutations may promote lymphomagenesis and identify potential drug targets. Further analysis of targetable signaling pathways, including ERK and cytokine signaling, are underway.

Fluorescence recovery after photobleaching (FRAP) is a technique that uses a confocal microscope to study the diffusion of molecules within a membrane. During FRAP, fluorescently-labeled molecules within a specified region of interest (ROI) are photobleached using a high-power laser. After photobleaching, the fluorescent intensity in the ROI is reduced to zero. Due to molecular diffusion that naturally occurs within the membrane, surrounding fluorescent molecules will move into the bleached area. The confocal microscope is used to visualize the recovery of fluorescent intensity within the ROI over time. Analysis of the recovery curve must be performed to extract meaningful quantitative data, such as the diffusion coefficient and the halftime recovery of the sample. Having insight on these properties is crucial for understanding molecular diffusion within a given material. Commercial confocal microscopy software can perform FRAP, yet it lacks the analytical tools to extract these properties. Therefore, in this project, a user-friendly interface was developed in MATLAB to curve fit the data, extract the diffusion coefficient and halftime recovery, and visualize the confocal images. Proof of concept testing using FRAP data of insulin diffusion through a polymer capsule demonstrated the efficacy of the interface.

Cognitive flexibility is the ability to switch between a set of task rules, while affective flexibility is the ability to switch between emotional and non-emotional aspects of a stimulus.  Although there is research examining both of these types of flexible judgements, few studies have directly compared them.  The present study aims to compare cognitive flexibility judgements (SwitchNumbers) with affective flexibility judgements (SwitchImages) utilizing reaction time (ms) and functional magnetic resonance imaging (fMRI).  Behavioral data (n=20) and neuroimaging data (n=19, due to data loss) were collected.  Stimuli were presented via E-prime 3.0 software with eight task conditions: 4 non-switch, and 4 switch blocks. The contrast of conditions SwitchImages - SwitchNumbers showed activation within the central executive network, with some activation within the limbic system. Additionally, the SwitchImages condition had the longest reaction time among all conditions. These results suggest that switching between emotional and non-emotional aspects of stimuli is more difficult than single task conditions and cognitive flexibility conditions, likely due to the increased cognitive load.

Cancer patients are vulnerable to neuroendocrine dysregulation, implicating cancer progression and poorer health outcomes. Resilience through spirituality may promote better adjustment to cancer diagnosis. This study examined the degree to which patients' spirituality is associated with neuroendocrine biomarkers of colorectal cancer patients and the moderating role of Hispanic ethnicity. Domains of spirituality (FACIT-Sp: meaning, faith, and peace) and ethnicity (Hispanic vs. non-Hispanic) were self-reported. Cortisol, alpha amylase (sAA), and DHEA-s were assayed from saliva samples collected at waking and bedtime on seven consecutive days. Mean levels and diurnal slopes over seven days were calculated. Age and cancer stage were covariates. Patients reported moderate to high levels of meaning, faith, and peace, and displayed lower levels of cortisol, sAA, and DHEA-s and blunted diurnal slopes over a day. General linear modeling revealed that greater faith was associated with lower levels of cortisol and DHEA-s at waking (|B| ≥ -0.477, p ≤ .042). Greater peace was associated with steeper sAA diurnal slope (B = 0.021, p = .005). Hispanic patients with greater faith showed lower levels of sAA at waking and at bedtime (B ≤ -125.305, p ≤ .018), and steeper sAA diurnal slope (B = 0.028, p = .022), which was not seen among non-Hispanic patients. Findings highlight that making peace with a cancer diagnosis and drawing on one's faith, particularly among Hispanic patients, protects against neuroendocrine dysregulation. Spirituality-based interventions may facilitate health-promoting adjustments of Hispanics during the early cancer survivorship phase. 

The lack of access to accurate and comprehensive sexual health education in the United States continues to be a pressing public health issue. The Sex Wrap (TSW) is a multimedia sexual health podcast and social media presence that educates its followers on sexual health and ancillary themes including sexuality, relationships, and communication. TSW's Instagram account revamps the evidence-based sexual health information from the podcast as original or reposted content. With internet memes becoming a driving cultural force among young adults aged 14-29, the podcast utilizes its Instagram social media presence to effectively stage online health-based social marketing. Comedic value aside, the highly accessible nature of memes makes them a powerful tool for information dissemination. The meme-based sexual health intervention via TSW's Instagram increased engagement, improved reach, and converted followers to podcast listeners. The insight data also demonstrated how employing social media can be used to address disparities in sexual health education caused by the failure of the abstinence-only-until-marriage curricula. This case study demonstrates how social-media based interventions can be used to fill social and legislative gaps in sex education in a comprehensive way.

The RAS oncogene, which is mutated in over 95% of pancreatic cancers, relies on reactive oxygen species (ROS) to invoke oncogenic signaling that allows tumor cells to proliferate and survive. However excess ROS can also kill or proliferatively arrest cancer cells through oxidative stress and DNA damage. Thus RAS-driven cancer cells elevate redox-protective adaptations to enable ROS-driven signaling but prevent ROS-driven tumor inhibition. The mammalian Nudix pyrophosphatase MTH1(MutT Homolog 1), which eliminates oxidized nucleotides to prevent their incorporation as oxidative damage into the genome, is a critical redox-protective adaption in RAS-driven tumor cells. Our lab has previously shown that depletion of MTH1 alters the ability of RAS-driven cancer cells to drive ROS-driven signaling. In pancreatic cancer, a major ROS oncogenic signaling hub is the epidermal growth factor receptor (EGFR) pathway, which regulates factors such as growth, proliferation, and differentiation through RAS. We hypothesized reducing redox protection from MTH1 would compromise this pathway. We found that depleting MTH1 using small hairpin RNAs (shRNA) in pancreatic cancer cell lines reduces their proliferative ability. This inhibition is associated with a decrease in EGFR expression. Interestingly analysis of PDAC (pancreatic ductal adenocarcinoma) patient datasets showed MTH1 and EGFR levels are positively correlated. Thus MTH1 inhibition could supplement or even replace EGFR inhibitors as treatment possibilities in pancreatic cancer, which has few durable treatment options and is fatal.  

Coral restoration efforts aim to preserve biodiversity and build self-sustaining, sexually reproducing coral populations. It is important to increase coral growth and survivorship, genotypic diversity, and enhance overall resilience. Historically, coral practitioners exclusively utilized corals from their own regions. Leveraging novel assisted migration, outplanting, and tracking methodologies, the largest data set of coral survivorship and productivity along Florida's Coral Reef was established and showed that it is possible for non-native genotypes to survive and thrive in new regions. In June 2020, five nursery programs, Nova Southeastern University, the University of Miami, Coral Restoration Foundation, Fish and Wildlife Commission, and MOTE Marine Laboratory, participated in the first region-wide coral genotype swap. This involved the exchange of >90 genotypes of Acropora cervicornis originating from dozens of reef locations between Broward County and the Florida Keys. While growth rates, measured 6-7 months later varied significantly, the study proved that these corals can survive large-scale relocation and thrive in varying environments. The data collected in this unprecedented large-scale assisted migration study can be incorporated into future regional restoration frameworks.

Organoids are 3D cellular structures for organ modeling and drug testing. Retinal organoids are needed to study diseases like retinoblastoma; however, their development is particularly difficult. To form inner retina cells, the organoids need to be in a hypoxic state, yet the outer retina cells consume more oxygen than any other tissue in the body. It is possible to simulate this environment with a bioreactor by utilizing an open well design for ease of use. To accommodate the culture media consumption without disturbing the organoids in the open wells, it requires an external microfluidic system. A KD Scientific syringe pump with four 30 mL syringes simultaneously infuses and withdraws 600 nL of media per minute through 1/32 inch autoclavable Tygon tubing. The tubing is connected to the bioreactor through an innovative housing device that uses Stereolithography 3D printing technology with a specialized resin. Unlike most 3D printed plastics, the resin is autoclavable for sterilization and incubation. The tubing can be seamlessly and quickly removed and replaced to monitor the bioreactor without disturbance.Simulations using COMSOL show that the oxygen gradient is not affected by the fluid flow, which is confirmed by our measurements using an oxygen sensor. Implementation of the system resulted in successful organoid survival over a period of multiple weeks. Although testing is required to confirm the bioreactor can be imaged within the container, the system is a promising option for providing culture media to an open well organoid bioreactors over long periods of time.

Disease modelling is an extremely powerful tool that provides insight to disease pathology in areas of the body that cannot be easily studied. 3D Organoid culture has become a hallmark in disease modelling, especially for specialized tissues of the brain which are impossible to study at the cellular level in an afflicted individual. There is a need to understand the disease mechanisms of genetic-based inner ear hearing loss, and while organoid culture shows great promise, there is one critical limitation. The part of the inner ear most affected by the genetic variation causing deafness is the cochlea, and during development the portion of the brain destined to become the cochlea is exposed to a gradient of growth factors which will promote the specialization of the tissue. Generating a stable concentration gradient of growth factors over long periods of time is extremely difficult using traditional cell culture methods, and therefore, the resulting organoids do not exhibit this specialization that is seen during cochlear development. The proposed Chip design utilizes laminar flow and simple diffusion to generate a concentration gradient. Stock solutions continuously flowed into the chip generates a difference in concentration via laminar flow while simple diffusion occurs to generate a concentration gradient that extends from the region of high growth factor concentration to the region of low growth factor concentration. An organoid exposed to this concentration gradient will undergo specialization similar to that seen during cochlear development, making it a more accurate disease model of genetic-based hearing loss.

A community's built environment plays a tremendous role in the well-being of its members. Access to grocery stores, affordable healthy food options, or a surplus in cheap fast food can significantly affect the health of the individuals living in that area. The purpose of this study is to examine the relationship between food deserts and the health of communities of color in Miami-Dade county. We used data from existing databases, MiamiDadeMatters.org and the United States Department of Agriculture food map, to gather the demographics, food insecurity index, and obesity rates of each zip code in Miami-dade county. We ran statistical tests to measure the relationships between our variables and found that communities of color are disproportionately affected by food deserts compared to predominantly white communities. As zip codes' food insecurity increased so did obesity rates. Zipcodes with high percentages of black residents had a significant positive correlation with obesity rate, whereas zip codes with high percentages of white populations had a significant negative correlation with obesity rates. These findings reflect the significant systemic disadvantage communities of color experience regarding food access and their health. This study can be used to address this disparity in policymaking and community advocacy initiatives.

While gastrulation is unique to metazoans, the site at which it occurs is not uniform for all metazoans. Non-bilaterians and bilaterians diverged from an urbilaterian common ancestor, and with that divergence came a change in the site of gastrulation. In observed non-bilaterian taxa, such as Cnidaria and Ctenophora, endoderm specification and archenteron formation occur at the animal pole, allowing for gastrulation to occur here. On the other hand, bilaterian taxa experience this process at the vegetal pole. It is not only the difference in the site of gastrulation that is of interest, but it is that the site of gastrulation is completely opposite in polarity. Bilaterian organisms have evolved to develop extremely complex functions, and it is believed that the switch in polarity from the divergence from non-bilaterians may have an important role in their complex development. The cnidarian, Nematostella vectensis, has been shown to utilize the Wnt/Planar Cell Polarity pathway protein Strabismus for bottle cell formation and initial gut invagination, which is localized to the animal pole and allows for gastrulation to occur. This protein is of interest to determine if it possesses a function in the gastrulation of bilaterian organisms. It is hypothesized that if Strabismus is present in bilaterian taxa, it will be localized to the vegetal pole and function in the initial invagination for gastrulation to occur. Determining the molecular mechanism for gastrulation in both non-bilaterian and bilaterian taxa is important in understanding the evolution of the polarity of metazoan embryos.

There has been a significant decrease in American's support for the death penalty over the years. Consequently, we have witnessed more states abolishing the use of the death penalty. According to the data from the General Social Survey (GSS) 57% of Americans aged 65+ were in favor of the death penalty. This study analyzes the attitudes of the undergraduate population at the University of Miami towards the death penalty to observe general trends in attitudes among a younger sample. In this sample (N=200) with a mean age of 20 years old, only 32.5% of students favored the death penalty. A cross tabulation was performed to further analyze students' attitudes towards the use of the death penalty based on the following variables: major, gender, year group, race, and political affiliation. Of all the variables studied, only political affiliation was found to have a significant difference in attitudes. The chi square was 31.956, (x2 =31.956, p = <0.001). From this information we then looked at how each political affiliation differed based on attitudes toward the death penalty for different types of crimes using an ANOVA test. Of all the types of crimes studied, the following were found to be significant: 1st degree murder (F3,194 = 9.840, p= <.001), 2nd degree murder (F3,194 = 4.668, p= .004), 3rd degree murder, (F3,191 = 3.849, p= .010), rape (F3,192 = 2.756, p= .044), robbery (F3,191 = 2.815, p= .040), aggravated assault (F3,191 = 3.750, p= .012), arson (F3,191 = 7.044, p= <.001). 

Pancreatic islet microencapsulation and localized delivery of immunomodulatory molecules may improve islet transplantation outcomes in type-1 diabetes (T1D) patients by preventing allograft rejection and maintaining metabolic control without chronic and systemic immunosuppression. Here, we encapsulated human islets (HIs) in poly(ethylene glycol) conformal coating (CC), by mixing HI with minimally crosslinked 8a-PEG-maleimide and 36.2% (w/v) HS-PEG-SH, extruded through a custom fluidic device (Biorep) using a PPG+10%Span80 external oil solution and a 1.65mg/ml DTT/PPG gelling emulsion.  We measured in vitro functionality through dynamic GSIS and insulin ELISA (Mercodia), and in vivo by glucose monitoring of diabetic NOD-scid mice transplanted with 2000 IEQ of CC-His in their fat pad. Biocompatibility was assessed after tissue explantation through histological and immunofluorescence evaluation using DAPI and mac2 antibodies. Micelles and nanofibers nanoparticles were made of PEG-poly(propylene sulfide) and PEG-oligo(ethylene sulfide) di-block copolymers, respectively. We tested 5 and 10 µg/mL Dex-nanoparticles inhibition of Raw264.7 macrophage cytokine production after lipopolysaccharide stimulation (ELISA), and studied whether 1mg/Kg Dex-nanoparticles improve CC biocompatibility in vivo 21 days post CC transplantation. We found that CC HIs maintain physiological insulin secretion in vitro and in vivo. However, their biocompatibility in vivo is decreased by the presence of macrophages. We demonstrated that Dexamethasone micelles and nanofibers are efficient in decreasing inflammatory cytokines in vitro, however mac-2+ cells are around CC capsules even if treatment is present.

Bacteria in humans are becoming increasingly important to modern medicine. They are understood to play roles in hundreds of human systems from mood regulation to metastatic colonization in breast cancer. They even decide what foods you will crave. This study takes a bidirectional approach to understanding the gut microbiome through characterization of a Whole Cell Biosensor (WCB) capable of detecting the presence and quantity of novel quorum sensing molecule 3,5-dimethyl-pyrazine-2-ol (DPO), as well as a biogeographical approach to the large intestine microbiome through 16S Fluorescent In-Situ Hybridization (FISH). We were able to effectively characterize the biosensor for future use in-vivo, as well as design a working protocol for the 16S FISH technique to be used in targeting other microbial mechanisms in the mouse large intestine. This presents a valuable approach to understanding the complex interactions of the gut microbiota, as well as a potential method for early detection of virulence in-vivo.

The project's overall objective is to study afferent nociceptor activity in the corneas of transgenic
mice as a novel method for studying oral cancer pain. Using a scanning laser confocal for Ca2+
imaging in vivo, we will assess the change in fluorescence of Ca-reporter GCaMP3 expressing
sensory neurons in response to a known algesic stimulus (10-100 μM capsaicin). This activity
will be compared to that from supernatants of cultures of oral cancer cells. The supernatant will
be obtained from our colleague, Dr. Brian Schmidt. The experiments will also involve the use of
a corneal recording chamber (engineered by our mentor, Dr. Stephen Roper) and a
microperfusion "picospritzer" system. Many issues still exist in terms of optimal protocol. One
obstacle is to stabilize the mouse during experiments without changing the neuron's response to
capsaicin. Nerve fibers in the eye must remain stable, within a few microns, over multiple trials
in order to be able to analyze the data. Another issue is how to control the currents of the tyrode
bath around the eye that impacts capsaicin perfusion. There is an inflow and outflow system
that washes away the capsaicin, causing turbulence even within the short micron distance
between the picospritzer and the cornea. This research will help to develop optimal dosage and
imaging protocols for studying corneal nociceptor activity and analyzing neural responses to
capsaicin.

Studying all the supraspinal projecting populations after spinal cord injury has been difficult because sectioning of tissue and analysis of 2D images of a whole brain is time consuming and laborious. We developed methods that can image the whole mouse brain without sectioning using tissue clearing methods and light sheet microscopy. The present study investigates, in more detail, most of the projecting neurons and mainly quantification using enhanced 3D imagining of most areas including those that been historically overlooked in the spinal cord injury field. This study incorporates newer versions of fluorescent proteins such, as mScarlet and mGreenLanternallow for greater and more precise detection in critical supraspinal areas such as the brainstem. Using 3D software such as Imaris allows us for more accurate, uniform, and overall accessible quantification and labelling of these areas. These advancements in detection and examination allow for contribution and generation of databases. This is especially important regarding closing the gap in dissimilarities and inconsistencies in literature and provide and educational resource for those lacking the expert knowledge for studying these brain areas. Furthermore, this type of approach will provide essential information and tools for those studying regenerative therapies.