1 Türk Biyokimya Dergisi [Turkish Journal of Biochemistry Turk J Biochem] 2015; 40(1):66 73 doi: /tjb Research Article [Araştırma Makalesi] Yayın tarihi 24 Ocak 2015 TurkJBiochem.com [Published online January 24, 2015] The effects of grape seed extract against toxicity of benzene on liver and kidney tissues of albino mice: biochemical evaluation [Albino farelerde benzenin böbrek ve karaciğer dokularında toksisitesine karşı üzüm çekirdeği ekstraktının etkisi: biyokimyasal değerlendirme] Kültiğin Çavuşoğlu 1, Emine Yalçın 1, Kürşad Yapar 2, Ertan Oruç 3, Birgül Gür 1, Figen Çiçek 4 1 Giresun University, Faculty of Science and Arts, Department of Biology, Biology Department, Giresun 2 Giresun University, Faculty of Medicine, Department of Medical Pharmacology, Giresun 3 Ataturk University, Faculty of Veterinary Medicine, Department of Pathology, Erzurum 4 Giresun University, Vocational School of Health Services, Department of Medical Services and Techniques, Giresun ABSTRACT Objective: The objective of the present study was to investigate the possible effects of grape seed extract (GSE) against benzene-induced toxicity in Swiss albino mice. Methods: The animals were randomly divided into six groups each containing six mice. Group I, treated with distilled water; Group II and III orally treated with 50 mg/kg and 150 mg/kg body weight GSE, respectively. Group IV, orally treated with 250 mg/kg body weight benzene by using feeding cannula; Group V, orally treated with 50 mg/kg body weight GSE mg/kg body weight benzene; Group VI, orally treated with 150 mg/kg body weight GSE mg/kg of body weight benzene for 50 consecutive days. At the end of experimental period all mice were sacrificed; blood, liver and kidney tissues were removed after post-mortem examination. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), blood urea nitrogen (BUN), creatinine levels were analyzed from serum. Malondialdehyde (MDA) and reduced glutathione (GSH) levels were analyzed from isolated tissues. And also histopathological examinations of liver and kidney tissues were investigated. Results: Serum AST, ALT, ALP, BUN and creatinine levels were slightly increased in Group IV compared with the other tested groups (p<.05). Benzene-induced toxicity caused a significant decrease in GSH levels and a significant rise in MDA levels of liver and kidney tissues. Oral treatment with GSE significantly ameliorated the indices of hepatotoxicity, nephrotoxicity and lipid peroxidation induced by benzene. Both doses of GSE provided significant protection and the strongest effects were observed at the dose level of 150 mg/kg. Conclusion: Consequently, it was found that GSE has a significant positive effect in benzeneinduced toxicity, and its GSE effect is dose dependent. Key Words: Benzene, GSH, MDA grape seed Conflict of Interest: The authors have no conflict of interest. Correspondence Address [Yazışma Adresi] Kültiğin Çavuşoğlu, MD. Giresun Üniversitesi, Fen Edebiyat Fakültesi, Biyoloji Bölümü, Türkiye Phone: Registered: 28 June 2013; Accepted: 25 September 2014 [Kayıt Tarihi: 28 Haziran 2013; Kabul Tarihi: 25 Eylül 2014] ÖZET Amaç: Bu çalışmanın amacı albino farelerde benzen tarafından oluşturulan toksisitesiye karşı üzüm çekirdeği ekstraktının muhtemel etkisini araştırmaktır. Metod: Her bir grupta 6 fare olmak üzere uygulama grupları oluşturulmuştur. Grup I, oral yolla distile su ile; Grup II, oral yolla 50 mg/kg GSE; Grup III, 150 mg/kg GSE; Grup IV, oral yolla kanula kullanılarak 250 mg/kg benzen; Grup V, 50 mg/kg GSE mg/kg benzen; Grup VI, 150 mg/kg GSE mg/kg benzen ile 50 gün süre ile beslenmişlerdir. Deney sürelerinin sonunda otopsi yoluyla farelerin kan, karaciğer ve böbrek dokuları alınmıştır. Aspartat aminotransferaz (AST), Alanin aminotransferaz (ALT), Alkalen fosfataz (ALP), kan üre azotu (BUN), kreatinin düzeyleri kan dokudan, malondialdehit (MDA), redükte glutatyon (GSH) ise karaciğer ve böbrek dokularında incelenmiştir. Ayrıca karaciğer ve böbrek dokularında histopatolojik değişimler de araştırılmıştır. Bulgular: Benzen uygulanan farelerde serum AST, ALT, ALP BUN ve kreatinin seviyelerinde kontrol grubuna kıyasla düşük düzeyde artış belirlenmiştir. Benzen tarafından oluşturulan oksidatif toksisitenin karaciğer ve böbrek dokularında GSH seviyesinde önemli bir azalmaya ve MDA seviyesinde önemli bir artışa neden olduğu belirlenmiştir. Üzüm çekirdeği ekstraktı uygulamasının benzen tarafından oluşturulan hepatotoksisite, nefrotoksisite ve lipid peroksidasyonunda önemli gerilemeye neden olduğu belirlenmiştir. Üzüm çekirdeği ekstraktının test edilen iki dozunun benzen tarafından oluşturulan toksisite üzerine pozitif etkisi olduğu belirlenmiş ve en güçlü etkiyi 150 mg/kg dozunda gösterdiği belirlenmiştir. Sonuç: Sonuç olarak GSE nin benzen toksisitesine karşı pozitif etkisi olduğu ve bu etkinin doza bağımlı olduğu belirlenmiştir. Anahtar Kelimeler: Benzen, GSH, MDA, üzüm çekirdeği ekstraktı Çıkar Çatışması: Yazarların çıkar çatışması yoktur ISSN X (electronic) (printed)
2 Introduction Benzene is an important pollutant compound, present in both occupational and general environment. It is a highly flammable liquid with clastogenic and carcinogenic activity [1-3]. It is classified as a Known carcinogen Category A under the risk assessment Guidelines of 1986 . Many industrial applications have been found for benzene, and since its discovery it has been widely used as an industrial solvent . Potential exposure to benzene can be higher in certain industries, such as shoe factories, petrol stations and the petrochemical industry . Furthermore, lifestyle factors, such as cigarette smoking is the main source of benzene exposure for many people . Humans with acute inhalation exposure to benzene may experience drowsiness, dizziness, headaches and a host of eye, skin, and respiratory tract irritations [3-5]. Many experimental animal studies, support the evidence that benzene exposure increases the risk of cancer in multiple organ systems including the hematopoietic system, liver and stomach . Acute occupational exposure to benzene may cause narcosis: headache, dizziness, drowsiness, confusion, tremors and loss of consciousness. Use of alcohol enhances the toxic effect. Chronic exposure to benzene can reduce the production of both red and white blood cells from bone marrow in humans, resulting in aplastic anaemia [5,6]. Benzene is metabolized, primarily in the liver, to a variety of hydroxylated and ring-opened products that are transported to the periferal tissues where subsequent secondary metabolism occurs. Benzene and its metabolites may damage cellular macromolecules to induce toxicity include the covalent binding of reactive metabolites of benzene . Decreasing the hepatic metabolism of benzene by partial hepatectomy also reduced benzene toxicity, suggesting that hepatic metabolism plays an important role in toxicity. In addition to hepatic metabolism, it appears that filtration of benzene and its metabolites contributes to toxicity [8,9]. So in this study the toxic effects of benzene on liver and kidney tissues also investigated. It was known that exposure to benzene causes oxidative stress and damage . MDA one of the known secondary products of lipid peroxidation induced by oxidative damage, may be used as an indicator of cell membrane injury . Due to its large role in countering increases in lipid peroxidation which occurs during oxidative stress, the response of GSH has been frequently studied. GSH is an important antioxidant, preventing damage to cellular components caused by oxidative stress . So we investigated the MDA and GSH levels and tissue pathology for determine oxidative damage of benzene on liver and kidney tissues. The use of certain materials may help to decrease the toxicity created by benzene. Recently, biopolymer materials such as zinc, selenium and leaf extract of Ocimum basilicum L. have been used for decreasing the toxic effects of benzene [13,14]. Grape seed extract (GSE) is a natural extract obtained from the seed of grape . Grapes and grape products are good sources of dietary flavonoids, which are powerful antioxidant compounds . GSE is a complex mixture of polyphenols containing dimers, trimers, and other oligomers (procyanidins) of catechin and epicatechin . Several experimental studies have demonstrated that GSE is highly bioavailable and provides significantly greater protection against free radicals . In order to overcome the potential harmful effect of benzene, GSE was tried as a toxicity-limited agent. For determine the effects of GSE, different GSE doses were orally given to the animals through benzene treatment. The aim of this study is to investigate the toxicity of benzene on liver and kidney tissues specifically out of MDA and GSH level measurements and the beneficial effects of GSE against benzene induced toxicity in an animal model. Recently, genomic data has been added to techniques to make close comparisons between species and determine relatedness. Humans share about 90% of genome with mice . Scientists have been able to take advantage of these similarities in generating experimental and predictive models of human disease. So in this study the toxicity of benzene was investigated in an animal-mice model. Materials and Methods Animals and GSE GSE (formerly Grape Seed PCO Phytosome tabs) was purchased from Health Genesis Corp. (Bay Harbor Island, FL, USA). A total number of 36 adult male Mus musculus var. albino mice weighing g were used in the current study. Healthy mice were obtained from the Animal Research Center of the Refik Saydam Hifzissiha Institute (Ankara, Turkey). All animals were housed in 26x15x50 cm stainless steel cages and kept under controlled laboratory conditions of 22±3 o C and 55±5% relative humidity with a 12-hour light dark cycle throughout the experiment. The animals were allowed to acclimatize for 1 week before the planned experimental test and fed a standard pellet diet (Samsun Food Industry, Samsun, Turkey). In this study, the methods and techniques applied to mice were carried out according to the guidelines set by the World Health Organization (Geneva, Switzerland) and the ethical standards of the local ethical committee for animal experiments at Giresun University (B.30.2.G RE / ). Experimental protocol The animals were divided into six groups each containing six mice. Distilled water was administered to mice in Group I and served as control . Mice in Group II and III were fed orally with two different doses of GSE as 50 mg/kg and 150 mg/kg of body weight, respectively. Group IV was fed orally by using feeding cannula with 250 mg/kg of body weight benzene; Group V was treated with 50 mg/kg of body weight GSE mg/kg of body Turk J Biochem 2015; 40(1): Çavuşoğlu et al.
3 weight benzene. Group VI was treated with 150 mg/kg of body weight GSE mg/kg of body weight benzene. The dose of benzene used in this study was determined to be 250 mg/kg. This dose was chosen because it induced an increase in the frequency of toxicity that was essential to determine the beneficial role of GSE [21,22]. In this study the experimental period was applied for 50 days to determine the long effect of benzene in accordance with literature. In some studies the toxicity of benzene was investigated for 3 days  or 6 days  for determine the short term effect; also in some studies the toxicity was studied for 30 days  or 50 days  to determine the long term effect. GSE doses were chosen that were comparable to those daily consumption amounts recommended by practitioners of nutritional medicine to support optimal health, and 50 and 150 mg/kg of body weight were effective doses for protection by GSE . Serum analysis For serum isolation, blood samples were collected by cardiac puncture with the animal under mild ether anesthesia. Blood samples were drawn directly into precooled vacutainer tubes (BD Vacutainer Systems, San Jose, CA, USA) for storage and centrifuged at g for 10 minutes at 4 o C. Aspartate aminotransferase (AST) (AST/ GOT liquid reagent, catalog number A , Teco Diagnostics, Anaheim, CA, USA), alanine aminotransferase (ALT) (ALT/GPT liquid reagent, catalog number A , Teco Diagnostics) and alkaline phosphatase (ALP) (Liquid kinetic, catalog number A , Teco Diagnostics) enzyme activities, blood urea nitrogen (BUN) (catalog number B , Teco Diagnostics) and creatinine (catalog number C , Teco Diagnostics) concentrations were measured by commercially available kits using an autoanalyzer (model 99M Chemistry Analyzer, Medispec, Germantown, MD, USA). MDA and GSH levels At the end of the experimental period, the mice were fasted overnight sacrificed by cardiac exsanguination under ether anesthesia. For histopathology investigation liver and kidney tissues were carefully collected from control and experimental groups at the end of 50 th day. The tissues were immediately washed, dried, and processed for biochemical measurements. Sample tissues were then homogenized in 0.15 M ice-cold KCl for 3 minutes at rpm with a homogenizer (Ultraturrax type T25-B, IKA Labortechnik, Staufen, Germany). Homogenates were centrifuged at g at 4 o C for 1 hour. The supernatants were collected and stored at 40 o C until used for analyses . GSH and MDA levels in related tissues were measured spectrophotometrically (UV mini-1240, Shimadzu, Kyoto, Japan) using the colorimetric methods described by Beutler et al.  and of Yoshoiko et al. , respectively. Histopathological examinations For light microscopic examination, fresh tissue samples including the liver and kidneys were fixed in 10% neutral Table 1. Effect of grape seed extract administration on selected biochemical parameters in albino mice treated with benzene Parameter Group I Group II Group III Group IV Group V Group VI Statistical significance (Control) (GSE 50) (GSE 150) (Benzene) (GSE 50+Benzene) (GSE150+Benzene) AST (U/L) ±7.28 d ±980 d ±11.23 d ±8.09 a ±11.73 b ±13.53 c d: p>0.05; a,b,c: <0.05 ALT (U/L) 46.53±4.42 c 46.98±2.93 c 48.33±3.71 c 66.17±5.42 a 59.50±5.52 b 51.16±6.60 c c: p>0.05; a,b: <0.05 ALP (U/L) ±8.04 c ±12.25 c ±6.13 c ±5.45 a ±7.72 b ±7.64 c c: p>0.05; a,b: <0.05 BUN (mg/l) ±7.67 d ±9.63 d ±6.06 d ±9.67 a ±10.84 b ±9.59 c d: p>0.05; a,b,c: <0.05 Creatinine (mg/l) 5.50±0.46 d 5.50±0.51 d 5.42±0.40 d 9.69±0.66 a 8.34±0.67 b 7.06±0.65 c d: p>0.05; a,b,c: <0.05 * Data are mean±sd values (n=6). Statistical significance between means was assessed using one-way analysis of variance followed by Duncan s test as a post-analysis of variance test and all variables were normally distributed. Within each line, means not significantly different share the same letter; means with no common letters are significantly different from each other. Turk J Biochem 2015; 40(1): Çavuşoğlu et al.
4 buffered formalin solution for routine processing, embedded in paraffin wax, sectioned at 5 µm, and stained with hematoxylin and eosin (H-E). Histopathological changes were semiquantitatively assessed under the light microscope . Statistical analysis The statistical analysis software SPSS for Windows version 10.0 (SPSS, Inc., Chicago, IL, USA) was used for statistical data analysis. Statistically significant differences between the groups were compared using one-way analysis of variance and Duncan s test. The data are given as mean ±SD values, and values of P<.05 are considered statistically significant. Results Serum enzyme parameters Table 1 demonstrates the changes in serum AST, ALT, ALP, BUN, and creatinine levels of all treatment groups. There were no significant differences in the levels of AST, ALT, ALP, BUN and creatinine among the control group and the groups treated with GSE alone (p>.05). Serum AST, ALT, ALP, BUN, and creatinine levels significantly increased after oral benzene exposure (p<.05). Oral administration of the two different doses of GSE decreased the enzyme activities in Groups V and VI compared with Group IV, but values were still significantly higher compared to control group. Also, GSE had an effect on BUN and creatinine levels, which are sensitive markers of kidney injure. Serum levels of BUN and creatinine significantly decreased (p<.05) in Group V and VI compared to group treated with benzene alone. In Group VI, the mean level of BUN was about 1.09-fold lower and the mean level of creatinine was about 1.37-fold lower than in Group IV. MDA and GSH levels Table 2 shows the alterations of GSH and MDA levels in liver and kidney tissues. No significant differences were observed among the control group, Group II and III in MDA and GSH levels of the liver and kidney tissues (p>.05). GSH levels were significantly decreased, whereas MDA levels were increased in liver and kidney tissues in Group IV, V and VI compared to Group I (p<.05). However, oral administration of GSE with two doses in Group V and VI was reversed the GSH and MDA levels back to the control levels in the liver and kidney tissues (p<.05). Pathological findings Similar liver and kidney histology were observed in control group and group III (Fig. 1,2). Severe histopathological changes were observed in group IV and V. In these groups, hyperemia in the central veins and sinusoids, degenerative and necrotic hepatocytes in liver; hyperemia and haemorrhagia, degeneration of tubular epithelium and intratubulary haemorrhagia in kidney were detected (Fig. 3-5). In the group VI, similar changes were observed, but Table 2. Effect of grape seed extract administration on malondialdehyde and reduced glutathione in liver and kidneys of albino mice treated with benzene Tissue parameter Group I Group II Group III Group IV Group V Group VI Statistical significance (Control) (GSE 50) (GSE 150) (Benzene) (GSE 50+Benzene) (GSE150+Benzene) MDA Liver (nmol/g) 0.26±0.03 d 0.26±0.02 d 0.26±0.03 d 0.82±0.07 a 0.58±0.06 b 0.43±0.02 c d: p>0.05; a,b,c: <0.05 MDA Kidney (nmol/g) 0.28±0.01 d 0.28±0.02 d 0.28±0.02 d 0.88±0.03 a 0.70±0.02 b 0.39±0.02 c d: p>0.05; a,b,c: <0.05 GSH Liver (mg/g) 0.55±0.04 a 0.53±0.03 a 0.54±0.04 a 0.17±0.02 d 0.26±0.03 c 0.37±0.05 b a: p>0.05; b,c,d: <0.05 GSH Kidney (mg/g) 0.61±0.02 a 0.61±0.02 a 0.61±0.03 a 0.20±0.02 d 0.25±0.03 c 0.36±0.02 b a: p>0.05; b,c,d: <0.05 *Data are mean±sd values (n=6). Statistical significance between means was assessed using one-way analysis of variance followed by Duncan s test as a post-analysis of variance test and all variables were normally distributed. Within each line, means not significantly different share the same letter; means with no common letters are significantly different from each other. Turk J Biochem 2015; 40(1): Çavuşoğlu et al.
5 (a) Figure 1. Group I (control group). The histological appearance of the liver (a) and kidney is normal. Hematoxylin and Eosin. (a) Figure 2. Group III (GSE 150). The hepatocytes (a) and tubules of the liver and kidney revealed normal histological architecture. Hematoxylin and Eosin. (a) Figure 3. Group IV (Benzen). The liver showed degenerative or necrotic alterations of hepatocytes (arrows) (a); the kidneys contained haemorrhagia foci (black arrows) and pyknotic nuclei of tubulary epithelium. Hematoxylin and Eosin. the severity of intratubular haemorrhagia was less frequently than group IV and V (Fig. 5). Discussion In this study the toxic effects of benzene on biochemical parameters of albino mice was investigated. AST, ALT and ALP levels of benzene treated group were increased compared to control group. This increase may be mainly due to the leakage of these enzymes from the liver cytosol into the blood stream . Lipid soluble benzene is transported in the blood and absorbed by different cell membranes. It tends to accumulate in tissues and about 50% of the absorbed dose may be eliminated unchanged while the remaining is metabolized in liver . In this Turk J Biochem 2015; 40(1): Çavuşoğlu et al.
6 (a) Figure 4. Group V (GSE 50+Benzen). The liver showed necrotic alterations of hepatocytes (arrows) (a) The kidneys contained haemorrhagia foci (black arrow). Hematoxylin and Eosin. (a) Figure 5. Group VI (GSE 150+Benzen). Hepatocytes had pyknotic nuclei (arrows) (a) and there was intratubular hyaline accumulation (black arrows) in the kidneys, but there was no haemorrhagia. Hematoxylin and Eosin. pathway liver cell membranes were selectively damaged by benzene although to different extents. When liver cells are damaged, AST, ALT and ALP enzymes released into the bloodstream [34,35] and the levels of these enzymes were increased. BUN and creatinine are nitrogenous waste products that are eliminated by kidneys, when excretion is suppressed in renal insufficiency . The effect of benzene on BUN and creatinine levels has also been described by other authors [4,25]. BUN and creatinine levels in benzene treated groups were higher than in control groups. Administration of two different doses of GSE significantly decreased the levels of AST, ALT, ALP, BUN, and creatinine in Groups V and VI compared with Group IV. Similarly, Yalcin et al.  reported that GSE treatment reversed the doxorubicin-induced increase in serum AST, ALT, BUN, and creatinine levels in albino mice. It was concluded that benzene administration resulted a significant increase in serum creatinine and BUN indicating significant damage to the kidney accompanied by degeneration of tubular epithelium and intratubulary haemorrhagia in kidney. The pathological changes caused kidney failure were verified from histopathology test results. Benzene and its metabolites are known to produce oxidized species and reactive oxygen indicating the increased risk of cell membrane damage . MDA is one of the most known secondary products of lipid peroxidation, and it can be used as a marker of cell membrane injury . In present study, we found that the application of benzene led to a significant increase in MDA levels, which is in agreement with similar studies [39,40]. GSH and its metabolizing enzymes provide the major antioxidant defense against reactive oxygen species induced cellular damage [41,42]. In contrast to MDA, GSH levels profoundly decreased in kidney and liver tissues. Benzene and its metabolites are known to produce oxidized species indicating the increased risk of cell membrane damage . The decrease in GSH level and increase in MDA level were associated with production of radicals and increased lipid peroxidation. GSE treatment in Group V and VI had a beneficial role in benzene toxicity and the levels of MDA and GSH were neared to control levels in these groups. The potent protective effect of GSE as antioxidant in suppressing lipid peroxidation was reported before [43,44]. GSE showed potent antioxidant activity by inhibiting free radical formation, delaying lipid oxidation, and reducing the concentration of H 2 O 2 produced by the oxidative stress . GSE is effective in preventing Turk J Biochem 2015; 40(1): Çavuşoğlu et al.
7 the oxidative stress associated loss of membrane surface charge which maintains the membrane integrity and function . As a result, it is concluded that, the findings of the present study demonstrated the beneficial role of GSE against benzene toxicity via its antioxidant capacity. Therefore, the antioxidant role of GSE may be used as a toxicity-limiting agent to reduce effects on human health of chemical agents in the near future. Conflict of Interest There are no conflicts of interest among the authors. References  Zhang L, Eastmond DA, Smith MT. The nature of chromosomal aberrations detected in humans exposed to benzene. Crit Rev Toxicol 2002; 32(1):1-42.  Holecková B, Piesová E, Sivikova K, Dianovskỳ J. Chromosomal aberrations in humans induced by benzene. Ann Agric Environ Med 2004; 11(2):  Dere E, Gyborova S, Aydın H. The effect of benzene on serum hormones and the activity of some enzymes in different tissues of rats. Acta Veterinaria 2003; 53:  Abousalem M, Elgerwi A, El-Mashad A. Biotoxic and haemotoxic effects of air pollutants at a benzene station on albino rat. Int J Anim Vet Adv 2012; 4:  IPCS (1993). Benzene. Geneva, World Health Organization, International Programme on Chemical Safety (Environmental Health Criteria 150; ehc150.htm).  Baan R, Grosse Y, Straif K, Secretan B, El Ghissassi F, et al. A review of human carcinogens--part F: chemical agents and related occupations. Lancet Oncol 2009; 10(12):  Snyder R, Hedli CC. An overview of benzene metabolism. Environ Health Perspect 1996; 104 Suppl 6:  Andrews LS, Sasame HA, Gillette JR. 3H-Benzene metabolism in rabbit bone marrow. Life Sci 1979; 25(7):  Irons RD, Dent JG, Baker TS, Rickert DE. Benzene is metabolized and covalently bound in bone marrow in situ. Chem Biol Interact 1980; 30(2):  Uzma N, Kumar BS, Hazari MA. Exposure to benzene induces oxidative stress, alters the immune response and expression of p53 in gasoline filling workers. Am J Ind Med 2010; 53(12):  Santos MT, Valles J, Aznar J, Vilches J. Determination of plasma malondialdehyde-like material and its clinical application in stroke patients. J Clin Pathol 1980; 33(10):  Pompella A, Visvikis A, Paolicchi A, De Tata V, Casini AF. The changing faces of glutathione, a cellular protagonist. Biochem Pharmacol 2003; 66(8):  Ibrahim KS, Saleh ZA, Farrag AR, Shaban EE. Protective effects of zinc and selenium against benzene toxicity in rats. Toxicol Ind Health 2011; 27(6):  Saha S, Mukhopadhyay MK, Ghosh PD, Nath D. Effect of Methanolic Leaf Extract of Ocimum basilicum L. on Benzene-Induced Hematotoxicity in Mice. Evid Based Complement Alternat Med 2012; 2012:  Abd El Kader MA, El-Sammad NM, Fyiad AA. Effect of grape seeds extract in the modulation of matrix metalloproteinase-9 activity and oxidative stress induced by doxorubicin in mice. Life Sci J 2011; 8:  Ahmed AAE, Fatani AJ. Protective effect of grape seeds proanthocyanidins against naphthalene-induced hepatotoxicity in rats. Saudi Pharm J 2007; 15:  Raina K, Singh RP, Agarwal R, Agarwal C. Oral grape seed extract inhibits prostate tumor growth and progression in TRAMP mice. Cancer Res 2007; 67(12):  Bagchi D, Bagchi M, Stohs SJ, Das DK, Ray SD, Kuszynski CA, et al. Free radicals and grape seed proanthocyanidin extract: importance in human health and disease prevention. Toxicology 2000; 148(2-3):  Hedges SB. The origin and evolution of model organisms. Nat Rev Genet 2002; 3(11):  Singh RK, Bansode FW. Benzene-induced histopathological changes and germ cell population dynamics in testes of Sprague Dawley rats. J Environ Biol 2011; 32(6):  He LY, Xu SQ, Zhu HJ. Effects of benzene and selenium on telomerase in mouse lymphocytes in vivo. [Article in Chinese] Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi 2008; 26(7):  Fan XH. Effect of exposure to benzene on natural killer (NK) cell activity and interleukin-2 (IL-2) production of C57BL/6 mice. Nihon Ika Daigaku Zasshi 1992; 59(5):  Eastmond DA, Smith MT, Irons RD. An interaction of benzene metabolites reproduces the myelotoxicity observed with benzene exposure. Toxicol Appl Pharmacol 1987; 91(1):  Rozen MG, Snyder CA, Albert RE. Depressions in B- and T-lymphocyte mitogen-induced blastogenesis in mice exposed to low concentrations of benzene. Toxicol Lett 1984; 20(3):  Khan S, Priyamvada S, Khan SA, Khan W, Farooq N, et al. Effect of trichloroethylene (TCE) toxicity on the enzymes of carbohydrate metabolism, brush border membrane and oxidative stress in kidney and other rat tissues. Food Chem Toxicol 2009; 47(7):  Green JD, Snyder CA, LoBue J, Goldstein BD, Albert RE. Acute and chronic dose/response effect of benzene inhalation on the peripheral blood, bone marrow, and spleen cells of CD-1 male mice. Toxicol Appl Pharmacol 1981; 59(2):  Yalçin E, Oruç E, Cavuşoğlu K, Yapar K. Protective role of grape seed extract against doxorubicin-induced cardiotoxicity and genotoxicity in albino mice. J Med Food 2010; 13(4):  Graham JM. Homogenization of tissues and cells. In Subcellular Fractionation A Practical Approach. 1997; pp.1-29, Oxford University Press, Oxford, U.K.  Beutler E, Duran O, Kelley BM. Improved method for determination of blood glutathione. J Lab Clin Med 1963; 28:  Yoshioka T, Kawada K, Shimada T, Mori M. Lipid peroxidation in maternal and cord blood and protective mechanism against activated-oxygen toxicity in the blood. Am J Obstet Gynecol 1979; 135(3):  Yapar K, Cavuşoğlu K, Oruç E, Yalçin E. Protective role of Ginkgo biloba against hepatotoxicity and nephrotoxicity in uranium-treated mice. J Med Food 2010; 13(1):  Mansour HA, Newairy AS, Yousef MI, Sheweita SA. Biochemical study on the effects of some Egyptian herbs in alloxan-induced diabetic rats. Toxicology 2002; 170(3):  Raj HG, Malik S, Parmar VS, Kohli E, Tyagi YK, et al. Chemoprevention of benzene-induced bone marrow and pulmonary genotoxicity. Teratog Carcinog Mutagen 2001; 21(2):  Onunogbo CC, Ohaeri OC, Eleazu CO. Effect of mistletoe (viscum album) extract on the blood glucose, liver enzymes and electrolyte balance in alloxan induced diabetic rats. Am J Biochem Mol Biol 2013; 3:  Yusufi AN, Murayama N, Gapstur SM, Szczepanska-Konkel M, Dousa TP. Differential properties of brush-border membrane vesicles from early and late proximal tubules of rat kidney. Biochim Biophys Acta 1994; 1191(1):  Lall SB, Das N, Rama R, Peshin SS, Khattar S, et al. Cadmium induced nephrotoxicity in rats. Indian J Exp Biol 1997; 35(2): Turk J Biochem 2015; 40(1): Çavuşoğlu et al.
8  Grotto D, Maria LS, Valentini J, Paniz C, Garcia GSSC, et al. Importance of the lipid peroxidation biomarkers and methodological aspects for malondialdehyde quantification. Quim Nova 2009; 32:  Dündarz MR, Türkbay T, Akay C, Sarici SU, Aydin A, et al. Antioxidant enzymes and lipid peroxidation in adolescents with inhalant abuse. Turk J Pediatr 2003; 45(1):43-5.  Chunhua W, Di X, Hong Z, Ping G. Effect of lipid peroxidation in benzene in halation of mice. Acta Aca Med Bengbu 1997; 22:  Mohamed HA, El-Sayed IH, Moawad M. Protective effect of Nigella sativia seeds against dimethylaminoazobenzene (DAB) induced liver carcinogenesis. Nature Sci 2010; 8:80-7.  Ozden S, Alpertunga B. Effects of methiocarb on lipid peroxida- tion and glutathione level in rat tissues. Drug Chem Toxicol 2010; 33(1):50-4.  Jones DP. Redox potential of GSH/GSSG couple: assay and biological significance. Methods Enzymol 2002; 348:  Feng Y, Liu YM, Fratkins JD, LeBlanc MH. Grape seed extract suppresses lipid peroxidation and reduces hypoxic ischemic brain injury in neonatal rats. Brain Res Bull 2005; 66(2):  Balu M, Sangeetha P, Haripriya D, Panneerselvam C. Rejuvenation of antioxidant system in central nervous system of aged rats by grape seed extract. Neurosci Lett 2005; 383(3):  Sugisawa A, Inoue S, Umegaki K. Grape seed extract prevents H(2)O(2)-induced chromosomal damage in human lymphoblastoid cells. Biol Pharm Bull 2004; 27(9): Turk J Biochem 2015; 40(1): Çavuşoğlu et al.
International Journal of PharmTech Research CODEN( USA): IJPRIF ISSN : 0974-4304 Vol.1, No.3, pp 870-874, July-Sept 2009 CLINICAL STUDY OF MARKETED AYURVEDIC PREPARATION IN THYROID INDUCED STRESS A. A.
Guidance for Industry Safety Testing of Drug Metabolites U.S. Department of Health and Human Services Food and Drug Administration Center for Drug Evaluation and Research (CDER) February 2008 Pharmacology
The Physiology of Hyperbaric Oxygen Therapy Free Radicals and Reactive Oxygen Species I. Introduction Definition, Source, function and Purpose A. Definition of free radicals and reactive oxygen species
EUROPEAN COMMISSION HEALTH & CONSUMER PROTECTION DIRECTORATE-GENERAL Directorate C - Scientific Opinions C2 - Management of scientific committees II; scientific co-operation and networks Revision of the
PATIENT EDUCATION patienteducation.osumc.edu Blood delivers oxygen and nutrients to all the parts of the body. It also carries away waste products. The heart pumps blood in your body through a system of
Polyphenols in your diet may regulate food intake Role of dietary polyphenols in food intake Frontier Voice of Nutrition Remarks (May 06, 2013) Nalin Siriwardhana, Ph.D., interviewed Dr. Kiran Panickar,
Antioxidant Assay Kit Catalog Number KA1622 100 assays Version: 02 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use... 3 Principle of the Assay... 3 General
Endotoxicity and Cytotoxicity Studies 1) Aug 2005 In Vitro Chromosomal Aberration Study Cytotoxicity (File: TS39) Chinese Hamster Ovary (CHO) cells were grown in monolayer with and without metabolic activation.
Toxic Chemicals- Safe Work Practices Chemicals used in laboratories have a wide range of physical, chemical and toxicological properties that lead to adverse health effects in humans. The risks associated
Glutathione and Oxidative Stress - Part I By: James L. Holly, MD Oxidative Stress refers to effects from endogenous (produced in the body) toxins (free radicals) produced in the body by normal metabolism
Basic Overview of Preclinical Toxicology Animal Models Charles D. Hebert, Ph.D., D.A.B.T. December 5, 2013 Outline Background In Vitro Toxicology In Vivo Toxicology Animal Models What is Toxicology? Background
Available online at wwwpelagiaresearchlibrarycom European Journal of Experimental Biology, 2013, 3(2):280-284 ISSN: 2248 9215 CODEN (USA): EJEBAU Comparative Levels of ALT, AST, ALP and GGT in Liver associated
Smoking and Lung Cancer Objectives! Students should be able to identify some of the other consequences of smoking other than lung cancer! Students should be able to identify the effects of the chemicals
How to Interpret and Use Them René Romero, M.D. Clinical Director, Pediatric Hepatology CPG Gastroenterology, Hepatology and Nutrition Emory University School of Medicine Objectives Understand the anatomy
BISPHENL A DIGLYCIDYL ETHER Data were last evaluated in IARC (1989). 1. Exposure Data 1.1 Chemical and physical data 1.1.1 Nomenclature Chem. Abstr. Services Reg. No.: 1675-54-3 Systematic name: 2,2 -[(1-Methylethylidene)bis(4,1-phenyleneoxymethylene)]bis-
Islamic University-Gaza Faculty of Health Science The Medical laboratory science By: Hend N. Esawwaf & Ikram M. Ihmaid Supervisors: Dr. Abderaouf A. Elmanama Ms. Noor E. S. Abu Tayyem ه 1435 م May 2014
Hepatitis C Laboratory Tests and Hepatitis C If you have hepatitis C, your doctor will use laboratory tests to check your health. This handout will help you understand what the major tests are and what
EFFECT OF WATER-EXTRACTED PROPOLIS ON THE ACCUMULATION OF CHOLESTEROL INDUCED BY HIGH-CHOLESTEROL DIET IN THE RAT M.S. KWON*, Z.Z. HAN*, S.Y. PARK*, Y.S. CHOI*, H. LIM*, S.D. KIM 2, H.C. KIM 1 1 Neurotoxicology
Rodent Hepatic Carcinogens: Mode of Action: Framework (IPCS/ILSI) 1. Is the Weight of Evidence Sufficient to Establish a MoA in Animals? 2. Is the Animal MoA Plausible in Humans? 3. Taking Into Account
THE EFFECT OF SODIUM CHLORIDE ON THE GLUCOSE TOLERANCE OF THE DIABETIC RAT* BY JAMES M. ORTEN AND HENRY B. DEVLINt (From the Deparkment of Physiological Chemistry, Wayne University College of Medicine,
Section 4 Toxicology Occupational Health Any chemical you use incorrectly can result in over-exposure leading to adverse health effects immediately or in the future. All hazardous materials can be handled
Product Manual Lipid Extraction Kit (Chloroform Free), Trial Size Catalog Number STA-612-T 10 preps FOR RESEARCH USE ONLY Not for use in diagnostic procedures Introduction Lipids are a diverse group of
The Impact of Polyphenolic Compounds on Apoptosis in Epithelial Cells Mayson Husband, 12 th grade, Latta High School Abstract The purpose of this investigation was to determine whether polyphenolic compounds
WBC 4.00-10.50 WBC measures the total number of white blood cells in a given volume of blood. Since white blood cells kill bacteria, this count is a measure of the body s response to infection. In general,
Biological importance of metabolites Safety and efficacy aspects Bernard Walther Technologie Servier Biological importance of metabolites Safety testing of drug metabolites Bioanalytical strategy Structural
Chlorothalonil 89 5.7 CHLOROTHALONIL (081) TOXICOLOGY Chlorothalonil is the ISO approved common name for tetrachloroisophthalonitrile. Chlorothalonil (CAS No. 1897-45-6) is a non-systemic foliar fungicide
UNIVERSITY VETERINARY HOSPITAL & DIAGNOSTIC CENTER 952 East 900 South Salt Lake City, Utah 84105 (801) 596-9005 CHRONIC KIDNEY INSUFFICIENCY IN CATS By definition, kidney insufficiency or failure is the
Give Your Pet a Longer, Healthier Life Wellness Exam A wellness exam starts with a comprehensive physical examination by your pet s veterinarian. Once the exam has been completed, further services such
PR110 G-Biosciences 1-800-628-7730 1-314-991-6034 firstname.lastname@example.org A Geno Technology, Inc. (USA) brand name Dot Blot Analysis Teacher s Guidebook (Cat. # BE 502) think proteins! think G-Biosciences
This is a provisional English translation of an excerpt from the original full report. Risk Assessment Report Nitrate and Nitrite Nitrogen (beverages) Food Safety Commission of Japan (FSCJ) October 2012
Nursing 113 Pharmacology Principles 1. The study of how drugs enter the body, reach the site of action, and are removed from the body is called a. pharmacotherapeutics b. pharmacology c. pharmacodynamics
The Need for a PARP in vivo Pharmacodynamic Assay Jay George, Ph.D., Chief Scientific Officer, Trevigen, Inc., Gaithersburg, MD For further infomation, please contact: William Booth, Ph.D. Tel: +44 (0)1235
Steatosis Colorimetric Assay Kit Item No. 10012643 www.caymanchem.com Customer Service 800.364.9897 Technical Support 888.526.5351 1180 E. Ellsworth Rd Ann Arbor, MI USA TABLE OF CONTENTS GENERAL INFORMATION
Preventive Telemedicine Challenges in Russia Sergey Koldybaev Siberian Branch of Russian Academy of Sciences E mail: email@example.com Features of Russian Healthcare Most of qualified physicians concentrate
Glucose 65-99 mg/dl Glucose measures the amount of sugar in your blood. Glucose is the primary energy source in your body. Insulin (a natural hormone produced by the pancreas) controls how glucose is used
Human Clinical Study for Free Testosterone & Muscle Mass Boosting GE Nutrients, Inc. 920 E. Orangethorpe Avenue, Suite B Anaheim, California 92801, USA Phone: +1-714-870-8723 Fax: +1-732-875-0306 Contact
Laboratory Monitoring of Adult Hospital Patients Receiving Parenteral Nutrition Copy 1 Location of copies Web based only The following guideline is for use by medical staff caring for the patient and members
JOINT FAO/WHO MEETING ON PESTICIDE RESIDUES Geneva, 9 13 May 2016 SUMMARY REPORT Issued 16 May 2016 Edited versions of these evaluations and general considerations will be published in the report of the
Nonalcoholic Steatohepatitis National Digestive Diseases Information Clearinghouse National Institute of Diabetes and Digestive and Kidney Diseases NATIONAL INSTITUTES OF HEALTH Nonalcoholic steatohepatitis
TOXIC CHEMICALS Supawan Tantayanon Department of Chemistry, Faculty of Science Chulalongkorn University Toxic Chemicals Any chemical when ingested, inhaled, or absorbed, or when applied to, injected into
FOR YOUR PATIENTS WITH RELAPSING FORMS OF MS INITIATING ORAL AUBAGIO (teriflunomide) THERAPY WARNING: HEPATOTOXICITY AND RISK OF TERATOGENICITY Severe liver injury including fatal liver failure has been
Your Lab Report Understanding the Results When your doctor orders lab tests as a part of your care, you may receive a written copy of the report or you may be able to view your results in a patient portal
2011 Health Risk Limits for Groundwater Health Risk Assessment Unit, Environmental Health Division 651-201-4899 651-201-5797 TDD Web Publication Date: March 21, 2011 Chemical Name: Acetone CAS: 67-64-1
LIVER FUNCTION TESTS University of PNG School of Medicine and Health Sciences, Division of Basic Medical Sciences, Discipline of Biochemistry & Molecular Biology VJ Temple 1 What are some of the functions
Vitamin D und seine Bedeutung im Immunsystem und bei der Infektabwehr Stefan Pilz Department of Internal Medicine, Division of Endocrinology and Metabolism, Medical University of Graz, Austria Department
Effect of Fish Oil on High Lipid Fed Albino Rats *Luka, C.D and **Mohammed, A. *Department of Biochemistry, Faculty of Medical Science, University of Jos **Department of Biochemistry, Faculty of Pure &
Antioxidant Products A comprehensive range of ALB BIL FERR GR Ransel Ransod antioxidant TAS testing TIBC kits TF BIL FERR GR Ransel Ransod TAS TIBC TF UA A FERR GR Ransel Ransod TAS TIBC TF UA ALB GR Ransel
Are 50/60 Hz magnetic fields carcinogenic to humans? Michel Plante M.D. Magnetic fields are present everywhere in our environment. They are commonly measured in Tesla (T), the international unit for magnetic
RICEO TM Water Soluble Concentrate Powder Providing Natural Vitamin B Group and Minerals RICEO TM is a water-soluble powder extracted from defatted rice bran. It is mainly composed of carbohydrate, dietary
Module Three Risk Assessment 136 Module Three Introduction to Risk Assessment Time Allotted: 90 Minutes Objectives: Upon completion of this module, the learner will be able to # Define and understand the
174 CHAPTER 9 ANTI-CANCER ACTIVITY OF COMPOUND-III 9.1 IINTRODUCTION Oleanolic acid is a naturally occurring triterpenoid, widely distributed in food and medicinal plants. It is relatively non-toxic and
The basic layout, the main functions and instrumentation concept of micro Inspection Division laboratory, 1, Virology Laboratory 1. Functions: for the city to monitor the prevalence of HIV disease, dealing
LOOK, FEEL AND LIVE BETTER Topical Skin Care Pycnogenol in Topical Skin Care Pycnogenol is widely used in topical and oral applications for various dermatological indications. A unique combination of pharmacological
Defining pediatric reference ranges in laboratory medicine: Why adult ranges don t always work Children are not small adults Michael J. Bennett PhD, FRCPath, FACB DABCC University of Pennsylvania & Children
D. Vitamin D. Two main forms; vitamin D2 and D3 H H D3 - Cholecalciferol D2 - Ergocalciferol Technically, vitamin D is not a vitamin. It is the name given to a group of fat-soluble prohormones (substances
Pharmacokinetic Test of Calcium Aspartate Anhydrous This study tests the metabolic mechanism and the pharmacokinetics on the changes of calcium aspartate anhydrous in animal bodies over time. The results
NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES Division of Extramural Research and Training NATIONAL ADVISORY ENVIRONMENTAL HEALTH SCIENCES COUNCIL May 12-13, 2010 Concept Clearance Small Business
Effect of on Blood Glucose Levels in Alloxan-induced Diabetic Mice Kathryn Niedzielski Advisor: Dr. Linda Swift ABSTRACT Diabetes is a condition in the body where the pancreas does not produce enough insulin
1. Identification of Substance/Preparation and of the Company Trade name : Company : Paksoy Kimyevi Maddeler San. ve Tic. A.Ş. GOSB 700. sokak No:720 41480 Gebze/ TURKEY Phone number : 00 90 262 751 13
Tropical Agricultural Research Vol. 26 (4): 584 595 (20) Changes in the Serum Enzyme Levels and Liver Lesions of Broiler Birds Reared Under Different Management Conditions S.S.H.M.M.L. Senanayake, J.G.S.
RELATIONSHIP BETWEEN BIOCHEMICAL PARAMETERS - LIVER ENZYMES (AST, ALT AND ALP) AND BLOOD PHENYTOIN LEVELS IN PATIENTS OF EPILEPSY Ashok Binjawadgi 1, Patil B V 2, Shrinivas Raikar 3, Manu Bhardwaj 4, Basavambika
Coding and Payment Guide for Laboratory Services An essential coding, billing, and payment resource for laboratory and pathology services Contents Introduction............................... 1 Coding Systems...............................
Free radicals - Enemies of health, beauty and youth Free radicals (FR) are natural molecules produced in the body following biochemical reactions related to the behaviour of singlet oxygen. The human body
LGD 6972: Glucagon Receptor Antagonist Highly Potent, Orally Bioavailable, Small Molecule Glucagon Receptor Antagonist for the Treatment of Type 2 Diabetes Overview Diabetes is one of the largest and fastest
Support Program for Improving Graduate School Education Advanced Education Program for Integrated Clinical, Basic and Social Medicine January 27, 2009 Dear Professors (representative) of departments, Subject:
Antioxidants and Herbs Presented at the ACHS 3rd Annual Herb Day Celebration 2008 What are antioxidants? Why are they useful in our bodies and herbal medicine? Where are they found in herbs? Antioxidants?
Greyhound Sports Nutrition Introduction Nutrition has the potential to have a real impact on the performance of racing greyhounds. There is evidence in both dogs and human athletes of the importance of
Regulating the Internal Environment Water Balance & Nitrogenous Waste Removal 2006-2007 Animal systems evolved to support multicellular life CH CHO O 2 O 2 NH 3 CH CHO O 2 CO 2 NH NH 3 O 2 3 NH 3 intracellular
Chapter 3 Vitamin C in Plasma and Leukocytes of Camels with Different Reproductive Status H.E. Mohamed 1, H.M. Mousa 1 and A.C. Beynen 2 1 Department of Biochemistry, Faculty of Veterinary Science, University
Effect of Crude Aloe vera Gel on Serum Enzymes, Proteins and Liver Histology in Alloxan - Induced Diabetic Rats Akpan, U. P. 1, Nna, V. U. 2, Ikpi, D. E. 3, Osim E. E. 4, Antai, A. B. 5 1 Department of
What Do Those Lab Tests Mean? This information is not meant to be a substitute for veterinary care. Always follow the instructions provided by your veterinarian. The results of laboratory tests on a patient
77:222 Spring 2003 Free Radicals in Biology and Medicine Page 0 This student paper was written as an assignment in the graduate course Free Radicals in Biology and Medicine (77:222, Spring 2003) offered
Laboratory 12 Homeostasis (LM pages 157 171) Time Estimate for Entire Lab: 1.5 hours Seventh Edition Changes This was lab 11 in the previous edition. Section 12.3 Kidneys was reorganized; Nephron Structure
Shantaram S. Joshi, Ph.D. Functional Genomics of Leukemia Dendritic Cell Based Therapy for Cancer Dept. of Genetics, Cell Biology and Anatomy University of Nebraska Medical Center Omaha, Nebraska 68198-6395
Chapter 12: SPECIFIC TARGET ORGAN SYSTEMIC TOXICITY (TOST) FOLLOWING A SINGLE EXPOSURE DEFINITIONS 1. Classification identifies the chemical substance as being a specific target organ/systemic toxicant
Molecular mechanisms of carcinogenesis Nik Hodges School of Biosciences N.Hodges@bham.ac.uk Aims What is cancer at the cellular level? How do chemical and physical agents cause cancer? How do we test for
Guidance for Industry S9 Nonclinical Evaluation for Anticancer Pharmaceuticals U.S. Department of Health and Human Services Food and Drug Administration Center for Drug Evaluation and Research (CDER) Center
MATERIAL SAFETY DATA SHEET DATE: 2009, revised July 2011 PRODUCT NAME: BOSTON BRAKE & PARTS CLEANER PRODUCT CODE: 78200 SUPPLIER S NAME : SECTION I SUPPLIER IDENTIFICATION CAMPBELLS WHOLESALE PTY LTD ADDRESS
Orange County 1. 1. Where Where does does the the Medical Laboratory Technician Work Work? 2. 2. Why Why you you should should become a Med. Med. Lab. Lab. Tech. Tech. 3. 3. Overview of of Core Core Courses
Course Curriculum for Master Degree in Food Hygiene /Faculty of Veterinary Medicine The Master Degree Food Hygiene /Veterinary Medicine is awarded by the Faculty of Graduate Studies at Jordan University
Adams Memorial Hospital Decatur, Indiana EXPLANATION OF LABORATORY TESTS Your health is important to us! The test descriptions listed below are for educational purposes only. Laboratory test interpretation