Effects of vitamin C polyphosphate on liver lipid peroxides and antioxidase activity in mice 3

Σχετικά έγγραφα
cus sinensis 3 50 (2) : , 2004 ZHOU Xian2Qing 1,2, XIE Meng2Xia 3, N IU Cui2J uan 1 33, SUN Ru2Yong 1 ν 2004 Acta Zoologica Sinica

Science of Sericulture

IL - 13 /IL - 18 ELISA PCR RT - PCR. IL - 13 IL - 18 mrna. 13 IL - 18 mrna IL - 13 /IL Th1 /Th2

[11].,, , 316 6, ,., 15.5%, 9.8%, 2006., IDF,, ,500, 2,830.,, ,200.,,, β, [12]. 90% 2,,,,, [13-15].,, [13,

The toxicity of three chitin synthesis inhibitors to Calliptamus italicus Othoptera Acridoidea

Inhibition of mushroom tyrosinase by flavonoid from Sorbus tianschanica Ruper in Xinjiang

, ) ( mg/ kg 4, C3

Αξιοποίηση Φυσικών Αντιοξειδωτικών στην Εκτροφή των Αγροτικών

8Q5SAC) 8Q5SAC UV2Vis 8500 ( ) ; PHS23C ) ;721 ( ) :1 4. ;8Q5SAC : molπl ;Britton2Robinson Q5SAC BSA Britton2Robinson,

Antimicrobial Ability of Limonene, a Natural and Active Monoterpene

LUO, Hong2Qun LIU, Shao2Pu Ξ LI, Nian2Bing

0. 35g kg ~ 2. 0cm 10min. Nar- 15μL 6mm

Approximation Expressions for the Temperature Integral

Mean bond enthalpy Standard enthalpy of formation Bond N H N N N N H O O O

T CD , 0199 gπkg BW soybean isoflavones groups. Another ten 22month2old female rats were used as a young

Octretide joint proton pump inhibitors in treating non-variceal gastrointestinal bleeding a Metaanalysis

Electrolyzed-Reduced Water as Artificial Hot Spring Water

TGFp FSH INH INH

Optimizing Microwave-assisted Extraction Process for Paprika Red Pigments Using Response Surface Methodology

( ) , ) , ; kg 1) 80 % kg. Vol. 28,No. 1 Jan.,2006 RESOURCES SCIENCE : (2006) ,2 ,,,, ; ;

Effects of soothing liver and invigorating spleen recipes of TCM on hepatic inflammatory cytokines of rats with nonalcoholic steatosishepatitis

Study of Ne w Chemiluminescence Technique Recognition of Sodium Azide by External Reference Method

Extract Isolation Purification and Identification of Polysaccharides from Exocarp of Unripe Fruits of Juglans mandshurica

Αξιοποίηση Φυσικών Αντιοξειδωτικών στην Εκτροφή των Αγροτικών

Χρηµατοδότηση: Ε.Ε., ΚΕΟΣΟΕ (Γ.Γ.Ε.Τ. ΠΕΝΕΔ 2001)

PNS mg kg - 1. Rb 1

Gro wth Properties of Typical Water Bloom Algae in Reclaimed Water

Studies on the Binding Mechanism of Several Antibiotics and Human Serum Albumin

BGP TRACP-5b BGP TRACP-5b P 0.05

High mobility group 1 HMG1

Differences in Contents of Neutral Aroma Components and Sensory Evaluation in Air-cured Burley Leaves from Different Curing Barns

Design and Fabrication of Water Heater with Electromagnetic Induction Heating

Cellular Physiology and Biochemistry

Comparison of carbon-sulfur and carbon-amine bond in therapeutic drug: -S-aromatic heterocyclic podophyllum derivatives display antitumor activity

Accumulation of Soil Arsenic by Panax notoginseng and Its Associated Health Risk

Study on the Strengthen Method of Masonry Structure by Steel Truss for Collapse Prevention

Supporting Information

PE CVVH PE+HP HP+CVVH HP+CVVH+PE CVVH. ENLAV of HCO - 3 ENLAV-HCO ± μmol /L HP+PE HP+CVVH HP+CVVH+PE

ΕΦΑΡΜΟΓΗ ΕΥΤΕΡΟΒΑΘΜΙΑ ΕΠΕΞΕΡΓΑΣΜΕΝΩΝ ΥΓΡΩΝ ΑΠΟΒΛΗΤΩΝ ΣΕ ΦΥΣΙΚΑ ΣΥΣΤΗΜΑΤΑ ΚΛΙΝΗΣ ΚΑΛΑΜΙΩΝ

ER-Tree (Extended R*-Tree)

Advance in Nanorealgar Studies

H επίδραση της γονικής παρουσίας και του παιχνιδιού σε επώδυνες διαδικασίες στα παιδιά

Preparation of Hydroxyapatite Coatings on Enamel by Electrochemical Technique

A Systematic Review of Procalcitonin for Early Detection of Septicemia of Newborn

ΓΕΩΠΟΝΙΚΟ ΠΑΝΕΠΙΣΤΗΜΙΟ ΑΘΗΝΩΝ

) ; GSP ) ;PXD g, 100 ml

Correction of chromatic aberration for human eyes with diffractive-refractive hybrid elements

*,* + -+ on Bedrock Bath. Hideyuki O, Shoichi O, Takao O, Kumiko Y, Yoshinao K and Tsuneaki G

Quick algorithm f or computing core attribute

Comparison of nutrient components in muscle of wild and farmed groups of Myxocyprinus asiaticus

ΤΕΧΝΟΛΟΓΙΚΟ ΠΑΝΕΠΙΣΤΗΜΙΟ ΚΥΠΡΟΥ ΣΧΟΛΗ ΓΕΩΠΟΝΙΚΩΝ ΕΠΙΣΤΗΜΩΝ ΒΙΟΤΕΧΝΟΛΟΓΙΑΣ ΚΑΙ ΕΠΙΣΤΗΜΗΣ ΤΡΟΦΙΜΩΝ. Πτυχιακή εργασία

ΙΔΡΥΜΑ. Θεσσαλονίκη, ύλα

ΜΕΛΕΤΗ ΤΩΝ ΣΥΝΘΗΚΩΝ ΕΚΧΥΛΙΣΗΣ ΦΑΙΝΟΛΙΚΩΝ ΣΥΣΤΑΤΙΚΩΝ ΑΠΟ ΤΟ ΦΥΤΟ Sideritis raeseri ΜΕ ΤΗ ΧΡΗΣΗ ΜΙΚΡΟΚΥΜΑΤΩΝ

ΣΔΥΝΟΛΟΓΙΚΟ ΠΑΝΔΠΙΣΗΜΙΟ ΚΤΠΡΟΤ ΥΟΛΗ ΔΠΙΣΗΜΧΝ ΤΓΔΙΑ ΣΜΗΜΑ ΝΟΗΛΔΤΣΙΚΗ. Πηπρηαθή εξγαζία

2 PbO 2. Pb 3 O 4 Sn. Ti/SnO 2 -Sb 2 O 4 -CF/PbO x SnO 2 -Sb PbO 2. Sn-Sb 1:1. 1 h. Sn:Sb=10:1. PbO 2 - CeO 2 PbO 2. [8] SnO 2 +Sb 2 O 4 _

Physical and Chemical Properties of the Nest-site Beach of the Horseshoe Crab Rehabilitated by Sand Placement

High order interpolation function for surface contact problem

Supporting information. An unusual bifunctional Tb-MOF for highly sensing of Ba 2+ ions and remarkable selectivities of CO 2 /N 2 and CO 2 /CH 4

Βιοδείκτες οξειδωτικού στρες

Μέτρηση της Ρυθµικής Ικανότητας σε Μαθητές Γυµνασίου που Ασχολούνται µε Αθλητικές ραστηριότητες Συνοδευµένες ή Όχι από Μουσική

Influence of Flow Rate on Nitrate Removal in Flow Process

E#ects of Drying on Bacterial Activity and Iron Formation in Acid Sulfate Soils

FENXI HUAXUE Chinese Journal of Analytical Chemistry. Savitzky-Golay. n = SG SG. Savitzky-Golay mmol /L 5700.

,,, (, ) , ;,,, ; -

Uncovering the impact of capsule shaped amine-type ligands on. Am(III)/Eu(III) separation

Apr Vol.26 No.2. Pure and Applied Mathematics O157.5 A (2010) (d(u)d(v)) α, 1, (1969-),,.

ΠΑΝΕΠΙΣΤΗΜΙΟ ΘΕΣΣΑΛΙΑΣ

GDNF 220YL) 2,52. (glial cell line2de2. 015d 15d rived neurotrophic factor GDNF) 1993 Lin B49 (L15) (FBS) (poly2l2lysin, pll) (laminin, LN) GDNF GDNF

Nguyen Hien Trang* **

J. of Math. (PRC) 6 n (nt ) + n V = 0, (1.1) n t + div. div(n T ) = n τ (T L(x) T ), (1.2) n)xx (nt ) x + nv x = J 0, (1.4) n. 6 n

ΤΕΧΝΟΛΟΓΙΚΟ ΠΑΝΕΠΙΣΤΗΜΙΟ ΚΥΠΡΟΥ ΤΜΗΜΑ ΝΟΣΗΛΕΥΤΙΚΗΣ

ph Maillard MRPs maillard reaction products Maillard ph kDa Maillard Maillard DOI /j. issn

Studies on Synthesis and Biological Activities of 2( 1 H21,2,42 Triazol212yl)2 2Arylthioethyl Substituted Phenyl Ketones

ΚΕΡΚΙΔΙΚΗ ΚΑΙ ΜΗΡΙΑΙΑ ΠΡΟΣΠΕΛΑΣΗ ΣΕ ΣΤΕΦΑΝΙΟΓΡΑΦΙΕΣ ΚΑΙ ΑΓΓΕΙΟΠΛΑΣΤΙΚΕΣ. ΜΙΑ ΣΥΓΚΡΙΤΙΚΗ ΜΕΛΕΤΗ

D-Glucosamine-derived copper catalyst for Ullmann-type C- N coupling reaction: theoretical and experimental study

MSM Men who have Sex with Men HIV -

N 2 O 15 N NH + NH 3 Rittenberg mg mmol L - 1 CuSO mg. 10 mol L. Pre-Concentration device PreCon

Metal-free Oxidative Coupling of Amines with Sodium Sulfinates: A Mild Access to Sulfonamides

Mitomycin C application for the prevention of postoperative synechiae formation at the anterior commissure.

ACTA CHINESE MEDICINE. diabetic nephropathies DN 24. urine protein quantitation in 24 hours 24hUTP serum creatinine Scr

Varietal Differences in Response of Main Root Traits to Nitrogen Application Time in Rice ( Oryza sativa L. )

Supporting Information. Asymmetric Binary-acid Catalysis with Chiral. Phosphoric Acid and MgF 2 : Catalytic

Γεωπονικό Πανεπιςτήμιο Αθηνών Τμήμα Αξιοποίηςησ Φυςικών Πόρων και Γεωργικήσ Μηχανικήσ

Application of Statistical Process Control in Pretreatment Production Process of Gardenia jasminoides

The pathway of the ozonation of 2,4,62trichlorophenol in aqueous solution

(Fenneropenaeus chinensis)

Relationship between Connective Tissue Diseases and Thyroid Diseases

Rapid determination of soluble reactive silicate in seawater by flow injection analysis with spectrophotometric detection and its application

Comparison of Evapotranspiration between Indigenous Vegetation and Invading Vegetation in a Bog

Congruence Classes of Invertible Matrices of Order 3 over F 2

Παιδιατρική ΒΟΡΕΙΟΥ ΕΛΛΑΔΟΣ, 23, 3. Γ Παιδιατρική Κλινική, Αριστοτέλειο Πανεπιστήμιο, Ιπποκράτειο Νοσοκομείο Θεσσαλονίκης, 2

«Συντήρηση αχλαδιών σε νερό. υπό την παρουσία σπόρων σιναπιού (Sinapis arvensis).»

Reaction of a Platinum Electrode for the Measurement of Redox Potential of Paddy Soil

Study on Purification Technology and Antioxidant Activity of Total Flavonoid from Eriobotryae Folium

Free Radical Initiated Coupling Reaction of Alcohols and. Alkynes: not C-O but C-C Bond Formation. Context. General information 2. Typical procedure 2

No. 7 Modular Machine Tool & Automatic Manufacturing Technique. Jul TH166 TG659 A

17 min R A (2009) To probe into the thermal property the mechanism of the thermal decomposition and the prospective

27 7 Vol. 27 No CHINESE JOURNAL OF APPLIED CHEMISTRY July CV SEM EIS DTD. MCMB / 0. 01% DTD MCMB /Li. 300 ma h /g 350 ma h /g

Μελέτη της έκφρασης του ογκοκατασταλτικού γονιδίου Cyld στον καρκίνο του μαστού

Supplementary information:

Transcript:

50 (3) :370-374 2004 A cta Zoologica S inica C 3 33 100054 C 4 24 35 % C 0 500 2 500 5 000 mg/ kg 4 C C C 2 500 5 000 mg/ kg C 500 mg/ kg ; C 5 000 mg/ kg C C [ 50 (3) : 370-374 2004 ] C Effects of vitamin C polyphosphate on liver lipid peroxides and antioxidase activity in mice 3 ZHOU Xian2Qing 33 L I Sheng2Li WAN G Xiao2Hui Q IAO Xin Department of Laboratory Animal Sciences Capital Medical University Beijing 100054 China Abstract The study was conducted to investigate the effects of vitamin C polyphosphate on liver lipid peroxides (L PO) and antioxidase activity in mice. Twenty2four mice were fed with 35 % vitamin C polyphosphate supplementation at the dosages of 0 500 2 500 5 000 mg/ kg diets respectively for 4 weeks. Livers were collected to analyze the content of L PO and the activities of the superoxide dismutases ( SOD) catalase ( CA T) and glutathione peroxidase ( GSH2Px). The L PO and SOD were determined with thiobarbituric acid reaction and nitrite2form method respectively ; and the CA T and GSH2Px with colorimetry method. The results showed that vitamin Cpolyphosphate had no significant effects on liver L PO levels in mice although there was a decreasing tendency with an increase of vitamin C polyphosphate levels. The ac2 tivities of SOD in the two groups supplemented vitamin C polyphosphate at 2 500 and 5 000 mg/ kg diets were significant2 ly higher than those of both control groups and the group supplemented vitamin C polyphosphate at 500 mg/ kg. The ac2 tivities of CA T in the two groups supplemented vitamin C polyphosphate at 2 500 and 5 000 mg/ kg diets were higher than that of control group. The activity of GSH2Px in the groups supplemented vitamin C polyphosphate at 5 000 mg/ kg diets was notably higher than those of other three groups. These results suggest that vitamin C polyphosphate can improve the antioxidase activity while the requirement of vitamin C polyphosphate for improving different antioxidase activity varies [ Acta Zoologica Sinica 50 (3) : 370-374 2004 ]. Key words Mice Vitamin C polyphosphate Lipid peroxide Superoxide distutase Catalase Glutathione peroxidase C ( Ozt urk2urek C et al. 2001) C ( 2003211203 2004201217 3 (2002 KJ085) ( This research was funded by the grant (No. 2002 KJ085) from Educational Committee of Bei2 jing) 33 (Corresponding author). ν 2003 Acta Zoologica Sinica E2mail : xianqingzhou @yahoo1com1cn

3 : C 371 2000) (Oz2 112 turk2urek et al. 2001) 11211 C3 C4 ( 1 9 2003) C E (W V) D Y892 C E ( 2004) 1 200 r/ min 4 800 g 15 min 4 (Sahin et al. 2002a) C 11212 Lowry et al. ( 1951) (Smith et al. 2002) DNA (Lutsenko et al. 2002) Kaikkonen et al. (2001) C 11213 ; (1990) C 754 532 nm OD ( 2002) 11214 Sinha C (1972) 754 570 nm OD C 11215 Oyanagui ; (1984) (1996) Ξ 754 550 C nm OD 1 Paglia and Valentine (1967) Lawrence and Burk 111 (1976) ICR 754 340 nm OD 24 3-4 SPSS 16-22 g 24 4 6 L SD 26 cm 15 cm 15 cm 0105 21 1 4 Felin2 754 500 nm OD 11216 2 C : 211 2813 % 613 % 4816 % C 219 % 8 % 519 % 15124 KJ / ( F 3 20 = g 4 C 35 % C 11934 P > 0105) L SD 0 500 2 500 5 000 C mg/ kg C ( 1 : A) - 20 212 C 4 ( F 3 20 = 81356 P < 0105) C 2 500 5 000 mg/ kg Ξ 1996... 63-67. Zhang RJ 1996. Effects of trace elements2zinc and manganese on immune function and affecting mechanism in broiler. Ph. D. Agricultural University 63-67. Thesis. China

72 3 50 C C 500 mg/ kg ( P < 0105) ( 1 : B) 213 (L PO) C ( F 3 20 = 4164 P < 0105) L SD C 2 500 5 000 mg/ kg C ( P < 0105) ( 1 : C) 214 2001) C (Sahin et al. 2002b) Huang et al. (2002) ( F 3 18 = P < 0105) C 5 000 mg/ kg C E C ( P < 0105) ( 1 : D) 3 311 C ( 2000) C ( Zhang et al. C (Lykkesfeldt 2002) 1 C ( ) A. B. C. D. LSD ( P < 0105) Fig. 1 Effects of vitamin C polyphosphate on levels of liver lipid peroxides and antioxidase activity in mice ( Mean S E) A. Lipid peroxides. B. Activity of superoxide dismutase. C. Activity of catalase. D. Activity of glutathione peroxidase. LSD multiple range test the values with different superscript letter are significant different among different dose groups ( P < 0105).

3 : C 373 C 4 C E C ( Shankaran et al. 2001 ; C Smith et al. 1999) ( Shankaran et al. 2001 ; Smith et al. 1999) ; ( 2002) C 312 C ( References) C 2 500 5 000 mg/ kg C 5 000 mg/ kg C C ( 2002) vitamin C lowers lipid peroxidation E C ( 2003) C O2-265 - 275. ; (Drewa et al. 1998 ; Smith et al. 1999) ; 567-574. ( 1994) C Drewa G Jakobczyk M Araszkiewicz A 1998. A role of free radicals in schizophrenia. Med. Sci. Monit. 4 : 1 111-1 115. Du WD Shen DM Huang CJ Zhou YQ 2002. The effect of high2 dose vitamin C on lipid peroxidization injury in patient with acute pancreatitis. J. Clin. Digest. Disease 14 (5) : 201-203 ( In Chi2 nese). Feng B Shi GS He M 1994. The changes of lipid peroxides super2 oxide dismutases and glutathione peroxidase in old and young mice. Chin. J. Lab. Ani. Sci. 4 (1) : 60-61 ( In Chinese). Huang HY Appel LJ Croft KD Miller ER Mori TA Puddey IB 2002. Effects of vitamin C and vitamin E on i n vivo lipid peroxida2 tion : results of a randomized controlled trial. Am. J. Clin. Nutr. 76 (3) : 549-555. Kaikkonen J Porkkala2Sarataho E Morrow JD Roberts LJ Nyysso2 nen K Salonen R Tuomainen TP Ristonmaa U Poulsen HE Salonen J T 2001. Supplementation with vitamin E but not with i n vivo in mildly hyperc2 holesterolemic men. Free. Radic. Res. 35 (6) : 967-978. Lawrence RA Burk RF 1976. Glutathione peroxidase activity in sele2 nium2dificient rat liver. Biochem. Biophys. Res. Commun. 71 : 952-958. Liu YM Zhng J Zeng L Liu XQ Li J Z Qiu SQ Cai HB Long J X 2003. Effects of vitamin E combined with vitamin C on lipid perox2 idation and antioxidant enzyme of quartz exposed alveolar macrophage. Industrial Hygiene and Occupation Disease 29 ( 1) : 23-25 ( In Chinese). Lowery OH Rosebrough NJ Farr AL Randall RJ 1951. Protein measurement with the folin phenol reagent. J. Biol. Chem. 193 : Lutsenko EA Carcamo J M Golde DW 2002. Vitamin C prevents DNA mutation induced by oxidative stress. J. Biol. Chem. 277 (19) : 16 895-16 899. Lykkesfeldt J 2002. Increased oxidative damage in vitamin C deficiency is accompanied by induction of ascorbic acid recycling capacity in young but not mature guinea pigs. Free. Radic. Res. 36 ( 5) : Ma WQ Yang LB Yang XM Wang XF Li SY Zhang Y Wu XQ Liu GX 2002. Effect of ascorbic acid on level of SOD in brain of rates under acute stress. Chin. J. Mental Hygiene 16 ( 12 ) : 809-810 ( In Chinese). Oyanagui Y 1984. Reevalution of assay methods and establishinent of kit for superoxide dismutase activity. Anal. Biochem. 142 : 290-296. Ozturk2Urek R Bozkaya LA Tarhan L 2001. The effects of some

74 3 50 antioxidant vitamin2 and trace element2supplemented diets on activi2 ties of SOD CAT GSH2Px and L PO levels in chicken tissues. Cell Biochem. Funct. 19 : 125-132. Paglia DE Valentine WN 1967. Studies on the quantitative and quali2 tative characterization of erythrocyte glutathione peroxidase. J. Lab. Clin. Med. 70 : 158-169. Sahin K Kucuk O Sahin N Sari M 2002a. Effects of vitamin C and vitamin E on lipid peroxidation status serum hormone metabo2 lite and mineral concentrations of Japanese quails reared under heat stress (34 ). Int. Vitam. Nutr. Res. 72 (2) : 91-100. Sahin K Sahin N Yaralioglu S 2002b. Effects of vitamin C and vita2 min E on lipid peroxidation blood serum metabolites and mineral concentrations of laying hens reared at high ambient temperature. Biol. Trace. Elem. Res. 85 (1) : 35-45. Shankaran M Yamamoto B K Gudelsky GA 2001 Ascorbic acid pre2 vents 3 42methylenedioxy methamphetamine ( MDMA) induced hydroxyl radical formation and the behavioral neurochemical conse2 quences of the depletion of brain 52HT. Synapse 40 : 55-64. Sinha A K 1972. Colorimetric assay of catalase. Anal. Biochem. 47 : 389-394. Smith AR Visioli F Hagen TM 2002. Vitamin C matters : increased oxidative stress in cultured human aortic endothelial cells without supplemental ascorbic acid. Faseb. J. 16 (9) : 1 102-1 104. Smith KL Kapoor R Felts PA 1999. Demyelination : the role of re2 active oxygen and nitrogen species. Brain Pathology 9 : 69-92. Wang M Y Wei SZ Lu LD Jiang LL 2000. Fuzzy integrative assess2 ment of the influences of antioxidant eenzyme activities on the con2 tent of lipid peroxides in rat liver. J. Math. Med. 13 (5) : 450-451. Wen J Lin J H Gao YQ 2002. Effect of dietary vitamin C supple2 mentation on production performance and the capacity of biosynthe2 sis of vitamin C in broilers under heat stress. Acta Veterinariaet Zootechnica Sinica 31 (6) : 497-502 ( In Chinese). Xiang R Wang DN 1990. The improvement of the method spec2 trophotometer on lipid peroxides thiobarbituric acid. Prog. Biochem. Biophy. 17 (3) : 241-242 ( In Chinese).. Zhang QS Dong GX Zhang SH 2002. Protective effects of vitamin C on the cultured human umbilical vein endothelial cell injured by lipid peroxidation. Chinese Journal of Arteriosclerosis 10 ( 3) : 225-227 ( In Chinese). Zhang QJ Li T Zhan H Xin YM 2001. Inhibitory effects of tea polyphenols and vitamin C on lipid peroxidation induced by FeSO 4 2 cysteine in isolated human plasma and carbon tetrachloride2induced liver free radical injury in mice. Space. Med. Med. Eng. (Bei2 jing) 14 (1) : 50-53. Zhou XQ Niu CJ Sun RY 2003. Effects of vitamin C and acid2stress on serum complement C3 and C4 level in juvenile soft2shelled turtles Pelodiscus sinensis. Acta Zool. Sin. 49 (6) : 769-774 ( In Chi2 nese). Zhou XQ Xie MX Niu CJ Sun R Y 2004. Effects of the combination of vitamin C and E on growth liver vitamin C and E and levels of serum cortisol in stressed and non2stressed juvenile soft2shelled tur2 tles Pelodiscus sinensis. Acta Zool. Sin. 50 (2) : 158-164 ( In Chinese). 2002. C. 14 (5) : 201-203. 1994.. 4 (1) : 60-61. 2003. E C. 29 (1) : 23-25. 2002. C. 16 (12) : 809-810. 2000.. 13 (5) : 450-451. 2002. C C. 31 (6) : 497-502. 1990... 17 (3) : 241-242. 2002. C. 10 (3) : 225-227 2003. C C3 C4. 49 (6) : 769-774. 2004. C E C E. 50 (2) : 158-164.