Imam Journal of Applied Sciences

ORIGINAL ARTICLE
Year
: 2018  |  Volume : 3  |  Issue : 1  |  Page : 22--27

Changes in plasma alanine transaminase/serum glutamic-pyruvic transaminase, aspartate transaminase/serum glutamic-oxaloacetic transaminase, glutathione transferase, and albumin in rabbits given amoxicillin overdose supplemented with cucumber fruit juice


Mathew Folaranmi Olaniyan1, Oluwatoyin Ateni2,  
1 Department of Medical Laboratory Science, Edo University, Iyamho, Edo State, Nigeria
2 Department of Medical Laboratory Science, Achievers University, Owo, Ondo State, Nigeria

Correspondence Address:
Dr. Mathew Folaranmi Olaniyan
Department of Medical Laboratory Science, Edo University, Iyamho, Edo State
Nigeria

Abstract

Study Background: Raw Cucumber (Cucumis sativus) fruit juice contains substances of health-promoting bioactivities. Alanine transaminase/serum glutamic-pyruvic transaminase (ALT/SGPT), aspartate transaminase/serum glutamic-oxaloacetic transaminase (AST/SGOT), Glutathione S-Transferase (GST), and albumin are indices of hepatotoxicity, hepatitis, and liver damage which could be drug induced. Aim and Objective: This work was therefore designed to changes in plasma ALT/SGPT, AST/SGOT, GST, and albumin in rabbits given amoxicillin overdose supplemented with cucumber fruit juice. Materials and Methods: Fifteen rabbits of the same sex weighing 0.9–1.4 kg divided into three groups of 5 rabbits each were used for the study. Group A – 5 control rabbits; Group B – 5 rabbits given 30.0 mg/kgBW subcutaneous injection of amoxicillin every 24 h for 7 days which was followed by 30 ml raw cucumber fruit juice supplementation for 14 days; Group C – 5 rabbits given 30.0 mg/kgBW subcutaneous injection of amoxicillin every 24 h and raw cucumber fruit juice supplementation for 14 days simultaneously. ALT/SGPT, AST/SGOT, GST, and albumin were determined in the rabbits biochemically by spectrophotometry and fluorometry. Results: The results obtained showed a significant increase in plasma ALT, GST with a significant decrease in plasma albumin in the rabbits when the rabbits were given 30.0 mg/kgBW subcutaneous injection of amoxicillin compared with the results obtained from the control rabbits; their basal samples and when the rabbits were supplemented with 30 ml raw cucumber fruit (P < 0.05). There was a significant decrease in plasma ALT, AST, GST with a significant increase in plasma albumin in the rabbits when the rabbits were supplemented with 30 ml raw cucumber fruit juice for 14 days compared with the results obtained from the rabbits when the rabbits were given 30.0 mg/kgBW subcutaneous injection of amoxicillin (P < 0.05). Conclusion: This work revealed hepatoprotective raw cucumber fruit juice as there was no significant biochemical alterations when the rabbits were co-administered with raw cucumber fruit juice and amoxicillin overdose bioactivities while increase in plasma GST, AST, ALT, and decrease in albumin in amoxicillin overdose was reversed to normal plasma levels when the rabbits were given raw cucumber fruit juice. Raw cucumber fruit juice could be taken as antidote in hepatotoxicity.



How to cite this article:
Olaniyan MF, Ateni O. Changes in plasma alanine transaminase/serum glutamic-pyruvic transaminase, aspartate transaminase/serum glutamic-oxaloacetic transaminase, glutathione transferase, and albumin in rabbits given amoxicillin overdose supplemented with cucumber fruit juice.Imam J Appl Sci 2018;3:22-27


How to cite this URL:
Olaniyan MF, Ateni O. Changes in plasma alanine transaminase/serum glutamic-pyruvic transaminase, aspartate transaminase/serum glutamic-oxaloacetic transaminase, glutathione transferase, and albumin in rabbits given amoxicillin overdose supplemented with cucumber fruit juice. Imam J Appl Sci [serial online] 2018 [cited 2022 May 21 ];3:22-27
Available from: https://www.e-ijas.org/text.asp?2018/3/1/22/243624


Full Text



 Introduction



Cucumbers are a common fruit in Nigeria taken as beverage. The phytonutrients in raw cucumber fruit juice include calcium, carbohydrates, dietary fiber, folates, iron, magnesium, manganese, niacin, pantothenic acid, phosphorus, potassium, protein, pyridoxine, riboflavin, thiamin, Vitamin A, Vitamin C, Vitamin K, silica, and zinc. Its skin hydrated and naturally moist.[1],[2],[3],[4],[5],[6],[7],[8],[9]

Traditional applications of cucumber fruit extract include treatment of burns, edema, gout, high cholesterol, inflammation, intestinal parasites, pink eye, rheumatoid arthritis, sore throat, and ulcerative colitis and skin ailments (sunburn and under eye bags).[1],[2],[3] The water and fiber, cucumber contents aid digestion thereby preventing constipation. It is a natural diuretic the water in cucumber is of better quality than regular water, as it is naturally purified. Cucumber can prevent kidney stones, control blood pressure, and can lower cholesterol considering the constituent amounts of magnesium, potassium, and fiber.[4],[5] It is applied in the management of diabetes because with a rating of zero on the glycemic index, the carbohydrates in cucumbers can easily be digested by diabetics thereby serving as a source of nutrition to the patients during treatment for glucose homeostasis.[6],[7],[8]

Amoxicillin is one of the penicillin antibiotic drugs is used to treat many different types of infection caused by bacteria, such as tonsillitis, bronchitis, pneumonia, gonorrhea, and infections of the ear, nose, throat, skin, or urinary tract.[9],[10],[11],[12],[13] The amoxicillin overdose may be accidental and unintentional. Amoxicillin overdose could affect the kidneys, leading to poor kidney function and possibly kidney failure, bleeding, nausea, vomiting, and diarrhea.[14],[15],[16],[17],[18]

Alanine transaminase/serum glutamic-pyruvic transaminase (ALT/SGPT) and Aspartate transaminase/serum glutamic-oxaloacetic transaminase (AST/SGOT) are liver aminotransferases. The enzymes catalyze the interconversions of amino acids and α-oxoacids by transfer of amino groups.[19],[20],[21] Although AST is found in other cells and organs the level of AST and ALT increase in hepatitis and hepatocellular damage.[22],[23],[24],[25] AST is usually ordered together with ALT to diagnose liver damage/inflammation. AST/ALT ratio can be calculated to distinguish between different causes of liver damage. Hepatotoxicity may also increase the plasma levels of AST and ALT.[19],[20],[21]

Glutathione S-transferases (GSTs) are a group of, enzymes important in the detoxication of xenobiotics in mammals.[26],[27],[28],[29],[30],[31] GSTs protect cells against toxicants through biochemical conjugation of thiol group of the glutathione (GSH) to electrophilic xenobiotics, to defend cells against the mutagenic, carcinogenic, and toxic effects of the substances. GSTs act in plants, insects yeast, bacteria, in most mammalian tissues, especially in the liver, which plays a key role in detoxification.[26],[27],[28],[29],[30],[31]

Albumin is the most abundant of the serum proteins (albumin and globulin). It constitutes between 55% and 65% of the total protein. Albumin is produced in the liver, and the half-life is 2–3 weeks.[32],[33],[34],[35] The main biological functions of albumin are to maintain the water balance in serum and plasma and to transport and store a wide variety of ligands, for example, fatty acids, calcium, bilirubin, and hormones such as thyroxine. Low levels of albumin in the blood are associated with: impaired albumin synthesize in the liver; liver disease; kidney disease; malnutrition or malabsorption; generalized shock; inflammation; burns; dermatitis; intestinal disease; and Crohn's disease. High albumin levels in the blood have little clinical value except in dehydration.[32],[33],[34],[35]

Cucumber (Cucumis sativus) fruit is a common fruit contains phytochemicals and phytonutrients of health benefits. Amoxicillin is an antibiotic used to treat infections, and it is used by many people which could be overdosed causing hepatotoxicity. There is little information on the evaluation of plasma ALT/SGPT, AST/SGOT, GST and albumin in rabbits given amoxicillin overdose supplemented with raw cucumber fruit juice hence the need for this work.

This work was therefore designed to determine changes in plasma ALT/SGPT, AST/SGOT, GST and albumin in rabbits given amoxicillin overdose supplemented with raw cucumber fruit juice.

 Materials and Methods



Materials

Study area

This work was carried out at the Animal house of Achievers University, Owo-Nigeria. Achievers University is in Owo, Ondo State, Nigeria. The university is a private-sector initiative, established in 2007 and accredited by the National Universities Commission. It is located on land in the Idasen community of Owo, consisting of Ulale 1, Ulale 11, Ulema, Ijegunma, Isijogun, and Amurin Elegba (formerly Amurin, Ogain). The university sprang from the Achievers Group of Education and Training Organization, located in Ibadan Oyo State of Nigeria owned and run by Hon. Dr. Bode Ayorinde and other educationalists. The university commenced academic activities during the 2007/2008 academic session. In the Nigerian National University Commission annual university rankings for 2013, it was rated 53rd. It has three colleges which include college of natural and applied sciences; college of engineering and technology; and college of social science and management including a postgraduate school.

Study population

Fifteen rabbits of the same sex divided into three groups of five rabbits each was used for the study. The rabbits were bought animal farm in Owo-Nigeria and was presented to Federal School of Agriculture, Akure, for confirmation.

Group A – Five control rabbits were fed with normal meal and water throughout the investigationGroup B – Five rabbits given 30.0 mg/kgBW subcutaneous injection of amoxicillin every 24 h for 7 days which was followed by 30 ml raw cucumber fruit juice supplementation for 14 daysGroup C – Five rabbits given 30.0 mg/kgBW subcutaneous injection of amoxicillin every 24 h and raw cucumber fruit juice supplementation for 14 days simultaneously.

Administration of amoxicillin

Amoxicillin was bought from a registered pharmaceutical shop in Owo-Nigeria. Overdose of 30.0 mg/kgBW subcutaneous injection of amoxicillin was given every 24 h for 7 days.

Preparation of cucumber (Cucumis sativus) fruit juice

Cucumber (C. sativus) was bought from fruit vendors in Owo-Nigeria. The fruit was presented to Federal School of Agriculture, Akure for confirmation. The fruit was washed in sterile water and then sliced. The sliced fruit was blend together using an electronic blender. The raw fluid was extracted using a sterile sieve. The raw liquid extract was served to the rabbits as juice. 30 ml was given to the rabbits on daily basis. The juice will always be freshly prepared.

Specimen (blood) collection

Five milliliters of venous blood was collected from each of the rabbits into lithium heparinized bottles for the biochemical analysis.

Methods

Biochemical analysis

Plasma aspartate transaminase activities using Randox kit

Principle: This is an optimized standard method according to the concentrations recommended by the International Federation of Clinical Chemistry. α-oxoglutarate reacts with L-aspartate in the presence of AST to form L-glutamate plus oxaloacetate. The indicator reaction utilizes the oxaloacetate for a kinetic determination of nicotinamide-adenine dinucleotide (NADH) consumption.

Alanine aminotransferase activities using Randox kit

Principle: ALT catalyzes the transamination of L-alanine to α-ketoglutarate, forming L-glutamate and pyruvate. The pyruvate formed is reduced to lactate by lactate dehydrogenase with simultaneous oxidation of reduced NADH. The change in absorbance is directly proportional to the ALT activity and is measured using a bichromatic (340, 700 nm) rate technique.

Glutathione S-transferase by fluorescent activity using Detectx ® Kit of arbor assay

Principle: The DetectX ® GST Fluorescent Activity kit is designed to quantitatively measure the activity of GST present in a variety of samples. Please read the complete kit insert before performing this assay. A GST standard is provided to generate a standard curve for the assay, and all samples should be read off the standard curve. The kit utilizes a nonfluorescent molecule that is a substrate for the GST enzyme which covalently attaches to GSH to yield a highly fluorescent product. Mixing the sample or standard with the supplied detection reagent and GSH and incubating at room temperature for 30 min yields a fluorescent product which is read at 460 nm in a fluorescent plate reader with excitation at 390 nm.

Measurement of serum albumin using Randox kit

Principle: The measurement of serum albumin is based on its quantitative binding to the indicator 3, 3', 5, 5' tetrabromom cresol sulfonephthalein bromocresol green (BCG). The albumin BCG complex absorbs maximally at 578 nm, the absorbance being directly proportional to the concentration of albumin in the sample.

Method of data analysis

The information from this study was subjected to statistical analysis to determine mean, standard deviation, Students 't' and probability value at 0.05 level of significance using IBM SPSS 18.0 (Armonk, New York, United States).

 Results



There was a significant increase in plasma ALT, GST with a significant decrease in plasma albumin in the rabbits when the rabbits were given 30.0 mg/kgBW subcutaneous injection of amoxicillin every 24 h for 7 days compared with the results obtained from the control rabbits; their basal samples and when the rabbits were supplemented with 30 ml raw cucumber fruit juice for 14 days with P < 0.05 [Table 1], [Table 2] and [Figure 1].{Table 1}{Table 2}{Figure 1}

There was a significant decrease in plasma ALT, AST, GST with a significant increase in plasma albumin in the rabbits when the rabbits were supplemented with 30 ml raw cucumber fruit juice for 14 days compared with the results obtained from the rabbits when the rabbits were given 30.0 mg/kgBW subcutaneous injection of amoxicillin every 24 h for 7 days with P < 0.05 [Table 1], [Table 2] and [Figure 1].

There was no significant difference in the plasma values of plasma ALT, GST, albumin, and AST when the rabbits were given 30.0 mg/kgBW subcutaneous injection of amoxicillin every 24 h and raw cucumber fruit juice supplementation for 14 days simultaneously with P > 0.05 [Table 1], [Table 2] and [Figure 1].

No significant difference was also obtained in the plasma AST when the rabbits were given 30.0 mg/kgBW subcutaneous injection of amoxicillin every 24 h for 7 days compared with the results obtained from the control rabbits and their basal samples P > 0.05 [Table 1], [Table 2] and [Figure 1].

 Discussion



There was a significant increase in plasma ALT, GST with a significant decrease in plasma albumin in the rabbits when the rabbits were given 30.0 mg/kgBW subcutaneous injection of amoxicillin every 24 h for 7 days compared with the results obtained from the control rabbits; their basal samples and when the rabbits were supplemented with 30 ml raw cucumber fruit juice for 14 days.

Increase in plasma ALT, GST with decrease in plasma albumin could be attributed to amoxicillin drug-induced hepatotoxicity as high intracellular concentrations of GSTs coupled with their cell-specific cellular distribution allows them to function as biomarkers for localizing and monitoring injury to defined cell types.[36],[37],[38] For instance, liver cells are very rich in alpha GST which its serum level increases in hepatocellular injury, transplantation, toxicity, and viral infections.[36],[37],[38] GST polymorphisms of the alpha, mu, pi, and theta classes are implicated in asthma, atherosclerosis, allergies, and other inflammatory diseases.[36],[37],[38] In addition, hypoalbuminemia and increased plasma ALT can be caused by liver damage, hepatitis, and hepatotoxicity because liver synthesizes albumin while ALT is majorly found in the liver.[19],[21],[25] Serum ALT level and AST are commonly measured to determine hepatitis and hepatocellular damage possibly due to toxic substances such as drug and infectious agents.[33] ALT is commonly measured clinically as a part of a diagnostic evaluation of hepatocellular injury and to determine liver health.[24],[35]

There was a significant decrease in plasma ALT, AST, GST with a significant increase in plasma Albumin in the rabbits when the rabbits were supplemented with 30 ml raw cucumber fruit juice for 14 days compared with the results obtained from the rabbits when the rabbits were given 30.0 mg/kgBW subcutaneous injection of amoxicillin every 24 h for 7 days. This could be attributed to health-promoting bioactivities of raw cucumber fruit juice as a large percentage of this fruit is water which makes it an ideal drink for a healthier, cleaner, and toxin-free body. Cucumber juice can be used to treat dehydration due to its high water content and electrolyte-restoring properties. The juice can eliminate those free radicals before they can do serious damage.[39],[40]

Notable levels of Vitamin C in this juice make it a great choice for boosting the immune system. It can stimulate the production of white blood cells, which are the body's 1st line of defense and also behave as an antioxidant, seeking out, and neutralizing harmful-free radicals. People associate calcium with bone health, but the level of calcium in the blood is also a key determinant of the hormonal balance. If the thyroid gland or pituitary gland begins to malfunction, high levels of calcium from this juice can help to keep the hormone levels normal.[39],[40] This could also be attributed to the phytonutrients constituents in raw cucumber fruit juice which include: calcium, carbohydrates, dietary fiber, folates, iron, magnesium, manganese, niacin, pantothenic acid, phosphorus, potassium, protein, pyridoxine, riboflavin, thiamin, Vitamin A, Vitamin C, Vitamin K, and zinc.[1],[2],[3],[4],[5],[6],[7],[8] As many of the phytonutrients (pyridoxine, riboflavin, thiamin, Vitamin A, Vitamin C, Vitamin K, and zinc) of them have antioxidative bioactivities.[8]

 Conclusion



This work revealed hepatoprotective effect raw cucumber fruit juice as there was no significant biochemical alterations when the rabbits were coadministered with raw cucumber fruit juice and amoxicillin overdose bioactivities while increase in plasma GST, AST, ALT, and decrease in albumin in amoxicillin overdose was reversed to normal plasma levels when the rabbits were given raw cucumber fruit juice.

Recommendation

Raw cucumber fruit juice could be taken as antidote in hepatotoxicity.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

References

1Schieberle P, Ofner S, Grosch W. Evaluation of potent odorants in cucumbers (Cucumis sativus) and muskmelons (Cucumis melo) by aroma extract dilution analysis. J Food Sci 1990;55:193.
2Peter JJ, Nick T, Thorpe IJ. Ancient Inventions Chapter 12 Sport and Leusure: Roman Gardening Technology. Ballantine Books; New York: Ballantine Books, 1995. p. 563.
3Kuhnlein HV, Turner NJ. Traditional Plant Foods of Canadian Indigenous Peoples: Nutrition, Botany and Use. Amsterdam, Netherlands: Gordon and Breach; 1996. p. 159.
4Robert WD. From work on the petra papyri: Arabic on a Greek Ostracon from Roman Egypt and the name of the church father Sozomen (PDF). Zeitschrift für Papyrologie und Epigraphik 2000;131:173-6.
5Renner SS, Schaefer H, Kocyan A. Phylogenetics of cucumis (Cucurbitaceae): Cucumber (C. Sativus) belongs in an Asian/Australian clade far from melon (C. Melo). BMC Evol Biol 2007;7:58.
6James G. E. coli Outbreak is a New Strain. BBC News; 2011.
7David B. Taste, Memory: Forgotten Foods, Lost Flavors, and why They Matter. VT, USA: Chelsea Green Publishing; 2012. p. 109.
8Shang Y, Ma Y, Zhou Y, Zhang H, Duan L, Chen H, et al. Plant science. Biosynthesis, regulation, and domestication of bitterness in cucumber. Science 2014;346:1084-8.
9Janos F, Ganellin C. Robin. Analogue-based Drug Discovery. John Wiley and Sons; 2006. p. 490.
10Deirdre K. Diseases of the Liver and Biliary System in Children. 3rd ed. Chichester, UK: Wiley-Blackwell; 2008. p. 217.
11Jiben R. An Introduction to Pharmaceutical Sciences Production, Chemistry, Techniques and Technology. Cambridge: Woodhead Pub; 2012. p. 239.
12Ahovuo-Saloranta A, Rautakorpi UM, Borisenko OV, Liira H, Williams JW. Mäkelä M Antibiotics for acute maxillary sinusitis in adults. Cochrane Database Syst Rev 2014;(2):CD000243.
13Gillies M, Ranakusuma A, Hoffmann T, Thorning S, McGuire T, Glasziou P, et al. Common harms from amoxicillin: A systematic review and meta-analysis of randomized placebo-controlled trials for any indication. CMAJ 2015;187:E21-31.
14Kagan BM. Ampicillin rash. West J Med 1977;126:333-5.
15Pichichero ME. A review of evidence supporting the American academy of pediatrics recommendation for prescribing cephalosporin antibiotics for penicillin-allergic patients. Pediatrics 2005;115:1048-57.
16Barton DS. The Child's Health: The Parents' One-stop Reference Guide to Symptoms, Emergencies, Common Illnesses, Behavior Problems, Healthy Development. 2nd ed. New York: Bantam Books; 2005.
17Cundiff J, Joe S. Amoxicillin-clavulanic acid-induced hepatitis. Am J Otolaryngol 2007;28:28-30.
18Baselt R. Disposition of Toxic Drugs and Chemicals in Man. 8th ed. Foster City, CA: Biomedical Publications; 2008. p. 81-3.
19Kirsch JF, Eichele G, Ford GC, Vincent MG, Jansonius JN, Gehring H, et al. Mechanism of action of aspartate aminotransferase proposed on the basis of its spatial structure. J Mol Biol 1984;174:497-525.
20Hayashi H, Wada H, Yoshimura T, Esaki N, Soda K. Recent topics in pyridoxal 5'-phosphate enzyme studies. Annu Rev Biochem 1990;59:87-110.
21Berg JM, Tymoczko JL, Stryer L. Biochemistry. 6th Edition. New York: W.H. Freeman; New York: 2006. p. 656-60.
22Watkins PB, Kaplowitz N, Slattery JT, Colonese CR, Colucci SV, Stewart PW, et al. Aminotransferase elevations in healthy adults receiving 4 grams of acetaminophen daily: A randomized controlled trial. JAMA 2006;296:87-93.
23Koski RR. Omega-3-acid ethyl esters (Lovaza) for severe hypertriglyceridemia. Pharm Ther 2008;33:271-303.
24Ghouri N, Preiss D, Sattar N. Liver enzymes, nonalcoholic fatty liver disease, and incident cardiovascular disease: A narrative review and clinical perspective of prospective data. Hepatology 2010;52:1156-61.
25Wang CS, Chang TT, Yao WJ, Wang ST, Chou P. Impact of increasing alanine aminotransferase levels within normal range on incident diabetes. J Formos Med Assoc 2012;111:201-8.
26Douglas KT. Mechanism of action of glutathione-dependent enzymes. Adv Enzymol Relat Areas Mol Biol 1987;59:103-67.
27Boyer TD. The glutathione S-transferases: An update. Hepatology 1989;9:486-96.
28Sheehan D, Meade G, Foley VM, Dowd CA. Structure, function and evolution of glutathione transferases: Implications for classification of non-mammalian members of an ancient enzyme superfamily. Biochem J 2001;360:1-6.
29Atkinson HJ, Babbitt PC. Glutathione transferases are structural and functional outliers in the thioredoxin fold. Biochemistry 2009;48:11108-16.
30Mukanganyama S, Bezabih M, Robert M, Ngadjui BT, Kapche GF, Ngandeu F, et al. The evaluation of novel natural products as inhibitors of human glutathione transferase P1-1. J Enzyme Inhib Med Chem 2011;26:460-7.
31Oakley A. Glutathione transferases: A structural perspective. Drug Metab Rev 2011;43:138-51.
32Kenneth H, Dallas HW, Schlossberg J, Hurst W. In: Boyter L, Charles H, editors. Clinical Methods: The History, Physical, and Laboratory Examinations. 3rd ed. Boston: Butterworths; 1990. p. 101.
33Douglas MA. Dorland's Illustrated Medical Dictionary. 29th ed. Philadelphia, PA: Saunders; 2000. p. 860.
34Green P, Woglom AE, Genereux P, Daneault B, Paradis JM, Schnell S, et al. The impact of frailty status on survival after transcatheter aortic valve replacement in older adults with severe aortic stenosis: A single-center experience. JACC Cardiovasc Interv 2012;5:974-81.
35Mutlu EA, Keshavarzian A, Mutlu GM. Hyperalbuminemia and elevated transaminases associated with high-protein diet. Scand J Gastroenterol 2006;41:759-60.
36Beckett GJ, Chapman BJ, Dyson EH, Hayes JD. Plasma glutathione S-transferase measurements after paracetamol overdose: Evidence for early hepatocellular damage. Gut 1985;26:26-31.
37Loguercio C, Caporaso N, Tuccillo C, Morisco F, Del Vecchio Blanco G, Del Vecchio Blanco C, et al. Alpha-glutathione transferases in HCV-related chronic hepatitis: A new predictive index of response to interferon therapy? J Hepatol 1998;28:390-5.
38Franco R, Schoneveld OJ, Pappa A, Panayiotidis MI. The central role of glutathione in the pathophysiology of human diseases. Arch Physiol Biochem 2007;113:234-58.
39Nonnecke IL. Vegetable Production. Springer USA; Springer-Verlag US: 1st edition. 1989. p. 658.
40Huang S, Li R, Zhang Z, Li L, Gu X, Fan W, et al. The genome of the cucumber, Cucumis sativus L. Nat Genet 2009;41:1275-81.