DOCSLIB.ORG

In Fibroblasts by Intracellular Concentrations of Cyclic Adenosine Monophosphate

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
In Fibroblasts by Intracellular Concentrations of Cyclic Adenosine Monophosphate Proc. Nat. Acad. Sci. USA Vol. 69, No. 2, pp. 459-462, February 1972 Regulation of Adenosine 3':5'-Cyclic Monophosphate Phosphodiesterase Activity in Fibroblasts by Intracellular Concentrations of Cyclic Adenosine Monophosphate (3T3/dibutyryl cyclic AMP/SV40-transformed cells/Michaelis constants/L cells/prostaglandin El) MASSIMO D'ARMIENTO, GEORGE S. JOHNSON, AND IRA PASTAN Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014 Communicated by Earl W. Sutherland, December 10, 1971 ABSTRACT Cyclic AMP-phosphodiesterase is present also obtained from Schwarz-Mann, snake venom (Ophiophagus in various mouse fibroblasts. Contact-inhibited 3T3 cells hannah) from Sigma Chemical Co., and Dowex 1-X2, 200-400 contain two forms of the enzyme, one with a Km of 2.5 uIM and the second with a Km of 71 AM. As 3T3 cells grow to mesh from Bio-Rad. The resin was washed with 0.5 N NaOH, confluency and cAMP concentrations rise, the activity of 0.5 N HCl, and deionized water, to a final pH of 5.0, before the first enzyme increases, whereas that of the second is use. unchanged. A line of SV40-transformed 3T3 cells with low cAMP concentrations also has low levels of the cAMP- Separatipn and Assay of phosphodiesterase. The cells were phosphodiesterase with a K,,, of 2.5 1AM. Treatment of 3T3 grown under standard conditions (4). The cAMP phosphodi- and SV40-transformed 3T3 cells with dibutyryl cAMP and theophylline increases cAMP-phosphodiesterase accumu- esterase was prepared by washing the monolayer four times lation. This accumulation is blocked by cycloheximide and with ice-cold phosphate-buffered saline (pH 7.4) and four actinomycin D. The newly formed enzyme resembles the times with ice-cold homogenizing medium containing 0.04 M higher affinity enzyme present in unstimulated cells, Tris- HCl (pH 8.0)-10.9% sucrose (w/v). The cells were since it has a Km of 1.2-2.0 MM, and is stimulated by snake scraped from the culture dish with a rubber stopper, and venom. In L cells in which cAMP concentrations are ele- vated by treatment with prostaglandin El, cAMP phos- homogenized in 0.5-1.0 ml of homogenizing medium, at a phodiesterase also accumulates. We conclude that intra- final concentration of protein of 2-5 mg/ml, in a glass Dounce cellular concentrations of cAMP regulate the synthesis of homogenizer (tight pestle; 20 strokes). All steps were at 0- cAMP-phosphodiesterase, and that cAMP functions as an 4°C. The resulting homogenate was used directly as the inducer of the enzyme. enzyme in the assay. Protein measurements were made by the Previous studies indicated that cAMP regulates the growth, method of Lowry et al. (6), with bovine serum albumin as morphology, and adhesiveness of normal and transformed the standard. The assay for cAMP phosphodiesterase was fibroblasts (1-4). Thus, an understanding of cAMP metabo- essentially the two-step assay of Butcher and Sutherland (7), lism is important to elucidate the regulation of these proper- with snake venom used in the second step as a source of ties in normal and cancer cells. 5'-nucleotidase to convert the product of cAMP phospho- cAMP concentrations are determined by the activity of the diesterase (i.e., 5'-AMP) to adenosine and inorganic phos- enzyme adenylate cyclase, which catalyzes the formation of phate. cAMP from ATP, and cAMP phosphodiesterase, which In the first stage, the 0.18-ml reaction mixture contained 40 degrades cAMP into 5'-AMP (for review, see ref. 5). cAMP mM Tris HCl (pH 8), 10 mM MgCl2, 4 mM 2-mercapto- can also leak from cells into the extracellular fluid. We have ethanol, and an appropriate concentration of enzyme. After an previously reported on the properties of adenylate cyclase in equilibration period of 10 min at 300C, during which no loss of normal and transformed fibroblasts (4). Here, we report on the enzyme activity occurred, the reaction was initiated by the properties of the cAMP phosphodiesterase. addition of 20 ul of a solution containing [3H]cAMP (200,000 We find that mouse 3T3 cells have high levels of cAMP cpm) and unlabeled cAMP. After 10 min at 300C, the reaction phosphodiesterase activity; kinetic analysis, suggests the was stopped by heating at 1000C for 1 min. 50 ug of snake activity is due to two enzymes. However, one line of SV4O- venom was added and the components were incubated for a transformed cells has very low activities of cAMP phosphodi- further 10 min at 30°C to convert 5'-AMP to adenosine. This esterase. The total activity of the enzyme is dependent on the reaction was stopped by the addition of 1.0 ml of a 1:3 slurry intracellular concentration of cAMP, and this activity is in- of Bio-Rad AG 1X-2 resin; after they were mixed, the tubes creased by treatment of cells with But2cAMP (N6-2'-O- were centrifuged 10 min at 4000 X g and the radioactivity in dibutyryl cAMP) and theophylline. These findings suggest the supernatant was measured. When snake venom was used that cAMP functions as an inducer of the enzyme. to stimulate the cAMP phosphodiesterase (8), 50 ,g was present in the first stage of the reaction. The blank for each MATERIALS AND METHODS experiment was prepared with heat-inactivated enzyme, or by [3H]cAMP (14.3 Ci/mmole) was obtained from Schwarz- omission of the enzyme from the incubation mixture. The Mann and purified on Dowex AG 50W-X8 resin. cAMP was blank value was subtracted from each experimental value. In all assays the rate of [3H]cAMP hydrolysis was linear for at Abbreviations: But2cAMP, N'-2'-O-dibutyryl cAMP. least 15 min and directly proportional to protein concentration 459 Downloaded by guest on September 27, 2021 460 Biochemistry: d'Armiento et al. Proc. Nat. Acad. Sci. USA 69 (1972) E- .E E c E 8 - B -..0 C .E A S.4 0O-, 4 ~ ~ ~ ~ ~ I-E -0~ ~~~~0 2 1 1.6 32 4 53 8 .026 .040 .064 .OeO .106 .160 I/cAWMPxIOI "M I/cAMPxI M FIG. 2. Kinetic analysis by double-reciprocal plot of cAMP FIG. 1A and B. Kinetic analysis by double-reciprocal plot phosphodiesterase activity in 3T3-SV40 cells. 0-- 0, cAMP of cAMP phosphodiesterase activity in 3T3-4 cells: --- -0, phosphodiesterase activity in the presence of venom: 0-0, cAMP phosphodiesterase activity in the presence of snake cAMP phosphodiesterase activity in the absence of venom. venom: 0-*, cAMP phosphodiesterase activity in the absence of venom. appear to contain two enzymatic activities. One has an ap- parent Km of 2.5,uM and a Vm.x of 200 pmol per mg of protein over a range of 5-250 j&g/ml. In each assay not more than 15- per min (Fig. 1A). The increase of Vm.. due to the presence of 20% of the substrate was consumed. The relative velocity was snake venom in the first step of the reaction is from 200 to 300 expressed in nmol of cAMP hydrolyzed per mg of protein per pmol per mg protein per min. The second enzymatic activity min at 30'C. All of the experiments were repeated from 3 to 5 has an apparent Km of 71 ,uM and a Vma. of 2500 pmol per mg times, and each experimental value represents the mean of two protein per min (Fig. 1B). The presence of snake venom seems or three closely agreeing numbers. to slightly decrease the activity of this second enzyme. RESULTS Table 1 shows the increase in the cAMP phosphodiesterase activity during growth of 3T34 cells. When these cells become Kinetic analysis of the cAMP phosphodiesterase activity in the cAMP phosphodiesterase activity rises Fig. 1A and contact-inhibited, 3T34 cells by double-reciprocal plots is shown in from to 5-fold. This rise is present in both the low- and the B. We find that these cells, like many tissues of the rat (9), 3- high-Km enzymes, and occurs whether the enzyme is measured in the presence or in the absence of venom. The apparent Km TABLE 1. cAMP phosphodiesterase actinty during the growth of 3TS cells TABLE 2. cAMP phosphodiesterase activity in STS-4 cells treated with But2cAMP and theophylline Log-phase Contact-inhibited growing cells cells cAMP phospho- Cyclic AMP (pM) diesterase activity (ref. 3) 3.3 11 (pmol/mg of protein Venom - + - + per min) Vma.. (nmol per Venom mg of protein _+ per min)* 0.1 0.2 0.2 0.3 V.., (nmol per Control 26 48 mg of protein Butc2AMP and theophylline 67 103 per mini)t 1.4 0.6 2.2 2.9 But2cAMP (1.2 mM) and theophylline (1 mM) were added * K., was 2,uM for all four conditions. for 40 hr to a 4-day-old culture. Enzyme activity was measured at t Kin was 71 uM for all four conditions. 0.625 uM cAMP. Downloaded by guest on September 27, 2021 Proc. Nat. Acad. Sci. USA 69 (1972) Cyclic AMP-Phosphodiesterase in Fibroblasts 461 values, however, seem to be unchanged. We found that when 3T3-4 cells become contact-inhibited, the intracellular con- centration of cAMP rises by about 3 to 4-fold (3). Therefore, it seems likely that cAMP itself is the inducer of the cAMP phosphodiesterase. To test this possibility, we treated 3T3-4 cells with But2- cAMP and theophylline. The results of these experiments are shown in Table 2. After 40 hr of treatment with 1.2 mM But2cAMP and 1mM theophylline, the cAMP phospho- diesterase activity is more than doubled. We have measured cAMP phosphodiesterase activity in various transformed cell lines.
Load more

Recommended publications
  • Effects of Aflatoxins Contaminating Food on Human Health - Magda Carvajal and Pável Castillo
    TROPICAL BIOLOGY AND CONSERVATION MANAGEMENT - – Vol.VII - Effects of Aflatoxins Contaminating Food on Human Health - Magda Carvajal and Pável Castillo EFFECTS OF AFLATOXINS CONTAMINATING FOOD ON HUMAN HEALTH Magda Carvajal and Pável Castillo Departamento de Botánica, Instituto de Biología, Universidad Nacional Autónoma de México. Ciudad Universitaria, Colonia Copilco, Delegación Coyoacán. 04510 México, D.F.(Institute of Biology, National Autonomous University of Mexico). Keywords: Mycotoxins, aflatoxins, cancer, mutagenesis, food contamination, DNA adducts, biomarkers, hepatic diseases, cirrhosis, hepatitis, toxicology, chemical mutations. Contents 1. Aflatoxins, production, occurrence, chemical structure 1.1 Definition of Aflatoxins 1.2. Aflatoxin Producing Fungi and Production Conditions 1.3. Occurrence 1.4. Chemical Structure and Types 1.5. Biological Properties 2. Biosynthetic pathway 2.1. Biotransformation of AFB1 3. Analytical methods for aflatoxin study 4. Aflatoxin metabolism 5. Toxic effects of aflatoxins on animal and human health 5.1 In Plants 5.2. In Animals 5.3. In Humans 6. Economic losses due to aflatoxin contamination 7. Control 7.1. Preventive Measures 7.2. Structural Degradation after Chemical Treatment 7.3. Modification of Toxicity by Dietary Chemicals 7.4. Detoxification 7.5. ChemosorbentsUNESCO – EOLSS 7.6. Radiation 8. Legislation 9. Conclusions Glossary SAMPLE CHAPTERS Bibliography Biographical Sketches Summary Aflatoxins (AF) are toxic metabolites of the moulds Aspergillus flavus, A. parasiticus and A. nomius. AF link to DNA, RNA and proteins and affect all the living kingdom, from viruses to man, causing acute or chronic symptoms, they are mutagens, hepatocarcinogens, and teratogens. ©Encyclopedia of Life Support Systems (EOLSS) TROPICAL BIOLOGY AND CONSERVATION MANAGEMENT - – Vol.VII - Effects of Aflatoxins Contaminating Food on Human Health - Magda Carvajal and Pável Castillo The impact of AF contamination on crops is estimated in hundreds of millions dollars.
    [Show full text]
  • Desmodus Rotundus) Blood Feeding
    toxins Article Vampire Venom: Vasodilatory Mechanisms of Vampire Bat (Desmodus rotundus) Blood Feeding Rahini Kakumanu 1, Wayne C. Hodgson 1, Ravina Ravi 1, Alejandro Alagon 2, Richard J. Harris 3 , Andreas Brust 4, Paul F. Alewood 4, Barbara K. Kemp-Harper 1,† and Bryan G. Fry 3,*,† 1 Department of Pharmacology, Biomedicine Discovery Institute, Faculty of Medicine, Nursing & Health Sciences, Monash University, Clayton, Victoria 3800, Australia; [email protected] (R.K.); [email protected] (W.C.H.); [email protected] (R.R.); [email protected] (B.K.K.-H.) 2 Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Cuernavaca, Morelos 62210, Mexico; [email protected] 3 Venom Evolution Lab, School of Biological Sciences, University of Queensland, St. Lucia, Queensland 4067, Australia; [email protected] 4 Institute for Molecular Biosciences, University of Queensland, St Lucia, QLD 4072, Australia; [email protected] (A.B.); [email protected] (P.F.A.) * Correspondence: [email protected] † Joint senior authors. Received: 20 November 2018; Accepted: 2 January 2019; Published: 8 January 2019 Abstract: Animals that specialise in blood feeding have particular challenges in obtaining their meal, whereby they impair blood hemostasis by promoting anticoagulation and vasodilation in order to facilitate feeding. These convergent selection pressures have been studied in a number of lineages, ranging from fleas to leeches. However, the vampire bat (Desmondus rotundus) is unstudied in regards to potential vasodilatory mechanisms of their feeding secretions (which are a type of venom). This is despite the intense investigations of their anticoagulant properties which have demonstrated that D.
    [Show full text]
  • Diagnosis of Clostridium Perfringens
    Received: January 12, 2009 J Venom Anim Toxins incl Trop Dis. Accepted: March 25, 2009 V.15, n.3, p.491-497, 2009. Abstract published online: March 31, 2009 Original paper. Full paper published online: August 31, 2009 ISSN 1678-9199. GENOTYPING OF Clostridium perfringens ASSOCIATED WITH SUDDEN DEATH IN CATTLE Miyashiro S (1), Baldassi L (1), Nassar AFC (1) (1) Animal Health Research and Development Center, Biological Institute, São Paulo, São Paulo State, Brazil. ABSTRACT: Toxigenic types of Clostridium perfringens are significant causative agents of enteric disease in domestic animals, although type E is presumably rare, appearing as an uncommon cause of enterotoxemia of lambs, calves and rabbits. We report herein the typing of 23 C. perfringens strains, by the polymerase chain reaction (PCR) technique, isolated from small intestine samples of bovines that have died suddenly, after manifesting or not enteric or neurological disorders. Two strains (8.7%) were identified as type E, two (8.7%) as type D and the remainder as type A (82.6%). Commercial toxoids available in Brazil have no label claims for efficacy against type E-associated enteritis; however, the present study shows the occurrence of this infection. Furthermore, there are no recent reports on Clostridium perfringens typing in the country. KEY WORDS: Clostridium perfringens, iota toxin, sudden death, PCR, cattle. CONFLICTS OF INTEREST: There is no conflict. CORRESPONDENCE TO: SIMONE MIYASHIRO, Instituto Biológico, Av. Conselheiro Rodrigues Alves, 1252, Vila Mariana, São Paulo, SP, 04014-002, Brasil. Phone: +55 11 5087 1721. Fax: +55 11 5087 1721. Email: [email protected]. Miyashiro S et al.
    [Show full text]
  • Venom Proteomics and Antivenom Neutralization for the Chinese
    www.nature.com/scientificreports OPEN Venom proteomics and antivenom neutralization for the Chinese eastern Russell’s viper, Daboia Received: 27 September 2017 Accepted: 6 April 2018 siamensis from Guangxi and Taiwan Published: xx xx xxxx Kae Yi Tan1, Nget Hong Tan1 & Choo Hock Tan2 The eastern Russell’s viper (Daboia siamensis) causes primarily hemotoxic envenomation. Applying shotgun proteomic approach, the present study unveiled the protein complexity and geographical variation of eastern D. siamensis venoms originated from Guangxi and Taiwan. The snake venoms from the two geographical locales shared comparable expression of major proteins notwithstanding variability in their toxin proteoforms. More than 90% of total venom proteins belong to the toxin families of Kunitz-type serine protease inhibitor, phospholipase A2, C-type lectin/lectin-like protein, serine protease and metalloproteinase. Daboia siamensis Monovalent Antivenom produced in Taiwan (DsMAV-Taiwan) was immunoreactive toward the Guangxi D. siamensis venom, and efectively neutralized the venom lethality at a potency of 1.41 mg venom per ml antivenom. This was corroborated by the antivenom efective neutralization against the venom procoagulant (ED = 0.044 ± 0.002 µl, 2.03 ± 0.12 mg/ml) and hemorrhagic (ED50 = 0.871 ± 0.159 µl, 7.85 ± 3.70 mg/ ml) efects. The hetero-specifc Chinese pit viper antivenoms i.e. Deinagkistrodon acutus Monovalent Antivenom and Gloydius brevicaudus Monovalent Antivenom showed negligible immunoreactivity and poor neutralization against the Guangxi D. siamensis venom. The fndings suggest the need for improving treatment of D. siamensis envenomation in the region through the production and the use of appropriate antivenom. Daboia is a genus of the Viperinae subfamily (family: Viperidae), comprising a group of vipers commonly known as Russell’s viper native to the Old World1.
    [Show full text]
  • Anti‑Inflammatory Effect of Bee Venom in an Allergic Chronic Rhinosinusitis Mouse Model
    6632 MOLECULAR MEDICINE REPORTS 17: 6632-6638, 2018 Anti‑inflammatory effect of bee venom in an allergic chronic rhinosinusitis mouse model SEUNG-HEON SHIN1, MI-KYUNG YE1, SUNG-YONG CHOI1 and KWAN-KYU PARK2 Departments of 1Otolaryngology-Head and Neck Surgery, and 2Pathology, School of Medicine, Catholic University of Daegu, Daegu 42472, Republic of Korea Received October 27, 2017; Accepted February 28, 2018 DOI: 10.3892/mmr.2018.8720 Abstract. Bee venom (BV) has long been used as fungal infection, and T-cell immune dysfunction (1-3). anti-inflammatory agent in traditional oriental medicine; Staphylococcus aureus produces proteins that act both as however, the effect of BV on chronic rhinosinusitis (CRS) is superantigens and toxins. Staphylococcal enterotoxin B (SEB) not commonly studied. The aim of the present study was to is commonly associated in the development of CRS with nasal determine the anti‑inflammatory effect of BV on an allergic polyp and specific IgE against SEB is more frequently detected CRS mouse model. An allergic CRS mouse model was in patients with nasal polyps than without nasal polyps (4). established following the administration of ovalbumin with Nasal exposure to SEB induce nasal polypoid lesion with Staphylococcus aureus enterotoxin B (SEB) into the nose. allergic rhinosinusitis in mice (5). Level of interleukin (IL)-5, A total of 0.5 or 5 ng/ml of BV were intranasally applied eotaxin in nasal lavage fluid (NLF) and number of secretory 3 times a week for 8 weeks. Histopathological alterations cells in nasal mucosa were increased in allergic rhinosinusitis were observed using hematoxylin and eosin, and Periodic acid model.
    [Show full text]
  • Cyanobacterial Toxins: Saxitoxins
    WHO/HEP/ECH/WSH/2020.8 Cyanobacterial toxins: saxitoxins Background document for development of WHO Guidelines for Drinking-water Quality and Guidelines for Safe Recreational Water Environments WHO/HEP/ECH/WSH/2020.8 © World Health Organization 2020 Some rights reserved. This work is available under the Creative Commons Attribution- NonCommercial-ShareAlike 3.0 IGO licence (CC BY-NC-SA 3.0 IGO; https://creativecommons.org/ licenses/by-nc-sa/3.0/igo). Under the terms of this licence, you may copy, redistribute and adapt the work for non-commercial purposes, provided the work is appropriately cited, as indicated below. In any use of this work, there should be no suggestion that WHO endorses any specific organization, products or services. The use of the WHO logo is not permitted. If you adapt the work, then you must license your work under the same or equivalent Creative Commons licence. If you create a translation of this work, you should add the following disclaimer along with the suggested citation: “This translation was not created by the World Health Organization (WHO). WHO is not responsible for the content or accuracy of this translation. The original English edition shall be the binding and authentic edition”. Any mediation relating to disputes arising under the licence shall be conducted in accordance with the mediation rules of the World Intellectual Property Organization (http://www.wipo.int/amc/en/ mediation/rules/). Suggested citation. Cyanobacterial toxins: saxitoxins. Background document for development of WHO Guidelines for drinking-water quality and Guidelines for safe recreational water environments. Geneva: World Health Organization; 2020 (WHO/HEP/ECH/WSH/2020.8).
    [Show full text]
  • Investigation of the Biochemical and Histological Changes Induced By
    Food and Nutrition Sciences, 2011, 2, 314-322 doi:10.4236/fns.2011.24045 Published Online June 2011 (http://www.scirp.org/journal/fns) Investigation of the Biochemical and Histological Changes Induced by Zearalenone Mycotoxin on Liver in Male Mice and the Protective Role of Crude Venom Extracted from Jellyfish Cassiopea Andromeda Madeha Al-Seeni1, Nagwa El-Sawi2, Soad Shaker3, Asma Al-Amoudi1 1Biochemistry Department, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia; 2Chemistry Department, Faculty of Science, Sohag University, Sohag, Egypt; 3Histology, Medicine Faculty, King Abdulaziz University, Jeddah, Saudi Arabia. Email: [email protected] Received February 1st, 2011; revised March 3rd, 2011; accepted March 8th, 2011. ABSTRACT Zearalenone (ZEN) is a non steroidal estrogenic mycotoxin produced by Fusarium species of fungi which contaminate human foods and animal feeds worldwide. In this study hepatotoxicity of ZEN was evaluated in mice by oral admini- stration of single and repeated doses of ZEN mycotoxin (2.7 mg/kg b.w.). The protective effect of crude venom ex- tracted from jellyfish Cassiopea andromeda was also assessed. Mice were divided into four groups (N = 10). G1: re- ceiving the toxin once and sacrificed 48 h later, G2: toxin administered twice for one week, G3: toxin administered twice a week for two weeks, G4: pretreated orally by a single dose of crude venom (1.78 mg/20 g) 24 hours prior to administration of ZEN twice a week for two weeks. Each treated group had its corresponding control which received 1% DMSO saline. ZEN treatment significantly increased alanine aminotransferase (ALT), aspartateaminotrnsferase (AST) and alkaline phosphatase (ALP) activities after 48 hours and two weeks, while ALT was also significantly in- creased after one week.
    [Show full text]
  • Snake Venom Detection Kit (SVDK)
    SVDK Template In non-urgent situations, serum or plasma may also be used. Other samples such as lymphatic fluid, tissue fluid or extracts may 8. Reading Colour Reactions be used. • Place the test strip on the template provided over page and observe each well continuously over the next 10 minutes whilst the colour develops. Any test sample used in the SVDK must be mixed with Yellow Sample Diluent (YSD-yellow lid), prior to introduction into the The first well to show visible colour, not including the positive control well, is assay. Samples mixed with YSD should be clearly labelled with the patient’s identity and the type of sample used. The volume of diagnostic of the snake’s venom immunotype – see interpretation below. YSD in each sample vial is sufficient to allow retesting of the sample or referral to a reference laboratory for further investigation. Well 1 Tiger Snake Immunotype Snake Venom Detection Kit (SVDK) Note: Strict adherence to the 10 minute observation period after addition of Tiger Snake Antivenom Indicated Detection and Identification of Snake Venom SAMPLE PREPARATION the Chromogen and Peroxide Solutions is essential. Slow development of 1. Prepare the Test Sample. colour in one or more wells after 10 minutes should not be interpreted ENZYME IMMUNOASSAY METHOD • Any test sample used in the SVDK must be mixed with Yellow Sample Diluent (YSD-yellow lid), prior to introduction as positive detection of snake venom. Well 2 Brown Snake Immunotype into the assay. INTERPRETATION OF RESULTS Brown Snake Antivenom Indicated Note: There is enough YSD in one vial to perform two snake venom detection tests.
    [Show full text]
  • Life Science Journal 2012;9(4) Http
    Life Science Journal 2012;9(4) http://www.lifesciencesite.com Biochemical and histological studies on the effect of the Patulin mycotoxin on male rats’ liver and treatment by crude venom extracted from jelly fish Nagwa M. El-Sawi*1,2, Hanaa M. Gashlan2, Sabry H. H. Younes1, Rehab F. Al-Massabi2 and S. Shaker3 1Chemistry Department, Faculty of Science, Sohag University, Sohag, 82524, Egypt 2Biochemistry Department, Faculty of Science, King Abdulaziz University, Jeddah 21551, Saudi Arabia 3Histology, Medicine Faculty King Abdulaziz University, Jeddah, 21551, Saudi Arabia [email protected] Abstract: Patulin mycotoxin on some biochemical parameters and histological changes on male rats' liver and effect of crude venom extracted from jelly fish Cassiopea Andromeda as a treatment. 50 Inbreeding weanling white male wistar lewis rats were divided randomly into 5 groups. Control group was gavage fed daily with distilled water; three treated groups were gavage fed daily dose with Patulin (0.2 mg/kg b.w.) for one, two and three weeks respectively. The last group was treated by Patulin for one week then injected intraperitoneally with single dose of crude venom (1.78 mg/20 g b.w.) for 24 hours according to LD50. Level of (AST) and (GGT) were increased significantly in serum of all treated groups compared with control group but level of (ALT) was increased significantly in treated group after one week only. Although the concentration of (TNF-α) was increased significantly and gradually in all treated groups, the concentration of ferritin was decreased significantly in treated three after three weeks only. Histopathological changes of rat liver coincided with biochemical changes.
    [Show full text]
  • Venom Toxins: Plausible Evolution from Digestive Enzymes1
    AMER. ZOOL., 23:427-430 (1983) Venom Toxins: Plausible Evolution from Digestive Enzymes1 ELAZAR KOCHVA,2 ORA NAKAR, AND MICHAEL OVADIA Department of Zoology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel SYNOPSIS Some hydrolytic enzymes are common to the pancreas, the mammalian salivary glands and the snake venom glands. Phospholipase A, which is found in elapid and viperid venoms and in the mammalian pancreas, shows 29 common amino acid residues out of Downloaded from https://academic.oup.com/icb/article/23/2/427/302344 by guest on 23 September 2021 118-125 positions. Presynaptic neurotoxins and other venom toxins are usually composed of 2-3 units or subumts, one of which is a phospholipase. The Vipera palaestmae two- component toxin retains its lethality when the enzyme is replaced by heterologous venom phospholipases, but not by the pig pancreatic enzyme. This toxin is neutralized by a factor found in the blood serum of snakes, which binds to the phospholipase and inhibits its activity. The blood serum of snakes also neutralizes hemorrhagins and inhibits the protease activity of the venom. It is hypothesized that the developing venom glands first produced enzymes that were already secreted by the pancreas and against which inhibitors were present in the blood. These inhibitors facilitated the evolution of enzyme-based toxins by neutralizing any damaging substances that might have escaped from the venom glands. It has been suggested that enzymes pre- certain elapid venom enzymes (Table 1). ceded toxins during the evolution of the Phospholipases from Viperidae venoms are venom glands in snakes (Strydom, 1977; the most divergent not only in certain areas Gans, 1978) possibly aiding in the digestion of their sequence, but also in the presence of prey and preventing its putrefaction of a seven residue COOH-terminal exten- (Thomas and Pough, 1979; Pough and sion (Heinrikson et al., 1977).
    [Show full text]
  • Binding of Saxitoxin to Electrically Excitable Neuroblastoma Cells (Ion Transport/Scorpion Toxin/Batrachotoxin) WILLIAM A
    Proc. Natl. Acad. Sci. USA Vol. 75, No. 1, pp. 218-222, January 1978 Biochemistry Binding of saxitoxin to electrically excitable neuroblastoma cells (ion transport/scorpion toxin/batrachotoxin) WILLIAM A. CATTERALL* AND CYNTHIA S. MORROW* Laboratory of Biochemical Genetics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20014 Communicated by Bernhard Witkop, November 1, 1977 ABSTRACT Saxitoxin inhibits the action potential Na+ permeability by 22Na+ influx (9), and measurement of scorpion ionophore of electrically excitable neuroblastoma cells with a toxin binding (11) have been described in detail. Scorpion toxin KI of 3.7 nM. Binding experiments detect a single class of sat- was purified by ion exchange chromatography and appeared urable binding sites with KD = 3.9 nM and a binding capacity of 156 fmol/mg of cell protein (78 sites per ,.m2 of cell surface). to be homogeneous in gel electrophoresis and isoelectric fo- Saturable binding is completely inhibited by tetrodotoxin but cusing experiments (9). The purified toxin was iodinated in a is unaffected by scorpion toxin or batrachotoxin. No saturable lactoperoxidase-catalyzed reaction and the labeled derivatives binding is observed in cultures of clone N103, a variant neuro- were separated by ion exchange chromatography (11). Unla- blastoma clone lacking the action potential Na+ response. Thus, beled, monoiodo and diiodo derivatives are separated by this saxitoxin binds specifically to the action potential Na+ iono- phore in neuroblastoma cells. Comparison of saxitoxin and technique. Only diiodo scorpion toxin was used in these studies. scorpion toxin binding reveals that there are three saxitoxin Because the preparations used contained only diiodo scorpion receptor sites for each scorpion toxin receptor site.
    [Show full text]
  • Phoneutria Nigriventer Spider Toxin Pntx2-1 (Δ-Ctenitoxin-Pn1a) Is a Modulator of Sodium Channel Gating
    toxins Article Phoneutria nigriventer Spider Toxin PnTx2-1 (δ-Ctenitoxin-Pn1a) Is a Modulator of Sodium Channel Gating Steve Peigneur 1,2,*,† ID , Ana Luiza B. Paiva 3,†, Marta N. Cordeiro 3,Márcia H. Borges 3, Marcelo R. V. Diniz 3, Maria Elena de Lima 2,4 and Jan Tytgat 1,* 1 Toxicology and Pharmacology, University of Leuven (KU Leuven), Campus Gasthuisberg, P.O. Box 922, Herestraat 49, 3000 Leuven, Belgium 2 Laboratório de Venenos e Toxinas Animais, Dept de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais (UFMG), Belo Horizonte 31270-901, Brazil; [email protected] 3 Departamento de Pesquisa e Desenvolvimento, Fundação Ezequiel Dias, Minas Gerais, Belo Horizonte 30510-010, Brazil; [email protected] (A.L.B.P.); [email protected] (M.N.C.); [email protected] (M.H.B.); [email protected] (M.R.V.D.) 4 Programa de Pós-graduação em Ciências da Saúde, Biomedicina e Medicina, Instituto de Ensino e Pesquisa da Santa Casa de Belo Horizonte, Grupo Santa Casa de Belo Horizonte, Minas Gerais, Belo Horizonte 31270-901, Brazil * Correspondence: [email protected] or [email protected] (S.P.); [email protected] (J.T.); Tel.: +321-632-3404 (J.T.) † These two authors contribute equally to this work. Received: 26 July 2018; Accepted: 16 August 2018; Published: 21 August 2018 Abstract: Spider venoms are complex mixtures of biologically active components with potentially interesting applications for drug discovery or for agricultural purposes. The spider Phoneutria nigriventer is responsible for a number of envenomations with sometimes severe clinical manifestations in humans.
    [Show full text]