Conclusion indicates that gold(III) complexes have good potential for the treatment of cancer. In addition [Au(en)Cl2]+ complex shows cytotoxicity profiles comparable to cisplatin [27]. This study has led us to investigate further the conclusion achieved by the in vitro studies of Milovanovic et al [27]. The title compound 
is a newly developed gold (III) compound [Au(en)Cl2]Cl, gold complexed with N-substituted ethylenediamine. (Fig.1). It has been prepared and fully characterized by spectroscopic techniques such as UV is, Far-IR, IR spectroscopy, solution, Xray and solid NMR. The solution NMR was measured in D2O, implicating that it is water soluble [28,29]. In the current study we evaluated the histopathological toxicity of this compound in renal and hepatic order FCCP tissues of rats.Acute Toxicity StudyIn acute toxicity, 5 groups of rats (A/I-E/I), with each group comprising 5 animals, were administered gold compound intraperitoneally in doses of 1500 mg/kg, 750 mg/kg, 375 mg/kg, 187.5 mg/kg and 93.75 mg/kg, respectively. A control group of 5 animals (F/I) was simultaneously administered 0.2 ml water intraperitoneally. After 24 hours, the number of deceased rats was counted in each group and LD50 (dose that kills 50 of animals) was calculated (322 mg/kg) by the method of Miller and Tainter [30]. Autopsy was carried out in all animals and renal as well as hepatic tissues were preserved in 10 buffered formalin for subsequent evaluation of histopathological alterations.Sub-acute Toxicity StudyThe rats in this component of the study were divided into two treatment groups, A/II and B/II, with six rats in each. Group “A/ II” served as the experimental group while group “B/II” served as the control. Rats in the experimental group (A/II) were injected with 32.2 mg/kg (1/10 of LD50) body weight of the gold compound while rats in the control group (B/II) were injected with normal saline daily for 14 days. Autopsy was carried out in all the rats. 
Renal and hepatic tissues were preserved in 10 buffered formalin until subjected to histopathological evaluation.Histopathological Work Upa) Fixation and tissue processing. The formalin preserved hepatic and renal tissue samples of [Au(en)Cl2]Cl dosed rats and controls were processed in an automated tissue processor (Tissue?tek VIP-5, from SAKURA). The processing consisted of an initial 2 step fixation comprising tissue immersion in 10 buffered formalin for two hours each, followed by removal of fixative in distilled water for 30 minutes. Dehydration was then 1326631 carried out by running the tissues through a graded series of alcohol (70 , 90 , and 100 ). The tissue was initially exposed to 70 alcoholTable 1. Histological categorization of drug-induced hepatic lesions.Acute hepatitis and cholestatic hepatitis Acute liver failure Cholestasis Chronic Hepatitis Granulomatous hepatitis Steatosis/Steatohepatitis Vascular Abnormalities doi:10.1371/journal.pone.0051889.t001 ��-Sitosterol ��-D-glucoside biological activity Macrovesicular, Microvesicular, Steatohepatitis Sinusoidal obstruction syndrome Necrosis with marked inflammation, Necrosis with little or no inflammation Bland cholestasis, Cholestatic hepatitisRenal and Hepatic Toxicity of a Gold (III) CompoundSinusoidal Obstruction syndromefor 30 minutes followed by 90 alcohol for 1 hour and then two cycles of absolute alcohol, each for one hour. Dehydration was then followed by clearing the samples in several changes of xylene. It consisted of tissue immersion for an hour in a mixture comprising 50 alcohol.Conclusion indicates that gold(III) complexes have good potential for the treatment of cancer. In addition [Au(en)Cl2]+ complex shows cytotoxicity profiles comparable to cisplatin [27]. This study has led us to investigate further the conclusion achieved by the in vitro studies of Milovanovic et al [27]. The title compound is a newly developed gold (III) compound [Au(en)Cl2]Cl, gold complexed with N-substituted ethylenediamine. (Fig.1). It has been prepared and fully characterized by spectroscopic techniques such as UV is, Far-IR, IR spectroscopy, solution, Xray and solid NMR. The solution NMR was measured in D2O, implicating that it is water soluble [28,29]. In the current study we evaluated the histopathological toxicity of this compound in renal and hepatic tissues of rats.Acute Toxicity StudyIn acute toxicity, 5 groups of rats (A/I-E/I), with each group comprising 5 animals, were administered gold compound intraperitoneally in doses of 1500 mg/kg, 750 mg/kg, 375 mg/kg, 187.5 mg/kg and 93.75 mg/kg, respectively. A control group of 5 animals (F/I) was simultaneously administered 0.2 ml water intraperitoneally. After 24 hours, the number of deceased rats was counted in each group and LD50 (dose that kills 50 of animals) was calculated (322 mg/kg) by the method of Miller and Tainter [30]. Autopsy was carried out in all animals and renal as well as hepatic tissues were preserved in 10 buffered formalin for subsequent evaluation of histopathological alterations.Sub-acute Toxicity StudyThe rats in this component of the study were divided into two treatment groups, A/II and B/II, with six rats in each. Group “A/ II” served as the experimental group while group “B/II” served as the control. Rats in the experimental group (A/II) were injected with 32.2 mg/kg (1/10 of LD50) body weight of the gold compound while rats in the control group (B/II) were injected with normal saline daily for 14 days. Autopsy was carried out in all the rats. Renal and hepatic tissues were preserved in 10 buffered formalin until subjected to histopathological evaluation.Histopathological Work Upa) Fixation and tissue processing. The formalin preserved hepatic and renal tissue samples of [Au(en)Cl2]Cl dosed rats and controls were processed in an automated tissue processor (Tissue?tek VIP-5, from SAKURA). The processing consisted of an initial 2 step fixation comprising tissue immersion in 10 buffered formalin for two hours each, followed by removal of fixative in distilled water for 30 minutes. Dehydration was then 1326631 carried out by running the tissues through a graded series of alcohol (70 , 90 , and 100 ). The tissue was initially exposed to 70 alcoholTable 1. Histological categorization of drug-induced hepatic lesions.Acute hepatitis and cholestatic hepatitis Acute liver failure Cholestasis Chronic Hepatitis Granulomatous hepatitis Steatosis/Steatohepatitis Vascular Abnormalities doi:10.1371/journal.pone.0051889.t001 Macrovesicular, Microvesicular, Steatohepatitis Sinusoidal obstruction syndrome Necrosis with marked inflammation, Necrosis with little or no inflammation Bland cholestasis, Cholestatic hepatitisRenal and Hepatic Toxicity of a Gold (III) CompoundSinusoidal Obstruction syndromefor 30 minutes followed by 90 alcohol for 1 hour and then two cycles of absolute alcohol, each for one hour. Dehydration was then followed by clearing the samples in several changes of xylene. It consisted of tissue immersion for an hour in a mixture comprising 50 alcohol.
