69 results on '"Harris PS"'
Search Results
2. Radiation Dose Estimation in the 1958 Los Alamos Criticality Accident
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Harris Ps
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Physics ,Epidemiology ,business.industry ,Health, Toxicology and Mutagenesis ,Lethal dose ,Poison control ,Film badge dosimeter ,Radiation ,Radiation Dosage ,Neutron temperature ,Accidents ,Humans ,Radiology, Nuclear Medicine and imaging ,Neutron ,Irradiation ,Radiation Injuries ,Radiometry ,Nuclear medicine ,business ,Plutonium-239 - Abstract
An accidental radiation excursion occurring durin Pu/sup 2//sup 3//sup 9/ recovery operation resulted in significant exposure of three personnel, one exposure terminating in fatality. Irradiation of the fatally exposed person was nonuniform, and it was possible only to estimate his average body dose and dose incident to specific body areas. His average fast neutron dose was estimated as about 900 rads, and gamma dose as 3000 to 4000 rads. The fast neutron doses incident to the head and upper abdomen were estimated as 2600 and 3000 rads, respectively, and the respective incident gamma doses as about 7800 and 9000 rads. A second operator received a neutron dose estimated at 2.6 rads, and his film badge showed a gamma of 118 rads. The third person's neutron and gamma doses were estimated as 1.4 and 31.5 rads, respectively. Most of the gamma dose to the nonfatality exposed operators was accumulated during subsequent trips near the source. (auth)
- Published
- 1961
3. Quantifying Protein Acetylation in Diabetic Nephropathy from Formalin-Fixed Paraffin-Embedded Tissue.
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Schwab SK, Harris PS, Michel C, McGinnis CD, Nahomi RB, Assiri MA, Reisdorph R, Henriksen K, Orlicky DJ, Levi M, Rosenberg A, Nagaraj RH, and Fritz KS
- Abstract
Purpose: Diabetic kidney disease (DKD) is a serious complication of diabetes mellitus and a leading cause of chronic kidney disease and end-stage renal disease. One potential mechanism underlying cellular dysfunction contributing to kidney disease is aberrant protein post-translational modifications. Lysine acetylation is associated with cellular metabolic flux and is thought to be altered in patients with diabetes and dysfunctional renal metabolism., Experimental Design: A novel extraction and LC-MS/MS approach was adapted to quantify sites of lysine acetylation from formalin-fixed paraffin-embedded (FFPE) kidney tissue and from patients with DKD and non-diabetic donors (n = 5 and n = 7, respectively)., Results: Analysis of FFPE tissues identified 840 total proteins, with 225 of those significantly changing in patients with DKD. Acetylomic analysis quantified 289 acetylated peptides, with 69 of those significantly changing. Pathways impacted in DKD patients revealed numerous metabolic pathways, specifically mitochondrial function, oxidative phosphorylation, and sirtuin signaling. Differential protein acetylation in DKD patients impacted sirtuin signaling, valine, leucine, and isoleucine degradation, lactate metabolism, oxidative phosphorylation, and ketogenesis., Conclusions and Clinical Relevance: A quantitative acetylomics platform was developed for protein biomarker discovery in formalin-fixed and paraffin-embedded biopsies of kidney transplant patients suffering from DKD., (© 2024 Wiley‐VCH GmbH.)
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- 2024
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4. Click chemistry-based thiol redox proteomics reveals significant cysteine reduction induced by chronic ethanol consumption.
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Harris PS, McGinnis CD, Michel CR, Marentette JO, Reisdorph R, Roede JR, and Fritz KS
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- Mice, Animals, Proteome metabolism, Proteomics, Tandem Mass Spectrometry, Click Chemistry, Oxidation-Reduction, Ethanol, Cysteine metabolism, Sulfhydryl Compounds metabolism
- Abstract
In the U.S., alcohol-associated liver disease (ALD) impacts millions of people and is a major healthcare burden. While the pathology of ALD is unmistakable, the molecular mechanisms underlying ethanol hepatotoxicity are not fully understood. Hepatic ethanol metabolism is intimately linked with alterations in extracellular and intracellular metabolic processes, specifically oxidation/reduction reactions. The xenobiotic detoxification of ethanol leads to significant disruptions in glycolysis, β-oxidation, and the TCA cycle, as well as oxidative stress. Perturbation of these regulatory networks impacts the redox status of critical regulatory protein thiols throughout the cell. Integrating these key concepts, our goal was to apply a cutting-edge approach toward understanding mechanisms of ethanol metabolism in disrupting hepatic thiol redox signaling. Utilizing a chronic murine model of ALD, we applied a cysteine targeted click chemistry enrichment coupled with quantitative nano HPLC-MS/MS to assess the thiol redox proteome. Our strategy reveals that ethanol metabolism largely reduces the cysteine proteome, with 593 cysteine residues significantly reduced and 8 significantly oxidized cysteines. Ingenuity Pathway Analysis demonstrates that ethanol metabolism reduces specific cysteines throughout ethanol metabolism (Adh1, Cat, Aldh2), antioxidant pathways (Prx1, Mgst1, Gsr), as well as many other biochemical pathways. Interestingly, a sequence motif analysis of reduced cysteines showed a correlation for hydrophilic, charged amino acids lysine or glutamic acid nearby. Further research is needed to determine how a reduced cysteine proteome impacts individual protein activity across these protein targets and pathways. Additionally, understanding how a complex array of cysteine-targeted post-translational modifications (e.g., S-NO, S-GSH, S-OH) are integrated to regulate redox signaling and control throughout the cell is key to the development of redox-centric therapeutic agents targeted to ameliorate the progression of ALD., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2023
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5. Redox state and the sirtuin deacetylases are major factors that regulate the acetylation status of the stress protein NQO1.
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Siegel D, Harris PS, Michel CR, de Cabo R, Fritz KS, and Ross D
- Abstract
The stress induced protein NQO1 can participate in a wide range of biological pathways which are dependent upon the interaction of NQO1 with protein targets. Many of the protein-protein interactions involving NQO1 have been shown to be regulated by the pyridine nucleotide redox balance. NQO1 can modify its conformation as a result of redox changes in pyridine nucleotides and sites on the C-terminal and helix seven regions of NQO1 have been identified as potential areas that may be involved in redox-dependent protein-protein interactions. Since post-translational modifications can modify the functionality of proteins, we examined whether redox-dependent conformational changes induced in NQO1 would alter lysine acetylation. Recombinant NQO1 was incubated with and without NADH then acetylated non-enzymatically by acetic anhydride or S-acetylglutathione (Ac-GSH). NQO1 acetylation was determined by immunoblot and site-specific lysine acetylation was quantified by mass spectrometry (MS). NQO1 was readily acetylated by acetic anhydride and Ac-GSH. Interestingly, despite a large number of lysine residues (9%) in NQO1 only a small subset of lysines were acetylated and the majority of these were located in or near the functional C-terminal or helix seven regions. Reduction of NQO1 by NADH prior to acetylation resulted in almost complete protection of NQO1 from lysine acetylation as confirmed by immunoblot analysis and MS. Lysines located within the redox-active C-terminus and helix seven regions were readily acetylated when NQO1 was in an oxidized conformation but were protected from acetylation when NQO1 was in the reduced conformation. To investigate regulatory mechanisms of enzymatic deacetylation, NQO1 was acetylated by Ac-GSH then exposed to purified sirtuins (SIRT 1-3) or histone deacetylase 6 (HDAC6). NQO1 could be deacetylated by all sirtuin isoforms and quantitative MS analysis performed using SIRT2 revealed very robust deacetylation of NQO1, specifically at K
262 and K271 in the C-terminal region. No deacetylation of NQO1 by HDAC6 was detected. These data demonstrate that the same subset of key lysine residues in the C-terminal and helix seven regions of NQO1 undergo redox dependent acetylation and are regulated by sirtuin-mediated deacetylation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Siegel, Harris, Michel, de Cabo, Fritz and Ross.)- Published
- 2022
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6. Advancing proteomics and machine learning in the clinic: an editorial on " Noninvasive proteomic biomarkers for alcohol-related liver disease ".
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McGinnis CD, Graham BIM, Harris PS, and Fritz KS
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Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://hbsn.amegroups.com/article/view/10.21037/hbsn-22-390/coif). The authors have no conflicts of interest to declare.
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- 2022
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7. Biochemical Mechanisms of Sirtuin-Directed Protein Acylation in Hepatic Pathologies of Mitochondrial Dysfunction.
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McGinnis CD, Jennings EQ, Harris PS, Galligan JJ, and Fritz KS
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- Acetylation, Mitochondria metabolism, Mitochondrial Proteins metabolism, Sirtuin 3 metabolism, Sirtuins metabolism
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Mitochondrial protein acetylation is associated with a host of diseases including cancer, Alzheimer's, and metabolic syndrome. Deciphering the mechanisms regarding how protein acetylation contributes to disease pathologies remains difficult due to the complex diversity of pathways targeted by lysine acetylation. Specifically, protein acetylation is thought to direct feedback from metabolism, whereby nutritional status influences mitochondrial pathways including beta-oxidation, the citric acid cycle, and the electron transport chain. Acetylation provides a crucial connection between hepatic metabolism and mitochondrial function. Dysregulation of protein acetylation throughout the cell can alter mitochondrial function and is associated with numerous liver diseases, including non-alcoholic and alcoholic fatty liver disease, steatohepatitis, and hepatocellular carcinoma. This review introduces biochemical mechanisms of protein acetylation in the regulation of mitochondrial function and hepatic diseases and offers a viewpoint on the potential for targeted therapies.
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- 2022
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8. Proteomic analysis of alcohol-associated hepatitis reveals glycoprotein NMB (GPNMB) as a novel hepatic and serum biomarker.
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Harris PS, Michel CR, Yun Y, McGinnis CD, Assiri MA, Ahmadi AR, Sun Z, Roede JR, Burchill MA, Orlicky DJ, McCullough RL, and Fritz KS
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- Animals, Biomarkers metabolism, Eye Proteins metabolism, Glycoproteins metabolism, Humans, Liver metabolism, Membrane Glycoproteins metabolism, Mice, Tandem Mass Spectrometry, Hepatitis, Alcoholic, Proteomics
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Alcohol consumption remains a leading cause of liver disease worldwide, resulting in a complex array of hepatic pathologies, including steatosis, steatohepatitis, and cirrhosis. Individuals who progress to a rarer form of alcohol-associated liver disease (ALD), alcohol-associated hepatitis (AH), require immediate life-saving intervention in the form of liver transplantation. Rapid onset of AH is poorly understood and the metabolic mechanisms contributing to the progression to liver failure remain undetermined. While multiple mechanisms have been identified that contribute to ALD, no cures exist and mortality from AH remains high. To identify novel pathways associated with AH, our group utilized proteomics to investigate AH-specific biomarkers in liver explant tissues. The goal of the present study was to determine changes in the proteome as well as epigenetic changes occurring in AH. Protein abundance and acetylomic analyses were performed utilizing nHPLC-MS/MS, revealing significant changes to proteins associated with metabolic and inflammatory fibrosis pathways. Here, we describe a novel hepatic and serum biomarker of AH, glycoprotein NMB (GPNMB). The anti-inflammatory protein GPNMB was significantly increased in AH explant liver and serum compared to healthy donors by 50-fold and 6.5-fold, respectively. Further, bioinformatics analyses identified an AH-dependent decrease in protein abundance across fatty acid degradation, biosynthesis of amino acids, and carbon metabolism. The greatest increases in protein abundance were observed in pathways for focal adhesion, lysosome, phagosome, and actin cytoskeleton. In contrast with the hyperacetylation observed in murine models of ALD, protein acetylation was decreased in AH compared to normal liver across fatty acid degradation, biosynthesis of amino acids, and carbon metabolism. Interestingly, immunoblot analysis found epigenetic marks were significantly increased in AH explants, including Histone H3K9 and H2BK5 acetylation. The increased acetylation of histones likely plays a role in the altered proteomic profile observed, including increases in GPNMB. Indeed, our results reveal that the AH proteome is dramatically impacted through unanticipated and unknown mechanisms. Understanding the origin and consequences of these changes will yield new mechanistic insight for ALD as well as identify novel hepatic and serum biomarkers, such as GPNMB., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2021 Elsevier Inc. All rights reserved.)
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- 2022
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9. Cystathionine γ-lyase promotes estrogen-stimulated uterine artery blood flow via glutathione homeostasis.
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Bok R, Guerra DD, Lorca RA, Wennersten SA, Harris PS, Rauniyar AK, Stabler SP, MacLean KN, Roede JR, Brown LD, and Hurt KJ
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- Animals, Estrogens, Female, Glutathione, Homeostasis, Mice, Pregnancy, Uterine Artery, Cystathionine gamma-Lyase genetics, Hydrogen Sulfide
- Abstract
During pregnancy, estrogen (E
2 ) stimulates uterine artery blood flow (UBF) by enhancing nitric oxide (NO)-dependent vasodilation. Cystathionine γ-lyase (CSE) promotes vascular NO signaling by producing hydrogen sulfide (H2 S) and by maintaining the ratio of reduced-to-oxidized intracellular glutathione (GSH/GSSG) through l-cysteine production. Because redox homeostasis can influence NO signaling, we hypothesized that CSE mediates E2 stimulation of UBF by modulating local intracellular cysteine metabolism and GSH/GSSG levels to promote redox homeostasis. Using non-pregnant ovariectomized WT and CSE-null (CSE KO) mice, we performed micro-ultrasound of mouse uterine and renal arteries to assess changes in blood flow upon exogenous E2 stimulation. We quantified serum and uterine artery NO metabolites (NOx ), serum amino acids, and uterine and renal artery GSH/GSSG. WT and CSE KO mice exhibited similar baseline uterine and renal blood flow. Unlike WT, CSE KO mice did not exhibit expected E2 stimulation of UBF. Renal blood flow was E2 -insensitive for both genotypes. While serum and uterine artery NOx were similar between genotypes at baseline, E2 decreased NOx in CSE KO serum. Cysteine was also lower in CSE KO serum, while citrulline and homocysteine levels were elevated. E2 and CSE deletion additively decreased GSH/GSSG in uterine arteries. In contrast, renal artery GSH/GSSG was insensitive to E2 or CSE deletion. Together, these findings suggest that CSE maintenance of uterine artery GSH/GSSG facilitates nitrergic signaling in uterine arteries and is required for normal E2 stimulation of UBF. These data have implications for pregnancy pathophysiology and the selective hormone responses of specific vascular beds., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)- Published
- 2021
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10. Intervention development for exercise promotion at active charity events in the UK.
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Jones BA, Munir F, Harris PS, Bhatnagar P, and Stevinson C
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- Exercise, Humans, Needs Assessment, United Kingdom, Charities, Health Promotion
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This study used the Intervention Mapping protocol to design an evidence-based intervention package for organizers of active charity events to support their participants in remaining or becoming regular exercisers. A mixed-methods approach following the Intervention Mapping protocol was used to develop intervention components. A needs assessment was initially performed to identify the behavioural and environmental determinants of exercise for charity event participants (Step 1). Next, the intended intervention outcomes, and performance and change objectives were specified (Step 2). Theory-based change methods were selected and matched with practical strategies (Step 3). This resulted in the design of the first iteration of the intervention which underwent pre-testing with former event participants and feasibility testing at an active charity event (Step 4). The evidence-based interventions included components to implement at events (e.g. an activity and information zone, and exercise planner), along with elements pre- and post-event (e.g. social media). Pre-testing indicated high acceptability of the planned components, but feasibility testing suggested low engagement with the intervention. Despite developing the intervention package through the systematic process of Intervention Mapping, preliminary data suggest that further development and testing is needed to refine the intervention before implementation., (© The Author(s) 2020. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.)
- Published
- 2020
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11. Gastrointestinal Stromal Tumor: GIST Another Duodenal Ulcer.
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Harris PS, Romano J, Russ KB, Shoreibah MG, and Baig KRKK
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Background: Gastrointestinal stromal tumors (GISTs), although exceedingly rare, are the most common mesenchymal tumors in the gastrointestinal (GI) tract. GISTs are often asymptomatic; approximately 10% are found incidentally on imaging or endoscopy for other indications, although GI bleeding, intestinal obstruction, and perforation can occur. We present a case of upper GI bleeding from a duodenal GIST. Proton-pump inhibitor (PPI) therapy resulted in complete endoscopic ulcer healing, yet a discrete mass lesion was identified on endoscopic ultrasound (EUS). Case Report: A 70-year-old female presented with upper GI bleeding, and a duodenal ulcer was identified with esophagogastroduodenoscopy (EGD). Computed tomography (CT) scan of the abdomen and pelvis showed duodenal bulb thickening without clear mass. The ulcer was treated with 1:10,000 concentration epinephrine, injected in 4 quadrants around the ulcer base. The patient's GI bleeding resolved, and she was discharged with a referral for outpatient EUS follow-up. One month later, EUS showed resolution of the ulcer after PPI therapy but also showed a lesion consistent with GIST that was confirmed by cytologic analysis. The patient was started on imatinib therapy and had no further bleeding. Conclusion: Initial EGD and CT findings could have easily been attributed to duodenal peptic ulcer disease for which follow-up endoscopy is not routinely recommended given the low risk of malignancy. However, because of the high index of suspicion on the part of the referring physicians, duodenal GIST was diagnosed. This case extends the spectrum of the presentation, evaluation, and diagnosis of GISTs and stresses the importance of keeping this rare disease on the provider's differential, even after routine workup shows no findings of tumor., (©2020 by the author(s); Creative Commons Attribution License (CC BY).)
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- 2020
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12. Investigating RNA expression profiles altered by nicotinamide mononucleotide therapy in a chronic model of alcoholic liver disease.
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Assiri MA, Ali HR, Marentette JO, Yun Y, Liu J, Hirschey MD, Saba LM, Harris PS, and Fritz KS
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- Alanine Transaminase blood, Animals, Aspartate Aminotransferases blood, Chronic Disease, Disease Models, Animal, Ethanol, Gene Expression Regulation drug effects, Liver Diseases, Alcoholic blood, Liver Diseases, Alcoholic genetics, Metabolome, Metabolomics, Mice, Inbred C57BL, Nicotinamide Mononucleotide pharmacology, Protective Agents metabolism, RNA metabolism, Signal Transduction drug effects, Gene Expression Profiling, Liver Diseases, Alcoholic drug therapy, Nicotinamide Mononucleotide therapeutic use, RNA genetics
- Abstract
Background: Chronic alcohol consumption is a significant cause of liver disease worldwide. Several biochemical mechanisms have been linked to the initiation and progression of alcoholic liver disease (ALD) such as oxidative stress, inflammation, and metabolic dysregulation, including the disruption of NAD
+ /NADH. Indeed, an ethanol-mediated reduction in hepatic NAD+ levels is thought to be one factor underlying ethanol-induced steatosis, oxidative stress, steatohepatitis, insulin resistance, and inhibition of gluconeogenesis. Therefore, we applied a NAD+ boosting supplement to investigate alterations in the pathogenesis of early-stage ALD., Methods: To examine the impact of NAD+ therapy on the early stages of ALD, we utilized nicotinamide mononucleotide (NMN) at 500 mg/kg intraperitoneal injection every other day, for the duration of a Lieber-DeCarli 6-week chronic ethanol model in mice. Numerous strategies were employed to characterize the effect of NMN therapy, including the integration of RNA-seq, immunoblotting, and metabolomics analysis., Results: Our findings reveal that NMN therapy increased hepatic NAD+ levels, prevented an ethanol-induced increase in plasma ALT and AST, and changed the expression of 25% of the genes that were modulated by ethanol metabolism. These genes were associated with a number of pathways including the MAPK pathway. Interestingly, our analysis revealed that NMN treatment normalized Erk1/2 signaling and prevented an induction of Atf3 overexpression., Conclusions: These findings reveal previously unreported mechanisms by which NMN supplementation alters hepatic gene expression and protein pathways to impact ethanol hepatotoxicity in an early-stage murine model of ALD. Overall, our data suggest further research is needed to fully characterize treatment paradigms and biochemical implications of NAD+ -based interventions.- Published
- 2019
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13. SNARE proteins rescue impaired autophagic flux in Down syndrome.
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Aivazidis S, Jain A, Rauniyar AK, Anderson CC, Marentette JO, Orlicky DJ, Fritz KS, Harris PS, Siegel D, Maclean KN, and Roede JR
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- Cell Line, Female, Gene Expression Regulation, Humans, Infant, Infant, Newborn, Lysosomal-Associated Membrane Protein 2 metabolism, Lysosomes metabolism, Male, Microtubule-Associated Proteins metabolism, Protein Transport, R-SNARE Proteins metabolism, RNA-Binding Proteins metabolism, Autophagy, Down Syndrome metabolism, Down Syndrome pathology, SNARE Proteins metabolism
- Abstract
Down syndrome (DS) is a chromosomal disorder caused by trisomy of chromosome 21 (Ts21). Unbalanced karyotypes can lead to dysfunction of the proteostasis network (PN) and disrupted proteostasis is mechanistically associated with multiple DS comorbidities. Autophagy is a critical component of the PN that has not previously been investigated in DS. Based on our previous observations of PN disruption in DS, we investigated possible dysfunction of the autophagic machinery in human DS fibroblasts and other DS cell models. Following induction of autophagy by serum starvation, DS fibroblasts displayed impaired autophagic flux indicated by autophagolysosome accumulation and elevated p62, NBR1, and LC3-II abundance, compared to age- and sex-matched, euploid (CTL) fibroblasts. While lysosomal physiology was unaffected in both groups after serum starvation, we observed decreased basal abundance of the Soluble N-ethylmaleimide-sensitive-factor Attachment protein Receptor (SNARE) family members syntaxin 17 (STX17) and Vesicle Associated Membrane Protein 8 (VAMP8) indicating that decreased autophagic flux in DS is due at least in part to a possible impairment of autophagosome-lysosome fusion. This conclusion was further supported by the observation that over-expression of either STX17 or VAMP8 in DS fibroblasts restored autophagic degradation and reversed p62 accumulation. Collectively, our results indicate that impaired autophagic clearance is a characteristic of DS cells that can be reversed by enhancement of SNARE protein expression and provides further evidence that PN disruption represents a candidate mechanism for multiple aspects of pathogenesis in DS and a possible future target for therapeutic intervention., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2019
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14. Cholestatic liver disease results increased production of reactive aldehydes and an atypical periportal hepatic antioxidant response.
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Shearn CT, Fennimore B, Orlicky DJ, Gao YR, Saba LM, Battista KD, Aivazidis S, Assiri M, Harris PS, Michel C, Merrill GF, Schmidt EE, Colgan SP, and Petersen DR
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- Adult, Animals, Antioxidants metabolism, Female, Glutathione Transferase metabolism, Humans, Inflammation, Liver physiopathology, Lysosomes metabolism, Mice, Mice, Inbred C57BL, Mice, Knockout, Middle Aged, NF-kappa B metabolism, Oxidation-Reduction, Proteomics, Ribosomes metabolism, Signal Transduction, Superoxide Dismutase metabolism, Up-Regulation, Aldehydes metabolism, Antioxidants pharmacology, Cholestasis metabolism, Liver metabolism, Liver Diseases metabolism, Oxidative Stress
- Abstract
Cholangiopathies such as primary sclerosing cholangitis (PSC) are chronic liver diseases characterized by increased cholestasis, biliary inflammation and oxidative stress. The objective of this study was to elucidate the impact of cholestatic injury on oxidative stress-related factors. Using hepatic tissue and whole cell liver extracts (LE) isolated from 11-week old C57BL/6J (WT) and Mdr2
KO mice, inflammation and oxidative stress was assessed. Concurrently, specific targets of carbonylation were assessed in LE prepared from murine groups as well as from normal and human patients with end-stage PSC. Identified carbonylated proteins were further evaluated using bioinformatics analyses. Picrosirius red staining revealed extensive fibrosis in Mdr2KO liver, and fibrosis colocalized with increased periportal inflammatory cells and both acrolein and 4-HNE staining. Western blot analysis revealed elevated periportal expression of antioxidant proteins Cbr3, GSTμ, Prdx5, TrxR1 and HO-1 but not GCLC, GSTπ or catalase in the Mdr2KO group when compared to WT. From immunohistochemical analysis, increased periportal reactive aldehyde production colocalized with elevated staining of Cbr3, GSTμ and TrxR1 but surprisingly not with Nrf2. Mass spectrometric analysis revealed an increase in carbonylated proteins in the Mdr2KO and PSC groups compared to respective controls. Gene ontology and KEGG pathway analysis of carbonylated proteins revealed a propensity for increased carbonylation of proteins broadly involved in metabolic processes as well more specifically in Rab-mediated signal transduction, lysosomes and the large ribosomal subunit in human PSC. Western blot analysis of Rab-GTPase expression revealed no significant differences in Mdr2KO mice when compared to WT livers. In contrast, PSC tissue exhibited decreased levels of Rabs 4, 5 and increased abundance of Rabs 6 and 9a protein. Results herein reveal that cholestasis induces stage-dependent increases in periportal oxidative stress responses and protein carbonylation, potentially contributing to pathogenesis in Mdr2KO . Furthermore, during early stage cholestasis, there is cell-specific upregulation of some but not all, antioxidant proteins., (Copyright © 2019 Elsevier Inc. All rights reserved.)- Published
- 2019
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15. Barriers and facilitators to screening and treating malnutrition in older adults living in the community: a mixed-methods synthesis.
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Harris PS, Payne L, Morrison L, Green SM, Ghio D, Hallett C, Parsons EL, Aveyard P, Roberts HC, Sutcliffe M, Robinson S, Slodkowska-Barabasz J, Little PS, Stroud MA, and Yardley L
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- Aged, Humans, Independent Living, Malnutrition diagnosis, Malnutrition diet therapy, Mass Screening methods, Primary Health Care
- Abstract
Background: Malnutrition (specifically undernutrition) in older, community-dwelling adults reduces well-being and predisposes to disease. Implementation of screen-and-treat policies could help to systematically detect and treat at-risk and malnourished patients. We aimed to identify barriers and facilitators to implementing malnutrition screen and treat policies in primary/community care, which barriers have been addressed and which facilitators have been successfully incorporated in existing interventions., Method: A data-base search was conducted using MEDLINE, Embase, PsycINFO, DARE, CINAHL, Cochrane Central and Cochrane Database of Systematic Reviews from 2012 to June 2016 to identify relevant qualitative and quantitative literature from primary/community care. Studies were included if participants were older, community-dwelling adults (65+) or healthcare professionals who would screen and treat such patients. Barriers and facilitators were extracted and mapped onto intervention features to determine whether these had addressed barriers., Results: Of a total of 2182 studies identified, 21 were included (6 qualitative, 12 quantitative and 3 mixed; 14 studies targeting patients and 7 targeting healthcare professionals). Facilitators addressing a wide range of barriers were identified, yet few interventions addressed psychosocial barriers to screen-and-treat policies for patients, such as loneliness and reluctance to be screened, or healthcare professionals' reservations about prescribing oral nutritional supplements., Conclusion: The studies reviewed identified several barriers and facilitators and addressed some of these in intervention design, although a prominent gap appeared to be psychosocial barriers. No single included study addressed all barriers or made use of all facilitators, although this appears to be possible. Interventions aiming to implement screen-and-treat approaches to malnutrition in primary care should consider barriers that both patients and healthcare professionals may face., Review Registrations: PROSPERO: CRD42017071398 . The review protocol was registered retrospectively.
- Published
- 2019
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16. The absence of SIRT3 and SIRT5 promotes the acetylation of lens proteins and improves the chaperone activity of α-crystallin in mouse lenses.
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Nandi SK, Nahomi RB, Harris PS, Michel CR, Fritz KS, and Nagaraj RH
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- Acetylation, Animals, Blotting, Western, Cataract metabolism, Disease Models, Animal, Mass Spectrometry, Mice, Mice, Inbred C57BL, Mice, Knockout, RNA genetics, Sirtuin 3 biosynthesis, Sirtuins biosynthesis, alpha-Crystallins metabolism, Cataract genetics, Gene Expression Regulation, Lens, Crystalline metabolism, Molecular Chaperones metabolism, Sirtuin 3 genetics, Sirtuins genetics, alpha-Crystallins genetics
- Abstract
Acetylation of lysine residues occurs in lens proteins. Previous studies have shown an improvement in the chaperone activity of αA-crystallin upon acetylation. Sirtuins are NAD
+ -dependent enzymes that can deacylate proteins. The roles of sirtuins in regulating the acetylation of lens proteins and their impacts on the function of α-crystallin are not known. Here, we detected sirtuin activity in mouse lenses, and SIRT3 and SIRT5 were present primarily in the mitochondria of cultured primary mouse lens epithelial cells. Western blotting showed higher levels of protein acetylation in the lenses of SIRT3 KO and SIRT5 KO mice than in lenses of WT mice. Mass spectrometry analyses revealed a greater number of acetylated lysine residues in α-crystallin isolated from the SIRT3 and SIRT5 KO lenses than from WT lenses. α-Crystallin isolated from SIRT3 and SIRT5 KO lenses displayed a higher surface hydrophobicity and higher chaperone activity than the protein isolated from WT lenses. Thus, SIRTs regulate the acetylation levels of crystallins in mouse lenses, and acetylation in lenses enhances the chaperone activity of α-crystallin., (Copyright © 2019 Elsevier Ltd. All rights reserved.)- Published
- 2019
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17. Hepatocellular carcinoma surveillance: An evidence-based approach.
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Harris PS, Hansen RM, Gray ME, Massoud OI, McGuire BM, and Shoreibah MG
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- Biomarkers, Tumor blood, Carcinoma, Hepatocellular blood, Carcinoma, Hepatocellular pathology, Carcinoma, Hepatocellular virology, Disease Progression, Early Detection of Cancer standards, Evidence-Based Medicine standards, Guideline Adherence, Hepatitis B virology, Humans, Liver diagnostic imaging, Liver pathology, Liver Cirrhosis virology, Liver Neoplasms blood, Liver Neoplasms pathology, Liver Neoplasms virology, Magnetic Resonance Imaging, Practice Guidelines as Topic, Risk Factors, Sensitivity and Specificity, Socioeconomic Factors, Tomography, X-Ray Computed, Ultrasonography, Carcinoma, Hepatocellular diagnosis, Early Detection of Cancer methods, Evidence-Based Medicine methods, Hepatitis B pathology, Liver Cirrhosis pathology, Liver Neoplasms diagnosis
- Abstract
Hepatocellular carcinoma (HCC) makes up 75%-85% of all primary liver cancers and is the fourth most common cause of cancer related death worldwide. Chronic liver disease is the most significant risk factor for HCC with 80%-90% of new cases occurring in the background of cirrhosis. Studies have shown that early diagnosis of HCC through surveillance programs improve prognosis and availability of curative therapies. All patients with cirrhosis and high-risk hepatitis B patients are at risk for HCC and should undergo surveillance. The recommended surveillance modality is abdominal ultrasound (US) given that it is cost effective and noninvasive with good sensitivity. However, US is limited in obese patients and those with non-alcoholic fatty liver disease (NAFLD). With the current obesity epidemic and rise in the prevalence of NAFLD, abdominal computed tomography or magnetic resonance imaging may be indicated as the primary screening modality in these patients. The addition of alpha-fetoprotein to a surveillance regimen is thought to improve the sensitivity of HCC detection. Further investigation of serum biomarkers is needed. Semiannual screening is the suggested surveillance interval. Surveillance for HCC is underutilized and low adherence disproportionately affects certain demographics such as non-Caucasian race and low socioeconomic status., Competing Interests: Conflict-of-interest statement: No potential conflicts of interest.
- Published
- 2019
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18. Quantifying Competition among Mitochondrial Protein Acylation Events Induced by Ethanol Metabolism.
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Ali HR, Assiri MA, Harris PS, Michel CR, Yun Y, Marentette JO, Huynh FK, Orlicky DJ, Shearn CT, Saba LM, Reisdorph R, Reisdorph N, Hirschey MD, and Fritz KS
- Subjects
- Animals, Liver Diseases, Alcoholic metabolism, Male, Metabolic Networks and Pathways drug effects, Mice, Mice, Knockout, Sirtuin 3 genetics, Sirtuin 3 metabolism, Sirtuins genetics, Sirtuins metabolism, Acylation drug effects, Ethanol pharmacology, Mitochondria drug effects, Mitochondria metabolism, Proteome chemistry, Proteome drug effects, Proteome metabolism
- Abstract
Mitochondrial dysfunction is one of many key factors in the etiology of alcoholic liver disease (ALD). Lysine acetylation is known to regulate numerous mitochondrial metabolic pathways, and recent reports demonstrate that alcohol-induced protein acylation negatively impacts these processes. To identify regulatory mechanisms attributed to alcohol-induced protein post-translational modifications, we employed a model of alcohol consumption within the context of wild type (WT), sirtuin 3 knockout (SIRT3 KO), and sirtuin 5 knockout (SIRT5 KO) mice to manipulate hepatic mitochondrial protein acylation. Mitochondrial fractions were examined by label-free quantitative HPLC-MS/MS to reveal competition between lysine acetylation and succinylation. A class of proteins defined as "differential acyl switching proteins" demonstrate select sensitivity to alcohol-induced protein acylation. A number of these proteins reveal saturated lysine-site occupancy, suggesting a significant level of differential stoichiometry in the setting of ethanol consumption. We hypothesize that ethanol downregulates numerous mitochondrial metabolic pathways through differential acyl switching proteins. Data are available via ProteomeXchange with identifier PXD012089.
- Published
- 2019
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19. Taurine treatment prevents derangement of the hepatic γ-glutamyl cycle and methylglyoxal metabolism in a mouse model of classical homocystinuria: regulatory crosstalk between thiol and sulfinic acid metabolism.
- Author
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Maclean KN, Jiang H, Aivazidis S, Kim E, Shearn CT, Harris PS, Petersen DR, Allen RH, Stabler SP, and Roede JR
- Subjects
- Amino Acids metabolism, Animals, Cystathionine beta-Synthase metabolism, Disease Models, Animal, Female, Homocystinuria drug therapy, Homocystinuria pathology, Liver drug effects, Male, Metabolome, Mice, Mice, Inbred C57BL, Oxidation-Reduction, gamma-Glutamyltransferase metabolism, Aminobutyrates metabolism, Homocystinuria metabolism, Liver metabolism, Pyruvaldehyde metabolism, Sulfhydryl Compounds metabolism, Sulfinic Acids metabolism, Taurine pharmacology
- Abstract
Cystathionine β-synthase-deficient homocystinuria (HCU) is a poorly understood, life-threatening inborn error of sulfur metabolism. Analysis of hepatic glutathione (GSH) metabolism in a mouse model of HCU demonstrated significant depletion of cysteine, GSH, and GSH disulfide independent of the block in trans-sulfuration compared with wild-type controls. HCU induced the expression of the catalytic and regulatory subunits of γ-glutamyl ligase, GSH synthase (GS), γ-glutamyl transpeptidase 1, 5-oxoprolinase (OPLAH), and the GSH-dependent methylglyoxal detoxification enzyme, glyoxalase-1. Multiple components of the transcription factor nuclear factor (erythroid-derived 2)-like 2 (Nrf2)-mediated antioxidant-response regulatory axis were induced without any detectable activation of Nrf2. Metabolomic analysis revealed the accumulation of multiple γ-glutamyl amino acids and that plasma ophthalmate levels could serve as a noninvasive marker for hepatic redox stress. Neither cysteine, nor betaine treatment was able to reverse the observed enzyme inductions. Taurine treatment normalized the expression levels of γ-glutamyl ligase C/M, GS, OPLAH, and glyoxalase-1, and reversed HCU-induced deficits in protein glutathionylation by acting to double GSH levels relative to controls. Collectively, our data indicate that the perturbation of the γ-glutamyl cycle could contribute to multiple sequelae in HCU and that taurine has significant therapeutic potential for both HCU and other diseases for which GSH depletion is a critical pathogenic factor.-Maclean, K. N., Jiang, H., Aivazidis, S., Kim, E., Shearn, C. T., Harris, P. S., Petersen, D. R., Allen, R. H., Stabler, S. P., Roede, J. R. Taurine treatment prevents derangement of the hepatic γ-glutamyl cycle and methylglyoxal metabolism in a mouse model of classical homocystinuria: regulatory crosstalk between thiol and sulfinic acid metabolism.
- Published
- 2018
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20. Chronic Ethanol Metabolism Inhibits Hepatic Mitochondrial Superoxide Dismutase via Lysine Acetylation.
- Author
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Assiri MA, Roy SR, Harris PS, Ali H, Liang Y, Shearn CT, Orlicky DJ, Roede JR, Hirschey MD, Backos DS, and Fritz KS
- Subjects
- Acetylation drug effects, Animals, Ethanol administration & dosage, Liver drug effects, Male, Mice, Mice, Inbred C57BL, Mitochondria drug effects, Oxidative Stress drug effects, Oxidative Stress physiology, Superoxide Dismutase antagonists & inhibitors, Ethanol metabolism, Ethanol toxicity, Liver metabolism, Lysine metabolism, Mitochondria metabolism, Superoxide Dismutase metabolism
- Abstract
Background: Chronic ethanol (EtOH) consumption is a major cause of liver disease worldwide. Oxidative stress is a known consequence of EtOH metabolism and is thought to contribute significantly to alcoholic liver disease (ALD). Therefore, elucidating pathways leading to sustained oxidative stress and downstream redox imbalances may reveal how EtOH consumption leads to ALD. Recent studies suggest that EtOH metabolism impacts mitochondrial antioxidant processes through a number of proteomic alterations, including hyperacetylation of key antioxidant proteins., Methods: To elucidate mechanisms of EtOH-induced hepatic oxidative stress, we investigate a role for protein hyperacetylation in modulating mitochondrial superoxide dismutase (SOD2) structure and function in a 6-week Lieber-DeCarli murine model of EtOH consumption. Our experimental approach includes immunoblotting immunohistochemistry (IHC), activity assays, mass spectrometry, and in silico modeling., Results: We found that EtOH metabolism significantly increased the acetylation of SOD2 at 2 functionally relevant lysine sites, K68 and K122, resulting in a 40% decrease in enzyme activity while overall SOD2 abundance was unchanged. In vitro studies also reveal which lysine residues are more susceptible to acetylation. IHC analysis demonstrates that SOD2 hyperacetylation occurs near zone 3 within the liver, which is the main EtOH-metabolizing region of the liver., Conclusions: Overall, the findings presented in this study support a role for EtOH-induced lysine acetylation as an adverse posttranslational modification within the mitochondria that directly impacts SOD2 charge state and activity. Last, the data presented here indicate that protein hyperacetylation may be a major factor contributing to an imbalance in hepatic redox homeostasis due to chronic EtOH metabolism., (Copyright © 2017 by the Research Society on Alcoholism.)
- Published
- 2017
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21. Characterizing Sirtuin 3 Deacetylase Affinity for Aldehyde Dehydrogenase 2.
- Author
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Harris PS, Gomez JD, Backos DS, and Fritz KS
- Subjects
- Acetylation, Aldehyde Dehydrogenase chemistry, Animals, Humans, Lysine chemistry, Mass Spectrometry, Sirtuin 3 chemistry, Aldehyde Dehydrogenase metabolism, Sirtuin 3 metabolism
- Abstract
Mitochondrial aldehyde dehydrogenase (ALDH2) plays a central role in the detoxification of reactive aldehydes generated through endogenous and exogenous sources. The biochemical regulation of enzyme activity through post-translational modification provides an intricate response system regulating mitochondrial detoxification pathways. ALDH2 is a known target of lysine acetylation, which arises as a consequence of mitochondrial bioenergetic flux and sirtuin deacetylase activity. The mitochondrial deacetylase Sirtuin 3 (SIRT3) has been reported to alter ALDH2 lysine acetylation status, yet the mechanism and consequence of this interaction remain unknown. The in vitro results presented here provide a novel biochemical approach using stable-isotope dilution mass spectrometry to elucidate which lysine residues are targeted by SIRT3 for deacetylation. Furthermore, HPLC-MS/MS and computational modeling elucidate a potential role for acetyl-Lys369 on ALDH2 in perturbing normal β-nicotinamide adenine dinucleotide (NAD
+ ) cofactor binding.- Published
- 2017
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22. Increasing inpatient hospice use versus patient preferences in the USA: are patients able to die in the setting of their choice?
- Author
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Hurley SL, Colling C, Bender L, Harris PS, Harrold JK, Teno JM, Ache KA, and Casarett D
- Subjects
- Aged, Aged, 80 and over, Humans, Retrospective Studies, United States, Hospice Care statistics & numerical data, Hospices statistics & numerical data, Inpatients statistics & numerical data, Patient Preference statistics & numerical data
- Abstract
Background: Growth in hospice utilisation has been accompanied by an increase in the proportion of hospice patients who die in an inpatient hospice setting rather than at home., Objective: To determine whether this increase in inpatient utilisation is consistent with patient preferences., Design: Retrospective cohort study., Setting: Seven hospices in the Coalition of Hospices Organised to Investigate Comparative Effectiveness (CHOICE) network., Patients: 70 488 patients admitted between 1 July 2008 and 31 May 2012., Measurements: We measured changes in patients' stated preferences at the time of admission regarding site of death, including weights to adjust for non-response bias. We also assessed patients' actual site of death and concordance with patients' preferences., Results: More patients died receiving inpatient care in 2012 as compared to 2008 (1920 (32.7%), 2537 (18.5%); OR 1.21; 95% CI 1.19 to 1.22; p<0.001). However, patients also expressed an increasing preference for dying in inpatient settings (weighted preferences 27.5% in 2012 vs 7.9% in 2008; p<0.001). The overall proportion of patients who died in the setting of their choice (weighted preferences) increased from 74% in 2008 to 78% in 2012 (p<0.001)., Limitations: This study included only seven hospices, and results may not be representative of the larger hospice population., Conclusions: Although more patients are dying while receiving inpatient care, these changes in site of death seem to reflect changing patient preferences. The net effect is that patients in this sample were more likely to die in the setting of their choice in 2012 than they were in 2008., (Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.)
- Published
- 2017
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23. Quantitative Analysis and Discovery of Lysine and Arginine Modifications.
- Author
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Galligan JJ, Kingsley PJ, Wauchope OR, Mitchener MM, Camarillo JM, Wepy JA, Harris PS, Fritz KS, and Marnett LJ
- Subjects
- HEK293 Cells, Humans, Hydrolysis, Jumonji Domain-Containing Histone Demethylases chemistry, Recombinant Proteins chemistry, Arginine analysis, Chromatography, High Pressure Liquid methods, Histones chemistry, Lysine analysis, Peptides chemistry, Protein Processing, Post-Translational, Tandem Mass Spectrometry methods
- Abstract
Post-translational modifications (PTMs) affect protein function, localization, and stability, yet very little is known about the ratios of these modifications. Here, we describe a novel method to quantitate and assess the relative stoichiometry of Lys and Arg modifications (QuARKMod) in complex biological settings. We demonstrate the versatility of this platform in monitoring recombinant protein modification of peptide substrates, PTMs of individual histones, and the relative abundance of these PTMs as a function of subcellular location. Lastly, we describe a product ion scanning technique that offers the potential to discover unexpected and possibly novel Lys and Arg modifications. In summary, this approach yields accurate quantitation and discovery of protein PTMs in complex biological systems without the requirement of high mass accuracy instrumentation., Competing Interests: The authors declare no competing financial interests.
- Published
- 2017
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24. Checkpoint kinase 1 expression is an adverse prognostic marker and therapeutic target in MYC-driven medulloblastoma.
- Author
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Prince EW, Balakrishnan I, Shah M, Mulcahy Levy JM, Griesinger AM, Alimova I, Harris PS, Birks DK, Donson AM, Davidson N, Remke M, Taylor MD, Handler MH, Foreman NK, Venkataraman S, and Vibhakar R
- Subjects
- Antineoplastic Combined Chemotherapy Protocols pharmacology, Apoptosis drug effects, Benzodiazepinones pharmacology, Biomarkers, Tumor metabolism, Cell Survival drug effects, Cerebellar Neoplasms enzymology, Cerebellar Neoplasms mortality, Cisplatin pharmacology, Disease-Free Survival, Genes, myc, Humans, Kaplan-Meier Estimate, Medulloblastoma enzymology, Medulloblastoma mortality, Prognosis, Pyrazoles pharmacology, Thiophenes pharmacology, Urea analogs & derivatives, Urea pharmacology, Biomarkers, Tumor analysis, Cerebellar Neoplasms pathology, Checkpoint Kinase 1 biosynthesis, Medulloblastoma pathology
- Abstract
Checkpoint kinase 1 (CHK1) is an integral component of the cell cycle as well as the DNA Damage Response (DDR) pathway. Previous work has demonstrated the effectiveness of inhibiting CHK1 with small-molecule inhibitors, but the role of CHK1 mediated DDR in medulloblastoma is unknown. CHK1, both at the mRNA and protein level, is highly expressed in medulloblastoma and elevated CHK1 expression in Group3 medulloblastoma is an adverse prognostic marker. CHK1 inhibition with the small-molecule drug AZD7762, results in decreased cell growth, increased DNA damage and cell apoptosis. Furthermore, AZD7762 acts in synergy with cisplatin in reducing cell proliferation in medulloblastoma. Similar phenotypic changes were observed with another CHK1 inhibitor, PF477736, as well as genetic knockdown using siRNA against CHK1. Treatments with small-molecule inhibitors of CHK1 profoundly modulated the expression of both upstream and downstream target proteins within the CHK1 signaling pathways. This suggests the presence of a feedback loop in activating CHK1. Overall, our results demonstrate that small-molecule inhibition of CHK1 in combination with, cisplatin, is more advantageous than either treatment alone, especially for Group 3 medulloblastoma, and therefore this combined therapeutic approach serves as an avenue for further investigation., Competing Interests: The authors declare no conflicts of interest.
- Published
- 2016
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25. A WEE1 Inhibitor Analog of AZD1775 Maintains Synergy with Cisplatin and Demonstrates Reduced Single-Agent Cytotoxicity in Medulloblastoma Cells.
- Author
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Matheson CJ, Venkataraman S, Amani V, Harris PS, Backos DS, Donson AM, Wempe MF, Foreman NK, Vibhakar R, and Reigan P
- Published
- 2016
- Full Text
- View/download PDF
26. Chronic ethanol consumption induces mitochondrial protein acetylation and oxidative stress in the kidney.
- Author
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Harris PS, Roy SR, Coughlan C, Orlicky DJ, Liang Y, Shearn CT, Roede JR, and Fritz KS
- Subjects
- Acetylation, Alcoholism etiology, Alcoholism pathology, Aldehydes metabolism, Amino Acid Sequence, Amino Acids metabolism, Animals, Glutathione metabolism, Glutathione Reductase metabolism, Glutathione Transferase metabolism, Kidney metabolism, Kidney pathology, Lipid Metabolism drug effects, Male, Mice, Mice, Inbred C57BL, Mitochondria metabolism, Mitochondria pathology, Molecular Sequence Data, Oxidative Phosphorylation drug effects, Peroxiredoxins metabolism, Superoxide Dismutase metabolism, Alcoholism metabolism, Ethanol adverse effects, Kidney drug effects, Mitochondria drug effects, Mitochondrial Proteins metabolism, Oxidative Stress drug effects
- Abstract
In this study, we present the novel findings that chronic ethanol consumption induces mitochondrial protein hyperacetylation in the kidney and correlates with significantly increased renal oxidative stress. A major proteomic footprint of alcoholic liver disease (ALD) is an increase in hepatic mitochondrial protein acetylation. Protein hyperacetylation has been shown to alter enzymatic function of numerous proteins and plays a role in regulating metabolic processes. Renal mitochondrial targets of hyperacetylation include numerous metabolic and antioxidant pathways, such as lipid metabolism, oxidative phosphorylation, and amino acid metabolism, as well as glutathione and thioredoxin pathways. Disruption of protein lysine acetylation has the potential to impair renal function through metabolic dysregulation and decreased antioxidant capacity. Due to a significant elevation in ethanol-mediated renal oxidative stress, we highlight the acetylation of superoxide dismutase, peroxiredoxins, glutathione reductase, and glutathione transferase enzymes. Since oxidative stress is a known factor in ethanol-induced nephrotoxicity, we examined biochemical markers of protein hyperacetylation and oxidative stress. Our results demonstrate increased protein acetylation concurrent with depleted glutathione, altered Cys redox potential, and the presence of 4-HNE protein modifications in our 6-week model of early-stage alcoholic nephrotoxicity. These findings support the hypothesis that ethanol metabolism causes an influx of mitochondrial metabolic substrate, resulting in mitochondrial protein hyperacetylation with the potential to impact mitochondrial metabolic and antioxidant processes., (Copyright © 2015. Published by Elsevier B.V.)
- Published
- 2015
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27. Does Continuous Hospice Care Help Patients Remain at Home?
- Author
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Casarett D, Harrold J, Harris PS, Bender L, Farrington S, Smither E, Ache K, and Teno J
- Subjects
- Aged, Caregivers, Cohort Studies, Death, Electronic Health Records, Family, Female, Hospitalization statistics & numerical data, Humans, Inpatients statistics & numerical data, Male, Nursing Homes statistics & numerical data, Home Care Services statistics & numerical data, Hospice Care methods, Hospice Care statistics & numerical data
- Abstract
Context: In the U. S., hospices sometimes provide high-intensity "continuous care" in patients' homes. However, little is known about the way that continuous care is used or what impact continuous care has on patient outcomes., Objectives: To describe patients who receive continuous care and determine whether continuous care reduces the likelihood that patients will die in an inpatient unit or hospital., Methods: Data from 147,137 patients admitted to 11 U.S. hospices between 2008 and 2012 were extracted from the electronic medical records. The hospices are part of a research-focused collaboration. The study used a propensity score-matched cohort design., Results: A total of 99,687 (67.8%) patients were in a private home or nursing home on the day before death, and of these, 10,140 (10.2%) received continuous care on the day before death. A propensity score-matched sample (n = 24,658) included 8524 patients who received continuous care and 16,134 patients who received routine care on the day before death. Using the two matched groups, patients who received continuous care on the day before death were significantly less likely to die in an inpatient hospice setting (350/8524 vs. 2030/16,134; 4.1% vs. 12.6%) (odds ratio [OR] 0.29; 95% CI 0.27-0.34; P < 0.001). When patients were cared for by a spouse, the use of continuous care was associated with a larger decrease in inpatient deaths (OR 0.12; 95% CI 0.09-0.16; P < 0.001) compared with those patients cared for by other family members (OR 0.37; 95% CI 0.32-0.42; P < 0.001). It is possible that unmeasured covariates were not included in the propensity score match., Conclusion: Use of continuous care on the day before death is associated with a significant reduction in the use of inpatient care on the last day of life, particularly when patients are cared for by a spouse., (Copyright © 2015. Published by Elsevier Inc.)
- Published
- 2015
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28. Hospice Care in Assisted Living Facilities Versus at Home: Results of a Multisite Cohort Study.
- Author
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Dougherty M, Harris PS, Teno J, Corcoran AM, Douglas C, Nelson J, Way D, Harrold JE, and Casarett DJ
- Subjects
- Aged, Aged, 80 and over, Cohort Studies, Female, Geriatric Assessment statistics & numerical data, Health Status, Hospices statistics & numerical data, Humans, Length of Stay statistics & numerical data, Male, Retrospective Studies, United States epidemiology, Assisted Living Facilities organization & administration, Home Care Services organization & administration, Hospice Care standards, Hospice Care statistics & numerical data, Independent Living statistics & numerical data
- Abstract
Objectives: To compare residents of assisted living facilities receiving hospice with people receiving hospice care at home., Design: Electronic health record-based retrospective cohort study., Setting: Nonprofit hospices in the Coalition of Hospices Organized to Investigate Comparative Effectiveness network., Participants: Individuals admitted to hospice between January 1, 2008, and May 15, 2012 (N = 85,581; 7,451 (8.7%) assisted living facility, 78,130 (91.3%) home)., Measurements: Hospice length of stay, use of opioids for pain, and site of death., Results: The assisted living population was more likely than the home hospice population to have a diagnosis of dementia (23.5% vs 4.7%; odds ratio (OR) = 13.3, 95% confidence interval (CI) = 12.3-14.4; P < .001) and enroll in hospice closer to death (median length of stay 24 vs 29 days). Assisted living residents were less likely to receive opioids for pain (18.1% vs 39.7%; OR = 0.33, 95% CI = 0.29-0.39, P < .001) and less likely to die in an inpatient hospice unit (9.3% vs 16.1%; OR = 0.53, 95% CI = 0.49-0.58, P < .001) or a hospital (1.3% vs 7.6%; OR = 0.16, 95% CI = 0.13-0.19, P < .001)., Conclusion: Three are several differences between residents of assisted living receiving hospice care and individuals living at home receiving hospice care. A better understanding of these differences could allow hospices to develop guidelines for better coordination of end-of-life care for the assisted living population., (© 2015, Copyright the Authors Journal compilation © 2015, The American Geriatrics Society.)
- Published
- 2015
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29. Hospice admissions for cancer in the final days of life: independent predictors and implications for quality measures.
- Author
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O'Connor NR, Hu R, Harris PS, Ache K, and Casarett DJ
- Subjects
- Aged, Female, Hematologic Neoplasms economics, Hematologic Neoplasms therapy, Hospice Care methods, Humans, Logistic Models, Male, Medicaid economics, Medicare economics, Middle Aged, Multivariate Analysis, Neoplasms therapy, Quality of Health Care economics, Referral and Consultation economics, Retrospective Studies, Time Factors, United States, Hospice Care economics, Hospitalization economics, Length of Stay economics, Neoplasms economics
- Abstract
Purpose: To define patient characteristics associated with hospice enrollment in the last 3 days of life, and to describe adjusted proportions of patients with late referrals among patient subgroups that could be considered patient-mix adjustment variables for this quality measure., Methods: Electronic health record-based retrospective cohort study of patients with cancer admitted to 12 hospices in the Coalition of Hospices Organized to Investigate Comparative Effectiveness network., Results: Of 64,264 patients admitted to hospice with cancer, 10,460 (16.3%) had a length of stay ≤ 3 days. There was significant variation among hospices (range, 11.4% to 24.5%). In multivariable analysis, among patients referred to hospice, patients who were admitted in the last 3 days of life were more likely to have a hematologic malignancy, were more likely to be male and married, and were younger (age < 65 years). Patients with Medicaid or self-insurance were less likely to be admitted to hospice within 3 days of death., Conclusion: Quality measures of hospice lengths of stay should include patient-mix adjustments for type of cancer and site of care. Patients with hematologic malignancies are at especially increased risk for late admission to hospice., (© 2014 by American Society of Clinical Oncology.)
- Published
- 2014
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30. Immediate and long-term consequences of vascular toxicity during zebrafish development.
- Author
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Tal TL, McCollum CW, Harris PS, Olin J, Kleinstreuer N, Wood CE, Hans C, Shah S, Merchant FA, Bondesson M, Knudsen TB, Padilla S, and Hemmer MJ
- Subjects
- Angiogenesis Inhibitors pharmacology, Animals, Animals, Genetically Modified, Blood Vessels drug effects, Embryo, Nonmammalian drug effects, Embryonic Development drug effects, Embryonic Development physiology, Phthalazines pharmacology, Protein Kinase Inhibitors pharmacology, Pyridines pharmacology, Quinazolines pharmacology, Tyrphostins pharmacology, Blood Vessels embryology, Embryo, Nonmammalian embryology, ErbB Receptors antagonists & inhibitors, Vascular Endothelial Growth Factor Receptor-2 antagonists & inhibitors, Zebrafish embryology
- Abstract
Proper formation of the vascular system is necessary for embryogenesis, and chemical disruption of vascular development may be a key event driving developmental toxicity. In order to test the effect of environmental chemicals on this critical process, we evaluated a quantitative assay in transgenic zebrafish using angiogenesis inhibitors that target VEGFR2 (PTK787) or EGFR (AG1478). Both PTK787 and AG1478 exposure impaired intersegmental vessel (ISV) sprouting, while AG1478 also produced caudal and pectoral fin defects at concentrations below those necessary to blunt ISV morphogenesis. The functional consequences of vessel toxicity during early development included decreased body length and survival in juvenile cohorts developmentally exposed to inhibitor concentrations sufficient to completely block ISV sprouting angiogenesis. These data show that concentration-dependent disruption of the presumed targets for these inhibitors produce adverse outcomes at advanced life stages., (Published by Elsevier Inc.)
- Published
- 2014
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- View/download PDF
31. Can hospices predict which patients will die within six months?
- Author
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Harris PS, Stalam T, Ache KA, Harrold JE, Craig T, Teno J, Smither E, Dougherty M, and Casarett D
- Subjects
- Adult, Aged, Aged, 80 and over, Disease Progression, Female, Humans, Length of Stay statistics & numerical data, Male, Middle Aged, Predictive Value of Tests, Probability, Retrospective Studies, Risk Factors, Sex Factors, Time Factors, Hospices, Mortality trends
- Abstract
Objective: To determine whether it is possible to predict, at the time of hospice enrollment, which patients will die within 6 months., Design: Electronic health record-based retrospective cohort study., Setting: Patients admitted to 10 hospices in the CHOICE network (Coalition of Hospices Organized to Investigate Comparative Effectiveness)., Participants: Hospice patients., Main Outcome Measures: Mortality at 6 months following hospice admission., Results: Among 126,620 patients admitted to 10 hospices, 118,532 (93.6%) died within 6 months. In a multivariable logistic regression model, five characteristics were independent predictors of 6-month mortality. For instance, patients younger than 65 years were less likely to die within 6 months (odds ratio [OR] 0.64; 95% confidence interval [CI] 0.45-0.91; p=0.014). Conversely, male patients were more likely to die within 6 months (OR 1.47; 95% CI 1.05-2.02; p=;0.036). After adjusting for other variables in this model, there were several subgroups with a low probability of 6-month probability (e.g., stroke and Palliative Performance Scale [PPS] score=50; adjusted probability of 6-month mortality=39.4%; 95% CI: 13.9%-72.5%). However, 95% confidence intervals of these 6-month mortality predictions extended above 50%., Conclusions: Hospices might use several variables to identify patients with a relatively low risk for 6-month mortality and who therefore may become ineligible to continue hospice services if they fail to show significant disease progression.
- Published
- 2014
- Full Text
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32. Integrated genomic analysis identifies the mitotic checkpoint kinase WEE1 as a novel therapeutic target in medulloblastoma.
- Author
-
Harris PS, Venkataraman S, Alimova I, Birks DK, Balakrishnan I, Cristiano B, Donson AM, Dubuc AM, Taylor MD, Foreman NK, Reigan P, and Vibhakar R
- Subjects
- Apoptosis drug effects, Cell Cycle genetics, Cell Cycle Proteins genetics, Cell Line, Tumor, Cell Proliferation drug effects, Child, Preschool, Gene Expression Regulation, Neoplastic drug effects, Genome, Human, Genomics, Humans, Medulloblastoma metabolism, Medulloblastoma pathology, Nuclear Proteins genetics, Protein Kinase Inhibitors administration & dosage, Protein-Tyrosine Kinases genetics, Pyrazoles administration & dosage, Pyrimidines administration & dosage, Pyrimidinones, Cell Cycle Proteins biosynthesis, Medulloblastoma genetics, Molecular Targeted Therapy, Nuclear Proteins biosynthesis, Protein-Tyrosine Kinases biosynthesis
- Abstract
Background: Medulloblastoma is the most common type of malignant brain tumor that afflicts children. Although recent advances in chemotherapy and radiation have improved outcomes, high-risk patients do poorly with significant morbidity., Methods: To identify new molecular targets, we performed an integrated genomic analysis using structural and functional methods. Gene expression profiling in 16 medulloblastoma patient samples and subsequent gene set enrichment analysis indicated that cell cycle-related kinases were associated with disease development. In addition a kinome-wide small interfering RNA (siRNA) screen was performed to identify kinases that, when inhibited, could prevent cell proliferation. The two genome-scale analyses were combined to identify key vulnerabilities in medulloblastoma. The inhibition of one of the identified targets was further investigated using RNAi and a small molecule inhibitor., Results: Combining the two analyses revealed that mitosis-related kinases were critical determinants of medulloblastoma cell proliferation. RNA interference (RNAi)-mediated knockdown of WEE1 kinase and other mitotic kinases was sufficient to reduce medulloblastoma cell proliferation. These data prompted us to examine the effects of inhibiting WEE1 by RNAi and by a small molecule inhibitor of WEE1, MK-1775, in medulloblastoma cell lines. MK-1775 inhibited the growth of medulloblastoma cell lines, induced apoptosis and increased DNA damage at nanomolar concentrations. Further, MK-1775 was synergistic with cisplatin in reducing medulloblastoma cell proliferation and resulted in an associated increase in cell death. In vivo MK-1775 suppressed medulloblastoma tumor growth as a single agent., Conclusions: Taken together, these findings highlight mitotic kinases and, in particular, WEE1 as a rational therapeutic target for medulloblastoma.
- Published
- 2014
- Full Text
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33. Survival after dialysis discontinuation and hospice enrollment for ESRD.
- Author
-
O'Connor NR, Dougherty M, Harris PS, and Casarett DJ
- Subjects
- Aged, Female, Hospital Mortality, Humans, Kidney Failure, Chronic diagnosis, Life Expectancy, Logistic Models, Male, Multivariate Analysis, Odds Ratio, Palliative Care, Prognosis, Proportional Hazards Models, Risk Factors, Time Factors, United States epidemiology, Hospices, Kidney Failure, Chronic mortality, Kidney Failure, Chronic therapy, Patient Admission, Renal Dialysis, Terminal Care, Withholding Treatment
- Abstract
Background and Objectives: Textbooks report that patients with ESRD survive for 7-10 days after discontinuation of dialysis. Studies describing actual survival are limited, however, and research has not defined patient characteristics that may be associated with longer or shorter survival times. The goals of this study were to determine the mean life expectancy of patients admitted to hospice after discontinuation of dialysis, and to identify independent predictors of survival time., Design, Setting, Participants, & Measurements: Data for demographics, clinical characteristics, and survival were obtained from 10 hospices for patients with ESRD who discontinued dialysis before hospice admission. Data were collected for patients admitted between January 1, 2008 and May 15, 2012. All hospices were members of the Coalition of Hospices Organized to Investigate Comparative Effectiveness network, which obtains de-identified data from an electronic medical record., Results: Of 1947 patients who discontinued dialysis, the mean survival after hospice enrollment was 7.4 days (range, 0-40 days). Patients who discontinued dialysis had significantly shorter survival compared with other patients (n=124,673) with nonrenal hospice diagnoses (mean survival 54.4 days; hazard ratio, 2.96; 95% confidence interval, 2.82 to 3.09; P<0.001). A Cox proportional hazards model identified seven independent predictors of earlier mortality after dialysis discontinuation, including male sex, referral from a hospital, lower functional status (Palliative Performance Scale score), and the presence of peripheral edema., Conclusions: Patients who discontinue dialysis have significantly shorter survival than other hospice patients. Individual survival time varies greatly, but several variables can be used to predict survival and tailor a patient's care plan based on estimated prognosis.
- Published
- 2013
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34. Inhibition of EZH2 suppresses self-renewal and induces radiation sensitivity in atypical rhabdoid teratoid tumor cells.
- Author
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Alimova I, Birks DK, Harris PS, Knipstein JA, Venkataraman S, Marquez VE, Foreman NK, and Vibhakar R
- Subjects
- Adolescent, Apoptosis, Biomarkers, Tumor metabolism, Blotting, Western, Cell Cycle, Cerebellum metabolism, Cerebellum pathology, Child, Enhancer of Zeste Homolog 2 Protein, Gene Expression Profiling, Humans, Immunoenzyme Techniques, Oligonucleotide Array Sequence Analysis, Polycomb Repressive Complex 2 antagonists & inhibitors, Polycomb Repressive Complex 2 genetics, RNA, Messenger genetics, RNA, Small Interfering genetics, Real-Time Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, Rhabdoid Tumor metabolism, Rhabdoid Tumor radiotherapy, Spheroids, Cellular, Teratoma metabolism, Teratoma radiotherapy, Tumor Cells, Cultured, Biomarkers, Tumor genetics, Cell Proliferation, Cellular Senescence, Polycomb Repressive Complex 2 metabolism, Radiation Tolerance, Rhabdoid Tumor pathology, Teratoma pathology
- Abstract
Introduction: Overexpression of the Polycomb repressive complex 2 (PRC2) subunit Enhancer of Zeste 2 (EZH2) occurs in several malignancies, including prostate cancer, breast cancer, medulloblastoma, and glioblastoma multiforme. Recent evidence suggests that EZH2 may also have a role in rhabdoid tumors. Atypical teratoid/rhabdoid tumor (ATRT) is a rare, high-grade embryonal brain tumor that occurs most commonly in young children and carries a very poor prognosis. ATRTs are characterized by absence of the chromatin remodeling protein SMARCB1. Given the role of EZH2 in regulating epigenetic changes, we investigated the role of EZH2 in ATRT., Methods: Microarray analysis was used to evaluate expression of EZH2 in ATRT tumor samples. We used shRNA and a chemical inhibitor of EZH2 to examine the impact of EZH2 inhibition on cell growth, proliferation, and tumor cell self-renewal., Results: Here, we show that targeted disruption of EZH2 by RNAi or pharmacologic inhibition strongly impairs ATRT cell growth, suppresses tumor cell self-renewal, induces apoptosis, and potently sensitizes these cells to radiation. Using functional analysis of transcription factor activity, we found the cyclin D1-E2F axis to be repressed after EZH2 depletion in ATRT cells., Conclusions: Our observations provide evidence that EZH2 disruption alters cell cycle progression and may be an important new therapeutic target, particularly in combination with radiation, in ATRT.
- Published
- 2013
- Full Text
- View/download PDF
35. MicroRNA 218 acts as a tumor suppressor by targeting multiple cancer phenotype-associated genes in medulloblastoma.
- Author
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Venkataraman S, Birks DK, Balakrishnan I, Alimova I, Harris PS, Patel PR, Handler MH, Dubuc A, Taylor MD, Foreman NK, and Vibhakar R
- Subjects
- 3' Untranslated Regions, Animals, Argonaute Proteins genetics, Argonaute Proteins metabolism, Carrier Proteins genetics, Carrier Proteins metabolism, Cathepsin B genetics, Cathepsin B metabolism, Cell Line, Tumor, Cell Movement, Cerebellar Neoplasms metabolism, Cerebellar Neoplasms pathology, Cerebellum pathology, Child, Preschool, Cyclin-Dependent Kinase 6 genetics, Cyclin-Dependent Kinase 6 metabolism, High-Throughput Nucleotide Sequencing, Humans, Medulloblastoma metabolism, Medulloblastoma pathology, Mice, MicroRNAs metabolism, Neoplasm Invasiveness, Neural Stem Cells metabolism, Neural Stem Cells pathology, Phenotype, RNA, Messenger biosynthesis, Rapamycin-Insensitive Companion of mTOR Protein, Repressor Proteins, Signal Transduction, Cerebellar Neoplasms genetics, Cerebellum metabolism, Gene Expression Regulation, Neoplastic, Medulloblastoma genetics, MicroRNAs genetics, RNA, Messenger antagonists & inhibitors
- Abstract
Aberrant expression of microRNAs has been implicated in many cancers. We recently demonstrated differential expression of several microRNAs in medulloblastoma. In this study, the regulation and function of microRNA 218 (miR-218), which is significantly underexpressed in medulloblastoma, was evaluated. Re-expression of miR-218 resulted in a significant decrease in medulloblastoma cell growth, cell colony formation, cell migration, invasion, and tumor sphere size. We used C17.2 neural stem cells as a model to show that increased miR-218 expression results in increased cell differentiation and also decreased malignant transformation when transfected with the oncogene REST. These results suggest that miR-218 acts as a tumor suppressor in medulloblastoma. MicroRNAs function by down-regulating translation of target mRNAs. Targets are determined by imperfect base pairing of the microRNA to the 3'-UTR of the mRNA. To comprehensively identify actual miR-218 targets, medulloblastoma cells overexpressing miR-218 and control cells were subjected to high throughput sequencing of RNA isolated by cross-linking immunoprecipitation, a technique that identifies the mRNAs bound to the RNA-induced silencing complex component protein Argonaute 2. High throughput sequencing of mRNAs identified 618 genes as targets of miR-218 and included both previously validated targets and many targets not predicted computationally. Additional work further confirmed CDK6, RICTOR, and CTSB (cathepsin B) as targets of miR-218 and examined the functional role of one of these targets, CDK6, in medulloblastoma.
- Published
- 2013
- Full Text
- View/download PDF
36. Inhibition of cyclin-dependent kinase 6 suppresses cell proliferation and enhances radiation sensitivity in medulloblastoma cells.
- Author
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Whiteway SL, Harris PS, Venkataraman S, Alimova I, Birks DK, Donson AM, Foreman NK, and Vibhakar R
- Subjects
- Animals, Cell Cycle drug effects, Cell Cycle radiation effects, Cell Differentiation drug effects, Cell Differentiation physiology, Cell Line, Tumor, Cell Proliferation drug effects, Cell Survival drug effects, Cell Survival radiation effects, Child, Preschool, Dose-Response Relationship, Drug, Dose-Response Relationship, Radiation, Enzyme Inhibitors pharmacology, Female, Gene Expression Regulation, Neoplastic drug effects, Gene Expression Regulation, Neoplastic radiation effects, Humans, Male, Medulloblastoma drug therapy, Mice, Neural Stem Cells drug effects, Neural Stem Cells physiology, Piperazines pharmacology, Pyridines pharmacology, RNA Interference physiology, Radiation Tolerance drug effects, Radiation Tolerance radiation effects, Radiation, Ionizing, Tubulin metabolism, Cerebellar Neoplasms pathology, Cyclin-Dependent Kinase 6 metabolism, Gene Expression Regulation, Neoplastic physiology, Medulloblastoma pathology, Radiation Tolerance physiology
- Abstract
Medulloblastoma accounts for 20 % of all primary pediatric intracranial tumors. Current treatment cures 50-80 % of patients but is associated with significant long-term morbidity and thus new therapeutic targets are needed. One such target is cyclin-dependent kinase 6 (CDK6), a serine/threonine kinase that plays a vital role in cell cycle progression and differentiation. CDK6 is overexpressed in medulloblastoma patients and is associated with an adverse prognosis. To investigate the role of CDK6 in medulloblastoma, we assayed the effect of CDK6 inhibition on proliferation by depleting expression with RNA interference (RNAi) or by inhibiting kinase function with a small molecule inhibitor, PD0332991. Cell proliferation was assessed by colony focus assay or by the xCELLigence system. We then investigated the impact of CDK6 inhibition on differentiation of murine neural stem cells by immunofluorescence of relevant markers. Finally we evaluated the effects of PD0332991 treatment on medulloblastoma cell cycle and radiosensitivity using colony focus assays. Gene expression analysis revealed that CDK6 mRNA expression is higher than normal cerebellum in fifteen out of sixteen medulloblastoma patient samples. Inhibition of CDK6 by RNAi significantly decreased medulloblastoma cell proliferation and colony forming potential. Interestingly, CDK6 inhibition by RNAi increased differentiation in murine neural stem cells. PD0332991 treatment significantly decreased medulloblastoma cell proliferation and led to a G0/G1 cell cycle arrest. Furthermore, PD0332991 pretreatment sensitized medulloblastoma cells to ionizing radiation. Our findings suggest that targeting CDK6 with small molecule inhibitors may prove beneficial in the treatment of medulloblastoma, especially when combined with radiation.
- Published
- 2013
- Full Text
- View/download PDF
37. Targeting Aurora Kinase A enhances radiation sensitivity of atypical teratoid rhabdoid tumor cells.
- Author
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Venkataraman S, Alimova I, Tello T, Harris PS, Knipstein JA, Donson AM, Foreman NK, Liu AK, and Vibhakar R
- Subjects
- Antineoplastic Agents pharmacology, Apoptosis drug effects, Apoptosis genetics, Aurora Kinase A, Aurora Kinases, Azepines pharmacology, Blotting, Western, Central Nervous System Neoplasms genetics, Enzyme Inhibitors pharmacology, Gene Expression Profiling, Humans, Oligonucleotide Array Sequence Analysis, Protein Serine-Threonine Kinases genetics, Pyrimidines pharmacology, Real-Time Polymerase Chain Reaction, Rhabdoid Tumor genetics, Teratoma genetics, Tumor Cells, Cultured, Central Nervous System Neoplasms enzymology, Protein Serine-Threonine Kinases metabolism, Radiation Tolerance genetics, Rhabdoid Tumor enzymology, Teratoma enzymology
- Abstract
Atypical teratoid/rhabdoid tumors (ATRT) are rare, highly malignant, embryonal CNS tumors with a poor prognosis. Therapy relies on highly toxic chemotherapy and radiotherapy. To improve outcomes and decrease morbidity, more targeted therapy is required. Gene expression analysis revealed elevated expression of multiple kinases in ATRT tissues. Aurora Kinase A was one of the candidate kinases. The objective of this study was to evaluate the impact of Aurora Kinase A inhibition in ATRT cell lines. Our analysis revealed that inhibition of Aurora Kinase A induces cell death in ATRT cells and the small molecule inhibitor MLN 8237 sensitizes these cells to radiation. Furthermore, inhibition of Aurora Kinase A resulted in decreased activity of pro-proliferative signaling pathways. These data indicate that inhibition of Aurora Kinase A is a promising small molecule target for ATRT therapy.
- Published
- 2012
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- View/download PDF
38. Polo-like kinase 1 (PLK1) inhibition suppresses cell growth and enhances radiation sensitivity in medulloblastoma cells.
- Author
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Harris PS, Venkataraman S, Alimova I, Birks DK, Donson AM, Knipstein J, Dubuc A, Taylor MD, Handler MH, Foreman NK, and Vibhakar R
- Subjects
- Apoptosis drug effects, Blotting, Western, Cell Cycle Proteins genetics, Cell Cycle Proteins metabolism, Cell Line, Tumor, Cell Proliferation drug effects, Cerebellar Neoplasms enzymology, Cerebellar Neoplasms pathology, Child, Child, Preschool, Cohort Studies, Enzyme Inhibitors pharmacology, Female, Humans, Male, Medulloblastoma enzymology, Medulloblastoma pathology, Microarray Analysis, Neoplasm Proteins genetics, Neoplasm Proteins metabolism, Protein Serine-Threonine Kinases genetics, Protein Serine-Threonine Kinases metabolism, Proto-Oncogene Proteins genetics, Proto-Oncogene Proteins metabolism, Pteridines pharmacology, RNA metabolism, RNA, Mitochondrial, Polo-Like Kinase 1, Cell Cycle Proteins antagonists & inhibitors, Cerebellar Neoplasms radiotherapy, Medulloblastoma radiotherapy, Neoplasm Proteins antagonists & inhibitors, Protein Serine-Threonine Kinases antagonists & inhibitors, Proto-Oncogene Proteins antagonists & inhibitors, Radiation Tolerance drug effects
- Abstract
Background: Medulloblastoma is the most common malignant brain tumor in children and remains a therapeutic challenge due to its significant therapy-related morbidity. Polo-like kinase 1 (PLK1) is highly expressed in many cancers and regulates critical steps in mitotic progression. Recent studies suggest that targeting PLK1 with small molecule inhibitors is a promising approach to tumor therapy., Methods: We examined the expression of PLK1 mRNA in medulloblastoma tumor samples using microarray analysis. The impact of PLK1 on cell proliferation was evaluated by depleting expression with RNA interference (RNAi) or by inhibiting function with the small molecule inhibitor BI 2536. Colony formation studies were performed to examine the impact of BI 2536 on medulloblastoma cell radiosensitivity. In addition, the impact of depleting PLK1 mRNA on tumor-initiating cells was evaluated using tumor sphere assays., Results: Analysis of gene expression in two independent cohorts revealed that PLK1 mRNA is overexpressed in some, but not all, medulloblastoma patient samples when compared to normal cerebellum. Inhibition of PLK1 by RNAi significantly decreased medulloblastoma cell proliferation and clonogenic potential and increased cell apoptosis. Similarly, a low nanomolar concentration of BI 2536, a small molecule inhibitor of PLK1, potently inhibited cell growth, strongly suppressed the colony-forming ability, and increased cellular apoptosis of medulloblastoma cells. Furthermore, BI 2536 pretreatment sensitized medulloblastoma cells to ionizing radiation. Inhibition of PLK1 impaired tumor sphere formation of medulloblastoma cells and decreased the expression of SRY (sex determining region Y)-box 2 (SOX2) mRNA in tumor spheres indicating a possible role in targeting tumor initiating cells., Conclusions: Our data suggest that targeting PLK1 with small molecule inhibitors, in combination with radiation therapy, is a novel strategy in the treatment of medulloblastoma that warrants further investigation.
- Published
- 2012
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- View/download PDF
39. Application of protein expression profiling to screen chemicals for androgenic activity.
- Author
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Hemmer MJ, Salinas KA, and Harris PS
- Subjects
- Animals, Biomarkers blood, Dihydrotestosterone toxicity, Endosulfan toxicity, Female, Fresh Water chemistry, Insecticides toxicity, Methyltestosterone toxicity, Testosterone toxicity, Trenbolone Acetate toxicity, Androgens toxicity, Cyprinidae blood, Fish Proteins blood, Water Pollutants, Chemical toxicity
- Abstract
Protein expression changes can be used for detection of biomarkers that can be applied diagnostically to screen chemicals for endocrine modifying activity. In this study, surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) coupled with a short term fish assay was used to investigate changes in plasma protein expression as a means to screen chemicals for androgenic activity. Adult gravid female sheepshead minnows (Cyprinodon variegatus) were placed into separate aquaria for seawater control, ethanol solvent control, and the following androgen agonist treatments at 5.0μg/L: dihydrotestosterone (DHT), methyldihydrotestosterone (MDHT), testosterone (T), methyltestosterone (MT) and trenbolone (TB). Treatments of 0.6μg/L endosulfan and 40μg/L chlorpyrifos (CP) served as non-androgenic negative stressor controls. Test concentrations were maintained using an intermittent flow-through dosing apparatus supplying exposure water at 20L/h. Fish were sampled at 7 days, the plasma diluted, processed on weak cation exchange CM10 ProteinChip arrays and analyzed. Spectral processing resulted in 249 individual m/z peak clusters for the androgen exposed fish. Partial least squares-discriminant analysis was used to develop an androgen-responsive model using sample spectra from exposures with DHT and unexposed solvent control fish as the training set. The androgen classification model performed with ≥79% specificity (% true negative) and ≥70% sensitivity (% true positive) for non-aromatizable androgens. The aromatizable androgens T and MT were classified as androgenic with specificities of 42 and 79%, respectively. The reduction in sensitivity observed with T is thought to be caused by its metabolic conversion to an estrogen by aromatase. The results of these studies show diagnostic plasma protein expression models can correctly classify chemicals by their androgenic activity using a combination of high throughput mass spectrometry and multivariate approaches., (Published by Elsevier B.V.)
- Published
- 2011
- Full Text
- View/download PDF
40. Caspase inhibition blocks cell death and enhances mitophagy but fails to promote T-cell lymphoma.
- Author
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Wang SH, Martin SM, Harris PS, and Knudson CM
- Subjects
- Animals, Apoptosis Regulatory Proteins genetics, Beclin-1, Caspases metabolism, Cell Death drug effects, Cell Proliferation drug effects, Cell Separation, Cells, Cultured, Dexamethasone pharmacology, Gene Dosage genetics, Genes, Dominant, Haploinsufficiency genetics, Mice, Mice, Transgenic, Mitochondria drug effects, Mitochondria ultrastructure, Reactive Oxygen Species metabolism, Thymus Gland pathology, bcl-2-Associated X Protein metabolism, Autophagy drug effects, Caspase Inhibitors, Lymphoma, T-Cell enzymology, Lymphoma, T-Cell pathology, Mitochondria pathology
- Abstract
Caspase-9 is a component of the apoptosome that mediates cell death following release of cytochrome c from mitochondria. Inhibition of Caspase-9 with a dominant negative construct (Casp9DN) blocks apoptosome function, promotes viability and has been implicated in carcinogenesis. Inhibition of the apoptosome in vitro impairs mitochondrial function and promotes mitophagy. To examine whether inhibition of the apoptosome would enhance mitophagy and promote oncogenesis in vivo, transgenic mice were generated that express Casp9DN in the T cell lineage. The effects of Casp9DN on thymocyte viability, mitophagy and thymic tumor formation were examined. In primary thymocytes, Casp9DN delayed dexamethasone (Dex)-induced cell death, altered mitochondrial structure, and decreased oxidant production. Transmission electron microscopy (TEM) revealed that inhibition of the apoptosome resulted in structurally abnormal mitochondria that in some cases were engulfed by double-membrane structures resembling autophagosomes. Consistent with mitochondria being engulfed by autophagosomes (mitophagy), confocal microscopy showed colocalization of LC3-GFP and mitochondria. However, Casp9DN did not significantly accelerate T-cell lymphoma alone, or in combination with Lck-Bax38/1, or with Beclin 1+/- mice, two tumor-prone strains in which altered mitochondrial function has been implicated in promoting tumor development. In addition, heterozygous disruption of Beclin 1 had no effect on T-cell lymphoma formation in Lck-Bax38/1 mice. Further studies showed that Beclin 1 levels had no effect on Casp9DN-induced loss of mitochondrial function. These results demonstrate that neither inhibition of apoptosome function nor Beclin 1 haploinsufficiency accelerate T-cell lymphoma development in mice.
- Published
- 2011
- Full Text
- View/download PDF
41. Urinary excretion of radionuclides from Marshallese exposed to fallout from the 1954 Bravo nuclear test.
- Author
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Harris PS, Simon SL, and Ibrahim SA
- Subjects
- Adult, Biological Assay, Child, Geography, Humans, Iodine Radioisotopes adverse effects, Iodine Radioisotopes metabolism, Iodine Radioisotopes urine, Micronesia epidemiology, Radiation Dosage, Radioactive Fallout adverse effects, Radioisotopes adverse effects, Radioisotopes metabolism, Risk Assessment, Time Factors, Nuclear Weapons, Radiation Monitoring, Radioactive Fallout analysis, Radioisotopes urine
- Abstract
Soon after the Bravo nuclear test at Bikini Atoll in the Marshall Islands on 1 March 1954, urine samples were collected for analysis of excreted radioactivity from native residents exposed to radioactive fallout on two atolls as well as from U.S. military personnel on a third atoll. The earliest acquired samples, obtained by the Los Alamos Scientific Laboratory (LASL), were assayed for various radionuclides and provided the first known measurements of (131)I in urine following exposure to fallout from a nuclear test. Over the course of 1954, many additional samples were collected by the LASL, as well as by the Atomic Energy Commission New York Operations Office's Health and Safety Laboratory and the Naval Radiological Defense Laboratory. Collectively, the groups sampled included Marshallese exposed on Rongelap and Ailinginae Atolls, American military weather observers temporarily resident on Rongerik Atoll, and sailors from the Japanese fishing vessel, the Lucky Dragon. While the bioassay measurement data and individual urine volumes have been crucial to various attempts to assess intakes of radioactivity and the related internal radiation doses among the Marshallese, those data have never been published in any peer-reviewed journal, but have been restricted to agency memoranda, laboratory reports, and summaries in some publications and book chapters. Reconstructions of internal doses to Marshallese in 1954 and in later years have depended on these data and, hence, they have considerable historical importance as well as importance to ongoing health risk projections for Marshallese. This paper presents much of the original data on urine volumes and radioactivity from the various assays of urine for radionuclides, and compares estimates of (131)I intakes made in 1954, 1985, 1987, and 2008.
- Published
- 2010
- Full Text
- View/download PDF
42. A simple and rapid matrix-assisted laser desorption/ionization time of flight mass spectrometry method to screen fish plasma samples for estrogen-responsive biomarkers.
- Author
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Salinas KA, Hemmer MJ, Harris PS, and Walker CC
- Subjects
- Animals, Estrogen Receptor Modulators administration & dosage, Male, Biomarkers blood, Estrogens blood, Fishes blood, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization methods
- Abstract
In the present study, we describe and evaluate the performance of a simple and rapid mass spectral method for screening fish plasma for estrogen-responsive biomarkers using matrix-assisted laser desorption/ionization (MALDI) time of flight mass spectrometry coupled with a short-term fish assay. Adult male sheepshead minnows (Cyprinodon variegatus) were placed into aquaria consisting of vehicle control and the following estrogen agonist treatments: 17beta-estradiol (0.00625, 0.0125, 0.025, 0.05, 0.1, 0.2, 0.5, and 1.0 microg/L, 4-tert-pentylphenol (100 microg/L), methoxychlor (6 and 12 microg/L), and bisphenol A (100 and 1,000 microg/L). Treatments with chlorpyrifos (80 microg/L) and endosulfan (0.6 microg/L) served as nonestrogenic negative controls. Test concentrations were maintained using an intermittent flow-through dosing apparatus. Plasma was obtained from individuals, diluted and applied to an inert surface, and analyzed by MALDI. Multiple protein peaks, ranging from 2.9 to 12.9 kDa, were identified as markers of estrogenic effects when comparing estrogen-treated and control fish using interpercentile reference values. A binary classification tree model was constructed from plasma protein profiles of the vehicle control and the 0.2 microg/L of 17beta-estradiol treatments and then used to evaluate all samples. Treatments with the estrogen agonists 17beta-estradiol, 4-tert-pentylphenol, methoxychlor, and bisphenol-A generated reproducible diagnostic biomarkers based on the presence of specific estrogen-responsive plasma proteins. The controls and nonestrogenic compounds chlorpyrifos and endosulfan did not produce this estrogen-responsive protein profile. A no-observed-effect level for 17beta-estradiol at 0.025 microg/L was estimated from concentration-response exposures. The MALDI method described here provides a straightforward, sensitive, and specific tool to screen chemicals for estrogenic activity.
- Published
- 2008
- Full Text
- View/download PDF
43. Characterizing coral condition using estimates of three-dimensional colony surface area.
- Author
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Fisher WS, Davis WP, Quarles RL, Patrick J, Campbell JG, Harris PS, Hemmer BL, and Parsons M
- Subjects
- Animals, Population Density, Population Dynamics, Anthozoa physiology, Biodiversity, Ecosystem, Environmental Monitoring methods
- Abstract
Coral reefs provide shoreline protection, biological diversity, fishery harvests, and tourism, all values that stem from the physically-complex coral infrastructure. Stony corals (scleractinians) construct and maintain the reef through deposition of calcium carbonate. Therefore, assessment of coral reefs requires at least some measurement endpoints that reflect the biological and physical condition of stony corals. Most monitoring programs portray coral quantity as live coral cover, which is the two-dimensional proportion of coral surface to sea floor viewed from above (planar view). The absence of the third dimension, however, limits our ability to characterize coral reef value, physiology, health and sustainability. A three-dimensional (3D) approach more realistically characterizes coral structure available as community habitat and, when combined with estimates of live coral tissue, quantifies the amount of living coral available for photosynthesis, growth and reproduction. A rapid coral survey procedure that coupled 3D coral quantification with more traditional survey measurements was developed and tested in the field. The survey procedure relied on only three underwater observations--species identification, colony size, and proportion of live tissue--made on each colony in the transect. These observations generated a variety of metrics, including several based on 3D colony surface area, that are relevant to reef management.
- Published
- 2007
- Full Text
- View/download PDF
44. A proteomic (SELDI-TOF-MS) approach to estrogen agonist screening.
- Author
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Walker CC, Salinas KA, Harris PS, Wilkinson SS, Watts JD, and Hemmer MJ
- Subjects
- Animals, Benzhydryl Compounds, Biomarkers blood, Chlorpyrifos toxicity, Dose-Response Relationship, Drug, Egg Proteins blood, Endosulfan toxicity, Environmental Monitoring methods, Estradiol toxicity, Female, Male, Methoxychlor toxicity, Phenols toxicity, Protein Array Analysis, Reproducibility of Results, Sensitivity and Specificity, Structure-Activity Relationship, Cyprinidae blood, Estrogens toxicity, Fish Proteins blood, Proteomics methods, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Water Pollutants, Chemical toxicity
- Abstract
A small fish model and surface-enhanced laser desorption/ionization time-of-flight mass spectrometry were used to investigate plasma protein expression as a means to screen chemicals for estrogenic activity. Adult male sheepshead minnows (Cyprinodon variegatus) were placed into aquaria for seawater control, solvent control, and treatments of 17beta-estradiol (E2), methoxychlor (MXC), bisphenol-A (BPA), 4-tert-pentylphenol (TPP), endosulfan (ES), and chlorpyriphos (CP). Fish plasma was applied to weak cation exchange (CM10) ProteinChip arrays, processed, and analyzed. The array produced approximately 42 peaks for E2 plasma and 30 peaks for solvent control plasma. Estrogen-responsive mass spectral biomarker peaks were identified by comparison of E2-treated and control plasma spectra. Thirteen potential protein biomarkers with a range from 1 to 13 kDa were up- or downregulated in E2-treated fish and their performance as estrogenic effects markers was evaluated by comparing spectra from control, estrogen agonist, and nonagonist stressor-treated males and normal female fish plasma. One of the biomarkers, mass-to-charge ratio 3025.5, was identified by high-resolution tandem mass spectrometry as C. variegatus zona radiata protein, fragment 2. The weak environmental estrogens MXC, BPA, and TPP elicited protein expression profiles consistent with the estrogen expression model. Estrogen-responsive peaks were not detected in plasma from fish in the seawater, vehicle, ES, or CP treatments. No difference was found between plasma protein expression of seawater control and solvent control fish. We show that water exposure of fish to estrogen agonists produces distinct plasma protein biomarkers that can be reproducibly detected at low levels using protein chips and mass spectrometry.
- Published
- 2007
- Full Text
- View/download PDF
45. Effects of fenoxycarb exposure on complete larval development of the xanthid crab, Rhithropanopeus harrisii.
- Author
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Cripe GM, McKenney CL Jr, Hoglund MD, and Harris PS
- Subjects
- Animals, Brachyura growth & development, Dose-Response Relationship, Drug, Larva drug effects, Metamorphosis, Biological drug effects, Seawater chemistry, Brachyura drug effects, Carbamates toxicity, Insecticides toxicity, Phenylcarbamates, Water Pollutants, Chemical toxicity
- Abstract
Pest control agents, such as juvenile hormone analogues (JHA), have been developed to limit effects on non-target organisms that co-inhabit insect pest habitats. Rhithropanopeus harrisii, an estuarine xanthid crab, was used to observe the impacts of the JHA, fenoxycarb, on the pattern of complete larval development as well as survival of larvae and successful metamorphosis to first crab stage. Significant mortality occurred in the first of four zoeal stages (after 2-3 days of exposure) at the highest treatment of 240 microg fenoxycarb/l and in megalopae exposed to 48 microg fenoxycarb/l. The time required to metamorphose to the first crab stage was significantly increased for megalopae in all treatments 48 microg/l. This delay in development was sufficient to significantly prolong the entire developmental period from zoea to crabs. Unexposed larvae developed to crabs in an average of 16 days; larvae exposed to >/=48 microg/l required 19-20 days. Reduced survival and extended duration of developing larval stages in the life history of a benthic invertebrate may alter the population dynamics of these organisms in the estuary.
- Published
- 2003
- Full Text
- View/download PDF
46. Sediment chemical contamination and toxicity associated with a coastal golf course complex.
- Author
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Lewis MA, Foss SS, Harris PS, Stanley RS, and Moore JC
- Subjects
- Animals, Environmental Monitoring, Environmental Pollutants analysis, Golf, Toxicity Tests, Water Movements, Environmental Pollutants toxicity, Geologic Sediments chemistry, Invertebrates, Metals, Heavy analysis, Pesticides analysis, Soil Pollutants analysis
- Abstract
The increasing density of golf courses represents a potential source of sediment contamination to nearby coastal areas, the chemical and biological magnitude of which is almost unknown. The objective of this study was to determine the concentrations of contaminants and toxicities of sediments impacted by a coastal golf course complex. Sediment contaminant concentrations were determined at least twice during the two-year study period at 14 sampling stations. In addition, a combination of acute and chronic bioassays were conducted exposing four invertebrate test species to whole sediments and associated pore waters. Overall, the Florida, USA, golf course complex had a measurable impact on sediment chemical quality, particularly in near-field areas. Higher concentrations of several trace metals and organochlorine pesticides were detected in many golf course-associated sediments compared with reference areas; however, concentrations decreased seaward and only a few, primarily chlorinated pesticides, exceeded proposed sediment quality guidelines. Chromium, zinc, and mercury were detected more frequently than other trace metals. The DDT and associated metabolites, dieldrin and chlordane, were the more commonly detected organic contaminants. Acute toxicity was uncommon and occurred consistently for sediment collected from one coastal location. In contrast, chronic toxicity occurred at several study sites based on the response of Mysidopsis bahia. It was concluded that the impact of golf course runoff on sediment quality may be subtle and sensitive biological assessment methods, such as chronic toxicity tests, will be needed to detect adverse effects.
- Published
- 2001
- Full Text
- View/download PDF
47. Effects of whole sediments from Corpus Christi Bay on survival, growth, and reproduction of the mysid, Americamysis bahia (formerly Mysidopsis bahia).
- Author
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Cripe GM, Carr RS, Foss SS, Harris PS, and Stanley RS
- Subjects
- Animals, Decapoda growth & development, Geologic Sediments, Mortality, Survival Analysis, Decapoda drug effects, Reproduction, Water Pollutants, Chemical adverse effects
- Published
- 2000
- Full Text
- View/download PDF
48. The "impossible patient": organizational response to a clinical problem.
- Author
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Harris PS, Duermeyer M, Ehly C, Hartig-Toth S, Hayes S, Holsapple L, and Peters D
- Subjects
- Adult, Ethical Analysis, Ethics Committees, Ethics Committees, Clinical, Ethics, Clinical, Humans, Kansas, Male, Patient Care Planning, Personal Autonomy, Rehabilitation Centers, Risk Management, Behavior Control, Contracts, Ethics, Medical, Health Care Sector, Physician-Patient Relations, Treatment Refusal
- Published
- 1999
49. Cardiac, cerebral, and vascular complications of infective endocarditis.
- Author
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Harris PS and Cobbs CG
- Subjects
- Brain Diseases classification, Heart Diseases classification, Humans, Vascular Diseases classification, Brain Diseases etiology, Endocarditis, Bacterial complications, Heart Diseases etiology, Vascular Diseases etiology
- Abstract
The complications of IE may involve any organ system. Cardiac complications are frequently present, and heart failure remains a leading cause of death. Abscess formation in the surrounding cardiac tissues may result in myocardial or pericardial disease, and cardiac conduction abnormalities may develop. Extracardiac complications, including neurologic, vascular, and renal diseases, are also common and are usually caused by either embolization of vegetations or deposition of immune complexes. Despite many advancements in the detection and treatment of the complications of IE, management of these problems remains a challenging endeavor.
- Published
- 1996
- Full Text
- View/download PDF
50. Modulation of hepatic protein kinase activity by indomethacin.
- Author
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Ganguli S, Sterman B, Harris PS, Sinha MK, and Banach WJ
- Subjects
- Adenylyl Cyclases metabolism, Animals, Binding Sites drug effects, Carrier Proteins metabolism, Cell Membrane enzymology, Cyclic AMP biosynthesis, Glucagon metabolism, Glucagon pharmacology, Liver drug effects, Male, Rats, Rats, Inbred Strains, Cyclic AMP Receptor Protein, Indomethacin pharmacology, Intracellular Signaling Peptides and Proteins, Liver enzymology, Protein Kinases metabolism
- Abstract
We previously demonstrated that treatment with indomethacin in vivo significantly blunted the glucagon-induced glycemic response in the rat. This prostaglandin synthetase (cyclo-oxygenase) inhibitor also accentuated the evanescent effect of glucagon on hepatic glucose output in the intact, anesthetized rat. In this report, we present evidence that impairment of glucagon action in the rat liver by indomethacin is mediated through its inhibitory effect on both cAMP-dependent and cAMP-independent hepatic protein kinase. Indomethacin treatment did not have a measurable effect on any of the other components of the glucagon transducer system. Furthermore, infusion with glucagon for two hours that maintained plasma glucagon values at high physiological levels significantly reduced hepatic cAMP-dependent protein kinase activity without altering its Km. Glucagon infusion also down-regulated its own hepatic receptors and glucagon-stimulated cAMP production; prostaglandin E1-stimulated cAMP production was not affected. We concluded that prostaglandins may play a role in the regulation of hepatic protein kinases involved in the glucagon-stimulated glycogenolytic response and that glucagon-induced down-regulation extends at least to the hepatic protein kinases. However, a direct effect of indomethacin or protein kinase and the adenylate cyclase complex cannot be ruled out.
- Published
- 1984
- Full Text
- View/download PDF
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