Browsing by Subject "Best Poster Presentation Award"
Now showing 1 - 20 of 38
- Results Per Page
- Sort Options
Item Altered Atrial Remodeling in the Muscular Dystrophies(2019-01-22) Kappalayil, Anishka; Patel, Vishal; Cheeran, Daniel; Tassin, Tara C.; Zaha, Vlad; Peshock, Ronald M.; Mammen, Pradeep P. A.INTRODUCTION: Muscular dystrophies (MD) are genetic disorders that cause progressive peripheral skeletal myopathies. The specific mutations lead to a cycle of muscle degeneration and regeneration in MD patients, ultimately producing progressive skeletal muscle wasting. Many of the MD patients also develop associated cardiomyopathies and in 2018 is the leading cause of death. Our group has demonstrated that MD patients have very small left ventricular (LV) masses as well as depressed LVEF. This data suggest that the mode of maladaptive cardiac remodeling may be different in MD vs NICM patients. However, it remains unknown the degree of atrial remodeling that occurs in MD patients. Therefore, the central hypothesis of this study is that atrial remodeling in MD patients is altered in comparison to non-ischemic cardiomyopathy patients. METHODS: Utilizing the UTSouthwestern Cardiomyopathy Clinic, MD and NICM patients were identified. Data was extracted from cardiac MRIs to measure left atrial (LA) volumes and function. The variables used were the LA end systolic volume (LA-ESV), LA end diastolic volume (LA-EDV), and LA ejection fraction (LAEF). These measures were normalized to the body surface area (BSA). We collected data on 78 MD patients (33 MD females, 45 MD males) and 80 NICM patients (28 NICM females, 52 NICM males). Utilizing unpaired two-sided T-test, LA data was analyzed between the matched MD and NICM patients. RESULTS: The MD and NICM patient cohorts showed significant differences in the LA structure and function. CONCLUSION: Collectively, the data suggests alternative mode of maladaptive cardiac remodeling in MD vs NICM patients. Thus, further investigation into the mechanism that leads to MD-associated cardiomyopathy may ultimately identify novel therapeutic targets for the amelioration of this disease entity.Item Apolipoprotein E Isoform Influence on Outcomes after Pediatric Traumatic Brain Injury(2016-01-19) Usala, Claire; Huang, Rong; Hernandez, Ana; Miles, DarrylINTRODUCTION: The ε4 allele of the apolipoprotein E gene (APOε) is associated with poor outcomes in adults with traumatic brain injury (TBI), but its influence on recovery after pediatric TBI is uncertain. The primary aims of this study were to determine if an association exists in the outcome of children after TBI between those with at least one ε4 allele and non ε4 genotypes. Using the Glasgow outcome score (GOS), we examined three outcome variables between the two groups 1) GOS at hospital discharge, 2) GOS at long-term follow-up, and 3) the magnitude of change in GOS from discharge to > 6 month assessment (Δ GOS). METHODS: Data were prospectively collected from 220 children presenting with moderate or severe blunt head trauma between the ages of 0 and 17 years old from 2002-2013. Outcomes were assessed at hospital discharge and 12.7±8.4 months post-injury. Patients in the ε4 and non ε4 groups did not differ in injury mechanism, severity, or demographics; 23.4% had at least one ε4 allele and ε3/ ε3 was the most common genotype (67.4%). Multiple regression model analysis was conducted to determine if associations existed between the genotype combinations and outcome while controlling for age, ER GCS, ICP monitor placement, and whether CPR was performed. For ε4 genotypes analysis, we also stratified patients by admission Glasgow Coma Scale (GCS) into severe (GCS 3-8) versus non-severe (9-15), as well as moderate and severe (3-12) versus mild (13-15) groups. RESULTS: For aim 1, the GOS at discharge did not differ significantly in ε4 versus non- ε4 patients in any injury severity category before or after controlling for cofounding variables. However, after controlling for confounding variables, patients with at least one ε2 allele in the moderate or severe injury category had significantly worse GOS at discharge. For aim 2, after controlling for confounding variables, patients with the ε3/ε3 genotype had significantly better long-term GOS than patients with the genotype ε3/ε2 (p<0.05). However, we did not find a significant difference in long-term outcome between ε4 and non ε4 genotypes in the primary analysis or when stratified by injury severity groups. Finally, between ε4 and non ε4 genotypes, the Δ GOS and neuropsychological scores did not differ significantly between genotypes. DISCUSSION: Overall these results propose that unlike adults, the ε4 allele may not be associated with 12-month outcome or the rate of recovery (ΔGOS) from hospital discharge following pediatric TBI. Our results implicating worse outcomes for the ε2 genotypes suggest that this allele may be a candidate for further study to delineate its role in TBI outcome in children. Unique to this study was our analysis of neuropsychological measures, which were also not affected by the presence of ε4 in a smaller cohort of children. This study adds to current literature suggesting that unlike adults APOε4 may not exert a significant effect on pediatric TBI outcome. However, these results are limited in that any genotypic effect on neurologic repair may not be apparent for much longer time periods in pediatric brain injury as the child continues to develop and grow.Item The Association Between Tobacco Use and Intradialytic Hemodynamics in Hemodialysis Patients(2018-01-23) Sonderman, Mark; Van Buren, Peter NoelBACKGROUND: Intradialytic hypotension is known to be associated with increased mortality in maintenance hemodialysis patients. Smoking is a modifiable risk factor that is more commonly seen in patients with large decreases in intradialytic blood pressure as compared to any other intradialytic blood pressure pattern. However, the mechanisms of this association are unknown. We sought to explore the effect of lifetime tobacco use on vascular hemodynamics during dialysis. METHODS: We used impedance cardiography to measure total peripheral resistance index (TPRI) in 65 hypertensive hemodialysis patients. Additionally, we obtained blood pressure measurements before, during, and after midweek hemodialysis treatments. We then compared intradialytic hemodynamic changes between never smokers (n=35) and current or former smokers (n=30) using simple and multivariable linear regression. RESULTS: The mean change in TPRI during a single dialysis session was -438 dynes/sec/cm2/m2 in smokers and -105 dynes/sec/cm2/m2 in non-smokers (p=0.1). The intradialytic systolic blood pressure nadir was 115 mmHg in smokers and 123 mmHg in non-smokers (p=0.1). In multivariable linear regression controlling for diabetes, ultrafiltration rate, and other factors associated with intradialytic blood pressure changes, smoking was independently associated with lower nadir SBP (p=0.01) with a trend to also have a greater decrease in TPRI (p=0.08). CONCLUSIONS: Hemodialysis patients with a smoking history demonstrate a tendency towards a greater reduction in intradialytic TPRI as compared to non-smokers, with a significant independent association for lower nadir SBP. Smoking status should be aggressively ascertained in dialysis patients with significant intradialytic hypotension, but further studies are required to determine the effect of smoking cessation on intradialytic hemodynamics.Item Automated Analysis of Electroglottographic Signal in Adductor Spasmodic Dysphonia(2013-01-22) Somanath, Keerthan; Mau, TedINTRODUCTION: The human voice can be evaluated by a variety of methods. Electroglottographic (EGG) signal is produced when vocal fold vibrations produce cyclic fluctuation in the electrical impedance across the larynx. The EGG signal thus reflects the degree of contact between the vocal folds during voice production and provides a measure of voice quality based on phonatory physiology. However, the utility of EGG has been limited because existing methods of EGG signal analysis focus on the evaluation of 2-3 parameters in a segment of sustained vowel production, which does not reflect pathologies more apparent in conversational speech. We hypothesize that the EGG signal can capture perceptually relevant information from continuous speech in adductor spasmodic dysphonia (ADSD), an enigmatic speech disorder. OBJECTIVES: 1. To develop an automated computer algorithm to analyze the EGG signal in continuous dysphonic speech. 2. To identify EGG waveform features that correlate with the perceived quality of vocal strain in ADSD. METHODS: A computer program was created and refined in MATLAB to display and analyze EGG data via a graphical user interface (GUI). An automated peak-detection algorithm was developed using the differentiated EGG signal and used to perform simultaneous multi-parameter analysis on the EGG signal from normal speech and speech in patients with ADSD. Between-group comparisons were made using two-tailed Student's t test. Also, intrasubject comparison was made between strained and less-strained syllables in ADSD speech. RESULTS: A program was successfully written to allow the display and automated analysis of EGG data from samples of continuous dysphonic speech. The program was found to generate data with good internal consistency. Application to normal and ADSD subjects showed that the open quotient parameter was able to distinguish between strained and less-strained syllables with statistical significance (p=0.04). DISCUSSION/CONCLUSION: We have developed a method to analyze EGG signal from samples of continuous dysphonic speech. The numerical and graphical data obtained support the utility of EGG as an objective means to clinically highlight the speech differences between normal subjects and subjects with ADSD. Further testing to establish normative values for the analyzed EGG parameters and their subsequent comparison with patient EGG data is required to affirm their utility for routine clinical voice assessment.Item A Bacterial Cholesterol Sensor to Assess Cholesterol Accessibility in Red Blood Cells(2016-01-19) Chakrabarti, Rima Shah; Radhakrishnan, Arun; Cohen, Jonathan C.; Hobbs, Helen H.Mammals are able to gain cholesterol from two sources: diet and endogenous synthesis. However, the only means of cholesterol removal is reverse cholesterol transport (RCT), in which cholesterol is transported to the liver and exported into bile. While high density lipoprotein (HDL) is considered to be the major conduit for RCT, studies with HDL-deficient animals reveal no defect in tissue cholesterol balance. We hypothesize that red blood cells (RBCs), which contain 50% of blood cholesterol, also play a role in RCT. To measure accessible cholesterol in RBCs, we developed an assay that utilizes the cholesterol binding properties of the toxin Anthrolysin-O (ALO). We purified and fluorescently labeled domain 4 of ALO (fALOD4). We then incubated fALOD4 with RBCs from 164 subjects and measured fluorescence intensity using flow cytometry. Both intra-assay and intra-individual variability of the assay were less than 10%. In the test population, fALOD4 binding varied 10-fold. fALOD4 binding did not correlate with total RBC cholesterol but did correlate with RBC phosphatidylcholine (PC) (-0.42, p=6e-7) and lyso-phosphatidylcholine (LPC) (0.40, p=6e-6). Increasing the LPC:PC ratio in RBCs with phospholipase A2 (PLA2) increased fALOD4 binding by 3-fold. fALOD4 binding also correlated with plasma HDL (0.30, p=6e-4) and triglycerides (-0.57, p=2e-12). These data suggest that RBC accessible cholesterol varies in a population, is driven by intrinsic RBC phospholipid composition and interacts with known cholesterol transporters in the blood. Future studies will determine if variability in fALOD4 binding is driven by non-lipid RBC membrane components, is genetically determined, or contributes to atherosclerosis.Item Calcitriol Treatment Suppresses Contraction-Associated Gene Expression in Pregnant Mice Near Term(2018-01-23) Morgan, Kelsi; Yang, Ailling; Montalbano, Alina P.; Mendelson, Carole R.Preterm birth (PTB) is the leading cause of infant mortality during the first four weeks of life world-wide. This is due, in part, to our incomplete understanding of the mechanisms that mediate uterine quiescence during most of pregnancy and promote the transition to labor at term. Term and preterm labor are associated with increased levels of proinflammatory cytokines within maternal reproductive tissues where they activate inflammatory transcription factors (e.g. NF-κB) that enhance expression of genes encoding contraction-associated proteins (CAP) (i.e. connexin-43 (CX-43)), oxytocin receptor (OXTR)). By contrast, uterine quiescence is maintained throughout most of pregnancy by increased progesterone (P4) levels and enhanced progesterone receptor (PR) activity, which silence expression of proinflammatory mediators and CAP genes. Treatment of pregnant women at risk for preterm labor with progestins has negligible effects - underscoring the need for novel therapeutic targets. To identify such targets, our lab surveyed the myometrial transcriptome of timed-pregnant mice at 15.5-18.5 days post-coitum (dpc) and during labor at term (19.0 dpc) using RNA-sequencing. Interestingly, Cyp27b1 was one of the most highly downregulated transcripts at 18.5 dpc and in-labor, compared to 15.5 dpc. Cyp27b1 encodes 1α-hydroxylase, the key enzyme responsible for synthesis of the active form of vitamin D, 1,25-dihydroxyvitamin D3 (calcitriol) which binds to the vitamin D receptor (VDR). Calcitriol/VDR have anti-inflammatory actions and are reported to mediate maternal tolerance to the hemi-allogeneic fetus. Interestingly, we previously observed that P4 treatment of timed-pregnant mice caused a significant increase in myometrial CYP27B1 mRNA levels, compared to controls. In the present study, we sought to assess effects of calcitriol treatment on CAP gene expression in timed-pregnant mice. To this end, pregnant mice were injected s.c. daily from 13.5-17.5 dpc with vehicle (n=3) of with 3 μg/kg of calcitriol (n=4). Mice were sacrificed and myometrial tissues were collected at 18.5 dpc. RT-qPCR revealed significantly reduced levels of CX43 (p<0.0001) and OXTR (p<0.05) in myometrium of calcitriol treated mice, compared to controls. Collectively, these data suggest that the decrease in Cyp27b1 expression, coupled with the decline in PR function near term may contribute to increased CAP gene expression leading to myometrial contractility and labor. Cyp27b1 may serve as a key P4/PR target gene that acts cooperatively to maintain myometrial quiescence via its anti-inflammatory actions. Thus, calcitriol may be a safe and effective treatment for the prevention of PTB.Item Characterization of β-glucuronidase for Enzyme Replacement Therapy in DYT6 Dystonia(2023-01-31) Lieu, Linh; Yim, Daron; Pappas, Samuel; Dauer, WilliamBACKGROUND: Dystonia is a debilitating disorder defined by sustained involuntary twisting movements. The current symptomatic treatments for dystonia offer only modest efficacy but numerous side effects. Dominantly inherited, loss of function mutations in the THAP1 transcription factor cause DYT6 dystonia (DYT-THAP1). THAP1 modulates the development of oligodendrocyte progenitor cells (OPC) by regulating the catabolism of glycosaminoglycans (GAGs), a crucial component of the extracellular matrix. The loss of THAP1 within OPCs directly reduces GAG-catabolic lysosomal enzyme β-glucuronidase (GusB) causing the accumulation of GAGs that inhibit their own maturation to myelinating cells. The result is severe dysmyelination during early CNS maturation and impaired neurodevelopment. Genetic overexpression of GusB rescues the maturation deficits and CNS myelination in THAP1 deficient mice raising the critical question of whether β-glucuronidase enzyme replacement could restore myelination in THAP1 null mice. OBJECTIVE: Characterization of β-glucuronidase for enzyme replacement therapy (ERT) in DYT6 dystonia models. METHOD: To establish a symptomatic model of DYT6 dystonia, we utilized a Cre/LoxP based Thap1 conditional knockout mouse model ("THAP1-NCKO") and a GusB transgenic mouse line ("GusB-TG"). We evaluated the enzymatic activity and biodistribution of GusB in the CNS using biochemical and histochemical assays. RESULTS: We determined that THAP1-NCKO mice had lower GusB activity than their control counterparts. Interestingly, the activity of GusB is higher in adult (P60) mice compared to juvenile (P30) mice. Visualization of GusB activity showed that distribution of GusB was highest in white matter tracts. We showed that mice with THAP1- related deficits experienced a significant reduction in GusB activity within white matter tracts but not in other surveyed GusB positive brain areas. CONCLUSIONS: Age differentially affects CNS GusB enzymatic activity in a murine model of DYT6 dystonia. GusB enzyme exhibits a distinct biodistribution that varies regionally. White matter tracts experience more severe defects with THAP1 loss. Our results provide insights into the specific locations where GusB activity is deficient and highlight the importance of a "critical period" in which genetic insults have long lasting neurodevelopmental implications.Item Comparisons of Anterior Vaginal Wall Indentation Parameters in Age-Matched Control and Prolapse Patients Using an Operator Independent Artificial Finger(2018-01-23) Wang, Connie; Abraham, Michael R.; Abrego, Christopher E.; Shiakolas, Panos S.; Zimmern, Philippe E.GOAL: To compare reaction forces of the human anterior vaginal wall in control (C) and prolapsed (P) women in response to pressure applied at different angles of indentation through an automated artificial finger equipped with a distal sensor. METHODS: Following IRB approval, a tripod-mounted, artificial finger equipped with a calibrated, piezoresistive sensor at its tip and automated by NI LabView 2015 software for motion control via an actuator was used to create anterior vaginal wall deformations at 10, 15 and 20 degree angles. Age-matched women in the C and P groups were compared. All measurements were performed in the supine position in the operating room, with patients under general anesthesia prior to the start of the operation and after the bladder was drained. Each deformation included a 1 second upwards indentation, a 1 second maintenance "hold", and a 1 second return of the fingertip to the baseline. Measurements were done in triplicate with a 3 second interval between each deformation sequence. Real-time voltages, equivalent to reaction forces sensed by the sensor during each indentation, were modeled as function of motion profiles and analyzed in Excel. The motion profile of each indentation was used to calculate baseline voltage, amplitude change over the 1 second interval of upwards indentation, and slope of the upwards indentation curve in its median 0.5 second range. RESULTS: Five women of similar age group (mean 64, 51-73) were studied in each group. A significant difference was observed between all degrees of indentation in baseline voltage in P and C groups (p<0.05). At 10 and 20 degrees of indentation, there was a significant difference in amplitude change between P and C groups, while there was a significant difference in slope of indentation at 15 degrees between P and C groups. CONCLUSION: The biomechanical properties of the human anterior vaginal wall can be objectively determined by a new device resembling the human finger. This mounted, free-standing artificial finger can apply a predictable and reproducible deformation to the anterior vaginal wall to compare the indentation properties of vaginal tissue in prolapsed and non-prolapsed conditions.Item Corneal Stromal Remodeling after Photorefractive Keratectomy(2017-01-17) Su, Shan; Kivanany, Pouriska; Grose, Kyle; Petroll, W. MatthewThe cornea is an optically clear tissue that contributes 2/3 of the eye's refractive power, making it a target for vision correction procedures. A small percentage of patients who receive corneal surgery experience loss of corneal transparency (haze). Haze occurs when stromal cells in the cornea (keratocytes) become activated and transform into fibroblasts or myofibroblasts, which can generate contractile forces that may disrupt the collagen architecture. We investigated the wound healing process following photorefractive keratectomy (PRK, a clinical vision correction procedure) using 3-D imaging both in vivo and in situ. We hypothesized that following PRK in rabbits, keratocytes located within the injured stroma, where collagen is intact, would align with the collagen lamellae and will not produce fibrosis. In contrast, we expected keratocytes anterior to the wound, where collagen is disrupted, to undergo myofibroblast transformation and produce significant haze. Twelve New Zealand white rabbits were scanned one week before surgery using an in vivo confocal microscope. PRK was performed on the right eyes of the rabbits (9 diopter spherical photocorrection) with subsequent scans at 7 days, 21 days, 3 months, and 6 months. Custom software was used to build 3-D reconstructions and measure stromal thickness and haze. Some rabbits were sacrificed at each time point to obtain in situ confocal images. Thickness measurements at pre-scan, 7 days, 21 days, 3 months, and 6 months were: 335, 208, 265, 293, 313um. The decrease and subsequent increase in thickness is consistent with removal of tissue for PRK, followed by tissue regeneration. Haze measurements at the same time points were: 1797, 4453, 6906, 3212, 3433 intensity units. The increase and subsequent decrease in backscatter suggests two phases of wound healing. In situ confocal imaging showed that cells within the native stroma were aligned with collagen lamellae without prominent stress fibers at 7 and 21 days, while cells anterior to the injured stroma at 21 days aligned randomly and displayed prominent stress fibers. At 3 months, these cells aligned with correlation to the collagen and did not express stress fibers. At 6 months, the cell/collagen arrangement was similar to uninjured tissue. Our results suggest that the collagen lamellae direct fibroblast patterning during repopulation of the native stroma, without inducing fibrosis or significant haze. In contrast, cells accumulating on top of the stroma initially align randomly and produce hazy, fibrotic tissue. Remarkably, over time, cells remodel the fibrotic tissue to produce a lamellar structure that is similar to the native corneal stroma.Item Developing a Real-Time, Axially Resolving Optical Monitor of Spinal Cord Blood Flow(2019-01-22) Gao, Feng; Busch, David R.; Goh, Chia Chieh; Lin, Wei; Yodh, Arjun G.; Floyd, Thomas F.BACKGROUND: Spinal cord ischemia is a disease of high morbidity and mortality often caused by surgeries repairing the thoracic and abdominal aorta. Current methods to monitor spinal cord hemodynamics such as electrophysiology methods, MR arterial spin labeling, and laser Doppler either have a slow response time or are unfeasible intra-operatively. In this study, we developed an optical probe to monitor spinal cord blood flow and oxygenation in real-time at multiple sites along the spine. METHODS: Experiments were conducted on 8 adult domestic pigs. Probes were inserted into the epidural space through a laminotomy prior to asphyxia and local ischemia via catheter balloon inflation. Vital signs, anesthetic parameters, and spinal hemodynamics were measured continuously prior to intervention, throughout asphyxia, and during inflation/deflation of the balloon catheter. Optical blood flow measurements were compared against microspheres. Optical hemoglobin saturation of spinal cord was compared to mixed venous blood gases. RESULTS: The fiber optic probe detected changes in flow and oxygenation in all asphyxia and balloon inflation trials across multiple sites along the spine. We observed significant changes in spinal cord blood flow during balloon inflation in the epidural space. We also observed a significant correlation between optically measured hemoglobin saturation and mixed venous blood gases. CONCLUSION: We developed an intra-operative tool that provides continuous, real-time monitoring of spinal cord hemodynamics at multiple sites along the spine. We hope this tool can more safely guide surgeons in reducing the incidence of spinal cord ischemia.Item Development of Deep Learning Artificial Intelligence to Detect Osteoporosis(2024-01-30) Fan, Christopher; Scanio, Angelo; Joshi, Parag; Öz, Orhan K.; Peshock, Ronald M.; Kay, FernandoOsteoporosis poses a substantial social and economic burden, with estimated treatment costs reaching a combined six trillion USD in the USA, Canada, and Europe. Although dual-energy X-ray absorptiometry (DEXA) is the diagnostic gold standard, computed tomography (CT) scans have proven to be reliable proxies for bone density measurement. Opportunistic screening for low bone density using CT obtained for other purposes can potentially reduce complications from osteoporotic fractures and health care costs. In this study, we developed an artificial intelligence (AI) algorithm using neural networks and the MONAI library to estimate DEXA bone density from non-contrast cardiac CT obtained for coronary calcium scoring purposes. A total of 2797 Dallas Heart Study phase 2 participants (39% male, 61% female) were included. The AI algorithm was first developed to automatically segment trabecular bone from cortical bone. This was trained and validated with manual segmentation of the trabecular bone by two medical students, a radiologist, using MONAI 3D autoseg. The ML algorithm achieved a Dice score of 0.97 when compared to human segmentation. A second AI model was developed utilizing segmentations of the first model. This AI was trained utilizing corresponding DEXA bone mineral density (BMD) for thoracic vertebrae. The best performing model was trained for 102 epochs, resulting in a training root mean square error (RMSE) of 0.0628 mg/cm2 and validation RMSE of 0.0842 mg/cm2. The final AI algorithm predictions yielded an R2 value of 0.71 compared to DEXA (Figure 1). Our findings underscore the clinical feasibility for an automated neural network to predict DEXA scores from non-contrast cardiac CT. This approach may help in the early detection of unsuspected low bone mineral density in patients undergoing CT scans for other reasons, allowing for potential improvements in patient outcomes and resourceful utilization of diagnostic imaging.Item Effect of ULK1 Inhibition on Corneal Epithelial Cells During Pseudomonas aeruginosa Infection(2024-01-30) Abdallah, Joelle; Ayilam Ramachandran, Rajalakshmy; Robertson, Danielle M.INTRODUCTION: Pseudomonas aeruginosa (PA) keratitis is a severe infection of the cornea that can lead to blindness. Studies in our lab have shown that PA exploits autophagy, a major cellular degradation process, in corneal epithelial cells (hTCEpi cells) to promote intracellular survival. We have further shown that the inhibition of autophagy by the Unc 51-like kinase (Ulk1), an enzyme that mediates formation of the autophagosome, reduces intracellular levels of PA. More recently, we have demonstrated that PA infection negatively impacts host mitochondria. ULK1/2 has been reported to translocate to mitochondria to mediate mitophagy however, a role for ULK1/2 in mitochondrial homeostasis during infection has not yet been explored. In this study, we investigated the effects of the inhibition of Ulk1 during PA infection on host mitochondria. METHODS: Telomerase-immortalized human corneal epithelial (hTCEpi) cells were used for this study. Cells were cultured in serum-free defined keratinocyte media with growth supplements. Cells were inoculated with 106 CFU/ml of PA in log growth phase with or without treatment with 1 ï�M of the Ulk1/2 inhibitor MRT68921. Intracellular levels of PA were quantified using a gentamicin survival assay. Oxygen consumption and mitochondrial polarization were assessed using Seahorse metabolic flux analysis and tetraethyl-benzimidazolyl-carbocyanine iodide (JC-1), respectively. Levels of pro-inflammatory cytokines were assessed using ELISA. Untargeted metabolomics was performed using mass spectrometry. Cellular changes were further evaluated using transmission electron microscopy (TEM). RESULTS: PA infection induced robust mitochondrial depolarization. There was a corresponding increase in secretion of IL-6 and IL-8. Treatment with MRT68921 restored mitochondrial polarization and reduced IL-6, but had no effect on IL-8. MRT68921 also reduced intracellular levels of PA. TEM demonstrated robust mitochondrial fission during PA infection. Treatment with MRT68921 preserved mitochondrial structure and polarization during PA infection. CONCLUSIONS: Taken together, these data suggest that Ulk1/2 modulates the host mitochondrial response to PA infection. Further studies are needed to determine the mechanism by which MRT68921 preserves mitochondrial function and its potential use as an adjunct therapeutic for PA-mediated keratitis.Item Employing Alternating Magnetic Fields for Biofilm Destruction(2020-01-21) Vachon, M. Jonathan; Wang, Qi; Pybus, Christine; Shaikh, Sumbul; Chopra, Rajiv; Greenberg, DavidBACKGROUND: Prosthetic Joint Infections (PJI) are a common complication of implant surgery. Due to biofilm formation, treatment is costly, includes weeks of antibiotic therapy, and even total replacement of the prosthesis. However, a non-invasive thermal method of biofilm eradication has recently been developed: using high-frequency alternating magnetic fields (AMF) to destroy biofilm via induction. METHODS: The experiment used Staphylococcus aureus, a prototypic pathogen implicated in PJI. Stainless steel rings were used to mimic prosthetic joints. Biofilms of Methicillin-Sensitive Staphylococcus aureus (UAMS1-lux) were grown on stainless steel rings in a shaking incubator for 24 hours, 110rpm at 37°C, in Tryptic Soy Broth media. The rings were then resuspended in fresh media and incubated for another 24 hours at 110rpm, 37°C. Untreated rings, Antibiotics only, AMF only, and a combination of AMF + Abx were tested. Biofilms in the latter 2 categories underwent 3s pulses of AMF exposure every 5 minutes for 15, 30, or 60 minutes at a target temperature of 65°C every 12 hours for 24 hours. Ceftriaxone (2.0μg/ml) was used for the Abx conditions. Rings were sonicated at the indicated timepoints and colony-forming units (CFU) were determined. RESULTS: A synergistic effect between AMF and antibiotics was seen. At 12 hours, the Abx only and AMF only treatments showed regrowth; however, the combination therapy showed a 2.1-log decrease in biofilm CFU. Similarly, at 24 hours, solo AMF treatment showed total regrowth and Abx only treatment showed modest bactericidal effects (2.1 log reduction). However, combination therapy at 24hr showed a 5.35 log reduction and reached the limit of detection of the assay. Additionally, we are investigating the effects of AMF with Linezolid (2.0 μg/ml). At 24hrs, a 4.3 log reduction in biofilm CFU was observed in the combination treatment, while solo treatments showed total regrowth. CONCLUSIONS: These in vitro results serve as a strong basis for future work on AMF utilization in treatment of PJI. AMF and antibiotics are synergistic in reducing biofilm off metal. The observed bactericidal effects combined with this non-invasive means have wide and significant implications in improving the patient's quality of life as well as improving healthcare costs of PJI treatment.Item Gamification of Exercise and Its Application for Fall Prevention Among Patients with Diabetes and Peripheral Neuropathy(2014-02-04) Monier, Elizabeth; Najafi, Bijan; Grewal, Gurtej; Lee-Eng, Jackie; Menzies, Robert; Talal, Talal K.; Armstrong, David G.BACKGROUND: Individuals with diabetic peripheral neuropathy (DPN) often experience concomitant impaired proprioception and postural instability. Diminished peripheral sensory input, as found in DPN, has been associated with an increased risk of falling in elderly diabetics. Conventional balance training often consisting of Tai Chi, physiotherapy, and strength training has demonstrated an improvement in effective balance control for DPN patients, but these conditioning regimens do not provide visual feedback to help compensate for impaired proprioception. METHODS: In a randomized trial study, the efficacy of an innovative game-based balance and proprioception training program for patients with DPN was assessed through direct evaluation of changes in body sway before and after the exercise protocol using body worn sensor technology. Participants were randomized to either intervention or sham groupings. All participants' baseline gait and balance were assessed at the initial visit and again after four weeks. Twice weekly, the intervention group participated in a training regimen based on virtual simulation for a total of four weeks. Postural sway was assessed before and after each training session. The gaming exercise consisted of a series of ankle point-to-point reaching tasks as well as crossing a series of virtual obstacles of varying heights. During exercise training, the body-mounted sensors connected to the created program to produce real-time animation of lower extremity joint position for the participant to view on a computer monitor. RESULTS: Forty-one eligible subjects have been recruited to date; however, the results of 15 participants (Age: 56.3 ± 4.9, BMI: 30 ± 15 kg/m2) who completed the four-week exercise program have been reported. The preliminary results suggest that the active group reduced ankle sway by 76% (2.82 ± 2.8° to 0.66 ± 0.47°), hip sway by 81% (7.96 ± 9° to 1.48 ± 1.2°) and center of mass (COM) sway by 76% (0.69 ± 0.7° to 0.16 ± 0.11°) during eyes open balance assessment. Similar reductions during eyes closed assessments were observed with reductions of 50%, 24% and 45% for ankle, hip and COM sway, respectively. CONCLUSION: This research implemented a novel balance rehabilitation strategy for patients with diabetic peripheral neuropathy based on virtual reality technology that helps compensate for impaired joint proprioception. The method employed body sensors to generate an interactive user interface for real-time visual feedback based on ankle-joint motion, similar to a video game. The study provides evidence that visual illustration of extremity position in an interactive setting coupled with motor control tasks may be an effective rehabilitation method for postural instability in patients with diabetic peripheral neuropathy.Item Genus-Level Identification of Bladder-Resident Bacteria Associated with Recurrent Urinary Tract Infection in Post-Menopausal Women by Fluorescence In-Situ Hybridization(2022-02-01) Kenee, Parker R.M.; Gadhvi, Jashkaran G.; Khan, Fatima; Christie, Alana; De Nisco, Nicole; Zimmern, Philippe E.INTRODUCTION/BACKGROUND: Antibiotic-recalcitrant recurrent urinary tract infection (rUTI) is common in postmenopausal women. Many women elect to undergo electrofulguration (EF) of areas of chronic cystitis when standard antibiotic therapies fail. One potential benefit of this procedure is the removal of tissue-resident bacterial communities.1 Although tissue-resident bacteria have been observed in the bladder walls of postmenopausal women with rUTI by fluorescence in situ hybridization (FISH), genus and species level identification of these bacteria by FISH has not yet been reported.2 Here, we use genus-specific probes to quantify Escherichia spp. present in the bladder biopsies taken from postmenopausal women with rUTI undergoing EF and investigate the relationship between detected bacterial community sizes and stage of cystitis. METHODS: Following IRB approval, bladder biopsies were obtained from consenting postmenopausal women who elected EF for the advanced management of rUTI. Biopsies were immediately fixed in paraformaldehyde and then paraffin-embedded and sectioned (5 μm). FISH was performed as described previously2 using the following probe sets: 1. Scramble-AlexaFluor488/647 (control), 2. Universal 16s rRNA-AlexaFluor647 (all bacteria), and 3. Escherichia 16s rRNA-AlexaFluor488. Slides were imaged using a Zeiss LSM880 with a 63x objective. 10 randomly sampled images were collected for each biopsy section. Images analysis was performed blinded to quantify bacterial community sizes. Patients were classified by cystitis stage (Stage 1 (trigone alone) to 4 (pancystitis)) determined by cystoscopy at time of the biopsy. Least mean squares statistical analysis was used to generate an average number of bacterial organisms per 10x1 μm2 for each cystitis stage. RESULTS: Genera-specific FISH was performed on bladder biopsies from 23 women. The universal 16s rRNA probe detected tissue-resident bacteria in the biopsies of 95.7% (22/23) women. Tissue-resident Escherichia spp. were detected in the bladder biopsies of 52.1% (12/23) women. The highest average bacterial community sizes were observed in Stage 1 cystitis bladder biopsies (8.4 per 10x1 μm2, 95% C.I. 6.6 - 10.1). CONCLUSION: For the first time, 16s rRNA FISH was used to detect Escherichia spp. in the bladders of postmenopausal women electing to undergo EF for the advanced management of rUTI. Interestingly, bladder-resident bacterial community sizes were highest in bladder biopsies from women with Stage 1 cystitis.Item Identification of Drivers of Tumor Lymphangiogenesis in Non-Small Cell Lung Carcinoma (NSCLC)(2014-02-04) Sibley, Robert Carson; Dellinger, Michael; Brekken, RolfBACKGROUND: Non-small Cell Lung Carcinomas (NSCLCs) frequently spread to regional lymph nodes before they colonize other regions of the body, and the status of regional lymph nodes is an important prognostic factor for predicting the outcome of patients with lung cancer. It has recently been demonstrated that lymphangiogenesis, the sprouting of new lymphatic vessels from pre-existing vessels, facilitates the lymphogenous dissemination of NSCLC. However, the molecular mechanisms driving lymphangiogenesis in NSCLC are poorly understood. Objective: Our aim was to identify novel lymphangiogenic genes by identifying lymphangiogenic lung tumor cell lines, and then to use microarray data to generate a "lymphangiogenic" gene signature. METHODS: Tumors from 13 lung tumor cell lines were stained with antibodies against LYVE-1 and Podoplanin. Lymphatic vessels were counted in 5 representative 20X fields per tumor. Average lymphatic vessel densities were then calculated. Cell lines were grouped into lymphangiogenic, non-lymphangiogenic, and intermediate categories. Microarray data from the two extreme groups were then compared to generate a "lymphangiogenic" signature. RESULTS: Four cell lines, (Calu-1, H1993, HCC461, and HCC827) displayed high intratumoral lymphatic density, and five cell lines (Calu-3, H1155, H1395, H1975, and H2073) displayed no intratumoral lymphatic vessels. The "lymphangiogenic" signature obtained from the microarray data from these groups contained 146 genes, including the lymphatic growth factor VEGF-C. CONCLUSIONS: Our preliminary findings suggest that VEGF-C is an important driver of tumor lymphangiogenesis in NSCLC. The other 145 genes in the signature may also serve novel functions in regulating tumor lymphangiogenesis. Together, the results from this project provide mechanistic insight into the process of tumor lymphangiogenesis and metastasis. We believe that this information will lead to the development of new prognostic or predictive markers and therapeutic strategies to improve the outcome of patients with lung cancer.Item Increased Amyloid Deposition after TBI Correlates with Cognitive Deficits and Symptom Worsening(2016-01-19) Sherman, Matt; Gatson, Joshua; Stebbins, Cari; Kilianski, Joseph; Madden, Christopher; Wolf, Steven E.; Diaz-Arrastia, Ramon; Batjer, Hunt; Minei, JosephBACKGROUND: Traumatic brain injury (TBI) is a risk factor for Alzheimer's disease (AD). The primary objective of this case-series study was to conduct early 18F-AV-45 (florbetapir F18) positron emission tomography (PET) imaging in mild-to-moderate TBI subjects after injury to determine if amyloid plaque load predicts cognitive deficits. METHODS: Serial florbetapir F18 PET imaging was conducted in 7 individuals with a mild or moderate TBI (as indicated by their Glasgow Coma Scale [GCS] score between 13 and 15) at day 14 and at 12 months after injury. Of the 7 subjects that were tested, only one had a moderate TBI. Amyloid plaque levels were measured in the cerebral cortex of each individual. To screen for cognitive deficits, the symbol match test was administered at 12 months after TBI. RESULTS: At day 14 after injury, compared to healthy controls, the mild and moderate TBI subjects (N=7) had a 10% increase in amyloid plaque load within the cerebral cortex. When stratified by cognitive outcomes, at day 14 after injury, the subjects with poor outcomes (n=3) experienced a 20% and 13% increase in brain amyloid compared to healthy controls and TBI subjects with good outcomes, respectively. With respect to cognition, at 12 months after injury, the subjects with poor outcomes exhibited a negative correlation (r= -0.71) between amyloid plaque load and cognitive performance. Also, a positive correlation (r=+0.78) was detected between increased brain amyloid load and symptom scores at 12 months. CONCLUSIONS: Individuals with early, substantial increases in brain amyloid experience poor outcomes in the form of memory dysfunction and heightened symptoms (memory deficits, headaches, difficulty concentrating, etc.) at 12 months post-injury. Data presented here suggests that florbetapir F18 PET imaging may be a sensitive biomarker for predicting outcomes within the mild and moderate TBI population.Item Intravascular Ultrasonography Analysis of the Everolimus-Eluting Stent in Coronary Chronic Total Occlusions(2014-02-04) Navara, Rachita; Michael, Tesfaldet; Papayannis, Aristotelis; Patel, Vishal; Fuh, Eric; Alomar, Mohammed; Moin, Danyaal; Brayton, Kimberly; Mogabgab, Owen; Shorrock, Deborah; Tran, Daniel; Roesle, Michele; Rangan, Bavana; Haagen, Donald; Makke, Loren; Abdullah, Shuaib; Luna, Michael; Addo, Tayo; Banerjee, Subhash; Brilakis, Emmanouil S.PURPOSE: Chronic total coronary occlusions (CTOs) are challenging to treat in part due to high rates of restenosis after stenting. Drug-eluting stents improve outcomes compared to bare metal stents. The goal of the present study was to evaluate the angiographic, intravascular ultrasonography (IVUS) and clinical outcomes after implantation of the Everolimus-Eluting Stent (EES) in CTOs. METHODS: One hundred consecutive CTO patients who were successfully treated using EES at the Dallas VAMC between 2009-2012 were enrolled in the AngiographiC Evaluation of the Everolimus-Eluting Stent in Chronic Total Occlusions (ACE-CTO trial: NCT01012869). Patients underwent follow-up angiography and IVUS imaging at 8 months and clinical follow-up at 12 months. The primary endpoint of this study, binary angiographic restenosis, was defined as >50% minimum lumen diameter stenosis at 8-month follow-up quantitative coronary angiography. The primary endpoint of the IVUS analysis was 8-month in-stent neointimal hyperplasia (NIH) volume (stent volume-lumen volume). RESULTS: Patients had high prevalence of hypertension (91%), hyperlipidemia (90%), diabetes (47%), prior MI (51%), and prior PCI (21%). Of the 89 patients who underwent follow-up angiography, binary in-stent angiographic restenosis occurred in 41 patients (46%), and IVUS analysis was performed in 61 patients. IVUS was not performed in 24 patients (8 of whom had occlusive in-stent restenosis), and suboptimal image quality precluded analysis in 4 patients. Mean and median neointimal hyperplasia volume were 68 ±100 and 26 (0, 91) mm3, respectively. This corresponded to a mean and median percent volume obstruction of 12% ± 15% and 5% (0%, 24%), respectively. No NIH could be detected in 33% of patients. CONCLUSIONS: EES implantation in CTO patients is associated with high rates of angiographic restenosis as well as revascularization, yet most patients derived significant symptomatic improvement despite focal NIH formation.Item Large Scale Profiling of Fibroblasts from Pediatric Patients with Inborn Errors of Metabolism Results in the Identification of Siblings with L-2-Hydroxyglutaric Aciduria(2020-01-21) Franklin, Jordan; Gu, Wen; Ni, Min; DeBerardinis, RalphInborn errors of metabolism (IEMs) are caused by germline mutations that interfere with the normal physiological functioning of single metabolic enzymes or nutrient transporters1. Discerning an effective treatment for IEMs requires identifying, validating and understanding the impact of disease-causing mutations that are specific to the patient1. Whole-exome sequencing (WES) is the most common strategy for identifying unknown genetic aberrations1. Metabolomics and metabolic flux analysis (MFA) can help functionally validate whether suspected mutations are causative as well as direct the development of diagnostic and therapeutic approaches1. More specifically, mass spectrometry-based metabolomics enables quantification of metabolite abundance, and isotopically labeled MFA enables assessment of the precise manner in which patient cells utilize nutrients for various catabolic pathways1,2. In this study, we conducted metabolomics and MFA to assess metabolic abnormalities in fibroblasts derived from 27 patients, who possessed both known and unknown IEMs, and to gain mechanistic insight into these diseases. To characterize the metabolic abnormalities, we performed a metabolomics experiment that involved 27 patient-derived fibroblast lines and 4 control lines, which eventually yielded a detailed metabolic profile (681 metabolites) for each sample. Additionally, 13C-labeled glucose and 13C-labeled glutamine were introduced to various cell lines in order to evaluate the flux of glucose and glutamine metabolism. We confirmed the quality of our metabolomics results by conducting unsupervised clustering analysis, which showed close grouping of biological replicates, as well as by validating the metabolic changes in a previously reported patient1. Differential analysis of metabolites for each of the 27 patients provided further insight into their specific metabolic anomalies. Notably, we discovered a pair of siblings with dramatically increased 2-hydroxyglutarate (2-HG) levels. By using a derivatization-based mass spectrometry method, we determined that L-2-HG rather than D-2-HG was elevated in the patient samples. WES of both children identified a homozygous mutation that introduced a stop codon in the L-2-HG dehydrogenase gene. This was consistent with the diagnosis of L-2-hydroxyglutaric aciduria in the children3. Metabolic flux analysis provided mechanistic insight into the disease, showing that L-2-HG was primarily being made from glutamine rather than glucose. The unbiased metabolomics approach enabled the discovery and characterization of an important metabolic deficiency and will likely contribute to the discovery of additional IEMs in the cohort.Item Presence of Bland Thrombus Is a Negative Indicator for Cancer Specific Survival in Patients Undergoing Nephrectomy for Kidney Tumors with Venous Tumor Thrombus(2016-01-19) Rew, Charles; Chen, Gong; Hutchinson, Ryan; Singla, Nirmish; Meissner, Matthew; Sheth, Kunj; Haddad, Ahmed; Mann, Michael; Abel, E. Jason; Margulis, Vitaly; Thompson, HoustonPURPOSE: We sought to evaluate the oncologic outcomes of patients undergoing nephrectomy for tumors with venous tumor thrombus with respect to presence or absence of bland thrombus. METHODS: Multi-institutional, IRB approved retrospective nephrectomy databases were reviewed for identification of patients with and without bland thrombus identified on preoperative imaging, intraoperatively, or during final pathologic evaluation. RESULTS: 388 patients were identified including 225 without bland thrombus and 163 with bland thrombus. Median patient age was 62 and median ECOG performance status was 1. Median survival time for tumors without bland thrombus was 76.7 months (95% CI: 57.7 - 95.8) versus 28.3 months (95% CI: 23.0 - 33.5) for those with bland thrombus. Bland thrombus was not associated with histologic subtype (p = 0.069) or sarcomatoid differentiation (p= 0.60) and was highly associated with tumor stage (p<0.001), level of tumor thrombus (p<0.001) and positive margin status (p<0.001). Presence of bland thrombus was associated with decreased cancer-specific survival (HR 2.03, p < 0.001) CONCLUSION: Presence of bland tumor thrombus is associated with adverse pathologic features and inferior oncologic outcomes in patients treated for RCC and venous tumor thrombus. Pre-surgical identification of bland tumor thrombus may play a role in patient counseling and selection for surgery.