THURSDAY, APRIL 27, 2023
8:00 AM UNTIL 11:00 AM
ABSTRACTS
Postdoctoral Fellows
GRADUATE STUDENTS
PHARM.D. STUDENTS
UNDERGRADUATE STUDENTS
Cerebral palsy (CP) is the most common form of physical disability. CP arises from a variety of perinatal insults and results in permanent, lifelong motor deficits, most often spasticity (Rosenbaum et al., 2007). Spastic CP is thought to arise from damage to the cortex and corticospinal tracts (CSTs) which results in downstream disinhibition of spinal motoneurons (MNs) and hyperreflexia, although the specific mechanism has never been directly tested (Oskoui et al., 2013, Volpe, 2017).
Perinatal hypoxia-ischemia (HI) is a factor in many CP-causing injuries. Rabbits subjected to prenatal HI in late gestation show motor deficits similar to spastic CP including hypertonia and hyperreflexia (Derrick et al., 2004, Synowiec et al., 2019), thus allowing a deeper investigation into mechanisms of spasticity. In the spinal cord, peptidergic nociceptors and CST projections converge directly onto GAD2+ GABAergic interneurons (GABApre) that mediate primary afferent depolarization (PAD), however changes in spinal circuits after perinatal insults are poorly understood.
We hypothesize that loss of CST projections in CP allows expansion of nociceptive innervation of GABApre neurons, increasing conduction of Ia afferent spikes causing reflex irradiation.
We performed extracellular recordings in the in vitro isolated spinal cord of control and HI neonatal rabbits (P 1-5). Stimulation of ventral or dorsal roots were performed to evoked monosynaptic reflex (MSR) and dorsal root potentials (DRPs) respectively. We observed a larger spread of the MSR and DRPs in the HI rabbits compared to control rabbits, suggesting physiopathological changes in spinal circuits in the HI kits. Pharmacological dissection of DRPs (an indirect way to measure PAD) indicates that they origin of this is GABAergic. We can conclude that
1) PAD is GABAergic.
2) PAD circuit is augmented HI rabbits (larger DRPs).
3) Reflex irradiation is increased in HI rabbits (MSR). All of this shows that in the HI rabbits spinal neuronal circuits and sensory afferents are dysfunctional (not just the brain!).
Cerebral palsy (CP) is caused by a variety of factors that damage the developing central nervous system. Impaired motor control, including muscle stiffness and spasticity, is the hallmark of spastic CP. Rabbits that experience hypoxic-ischemic (HI) injury in utero (at 70-80% gestation) are born with muscle stiffness, hyperreflexia, and, as recently discovered, increased serotonin (5HT) in the spinal cord. To determine whether serotonergic modulation of spinal motoneurons (MNs) contributes to motor deficits, we performed ex vivo whole cell patch clamp in neonatal rabbit spinal cord slices at postnatal day (P) 0-5. HI MNs responded to application of α-methyl 5HT (a 5-HT1/5-HT2 receptor agonist) and citalopram (a selective 5-HT reuptake inhibitor) with hyperpolarization of persistent inward currents and threshold voltage for action potentials, reduced maximum firing rate, and an altered pattern of spike frequency adaptation while control MNs did not exhibit any of these responses. To further explore the differential sensitivity of MNs to 5-HT, we performed immunohistochemistry for inhibitory 5-HT1A receptors in lumbar spinal MNs at P5. Fewer HI MNs expressed the 5-HT1A receptor compared to age-matched controls. This suggests many HI MNs lack a normal mechanism of central fatigue mediated by 5- HT1A receptors. Other 5-HT receptors (including 5-HT2) are likely responsible for the robust increase in HI MN excitability. In summary, by directly exciting MNs, the increased concentration of spinal 5-HT in HI rabbits can cause MN hyperexcitability, muscle stiffness, and spasticity characteristic of CP. Therapeutic strategies that target serotonergic neuromodulation may be beneficial to individuals with CP.
The maintenance of specific cell morphology and function within a tissue relies on the ability of cells to respond and adapt to dynamic changes in their micro-environment. This response consists in bidirectional feedback between microenvironmental signals, the contractile cytoskeleton that responds to these signals and determines cell shape, and the transcriptional circuitry that establishes and maintains cell identity. Extracellular forces exerted from the extracellular matrix (ECM) to the plasma membrane via adhesion molecules are transmitted to the cytoskeleton and then to the nucleus, ultimately resulting in changes to nuclear morphology, chromatin compaction/organization, and gene expression to adapt cell features to the environment mechanics. During aging occurs the accumulation of intra-intermolecular covalent bonds (crosslinks) between molecules with slow turnovers, such as collagen and elastin of the ECM. These crosslinks affect the physical properties of the ECM (i. e. stiffness) forcing cells to adapt to the changing environment. These physiologic changes of brain mechanical properties make the mechanical response of neurons crucial for the maintenance of their function and their survival. In the present study we show the effect of substrates (Matrigel and PDL/laminin) with different mechanical features on C9orf72 iNs (C9exp iNs) and isogenic controls (C9iso iNs). Our data demonstrate an alteration of the mechanical adaptation of C9exp iNs to high stiffness substrates affecting cell morphology and gene expression. Furthermore, we found the alteration of this complex pathway involves many different steps of the mechanical signaling cascade from the focal adhesions to the nucleus.
Alcohol-related liver disease (ALD) is the most prevalent form of liver disease commonly associated with hepatocellular carcinoma. The current anti-inflammatory, anti-viral, and immunosuppressive therapies for liver inflammation are not consistently effective and safe. Therefore, effective and timely interventions for targeted and sustained treatment of chronic ALD remain an urgent and unmet clinical need. The G-protein coupled bile acid receptor TGR5 (Gpbar1) expressed in Kupffer cells (KCs) in liver has a potent anti-inflammatory effect. Also, we incorporated INT 777, a highly selective and potent semisynthetic bile acid agonist of Gpbar1 attenuates pro-inflammatory cytokine production by KCs, into carboxymethyl chitosan (CMC) coated poly lactic co glycolic acid (PLGA) nanoparticle (NPs) to target KCs. In addition, CMC, a pH-responsive biodegradable polymer, was used to trigger the release of the anti-inflammatory drug dexamethasone (DEX) in response to the acidic pH in the inflamed liver microenvironment. Therefore, concurrent NP-mediated Gpbar1 stimulation and pH-responsive DEX release in the acidic microenvironment have an additive effect in mitigating chronic hepatic inflammation. The NPs were developed using acetone and dichloromethane (1:1 ratio) as a solvent using a single emulsion technique in a 5% (v/v) polyvinyl alcohol (PVA) solution. INT 777 was conjugated to NPs using a carbodiimide coupling reaction. The synthesized NPs showed good particle size (<300 nm), zeta potential, and stability. The INT conjugation was confirmed using x-ray photoelectron spectroscopy (XPS), and cytocompatibility studies confirmed that NPs were biocompatible. The DEX release study confirmed the pH-responsive release of DEX from NPs. The cell uptake study using THP1, a leukemia monocytic cell line representing KCs, showed that INT-conjugated particles targeted THP1 cells. The ALD was developed in a mouse model by giving a 5% (v/v) ethanol diet and injecting NPs with INT. The results confirmed the inflamed liver and accumulation of NPs in the liver.
Background/Objectives: As of April 2023, 22 states and jurisdictions have a protocol in place to allow for autonomous prescribing of hormonal contraceptives by pharmacists. The American College of Obstetricians and Gynecologists (ACOG) has expressed their support for pharmacists prescribing hormonal contraceptives in addition to over-the-counter access to mitigate barriers to access. The objective of this study was to simultaneously assess student, pharmacist, and other clinician perspectives on pharmacist-prescribed hormonal contraceptives, to support expanded pharmacist authority to prescribe hormonal contraceptives in Rhode Island.
Methods: Researchers conducted an observational study via a cross-sectional survey distributed to students, licensed pharmacists and clinicians. This study was IRB approved through the University of Rhode Island as exempt and data were collected from October to December 2022. The survey required approximately 8 minutes to complete and assessed feelings and perspectives towards pharmacist prescribed hormonal contraception, likelihood of receiving pharmacist prescribed hormonal contraception and pharmacist comfort with prescribing hormonal contraceptives.
Results: Nearly 90% of students, 80% of pharmacists and 70% of clinicians either agree or strongly agree with pharmacist prescribed hormonal contraception. Sixty-one percent of pharmacists said that they are either very or somewhat comfortable prescribing hormonal contraception. The most frequently reported barriers in accessing contraception for all respondents, were time/delay in appointment, cost/copay, and lack of gynecologist. Approximately 76% of students said they were extremely or somewhat likely to obtain a hormonal contraception prescription from their pharmacist.
Implications/Conclusions: The majority of pharmacists, clinicians, students and in Rhode Island support pharmacist prescribed hormonal contraceptives and the most frequently reported barriers faced in accessing contraception can be overcome through pharmacist prescribing. The information and data gathered from this study has already had an impact to support pending legislation and implementation of prescriptive authority and expanded access to contraception in the State of Rhode Island.
Fibrillar amyloid β-protein (Aβ) deposits in brain, which are primarily composed of Aβ40 or Aβ42 peptides, are key pathological features of Alzheimer’s disease and related disorders. Structural studies on mature A 40 fibrils formed in solution have suggested they adopt a two stranded (U-shaped) structure whereas studies on A 42 fibrils typically show a three-stranded structure. Fibrillar amyloid deposits in brain promote activation of astrocytes and microglia although the underlying mechanisms are still not clear. Here, we treated primary astrocyte and microglia cells with Aβ40 or Aβ42 fibrils, and bulk RNA sequencing was performed. According to the Pearson’s correlation result, astrocytes and microglia cells undergo a great number of Aβ peptides-induced changes in gene expression and Aβ42 and Aβ40 peptides showed similar effects on glia cells compared with the control group. To elucidate the gene changes after Aβ peptides treatment, we conducted the differential gene expression analysis to compare Aβ42 and Aβ40 glia cells. The expression data were set with limits of ≥5-fold increase in expression and p < 0.05. A greater number of genes were differentially expressed by Aβ42 treated glia cells compared with the Aβ40 treated glia cells. Immunohistochemical analysis validated our RNAseq data by performing in an AD rat model with parenchymal fibrillar Aβ42 deposits confirming the expression of MMP9, MMP12, PAI-1 and C1r in plaque-associated microglia and iNOS, GBP2 and C3D in plaque-associated astrocytes. Ingenuity Pathway Analysis further highlighted that Aβ42 treatment up-regulated cellular activation pathways and immune response pathways in glia cells. Interestingly, we found that astrocytes treated with Aβ fibrils up-regulated a larger number of A1 reactive genes compared with A2 reactive genes, and microglia treated with Aβ fibrils upregulated more M1 state genes than M2 state genes. In summary, ex vivo studies show that the distinct structures of Aβ40 and Aβ42 fibrils differentially stimulate the gene expression profile of glia cells and that Aβ42 fibrils are more potent stimulators, together with the intracerebral injection with Aβ fibrils, we are aiming to provide a detailed insight in the pathogenesis of AD and related amyloid-depositing disorders.
Ubiquitin specific peptidase 2 (USP2) is a deubiquitinating enzyme known to modulate cell cycle progression, circadian rhythm, and carcinogenesis in the liver. USP2 and its isoforms USP2a and USP2b are dysregulated in human hepatocellular carcinoma (HCC) tissues and farnesoid x receptor knockout (FXR-KO)-HCC mouse models. However, the mechanism by which FXR regulates USP2 isoforms and its contribution to the pathogenesis of HCC is largely unknown. In this study, we discovered that USP2a and USP2b are derived from separate promoters and are specifically regulated by ligand-activated FXR. Our in vivo studies revealed that USP2a and USP2b expression is significantly increased in obetocholic acid (OCA)-treated wild-type (WT) mice and have no change in FXR-KO mice compared to vehicle treated groups. In our in vitro studies, we discovered that the FXR isoform FXRα2 predominantly transactivated USP2a and USP2b promoter activity in Huh7 cells. In silico analysis of USP2a and USP2b promoters in bioinformatic predictions revealed novel USP2a and USP2b-FXR response elements (FXREs). Targeted mutagenesis of the novel FXREs completely abolished inducibility of the USP2a and USP2b promoters in FXR agonist OCA-treated Huh7 cells. Electrophoretic mobility shift assays (EMSA) and chromatin immunoprecipitation (ChIP) demonstrated direct binding and abundant recruitment of FXR and its coregulators to the human and mouse USP2a and USP2b promoters. Overall, this study has established the correlation between the expression of USP2 isoforms and their direct regulation by FXR in the pathogenesis of HCC. Most importantly, this novel mechanism provides the molecular basis for developing therapies to treat HCC through modulating USP2a and USP2b expression by FXR activation.
Recent decades have witnessed a dramatic increase in human longevity, which has contributed to higher prevalence of age-related diseases including brain aging disorders, such as Alzheimer’s disease (AD). The aging process is accompanied by an accumulation of damage to macromolecules, organelles, and cells, which ultimately leads to organ/tissue dysfunction and death. Although the precise cause of the aging process is unknown, epigenetic alterations and deregulation of gene expression have been implicated in playing a role. Using the innovative ICE (inducible changes to the epigenome) mouse model together with the well-characterized APP/PSEN1 (APP/PS1) mouse, we are directly testing, for the first time, whether epigenetic alterations induced by DNA damage, can affect the onset and progression of AD pathology in “DICE” (dementia from inducible changes to the epigenome) mice. A battery of behavioral testing is ongoing to compare possible cognitive changes in DICE mice with APP/PS1/CRE, ICE, and CRE controls. Preliminary results thus far indicate that male and female DICE mice move significantly more and with faster speed than controls, when assessing spontaneous locomotion in the open field behavioral assay. Using the startle reflex behavioral assay to evaluate brainstem functioning, both male and female DICE mice demonstrated a larger motor response following various auditory stimuli, as compared to controls. Ongoing studies aim to characterize and quantify A� plaque formation as well as gliosis and microglial expression in brains from DICE mice as compared to APP/PS1/CRE controls using immunohistochemistry, western blot, ELISA, and qPCR. These findings will provide valuable insights into the etiology of Alzheimer’s disease, especially as it pertains to the role of epigenetics.
Approximately 80% of therapeutic monoclonal antibodies are produced by Chinese Hamster Ovary (CHO) cell lines which have been optimized in studies involving nutrient profiling and genomic modification to maximize performance in culture. However, changes in metabolites between culture states and how they are related to product quality and quantity have not been well characterized. CHO cell lines can rapidly proliferate utilizing glucose as a main source of energy. This metabolic state is known as the Warburg effect and leads to high levels of lactate production by the cell. The high accumulation of lactate and cellular debris in culture can hinder cell growth in upstream production and negatively impact downstream protein purification by clogging filters. Previously, our team achieved high protein yield through a design-of-experiment (DoE) approach by studying feed strategies and process parameters, but a fundamental understanding of the inner metabolic state of the cell could lead to more accurate predictions on how the desired cell state can be achieved and maintained. Therefore, it is essential to trace the distribution and flux of downstream metabolites. This project proposes a model study using stable isotopes 1,2 13C2-Glucose and 1,6 13C2-Glucose to trace our in-house VRC01 cell line for metabolic flux analysis (MFA) by utilizing LC-MS. This linkage of intracellular data with product quality information may be used to understand and develop potential intracellular control strategies to improve and maintain product quality attributes of monoclonal antibodies during commercial manufacturing.
Hepatocellular carcinoma (HCC) is one of the most common forms of liver cancer worldwide and accounts for 90% of liver cancer cases. By 2025, it is estimated to have more than one million cases globally. Hepatitis B virus and hepatitis C virus are the main risk factors for HCC development. Early-stage HCC patients may profit from multiple treatment options including surgical resection, liver transplantation, arterial embolization, radioembolization, systemic targeted agent, or liquid biopsy. For patients with advanced HCC, sorafenib is the only approved therapy. The treatment of HCC depends on the tumor stage, patient performance status, and liver function reserved. However, only a small percentage of patients respond well to these treatments, and recurrent diseases remain high. Currently, there is a limited effective option in treating HCC patients. There is an urge to develop more effective therapies for HCC. However, the challenge remains due to the complexity of HCC pathogenesis and the lack of understanding of the complex mechanism. In recent years, the ubiquitin-proteasome system (UPS) and deubiquitinating enzymes (DUBs) have emerged as an important topic for understanding the mechanism of HCC and other cancers. The ubiquitin-proteasome system is one of the most important post-translational modification pathways in eukaryotic cells that regulates cell functions including cell cycle progression, DNA repair, and signal transduction. Ubiquitination can be reversed by deubiquitinating enzymes (DUBs) such as ubiquitin specific peptidase 2 (USP2) that can cleave ubiquitin or ubiquitinated proteins from targeted proteins. As a DUB, USP2a play a critical role in protein degradation and its mRNA and protein level was found to be unregulated in HCC tumor. In addition, USP2a exhibited oncogenic activities by promoting cell proliferation, colony formation and wound healing. Through an unbiased coimmunoprecipitation (Co-IP)-couple proteomic analysis and Western blot of USP2a overexpression pull down results in novel USP2a target proteins involved in cell proliferation, apoptosis and tumorigenesis including HSPA1A, DNAJA1, TCP1, RUVBL1, PCNA. TARDBP and VDAC2. Modulating USP2a expression or its downstream activities can be the molecular basis for developing therapies for HCC.
Peroxisome proliferator-activated receptor alpha (PPARɑ) activates multidrug resistanceassociated protein 3/ATP binding cassette subfamily C member 3 (MRP3/ABCC3) in human hepatocytes
Gina M. Gallucci, Colleen M. Hayes, David N. Assis, James L. Boyer, Nisanne S. GhonemMultidrug resistance protein 3 (MRP3)/ATP-dependent transporter of the ATP-binding cassette family 3 (ABCC3) is a transporter located along the basolateral membrane of hepatocytes, that transports glucuronide metabolites, including bilirubin and bile acid-glucuronides, out of the liver and into systemic circulation. Bile acid glucuronidation is a phase II pathway of detoxification that conjugates glucuronic acid to parent bile acids which results in compounds that are less cytotoxic and more readily excreted. Bile acid-glucuronides are substrates for MRP3, which transports them out of the liver and into systemic circulation for subsequent renal excretion. As such, and due to its role in bile acid detoxification, MRP3 may be a therapeutic target for the treatment of cholestatic liver diseases, which are characterized by bile acid accumulation and toxicity. The peroxisome proliferator-activated receptor alpha (PPARα) ligand fenofibrate upregulates bile acid-glucuronides in the serum of cholestatic patients and upregulates MRP3 messenger RNA in cultured primary human hepatocytes. However, it is unknown how MRP3 is transcriptionally regulated. This study investigates the transcriptional regulation of MRP3/ABCC3 by PPARα. The MRP3 mRNA and protein expression in HepG2 cells were both significantly increased after treatment with fenofibrate (50, 125 µM), compared to vehicle control. In silico analysis of the MRP3 promoter identified multiple PPARα response elements (PPREs). Direct binding of human PPARα to the MRP3/ABCC3 promoter was demonstrated by electrophoretic mobility shift (EMSA) assays. Further investigation will include the activation and binding of PPARα to the MRP3 promoter in vivo by ChIP. Thus far, these data demonstrate that fenofibrate directly binds to functional PPREs within the MRP3 promoter, which contributes to the mechanistic understanding of adjunct fenofibrate for the treatment of cholestatic liver diseases.
As the global population continues to rise, so too has the consumption of material goods. One of the most common commodities on the market in recent decades is plastics, with their global production reaching 460 million tons in 2019. Despite the societal advancements plastics have allowed, the mismanagement of plastic waste has become a pressing global issue, especially the leakage of microplastics.Microplastics (plastic particles <5mm in size) have been shown to induce health issues such as oxidative stress, inflammation, and decreased cell viability in marine organisms. Current research suggests that these microplastics may be transported throughout the environment, however research into their health effects, especially in mammals, is still limited.
This has led our group to explore the biological and cognitive consequences of microplastics exposure in a rodent model. Following a three week exposure to water treated with fluorescentlylabeled pristine polystyrene beads, C57/BL6J mice were assessed using behavioral assays such as open field, followed by tissue analyses such as Western Blot, qPCR, and immunohistochemistry. Data from these assays suggests that short term exposure to microplastics induces both behavioral changes and alterations to immune markers in liver and brain tissues. Additionally, we noted that these changes seem to differ depending on age, indicating a possible age-dependent effect. Furthermore, preliminary studies in a humanized APOE3/4 knock-in mouse model, which may be used to study genetic predisposition to Alzheimer’s disease (AD), suggest that microplastics may be exasperating genetic risk factors of AD in a possibly sex-dependent manner. These findings suggest the need for further research to better understand the mechanisms by which microplastics may induce physiological and cognitive changes.
Pseudoalteromonas piscicida JC3 is a marine bacterial strain with the capacity to produce a large number of membrane vesicles (MVs) as well as antibacterial specialized metabolites. The strain has also been suggested to prevent infection of a deadly pathogen Vibrio parahaemolyticus in shrimps. The mechanism of this probiotic activity is however not completely understood. In this study, we report that JC3 produces a large amount of MVs which may mediate its probiotic effect against V. parahaemolyticus. The production of the MVs involved the membrane filtration of a centrifuged 48 h culture of P. piscicida JC3, and the ultracentrifugation of the resulting filtrate. The MVs size distribution was determined by dynamic light scattering (DLS). They were morphologically characterized by transmission electron microscopy (TEM) and assayed for antimicrobial activity against V. parahaemolyticus PSU5579 and Staphylococcus aureus using the agar spot plate method. The protein and lipid concentrations were determined by Bicinchoninic acid (BCA) protein assay and Stewart assay respectively. The methanolic extract of the MVs was analyzed with liquid chromatography-tandem mass spectrometry (LC-MS/MS) and dereplicated using MS/MS data work-up computer tools. The DLS measured an average diameter of 188.3 nm. The TEM revealed the spherical shape of the vesicles and showed that the MVs had only one membrane, making them outer membrane vesicles. Protein and lipid concentrations of the MVs suspension were 235 ± 6 mg/mL and 741.2 ± 17 µg/mL respectively. The antimicrobial assay done in replicates showed a clear zone of inhibition around the spot the MV suspension was dropped in the models tested. The metabolomic profiling of the membrane vesicles revealed the presence of alterochromides in the cargo of these vesicles. These findings suggest that MVs may contribute to the probiotic activity of P. piscicida JC3 in shrimp aquaculture, and also have potential as sources of antimicrobial compounds.
Lung cancer, one of the leading causes of cancer-related mortalities worldwide is usually diagnosed at an advanced stage, thus necessitating rigorous and effective treatment upon diagnosis. Inhalable nanoparticle formulations facilitate pulmonary delivery of therapeutics in a site-specific and sustained manner. Polymeric nanoparticles (NPs) are being widely investigated owing to their biodegradability, biocompatibility and sustained drug release characteristics at a rate corresponding to the polymer degradation rate. Currently, there is great emphasis on developing multiple polymer or lipid layers incorporated within a single system to improve the distribution and half-life of the formulations, and for combination therapy. However, there are limited studies evaluating the physicochemical properties of these single polymer and multipolymer/lipid formulations post-nebulization. Previously, we demonstrated that polymeric NPs coated with the commercial lung surfactant (LS) Infasurf® can evade phagocytosis by alveolar macrophages in vitro. The present work focuses on evaluating physicochemical properties and in vitro therapeutic efficacy of the LS coated poly lactic-co-glycolic acid (PLGA) NPs after passing through a commercial nebulizer Aeroneb®. Particle size, ζ potential and polydispersity index (PDI) of the NPs were found to be 140.6 ± 3.9 nm, -33.0 ± 1.6 mV and 0.298 ± 0.04 respectively pre-nebulization and 142.1 ± 10.81 nm, -31.4 ± 2.9 mV and 0.261 ± 0.01 respectively postnebulization. FTIR analysis and Stewart’s assay confirmed the retention of the LS layer postnebulization. The nebulization efficiency, nebulization time and fluid output rate of the commercial nebulizer Aeroneb® were determined to be 89.03 ± 3.0 %, 420 ± 57 secs and 261.99 ± 45.1 respectively. The drug release study demonstrated an initial burst release followed by a sustained release in PBS at 37°C. Overall, nebulization did not significantly comprise the physico-chemical properties as well as therapeutic efficacy of the drug loaded NPs.
Nuclear pore complexes (NPC) are the gateway for RNAs and proteins to shuttle between the nucleus and cytoplasm. NPC impairment is often associated with alterations to the nuclear lamina and membrane, as well as to protein mislocalization due to compromised nucleocytoplasmic transport. These phenotypes are consistent with cellular pathologies seen in neurons from patients with the fatal neurodegenerative disease ALS/FTD (Amyotrophic lateral sclerosis/frontotemporal dementia). Thus, we hypothesized that NPC injury can alter activation of transcriptional pathways in ALS/FTD neurons, particularly following stimulatory signals. We found that induced pluripotent stem cell (iPSC)-derived cortical neurons carrying mutations in the C9ORF72 (C9) gene display NPC impairment, altered nuclear membrane morphology, and increased chromatin compaction in an age-dependent manner. However, we found early impairment in the activation of a critical pathway in neuronal health and survival, CREB, in C9 mutant neurons through immunofluorescence and high throughput RNA sequencing assays. Altogether, this data points to the role of not only NPC and nuclear defects, but also of early transcriptional alterations in C9 mutant cells as key and early drivers of pathology.
Aging is associated with diminished immune system function, which renders old people vulnerable to influenza infection and also less responsive to influenza vaccination. This study explored whether the CpG 1018 adjuvant was effective in enhancing influenza vaccine efficacy in aged mice equivalent to human beings in their late 50s to early 60s. Using the influenza pandemic 2009 H1N1 (pdm09) vaccine as a model, we found that the CpG 1018 adjuvant could significantly enhance the pdm09 vaccine-induced serum antibody titer, while the pdm09 vaccine alone failed to elicit significant antibody titer. In contrast, the pdm09 vaccine alone elicited significant antibody titer in young adult mice. Antibody subtype analysis found that the pdm09 vaccine alone elicited Th2-biased antibody responses in young adult mice, while incorporation of the CpG 1018 adjuvant promoted the elicitation of potent Th1-biased antibody responses in aged mice. The pdm09 vaccine alone was further found to induce significant expansion of Th2 cells in young adult mice, while incorporation of the CpG 1018 adjuvant stimulated significant expansion of Th1 cells in aged mice. The CpG 1018 adjuvant also stimulated vaccine-specific cytotoxic T lymphocytes in aged mice. The pdm09 vaccine in the presence of CpG 1018 elicited significant protection against lethal viral challenges, while the pdm09 vaccine alone failed to confer significant protection in young adult or aged mice. Our study provided strong evidence to support the high effectiveness of the CpG 1018 adjuvant to boost influenza vaccination in aged mouse models.
Radiofrequency adjuvant (RFA) was recently developed to boost influenza vaccination without the safety concerns of chemical adjuvants due to their physical nature. Yet, the action mechanisms of RFA remain largely unknown. Omics techniques offer new opportunities to identify molecular mechanisms of RFA. This study utilized comparative tissue proteomics to explore molecular mechanisms of the physical RFA. Comparison of RFA and chemical adjuvant (Alum, AddaVax, MPL, MPL/Alum)-induced tissue proteome changes identified 14 exclusively induced proteins by RFA, among which heat shock protein (HSP) 70 was selected for further analysis due to its known immune-modulating functions. RFA showed much weakened ability to boost ovalbumin and pandemic influenza vaccination in HSP70 knockout than wild-type mice, hinting crucial roles of HSP70 in RFA effects. This study supports comparative tissue proteomics to be an effective tool to study molecular mechanisms of vaccine adjuvants.
Carbonyls are reactive compounds generated from the metabolism of carbohydrates, lipids, and amino acids, and can react with DNA to form glyoxal (GO) / methylglyoxal (MGO)-DNA adducts, causing oxidative DNA damage1. GO and MGO can also disrupt cell membranes, enzymes, and signaling pathways, leading to elevated oxidative stress and various forms of cell death, including ferroptosis2. Ferroptosis is a recently discovered iron-dependent form of cell death characterized by accumulated lipid peroxidation3. Herein, we aimed to depict the overall impact of GO and MGO on erastin-induced ferroptosis in human keratinocytes and investigated through a proteomics-based approach to identify differentially expressed proteins and pathways associated with pathological skin conditions.
Membrane vesicles (MVs) are nanosized single- or double-membraned particles produced by bacteria through blebbing and, for some, explosive cell rupture. MVs may contain biologically active components, such as lipopolysaccharides (LPS), phospholipids, and outer membrane proteins (OMPs), as well as periplasmic components, allowing MVs to be involved in cell-to-cell communication, stress responses, antimicrobial resistance, horizontal gene transfer, and immune evasion. MVs can also package virulence factors and may function as delivery vesicles, contributing to host colonization and infection-associated pathology. MVs have been proposed as a vaccine platform technology to limit the severity of infectious disease outbreaks. In this study, MVs were isolated and purified from the marine bacteria Pseudoalteromonas piscicida JC3, Vibrio corallilyticus RE22, Vibrio parahaemolyticus PSU5579, and Phaeobacter inhibens S4. The MVs were subjected to an array of in vitro assays to assess their possible roles in microbial interactions. Spot plate antibacterial assays showed that P. piscicida JC3 possesses antibacterial potential against P. piscicida PSU5579 and Staphylococcus aureus. MVs from V. parahaemolyticus PSU5579 were found to reduce biofilm production by P. inhibens S4. Additionally, MVs from P. piscicida JC3 and P. inhibens S4 demonstrated potential for iron acquisition via siderophore production. These results demonstrate the production of MVs in phylogenetically diverse marine bacteria and indicate possible ecological roles for mediating microbial interactions and nutrient acquisition.
Background: Fibromyalgia is a condition associated with pain, fatigue, sleep disturbance, and emotional and cognitive distress. Over 4 million individuals are affected by fibromyalgia in the US. Fibromyalgia is more prevalent in women compared to men(1). Naltrexone HCl is an opioid antagonist approved for treating alcohol and opioid use disorders. Low-dose naltrexone (LDN) is defined as Naltrexone HCl doses <10mg. LDN is currently used as an off-label medication in the management of fibromyalgia.
Method: This study was performed following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines in March -April 2023. Electronic databases (PubMed, Embase, Scopus, Cochrane Library) and gray literature (ProQuest, Clinicaltrial.gov) were searched using “Fibromyalgia” and “Low-dose naltrexone” search terms.
Results: 565 articles were identified from the database search. 8 articles (6 full papers and 2 abstracts) were identified for inclusion in the systematic review and meta-analyses (Five RCTs, and Three Observational Studies). In the analysis of pain score, there was a significant reduction in the post-LDN group relative to the baseline pain score by 0.72 [95%CI: -1.21, -0.22; I2= 64%]. Pain score was reduced by 0.31 in the LDN group compared to the placebo group [95%CI: -0.61, -0.01; I2=0%]. In the analysis of FIQ scores, the FIQ score was reduced by -1.08 [95%CI: -1.62, -0.55; I2=63%] post-LDN relative to the baseline. There was a non-significant reduction in FIQ score by 0.46 [-1.48, 0.56; I2=86%] in the LDN group compared to the placebo. In the subgroup analysis by full-text publications, the Mean Pain score and FIQ score were reduced by 0.59 [95%CI: -1.04, -0.14; I2=0%] and 0.82 [95% CI:-1.21, -0.42; I2=0%] respectively comparing the post-LDN to the pre-LDN.
Conclusion: Low-dose naltrexone is significantly associated with a reduction in pain and FIQ scores.
Uncovering the anti-inflammatory mechanisms of phenolic-enriched maple syrup extract in lipopolysaccharide-induced peritonitis in mice: Insights from data-independent acquisition proteomics analysis
Tess Puopolo, Huifang Li, Hang Ma, Joseph Schrader, Chang Liu, Navindra P. SeeramOur group has previously reported on the phytochemical composition and biological activities of a phenolic-enriched maple syrup extract (MSX), which showed promising anti-inflammatory effects in several disease models including diabetes and Alzheimer’s disease. However, the efficacious doses of MSX and its molecular targets involved in the anti-inflammatory effects are not fully elucidated. Herein, the efficacy of MSX in a peritonitis mouse model was evaluated in a dose-finding study and the underlying mechanisms were explored using data-independent acquisition (DIA) proteomics assay. MSX (at 15, 30 and 60 mg/kg) alleviated lipopolysaccharide-induced peritonitis by reducing the levels of pro-inflammatory cytokines including interleukin-1 beta (IL-1β), IL-6, and tumor necrosis factor alpha (TNF-α) in the serum and major organs of the mice. Furthermore, DIA proteomics analyses identified a panel of proteins that were significantly altered (both up- and down-regulated) in the peritonitis group, which were counteracted by the MSX treatments. MSX treatment also modulated several inflammatory upstream regulators including interferon gamma and tumor necrosis factor. Ingenuity pathway analysis suggested that MSX may modulate several signaling pathways in the processes of initiation of cytokine storm, activation of liver regeneration, and suppression of hepatocyte apoptosis. Together, these proteomic and in-vivo findings indicate that MSX could regulate inflammation signaling pathways and modulate inflammatory markers and proteins, providing critical insight into its therapeutic potential.
Francisella tularensis is a highly infectious, intracellular human pathogen that can cause fatal disease. The F. tularensis type VI secretion system is an essential virulence factor required for survival in host cells, including survival in what are considered a key niche, macrophage. We have determined that ribosome composition influences production of the F. tularensis type VI secretion system and virulence. Ribosomes containing one of the three homologs for the small ribosomal subunit protein bS21, bS21-2, positively control key virulence genes and intramacrophage replication. However, the mechanisms that control and coordinate production of bS21-2 and the other bS21 homologs, bS21-1 and bS21-3, are unknown. We have found that bS21-2 negatively regulates its own production, as the presence of bS21-2 leads to significant reductions in abundance of its transcript, rpsU2. Further, we have found that the 5′ untranslated region (UTR) of rpsU2 is sufficient for this bS21-2-mediated repression. Production of bS21-2 appears to be tightly controlled by bS21 levels in the cell, as both bS21-1 and bS21-3 also negatively regulate bS21-2 transcript abundance. In contrast, bS21-1 and bS21-3 do not affect their own production. Thus, the bS21-mediated regulation of bS21-2 appears to be unique among the three homologs. This suggests that F. tularensis integrates multiple signals into a regulatory network to control the appropriate production of each bS21 homolog. This regulatory network in turn may control ribosomal heterogeneity and virulence gene expression.
Background: Increased national attention is directed to opioid prescribing among generalist physicians.
Methods: We analyzed the Medicare Part D Prescribers by Provider and Drug dataset to determine the number of opioid claims per Medicare enrollee for US 2020 and the percentage of opioid claims by provider type.
Results: Part D prescription opioid claims ranged from 0.566 to 2.129 per enrollee across US states during 2020. Alabama led among 50 US states the with highest annual rates of Part D prescription opioid claims for the year 2020. Whereas Hawaii shows the lowest rates among 50 states annual rates of Part D prescription opioid claims. Higher rates of prescription opioid claims were observed for the US South among the 4 US census regions. General medical practitioners prescribed opioids approximately as many as 3 times higher than those specialty practitioners in Vermont, Iowa, Maine, Mississippi, and Tennessee. Vermont had the highest percentage of opioids issued by generalist providers at 76%. Florida had the lowest percentage of opioids issued by generalist providers at 38%.
Conclusion: Using a national sample of Medicare beneficiaries, we observed that during the year 2020, the use of prescription opioid claims was highest in Alabama and lowest in Hawaii, while the percentage of opioids prescribed by generalist providers varied widely across US states.
Flagellin (FH) platform has been used as to perform hybrid by replacing the surface exposed D3 domain of flagellin. FH virus like particles were prepared by the insertion of flagellin gene into the c/el loop of HBC. This gene has been expressed in the E. coli expression, purification and self-assembly into VLPs. FH-VLP based M2e peptide vaccine elicited more potent anti-M2e antibody responses and conferred significant protection against multiple influenza A viral strains. FH VLP based OVA peptide vaccine elicited more potent CTL responses and protection against lymphoma and melanoma challenges. The preclinical data shows FH VLPs to be highly immunogenic and versatile platform. Our present work is to increase the total amount of FHM2e and producing endo free vaccine without losing mass amount of protein.
Bret Abreu PharmD. Candidate, Celina Chau PharmD. Candidate, Jack Dolan PharmD. Candidate, Nick Mcpadden PharmD. Candidate, Rebecca O’Toole PharmD. Candidate
Purpose:
Current data identifies uncontrolled diabetes and elevated hemoglobin A1C (HbA1C) as risk factors for chronic kidney disease (CKD)1,2,3 Supporting patients’ in meeting HbA1C goals is essential to improving health outcomes and preventing CKD2,3. Patients with diabetes and CKD are at significantly increased risk for cardiovascular disease 1,3,4, Previous studies show that CKD does not impact HbA1C 5. Our goal is to determine if a relationship between decreased renal function and ability to meet HbA1C goals exists.
Methods:
This cross-sectional study utilized data collected from the 2017-2018 National Examination Survey (NHANES). Decreased renal function was defined as creatinine clearance (CrCl) less than 60mL/min via the Cockcroft-Gault equation. The outcome was defined as a measured HbA1C above the goal. Male patients aged 18-79 years old with a diagnosis of diabetes were included and patients with missing/unknown values were excluded; yielding a total of 226 patients. A Chi square test and multivariable logistic regression with a one sided T-test (ɑ = 0.05) was conducted using SPSS to evaluate whether decreased kidney function impacts success in meeting HbA1C goals.
Results:
84% of patients (N=190) had a CrCl above 60mL/min and 15% (N=36) had a CrCl below 60mL/min. 87% (N=128) of patients with a CrCl above 60mL/min and 52% (N=19) of patients with CrCl below 60mL/min, did not meet their HbA1C goal. Patients with a CrCl below 60 ml/min are 58% less likely to reach their HbA1C goal (95% CI 0.151-1.167; p = 0.096).
Conclusion: There was no significant difference in the odds of achieving target A1C levels for males between the age of 18-79 years old in those who have poor renal function compared to those who do not have poor renal function.
Purpose:
The nicotine in cigarettes, e-cigarettes, and tobacco may increase blood pressure (BP) and contribute to hypertension. Amlodipine, a calcium channel blocker (CCB), and lisinopril, an angiotensin-converting enzyme inhibitor (ACE-I) are first-line treatment options for hypertension. The objective of this study was to compare the clinical efficacy of amlodipine and lisinopril among patients with hypertension that smoked or used smokeless tobacco through BP values.
Methods:
This national cross-sectional retrospective study included patients taking amlodipine or lisinopril from the National Health and Nutrition Examination Survey (NHANES) of the 20172018 cycle. Inclusion criteria included being diagnosed with hypertension, defined as BP values over 130/80 mmHg and smoking cigarettes, e-cigarettes, or using smokeless tobacco. Exposure was defined as the use of either amlodipine or lisinopril. Outcome was defined as the efficacy of these medications, indicated by BP. A logistic regression analysis adjusted for multiple covariates was used to compare BP values and obtain an odds ratio among both amlodipine and lisinopril users.
Results:
253 individuals diagnosed with hypertension were included, of which 205 were taking amlodipine and 48 were taking lisinopril. Among the amlodipine and lisinopril users, 68 (33.2%) and 21 (43.7%) had BP values less than 130/80, respectively. The odds ratio of 0.673 suggested that lisinopril users were about 33% less likely to have hypertension. However, the p-value (significance level = 0.05) of 0.167 suggested that there was no significant difference between the two medications.
Conclusion:
Although the study demonstrates that lisinopril may have better anti-hypertensive effects, a future study with a larger sample size is warranted to demonstrate an effect. Limitations to the study included not incorporating data on other medications, not identifying all potential risk factors and covariates, and not having data on specific BP values before and after using the medications.
Purpose:
Levothyroxine (Synthroid) is the most common medication prescribed for the treatment of hypothyroidism and the second most commonly prescribed medication in the US1, 2. However, overreplacement of thyroid-stimulating hormone potentiates an association with decreased bone mineral density (BMD), putting patients at a higher risk of developing osteoporosis3. Some studies have been performed, but results have been inconclusive4, 5 . The purpose of this study is to determine if levothyroxine use puts patients at an increased risk of developing osteoporosis, based on their femoral neck BMD.
Methods:
This cross-sectional study used data from the 2017-2018 National Health and Nutrition Examination Survey (NHANES). A logistic regression was run that adjusted for confounders. A Pearson’s Chi Square test or Fisher’s Exact test (where counts ≤5) were used to obtain P values for categorical variables. A one sided T test was used to obtain P values for continuous variables. The significance level for all tests was 0.05.
Results:
Among the 2,014 participants who received a DXA scan, 5.1% reported taking levothyroxine (n=103). In the levothyroxine group, 75.7% (n=78) were female and 24.3% (n=25) were male ( p <0.001). The average femoral neck BMD in patients taking levothyroxine was 0.75g/cm2 and 0.77g/cm2 in patients not taking levothyroxine (p <0.001). More patients on levothyroxine had a lower BMD (68% vs. 60.9%), however, the results were not significant (p = 0.152). There was no significant increase in risk of osteoporosis development in those who use levothyroxine (OR: 1.096, 95% CI: 0.696-1.723).
Conclusions:
These results suggest that levothyroxine does not affect BMD while also accounting for age, gender, and past medical history. These findings are consistent with the limited studies that have been published, however, more studies are needed for confirmation.
Purpose:
Studies evaluating the rates of psychological triggers in patients with asthma and concurrent anxiety are limited. Existing data has demonstrated that anxiety and symptoms of anxiety in asthmatic patients were associated with higher rates of uncontrolled asthma and poorer quality of life scores. This study aimed to determine if anxiety increases asthma exacerbation rates in asthmatic patients.
Methods:
A cross-sectional survey design was used with data collected from the 2017-2018 NHANES questionnaire. Inclusion criteria included participants over the age of 18 that had a diagnosis of asthma with or without medication therapy for anxiety. Exclusion criteria included participants without an asthma diagnosis and those with asthma who did not have feelings of “worried, nervous, or anxious”. A binary logistic regression was used and adjusted for multiple covariates to generate an odds ratio (OR) and compare against the exposure group with 95% confidence intervals. A Pearson's Chi-square test and a paired t-test was used to generate P values for categorical variables and continuous variables, respectively.
Results:
Among 102 participants that took medications for anxiety, 54.9% (56/102) experienced an asthma attack in the past year compared to 39.2% (169/431) for participants who did not take medications for anxiety. The risk of experiencing an asthma attack in the past year in patients medicated for anxiety was 1.89 times (95% CI 1.22 - 2.92; p-value = 0.004) higher than those non-medicated for anxiety.
Conclusion:
The results of the study were statistically significant showing that patients taking medication for anxiety were more likely to have had an asthma attack within the last year. Limitations included using a cross sectional study, unknown medication adherence, recall bias, and unknown definitive anxiety diagnosis. Future research correlating anxiety severity and medication therapy to asthma exacerbations are needed to further strengthen the conclusion
Purpose:
Approximately 12.5% of US adults currently use tobacco. Smoking increases LDL-C, an important marker for CVD, increasing risk for adverse cardiac events. Literature addresses effects of cigarettes, e-cigarettes, and smokeless tobacco separately on LDL-C. This study groups cigarettes, e-cigarettes, or smokeless tobacco users' effects on LDL-C levels.
Methods:
2017-2018 NHANES data was used. A chi-square, independent t-test, and multivariable linear regression were conducted using SPSS. Those who said yes to using cigarettes, e-cigarettes, or smokeless tobacco and whose LDL-C levels were recorded using the Friedewald equation (mg/dL) were included. Exclusions included those who said no to using cigarettes, e-cigarettes, or smokeless tobacco and whose LDL-C wasn't recorded. Covariates included age, gender, BMI, SBP (mmHg), and fasting glucose (mg/dL). The outcome of interest was clinically abnormal LDL-C (>130 mg/dL).
Results:
There’s a significant difference between tobacco and non-tobacco users regarding gender and BMI. There’s no significant difference between LDL-C >130mg/dL and tobacco versus nontobacco users. After controlling for confounders, there’s no significant difference in LDL-C >130mg/dL between tobacco users versus non-tobacco users. It’s significant that each SBP category is more likely to have LDL-C >130mg/dL versus the reference. It’s significant that fasting glucose > 126mg/dL is less likely to have LDL-C >130mg/dL versus the reference.
Conclusions:
There’s no significant difference between LDL-C > 130mg/dL and tobacco users versus nontobacco users. While our data consisted of a large sample, it may not represent the population. We didn’t assess the role diet played on LDL-C, nor if patients took cholesterol medications or exercised. With limited research about impacts of e-cigarettes on LDL-C, this is a future research direction.
Purpose:
Almost 50% of American adults experience hypertension (HTN), 80% being uncontrolled.1,2 Hypertension, defined as SBP >130mmHg or DBP >80mmHg, is linked to an increased risk of adverse cardiovascular events (CVE).2,6 A meta-analysis of 61 prospective studies found increases of 20mmHg SBP or 10mmHg DBP were associated with doubled risk of death from CVE.3,4 The objective of this study was to assess prevalence of hypertension in those who experienced a CVE.
Methods:
A cross sectional study was performed using National Health and Nutrition Examination Survey (NHANES) data of 5,542 US adults enrolled between 2017-2018. A CVE was defined as congestive heart failure, coronary heart disease, angina, heart attack, or stroke. Participants with missing/unknown values were excluded. Multivariable regression was performed to assess covariate influences (age, BMI, diabetes, dyslipidemia, smoking)5 .
Antihypertensive medication impact was excluded due to limited information, requiring further analysis.
Results:
Of 5,542 individuals, 644 patients experienced a CVE with 45% of those having hypertension. Between those experiencing a CVE and not, there were statistically significant differences in baseline characteristics (age, gender, race, BMI, weight, dyslipidemia, diabetes, smoking status).
Initial analysis shows increased prevalence of HTN in those having experienced CVE (OR 1.607, p <0.001). Covariate adjustment demonstrated statistically significant impact of confounders and no significance between prevalence of HTN and CVE (adjOR 1.061, p = 0.550).
Conclusion:
No statistically significant relationship between CVE and prevalence of hypertension was demonstrated following covariate adjustment. Limitations included lack of potential confounder information, confounder prevalence, and data restrictions. Further studies, removing covariates, are needed to determine the relationship between HTN and CVE.
Marina Ayad Pharm.D. Candidate, Evona Bolaske Pharm.D. Candidate, Gianni Lazarides Pharm.D. Candidate, Ji-Woo Lee Pharm.D. Candidate, Sydney Story Pharm.D. CandidatePurpose:
Creatinine clearance is a measure of kidney function and approximates filtering rate of creatinine in the kidneys. Generally, a creatinine clearance below 90 mL/min indicates decreased kidney function. Medications, such as loop and thiazide diuretics, can increase urine production in the body. This study aimed to assess the incidence of urinary leakage in individuals with abnormal creatinine clearance, defined as less than 90 mL/min, who are taking a loop or thiazide diuretic.
Methods:
Data from the National Health and Nutrition Examination Survey (NHANES) was evaluated for individuals with kidney conditions taking diuretic prescription medications between 2017 and 2018. Inclusion criteria included individuals with abnormal CrCl defined as < 90 mL/min taking either a loop or thiazide diuretic. Loop diuretics included furosemide, bumetanide, and torsemide. Thiazide diuretics included hydrochlorothiazide, chlorthalidone, and indapamide. The outcome of interest was incidence of urinary incontinence, defined by self-reported urinary leakage. Statistical tests performed to evaluate the effect of different diuretic classes on urinary incontinence included Pearson Chi-Square tests, single sample t-tests, and logistic regression.
Results:
There were 91 individuals in the study who met eligibility criteria. The logistic regression showed loop diuretic users were approximately 25% less likely (OR = 0.755) to have incidence of urinary leakage compared with thiazide users but yielded p = 0.594 meaning this result was statistically insignificant. Comparing the frequency of urinary leakage within the population yielded p = 0.179 which is also statistically insignificant.
Conclusions:
The study findings concluded the incidence of urinary incontinence with loop and thiazide diuretics were not statistically significant. Limitations for our study included unknown dose, adherence and indications for the diuretics, concomitant medication usage, and a small sample size (n=91). Additional research is required to compare urinary incontinence in individuals with abnormal CrCl taking a loop or thiazide diuretic.
Purpose:
Dyslipidemia and calculated low-density lipoprotein (LDL-C) control is essential for preventing secondary cardiovascular events.1,2,3 There is a demonstrated relationship between absolute changes in LDL-C levels and an increased risk of secondary events2,3. Access to health insurance is a suspected factor for improved management of LDL-C levels4,5. The purpose of this study is to evaluate the relationship between those with and without health insurance reaching target LDL-C levels for secondary prevention.
Methods:
This cross-sectional study utilizes 2017-2018 NHANES data to examine LDL-C control among secondary prevention patients. Patients were included based on answering “yes” for a previous stroke or MI. The study outcome was defined as an LDL-C>100 mg/dL as calculated by the Friedewald formula6. The study exposure was a lack of current health insurance coverage as reported by the participant. Unadjusted and adjusted odds were calculated using a binary logistic regression model. The cutoff for statistical significance is a p <0.05. All statistical analyses were conducted with IBM SPSS.
Results:
The characteristics between the populations are evenly distributed in all characteristics except age, with a median age of 60 and 68. There is a 4.336 (95% CI; 1.628, 11.552) increase in odds of having an LDL-C>100 mg/dL for uninsured patients vs insured patients after a primary MI or stroke (p=0.003).
Conclusions:
The results of this study indicate a statistically significant difference in uncontrolled LDL-C levels for those uninsured compared to those that have health insurance. Some limitations of our study are insurance status and cardiovascular event history both being self-reported, and our uninsured population is significantly smaller compared to insured. This study suggests the importance of insurance in achieving treatment goals as a modifiable risk factor for clinicians and patients to consider.
Chris Ballou PharmD. Candidate, Evan Goncalves PharmD. Candidate, Kathryn Higgins PharmD. Candidate, Will Sullivan PharmD. Candidate, and Justin Trask PharmD. CandidatePurpose: Given the persistent rise of recreational drug use among the American population, the association with the severity of asthma remains uncertain. Some of these recreational drugs can be used via inhalation, such as marijuana, cocaine, and methamphetamine. The objective of this research is to assess the association between inhaled recreational drug use and the prevalence of asthma exacerbations.
Methods:
Using data from the 2017-2018 National Health and Nutrition Examination Survey, a multivariable logistic regression was performed to compare the exposure and outcome. The exposure was use of inhaled recreational drugs, including marijuana, cocaine, and methamphetamine. The outcome was the presence of asthma attacks in the past year. The inclusion criteria consist of patients aged 18-40 years old with current asthma who answered the screening questions regarding marijuana and cocaine/methamphetamine use. The exclusion criteria included patients below 18 and above 40 years-old who do not have current asthma and did not answer all relevant survey questions. The covariates included age, gender, race/ethnicity, eosinophil count, smoking status, education level in adults over 20 years old, and chronic bronchitis status.
Results:
A total of 146 participants met the inclusion criteria, with 83 reporting recreational drug use and 63 reporting no recreational drug use. There were no significant differences between the evaluated clinical characteristics among the study populations. The incidence of asthma attacks in participants who reported recreational drug use was 57.8%, versus 30.2% in those not using recreational drugs. The logistic regression results show a significant association between inhaled recreational drug use and asthma attacks in the past year (P = 0.050) when adjusting for multiple covariates.
Conclusion:
Inhaled recreational drug use was shown to increase asthma attacks in patients aged 18-40 years old, however, more studies are needed to prove a causal relationship between recreational drug use and asthma severity.
Roberto PharmD. Candidate, Blinere Bytyqi PharmD. Candidate, Jia Ying Lin PharmD. Candidate, Avery Salcedo PharmD. Candidate, and Elena Tormo PharmD. CandidatePurpose:
Patients with chronic kidney disease (CKD) may develop anemia due to decreased renal production of erythropoietin as a result of impaired kidney function. The purpose of this study was to analyze the relationship between serum creatinine level and the prevalence of anemia among patients living with CKD.
Methods: Cross sectional data was taken from the National Health and Nutrition Examination Survey (NHANES) to select participants, according to calculated estimated Glomerular Filtration Rate (eGFR), with an eGFR less than or equal to 60 milliliters per minute. Participants were stratified into three SCr subgroups, and coded according to baseline hemoglobin levels. Analysis of anemia prevalence was assessed in each subgroup using an adjusted logistic regression.
Results: Four hundred and forty one participants met inclusion and exclusion criteria. Exposure groups differed for participants' median age, gender, and prevalence of diabetes. Exposure groups were similar for participant’s race and prevalence of thyroid disorder. The study found prevalence of anemia increased among participants with SCr greater than 1.3 mg/dL, when compared to participants with SCr less than or equal to 1.3 mg/dL.
Conclusions: The study findings support an increased prevalence of anemia among CKD patients with higher serum creatinine values. Limitations of the study include limited generalizability because patients without self reported eGFR values were excluded from the population. Reporting bias may also reduce the study’s validity. The study used cross sectional data, limiting the ability to identify a temporal relationship. These results may be used to stimulate further research to understand a causal relationship between anemia prevalence and serum creatinine values.
PharmD. Candidate, Jessica Samuel PharmD. Candidate, Matthew Tocco PharmD. CandidatePurpose:
Osteoporosis is a condition that causes diminishment of bone density, increasing fracture risk.1 Data has shown that patients with Type I and II diabetes may have an increased risk for comorbid conditions like osteoporosis.2 Bone remodeling and turnover is compromised in diabetic patients, with longer duration, poor glycemic control, and thiazolidinone use for glycemic index control as risk factors for fracture.3,4,5 The purpose of this study was to assess the risk of osteoporosis secondary to the use of three antidiabetic medications: insulin, metformin, or sulfonylureas.
Methods:
The study was a retrospective cross-sectional survey study of 4,910 participants using NHANES data. Exposure was defined as patients taking one of three defined drugs, concomitant osteoporosis identified through self-report or total bone mineral density (BMD) 2.5 standard deviations below average. Inclusion criteria: participants over age 25 reporting a HbA1C, osteoporosis, gender, total BMD, alcohol use or smoking. Outcome assessment defined as incidence of osteoporosis. Covariates for the data included age, gender, alcohol use, smoking, and HbA1C level (%). Multivariate logistic regression was used in determination of the results.
Results:
Utilizing a Chi-square test, results showed that all three of the prespecified medications were not statistically significant for the increase in osteoporosis risk as all carried a p-value greater than 0.05. Additionally, the multivariate logistic regression yielded results of age, smokers, alcohol users, and men as statistically significant. An odds ratio of 1.338 was identified for alcohol users, indicative of osteoporosis risk.
Conclusions:
After survey data was assessed, the majority of patients with osteoporosis had no relationship to diabetes. These results do not align with current data regarding risk factors for osteoporosis. Limitations include, temporality, limited diabetes diagnosis data, recall bias, incomplete medication lists, age, quantified alcohol consumption and smoking quantity. Future studies should include duration of treatment for the medications of interest.
Thomas Bokinz Pharm.D. Candidate, Abbigayle Fielder Pharm.D. Candidate, Liam Furlong Pharm.D. Candidate, Abigail Horan Pharm.D. Candidate, and Taya Kerwin Pharm.D. CandidatePurpose:
Arthritis remains one of the leading causes of disability among elderly females in the United States1. Serum creatinine (Scr) can be obtained to assess kidney function. Normal SCr range for men is 0.7-1.3 mg/dL and women is 0.6-1.1 mg/dL2 . Previous literature demonstrated that arthritis can be associated with an increased Scr3,4,5 . The study objective was to assess the impact of arthritis on kidney function by comparing Scr between patients with and without arthritis.
Methods:
We performed a Wilcoxon rank-sum and chi-square test to analyze the covariates and Scr along with a multiple linear regression to assess correlation using the NHANES 2017-2018 data. The primary outcome was Scr, and risk factors included age, gender, race, kidney failure status and recent increase in exercise. Exclusion criteria eliminated patients <20 and ≥80 years-old, not answering for arthritis status, and without data for primary outcome.
Results:
The population consisted of 4,482 patients with a median age of 62 (arthritis) and 44 (without arthritis). At baseline, 6.7% patients with arthritis and 1.9% patients without arthritis had been told they had weak or failing kidneys. The raw comparison showed significant increase in Scr level for patients with arthritis (p=<0.001). However, the multivariable linear regression showed this was not significant when accounting for covariates (p=0.098).
Conclusion:
The study findings suggest that the difference in median Scr between patients with and without arthritis was not significant. Since there are other factors that have effects on Scr and there was a disproportionate patient population between those with arthritis and those without, this data is not applicable to practice. Due to the retrospective analysis, there is no assessment on causality between the exposure, covariates and outcome over time which further limits the relativity of the data.
Harry Braidt PharmD. Candidate, Kyle Gott PharmD. Candidate, Sarah Hughes PharmD. Candidate, Caroline Joncas PharmD. Candidate, Celia Parisi PharmD. CandidatePurpose:
It is known that hypertension and oral hormonal contraceptive use in women are both risk factors for cardiovascular events including stroke and myocardial infarction. The study objective is to assess if there is an association between taking birth control and the frequency of cardiovascular events in the population of females with hypertension, and if it should affect the prescribing of hormonal contraceptives in this population.
Data included in this study was collected from 2017-2018 Continuous NHANES Questionnaire Data. A data analysis was run using SPSS to investigate the incidence of cardiovascular events. The study population from the survey data included all females who were ever told that they had high blood pressure. The incidence of cardiovascular events was compared between subjects who answered yes to ever taking birth control and those who answered no. A chi-square analysis was conducted using a P-value significance level of 0.05 to determine if there is statistical significance in the cumulative incidence of the primary outcome.
Results:
The relative risk when comparing cumulative incidence of cardiovascular events between subjects exposed versus not exposed to hormonal contraception is increased by 3.8% with a Pvalue of 0.839, indicating that the results were not statistically significant. Therefore, we accept the null-hypothesis that there is no association between cardiovascular events and hormonal contraceptives in this population.
The retrospective cohort study findings demonstrate a correlation between use of hormonal contraception and cardiovascular events, although the results were not statistically significant. Therefore, these results should not make an impact on clinical-decision making of prescribing hormonal contraception for females with hypertension. Limitations include a lack of information on the sequence of events including when subjects were told they had hypertension, when they took birth control and when they experienced a cardiovascular event.
Bailey Burke PharmD. Candidate, Cassie Capezza PharmD. Candidate, Robert Davis PharmD. Candidate, Cassidy Lawrence PharmD. Candidate, Olivia Violette PharmD. CandidatePurpose:
Type II diabetes mellitus increases the risk of cardiovascular disease as well as morbidity and mortality. This study assesses the effect of a previous myocardial infarction (MI) on current glycemic control in patients with type II diabetes based on current glycohemoglobin values.
Methods:
The NHANES data used in this study was obtained from a cross sectional survey of participants between 2017 and 2018. Patients 50 years or older with type II diabetes mellitus who had a previous MI (n=91) and who have no history of MI (n=537) were included in this study. The exposure of interest was a prior MI, and the outcome of interest was lab-reported glycohemoglobin dichotomized as controlled (≤ 7%) or uncontrolled (>7%). Using logistic regression, an odds ratio was generated to evaluate our exposure of interest. Non-parametric independent t-tests and chi-square tests were used for continuous and categorical variables respectively with a significance level of 0.05.
Results:
A total of 628 patients meeting inclusion criteria were compared. In the group with a prior MI, 54 patients had controlled HbA1c while 37 had uncontrolled HbA1c. The group with no prior MI consisted of 275 patients with controlled and 262 patients with uncontrolled HbA1c. Based on raw data, the difference in glycohemoglobin control for patients with or without a previous MI was not statistically significant with a p-value of 0.151. Logistic regression analysis also showed no statistical significance between prior MI and HbA1c control while accounting for multiple confounders (age, gender, total cholesterol, smoking, and systolic blood pressure) (OR 1.28 [0.782, 1.984] p = 0.356).
Conclusions:
This study found that in patients with type II diabetes older than 50 years, there was no significant difference in glycemic control based on myocardial infarction exposure.
Ashley Chiarello, Pharm.D. Candidate, Victoria Gugliotti, Pharm.D. Candidate, Kimberly Khan, Pharm.D. Candidate, Krystle Sclafani, Pharm.D. Candidate, Christine Wu, Pharm.D. Candidate
Dyslipidemia is an imbalance of cholesterol, low-density lipoprotein (LDL), triglycerides, and high-density lipoprotein in the blood.1 Statins, the first-line therapy for treating dyslipidemia, lower LDL by slowing down cholesterol production.2 Although cigarette smoking is an established risk factor for dyslipidemia, there is no conclusive data regarding the association of statins with smoking e-cigarettes and marijuana.3 This study aimed to assess the efficacy of statin therapy on LDL in patients with the composite outcome of inhaled substance use consisting of smoking tobacco, e-cigarettes, and marijuana.
The cross-sectional National Health and Nutrition Examination Survey data from 2017-2018 was used to assess the primary outcome of LDL dichotomized as “high” (> 100 mg/dL) or “not high” (≤ 100 mg/dL).4 Patients included in the study had a prescription for rosuvastatin, atorvastatin, lovastatin, pravastatin, or simvastatin for their dyslipidemia. A logistic regression analysis adjusted for multiple covariates (age, gender, BMI, race, current alcohol use, and statin intensity) to generate an odds ratio comparing the exposures of interest, with 95% confidence intervals.
There were 130 participants in the sample size comparing dyslipidemic patients with a history of inhaled substance use (n = 23) and without (n = 107). Patients with a history of inhaled substance use had 0.417 times the risk of high LDL compared to the unexposed group (p = 0.139).
The results suggest that there is no statistically significant difference in statin efficacy on LDL values between the two groups. Causation between the exposure and outcome cannot be determined due to the lack of power in the study and use of cross-sectional design.5 More robust studies must be conducted to conclude whether a history of inhaled substance use affects LDL levels before treatment guidelines are amended.
Purpose:
Diabetic peripheral neuropathy (DPN) is a common complication in patients with diabetes (70%)1, serving as the leading cause of amputation amongst these individuals.2 First line pharmacotherapy includes anticonvulsants and antidepressants.3 Anticonvulsants inhibit calcium channels, decreasing excitatory neurotransmission.4 Antidepressants inhibit reuptake of serotonin and norepinephrine, enhancing analgesic effects.5 Studies assessing neuropathy medication use in patients with diabetes with a low A1C compared to a high A1C are limited in literature.6 The objective of the study was to determine whether an association exists between A1C levels and use of neuropathy medications in patients with diabetes.
Methods:
A cross-sectional study was designed based on data collected from the 2017-18 National Health and Nutrition Examination Survey. We included participants with a confirmed diabetes diagnosis and an obtained A1C value. The exposure was an A1C > 8% (compared to A1C < 8%), and the primary outcome was whether participants reported taking neuropathy medications. Pearson's Chisquared, independent T-test, and logistic regression were used for analysis.
Results:
The sample size for the analysis included 734 participants, 335 of which had a high A1C. There were no significant differences between the exposure groups in terms of neuropathy prescription treatment. Approximately 8% of patients in each group were on neuropathy medications. Duloxetine (4.9%) gabapentin (2.2%), and amitriptyline (1.5%) were the medications reported. Patients with a low A1C were found to have a positive, yet nonsignificant association with being on neuropathy medications (OR = 1.075, P value = 0.859).
Conclusions:
Findings do not support the causal relationship between A1C levels and increased likeliness of neuropathy treatment. Future research utilizing a cohort study design is indicated to determine whether a stronger causal relationship exists. Study limitations include the inability to determine causation, small sample size, unknown confounders, and recall bias.
Purpose:
Cocaine, heroin, and methamphetamine are some of the most commonly used illicit substances in the US.1 These substances result in a dopamine increase that may alter blood pressure.2,3 Currently, there is limited evidence evaluating the effects of illicit substances on blood pressure.2 The purpose of this cross-sectional study was to assess the effects of cocaine, heroin, methamphetamine on blood pressure regulation among patients with established hypertension.
Methods:
We utilized data from the NHANES 2017-2018 Questionnaire. Inclusion criteria required answering “yes” to “ever been told you have high blood pressure?” Participants were excluded if they answered “no”, if it was not answered, or if a systolic blood pressure (SBP) reading was unavailable. A logistic regression model calculated odds ratios, adjusting for sex, age, BMI, and race. An independent t-test determined the significance between illicit drug use and continuous variables (age, systolic blood pressure, diastolic blood pressure) and a Chi-square determined the impact of illicit drugs on hypertension. Lack of proper blood pressure control was defined as SBP ≥ 130 mmHg.
Results:
Those that reported illicit drug use had a nearly identical odds of SBP ≥ 130 mmHg compared to those that did not use illicit drugs, but this finding was not statistically significant (adjusted OR: 1.067; 95% CI (0.779-1.461); p = 0.686). Increasing age was associated with a higher odds of SBP ≥ 130 mmHg.
Conclusion: There was overall no significant difference in blood pressure between people who have used illicit drugs and those who have not among patients with established hypertension. Potential confounders include smoking status and medications used to treat hypertension. Further research should be conducted to prove an association between illicit drug use and hypertension control.
Danthi Chu PharmD. Candidate, Sarah Cunningham PharmD. Candidate, Jenna D’Alessio PharmD. Candidate, Colin Lavigne PharmD. Candidate, Toni Sullo PharmD. CandidateEmma Daly Pharm.D. Candidate, Alex DiLucia Pharm.D. Candidate, Kaitlyn Goncalves Pharm.D. Candidate, Taylor Mezini
Pharm.D. Candidate, Rithvik Pottepalem Pharm.D. Candidate, Rachel Yang Pharm.D. CandidatePurpose:
Smoking has shown an increased risk of developing diabetes.1 However, its effects on glycemic control and HbA1c are not fully known.2,3,4,5 The study’s objective was to better determine the impact of smoking on glycemic control.
Methods:
Data was collected from the 2017-2018 National Health and Nutrition Examination Survey (NHANES) data. A composite outcome was created using the 2015 ADA criteria for glycemic control. Adequate glycemic control was classified as A1C ≤7% and fasting blood glucose within 70-130 mg/dL. Poor glycemic control was defined as participants not meeting this criteria. Pearson Chi square and Wilcoxon rank sum tests were conducted to test the significance of characteristics between smokers and non-smokers. A multivariable logistic regression (ɑ=0.05) was used to evaluate the predictive ability of the exposure and covariates for the outcome. Analyses were performed using SPSS. Inclusion criteria included those <80 years old, told by doctor they have diabetes, answered survey questions regarding smoking, and had glycohemoglobin and fasting blood glucose levels.
Results:
39.7% of the smokers versus 33.5% of the non-smokers had adequate glycemic control (p> 0.05). The results, while statistically insignificant, showed smokers having 20% lower odds of adequate glycemic control than non-smokers (95% CI 0.408, 1.583, p > 0.05). Current insulin users had 4.8 times higher odds of adequate glycemic control than non-insulin users (95% CI 2.070, 11.034, p < 0.001). Current diabetic pill takers had 2.3 times higher odds of adequate glycemic control than non-diabetic pill takers (95% CI 1.075, 4.964, p < 0.05).
Conclusions:
These findings suggest that smoking may have a negative impact on glycemic control. In the future, it may be beneficial to conduct another study with a larger population and longer follow up period.
Diabetes mellitus (DM) is the leading cause of chronic kidney disease (CKD).1 Being one of the principal causes of death worldwide,2 CKD can progress to dialysis, transplantation, and cardiovascular disease.3 Studies suggest that hyperinsulinemia enhances renal reuptake of sodium while decreasing insulin-induced vasodilation, promoting hypertension and kidney damage.4 Insulin therapy is a mainstay of DM treatment to prevent hyperglycemia by binding to insulin receptors, stimulating glucose uptake, and inhibiting glucose production.5 The study objective was to evaluate the relationship between insulin use and the presence of CKD.
The 2017-2018 National Health and Nutrition Examination Survey data was utilized. Patients were categorized into insulin-dependent or insulin-naïve. Covariates were current age, age of diabetes diagnosis, gender, smoked at least 100 cigarettes, and health insurance. The outcome of interest in the binary logistic regression was presence (CrCl<90) or absence of CKD (CrCl≥90). The multinomial logistic regression evaluated prediction of CKD stage placement compared to the reference category, stage one. Statistical analyses included Wilcoxon rank sum, Chi-squared and Fisher’s exact tests.
There were 210 insulin-dependent and 552 insulin-naïve patients. There were 116 (55.2%) patients in the exposure group and 245 (44.4%) in the control group with CrCl<90 (p=0.007). Insulin use was not significantly associated with increased odds of having CKD (aOR 1.419, 95% CI 0.937-2.150, p=.099). In the multinomial regression, insulin use was associated with increased odds of CKD stage 3 compared to CKD stage 1 (aOR 2.281, 95% CI 1.297-4.012, p=.004) and CKD stage 4+5 compared to CKD stage 1 (aOR 4.235, 95% CI 1.476-12.151, p=.007). Stages 4 and 5 were combined due to limited sample size.
Due to limited sample size and varying significance of our findings, further analysis is required to fully understand the relationship between insulin and CKD development.
Purpose:
About 116 million people in the US have hypertension1. Psychosocial factors can increase blood pressure2. The role that depression plays in the incidence of hypertension is unclear, but studies have shown a positive correlation3,4 . The PHQ-9 scoring tool used to identify symptoms and aid in a diagnosis of depression is a reliable and valid tool for analysis5. The objective of the study is to analyze the relationship between symptoms of depression based on PHQ-9 score and the incidence of hypertension.
Methods:
Data was collected using the National Health and Nutrition Examination Survey administered by the Center for Disease Control and Prevention. The Blood Pressure and Cholesterol data from 2017-2018 was used to identify patients with hypertension and patients with a PHQ-9 score ≥5. Hypertension was defined as a systolic blood pressure greater than 130 mmHg, a diastolic blood pressure greater than 80 mmHg, or taking medication for the treatment of hypertension1 .
Results:
Data was analyzed from 5,090 patients. There were 1,318 patients who reported a PHQ-9 score ≥5, with 727 having hypertension and 591 not having hypertension. There were 3,772 with a PHQ-9 score <5, with 2,120 having hypertension and 1,652 not having hypertension. Patients with a PHQ-9 score ≥5 were 4.6% less likely to have hypertension. Other factors contributing to increased incidence of hypertension were increasing age, obesity, and smoking status. Being female had a protective effect against hypertension.
Conclusions:
Patients with a PHQ-9 score ≥5 had a slightly decreased odds of having hypertension, however it can be reasonably concluded that there is no significant correlation between PHQ-9 score and the incidence of hypertension. Limitations include cross sectional data, lack of information on baseline characteristics, and incomplete past medical history. Future investigations will look into relationships between other mental health disorders and cardiovascular conditions.
Morgan PharmD. Candidate, Lauren Fortier PharmD. Candidate, Cameron Leach PharmD. Candidate, Edith Martinez PharmD. Candidate, Madison McGuire PharmD. CandidateBackground:
With obesity prevalence increasing nationally, patients are at risk for associated complications, including chronic kidney disease (CKD). Anemia commonly occurs with both obesity and CKD, though the effect of BMI on anemia prevalence in patients with CKD remains unclear.
Objective:
To evaluate the association between BMI and the prevalence of anemia in individuals with chronic kidney disease.
Methods:
This cross-sectional study utilized data from the National Health and Nutrition Exam Survey (NHANES) to evaluate 1,601 patients with CKD stages 2-5 (CrCl < 90 mL/min) between the years of 2015-2016. The exposure of interest was body mass index (BMI) and the study cohort was divided into two exposure groups; normal BMI (18.5-24.9 kg/m2) and overweight/obese BMI (>25 kg/m2). The outcome of interest was anemia, defined as a hemoglobin level < 12 g/dL in women and < 13 g/dL in men. Categorical variables were presented as frequency (%) and compared using a Pearson’s chi-square test. A logistic regression was performed, producing an odds ratio to compare exposure groups using 95% confidence intervals.
Results:
Of the 1,601 patients included in the study, 595 (37.2%) were included in the normal BMI cohort and 1,006 (62.8%) were included in the overweight/obese cohort. Comparing these two cohorts, frequency of anemia was 16% vs 14.9% (P=0.571) for normal vs overweight/obese. The adjusted odds ratio (OR) for anemia in the overweight/obese vs normal weight exposure groups was 1.603, 95% CI [0.762, 3.373] (P=0.214).
Conclusion:
Our results suggest that there is no statistically significant association between BMI and anemia prevalence in patients diagnosed with CKD stage 2 or higher.
Morgan Fenderson PharmD. Candidate, Hanna Iarussi PharmD. Candidate, Kelsey Nolan PharmD. Candidate, Mackenzie Powers PharmD. Candidate, Mariame Sylla PharmD. CandidatePurpose:
People with diabetes are twice as likely to develop cardiovascular disease and may require treatment with beta blockers (BB).1 However, many BB may worsen glycemic control.2,3 Conversely, a subset of BB may improve glycemic control.3,4 Hemoglobin A1c is a biomarker used to assess glycemic control.5 The objective of our research was to evaluate if BB have an effect on A1c in patients with diabetes compared to those not taking BB.
Methods:
Data was collected from the cross-sectional 2017-2018 National Health and Nutrition Examination Survey. The total number of participants was 806. Inclusion criteria was diagnosis of diabetes and participants lacking A1c were excluded. The exposure was BB use (n=98) and the outcome was above-goal A1c, defined as A1c ≥ 7%. An adjusted odds ratio was generated from a logistic regression adjusted for age, gender, race, BMI, anemia, and bilirubin. The outcome between exposure groups was compared using a Chi-square test.
Results:
Taking beta blockers was not associated with different A1c in patients with diabetes (aOR 1.028, 95% CI 0.667-1.585, p=0.900). Population demographics were similar between comparison groups, and there was no difference in the outcome before adjusting for covariates (p=979).
Conclusions:
This study shows that BB use does not affect A1C in patients with diabetes. Previous studies have shown worsened glycemic control with some BBs and improved control with others. It is possible these effects were lost due to our study combining these agents as one comparator. Other limitations include lack of doses, misclassification due to the limited medication list, and the inability to evaluate causality over time. Strengths include a large and diverse study population, and direct generalizability to the U.S. The clinical implication is that beta blockers may be prescribed without affecting A1c.
Cross-sectional study of the effects of prescription NSAIDs versus no prescription NSAIDs use on the Serum Creatinine of patients with arthritis.
Irina Fofanova PharmD. Candidate, Hannah Igoe PharmD. Candidate, Hyejin (Erica) Lee PharmD. Candidate, Lillian Luong PharmD. Candidate, Jasmine Takach PharmD. Candidate
Purpose:
Non-steroidal anti-inflammatory drugs (NSAIDs) used for arthritis can affect kidney function through COX inhibition and the prostaglandin pathway.1,2 This study aims to assess the effect of prescription NSAID use has on kidney function, through serum creatinine (SCr), in patients with arthritis.
Methods:
The data utilized was obtained from the 2017-2018 NHANE survey. The study included 780 participants who reported an arthritis diagnosis. Individuals who did not report a diagnosis of arthritis or had missing data were excluded. The exposure was reported use of any prescription NSAID. The outcome was elevated SCr levels, defined as a SCr greater than 1.3mg/dL for men and 1.1mg/dL for women. A logistic regression analysis was completed to assess predictive power of the exposure to the outcome.
Results:
After adjusting for the covariates age, gender, BMI, smoking status, and arthritis type, there was no statistically significant difference in the odds of elevated SCr in patients with arthritis taking prescription NSAIDs compared to those not taking prescription NSAIDs (adjusted OR = 0.695, 95% CI: 0.23 - 2.069, p = 0.384). Those aged 61 - 70 and 2'71 had statistically significant increased odds of an elevated SCr compared to those aged :S40 with adjusted OR= 5.709 (95% CI 1.303 - 25.016, P=0.021) and adjusted OR=14.374 (95% CI: 3.272 - 63.143, P= <0.001) respectively.
Conclusion: Our results were not statistically significant which correlates with results of other studies on the topic. Limitations of the study are data on indication, dose, and duration of the NSAID was not available and SCr is not a reliable measure of kidney function since variables such as dehydration and muscle mass can affect SCr. Future research should be done prospectively to be able to track kidney function over time.
Thu Le PharmD. Candidate, Aliaa Mohamed PharmD. Candidate, Elisa Piraino PharmD. Candidate, Elizabeth Shulman PharmD. Candidate, Collin Smith PharmD. Candidate
Purpose:
Diabetes mellitus (DM) and osteoporosis are two chronic conditions impacting 37.3 million and 10 million American adults, respectively 1,2 . Adults with DM are at increased risk of fractures, despite DM individuals having higher bone mineral density (BMD)3,4. However, the relationship between adults specifically with poorly controlled DM and fracture risk is not abundantly defined in current literature. Our objective was to evaluate the relationship between poorly controlled DM and fracture risk in osteoporosis patients.
Methods:
This cross-sectional study utilized data collected from the 2017-2018 National Examination Survey (NHANES) dataset. Patients 50 years and older with a diagnosis of both DM and osteoporosis were included. For the exposure, poorly-controlled DM defined as A1C > 7.0% and fasting blood glucose (BG) > 130 mg/dL compared to controlled DM A1C ≤ 7.0% and BG ≤ 130 mg/dL5. The outcome was measured by the occurrence of hip or wrist fracture4 . Multivariable logistic regression was conducted using SPSS. Chi-Square tests were used to obtain p-values for categorical data with values greater than five.
Results:
The number of poorly-controlled DM and controlled DM patients were 46 and 66 respectively. Of the poorly controlled DM group, 30 (65.2%) patients experienced the outcome of hip or wrist fractures while only 35 (53.0%) patients of the controlled group experienced the outcome (p-value = 0.199). In patients with osteoporosis, poorly controlled DM participants were 68.7% more likely (OR = 1.687) to experience fractures compared to controlled DM, however these results were statistically insignificant (p-value = 0.317).
Conclusion:
Our results suggest a statistically insignificant association between poorly-controlled diabetes and increased fracture outcomes. Additional studies with stronger causative design and larger sample size are needed to further examine whether poorly-controlled DM and fracture outcomes for those with osteoporosis are related.
This project utilizes 3D printing to assist in the investigation of the structural and mechanistic differences between the cholera toxin (CT) and the heat-labile toxin (LT). Analysis of models reveals a significant structural difference in each toxin's A2 chain, which ultimately contributes to their differential toxicities. The use of both physical and digital models facilitate a deeper understanding of these molecules.
Stroke is a leading cause of long-term disability and decreased mobility worldwide. During stroke, neuronal damage results from both the initial oxygen-glucose deprivation and the increase in excitotoxicity and oxidative stress after restoration of blood flow, a phase called Ischemia-Reperfusion Injury (IRI). Reducing the impact of these processes could result in better outcomes for post-stroke patients by preventing excessive neuronal death. Many natural compounds derived from the Cannabis Sativa plant, have been tested for neuroprotective effects due to their potential antioxidant properties. We hypothesized that neurons treated with cannabinoids immediately after experiencing an acute ischemic event will have an overall decrease in cell death compared to untreated controls. To test this hypothesis, we have performed a compound screen of various natural products on iPSC derived human cortical neurons that have been exposed to 30 minutes of oxygen and glucose deprivation. Our results suggest that neurons treated with cannabinoids following an acute ischemic event experience less cell death. These findings could provide evidence that natural products can serve as therapeutics to prevent neuronal death in post-stroke patients.
Copine 2 (CPNE2) is a poorly studied protein belonging to the “Copine" family, which are calcium-dependent, phospholipid binding proteins conserved across all species, including mammalians, where nine homologs (CPNE1-9) have been described. CPNE2 is highly expressed in the brain where we found it’s involved in the regulation of cognitive behavior, memory, and aggression in mice. Biochemical analysis of proteins interacting with CPNE2 suggests its involvement in RNA transcription, translation, and RNA stress granule formation. To understand more of the molecular function of CPNE2, we took advantage of a cell line that harbors 256 Lac operator (LacO) sequences and 96 tetracycline response elements with a minimal CMV promoter, which regulates the expression of CFP fused to the peroxisomal targeting signal SKL and 24 MS2 translational operators (MS2 repeats). Using this cell line, we investigated the role of CPNE2 in chromatin remodeling and transcription when tethered to LacO upon fusion with the Lac repressor (LacR) protein. Using live cell imaging, we investigated whether Cpne2 is recruited to the site of transcription and if calcium is needed for this process. We explored CPNE2 chromatin interaction by expressing GFP-CPNE2 in cells and used permeabilization to wash away unbound protein before microscopy analysis. Since our proteomic analysis of Cpne2 knock-out tissues revealed dysregulation of proteins involved in stress granule metabolism, we also investigated the involvement of CPNE2 in stress granule formation and clearance. We used sodium arsenite to induce stress granules in U2OS cells and determined whether RFP-CPNE2 is recruited to stress granules and if its overexpression affects stress granule clearance, using GFPG3BP1 as a marker. This study will help to understand the molecular function of CPNE2, thus elucidating its role in regulating memory and cognition, which could lead to treatments to ameliorate brain health and neurological disorders.
Astrangia poculata, the state coral of Rhode Island, is susceptible to infection by Vibrio coralliilyticus RE22, which is a common marine pathogen affecting various invertebrate species. Prior to the establishment of colonies, the settlement of coral larvae is influenced by crustose coralline algae (CCA) and their associated bacteria. Strains of Pseudoalteromonas rubra isolated from CCA have been shown to induce larval settlement of tropical corals. We hypothesize that P. rubra induces the settlement of A. poculata larvae by producing specialized metabolites packaged within membrane vesicles (MVs). In this study, we investigated MVs produced by Pseudoalteromonas rubra strains KB1 and CH007, which were isolated from CCA in the Roger Williams University Astrangia poculata coral culture tanks. MVs were isolated by ultracentrifugation, resuspended in HEPES buffer, and stored in a -80℃ freezer until further analysis. Frozen samples were extracted with organic solvents, dried on a rotary evaporator, and reconstituted in methanol. Samples were then adsorbed onto a Strata-X column, eluted with methanol and ethyl acetate, concentrated in vacuo, and reconstituted in either methanol or ethyl acetate at 1 mg/mL. Chemical analysis was completed using an LTQ-XL mass spectrometer equipped with a Phenomenex Kinetex 2.6 µm C18 (150 x 4.6 mm) LC column. We concluded that the MVs contained a family of antibacterial agents known as prodiginines. Prodiginines are a class of red-pigmented secondary metabolites that have a broad spectrum of biological activities, such as antimicrobial and algicidal activity, toxigenicity, and immunosuppressive properties. P. rubra OMVs would likely select for cargoes that exhibit antagonism towards competing strains. The future directions for this project will include further metabolomic analyses of P. rubra OMVs, with the intention of focusing on prodiginines as cargo.
Laurenobiolide
Oli Horyn, Kira Bernabe, Hannah Trautmann, Sierra Schmidt, Steven Gregory, Matthew Bertin, Kathryn M. Ramsey
Investigating the Molecular Mechanism of Action of the Sesquiterpene Lactone Laurenobiolide
Oli Horyn1, Kira Bernabe2, Hannah Trautmann2, Sierra Schmidt2, Steven Gregory2, Matthew Bertin3, Kathryn M. Ramsey2,3
1. Pharmacy Practice, University of Rhode Island, Kingston, RI
2. Cell and Molecular Biology, University of Rhode Island, Kingston, RI
3. Biomedical and Pharmaceutical Sciences, University of Rhode Island, Kingston, RI
With constantly evolving bacteria threatening the efficacy of antibiotics, the search for novel antimicrobials is imperative. Natural products historically have been used medicinally and have provided lead compounds for drug development. Laurenobiolide is a sesquiterpene lactone isolated from the North American tulip tree Liriodendron tulipifera. It has antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA). Using disc diffusion assays, we validated its activity on a methicillin-sensitive strain of S. aureus, found activity against Francisella tularensis, and observed no activity against Escherichia coli at the tested concentration. We also isolated S. aureus colonies that grew closer to the laurenobiolideimpregnated disc and confirmed that three isolates are laurenobiolide-resistant mutants. To investigate what genetic changes might lead to resistance, we used high-throughput sequencing to re-sequence the genomes of the laurenobiolide-resistant mutants and wild-type cells. This allowed us to identify what mutations are present in the laurenobiolide-resistant mutants but not the original laurenobiolide-sensitive cells. In all three resistant isolates, we found two mutations resulting in changes to coding sequences: (1) a frameshift mutation in a gene encoding a class I SAM-dependent methyltransferase leading to a premature truncation and (2) a single nucleotide change (SNP) in the rplU gene encoding the ribosomal protein bL21 that results in a change of amino acid 89 from an arginine to a proline. Given these mutations result in changes to proteins which may be essential for cellular function, we hypothesize one of these two mutations may cause laurenobiolide-resistance. Together, we validated that laurenobiolide exhibits antimicrobial activity on S. aureus, assessed laurenobiolide for antimicrobial activity against other bacteria, and identified two potentially resistance-causing mutations. Our work suggests that laurenobiolide may be developed as a novel, potentially broad-spectrum, antimicrobial and our ongoing work is expected to provide insight into how laurenobiolide exerts its effects at the molecular level.
Lung cancer is the third most common cancer that is usually diagnosed at an advanced stage and with the highest cancer-related mortalities globally, resulting in an urgent need for an effective treatment regimen upon diagnosis. Pulmonary drug delivery is most advantageous in the treatment of respiratory illness. Lipid nanovesicles are the most frequently used drug carriers due to the biocompatibility, ease of preparation and delivery of both hydrophilic and hydrophobic drugs to the targeted site. However, the dynamic environment of the lungs results in active phagocytosing of any foreign particles by the alveolar macrophages entering the respiratory tract. Infasurf® derived from calf-lung surfactant is composed of a mixture of phospholipids, neutral lipids and surfactant proteins that has composition similar to the human LS. However, their ability to form stable and effective drug carrier vesicles is not known. The present work focuses on developing and investigating physicochemical properties and in vitro therapeutic efficacy of the nano LS vesicles (LSVs). LSVs were successfully fabricated having a particle size, ζ potential and polydispersity index (PDI) of 157.3 nm, -51.4 mV and 0.269 respectively. FTIR analysis confirmed the structural integrity of the LSVs and they were found to be stable at both 4 °C and 37 °C for at least 7 days. The extruder efficiency for preparing LSVs were found to be 91.45 ± 4.40 %. Blank LSVs were found to be cytocompatible while cell uptake study displayed a lower uptake of LSVs by NR8383 as compared to the A549 cell lines. In summary, LSVs were able to form stable and effective drug carrier vesicles for pulmonary delivery of therapeutics.
The intracellular bacterial pathogen Francisella tularensis is extremely infective and can cause the potentially deadly disease tularemia. F. tularensis can survive for long periods of time in aquatic environments, creating a reservoir of bacteria that can potentially infect and cause disease in animals and humans. I found that the attenuated strain of F. tularensis, the live vaccine strain (LVS), can survive at 4°C in sterile freshwater for 21 to 56 days. We hypothesize that there are specific gene(s) that permit the survival of F. tularensis LVS at 4°C in freshwater. To identify these genes, I used a transposon insertion sequencing (Tn-Seq) approach. Specifically, I modified a transposon delivery plasmid for compatibility with a straightforward high-throughput sequencing protocol then used it to generate a transposon mutant library of ~5,500 independent mutants. To determine which mutants do and do not remain viable in freshwater over time, I inoculated the mutant library in sterile freshwater and incubated at 4°C. I collected the surviving bacteria and extracted genomic DNA at day 0, 7, and 14. I used this genomic DNA to generate Illumina sequencing libraries and had all three sequenced on a MiSeq. For each of the transposon mutant libraries, we identified ~500,000 reads corresponding to ~6,000 mutants. We identified three transposon insertion mutants in the mpl gene at days 0 and 7, but did not recover DNA corresponding to any of those insertion mutants at day 14. This suggests that mpl is important for survival in freshwater at 4°C. This gene encodes UDP-N-acetylmuramate:L-alanyl-γ-Dglutamyl-meso-diaminopimelate ligase (or Mpl). These results suggest that for maximal survival in freshwater, the Mpl enzyme must contribute to cell wall synthesis. Overall, my work demonstrates the feasibility of using this Tn-Seq technology and provides new information about how F. tularensis, an important human pathogen, is able to persist environmentally.
