BioMed Research International: Toxicology The latest articles from Hindawi © 2017 , Hindawi Limited . All rights reserved. Mutagenic and Cytotoxicity LQB 123 Profile: Safety and Tripanocidal Effect of a Phenyl-t-Butylnitrone Derivative Wed, 15 Feb 2017 07:12:05 +0000 The therapeutic options for Chagas disease are limited and its treatment presents a number of drawbacks including toxicity, drug resistance, and insufficient effectiveness against the chronic stage of the disease. Therefore, new therapeutical options are mandatory. In the present work, we evaluated the effect of a phenyl-tert-butylnitrone (PBN) derivate, LQB 123, against Trypanosoma cruzi forms. LQB 123 presented a trypanocidal effect against bloodstream trypomastigotes (IC50 =  μM) and intracellular amastigotes infecting peritoneal macrophages (IC50 =  μM), with no harmful effects upon the mammalian cells (CC50 values greater than 4 mM), resulting in a high selectivity index (CC50/IC50 > 20). Additionally, metacyclic trypomastigotes submitted to LQB 123 presented an IC50 of about  μM and epimastigotes forms incubated with different concentrations of LQB 123 presented an inhibition of parasite growth with an IC50 of  μM. Finally, we investigated the mutagenic potential of the nitrone by the Salmonella/microsome assay and observed no induction of mutagenicity even in concentrations as high as 33000 μM. Taken together, these results present a nonmutagenic compound, with trypanocidal activity against all relevant forms of T. cruzi, offering new insights into CD treatment suggesting additional in vivo tests. Mauricio Peixoto Cupello, Francis Monique Saraiva, Pedro Ippolito, Andréia da Silva Fernandes, Rubem Figueiredo Sadoko Menna-Barreto, Debora de Sousa dos Santos Costa, Jessica Isis Oliveira Paula, Paulo Roberto Ribeiro Costa, Natália Peleira Nogueira, Israel Felzenswalb, Ayres Guimarães Dias, and Marcia Cristina Paes Copyright © 2017 Mauricio Peixoto Cupello et al. All rights reserved. Bisphenol A Is More Potent than Phthalate Metabolites in Reducing Pancreatic β-Cell Function Mon, 13 Feb 2017 00:00:00 +0000 Bisphenol A (BPA) and phthalates are common environmental contaminants that have been proposed to influence incidence and development of types 1 and 2 diabetes. Thus, effects of BPA and three phthalate metabolites (monoisobutyl phthalate (MiBP), mono-n-butyl phthalate (MnBP), and mono-(2-ethylhexyl) phthalate (MEHP)) were studied in the pancreatic β-cell line INS-1E, after 2–72 h of exposure to 5–500 μM. Three endpoints relevant to accelerated development of types 1 or 2 diabetes were investigated: β-cell viability, glucose-induced insulin secretion, and β-cell susceptibility to cytokine-induced cell death. BPA and the phthalate metabolites reduced cellular viability after 72 h of exposure, with BPA as the most potent chemical. Moreover, BPA, MEHP, and MnBP increased insulin secretion after 2 h of simultaneous exposure to chemicals and glucose, with potency BPA > MEHP > MnBP. Longer chemical exposures (24–72 h) showed no consistent effects on glucose-induced insulin secretion, and none of the environmental chemicals affected susceptibility to cytokine-induced cell death. Overall, BPA was more potent than the investigated phthalate metabolites in affecting insulin secretion and viability in the INS-1E pancreatic β-cells. In contrast to recent literature, concentrations with relevance to human exposures (1–500 nM) did not affect the investigated endpoints, suggesting that this experimental model displayed relatively low sensitivity to environmental chemical exposure. Nina Mickelson Weldingh, Lena Jørgensen-Kaur, Rune Becher, Jørn A. Holme, Johanna Bodin, Unni C. Nygaard, and Anette Kocbach Bølling Copyright © 2017 Nina Mickelson Weldingh et al. All rights reserved. Predicting Presynaptic and Postsynaptic Neurotoxins by Developing Feature Selection Technique Sun, 12 Feb 2017 09:24:54 +0000 Presynaptic and postsynaptic neurotoxins are proteins which act at the presynaptic and postsynaptic membrane. Correctly predicting presynaptic and postsynaptic neurotoxins will provide important clues for drug-target discovery and drug design. In this study, we developed a theoretical method to discriminate presynaptic neurotoxins from postsynaptic neurotoxins. A strict and objective benchmark dataset was constructed to train and test our proposed model. The dipeptide composition was used to formulate neurotoxin samples. The analysis of variance (ANOVA) was proposed to find out the optimal feature set which can produce the maximum accuracy. In the jackknife cross-validation test, the overall accuracy of 94.9% was achieved. We believe that the proposed model will provide important information to study neurotoxins. Hua Tang, Yunchun Yang, Chunmei Zhang, Rong Chen, Po Huang, Chenggang Duan, and Ping Zou Copyright © 2017 Hua Tang et al. All rights reserved. Pharmacokinetic and Toxicological Evaluation of a Zinc Gluconate-Based Chemical Sterilant Using In Vitro and In Silico Approaches Thu, 19 Jan 2017 12:49:27 +0000 Sclerosing agents as zinc gluconate-based chemical sterilants (Infertile®) are used for chemical castration. This solution is injected into the animal testis, but there are not enough evidences of its safety profiles for the receivers. The present work aimed to establish the pharmacokinetics and toxicological activity of Infertile, using in vitro and in silico approaches. The evaluation at the endpoint showed effects in a dose-dependent manner. Since necrosis is potentially carcinogenic, the possible cell death mechanism could be apoptosis. Our data suggested that Infertile at 60 mM presented risk for animal health. Even though Infertile is a licensed product by the Brazilian Ministry of Agriculture, Livestock and Supply, it presented a high mutagenic potential. We suggest that the optimal dose must be less than 6 mM, once, at this concentration, no mutagenicity or genotoxicity was observed. Carlos F. Araujo-Lima, Rafael J. M. Nunes, Raphael M. Carpes, Claudia A. F. Aiub, and Israel Felzenszwalb Copyright © 2017 Carlos F. Araujo-Lima et al. All rights reserved. Toxicity Studies of Ethyl Maltol and Iron Complexes in Mice Thu, 19 Jan 2017 10:20:00 +0000 Ethyl maltol and iron complexes are products of ethyl maltol and the iron found in the cooking pots used to prepare the Chinese dish, hot-pot. Because their safety is undocumented, the toxicity study of ethyl maltol and iron complexes was conducted in male and female Kunming (KM) mice. The animal study was designed based on the preliminary study conducted to determine the median lethal dose (LD50). The doses used in the study were 0, 1/81, 1/27, 1/9, and 1/3 of the LD50 (mg kg body weight (BW)−1 day−1) dissolved in the water. The oral LD50 of the ethyl maltol and iron complexes was determined to be 743.88 mg kg BW−1 in mice. The ethyl maltol and iron complexes targeted the endocrine organs including the liver and kidneys following the 90 D oral exposure. Based on the haematological data, the lowest-observed-adverse-effect level (LOAEL) of the ethyl maltol and iron complexes was determined to be 1/81 LD50 (9.18 mg kg BW−1 day−1) in both male and female mice. Therefore, we suggest that alternative strategies for preparing the hot-pot, including the use of non-Fe-based cookware, need to be developed and encouraged to avoid the formation of the potentially toxic complexes. Zhen Li, Jieli Lu, Chonghui Wu, Quanhai Pang, Zhiwei Zhu, Ruipeng Nan, Ruochen Du, and Jia Chen Copyright © 2017 Zhen Li et al. All rights reserved. Drug-Induced Liver Injury Mon, 09 Jan 2017 09:27:46 +0000 Eileen E. N. Almario, Jürgen Borlak, Ayako Suzuki, and Minjun Chen Copyright © 2017 Eileen E. N. Almario et al. All rights reserved. Idiosyncratic Drug-Induced Liver Injury (IDILI): Potential Mechanisms and Predictive Assays Wed, 04 Jan 2017 06:56:12 +0000 Idiosyncratic drug-induced liver injury (IDILI) is a significant source of drug recall and acute liver failure (ALF) in the United States. While current drug development processes emphasize general toxicity and drug metabolizing enzyme- (DME-) mediated toxicity, it has been challenging to develop comprehensive models for assessing complete idiosyncratic potential. In this review, we describe the enzymes and proteins that contain polymorphisms believed to contribute to IDILI, including ones that affect phase I and phase II metabolism, antioxidant enzymes, drug transporters, inflammation, and human leukocyte antigen (HLA). We then describe the various assays that have been developed to detect individual reactions focusing on each of the mechanisms described in the background. Finally, we examine current trends in developing comprehensive models for examining these mechanisms. There is an urgent need to develop a panel of multiparametric assays for diagnosing individual toxicity potential. Alexander D. Roth and Moo-Yeal Lee Copyright © 2017 Alexander D. Roth and Moo-Yeal Lee. All rights reserved. The Monitoring and Assessment of Aquatic Toxicology Tue, 03 Jan 2017 00:00:00 +0000 Zongming Ren, Tae-Soo Chon, Chunlei Xia, and Fengqing Li Copyright © 2017 Zongming Ren et al. All rights reserved. Aquatic Environmental Health and Toxicology Wed, 28 Dec 2016 06:50:15 +0000 Zhi-Hua Li, Kaiyu He, Chunsheng Liu, Ping Li, and Vladimir Zlabek Copyright © 2016 Zhi-Hua Li et al. All rights reserved. Regulation of Human Cytochrome P4501A1 (hCYP1A1): A Plausible Target for Chemoprevention? Mon, 26 Dec 2016 07:16:00 +0000 Human cytochrome P450 1A1 (hCYP1A1) has been an object of study due to its role in precarcinogen metabolism; for this reason it is relevant to know more in depth the mechanisms that rule out its expression and activity, which make this enzyme a target for the development of novel chemiopreventive agents. The aim of this work is to review the origin, regulation, and structural and functional characteristics of CYP1A1 letting us understand its role in the bioactivation of precarcinogen and the consequences of its modulation in other physiological processes, as well as guide us in the study of this important protein. Rebeca Santes-Palacios, Diego Ornelas-Ayala, Noel Cabañas, Ana Marroquín-Pérez, Alexis Hernández-Magaña, Sitlali del Rosario Olguín-Reyes, Rafael Camacho-Carranza, and Jesús Javier Espinosa-Aguirre Copyright © 2016 Rebeca Santes-Palacios et al. All rights reserved. Dietary Biotin Supplementation Modifies Hepatic Morphology without Changes in Liver Toxicity Markers Sun, 25 Dec 2016 14:06:25 +0000 Pharmacological concentrations of biotin have pleiotropic effects. Several reports have documented that biotin supplementation decreases hyperglycemia. We have shown that a biotin-supplemented diet increased insulin secretion and the mRNA abundance of proteins regulating insulin transcription and secretion. We also found enlarged pancreatic islets and modified islet morphology. Other studies have shown that pharmacological concentrations of biotin modify tissue structure. Although biotin administration is considered safe, little attention has been given to its effect on tissue structure. In this study, we investigated the effect of biotin supplementation on hepatic morphology and liver toxicity markers. Male BALB/cAnN Hsd mice were fed a control or a biotin-supplemented diet for 8 weeks. Versus the control mice, biotin-supplemented mice had an altered portal triad with dilated sinusoids, increased vascularity, and bile conducts. Furthermore, we observed an increased proportion of nucleomegaly and binucleated hepatocytes. In spite of the liver morphological changes, no differences were observed in the serum liver damage indicators, oxidative stress markers, or antioxidant enzymes. Our data demonstrate for the first time that biotin supplementation affects liver morphology in normal mice, and that these modifications are not paralleled with damage markers. Leticia Riverón-Negrete, Gloria Sicilia-Argumedo, Carolina Álvarez-Delgado, Elvia Coballase-Urrutia, Jonathan Alcántar-Fernández, and Cristina Fernandez-Mejia Copyright © 2016 Leticia Riverón-Negrete et al. All rights reserved. Biodegradation of Dimethyl Phthalate by Freshwater Unicellular Cyanobacteria Mon, 19 Dec 2016 11:24:50 +0000 The biodegradation characteristics of dimethyl phthalate (DMP) by three freshwater unicellular organisms were investigated in this study. The findings revealed that all the organisms were capable of metabolizing DMP; among them, Cyanothece sp. PCC7822 achieved the highest degradation efficiency. Lower concentration of DMP supported the growth of the Cyanobacteria; however, with the increase of DMP concentration growth of Cyanobacteria was inhibited remarkably. Phthalic acid (PA) was detected to be an intermediate degradation product of DMP and accumulated in the culture solution. The optimal initial pH value for the degradation was detected to be 9.0, which mitigated the decrease of pH resulting from the production of PA. The optimum temperature for DMP degradation of the three species of organisms is 30°C. After 72 hours’ incubation, no more than 11.8% of the residual of DMP aggregated in Cyanobacteria cells while majority of DMP remained in the medium. Moreover, esterase was induced by DMP and the activity kept increasing during the degradation process. This suggested that esterase could assist in the degradation of DMP. Xiaohui Zhang, Lincong Liu, Siping Zhang, Yan Pan, Jing Li, Hongwei Pan, Shiguo Xu, and Feng Luo Copyright © 2016 Xiaohui Zhang et al. All rights reserved. Research Advances on Hepatotoxicity of Herbal Medicines in China Sun, 18 Dec 2016 09:24:39 +0000 In general, herbal medicines have been considered as safe by the general public, since they are naturally occurring and have been applied in treatment for over thousands of years. As the use of herbal medicine is rapidly increasing globally, the potential toxicity of herbal drugs, in particular drug-induced liver injury (DILI), has now become a serious medical issue. According to the literature, the authors analyzed and discussed the hepatotoxicity problem of Chinese herbal medicines (CHM), including global overview on herbal-induced liver injury (HILI), current research progress on toxic CHM, diagnosis and treatment of HILI, and modern approaches and technologies of study of hepatotoxicity. As to promote the recognition of HILI and tackle the issue, a guideline for the diagnosis and treatment of HILI has recently been drafted by Chinese scientists. As suggested by the guideline, the hepatotoxicity issue of CHM, as a matter of fact, is overestimated. Up to date, the investigation of hepatotoxicity of CHM is now booming with worldwide application of CHM. This review therefore provides useful information for investigating hepatotoxicity of herbal medicine and characterizing DILI caused by CHM. In addition, authors describe in which way further efforts should be made to study the rationale of CHM and liver injury. Changxiao Liu, Huirong Fan, Yazhuo Li, and Xiaohe Xiao Copyright © 2016 Changxiao Liu et al. All rights reserved. TAMH: A Useful In Vitro Model for Assessing Hepatotoxic Mechanisms Thu, 15 Dec 2016 13:23:00 +0000 In vitro models for hepatotoxicity can be useful tools to predict in vivo responses. In this review, we discuss the use of the transforming growth factor-α transgenic mouse hepatocyte (TAMH) cell line, which is an attractive model to study drug-induced liver injury due to its ability to retain a stable phenotype and express drug-metabolizing enzymes. Hepatotoxicity involves damage to the liver and is often associated with chemical exposure. Since the liver is a major site for drug metabolism, drug-induced liver injury is a serious health concern for certain agents. At the molecular level, various mechanisms may protect or harm the liver during drug-induced hepatocellular injury including signaling pathways and endogenous factors (e.g., Bcl-2, GSH, Nrf2, or MAPK). The interplay between these and other pathways in the hepatocyte can change upon drug or drug metabolite exposure leading to intracellular stress and eventually cell death and liver injury. This review focuses on mechanistic studies investigating drug-induced toxicity in the TAMH line and how alterations to hepatotoxic mechanisms in this model relate to the in vivo situation. The agents discussed herein include acetaminophen (APAP), tetrafluoroethylcysteine (TFEC), flutamide, PD0325901, lapatinib, and flupirtine. Madison Davis and Brendan D. Stamper Copyright © 2016 Madison Davis and Brendan D. Stamper. All rights reserved. Enhanced Protective Effects of Combined Treatment with β-Carotene and Curcumin against Hyperthermic Spermatogenic Disorders in Mice Mon, 05 Dec 2016 07:07:28 +0000 Scrotal hyperthermia leads to oxidative stress and apoptosis in spermatogenic cells, which subsequently causes male infertility. In this study, we examined the effects of β-carotene and/or curcumin on heat-stress- (HS-) induced testicular injuries in mice. ICR male mice (8 weeks old) were consecutively treated with β-carotene (10 mg/kg) and/or curcumin (20 mg/kg) orally once a day for 14 days and then subjected to single exposure with scrotal HS at 43°C for 15 min on day 7. HS induced a significant reduction in testicular weight, appearance of multinucleated giant cells, and desquamation of germ cells in destructive seminiferous tubules, as well as degenerative Leydig cells. Moreover, HS reduced the superoxide dismutase (SOD) activity and mRNA levels of mitochondrial SOD, phospholipid hydroperoxide glutathione peroxidase, B-cell lymphoma-extra-large, and 3β-hydroxysteroid dehydrogenase, with increases in lipid peroxidation levels and mRNA levels of BCL2-associated X protein and caspase-3 relative to those of the control group. However, these changes were significantly recovered by combined treatment with β-carotene and curcumin after HS. These findings indicate that the combined treatment with β-carotene and curcumin might be a valuable protective agent to ameliorate hyperthermic spermatogenic disorders via its potent antioxidative, antiapoptotic, and androgen synthetic effects. Chunmei Lin, Yun Seok Choi, Seul Gi Park, Lee Wha Gwon, Jong Geol Lee, Jung-Min Yon, In-Jeoung Baek, Beom Jun Lee, Young Won Yun, and Sang-Yoon Nam Copyright © 2016 Chunmei Lin et al. All rights reserved. Quantitative Characterization of the Toxicities of Cd-Ni and Cd-Cr Binary Mixtures Using Combination Index Method Thu, 01 Dec 2016 14:07:53 +0000 Direct equipartition ray design was used to construct Cd-Ni and Cd-Cr binary mixtures. Microplate toxicity analysis was used to evaluate the toxicity of individual substance and the Cd-Ni and Cd-Cr mixtures on Chlorella pyrenoidosa and Selenastrum capricornutum. The interacting toxicity of the mixture was analyzed with concentration addition (CA) model. In addition, combination index method (CI) was proposed and used to quantitatively characterize the toxicity of the binary mixtures of Cd-Ni and Cd-Cr observed in experiment and find the degree of deviation from the predicted outcome of the CA model, that is, the intensity of interacting toxicity. Results indicate that most of the 20 binary mixtures exhibit enhancing and synergistic effect, and only Cd-Cr-R4 and Cd-Cr-R5 mixtures have relatively high antagonistic effects against C. pyrenoidosa. Based on confidence interval, CI can compare the intensities of interaction of the mixtures under varying levels of effect. The characterization methods are applicable for analyzing binary mixture with complex interaction. Lingyun Mo, Minyi Zheng, Meng Qin, Xin Zhang, Jie Liu, Litang Qin, Honghu Zeng, and Yanpeng Liang Copyright © 2016 Lingyun Mo et al. All rights reserved. Micronucleated Erythrocytes in Peripheral Blood from Neonate Rats Exposed by Breastfeeding to Cyclophosphamide, Colchicine, or Cytosine-Arabinoside Mon, 28 Nov 2016 11:27:42 +0000 Genotoxic exposure to chemical substances is common, and nursing mothers could transmit harmful substances or their metabolites to their offspring through breast milk. We explored the possibility of determining genotoxic effects in the erythrocytes of breastfeeding rat pups whose mothers received a genotoxic compound while nursing. Ten groups of female rats and five pups per dam were studied. The control group received sterile water, and the experimental groups received one of three different doses of cyclophosphamide, colchicine, or cytosine-arabinoside. Blood smears were prepared from samples taken from each dam and pup every 24 h for six days. There were increased numbers of micronucleated erythrocytes (MNEs) and micronucleated polychromatic erythrocytes (MNPCEs) in the samples from pups in the experimental groups () and increased MNPCE frequencies in the samples from the dams (). These results demonstrate the vertical transmission of the genotoxic effect of the compounds tested. In conclusion, assessing MNEs in breastfeeding neonate rats to assess DNA damage may be a useful approach for identifying genotoxic compounds and/or cytotoxic effects. This strategy could help in screening for therapeutic approaches that are genotoxic during the lactation stage and these assessments might also be helpful for developing preventive strategies to counteract harmful effects. Belinda C. Gómez-Meda, Luis R. Bañales-Martínez, Ana L. Zamora-Perez, María de Lourdes Lemus-Varela, Xóchitl Trujillo, María G. Sánchez-Parada, Blanca M. Torres-Mendoza, Juan Armendáriz-Borunda, and Guillermo M. Zúñiga-González Copyright © 2016 Belinda C. Gómez-Meda et al. All rights reserved. Reproductive Toxicities Caused by Swainsonine from Locoweed in Mice Thu, 24 Nov 2016 14:10:51 +0000 Swainsonine is the primary toxin in locoweeds. It causes intention tremors, reproductive dysfunction, emaciation, and death. The objective of the present study was to evaluate the potential reproductive and developmental toxicities caused by swainsonine in mice. The treatment groups consisting of three generations of mice were given a range of concentrations of swainsonine by intraperitoneal injection (2.50 mg/kg body weight (BW), 1.20 mg/kg BW, 0.60 mg/kg BW, and 0 mg/kg BW). The 0 mg/kg BW group exhibited significantly fewer estrous cycles and an increased number of estrous ones compared to the 2.50 mg/kg BW, 1.20 mg/kg BW, and 0.60 mg/kg BW groups . All three generations of mice treated with swainsonine had significantly higher spleen, liver, and kidney indices and significantly lower body weights compared to the 0 mg/kg BW group . For the first and second generations of treatment group, the copulation indices and the numbers of live pups on postnatal days (PND) 0, 4, and 15 were significantly decreased compared to those of the 0 mg/kg BW group . The fertility and gestation indices of the treatment group of the first generation were significantly increased compared to the 2.50 mg/kg BW, 1.20 mg/kg BW, and 0.60 mg/kg BW groups of the second generation . Cumulatively, these results indicate that swainsonine may cause reproductive and developmental toxicities in mice in both parents and offspring. Chenchen Wu, Ke Feng, Dezhang Lu, Dujian Yan, Tiesuo Han, and Baoyu Zhao Copyright © 2016 Chenchen Wu et al. All rights reserved. Environmental Characteristics of Polybrominated Diphenyl Ethers in Marine System, with Emphasis on Marine Organisms and Sediments Thu, 24 Nov 2016 07:27:08 +0000 Polybrominated diphenyl ethers (PBDEs), due to their widespread usage as flame retardants and their lipophilicity and persistence, have become ubiquitous in the environment. It is urgent to understand the environmental characteristics of PBDEs in marine system, but they have attracted little attention. We summarize the available data and analyze the regional distributions, controlling factors, and congener patterns of PBDEs in marine and associated environmental matrixes worldwide. Based on meta-analysis, after separating the estuarial sites from the marine sites, ignoring the extraordinary sample sites such as those located just near the point source, the PBDE concentration levels are still in the same order of magnitude from global scale. Despite Principal Component Analysis, the congener patterns of sediments are predominant with the heavy brominated congeners (BDE-209 contributing over 75% to the total load) while the biota abound with the light ones (BDE-47, BDE-99, and BDE-100 taking about 80%). The ratio between BDE-99 and BDE-100 for the lower trophic-level species often turns to be greater than 1, while for those higher species the ratio may be below 1, and some species feed mainly on the crustaceans and zooplankton seems to have a higher ratio value. The data of the PBDEs in marine system are currently limited; thus, data gaps are identified as well. Ying Zhang, Weiliang Wang, Jinming Song, Zongming Ren, Huamao Yuan, Huijun Yan, Jinpeng Zhang, Zhen Pei, and Zhipeng He Copyright © 2016 Ying Zhang et al. All rights reserved. Integrative Characterization of Toxic Response of Zebra Fish (Danio rerio) to Deltamethrin Based on AChE Activity and Behavior Strength Wed, 23 Nov 2016 14:01:08 +0000 In order to characterize the toxic response of zebra fish (Danio rerio) to Deltamethrin (DM), behavior strength (BS) and muscle AChE activity of zebra fish were investigated. The results showed that the average values of both BS and AChE activity showed a similarly decreased tendency as DM concentration increased, which confirmed the dose-effect relationship, and high and low levels of AChE and BS partly matched low and high levels of exposure concentrations in self-organizing map. These indicated that AChE and BS had slight different aspects of toxicity although overall trend was similar. Behavior activity suggested a possibility of reviving circadian rhythm in test organisms after exposure to the chemical in lower concentration (0.1 TU). This type of rhythm disappeared in higher concentrations (1.0 TU and 2.0 TU). Time series trend analysis of BS and AChE showed an evident time delayed effect of AChE, and a 2 h AChE inhibition delay with higher correlation coefficients () in different treatments was observed. It was confirmed that muscle AChE inhibition of zebra fish is a factor for swimming behavior change, though there was a 2 h delay, and other factors should be investigated to illustrate the detailed behavior response mechanism. Qing Ren, Tingting Zhang, Shangge Li, Zongming Ren, Meiyi Yang, Hongwei Pan, Shiguo Xu, Li Qi, and Tae-Soo Chon Copyright © 2016 Qing Ren et al. All rights reserved. Effect of Salinity and Alkalinity on Luciobarbus capito Gill Na+/K+-ATPase Enzyme Activity, Plasma Ion Concentration, and Osmotic Pressure Thu, 17 Nov 2016 13:49:13 +0000 We evaluated the individual and combined effects of salinity and alkalinity on gill Na+/K+-ATPase enzyme activity, plasma ion concentration, and osmotic pressure in Luciobarbus capito. Increasing salinity concentrations (5, 8, 11, and 14 g/L) were associated with an initial increase and then decrease in L. capito gill Na+/K+-ATPase activity. Activity was affected by the difference between internal and external Na+ ion concentrations and osmotic pressure (). Both plasma ion (Na+, K+, and Cl−) concentration and osmotic pressure increased significantly (). An increase in alkalinity (15, 30, 45, and 60 mM) caused a significant increase in plasma K+ and urea nitrogen concentrations () but had no effect on either plasma osmotic pressure or gill filament ATPase activity. In the two-factor experiment, the saline-alkaline interaction caused a significant increase in plasma ion (Na+, Cl−, and urea nitrogen) and osmotic pressure (). Variance analysis revealed that salinity, alkalinity, and their interaction significantly affected osmotic pressure, with salinity being most affected, followed by alkalinity, and their interaction. Gill filament ATPase activity increased at first and then decreased; peak values were observed in the orthogonal experiment group at a salinity of 8 g/L and alkalinity of 30 mM. Longwu Geng, Guangxiang Tong, Haifeng Jiang, and Wei Xu Copyright © 2016 Longwu Geng et al. All rights reserved. Migration of BTEX and Biodegradation in Shallow Underground Water through Fuel Leak Simulation Tue, 01 Nov 2016 09:51:42 +0000 To provide more reasonable references for remedying underground water, fuel leak was simulated by establishing an experimental model of a porous-aquifer sand tank with the same size as that of the actual tank and by monitoring the underground water. In the tank, traditional gasoline and ethyl alcohol gasoline were poured. This study was conducted to achieve better understanding of the migration and distribution of benzene, toluene, ethyl benzene, and xylene (BTEX), which are major pollutants in the underground water. Experimental results showed that, compared with conventional gasoline, the content peak of BTEX in the mixture of ethyl alcohol gasoline appeared later; BTEX migrated along the water flow direction horizontally and presented different pollution halos; BTEX also exhibited the highest content level at 45 cm depth; however, its content declined at the 30 and 15 cm depths vertically because of the vertical dispersion effect; the rise of underground water level increased the BTEX content, and the attenuation of BTEX content in underground water was related to the biodegradation in the sand tank, which mainly included biodegradation with oxygen, nitrate, and sulfate. Yaping Cheng, Yudao Chen, Yaping Jiang, Lingzhi Jiang, Liqun Sun, Liuyue Li, and Junyu Huang Copyright © 2016 Yaping Cheng et al. All rights reserved. Developmental Toxicity of Carbon Quantum Dots to the Embryos/Larvae of Rare Minnow (Gobiocypris rarus) Mon, 31 Oct 2016 06:37:38 +0000 The toxic effects of CDs on rare minnow (Gobiocypris rarus) embryos at different developmental stages were investigated. The results showed that rare minnow embryos had decreased spontaneous movements, body length, increased heart rate, pericardial edema, yolk sac edema, tail/spinal curvature, various morphological malformations, and decreased hatching rate. Biochemical analysis showed the CDs exposure significantly inhibited the activity of Na+/K+-ATPase and Ca2+-ATPase and increased the MDA contents and the activity of SOD, CAT, and GPX. Further examination suggested that the CDs exposure induced serious embryonic cellular DNA damage. Moreover, the CDs exposure induced upregulation of development related genes (Wnt8a and Mstn) along with the downregulation of Vezf1. Overall, the present study revealed that the CDs exposure has significant development toxicity on rare minnow embryos/larvae. Mechanistically, this toxicity might result from the pressure of induced oxidative stress coordinate with the dysregulated development related gene expression mediated by the CDs exposure. Yuan-Yuan Xiao, Li Liu, Yao Chen, Yu-Lian Zeng, Ming-Zhi Liu, and Li Jin Copyright © 2016 Yuan-Yuan Xiao et al. All rights reserved. Spatiotemporal Distribution and Assemblages of Fishes below the Lowermost Dam in Protected Reach in the Yangtze River Main Stream: Implications for River Management Mon, 24 Oct 2016 12:35:07 +0000 Now more and more ecologists concern about the impacts of dam construction on fish. However, studies of fishes downstream Gezhouba Dam were rarely reported except Chinese sturgeon (Acipenser sinensis Gray). In this study, catch investigations and five hydroacoustic detections were completed from 2015 to 2016 to understand the distribution, size, and categories of fishes and their relationship with the environmental factors below Gezhouba Dam in protected reach in the Yangtze River main stream. Results showed significant differences in fish distribution and TS (target strength) between wet and flood seasons. Mean TS in five hydroacoustic detections were −59.98 dB, −54.70 dB, −56.16 dB, −57.90 dB, and −59.17 dB, respectively, and dominant fish species are Coreius guichenoti (Bleeker), Siniperca chuatsi (Basilewsky), and Pelteobagrus vachelli (Richardson). In the longitudinal direction, fish preferred to stay in some specific sections like reaches 2, 4, 7, 8, 11, and 16. Since hydrology factors change greatly in different seasons, environmental characteristics vary along the reaches, and human activities play an important role in the fish behavior, it is concluded that great cross-season changes in hydrology lead to the differences in TS and fish assemblages and that geography characteristics, especially channel geography, together with human activities influence fish longitudinal distribution. This finding provides basic knowledge of spatiotemporal distribution and assemblages of fishes in the extended reaches downstream Gezhouba Dam. In addition, it offers implications for river management. It could also serve as reference of future research on fish habitat. Junyi Li, Hui Zhang, Danqing Lin, Jinming Wu, Chengyou Wang, Xuan Xie, and Qiwei Wei Copyright © 2016 Junyi Li et al. All rights reserved. Isolation, Identification, and Optimization of Culture Conditions of a Bioflocculant-Producing Bacterium Bacillus megaterium SP1 and Its Application in Aquaculture Wastewater Treatment Thu, 20 Oct 2016 14:30:56 +0000 A bioflocculant-producing bacterium, Bacillus megaterium SP1, was isolated from biofloc in pond water and identified by using both 16S rDNA sequencing analysis and a Biolog GEN III MicroStation System. The optimal carbon and nitrogen sources for Bacillus megaterium SP1 were 20 g L−1 of glucose and 0.5 g L−1 of beef extract at 30°C and pH 7. The bioflocculant produced by strain SP1 under optimal culture conditions was applied into aquaculture wastewater treatment. The removal rates of chemical oxygen demand (COD), total ammonia nitrogen (TAN), and suspended solids (SS) in aquaculture wastewater reached 64, 63.61, and 83.8%, respectively. The volume of biofloc (FV) increased from 4.93 to 25.97 mL L−1. The addition of Bacillus megaterium SP1 in aquaculture wastewater could effectively improve aquaculture water quality, promote the formation of biofloc, and then form an efficient and healthy aquaculture model based on biofloc technology. Liang Luo, Zhigang Zhao, Xiaoli Huang, Xue Du, Chang’an Wang, Jinnan Li, Liansheng Wang, and Qiyou Xu Copyright © 2016 Liang Luo et al. All rights reserved. Exposure to p,p′-DDE Induces Morphological Changes and Activation of the PKCα-p38-C/EBPβ Pathway in Human Promyelocytic HL-60 Cells Wed, 19 Oct 2016 12:12:44 +0000 Dichlorodiphenyldichloroethylene (p,p′-DDE), the most persistent metabolite of dichlorodiphenyltrichloroethane (DDT), is still present in the human population. Both are present in the bone marrow of patients with bone marrow disorders, but thus far there are no studies that assess the capability of p,p′-DDE to affect myeloid cells. The aim of this study was to determine the effect of p,p′-DDE on promyelocytic cell differentiation and intracellular pathways related to this event. p,p′-DDE induced morphological changes compatible with promyelocytic differentiation in a concentration-dependent manner. The p,p′-DDE effect on , C/EBPβ protein levels, PKCα and p38 activation, and the role of oxidative stress or PLA2 was assayed. Exposure to 1.9 μg/mL of p,p′-DDE increased , PKCα, p38, and C/EBPβ protein levels; the increase of nuclear C/EBPβ protein was dependent on p38. PKCα phosphorylation was dependent on PLA2 and p,p′-DDE-induced oxidative stress. p38 phosphorylation induced by p,p′-DDE was dependent on PLA2, PKC activation, and oxidative stress. These effects of p,p′-DDE at concentrations found in human bone marrow may induce alterations in immature myeloid cells and could affect their cellular homeostasis. In order to establish the risk from exposure to p,p′-DDE on the development of bone marrow disorders in humans, these effects deserve further study. Nallely A. Torres-Avilés, Damaris Albores-García, Ana L. Luna, Monica Moreno-Galván, Mariana Salgado-Bustamante, Diana Patricia Portales-Pérez, and Emma S. Calderón-Aranda Copyright © 2016 Nallely A. Torres-Avilés et al. All rights reserved. FBS or BSA Inhibits EGCG Induced Cell Death through Covalent Binding and the Reduction of Intracellular ROS Production Tue, 18 Oct 2016 06:38:28 +0000 Previously we have shown that ()-epigallocatechin gallate (EGCG) can induce nonapoptotic cell death in human hepatoma HepG2 cells only under serum-free condition. However, the underlying mechanism for serum in determining the cell fate remains to be answered. The effects of fetal bovine serum (FBS) and its major component bovine serum albumin (BSA) on EGCG-induced cell death were investigated in this study. It was found that BSA, just like FBS, can protect cells from EGCG-induced cell death in a dose-dependent manner. Detailed analysis revealed that both FBS and BSA inhibited generation of ROS to protect against toxicity of EGCG. Furthermore, EGCG was shown to bind to certain cellular proteins including caspase-3, PARP, and α-tubulin, but not LC3 nor β-actin, which formed EGCG-protein complexes that were inseparable by SDS-gel. On the other hand, addition of FBS or BSA to culture medium can block the binding of EGCG to these proteins. In silico docking analysis results suggested that BSA had a stronger affinity to EGCG than the other proteins. Taken together, these data indicated that the protective effect of FBS and BSA against EGCG-induced cell death could be due to (1) the decreased generation of ROS and (2) the competitive binding of BSA to EGCG. Yin Zhang, Yu-Ying Xu, Wen-Jie Sun, Mo-Han Zhang, Yi-Fan Zheng, Han-Ming Shen, Jun Yang, and Xin-Qiang Zhu Copyright © 2016 Yin Zhang et al. All rights reserved. Microcystin Biosynthesis and mcyA Expression in Geographically Distinct Microcystis Strains under Different Nitrogen, Phosphorus, and Boron Regimes Mon, 10 Oct 2016 09:17:13 +0000 Roles of nutrients and other environmental variables in development of cyanobacterial bloom and its toxicity are complex and not well understood. We have monitored the photoautotrophic growth, total microcystin concentration, and microcystins synthetase gene (mcyA) expression in lab-grown strains of Microcystis NIES 843 (reference strain), KW (Wangsong Reservoir, South Korea), and Durgakund (Varanasi, India) under different nutrient regimes (nitrogen, phosphorus, and boron). Higher level of nitrogen and boron resulted in increased growth (avg. 5 and 6.5 Chl mg/L, resp.), total microcystin concentrations (avg. 1.185 and 7.153 mg/L, resp.), and mcyA transcript but its expression was not directly correlated with total microcystin concentrations in the target strains. Interestingly, Durgakund strain had much lower microcystin content and lacked microcystin-YR variant over NIES 843 and KW. It is inferred that microcystin concentration and its variants are strain specific. We have also examined the heterotrophic bacteria associated with cyanobacterial bloom in Durgakund Pond and Wangsong Reservoir which were found to be enriched in Alpha-, Beta-, and Gammaproteobacteria and that could influence the bloom dynamics. Ankita Srivastava, So-Ra Ko, Chi-Yong Ahn, Hee-Mock Oh, Alok Kumar Ravi, and Ravi Kumar Asthana Copyright © 2016 Ankita Srivastava et al. All rights reserved. Advances in Understanding How Heavy Metal Pollution Triggers Gastric Cancer Mon, 10 Oct 2016 08:56:49 +0000 With the development of industrialization and urbanization, heavy metals contamination has become a major environmental problem. Numerous investigations have revealed an association between heavy metal exposure and the incidence and mortality of gastric cancer. The mechanisms of heavy metals (lead, cadmium, mercury, chromium, and arsenic) contamination leading to gastric cancer are concluded in this review. There are four main potential mechanisms: (1) Heavy metals disrupt the gastric mucosal barrier by decreasing mucosal thickness, mucus content, and basal acid output, thereby affecting the function of E-cadherin and inducing reactive oxygen species (ROS) damage. (2) Heavy metals directly or indirectly induce ROS generation and cause gastric mucosal and DNA lesions, which subsequently alter gene regulation, signal transduction, and cell growth, ultimately leading to carcinogenesis. Exposure to heavy metals also enhances gastric cancer cell invasion and metastasis. (3) Heavy metals inhibit DNA damage repair or cause inefficient lesion repair. (4) Heavy metals may induce other gene abnormalities. In addition, heavy metals can induce the expression of proinflammatory chemokine interleukin-8 (IL-8) and microRNAs, which promotes tumorigenesis. The present review is an effort to underline the human health problem caused by heavy metal with recent development in order to garner a broader perspective. Wenzhen Yuan, Ning Yang, and Xiangkai Li Copyright © 2016 Wenzhen Yuan et al. All rights reserved. Silver Nanoparticles Affect Functional Bioenergetic Traits in the Invasive Red Sea Mussel Brachidontes pharaonis Wed, 05 Oct 2016 14:07:06 +0000 We investigated the functional trait responses to 5 nm metallic silver nanoparticle (AgNPs) exposure in the Lessepsian-entry bivalve B. pharaonis. Respiration rate (oxygen consumption), heartbeat rate, and absorption efficiency were evaluated across an 8-day exposure period in mesocosmal conditions. Basal reference values from not-exposed specimens were statistically compared with those obtained from animals treated with three sublethal nanoparticle concentrations (2 μg L−1, 20 μg L−1, and 40 μg L−1). Our data showed statistically significant effects on the average respiration rate of B. pharaonis. Moreover, complex nonlinear dynamics were observed as a function of the concentration level and time. Heartbeat rates largely increased with no acclimation in animals exposed to the two highest levels with similar temporal dynamics. Eventually, a decreasing trend for absorption efficiency might indicate energetic constraints. In general, these data support the possible impact of engineered nanomaterials in marine environments and support the relevance of functional trait assessment in present and future ecotoxicological studies. Ilenia Saggese, Gianluca Sarà, and Francesco Dondero Copyright © 2016 Ilenia Saggese et al. All rights reserved. Human Elimination of Organochlorine Pesticides: Blood, Urine, and Sweat Study Wed, 05 Oct 2016 08:05:14 +0000 Background. Many individuals have been exposed to organochlorinated pesticides (OCPs) through food, water, air, dermal exposure, and/or vertical transmission. Due to enterohepatic reabsorption and affinity to adipose tissue, OCPs are not efficiently eliminated from the human body and may accrue in tissues. Many epidemiological studies demonstrate significant exposure-disease relationships suggesting OCPs can alter metabolic function and potentially lead to illness. There is limited study of interventions to facilitate OCP elimination from the human body. This study explored the efficacy of induced perspiration as a means to eliminate OCPs. Methods. Blood, urine, and sweat (BUS) were collected from 20 individuals. Analysis of 23 OCPs was performed using dual-column gas chromatography with electron-capture detectors. Results. Various OCPs and metabolites, including DDT, DDE, methoxychlor, endrin, and endosulfan sulfate, were excreted into perspiration. Generally, sweat samples showed more frequent OCP detection than serum or urine analysis. Many OCPs were not readily detected in blood testing while still being excreted and identified in sweat. No direct correlation was found among OCP concentrations in the blood, urine, or sweat compartments. Conclusions. Sweat analysis may be useful in detecting some accrued OCPs not found in regular serum testing. Induced perspiration may be a viable clinical tool for eliminating some OCPs. Stephen J. Genuis, Kevin Lane, and Detlef Birkholz Copyright © 2016 Stephen J. Genuis et al. All rights reserved. Environmental and Ecological Risk Assessment of Trace Metal Contamination in Mangrove Ecosystems: A Case from Zhangjiangkou Mangrove National Nature Reserve, China Tue, 04 Oct 2016 09:15:20 +0000 Zhangjiangkou Mangrove National Nature Reserve is a subtropical wetland ecosystem in southeast coast of China, which is of dense population and rapid development. The concentrations, sources, and pollution assessment of trace metals (Cu, Cd, Pb, Cr, Zn, As, and Hg) in surface sediment from 29 sites and the biota specimen were investigated for better ecological risk assessment and environmental management. The ranges of trace metals in mg/kg sediment were as follows: Cu (10.79–26.66), Cd (0.03–0.19), Pb (36.71–59.86), Cr (9.67–134.51), Zn (119.69–157.84), As (15.65–31.60), and Hg (0.00–0.08). The sequences of the bioaccumulation of studied metals are Zn > Cu > As > Cr > Pb > Cd > Hg with few exceptions. Cluster analysis and principal component analysis revealed that the trace metals in the studied area mainly derived from anthropogenic activities, such as industrial effluents, agricultural waste, and domestic sewage. Pollution load index and geoaccumulation index were calculated for trace metals in surface sediments, which indicated unpolluted status in general except Pb, Cr, and As. Jun Wang, Huihong Du, Ye Xu, Kai Chen, Junhua Liang, Hongwei Ke, Sha-Yen Cheng, Mengyang Liu, Hengxiang Deng, Tong He, Wenqing Wang, and Minggang Cai Copyright © 2016 Jun Wang et al. All rights reserved. Sequestration and Distribution Characteristics of Cd(II) by Microcystis aeruginosa and Its Role in Colony Formation Thu, 29 Sep 2016 12:10:36 +0000 To investigate the sequestration and distribution characteristics of Cd(II) by Microcystis aeruginosa and its role in Microcystis colony formation, M. aeruginosa was exposed to six different Cd(II) concentrations for 10 days. Cd(II) exposure caused hormesis in the growth of M. aeruginosa. Low concentrations of Cd(II) significantly induced formation of small Microcystis colonies () and increased the intracellular polysaccharide (IPS) and bound extracellular polysaccharide (bEPS) contents of M. aeruginosa significantly (). There was a linear relationship between the amount of Cd(II) sequestrated by algal cells and the amount added to cultures in the rapid adsorption process that occurred during the first 5 min of exposure. After 10 d, M. aeruginosa sequestrated nearly 80% of 0.2 mg L−1 added Cd(II), while >93% of Cd(II) was sequestrated in the groups with lower added concentrations of Cd(II). More than 80% of the sequestrated Cd(II) was bioadsorbed by bEPS. The Pearson correlation coefficients of exterior and interior factors related to colony formation of M. aeruginosa revealed that Cd(II) could stimulate the production of IPS and bEPS via increasing Cd(II) bioaccumulation and bioadsorption. Increased levels of cross-linking between Cd(II) and bEPS stimulated algal cell aggregation, which eventually promoted the formation of Microcystis colonies. Xiangdong Bi, Ran Yan, Fenxiang Li, Wei Dai, Kewei Jiao, Qixing Zhou, and Qi Liu Copyright © 2016 Xiangdong Bi et al. All rights reserved. Optimization of a Coastal Environmental Monitoring Network Based on the Kriging Method: A Case Study of Quanzhou Bay, China Thu, 29 Sep 2016 08:59:17 +0000 Environmental monitoring is fundamental in assessing environmental quality and to fulfill protection and management measures with permit conditions. However, coastal environmental monitoring work faces many problems and challenges, including the fact that monitoring information cannot be linked up with evaluation, monitoring data cannot well reflect the current coastal environmental condition, and monitoring activities are limited by cost constraints. For these reasons, protection and management measures cannot be developed and implemented well by policy makers who intend to solve this issue. In this paper, Quanzhou Bay in southeastern China was selected as a case study; and the Kriging method and a geographic information system were employed to evaluate and optimize the existing monitoring network in a semienclosed bay. This study used coastal environmental monitoring data from 15 sites (including COD, DIN, and PO4-P) to adequately analyze the water quality from 2009 to 2012 by applying the Trophic State Index. The monitoring network in Quanzhou Bay was evaluated and optimized, with the number of sites increased from 15 to 24, and the monitoring precision improved by 32.9%. The results demonstrated that the proposed advanced monitoring network optimization was appropriate for environmental monitoring in Quanzhou Bay. It might provide technical support for coastal management and pollutant reduction in similar areas. Kai Chen, Minjie Ni, Minggang Cai, Jun Wang, Dongren Huang, Huorong Chen, Xiao Wang, and Mengyang Liu Copyright © 2016 Kai Chen et al. All rights reserved. The Effect of Kanamycin and Tetracycline on Growth and Photosynthetic Activity of Two Chlorophyte Algae Sun, 25 Sep 2016 07:12:01 +0000 Antibiotics are routinely used in microalgae culture screening, stock culture maintenance, and genetic transformation. By studying the effect of antibiotics on microalgae growth, we can estimate the least value to inhibit growth of undesired pathogens in algal culture. We studied the effect of kanamycin and tetracycline on the growth and photosynthetic activity of two chlorophyte microalgae, Dictyosphaerium pulchellum and Micractinium pusillum. We measured CFU mL−1 on agar plates, optical density, fluorescence yields, and photosynthetic inhibition. Our results showed a significant effect of kan and tet on the tested microalgae species except tet, which showed a minor effect on M. pusillum. Both antibiotics are believed to interact with the protein synthesis machinery; hence, the inhibitory effect of the tested antibiotics was further confirmed by isolation and quantification of the whole cell protein. A significant reduction in protein quantity was observed at concentrations more than 5 mg L−1, except M. pusillum, which showed only a slight reduction in protein quantity even at the maximum tested concentration of tet (30 mg L−1). This study can further aid in aquaculture industry, for the maintenance of the microalgae stock cultures and it can also help the microalgae genetic engineers in the construction of molecular markers. Khawaja Muhammad Imran Bashir and Man-Gi Cho Copyright © 2016 Khawaja Muhammad Imran Bashir and Man-Gi Cho. All rights reserved. Advances in Engineered Liver Models for Investigating Drug-Induced Liver Injury Tue, 20 Sep 2016 16:36:17 +0000 Drug-induced liver injury (DILI) is a major cause of drug attrition. Testing drugs on human liver models is essential to mitigate the risk of clinical DILI since animal studies do not always suffice due to species-specific differences in liver pathways. While primary human hepatocytes (PHHs) can be cultured on extracellular matrix proteins, a rapid decline in functions leads to low sensitivity (<50%) in DILI prediction. Semiconductor-driven engineering tools now allow precise control over the hepatocyte microenvironment to enhance and stabilize phenotypic functions. The latest platforms coculture PHHs with stromal cells to achieve hepatic stability and enable crosstalk between the various liver cell types towards capturing complex cellular mechanisms in DILI. The recent introduction of induced pluripotent stem cell-derived human hepatocyte-like cells can potentially allow a better understanding of interindividual differences in idiosyncratic DILI. Liver models are also being coupled to other tissue models via microfluidic perfusion to study the intertissue crosstalk upon drug exposure as in a live organism. Here, we review the major advances being made in the engineering of liver models and readouts as they pertain to DILI investigations. We anticipate that engineered human liver models will reduce drug attrition, animal usage, and cases of DILI in humans. Christine Lin and Salman R. Khetani Copyright © 2016 Christine Lin and Salman R. Khetani. All rights reserved. Key Challenges and Opportunities Associated with the Use of In Vitro Models to Detect Human DILI: Integrated Risk Assessment and Mitigation Plans Mon, 05 Sep 2016 07:48:31 +0000 Drug-induced liver injury (DILI) is a major cause of late-stage clinical drug attrition, market withdrawal, black-box warnings, and acute liver failure. Consequently, it has been an area of focus for toxicologists and clinicians for several decades. In spite of considerable efforts, limited improvements in DILI prediction have been made and efforts to improve existing preclinical models or develop new test systems remain a high priority. While prediction of intrinsic DILI has improved, identifying compounds with a risk for idiosyncratic DILI (iDILI) remains extremely challenging because of the lack of a clear mechanistic understanding and the multifactorial pathogenesis of idiosyncratic drug reactions. Well-defined clinical diagnostic criteria and risk factors are also missing. This paper summarizes key data interpretation challenges, practical considerations, model limitations, and the need for an integrated risk assessment. As demonstrated through selected initiatives to address other types of toxicities, opportunities exist however for improvement, especially through better concerted efforts at harmonization of current, emerging and novel in vitro systems or through the establishment of strategies for implementation of preclinical DILI models across the pharmaceutical industry. Perspectives on the incorporation of newer technologies and the value of precompetitive consortia to identify useful practices are also discussed. Franck A. Atienzar, Eric A. Blomme, Minjun Chen, Philip Hewitt, J. Gerry Kenna, Gilles Labbe, Frederic Moulin, Francois Pognan, Adrian B. Roth, Laura Suter-Dick, Okechukwu Ukairo, Richard J. Weaver, Yvonne Will, and Donna M. Dambach Copyright © 2016 Franck A. Atienzar et al. All rights reserved. Identifying the Types of Ion Channel-Targeted Conotoxins by Incorporating New Properties of Residues into Pseudo Amino Acid Composition Thu, 18 Aug 2016 16:35:36 +0000 Conotoxins are a kind of neurotoxin which can specifically interact with potassium, sodium type, and calcium channels. They have become potential drug candidates to treat diseases such as chronic pain, epilepsy, and cardiovascular diseases. Thus, correctly identifying the types of ion channel-targeted conotoxins will provide important clue to understand their function and find potential drugs. Based on this consideration, we developed a new computational method to rapidly and accurately predict the types of ion-targeted conotoxins. Three kinds of new properties of residues were proposed to use in pseudo amino acid composition to formulate conotoxins samples. The support vector machine was utilized as classifier. A feature selection technique based on F-score was used to optimize features. Jackknife cross-validated results showed that the overall accuracy of 94.6% was achieved, which is higher than other published results, demonstrating that the proposed method is superior to published methods. Hence the current method may play a complementary role to other existing methods for recognizing the types of ion-target conotoxins. Yun Wu, Yufei Zheng, and Hua Tang Copyright © 2016 Yun Wu et al. All rights reserved. Interaction Effects between Organochlorine Pesticides and Isoflavones In Vitro and In Vivo Mon, 15 Aug 2016 11:41:22 +0000 Organochlorine pesticides (OCPs) have caused increasing global concern due to their high toxicity, persistence, bioaccumulation, and significant adverse effects on human health. This study was to explore the interaction effects between OCPs and isoflavones. Six kinds of OCPs and 2 kinds of isoflavones-genistein and daidzein were included to study their effect on MCF-7 cells in vitro. Eighty-one female Sprague-Dawley rats were randomized to 9 groups according to factorial design to study the interaction effect between isoflavones and γ-HCH. Compared to organochlorine pesticides alone group, proliferation rate of MCF-7 cells was lower in 100 μmol/L genistein + organochlorine pesticides and 100 μmol/L daidzein + organochlorine pesticides group (). In vivo study showed that there are interaction effects on kidney weight and liver weight when treated with isoflavones and γ-HCH. The changes in uterine morphology and positive expression of ERα showed inhibition effects between isoflavones and γ-HCH. In conclusion, the data suggests that there are interactions between isoflavones and OCPs in vitro and in vivo. Yunbo Zhang, Jipeng Guo, Xiao Zhang, Jingjing Guo, Ming Zhang, Yang Yang, and Xiaolin Na Copyright © 2016 Yunbo Zhang et al. All rights reserved. Expression of miRNA-122 Induced by Liver Toxicants in Zebrafish Wed, 03 Aug 2016 08:48:08 +0000 MicroRNA-122 (miRNA-122), also known as liver-specific miRNA, has recently been shown to be a potent biomarker in response to liver injury in mammals. The objective of this study was to examine its expression in response to toxicant treatment and acute liver damage, using the zebrafish system as an alternative model organism. For the hepatotoxicity assay, larval zebrafish were arrayed in 24-well plates. Adult zebrafish were also tested and arrayed in 200 mL cages. Animals were exposed to liver toxicants (tamoxifen or acetaminophen) at various doses, and miRNA-122 expression levels were analyzed using qRT-PCR in dissected liver, brain, heart, and intestine, separately. Our results showed no significant changes in miRNA-122 expression level in tamoxifen-treated larvae; however, miRNA-122 expression was highly induced in tamoxifen-treated adults in a tissue-specific manner. In addition, we observed a histological change in adult liver (0.5 μM) and cell death in larval liver (5 μM) at different doses of tamoxifen. These results indicated that miRNA-122 may be utilized as a liver-specific biomarker for acute liver toxicity in zebrafish. Hyun-Sik Nam, Kyu-Seok Hwang, Yun-Mi Jeong, Jeong-Im Ryu, Tae-Young Choi, Myung-Ae Bae, Woo-Chan Son, Kwan-Hee You, Hwa-Young Son, and Cheol-Hee Kim Copyright © 2016 Hyun-Sik Nam et al. All rights reserved. A Gram-Negative Bacterial Secreted Protein Types Prediction Method Based on PSI-BLAST Profile Tue, 02 Aug 2016 09:52:32 +0000 Prediction of secreted protein types based solely on sequence data remains to be a challenging problem. In this study, we extract the long-range correlation information and linear correlation information from position-specific score matrix (PSSM). A total of 6800 features are extracted at 17 different gaps; then, 309 features are selected by a filter feature selection method based on the training set. To verify the performance of our method, jackknife and independent dataset tests are performed on the test set and the reported overall accuracies are 93.60% and 100%, respectively. Comparison of our results with the existing method shows that our method provides the favorable performance for secreted protein type prediction. Shuyan Ding and Shengli Zhang Copyright © 2016 Shuyan Ding and Shengli Zhang. All rights reserved. In Vivo and In Vitro Genotoxic and Epigenetic Effects of Two Types of Cola Beverages and Caffeine: A Multiassay Approach Mon, 04 Jul 2016 07:31:10 +0000 The aim of this work was to assess the biological and food safety of two different beverages: Classic Coca Cola™ (CCC) and Caffeine-Free Coca Cola (CFCC). To this end, we determined the genotoxicological and biological effects of different doses of lyophilised CCC and CFCC and Caffeine (CAF), the main distinctive constituent. Their toxic/antitoxic, genotoxic/antigenotoxic, and chronic toxicity (lifespan assay) effects were determined in vivo using the Drosophila model. Their cytotoxic activities were determined using the HL-60 in vitro cancer model. In addition, clastogenic DNA toxicity was measured using internucleosomal fragmentation and SCGE assays. Their epigenetic effects were assessed on the HL-60 methylation status using some repetitive elements. The experimental results showed a slight chemopreventive effect of the two cola beverages against HL-60 leukaemia cells, probably mediated by nonapoptotic mechanisms. Finally, CCC and CAF induced a global genome hypomethylation evaluated in LINE-1 and Alu M1 repetitive elements. Overall, we demonstrated for the first time the safety of this famous beverage in in vivo and in vitro models. Marcos Mateo-Fernández, Tania Merinas-Amo, Miguel Moreno-Millán, Ángeles Alonso-Moraga, and Sebastián Demyda-Peyrás Copyright © 2016 Marcos Mateo-Fernández et al. All rights reserved. Health Issues of Primary School Students Residing in Proximity of an Oil Terminal with Environmental Exposure to Volatile Organic Compounds Sun, 03 Jul 2016 07:48:51 +0000 Residential proximity to industrial sites has been associated with adverse effects on human health. Children are more susceptible to airborne environmental exposure because their immune and respiratory systems are still developing. This study aimed to investigate whether living close to an oil terminal in Genoa where there is higher VOCs exposure is associated with an increased rate of school absenteeism because of disease in primary school children. Five schools were chosen for the recruitment of children and students residing in the industrial site (A) were compared to those living in residential sites (B). Sixty-six of the 407 students involved in the project were also selected for VOC monitoring. Source apportionment was carried out by comparing profiles of VOCs; principal component analysis was performed to study the correlation between profiles, and Kriging interpolation model was used to extend profiles to all participants. The concentration means of total VOCs were significantly higher in the industrial areas compared to controls. Adjusting for potential confounders, children who lived in area A had a significantly higher risk of being absent from school due to sore throat, cough, and cold compared to controls. o-Xylene, which is dispersed during the industrial activity, showed clear evidence of a significant association with respiratory symptoms. Massimo Cipolla, Marco Bruzzone, Emanuele Stagnaro, Marcello Ceppi, Alberto Izzotti, Claudio Culotta, and Maria Teresa Piccardo Copyright © 2016 Massimo Cipolla et al. All rights reserved. Use of Mercury in Dental Silver Amalgam: An Occupational and Environmental Assessment Thu, 30 Jun 2016 11:55:39 +0000 The objective of this study was to assess the occupational exposure to mercury in dentistry and associated environmental emission in wastewater of Lahore, Pakistan. A total of ninety-eight blood samples were collected comprising 37 dentists, 31 dental assistants, and 30 controls. Results demonstrate that the dentistry personnel contained significantly higher mean concentration of mercury in their blood samples (dentists: 29.835 µg/L and dental assistants: 22.798 µg/L) compared to that of the controls (3.2769 µg/L). The mean concentration of mercury was found maximum in the blood samples of older age group (62.8 µg/L) in dentists and (44.3 µg/L) in dental assistants. The comparison of mercury concentration among dentists, dental assistants, and controls (pairing based on their ages) revealed that the concentration increased with the age and experience among the dentists and dental assistants. Moreover, the mercury concentration in all the studied dental wastewater samples, collected from twenty-two dental clinics, was found to be exceeding the recommended discharge limit of 0.01 mg/L. Therefore, we recommend that immediate steps must be taken to ensure appropriate preventive measures to avoid mercury vapors in order to prevent potential health hazards to dentistry personnel. Strong regulatory and administrative measures are needed to deal with mercury pollution on emergency basis. Nadia Jamil, Mujtaba Baqar, Samar Ilyas, Abdul Qadir, Muhammad Arslan, Muhammad Salman, Naveed Ahsan, and Hina Zahid Copyright © 2016 Nadia Jamil et al. All rights reserved. Contaminants of Emerging Concern: From the Detection to Their Effects on Human Health Mon, 20 Jun 2016 15:03:05 +0000 Fernando Barbosa Junior, Andres Campiglia, Bruno Rocha, and Daniel Cyr Copyright © 2016 Fernando Barbosa Junior et al. All rights reserved. Effects of Acute and Chronic Heavy Metal (Cu, Cd, and Zn) Exposure on Sea Cucumbers (Apostichopus japonicus) Sun, 12 Jun 2016 07:31:42 +0000 Acute and chronic toxicity tests were conducted with sea cucumber (Apostichopus japonicus) exposed to heavy metals. Acute toxicity values (96 h LC50) were 2.697, 0.133, and 1.574 mg L−1 for Zn, Cu, and Cd, respectively, and were ranked in order of toxicity: Cu > Cd > Zn. Under chronic metal exposure the specific growth rates of sea cucumbers decreased with the increase of metal concentration for all the three metals. After acute metal exposure, the oxygen consumption rate (OCR) decreased. The OCRs in all groups were significantly different than control () except in the group treated with 1.00 mg L−1 Zn (), where the increase of OCR was observed. The OCRs in groups chronically exposed to metals were significantly lower than that in the control group (). The activity of both pyruvate kinase (PK) and hexokinase (HK) in sea cucumbers followed: respiratory tree > muscle > intestine in natural sea water. After chronic Zn, Cu, and Cd exposure, the change pattern of HK and PK in respiratory tree, muscle, and intestine varied slightly. However, the activity of the enzyme showed a general trend of increase and then decrease and the higher the exposure concentration was, the earlier the highest point of enzyme activity was obtained. Li Li, Xiangli Tian, Xiao Yu, and Shuanglin Dong Copyright © 2016 Li Li et al. All rights reserved. Ultrastructural Mapping of the Zebrafish Gastrointestinal System as a Basis for Experimental Drug Studies Thu, 02 Jun 2016 13:30:49 +0000 Research in the field of gastroenterology is increasingly focused on the use of alternative nonrodent model organisms to provide new experimental tools to study chronic diseases. The zebrafish is a particularly valuable experimental platform to explore organ and cell structure-function relationships under relevant biological and pathobiological settings. This is due to its optical transparency and its close-to-human genetic makeup. To-date, the structure-function properties of the GIS of the zebrafish are relatively unexplored and limited to histology and fluorescent microscopy. Occasionally those studies include EM of a given subcellular process but lack the required full histological picture. In this work, we employed a novel combined biomolecular imaging approach in order to cross-correlate 3D ultrastructure over different length scales (optical-, X-ray micro-CT, and high-resolution EM). Our correlated imaging studies and subsequent data modelling provide to our knowledge the first detailed 3D picture of the zebrafish larvae GIS. Our results provide unequivocally a limit of confidence for studying various digestive disorders and drug delivery pathways in the zebrafish. Delfine Cheng, Gerald J. Shami, Marco Morsch, Roger S. Chung, and Filip Braet Copyright © 2016 Delfine Cheng et al. All rights reserved. Potential Role of Epigenetic Mechanism in Manganese Induced Neurotoxicity Thu, 26 May 2016 08:02:48 +0000 Manganese is a vital nutrient and is maintained at an optimal level (2.5–5 mg/day) in human body. Chronic exposure to manganese is associated with neurotoxicity and correlated with the development of various neurological disorders such as Parkinson’s disease. Oxidative stress mediated apoptotic cell death has been well established mechanism in manganese induced toxicity. Oxidative stress has a potential to alter the epigenetic mechanism of gene regulation. Epigenetic insight of manganese neurotoxicity in context of its correlation with the development of parkinsonism is poorly understood. Parkinson’s disease is characterized by the -synuclein aggregation in the form of Lewy bodies in neuronal cells. Recent findings illustrate that manganese can cause overexpression of -synuclein. -Synuclein acts epigenetically via interaction with histone proteins in regulating apoptosis. -Synuclein also causes global DNA hypomethylation through sequestration of DNA methyltransferase in cytoplasm. An individual genetic difference may also have an influence on epigenetic susceptibility to manganese neurotoxicity and the development of Parkinson’s disease. This review presents the current state of findings in relation to role of epigenetic mechanism in manganese induced neurotoxicity, with a special emphasis on the development of Parkinson’s disease. Prashant Tarale, Tapan Chakrabarti, Saravanadevi Sivanesan, Pravin Naoghare, Amit Bafana, and Kannan Krishnamurthi Copyright © 2016 Prashant Tarale et al. All rights reserved. Oxidative Stress and Liver Morphology in Experimental Cyclosporine A-Induced Hepatotoxicity Thu, 19 May 2016 17:11:17 +0000 Cyclosporine A is an immunosuppressive drug used after organ’s transplantation. The adverse effects on such organs as kidney or liver may limit its use. Oxidative stress is proposed as one of the mechanisms of organs injury. The study was designed to elucidate CsA-induced changes in liver function, morphology, oxidative stress parameters, and mitochondria in rat’s hepatocytes. Male Wistar rats were used: group A (control) receiving physiological saline, group B cyclosporine A in a dose of 15 mg/kg/day subcutaneously, and group C the CsA-vehicle (olive oil). On the 28th day rats were anesthetized. The following biochemical changes were observed in CsA-treated animals: increased levels of ALT, AST, and bilirubin in the serum, statistically significant changes in oxidative stress parameters, and lipid peroxidation products in the liver supernatants: MDA+4HAE, GSH, GSSG, caspase 3 activity, and ADP/ATP, NAD+/NADH, and NADP+/NADPH ratios. Microscopy of the liver revealed congestion, sinusoidal dilatation, and focal hepatocytes necrosis with mononuclear cell infiltration. Electron microscope revealed marked mitochondrial damage. Biochemical studies indicated that CsA treatment impairs liver function and triggers oxidative stress and redox imbalance in rats hepatocytes. Changes of oxidative stress markers parallel with mitochondrial damage suggest that these mechanisms play a crucial role in the course of CsA hepatotoxicity. Agnieszka Korolczuk, Kinga Caban, Magdalena Amarowicz, Grażyna Czechowska, and Joanna Irla-Miduch Copyright © 2016 Agnieszka Korolczuk et al. All rights reserved. Induction of Poly(ADP-ribose) Polymerase in Mouse Bone Marrow Stromal Cells Exposed to 900 MHz Radiofrequency Fields: Preliminary Observations Thu, 14 Apr 2016 06:09:31 +0000 Background. Several investigators have reported increased levels of poly(ADP-ribose) polymerase-1 (PARP-1), a nuclear enzyme which plays an important role in the repair of damaged DNA, in cells exposed to extremely low dose ionizing radiation which does not cause measurable DNA damage. Objective. To examine whether exposure of the cells to nonionizing radiofrequency fields (RF) is capable of increasing messenger RNA of PARP-1 and its protein levels in mouse bone marrow stromal cells (BMSCs). Methods. BMSCs were exposed to 900 MHz RF at 120 μW/cm2 power intensity for 3 hours/day for 5 days. PARP-1 mRNA and its protein levels were examined at 0, 0.5, 1, 2, 4, 6, 8, and 10 hours after exposure using RT-PCR and Western blot analyses. Sham-exposed (SH) cells and those exposed to ionizing radiation were used as unexposed and positive control cells. Results. BMSCs exposed to RF showed significantly increased expression of PARP-1 mRNA and its protein levels after exposure to RF while such changes were not observed in SH-exposed cells. Conclusion. Nonionizing RF exposure is capable of inducing PARP-1. Qina He, Yulong Sun, Lin Zong, Jian Tong, and Yi Cao Copyright © 2016 Qina He et al. All rights reserved. Comparison of the Effects of Dibutyl and Monobutyl Phthalates on the Steroidogenesis of Rat Immature Leydig Cells Mon, 11 Apr 2016 14:11:01 +0000 Dibutyl phthalate (DBP) is a widely used synthetic phthalic diester and monobutyl phthalate (MBP) is its main metabolite. DBP can be released into the environment and potentially disrupting mammalian male reproductive endocrine system. However, the potencies of DBP and MBP to inhibit Leydig cell steroidogenesis and their possible mechanisms are not clear. Immature Leydig cells isolated from rats were cultured with 0.05–50 M DBP or MBP for 3 h in combination with testosterone synthesis regulator or intermediate. The concentrations of 5-androstanediol and testosterone in the media were measured, and the mRNA levels of the androgen biosynthetic genes were detected by qPCR. The direct actions of DBP or MBP on CYP11A1, CYP17A1, SRD5A1, and AKR1C14 activities were measured. MBP inhibited androgen production by the immature Leydig cell at as low as 50 nM, while 50 M was required for DBP to suppress its androgen production. MBP mainly downregulated Cyp11a1 and Hsd3b1 expression levels at 50 nM. However, 50 M DBP downregulated Star, Hsd3b1, and Hsd17b3 expression levels and directly inhibited CYP11A1 and CYP17A1 activities. In conclusion, DBP is metabolized to more potent inhibitor MBP that downregulated the expression levels of some androgen biosynthetic enzymes. Linxi Li, Xiaomin Chen, Guoxin Hu, Sicong Wang, Renai Xu, Qiqi Zhu, Xiaoheng Li, Mingcang Wang, Qing-Quan Lian, and Ren-Shan Ge Copyright © 2016 Linxi Li et al. All rights reserved. Risks of Mycotoxins from Mycoinsecticides to Humans Sun, 10 Apr 2016 16:35:15 +0000 There are more than thirty mycotoxins produced by fungal entomopathogens. Totally, they belong to two classes, NRP and PK mycotoxins. Most of mycotoxins have not been paid sufficient attention yet. Generally, mycotoxins do not exist in mycoinsecticide and might not be released to environments unless entomogenous fungus proliferates and produces mycotoxins in host insects or probably in plants. Some mycotoxins, destruxins as an example, are decomposed in host insects before they, with the insect’s cadavers together, are released to environments. Many species of fungal entomopathogens have the endophytic characteristics. But we do not know if fungal entomopathogens produce mycotoxins in plants and release them to environments. On the contrary, the same mycotoxins produced by phytopathogens such as Fusarium spp. and Aspergillus spp. have been paid enough concerns. In conclusion, mycotoxins from mycoinsecticides have limited ways to enter environments. The risks of mycotoxins from mycoinsecticides contaminating foods are controllable. Qiongbo Hu, Fuxia Li, and Yuping Zhang Copyright © 2016 Qiongbo Hu et al. All rights reserved. Study of Valproic Acid-Enhanced Hepatocyte Steatosis Mon, 29 Feb 2016 06:48:55 +0000 Valproic acid (VPA) is one of the most widely used antiepilepsy drugs. However, several side effects, including weight gain and fatty liver, have been reported in patients following VPA treatment. In this study, we explored the molecular mechanisms of VPA-induced hepatic steatosis using FL83B cell line-based in vitro model. Using fluorescent lipid staining technique, we found that VPA enhanced oleic acid- (OLA-) induced lipid accumulation in a dose-dependent manner in hepatocytes; this may be due to upregulated lipid uptake, triacylglycerol (TAG) synthesis, and lipid droplet formation. Real-time PCR results showed that, following VPA treatment, the expression levels of genes encoding cluster of differentiation 36 (Cd36), low-density lipoprotein receptor-related protein 1 (Lrp1), diacylglycerol acyltransferase 2 (Dgat2), and perilipin 2 (Plin2) were increased, that of carnitine palmitoyltransferase I a (Cpt1a) was not affected, and those of acetyl-Co A carboxylase α (Acca) and fatty acid synthase (Fasn) were decreased. Furthermore, using immunofluorescence staining and flow cytometry analyses, we found that VPA also induced peroxisome proliferator-activated receptor γ (PPARγ) nuclear translocation and increased levels of cell-surface CD36. Based on these results, we propose that VPA may enhance OLA-induced hepatocyte steatosis through the upregulation of PPARγ- and CD36-dependent lipid uptake, TAG synthesis, and lipid droplet formation. Renin Chang, Mei-Chia Chou, Li-Ying Hung, Mu-En Wang, Meng-Chieh Hsu, and Chih-Hsien Chiu Copyright © 2016 Renin Chang et al. All rights reserved. Nanosilica and Polyacrylate/Nanosilica: A Comparative Study of Acute Toxicity Mon, 15 Feb 2016 07:19:55 +0000 We compared the acute toxicity of nanosilica and polyacrylate/nanosilica instillation in Wistar rats (). Exposure to nanosilica and polyacrylate/nanosilica showed a 30% mortality rate. When compared with saline-treated rats, animals in both exposure groups exhibited a significant reduction of PO2 () at both 24 and 72 hr. after exposure. Both exposure groups exhibited a significant reduction of neutrophils in arterial blood compared to saline controls () 24 hr. after exposure. The levels of blood ALT and LDH in exposed groups were found to be significantly increased () 24 hr. following exposure. The exposed groups exhibited various degrees of pleural effusion and pericardial effusion. Our findings indicated respiratory exposure to polyacrylate/nanosilica and nanosilica is likely to cause multiple organ toxicity. Ying-Mei Niu, Xiao-Li Zhu, Bing Chang, Zhao-Hui Tong, Wen Cao, Pei-Huan Qiao, Lin-Yuan Zhang, Jing Zhao, and Yu-Guo Song Copyright © 2016 Ying-Mei Niu et al. All rights reserved. Evaluation of Hepatotoxicity with Treatment Doses of Flucytosine and Amphotericin B for Invasive Fungal Infections Tue, 02 Feb 2016 11:28:23 +0000 Invasive fungal infection is a well-known cause of morbidity and mortality in immunocompromised patients. In this study we aimed to evaluate the hepatotoxicity induced by combined therapy of flucytosine and amphotericin B, at three different doses administered to mice for 14 days: 50 mg/kg flucytosine and 300 μg/kg amphotericin B; 100 mg/kg flucytosine and 600 μg/kg amphotericin B; 150 mg/kg flucytosine and 900 μg/kg amphotericin B. Liver injuries were evaluated by analysis of optic and electron microscopy samples, changes in TNF-α, IL-6, and NF-κB inflammation markers levels of expression, and evaluation of mRNA profiles. Histological and ultrastructural analysis revealed an increase in parenchymal and portal inflammation in mice and Kupffer cells activation. Combined antifungal treatment stimulated activation of an inflammatory pathway, demonstrated by a significant dose-dependent increase of TNF-α and IL-6 immunoreactivity, together with mRNA upregulation. Also, NF-κB was activated, as suggested by the high levels found in hepatic tissue and upregulation of target genes. Our results suggest that antifungal combined therapy exerts a synergistic inflammatory activation in a dose-dependent manner, through NF-κB pathway, which promotes an inflammatory cascade during inflammation. The use of combined antifungal therapy needs to be dose limiting due to the associated risk of liver injury, especially for those patients with hepatic dysfunction. Alexandra Folk, Coralia Cotoraci, Cornel Balta, Maria Suciu, Hildegard Herman, Oana Maria Boldura, Sorina Dinescu, Lucian Paiusan, Aurel Ardelean, and Anca Hermenean Copyright © 2016 Alexandra Folk et al. All rights reserved. Early Developmental Low-Dose Methylmercury Exposure Alters Learning and Memory in Periadolescent but Not Young Adult Rats Wed, 13 Jan 2016 12:07:15 +0000 Few studies have assessed the effects of developmental methylmercury (MeHg) exposure on learning and memory at different ages. The possibility of the amelioration or worsening of the effects has not been sufficiently investigated. This study aimed to assess whether low-dose MeHg exposure in utero and during suckling induces differential disturbances in learning and memory of periadolescent and young adult rats. Four experimental groups of pregnant Sprague-Dawley rats were orally exposed to MeHg or vehicle from gestational day 5 to weaning: (1) control (vehicle), (2) 250 μg/kg/day MeHg, (3) 500 μg/kg/day MeHg, and (4) vehicle, and treated on the test day with MK-801 (0.15 mg/kg i.p.), an antagonist of the N-methyl D-aspartate receptor. The effects were evaluated in male offspring through the open field test, object recognition test, Morris water maze, and conditioned taste aversion. For each test and stage assessed, different groups of animals were used. MeHg exposure, in a dose-dependent manner, disrupted exploratory behaviour, recognition memory, spatial learning, and acquisition of aversive memories in periadolescent rats, but alterations were not observed in littermates tested in young adulthood. These results suggest that developmental low-dose exposure to MeHg induces age-dependent detrimental effects. The relevance of decreasing exposure to MeHg in humans remains to be determined. Damaris Albores-Garcia, Leonor C. Acosta-Saavedra, Alberto J. Hernandez, Miriam J. Loera, and Emma S. Calderón-Aranda Copyright © 2016 Damaris Albores-Garcia et al. All rights reserved. Phytochemical Screening and Acute Oral Toxicity Study of Java Tea Leaf Extracts Sun, 27 Dec 2015 10:13:28 +0000 The term Java tea refers to the decoction of Orthosiphon stamineus (OS) Benth (Lamiaceae) leaves, which are widely consumed by the people in Europe and South East Asian countries. The OS leaves are known for their use in traditional medicinal systems as a prophylactic and curative agent for urinary stone, diabetes, and hypertension and also as a diuretic agent. The present study was aimed at evaluating its possible toxicity. Herein, the major phytochemical constituents of microwave dried OS leaf, which is the common drying process for tea sachets in the market, were also identified. The acute oral toxicity test of aqueous, 50% aqueous ethanolic, and ethanolic extracts of OS was performed at a dose of 5000 mg/Kg body weight of Sprague-Dawley rats. During the 14-day study, the animals were observed for any mortality, behavioral, motor-neuronal abnormalities, body weight, and feed-water consumption pattern. The hematological and serum biochemical parameters to assess the kidney and liver functions were carried out, along with the histological analysis of these organs. It was found that all microwave dried OS leaf extracts did not cause any toxic effects or mortality at the administered dose. No abnormality was noticed in all selected parameters in rats of both sexes as compared with their respective control groups. Thus, the possible oral lethal dose for microwave dried Java tea leaves is more than 5000 mg/Kg body weight. Raghunath Pariyani, Intan Safinar Ismail, Amalina Ahmad Azam, Faridah Abas, Khozirah Shaari, and Mohd Roslan Sulaiman Copyright © 2015 Raghunath Pariyani et al. All rights reserved. Zearalenone Inhibits Rat and Human 11β-Hydroxysteroid Dehydrogenase Type 2 Mon, 21 Dec 2015 06:23:18 +0000 Zearalenone is a mycotoxin produced by Fusarium spp. 11β-Hydroxysteroid dehydrogenases, isoforms 1 (HSD11B1) and 2 (HSD11B2), have been demonstrated to be the regulators of the local level of active glucocorticoid, which has a broad range of physiological actions. In the present study, the potency of zearalenone was tested for the inhibition of HSD11B1 and HSD11B2 in rat and human tissues. Zearalenone showed potent inhibition of HSD11B2 with the half-maximal inhibitory concentration (IC50) calculated at 49.63 and 32.22 μM for the rat and human, respectively. Results showed that zearalenone competitively inhibited HSD11B2 when a steroid substrate was used. However, it served as an uncompetitive inhibitory factor when the cofactor NAD+ was used. In contrast, the potency of zearalenone to inhibit both rat and human HSD11B1 was diminished, with the concentration of 100 μM causing almost no inhibitory effect on the isoform. In conclusion, we observed that zearalenone is a selective inhibitor of HSD11B2, implying that this agent may cause excessive glucocorticoid action in local tissues such as kidney and placentas. Linxi Li, Xiaolong Wu, Hongguo Guan, Baiping Mao, Huang Wang, Xiaohuan Yuan, Yanhui Chu, Jianliang Sun, and Ren-Shan Ge Copyright © 2015 Linxi Li et al. All rights reserved. Arsenic and Mercury Containing Traditional Chinese Medicine (Realgar and Cinnabar) Strongly Inhibit Organic Anion Transporters, Oat1 and Oat3, In Vivo in Mice Wed, 16 Dec 2015 07:05:58 +0000 Toxic heavy metals, including mercury (Hg) and arsenic (As), accumulate preferentially in kidneys and always cause acute renal failure. The aim of this study was to investigate whether these samples affect organic anion transporters, Oat1 and Oat3, in vivo in mice kidney. Mice () were orally treated with investigational samples. After last administration, all mice were i.v. p-aminohippuric acid (PAH), and the blood and kidneys samples were collected. The concentrations of PAH were quantified by spectrophotometry. mRNA expressions of Oat1 and Oat3 were assayed by real-time PCR. In comparison with corresponding control, major pharmacokinetic parameters of PAH in sera were significantly changed by investigational samples (), PAH accumulations in the kidney tissues were significantly higher (), PAH uptake by renal slices was greatly reduced, Oat1 and Oat3 mRNA expression were significantly inhibited in investigational sample groups. Arsenic and mercury containing traditional Chinese medicine (Realgar and Cinnabar) probably induce kidney damage through inhibiting several members of the organic anion transporters (such as OAT1 and OAT3). Wen-Hao Yu, Na Zhang, Jin-Feng Qi, Chen Sun, Yong-Hui Wang, and Mei Lin Copyright © 2015 Wen-Hao Yu et al. All rights reserved. Pesticides and Health in Vegetable Production in Kenya Thu, 10 Dec 2015 06:10:29 +0000 This paper investigates the determinants of pesticide-related cost of illness (COI) and acute symptoms, using a balanced panel of 363 farmers interviewed from seven major vegetable producing districts of Kenya. Finding shows that the incidences of pesticide-related health impairments have increased. Variation in number of symptoms and symptom severity significantly explained COI. The personal protective equipment (PPE), education level, record keeping, and geographical location considerably determined health impairments. Encouraging the proper use of PPE and record keeping of pesticide use could greatly reduce poisoning cases and COI. Ibrahim Macharia Copyright © 2015 Ibrahim Macharia. All rights reserved. Overview of Emerging Contaminants and Associated Human Health Effects Wed, 02 Dec 2015 14:20:07 +0000 In recent decades, because of significant progress in the analysis and detection of trace pollutants, emerging contaminants have been discovered and quantified in living beings and diverse environmental substances; however, the adverse effects of environmental exposure on the general population are largely unknown. This review summarizes the conclusions of the comprehensive epidemic literature and representative case reports relevant to emerging contaminants and the human body to address concerns about potential harmful health effects in the general population. The most prevalent emerging contaminants include perfluorinated compounds, water disinfection byproducts, gasoline additives, manufactured nanomaterials, human and veterinary pharmaceuticals, and UV-filters. Rare but statistically meaningful connections have been reported for a number of contaminants and cancer and reproductive risks. Because of contradictions in the outcomes of some investigations and the limited number of articles, no significant conclusions regarding the relationship between adverse effects on humans and extents of exposure can be drawn at this time. Here, we report that the current evidence is not conclusive and comprehensive and suggest prospective cohort studies in the future to evaluate the associations between human health outcomes and emerging environmental contaminants. Meng Lei, Lun Zhang, Jianjun Lei, Liang Zong, Jiahui Li, Zheng Wu, and Zheng Wang Copyright © 2015 Meng Lei et al. All rights reserved. Riboflavin Arrests Cisplatin-Induced Neurotoxicity by Ameliorating Cellular Damage in Dorsal Root Ganglion Cells Wed, 02 Dec 2015 11:43:21 +0000 Cis-Diamminedichloroplatinum II- (CP-) induced neurotoxicity is one of the least explored aspects of this drug. Dorsal root ganglia (DRG) cells are considered as the primary target, and their damage plays a vital role in pathogenesis and etiology of CP-induced neurotoxicity. The present study is aimed at confirming if riboflavin (RF) has any protective role in shielding the DRG from CP-induced toxicity. After conducting the established treatment strategy on mice under photoillumination, it was observed that, despite the fact that RF alone is partially toxic, its combination with CP significantly ameliorated the drug-induced damage in DRG cells as evidenced by histological analysis. In addition, it was interesting to observe that the combination group (RF + CP) was able to induce apoptosis in the target cells up to a significant extent which is considered as the most preferred way of countering cancer cells. Therefore, RF can act as an effective adjuvant compound in CP-based chemoradiotherapy to improve clinical outcomes in the contemporary anticancer treatment regimes. Maria Salman, Imrana Naseem, Iftekhar Hassan, Aijaz A. Khan, and Ibrahim M. Alhazza Copyright © 2015 Maria Salman et al. All rights reserved. PFOS Disturbs BDNF-ERK-CREB Signalling in Association with Increased MicroRNA-22 in SH-SY5Y Cells Sun, 15 Nov 2015 12:33:59 +0000 Perfluorooctane sulfonate (PFOS), a ubiquitous environmental pollutant, is neurotoxic to mammalian species. However, the underlying mechanism of its neurotoxicity was unclear. We hypothesized that PFOS suppresses BDNF expression to produce its neurotoxic effects by inhibiting the ERK-CREB pathway. SH-SY5Y human neuroblastoma cells were exposed to various concentrations of PFOS to examine the role of the BDNF-ERK-CREB signalling pathway in PFOS-induced apoptosis and cytotoxicity. Furthermore, to ascertain the mechanism by which PFOS reduces BDNF signalling, we examined the expression levels of miR-16 and miR-22, which potentially regulate BDNF mRNA translation at the posttranscriptional level. Results indicated that PFOS significantly decreased cell viability and induced apoptosis in SH-SY5Y cells. In addition, BDNF and pERK protein levels decreased after PFOS treatment; however, pCREB protein levels were significantly elevated in PFOS treated groups. TrkB protein expression increased in the 10 μM and 50 μM PFOS groups and significantly decreased in the 100 μM PFOS group. Our results demonstrated that PFOS exposure decreased miR-16 expression and increased miR-22 expression, which may represent a possible mechanism by which PFOS decreases BDNF protein levels. PFOS may inhibit BDNF-ERK-CREB signalling by increasing miR-22 levels, which may, in part, explain the mechanism of PFOS neurotoxicity. Wu Li, Qing-zhi He, Cheng-qiu Wu, Xiao-yuan Pan, Jing Wang, Yan Tan, Xiao-yun Shan, and Huai-cai Zeng Copyright © 2015 Wu Li et al. All rights reserved. Heavy Metal Contents and Physical Parameters of Aegiceras corniculatum, Brassica juncea, and Litchi chinensis Honeys from Bangladesh Wed, 04 Nov 2015 12:09:39 +0000 The present study was undertaken to determine the heavy metal levels and the physicochemical parameters (pH, electrical conductivity (EC), and ash, moisture, and total sugar content) of honeys from Bangladesh. Three different floral honeys were investigated, namely, khalsi (Aegiceras corniculatum), mustard (Brassica juncea), and litchi (Litchi chinensis) honeys. The heavy metals in the honeys were determined by using a High Temperature Dry Oxidation method followed by Atomic Absorption Spectroscopy. The mean pH, EC, and ash, moisture, and total sugar contents of the investigated honeys were 3.6, 0.51 mS/cm, 0.18%, 18.83%, and 68.30%, respectively. Iron was the most abundant among all the investigated heavy metals, ranging from 13.51 to 15.44 mg/kg. The mean concentrations of Mn and Zn in the investigated honeys were 0.28 mg/kg and 2.99 mg/kg, respectively. Cd was below the detection limit, and lead was found in some honey samples, but their contents were below the recommended Maximum Acceptable Level. Cr was also found in all of the samples, but its concentration was within the limit. The physicochemical analysis of the honey samples yielded levels within the limits set by the international honey legislation, indicating that the honey samples were of good quality and had acceptable values for maturity, purity, and freshness. Nandita Sarker, Muhammed Alamgir Zaman Chowdhury, Abu Naieum Muhammad Fakhruddin, Zeenath Fardous, Mohammed Moniruzzaman, and Siew Hua Gan Copyright © 2015 Nandita Sarker et al. All rights reserved. Corrigendum to “Toxicity of Graphene Shells, Graphene Oxide, and Graphene Oxide Paper Evaluated with Escherichia coli Biotests” Thu, 22 Oct 2015 14:06:23 +0000 Ludmila V. Efremova, Alexey S. Vasilchenko, Eduard G. Rakov, and Dmitry G. Deryabin Copyright © 2015 Ludmila V. Efremova et al. All rights reserved. Identification of pLG72-Induced Oxidative Stress Using Systemic Approaches Sun, 11 Oct 2015 09:33:44 +0000 G72 is a schizophrenia-susceptible gene encoding a polypeptide with 153 amino acids. In 2002, it was originally proposed as an activator of D-amino acid oxidase (DAOA) that could enhance the activity of DAAO and subsequently reduce the neurotransmission of N-methyl-D-aspartate receptors. However, several controversial findings have been reported recently. Due to a number of inconsistent descriptions of pLG72’s biofunctions, this study aims to identify the cellular effects induced by pLG72 in U87 cells using systems biology approaches. The analyses of transcriptomics and biological networks showed that pLG72 might be involved in the induction of oxidative stress. To confirm the in silico prediction, we tested and discovered that overexpression of pLG72 effectively enhanced reactive oxygen species (ROS) in U87 cells and, furthermore, this induction can be quenched by Tempol, a general ROS scavenger. Therefore, G72-transgenic mice presenting some psychiatric symptoms, along with the pLG72 level being significantly increased in the serum of patients with schizophrenia, have led us to propose that the ROS enhancement in mental diseases may be from the overexpression of pLG72 in brain cells. Maofeng Wang, Hsi-Ju Chen, Jun Zhang, Weimin Li, Xinyou Xie, and Hao-Teng Chang Copyright © 2015 Maofeng Wang et al. All rights reserved. MicroRNAs-mRNAs Expression Profile and Their Potential Role in Malignant Transformation of Human Bronchial Epithelial Cells Induced by Cadmium Sun, 04 Oct 2015 09:03:29 +0000 Background. Our study was designed to elucidate whether there were miRNA and mRNA aberrantly expression profiles and potential role in malignant transformation of 16HBE induced by Cd. Methods. mRNA and miRNA expression profiles were determined in 35th Cd-induced 16HBE and untreated 16HBE by microarray. A series of bioinformatics analyses such as predicting targets, GO, KEGG were performed to find DEGs, coexpressing networks between miRNAs and mRNAs and its functions. Results. 498 DEGs were found. 8 Cd-responsive novel miRNAs predicted previously were identified, and 5 of them were downregulated. 214 target genes were predicted for the Cd-responsive miRNAs, many of which appeared to regulate gene networks. Target gene CCM2 was showed reciprocal effect by miRNAs. According to the combination analysis, hsa-miR-27b-3p regulated most of the mRNAs, especially upregulated expression genes. The differentially expressed miRNAs are involved in the biological processes and channels, and these GO and KEGG enrichment analyses result were significantly enriched in the Cd-responsive. Discussion. These results provided a tight link for the miRNA-mRNA integrated network and implied the role of novel miRNAs in malignant transformation of 16HBE induced by Cadmium. It is better to understand the novel molecular mechanism of cadmium-induced tumorigenesis. Qun Liu, Chanjiao Zheng, Huanyu Shen, Zhiheng Zhou, and Yixiong Lei Copyright © 2015 Qun Liu et al. All rights reserved. Immunomodulation by Zearalenone in Carp (Cyprinus carpio L.) Tue, 29 Sep 2015 13:50:41 +0000 The mycotoxin zearalenone (ZEN) is a frequent contaminant of animal feeds, but its effects on fish have not yet been investigated extensively. In order to fill this gap a feeding trial with juvenile carp was conducted. Three groups of fish were fed feeds contaminated with ZEN at three concentrations (low: 332 μg kg−1, medium: 621 μg kg−1, and high: 797 μg kg−1 feed) for four weeks. Possible reversible effects of ZEN were evaluated by feeding additional groups with the ZEN-contaminated feeds for four weeks, followed by the uncontaminated diet for two weeks. Immune function of isolated leukocytes from head kidney and trunk kidney was assessed using the assessment of NO production, the respiratory burst assay, the chemiluminescence assay, and the measurement of arginase activities. These investigations frequently revealed increased immune responses after exposure of fish to low ZEN concentrations and reduced immune responses after exposure to high mycotoxin concentrations. Moreover, the feeding of the uncontaminated diet for further two weeks did not improve the immune responses in most cases. These results indicate that cellular immune functions in ZEN-contaminated carp are influenced which may be relevant for fish health in aquaculture. Constanze Pietsch, Ranka Junge, and Patricia Burkhardt-Holm Copyright © 2015 Constanze Pietsch et al. All rights reserved. In Vivo Anti-Candida Activity of Phenolic Extracts and Compounds: Future Perspectives Focusing on Effective Clinical Interventions Mon, 24 Aug 2015 09:48:01 +0000 Candida species have increasingly deserved a special attention among the medical community. In spite of the presence of Candida species as a human commensal, alarming rates of local and systemic infections have been observed, varying from moderate to severe impact. Currently available antifungal drugs have progressively lost their effectiveness, pointing urgently the problem of the microorganisms with acquired-resistance. Natural matrices are secularly used for numerous purposes, being inclusive and highly effective as antimicrobials. Increasing evidence gives a particular emphasis to the contribution of phenolic extracts and related individual compounds. In vitro studies clearly confirm their prominent effects, but the confirmation through in vivo studies, including the involved mechanisms of action, is not so much deepened. Therefore, the present report aims to provide extensive knowledge about all these aspects, highlighting the most efficient phytochemical formulations, including therapeutic doses. Further studies need to be incited to deepen knowledge on this area, namely, focused on clinical trials to provide safer and more effective antimicrobials than the current ones. Natália Martins, Lillian Barros, Mariana Henriques, Sónia Silva, and Isabel C. F. R. Ferreira Copyright © 2015 Natália Martins et al. All rights reserved. The Effect of MGCD0103 on CYP450 Isoforms Activity of Rats by Cocktail Method Wed, 19 Aug 2015 09:13:22 +0000 MGCD0103, an isotype-selective histone deacetylase inhibitor (HDACi), has been clinically evaluated for the treatment of hematologic malignancies and advanced solid tumors, alone and in combination with standard-of-care agents. In order to investigate the effects of MGCD0103 on the metabolic capacity of cytochrome P450 (CYP) enzymes, a cocktail method was employed to evaluate the activities of human CYP2B1, CYP1A2, CYP2C11, CYP2D6, CYP3A4, and CYP2C9. The rats were randomly divided into MGCD0103 group (Low, Medium, and High) and control group. The MGCD0103 group rats were given 20, 40, and 80 mg/kg (Low, Medium, and High) MGCD0103 by continuous intragastric administration for 7 days. Six probe drugs, bupropion, phenacetin, tolbutamide, metoprolol, testosterone, and omeprazole, were given to rats through intragastric administration, and the plasma concentrations were determined by UPLC-MS/MS. Statistical pharmacokinetics difference for tolbutamide in rats were observed by comparing MGCD0103 group with control group. Continuous 7-day intragastric administration of MGCD0103 slightly induces the activities of CYP2C11 of rats. Jinzhang Cai, Qingwei Zhang, Kezhi Lin, Lufeng Hu, and Yuancai Zheng Copyright © 2015 Jinzhang Cai et al. All rights reserved. Degradation Effect of Sulfa Antibiotics by Potassium Ferrate Combined with Ultrasound (Fe(VI)-US) Tue, 11 Aug 2015 14:18:41 +0000 Sulfa antibiotics are a family of typical broad-spectrum antibiotics, which have become one of the most frequently detected antibiotics in water, posing a great threat to human health and ecosystem. Potassium ferrate is a new type of high-efficiency multifunctional water treatment agent, collecting the effects of oxidation, adsorption, flocculation, coagulation, sterilization, and deodorization. Performance and mechanism of degradation of typical broad-spectrum antibiotics by Fe(VI)-US were further studied, investigating the degradation effect of sulfa antibiotics by single ultrasound, single potassium ferrate, and potassium ferrate-ultrasound (Fe(VI)-US). It was found that Fe(VI)-US technology had a significant role in promoting the degradation of sulfa antibiotics via orthogonal experiments. Factors evaluated included sulfa antibiotics type, pH value, potassium ferrate dosage, ultrasonic frequency, and ultrasonic power, with the pH value and potassium ferrate dosage being affected most significantly. One reason for synergy facilitating the degradation is the common oxidation of potassium ferrate and ultrasound, and the other is that Fe(III) produced promotes the degradation rate. According to the product analysis and degradation pathways of three sulfa antibiotics, ferrate-sonication sulfa antibiotics are removed by hydroxyl radical oxidation. Kejia Zhang, Zhang Luo, Tuqiao Zhang, Naiyun Gao, and Yan Ma Copyright © 2015 Kejia Zhang et al. All rights reserved. Oxidation of Sulfonamides in Aqueous Solution by UV-TiO2-Fe(VI) Tue, 11 Aug 2015 12:50:57 +0000 The photocatalytic degradation of sulfonamides in aqueous TiO2 suspension under UV irradiation has been investigated using potassium ferrate as electron acceptors. The results showed that the stability of Fe(VI) is dependent on pH significantly, and the stability reduces obviously in the presence of UV-TiO2. The experiments indicated that Fe(VI) could effectively scavenge the conduction band electrons from the surface of TiO2. The photocatalytic oxidation of sulfonamides with Fe(VI) was found to be much faster than that without Fe(VI). The SD, SM, and SMX concentration was greatly reduced by 89.2%, 83.4%, and 82.0%, respectively, after 10 min with UV-TiO2-Fe(VI), comparing to 65.2%, 66.0%, and 71.9%, respectively, with Fe(VI) only in the dark and 71.3%, 72.7%, and 76.0%, respectively, with UV-TiO2. The pH value of solution significantly influenced the sulfonamides degradation in UV-TiO2-Fe(VI) system. The degradation amount of sulfonamides after 10 min was a maximum at pH 7. The intermediate products of sulfonamides oxidation by UV-TiO2-Fe(VI) were analysed by LC-HESI-MS-MS and the results suggested that a majority of sulfonamides turned into large-molecule products without complete mineralization. Yan Ma, Kejia Zhang, Cong Li, Tuqiao Zhang, and Naiyun Gao Copyright © 2015 Yan Ma et al. All rights reserved. Advanced Treatment of Pesticide-Containing Wastewater Using Fenton Reagent Enhanced by Microwave Electrodeless Ultraviolet Tue, 11 Aug 2015 12:34:51 +0000 The photo-Fenton reaction is a promising method to treat organic contaminants in water. In this paper, a Fenton reagent enhanced by microwave electrodeless ultraviolet (MWEUV/Fenton) method was proposed for advanced treatment of nonbiodegradable organic substance in pesticide-containing biotreated wastewater. MWEUV lamp was found to be more effective for chemical oxygen demand (COD) removal than commercial mercury lamps in the Fenton process. The pseudo-first order kinetic model can well describe COD removal from pesticide-containing wastewater by MWEUV/Fenton, and the apparent rate constant (k) was 0.0125 min−1. The optimal conditions for MWEUV/Fenton process were determined as initial pH of 5, Fe2+ dosage of 0.8 mmol/L, and H2O2 dosage of 100 mmol/L. Under the optimal conditions, the reaction exhibited high mineralization degrees of organics, where COD and dissolved organic carbon (DOC) concentration decreased from 183.2 mg/L to 36.9 mg/L and 43.5 mg/L to 27.8 mg/L, respectively. Three main pesticides in the wastewater, as Dimethoate, Triazophos, and Malathion, were completely removed by the MWEUV/Fenton process within 120 min. The high degree of pesticides decomposition and mineralization was proved by the detected inorganic anions. Gong Cheng, Jing Lin, Jian Lu, Xi Zhao, Zhengqing Cai, and Jie Fu Copyright © 2015 Gong Cheng et al. All rights reserved. Real versus Simulated Mobile Phone Exposures in Experimental Studies Tue, 04 Aug 2015 13:06:34 +0000 We examined whether exposures to mobile phone radiation in biological/clinical experiments should be performed with real-life Electromagnetic Fields (EMFs) emitted by commercially available mobile phone handsets, instead of simulated EMFs emitted by generators or test phones. Real mobile phone emissions are constantly and unpredictably varying and thus are very different from simulated emissions which employ fixed parameters and no variability. This variability is an important parameter that makes real emissions more bioactive. Living organisms seem to have decreased defense against environmental stressors of high variability. While experimental studies employing simulated EMF-emissions present a strong inconsistency among their results with less than 50% of them reporting effects, studies employing real mobile phone exposures demonstrate an almost 100% consistency in showing adverse effects. This consistency is in agreement with studies showing association with brain tumors, symptoms of unwellness, and declines in animal populations. Average dosimetry in studies with real emissions can be reliable with increased number of field measurements, and variation in experimental outcomes due to exposure variability becomes less significant with increased number of experimental replications. We conclude that, in order for experimental findings to reflect reality, it is crucially important that exposures be performed by commercially available mobile phone handsets. Dimitris J. Panagopoulos, Olle Johansson, and George L. Carlo Copyright © 2015 Dimitris J. Panagopoulos et al. All rights reserved. Genetic and Epigenetic Effects of Environmental Mutagens and Carcinogens Tue, 04 Aug 2015 12:03:28 +0000 Alessandra Pulliero, Jia Cao, Luciana dos Reis Vasques, and Francesca Pacchierotti Copyright © 2015 Alessandra Pulliero et al. All rights reserved. Effect of Environmental Chemical Stress on Nuclear Noncoding RNA Involved in Epigenetic Control Mon, 03 Aug 2015 12:11:08 +0000 In the last decade the role of noncoding RNAs (ncRNAs) emerges not only as key elements of posttranscriptional gene silencing, but also as important players of epigenetic regulation. New kind and new functions of ncRNAs are continuously discovered and one of their most important roles is the mediation of environmental signals, both physical and chemical. The activity of cytoplasmic short ncRNA is extensively studied, in spite of the fact that their function and role in the nuclear compartment are not yet completely unraveled. Cellular nucleus contains a multiplicity of long and short ncRNAs controlling at different levels transcriptional and epigenetic processes. In addition, some ncRNAs are involved in RNA editing and quality control. In this paper we review the existing knowledge dealing with how chemical stressors can influence the functionality of short nuclear ncRNAs. Furthermore, we perform bioinformatics analyses indicating that chemical environmental stressors not only induce DNA damage but also influence the mechanism of ncRNAs production and control. Patrizio Arrigo and Alessandra Pulliero Copyright © 2015 Patrizio Arrigo and Alessandra Pulliero. All rights reserved. Association between Genetic Polymorphisms of DNA Repair Genes and Chromosomal Damage for 1,3-Butadiene-Exposed Workers in a Matched Study in China Mon, 03 Aug 2015 12:09:58 +0000 The aim of the study was to examine the association between polymorphisms of DNA repair genes and chromosomal damage of 1,3-butadiene- (BD-) exposed workers. The study was conducted in 45 pairs of occupationally exposed workers in a BD product workshop and matched control workers in an administrative office and a circulatory water workshop in China. Newly developed biomarkers (micronuclei, MNi; nucleoplasmic bridges, NPBs; nuclear buds, NBUDs) in the cytokinesis-blocked micronucleus (CBMN) cytome assay were adopted to detect chromosomal damage. PCR and PCR-restriction fragment length polymorphism (RFLP) are adopted to analyze polymorphisms of DNA repair genes, such as X-ray repair cross-complementing Group 1 (XRCC1), O6-methylguanine-DNA methyltransferase (MGMT), poly (adenosine diphosphate-ribose) polymerases (ADPRT), and apurinic/apyrimidinic endonucleases (APE1). The BD-exposed workers exhibited increased frequencies of MNi and NPBs when compared to subjects in the control group. The results also show that the BD-exposed workers carrying XRCC1 diplotypes TCGA-CCGG () (, 95% CI: 1.03–4.28) and TCGG-TCGA () (, 95% CI: 0.76–2.65) had statistically higher NBUD frequencies than those who carried diplotype TCGG-TCGG (). Our study suggests that polymorphisms of XRCC1 gene may influence chromosomal damage in BD-exposed workers. Menglong Xiang, Lei Sun, Xiaomei Dong, Huan Yang, Wen-bin Liu, Niya Zhou, Xue Han, Ziyuan Zhou, Zhihong Cui, Jing-yi Liu, Jia Cao, and Lin Ao Copyright © 2015 Menglong Xiang et al. All rights reserved. Applying a Weight-of-Evidence Approach to Evaluate Relevance of Molecular Landscapes in the Exposure-Disease Paradigm Mon, 03 Aug 2015 11:34:50 +0000 Information on polymorphisms, mutations, and epigenetic events has become increasingly important in our understanding of molecular mechanisms associated with exposures-disease outcomes. Molecular landscapes can be developed to illustrate the molecular characteristics for environmental carcinogens as well as associated disease outcomes, although comparison of these molecular landscapes can often be difficult to navigate. We developed a method to organize these molecular data that uses a weight-of-evidence approach to rank overlapping molecular events by relative importance for susceptibility to an exposure-disease paradigm. To illustrate the usefulness of this approach, we discuss the example of benzene as an environmental carcinogen and myelodysplastic syndrome (MDS) as a causative disease endpoint. Using this weight-of-evidence method, we found overlapping polymorphisms in the genes for the metabolic enzymes GST and NQO1, both of which may infer risk of benzene-induced MDS. Polymorphisms in the tumor suppressor gene, TP53, and the inflammatory cytokine gene, TNF-α, were also noted, albeit inferring opposing outcomes. The alleles identified in the DNA repair gene RAD51 indicated an increased risk for MDS in MDS patients and low blood cell counts in benzene-exposed workers. We propose the weight-of-evidence approach as a tool to assist in organizing the sea of emerging molecular data in exposure-disease paradigms. Sherilyn A. Gross and Kristen M. Fedak Copyright © 2015 Sherilyn A. Gross and Kristen M. Fedak. All rights reserved. Heterogeneity of Genetic Damage in Cervical Nuclei and Lymphocytes in Women with Different Levels of Dysplasia and Cancer-Associated Risk Factors Mon, 03 Aug 2015 11:15:50 +0000 The comet assay can be used to assess genetic damage, but heterogeneity in the length of the tails is frequently observed. The aims of this study were to evaluate genetic damage and heterogeneity in the cervical nuclei and lymphocytes from patients with different levels of dysplasia and to determine the risk factors associated with the development of cervical cancer. The study included 97 females who presented with different levels of dysplasia. A comet assay was performed in peripheral blood lymphocytes and cervical epithelial cells. Significant genetic damage () was observed only in patients diagnosed with nuclei cervical from dysplasia III (NCDIII) and lymphocytes from dysplasia I (LDI). However, the standard deviations of the tail lengths in the cervical nuclei and lymphocytes from patients with dysplasia I were significantly different () from the standard deviations of the tail lengths in the nuclei cervical and lymphocytes from patients with DII and DIII (NCDII, NCDIII and LDII, LDIII), indicating a high heterogeneity in tail length. Results suggest that genetic damage could be widely present but only manifested as increased tail length in certain cell populations. This heterogeneity could obscure the statistical significance of the genetic damage. Carlos Alvarez-Moya, Mónica Reynoso-Silva, Alejandro A. Canales-Aguirre, José O. Chavez-Chavez, Hugo Castañeda-Vázquez, and Alfredo I. Feria-Velasco Copyright © 2015 Carlos Alvarez-Moya et al. All rights reserved. Oxidative Stress Alters miRNA and Gene Expression Profiles in Villous First Trimester Trophoblasts Mon, 03 Aug 2015 10:45:10 +0000 The relationship between oxidative stress and miRNA changes in placenta as a potential mechanism involved in preeclampsia (PE) is not fully elucidated. We investigated the impact of oxidative stress on miRNAs and mRNA expression profiles of genes associated with PE in villous 3A first trimester trophoblast cells exposed to H2O2 at 12 different concentrations (0-1 mM) for 0.5, 4, 24, and 48 h. Cytotoxicity, determined using the SRB assay, was used to calculate the IC50 of H2O2. RNA was extracted after 4 h exposure to H2O2 for miRNA and gene expression profiling. H2O2 exerted a concentration- and time-dependent cytotoxicity on 3A trophoblast cells. Short-term exposure of 3A cells to low concentration of H2O2 (5% of IC50) significantly altered miRNA profile as evidenced by significant changes in 195 out of 595 evaluable miRNAs. Tool for annotations of microRNAs (TAM) analysis indicated that these altered miRNAs fall into 43 clusters and 34 families, with 41 functions identified. Exposure to H2O2 altered mRNA expression of 22 out of 84 key genes involved in dysregulation of placental development. In conclusion, short-term exposure of villous first trimester trophoblasts to low concentrations of H2O2 significantly alters miRNA profile and expression of genes implicated in placental development. Courtney E. Cross, Mai F. Tolba, Catherine M. Rondelli, Meixiang Xu, and Sherif Z. Abdel-Rahman Copyright © 2015 Courtney E. Cross et al. All rights reserved. Bisphenol A Effects on Mammalian Oogenesis and Epigenetic Integrity of Oocytes: A Case Study Exploring Risks of Endocrine Disrupting Chemicals Mon, 03 Aug 2015 09:04:46 +0000 Bisphenol A (BPA), originally developed as a synthetic oestrogen, is nowadays extensively used in the production of polymeric plastics. Under harsh conditions, these plastics may release BPA, which then can leach into the environment. Detectable concentrations of BPA have been measured in most analysed samples of human serum, plasma, or urine, as well as in follicular fluid, foetal serum, and amniotic fluid. Here we summarize the evidence about adverse BPA effects on the genetic and epigenetic integrity of mammalian oocytes. We conclude that increasing evidence supports the notion that low BPA concentrations adversely affect the epigenome of mammalian female germ cells, with functional consequences on gene expression, chromosome dynamics in meiosis, and oocyte development. Specific time windows, during which profound chromatin remodelling occurs and maternal imprints are established or protected, appear particularly vulnerable to epigenetic deregulation by BPA. Transgenerational effects have been also observed in the offspring of BPA-treated rodents, although the epigenetic mechanisms of inheritance still need to be clarified. The relevance of these findings for human health protection still needs to be fully assessed, but they warrant further investigation in both experimental models and humans. Ursula Eichenlaub-Ritter and Francesca Pacchierotti Copyright © 2015 Ursula Eichenlaub-Ritter and Francesca Pacchierotti. All rights reserved. Environmental Impact on DNA Methylation in the Germline: State of the Art and Gaps of Knowledge Mon, 03 Aug 2015 08:17:32 +0000 The epigenome consists of chemical changes in DNA and chromatin that without modifying the DNA sequence modulate gene expression and cellular phenotype. The epigenome is highly plastic and reacts to changing external conditions with modifications that can be inherited to daughter cells and across generations. Whereas this innate plasticity allows for adaptation to a changing environment, it also implies the potential of epigenetic derailment leading to so-called epimutations. DNA methylation is the most studied epigenetic mark. DNA methylation changes have been associated with cancer, infertility, cardiovascular, respiratory, metabolic, immunologic, and neurodegenerative pathologies. Experiments in rodents demonstrate that exposure to a variety of chemical stressors, occurring during the prenatal or the adult life, may induce DNA methylation changes in germ cells, which may be transmitted across generations with phenotypic consequences. An increasing number of human biomonitoring studies show environmentally related DNA methylation changes mainly in blood leukocytes, whereas very few data have been so far collected on possible epigenetic changes induced in the germline, even by the analysis of easily accessible sperm. In this paper, we review the state of the art on factors impinging on DNA methylation in the germline, highlight gaps of knowledge, and propose priorities for future studies. Francesca Pacchierotti and Marcello Spanò Copyright © 2015 Francesca Pacchierotti and Marcello Spanò. All rights reserved. Putative Epimutagens in Maternal Peripheral and Cord Blood Samples Identified Using Human Induced Pluripotent Stem Cells Mon, 03 Aug 2015 07:34:50 +0000 The regulation of transcription and genome stability by epigenetic systems are crucial for the proper development of mammalian embryos. Chemicals that disturb epigenetic systems are termed epimutagens. We previously performed chemical screening that focused on heterochromatin formation and DNA methylation status in mouse embryonic stem cells and identified five epimutagens: diethyl phosphate (DEP), mercury (Hg), cotinine, selenium (Se), and octachlorodipropyl ether (S-421). Here, we used human induced pluripotent stem cells (hiPSCs) to confirm the effects of 20 chemicals, including the five epimutagens, detected at low concentrations in maternal peripheral and cord blood samples. Of note, these individual chemicals did not exhibit epimutagenic activity in hiPSCs. However, because the fetal environment contains various chemicals, we evaluated the effects of combined exposure to chemicals (DEP, Hg, cotinine, Se, and S-421) on hiPSCs. The combined exposure caused a decrease in the number of heterochromatin signals and aberrant DNA methylation status at multiple gene loci in hiPSCs. The combined exposure also affected embryoid body formation and neural differentiation from hiPSCs. Therefore, DEP, Hg, cotinine, Se, and S-421 were defined as an “epimutagen combination” that is effective at low concentrations as detected in maternal peripheral and cord blood. Yoshikazu Arai, Koji Hayakawa, Daisuke Arai, Rie Ito, Yusuke Iwasaki, Koichi Saito, Kazuhiko Akutsu, Satoshi Takatori, Rie Ishii, Rumiko Hayashi, Shun-Ichiro Izumi, Norihiro Sugino, Fumio Kondo, Masakazu Horie, Hiroyuki Nakazawa, Tsunehisa Makino, Mitsuko Hirosawa, Kunio Shiota, and Jun Ohgane Copyright © 2015 Yoshikazu Arai et al. All rights reserved. Noncoding RNAs: Possible Players in the Development of Fluorosis Mon, 03 Aug 2015 07:22:33 +0000 Fluorosis is caused by excess of fluoride intake over a long period of time. Aberrant change in the Runt-related transcription factor 2 (RUNX2) mediated signaling cascade is one of the decisive steps during the pathogenesis of fluorosis. Up to date, role of fluoride on the epigenetic alterations is not studied. In the present study, global expression profiling of short noncoding RNAs, in particular miRNAs and snoRNAs, was carried out in sodium fluoride (NaF) treated human osteosarcoma (HOS) cells to understand their possible role in the development of fluorosis. qPCR and in silico hybridization revealed that miR-124 and miR-155 can be directly involved in the transcriptional regulation of Runt-related transcription factor 2 (RUNX2) and receptor activator of nuclear factor κ-B ligand (RANKL) genes. Compared to control, C/D box analysis revealed marked elevation in the number of UG dinucleotides and D-box sequences in NaF exposed HOS cells. Herein, we report miR-124 and miR-155 as the new possible players involved in the development of fluorosis. We show that the alterations in UG dinucleotides and D-box sequences of snoRNAs could be due to NaF exposure. Atul P. Daiwile, Saravanadevi Sivanesan, Alberto Izzotti, Amit Bafana, Pravin K. Naoghare, Patrizio Arrigo, Hemant J. Purohit, Devendra Parmar, and Krishnamurthi Kannan Copyright © 2015 Atul P. Daiwile et al. All rights reserved. Genotoxic Effect in Autoimmune Diseases Evaluated by the Micronucleus Test Assay: Our Experience and Literature Review Mon, 03 Aug 2015 06:59:11 +0000 Autoimmune diseases (AD) are classified into organ-specific, systemic, and mixed; all forms of AD share a high risk for cancer development. In AD a destructive immune response induced by autoreactive lymphocytes is started and continues with the production of autoantibodies against different targets; furthermore apoptosis failure and loss of balance in oxidative stress as a consequence of local or systemic inflammation are common features seen in AD as well. Micronucleus (MN) assay can be performed in order to evaluate loss of genetic material in a clear, accurate, fast, simple, and minimally invasive test. The MN formation in the cytoplasm of cells that have undergone proliferation is a consequence of DNA fragmentation during mitosis and the appearance of small additional nuclei during interphase. The MN test, widely accepted for in vitro and in vivo genotoxicity research, provides a sensitive marker of genomic damage associated to diverse conditions. In here, we present a review of our work and other published papers concerning genotoxic effect in AD, identified by means of the MN assay, with the aim of proposing this tool as a possible early biomarker for genotoxic damage, which is a consequence of disease progression. Additionally this biomarker could be used for follow-up, to asses genome damage associated to therapies. Olivia Torres-Bugarín, Nicole Macriz Romero, María Luisa Ramos Ibarra, Aurelio Flores-García, Penélope Valdez Aburto, and María Guadalupe Zavala-Cerna Copyright © 2015 Olivia Torres-Bugarín et al. All rights reserved. Effect of 50 Hz Extremely Low-Frequency Electromagnetic Fields on the DNA Methylation and DNA Methyltransferases in Mouse Spermatocyte-Derived Cell Line GC-2 Mon, 03 Aug 2015 06:53:08 +0000 Previous studies have shown that the male reproductive system is one of the most sensitive organs to electromagnetic radiation. However, the biological effects and molecular mechanism are largely unclear. Our study was designed to elucidate the epigenetic effects of 50 Hz ELF-EMF in vitro. Mouse spermatocyte-derived GC-2 cell line was exposed to 50 Hz ELF-EMF (5 min on and 10 min off) at magnetic field intensity of 1 mT, 2 mT, and 3 mT with an intermittent exposure for 72 h. We found that 50 Hz ELF-EMF exposure decreased genome-wide methylation at 1 mT, but global methylation was higher at 3 mT compared with the controls. The expression of DNMT1 and DNMT3b was decreased at 1 mT, and 50 Hz ELF-EMF can increase the expression of DNMT1 and DNMT3b of GC-2 cells at 3 mT. However, 50 Hz ELF-EMF had little influence on the expression of DNMT3a. Then, we established DNA methylation and gene expression profiling and validated some genes with aberrant DNA methylation and expression at different intensity of 50 Hz ELF-EMF. These results suggest that the alterations of genome-wide methylation and DNMTs expression may play an important role in the biological effects of 50 Hz ELF-EMF exposure. Yong Liu, Wen-bin Liu, Kai-jun Liu, Lin Ao, Julia Li Zhong, Jia Cao, and Jin-yi Liu Copyright © 2015 Yong Liu et al. All rights reserved. Environmental Epigenetics: Crossroad between Public Health, Lifestyle, and Cancer Prevention Mon, 03 Aug 2015 06:52:48 +0000 Epigenetics provides the key to transform the genetic information into phenotype and because of its reversibility it is considered an ideal target for therapeutic interventions. This paper reviews the basic mechanisms of epigenetic control: DNA methylation, histone modifications, chromatin remodeling, and ncRNA expression and their role in disease development. We describe also the influence of the environment, lifestyle, nutritional habits, and the psychological influence on epigenetic marks and how these factors are related to cancer and other diseases development. Finally we discuss the potential use of natural epigenetic modifiers in the chemoprevention of cancer to link together public health, environment, and lifestyle. Massimo Romani, Maria Pia Pistillo, and Barbara Banelli Copyright © 2015 Massimo Romani et al. All rights reserved. Impact of Cadmium on Intracellular Zinc Levels in HepG2 Cells: Quantitative Evaluations and Molecular Effects Mon, 03 Aug 2015 06:45:05 +0000 Cadmium is classified as a human carcinogen, and its disturbance in zinc homeostasis has been well established. However, its extent as well as molecular mechanisms involved in cadmium carcinogenesis has yet to be fully clarified. To this end, we used the zinc specific probe Zinquin to visualize and to quantitatively evaluate changes in the concentration of labile zinc, in an in vitro model of human hepatic cells (HepG2) exposed to cadmium. A very large increase (+93%) of intracellular labile zinc, displaced by cadmium from the zinc proteome, was measured when HepG2 were exposed to 10 µM cadmium for 24 hrs. Microarray expression profiling showed that in cells, featuring an increase of labile zinc after cadmium exposure, one of the top regulated genes is Snail1 (+3.6), which is included in the adherens junction pathway and linked to cancer. In the same pathway MET, TGF-βR, and two members of the Rho-family GTPase, Rac, and cdc42 all implicated in the loss of adherence features and acquisition of migratory and cancer properties were regulated, as well. The microRNAs analysis showed a downregulation of miR-34a and miR-200a, both implicated in the epithelial-mesenchymal transition. These microRNAs results support the role played by zinc in affecting gene expression at the posttranscriptional level. Chiara Urani, Pasquale Melchioretto, Maurizio Bruschi, Marco Fabbri, Maria Grazia Sacco, and Laura Gribaldo Copyright © 2015 Chiara Urani et al. All rights reserved. Pulmonary Responses of Sprague-Dawley Rats in Single Inhalation Exposure to Graphene Oxide Nanomaterials Thu, 30 Jul 2015 12:01:33 +0000 Graphene is receiving increased attention due to its potential widespread applications in future. However, the health effects of graphene have not yet been well studied. Therefore, this study examined the pulmonary effects of graphene oxide using male Sprague-Dawley rats and a single 6-hour nose-only inhalation technique. Following the exposure, the rats were allowed to recover for 1 day, 7 days, or 14 days. A total of three groups were compared: control (fresh air), low concentration ( mg/m3), and high concentration ( mg/m3). The exposure to graphene oxide did not induce significant changes in the body weights, organ weights, and food consumption during the 14 days of recovery time. The microalbumin and lactate dehydrogenase levels in the bronchoalveolar lavage (BAL) fluid were not significantly changed due to the exposure. Similarly, total cell count, macrophages, polymorphonuclear leukocytes, and lymphocytes were not significantly altered in the BAL fluid. Plus, the histopathological examination of the rat lungs only showed an uptake of graphene oxide in the alveolar macrophages of the high-concentration group. Therefore, these results demonstrate that the single inhalation exposure to graphene oxide induce minimal toxic responses in rat lungs at the concentrations and time points used in the present study. Sung Gu Han, Jin Kwon Kim, Jae Hoon Shin, Joo Hwan Hwang, Jong Seong Lee, Tae-Gyu Kim, Ji Hyun Lee, Gun Ho Lee, Keun Soo Kim, Heon Sang Lee, Nam Woong Song, Kangho Ahn, and Il Je Yu Copyright © 2015 Sung Gu Han et al. All rights reserved. Cytotoxic and Antitumor Activity of Sulforaphane: The Role of Reactive Oxygen Species Mon, 22 Jun 2015 09:57:06 +0000 According to recent estimates, cancer continues to remain the second leading cause of death and is becoming the leading one in old age. Failure and high systemic toxicity of conventional cancer therapies have accelerated the identification and development of innovative preventive as well as therapeutic strategies to contrast cancer-associated morbidity and mortality. In recent years, increasing body of in vitro and in vivo studies has underscored the cancer preventive and therapeutic efficacy of the isothiocyanate sulforaphane. In this review article, we highlight that sulforaphane cytotoxicity derives from complex, concurring, and multiple mechanisms, among which the generation of reactive oxygen species has been identified as playing a central role in promoting apoptosis and autophagy of target cells. We also discuss the site and the mechanism of reactive oxygen species’ formation by sulforaphane, the toxicological relevance of sulforaphane-formed reactive oxygen species, and the death pathways triggered by sulforaphane-derived reactive oxygen species. Piero Sestili and Carmela Fimognari Copyright © 2015 Piero Sestili and Carmela Fimognari. All rights reserved. Occupational and Environmental Health Effects of Nanomaterials Wed, 10 Jun 2015 09:14:33 +0000 Il Je Yu, Mary Gulumian, Sehyun Shin, Tae Hyun Yoon, and Vladimir Murashov Copyright © 2015 Il Je Yu et al. All rights reserved. Multiwall Carbon Nanotube-Induced Apoptosis and Antioxidant Gene Expression in the Gills, Liver, and Intestine of Oryzias latipes Wed, 03 Jun 2015 12:36:44 +0000 Multiwall carbon nanotubes (MWCNTs) have many attractive properties with potential applications in various fields. Despite their usefulness, however, the associated waste can be hazardous to the environment. To examine adverse effects in aquatic environments, Oryzias latipes were exposed to MWCNTs dispersed in water for 14 days and apoptosis and antioxidant gene expression were observed. This work showed that in gills exposed to 100 mg/L MWCNTs for 4 days, there was significant p53, caspase-3 (Cas3), caspase-8 (Cas8), and caspase-9 (Cas9) gene expression relative to the controls, while catalase (CAT) and glutathione-S-transferase (GST) expression were reduced. At 14 days, CAT, GST, and metallothionein (MT) were induced significantly in the gills and Cas3, Cas8, and Cas9 were induced in the liver. No significant gene induction was seen in intestine. Intracellular reactive oxygen species (ROS) were increased significantly only at 14 days. Histologically, no apoptosis was observed with exposure to 100 mg/L MWCNTs for 21 days. The gills were more sensitive to MWCNT toxicity than the other organs. Males had higher apoptosis gene induction than females. These results demonstrated that MWCNTs could cause apoptosis in a manner influenced by tissue and gender in aqueous environments. Jin Wuk Lee, Young Chul Choi, Rosa Kim, and Sung Kyu Lee Copyright © 2015 Jin Wuk Lee et al. All rights reserved. Quantification of Internalized Silica Nanoparticles via STED Microscopy Mon, 01 Jun 2015 13:55:42 +0000 The development of safe engineered nanoparticles (NPs) requires a detailed understanding of their interaction mechanisms on a cellular level. Therefore, quantification of NP internalization is crucial to predict the potential impact of intracellular NP doses, providing essential information for risk assessment as well as for drug delivery applications. In this study, the internalization of 25 nm and 85 nm silica nanoparticles (SNPs) in alveolar type II cells (A549) was quantified by application of super-resolution STED (stimulated emission depletion) microscopy. Cells were exposed to equal particle number concentrations ( particles mL−1) of each particle size and the sedimentation of particles during exposure was taken into account. Microscopy images revealed that particles of both sizes entered the cells after 5 h incubation in serum supplemented and serum-free medium. According to the in vitro sedimentation, diffusion, and dosimetry (ISDD) model 20–27% of the particles sedimented. In comparison, 102-103 NPs per cell were detected intracellularly serum-containing medium. Furthermore, in the presence of serum, no cytotoxicity was induced by the SNPs. In serum-free medium, large agglomerates of both particle sizes covered the cells whereas only high concentrations (≥ 3.8 × 1012 particles mL−1) of the smaller particles induced cytotoxicity. Henrike Peuschel, Thomas Ruckelshausen, Christian Cavelius, and Annette Kraegeloh Copyright © 2015 Henrike Peuschel et al. All rights reserved. Micro- and Nanosized Particles in Nasal Mucosa: A Pilot Study Mon, 01 Jun 2015 12:23:05 +0000 Objective. The aim of this prospective study is to evaluate presence and quantity of micro- and nanosized particles (NPs) and interindividual differences in their distribution and composition in nasal mucosa. Methods. Six samples of nasal mucosa obtained by mucotomy from patients with chronic hypertrophic rhinosinusitis were examined. Samples divided into 4 parts according to the distance from the nostrils were analyzed by scanning electron microscopy and Raman microspectroscopy to detect solid particles and characterize their morphology and composition. A novel method of quantification of the particles was designed and used to evaluate interindividual differences in distribution of the particles. The findings were compared with patients’ employment history. Results. In all the samples, NPs of different elemental composition were found (iron, barium, copper, titanium, etc.), predominantly in the parts most distant from nostrils, in various depths from the surface of the mucosa and interindividual differences in their quantity and composition were found, possibly in relation to professional exposition. Conclusions. This study has proven the possibility of quantification of distribution of micro- and nanosized particles in tissue samples and that the NPs may deposit in deeper layers of mucosa and their elemental composition may be related to professional exposition to the sources of NPs. Lenka Čábalová, Kristina Čabanová, Hana Bielniková, Jana Kukutschová, Jana Dvořáčková, Kateřina Dědková, Karol Zeleník, and Pavel Komínek Copyright © 2015 Lenka Čábalová et al. All rights reserved. Workplace Exposure to Titanium Dioxide Nanopowder Released from a Bag Filter System Mon, 01 Jun 2015 10:01:26 +0000 Many researchers who use laboratory-scale synthesis systems to manufacture nanomaterials could be easily exposed to airborne nanomaterials during the research and development stage. This study used various real-time aerosol detectors to investigate the presence of nanoaerosols in a laboratory used to manufacture titanium dioxide (TiO2). The TiO2 nanopowders were produced via flame synthesis and collected by a bag filter system for subsequent harvesting. Highly concentrated nanopowders were released from the outlet of the bag filter system into the laboratory. The fractional particle collection efficiency of the bag filter system was only 20% at particle diameter of 100 nm, which is much lower than the performance of a high-efficiency particulate air (HEPA) filter. Furthermore, the laboratory hood system was inadequate to fully exhaust the air discharged from the bag filter system. Unbalanced air flow rates between bag filter and laboratory hood systems could result in high exposure to nanopowder in laboratory settings. Finally, we simulated behavior of nanopowders released in the laboratory using computational fluid dynamics (CFD). Jun Ho Ji, Jong Bum Kim, Gwangjae Lee, Jung-Hun Noh, Se-Jin Yook, So-Hye Cho, and Gwi-Nam Bae Copyright © 2015 Jun Ho Ji et al. All rights reserved. Metal-Based Nanoparticles and the Immune System: Activation, Inflammation, and Potential Applications Mon, 01 Jun 2015 09:17:43 +0000 Nanomaterials, including metal-based nanoparticles, are used for various biological and medical applications. However, metals affect immune functions in many animal species including humans. Different physical and chemical properties induce different cellular responses, such as cellular uptake and intracellular biodistribution, leading to the different immune responses. The goals of this review are to summarize and discuss the innate and adaptive immune responses triggered by metal-based nanoparticles in a variety of immune system models. Yueh-Hsia Luo, Louis W. Chang, and Pinpin Lin Copyright © 2015 Yueh-Hsia Luo et al. All rights reserved. Aquatic Toxicity Comparison of Silver Nanoparticles and Silver Nanowires Mon, 01 Jun 2015 07:27:17 +0000 To better understand the potential ecotoxicological impact of silver nanoparticles (AgNPs) and silver nanowires (AgNWs) released into freshwater environments, the toxicities of these nanomaterials were assessed and compared using Organization for Economic Cooperation and Development (OECD) test guidelines, including a “Daphnia sp., acute immobilization test,” “Fish, acute toxicity test,” and “freshwater alga and cyanobacteria, growth inhibition test.” Based on the estimated median lethal/effective concentrations of AgNPs and AgNWs, the susceptibility to the nanomaterials was different among test organisms (daphnia > algae > fish), suggesting that the AgNPs are classified as “category acute 1” for Daphnia magna, “category acute 2” for Oryzias latipes, and “category acute 1” for Raphidocelis subcapitata, while the AgNWs are classified as “category acute 1” for Daphnia magna, “category acute 2” for Oryzias latipes, and “category acute 2” for Raphidocelis subcapitata, according to the GHS (Globally Harmonized System of Classification and Labelling of Chemicals). In conclusion, the present results suggest that more attention should be paid to prevent the accidental or intentional release of silver nanomaterials into freshwater aquatic environments. Eun Kyung Sohn, Seyed Ali Johari, Tae Gyu Kim, Jin Kwon Kim, Ellen Kim, Ji Hyun Lee, Young Shin Chung, and Il Je Yu Copyright © 2015 Eun Kyung Sohn et al. All rights reserved. Three-Day Continuous Exposure Monitoring of CNT Manufacturing Workplaces Mon, 01 Jun 2015 06:57:37 +0000 Continuous monitoring for possible exposure to carbon nanotubes was conducted over a period of 2 to 3 days at workplaces that manufacture multiwall carbon nanotubes (MWCNTs) and single wall carbon nanotubes (SWCNTs). To estimate the potential emission of carbon nanotubes (CNTs) and potential exposure of workers, personal sampling, area monitoring, and real-time monitoring using an scanning mobility particle sizer (SMPS) and dust monitor were conducted at workplaces where the workers manufactured CNTs. The personal and area sampling of the total suspended particulate (TSP) at the MWCNT manufacturing facilities ranged from 0.031 to 0.254 and from N.D (not detected) to 0.253 mg/m3, respectively. This 2- to 3-day monitoring study found that nanoparticles were released when opening the chemical vapor deposit (CVD) reactor door after the synthesis of MWCNTs, when transferring the MWCNTs to containers and during blending and grinding. However, distinguishing the background concentration from the work process particle emission was complicated due to sustained and even increased particle concentrations after the work processes were terminated. The MWCNTs sampled for transmission electron microscopy (TEM) observation exhibited a tangled shape with no individual dispersed CNT structures. Ji Hyun Lee, Kang Ho Ahn, Sun Man Kim, Ellen Kim, Gun Ho Lee, Jeong Hee Han, and Il Je Yu Copyright © 2015 Ji Hyun Lee et al. All rights reserved. Subchronic Arsenic Exposure Induces Anxiety-Like Behaviors in Normal Mice and Enhances Depression-Like Behaviors in the Chemically Induced Mouse Model of Depression Sun, 31 May 2015 07:01:46 +0000 Accumulating evidence implicates that subchronic arsenic exposure causes cerebral neurodegeneration leading to behavioral disturbances relevant to psychiatric disorders. However, there is still little information regarding the influence of subchronic exposure to arsenic-contaminated drinking water on mood disorders and its underlying mechanisms in the cerebral prefrontal cortex. The aim of this study is to assess the effects of subchronic arsenic exposure (10 mg/LAs2O3 in drinking water) on the anxiety- and depression-like behaviors in normal mice and in the chemically induced mouse model of depression by reserpine pretreatment. Our findings demonstrated that 4 weeks of arsenic exposure enhance anxiety-like behaviors on elevated plus maze (EPM) and open field test (OFT) in normal mice, and 8 weeks of arsenic exposure augment depression-like behaviors on tail suspension test (TST) and forced swimming test (FST) in the reserpine pretreated mice. In summary, in this present study, we demonstrated that subchronic arsenic exposure induces only the anxiety-like behaviors in normal mice and enhances the depression-like behaviors in the reserpine induced mouse model of depression, in which the cerebral prefrontal cortex BDNF-TrkB signaling pathway is involved. We also found that eight weeks of subchronic arsenic exposure are needed to enhance the depression-like behaviors in the mouse model of depression. These findings imply that arsenic could be an enhancer of depressive symptoms for those patients who already had the attribute of depression. Chia-Yu Chang, How-Ran Guo, Wan-Chen Tsai, Kai-Lin Yang, Li-Chuan Lin, Tain-Junn Cheng, and Jiunn-Jye Chuu Copyright © 2015 Chia-Yu Chang et al. All rights reserved. Effects of Atrazine on the Development of Neural System of Zebrafish, Danio rerio Sun, 31 May 2015 06:55:56 +0000 By comparative analysis of histomorphology and AChE activity, the changes of physiological and biochemical parameters were determined in zebrafish embryos and larvae dealt with atrazine (ATR) at different concentrations (0.0001, 0.001, 0.01, 0.1, and 1 mg/L). This study showed that the development of the sarcomere and the arrangement of white muscle myofibers were affected by ATR significantly and the length of sarcomere shortened. Further analysis of the results showed that the AChE activity in juvenile fish which was treated with ATR was downregulated, which can indicate that the innervation efficiency to the muscle was impaired. Conversely, the AChE activity in zebrafish embryos which was treated with ATR was upregulated. A parallel phenomenon showed that embryonic primary sensory neurons (Rohon-Beard cells), principally expressing AChE in embryos, survived the physiological apoptosis. These phenomena demonstrated that the motor integration ability of the zebrafish was damaged by ATR which can disturb the development of sensory neurons and sarcomere and the innervations of muscle. Hao Wang, Shumei Mu, Fengjuan Zhang, Hailing Wang, Huan Liu, Han Zhang, and Xianjiang Kang Copyright © 2015 Hao Wang et al. All rights reserved. Toxicity of Nanomaterials Wed, 20 May 2015 14:14:38 +0000 Haseeb A. Khan and Rishi Shanker Copyright © 2015 Haseeb A. Khan and Rishi Shanker. All rights reserved. Effects of Two Fullerene Derivatives on Monocytes and Macrophages Tue, 19 May 2015 09:35:55 +0000 Two fullerene derivatives (fullerenes 1 and 2), bearing a hydrophilic chain on the pyrrolidinic nitrogen, were developed with the aim to deliver anticancer agents to solid tumors. These two compounds showed a significantly different behaviour on human neoplastic cell lines in vitro in respect to healthy leukocytes. In particular, the pyrrolidinium ring on the fullerene carbon cage brings to a more active compound. In the present work, we describe the effects of these fullerenes on primary cultures of human monocytes and macrophages, two kinds of immune cells representing the first line of defence in the immune response to foreign materials. These compounds are not recognized by circulating monocytes while they get into macrophages. The evaluation of the pronecrotic or proapoptotic effects, analysed by means of analysis of the purinergic receptor P2X7 activation and of ROS scavenging activity, has allowed us to show that fullerene 2, but not its analogue fullerene 1, displays toxicity, even though at concentrations higher than those shown to be active on neoplastic cells. Sabrina Pacor, Alberto Grillo, Luka Đorđević, Sonia Zorzet, Marianna Lucafò, Tatiana Da Ros, Maurizio Prato, and Gianni Sava Copyright © 2015 Sabrina Pacor et al. All rights reserved. Bucky Tubes Induce Oxidative Stress Mediated Cell Death in Human Lung Cells Mon, 18 May 2015 07:22:34 +0000 Unique physicochemical properties of carbon nanomaterials (CNMs) have opened a new era for therapeutics and diagnosis (known as theranostics) of various diseases. This exponential increase in application makes them important for toxicology studies. The present study was aimed at exploring the toxic potential of one of the CNMs, that is, bucky tubes (BTs), in human lung adenocarcinoma (A549) cell line. BTs were characterised by electron microscopy (TEM), dynamic light scattering (DLS), Fourier transform spectroscopy (FTIR), and X-ray diffraction (XRD). Flow cytometric study showed a concentration and time dependent increase in intracellular internalization as well as reduction in cell viability upon exposure to BTs. However, a significant increase in intracellular reactive oxygen species (ROS) production was observed as evident by increased fluorescence intensity of 2′,7′-dichlorofluorescein (DCF). BTs induced oxidative stress in cells as evident by depletion in glutathione with concomitant increase in lipid peroxidation with increasing concentrations. A significant increase in micronucleus formation and apoptotic cell population and loss of mitochondrial membrane potential (MMP) as compared to control were observed. Moreover, in the present study, BTs were found to be mild toxic and it is encouraging to conclude that BTs having outer diameter in the range of 7–12 nm and length 0.5–10 μm can be used for theranostics. Jaya Singhal, Surinder P. Singh, Stalin Karuppiah, and Alok K. Pandey Copyright © 2015 Jaya Singhal et al. All rights reserved. Endocytosis of Multiwalled Carbon Nanotubes in Bronchial Epithelial and Mesothelial Cells Mon, 18 May 2015 07:06:53 +0000 Bronchial epithelial cells and mesothelial cells are crucial targets for the safety assessment of inhalation of carbon nanotubes (CNTs), which resemble asbestos particles in shape. Intrinsic properties of multiwalled CNTs (MWCNTs) are known to cause potentially hazardous effects on intracellular and extracellular pathways. These interactions alter cellular signaling and affect major cell functions, resulting in cell death, lysosome injury, reactive oxygen species production, apoptosis, and cytokine release. Furthermore, CNTs are emerging as a novel class of autophagy inducers. Thus, in this study, we focused on the mechanisms of MWCNT uptake into the human bronchial epithelial cells (HBECs) and human mesothelial cells (HMCs). We verified that MWCNTs are actively internalized into HBECs and HMCs and were accumulated in the lysosomes of the cells after 24-hour treatment. Next, we determined which endocytosis pathways (clathrin-mediated, caveolae-mediated, and macropinocytosis) were associated with MWCNT internalization by using corresponding endocytosis inhibitors, in two nonphagocytic cell lines derived from bronchial epithelial cells and mesothelioma cells. Clathrin-mediated endocytosis inhibitors significantly suppressed MWCNT uptake, whereas caveolae-mediated endocytosis and macropinocytosis were also found to be involved in MWCNT uptake. Thus, MWCNTs were positively taken up by nonphagocytic cells, and their cytotoxicity was closely related to these three endocytosis pathways. Kayo Maruyama, Hisao Haniu, Naoto Saito, Yoshikazu Matsuda, Tamotsu Tsukahara, Shinsuke Kobayashi, Manabu Tanaka, Kaoru Aoki, Seiji Takanashi, Masanori Okamoto, and Hiroyuki Kato Copyright © 2015 Kayo Maruyama et al. All rights reserved. Nanotoxic Profiling of Novel Iron Oxide Nanoparticles Functionalized with Perchloric Acid and SiPEG as a Radiographic Contrast Medium Sun, 17 May 2015 14:16:11 +0000 Emerging syntheses and findings of new metallic nanoparticles (MNPs) have become an important aspect in various fields including diagnostic imaging. To date, iodine has been utilized as a radiographic contrast medium. However, the raise concern of iodine threats on iodine-intolerance patient has led to search of new contrast media with lower toxic level. In this animal modeling study, 14 nm iron oxide nanoparticles (IONPs) with silane-polyethylene glycol (SiPEG) and perchloric acid have been assessed for toxicity level as compared to conventional iodine. The nanotoxicity of IONPs was evaluated in liver biochemistry, reactive oxygen species production (ROS), lipid peroxidation mechanism, and ultrastructural evaluation using transmission electron microscope (TEM). The hematological analysis and liver function test (LFT) revealed that most of the liver enzymes were significantly higher in iodine-administered group as compared to those in normal and IONPs groups . ROS production assay and lipid peroxidation indicator, malondialdehyde (MDA), also showed significant reductions in comparison with iodine group . TEM evaluation yielded the aberration of nucleus structure of iodine-administered group as compared to those in control and IONPs groups. This study has demonstrated the less toxic properties of IONPs and it may postulate that IONPs are safe to be applied as radiographic contrast medium. Muhamad Idham Mohamed, Mohd Khairul Amran Mohammad, Hairil Rashmizal Abdul Razak, Khairunisak Abdul Razak, and Wan Mazlina Md Saad Copyright © 2015 Muhamad Idham Mohamed et al. All rights reserved. The Janus Facet of Nanomaterials Sun, 17 May 2015 14:06:29 +0000 Application of nanoscale materials (NMs) displays a rapidly increasing trend in electronics, optics, chemical catalysis, biotechnology, and medicine due to versatile nature of NMs and easily adjustable physical, physicochemical, and chemical properties. However, the increasing abundance of NMs also poses significant new and emerging health and environmental risks. Despite growing efforts, understanding toxicity of NMs does not seem to cope with the demand, because NMs usually act entirely different from those of conventional small molecule drugs. Currently, large-scale application of available safety assessment protocols, as well as their furthering through case-by-case practice, is advisable. We define a standard work-scheme for nanotoxicity evaluation of NMs, comprising thorough characterization of structural, physical, physicochemical, and chemical traits, followed by measuring biodistribution in live tissue and blood combined with investigation of organ-specific effects especially regarding the function of the brain and the liver. We propose a range of biochemical, cellular, and immunological processes to be explored in order to provide information on the early effects of NMs on some basic physiological functions and chemical defense mechanisms. Together, these contributions give an overview with important implications for the understanding of many aspects of nanotoxicity. Julianna Kardos, Katalin Jemnitz, István Jablonkai, Attila Bóta, Zoltán Varga, Júlia Visy, and László Héja Copyright © 2015 Julianna Kardos et al. All rights reserved. Cytotoxicity of Protein-Carbon Nanotubes on J774 Macrophages Is a Functionalization Grade-Dependent Effect Sun, 17 May 2015 13:16:59 +0000 Carbon nanotubes (CNTs) are used as carriers in medicine due to their ability to be functionalized with chemical substances. However, cytotoxicity analysis is required prior to use for in vivo models. The aim of this study was to evaluate the cytotoxic effect of CNTs functionalized with a 46 kDa surface protein from Entamoeba histolytica (P46-CNTs) on J774A macrophages. With this purpose, CNTs were synthesized by spray pyrolysis and purified (P-CNTs) using sonication for 48 h. A 46 kDa protein, with a 4.6–5.4 pI range, was isolated from E. histolytica HM1:IMSS strain trophozoites using an OFFGEL system. The P-CNTs were functionalized with the purified 46 kDa protein, classified according to their degree of functionalization, and characterized by Raman and Infrared spectroscopy. In vitro cytotoxicity was evaluated by MTT, apoptosis, and morphological assays. The results demonstrated that P46-CNTs exhibited cytotoxicity dependent upon the functionalized grade. Contrary to what was expected, P46-CNTs with a high grade of functionalization were more toxic to J774 macrophages than P46-CNTs with a low grade of functionalization, than P-CNTs, and had a similar level of toxicity as UP-CNT. This suggests that the nature of the functionalized protein plays a key role in the cytotoxicity of these nanoparticles. Silvia Lorena Montes-Fonseca, Blanca Sánchez-Ramírez, Antonia Luna-Velasco, Carlos Arzate-Quintana, Macrina Beatriz Silva-Cazares, Carmen González Horta, and Erasmo Orrantia-Borunda Copyright © 2015 Silvia Lorena Montes-Fonseca et al. All rights reserved. Toxic Effects of Nickel Oxide Bulk and Nanoparticles on the Aquatic Plant Lemna gibba L. Sun, 17 May 2015 13:05:26 +0000 The aquatic plant Lemna gibba L. was used to investigate and compare the toxicity induced by 30 nm nickel oxide nanoparticles (NiO-NPs) and nickel(II) oxide as bulk (NiO-Bulk). Plants were exposed during 24 h to 0–1000 mg/L of NiO-NPs or NiO-Bulk. Analysis of physicochemical characteristics of nanoparticles in solution indicated agglomerations of NiO-NPs in culture medium and a wide size distribution was observed. Both NiO-NPs and NiO-Bulk caused a strong increase in reactive oxygen species (ROS) formation, especially at high concentration (1000 mg/L). These results showed a strong evidence of a cellular oxidative stress induction caused by the exposure to NiO. Under this condition, NiO-NPs and NiO-Bulk induced a strong inhibitory effect on the PSII quantum yield, indicating an alteration of the photosynthetic electron transport performance. Under the experimental conditions used, it is clear that the observed toxicity impact was mainly due to NiO particles effect. Therefore, results of this study permitted determining the use of ROS production as an early biomarker of NiO exposure on the aquatic plant model L. gibba used in toxicity testing. Abdallah Oukarroum, Lotfi Barhoumi, Mahshid Samadani, and David Dewez Copyright © 2015 Abdallah Oukarroum et al. All rights reserved. Toxicity of Graphene Shells, Graphene Oxide, and Graphene Oxide Paper Evaluated with Escherichia coli Biotests Sun, 17 May 2015 12:57:22 +0000 The plate-like graphene shells (GS) produced by an original methane pyrolysis method and their derivatives graphene oxide (GO) and graphene oxide paper (GO-P) were evaluated with luminescent Escherichia coli biotests and additional bacterial-based assays which together revealed the graphene-family nanomaterials’ toxicity and bioactivity mechanisms. Bioluminescence inhibition assay, fluorescent two-component staining to evaluate cell membrane permeability, and atomic force microscopy data showed GO expressed bioactivity in aqueous suspension, whereas GS suspensions and the GO-P surface were assessed as nontoxic materials. The mechanism of toxicity of GO was shown not to be associated with oxidative stress in the targeted soxS::lux and katG::lux reporter cells; also, GO did not lead to significant mechanical disruption of treated bacteria with the release of intracellular DNA contents into the environment. The well-coordinated time- and dose-dependent surface charge neutralization and transport and energetic disorders in the Escherichia coli cells suggest direct membrane interaction, internalization, and perturbation (i.e., “membrane stress”) as a clue to graphene oxide’s mechanism of toxicity. Ludmila V. Efremova, Alexey S. Vasilchenko, Eduard G. Rakov, and Dmitry G. Deryabin Copyright © 2015 Ludmila V. Efremova et al. All rights reserved. Estrogenic Evaluation and Organochlorine Identification in Blubber of North Sea Harbour Porpoise (Phocoena phocoena) Stranded on the North Sea Coast Sun, 17 May 2015 09:13:45 +0000 Thirteen individual organochlorine compounds at 3 concentrations (80, 400, and 2000 ng/mL culture medium), as well as mixtures, were assayed for the estrogen receptor (ER) activation or inhibition, using a luciferase reporter gene assay (RGA). None of the PCB 138, 153, or 180 or their mixture induced a response in the RGA. o,p′-DDT was the most potent xenoestrogen from the DDT group, inducing a response already at 80 ng/mL. From the HCH and HCB group, only β-HCH (at 400 and 2000 ng/mL) and δ-HCH (at 2000 ng/mL) displayed estrogenic activities. These 13 organochlorines were determined by GC-MS in 12 samples of North Sea harbor porpoise blubber. The PCBs were the main contaminants. Within each group, PCB 153 (6.0 × 102~4.2 × 104 μg/kg), p,p′-DDE (5.1 × 102~8.6 × 103 μg/kg), and HCB (7.6 × 101~1.5 × 103 μg/kg) were the compounds found in highest concentrations. The hormonal activity of the porpoise blubber samples was also assayed in RGA, where two samples showed estrogenic activity, seven samples showed antiestrogenic activity, and one sample showed both estrogenic and antiestrogenic activity. Our results suggest that the 13 POPs measured by GC-MS in the samples cannot explain alone the estrogenicity of the extracts. Pedro Henrique Imazaki, François Brose, Thierry Jauniaux, Krishna Das, Marc Muller, and Marie-Louise Scippo Copyright © 2015 Pedro Henrique Imazaki et al. All rights reserved. Unsaturated Fatty Acids Supplementation Reduces Blood Lead Level in Rats Thu, 14 May 2015 07:25:25 +0000 Some dietary factors could inhibit lead toxicity. The aim of this study was to evaluate the effect of dietary compounds rich in unsaturated fatty acids (FA) on blood lead level, lipid metabolism, and vascular reactivity in rats. Serum metallothionein and organs’ lead level were evaluated with the aim of assessing the possible mechanism of unsaturated FA impact on blood lead level. For three months, male Wistar rats that were receiving drinking water with (100 ppm Pb) or without lead acetate were supplemented per os daily with virgin olive oil or linseed oil (0.2 mL/kg b.w.) or egg derived lecithin fraction: “super lecithin” (50 g/kg b.w.). Mesenteric artery was stimulated ex vivo by norepinephrine (NE) administered at six different doses. Lecithin supplementation slightly reduced pressor responses of artery to NE. Lead administered to rats attenuated the beneficial effect of unsaturated FA on lipid metabolism and vascular reactivity to adrenergic stimulation. On the other hand, the super lecithin and linseed oil that were characterized by low omega-6 to omega-3 ratio (about 1) reduced the blood lead concentration. This effect was observed in lead poisoned rats and also in rats nonpoisoned with lead . Anna Skoczyńska, Anna Wojakowska, Dorian Nowacki, Łukasz Bobak, Barbara Turczyn, Beata Smyk, Andrzej Szuba, and Tadeusz Trziszka Copyright © 2015 Anna Skoczyńska et al. All rights reserved. In Vitro and In Vivo Cytogenotoxic Effects of Hot Aqueous Extract of Achyrocline satureioides (Lam.) DC. Mon, 11 May 2015 11:21:25 +0000 In this work we extend the toxicological studies of hot aqueous extract of A. satureioides (As-HAE) evaluating cytotoxic and apoptotic effects on human peripheral blood mononuclear cells (PBMCs). We also determine genotoxic action of this extract in vivo. In addition, the extract was chemically characterized. Finally, we established a comparison with previous data of cold aqueous extract. The As-HAE induced cytotoxicity on PBMCs determined by trypan blue dye exclusion (CC50 = 653 μg/mL) and MTT (CC50 = 588 μg/mL) assays being more toxic than cold extract. However, As-HAE as well as cold extract did not induce apoptosis measured by Hoechst 33258 staining, TUNEL assay, and DNA fragmentation analysis. The in vivo micronucleus test showed that As-HAE exerted cytogenotoxic effects on bone marrow of mice, contrary to what was observed with cold extract. The chemical study of As-HAE allowed identifying the flavonoids found in cold extract: luteolin, quercetin, and 3-O-methylquercetin, but at higher concentrations. We suggest that toxic effects induced by As-HAE could be due to high concentrations of these flavonoids. Given that As-HAE is the most used in folkloric medicine, its administration should be controlled in order to prevent potential cell damage. L. N. Cariddi, M. C. Sabini, F. M. Escobar, R. Bacchetti, I. Montironi, C. Merckis, E. B. Reinoso, S. Núñez Montoya, S. M. Zanon, L. R. Comini, and L. I. Sabini Copyright © 2015 L. N. Cariddi et al. All rights reserved. Hypocholesterolemic Effect and In Vitro Pancreatic Lipase Inhibitory Activity of an Opuntia ficus-indica Extract Mon, 11 May 2015 11:12:38 +0000 Cholesterol control is fundamental for prevention of cardiovascular disorders. In this work, the hypocholesterolemic activity of an aqueous Opuntia ficus-indica extract (AOE) was tested in triton-induced mice. The inhibitory activity on pancreatic lipase enzyme was evaluated in vitro by the same extract. Furthermore, polyphenol content of the extract was evaluated. Hypercholesterolemia was induced in three groups of mice by intraperitoneal administration of Triton WR-1339. After induction of hypercholesterolemia, the groups were treated with an AOE (500 mg/kg) and saline solution and the positive control group with orlistat, respectively. Cholesterol levels were measured 24 h later in peripheral blood. The levels of blood cholesterol after administration of AOE significantly decreased compared to negative control. The inhibitory activity of AOE on pancreatic lipase enzyme was evaluated at concentrations from 60 to 1000 μg/mL. The AOE inhibited the pancreatic lipase with an IC50 = 588.5 μg/mL. The AOE had a high content of polyphenolic compounds. These results show that AOE is able to prevent hypercholesterolemia by pancreatic lipase inhibition, in part due to its polyphenolic compounds. Eduardo Padilla-Camberos, Jose Miguel Flores-Fernandez, Ofelia Fernandez-Flores, Yanet Gutierrez-Mercado, Joel Carmona-de la Luz, Fabiola Sandoval-Salas, Carlos Mendez-Carreto, and Kirk Allen Copyright © 2015 Eduardo Padilla-Camberos et al. All rights reserved. Transcriptome Analysis of the Carmine Spider Mite, Tetranychus cinnabarinus (Boisduval, 1867) (Acari: Tetranychidae), and Its Response to β-Sitosterol Mon, 11 May 2015 10:35:27 +0000 Tetranychus cinnabarinus (Acari: Tetranychidae) is a worldwide polyphagous agricultural pest that has the title of resistance champion among arthropods. We reported previously the identification of the acaricidal compound β-sitosterol from Mentha piperita and Inula japonica. However, the acaricidal mechanism of β-sitosterol is unclear. Due to the limited genetic research carried out, we de novo assembled the transcriptome of T. cinnabarinus using Illumina sequencing and conducted a differential expression analysis of control and β-sitosterol-treated mites. In total, we obtained >5.4 G high-quality bases for each sample with unprecedented sequencing depth and assembled them into 22,941 unigenes. We identified 617 xenobiotic metabolism-related genes involved in detoxification, binding, and transporting of xenobiotics. A highly expanded xenobiotic metabolic system was found in mites. T. cinnabarinus detoxification genes—including carboxyl/cholinesterase and ABC transporter class C—were upregulated after β-sitosterol treatment. Defense-related proteins, such as Toll-like receptor, legumain, and serine proteases, were also activated. Furthermore, other important genes—such as the chloride channel protein, cytochrome b, carboxypeptidase, peritrophic membrane chitin binding protein, and calphostin—may also play important roles in mites’ response to β-sitosterol. Our results demonstrate that high-throughput-omics tool facilitates identification of xenobiotic metabolism-related genes and illustration of the acaricidal mechanisms of β-sitosterol. Chunya Bu, Jinling Li, Xiao-Qin Wang, Guanglu Shi, Bo Peng, Jingyu Han, Pin Gao, and Younian Wang Copyright © 2015 Chunya Bu et al. All rights reserved. Effect of Metformin on Viability, Morphology, and Ultrastructure of Mouse Bone Marrow-Derived Multipotent Mesenchymal Stromal Cells and Balb/3T3 Embryonic Fibroblast Cell Line Thu, 30 Apr 2015 11:54:22 +0000 Metformin, a popular drug used to treat diabetes, has recently gained attention as a potentially useful therapeutic agent for treating cancer. In our research metformin was added to in vitro cultures of bone marrow-derived multipotent mesenchymal stromal cells (BMSCs) and Balb/3T3 fibroblast at concentration of 1 mM, 5 mM, and 10 mM. Obtained results indicated that metformin negatively affected proliferation activity of investigated cells. The drug triggered the formation of autophagosomes and apoptotic bodies in all tested cultures. Additionally, we focused on determination of expression of genes involved in insulin-like growth factor 2 (IGF2) signaling pathway. The most striking finding was that the mRNA level of IGF2 was constant in both BMSCs and Balb/3T3. Further, the analysis of IGF2 concentration in cell supernatants showed that it decreased in BMSC cultures after 5 and 10 mM metformin treatments. In case of Balb/3T3 the concentration of IGF2 in culture supernatants decreased after 1 and 5 mM and increased after 10 mM of metformin. Our results suggest that metformin influences the cytophysiology of somatic cells in a dose- and time-dependent manner causing inhibition of proliferation and abnormalities of their morphology and ultrastructure. Agnieszka Śmieszek, Aleksandra Czyrek, Katarzyna Basinska, Justyna Trynda, Aneta Skaradzińska, Anna Siudzińska, Monika Marędziak, and Krzysztof Marycz Copyright © 2015 Agnieszka Śmieszek et al. All rights reserved. Environmental Toxicology in Addressing Public Health Challenges in East Asia Wed, 29 Apr 2015 13:15:34 +0000 How-Ran Guo, Zailina Hashim, Shih-Bin Su, and Jochen Bundschuh Copyright © 2015 How-Ran Guo et al. All rights reserved. Decreased Pulmonary Function in School Children in Western Japan after Exposures to Asian Desert Dusts and Its Association with Interleukin-8 Tue, 28 Apr 2015 06:57:11 +0000 The objective of the study was to investigate the influence of Asian dust storms (ADS) on pulmonary function of school children and the relationship of this effect with interleukin-8. Morning peak expiratory flow (PEF) was measured daily in 399 children from April to May 2012 and in 384 of these children from March to May 2013. The data were analyzed for an association between ADS events and PEF by linear mixed models. Interleukin-8 transcriptional activity was assessed in THP-G8 cells stimulated by airborne particles collected on ADS days. Seven ADS days were identified: April 23 and 24, 2012; March 8 to 10, 2013; and March 19 and 20, 2013. Changes in PEF after ADS exposure were −8.17 L/min (95% confidence interval, −11.40 to −4.93) in 2012 and −1.17 L/min (−4.07 to 1.74) in 2013, and there was a significant difference between 2012 and 2013. Interleukin-8 transcriptional activity was significantly higher in 2012 at -fold compared to in March 8 to 10, 2013, and in March 19 and 20, 2013. The influence of ADS events on pulmonary function of children differs with each ADS event and may be related to interleukin-8 production. Masanari Watanabe, Hisashi Noma, Jun Kurai, Hiroyuki Sano, Rumiko Saito, Satoshi Abe, Yutaka Kimura, Setsuya Aiba, Mitsuo Oshimura, Akira Yamasaki, and Eiji Shimizu Copyright © 2015 Masanari Watanabe et al. All rights reserved. Association between Arsenic Exposure and Diabetes: A Meta-Analysis Mon, 27 Apr 2015 08:01:23 +0000 Studies on the association between arsenic exposure and diabetes mellitus (DM) yielded inconsistent results. Epidemiologic data on the associations between arsenic exposures via inhalation and DM are limited. Therefore, we conducted a meta-analysis to evaluate the risk of DM associated with arsenic exposure. We searched the related literature through a systematic approach and analyzed the data according to the exposure route (inhalation and ingestion). We used random-effect models to estimate the summary relative risks (RRs) for DM associated with arsenic exposure and used I2 statistics to assess the heterogeneity of studies. We identified 38 relevant studies, of which the 32 on the ingestion route showed a significant association between arsenic exposure and DM (RR = 1.57; 95% CI 1.27–1.93). Focusing on the 24 studies in which the diagnosis of DM was confirmed using laboratory tests or medical records, we found that the summary RR was 1.71 (95% CI 1.32–2.23), very close to the overall estimates. We concluded that ingested arsenic is associated with the development of DM, but the heterogeneity among the studies may affect the results. Tzu-Ching Sung, Jhih-Wei Huang, and How-Ran Guo Copyright © 2015 Tzu-Ching Sung et al. All rights reserved. Indoor Air Quality and Respiratory Health among Malay Preschool Children in Selangor Thu, 23 Apr 2015 12:41:22 +0000 Indoor air quality (IAQ) has been the object of several studies due to its adverse health effects on children. Methods. A cross-sectional comparative study was carried out among Malay children in Balakong (2 studied preschools) and Bangi (2 comparative preschools), Selangor, with the aims of determining IAQ and its association with respiratory health. 61 and 50 children aged 5-6 years were selected as studied and comparative groups. A questionnaire was used to obtain an exposure history and respiratory symptoms. Lung function test was carried out. IAQ parameters obtained include indoor concentration of particulate matter (PM), volatile organic compounds (VOCs), carbon monoxide (CO), carbon dioxide (CO2), temperature, air velocity (AV), and relative humidity. Results. There was a significant difference between IAQ in studied and comparative preschools for all parameters measured except for CO2 and AV. Studied preschools had higher PM and CO concentration. FVC, FEV1, FVC% and FEV1% predicted values were significantly lower among studied group. Exposures to PM, VOCs, and CO were associated with wheezing. Conclusion. The finding concluded that exposures to poor IAQ might increase the risk of getting lung function abnormality and respiratory problems among study respondents. Nur Azwani Mohd Nor Rawi, Juliana Jalaludin, and Poh Choo Chua Copyright © 2015 Nur Azwani Mohd Nor Rawi et al. All rights reserved. Biomarker as a Research Tool in Linking Exposure to Air Particles and Respiratory Health Thu, 23 Apr 2015 12:33:49 +0000 Some of the environmental toxicants from air pollution include particulate matter (PM10), fine particulate matter (PM2.5), and ultrafine particles (UFP). Both short- and long-term exposure could result in various degrees of respiratory health outcomes among exposed persons, which rely on the individuals’ health status. Methods. In this paper, we highlight a review of the studies that have used biomarkers to understand the association between air particles exposure and the development of respiratory problems resulting from the damage in the respiratory system. Data from previous epidemiological studies relevant to the application of biomarkers in respiratory system damage reported from exposure to air particles are also summarized. Results. Based on these analyses, the findings agree with the hypothesis that biomarkers are relevant in linking harmful air particles concentrations to increased respiratory health effects. Biomarkers are used in epidemiological studies to provide an understanding of the mechanisms that follow airborne particles exposure in the airway. However, application of biomarkers in epidemiological studies of health effects caused by air particles in both environmental and occupational health is inchoate. Conclusion. Biomarkers unravel the complexity of the connection between exposure to air particles and respiratory health. Nur Faseeha Suhaimi and Juliana Jalaludin Copyright © 2015 Nur Faseeha Suhaimi and Juliana Jalaludin. All rights reserved. Corrigendum to “Synergistic Effects of Toxic Elements on Heat Shock Proteins” Thu, 23 Apr 2015 06:51:45 +0000 Khalid Mahmood, Saima Jadoon, Qaisar Mahmood, Muhammad Irshad, and Jamshaid Hussain Copyright © 2015 Khalid Mahmood et al. All rights reserved. Histopathological, Ultrastructural, and Immunohistochemical Assessment of Hippocampus Structures of Rats Exposed to TCDD and High Doses of Tocopherol and Acetylsalicylic Acid Tue, 24 Mar 2015 09:53:04 +0000 The effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on central nervous system consists of changing expression of estrogen receptors, whereas the result of chronic inflammatory reaction caused by dioxin is occurrence of destructive changes in various organs connected with disturbed metabolism of connective tissue and damage of cells. The aim of the study was to determine the effect of dioxins on function, ultrastructure, and cytological and histological structure of hippocampus, particularly on expression of estrogen receptors in central nervous system as well as to define protective influence of tocopherol (TCP) and acetylsalicylic acid (ASA) on the decrease in activity of proinflammatory effects in central nervous system. It was shown that TCDD contributes to destructive and inflammatory changes along with demyelization of myelin sheaths and atrophy of estrogen receptors in hippocampus. Dioxin contributes to atrophy of estrogen receptors in hippocampus, in which also destructive and inflammatory changes were found along with demyelination of myelin sheaths. Histopathological and ultrastructural image of hippocampus areas in rats, in which both TCP and ASA were used, is characterized by poorly expressed degenerative changes and smaller inflammatory reactivity. Using both TCP and ASA has a protective effect on functions of central nervous system. Joanna Rosińczuk, Robert Dymarek, and Ireneusz Całkosiński Copyright © 2015 Joanna Rosińczuk et al. All rights reserved. Acute and Chronic Toxicity, Cytochrome P450 Enzyme Inhibition, and hERG Channel Blockade Studies with a Polyherbal, Ayurvedic Formulation for Inflammation Tue, 17 Mar 2015 09:03:58 +0000 Ayurvedic plants are known for thousands of years to have anti-inflammatory and antiarthritic effect. We have recently shown that BV-9238, a proprietary formulation of Withania somnifera, Boswellia serrata, Zingiber officinale, and Curcuma longa, inhibits LPS-induced TNF-alpha and nitric oxide production from mouse macrophage and reduces inflammation in different animal models. To evaluate the safety parameters of BV-9238, we conducted a cytotoxicity study in RAW 264.7 cells (0.005–1 mg/mL) by MTT/formazan method, an acute single dose (2–10 g/kg bodyweight) toxicity study and a 180-day chronic study with 1 g and 2 g/kg bodyweight in Sprague Dawley rats. Some sedation, ptosis, and ataxia were observed for first 15–20 min in very high acute doses and hence not used for further chronic studies. At the end of 180 days, gross and histopathology, blood cell counts, liver and renal functions were all at normal levels. Further, a modest attempt was made to assess the effects of BV-9238 (0.5 µg/mL) on six major human cytochrome P450 enzymes and 3H radioligand binding assay with human hERG receptors. BV-9238 did not show any significant inhibition of these enzymes at the tested dose. All these suggest that BV-9238 has potential as a safe and well tolerated anti-inflammatory formulation for future use. Debendranath Dey, Sunetra Chaskar, Nitin Athavale, and Deepa Chitre Copyright © 2015 Debendranath Dey et al. All rights reserved. Natural Antioxidant Betanin Protects Rats from Paraquat-Induced Acute Lung Injury Interstitial Pneumonia Thu, 12 Mar 2015 14:13:28 +0000 The effect of betanin on a rat paraquat-induced acute lung injury (ALI) model was investigated. Paraquat was injected intraperitoneally at a single dose of 20 mg/kg body weight, and betanin (25 and 100 mg/kg/d) was orally administered 3 days before and 2 days after paraquat administration. Rats were sacrificed 24 hours after the last betanin dosage, and lung tissue and bronchoalveolar lavage fluid (BALF) were collected. In rats treated only with paraquat, extensive lung injury characteristic of ALI was observed, including histological changes, elevation of lung : body weight ratio, increased lung permeability, increased lung neutrophilia infiltration, increased malondialdehyde (MDA) and myeloperoxidase (MPO) activity, reduced superoxide dismutase (SOD) activity, reduced claudin-4 and zonula occluden-1 protein levels, increased BALF interleukin (IL-1) and tumor necrosis factor (TNF)-α levels, reduced BALF IL-10 levels, and increased lung nuclear factor kappa (NF-κB) activity. In rats treated with betanin, paraquat-induced ALI was attenuated in a dose-dependent manner. In conclusion, our results indicate that betanin attenuates paraquat-induced ALI possibly via antioxidant and anti-inflammatory mechanisms. Thus, the potential for using betanin as an auxilliary therapy for ALI should be explored further. Junyan Han, Deshun Ma, Miao Zhang, Xuelian Yang, and Dehong Tan Copyright © 2015 Junyan Han et al. All rights reserved. The Mechanism of Melanocytes-Specific Cytotoxicity Induced by Phenol Compounds Having a Prooxidant Effect, relating to the Appearance of Leukoderma Thu, 12 Mar 2015 11:05:18 +0000 Specific phenol compounds including rhododendrol (RD), a skin-brightening ingredient in cosmetics, are reported to induce leukoderma, inducing a social problem, and the elucidation of mechanism of leukoderma is strongly demanded. This study investigated the relationship among the cytotoxicities of six phenol compounds on B16F10 melanoma cells and HaCaT keratinocytes and generated reactive oxygen species (ROS). As a result, the cytotoxicity of RD on B16F10 cells was higher than that on HaCaT cells, and RD significantly increased intracellular ROS and hydrogen peroxide (H2O2) levels in B16F10 cells. Furthermore, although raspberry ketone (RK), RD derivative, also increased intracellular ROS in B16F10 cells, increase in ROS was suppressed by disodium dihydrogen ethylenediaminetetraacetate dehydrate (EDTA). The amounts of increased ROS with RK in HaCaT cells without melanocyte were further increased by tyrosinase. Therefore, tyrosinase, a metalloprotein having copper, was speculated to be one of causative agents allowing phenol compounds to work as a prooxidant. Hydroxyl radical was generated by adding a mixture of tyrosinase and H2O2 to RD, and the amount of the radical was further increased by UVB, indicating that RD cytotoxicity was caused by intracellularly increased ROS, which possibly related to phenol induced prooxidants. Takeshi Nagata, Shinobu Ito, Kazuyoshi Itoga, Hideko Kanazawa, and Hitoshi Masaki Copyright © 2015 Takeshi Nagata et al. All rights reserved. Evaluation of Toxic, Cytotoxic, Mutagenic, and Antimutagenic Activities of Natural and Technical Cashew Nut Shell Liquids Using the Allium cepa and Artemia salina Bioassays Tue, 10 Mar 2015 12:29:10 +0000 The cashew nut releases a substance that is known as cashew nut shell liquid (CNSL). There are both natural (iCNSL) and technical (tCNSL) cashew nut shell liquids. This study used an Artemia salina bioassay to evaluate the toxic effects of iCNSL and tCNSL cashew nut shell liquids. It also evaluated the toxicity, cytotoxicity, and mutagenicity of CNSL and its effects on the damage induced by copper sulfate (CuSO4·5H2O) on the meristems’ root of Allium cepa. Effects of the damage induced by CuSO4·5H2O were evaluated before (pre-), during (co-), and after (post-) treatments. The iCNSL contained 94.5% anacardic acid, and the tCNSL contained 91.3% cardanol. The liquids were toxic to A. salina. Toxicity, cytotoxicity, and mutagenicity were observed with iCNSL compared with the negative control. Similarly, iCNSL failed to inhibit the toxicity and cytotoxicity of CuSO4·5H2O. The tCNSL was not toxic, cytotoxic, or mutagenic in any of the concentrations. However, the lowest iCNSL concentrations and all of the tCNSL concentrations had preventive, antimutagenic, and reparative effects on micronuclei and on chromosomal aberrations in the A. cepa. Therefore, protective, modulating, and reparative effects may be observed in the A. cepa, depending on the concentration and type of CNSL used. Aracelli de Sousa Leite, Alisson Ferreira Dantas, George Laylson da Silva Oliveira, Antonio L. Gomes Júnior, Sidney Gonçalo de Lima, Antônia Maria das Graças Lopes Citó, Rivelilson M. de Freitas, Ana Amélia de C. Melo-Cavalcante, and José Arimateia Dantas Lopes Copyright © 2015 Aracelli de Sousa Leite et al. All rights reserved. Protective and Curative Effects of the Sea Cucumber Holothuria atra Extract against DMBA-Induced Hepatorenal Diseases in Rats Mon, 02 Mar 2015 09:18:29 +0000 Oxidative stress is a common mechanism contributing to the initiation and progression of hepatic damage. Hence there is a great demand for the development of agents with potent antioxidant effect. The aim of the present study is to evaluate the efficacy of Holothuria atra extract (HaE) as an antioxidant against 7,12-dimethylbenz[a]anthracene- (DMBA-) induced hepatorenal dysfunction. Experimental animals were divided into two main groups: protective and curative. Each group was then divided into five subgroups pre- or posttreated either with distilled water (DMBA subgroups) or with HaE (200 mg/kg body weight) for seven and fourteen days. Single oral administration of DMBA (15 mg/kg body weight) to Wistar rats resulted in a significant increase in the serum liver enzymes and kidney function’s parameters. DMBA increased level of liver malondialdehyde (MDA), decreased levels of reduced glutathione (GSH), glutathione-S-transferase (GST), superoxide dismutase (SOD), and catalase (CAT) in the liver tissue, and induced liver histopathological alterations. Pre- or posttreatment with HaE orally for 14 days significantly reversed the hepatorenal alterations induced following DMBA administration. In conclusion, HaE exhibits good hepatoprotective, curative, and antioxidant potential against DMBA-induced hepatorenal dysfunction in rats that might be due to decreased free radical generation. Ahmed I. Dakrory, Sohair R. Fahmy, Amel M. Soliman, Ayman S. Mohamed, and Sayed A. M. Amer Copyright © 2015 Ahmed I. Dakrory et al. All rights reserved. Chronic Ingestion of High Dosed Phikud Navakot Extraction Induces Mesangiolysis in Rats with Alteration of AQP1 and Hsp60 Expressions Sun, 01 Mar 2015 07:53:04 +0000 Phikud Navakot (PN) is commonly used in Thai traditional medicine for alleviation of cardiovascular and cerebrovascular symptoms; however little is known about the chronic toxicity effects of the extracts from the herbs in PN. Repeated extraction doses of 10, 100, and 1,000 mg/kg/day were randomly administered to both male and female Sprague Dawley rats for 12 months. Histopathological study revealed that mesangiolysis was predominately found at the highest dose. Aquaporin 1 (AQP1) expression in the mesangiolytic glomeruli was significantly lower than in the intact glomeruli. This may be relevant to an imbalance of vascular function manifested by AQP1 alteration. In the mesangiolytic glomeruli, 60 kDa heat shock protein (Hsp60) was significantly upregulated on the endothelial lining cells of aneurysm and vascular cyst. Hsp60 increase may be related to endothelial cell damage due to its intracellular protective role. Blood urea nitrogen and creatinine levels remained within their normal range indicating well-functioning renal reserve function. In conclusion, high dosed PN may affect the endothelium leading to inability of vascular permeability and consequence to mesangiolysis. Our results suggest that only a high dose of chronic oral administration of PN is relatively toxic in association with mesangiolysis. The NOAEL was determined to be 100 mg/kg/day. Kanchana Kengkoom and Sumate Ampawong Copyright © 2015 Kanchana Kengkoom and Sumate Ampawong. All rights reserved. Environmental Lead Exposure, Catalase Gene, and Markers of Antioxidant and Oxidative Stress Relation to Hypertension: An Analysis Based on the EGAT Study Sun, 22 Feb 2015 14:36:13 +0000 Lead has been linked to the development of hypertension via oxidative stress. Catalase plays an important role in the disposal of hydrogen peroxide in erythrocyte and its activity was determined by CAT gene. The aims of this study were to investigate (1) the association between blood levels of antioxidant markers such as catalase, superoxide dismutase, glutathione, glutathione peroxidase, oxidative stress-marker (malondialdehyde), and blood lead level and (2) the influence of genetic polymorphism of CAT gene (rs769217) on change in blood pressure in general population of EGAT study project. This is a cross-sectional study of 332 normotensive, 432 prehypertensive, and 222 hypertensive male subjects. Hypertensive subjects had significantly higher blood lead level (5.28 μg/dL) compared to normotensive (4.41 μg/dL) and prehypertensive (4.55 μg/dL) subjects (). These significant findings are also found in MDA levels. Moreover, individuals with TT genotype in hypertensive group had significantly higher blood lead and MDA levels (6.06 μg/dL and 9.67 μmol/L) than those with CC genotype (5.32 μg/dL and 8.31 μmol/L, ). Our findings suggested that decreased blood catalase activity in this polymorphism together with low level lead exposure induced lipid peroxidation may be responsible for hypertension. Jintana Sirivarasai, Sukhumpun Kaojarern, Suwannee Chanprasertyothin, Pachara Panpunuan, Krittaya Petchpoung, Aninthita Tatsaneeyapant, Krongtong Yoovathaworn, Thunyachai Sura, Sming Kaojarern, and Piyamit Sritara Copyright © 2015 Jintana Sirivarasai et al. All rights reserved. Comment on “Effectiveness of Naltrexone in the Prevention of Delayed Respiratory Arrest in Opioid-Naive Methadone-Intoxicated Patients” Sun, 22 Feb 2015 14:08:36 +0000 Abbas Aghabiklooei, Hossein Hassanian-Moghaddam, and Nasim Zamani Copyright © 2015 Abbas Aghabiklooei et al. All rights reserved. Effect of Sodium Benzoate Preservative on Micronucleus Induction, Chromosome Break, and Ala40Thr Superoxide Dismutase Gene Mutation in Lymphocytes Tue, 17 Feb 2015 10:54:39 +0000 Sodium benzoate is food preservative that inhibits microbial growth. The effects of sodium benzoate preservative on micronucleus induction, chromosome break, and Ala40Thr superoxide dismutase gene mutation in lymphocytes were studied. Sodium benzoate concentrations of 0.5, 1.0, 1.5, and 2.0 mg/mL were treated in lymphocyte cell line for 24 and 48 hrs, respectively. Micronucleus test, standard chromosome culture technique, PCR, and automated sequencing technique were done to detect micronucleus, chromosome break, and gene mutation. The results showed that, at 24- and 48-hour. incubation time, sodium benzoate concentrations of 1.0, 1.5, and 2.0 mg/mL increased micronucleus formation when comparing with the control group (). At 24- and 48-hour. incubation time, sodium benzoate concentrations of 2.0 mg/mL increased chromosome break when comparing with the control group (). Sodium benzoate did not cause Ala40Thr (GCGACG) in superoxide dismutase gene. Sodium benzoate had the mutagenic and cytotoxic toxicity in lymphocytes caused by micronucleus formation and chromosome break. Malinee Pongsavee Copyright © 2015 Malinee Pongsavee. All rights reserved. DNaseI Protects against Paraquat-Induced Acute Lung Injury and Pulmonary Fibrosis Mediated by Mitochondrial DNA Wed, 11 Feb 2015 11:42:38 +0000 Background. Paraquat (PQ) poisoning is a lethal toxicological challenge that served as a disease model of acute lung injury and pulmonary fibrosis, but the mechanism is undetermined and no effective treatment has been discovered. Methods and Findings. We demonstrated that PQ injures mitochondria and leads to mtDNA release. The mtDNA mediated PBMC recruitment and stimulated the alveolar epithelial cell production of TGF-β1 in vitro. The levels of mtDNA in circulation and bronchial alveolar lavage fluid (BALF) were elevated in a mouse of PQ-induced lung injury. DNaseI could protect PQ-induced lung injury and significantly improved survival. Acute lung injury markers, such as TNFα, IL-1β, and IL-6, and marker of fibrosis, collagen I, were downregulated in parallel with the elimination of mtDNA by DNaseI. These data indicate a possible mechanism for PQ-induced, mtDNA-mediated lung injury, which may be shared by other causes of lung injury, as suggested by the same protective effect of DNaseI in bleomycin-induced lung injury model. Interestingly, increased mtDNA in the BALF of patients with amyopathic dermatomyositis-interstitial lung disease can be appreciated. Conclusions. DNaseI targeting mtDNA may be a promising approach for the treatment of PQ-induced acute lung injury and pulmonary fibrosis that merits fast tracking through clinical trials. Guo Li, Li Yuzhen, Chen Yi, Chen Xiaoxiang, Zhou Wei, Zhu Changqing, and Ye Shuang Copyright © 2015 Guo Li et al. All rights reserved. PEGylated Carbon Nanotubes Impair Retrieval of Contextual Fear Memory and Alter Oxidative Stress Parameters in the Rat Hippocampus Mon, 09 Feb 2015 13:24:31 +0000 Carbon nanotubes (CNT) are promising materials for biomedical applications, especially in the field of neuroscience; therefore, it is essential to evaluate the neurotoxicity of these nanomaterials. The present work assessed the effects of single-walled CNT functionalized with polyethylene glycol (SWCNT-PEG) on the consolidation and retrieval of contextual fear memory in rats and on oxidative stress parameters in the hippocampus. SWCNT-PEG were dispersed in water at concentrations of 0.5, 1.0, and 2.1 mg/mL and infused into the rat hippocampus. The infusion was completed immediately after training and 30 min before testing of a contextual fear conditioning task, resulting in exposure times of 24 h and 30 min, respectively. The results showed that a short exposure to SWCNT-PEG impaired fear memory retrieval and caused lipid peroxidation in the hippocampus. This response was transient and overcome by the mobilization of antioxidant defenses at 24 h. These effects occurred at low and intermediate but not high concentration of SWCNT-PEG, suggesting that the observed biological response may be related to the concentration-dependent increase in particle size in SWCNT-PEG dispersions. Lidiane Dal Bosco, Gisele E. B. Weber, Gustavo M. Parfitt, Karina Paese, Carla O. F. Gonçalves, Tiago M. Serodre, Clascídia A. Furtado, Adelina P. Santos, José M. Monserrat, and Daniela M. Barros Copyright © 2015 Lidiane Dal Bosco et al. All rights reserved. Cinnabar Induces Renal Inflammation and Fibrogenesis in Rats Thu, 05 Feb 2015 06:43:24 +0000 The purpose of this study was to investigate whether cinnabar causes renal inflammation and fibrosis in rats. Rats were dosed orally with cinnabar (1 g/kg/day) for 8 weeks or 12 weeks. The control rats were treated with solvent (5% carboxymethylcellulose solution) over the same time periods, respectively. Renal mercury (RHg), urinary mercury (UHg), serum creatinine (SCr), urine kidney injury molecule 1 (KIM-1), renal pathology, and renal mediators were examined. At both 8 weeks and 12 weeks, RHg, UHg, and urine KIM-1 were significantly higher in the cinnabar group than in the control group, although SCr was unchanged. Kidney lesions in the cinnabar-treated rats occurred mainly in the tubules and interstitium, including vacuolization, protein casts, infiltration of inflammatory cells, and slight increase in interstitial collagen. In addition, mild mesangial proliferation was observed in glomeruli. Moreover, the expression of inflammatory and fibrogenic mediators was upregulated in the cinnabar group. In conclusion, cinnabar may cause kidney damage due to the accumulation of mercury, and renal inflammation and slight fibrogenesis may occur in rats. In the clinic, the potential risk of renal injury due to the prolonged consumption of cinnabar should be considered even though the agent is relatively nontoxic. Ying Wang, Dapeng Wang, Jie Wu, Bohan Wang, Liangjun Wang, Xin Gao, Hai Huang, and Honglin Ma Copyright © 2015 Ying Wang et al. All rights reserved. Ring-Oxidative Biotransformation and Drug Interactions of Propofol in the Livers of Rats Sun, 01 Feb 2015 13:52:42 +0000 Propofol, an intravenous anesthetic agent, is widely used for inducing and maintaining anesthesia during surgical procedures and for sedating intensive care unit patients. In the clinic, rapid elimination is one of the major advantages of propofol. Meanwhile, the biotransformation and drug interactions of propofol in rat livers are still little known. In this study, we evaluated the ring-oxidative metabolism of propofol in phenobarbital-treated rat livers and possible drug interactions. Administration of phenobarbital to male Wistar rats significantly increased levels of hepatic cytochrome P450 (CYP) 2B1/2 and microsomal pentoxyresorufin O-dealkylase (PROD) activity. Analyses by high-performance liquid chromatography and liquid chromatography mass spectroscopy revealed that propofol was metabolized by phenobarbital-treated rat liver microsomes into 4-hydroxypropofol. In comparison, PROD activity and 4-hydroxy-propofol production from propofol metabolism were suppressed by orphenodrine, an inhibitor of CYP2B1/2, and a polyclonal antibody against rat CYP2B1/2 protein. Furthermore, exposure of rats to propofol did not affect the basal or phenobarbital-enhanced levels of hepatic CYP2B1/2 protein. Meanwhile, propofol decreased the dealkylation of pentoxyresorufin by phenobarbital-treated rat liver microsomes in a concentration-dependent manner. Taken together, this study shows that rat hepatic CYP2B1/2 plays a critical role in the ring-oxidative metabolism of propofol into 4-hydroxypropofol, and this anesthetic agent can inhibit CYP2B1/2 activity without affecting protein synthesis. Yu-Ting Tai, Yi-Ling Lin, Chia-Chen Chang, Yih-Giun Cherng, Ming-Jaw Don, and Ruei-Ming Chen Copyright © 2015 Yu-Ting Tai et al. All rights reserved. Status, Alert System, and Prediction of Cyanobacterial Bloom in South Korea Sun, 01 Feb 2015 09:55:32 +0000 Bloom-forming freshwater cyanobacterial genera pose a major ecological problem due to their ability to produce toxins and other bioactive compounds, which can have important implications in illnesses of humans and livestock. Cyanobacteria such as Microcystis, Anabaena, Oscillatoria, Phormidium, and Aphanizomenon species producing microcystins and anatoxin-a have been predominantly documented from most South Korean lakes and reservoirs. With the increase in frequency of such blooms, various monitoring approaches, treatment processes, and prediction models have been developed in due course. In this paper we review the field studies and current knowledge on toxin producing cyanobacterial species and ecological variables that regulate toxin production and bloom formation in major rivers (Han, Geum, Nakdong, and Yeongsan) and reservoirs in South Korea. In addition, development of new, fast, and high-throughput techniques for effective monitoring is also discussed with cyanobacterial bloom advisory practices, current management strategies, and their implications in South Korean freshwater bodies. Ankita Srivastava, Chi-Yong Ahn, Ravi Kumar Asthana, Hyung-Gwan Lee, and Hee-Mock Oh Copyright © 2015 Ankita Srivastava et al. All rights reserved. Salvianolic Acids Attenuate Rat Hippocampal Injury after Acute CO Poisoning by Improving Blood Flow Properties Sun, 01 Feb 2015 09:53:37 +0000 Carbon monoxide (CO) poisoning causes the major injury and death due to poisoning worldwide. The most severe damage via CO poisoning is brain injury and mortality. Delayed encephalopathy after acute CO poisoning (DEACMP) occurs in forty percent of the survivors of acute CO exposure. But the pathological cause for DEACMP is not well understood. And the corresponding therapy is not well developed. In order to investigate the effects of salvianolic acid (SA) on brain injury caused by CO exposure from the view point of hemorheology, we employed a rat model and studied the dynamic of blood changes in the hemorheological and coagulative properties over acute CO exposure. Compared with the groups of CO and 20% mannitol + CO treatments, the severe hippocampal injury caused by acute CO exposure was prevented by SA treatment. These protective effects were associated with the retaining level of hematocrit (Hct), plasma viscosity, fibrinogen, whole blood viscosities and malondialdehyde (MDA) levels in red blood cells (RBCs). These results indicated that SA treatment could significantly improve the deformation of erythrocytes and prevent the damage caused by CO poisoning. Meanwhile, hemorheological indexes are good indicators for monitoring the pathological dynamic after acute CO poisoning. Li Guan, Yan-Lin Zhang, Zong-Yang Li, Ming-Xia Zhu, Wei-Juan Yao, and Jin-Yuan Zhao Copyright © 2015 Li Guan et al. All rights reserved. Transcriptional and Biochemical Effects of Cadmium and Manganese on the Defense System of Octopus vulgaris Paralarvae Thu, 29 Jan 2015 06:59:50 +0000 Due to anthropogenic activities the relative concentrations of cadmium and manganese have increased in the marine environment. Cephalopods are able to accumulate such metals and, as inhabitant of coastal waters, Octopus vulgaris is continuously exposed to anthropogenic activities. Since no study is available on the effects of heavy metals at molecular level in developing octopuses, herein we exposed 1-day-old paralarvae for 24 h to 10, 100, and 1000 μg/L of CdCl2 or MnCl2. Cd exerted a concentration-dependent inhibition of survival and a reduction in growth rate was shown while Mn exposure did not affect the survival rate even at the highest concentrations. Gene expression profiles of hsp70, sod, cat, and gst genes were analyzed by quantitative real-time PCR and defined patterns of transcription were observed. Moreover posttranscriptional analyses were also performed suggesting the impairment of metabolic functions, under strong oxidative conditions (as occurred in paralarvae exposed to Cd) or the complete detoxification events (as occurred in paralarvae exposed to Mn). Aldo Nicosia, Monica Salamone, Salvatore Mazzola, and Angela Cuttitta Copyright © 2015 Aldo Nicosia et al. All rights reserved. Effect of Dietary Lead on Intestinal Nutrient Transporters mRNA Expression in Broiler Chickens Wed, 28 Jan 2015 07:18:23 +0000 Lead- (Pb-) induced oxidative stress is known to suppress growth performance and feed efficiency in broiler chickens. In an attempt to describe the specific underlying mechanisms of such phenomenon we carried out the current study. Ninety-six one-day-old broiler chicks were randomly assigned to 2 dietary treatment groups of 6 pen replicates, namely, (i) basal diet containing no lead supplement (control) and (ii) basal diet containing 200 mg lead acetate/kg of diet. Following 3 weeks of experimental period, jejunum samples were collected to examine the changes in gene expression of several nutrient transporters, antioxidant enzymes, and heat shock protein 70 (Hsp70) using quantitative real-time PCR. The results showed that addition of lead significantly decreased feed intake, body weight gain, and feed efficiency. Moreover, with the exception of GLUT5, the expression of all sugar, peptide, and amino acid transporters was significantly downregulated in the birds under Pb induced oxidative stress. Exposure to Pb also upregulated the antioxidant enzymes gene expression together with the downregulation of glutathione S-transferase and Hsp70. In conclusion, it appears that Pb-induced oxidative stress adversely suppresses feed efficiency and growth performance in chicken and the possible underlying mechanism for such phenomenon is downregulation of major nutrient transporter genes in small intestine. Roohollah Ebrahimi, Mohammad Faseleh Jahromi, Juan Boo Liang, Abdoreza Soleimani Farjam, Parisa Shokryazdan, and Zulkifli Idrus Copyright © 2015 Roohollah Ebrahimi et al. All rights reserved. Effect of Spirulina Intervention on Oxidative Stress, Antioxidant Status, and Lipid Profile in Chronic Obstructive Pulmonary Disease Patients Thu, 22 Jan 2015 13:15:48 +0000 Background and Objective. Oxidative stress is intimately associated with many diseases, including chronic obstructive pulmonary disease (COPD). Study objectives include a comparison of the oxidative stress, antioxidant status, and lipid profile between COPD patients and controls and evaluation of the effect of spirulina intervention on oxidative stress, antioxidant status, and lipid profile of COPD patients. Methods. 30 patients with COPD and 20 controls with no respiratory problems were selected. Global Initiative for Chronic Obstructive Lung Disease criteria were served as the basis of COPD diagnosis. The serum content of malondialdehyde (MDA), lipid hydroperoxide, glutathione (GSH), vitamin C, cholesterol, triglyceride (TG), and high density lipoprotein (HDL) was measured. The activity of superoxide dismutase (SOD), catalase (CAT), and glutathione-s-transferase (GST) was also measured. Two different doses, (500 × 2) mg and (500 × 4) mg spirulina, were given to two groups, each of which comprises 15 COPD patients. Results. All targeted blood parameters have significant difference between COPD patients and controls except triglyceride (TG). Spirulina intake for 30 and 60 days at (500 × 2) mg dose has significantly reduced serum content of MDA, lipid hydroperoxide, and cholesterol while increasing GSH, Vit C level , and the activity of SOD and GST . At the same time, spirulina intake for 30 and 60 days at (500 × 4) mg dose has favorable significant effect on all targeted blood parameters except for HDL . Md. Ismail, Md. Faruk Hossain, Arifur Rahman Tanu, and Hossain Uddin Shekhar Copyright © 2015 Md. Ismail et al. All rights reserved. In Vitro Gender-Dependent Inhibition of Porcine Cytochrome P450 Activity by Selected Flavonoids and Phenolic Acids Wed, 21 Jan 2015 14:20:35 +0000 We investigated gender-related differences in the ability of selected flavonoids and phenolic compounds to modify porcine hepatic CYP450-dependent activity. Using pools of microsomes from male and female pigs, the inhibition of the CYP families 1A, 2A, 2E1, and 3A was determined. The specific CYP activities were measured in the presence of the following selected compounds: rutin, myricetin, quercetin, isorhamnetin, p-coumaric acid, gallic acid, and caffeic acid. We determined that myricetin and isorhamnetin competitively inhibited porcine CYP1A activity in the microsomes from both male and female pigs but did not affect the CYP2A and CYP2E1. Additionally, isorhamnetin competitively inhibited CYP3A in both genders. Noncompetitive inhibition of CYP3A activity by myricetin was observed only in the microsomes from male pigs, whereas CYP3A in female pigs was not affected. Quercetin competitively inhibited CYP2E1 and CYP1A activity in the microsomes from male pigs and irreversibly CY3A in female pigs. No effect of quercetin on CYP2E1 was observed in the microsomes from female pigs. Neither phenolic acids nor rutin affected CYP450 activities. Taken together, our results suggest that the flavonoids myricetin, isorhamnetin, and quercetin may affect the activities of porcine CYP1A, CYP3A, and CYP2E1 in a gender-dependent manner. Bo Ekstrand, Martin Krøyer Rasmussen, Felicia Woll, Vladimir Zlabek, and Galia Zamaratskaia Copyright © 2015 Bo Ekstrand et al. All rights reserved. Soil As Levels and Bioaccumulation in Suaeda salsa and Phragmites australis Wetlands of the Yellow River Estuary, China Wed, 21 Jan 2015 12:34:13 +0000 Little information is available on As contamination dynamics in the soil-plant systems of wetlands. Total arsenic (As) in soil and plant samples from Suaeda salsa and Phragmites australis wetlands was measured in the Yellow River Estuary (YRE) in summer and autumn of 2007 to investigate the seasonal changes in As concentrations in different wetlands. The results showed that soil As levels greatly exceeded the global and regional background values. As levels in soil and the roots and stems of both types of plants were much higher in summer than in autumn, whereas leaf As showed higher level in autumn. Soil sulfur was the main factor influencing As levels in Suaeda salsa wetlands, whereas soil porosity was the most important factor for Phragmites australis wetlands. The contamination factor (CF) showed moderately to considerably polluted levels of As in both wetland soils. Plant roots and leaves of Suaeda salsa had higher As concentrations and biological concentration factors (BCFs) than stems, while the leaves and stems of Phragmites australis showed higher As levels and BCFs than roots. Compared to Phragmites australis, Suaeda salsa generally showed higher translocation factor (TF), while TF values for both plant species were higher in summer than in autumn. Junjing Wang, Junhong Bai, Zhaoqin Gao, Qiongqiong Lu, and Qingqing Zhao Copyright © 2015 Junjing Wang et al. All rights reserved. Acute Toxicity Comparison of Single-Walled Carbon Nanotubes in Various Freshwater Organisms Wed, 14 Jan 2015 14:35:06 +0000 While the commercialization of single-walled carbon nanotubes (SWCNTs) is rapidly expanding, the environmental impact of this nanomaterial is not well understood. Therefore, the present study evaluates the acute aquatic toxicity of SWCNTs towards two freshwater microalgae (Raphidocelis subcapitata and Chlorella vulgaris), a microcrustacean (Daphnia magna), and a fish (Oryzias latipes) based on OECD test guidelines (201, 202, and 203). According to the results, the SWCNTs inhibited the growth of the algae R. subcapitata and C. vulgaris with a median effective concentration (EC50) of 29.99 and 30.96 mg/L, respectively, representing “acute category 3” in the Globally Harmonized System (GHS) of classification and labeling of chemicals. Meanwhile, the acute toxicity test using O. latipes and D. magna did not show any mortality/immobilizing effects up to a concentration of 100.00 mg/L SWCNTs, indicating no hazard category in the GHS classification. In conclusion, SWCNTs were found to induce acute ecotoxicity in freshwater microalgae, yet not in D. magna and medaka fish. Eun Kyung Sohn, Young Shin Chung, Seyed Ali Johari, Tae Gyu Kim, Jin Kwon Kim, Ji Hyun Lee, Yong Hwa Lee, Sung Wook Kang, and Il Je Yu Copyright © 2015 Eun Kyung Sohn et al. All rights reserved. Tetrahydrocannabinol Induces Brain Mitochondrial Respiratory Chain Dysfunction and Increases Oxidative Stress: A Potential Mechanism Involved in Cannabis-Related Stroke Wed, 14 Jan 2015 13:32:09 +0000 Cannabis has potential therapeutic use but tetrahydrocannabinol (THC), its main psychoactive component, appears as a risk factor for ischemic stroke in young adults. We therefore evaluate the effects of THC on brain mitochondrial function and oxidative stress, key factors involved in stroke. Maximal oxidative capacities (complexes I, III, and IV activities), (complexes II, III, and IV activities), (complex IV activity), together with mitochondrial coupling (/), were determined in control conditions and after exposure to THC in isolated mitochondria extracted from rat brain, using differential centrifugations. Oxidative stress was also assessed through hydrogen peroxide (H2O2) production, measured with Amplex Red. THC significantly decreased (−71%; ), (−65%; ), and (−3.5%; ). Mitochondrial coupling (/) was also significantly decreased after THC exposure ( versus ; ). Furthermore, THC significantly enhanced H2O2 production by cerebral mitochondria (+171%; ) and mitochondrial free radical leak was increased from to % (). Thus, THC increases oxidative stress and induces cerebral mitochondrial dysfunction. This mechanism may be involved in young cannabis users who develop ischemic stroke since THC might increase patient’s vulnerability to stroke. Valérie Wolff, Anna-Isabel Schlagowski, Olivier Rouyer, Anne-Laure Charles, François Singh, Cyril Auger, Valérie Schini-Kerth, Christian Marescaux, Jean-Sébastien Raul, Joffrey Zoll, and Bernard Geny Copyright © 2015 Valérie Wolff et al. All rights reserved. Cinnabar-Induced Subchronic Renal Injury Is Associated with Increased Apoptosis in Rats Tue, 06 Jan 2015 06:33:03 +0000 The aim of this study was to explore the role of apoptosis in cinnabar-induced renal injury in rats. To test this role, rats were dosed orally with cinnabar (1 g/kg/day) for 8 weeks or 12 weeks, and the control rats were treated with 5% carboxymethylcellulose solution. Levels of urinary mercury (UHg), renal mercury (RHg), serum creatinine (SCr), and urine kidney injury molecule 1 (KIM-1) were assessed, and renal pathology was analyzed. Apoptotic cells were identified and the apoptotic index was calculated. A rat antibody array was used to analyze expression of cytokines associated with apoptosis. Results from these analyses showed that UHg, RHg, and urine KIM-1, but not SCr, levels were significantly increased in cinnabar-treated rats. Renal pathological changes in cinnabar-treated rats included vacuolization of tubular cells, formation of protein casts, infiltration of inflammatory cells, and increase in the number of apoptotic tubular cells. In comparison to the control group, expression of FasL, Fas, TNF-α, TRAIL, activin A, and adiponectin was upregulated in the cinnabar-treated group. Collectively, our results suggest that prolonged use of cinnabar results in kidney damage due to accumulation of mercury and that the underlying mechanism involves apoptosis of tubular cells via a death receptor-mediated pathway. Ying Wang, Dapeng Wang, Jie Wu, Bohan Wang, Xianhui Gao, Liangjun Wang, and Honglin Ma Copyright © 2015 Ying Wang et al. All rights reserved. Biomarkers of Environmental Pollutants Mon, 22 Dec 2014 12:30:15 +0000 Anilava Kaviraj, Erhan Unlu, Abhik Gupta, and Ahmed El Nemr Copyright © 2014 Anilava Kaviraj et al. All rights reserved. Comparison of the Health Implications on the Use of As and Cd Contaminated Water Supply between Urban and Rural Communities Tue, 04 Nov 2014 00:00:00 +0000 A cross-sectional study was carried out to determine the arsenic (As) and cadmium (Cd) concentrations in blood, urine, and drinking water as well as the health implications on 100 residents in an urban and a rural community. Results showed the blood As, urinary Cd, DNA damage, and water As and Cs were significantly () higher in the rural community. Findings showed significant () correlations between blood As and DNA damage with household income, years of residence, and total glasses of daily water consumption among the rural residents. The urinary NAG concentrations, years of residence, milk powder intake (glass/week), and seafood intake (per week) were significantly correlated () with urinary Cd concentrations among respondents. In addition, urinary Cd level significantly influenced the urinary NAG concentrations (). The rural respondents experienced significantly higher lymphocyte DNA damage and blood As influenced by their years of residence and water consumption. The Cd in drinking water also resulted in the rural respondents having significantly higher urinary NAG which had a significant relationship with urinary Cd. H. Zailina, H. Najibah, A. Nadia Aiezzati, S. M. Praveena, and I. Patimah Copyright © 2014 H. Zailina et al. All rights reserved. In Vivo Protection against Strychnine Toxicity in Mice by the Glycine Receptor Agonist Ivermectin Mon, 15 Sep 2014 09:03:04 +0000 The inhibitory glycine receptor, a ligand-gated ion channel that mediates fast synaptic inhibition in mammalian spinal cord and brainstem, is potently and selectively inhibited by the alkaloid strychnine. The anthelminthic and anticonvulsant ivermectin is a strychnine-independent agonist of spinal glycine receptors. Here we show that ivermectin is an effective antidote of strychnine toxicity in vivo and determine time course and extent of ivermectin protection. Mice received doses of 1 mg/kg and 5 mg/kg ivermectin orally or intraperitoneally, followed by an intraperitoneal strychnine challenge (2 mg/kg). Ivermectin, through both routes of application, protected mice against strychnine toxicity. Maximum protection was observed 14 hours after ivermectin administration. Combining intraperitoneal and oral dosage of ivermectin further improved protection, resulting in survival rates of up to 80% of animals and a significant delay of strychnine effects in up to 100% of tested animals. Strychnine action developed within minutes, much faster than ivermectin, which acted on a time scale of hours. The data agree with a two-compartment distribution of ivermectin, with fat deposits acting as storage compartment. The data demonstrate that toxic effects of strychnine in mice can be prevented if a basal level of glycinergic signalling is maintained through receptor activation by ivermectin. Ahmed Maher, Rasha Radwan, and Hans-Georg Breitinger Copyright © 2014 Ahmed Maher et al. All rights reserved. Acute Toxicity Study of Zerumbone-Loaded Nanostructured Lipid Carrier on BALB/c Mice Model Mon, 08 Sep 2014 11:18:21 +0000 Zerumbone- (ZER-) loaded nanostructure lipid carrier (NLC) (ZER-NLC) prepared for its antileukemia effect in vitro was evaluated for its toxicological effects by observing changes in the liver, kidney, spleen, lung, heart, and brain tissues, serum biochemical parameters, total haemogram, and bone marrow stem cells. The acute toxicity study for ZER-NLC was conducted by orally treating BALB/c mice with a single dose with either water, olive oil, ZER, NLC, or ZER-NLC for 14 days. The animals were observed for clinical and behavioral abnormalities, toxicological symptoms, feed consumption, and gross appearance. The liver, kidney, heart, lung, spleen, and brain tissues were assessed histologically. Total haemogram was counted by hemocytometry and microhematocrit reader. Bone marrow examination in terms of cellular morphology was done by Wright staining with bone marrow smear. Furthermore, serum biochemical parameters were determined spectrophotometrically. Grossly all treated mice, their investigated tissues, serum biochemical parameters, total haemogram, and bone marrow were normal. At oral doses of 100 and 200 mg/kg ZER-NLC there was no sign of toxicity or mortality in BALB/c mice. This study suggests that the 50% lethal dose (LD50) of ZER-NLC is higher than 200 mg/kg, thus, safe by oral administration. Heshu Sulaiman Rahman, Abdullah Rasedee, Hemn Hassan Othman, Max Stanley Chartrand, Farideh Namvar, Swee Keong Yeap, Nozlena Abdul Samad, Reena Joys Andas, Nabilah Muhammad Nadzri, Theebaa Anasamy, Kuan Beng Ng, and Chee Wun How Copyright © 2014 Heshu Sulaiman Rahman et al. All rights reserved. Pollution Status of Pakistan: A Retrospective Review on Heavy Metal Contamination of Water, Soil, and Vegetables Wed, 03 Sep 2014 07:17:49 +0000 Trace heavy metals, such as arsenic, cadmium, lead, chromium, nickel, and mercury, are important environmental pollutants, particularly in areas with high anthropogenic pressure. In addition to these metals, copper, manganese, iron, and zinc are also important trace micronutrients. The presence of trace heavy metals in the atmosphere, soil, and water can cause serious problems to all organisms, and the ubiquitous bioavailability of these heavy metal can result in bioaccumulation in the food chain which especially can be highly dangerous to human health. This study reviews the heavy metal contamination in several areas of Pakistan over the past few years, particularly to assess the heavy metal contamination in water (ground water, surface water, and waste water), soil, sediments, particulate matter, and vegetables. The listed contaminations affect the drinking water quality, ecological environment, and food chain. Moreover, the toxicity induced by contaminated water, soil, and vegetables poses serious threat to human health. Amir Waseem, Jahanzaib Arshad, Farhat Iqbal, Ashif Sajjad, Zahid Mehmood, and Ghulam Murtaza Copyright © 2014 Amir Waseem et al. All rights reserved. Effect of Chlorogenic Acid (5-Caffeoylquinic Acid) Isolated from Baccharis oxyodonta on the Structure and Pharmacological Activities of Secretory Phospholipase A2 from Crotalus durissus terrificus Tue, 02 Sep 2014 13:17:50 +0000 The aim of this paper was to investigate the effect of chlorogenic acid (5-caffeoylquinic acid, 5CQA), isolated from Baccharis oxyodonta, on the structure and pharmacological effect of secretory phospholipase A2 (sPLA2) from Crotalus durissus terrificus. All in vitro and in vivo experiments were conducted using a purified sPLA2 compared under the same experimental conditions with sPLA2 : 5CQA. 5CQA induced several discrete modifications in the secondary structure and the hydrophobic characteristics of native sPLA2 that induced slight changes in the α-helical content, increase in the random coil structure, and decrease of fluorescence of native sPLA2. Moreover, 5CQA significantly decreased the enzymatic activity and the oedema and myonecrosis induced by native sPLA2. As the catalytic activity of sPLA2 plays an important role in several of its biological and pharmacological properties, antibacterial activity was used to confirm the decrease in its enzymatic activity by 5CQA, which induced massive bacterial cell destruction. We found that 5CQA specifically abolished the enzymatic activity of sPLA2 and induced discrete protein unfolding that mainly involved the pharmacological site of sPLA2. These results showed the potential application of 5CQA in the snake poisoning treatment and modulation of the pathological effect of inflammation induced by secretory PLA2. Daniela O. Toyama, Marcelo J. P. Ferreira, Paulete Romoff, Oriana A. Fávero, Henrique H. Gaeta, and Marcos H. Toyama Copyright © 2014 Daniela O. Toyama et al. All rights reserved. The Effect of Multiwalled Carbon Nanotubes on Hepatotoxicity of Cd2+ in Accumulated Cadmium-Metallothione in Mice Tue, 02 Sep 2014 12:45:13 +0000 The effects of oxidized multiwalled carbon nanotubes (oMWCNTs) on the behavior and hepatotoxicity of Cd2+ in accumulated cadmium-metallothionein mice were investigated. The results indicated that, after exposure of oMWCNTs to normal mice, oMWCNTs could not induce the liver to produce metallothionein (MT). When exposing Cd-MT mouse to different doses of oMWCNTs oMWCNTs could cause Cd2+ release from the accumulated Cd-MT; subsequently, one part of the free Cd2+ was eliminated with blood circulation; the other part adsorbed by oMWCNTs would remain in the tissues together with oMWCNTs. The results of the activities changes of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TB), and blood urea nitrogen (BUN) in plasma showed that the hepatotoxicity of coexposure was lower than that of single exposure, and the hepatotoxicity and accumulation of oMWCNTs in livers depended strongly on the exposure dosage of oMWCNTs. The histology of liver and kidney tissue also confirmed the previous results. Therefore, the author inferred that MT could be connected with oMWCNTs to reduce their hepatotoxicity, but the detailed mechanism needs to be further studied. Qi Wei, Bi Juanjuan, Tian Longlong, Li Zhan, Liu Peng, and Wu Wangsuo Copyright © 2014 Qi Wei et al. All rights reserved. Aluminium Involvement in Neurotoxicity Wed, 27 Aug 2014 00:00:00 +0000 The aetiology of neurodegenerative diseases (ND) seems to involve susceptibility genes and environmental factors. Toxic metals are considered major environmental pollutants. Following our study of a case of multiple sclerosis (MS) improvement due to removal of aluminium (Al) and other toxic metals, we have examined the possible relationship between Al intoxication and ND. We used the slow intravenous treatment with the chelating agent EDTA (calcium disodium ethylene diamine tetraacetic acid) (chelation test) to remove Al and detected it in the urine collected from the patients for 12 hours. Patients affected by MS represented 85.6% of total ND. Al was present in 44.8% of cases comprehensive of ND and healthy patients. Al levels were significantly higher in ND patients than in healthy subjects. We here show that treatment of patients affected by Al burden with ten EDTA chelation therapies (EDTA intravenous administration once a week) was able to significantly reduce Al intoxication. Alessandro Fulgenzi, Daniele Vietti, and Maria Elena Ferrero Copyright © 2014 Alessandro Fulgenzi et al. All rights reserved. Modulation of Steroidogenic Pathway in Rat Granulosa Cells with Subclinical Cd Exposure and Insulin Resistance: An Impact on Female Fertility Tue, 19 Aug 2014 06:35:37 +0000 Changes in lifestyle lead to insulin resistance (IR) in females ultimately predisposing them towards infertility. In addition, cadmium (Cd), an environmental endocrine disruptor, is reported for detrimental effects on granulosa cells, thus leading to ovarian dysfunction. A combination of these factors, lifestyle and environment, seems to play a role in etiology of idiopathic infertility that accounts for 50% amongst the total infertility cases. To address this issue, we made an attempt to investigate the extent of Cd impact on insulin-resistant (IR) granulosa cells. We exposed adult female Charles Foster rats to dexamethasone and confirmed IR condition by fasting insulin resistance index (FIRI). On treatment of IR rats with Cd, the preliminary studies demonstrated prolonged estrous cyclicity, decrease in serum estradiol concentrations, abnormal histology of ovary, and increased granulosa cell death. Further gene and protein expression studies of steroidogenic acute regulatory (StAR) protein, 17β-hydroxysteroid dehydrogenase (17β-HSD), and cytochrome P450 aromatase (CYP19A1) were performed. Protein expression studies demonstrated significant decrease in treated groups when compared with control. Study revealed that, in spite of the molecular parameters being affected at varied level, overall ovarian physiology is maximally affected in IR and Cd coexposed group, thus mimicking the condition similar to those prevailing in infertile females. Muskaan Belani, Nupur Purohit, Prakash Pillai, Sharad Gupta, and Sarita Gupta Copyright © 2014 Muskaan Belani et al. All rights reserved. The Presence of Biomarker Enzymes of Selected Scleractinian Corals of Palk Bay, Southeast Coast of India Mon, 18 Aug 2014 12:06:57 +0000 The health and existence of coral reefs are in danger by an increasing range of environmental and anthropogenic impacts. The causes of coral reef decline include worldwide climate change, shoreline development, habitat destruction, pollution, sedimentation and overexploitation. These disasters have contributed to an estimated loss of 27% of the reefs. If the current pressure continues unabated, the estimated loss of coral reef will be about 60% by the year 2030. Therefore, the present study was aimed to analyze the enzymes involved in stress induced by coral pathogen and its resistance. We focused on the enzymes involved in melanin synthesis pathway (phenoloxidase (PO) and peroxidases (POD)) and free radical scavenging enzymes (super oxide dismutase (SOD), catalase (CAT)) and glutathione peroxidase (Gpx) in selected scleractinian corals such as Acropora formosa, Echinopora lamellosa, Favia favus, Favites halicora, Porites sp., and Anacropora forbesi. Overall, PO activity of coral was significantly lower than that of zooxanthellae except for Favia favus. Coral colonies with lower PO and POD activities are prone to disease. Maximum antioxidant defensive enzymes were observed in Favia favus followed by Echinopora lamellose. It is concluded that assay of these enzymes can be used as biomarkers for identifying the susceptibility of corals towards coral bleaching induced by pathogen. R. Anithajothi, K. Duraikannu, G. Umagowsalya, and C. M. Ramakritinan Copyright © 2014 R. Anithajothi et al. All rights reserved. Biomedical Implications of Heavy Metals Induced Imbalances in Redox Systems Tue, 12 Aug 2014 08:42:12 +0000 Several workers have extensively worked out the metal induced toxicity and have reported the toxic and carcinogenic effects of metals in human and animals. It is well known that these metals play a crucial role in facilitating normal biological functions of cells as well. One of the major mechanisms associated with heavy metal toxicity has been attributed to generation of reactive oxygen and nitrogen species, which develops imbalance between the prooxidant elements and the antioxidants (reducing elements) in the body. In this process, a shift to the former is termed as oxidative stress. The oxidative stress mediated toxicity of heavy metals involves damage primarily to liver (hepatotoxicity), central nervous system (neurotoxicity), DNA (genotoxicity), and kidney (nephrotoxicity) in animals and humans. Heavy metals are reported to impact signaling cascade and associated factors leading to apoptosis. The present review illustrates an account of the current knowledge about the effects of heavy metals (mainly arsenic, lead, mercury, and cadmium) induced oxidative stress as well as the possible remedies of metal(s) toxicity through natural/synthetic antioxidants, which may render their effects by reducing the concentration of toxic metal(s). This paper primarily concerns the clinicopathological and biomedical implications of heavy metals induced oxidative stress and their toxicity management in mammals. Bechan Sharma, Shweta Singh, and Nikhat J. Siddiqi Copyright © 2014 Bechan Sharma et al. All rights reserved. Physicochemical Properties of Nanomaterials: Implication in Associated Toxic Manifestations Wed, 06 Aug 2014 08:13:49 +0000 Nanotechnology has emerged as one of the leading fields of the science having tremendous application in diverse disciplines. As nanomaterials are increasingly becoming part of everyday consumer products, it is imperative to assess their impact on living organisms and on the environment. Physicochemical characteristics of nanoparticles and engineered nanomaterials including size, shape, chemical composition, physiochemical stability, crystal structure, surface area, surface energy, and surface roughness generally influence the toxic manifestations of these nanomaterials. This compels the research fraternity to evaluate the role of these properties in determining associated toxicity issues. Reckoning with this fact, in this paper, issues pertaining to the physicochemical properties of nanomaterials as it relates to the toxicity of the nanomaterials are discussed. Manzoor Ahmad Gatoo, Sufia Naseem, Mir Yasir Arfat, Ayaz Mahmood Dar, Khusro Qasim, and Swaleha Zubair Copyright © 2014 Manzoor Ahmad Gatoo et al. All rights reserved. Efficacy of Venom from Tentacle of Jellyfish Stomolophus meleagris (Nemopilema nomurai) against the Cotton Bollworm Helicoverpa armigera Mon, 04 Aug 2014 09:28:34 +0000 Efficacy of venom from tentacle of jellyfish Stomolophus meleagris against the cotton bollworm Helicoverpa armigera was determined. Venom from tentacle of jellyfish Stomolophus meleagris could inhibit the growth of Helicoverpa armigera and the weight inhibiting rate of sample NFr-2 was 60.53%. Of the six samples, only NFr-2 had high insecticidal activity against Helicoverpa armigera and the corrected mortality recorded at 7 d was 74.23%. Huahua Yu, Rongfeng Li, Xiangli Dong, Ronge Xing, Song Liu, and Pengcheng Li Copyright © 2014 Huahua Yu et al. All rights reserved. Azadirachta indica Attenuates Cisplatin-Induced Nephrotoxicity and Oxidative Stress Sun, 03 Aug 2014 09:04:27 +0000 We investigated the effects of methanolic leaves extract of Azadirachta indica (MLEN, 500 mg/kg bwt) on cisplatin- (CP-) induced nephrotoxicity and oxidative stress in rats. CP (5 mg/kg bwt) was injected intraperitoneally and MLEN was given by gastric gavage for 5 days before or after CP injection. After 5 days of CP injection, CP-induced injury of the renal tissue was evidenced (i) as histopathological damage of the renal tissue, (ii) as increases in serum uric acid, urea, and creatinine, (iii) as increases in malondialdehyde (MDA) and nitric oxide (NO), (iv) as decreases in the level of glutathione and activities of superoxide dismutase, catalase, glutathione reductase, glutathione-S-transferase, and glutathione peroxidase, and (v) as increase in the expression of nuclear factor kappa B and apoptosis in kidney tissues. However, the oral administration of MLEN to CP-intoxicated rats for 5 days brought back MDA, NO production, and enzymatic and nonenzymatic antioxidants to near normalcy. Moreover, the histological observations evidenced that neem extract effectively rescues the kidney from CP-mediated oxidative damage. Furthermore, PCR results for caspase-3 and caspase-9 and Bax genes showed downregulation in MLEN treated groups. Therefore, Azadirachta indica can be considered a potential candidate for protection of nephrotoxicity induced by cisplatin. Ahmed E. Abdel Moneim, Mohamed S. Othman, and Ahmed M. Aref Copyright © 2014 Ahmed E. Abdel Moneim et al. All rights reserved. Differences in Cytotoxic, Genotoxic, and Inflammatory Response of Bronchial and Alveolar Human Lung Epithelial Cells to Pristine and COOH-Functionalized Multiwalled Carbon Nanotubes Thu, 24 Jul 2014 00:00:00 +0000 Functionalized MWCNTs are used in many commercial and biomedical applications, but their potential health effects are not well defined. We investigated and compared cytotoxic, genotoxic/oxidative, and inflammatory effects of pristine and carboxyl MWCNTs exposing human respiratory (A549 and BEAS-2B) cells to 1–40 μg/mL of CNTs for 24 h. Both MWCNTs induced low viability reduction (by WST1 assay) in A549 cells and only MWCNTs-COOH caused high viability reduction in BEAS-2B cells reaching 28.5% viability at 40 μg/mL. Both CNTs induced membrane damage (by LDH assay) with higher effects in BEAS-2B cells at the highest concentrations reaching 20% cytotoxicity at 40 μg/mL. DNA damage (by Fpg-comet assay) was induced by pristine MWCNTs in A549 cells and by both MWCNTs in BEAS-2B cells reaching for MWCNTs-COOH a tail moment of 22.2 at 40 μg/mL versus 10.2 of unexposed cells. Increases of IL-6 and IL-8 release (by ELISA) were detected in A549 cells exposed to MWCNTs-COOH from 10 μg/mL while IL-8 increased in BEAS-2B cells exposed to pristine MWCNTs at 20 and 40 μg/mL. The results show higher cytogenotoxicity of MWCNTs-COOH in bronchial and of pristine MWCNTs in alveolar cells. Different inflammatory response was also found. The findings suggest the use of in vitro models with different end points and cells to study CNT toxicity. Cinzia Lucia Ursini, Delia Cavallo, Anna Maria Fresegna, Aureliano Ciervo, Raffaele Maiello, Giuliana Buresti, Stefano Casciardi, Stefano Bellucci, and Sergio Iavicoli Copyright © 2014 Cinzia Lucia Ursini et al. All rights reserved. Venomous and Poisonous Australian Animals of Veterinary Importance: A Rich Source of Novel Therapeutics Mon, 21 Jul 2014 07:59:36 +0000 Envenomation and poisoning by terrestrial animals (both vertebrate and invertebrate) are a significant economic problem and health risk for domestic animals in Australia. Australian snakes are some of the most venomous animals in the world and bees, wasps, ants, paralysis ticks, and cane toads are also present as part of the venomous and poisonous fauna. The diagnosis and treatment of envenomation or poisoning in animals is a challenge and can be a traumatic and expensive process for owners. Despite the potency of Australian venoms, there is potential for novel veterinary therapeutics to be modeled on venom toxins, as has been the case with human pharmaceuticals. A comprehensive overview of envenomation and poisoning signs in livestock and companion animals is provided and related to the potential for venom toxins to act as therapeutics. Margaret C. Hardy, Jonathon Cochrane, and Rachel E. Allavena Copyright © 2014 Margaret C. Hardy et al. All rights reserved. Synergistic Effects of Toxic Elements on Heat Shock Proteins Sun, 20 Jul 2014 09:42:15 +0000 Heat shock proteins show remarkable variations in their expression levels under a variety of toxic conditions. A research span expanded over five decades has revealed their molecular characterization, gene regulation, expression patterns, vast similarity in diverse groups, and broad range of functional capabilities. Their functions include protection and tolerance against cytotoxic conditions through their molecular chaperoning activity, maintaining cytoskeleton stability, and assisting in cell signaling. However, their role as biomarkers for monitoring the environmental risk assessment is controversial due to a number of conflicting, validating, and nonvalidating reports. The current knowledge regarding the interpretation of HSPs expression levels has been discussed in the present review. The candidature of heat shock proteins as biomarkers of toxicity is thus far unreliable due to synergistic effects of toxicants and other environmental factors. The adoption of heat shock proteins as “suit of biomarkers in a set of organisms” requires further investigation. Khalid Mahmood, Saima Jadoon, Qaisar Mahmood, Muhammad Irshad, and Jamshaid Hussain Copyright © 2014 Khalid Mahmood et al. All rights reserved. Chemical Composition, Leishmanicidal and Cytotoxic Activities of the Essential Oils from Mangifera indica L. var. Rosa and Espada Sun, 20 Jul 2014 00:00:00 +0000 The essential oils from Mangifera indica var. Rosa and Espada latex were obtained by hydrodistillation and analyzed using GC-FID and GC-MS. Twenty-seven components were identified. The main compound in the essential oil from M. indica var. Espada (EOMiE) was terpinolene (73.6%). The essential oil of M. indica var. Rosa (EOMiR) was characterized by high amounts of β-pinene (40.7%) and terpinolene (28.3%). In the test for leishmanicidal activity against promastigotes forms of L. amazonensis, EOMiR and EOMiE showed IC50 (72 h) of 39.1 and 23.0 μg/mL, respectively. In macrophages, EOMiR and EOMiE showed CC50 of 142.84 and 158.65 μg/mL, respectively. However, both were more specific to the parasite than macrophages, with values of selectivity index of 6.91 for EOMiE and 3.66 for EOMiR. The essential oils were evaluated for their cytotoxicity against the human tumor cells HEp-2, HT-29, NCI-H292, and HL-60. The EOMiR and EOMiE were most effective against the HL-60, with IC50 values of 12.3 and 3.6 μg/mL, respectively. The results demonstrated that the essential oils of M. indica can destroy L. amazonensis and inhibit tumor cell growth. These findings contribute to the knowledge of the Brazilian biodiversity as a source of potential therapeutic agents. Eduardo H. S. Ramos, Marcílio M. Moraes, Laís L. de A. Nerys, Silene C. Nascimento, Gardênia C. G. Militão, Regina C. B. Q. de Figueiredo, Cláudio A. G. da Câmara, and Teresinha Gonçalves Silva Copyright © 2014 Eduardo H. S. Ramos et al. All rights reserved. Dracocephalum: Novel Anticancer Plant Acting on Liver Cancer Cell Mitochondria Thu, 17 Jul 2014 00:00:00 +0000 Dracocephalum kotschyi Boiss. (Labiatae) is a native Iranian medicinal plant which has been used in combination with Peganum harmala L. as a remedy for many forms of human cancer especially leukemia and gastrointestinal malignancies. Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death worldwide. In this investigation HCC was induced by a single intraperitoneal injection of diethylnitrosamine (DEN) in corn oil at 200 mg/kg body weight to rats. Two weeks after DEN administration, cancer development was promoted with dietary 2-acetylaminofluorene (2-AAF) (0.02%, w/w) for 2 weeks. Serum alpha-fetoprotein (AFP) concentration, serum alanine transaminase (ALT), aspartate transaminase (AST), and alkaline phosphatase (ALP) activities were also determined for confirmation of hepatocellular carcinoma induction. Then rat hepatocytes were isolated with collagen perfusion technique and tumoral hepatocytes were sorted by flow cytometry. Finally isolated mitochondria obtained from both tumoral and nontumoral hepatocytes were used for any probable toxic effect of Dracocephalum kotschyi ethanolic extract. Our results showed that D. kotschyi extract (250 µg/mL) induced reactive oxygen species (ROS) formation, mitochondrial membrane permeabilization (MMP), and mitochondrial swelling and cytochrome c release only in tumoral but not nontumoral hepatocyte. These findings propose Dracocephalum kotschyi as a promising candidate for future anticancer research. Mojtaba Talari, Enayatollah Seydi, Ahmad Salimi, Zhaleh Mohsenifar, Mohammad Kamalinejad, and Jalal Pourahmad Copyright © 2014 Mojtaba Talari et al. All rights reserved. TBT Effects on the Development of Intersex (Ovotestis) in Female Fresh Water Prawn Macrobrachium rosenbergii Thu, 10 Jul 2014 00:00:00 +0000 The impact of tributyltin (TBT) on the female gonad and the endocrine system in Macrobrachium rosenbergii was studied. Prawns were exposed to environmentally realistic concentrations of 10, 100, and 1000 ng/L of TBT for 6 months. Dose dependent effects were noticed in TBT exposed prawns. At 1000 ng/L TBT caused ovotestis formation (formation of male germ cells in ovary). Presence immature oocytes, fusion of developing oocytes, increase in interstitial connective tissues, and its modification into tubular like structure and abundance of spermatogonia in the ovary of TBT treated prawns. The control prawn ovary showed normal architecture of cellular organelles such as mature oocytes with type 2 yolk globules, lipid droplets, normal appearance of yolk envelop, and uniformly arranged microvilli. On the other hand, type 1 yolk globules, reduced size of microvilli, spermatogonial cells in ovary, spermatogonia with centrally located nucleus, and chromatin distribution throughout the nucleoplasm were present in the TBT treated group. Immunofluorescence staining indicated a reduction in vitellin content in ovary of TBT treated prawn. Moreover, TBT had inhibited the vitellogenesis by causing hormonal imbalance in M. rosenbergii. Thus, the present investigation demonstrates that TBT substantially affects sexual differentiation and gonadal development in M. rosenbergii. Revathi Peranandam, Iyapparaj Palanisamy, Arockia Vasanthi Lourdaraj, Munuswamy Natesan, Arun Prasanna Vimalananthan, Suganya Thangaiyan, Anantharaman Perumal, and Krishnan Muthukalingan Copyright © 2014 Revathi Peranandam et al. All rights reserved. Evaluation of Azathioprine-Induced Cytotoxicity in an In Vitro Rat Hepatocyte System Tue, 01 Jul 2014 00:00:00 +0000 Azathioprine (AZA) is widely used in clinical practice for preventing graft rejection in organ transplantations and various autoimmune and dermatological diseases with documented unpredictable hepatotoxicity. The potential molecular cytotoxic mechanisms of AZA towards isolated rat hepatocytes were investigated in this study using “Accelerated Cytotoxicity Mechanism Screening” techniques. The concentration of AZA required to cause 50% cytotoxicity in 2 hrs at 37°C was found to be 400 μM. A significant increase in AZA-induced cytotoxicity and reactive oxygen species (ROS) formation was observed when glutathione- (GSH-) depleted hepatocytes were used. The addition of N-acetylcysteine decreased cytotoxicity and ROS formation. Xanthine oxidase inhibition by allopurinol decreased AZA-induced cytotoxicity, ROS, and hydrogen peroxide (H2O2) formation and increased % mitochondrial membrane potential (MMP). Addition of N-acetylcysteine and allopurinol together caused nearly complete cytoprotection against AZA-induced hepatocyte death. TEMPOL (4-hydroxy-2,2,6,6-tetramethylpiperidin-1-oxyl), a known ROS scavenger and a superoxide dismutase mimic, and antioxidants, like DPPD (N,N′-diphenyl-p-phenylenediamine), Trolox (a water soluble vitamin E analogue), and mesna (2-mercaptoethanesulfonate), also decreased hepatocyte death and ROS formation. Results from this study suggest that AZA-induced cytotoxicity in isolated rat hepatocytes may be partly due to ROS formation and GSH depletion that resulted in oxidative stress and mitochondrial injury. Abdullah Al Maruf, Luke Wan, and Peter J. O’Brien Copyright © 2014 Abdullah Al Maruf et al. All rights reserved. Prophylactic Efficacy of Melatonin on Cyclophosphamide-Induced Liver Toxicity in Mice Mon, 30 Jun 2014 12:10:40 +0000 The current study aimed to evaluate the protective effects of melatonin, a pineal secretory product, against hepatotoxicity induced by cyclophosphamide (CP) in mice. Mice were pretreated with melatonin intraperitoneally for 7 consecutive days before the administration of a single intraperitoneal dose of 200 mg/kg CP. 24 hr after CP administration, the mice were anesthetized, blood was then removed, and serum toxicity enzymes activities were evaluated. After the blood sampling, all animals were killed, livers were then removed, and histological studies were conducted. Serum toxicity marker enzymes were significantly increased after CP treatment but restored in melatonin pretreated groups. In addition, administration of CP induced necrotic hepatocyte with small crushed nuclei, portal space with severe inflammation, and hepatocytes surrounded by lymphocytic infiltration in hepatic tissues. However, melatonin effectively protected against CP-induced histopathological abnormalities in the liver tissues. Our results reveal that melatonin produces a potent hepatoprotective mechanism against CP. Therefore, melatonin could be a potent candidate to use concomitantly as a supplement agent against hepatotoxicity of CP for the patients undergoing chemotherapy. Mohammad Shokrzadeh, Amirhossein Ahmadi, Farshad Naghshvar, Aroona Chabra, and Mehdi Jafarinejhad Copyright © 2014 Mohammad Shokrzadeh et al. All rights reserved. Investigation of Cytotoxicity of Phosphoryl Choline Modified Single-Walled Carbon Nanotubes under a Live Cell Station Wed, 25 Jun 2014 12:10:56 +0000 Single-walled carbon nanotubes (SWCNTs) and various modified SWCNTs have drawn a lot of attention due to their potential applications in biomedical field. Before further moving on to real clinical applications, hydrophobicity and toxicity of SWCNTs should be investigated thoroughly. In this paper, 2-methacryloyloxy ethyl phosphorylcholine (MPC) was adopted to modify SWCNTs and phosphoryl choline was grafted onto SWCNTs as small molecule moieties and polymeric chains, which made SWCNTs dispersed stably both in water and in cell culture medium for a long time. Cytotoxicity of pristine and modified SWCNTs were assayed upon successful preparation of the designed modified SWCNT. Furthermore, the internalization of SWCNTs by three cells was investigated using a live cell station under normal culture temperature (37°C) and low temperature (4°C). The results showed that the internalization of modified SWCNTs was related to both the active transport and the passive transport. Although the modification with phosphoryl choline remarkably reduced the cytotoxicity of SWCNTs, the results were probably due to other reasons such as the decrease in the ratio of cells which internalized modified SWCNTs since the cells without SWCNTs occupation still exhibited normal states. Yufeng Zhao, Qunlong Mao, Yu Liu, Yan Zhang, Tao Zhang, and Zhengsheng Jiang Copyright © 2014 Yufeng Zhao et al. All rights reserved. Neuromodulatory Effects of Hesperidin in Mitigating Oxidative Stress in Streptozotocin Induced Diabetes Thu, 19 Jun 2014 08:27:53 +0000 Oxidative stress has been implicated in pathogenesis of streptozotocin- (STZ-) induced diabetes mellitus and its complication in central nervous system (CNS). Recent studies have provided insights on antioxidants and their emergence as potential therapeutic and nutraceutical. The present study examined the hypothesis that hesperidin (HP) ameliorates oxidative stress and may be a limiting factor in the extent of CNS complication following diabetes. To test this hypothesis rats were divided into four groups: control, diabetic, diabetic-HP treated, and vehicle for HP treatment group. Diabetes mellitus was induced by a single injection of STZ (65 mg/kg body weight). Three days after STZ injection, HP was given (50 mg/kg b.wt. orally) once daily for four weeks. The results of the present investigation suggest that the significant elevated levels of oxidative stress markers were observed in STZ-treated animals, whereas significant depletion in the activity of nonenzymatic antioxidants and enzymatic antioxidants was witnessed in diabetic rat brain. Neurotoxicity biomarker activity was also altered significantly. HP treatment significantly attenuated the altered levels of oxidative stress and neurotoxicity biomarkers. Our results demonstrate that HP exhibits potent antioxidant and neuroprotective effects on the brain tissue against the diabetic oxidative damage in STZ-induced rodent model. Mohammad Ashafaq, Laxmi Varshney, Mohammad Haaris Ajmal Khan, Mohd. Salman, Mehar Naseem, Saima Wajid, and Suhel Parvez Copyright © 2014 Mohammad Ashafaq et al. All rights reserved. Protective Role of Omega-3 Polyunsaturated Fatty Acid against Lead Acetate-Induced Toxicity in Liver and Kidney of Female Rats Wed, 18 Jun 2014 07:13:40 +0000 The present study was conducted to investigate the protective role of Omega-3 polyunsaturated fatty acids against lead acetate-induced toxicity in liver and kidney of female rats. Animals were divided into four equal groups; group 1 served as control while groups 2 and 3 were treated orally with Omega-3 fatty acids at doses of 125 and 260 mg/kg body weight, respectively, for 10 days. These groups were also injected with lead acetate (25 mg/kg body weight) during the last 5 days. Group 4 was treated only with lead acetate for 5 days and served as positive control group. Lead acetate increased oxidative stress through an elevation in MDA associated with depletion in antioxidant enzymes activities in the tissues. Moreover, the elevation of serum enzymes activities (ALT, AST, ALP, and LDH) and the levels of urea and creatinine were estimated but total proteins were decreased. Also, lead acetate-treatment induced hyperlipidemia via increasing of lipid profiles associated with decline in HDL-c level. Significant changes of Hb, PCV, RBCs, PLT, and WBCs in group 4 were recorded. The biochemical alterations of lead acetate were confirmed by histopathological changes and DNA damage. The administration of Omega-3 provided significant protection against lead acetate toxicity. Heba M. Abdou and Mohamed A. Hassan Copyright © 2014 Heba M. Abdou and Mohamed A. Hassan. All rights reserved. Toxic Potential of Synthesized Graphene Zinc Oxide Nanocomposite in the Third Instar Larvae of Transgenic Drosophila melanogaster (hsp70-lacZ)Bg9 Sun, 15 Jun 2014 11:34:38 +0000 In the present study the graphene zinc oxide nanocomposite (GZNC) was synthesized, characterized, and evaluated for its toxic potential on third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ)Bg9. The synthesized GZNC was characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The GZNC in 0.1% dimethyl sulphoxide (DMSO) was sonicated for 10 minutes and the final concentrations 0.033, 0.099, 0.199, and 3.996 μg/μL of diet were established. The third instar larvae were allowed to feed on it separately for 24 and 48 hr. The hsp70 expression was measured by o-nitrophenyl-β-D-galactopyranoside assay, tissue damage was measured by trypan blue exclusion test, and β-galactosidase activity was monitored by in situ histochemical β-galactosidase staining. Oxidative stress was monitored by performing lipid peroxidation assay and total protein estimation. Ethidium bromide/acridine orange staining was performed on midgut cells for apoptotic index and the comet assay was performed for the DNA damage. The results of the present study showed that the exposure of 0.199 and 3.996 μg/μL of GZNC was toxic for both 24 hr and 48 hr of exposure. The doses of 0.033 μg/μL and 0.099 of GZNC showed no toxic effects on its exposure to the third instar larvae for 24 hr as well as 48 hr of duration. Yasir Hasan Siddique, Wasi Khan, Saba Khanam, Smita Jyoti, Falaq Naz, Rahul, Braj Raj Singh, and Alim H. Naqvi Copyright © 2014 Yasir Hasan Siddique et al. All rights reserved. Xanthium strumarium L. Extracts Produce DNA Damage Mediated by Cytotoxicity in In Vitro Assays but Does Not Induce Micronucleus in Mice Sun, 15 Jun 2014 00:00:00 +0000 Xanthium strumarium L. is a member of the Asteraceae commonly used in Cuba, mainly as diuretic. Some toxic properties of this plant have also been reported and, to date, very little is known about its genotoxic properties. The present work aims was to evaluate the potential cytotoxic and genotoxic risk of whole extract from Xanthium strumarium L. whole extract of aerial parts. No positive response was observed in a battery of four Salmonella typhimurium strains, when exposed to concentrations up to 5 mg/plate, with and without mammalian metabolic activation (liver microsomal S9 fraction from Wistar rats). In CHO cells, high concentrations (25–100 μg/mL) revealed significant reduction in cell viability. Results from sister chromatid exchanges, chromosome aberrations, and comet assay showed that X. strumarium extract is genotoxic at the highest concentration used, when clear cytotoxic effects were also observed. On the contrary, no increase in micronuclei frequency in bone marrow cells was observed when the extract was orally administered to mice (100, 500, and 2000 mg/Kg doses). The data presented here constitute the most complete study on the genotoxic potential of X. strumarium L. and show that the extract can induce in vitro DNA damage at cytotoxic concentrations. Janet Piloto Ferrer, Renata Cozzi, Tommaso Cornetta, Pasquale Stano, Mario Fiore, Francesca Degrassi, Rosella De Salvia, Antonia Remigio, Marbelis Francisco, Olga Quiñones, Dayana Valdivia, Maria L. González, Carlos Pérez, and Angel Sánchez-Lamar Copyright © 2014 Janet Piloto Ferrer et al. All rights reserved. Osthole Attenuates Doxorubicin-Induced Apoptosis in PC12 Cells through Inhibition of Mitochondrial Dysfunction and ROS Production Thu, 12 Jun 2014 09:27:44 +0000 Doxorubicin (DOX) is a potent, broad-spectrum chemotherapeutic drug used for treatment of several types of cancers. Despite its effectiveness, it has a wide range of toxic side effects, many of which most likely result from its inherent prooxidant activity. It has been reported that DOX has toxic effects on normal tissues, including brain tissue. In the current study, we investigated the protective effect of osthole isolated from Prangos ferulacea (L.) Lindl. on oxidative stress and apoptosis induced by DOX in PC12 as a neuronal model cell line. PC12 cells were pretreated with osthole 2 h after treatment with different concentrations of DOX. 24 h later, the cell viability, mitochondrial membrane potential (MMP), the activity of caspase-3, the expression ratio of Bax/Bcl-2, and the generation of intracellular ROS were detected. We found that pretreatment with osthole on PC12 cells significantly reduced the loss of cell viability, the activity of caspase-3, the increase in Bax/Bcl-2 ratio, and the generation of intracellular ROS induced by DOX. Moreover, pretreatment with osthole led to an increase in MMP in PC12 cells. In conclusion, our results indicated that pretreatment with nontoxic concentrations of osthole protected PC12 cells from DOX-mediated apoptosis by inhibition of ROS production. Yalda Shokoohinia, Leila Hosseinzadeh, Maryam Moieni-Arya, Ali Mostafaie, and Hamid-Reza Mohammadi-Motlagh Copyright © 2014 Yalda Shokoohinia et al. All rights reserved. Evaluation of Metabolic Enzymes in Response to Excel Mera 71, a Glyphosate-Based Herbicide, and Recovery Pattern in Freshwater Teleostean Fishes Thu, 12 Jun 2014 00:00:00 +0000 Metabolic enzymes such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) were evaluated in Indian teleostean fishes, namely, Anabas testudineus (Bloch) and Heteropneustes fossilis (Bloch), for an exposure to 30 days of Excel Mera 71 (17.2 mg/L), a glyphosate formulation, and subsequent depuration under Liv.52, a plant extract at a dose of 187.5 mg/d/250 L for the same period in the same tissues under laboratory condition. ALT activity was significantly increased () in all the tissues and raised up to 229.19% in liver of A. testudineus (229.19%) and 128.61% in liver of H. fossilis. AST also increased significantly () and was maximum in liver of H. fossilis (526.19%) and minimum in gill of A. testudineus (124.38%). ALP activity was also raised highly in intestine of H. fossilis (490.61%) but was less in kidney of H. fossilis (149.48%). The results indicated that Excel Mera 71 caused alterations in the metabolic enzymatic activities in fish tissues and AST showed the highest alteration in both the fishes, while lowest in ALP and ALT in A. testudineus and H. fossilis, respectively. During depuration under Liv.52, all the enzyme activities came down towards the control condition which indicated the compensatory response by the fish against this herbicidal stress and it was in the following order: AST > ALT > ALP, in A. testudineus, while H. fossilis showed the following trend: ALT > AST > ALP. Therefore, these parameters could be used as indicators of herbicidal pollution in aquatic organisms and were recommended for environmental monitoring for investigating the mechanism involved in the recovery pattern. Palas Samanta, Sandipan Pal, Aloke Kumar Mukherjee, and Apurba Ratan Ghosh Copyright © 2014 Palas Samanta et al. All rights reserved. Toxicity Profile of a Nutraceutical Formulation Derived from Green Mussel Perna viridis Mon, 09 Jun 2014 00:00:00 +0000 The short-term (acute) and long-term (subchronic) toxicity profile, mean lethal dose 50 (LD50), and no-observed-adverse-effect level (NOAEL) of a nutraceutical formulation developed from green mussel Perna viridis, which showed in vitro and in vivo anti-inflammatory properties, were evaluated in the present study. The formulation was administered to the male and female Wistar rats at graded doses (0.5, 1.0, and 2.5 g/kg body weight) for two weeks of acute toxicity study and 0.5, 1.0, and 2.0 g/kg body weight for 90 days in subchronic toxicity study. The LD50, variations in clinical signs, changes in body weight, body weight, food/water consumption, organ weight (liver, kidney, spleen, and brain), hematology, serum chemistry, and histopathological changes were evaluated. The LD50 of the formulation was 5,000 mg/kg BW. No test article related mortalities as well as change in body weight, and food and water consumption were observed. No toxicity related significant changes were noted in renal/hepatic function, hematological indices, and serum biochemical parameters between the control and treated groups. Histopathological alterations were not observed in the vital organs of rats. The subchronic NOAEL for the formulation in rats is greater than 2000 mg/kg. This study demonstrated that the green mussel formulation is safe to consume without any adverse effects in the body. Kajal Chakraborty, Deepu Joseph, and Selsa J. Chakkalakal Copyright © 2014 Kajal Chakraborty et al. All rights reserved. Effect of Resveratrol on Hematological and Biochemical Alterations in Rats Exposed to Fluoride Thu, 05 Jun 2014 12:26:01 +0000 We investigated the protective effects of resveratrol on hematological and biochemical changes induced by fluoride in rats. A total of 28 rats were divided into 4 groups: control, resveratrol, fluoride, and fluoride/resveratrol each), for a total of 21 days of treatment. Blood samples were taken and hematological and biochemical parameters were measured. Compared to the control group, the fluoride-treated group showed significant differences in several hematological parameters, including decreases in WBC, RBC, and PLT counts and neutrophil ratio. The group that received resveratrol alone showed a decrease in WBC count compared to the control group. Furthermore, in comparison to the control group, the fluoride group showed significantly increased ALT enzyme activity and decreased inorganic phosphorus level. The hematological and biochemical parameters in the fluoride + resveratrol treated group were similar to control group. In the fluoride + resveratrol group, resveratrol restored the changes observed following fluoride treatment, including decreased counts of WBC, RBC, and PLT, decreased neutrophil ratio and inorganic phosphorus levels, and elevated ALT enzyme activity. The present study showed that fluoride caused adverse effects in rats and that resveratrol reduced hematological and biochemical alterations produced by fluoride exposure. Nurgül Atmaca, Ebru Yıldırım, Bayram Güner, Ruhi Kabakçı, and Fatih Sultan Bilmen Copyright © 2014 Nurgül Atmaca et al. All rights reserved. Risk Assessment of Heavy Metals Contamination in Paddy Soil, Plants, and Grains (Oryza sativa L.) at the East Coast of India Tue, 03 Jun 2014 11:37:19 +0000 Heavy metals known to be accumulated in plants adversely affect human health. This study aims to assess the effects of agrochemicals especially chemical fertilizers applied in paddy fields, which release potential toxic heavy metals into soil. Those heavy metals get accumulated in different parts of paddy plant (Oryza sativa L.) including the grains. Concentrations of nonessential toxic heavy metals (Cd, Cr, and Pb) and the micronutrients (Cu, Mn, and Zn) were measured in the paddy field soil and plant parts. Mn and Cd are found to be accumulated more in shoot than in root. The metal transfer factors from soil to rice plant were significant for Pb, Cd, Cu, Cr, Mn, and Zn. The ranking order of bioaccumulation factor (BAF) for heavy metals was Zn > Mn > Cd > Cu > Cr > Pb indicating that the accumulation of micronutrients was more than that of nonessential toxic heavy metals. The concentrations of heavy metals were found to be higher in paddy field soils than that of the nearby control soil but below permissible limits. The higher Health Index (HI) values of rice consuming adults (1.561) and children (1.360) suggest their adverse health effects in the near future. Deepmala Satpathy, M. Vikram Reddy, and Soumya Prakash Dhal Copyright © 2014 Deepmala Satpathy et al. All rights reserved. Determination of Mineral, Trace Element, and Pesticide Levels in Honey Samples Originating from Different Regions of Malaysia Compared to Manuka Honey Sun, 01 Jun 2014 11:28:47 +0000 The present study was undertaken to determine the content of six minerals, five trace elements, and ten pesticide residues in honeys originating from different regions of Malaysia. Calcium (Ca), magnesium (Mg), iron (Fe), and zinc (Zn) were analyzed by flame atomic absorption spectrometry (FAAS), while sodium (Na) and potassium (K) were analyzed by flame emission spectrometry (FAES). Trace elements such as arsenic (As), lead (Pb), cadmium (Cd), copper (Cu), and cobalt (Co) were analyzed by graphite furnace atomic absorption spectrometry (GFAAS) following the microwave digestion of honey. High mineral contents were observed in the investigated honeys with K, Na, Ca, and Fe being the most abundant elements (mean concentrations of 1349.34, 236.80, 183.67, and 162.31 mg/kg, resp.). The concentrations of the trace elements were within the recommended limits, indicating that the honeys were of good quality. Principal component analysis reveals good discrimination between the different honey samples. The pesticide analysis for the presence of organophosphorus and carbamates was performed by high performance liquid chromatography (HPLC). No pesticide residues were detected in any of the investigated honey samples, indicating that the honeys were pure. Our study reveals that Malaysian honeys are rich sources of minerals with trace elements present within permissible limits and that they are free from pesticide contamination. Mohammed Moniruzzaman, Muhammed Alamgir Zaman Chowdhury, Mohammad Abdur Rahman, Siti Amrah Sulaiman, and Siew Hua Gan Copyright © 2014 Mohammed Moniruzzaman et al. All rights reserved. Cerium Oxide Nanoparticles Induced Toxicity in Human Lung Cells: Role of ROS Mediated DNA Damage and Apoptosis Sun, 01 Jun 2014 08:12:18 +0000 Cerium oxide nanoparticles (CeO2 NPs) have promising industrial and biomedical applications. In spite of their applications, the toxicity of these NPs in biological/physiological environment is a major concern. Present study aimed to understand the molecular mechanism underlying the toxicity of CeO2 NPs on lung adenocarcinoma (A549) cells. After internalization, CeO2 NPs caused significant cytotoxicity and morphological changes in A549 cells. Further, the cell death was found to be apoptotic as shown by loss in mitochondrial membrane potential and increase in annexin-V positive cells and confirmed by immunoblot analysis of BAX, BCl-2, Cyt C, AIF, caspase-3, and caspase-9. A significant increase in oxidative DNA damage was found which was confirmed by phosphorylation of p53 gene and presence of cleaved poly ADP ribose polymerase (PARP). This damage could be attributed to increased production of reactive oxygen species (ROS) with concomitant decrease in antioxidant “glutathione (GSH)” level. DNA damage and cell death were attenuated by the application of ROS and apoptosis inhibitors N-acetyl-L- cysteine (NAC) and Z-DEVD-fmk, respectively. Our study concludes that ROS mediated DNA damage and cell cycle arrest play a major role in CeO2 NPs induced apoptotic cell death in A549 cells. Apart from beneficial applications, these NPs also impart potential harmful effects which should be properly evaluated prior to their use. Sandeep Mittal and Alok K. Pandey Copyright © 2014 Sandeep Mittal and Alok K. Pandey. All rights reserved. Cytotoxicity of Portuguese Propolis: The Proximity of the In Vitro Doses for Tumor and Normal Cell Lines Sun, 01 Jun 2014 06:36:58 +0000 With a complex chemical composition rich in phenolic compounds, propolis (resinous substance collected by Apis mellifera from various tree buds) exhibits a broad spectrum of biological activities. Recently, in vitro and in vivo data suggest that propolis has anticancer properties, but is the cytoxicity of propolis specific for tumor cells? To answer this question, the cytotoxicity of phenolic extracts from Portuguese propolis of different origins was evaluated using human tumor cell lines (MCF7—breast adenocarcinoma, NCI-H460—non-small cell lung carcinoma, HCT15—colon carcinoma, HeLa—cervical carcinoma, and HepG2—hepatocellular carcinoma), and non-tumor primary cells (PLP2). The studied propolis presented high cytotoxic potential for human tumor cell lines, mostly for HCT15. Nevertheless, excluding HCT15 cell line, the extracts at the GI50 obtained for tumor cell lines showed, in general, cytotoxicity for normal cells (PLP2). Propolis phenolic extracts comprise phytochemicals that should be further studied for their bioactive properties against human colon carcinoma. In the other cases, the proximity of the in vitro cytotoxic doses for tumor and normal cell lines should be confirmed by in vivo tests and may highlight the need for selection of specific compounds within the propolis extract. Ricardo C. Calhelha, Soraia Falcão, Maria João R. P. Queiroz, Miguel Vilas-Boas, and Isabel C. F. R. Ferreira Copyright © 2014 Ricardo C. Calhelha et al. All rights reserved. Evaluating the Effects and Safety of Intravenous Lipid Emulsion on Haloperidol-Induced Neurotoxicity in Rabbit Thu, 29 May 2014 11:58:48 +0000 There are many reports on the effect of intravenous lipid emulsion (ILE) as an antidote in drugs related toxicities. We determined the effects of ILE on neurotoxicity of haloperidol (HA), a highly lipophilic antipsychotic, as a model of antipsychotics poisoning. We used six groups of five male rabbits. Two groups received distilled water intravenously followed by infusions of either 18 mL/kg of normal saline or ILE 20%, after 30 minutes. The third group received 18 mL/kg of normal saline after HA (2.6 mg/kg) administration. The three other groups received ILE 20% solution (6, 12, and 18 mL/kg) following HA injection. Catalepsy scores, temperature, pupil size, and mortality rate were measured at 0, 0.5, 1, 2, 3, 4, 8, and 24 hours after HA administration began. Blood and tissue samples were taken from all animals at 24 hours or at death time for biochemical, cell count, and pathological studies. ILE reversed cataleptic scores, miotic pupils, and hypothermia of HA intoxication much faster than normal saline (). Biochemical complications and mortality rate of the animals were significantly higher in the HA + 18 mL/Kg ILE group. ILE reversed sings of HA neurotoxicity; however, synergistic effect of high dose of ILE and HA increased complications and mortality. Mohammad Moshiri, Amir Hooshang Mohammadpour, Maryam Vahabzadeh, Leila Etemad, Bahram Memar, and Hossein Hosseinzadeh Copyright © 2014 Mohammad Moshiri et al. All rights reserved. Micronucleated Erythrocytes in Newborn Rats Exposed to Raltegravir Placental Transfer Sun, 25 May 2014 08:06:52 +0000 The use of raltegravir in treating HIV/AIDS has been proposed due to its effectiveness in suppressing high loads of HIV RNA in pregnant women, thus preventing infection of the fetus. However, administration of raltegravir during pregnancy produces a compound which is transferred to high concentrations to the offspring. The objective of this study is to evaluate the transplacental genotoxic effect of raltegravir in newborn rats. We evaluated the number of micronucleated erythrocytes (MNE), micronucleated polychromatic erythrocytes (MNPCE), and polychromatic erythrocytes (PCE) in the peripheral blood samples of the offspring of Wistar rats treated 6 days before birth with oral administration of raltegravir. The animals were randomly assigned to five groups as follows: raltegravir at doses of 15, 30, or 60 mg/day, cyclophosphamide 10 mg/kg (positive control), or 0.5 ml of sterile water (negative control). In addition, the effect of these drugs on the weight and height of newborns was assessed. There were no differences in the number of MNE, MNPCE, and PCE, and a slight decrease in the weight and height was observed in the offspring of the rat mothers treated with raltegravir. Genotoxicity studies are required in pregnant women to determine the risk of using raltegravir to the fetuses. Blanca Miriam Torres-Mendoza, Damharis Elizabeth Coronado-Medina, Belinda Claudia Gómez-Meda, Eduardo Vázquez-Valls, Ana Lourdes Zamora-Perez, María de Lourdes Lemus-Varela, and Guillermo Moisés Zúñiga-González Copyright © 2014 Blanca Miriam Torres-Mendoza et al. All rights reserved. Animal Toxins and Their Advantages in Biotechnology and Pharmacology Thu, 22 May 2014 12:45:52 +0000 S. L. Da Silva, E. G. Rowan, F. Albericio, R. G. Stábeli, L. A. Calderon, and A. M. Soares Copyright © 2014 S. L. Da Silva et al. All rights reserved. Genistein Induces Deleterious Effects during Its Acute Exposure in Swiss Mice Thu, 22 May 2014 07:59:00 +0000 Genistein is a soy derived isoflavone. It has wide variety of therapeutic effects against certain diseases including cancer. Although toxic effects of genistein have been studied, its effect on the gene expression and the reason behind toxicity have not been identified yet. In the present study, genistein was administered to age and body weight matched Swiss mice at the doses of 125, 250, 500 and 1000 mg/kg. The biomarkers of hepatotoxicity in serum, liver histology, oxidative stress parameters in tissue homogenates, and global gene expression were examined. Elevated alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) levels and degenerated liver tissue were observed in 500, and 1000 mg/kg genistein treated groups. Oxidative stress was significant at these doses as considerable increase in lipid peroxidation (LPO) and decrease in total glutathione (GSH) were observed. Gene expression analysis showed 40 differentially expressed genes at twofold change and Differentially expressed genes were corresponding to different biologically relevant pathways including metabolic and oxidative stress pathways. In 500 mg/kg group, Cyp4a14, Sult1e1, Gadd45g, Cidec, Mycs, and so forth genes were upregulated. These results suggested that the higher dose of genistein can produce several undesirable effects by affecting multiple cellular pathways. Prabhat Singh, Sharad Sharma, and Srikanta Kumar Rath Copyright © 2014 Prabhat Singh et al. All rights reserved. Toxicology of Metals and Metalloids Thu, 22 May 2014 05:50:09 +0000 Fernando Barbosa Júnior, Marcelo Farina, Susana Viegas, and Wilma De Grava Kempinas Copyright © 2014 Fernando Barbosa Júnior et al. All rights reserved. The β-SiC Nanowires (~100 nm) Induce Apoptosis via Oxidative Stress in Mouse Osteoblastic Cell Line MC3T3-E1 Wed, 21 May 2014 00:00:00 +0000 Silicon carbide (SiC), a compound of silicon and carbon, with chemical formula SiC, the beta modification (β-SiC), with a zinc blende crystal structure (similar to diamond), is formed at temperature below . β-SiC will be the most suitable ceramic material for the future hard tissue replacement, such as bone and tooth. The in vitro cytotoxicity of β-SiC nanowires was investigated for the first time. Our results indicated that 100 nm long SiC nanowires could significantly induce the apoptosis in MC3T3-E1 cells, compared with 100 μm long SiC nanowires. And 100 nm long SiC nanowires increased oxidative stress in MC3T3-E1 cells, as determined by the concentrations of MDA (as a marker of lipid peroxidation) and 8-OHdG (indicator of oxidative DNA damage). Moreover, transmission electron microscopy (TEM) was performed to evaluate the morphological changes of MC3T3-E1 cells. After treatment with 100 nm long SiC nanowires, the mitochondria were swelled and disintegrated, and the production of ATP and the total oxygen uptake were also decreased significantly. Therefore, β-SiC nanowires may have limitations as medical material. Weili Xie, Qi Xie, Meishan Jin, Xiaoxiao Huang, Xiaodong Zhang, Zhengkai Shao, and Guangwu Wen Copyright © 2014 Weili Xie et al. All rights reserved. A Novel Potentiometric Sensor for Determination of Neurotoxin β-N-Oxalyl-L-α, β-Diaminopropionic Acid Tue, 20 May 2014 12:51:29 +0000 A novel potentiometric sensor based on ionophore (Cd(NH2CH2CH2OCH2CH2OCH2CH2NH2)Ag3(CN)5) for the determination of β-N-oxalyl-L-α, β-diaminopropionic acid (ODAP) is developed. The ODAP-selective membrane sensor demonstrates high sensitivity and short response time. The detection limit of the ODAP-selective membrane sensor is about  mol  and the response time is shorter than 6 s. The linear dynamic range of the ODAP-selective membrane sensor is between ODAP concentrations of and  mol . The ODAP-selective membrane sensor exhibits good operational stability for at least one week in dry conditions at 4–6°C. It has a reproducible and stable response during continuous work for at least 10 h with a relative standard deviation of 0.28% ( = 18). Omer Isildak, Furkan Saymaz, Ahmet Karadag, Nesrin Okumus Korkmaz, and Azade Attar Copyright © 2014 Omer Isildak et al. All rights reserved. Assessment of Mutagenic Effect of G. acerosa and S. wightii in S. typhimurium (TA 98, TA 100, and TA 1538 strains) and Evaluation of Their Cytotoxic and Genotoxic Effect in Human Mononuclear Cells: A Non-Clinical Study Tue, 20 May 2014 11:45:15 +0000 The marine red algae (Gelidiella acerosa and Sargassum wightii) possessing excellent antioxidant and anticholinesterase activity were subjected to toxicity evaluation for a deeper understanding of other bioprotective properties of seaweeds. Cytotoxic evaluation was done by trypan blue exclusion, and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays using human PBMC (peripheral blood mononuclear cells) and RBC (red blood cells) lysis assay using human erythrocytes. Mutagenicity of the seaweeds was analyzed by Ames salmonella mutagenicity test with the histidine dependent mutant strains TA 98, TA100 and TA 1538. Genotoxic activity was verified in PBMC by comet assay. The results suggest that benzene extract of G. acerosa (BEGA) and dichloromethane extract of S. wightii (DMESW) did not show cytotoxic effect both in PBMC and erythrocytes. Evaluation of mutagenic activity suggests that the seaweeds did not cause any mutagenic effects both in the absence and the presence of S9 microsomal fraction in all the three Salmonella mutant strains. Results of genotoxic study showed that PBMC treated with seaweed extracts (1 mg/mL) exhibit less or no damage to cells, thus proving the non-genotoxic effect of the extract. Since these in vitro non-clinical studies clearly demonstrate the non-toxic nature of the seaweeds, they could be exploited for further characterization, which would result in development of novel and safe therapeutic entities. Arif Nisha Syad and Pandima Devi Kasi Copyright © 2014 Arif Nisha Syad and Pandima Devi Kasi. All rights reserved. Genotoxicity of Microcystin-LR in In Vitro and In Vivo Experimental Models Sun, 18 May 2014 12:48:04 +0000 Microcystin-LR (MCLR) is a cyanobacterial toxin known for its acute hepatotoxicity. Despite being recognized as tumour promoter, its genotoxicity is far from being completely clarified, particularly in organs other than liver. In this work, we used the comet and/or the micronucleus (MN) assays to study the genotoxicity of MCLR in kidney- (Vero-E6) and liver-derived (HepG2) cell lines and in blood cells from MCLR-exposed mice. MCLR treatment (5 and 20 M) caused a significant induction in the MN frequency in both cell lines and, interestingly, a similar positive effect was observed in mouse reticulocytes (37.5 g MCLR/kg, i.p. route). Moreover, the FISH-based analysis of the MN content (HepG2 cells) suggested that MCLR induces both chromosome breaks and loss. On the other hand, the comet assay results were negative in Vero-E6 cells and in mouse leukocytes, with the exception of a transient increase in the level of DNA damage 30 minutes after mice exposure. Overall, the present findings contributed to increase the weight of evidence in favour of MCLR genotoxicity, based on its capacity to induce permanent genetic damage either in vitro or in vivo. Moreover, they suggest a clastogenic and aneugenic mode of action that might underlie a carcinogenic effect. Elsa Dias, Henriqueta Louro, Miguel Pinto, Telma Santos, Susana Antunes, Paulo Pereira, and Maria João Silva Copyright © 2014 Elsa Dias et al. All rights reserved. Association between Arsenic Exposure and Diabetes Mellitus in Cambodia Sun, 18 May 2014 08:30:10 +0000 Whereas studies in Taiwan found associations between arsenic exposure from drinking water and diabetes mellitus (DM), studies in other countries yielded inconsistent results, and diet might be a confounder. We conducted a study in Cambodia, where people have non-Western style diet, to evaluate the association. We measured well water and urine samples and examined skin signs of arsenicosis to assess arsenic exposure and used questionnaires to collect data on potential risk factors. We performed a fingertip blood glucose test followed by measurement of hemoglobin A1c to assess DM. The 43-male and 99-female participants had an average age of 40.4 years. We found that participants with skin signs of arsenicosis had a higher level of arsenic in the drinking water (1101.1 versus 972.2 μg/L, ). Drinking water with arsenic levels above the median (907.25 μg/L) was associated with a nearly twofold increase in the risk of DM (odds ratio [OR] = 1.7, 95% confidence interval [CI]: 0.5–5.8), so was having skin sings of arsenicosis (OR = 1.7, 95% CI: 0.5–5.6). The ORs did not reach statistical significance most likely because of the small case number. Therefore, further studies with larger study populations are needed to confirm our findings. Jhih-Wei Huang, Ya-Yun Cheng, Tzu-Ching Sung, How-Ran Guo, and Suthipong Sthiannopkao Copyright © 2014 Jhih-Wei Huang et al. All rights reserved. Oxytocin Improves Follicular Reserve in a Cisplatin-Induced Gonadotoxicity Model in Rats Thu, 15 May 2014 00:00:00 +0000 Cisplatin (CP), an antitumor agent, has been shown to cause ovarian injury and dysfunction in both animal and human studies. The present study was conducted to investigate the protective effect of oxytocin (OT) on CP-induced ovarian toxicity in rats. Twenty-one adult female rats were included in the study. Fourteen rats were administered intraperitoneally CP (2 mg/kg/day) twice a week for 5 weeks. Control group () did not receive any treatment. Following treatment, CP-received rats were randomly divided into two groups and treated with either saline (1 mL/kg/day, ) or OT (160 μg/kg/day, ) for 5 weeks. Then, ovarian toxicity and effects of OT were evaluated by histomorphological and biochemical analysis. Our findings revealed a significant reduction in the number of follicles at each grade in saline-treated group. AMH level was significantly lower in saline group compared to control (). OT treatment significantly attenuated CP toxicity in ovaries and increased AMH levels compared to saline group (). Also, administration of OT lessened lipid peroxidation and prevented glutathione depletion in CP-treated rats (). These results indicated that OT could lessen the CP-induced ovarian damage and improve follicular reserve by preventing oxidative damage. Oytun Erbaş, Levent Akman, Altuğ Yavaşoğlu, Mustafa Cosan Terek, Tülay Akman, and Dilek Taskiran Copyright © 2014 Oytun Erbaş et al. All rights reserved. A Positive Babinski Reflex Predicts Delayed Neuropsychiatric Sequelae in Chinese Patients with Carbon Monoxide Poisoning Thu, 15 May 2014 00:00:00 +0000 As the human population increased in China, the carbon monoxide is a serious environmental toxin in public health. However, predicting the delayed neuropsychiatric sequelae (DNS) of carbon monoxide poisoning (COP) has not been well studied. We investigated the independent predictors of DNS in patients with COP. This study was conducted at four hospitals in China. Data were retrospectively collected from 258 patients with COP between November 1990 and October 2011. DNS was the primary endpoint. A positive Babinski reflex was the independent predictor for DNS: sensitivity = 53.8% (95% confidence interval [CI]: 26.1–79.6), specificity = 88.6% (95% CI: 83.7–92.1), positive predictive value (PPV) = 20.0% (95% CI: 9.1–37.5), and negative predictive value (NPV) = 97.3% (95% CI: 94.0–98.9). The area under the receiver operating characteristic curve = 0.712 (95% CI: 0.544–0.880). A positive Babinski reflex was very memorable, immediately available, and applicable in clinical practice. Even when the sensitivity and PPV of a positive Babinski reflex were unsatisfactory, it had a good specificity and NPV for excluding the risk of DNS. In patients without a positive Babinski reflex, the risk for DNS was only 2.7%. This finding may help physicians make decisions about dispositions for patients with COP. Jian-Fang Zou, Qiming Guo, Hua Shao, Bin Li, Yuxiu Du, Maofeng Liu, Fengling Liu, Lixin Dai, Min-Hsien Chung, Hung-Jung Lin, How-Ran Guo, Tzu-Meng Yang, Chien-Cheng Huang, and Chien-Chin Hsu Copyright © 2014 Jian-Fang Zou et al. All rights reserved. Toxicity Evaluation following Intratracheal Instillation of Iron Oxide in a Silica Matrix in Rats Wed, 14 May 2014 10:07:38 +0000 Iron oxide-silica nanoparticles (IOSi-NPs) were prepared from a mixture of ferrous chloride tetrahydrate and ferric chloride hexahydrate dropped into a silica xerogel composite. The structure and morphology of the synthesized maghemite nanoparticles into the silica xerogel were analysed by X-ray diffraction measurements, scanning electron microscopy equipped with an energy dispersive X-ray spectrometer, and transmission electron microscopy. The results of the EDAX analysis indicated that the embedded particles were iron oxide nanoparticles. The particle size of IOSi-NPs calculated from the XRD analysis was estimated at around 12.5 nm. The average size deduced from the particle size distribution is 13.7 ± 0.6 nm, which is in good agreement with XRD analysis. The biocompatibility of IOSi-NPs was assessed by cell viability and cytoskeleton analysis. Histopathology analysis was performed after 24 hours and 7 days, respectively, from the intratracheal instillation of a solution containing 0.5, 2.5, or 5 mg/kg IOSi-NPs. The pathological micrographs of lungs derived from rats collected after the intratracheal instillation with a solution containing 0.5 mg/kg and 2.5 mg/kg IOSi-NPs show that the lung has preserved the architecture of the control specimen with no significant differences. However, even at concentrations of 5 mg/kg, the effect of IOSi-NPS on the lungs was markedly reduced at 7 days posttreatment. Alina Mihaela Prodan, Carmen Steluta Ciobanu, Cristina Liana Popa, Simona Liliana Iconaru, and Daniela Predoi Copyright © 2014 Alina Mihaela Prodan et al. All rights reserved. Punicalagin and Ellagic Acid Demonstrate Antimutagenic Activity and Inhibition of Benzo[a]pyrene Induced DNA Adducts Wed, 14 May 2014 09:47:41 +0000 Punicalagin (PC) is an ellagitannin found in the fruit peel of Punica granatum. We have demonstrated antioxidant and antigenotoxic properties of Punica granatum and showed that PC and ellagic acid (EA) are its major constituents. In this study, we demonstrate the antimutagenic potential, inhibition of BP-induced DNA damage, and antiproliferative activity of PC and EA. Incubation of BP with rat liver microsomes, appropriate cofactors, and DNA in the presence of vehicle or PC and EA showed significant inhibition of the resultant DNA adducts, with essentially complete inhibition (97%) at 40 μM by PC and 77% inhibition by EA. Antimutagenicity was tested by Ames test. PC and EA dose-dependently and markedly antagonized the effect of tested mutagens, sodium azide, methyl methanesulfonate, benzo[a]pyrene, and 2-aminoflourine, with maximum inhibition of mutagenicity up to 90 percent. Almost all the doses tested (50–500 μM) exhibited significant antimutagenicity. A profound antiproliferative effect on human lung cancer cells was also shown with PC and EA. Together, our data show that PC and EA are pomegranate bioactives responsible for inhibition of BP-induced DNA adducts and strong antimutagenic, antiproliferative activities. However, these compounds are to be evaluated in suitable animal model to assess their therapeutic efficacy against cancer. Maryam Zahin, Iqbal Ahmad, Ramesh C. Gupta, and Farrukh Aqil Copyright © 2014 Maryam Zahin et al. All rights reserved. Micronuclei in Bone Marrow and Liver in relation to Hepatic Metabolism and Antioxidant Response due to Coexposure to Chloroform, Dichloromethane, and Toluene in the Rat Model Wed, 14 May 2014 07:19:47 +0000 Genotoxicity in cells may occur in different ways, direct interaction, production of electrophilic metabolites, and secondary genotoxicity via oxidative stress. Chloroform, dichloromethane, and toluene are primarily metabolized in liver by CYP2E1, producing reactive electrophilic metabolites, and may also produce oxidative stress via the uncoupled CYP2E1 catalytic cycle. Additionally, GSTT1 also participates in dichloromethane activation. Despite the oxidative metabolism of these compounds and the production of oxidative adducts, their genotoxicity in the bone marrow micronucleus test is unclear. The objective of this work was to analyze whether the oxidative metabolism induced by the coexposure to these compounds would account for increased micronucleus frequency. We used an approach including the analysis of phase I, phase II, and antioxidant enzymes, oxidative stress biomarkers, and micronuclei in bone marrow (MNPCE) and hepatocytes (MNHEP). Rats were administered different doses of an artificial mixture of CLF/DCM/TOL, under two regimes. After one administration MNPCE frequency increased in correlation with induced GSTT1 activity and no oxidative stress occurred. Conversely, after three-day treatments oxidative stress was observed, without genotoxicity. The effects observed indicate that MNPCE by the coexposure to these VOCs could be increased via inducing the activity of metabolism enzymes. Javier Belmont-Díaz, Ana Paulina López-Gordillo, Eunice Molina Garduño, Luis Serrano-García, Elvia Coballase-Urrutia, Noemí Cárdenas-Rodríguez, Omar Arellano-Aguilar, and Regina D. Montero-Montoya Copyright © 2014 Javier Belmont-Díaz et al. All rights reserved. Alkylation of Histidine Residues of Bothrops jararacussu Venom Proteins and Isolated Phospholipases : A Biotechnological Tool to Improve the Production of Antibodies Sun, 11 May 2014 12:35:29 +0000 Crude venom of Bothrops jararacussu and isolated phospholipases A2 (PLA2) of this toxin (BthTX-I and BthTX-II) were chemically modified (alkylation) by p-bromophenacyl bromide (BPB) in order to study antibody production capacity in function of the structure-function relationship of these substances (crude venom and PLA2 native and alkylated). BthTX-II showed enzymatic activity, while BthTX-I did not. Alkylation reduced BthTX-II activity by 50% while this process abolished the catalytic and myotoxic activities of BthTX-I, while reducing its edema-inducing activity by about 50%. Antibody production against the native and alkylated forms of BthTX-I and -II and the cross-reactivity of antibodies to native and alkylated toxins did not show any apparent differences and these observations were reinforced by surface plasmon resonance (SPR) data. Histopathological analysis of mouse gastrocnemius muscle sections after injection of PBS, BthTX-I, BthTX-II, or both myotoxins previously incubated with neutralizing antibody showed inhibition of the toxin-induced myotoxicity. These results reveal that the chemical modification of the phospholipases A2 (PLA2) diminished their toxicity but did not alter their antigenicity. This observation indicates that the modified PLA2 may provide a biotechnological tool to attenuate the toxicity of the crude venom, by improving the production of antibodies and decreasing the local toxic effects of this poisonous substance in animals used to produce antivenom. C. L. S. Guimarães, S. H. Andrião-Escarso, L. S. Moreira-Dill, B. M. A. Carvalho, D. P. Marchi-Salvador, N. A. Santos-Filho, C. A. H. Fernandes, M. R. M. Fontes, J. R. Giglio, B. Barraviera, J. P. Zuliani, C. F. C. Fernandes, L. A. Calderón, R. G. Stábeli, F. Albericio, S. L. da Silva, and A. M. Soares Copyright © 2014 C. L. S. Guimarães et al. All rights reserved. Fish and Crayfish Toxicology Sun, 11 May 2014 10:10:54 +0000 Josef Velíšek, Zdenka Svobodova, Antonín Kouba, and Zhi-Hua Li Copyright © 2014 Josef Velíšek et al. All rights reserved. Biochemical and Functional Characterization of Parawixia bistriata Spider Venom with Potential Proteolytic and Larvicidal Activities Wed, 07 May 2014 12:41:00 +0000 Toxins purified from the venom of spiders have high potential to be studied pharmacologically and biochemically. These biomolecules may have biotechnological and therapeutic applications. This study aimed to evaluate the protein content of Parawixia bistriata venom and functionally characterize its proteins that have potential for biotechnological applications. The crude venom showed no phospholipase, hemorrhagic, or anti-Leishmania activities attesting to low genotoxicity and discrete antifungal activity for C. albicans. However the following activities were observed: anticoagulation, edema, myotoxicity and proteolysis on casein, azo-collagen, and fibrinogen. The chromatographic and electrophoretic profiles of the proteins revealed a predominance of acidic, neutral, and polar proteins, highlighting the presence of proteins with high molecular masses. Five fractions were collected using cation exchange chromatography, with the P4 fraction standing out as that of the highest purity. All fractions showed proteolytic activity. The crude venom and fractions P1, P2, and P3 showed larvicidal effects on A. aegypti. Fraction P4 showed the presence of a possible metalloprotease (60 kDa) that has high proteolytic activity on azo-collagen and was inhibited by EDTA. The results presented in this study demonstrate the presence of proteins in the venom of P. bistriata with potential for biotechnological applications. Gizeli S. Gimenez, Antonio Coutinho-Neto, Anderson M. Kayano, Rodrigo Simões-Silva, Frances Trindade, Alexandre de Almeida e Silva, Silvana Marcussi, Saulo L. da Silva, Carla F. C. Fernandes, Juliana P. Zuliani, Leonardo A. Calderon, Andreimar M. Soares, and Rodrigo G. Stábeli Copyright © 2014 Gizeli S. Gimenez et al. All rights reserved. Assessment of Acute Oral and Dermal Toxicity of 2 Ethyl-Carbamates with Activity against Rhipicephalus microplus in Rats Tue, 06 May 2014 12:40:42 +0000 The acute oral and dermal toxicity of two new ethyl-carbamates (ethyl-4-bromophenyl-carbamate and ethyl-4-chlorophenyl-carbamate) with ixodicide activity was determined in rats. The oral LD50 of each carbamate was 300 to 2000 mg/kg, and the dermal LD50 of each carbamate was >5000 mg/kg. Clinically, the surviving rats that had received oral doses of each carbamate showed decreased weight gain () and had slight nervous system manifestations. These clinical signs were evident from the 300 mg/kg dose and were reversible, whereas the 2000 mg/kg dose caused severe damage and either caused their death or was motive for euthanasia. At necropsy, these rats had dilated stomachs and cecums with diffuse congestion, as well as moderate congestion of the liver. Histologically, the liver showed slight degenerative lesions, binucleated hepatocytes, focal coagulative necrosis, and congestion areas; the severity of the lesions increased with dosage. Furthermore, an slight increase in gamma-glutamyltransferase, lactate dehydrogenase, and creatinine was observed in the plasma. The dermal application of the maximum dose (5000 mg/kg) of each carbamate did not cause clinical manifestations or liver and skin alterations. This finding demonstrates that the carbamates under study have a low oral hazard and low acute dermal toxicity. María Guadalupe Prado-Ochoa, Ricardo Alfonso Gutiérrez-Amezquita, Víctor Hugo Abrego-Reyes, Ana María Velázquez-Sánchez, Marco Antonio Muñoz-Guzmán, Patricia Ramírez-Noguera, Enrique Angeles, and Fernando Alba-Hurtado Copyright © 2014 María Guadalupe Prado-Ochoa et al. All rights reserved. Age Modulates Fe3O4 Nanoparticles Liver Toxicity: Dose-Dependent Decrease in Mitochondrial Respiratory Chain Complexes Activities and Coupling in Middle-Aged as Compared to Young Rats Tue, 06 May 2014 10:01:33 +0000 We examined the effects of iron oxide nanoparticles (IONPs) on mitochondrial respiratory chain complexes activities and mitochondrial coupling in young (3 months) and middle-aged (18 months) rat liver, organ largely involved in body iron detoxification. Isolated liver mitochondria were extracted using differential centrifugations. Maximal oxidative capacities (, complexes I, III, and IV activities), (complexes II, III, and IV activities), and , (complex IV activity), together with mitochondrial coupling () were determined in controls conditions and after exposure to 250, 300, and 350 μg/ml Fe3O4 in young and middle-aged rats. In young liver mitochondria, exposure to IONPs did not alter mitochondrial function. In contrast, IONPs dose-dependently impaired all complexes of the mitochondrial respiratory chain in middle-aged rat liver: (from 30 ± 1.6 to 17.9 ± 1.5; ), (from 33.9 ± 1.7 to 24.3 ± 1.0; ), (from 43.0 ± 1.6 to 26.3 ± 2.2 µmol O2/min/g protein; ) using Fe3O4 350 µg/ml. Mitochondrial coupling also decreased. Interestingly, 350 μg/ml Fe3O4 in the form of Fe3+ solution did not impair liver mitochondrial function in middle-aged rats. Thus, IONPs showed a specific toxicity in middle-aged rats suggesting caution when using it in old age. Yosra Baratli, Anne-Laure Charles, Valérie Wolff, Lotfi Ben Tahar, Leila Smiri, Jamal Bouitbir, Joffrey Zoll, Mohsen Sakly, Cyril Auger, Thomas Vogel, Hafedh Abdelmelek, Olfa Tebourbi, and Bernard Geny Copyright © 2014 Yosra Baratli et al. All rights reserved.