Tom 8, Nr 2 (2016)
Artykuły
Opublikowany online: 2016-12-08
Wyświetlenia strony
2089
Wyświetlenia/pobrania artykułu
10203
Wpływ mikrobioty jelitowej na zaburzenia metaboliczne i otyłość — punkt widzenia internisty i dietetyka
Gastroenterologia Kliniczna 2016;8(2).
Streszczenie
Otyłość staje się jednym z najpoważniejszych problemów zdrowotnych większości społczeństw na świecie. Nadal poszukuje się przyczyn takiego stanu, a zwłaszcza modyfikowalnych czynników wpływających na równowagę energetyczną organizmu. W ostatnim czasie coraz więcej przeprowadzonych badań umożliwia lepsze zrozumienie funkcji mkirobioty jelitowej oraz jej wpływu na metabolizm i magazynowanie energii w organizmie gospodarza. W niniejszej pracy przedstawiono aktualną wiedzę na temat roli mikrobioty jelitowej w patogenezie otyłości, insulinooporności, cukrzycy typu 2, zaburzeń lipidowych i niealkoholowej stłuszczeniowej choroby wątroby. Mikrobiota zdecydowanie różni się u osób otyłych i szczupłych; u osób otyłych charakteryzuje się głównie spadkiem różnorodności, zmniejszeniem liczby bakterii komensalnych o właściwościach przeciwzapalnych i wzrostem liczby drobnoustrojów patogennych. Generuje to ekspresję odmiennych szlaków metabolicznych, co może stanowić jeden z czynników etiologicznych otyłości i chorób jej towarzyszących. Pro- i prebiotykoterapia jest aktualnie wykorzystywana w celach rekompozycji ekosystemu jelitowego.
Słowa kluczowe: otyłośćmikrobiotazaburzenia metaboliczneprobiotykiprebiotyki
Referencje
- Swinburn B, Dietz W, Kleinert S. A Lancet Commission on obesity. Lancet. 2015; 386(10005): 1716–1717.
- World Health Organization. Obesity and overweight, updated January 2015. http://www.who.int/mediacentre/factsheets/ fs311/en/ (05.11.2015).
- Festi D, Schiumerini R, Eusebi LH, et al. Gut microbiota and metabolic syndrome. World J Gastroenterol. 2014; 20(43): 16079–16094.
- DiBaise JK, Zhang H, Crowell MD, et al. Gut microbiota and its possible relationship with obesity. Mayo Clin Proc. 2008; 83(4): 460–469.
- Turnbaugh PJ, Ley RE, Mahowald MA, et al. An obesity-associated gut microbiome with increased capacity for energy harvest. Nature. 2006; 444(7122): 1027–1031.
- Marchesi J, Shanahan F. The normal intestinal microbiota. Curr Opin Infect Dis. 2007; 20(5): 508–513.
- Frank DN, St Amand AL, Feldman RA, et al. Molecular-phylogenetic characterization of microbial community imbalances in human inflammatory bowel diseases. Proc Natl Acad Sci U S A. 2007; 104(34): 13780–13785.
- Turnbaugh PJ, Ley RE, Hamady M, et al. The human microbiome project. Nature. 2007; 449(7164): 804–810.
- Favier CF, Vaughan EE, De Vos WM, et al. Molecular monitoring of succession of bacterial communities in human neonates. Appl Environ Microbiol. 2002; 68(1): 219–226.
- Fukuda S, Ohno H. Gut microbiome and metabolic diseases. Semin Immunopathol. 2014; 36(1): 103–114.
- Hooper LV, Littman DR, Macpherson AJ. Interactions between the microbiota and the immune system. Science. 2012; 336(6086): 1268–1273.
- Jia W, Li H, Zhao L, et al. Gut microbiota: a potential new territory for drug targeting. Nat Rev Drug Discov. 2008; 7(2): 123–129.
- Neu J, Douglas-Escobar M, Lopez M. Microbes and the developing gastrointestinal tract. Nutr Clin Pract. 2007; 22(2): 174–182.
- Cani PD, Amar J, Iglesias MA, et al. Metabolic endotoxemia initiates obesity and insulin resistance. Diabetes. 2007; 56(7): 1761–1772.
- Turnbaugh PJ, Bäckhed F, Fulton L, et al. Diet-induced obesity is linked to marked but reversible alterations in the mouse distal gut microbiome. Cell Host Microbe. 2008; 3(4): 213–223.
- Marlicz W, Zawada I, Starzyńska T. Zespół nadwrażliwego jelita — nadwrażliwe jelito czy nadwrażliwy umysł? Polski Merkuriusz Lekarski. 2012; XXXII(187): 64–69.
- Bäckhed F, Ding H, Wang T, et al. The gut microbiota as an environmental factor that regulates fat storage. Proc Natl Acad Sci U S A. 2004; 101(44): 15718–15723.
- Bajzer M, Seeley RJ. Physiology: obesity and gut flora. Nature. 2006; 444(7122): 1009–1010.
- Flint HJ, Bayer EA, Rincon MT, et al. Polysaccharide utilization by gut bacteria: potential for new insights from genomic analysis. Nat Rev Microbiol. 2008; 6(2): 121–131.
- Harris K, Kassis A, Major G, et al. Is the gut microbiota a new factor contributing to obesity and its metabolic disorders? J Obes. 2012; 2012: 879151.
- Schwiertz A, Taras D, Schäfer K, et al. Microbiota and SCFA in lean and overweight healthy subjects. Obesity (Silver Spring). 2010; 18(1): 190–195.
- Samuel BS, Shaito A, Motoike T, et al. Effects of the gut microbiota on host adiposity are modulated by the short-chain fatty-acid binding G protein-coupled receptor, Gpr41. Proc Natl Acad Sci U S A. 2008; 105(43): 16767–16772.
- Tolhurst G, Heffron H, Lam YuS, et al. Short-chain fatty acids stimulate glucagon-like peptide-1 secretion via the G-protein-coupled receptor FFAR2. Diabetes. 2012; 61(2): 364–371.
- Hooper LV, Wong MH, Thelin A, et al. Molecular analysis of commensal host-microbial relationships in the intestine. Science. 2001; 291(5505): 881–884.
- Grudell ABM, Camilleri M. The role of peptide YY in integrative gut physiology and potential role in obesity. Curr Opin Endocrinol Diabetes Obes. 2007; 14(1): 52–57.
- Nicholson JK, Holmes E, Kinross J, et al. Host-gut microbiota metabolic interactions. Science. 2012; 336(6086): 1262–1267.
- Gao Z, Yin J, Zhang J, et al. Butyrate improves insulin sensitivity and increases energy expenditure in mice. Diabetes. 2009; 58(7): 1509–1517.
- Stappenbeck TS, Hooper LV, Gordon JI. Developmental regulation of intestinal angiogenesis by indigenous microbes via Paneth cells. Proc Natl Acad Sci U S A. 2002; 99(24): 15451–15455.
- Mandard S, Zandbergen F, van Straten E, et al. The fasting-induced adipose factor/angiopoietin-like protein 4 is physically associated with lipoproteins and governs plasma lipid levels and adiposity. J Biol Chem. 2006; 281(2): 934–944.
- Ley RE, Bäckhed F, Turnbaugh P, et al. Obesity alters gut microbial ecology. Proc Natl Acad Sci USA. 2005; 102(31): 11070–11075.
- Ley RE, Turnbaugh PJ, Klein S, et al. Microbial ecology: human gut microbes associated with obesity. Nature. 2006; 444(7122): 1022–1023.
- Turnbaugh PJ, Ridaura VK, Faith JJ, et al. The effect of diet on the human gut microbiome: a metagenomic analysis in humanized gnotobiotic mice. Sci Transl Med. 2009; 1(6): 614.
- Duncan SH, Lobley GE, Holtrop G, et al. Human colonic microbiota associated with diet, obesity and weight loss. Int J Obes (Lond). 2008; 32(11): 1720–1724.
- Samuel BS, Hansen EE, Manchester JK, et al. Genomic and metabolic adaptations of Methanobrevibacter smithii to the human gut. Proc Natl Acad Sci USA. 2007; 104(25): 10643–10648.
- Furet JP, Kong LC, Tap J, et al. Differential adaptation of human gut microbiota to bariatric surgery-induced weight loss: links with metabolic and low-grade inflammation markers. Diabetes. 2010; 59(12): 3049–3057.
- De Filippo C, Cavalieri D, Di Paola M, et al. Impact of diet in shaping gut microbiota revealed by a comparative study in children from Europe and rural Africa. Proc Natl Acad Sci USA. 2010; 107(33): 14691–14696.
- Hildebrandt MA, Hoffmann C, Sherrill-Mix SA, et al. High-fat diet determines the composition of the murine gut microbiome independently of obesity. Gastroenterology. 2009; 137(5): 1716–17124.
- David LA, Maurice CF, Carmody RN, et al. Diet rapidly and reproducibly alters the human gut microbiome. Nature. 2014; 505(7484): 559–563.
- Shen W, Gaskins HR, McIntosh MK. Influence of dietary fat on intestinal microbes, inflammation, barrier function and metabolic outcomes. J Nutr Biochem. 2014; 25(3): 270–280.
- Brinkworth GD, Noakes M, Clifton PM, et al. Comparative effects of very low-carbohydrate, high-fat and high-carbohydrate, low-fat weight-loss diets on bowel habit and faecal short-chain fatty acids and bacterial populations. Br J Nutr. 2009; 101(10): 1493–1502.
- Zhang C, Zhang M, Wang S, et al. Interactions between gut microbiota, host genetics and diet relevant to development of metabolic syndromes in mice. ISME J. 2010; 4(2): 232–241.
- Wright SD, Ramos RA, Tobias PS, et al. CD14, a receptor for complexes of lipopolysaccharide (LPS) and LPS binding protein. Science. 1990; 249(4975): 1431–1433.
- Jumpertz R, Le DS, Turnbaugh PJ, et al. Energy-balance studies reveal associations between gut microbes, caloric load, and nutrient absorption in humans. Am J Clin Nutr. 2011; 94(1): 58–65.
- Arumugam M, Raes J, Pelletier E, et al. MetaHIT Consortium. Enterotypes of the human gut microbiome. Nature. 2011; 473(7346): 174–180.
- Le Chatelier E, Nielsen T, Qin J, et al. MetaHIT consortium. Richness of human gut microbiome correlates with metabolic markers. Nature. 2013; 500(7464): 541–546.
- Membrez M, Blancher F, Jaquet M, et al. Gut microbiota modulation with norfloxacin and ampicillin enhances glucose tolerance in mice. FASEB J. 2008; 22(7): 2416–2426.
- Cani PD, Bibiloni R, Knauf C, et al. Changes in gut microbiota control metabolic endotoxemia-induced inflammation in high-fat diet-induced obesity and diabetes in mice. Diabetes. 2008; 57(6): 1470–1481.
- Amar J, Burcelin R, Ruidavets JB, et al. Energy intake is associated with endotoxemia in apparently healthy men. Am J Clin Nutr. 2008; 87(5): 1219–1223.
- Creely SJ, McTernan PG, Kusminski CM, et al. Lipopolysaccharide activates an innate immune system response in human adipose tissue in obesity and type 2 diabetes. Am J Physiol Endocrinol Metab. 2007; 292(3): E740–E747.
- Shi H, Kokoeva MV, Inouye K, et al. TLR4 links innate immunity and fatty acid-induced insulin resistance. J Clin Invest. 2006; 116(11): 3015–3025.
- Amar J, Serino M, Lange C, et al. D.E.S.I.R. Study Group. Involvement of tissue bacteria in the onset of diabetes in humans: evidence for a concept. Diabetologia. 2011; 54(12): 3055–3061.
- Zhang X, Shen D, Fang Z, et al. Human gut microbiota changes reveal the progression of glucose intolerance. PLoS One. 2013; 8(8): e71108.
- Lewis K, Lutgendorff F, Phan V, et al. Enhanced translocation of bacteria across metabolically stressed epithelia is reduced by butyrate. Inflamm Bowel Dis. 2010; 16(7): 1138–1148.
- Vrieze A, Van Nood E, Holleman F, et al. Transfer of intestinal microbiota from lean donors increases insulin sensitivity in individuals with metabolic syndrome. Gastroenterology. 2012; 143(4): 913–916.
- Alisi A, Ceccarelli S, Panera N, et al. Causative role of gut microbiota in non-alcoholic fatty liver disease pathogenesis. Front Cell Infect Microbiol. 2012; 2: 132.
- Mishra AK, Dubey V, Ghosh AR. Obesity: An overview of possible role(s) of gut hormones, lipid sensing and gut microbiota. Metabolism. 2016; 65(1): 48–65.
- Bäckhed F, Manchester JK, Semenkovich CF, et al. Mechanisms underlying the resistance to diet-induced obesity in germ-free mice. Proc Natl Acad Sci U S A. 2007; 104(3): 979–984.
- Bäckhed F, Crawford PA, O'Donnell D, et al. Postnatal lymphatic partitioning from the blood vasculature in the small intestine requires fasting-induced adipose factor. Proc Natl Acad Sci U S A. 2007; 104(2): 606–611.
- Vreugdenhil ACE, Rousseau CH, Hartung T, et al. Lipopolysaccharide (LPS)-binding protein mediates LPS detoxification by chylomicrons. J Immunol. 2003; 170(3): 1399–1405.
- Lefebvre P, Cariou B, Lien F, et al. Role of bile acids and bile acid receptors in metabolic regulation. Physiol Rev. 2009; 89(1): 147–191.
- Swann JR, Want EJ, Geier FM, et al. Systemic gut microbial modulation of bile acid metabolism in host tissue compartments. Proc Natl Acad Sci U S A. 2011; 108 Suppl 1: 4523–4530.
- Thomas C, Gioiello A, Noriega L, et al. TGR5-mediated bile acid sensing controls glucose homeostasis. Cell Metab. 2009; 10(3): 167–177.
- Pappo I, Becovier H, Berry EM, et al. Polymyxin B reduces cecal flora, TNF production and hepatic steatosis during total parenteral nutrition in the rat. J Surg Res. 1991; 51(2): 106–112.
- Cope K, Risby T, Diehl AM. Increased gastrointestinal ethanol production in obese mice: implications for fatty liver disease pathogenesis. Gastroenterology. 2000; 119(5): 1340–1347.
- Zhu L, Baker SS, Gill C, et al. Characterization of gut microbiomes in nonalcoholic steatohepatitis (NASH) patients: a connection between endogenous alcohol and NASH. Hepatology. 2013; 57(2): 601–609.
- Dumas ME, Barton RH, Toye A, et al. Metabolic profiling reveals a contribution of gut microbiota to fatty liver phenotype in insulin-resistant mice. Proc Natl Acad Sci U S A. 2006; 103(33): 12511–12516.
- Buchman AL, Dubin MD, Moukarzel AA, et al. Choline deficiency: a cause of hepatic steatosis during parenteral nutrition that can be reversed with intravenous choline supplementation. Hepatology. 1995; 22(5): 1399–1403.
- Spencer MD, Hamp TJ, Reid RW, et al. Association between composition of the human gastrointestinal microbiome and development of fatty liver with choline deficiency. Gastroenterology. 2011; 140(3): 976–986.
- Rivera CA, Adegboyega P, van Rooijen N, et al. Toll-like receptor-4 signaling and Kupffer cells play pivotal roles in the pathogenesis of non-alcoholic steatohepatitis. J Hepatol. 2007; 47(4): 571–579.
- Miele L, Valenza V, La Torre G, et al. Increased intestinal permeability and tight junction alterations in nonalcoholic fatty liver disease. Hepatology. 2009; 49(6): 1877–1887.
- Amar J, Chabo C, Waget A, et al. Intestinal mucosal adherence and translocation of commensal bacteria at the early onset of type 2 diabetes: molecular mechanisms and probiotic treatment. EMBO Mol Med. 2011; 3(9): 559–572.
- Cani PD, Neyrinck AM, Fava F, et al. Selective increases of bifidobacteria in gut microflora improve high-fat-diet-induced diabetes in mice through a mechanism associated with endotoxaemia. Diabetologia. 2007; 50(11): 2374–2383.
- Lee HY, Park JH, Seok SH, et al. Human originated bacteria, Lactobacillus rhamnosus PL60, produce conjugated linoleic acid and show anti-obesity effects in diet-induced obese mice. Biochim Biophys Acta. 2006; 1761(7): 736–744.
- Lee K, Paek K, Lee HY, et al. Antiobesity effect of trans-10,cis-12-conjugated linoleic acid-producing Lactobacillus plantarum PL62 on diet-induced obese mice. J Appl Microbiol. 2007; 103(4): 1140–1146.
- Sato M, Uzu K, Yoshida T, et al. Effects of milk fermented by Lactobacillus gasseri SBT2055 on adipocyte size in rats. Br J Nutr. 2008; 99(5): 1013–1017.
- Kondo S, Xiao JZ, Satoh T, et al. Antiobesity effects of Bifidobacterium breve strain B-3 supplementation in a mouse model with high-fat diet-induced obesity. Biosci Biotechnol Biochem. 2010; 74(8): 1656–1661.
- Cano PG, Santacruz A, Trejo FM, et al. Bifidobacterium CECT 7765 improves metabolic and immunological alterations associated with obesity in high-fat diet-fed mice. Obesity (Silver Spring). 2013; 21(11): 2310–2321.
- Chen JJ, Wang R, Li Xf, et al. Bifidobacterium longum supplementation improved high-fat-fed-induced metabolic syndrome and promoted intestinal Reg I gene expression. Exp Biol Med (Maywood). 2011; 236(7): 823–831.
- Tomaro-Duchesneau C, Saha S, Malhotra M, et al. Effect of orally administered L. fermentum NCIMB 5221 on markers of metabolic syndrome: an in vivo analysis using ZDF rats. Appl Microbiol Biotechnol. 2014; 98(1): 115–126.
- Wang LX, Liu K, Gao DW, et al. Protective effects of two Lactobacillus plantarum strains in hyperlipidemic mice. World J Gastroenterol. 2013; 19(20): 3150–3156.
- Kim SW, Park KY, Kim B, et al. Lactobacillus rhamnosus GG improves insulin sensitivity and reduces adiposity in high-fat diet-fed mice through enhancement of adiponectin production. Biochem Biophys Res Commun. 2013; 431(2): 258–263.
- Fåk F, Bäckhed F. Lactobacillus reuteri prevents diet-induced obesity, but not atherosclerosis, in a strain dependent fashion in Apoe-/- mice. PLoS One. 2012; 7(10): e46837.
- Takemura N, Okubo T, Sonoyama K. Lactobacillus plantarum strain No. 14 reduces adipocyte size in mice fed high-fat diet. Exp Biol Med (Maywood). 2010; 235(7): 849–856.
- Aronsson L, Huang Y, Parini P, et al. Decreased fat storage by Lactobacillus paracasei is associated with increased levels of angiopoietin-like 4 protein (ANGPTL4). PLoS One. 2010; 5(9): e13087.
- Nerstedt A, Nilsson EC, Ohlson K, et al. Administration of Lactobacillus evokes coordinated changes in the intestinal expression profile of genes regulating energy homeostasis and immune phenotype in mice. Br J Nutr. 2007; 97(6): 1117–1127.
- Kadooka Y, Sato M, Imaizumi K, et al. Regulation of abdominal adiposity by probiotics (Lactobacillus gasseri SBT2055) in adults with obese tendencies in a randomized controlled trial. Eur J Clin Nutr. 2010; 64(6): 636–643.
- Sanchez M, Darimont C, Drapeau V, et al. Effect of Lactobacillus rhamnosus CGMCC1.3724 supplementation on weight loss and maintenance in obese men and women. Br J Nutr. 2014; 111(8): 1507–1519.
- Kadooka Y, Sato M, Ogawa A, et al. Effect of Lactobacillus gasseri SBT2055 in fermented milk on abdominal adiposity in adults in a randomised controlled trial. Br J Nutr. 2013; 110(9): 1696–1703.
- Barreto FM, Colado Simão AN, Morimoto HK, et al. Beneficial effects of Lactobacillus plantarum on glycemia and homocysteine levels in postmenopausal women with metabolic syndrome. Nutrition. 2014; 30(7-8): 939–942.
- Yadav H, Lee JH, Lloyd J, et al. Beneficial metabolic effects of a probiotic via butyrate-induced GLP-1 hormone secretion. J Biol Chem. 2013; 288(35): 25088–25097.
- Roberfroid M, Gibson GR, Hoyles L, et al. Prebiotic effects: metabolic and health benefits. Br J Nutr. 2010; 104 Suppl 2: S1–63.
- Neyrinck AM, Possemiers S, Druart C, et al. Prebiotic effects of wheat arabinoxylan related to the increase in bifidobacteria, Roseburia and Bacteroides/Prevotella in diet-induced obese mice. PLoS One. 2011; 6(6): e20944.
- Parnell JA, Reimer RA. Prebiotic fibres dose-dependently increase satiety hormones and alter Bacteroidetes and Firmicutes in lean and obese JCR:LA-cp rats. Br J Nutr. 2012; 107(4): 601–613.
- Everard A, Lazarevic V, Derrien M, et al. Responses of gut microbiota and glucose and lipid metabolism to prebiotics in genetic obese and diet-induced leptin-resistant mice. Diabetes. 2011; 60(11): 2775–2786.
- Cani PD, Neyrinck AM, Maton N, et al. Oligofructose promotes satiety in rats fed a high-fat diet: involvement of glucagon-like Peptide-1. Obes Res. 2005; 13(6): 1000–1007.
- Cani PD, Daubioul CA, Reusens B, et al. Involvement of endogenous glucagon-like peptide-1(7-36) amide on glycaemia-lowering effect of oligofructose in streptozotocin-treated rats. J Endocrinol. 2005; 185(3): 457–465.
- Cani PD, Possemiers S, Van de Wiele T, et al. Changes in gut microbiota control inflammation in obese mice through a mechanism involving GLP-2-driven improvement of gut permeability. Gut. 2009; 58(8): 1091–1103.
- Kellow NJ, Coughlan MT, Reid CM. Metabolic benefits of dietary prebiotics in human subjects: a systematic review of randomised controlled trials. Br J Nutr. 2014; 111(7): 1147–1161.
- Parnell JA, Reimer RA. Weight loss during oligofructose supplementation is associated with decreased ghrelin and increased peptide YY in overweight and obese adults. Am J Clin Nutr. 2009; 89(6): 1751–1759.
- Genta S, Cabrera W, Habib N, et al. Yacon syrup: beneficial effects on obesity and insulin resistance in humans. Clin Nutr. 2009; 28(2): 182–187.
- Dewulf EM, Cani PD, Claus SP, et al. Insight into the prebiotic concept: lessons from an exploratory, double blind intervention study with inulin-type fructans in obese women. Gut. 2013; 62(8): 1112–1121.
- Sotos M, Nadal I, Marti A, et al. Gut microbes and obesity in adolescents. Proceedings of the Nutrition Society. 2008; 67: E20.
- Nadal I, Santacruz A, Marcos A, et al. Shifts in clostridia, bacteroides and immunoglobulin-coating fecal bacteria associated with weight loss in obese adolescents. Int J Obes (Lond). 2009; 33(7): 758–767.
- Cotillard A, Kennedy SP, Kong LC, et al. ANR MicroObes consortium. Dietary intervention impact on gut microbial gene richness. Nature. 2013; 500(7464): 585–588.
- Zhang H, DiBaise JK, Zuccolo A, et al. Human gut microbiota in obesity and after gastric bypass. Proc Natl Acad Sci U S A. 2009; 106(7): 2365–2370.
- Li JV, Ashrafian H, Bueter M, et al. Metabolic surgery profoundly influences gut microbial-host metabolic cross-talk. Gut. 2011; 60(9): 1214–1223.
- Lips MA, de Groot GH, van Klinken JB, et al. Calorie restriction is a major determinant of the short-term metabolic effects of gastric bypass surgery in obese type 2 diabetic patients. Clin Endocrinol (Oxf). 2014; 80(6): 834–842.
- Ostrowska L, Marlicz W, Łoniewski I. Transplantacja mikroflory jelitowej w leczeniu otyłości i zaburzeń metabolicznych — metoda nadal ryzykowna i niepotwierdzona wynikami badań klinicznych. Forum Zaburzeń Metabolicznych 2013; 4. ; 161: 169.
- Dror T, Dickstein Y, Dubourg G, et al. Microbiota manipulation for weight change. Microb Pathog. 2016 [Epub ahead of print].
- Gøbel RJ, Larsen N, Jakobsen M, et al. Probiotics to adolescents with obesity: effects on inflammation and metabolic syndrome. J Pediatr Gastroenterol Nutr. 2012; 55(6): 673–678.
- Lee SJi, Bose S, Seo JG, et al. The effects of co-administration of probiotics with herbal medicine on obesity, metabolic endotoxemia and dysbiosis: a randomized double-blind controlled clinical trial. Clin Nutr. 2014; 33(6): 973–981.
