DOCSLIB.ORG

Neurometabolic Implications of Coenzyme Q10 Deficiency: Pathogenesis, Detection and Treatment 1

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

Neurometabolic Implications of Coenzyme Q10 Deficiency: Pathogenesis, Detection and Treatment Submitted By: Kate Duberley Department of Molecular Neuroscience UCL Institute of Neurology Queen Square, London Submitted July 2013 Funded by Ataxia UK Thesis submitted for the degree of Doctor of Philosophy, University College London (UCL) 1 I, Kate Duberley confirm that the work presented in this thesis is my own. Where information has been derived from other sources, I confirm that this has been indicated in the thesis. Signed………………………………………………….Date…………………… 2 Abstract Disorders of Coenzyme Q10 (CoQ10) biosynthesis represent the most treatable subgroup of mitochondrial diseases. Neurological involvement is frequently observed in CoQ10 deficiency, typically presenting as cerebellar ataxia and/or seizures. The aetiology of the neurological presentation of CoQ10 deficiency has yet to be fully elucidated and therefore in order to investigate these phenomena we have established a neuronal cell model of CoQ10 deficiency by treatment of the neuronal SH-SY5Y cell line with Para-AminoBenzoic Acid (PABA). This neuronal cell model provides insights into the effects of CoQ10 deficiency on neuronal mitochondrial function and oxidative stress. A marginal decrease in CoQ10 status (76% residual CoQ10) appears to be sufficient to impair Electron Transport Chain (ETC) function and increase mitochondrial oxidative stress, highlighting the vulnerability of neurons to a small deficit in CoQ10 status. In contrast to CoQ10 deficient fibroblasts, a CoQ10 deficiency (46% residual CoQ10) in neuronal cells appears to result in reversal of Complex V activity. This phenomenon has not been reported in previous studies of CoQ10 deficiency and may be a unique characteristic of neuronal cells. This neuronal cell model was subsequently utilised in the evaluation of candidate therapies for neurological conditions associated with CoQ10 deficiency. The efficacy of CoQ10 supplementation and methylene blue (MB) treatment were evaluated. CoQ10 supplementation proved effective at preventing mitochondrial oxidative stress and partially restoring neuronal mitochondrial function. However ETC complex activities were still compromised, suggesting an explanation for the refractory nature of neurological CoQ10 deficiency to treatment. Muscle is considered the “gold standard” for CoQ10 quantification; however neurological CoQ10 deficiency does not always present with a significant decrease in muscle CoQ10 status, despite a genetic diagnosis of CoQ10 deficiency. Cerebrospinal Fluid (CSF) CoQ10 quantification offers a more direct measurement of cerebellar CoQ10 levels. A tandem mass spectrometry (MS/MS) method capable of quantifying nanomolar (nM) levels of CoQ10 was therefore developed. 3 In conclusion this PhD thesis has been successful in expanding our understanding of the pathophysiology of neuronal CoQ10 deficiency and subsequently suggesting why neurological CoQ10 deficiency might be refractory to CoQ10 treatment. This thesis has also led to the development of a new technique for quantification of CSF CoQ10 concentration, opening up many possibilities for future studies and applications. This project is funded by Ataxia UK (www.ataxiauk.org.uk). 4 Table of Contents NEUROMETABOLIC IMPLICATIONS OF COENZYME Q10 DEFICIENCY: PATHOGENESIS, DETECTION AND TREATMENT 1 ABSTRACT 3 TABLE OF CONTENTS 5 TABLE OF FIGURES 11 LIST OF TABLES 15 LIST OF ABBREVIATIONS 16 ACKNOWLEDGEMENTS 19 CHAPTER 1 21 INTRODUCTION 21 1.1. COENZYME Q10 STRUCTURE 22 1.2. COENZYME Q10 FUNCTION 24 1.2.1. The Mitochondrial Electron Transport Chain (ETC) 24 1.2.2. Mitochondrial Oxidative Stress and the Antioxidant Role of Coenzyme Q10 36 1.2.3. The Plasma Membrane Redox System 41 1.2.4. Lysosomal Electron Transfer 41 1.2.5. The Mitochondrial Permeability Transition Pore (PTP) 42 1.2.6. Uncoupling Proteins (UCPs) 42 1.2.7. Cofactor in Pyrimidine Biosynthesis 43 1.2.8. Regulation of Cellular Membranes 43 1.3. COQ BIOSYNTHESIS 44 1.3.1. The Mevalonate Pathway 44 1.3.2. CoQ Biosynthesis 45 1.4. HUMAN COQ10 DEFICIENCY 49 5 1.4.1. COQ2 (encoding PHB-polyprenyl transferase) 50 1.4.2. PDSS1 and 2 (encoding transprenyl transferase) 51 1.4.3. ADCK3 (or CABC1) 51 1.4.4. COQ9 52 1.4.5. COQ6 52 1.4.6. COQ4 53 1.4.7. Secondary CoQ10 Deficiency 55 1.5. COQ10 SUPPLEMENTATION 58 1.5.1. Primary CoQ10 Deficiency 59 1.5.2. Secondary CoQ10 Deficiency 60 1.6. AIMS 61 CHAPTER 2 62 MATERIALS AND METHODS 62 2.1. MATERIALS 63 2.2. TISSUE CULTURE 64 2.2.1. Cell Line 64 2.2.2. Cell Passage 64 2.2.3. Cell Storage and Recovery 66 2.2.4. Treatment of SH-SY5Y Cells 66 2.2.5. SH-SY5Y Cell Harvesting for Biochemical Analysis 67 2.2.6. SH-SY5Y Cell Harvesting for Microscopy 67 2.3. TOTAL PROTEIN DETERMINATION 70 2.4. COENZYME Q10 QUANTIFICATION 70 2.4.1. Equipment 70 2.4.2. Procedure 70 2.4.3. Data Analysis 71 2.5. REDUCED GLUTATHIONE (GSH) QUANTIFICATION 74 6 2.5.1. Equipment 74 2.5.2. Procedure 74 2.5.3. Data Analysis 75 2.6. ENZYME ASSAYS 77 2.6.1. Complex I Assay (NADH:ubiquinone reductase; EC 1.6.5.3) 77 2.6.2. Complex II/III Assay (succinate cytochrome c reductase; EC 1.8.1.3) 78 2.6.3. Complex IV Assay (Cytochrome c oxidase; EC 1.9.3.1) 78 2.6.4. Citrate Synthase Assay (CS; EC 4.1.3.7) 79 2.6.5. Complex V (EC 3.6.3.14) in Gel Assay 82 2.6.6. Adenosine Triphospate (ATP) Assay 83 2.7. CONFOCAL MICROSCOPY 86 2.7.1. Preparation of Cells 86 2.7.2. Confocal microscopy method 86 2.7.3. Mitochondrial Membrane Potential 86 2.7.4. Mitochondrial Superoxide 88 2.7.5. Cytosolic Superoxide 88 2.7.6. Lipid Peroxidation 91 2.7.7. Mitochondrial DNA Quantification 91 2.8. STATISTICAL ANALYSIS 92 CHAPTER 3 95 NEURONAL CELL MODEL OF COENZYME Q10 DEFICIENCY 95 3.1. INTRODUCTION 96 3.2. AIMS 97 3.3. METHODS 98 3.3.1. Cell Culture 98 3.3.2. Quantification of CoQ10 Levels 98 7 3.3.3. Mitochondrial Bioenergetics Analysis 98 3.3.4. Blue Native in-Gel Complex V Assay 98 3.3.5. Oxidative Stress Assessment 98 3.3.6. Mitochondrial DNA Quantification 98 3.3.7. Mitochondrial Membrane Potential 98 3.3.8. Total Protein Determination 98 3.3.9. Statistical Analysis 98 3.4. RESULTS 99 3.4.1. Optimisation of PABA Treatment 99 3.4.2. Electron Transport Chain Activities 101 3.4.3. Cellular ATP Status 103 3.4.4. Blue Native Complex V Assay 103 3.4.5. Oxidative Stress Analysis 105 3.4.6. Mitochondrial DNA Quantification 106 3.4.7. Mitochondrial Membrane Potential 106 3.4.8. Co-incubation of PABA and CoQ10 109 3.5. DISCUSSION 112 3.6. CONCLUSION 116 CHAPTER 4 117 INVESTIGATION OF COENZYME Q10 AND METHYLENE BLUE TREATMENT ON THE NEURONAL CELL MODEL OF COENZYME Q10 DEFICIENCY 117 4.1. INTRODUCTION 118 4.2. AIMS 121 4.3. METHODS 121 4.3.1. Cell Culture 121 4.3.2. Quantification of CoQ10 Levels 121 4.3.3. Mitochondrial Bioenergetics Analysis 121 8 4.3.4. Oxidative Stress Assessment 121 4.3.5. Mitochondrial Membrane Potential 121 4.3.6. Total Protein Determination 122 4.3.7. Statistical Analysis 122 4.4. RESULTS 122 4.4.1. Treatment Optimisation 122 4.4.2. Coenzyme Q10 Treated Cells 124 4.4.3. Methylene Blue Treated Cells 128 4.5. DISCUSSION 131 4.6. CONCLUSION 134 CHAPTER 5 136 TANDEM MASS SPECTROMETRY METHOD FOR QUANTIFYING COENZYME Q10 IN CSF 136 5.1. BACKGROUND 137 5.2. AIMS 138 5.3. SYNTHESIS OF DEUTERATED COQ10 (D6-COQ10) 138 5.4. TANDEM MASS SPECTROMETRY METHOD DEVELOPMENT 139 5.4.1. Preparation of Standard Solutions 139 5.4.2. Subjects 140 5.4.3. Preparation of Patient Samples 140 5.4.4. Electrospray Ionisation-MS/MS (ESI-MS/MS) 144 5.4.5. High Performance Liquid Chromatography (HPLC) 147 5.5. RESULTS 154 5.5.1. Deuterated Internal Standard 154 5.5.2. Linearity, Precision, Recovery 155 5.5.3. Muscle Analysis 156 5.5.4. Fibroblast and CSF Analysis 157 9 5.6. DISCUSSION 159 5.7. CONCLUSION 162 CHAPTER 6 164 GENERAL DISCUSSION 164 6.1. FUTURE WORK 171 6.1.1. Neuronal Cell Model 171 6.1.2. Treatments 171 6.1.3. MS Method 172 6.2. REFERENCES 173 10 Table of Figures Figure 1.1 The Chemical Structure of Coenzyme Q10 (CoQ10)..........................22 Figure 1.2 The Mitochondrial Electron Transport Chain (ETC) + Complex V (ATP Synthase).................................................................................................26 Figure 1.3 Fatty acid oxidation pathway and the ETF/ETF-QO System............27 Figure 1.4 The Krebs cycle ...............................................................................29 Figure 1.5 Complex III/ Q cycle.........................................................................32 Figure 1.6 The Structure of Complex V (ATP Synthase) ..................................35 Figure 1.7 Redox States of CoQ10 Ubiquinone, Ubisemiquinone and Ubiquinol: the fully oxidized ubiquinone form (CoQ10); the radical semiquinone intermediate (CoQ10H·), and the fully reduced ubiquinol form (CoQ10H2). ........39 Figure 1.8 The Mevalonate Pathway-illustrating the synthesis of CoQ10 ..........46 Figure 1.9 CoQ Biosynthesis ............................................................................54 Figure 2.1 Haemocytometer .............................................................................65 Figure 2.2 SH-SY5Y Neuroblastoma Cells stained with Bodipy 581/591, a Lipid Peroxidation Marker..........................................................................................68 Figure 2.3 A typical Bovine Serum Albumin (BSA) standard line......................69 Figure 2.4 A Typical CoQ10 Chromatogram for a SH-SY5Y Cell Sample..........72 Figure 2.5 Concentration Curve for CoQ10 (HPLC-UV Detection Method)........72 Figure 2.6 Schematic Diagram of a High Performance Liquid Chromatography (HPLC) System.................................................................................................73 Figure 2.7 A Typical Reduced glutathione (GSH) Chromatogram for a SH-SY5Y Cell Sample.......................................................................................................75 Figure 2.8 Reduced Glutathione (GSH) A) Voltamagram- utilised to find the optimum electrochemical detector (EC) electrode voltage.
Recommended publications
  • Supplementary Materials: Evaluation of Cytotoxicity and Α-Glucosidase Inhibitory Activity of Amide and Polyamino-Derivatives of Lupane Triterpenoids

    Supplementary Materials: Evaluation of Cytotoxicity and Α-Glucosidase Inhibitory Activity of Amide and Polyamino-Derivatives of Lupane Triterpenoids

    Supplementary Materials: Evaluation of cytotoxicity and α-glucosidase inhibitory activity of amide and polyamino-derivatives of lupane triterpenoids Oxana B. Kazakova1*, Gul'nara V. Giniyatullina1, Akhat G. Mustafin1, Denis A. Babkov2, Elena V. Sokolova2, Alexander A. Spasov2* 1Ufa Institute of Chemistry of the Ufa Federal Research Centre of the Russian Academy of Sciences, 71, pr. Oktyabrya, 450054 Ufa, Russian Federation 2Scientific Center for Innovative Drugs, Volgograd State Medical University, Novorossiyskaya st. 39, Volgograd 400087, Russian Federation Correspondence Prof. Dr. Oxana B. Kazakova Ufa Institute of Chemistry of the Ufa Federal Research Centre of the Russian Academy of Sciences 71 Prospeсt Oktyabrya Ufa, 450054 Russian Federation E-mail: [email protected] Prof. Dr. Alexander A. Spasov Scientific Center for Innovative Drugs of the Volgograd State Medical University 39 Novorossiyskaya st. Volgograd, 400087 Russian Federation E-mail: [email protected] Figure S1. 1H and 13C of compound 2. H NH N H O H O H 2 2 Figure S2. 1H and 13C of compound 4. NH2 O H O H CH3 O O H H3C O H 4 3 Figure S3. Anticancer screening data of compound 2 at single dose assay 4 Figure S4. Anticancer screening data of compound 7 at single dose assay 5 Figure S5. Anticancer screening data of compound 8 at single dose assay 6 Figure S6. Anticancer screening data of compound 9 at single dose assay 7 Figure S7. Anticancer screening data of compound 12 at single dose assay 8 Figure S8. Anticancer screening data of compound 13 at single dose assay 9 Figure S9. Anticancer screening data of compound 14 at single dose assay 10 Figure S10.
  • Establishing the Pathogenicity of Novel Mitochondrial DNA Sequence Variations: a Cell and Molecular Biology Approach

    Establishing the Pathogenicity of Novel Mitochondrial DNA Sequence Variations: a Cell and Molecular Biology Approach

    Mafalda Rita Avó Bacalhau Establishing the Pathogenicity of Novel Mitochondrial DNA Sequence Variations: a Cell and Molecular Biology Approach Tese de doutoramento do Programa de Doutoramento em Ciências da Saúde, ramo de Ciências Biomédicas, orientada pela Professora Doutora Maria Manuela Monteiro Grazina e co-orientada pelo Professor Doutor Henrique Manuel Paixão dos Santos Girão e pela Professora Doutora Lee-Jun C. Wong e apresentada à Faculdade de Medicina da Universidade de Coimbra Julho 2017 Faculty of Medicine Establishing the pathogenicity of novel mitochondrial DNA sequence variations: a cell and molecular biology approach Mafalda Rita Avó Bacalhau Tese de doutoramento do programa em Ciências da Saúde, ramo de Ciências Biomédicas, realizada sob a orientação científica da Professora Doutora Maria Manuela Monteiro Grazina; e co-orientação do Professor Doutor Henrique Manuel Paixão dos Santos Girão e da Professora Doutora Lee-Jun C. Wong, apresentada à Faculdade de Medicina da Universidade de Coimbra. Julho, 2017 Copyright© Mafalda Bacalhau e Manuela Grazina, 2017 Esta cópia da tese é fornecida na condição de que quem a consulta reconhece que os direitos de autor são pertença do autor da tese e do orientador científico e que nenhuma citação ou informação obtida a partir dela pode ser publicada sem a referência apropriada e autorização. This copy of the thesis has been supplied on the condition that anyone who consults it recognizes that its copyright belongs to its author and scientific supervisor and that no quotation from the
  • Protective Effects of Alpha-Lipoic Acid and Coenzyme Q10 on Lipopolysaccharide-Induced Liver Injury in Rats

    Protective Effects of Alpha-Lipoic Acid and Coenzyme Q10 on Lipopolysaccharide-Induced Liver Injury in Rats

    Available online a t www.scholarsresearchlibrary.com Scholars Research Library Der Pharmacia Lettre, 2016, 8 (19):176-182 (http://scholarsresearchlibrary.com/archive.html) ISSN 0975-5071 USA CODEN: DPLEB4 Protective effects of alpha-lipoic acid and coenzyme Q10 on lipopolysaccharide-induced liver injury in rats Amr M. Emam 1, Gehan S. Georgy 2, Olfat G. Shaker 3, Hala M. Fawzy 2 and Hala F. Zaki 4 1Department of Pharmacology and Toxicology, Faculty of Pharmacy, Misr University for Science and Technology, 6th of October, El motamaiz, Cairo, Egypt 2Department of Pharmacology, National Organization for Drug Control and Research (NODCAR), Giza, Egypt 3Department of Biochemistry, Faculty of Medicine, Cairo University, Kasr El-Aini Street, Cairo, Egypt 4Department of Pharmacology and Toxicology, Faculty of Pharmacy, Cairo University, Kasr El-Aini Street, Cairo, Egypt _____________________________________________________________________________________________ ABSTRACT Lipopolysaccharide (LPS) is a major cell wall component of gram-negative bacteria known to stimulate the synthesis and secretion of several toxic metabolites, such as reactive oxygen species and cytokines. In this study, the protective effect of alpha-lipoic acid (ALA) and coenzyme Q10 (CoQ10) were evaluated in LPS-induced hepatic injury in rats. To this end, male adult Sprague Dawley rats were divided into five groups; normal control, LPS control where rats were injected with an initial dose of LPS (4 mg/kg; i.p.) on the 1 st day of the experiment followed by a challenging dose (2 mg/kg; i.p.) on the 8 th day, ALA (50 mg/kg), CoQ10 (10 mg/kg) and ALA plus CoQ10. Treatments continued for 15 days and the last three groups also received LPS.
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus

    A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus

    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
  • Energy + Coq10, NADH & B12

    Energy + Coq10, NADH & B12

    Energy + CoQ10, NADH & B12 it is mainly biosynthesized and concentrated in tissues with high energy turnover. Loss of function in mitochondria, the key organelle responsible for cellular energy production, can result in excess fatigue and other symptoms that are common complaints in almost every chronic disease. At the molecular level, a reduction in mitochondrial function occurs because of the following changes: (1) a loss of maintenance of the electrical and chemical transmembrane potential of the inner mitochondrial membrane, (2) alterations in the function of the electron transport chain, or (3) a reduction in the transport of critical metabolites into mitochondria. In turn, these changes result in reduced efficiency of oxidative phosphorylation and a reduction in adenosine-5’-triphosphate (ATP) production. Several components of this system require routine replacement, which can be facilitated with natural supplements. Clinical trials have shown the utility of using oral replacement supplements, such as coenzyme Q10 (CoQ10 [ubiquinone]), reduced nicotinamide adenine dinucleotide (NADH) and other supplements. Combinations of these supplements can significantly reduce fatigue and other symptoms associated with chronic disease and can naturally restore mitochondrial function, even in long-term patients with intractable fatigue.1 NADH is vital for many human body processes including muscle energy production. Skeletal muscle enables posture, breathing, and locomotion. Skeletal muscle also impacts systemic processes such as metabolism, thermoregulation, and immunity. Skeletal muscle is energetically expensive and is a major consumer of glucose and fatty acids. Metabolism INGREDIENTS of fatty acids and glucose requires NADH, which functions as a hydrogen/ Coenzyme Q10 (CoQ10) electron transfer molecule. Therefore, NADH plays a vital role in energy Coenzyme Q10 is a vitamin-like nutrient central to energy production at production.
  • Risk Assessment for Coenzyme Q10 (Ubiquinone) ଝ

    Risk Assessment for Coenzyme Q10 (Ubiquinone) ଝ

    Regulatory Toxicology and Pharmacology 45 (2006) 282–288 www.elsevier.com/locate/yrtph Risk assessment for coenzyme Q10 (Ubiquinone) ଝ John N. Hathcock ¤, Andrew Shao Council for Responsible Nutrition, 1828 L Street, NW, Suite 900, Washington, DC 20036-5114, USA Received 24 March 2006 Abstract Coenzyme Q10 (CoQ10) widely occurs in organisms and tissues, and is produced and used as both a drug and dietary supplement. Increasing evidence of health beneWts of orally administered CoQ10 are leading to daily consumption in larger amounts, and this increase justiWes research and risk assessment to evaluate the safety. A large number of clinical trials have been conducted using a range of CoQ10 doses. Reports of nausea and other adverse gastrointestinal eVects of CoQ10 cannot be causally related to the active ingredient because there is no dose–response relationship: the adverse eVects are no more common at daily intakes of 1200 mg than at a 60 mg. Systematic evaluation of the research designs and data do not provide a basis for risk assessment and the usual safe upper level of intake (UL) derived from it unless the newer methods described as the observed safe level (OSL) or highest observed intake (HOI) are utilized. The OSL risk assessment method indicates that the evidence of safety is strong at intakes up to 1200 mg/day, and this level is identiWed as the OSL. Much higher levels have been tested without adverse eVects and may be safe, but the data for intakes above 1200 mg/day are not suYcient for a conWdent conclusion of safety. © 2006 Elsevier Inc.
  • Nourishing and Health Benefits of Coenzyme Q10 – a Review

    Nourishing and Health Benefits of Coenzyme Q10 – a Review

    Czech J. Food Sci. Vol. 26, No. 4: 229–241 Nourishing and Health Benefits of Coenzyme Q10 – a Review Martina BOREKOVÁ1, Jarmila HOJEROVÁ1, Vasiľ KOPRDA1 and Katarína BAUEROVÁ2 1Institute of Biotechnology and Food Science, Faculty of Chemical and Food Technology, Slovak University of Technology, Bratislava, Slovak Republic; 2Institute of Experimental Pharmacology, Slovak Academy of Sciences, Bratislava, Slovak Republic Abstract Boreková M., Hojerová J., Koprda V., Bauerová K. (2008): Nourishing and health benefits of coen- zyme Q10 – a review. Czech J. Food Sci., 26: 229–241. Coenzyme Q10 is an important mitochondrial redox component and endogenously produced lipid-soluble antioxidant of the human organism. It plays a crucial role in the generation of cellular energy, enhances the immune system, and acts as a free radical scavenger. Ageing, poor eating habits, stress, and infection – they all affect the organism’s ability to provide adequate amounts of CoQ10. After the age of about 35, the organism begins to lose the ability to synthesise CoQ10 from food and its deficiency develops. Many researches suggest that using CoQ10 supplements alone or in com- bination with other nutritional supplements may help maintain health of elderly people or treat some of the health problems or diseases. Due to these functions, CoQ10 finds its application in different commercial branches such as food, cosmetic, or pharmaceutical industries. This review article gives a survey of the history, chemical and physical properties, biochemistry and antioxidant activity of CoQ10 in the human organism. It discusses levels of CoQ10 in the organisms of healthy people, stressed people, and patients with various diseases. This paper shows the distribution and contents of two ubiquinones in foods, especially in several kinds of grapes, the benefits of CoQ10 as nutritional and topical supplements and its therapeutic applications in various diseases.
  • Magnesium Ascorbyl Phosphate and Coenzyme Q10 Protect Keratinocytes Against UVA Irradiation by Suppressing Glutathione Depletion

    Magnesium Ascorbyl Phosphate and Coenzyme Q10 Protect Keratinocytes Against UVA Irradiation by Suppressing Glutathione Depletion

    MOLECULAR MEDICINE REPORTS 6: 375-378, 2012 Magnesium ascorbyl phosphate and coenzyme Q10 protect keratinocytes against UVA irradiation by suppressing glutathione depletion TSANN-LONG HWANG1,2, CHIN-JU TSAI3, JHIH-LONG CHEN3, TZU-TSUNG CHANGCHIEN3, CHEE-CHAN WANG3 and CHI-MING WU3 1Department of Surgery, Chang Gung Memorial Hospital, Tao-Yuan; 2Department of Surgery, School of Medicine, Chang Gung University, Tao-Yuan; 3Department of Cosmetic Science, Vanung University, Tao-Yuan, Taiwan, R.O.C. Received December 17, 2011; Accepted March 14, 2012 DOI: 10.3892/mmr.2012.933 Abstract. The aim of this study was to investigate whether Introduction magnesium ascorbyl phosphate (MAP) and coenzyme Q10 (CoQ10) can protect keratinocytes against ultraviolet (UV)A Ultraviolet (UV) irradiation (200-400 nm) causes a number of irradiation by increasing the levels of glutathione (GSH). The acute and chronic skin effects, which can result in inflammation, cell survival fraction was 89.9% when the keratinocytes were immunosuppression, premature skin aging and the develop- irradiated with UVA at a dose of 4 J/cm2. The cell survival frac- ment of skin malignancies (1). UVA irradiation (320-400 nm), tions were 48.4, 9.1 and 4.8%, at doses of 8, 16 and 32 J/cm2, which is not absorbed in the ozone layer, comprises more than respectively. MAP was added to the cells prior to UVA irradia- 95% of the UV light that reaches the earth. UVA penetrates tion at a dose of 8 J/cm2 and then the cell viability was assayed. the epidermis and affects the epidermal and dermal layers of The cell survival fractions were 51.6, 55.5, 64.8 and 76.7%, when the skin.
  • Electron Transport Discovery Four Complexes Complex I: Nadhà Coqh2

    Electron Transport Discovery Four Complexes Complex I: Nadhà Coqh2

    BI/CH 422/622 OUTLINE: Pyruvate pyruvate dehydrogenase Krebs’ Cycle How did he figure it out? Overview 8 Steps Citrate Synthase Aconitase Isocitrate dehydrogenase Ketoglutarate dehydrogenase Succinyl-CoA synthetase Succinate dehydrogenase Fumarase Malate dehydrogenase Energetics Regulation Summary Oxidative Phosphorylation Energetics (–0.16 V needed for making ATP) Mitochondria Transport (2.4 kcal/mol needed to transport H+ out) Electron transport Discovery Four Complexes Complex I: NADHà CoQH2 Complex II: Succinateà CoQH2 2+ Complex III: CoQH2à Cytochrome C (Fe ) 2+ Complex IV: Cytochrome C (Fe ) à H2O Electron Transport à O2 Inhibitors of Electron Transport Big Drop! • Inhibitors all stop ET and ATP synthesis: very toxic! Spectral work Big Drop! NADH Cyto-a3 Cyto-c1 Big Drop! Cyto-b Cyto-c Cyto-a Fully reduced Flavin Cyto-c + rotenone + antimycin A 300 350 400 450 500 550 600 650 700 1 Electron Transport Electron-Transport Chain Complexes Contain a Series of Electron Carriers • Better techniques for isolating and handling mitochondria, and isolated various fractions of the inner mitochondrial membrane • Measure E°’ • They corresponded to these large drops, and they contained the redox compounds isolated previously. • When assayed for what reactions they could perform, they could perform certain redox reactions and not others. • When isolated, including isolating the individual redox compounds, and measuring the E°’ for each, it was clear that an electron chain was occurring; like a wire! • Lastly, when certain inhibitors were added, some of the redox reactions could be inhibited and others not. Site of the inhibition could be mapped. Electron Transport Electron-Transport Chain Complexes Contain a Series of Electron Carriers • Better techniques for isolating and handling mitochondria, and isolated various fractions of the inner mitochondrial membrane • Measure E°’ • They corresponded to these large drops, and they contained the redox compounds isolated previously.
  • Molecular Diagnostic Requisition

    Molecular Diagnostic Requisition

    BAYLOR MIRACA GENETICS LABORATORIES SHIP TO: Baylor Miraca Genetics Laboratories 2450 Holcombe, Grand Blvd. -Receiving Dock PHONE: 800-411-GENE | FAX: 713-798-2787 | www.bmgl.com Houston, TX 77021-2024 Phone: 713-798-6555 MOLECULAR DIAGNOSTIC REQUISITION PATIENT INFORMATION SAMPLE INFORMATION NAME: DATE OF COLLECTION: / / LAST NAME FIRST NAME MI MM DD YY HOSPITAL#: ACCESSION#: DATE OF BIRTH: / / GENDER (Please select one): FEMALE MALE MM DD YY SAMPLE TYPE (Please select one): ETHNIC BACKGROUND (Select all that apply): UNKNOWN BLOOD AFRICAN AMERICAN CORD BLOOD ASIAN SKELETAL MUSCLE ASHKENAZIC JEWISH MUSCLE EUROPEAN CAUCASIAN -OR- DNA (Specify Source): HISPANIC NATIVE AMERICAN INDIAN PLACE PATIENT STICKER HERE OTHER JEWISH OTHER (Specify): OTHER (Please specify): REPORTING INFORMATION ADDITIONAL PROFESSIONAL REPORT RECIPIENTS PHYSICIAN: NAME: INSTITUTION: PHONE: FAX: PHONE: FAX: NAME: EMAIL (INTERNATIONAL CLIENT REQUIREMENT): PHONE: FAX: INDICATION FOR STUDY SYMPTOMATIC (Summarize below.): *FAMILIAL MUTATION/VARIANT ANALYSIS: COMPLETE ALL FIELDS BELOW AND ATTACH THE PROBAND'S REPORT. GENE NAME: ASYMPTOMATIC/POSITIVE FAMILY HISTORY: (ATTACH FAMILY HISTORY) MUTATION/UNCLASSIFIED VARIANT: RELATIONSHIP TO PROBAND: THIS INDIVIDUAL IS CURRENTLY: SYMPTOMATIC ASYMPTOMATIC *If family mutation is known, complete the FAMILIAL MUTATION/ VARIANT ANALYSIS section. NAME OF PROBAND: ASYMPTOMATIC/POPULATION SCREENING RELATIONSHIP TO PROBAND: OTHER (Specify clinical findings below): BMGL LAB#: A COPY OF ORIGINAL RESULTS ATTACHED IF PROBAND TESTING WAS PERFORMED AT ANOTHER LAB, CALL TO DISCUSS PRIOR TO SENDING SAMPLE. A POSITIVE CONTROL MAY BE REQUIRED IN SOME CASES. REQUIRED: NEW YORK STATE PHYSICIAN SIGNATURE OF CONSENT I certify that the patient specified above and/or their legal guardian has been informed of the benefits, risks, and limitations of the laboratory test(s) requested.
  • Table S2.Up Or Down Regulated Genes in Tcof1 Knockdown Neuroblastoma N1E-115 Cells Involved in Differentbiological Process Anal

    Table S2.Up Or Down Regulated Genes in Tcof1 Knockdown Neuroblastoma N1E-115 Cells Involved in Differentbiological Process Anal

    Table S2.Up or down regulated genes in Tcof1 knockdown neuroblastoma N1E-115 cells involved in differentbiological process analysed by DAVID database Pop Pop Fold Term PValue Genes Bonferroni Benjamini FDR Hits Total Enrichment GO:0044257~cellular protein catabolic 2.77E-10 MKRN1, PPP2R5C, VPRBP, MYLIP, CDC16, ERLEC1, MKRN2, CUL3, 537 13588 1.944851 8.64E-07 8.64E-07 5.02E-07 process ISG15, ATG7, PSENEN, LOC100046898, CDCA3, ANAPC1, ANAPC2, ANAPC5, SOCS3, ENC1, SOCS4, ASB8, DCUN1D1, PSMA6, SIAH1A, TRIM32, RNF138, GM12396, RNF20, USP17L5, FBXO11, RAD23B, NEDD8, UBE2V2, RFFL, CDC GO:0051603~proteolysis involved in 4.52E-10 MKRN1, PPP2R5C, VPRBP, MYLIP, CDC16, ERLEC1, MKRN2, CUL3, 534 13588 1.93519 1.41E-06 7.04E-07 8.18E-07 cellular protein catabolic process ISG15, ATG7, PSENEN, LOC100046898, CDCA3, ANAPC1, ANAPC2, ANAPC5, SOCS3, ENC1, SOCS4, ASB8, DCUN1D1, PSMA6, SIAH1A, TRIM32, RNF138, GM12396, RNF20, USP17L5, FBXO11, RAD23B, NEDD8, UBE2V2, RFFL, CDC GO:0044265~cellular macromolecule 6.09E-10 MKRN1, PPP2R5C, VPRBP, MYLIP, CDC16, ERLEC1, MKRN2, CUL3, 609 13588 1.859332 1.90E-06 6.32E-07 1.10E-06 catabolic process ISG15, RBM8A, ATG7, LOC100046898, PSENEN, CDCA3, ANAPC1, ANAPC2, ANAPC5, SOCS3, ENC1, SOCS4, ASB8, DCUN1D1, PSMA6, SIAH1A, TRIM32, RNF138, GM12396, RNF20, XRN2, USP17L5, FBXO11, RAD23B, UBE2V2, NED GO:0030163~protein catabolic process 1.81E-09 MKRN1, PPP2R5C, VPRBP, MYLIP, CDC16, ERLEC1, MKRN2, CUL3, 556 13588 1.87839 5.64E-06 1.41E-06 3.27E-06 ISG15, ATG7, PSENEN, LOC100046898, CDCA3, ANAPC1, ANAPC2, ANAPC5, SOCS3, ENC1, SOCS4,
  • Electronic Supplementary Material (ESI) for Metallomics

    Electronic Supplementary Material (ESI) for Metallomics

    Electronic Supplementary Material (ESI) for Metallomics. This journal is © The Royal Society of Chemistry 2018 Uniprot Entry name Gene names Protein names Predicted Pattern Number of Iron role EC number Subcellular Membrane Involvement in disease Gene ontology (biological process) Id iron ions location associated 1 P46952 3HAO_HUMAN HAAO 3-hydroxyanthranilate 3,4- H47-E53-H91 1 Fe cation Catalytic 1.13.11.6 Cytoplasm No NAD biosynthetic process [GO:0009435]; neuron cellular homeostasis dioxygenase (EC 1.13.11.6) (3- [GO:0070050]; quinolinate biosynthetic process [GO:0019805]; response to hydroxyanthranilate oxygenase) cadmium ion [GO:0046686]; response to zinc ion [GO:0010043]; tryptophan (3-HAO) (3-hydroxyanthranilic catabolic process [GO:0006569] acid dioxygenase) (HAD) 2 O00767 ACOD_HUMAN SCD Acyl-CoA desaturase (EC H120-H125-H157-H161; 2 Fe cations Catalytic 1.14.19.1 Endoplasmic Yes long-chain fatty-acyl-CoA biosynthetic process [GO:0035338]; unsaturated fatty 1.14.19.1) (Delta(9)-desaturase) H160-H269-H298-H302 reticulum acid biosynthetic process [GO:0006636] (Delta-9 desaturase) (Fatty acid desaturase) (Stearoyl-CoA desaturase) (hSCD1) 3 Q6ZNF0 ACP7_HUMAN ACP7 PAPL PAPL1 Acid phosphatase type 7 (EC D141-D170-Y173-H335 1 Fe cation Catalytic 3.1.3.2 Extracellular No 3.1.3.2) (Purple acid space phosphatase long form) 4 Q96SZ5 AEDO_HUMAN ADO C10orf22 2-aminoethanethiol dioxygenase H112-H114-H193 1 Fe cation Catalytic 1.13.11.19 Unknown No oxidation-reduction process [GO:0055114]; sulfur amino acid catabolic process (EC 1.13.11.19) (Cysteamine