Now showing items 21-40 of 12435

    • SAFE on Campus Symptom Monitoring

      Jarrell, Bruce E. (2020-08-10)
    • Pharmacokinetic Effects of -Tetrahydropalmatine on Ketamine in Rat Plasma by Ultraperformance Liquid Chromatography Tandem Mass Spectrometry

      Du, Yan; Su, Hongliang; Cao, Jie; Wei, Zhiwen; Wang, Yujin; Yun, Keming (Hindawi, 2020-07-06)
      Male Sprague-Dawley rats (n = 18) were randomly divided into three groups: a saline group (20 mL/kg by gavage), a ketamine (KET) group (100 mg/kg by gavage), and a KET (the same routes and doses) combined with levo-tetrahydropalmatine (l-THP; 40 mg/kg by gavage) group (n = 6). Blood samples were acquired at different time points after drug administration. A simple and sensitive ultraperformance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was established to determine the concentrations of KET and its metabolite, norketamine (NK), in rat plasma. Chromatographic separation was achieved using a BEH C18 column (2.1 mm × 50 mm, 1.7 μm) with chlorpheniramine maleate (Chlor-Trimeton) as an internal standard (IS). The initial mobile phase consisted of acetonitrile-water with 0.1% methanoic acid (80 : 20, v/v). The multiple reaction monitoring (MRM) modes of m/z 238.1→m/z 179.1 for KET, m/z 224.1→m/z 207.1 for NK, and m/z 275→m/z 230 for Chlor-Trimeton (IS) were utilized to conduct a quantitative analysis. Calibration curves of KET and NK in rat plasma demonstrated good linearity in the range of 2.5-500 ng/mL (r > 0.9994), and the lower limit of quantification (LLOQ) was 2.5 ng/mL for both. Moreover, the intra- and interday precision relative standard deviation (RSD) of KET and NK were less than 4.31% and 6.53%, respectively. The accuracies (relative error) of KET and NK were below -1.41% and -6.07%, respectively. The extraction recoveries of KET and NK were more than 81.23 ± 3.45% and 80.42 ± 4.57%, respectively. This sensitive, rapid, and selective UPLC-MS/MS method was successfully applied to study the pharmacokinetic effects of l-THP on KET after gastric gavage. The results demonstrated that l-THP could increase the bioavailability of KET and promote the metabolism of KET. The results showed that l-THP has pharmacokinetics effects on KET in rat plasma.
    • RSRC1 loss-of-function variants cause mild to moderate autosomal recessive intellectual disability.

      Scala, Marcello; Mojarrad, Majid; Riazuddin, Saima; Brigatti, Karlla W; Ammous, Zineb; Cohen, Julie S; Hosny, Heba; Usmani, Muhammad A; Shahzad, Mohsin; Riazuddin, Sheikh; et al. (Oxford University Press, 2020-03-31)
    • A prospective, multi-center randomized, controlled, blinded trial of vagus nerve stimulation for difficult to treat depression: A novel design for a novel treatment.

      Conway, Charles R; Olin, Bryan; Aaronson, Scott T; Sackeim, Harold A; Bunker, Mark; Kriedt, Christopher; Greco, Theresa; Broglio, Kristine; Vestrucci, Matteo; Rush, A John (Elsevier Ltd., 2020-06-19)
      Few treatment options exist for patients with difficult-to-treat depression (DTD). One potentially efficacious treatment is vagus nerve stimulation (VNS): chronic stimulation of the vagus nerve using an implanted stimulator. Given a series of recent VNS clinical studies, including a large, five-year naturalistic investigation, the Center for Medicare and Medicaid Services (CMS) reconsidered the previous non coverage determination and announced coverage for patients participating in a “coverage with evidence” trial. This study, entitled, A PRospective, Multi-cEnter, Randomized Controlled Blinded Trial DemOnstrating the Safety and Effectiveness of VNS Therapy® System as AdjunctivE Therapy Versus a No Stimulation Control in Subjects With Treatment-Resistant Depression (RECOVER), includes DTD patients with at least four unsuccessful antidepressant treatments in the current episode and will randomize both unipolar and bipolar DTD participants, each up to 500 evaluable enrollees. Predetermined interim analyses will define the necessary sample size. All participants will be implanted with VNS devices: half receive active stimulation during year one, and half receive delayed stimulation after year one. Participants will be followed for 5 years. This RCT is unique for DTD studies: 1) large sample size and long study duration (one year of controlled comparison); 2) use of a percent time in response as the primary outcome measure, given the chronic illness and its fluctuating course (vis-à-vis meeting a response criteria at a single time point); 3) inclusion of diverse measures of VNS impact on function, including quality of life, degree of disability, health status, and suicidality.
    • Atypical Skin Manifestations During Immune Checkpoint Blockage in Coronavirus Disease 2019-Infected Patients With Lung Cancer.

      Rolfo, Christian; Cardona, Andrés F; Ruiz-Patiño, Alejandro; Ariza, Santiago; Zatarain-Barron, Lucia; Pino, Luis E; Viola, Lucia; Russo, Alessandro; Rojas, Leonardo; Ricaurte, Luisa; et al. (Elsevier Ltd., 2020-07-09)
      A new coronavirus, named severe acute respiratory syndrome–coronavirus-2 by the WHO, has rapidly spread around the world since its first reported case in late December of 2019 from Wuhan, the People's Republic of China. As of mid-April 2020, this virus has affected more than 180 countries and territories, infecting more than 1,650,000 individuals and causing over 100,000 deaths. With approximately 20 million new cases globally per year, cancer affects a substantial portion of the population. Individuals affected by cancer are more susceptible to infections owing to coexisting chronic diseases (cardiovascular, pulmonary, and diabetes), overall poor health status, and systemic immunosuppressive states caused by both cancer and the anticancer treatment. As a consequence, patients with malignancies, especially those with lung cancer who develop coronavirus disease 2019, experience more difficult outcomes. A recent multicenter study carried out by the Hubei Anti-Cancer Association has also documented that patients with lung cancer had an increased risk of death, intensive care unit requirement, risk of presenting severe or critical symptoms, and use of invasive mechanical ventilation. Here, we present two representative cases of patients with lung cancer and coronavirus disease 2019 without respiratory compromise and with atypical and severe skin manifestations—findings that could be influenced by the long-term use of anti–programmed cell death protein 1 antibody.
    • Informing the role of RIFINs in malaria pathogenesis, natural immunity, and design of a severe malaria vaccine

      Zhou, Albert; Travassos, Mark A.; Laufer, Miriam K. (2020)
      Plasmodium falciparum is a eukaryotic parasite that causes severe malaria and contributed to 405,000 deaths worldwide in 2018. Victims of severe malaria are predominantly sub-Saharan African children, who typically present with symptoms of severe anemia or unarousable coma. The pathogenesis of severe malaria is poorly understood but mediated by the expression of adhesive variant surface antigens (VSAs) on infected red blood cells. VSAs are involved in sequestration and rosetting, unique virulence processes that allow the parasite to evade host immune responses and prevent clearance in the spleen. A relatively unstudied family of VSAs, the repetitive interspersed family (RIFIN) proteins, have recently been found to be important in rosetting and host immune suppression. RIFINs also appear to be targets for protective immunity; humoral immune responses against RIFINs have been correlated with asymptomatic infections. In this dissertation, I applied a multi-faceted approach using protein and peptide microarrays, transcriptomics, and reverse vaccinology to identify appealing RIFIN candidates for inclusion in a future severe malaria vaccine. I show that serological responses against epitopes within the semi-conserved domain of RIFINs associated with severe malaria reflected age-related malaria exposure. Sequencing and identifying specific rif genes expressed in clinical infections have not been feasible. I have addressed these challenges by adapting a novel bioinformatic pipeline and developing an HMM-based tool to process, assemble, classify, and subtype RIFIN sequences from peripheral blood samples. This takes advantage of a targeted probe capture method that I determined yields more abundant, full-length RIFIN sequences than other library enrichment approaches. Finally, I performed a comprehensive genomic survey of RIFIN gene repertoires using publicly available whole genome data of sixteen P. falciparum isolates to identify highly conserved, strain-transcendent sequences. Together, these results provide insights and powerful tools that can advance our understanding of the role RIFINs play in severe malaria pathogenesis and the development of naturally-acquired immunity to severe malaria. This work will aid efforts to determine targets for vaccines to protect children from the deadliest consequences of malaria.
    • Inhibition of Traumatic Brain Injury (TBI)-Induced Neuroinflammation Using Pharmacological Modulators of Metabotropic Glutamate Receptors

      Vinueza, Gelareh; Faden, A. I.; 0000-0002-1036-6378 (2020)
      Chronic dysregulated microglial activation is a major hallmark of persistent inflammation and progressive neurodegeneration following traumatic brain injury (TBI). Thus, research has focused on strategies to inhibit chronically activated microglial responses following TBI. Metabotropic glutamate receptors (mGluRs) 4 and 5 are expressed on microglia and can modulate microglial activity; therefore, they may serve as potential therapeutic targets for inhibition of microglial-dependent neuroinflammation. In the first of these studies, based on its reported neuroprotective roles, we examined the effects of the mGluR5 positive allosteric modulator (PAM) VU0360172 in an established fluid percussion injury (FPI) rat model of TBI plus hypobaria (HB). Systemic administration of VU0360172 significantly reduced pro-inflammatory cytokines, chemokines and microRNAs (miRs) at 1- and 7- days following FP+HB. However, VU0360172 did not alter injury-induced behavioral deficits examined over the following 28 days. In order to assess potential mechanisms underlying the inflammatory changes, we used Nanostring analysis to identify miRs that modulate neuroinflammation and compared plasma changes for selected miRs with brain tissue changes. The pro-inflammatory miR-223 showed the strongest correlation between plasma and brain tissue expression levels at the 7d time-point in TBI+HB experimental rodent models. An additional series of studies addressed the purported anti-inflammatory effects of mGluR4 PAMs. We employed in vitro models of immortalized microglia cell lines and primary microglia to elucidate the molecular mechanisms responsible for the modulation of inflammation by ADX88178 and other mGluR4 PAMs. ADX88178 downregulated lipopolysaccharide (LPS)-induced expression of pro-inflammatory mediators in BV2 cells and primary microglia. However, ADX88178 anti-inflammatory effects appeared to be mGluR4-independent as mGluR4 expression in our in vitro models was very low and its actions were not altered by pharmacological or molecular inhibition of mGluR4. Moreover, we showed that putative mGluR4 PAMs attenuate pro-inflammatory pathways in BV2 microglia through mGluR4/Gi-independent mechanisms involving activation of cAMP-response element binding protein (CREB) and inhibition of NFkB. Overall, these studies show that mGluR4 and mGluR5 PAMs can significantly attenuate microglial activation. Therefore, further studies should examine their potential therapeutic effectiveness after TBI.
    • Programmed Death Ligand 1 in Oral Potentially Malignant Epithelium and Implications of Regulation by Interferon Gamma

      Elnaggar, Manar; Younis, Rania H. (2020)
      Programmed death ligand 1 (PD-L1) is an immune-checkpoint regulator. Expression of PD-L1 in a subset of head and neck squamous cell carcinoma (HNSCC) was linked to improved response to immunotherapy. Oral potentially malignant disorders (OPMDs) are characterized by increased risk for malignant transformation. We investigated the expression of PD-L1 in immune cells (ICs) and epithelial cells of OPMD, which is essential to prevent progression to malignancy. Immunohistochemistry analyses in HNSCC whole excision tissue sections (104 patients) indeed demonstrated predominant expression of PD-L1 in the epithelial margins (~86%). This directly correlated with PD-L1 expression in underlying ICs (P=0.0172) with a predominating lichenoid pattern of IC infiltrate. Immunoblotting analyses demonstrated the role of human recombinant IFN-γ in the upregulation of PD-L1 expression in the oral premalignant cell line (DOK) with implications of downstream activation of pS6. Our work suggests a role for IFN-γ/PD-L1 in the immune escape of OPMDs.
    • Transient mechanical stimuli elicit rapid mechano-chemical signal transduction in non-tumorigenic and malignant mammary epithelial cells

      Pratt, Stephen JP; Martin, Stuart S.; 0000-0003-3627-7258 (2020)
      Changes in are observed during breast tumor formation and progression, which promote malignant phenotypes in both normal mammary epithelial cells and breast cancer cells. In this context, understanding the molecular details of mechanotransduction signaling may provide unique therapeutic targets. This work applied time-lapse confocal microscopy and quantitative methods to define the rapid mechanically-stimulated calcium signaling mechanisms that occur in breast epithelial cells. While most tumor cell studies focus on long-term effects of mechanical stimulation (>24h), the current approach detected an immediate initiation of cytosolic calcium signals within 2 seconds after transient mechanical stimulation. Two novel methods were developed to describe this response and underlying mechanisms: a) the real-time scratch assay and b) scratch on low elastic dishes (SLED). The real-time scratch assay revealed an ATP/P2Y2/Ca2+ signaling axis in response to scratch with implications as a path toward EMT. The second method developed was SLED, a non-damaging application of mechanical stress to breast epithelial cells with physiologic implications as the elasticity of the cell substrate better matched that of the mammary gland in vivo than the typical glass or plastic experimental dishes. New data obtained using this novel approach revealed that normal breast epithelial cells are mechanically sensitive, responding to mechanical stimuli through a two-part calcium signaling mechanism. An immediate, robust rise in intracellular calcium (within seconds) was observed followed by a persistent extracellular calcium influx (up to 30 minutes). This persistent calcium was sustained via microtubule-dependent mechano-activation of NADPH oxidase 2 (NOX2)-generated reactive oxygen species (ROS), which acted on TRPM8 channels to prolong calcium signaling. Disruption of this conserved mechanobiology mechanism was possible through oncogenic activation such that, among common oncogenic mutations, constitutively-active KRas suppressed this signaling pathway. Therefore, certain oncogenes render cells mechanically unresponsive which could have interesting implications for the evolution of cancer cell mechanosensing in the tumor microenvironment as cells acquire new mutations. In addition, altered expression of the ROS generator (NOX2) and the ROS-responsive channel (TRPM8) are indicators of reduced overall survival in ER-negative breast cancer, suggesting that this mechano-pathway identified in breast epithelial cells may also be modified in patients in vivo.
    • FOXN2 Expression: from Pluripotent Stem Cells to Neural Progenitor Cells

      Saleh, Wissam; Ament, Seth A. (2020)
      Several transcription factors (TFs) have been demonstrated as risk genes for schizophrenia (SCZ) based on genome-wide association studies (GWAS), fine-mapping, and functional studies. In this project, we prioritized TFs that have multiple lines of evidence supporting their likelihood of being causal risk loci for SCZ. We integrated results from four published studies, which used GWAS, gene expression and chromatin conformation studies to identify genes and gene networks associated with SCZ. These analyses revealed that the TF FOXN2 is a strong candidate risk gene for SCZ. Next, we characterized the dynamic expression profile of FOXN2 in our stem-cell based cortical neurogenesis model by qPCR. This study helped in establishing FOXN2 as a risk gene for SCZ potentially involved in cortical neurogenesis and building a strong base for the future genetic editing experiments designed for functional characterization of FOXN2 in cortical neurogenesis, that might relate to the pathophysiology of SCZ.
    • Delineating the Role of NKT cell Activation in B cell Lymphoma

      Lee, Michael; Webb, Tonya J. (2020)
      Natural Killer T (NKT) cells play an important role in cancer surveillance and can reduce lymphoma burden in vivo; however, a hallmark of cancer is its ability to evade immune surveillance. Our goals were to elucidate novel mechanisms utilized by B cell lymphoma to evade NKT cell-mediated immune surveillance and determine the prognostic potential of assessing NKT cell function in lymphoma patients. We found that knockdown of sphingosine kinase 1 (SK1) in human lymphoma cells results in a significant increase in CD1d-mediated NKT cell activation. Lipidomic and co-culture studies identified cardiolipin as being upregulated in SK1 knockdown cells and implicated cardiolipin as an NKT cell-specific cancer neoantigen. We also sought to determine the efficacy of NKT cell-based therapy on survival and the induction of anti-tumor immune responses in a mouse model of B cell lymphoma. We found that activation of NKT cells via early administration of α-galactosylceramide (α-GalCer) only provided modest protection. Our data suggest that the lack of protection is due, at least in part, to the expansion of myeloid-derived suppressor cells in α-GalCer-treated tumor bearing mice. Lastly, we sought to identify novel immunological biomarkers in lymphoma patients. It was found that lymphoma patients have a reduction in NKT cell function compared to healthy donors. Furthermore, lymphoma patients have significantly higher levels of both pro- and anti-inflammatory cytokines in their sera compared to healthy donors. In addition, lymphoma patients who experience relapse have significantly reduced NKT cell function in the blood, compared to lymphoma patients who did not relapse. Collectively, our studies demonstrate the multifaceted role NKT cells play in immune responses to B cell lymphoma and will help inform the next generation of cancer immunotherapy.
    • Burden and Mental Health of Family Caregivers of Cancer Patients: The Impact of Spirituality

      La, In Seo; Johantgen, Mary E. (2020)
      Background: As the primary source of care for individuals with cancer, family caregivers are relied on for treatment support and emotional care during the cancer trajectory. Studies on caregiver burden and psychological sequelae among cancer caregivers have been conducted cross-sectionally. Spirituality has been suggested as a potential buffer between burden and sequelae. Yet, there have been very few longitudinal studies addressing burden, depression, and spirituality, and there is limited information on psychometric properties of the spirituality measures in cancer caregivers. Purpose: The aims of this study were to: 1) evaluate validity of the Spiritual Perspective Scale (SPS) and explore differences in spirituality across caregiver and patient characteristics, 2) describe caregiver burden during active cancer treatment and explore caregiver and patient factors influencing caregiver burden, and 3) examine changes in caregiver burden, spirituality, and depression and explore the moderating effect of spirituality on burden-depression relationship over time. Methods: A secondary analysis of data from a longitudinal study of cancer caregivers from the NIH Clinical Center was conducted. Caregivers completed measures, including the Spiritual Perspective Scale (SPS), Caregiver Reaction Assessment (CRA), and NIH Toolbox and PROMIS® measures. Structural equation modeling and linear mixed modeling were used for testing study aims. Results: The SPS was found to have satisfactory psychometric properties in cancer caregivers. Adjusting for a direct effect of race did not alter the pattern of results, and caregivers who were older, female, racial/ethnic minorities, less educated, affiliated with a religion, and who provided care to anyone other than the patient reported higher levels of spirituality. Baseline mutuality between the caregiver and patient was negatively associated with initial burden. Changes in caregiver burden were related to being spouse caregivers, sole caregivers, and income. Scores on total burden, spirituality, and depression remained stable over time. Caregivers’ spirituality moderated the link between burden and depression (-1.26, p = .025). Conclusions: Higher levels of spirituality may act as a protective factor in the relationship between burden and depression during active cancer treatment. Identified factors related to burden and strategies to strengthen spirituality should be considered to improve caregiver mental health.
    • Dimensional Stability of CAD/CAM Patterns: A Longitudinal Study

      Byun, Shane S.; Masri, Radi, 1975- (2020)
      The goal of this study was to investigate the accuracy, over time, of computer-aided design and computer-aided manufactured (CAD/CAM) dental patterns in two different materials (resin and wax) using two different fabrication methods (subtractive and additive manufacturing). The intaglio surface of the patterns (n=48/time period) were evaluated at five predetermined time periods (zero minutes, 20 minutes, 24 hours, one week, and two weeks) relative to fabrication time. Intaglio surface scans of the generated samples were aligned with the Best Fit alignment to the design file (.stl) and compared with 3D Compare on Geomagic Control X to obtain the deviations as a root mean square (RMS). Trueness of the patterns were compared at all time points using three-way Analysis of Variance (ANOVA) (=.05). Accuracy of dental patterns deteriorated over time. When materials were considered, wax had better dimensional stability than resin. When fabrication method was considered, milled patterns had better dimensional stability than printed patterns. Time, material type, fabrication method, and all their interactions, showed a significant effect, however, the differences were very small (ranging from <1 m to 20 m). Thus, both resin and wax CAD/CAM patterns fabricated by additive and subtractive manufacturing can be used to produce dental restorations with acceptable accuracy.
    • Follicular Lymphoma Stage at Diagnosis: Determinants, Prediction from Administrative Claims Data and Impact on Healthcare Costs

      Albarmawi, Husam; Onukwugha, Eberechukwu (2020)
      Introduction: Follicular lymphoma (FL) stage is an important determinant of survival, treatment options and treatment outcomes. However, the determinants of advanced FL, defined as Ann Arbor stages III and IV, and its impact on the economic burden of FL are unknown. Moreover, for studies that rely on administrative claims data, it is not clear if advanced FL can be accurately predicted from this data source. Methods: Using the linked Surveillance, Epidemiology, and End Results-Medicare database we identified patients newly diagnosed with FL. We estimated a modified Poisson regression to explore the effect of pre-diagnosis healthcare resource utilization patterns and baseline county-level factors on FL stage. We estimated the 1-year and 5-year incremental costs of stages II-IV compared to stage I using generalized linear models. To predict FL stage from claims data, we developed and tested two random forests models. Results: We identified 11,078 patients diagnosed in 2000-2013. Half of the sample had advanced FL. Higher counts of specialist physician visits in the 3 years pre-diagnosis were associated with lower risk of advanced FL (4th quartile vs. 1st quartile: Relative Risk [RR]=0.92; 95% CI=0.86–0.99). The risk of advanced FL was 8% lower among women receiving screening mammography compared to men (RR=0.92; 95% CI=0.88–0.97). Living in counties designated as health professional shortage areas (HPSA) was associated with 7% increased risk of advanced FL (RR=1.07; 95% CI=1.00–1.14, p=0.049). In 2004-2009, FL patients with stages II, III and IV had statistically higher 1-year ($14,911; $15,106; $24,639, respectively, p<0.01) and 5-year costs ($21,590; $23,599; $34,968, respectively, p<0.01) compared to stage I patients. The random forests models exhibited poor accuracy of classifying limited and advanced FL from Medicare claims data (accuracy: ≤64%; sensitivity: ≤72%; specificity: ≤57%). Conclusions: Higher frequencies of specialist physician visits and living in counties with no HPSA can reduce the risk of presenting with advanced FL. Patients with stages II-IV incur significantly higher costs compared to stage I patients. The incremental cost increases with higher FL stage. Predicting advanced FL from claims data may not be feasible and researchers may need to rely on datasets with existing clinical information.
    • Effect of Transient Heat Exposure on Drug Delivery from Transdermal and Topical Products

      Thomas, Sherin; Stinchcomb, Audra L. (2020)
      Heating pads and electric blankets are widely used for relief from pain and to provide warmth, respectively. Their unintentional application simultaneously with a transdermal or topical system can result in unexpected drug levels in systemic circulation. Designing well-characterized in vitro and in vivo methods are vital to understanding the effect of heat and hence can aid in the development and evaluation of these products. The objective of this work was to evaluate the effect of heat on products with the same active pharmaceutical ingredient (API) but different inactive ingredients. Four drug molecules with different physicochemical properties were chosen. For each drug, formulations with different excipients were selected. In vivo serum drug profile and in vitro flux profile data can provide mechanistic understanding of heat effect on these formulations. Four topical diclofenac formulations were evaluated for heat effect in vitro under continuous heat exposure. Their flux profiles demonstrated the influence of formulation design and excipients on drug permeation at elevated skin temperature. Serum profiles of two different oxybutynin formulations evaluated under heat exposure showed very different magnitude of enhancement in serum levels under similar heat exposure conditions. Another objective of this work was establishing an in vitro - in vivo correlation (IVIVC) of heat effect on topical and transdermal formulations. This will help in characterizing and predicting heat effect minimizing the need of clinical trials and support the regulatory evaluation of these dosage forms. For buprenorphine patch, study design for in vitro permeation testing (IVPT) using human skin was well characterized to align with and mimic in vivo conditions of heat exposure. Level A and Level C IVIVC were established under normal as well as elevated temperature conditions. For lidocaine patches, IVIVC was observed for early heat effect. However, poor correlation was observed for late heat effect. The findings from this work determined IVPT studies can correlate with and be predictive of in vivo results under normal temperature conditions. But under suboptimal conditions like heat exposure, IVPT may have limitations and should be used in addition to other methods to evaluate heat effect.
    • EAP and COVID-19: Sharing the Vision: A mental Health Policy for Everyone

      Government of Ireland. Department of Health, 2020
      The publication of the Vision for Change mental health policy in 2006 set a high standard for the development of mental health policy in Ireland. As a result, there have been many important changes in the past decade aimed at improving people’s health and wellbeing. In recent times, mental health has received much attention. The outbreak of COVID 19 throughout the world created significant stress, anxiety, worry and fear for many people. The disease itself was further compounded by other impacts such as social isolation, disruption to daily life, uncertainty about employment and financial security. The Government response was rapid and a wholeof-population plan was put in place to support healthcare staff and the general population by providing health and wellbeing advice, resilience based training and providing free online interventions such as counselling and crisis texting for all in need. Indeed, the ability to create additional online interventions to augment existing services with such a wide reach, has in many ways changed how we treat the mild to moderate mental health needs of the population. In many ways the pandemic assisted Ireland to improve public attitudes to mental health because of the statutory, voluntary and community commitment to raising awareness and creating positive changes in how Ireland thinks about and delivers mental health services.
    • Post-translational Regulation of Glucokinase in Hypothalamic Neurons

      McFarland, Jennifer; Rizzo, Megan A. (2020)
      Glucose-sensing tissues utilize glucokinase (GCK), the activity of which is rate-limiting for glucose metabolism, to sense and, consequently, counteract deviations from glucose homeostasis. Post-translational regulation of GCK is well defined in the liver and the pancreas, and is critical for the maintenance of glucose homeostasis; yet, post- translational regulation of GCK in hypothalamic neurons, which play a central role in maintaining glucose homeostasis, remains relatively unexplored. Here, we use a hypothalamically-derived, glucose-sensing GT1-7 neuronal cell line to provide evidence of a receptor-driven, ER Ca2+-mediated S-nitrosylation and activation of GCK. Strategic pharmacological manipulations were paired with the assessment of GCK activity, done by either measuring NAD(P)H autofluorescence while raising extracellular glucose, or through expression of a homotransfer FRET GCK biosensor. Further, a biotin-switch assay was used to confirm the presence of GCK S-nitrosylation. This work illustrates a central mechanism of post-translational GCK regulation, which may underlie metabolic signal integration in the hypothalamus and may contribute to the pathology of diabetes.
    • A clinical evaluation of the ability of finishing files to supplement the removal of bacteria and endotoxin from primarily infected root canals (Part I- Initial evaluation)

      Kim, Eunice; Martinho, Frederico C. (2020)
      Aim: The aim of this study was to evaluate the ability of XP-endo Finisher (XPF) to supplement the removal of bacteria and endotoxin from primary infected root canals after instrumentation. Methodology: This randomized and blinded controlled trial included eight subjects. Instrumentation was performed using Vortex Blue or XP-endo Shaper, followed by supplemental instrumentation with XPF. All canals were irrigated with 2.5% sodium hypochlorite. Bacterial and endotoxin samples were taken using sterile paper points. Samples were collected before and after instrumentation and after XPF. Results: Bacteria was present in all root canals. After XPF, bacterial mean was reduced from 255 ± 311.82 CFU/mL to 2.5 ± 7.07 CFU/mL (p= .056). Endotoxin was detected in all root canals by the LAL method (KQCL test). After XPF, endotoxin mean was significantly reduced from .85 ± .26 EU/mL to .03 ± .01 EU/mL (p= .00004). Conclusion: The findings of this study showed that the supplemental use of the XP-endo Finisher after root canal instrumentation was effective in significantly reducing endotoxin but not bacteria present in primary endodontic infections.
    • Novel PC2 regulation of ezrin in renal epithelia reveals insights into ADPKD cystogenesis

      Dixon, Eryn; Woodward, Owen Maxwell (2020)
      Autosomal dominant polycystic kidney disease is caused by the loss of function of either two transmembrane proteins, polycystin-1 or polycystin-2. In renal epithelia, the consequence of polycystin loss is the formation of progressive, focal, fluid-filled cysts. However, the function and associated downstream signaling pathways specific to the polycystins have not been defined. Therefore, a new in vitro tubuloid model was designed to investigate the proximate cellular changes in renal epithelial cells following inactivation of Pkd2, the gene that encodes for polycystin-2. This model system reinforced the relevance of proteins associated with cell junctions, adhesions, and matrix in the cyst mechanism. The impact of this model was further supported through morphometrical analysis of epithelial compartmentalization in human ADPKD tissue, demonstrating an altered apical compartment in emerging cysts compared to noncystic tubules. Seeking connection between the junctions and disrupted apical compartment led to investigation of ezrin, a master scaffold in the apical compartment in renal epithelial cells. Ezrin plays a critical role in regulation of polarity, cytoskeleton organization, and protein trafficking, and the downstream consequences of its disruption have not been elucidated. Investigation into the initiating events of cystogenesis in ADPKD revealed a dramatic change in ezrin, following loss of PC2 in our tubuloid model, cystic mouse model, and pathological human ADPKD tissue. Based on this novel regulatory relationship between PC2 and ezrin, as well as the antecedent loss of ezrin to cyst formation in mice, ezrin was overexpressed in the pkd2 morpholino zebrafish model. Increased expression of ezrin diminished the formation of pronephric cysts. This lead to the design of a cyst rescue mouse model, which has exhibited promising preliminary data for cyst area reduction with additional ezrin. The disruption of ezrin in Pkd2 inducible in vitro and in vivo model systems, changes in ADPKD patient tissue, and rescue of pronephric cysts in the pkd2 MO suggest there is a role of ezrin in renal cystogenesis. Understanding the relationship of ezrin, with PC2 in renal epithelial cells will help elucidate the mechanism of ADPKD cystogenesis and define important downstream pathways necessary for epithelial functions.
    • Evaluation of the Equivalency of Generic Drugs

      Das, Sharmila; Polli, James E. (2020)
      The objective of this dissertation is to assess the bioequivalence of generic drugs. Patients with epilepsy complain about more seizures and side effects after brand-generic or generic-generic switching of an anti-epileptic drug (AED). Generic brittleness (GB) is the familiar notion that, upon switching between AEDs of pharmaceutical equivalents, a patient experiences negative outcome. Aim 1 is to probe the individual patient attributes thought to predispose a patient to generic brittleness. At the University of Maryland Medical Center, 148 patients from the outpatient epilepsy clinic were recruited for an observational case-control study. An algorithm for being GB (40% of patients) and not GB was devised. A patient with epilepsy was categorized as GB if the patient negatively opined about generics and was taking brand of their most problematic AED when generic was available. Two demographic factors that increased the odds of being GB were a patient currently taking a problem AED and increasing total number of current medications. Interestingly, taking lamotrigine increased and taking any one of six “protective” anti-epileptic drugs decreased the odds of being GB, respectively. Furthermore, no genetic, clinical laboratory or neuropsychiatry tests or their sub-elements differentiated GB patients from not GB patients. Aim 2 involves a comparative pharmacokinetic (PK) analysis upon challenging sixteen GB patients to brand-generic or generic-generic switch of an AED that they are currently on, using a four-way crossover replicate design. For each patient, test and reference PK profiles were the same, despite patients being GB. Aim 3 is to assess the noninferiority of the generic sodium ferric gluconate (SFG) against the reference product Ferrlecit with respect to drug bound iron (DBI), after single dose intravenous administration of brand and generic SFG in 44 healthy volunteers. Using a two-way crossover replicate design, plasma PK profiles of SFG to Ferrlecit were the same across two iron species (e.g. DBI and NTBI), although adverse event rates differed. In conclusion generics of AEDs and intravenous sodium ferric gluconate are bioequivalent to the brand-name drugs. Results support FDA criteria for bioequivalence in regards to AEDs and complex iron products.