DOCSLIB.ORG

Peroxisomes and Central Nervous System Dysgenesis and Dysfunction

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Peroxisomes and Central Nervous System Dysgenesis and Dysfunction Developmental Neurobiulogy, edited by Philippe Evrard and Alexandra Minkowski. Nestle Nutrition Workshop Series, Vol. 12. Nestec Ltd.. Vevey/Raven Press, Ltd., New York © 1989. Peroxisomes and Central Nervous System Dysgenesis and Dysfunction Sidney L. Goldfischer Department of Pathology, Albert Einstein College of Medicine, The Bronx, New York 10461 Peroxisomes play a key role in a number of genetic diseases. These include disor- ders in which the activity of a peroxisomal enzyme is deficient and an extraordinary group of diseases in which the formation of the organelle itself is defective (1-3) (Table 1). DISORDERS OF PEROXISOMAL BIOGENESIS Zellweger's cerebrohepatorenal syndrome (CHRS) is the first disease in which defective formation of peroxisomes was described (4,5). Infants affected with this rare autosomal recessive disorder usually die within one year. Clinical features of the syndrome include a typical facial appearance, with hypertelorism, a high fore- head, and pursed lips; minor skeletal abnormalities; renal cortical cysts; and severe hepatic fibrosis. Iron storage is frequently seen in the early stage of the disease. The most prominent findings are in the central nervous system and include profound hy- potonia. Cerebral abnormalities include polymicrogyria, pachygyria, olivary dys- plasia, and defective neuronal migration. Gliosis and accumulations of lipid in glia are associated with myelin breakdown, and the disease has been described as a suda- nophilic leukodystrophy (6-9). Hepatocellular peroxisomes have not been detected in children with this disease. This remarkable finding has been confirmed by many laboratories (10-13). It is par- ticularly surprising in view of the fact that there are approximately 1,000 peroxi- somes in a normal human hepatocyte (14). In hepatocytes and proximal renal tubules, where peroxisomes also appear to be absent, their diameter is approxi- mately 0.5 microns and normally they are readily detectable. In addition to the per- oxisomal defects, structural and chemical abnormalities in mitochondria have been described (4,5,10,13). The most common has been the report of diminished oxida- tion of succinate; further on in the respiratory pathway, electron transport is normal. 63 64 PEROXISOMES AND CNS DYSGENESIS TABLE 1. Peroxisomal diseases Defective peroxisomal biogenesis syndromes Zellweger's cerebrohepatorenal syndrome neonatal adrenoleukodystrophy infantile Refsum's disease Deficiency of a peroxisomal enzymatic activity X-linked childhood adrenoleukodystrophy pseudo-Zellweger's syndrome (3-oxoacyl-coA thiolase deficiency) acatalasia cerebrotendinous xanthomatosis (?) A variety of chemical abnormalities are seen in patients with CHRS, and many of these reflect the peroxisomal deficiency. These include markedly increased concen- trations of pipecolic acid (15), very long chain fatty acids (VLCFA) in blood and tis- sues (16,17), and the presence of abnormal bile acid intermediates (18,19). The activity of a key peroxisomal membrane enzyme involved in the synthesis of plas- malogens, dihydroxyacetone phosphate acyltransferase (DHAP-AT), is reduced by approximately 90%, and tissue concentrations of plasmalogens are less than 10% of normal in cells and tissues from patients with CHRS (20-22). Assay of DHAP-AT has been utilized in the antenatal diagnosis of CHRS (23). Neonatal adrenoleukodystrophy (NALD) is another disease in which the forma- tion of hepatocellular peroxisomes is defective (24-26). Although not as severely affected as children with CHRS (affected individuals have survived for as long as 6 years), NALD patients also suffer from severe hypotonia and seizures. Plasma con- centrations of VLCFA are increased six- to ninefold and cultured fibroblasts have a markedly diminished capacity to oxidize these fatty acids. Tissue deposits of VLCFA are widespread. We have examined the hepatocytes in two cases of NALD, and in both instances hepatocellular peroxisomes are markedly reduced in size and number (27). They are less than 0.2 |xm in diameter, and one-tenth as numerous as in the normal hepatocyte. Indeed, they are so small and sparse that several reports have been published stating that they are absent in this disease (28). They can be identified unequivocally by incubation in a cytochemical medium that makes visible a marker peroxisomal enzyme, catalase. Other biochemical findings in this disease are similar to CHRS (27). These include elevated concentrations of pipecolic acid, the presence of abnormal bile acid intermediates, and reduced levels of DHAP-AT, a key enzyme in plasmalogen synthesis (Janna Collins personal communication, 1985). The infantile variant of Refsum's disease is the third disorder in which formation of peroxisomes appears to be defective (29). Evidence for this comes from cyto- chemical and ultrastructural studies of the liver (30) and from biochemical studies of serum and fibroblasts (31,32). The classical form of Refsum's syndrome, which is an autosomal recessive disorder, manifests itself in the first or second decade of life PEROXISOMES AND CNS DYSGENESIS 65 and is characterized by retinitis pigmentosa, peripheral polyneuropathy, and cerebel- lar ataxia. Affected individuals have tissue stores of phytanic acid, a 20-carbon fatty acid derived from plant material. Because the stored material is exogenous in origin, a restricted diet is an effective therapy in this condition. The defect has been attrib- uted to the failure in the first step of catabolism of phytanic acid, that is, the alpha oxidation of the terminal carboxyl group. Recently several cases have been de- scribed of young children with phytanic acid storage disease who manifested symp- toms from birth. Electron microscopic studies by Dr. Frank Roels (30) have demonstrated that hepatocytes do not contain recognizable peroxisomes in this con- dition, and biochemical studies (31,32) have shown increased concentrations of the bile acid intermediate trihydroxycoprostanoic acid (THCA), as well as VLCFA. We are not aware of any studies in which the role of peroxisomes in the oxidation of phytanic acid has been analyzed. However, deficient oxidation of phytanic acid has been described in fibroblasts of such patients. We have recently found that serum phytanic acid is increased in CHRS and NALD, two diseases in which peroxisomes are deficient, but not in diseases associated with a more specific enzymatic defect, such as X-linked adrenoleukodystrophy (ALD) or a multiple peroxisomal oxidative activity deficiency syndrome (pseudo-Zellweger's disease) (33,34). This suggests that phytanic acid accumulation is secondary to a deficiency of peroxisomes, and that peroxisomal oxidative activity plays a critical role in the catabolism of phytanic acid. It is noteworthy that the number of peroxisomes is not diminished in fibro- blasts from patients with the classical form of Refsum's disease (35). It is unclear at the present time whether the CHRS, NALD, and infantile form of Refsum's disease are distinct entities or represent varying degrees of expression of a single disorder. It has recently been demonstrated that the absence of peroxisomes in CHRS is not absolute (36). Fibroblasts from four patients with CHRS or related diseases were examined. Two were classical Zellweger patients who died at the age of 6 months, and two others were longer-lived atypical patients (5 years, and alive at 3 years) who probably had NALD. Peroxisomes were detected, but reduced in number in all of these cell lines. Morphometric analysis demonstrated a reduction of approximately 90% in the first group of classical CHRS patients, and of 60% in the patients with longer survival. It will require much more extensive study to deter- mine whether the severity of disease is a consistent reflection of the number of per- oxisomes. The apparent inconsistency between hepatocytes and fibroblasts may reflect variations in the degree to which the deficiency manifests itself in different tissues. Intestinal peroxisomes do not appear to be reduced in NALD (27). It is note- worthy that in mice with testicular feminization syndrome there is a marked reduc- tion in size and number of peroxisomes in the interstitial cells of the testes (37). It is evident that there is considerable overlap between these syndromes (16). Even apparently distinctive pathologic phenomena, such as normal adrenals in CHRS and adrenal atrophy in NALD, may be a reflection of the shorter life span of these individuals. Recent studies have shown functional adrenal insufficiency in children with Zellweger's CHRS (38). Cerebellar heterotopia and olivary dysplasia occur both in classical CHRS and in NALD, but these phenomena have not been de- 66 PEROXISOMES AND CNS DYSGENESIS scribed in the infantile form of Refsum's disease. Other neurological findings, such as retinitis pigmentosa and demyelination, have been observed in all of these condi- tions, even though the degree of involvement may vary considerably. Comparison of conditions in which peroxisomal biogenesis is defective with several other disor- ders in which there appears to be defective activity of a single peroxisomal enzyme may be instructive in relating specific pathologic events to specific enzymatic activi- ties. It must be stressed that there have been no studies of mitochondrial respiration in NALD and infantile Refsum's disease. PEROXISOMAL DISORDERS AFFECTING SPECIFIC ENZYMATIC ACTIVITIES A second group of peroxisomal disorders resemble the better known deficiency diseases. These are diseases in which the
Load more

Recommended publications
  • Newborn Screening for X-Linked Adrenoleukodystrophy: Information for Parents
    Newborn Screening for X-linked Adrenoleukodystrophy: Information for Parents Baby girl with ABCD1 gene mutation What is newborn screening? the symptoms of ALD during childhood. Rarely, some women who are carriers of ALD develop mild symptoms as adults. It Newborn screening involves laboratory testing on a small is important for your family to meet with a genetic counselor sample of blood collected from newborns’ heels. Every state to talk about the genetics of ALD and implications for other has a newborn screening program to identify infants with rare family members. disorders, which would not usually be detected at birth. Early diagnosis and treatment of these disorders often prevents Why do only boys have ALD? serious complications. Only boys have ALD because it is caused by a mutation in What is adrenoleukodystrophy (ALD)? a gene (ABCD1) on the X chromosome, called “X-linked inheritance.” Males only have one X chromosome so they have ALD is one of over 40 disorders included in newborn screening one ABCD1 gene. Males with a nonfunctioning ABCD1 gene in New York State. It is a rare genetic disorder. People with have ALD. Females have 2 X chromosomes, so they have two ALD are unable to breakdown a component of food called ABCD1 genes. Females with one ABCD1 gene mutation will very long chain fatty acids (VLCFA). If VLCFA are not broken be carriers. When a mother is a carrier of ALD, each son has a down, they build up in the body and cause symptoms. 50% chance of inheriting the disorder and each daughter has a 50% chance of being a carrier.
    [Show full text]
  • Antenatal Diagnosis of Inborn Errors Ofmetabolism
    816 ArchivesofDiseaseinChildhood 1991;66: 816-822 CURRENT PRACTICE Arch Dis Child: first published as 10.1136/adc.66.7_Spec_No.816 on 1 July 1991. Downloaded from Antenatal diagnosis of inborn errors of metabolism M A Cleary, J E Wraith The introduction of experimental treatment for Sample requirement and techniques used in lysosomal storage disorders and the increasing prenatal diagnosis understanding of the molecular defects behind By far the majority of antenatal diagnoses are many inborn errors have overshadowed the fact performed on samples obtained by either that for many affected families the best that can amniocentesis or chorion villus biopsy. For be offered is a rapid, accurate prenatal diag- some disorders, however, the defect is not nostic service. Many conditions remain at best detectable in this material and more invasive only partially treatable and as a consequence the methods have been applied to obtain a diagnos- majority of parents seek antenatal diagnosis in tic sample. subsequent pregnancies, particularly for those disorders resulting in a poor prognosis in terms of either life expectancy or normal neurological FETAL LIVER BIOPSY development. Fetal liver biopsy has been performed to The majority of inborn errors result from a diagnose ornithine carbamoyl transferase defi- specific enzyme deficiency, but in some the ciency and primary hyperoxaluria type 1. primary defect is in a transport system or Glucose-6-phosphatase deficiency (glycogen enzyme cofactor. In some conditions the storage disease type I) could also be detected by biochemical defect is limited to specific tissues this method. The technique, however, is inva- only and this serves to restrict the material avail- sive and can be performed by only a few highly able for antenatal diagnosis for these disorders.
    [Show full text]
  • Tests Performed Through Our International Collaboration
    Tests performed through our international collaboration Molecular Studies for Inborn Errors of Metabolism A Inborn Errors of Metabolism Defective Gene 1 Acyl - Co A oxidase deficiency ACOX 1 2 Argininosuccinate lyase deficiency ASL 3 Aromatic L – amino acid decarboxylase (AADC) DDC Deficiency 4 Carnitine – acylcarnitine translocase (CACT) CACT Deficiency 5 Primary (systemic) carnitine deficiency OCTN2 6 Carnitine palmitoyltransferase I (CPT1) CPT1A 7 Carnitine palmitoyltransferase 2 (CPT2) CPT2 8 CHILD Syndrome NSDHL 9 Conradi – Hunermann – Happle syndrome EBP (CDPX2) 10 D – Bifunctional protein (DBP) Deficiency DBP, MFE2 11 Desmosterolosis DHCR24 12 Dihydropyrimidinase (DHP) Deficiency DPYS 13 Dihydropyrimidine dehydrogenase (DPD) DPYD Deficiency 14 Ethylmalonaciduria ETHE1 (Ethylmalonic encephalopathy) 15 Fructose intolerance, hereditary ALDOB 16 Galactosemia, classic GALT 17 Galactokinase deficiency GALK1 18 Glutaric aciduria type I (Glutaryl – Co A GCDH dehydrogenase deficiency 19 Glycogen storage disease 0 GYS2 20 Greenberg skeletal dysplasia LBR (Sterol – delta 14 reductase deficiency) 21 GTP cyclohydrolase I deficiency GCH1 22 Hydroxyacyl – Co A dehydrogenase deficiency HADH2 (2 – Methyl – 3 – hydroxybutyryl – CoA dehydrogenase deficiency) 23 Hyper Ig D Syndrome MVK (Mevalonate Kinase deficiency) 24 Hyperoxaluria type I AGXT 25 Isovaleric acidemia IVD 26 Lathosterolosis SC5DL 27 3 – methylglutaconicaciduria type I (3 – AUH methylglutaconyl – CoA hydratase deficiency) 28 Medium – chain – acyl – Co A dehydrogenase ACADM (MCAD) Deficiency
    [Show full text]
  • Inherited Peroxisomal Disorders Involving the Nervous System
    Arch Dis Child: first published as 10.1136/adc.63.7.767 on 1 July 1988. Downloaded from Archives of Disease in Childhood, i988, 63, 767-770 Annotations Inherited peroxisomal disorders involving the nervous system For genetic reasons peroxisomes may be absent or defective. Because of the great importance of the lack one or more of their enzymes. The result may peroxisomal enzymes involved in the I oxidation of be subtle and long delayed, but is often catastrophic VLCFA and the use of measurements of VLCFA in and immediately recognisable as a serious disorder diagnosis, this group is itself subdivided. Group 3A at birth. The trouble for the clinician is that it is includes those conditions in which VLCFA oxidation often not possible to make a diagnosis unless the is impaired and thus VLCFA accumulates, and right tests are thought of: these tests are not part of group 3B those in which the single enzyme defect is routine 'metabolic screens.' It is first necessary to of another sort. frame the question 'could this be a peroxisomopathy?' The object of this annotation is to help with when GENERALISED PEROXISOMAL DISORDERS (GROUP 1) and how to answer this question. The now classical example is the cerebro-hepato- renal syndrome of Zellweger. l 2The typical neonate Peroxisomes is more dysmorphic than a baby with Down's syndrome with high forehead and huge fontanelle Peroxisomes are subcellular organelles with a single with metopic extension. Inactivity is even greater membrane which occur in every cell apart from the than in the Prader-Willi syndrome and hypotonia mature erythrocyte.
    [Show full text]
  • Thesis Final
    PEX1 Mutations in Australasian Patients with Disorders of Peroxisome Biogenesis Author Maxwell, Megan Amanda Published 2004 Thesis Type Thesis (PhD Doctorate) School School of Biomolecular and Biomedical Sciences DOI https://doi.org/10.25904/1912/1930 Copyright Statement The author owns the copyright in this thesis, unless stated otherwise. Downloaded from http://hdl.handle.net/10072/366184 Griffith Research Online https://research-repository.griffith.edu.au PEX1 MUTATIONS IN AUSTRALASIAN PATIENTS WITH DISORDERS OF PEROXISOME BIOGENESIS Megan Amanda Maxwell, BSc, MSc(Hons) School of Biomolecular and Biomedical Science, Faculty of Science, Griffith University Submitted in fulfilment of the requirements of the degree of Doctor of Philosophy July, 2003 ABSTRACT The peroxisome is a subcellular organelle that carries out a diverse range of metabolic functions, including the β-oxidation of very long chain fatty acids, the breakdown of peroxide and the α-oxidation of fatty acids. Disruption of peroxisome metabolic functions leads to severe disease in humans. These diseases can be broadly grouped into two categories: those in which a single enzyme is defective, and those known as the peroxisome biogenesis disorders (PBDs), which result from a generalised failure to import peroxisomal matrix proteins (and consequently result in disruption of multiple metabolic pathways). The PBDs result from mutations in PEX genes, which encode protein products called peroxins, required for the normal biogenesis of the peroxisome. PEX1 encodes an AAA ATPase that is essential for peroxisome biogenesis, and mutations in PEX1 are the most common cause of PBDs worldwide. This study focused on the identification of mutations in PEX1 in an Australasian cohort of PBD patients, and the impact of these mutations on PEX1 function.
    [Show full text]
  • Role of Catalase in Oxidative Stress-And Age-Associated
    Hindawi Oxidative Medicine and Cellular Longevity Volume 2019, Article ID 9613090, 19 pages https://doi.org/10.1155/2019/9613090 Review Article Role of Catalase in Oxidative Stress- and Age-Associated Degenerative Diseases Ankita Nandi,1 Liang-Jun Yan ,2 Chandan Kumar Jana,3 and Nilanjana Das 1 1Department of Biotechnology, Visva-Bharati University, Santiniketan, West Bengal 731235, India 2Department of Pharmaceutical Sciences, UNT System College of Pharmacy, University of North Texas Health Science Center, Fort Worth, TX 76107, USA 3Department of Chemistry, Purash-Kanpur Haridas Nandi Mahavidyalaya, P.O. Kanpur, Howrah, West Bengal 711410, India Correspondence should be addressed to Nilanjana Das; [email protected] Received 25 March 2019; Revised 18 June 2019; Accepted 14 August 2019; Published 11 November 2019 Academic Editor: Cinzia Signorini Copyright © 2019 Ankita Nandi et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Reactive species produced in the cell during normal cellular metabolism can chemically react with cellular biomolecules such as nucleic acids, proteins, and lipids, thereby causing their oxidative modifications leading to alterations in their compositions and potential damage to their cellular activities. Fortunately, cells have evolved several antioxidant defense mechanisms (as metabolites, vitamins, and enzymes) to neutralize or mitigate the harmful effect of reactive species and/or their byproducts. Any perturbation in the balance in the level of antioxidants and the reactive species results in a physiological condition called “oxidative stress.” A catalase is one of the crucial antioxidant enzymes that mitigates oxidative stress to a considerable extent by destroying cellular hydrogen peroxide to produce water and oxygen.
    [Show full text]
  • Pseudo Infantile Refsum's Disease: Catalase- Deficient Peroxisomal Particles with Partial Deficiency of Plasmalogen Synthesis and Oxidation of Fatty Acids
    003 I-3998/93/3403-0270$03.00/0 PEDIATRIC RESEARCH Vol. 34. No. 3. 1993 Copyright 8 1993 International Pediatric Research Foundation. Inc I'ritrrid in U S :I. Pseudo Infantile Refsum's Disease: Catalase- Deficient Peroxisomal Particles with Partial Deficiency of Plasmalogen Synthesis and Oxidation of Fatty Acids P. AUBOURG, K. KREMSER, M. 0. ROLAND. F. ROCCHICCIOLI. AND I. SlNGH ABSTRACT. Zellweger syndrome, neonatal adrenoleuko- RCDP, rhizomelic chondrodysplasia punctata dystrophy, and infantile Refsum's disease are genetic dis- IRD, infantile Refsum's disease orders characterized by the virtual absence of catalase- positive perosisomes and a general impairment of perosi- somal functions. Recent studies in these three disorders have provided morphologic evidence of perosisomal Peroxisomes are subcellular organelles that participate in the "ghosts" of density 1.10 g/cd that contain membrane @-oxidation of long-chain fatty acids and VLCFA (1-3), the proteins but lack a majority of the matrix enzyme activities. synthesis of plasmalogens (4), the oxidation of L-pipecolic acid We report here the biochemical studies in a female infant (5, 6). and bile acid synthesis (7). In addition, peroxisomes with clinical features of infantile Refsum's disease whose contain catalase that degrades the Hz02 produced by various liver and fibroblasts contained cytosolic catalase but no oxidases (8). Peroxisomes, like mitochondria. arise by growth catalase-positive peroxisomes. Oxidation of phytanic and and division of preexisting peroxisomes (9). All peroxisomal pipecolic acids was severely impaired, whereas osidation proteins studied so far are synthesized on free polyribosomes, of very-long-chain fatty acids and dihydrosyacetone phos- then translocated into preexisting peroxisomes.
    [Show full text]
  • HSAPQ State Series - 2014 State Finals Round 2 First Period: Tossups with Bonus
    HSAPQ State Series - 2014 State Finals Round 2 First Period: Tossups with Bonus 1. A specialized type of this organelle found in plants is the site of the glyoxylate (glai-OCK-sul-ate) cycle. Acatalasia (UH-cat-uh-LAHZ-ee-uh) results when this organelle lacks a key enzyme. Most disorders involving this organelle, such as infantile Refsum disease, result from mutations in PEX genes. A sharp reduction in the number of this organelle is observed in patients with (*) Zellweger syndrome. This organelle contains the enzyme catalase (CAT-uh-layz). For 10 points, name this organelle that decomposes its namesake compound into oxygen and water. ANSWER: peroxisome 127-14-102-02101 BONUS: What Frenchman created an international incident in 1793 when he traveled through the U.S. trying to raise anti-British sentiment and arm privateers to fight with the French? ANSWER: Edmond-Charles Genet [or Citizen Genet] 015-14-102-0210-11 2. This mountain was first climbed by Edwin James, who described seeing the blue columbine during the climb. Its namesake was killed in the Battle of York during the War of 1812 and had previously explored the west on orders of James Wilkinson. Katharine Lee (*) Bates wrote the song "America the Beautiful" after admiring the view from this mountaintop. During the "Fifty-Niner Gold Rush," miners used a slogan advocating this mountain "or bust." For 10 points, name this peak in the Rocky Mountains, which is named for an explorer whose first name was Zebulon. ANSWER: Pike's Peak 052-14-102-02102 BONUS: What first-in-first-out data structure comes in a "priority" variety? ANSWER: queue 014-14-102-0210-11 3.
    [Show full text]
  • Diseases Catalogue
    Diseases catalogue AA Disorders of amino acid metabolism OMIM Group of disorders affecting genes that codify proteins involved in the catabolism of amino acids or in the functional maintenance of the different coenzymes. AA Alkaptonuria: homogentisate dioxygenase deficiency 203500 AA Phenylketonuria: phenylalanine hydroxylase (PAH) 261600 AA Defects of tetrahydrobiopterine (BH 4) metabolism: AA 6-Piruvoyl-tetrahydropterin synthase deficiency (PTS) 261640 AA Dihydropteridine reductase deficiency (DHPR) 261630 AA Pterin-carbinolamine dehydratase 126090 AA GTP cyclohydrolase I deficiency (GCH1) (autosomal recessive) 233910 AA GTP cyclohydrolase I deficiency (GCH1) (autosomal dominant): Segawa syndrome 600225 AA Sepiapterin reductase deficiency (SPR) 182125 AA Defects of sulfur amino acid metabolism: AA N(5,10)-methylene-tetrahydrofolate reductase deficiency (MTHFR) 236250 AA Homocystinuria due to cystathionine beta-synthase deficiency (CBS) 236200 AA Methionine adenosyltransferase deficiency 250850 AA Methionine synthase deficiency (MTR, cblG) 250940 AA Methionine synthase reductase deficiency; (MTRR, CblE) 236270 AA Sulfite oxidase deficiency 272300 AA Molybdenum cofactor deficiency: combined deficiency of sulfite oxidase and xanthine oxidase 252150 AA S-adenosylhomocysteine hydrolase deficiency 180960 AA Cystathioninuria 219500 AA Hyperhomocysteinemia 603174 AA Defects of gamma-glutathione cycle: glutathione synthetase deficiency (5-oxo-prolinuria) 266130 AA Defects of histidine metabolism: Histidinemia 235800 AA Defects of lysine and
    [Show full text]
  • Catalase ECAT005.Pdf
    BioAssay Systems Catalase ECAT005.pdf EnzyChrom TM Catalase Assay Kit (ECAT-100) Quantitative Colorimetric/Fluorimetric Catalase Determination DESCRIPTION CATALASE (EC 1.11.1.6), is an ubiquitous antioxidant enzyme that 95 µL Assay Buffer. Note: diluted H 2O2 is not stable. Prepare fresh catalyzes the decomposition of hydrogen peroxide (H2O2) to water and dilutions for each experiment. oxygen. Add 90 µL of the 50 µM Substrate to these wells to initiate the catalase catalase reaction. Tap plate quick to mix. Incubate 30 min at room temperature. During the incubation time, proceed with Steps 3 and 4 below. 2 H 2O2 O2 + 2 H 2O By preventing excessive H 2O2 build up, catalase allows important cellular 3. H2O2 Standard Curve . Mix 40µL of the 4.8 mM H2O2 with 440 µL processes which produce H 2O2 as a byproduct to occur safely. dH2O to yield 400 µM H2O2. Prepare standards as shown in the Table Deficiency in catalase activity has been associated with grey hair and below. Transfer 10 µL standards into separate wells of the 96-well peroxisomal disorder acatalasia. Simple, direct and high-throughput plate. Add 90 µL Assay Buffer to the standards. assays for catalase activity find wide applications. BioAssay Systems' improved assay directly measures catalase degradation of H 2O2 using a No 400 µM H2O2 + H 2O Vol ( µL) H2O2 ( µM) redox dye. The change in color intensity at 570nm or fluorescence 1 100 µL + 0 µL 100 400 intensity ( λem/ex = 585/530nm) is directly proportional to the catalase 2 60 µL + 40 µL 100 240 activity in the sample.
    [Show full text]
  • Lessons Learned
    Prevention and Reversal of Chronic Disease Copyright © 2019 RN Kostoff PREVENTION AND REVERSAL OF CHRONIC DISEASE: LESSONS LEARNED By Ronald N. Kostoff, Ph.D., School of Public Policy, Georgia Institute of Technology 13500 Tallyrand Way, Gainesville, VA, 20155 [email protected] KEYWORDS Chronic disease prevention; chronic disease reversal; chronic kidney disease; Alzheimer’s Disease; peripheral neuropathy; peripheral arterial disease; contributing factors; treatments; biomarkers; literature-based discovery; text mining ABSTRACT For a decade, our research group has been developing protocols to prevent and reverse chronic diseases. The present monograph outlines the lessons we have learned from both conducting the studies and identifying common patterns in the results. The main product of our studies is a five-step treatment protocol to reverse any chronic disease, based on the following systemic medical principle: at the present time, removal of cause is a necessary, but not necessarily sufficient, condition for restorative treatment to be effective. Implementation of the five-step treatment protocol is as follows: FIVE-STEP TREATMENT PROTOCOL TO REVERSE ANY CHRONIC DISEASE Step 1: Obtain a detailed medical and habit/exposure history from the patient. Step 2: Administer written and clinical performance and behavioral tests to assess the severity of symptoms and performance measures. Step 3: Administer laboratory tests (blood, urine, imaging, etc) Step 4: Eliminate ongoing contributing factors to the chronic disease Step 5: Implement treatments for the chronic disease This individually-tailored chronic disease treatment protocol can be implemented with the data available in the biomedical literature now. It is general and applicable to any chronic disease that has an associated substantial research literature (with the possible exceptions of individuals with strong genetic predispositions to the disease in question or who have suffered irreversible damage from the disease).
    [Show full text]
  • 1 a Clinical Approach to Inherited Metabolic Diseases
    1 A Clinical Approach to Inherited Metabolic Diseases Jean-Marie Saudubray, Isabelle Desguerre, Frédéric Sedel, Christiane Charpentier Introduction – 5 1.1 Classification of Inborn Errors of Metabolism – 5 1.1.1 Pathophysiology – 5 1.1.2 Clinical Presentation – 6 1.2 Acute Symptoms in the Neonatal Period and Early Infancy (<1 Year) – 6 1.2.1 Clinical Presentation – 6 1.2.2 Metabolic Derangements and Diagnostic Tests – 10 1.3 Later Onset Acute and Recurrent Attacks (Late Infancy and Beyond) – 11 1.3.1 Clinical Presentation – 11 1.3.2 Metabolic Derangements and Diagnostic Tests – 19 1.4 Chronic and Progressive General Symptoms/Signs – 24 1.4.1 Gastrointestinal Symptoms – 24 1.4.2 Muscle Symptoms – 26 1.4.3 Neurological Symptoms – 26 1.4.4 Specific Associated Neurological Abnormalities – 33 1.5 Specific Organ Symptoms – 39 1.5.1 Cardiology – 39 1.5.2 Dermatology – 39 1.5.3 Dysmorphism – 41 1.5.4 Endocrinology – 41 1.5.5 Gastroenterology – 42 1.5.6 Hematology – 42 1.5.7 Hepatology – 43 1.5.8 Immune System – 44 1.5.9 Myology – 44 1.5.10 Nephrology – 45 1.5.11 Neurology – 45 1.5.12 Ophthalmology – 45 1.5.13 Osteology – 46 1.5.14 Pneumology – 46 1.5.15 Psychiatry – 47 1.5.16 Rheumatology – 47 1.5.17 Stomatology – 47 1.5.18 Vascular Symptoms – 47 References – 47 5 1 1.1 · Classification of Inborn Errors of Metabolism 1.1 Classification of Inborn Errors Introduction of Metabolism Inborn errors of metabolism (IEM) are individually rare, but collectively numerous.
    [Show full text]