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    - ENVIRONMENTAL STUDIES (680 journals)
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ENVIRONMENTAL STUDIES (680 journals)            First | 1 2 3 4     

Showing 601 - 378 of 378 Journals sorted alphabetically
Science of The Total Environment     Hybrid Journal   (Followers: 17)
Sciences Eaux & Territoires : la Revue du Cemagref     Open Access  
Scientific Journal of Environmental Sciences     Open Access   (Followers: 1)
Sepsis     Hybrid Journal  
Smart Grid and Renewable Energy     Open Access   (Followers: 8)
Social and Environmental Accountability Journal     Hybrid Journal   (Followers: 2)
Soil and Sediment Contamination: An International Journal     Hybrid Journal   (Followers: 2)
Soil and Tillage Research     Hybrid Journal   (Followers: 6)
SourceOCDE Environnement et developpement durable     Full-text available via subscription   (Followers: 1)
SourceOECD Environment & Sustainable Development     Full-text available via subscription  
South Pacific Journal of Natural and Applied Sciences     Hybrid Journal  
Southern Forests : a Journal of Forest Science     Hybrid Journal   (Followers: 6)
Stochastic Environmental Research and Risk Assessment     Hybrid Journal   (Followers: 4)
Strategic Behavior and the Environment     Full-text available via subscription  
Strategic Planning for Energy and the Environment     Hybrid Journal   (Followers: 4)
Studies in Conservation     Hybrid Journal   (Followers: 11)
Studies in Environmental Science     Full-text available via subscription   (Followers: 6)
Sustainability     Open Access   (Followers: 18)
Sustainability in Environment     Open Access  
Sustainability of Water Quality and Ecology     Hybrid Journal   (Followers: 2)
Sustainable Cities and Society     Hybrid Journal   (Followers: 25)
Sustainable Development     Hybrid Journal   (Followers: 16)
Sustainable Development Law & Policy     Open Access   (Followers: 6)
Sustainable Development Strategy and Practise     Open Access  
Sustainable Environment Research     Open Access  
Sustainable Technologies, Systems & Policies     Open Access   (Followers: 9)
TECHNE - Journal of Technology for Architecture and Environment     Open Access   (Followers: 5)
Tecnogestión     Open Access  
Territorio della Ricerca su Insediamenti e Ambiente. Rivista internazionale di cultura urbanistica     Open Access  
The Historic Environment : Policy & Practice     Hybrid Journal   (Followers: 4)
The International Journal on Media Management     Hybrid Journal   (Followers: 5)
Theoretical Ecology     Hybrid Journal   (Followers: 9)
Theoretical Ecology Series     Full-text available via subscription   (Followers: 1)
Toxicologic Pathology     Hybrid Journal   (Followers: 16)
Toxicological & Environmental Chemistry     Hybrid Journal   (Followers: 4)
Toxicological Sciences     Hybrid Journal   (Followers: 11)
Toxicology     Hybrid Journal   (Followers: 16)
Toxicology and Applied Pharmacology     Hybrid Journal   (Followers: 17)
Toxicology and Industrial Health     Hybrid Journal   (Followers: 7)
Toxicology in Vitro     Hybrid Journal   (Followers: 12)
Toxicology Letters     Hybrid Journal   (Followers: 12)
Toxicology Mechanisms and Methods     Hybrid Journal   (Followers: 10)
Toxicon     Hybrid Journal   (Followers: 3)
Toxin Reviews     Hybrid Journal   (Followers: 1)
Trace Metals and other Contaminants in the Environment     Full-text available via subscription   (Followers: 2)
Trace Metals in the Environment     Full-text available via subscription   (Followers: 2)
Transportation Research Part D: Transport and Environment     Hybrid Journal   (Followers: 26)
Transylvanian Review of Systematical and Ecological Research     Open Access  
Trends in Ecology & Evolution     Full-text available via subscription   (Followers: 174)
Trends in Environmental Analytical Chemistry     Hybrid Journal   (Followers: 2)
Trends in Pharmacological Sciences     Full-text available via subscription   (Followers: 26)
Turkish Journal of Engineering and Environmental Sciences     Open Access   (Followers: 1)
UCLA Journal of Environmental Law and Policy     Open Access   (Followers: 5)
UD y la Geomática     Open Access  
Universidad y Ciencia     Open Access   (Followers: 1)
Urban Studies     Hybrid Journal   (Followers: 49)
Veredas do Direito : Direito Ambiental e Desenvolvimento Sustentável     Open Access  
VertigO - la revue électronique en sciences de l’environnement     Open Access   (Followers: 3)
Villanova Environmental Law Journal     Open Access  
Waste Management & Research     Hybrid Journal   (Followers: 10)
Water Environment Research     Full-text available via subscription   (Followers: 37)
Water International     Hybrid Journal   (Followers: 12)
Water, Air, & Soil Pollution     Hybrid Journal   (Followers: 22)
Water, Air, & Soil Pollution : Focus     Hybrid Journal   (Followers: 9)
Waterlines     Full-text available via subscription   (Followers: 2)
Weather and Forecasting     Full-text available via subscription   (Followers: 15)
Weather, Climate, and Society     Full-text available via subscription   (Followers: 9)
Web Ecology     Open Access   (Followers: 6)
Wetlands     Hybrid Journal   (Followers: 25)
Wilderness & Environmental Medicine     Hybrid Journal   (Followers: 3)
Wildlife Australia     Full-text available via subscription   (Followers: 2)
Wiley Interdisciplinary Reviews - Climate Change     Hybrid Journal   (Followers: 17)
Wiley Interdisciplinary Reviews : Energy and Environment     Hybrid Journal   (Followers: 5)
William & Mary Environmental Law and Policy Review     Open Access   (Followers: 2)
World Environment     Open Access   (Followers: 1)
World Journal of Entrepreneurship, Management and Sustainable Development     Hybrid Journal   (Followers: 4)
World Journal of Environmental Engineering     Open Access   (Followers: 2)
World Journal of Environmental Research     Open Access   (Followers: 1)
Worldviews: Global Religions, Culture, and Ecology     Hybrid Journal   (Followers: 8)
Zoology and Ecology     Hybrid Journal   (Followers: 4)
气候与环境研究     Full-text available via subscription   (Followers: 1)

  First | 1 2 3 4     

Journal Cover Journal of Applied Toxicology
  [SJR: 0.996]   [H-I: 61]   [15 followers]  Follow
   Hybrid Journal Hybrid journal (It can contain Open Access articles)
   ISSN (Print) 0260-437X - ISSN (Online) 1099-1263
   Published by John Wiley and Sons Homepage  [1611 journals]
  • Biological factor related to Asian sand dust particles contributes to the
           exacerbation of asthma
    • Authors: Akiko Honda; Takahiro Sawahara, Tomohiro Hayashi, Kenshi Tsuji, Wataru Fukushima, Mizuki Oishi, Gaku Kitamura, Hitomi Kudo, Sho Ito, Seiichi Yoshida, Takamichi Ichinose, Kayo Ueda, Hirohisa Takano
      Abstract: Epidemiologic studies have revealed that Asian sand dust particles (ASDs) can affect respiratory and immune health represented by asthma. Factors responsible for the exacerbation of asthma remain unclear. The fungus Bjerkandera adusta ( and polycyclic aromatic hydrocarbons such as benzo[a]pyrene (BaP) have been identified in ASDs collected from the atmosphere when an ASD event occurred. We investigated the effects of and BaP related to ASDs on respiratory and immune systems. Bone marrow-derived antigen-presenting cells (APCs) and splenocytes from atopic prone NC/Nga mice and human airway epithelial cells were exposed to the or to BaP in the presence and absence of heated-ASDs (H-ASDs). and BaP in both the presence and absence of H-ASDs increased the expression of cell surface molecules on APCs. H-ASDs alone slightly activated APCs. The expressions induced by were higher than those induced by BaP in the presence and absence of H-ASDs. There were no remarkable effects on the activation of splenocytes or the proinflammatory responses in airway epithelial cells. These results suggest that rather than BaP contributes to the exacerbation of asthma regardless of the presence or absence of sand particles, particularly by the activation of the immune system via APCs. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-10-07T00:26:24.356663-05:
      DOI: 10.1002/jat.3395
  • Immunomodulatory effects of metal salts at sub-toxic concentrations
    • Authors: Carmen Steinborn; Christoph Diegel, Manuel Garcia-Käufer, Carsten Gründemann, Roman Huber
      Abstract: Because different metals are used in complementary medicine for the treatment of diseases related to a dysfunction of the immune system, this study aimed at determining the immunomodulatory potential of Pb(NO3)2, AuCl3, Cu(NO3)2, HgCl2, AgNO3, SnCl2, AsCl3 and SbCl3 at sub-toxic concentrations and at assessing possible toxic side effects of low-concentrated metal preparations. The influence of the metal salts on primary human mononuclear cells was analyzed by measuring cell viability using the water-soluble tetrazolium salt assay, apoptosis and necrosis induction by annexin V/propidium iodide staining and proliferation by carboxyfluorescein diacetate succinimidyl ester staining and flow cytometry. Effects on T-cell activation were assessed with CD69 and CD25 expression using flow cytometry whereas CD83, CD86 and CD14 expression was measured to evaluate the influence on dendritic cell maturation. Alterations of interleukin-2 and interferon-γ secretion were detected by enzyme-linked immunosorbent assay and genotoxic effects were analyzed using the comet assay. At sub-toxic concentrations retardation of T-cell proliferation was caused by Pb(NO3)2, AuCl3 and Cu(NO3)2 and inhibitory effects on interleukin-2 secretion were measured after incubation with Pb(NO3)2, AuCl3, Cu(NO3)2, HgCl2 and AsCl3. Cu(NO3)2 had immunosuppressive activity at dosages within the serum reference range for copper. All other metal salts showed effects at dosages above upper serum limits of normal. Therefore, only low-concentrated copper preparations are promising to have immunomodulatory potential. Toxic side effects of metal preparations used in complementary medicine are improbable because upper limits of metals set in the drinking water ordinance are either not exceeded or the duration of their application is limited. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-10-07T00:16:03.207197-05:
      DOI: 10.1002/jat.3390
  • Comparison of the local pulmonary distribution of nanoparticles
           administered intratracheally to rats via gavage needle or microsprayer
           delivery devices
    • Authors: Guihua Zhang; Naohide Shinohara, Yutaka Oshima, Toshio Kobayashi, Nobuya Imatanaka, Kenji Kawaguchi, Masashi Gamo
      Abstract: Intratracheal administration methods are used to conduct toxicological assessments of inhaled nanoparticles (NPs), and gavage needles or microsprayers are common intratracheal delivery devices. The NP suspension is delivered in a liquid state via gavage needle and as a liquid aerosol via microsprayer. The differences in local pulmonary NP distribution (called the microdistribution) arising from the different states of the NP suspension cause differential pulmonary responses; however, this has yet to be investigated. Herein, using microbeam X-ray fluorescence microscopy, we quantitatively evaluated the TiO2 pulmonary microdistribution (per mesh: 100 μm × 100 μm) in lung sections from rats administered an intratracheal dose of TiO2 NPs (6 mg kg−1) via gavage needle or microsprayer. The results revealed that: (i) using a microsprayer appears to reduce the variations in TiO2 content (ng mesh−1) among rats (e.g., coefficients of variation, n = 3, microsprayer vs gavage needle: 13% vs 30%, for the entire lungs); (ii) TiO2 appears to be deposited less in the right middle lobes than in the rest of the lung lobes, irrespective of the chosen intratracheal delivery device; and (iii) similar TiO2 contents (ng mesh−1) and frequencies are deposited in the lung lobes of rats administered TiO2 NPs via gavage needle or microsprayer. This suggests that the physical state of the administered NP suspension does not markedly alter TiO2 pulmonary microdistribution. The results of this investigation are important for the standardization of intratracheal administration methods. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-10-06T23:04:58.348776-05:
      DOI: 10.1002/jat.3386
  • Disposition of intravenously or orally administered silver nanoparticles
           in pregnant rats and the effect on the biochemical profile in urine
    • Authors: Timothy R. Fennell; Ninell P. Mortensen, Sherry R. Black, Rodney W. Snyder, Keith E. Levine, Eric Poitras, James M. Harrington, Christopher J. Wingard, Nathan A. Holland, Wimal Pathmasiri, Susan C. J. Sumner
      Abstract: Few investigations have been conducted on the disposition and fate of silver nanoparticles (AgNP) in pregnancy. The distribution of a single dose of polyvinylpyrrolidone (PVP)-stabilized AgNP was investigated in pregnant rats. Two sizes of AgNP, 20 and 110 nm, and silver acetate (AgAc) were used to investigate the role of AgNP diameter and particle dissolution in tissue distribution, internal dose and persistence. Dams were administered AgNP or AgAc intravenously (i.v.) (1 mg kg−1) or by gavage (p.o.) (10 mg kg−1), or vehicle alone, on gestation day 18 and euthanized at 24 or 48 h post-exposure. The silver concentration in tissues was measured using inductively-coupled plasma mass spectrometry. The distribution of silver in dams was influenced by route of administration and AgNP size. The highest concentration of silver (μg Ag g−1 tissue) at 48 h was found in the spleen for i.v. administered AgNP, and in the lungs for AgAc. At 48 h after p.o. administration of AgNP, the highest concentration was measured in the cecum and large intestine, and for AgAc in the placenta. Silver was detected in placenta and fetuses for all groups. Markers of cardiovascular injury, oxidative stress marker, cytokines and chemokines were not significantly elevated in exposed dams compared to vehicle-dosed control. NMR metabolomics analysis of urine indicated that AgNP and AgAc exposure impact the carbohydrate, and amino acid metabolism. This study demonstrates that silver crosses the placenta and is transferred to the fetus regardless of the form of silver. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-10-03T03:58:13.817031-05:
      DOI: 10.1002/jat.3387
  • Can CuO nanoparticles lead to epigenetic regulation of antioxidant enzyme
    • Authors: Sandesh Chibber; Rishi Shanker
      Abstract: Copper has been used from ancient time in various applications. Scientists have exploited its means of exposure and consequences to living organisms. The peculiar property of nanomaterials that is a high surface to volume ratio has increased the range of application in products. Copper oxide nanoparticles (CuO NPs) are widely used in industrial applications such as semiconductor devices, gas sensor, batteries, solar energy converter, microelectronics, heat transfer fluids and consumer products. In contrast, acute toxicity of CuO NPs has also been reported. Subsequently, human and environmental health may be at a high risk. Their frequent use can also contaminate ecosystems. Therefore, the toxicity of CuO NPs needs to be thoroughly understood. In this review, we have tried to discuss the recent facts and mechanism that have been explored for CuO NPs-induced toxicity at a cellular, in vivo and ecotoxicological level. Accordingly, the main cause for induction of toxicity by CuO NPs is the generation of reactive oxygen species (ROS) followed by the mitochondrial destruction that leads to apoptosis via the intrinsic pathway or under the condition such as hypoxia cell on exposure to CuO NPs may commit to necrosis. Moreover, CuO NPs also result in activation of MAPK pathways, ERKs and JNK/SAPK thus play an important role in the activation of AP-1. Furthermore, CuO NPs also leads to up-regulation of p53 and caspase three genes. Therefore, careful measures are required to explore omic technology to understand the molecular mechanism of the deleterious effects caused by CuO NPs. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-09-30T03:00:29.591128-05:
      DOI: 10.1002/jat.3392
  • Autophagy function and its relationship to pathology, clinical
           applications, drug metabolism and toxicity
    • Authors: Dayton M. Petibone; Waqar Majeed, Daniel A. Casciano
      Abstract: Autophagy is a cellular process that facilitates nutrient turnover and removal of expended macromolecules and organelles to maintain homeostasis. The recycling of cytosolic macromolecules and damaged organelles by autophagosomes occurs through the lysosomal degradation pathway. Autophagy can also be upregulated as a prosurvival pathway in response to stress stimuli such as starvation, hypoxia or cell damage. Over the last two decades, there has been a surge in research revealing the basic molecular mechanisms of autophagy in mammalian cells. A corollary of an advanced understanding of autophagy has been a concurrent expansion of research into understanding autophagic function and dysfunction in pathology. Recent studies have revealed a pivotal role for autophagy in drug toxicity, and for utilizing autophagic components as diagnostic markers and therapeutic targets in treating disease and cancer. In this review, advances in understanding the molecular basis of mammalian autophagy, methods used to induce and evaluate autophagy, and the diverse interactions between autophagy and drug toxicity, disease progression and carcinogenesis are discussed. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-09-29T03:52:13.343122-05:
      DOI: 10.1002/jat.3393
  • Association between perfluorooctanoic acid exposure and degranulation of
           mast cells in allergic inflammation
    • Authors: Jun-Kyoung Lee; Soyoung Lee, Moon-Chang Baek, Byung-Heon Lee, Hyun-Shik Lee, Taeg Kyu Kwon, Pil-Hoon Park, Tae-Yong Shin, Dongwoo Khang, Sang-Hyun Kim
      Abstract: Perfluorooctanoic acid (PFOA) has wide applications, including as a raw material for converted paper and packaging products. With the widespread use of PFOA, concerns regarding its potential environmental and health impacts have increased. In spite of the known hepatotoxicity and genotoxicity of PFOA, correlation with PFOA and allergic inflammation is not well known. In this study, the effect of PFOA on the degranulation of mast cells and mast cell-mediated allergic inflammation in the presence of FcεRI cross-linking was evaluated. In immunoglobulin (Ig) E-stimulated mast cells, PFOA increased the release of histamine and β-hexosaminidase by the up-regulation of intracellular calcium levels. PFOA enhanced gene expression of several pro-inflammatory cytokines, including tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and IL-8 by the activation of nuclear factor (NF)-κB in IgE-stimulated mast cells. Also, PFOA exacerbated allergic symptoms via hypothermia, and an increase of serum histamine, TNF-α, IgE and IgG1 in the ovalbumin-induced systemic anaphylaxis. The present data indicate that PFOA aggravated FcɛRI-mediated mast cell degranulation and allergic symptoms. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-09-29T03:47:18.076078-05:
      DOI: 10.1002/jat.3389
  • Toxicological role of an acyl glucuronide metabolite in diclofenac-induced
           acute liver injury in mice
    • Authors: Shingo Oda; Yuji Shirai, Sho Akai, Akira Nakajima, Koichi Tsuneyama, Tsuyoshi Yokoi
      Abstract: The acyl glucuronide (AG) metabolites of carboxylic acid-containing drugs are potentially chemically reactive and are suggested to be implicated in toxicity, including hepatotoxicity, nephrotoxicity and drug hypersensitivity reactions. However, it remains unknown whether AG formation is related to toxicity in vivo. In this study, we sought to determine whether AG is involved in the pathogenesis of liver injury using a mouse model of diclofenac (DIC)-induced liver injury. Mice that were administered DIC alone exhibited significantly increased plasma alanine aminotransferase levels, whereas mice that were pretreated with the UDP-glucuronosyltransferase inhibitor (−)-borneol (BOR) exhibited suppressed alanine aminotransferase levels at 3 and 6 h after DIC administration although not significant at 12 h. The plasma DIC-AG concentrations were significantly lower in BOR- and DIC-treated mice than in mice treated with DIC alone. The mRNA expression levels of chemokine (C-X-C motif) ligand 1 (CXCL1), CXCL2 and the neutrophil marker CD11b were reduced in the livers of mice that had been pretreated with BOR compared to those that had been administered DIC alone, whereas mRNA expression of the macrophage marker F4/80 was not altered. An immunohistochemical analysis at 12 h samples revealed that the numbers of myeloperoxidase- and lymphocyte antigen 6 complex-positive cells that infiltrated the liver were significantly reduced in BOR- and DIC-treated mice compared to mice that were treated with DIC alone. These results indicate that DIC-AG is partly involved in the pathogenesis of DIC-induced acute liver injury in mice by activating innate immunity and neutrophils. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-09-27T03:52:22.435402-05:
      DOI: 10.1002/jat.3388
  • Natural remedies for non‐steroidal anti‐inflammatory
           drug‐induced toxicity
    • Authors: Jerine Peter Simon; Sabina Evan Prince
      Abstract: The liver is an important organ of the body, which has a vital role in metabolic functions. The non‐steroidal anti‐inflammatory drug (NSAID), diclofenac causes hepato‐renal toxicity and gastric ulcers. NSAIDs are noted to be an agent for the toxicity of body organs. This review has elaborated various scientific perspectives of the toxicity caused by diclofenac and its mechanistic action in affecting the vital organ. This review suggests natural products are better remedies than current clinical drugs against the toxicity caused by NSAIDs. Natural products are known for their minimal side effects, low cost and availability. On the other hand, synthetic drugs pose the danger of adverse effects if used frequently or over a long period. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-09-22T02:20:46.590665-05:
      DOI: 10.1002/jat.3391
  • Toxic effects of 4‐methylthio‐3‐butenyl isothiocyanate (Raphasatin)
           in the rat urinary bladder without genotoxicity
    • Authors: Isamu Suzuki; Young‐Man Cho, Tadashi Hirata, Takeshi Toyoda, Jun‐ichi Akagi, Yasushi Nakamura, Azusa Sasaki, Takako Nakamura, Shigehisa Okamoto, Koji Shirota, Noboru Suetome, Akiyoshi Nishikawa, Kumiko Ogawa
      Abstract: We recently reported that 4‐methylthio‐3‐butenyl isothiocyanate (MTBITC) exerts chemopreventive effects on the rat esophageal carcinogenesis model at a low dose of 80 ppm in a diet. In contrast, some isothiocyanates (ITCs) have been reported to cause toxic effects, promotion activity, and/or carcinogenic potential in the urinary bladder of rats. In the present study, we investigated whether MTBITC had toxic effects in the urinary bladder similar to other ITCs, such as phenethyl ITC (PEITC). First, to examine the early toxicity of MTBITC, rats were fed a diet supplemented with 100, 300 or 1000 ppm MTBITC for 14 days. Treatment with 1000 ppm MTBITC caused increased organ weights and histopathological changes in the urinary bladder, producing lesions similar to those of 1000 ppm PEITC. In contrast, rats treated with 100 or 300 ppm MTBITC showed no signs of toxicity. Additionally, we performed in vivo genotoxicity studies to clarify whether MTBITC may exhibit a carcinogenic potential through a genotoxic mechanism in rats. Rats were treated with MTBITC for 3 days at doses of 10, 30 or 90 mg kg−1 body weight by gavage, and comet assays in the urinary bladder and micronucleus assays in the bone marrow were performed. No genotoxic changes were observed after treatment with MTBITC at all doses. Overall, these results suggested that the effects of MTBITC in the rat urinary bladder are less than those of PEITC, but that MTBITC could have toxic effects through a nongenotoxic mechanism in the urinary bladder of rats at high doses. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-09-15T23:31:54.945759-05:
      DOI: 10.1002/jat.3384
  • Oxidative stress and cytotoxic effects of silver ion in mouse lung
           macrophages J774.1 cells
    • Authors: Ilseob Shim; Kyunghee Choi, Seishiro Hirano
      Abstract: Silver is commonly used as a disinfectant, and chronic exposure to silver may cause argyria, resulting in a gray–blue discoloration of human skin. However, the mechanism for cellular toxicity of silver has not been well explained. We studied the mode of cell death, the ratio of glutathione disulfide/glutathione, induction of metallothionein and activation of mitogen‐activated protein kinases in J774.1 cells together with activation of antioxidant responsive element and nuclear factor‐κB in CHO cells following exposure to silver ion (Ag+) to investigate the mechanism by which Ag+ causes lethal effects. Ag+ increased phosphorylation levels of extracellular signal‐regulated, c‐Jun N‐terminal and p38 mitogen‐activated protein kinases and remarkably increased the ratio of glutathione disulfide/glutathione in both a time‐ and concentration‐dependent manner. Luciferase reporter gene assays revealed that antioxidant responsive element and nuclear factor‐κB were activated following exposure to Ag+. In addition, exposure to Ag+ increased the mRNA and protein levels of metallothionein. We investigated whether or not Ag+ killed J774.1 cells by inducing apoptosis. Ag+ increased the activity of caspase‐3/7 which was abrogated by caspase 3 and pan‐caspase inhibitors. However, these inhibitors did not ameliorate the cytotoxic effects of Ag+, suggesting that Ag+ causes oxidative stress, which leads to necrotic rather than apoptotic cell death in J774.1 cells by decreasing functional sulfhydryl groups including glutathione in the cells. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-09-14T04:55:49.311991-05:
      DOI: 10.1002/jat.3382
  • Biologic activity of cyclic and caged phosphates: a review
    • Authors: Dietrich E. Lorke; Anka Stegmeier‐Petroianu, Georg A. Petroianu
      Abstract: The recognition in the early 1960s by Morifusa Eto that tri‐o‐cresyl phosphate (TOCP) is hydroxylated by the cytochrome P450 system to an intermediate that spontaneously cyclizes to a neurotoxic phosphate (saligenin phosphate ester) ignited the interest in this group of compounds. Only the ortho isomer can cyclize and clinically cause Organo Phosphate Induced Delayed Neurotoxicity (OPIDN); the meta and para isomers of tri‐cresyl phosphate are not neuropathic because they are unable to form stable cyclic saligenin phosphate esters. This review identifies the diverse biological effects associated with various cyclic and caged phosphates and phosphonates and their possible use. Cyclic compounds that inhibit acetylcholine esterase (AChE), such as salithion, can be employed as pesticides. Others are neurotoxic, most probably because of inhibition of neuropathy target esterase (NTE). Cyclic phosphates that inhibit lipases, the cyclipostins, possibly represent promising therapeutic avenues for the treatment of type 2 diabetes mellitus and/or microbial infections; those compounds inhibiting β‐lactamase may prevent bacterial resistance against β‐lactam antibiotics. Naturally occurring cyclic phosphates, such as cyclic AMP, cyclic phosphatidic acid and the ryanodine receptor modulator cyclic adenosine diphosphate ribose, play an important physiological role in signal transduction. Moreover, some cyclic phosphates are GABA‐antagonists, while others are an essential component of Molybdenum‐containing enzymes. Some cyclic phosphates (cyclophosphamide, ifosfamide) are clinically used in tumor therapy, while the coupling of therapeutic agents with other cyclic phosphates (HepDirect® Technology) allows drugs to be targeted to specific organs. Possible clinical applications of these compounds are considered. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-09-09T11:17:34.349526-05:
      DOI: 10.1002/jat.3369
  • Correlation between antibodies to bisphenol A, its target enzyme protein
           disulfide isomerase and antibodies to neuron‐specific antigens
    • Authors: Datis Kharrazian; Aristo Vojdani
      Abstract: Evidence continues to increase linking autoimmunity and other complex diseases to the chemicals commonly found in our environment. Bisphenol A (BPA) is a synthetic monomer used widely in many forms, from food containers to toys, medical products and many others. The potential for BPA to participate as a triggering agent for autoimmune diseases is likely due to its known immunological influences. The goal of this research was to determine if immune reactivity to BPA has any correlation with neurological antibodies. BPA binds to a target enzyme called protein disulfide isomerase (PDI). Myelin basic protein (MBP) and myelin oligodendrocyte glycoprotein (MOG) are neuronal antigens that are target sites for neuroinflammation and neuroautoimmunity. We determined the co‐occurrence of anti‐MBP and anti‐MOG antibodies with antibodies made against BPA bound to human serum albumin in 100 healthy human subjects. Correlation between BPA to PDI, BPA to MOG, BPA to MBP, PDI to MBP and PDI to MOG were all highly statistically significant (P 
      PubDate: 2016-09-09T11:17:13.249387-05:
      DOI: 10.1002/jat.3383
  • Comparative genotoxicity of silver nanoparticles in human liver HepG2 and
           lung epithelial A549 cells
    • Authors: J. Wang; B. Che, L. W. Zhang, G. Dong, Q. Luo, L. Xin
      Abstract: With the rapid expanding of human exposure to silver nanoparticles (AgNPs), genotoxicity screening of nanosilver is necessary to ensure consumer safety. Here, we assessed one key DNA damage responsive pathway activated by GADD45a gene after 24 h of AgNPs exposure in stable luciferase reporter cell systems based on two widely used in vitro cell models, human liver HepG2 and lung epithelial A549 cells. The comet assay and micronucleus test were also conducted to confirm the genetic damage induced by AgNPs. Our results showed that AgNPs produced a strong dose‐dependent increase in transcriptional activation of GADD45a promoter indicated by luciferase activity accompanying by the significant decreases in cell viability. Surprisingly, in HepG2‐luciferase cells, the relative luciferase activity was greater than 4.5× the control level after being treated with 200 μg ml–1 AgNPs. These results were generally in line with the positive and dose‐dependent responses in cytotoxicity, DNA strand breaks indicated by Olive tail moment, tail DNA (%) and tail length, and chromosome damage indicated by induction of micronuclei, nucleoplasmic bridges, and nuclear buds. Additionally, compared with the A549‐luciferase cells, the HepG2‐luciferase cells seemed to be more susceptible to AgNPs as higher levels of genotoxicity were induced. We concluded that our GADD45a promoter‐driven luciferase reporter gene cell system, together with the comet assay and micronucleus test, can be used as valuable tools for rapid screening of genotoxic potential of nanosilver. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-09-07T06:45:50.883692-05:
      DOI: 10.1002/jat.3385
  • Exposure to cyclic volatile methylsiloxanes (cVMS) causes
           anchorage‐independent growth and reduction of BRCA1 in non‐transformed
           human breast epithelial cells
    • Authors: Abdullah Farasani; Philippa D. Darbre
      Abstract: Dermal absorption of components of personal care products (PCPs) may contribute to breast cancer development. Cyclic volatile methylsiloxanes (cVMS) are used widely in the formulation of PCPs, and their presence has been recently detected in human blood. The objectives of this study were to investigate any genotoxic effects after short‐ (1 week) or longer‐term (30 weeks) exposure to hexamethylcyclotrisiloxane (D3), octamethylcyclotetrasiloxane (D4) or decamethylcyclopentasiloxane (D5) in MCF‐10 A and MCF‐10F immortalized non‐transformed human breast epithelial cells. Genotoxic effects were assessed by an ability of cells to grow in suspension culture, from DNA damage measured by comet assays, and from a reduction in levels of DNA repair proteins measured by RT‐PCR and western immunoblotting. Dose‐dependent anchorage‐independent growth in methocel culture was observed after exposure to D3 (10−13 M–10−5 M) and D4/D5 (10−9 M–10−5 M). DNA damage was measured by the comet assay after 1‐h exposure to D3 (10−6 M–10−5 M) and D4 (10−5 M). BRCA1 mRNA and BRCA1 protein levels were reduced after 30‐week exposure to 10−5 M D4 and D5 in both cell lines. Reduced levels of mRNAs for other DNA repair proteins (BRCA2, ATM, ATR, CHK1 and CHK2) were also observed after exposure to 10−5 M D5 in both cell lines, and some reductions after exposure to D3 and D4. If cVMS can not only enable anchorage‐independent growth of non‐transformed breast epithelial cells and damage DNA, but also compromise DNA repair systems, then there is the potential for them to impact on breast carcinogenesis. Further risk assessment now requires information concerning the extent to which cVMS may be present in human breast tissues. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-09-07T06:30:25.910126-05:
      DOI: 10.1002/jat.3378
  • The role of surface chemistry in the cytotoxicity profile of graphene
    • Authors: Waqar Majeed; Shawn Bourdo, Dayton M. Petibone, Viney Saini, Kieng Bao Vang, Zeid A. Nima, Karrer M. Alghazali, Emilie Darrigues, Anindya Ghosh, Fumiya Watanabe, Daniel Casciano, Syed F. Ali, Alexandru S. Biris
      Abstract: Graphene and its derivative, because of their unique physical, electrical and chemical properties, are an important class of nanomaterials being proposed as foundational materials in nanomedicine as well as for a variety of industrial applications. A major limitation for graphene, when used in biomedical applications, is its poor solubility due to its rather hydrophobic nature. Therefore, chemical functionalities are commonly introduced to alter both its surface chemistry and biochemical activity. Here, we show that surface chemistry plays a major role in the toxicological profile of the graphene structures. To demonstrate this, we chemically increased the oxidation level of the pristine graphene and compared the corresponding toxicological effects along with those for the graphene oxide. X‐ray photoelectron spectroscopy revealed that pristine graphene had the lowest amount of surface oxygen, while graphene oxide had the highest at 2.5% and 31%, respectively. Low and high oxygen functionalized graphene samples were found to have 6.6% and 24% surface oxygen, respectively. Our results showed a dose‐dependent trend in the cytotoxicity profile, where pristine graphene was the most cytotoxic, with decreasing toxicity observed with increasing oxygen content. Increased surface oxygen also played a role in nanomaterial dispersion in water or cell culture medium over longer periods. It is likely that higher dispersity might result in graphene entering into cells as individual flakes ~1 nm thick rather than as more cytotoxic aggregates. In conclusion, changes in graphene's surface chemistry resulted in altered solubility and toxicity, suggesting that a generalized toxicity profile would be rather misleading. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-09-04T22:00:26.495518-05:
      DOI: 10.1002/jat.3379
  • Effects of methyl mercury exposure on pancreatic beta cell development and
    • Authors: Lauren Schumacher; Louise C. Abbott
      Abstract: Methyl mercury is an environmental contaminant of worldwide concern. Since the discovery of methyl mercury exposure due to eating contaminated fish as the underlying cause of the Minamata disaster, the scientific community has known about the sensitivity of the developing central nervous system to mercury toxicity. Warnings are given to pregnant women and young children to limit consumption of foods containing methyl mercury to protect the embryonic, fetal and postnatally developing central nervous system. However, evidence also suggests that exposure to methyl mercury or various forms of inorganic mercury may also affect development and function of other organs. Numerous reports indicate a worldwide increase in diabetes, particularly type 2 diabetes. Quite recently, methyl mercury has been shown to have adverse effects on pancreatic beta (β) cell development and function, resulting in insulin resistance and hyperglycemia and may even lead to the development of diabetes. This review discusses possible mechanisms by which methyl mercury exposure may adversely affect pancreatic β cell development and function, and the role that methyl mercury exposure may have in the reported worldwide increase in diabetes, particularly type 2 diabetes. While additional information is needed regarding associations between mercury exposure and specific mechanisms of the pathogenesis of diabetes in the human population, methyl mercury's adverse effects on the body's natural sources of antioxidants suggest that one possible therapeutic strategy could involve supplementation with antioxidants. Thus, it is important that additional investigation be undertaken into the role of methyl mercury exposure and reduced pancreatic β cell function. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-09-04T22:00:17.284753-05:
      DOI: 10.1002/jat.3381
  • Pharmacokinetics of 14C‐ortho‐phenylphenol following
           intravenous administration in pigs
    • Authors: Emma Nixon; James D. Brooks, Patricia A. Routh, Jason T. Chittenden, Ronald E. Baynes
      Abstract: Workers in the USA are exposed to industrial formulations, which may be toxic. These formulations often contain preservatives or biocides such as ortho‐phenylphenol (OPP). There are limited data describing OPP following intravenous administration to assess truly the clearance of this chemical in humans and other species. In vivo experiments were conducted in pigs to determine related pharmacokinetic parameters. 14C‐OPP was administered as an intravenous bolus dose. Blood, feces, urine and tissue samples were collected for analysis by liquid scintillation. Data were analyzed using non‐compartmental and compartmental pharmacokinetic model approaches. These data fitted a three‐compartment model and showed that the half‐life of 14C‐OPP following the intravenous bolus in pigs was 46.26 ± 10.01 h. The kidneys play a crucial role in clearance of 14C‐OPP with a large percentage of the dose being found in the urine (70.3 ± 6.9% dose). Comparisons with other species suggest that 14C‐OPP clearance in pigs (2.48 ml h–1 kg–1) is less than that in humans (18.87 ml h–1 kg–1) and rats (35.51 ml h–1 kg–1). Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-09-04T22:00:09.282996-05:
      DOI: 10.1002/jat.3380
  • Differences in the mechanisms of action of BDE‐47 and its metabolites on
           OVCAR‐3 and MCF‐7 cell apoptosis
    • Authors: Anna Karpeta; Ewa Łucja Gregoraszczuk
      Abstract: Data concerning possible carcinogenic action of polybrominated diphenyl ethers (PBDEs) in hormone‐dependent tissues are limited. Our earlier studies showed that 2,2′,4,4′‐tetrabromodiphenyl ether (BDE‐47) stimulated OVCAR‐3 and MCF‐7 cell proliferation, while its hydroxylated metabolites (5‐OH‐BDE‐47 and 6‐OH‐BDE‐47) increased estrogen receptors protein expression and extracellular signal‐regulated kinase 1/2 and protein kinase Cα phosphorylation in these cell lines. In addition to cell proliferative disorder, a failure in the regulation of apoptosis can also lead to the formation and development of tumors. Therefore, in the present study, we investigated the effect of BDE‐47 and its metabolites (2.5–50 ng ml–1) on the expression of apoptosis regulatory genes and proteins, caspase‐8 and ‐9 activity and DNA fragmentation induced by extracellular signal‐regulated kinase inhibitor (PD098059) and protein kinase Cα inhibitor (Gӧ 6976) in ovarian (OVCAR‐3) and breast (MCF‐7) cancer cells. In OVCAR‐3 cells, BDE‐47 upregulated expression of most of the investigated genes and increased protein expression of tumor necrosis factor (TNF)‐α, TNF receptor 1, caspase‐6, Bcl‐xl and caspase‐8 activity. Whereas in MCF‐7 cells, BDE‐47 resulted in the downregulation of most of the investigated genes, and decreased caspase‐8 and ‐9 activity. In both OVCAR‐3 and MCF‐7 cells, the expression of most of the investigated genes were downregulated by metabolites. Exposure of OVCAR‐3 cells to 5‐OH‐BDE‐47 corresponded with a decrease in the protein expression of caspase‐6, caspase‐9 and Bcl‐xl and treatment with 6‐OH‐BDE‐47 decreased Bcl‐xl and TNF receptor 1 expression in OVCAR‐3 cells and caspase‐9 expression in MCF‐7 cells. Hydroxylated metabolites of BDE‐47 have strong inhibitory effects on apoptosis in ovarian and breast tumor cells and thus should be considered potential carcinogens in hormone‐dependent cancers. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-09-02T10:05:24.975284-05:
      DOI: 10.1002/jat.3375
  • Methylparaben stimulates tumor initiating cells in ER+ breast cancer
    • Authors: M. Angeles Lillo; Cydney Nichols, Chanel Perry, Stephanie Runke, Raisa Krutilina, Tiffany N. Seagroves, Gustavo A. Miranda‐Carboni, Susan A. Krum
      Abstract: A body of epidemiological evidence implicates exposure to endocrine disrupting chemicals (EDCs) with increased susceptibility to breast cancer. To evaluate the physiological effects of a suspected EDC in vivo, we exposed MCF‐7 breast cancer cells and a patient‐derived xenograft (PDX, estrogen receptor positive) to physiological levels of methylparaben (mePB), which is commonly used in personal care products as a preservative. mePB pellets (4.4 μg per day) led to increased tumor size of MCF‐7 xenografts and ER+ PDX tumors. mePB has been thought to be a xenoestrogen; however, in vitro exposure of 10 nM mePB failed to increase MCF‐7 cell proliferation or induction of canonical estrogen‐responsive genes (pS2 and progesterone receptor), in contrast to 17β‐estradiol (E2) treatment. MCF‐7 and PDX‐derived mammospheres exhibited increased size and up‐regulation of canonical stem cell markers ALDH1, NANOG, OCT4 and SOX2 when exposed to mePB; these effects were not observed for MDA‐MB‐231 (ER−) mammospheres. As tumor‐initiating cells (TICs) are also believed to be responsible for chemoresistance, mammospheres were treated with either tamoxifen or the pure anti‐estrogen fulvestrant in the presence of mePB. Blocking the estrogenic response was not sufficient to block NANOG expression in mammospheres, pointing to a non‐classic estrogen response or an ER‐independent mechanism of mePB promotion of mammosphere activity. Overall, these results suggest that mePB increases breast cancer tumor proliferation through enhanced TIC activity, in part via regulation of NANOG, and that mePB may play a direct role in chemoresistance by modulating stem cell activity. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-09-01T02:21:04.824239-05:
      DOI: 10.1002/jat.3374
  • Observations on conducting whole‐cell patch clamping of the hERG cardiac
           K+ channel in pure human serum
    • Authors: Jiesheng Kang; Yongyi Luo, Michelle Searles, David Rampe
      Abstract: Inhibition of the human ether‐a‐go‐go‐related gene (hERG) K+ channel by drugs leads to QT prolongation on the electrocardiogram and can result in serious cardiac arrhythmia. For this reason, screening of drugs on hERG is mandatory during the drug development process. Patch clamp electrophysiology in a defined physiological saline solution (PSS) represents the standard method for assaying drug effects on the channel. To make the assay more translatable to clinical studies, we have conducted whole‐cell patch clamping of hERG using pure human serum as the extracellular medium. Pure human serum had little effect on the hERG channel waveform or the current–voltage relationship when compared to PSS. hERG current recordings were highly stable in serum at room temperature, but prolonged recordings at the physiological temperature required prior heat inactivation of the serum. Compared to PSS, the IC50 values, conducted at room temperature, of the classic hERG blocking drugs cisapride, moxifloxacin, and terfenadine were shifted to the right by an extent predicted by their known plasma protein binding, but we did not detect any differences in IC50s between male and female serum. Total plasma levels of these drugs associated with clinical QT prolongation corresponded to small (
      PubDate: 2016-08-24T01:35:38.572982-05:
      DOI: 10.1002/jat.3377
  • RNA‐sequencing analysis reveals the hepatotoxic mechanism of
           perfluoroalkyl alternatives, HFPO2 and HFPO4, following exposure in mice
    • Authors: Jianshe Wang; Xiaoyang Wang, Nan Sheng, Xiujuan Zhou, Ruina Cui, Hongxia Zhang, Jiayin Dai
      Abstract: The toxicological impact of traditional perfluoroalkyl chemicals has led to the elimination and restriction of these substances. However, many novel perfluoroalkyl alternatives remain unregulated and little is known about their potential effects on environmental and human health. Daily administration of two alternative perfluoroalkyl substances, HFPO2 and HFPO4 (1 mg kg−1 body weight), for 28 days resulted in hepatomegaly and hepatic histopathological injury in mice, particularly in the HFPO4 group. We generated and compared high‐throughput RNA‐sequencing data from hepatic tissues in control and treatment group mice to clarify the mechanism of HFPO2 and HFPO4 hepatotoxicity. We identified 146 (101 upregulated, 45 downregulated) and 1295 (716 upregulated, 579 downregulated) hepatic transcripts that exhibited statistically significant changes (fold change ≥2 or ≤0.5, false discovery rate 
      PubDate: 2016-08-24T01:25:56.405956-05:
      DOI: 10.1002/jat.3376
  • Comparative ovarian microarray analysis of juvenile hormone‐responsive
           genes in water flea Daphnia magna: potential targets for toxicity
    • Authors: Kenji Toyota; Timothy D. Williams, Tomomi Sato, Norihisa Tatarazako, Taisen Iguchi
      Abstract: The freshwater zooplankton Daphnia magna has been extensively employed in chemical toxicity tests such as OECD Test Guidelines 202 and 211. Previously, it has been demonstrated that the treatment of juvenile hormones (JHs) or their analogues to female daphnids can induce male offspring production. Based on this finding, a rapid screening method for detection of chemicals with JH‐activity was recently developed using adult D. magna. This screening system determines whether a chemical has JH‐activity by investigating the male offspring inducibility. Although this is an efficient high‐throughput short‐term screening system, much remains to be discovered about JH‐responsive pathways in the ovary, and whether different JH‐activators act via the same mechanism. JH‐responsive genes in the ovary including developing oocytes are still largely undescribed. Here, we conducted comparative microarray analyses using ovaries from Daphnia magna treated with fenoxycarb (Fx; artificial JH agonist) or methyl farnesoate (MF; a putative innate JH in daphnids) to elucidate responses to JH agonists in the ovary, including developing oocytes, at a JH‐sensitive period for male sex determination. We demonstrate that induction of hemoglobin genes is a well‐conserved response to JH even in the ovary, and a potential adverse effect of JH agonist is suppression of vitellogenin gene expression, that might cause reduction of offspring number. This is the first report demonstrating different transcriptomics profiles from MF and an artificial JH agonist in D. magna ovary, improving understanding the tissue‐specific mode‐of‐action of JH. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-08-24T01:25:51.702113-05:
      DOI: 10.1002/jat.3368
  • Salinity‐dependent toxicity of water‐dispersible, single‐walled
           carbon nanotubes to Japanese medaka embryos
    • Authors: Chisato Kataoka; Kousuke Nakahara, Kaori Shimizu, Shinsuke Kowase, Seiji Nagasaka, Shinsuke Ifuku, Shosaku Kashiwada
      Abstract: To investigate the effects of salinity on the behavior and toxicity of functionalized single‐walled carbon nanotubes (SWCNTs), which are chemical modified nanotube to increase dispersibility, medaka embryos were exposed to non‐functionalized single‐walled carbon nanotubes (N‐SWCNTs), water‐dispersible, cationic, plastic‐polymer‐coated, single‐walled carbon nanotubes (W‐SWCNTs), or hydrophobic polyethylene glycol‐functionalized, single‐walled carbon nanotubes (PEG‐SWCNTs) at different salinities, from freshwater to seawater. As reference nanomaterials, we tested dispersible chitin nanofiber (CNF), chitosan‐chitin nanofiber (CCNF) and chitin nanocrystal (CNC, i.e. shortened CNF). Under freshwater conditions, with exposure to 10 mg l−1 W‐SWCNTs, the yolk sacks of 57.8% of embryos shrank, and the remaining embryos had a reduced heart rate, eye diameter and hatching rate. Larvae had severe defects of the spinal cord, membranous fin and tail formation. These toxic effects increased with increasing salinity. Survival rates declined with increasing salinity and reached 0.0% in seawater. In scanning electron microscope images, W‐SWCNTs, CNF, CCNF and CNC were adsorbed densely over the egg chorion surface; however, because of chitin's biologically harmless properties, only W‐SWCNTs had toxic effects on the medaka eggs. No toxicity was observed from N‐SWCNT and PEG‐SWCNT exposure. We demonstrated that water dispersibility, surface chemistry, biomedical properties and salinity were important factors in assessing the aquatic toxicity of nanomaterials. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-08-18T04:50:32.43129-05:0
      DOI: 10.1002/jat.3373
  • Sex‐specific characterization and evaluation of the Alzheimer's disease
           genetic risk factor sorl1 in zebrafish during aging and in the adult brain
           following a 100 ppb embryonic lead exposure
    • Authors: Jinyoung Lee; Samuel M. Peterson, Jennifer L. Freeman
      Abstract: Developmental lead (Pb) exposure is suggested in laboratory studies to be a trigger for neurodegenerative diseases such as Alzheimer's disease (AD). Sortilin‐related receptor, L (DLR class) A repeats‐containing (SORL1) is a recently identified AD genetic risk factor. SORL1 has limited characterization in vertebrate models in comparison to other AD genetic risk factors. To characterize SORL1 further, protein sequence homology between humans, mice and zebrafish was analyzed and showed conservation of functional repeats and domain orientation. Next, spatial expression of sorl1 in zebrafish larvae was completed and diffuse expression in neural tissue that was not restricted to the brain was observed. Influences of sex and age on quantitative expression of sorl1 in the brain of adult zebrafish were then assessed. Sex‐specific alteration of sorl1 expression transpired during the aging process in females. The zebrafish was then utilized to investigate the impacts of a 100 ppb embryonic Pb exposure on sorl1 expression and other known AD genetic risk factors. Sex‐specific quantitative gene expression analysis was completed with adult zebrafish brain to compare those developmentally exposed to Pb or a control treatment, but no significant difference in sorl1 expression or other AD genetic risk factors was observed. Overall, this study provided characterization of sorl1 with changes in brain expression during aging being female‐specific. This finding is in agreement with females being more prone to the onset of AD, but analysis of additional AD genetic risk factors is needed to facilitate our understanding of the impact of a 100 ppb embryonic Pb exposure. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-08-18T04:45:32.56419-05:0
      DOI: 10.1002/jat.3372
  • Combining web‐based tools for transparent evaluation of data for risk
           assessment: developmental effects of bisphenol A on the mammary gland as a
           case study
    • Authors: Linda Molander; Annika Hanberg, Christina Rudén, Marlene Ågerstrand, Anna Beronius
      Abstract: Different tools have been developed that facilitate systematic and transparent evaluation and handling of toxicity data in the risk assessment process. The present paper sets out to explore the combined use of two web‐based tools for study evaluation and identification of reliable data relevant to health risk assessment. For this purpose, a case study was performed using in vivo toxicity studies investigating low‐dose effects of bisphenol A on mammary gland development. The reliability of the mammary gland studies was evaluated using the Science in Risk Assessment and Policy (SciRAP) criteria for toxicity studies. The Health Assessment Workspace Collaborative (HAWC) was used for characterizing and visualizing the mammary gland data in terms of type of effects investigated and reported, and the distribution of these effects within the dose interval. It was then investigated whether there was any relationship between study reliability and the type of effects reported and/or their distribution in the dose interval. The combination of the SciRAP and HAWC tools allowed for transparent evaluation and visualization of the studies investigating developmental effects of BPA on the mammary gland. The use of these tools showed that there were no apparent differences in the type of effects and their distribution in the dose interval between the five studies assessed as most reliable and the whole data set. Combining the SciRAP and HAWC tools was found to be a useful approach for evaluating in vivo toxicity studies and identifying reliable and sensitive information relevant to regulatory risk assessment of chemicals. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-08-04T02:55:33.536584-05:
      DOI: 10.1002/jat.3363
  • A modified multiparametric assay using HepaRG cells for predicting the
           degree of drug‐induced liver injury risk
    • Authors: Takafumi Tomida; Hayao Okamura, Tsuyoshi Yokoi, Yoshihiro Konno
      Abstract: The approach for predicting the degree of drug‐induced liver injury (DILI) risk was investigated quantitatively in a modified multiparametric assay using HepaRG cells. Thirty‐eight drugs were classified by DILI risk into five categories based on drug labels approved by the Food and Drug Administration (FDA) as follows: withdrawn (WDN), boxed warning (BW), warnings and precautions (WP), adverse reactions (AR), and no match (NM). Also, WP was classified into two categories: high and low concern. Differentiated HepaRG cells were treated with drugs for 24 h. The maximum concentration was set at 100‐fold the therapeutic maximum plasma concentration (Cmax). After treatment with drugs, the cell viability, glutathione content, caspase 3/7 activity, lactate dehydrogenase leakage and albumin secretion were measured. As modified cut‐off values of each parameter, the TC50 (toxic concentration that decreased the response by 50%) and EC200 (effective concentration giving a response equal to 200% of controls) were calculated. In addition, the toxicity score (total sum score of the cytotoxic level of each parameter) was calculated. This modified multiparametric assay showed an 87% sensitivity and 87% specificity for predicting the DILI risk. The toxicity score showed a good predictive performance for WDN, BW and WP (high concern) categories [cut‐off: score ≥ 1; area under a receiver operating characteristic curve (ROC‐AUC): 0.88], and for WDN and BW categories (cut‐off: score ≥ 3; ROC‐AUC: 0.88). This study newly indicated that the degree of DILI risk might be predictable quantitatively by assessing the toxicity score in the modified multiparametric assay using HepaRG cells. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-08-02T06:16:51.114998-05:
      DOI: 10.1002/jat.3371
  • Cytotoxicity and proliferative capacity impairment induced on human brain
           cell cultures after short‐ and long‐term exposure to magnetite
    • Authors: Teresa Coccini; Francesca Caloni, Lenin Javier Ramírez Cando, Uliana De Simone
      Abstract: Since magnetic iron oxide nanoparticles (IONP) as magnetite (Fe3O4NPs) have potential applications in life sciences, industrial fields and biomedical care, the risks for occupational, general population and patients rises correspondingly. Excessive IONP accumulation in central nervous system (CNS) cells can lead to a disruption of normal iron metabolism/homeostasis, which is a characteristic hallmark resembling that of several neurodegenerative disorders. Fe3O4NPs‐ versus Fe3O4 bulk‐induced toxic effects have been assessed in two human CNS cells namely astrocytes (D384) and neurons (SH‐SY5Y) after short‐term exposure (4–24‐48 h) to 1–100 μg ml−1, and long‐term exposure to lower concentrations. Short‐term Fe3O4NPs induced significant concentration‐ and time‐dependent alterations of mitochondrial function in D384 (25–75% cell viability decrease): effects started at 25 μg ml−1 after 4 h, and 1 μg ml−1 after 48 h. SH‐SY5Y were less susceptible: cytotoxicity occurred after 48  h only with 35–45% mortality (10–100 μg ml−1). Accordingly, a more marked intracellular iron accumulation was observed in astrocytes than neurons. Membrane integrity was unaltered in both CNS cell types. Lowering Fe3O4NP concentrations (0.05–10 μg ml−1) and prolonging the exposure time (up to 10 days), D384 toxicity was again observed (colony number decrease at ≥0.05 μg ml−1, morphology alterations and colony size reduction at ≥0.5 μg ml−1). Effects on SH‐SY5Y appeared at the highest concentration only. Fe3O4 bulk was always remarkably toxic toward both cells. In summary, human cultured astrocytes were susceptible to both Fe3O4NP and bulk forms following short‐term and extended exposure to low concentrations, while neurons were more resistant to NPs. Cellular iron overload may trigger adverse responses by releasing iron ions (particularly in astrocytes) thus compromising the normal functions of CNS. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-08-02T06:06:33.450912-05:
      DOI: 10.1002/jat.3367
  • Multivariate models for prediction of human skin sensitization hazard
    • Authors: Judy Strickland; Qingda Zang, Michael Paris, David M. Lehmann, David Allen, Neepa Choksi, Joanna Matheson, Abigail Jacobs, Warren Casey, Nicole Kleinstreuer
      Abstract: One of the Interagency Coordinating Committee on the Validation of Alternative Method's (ICCVAM) top priorities is the development and evaluation of non‐animal approaches to identify potential skin sensitizers. The complexity of biological events necessary to produce skin sensitization suggests that no single alternative method will replace the currently accepted animal tests. ICCVAM is evaluating an integrated approach to testing and assessment based on the adverse outcome pathway for skin sensitization that uses machine learning approaches to predict human skin sensitization hazard. We combined data from three in chemico or in vitro assays – the direct peptide reactivity assay (DPRA), human cell line activation test (h‐CLAT) and KeratinoSens™ assay – six physicochemical properties and an in silico read‐across prediction of skin sensitization hazard into 12 variable groups. The variable groups were evaluated using two machine learning approaches, logistic regression and support vector machine, to predict human skin sensitization hazard. Models were trained on 72 substances and tested on an external set of 24 substances. The six models (three logistic regression and three support vector machine) with the highest accuracy (92%) used: (1) DPRA, h‐CLAT and read‐across; (2) DPRA, h‐CLAT, read‐across and KeratinoSens; or (3) DPRA, h‐CLAT, read‐across, KeratinoSens and log P. The models performed better at predicting human skin sensitization hazard than the murine local lymph node assay (accuracy 88%), any of the alternative methods alone (accuracy 63–79%) or test batteries combining data from the individual methods (accuracy 75%). These results suggest that computational methods are promising tools to identify effectively the potential human skin sensitizers without animal testing. Published 2016. This article has been contributed to by US Government employees and their work is in the public domain in the USA.
      PubDate: 2016-08-02T06:03:58.640092-05:
      DOI: 10.1002/jat.3366
  • Development of an in vivo anti‐androgenic activity detection assay using
           fenitrothion in Japanese medaka (Oryzias latipes)
    • Authors: Yoshifumi Horie; Haruna Watanabe, Hitomi Takanobu, Ayano Yagi, Takahiro Yamagishi, Taisen Iguchi, Norihisa Tatarazako
      Abstract: The effects of endocrine disruptors, including anti‐androgenic chemicals, on aquatic environments have received increased attention in recent years. Currently, the method used to screen chemicals for anti‐androgenic activity is called the androgenized female stickleback screen, and it was established by the Organization of Economic Cooperation and Development in 2011 using the three‐spined stickleback. However, screening chemicals for anti‐androgenic activity has yet to be established using Japanese medaka. Thus, the purpose of this study was to establish a screening method for anti‐androgenic activity utilizing the number of papillary processes in Japanese medaka (Oryzias latipes) as an indicator of the chemical's anti‐androgenic activity. Thus, at 35 days post‐fertilization, medaka were exposed to fenitrothion, an anti‐androgenic compound, for 28 days. In the control group, the formation of papillary processes was observed in XY medaka, but not in XX medaka. However, after fenitrothion exposure, the number of papillary processes was significantly decreased in a dose‐dependent manner in XY medaka; in the 300 μg l−1 concentration group, four of 11 XY medaka showed no papillary processes even if there were no significant effects on total length and wet body weight compared with the control group. Our results indicate that the number of papillary processes in Japanese medaka can be used as an indicator of anti‐androgenic activity and that this model may prove useful as a chemical screening method. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-07-27T07:20:27.803937-05:
      DOI: 10.1002/jat.3365
  • Gender and geographical variability in the exposure pattern and metabolism
           of deoxynivalenol in humans: a review
    • Authors: Liangkai Chen; Miao Yu, Qinghua Wu, Zhao Peng, Di Wang, Kamil Kuča, Ping Yao, Hong Yan, Andreas K. Nüssler, Liegang Liu, Wei Yang
      Abstract: Deoxynivalenol (DON, also known as vomitoxin) is a common mycotoxin found worldwide, especially in contaminated food. DON is toxic to a variety of cells and tissues in humans. Three kinds of conjugated products (DON‐3‐glucuronide, DON‐15‐glucuronide and DON‐7‐glucuronide) can be found as major metabolites in human urine. Females and males show different patterns of exposure levels, and human exposure to DON also shows some geographical differences because of different DON levels in cereal‐based foods, food intake habits and UDP‐glucuronosyltransferase expression. Specifically, the C12, 13‐deepoxy metabolite was found predominantly in French adults but was rarely detected in UK adults. However, a cohort of Spanish individuals demonstrated even lower DON levels than the levels in the UK populations, whereas a very high DON exposure level was detected in South Africa and Linxian, China. Recent publications have further indicated that DON could be detected in the urine of pregnant women from different countries, which suggests that there is a potential risk to both mothers and foetuses. Additionally, phytochemicals have been shown to be less toxic to cells and laboratory animals in research studies and may also be used as food additives for reducing the toxic effects of DON. In this review, we provide global information on DON metabolism, human exposure and gender differences in humans. Also, control strategies for this mycotoxin are discussed. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-07-26T03:56:47.324771-05:
      DOI: 10.1002/jat.3359
  • Effect of 4‐week inhalation exposure to 1‐bromopropane on
           blood pressure in rats
    • Authors: Fen Huang; Sahoko Ichihara, Yuki Yamada, Shameema Banu, Gaku Ichihara
      Abstract: The pathophysiology of hypertension is complex and multifactorial, and includes exposure to various chemical substances. Several recent studies have documented the reproductive and neurological toxicities of 1‐bromopropane (1‐BP). Given that 1‐BP increased reactive oxygen species in the brain of rats, we hypothesized that 1‐BP also has cardiovascular toxicity through increased oxidative stress. To test this hypothesis, male F344 and Wistar Nagoya rats (n = 7–8 per group per test) were exposed to 0 or 1000 ppm of 1‐BP via inhalation for 4 weeks (8 h per day, 7 days per week). The exposure to 1‐BP increased systolic blood pressure. This effect was associated with a significant decrease in the reduced/oxidized glutathione ratio. A significant increase in nitrotyrosine levels, activation of the NADPH oxidase pathway, which was evidenced by upregulation of gp91phox, a NADPH oxidase subunit, and significant decreases in the expressions of antioxidant molecules such as Cu/Zn‐ and Mn‐superoxide dismutase catalase, and nuclear factor erythroid 2‐related factor 2, were observed in the aortas of Wistar Nagoya rats exposed to 1‐BP. Our results indicate that subacute (4‐week) inhalation exposure to 1‐BP increases blood pressure and suggest that this cardiovascular toxic effect is due, at least in part, to increased oxidative stress mediated through activation of the NADPH oxidase pathway. Further study is needed to assess whether NADPH oxidase activation causes the increase in blood pressure in the rats exposed to 1‐BP. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-07-25T00:30:39.872069-05:
      DOI: 10.1002/jat.3364
  • The C. elegans model in toxicity testing
    • Authors: Piper Reid Hunt
      Abstract: Caenorhabditis elegans is a small nematode that can be maintained at low cost and handled using standard in vitro techniques. Unlike toxicity testing using cell cultures, C. elegans toxicity assays provide data from a whole animal with intact and metabolically active digestive, reproductive, endocrine, sensory and neuromuscular systems. Toxicity ranking screens in C. elegans have repeatedly been shown to be as predictive of rat LD50 ranking as mouse LD50 ranking. Additionally, many instances of conservation of mode of toxic action have been noted between C. elegans and mammals. These consistent correlations make the case for inclusion of C. elegans assays in early safety testing and as one component in tiered or integrated toxicity testing strategies, but do not indicate that nematodes alone can replace data from mammals for hazard evaluation. As with cell cultures, good C. elegans culture practice (GCeCP) is essential for reliable results. This article reviews C. elegans use in various toxicity assays, the C. elegans model's strengths and limitations for use in predictive toxicology, and GCeCP. Published 2016. This article is a U.S. Government work and is in the public domain in the USA. Journal of Applied Toxicology published by John Wiley & Sons Ltd.
      PubDate: 2016-07-22T01:45:37.492847-05:
      DOI: 10.1002/jat.3357
  • Pulmonary persistence of graphene nanoplatelets may disturb physiological
           and immunological homeostasis
    • Authors: Eun‐Jung Park; Sang Jin Lee, Kyuhong Lee, Young Chul Choi, Byoung‐Seok Lee, Gwang‐Hee Lee, Dong‐Wan Kim
      Abstract: Accumulated evidence suggests that chronic pulmonary accumulation of harmful particles cause adverse pulmonary and systemic health effects. In our previous study, most of the graphene nanoplatelet (GNP) remained in the lung until 28 days after a single instillation. In this study, we sought to evaluate the local and systemic health effect after a long pulmonary persistence of GNP. As expected, GNP remained in the lung on day 90 after a single intratracheal instillation (1.25, 2.5 and 5 mg kg−1). In the lung exposed at the highest dose, the total number of cells and the percentage of lymphocytes significantly increased in the BAL fluid with an increase in both the number of GNP‐engulfed macrophages and the percentage of apoptotic cells. A Th1‐shifted immune response, the elevated chemokine secretion and the enhanced expression of cytoskeletal‐related genes were observed. Additionally, the expression of natriuretic‐related genes was noteworthy altered in the lungs. Moreover, the number of white blood cells (WBC) and the percentage of macrophages and neutrophils clearly increased in the blood of mice exposed to a 5‐mg kg−1 dose, whereas total protein, BUN and potassium levels significantly decreased. In conclusion, we suggest that the long persistence of GNP in the lung may cause adverse health effects by disturbing immunological‐ and physiological‐homeostasis of our body. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-07-21T05:50:40.280098-05:
      DOI: 10.1002/jat.3361
  • Hepatocytes cocultured with Sertoli cells in bioreactor favors Sertoli
           barrier tightness in rat
    • Authors: P. Zeller; A. Legendre, S. Jacques, M. J. Fleury, F. Gilard, G. Tcherkez, E. Leclerc
      Abstract: The lack of a reliable in vitro system to assess reprotoxicity is an emerging problem in the context of European law for Registration, Evaluation, Authorization and Restriction of Chemicals (REACH, 2007), as it requires a reduction in animal utilization for testing. Furthermore, in vitro reprotoxicological tests would be more relevant and greatly improved by integrating both hepatic metabolism and the blood–testis barrier. Here, we took advantage of an integrated insert in a dynamic microfluidic platform (IIDMP) to co‐cultivate hepatocytes in biochip and Sertoli cells in the bicameral chamber. This microfluidic tool has been previously demonstrated to be helpful in cell function and/or quality improvement. We demonstrate that permeability of the Sertoli barrier is reduced by dynamic coculture in our system. Exometabolomics analysis reveals that interactions between hepatocytes and Sertoli cells may have been mediated by the polyamines increase and/or mid‐chain fatty acid decrease in the circulating medium. These metabolic changes may be involved in permeability reduction by contributing to modifying junction protein quantity and localization. The present study gives an example of IIDMP as an in vitro partitioning/transport model for cell culture and toxicological testing. Further, based on both our previous results using an intestinal–hepatic cell coculture and the present study, IIDMP seems to be well‐suited for (i) assessing the dose–response effect of chemicals within the rodent or human male reproductive tract, and (ii) improving the quality of reprotoxicological assays by including hepatic metabolism. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-07-21T05:45:34.727774-05:
      DOI: 10.1002/jat.3360
  • In vitro toxicology studies of extracellular vesicles
    • Authors: Sayantan Maji; Irene K. Yan, Mansi Parasramka, Swathi Mohankumar, Akiko Matsuda, Tushar Patel
      Abstract: Extracellular vesicles (EVs) are membrane‐bound vesicles released from cells into the extracellular environment. There is emerging interest in the use of EVs as potential therapeutic interventions. We sought to evaluate the safety of EVs that may be therapeutically used by performing in vitro toxicological assessments. EVs were obtained from mesenchymal stem cells (MSC‐EV) or from bovine milk (BM‐EV) by differential ultracentrifugation, and quantitated using nanoparticle tracking analysis. Genotoxic effects, hematological effects, immunological effects and endotoxin production were evaluated at two dose levels. Neither MSC‐EVs nor BM‐EVs elicited detectable genotoxic effects using either the alkaline comet assay or micronucleus assay. Hemolysis was observed with BM‐EVs but not with MSC‐EVs. MSC‐EVs did not have any significant effect on either spontaneous or collagen‐induced platelet aggregation. In contrast, BM‐EVs were noted to increase collagen‐induced platelet aggregation, even though no spontaneous increase in platelet aggregation was noted. Both types of EVs induced leukocyte proliferation, which was greater with BM‐EV. Neither MSC‐EVs nor BM‐EVs induced HL‐60 phagocytosis, although BM‐EVs decreased zymosan‐induced phagocytosis. Furthermore, neither MSC‐EVs nor BM‐EVs induced nitric oxide production. Unlike MSC‐EVs, BM‐EVs tested positive for endotoxin and induced complement activation. There are significant differences in toxicological profiles between MSC‐EVs and BM‐EVs that may reflect variations in techniques for EV isolation, EV content or cross‐species differences. The safety of MSC‐EV supports their use for disease therapeutics, whereas detailed safety and toxicological assessment will be necessary before the use of BM‐EVs. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-07-20T05:20:51.296356-05:
      DOI: 10.1002/jat.3362
  • Molecular docking reveals the potential of phthalate esters to inhibit the
           enzymes of the glucocorticoid biosynthesis pathway
    • Authors: Shahzad Ahmad; Mohemmed Faraz Khan, Suhel Parvez, Mohammad Akhtar, Sheikh Raisuddin
      Abstract: Glucocorticoids (GCs) are well known to exert broad‐based effects on metabolism, behavior and immunity. Their impaired synthesis and production lead to adverse health effects. Some environmental toxicants, including phthalate esters (PAEs), are associated with endocrine disruption. These endocrine‐disrupting chemicals (EDCs) also cause adrenal toxicity and alteration of GC biosynthesis and their functions. Using in silico tools of Schrodinger Maestro 9.4, we performed a molecular docking study of 32 ligands including PAEs of a known endocrine‐disrupting potential with the selected enzymes of the GC biosynthesis pathway (GBP) such as CYP11A1, CYP11B2, CYP19A1, CYP17A1, CYP21A2 and 3α/20β‐HSD. Binding affinities of the PAEs were compared with known inhibitors of these enzymes. Amongst PAEs, diphenyl benzene‐1, 2 – dicarboxylate (DPhP) showed the lowest docking score of −8.95616 kcal mol−1 against CYP21A1. Besides, benzyl butyl benzene‐1,2‐dicarboxylate (BBzP), bis(7‐methylnonyl) benzene‐1,2 dicarboxylate (DIDP) and bis(2‐ethylhexyl) benzene‐1,2‐dicarboxylate (DEHP) also showed comparable molecular interaction with enzymes of GBP. DPhP showed a significant molecular interaction with different enzymes of GBP such as CYP21A1, CYP11A1 and CYP11B2. These interactions mainly included H‐bonding, hydrophobic, polar and van dar Waals' interactions. Interestingly, this in silico study revealed that certain PAEs have more inhibitory potential against enzymes of GBP than their respective known inhibitors. Such studies become more relevant in the risk assessment of exposure to mixtures of phthalate eaters. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-07-18T01:01:17.086181-05:
      DOI: 10.1002/jat.3355
  • Identification of microRNA biomarker candidates in urine and plasma from
           rats with kidney or liver damage
    • Authors: Francis S. Wolenski; Pooja Shah, Tomoya Sano, Tadahiro Shinozawa, Hugues Bernard, Matt J. Gallacher, Shylah D. Wyllie, Georgianna Varrone, Lisa A. Cicia, Mary E. Carsillo, Craig D. Fisher, Sean E. Ottinger, Erik Koenig, Patrick J. Kirby
      Abstract: MicroRNAs (miRNA) are short single‐stranded RNA sequences that have a role in the post‐transcriptional regulation of genes. The identification of tissue specific or enriched miRNAs has great potential as novel safety biomarkers. One longstanding goal is to associate the increase of miRNA in biofluids (e.g., plasma and urine) with tissue‐specific damage. Next‐generation sequencing (miR‐seq) was used to analyze changes in miRNA profiles of tissue, plasma and urine samples of rats treated with either a nephrotoxicant (cisplatin) or one of two hepatotoxicants (acetaminophen [APAP] or carbon tetrachloride [CCL4]). Analyses with traditional serum chemistry and histopathology confirmed that toxicant‐induced organ damage was specific. In animals treated with cisplatin, levels of five miRNAs were significantly altered in the kidney, 14 in plasma and six in urine. In APAP‐treated animals, five miRNAs were altered in the liver, 74 in plasma and six in urine; for CCL4 the changes were five, 20 and 6, respectively. Cisplatin treatment caused an elevation of miR‐378a in the urine, confirming the findings of other similar studies. There were 17 in common miRNAs elevated in the plasma after treatment with either APAP or CCL4. Four of these (miR‐122, −802, −31a and −365) are known to be enriched in the livers of rats. Interestingly, the increase of serum miR‐802 in both hepatotoxicant treatments was comparable to that of the well‐known liver damage marker miR‐122. Taken together, comparative analysis of urine and plasma miRNAs demonstrated their utility as biomarkers of organ injury. Copyright © 2016 The
      Authors . Journal of Applied Toxicology published by John Wiley & Sons Ltd.
      PubDate: 2016-07-11T03:20:51.930644-05:
      DOI: 10.1002/jat.3358
  • Anthophyllite asbestos: state of the science review
    • Authors: Shannon H. Gaffney; Matthew Grespin, Lindsey Garnick, Derek A. Drechsel, Rebecca Hazan, Dennis J. Paustenbach, Brooke D. Simmons
      Abstract: Anthophyllite is an amphibole form of asbestos historically used in only a limited number of products. No published resource currently exists that offers a complete overview of anthophyllite toxicity or of its effects on exposed human populations. We performed a review focusing on how anthophyllite toxicity was understood over time by conducting a comprehensive search of publicly available documents that discussed the use, mining, properties, toxicity, exposure and potential health effects of anthophyllite. Over 200 documents were identified; 114 contained relevant and useful information which we present chronologically in this assessment. Our analysis confirms that anthophyllite toxicity has not been well studied compared to other asbestos types. We found that toxicology studies in animals from the 1970s onward have indicated that, at sufficient doses, anthophyllite can cause asbestosis, lung cancer and mesothelioma. Studies of Finnish anthophyllite miners, conducted in the 1970s, found an increased incidence of asbestosis and lung cancer, but not mesothelioma. Not until the mid‐1990s was an epidemiological link with mesothelioma in humans observed. Its presence in talc has been of recent significance in relation to potential asbestos exposure through the use of talc‐containing products. Characterizing the health risks of anthophyllite is difficult, and distinguishing between its asbestiform and non‐asbestiform mineral form is essential from both a toxicological and regulatory perspective. Anthophyllite toxicity has generally been assumed to be similar to other amphiboles from a regulatory standpoint, but some notable exceptions exist. In order to reach a more clear understanding of anthophyllite toxicity, significant additional study is needed. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-07-11T03:05:57.857656-05:
      DOI: 10.1002/jat.3356
  • Is skin penetration a determining factor in skin sensitization potential
           and potency' Refuting the notion of a LogKow threshold for skin
    • Authors: Jeremy M. Fitzpatrick; David W. Roberts, Grace Patlewicz
      Abstract: It is widely accepted that substances that cannot penetrate through the skin will not be sensitizers. LogKow and molecular weight (MW) have been used to set thresholds for sensitization potential. Highly hydrophilic substances e.g. LogKow ≤ 1 are expected not to penetrate effectively to induce sensitization. To investigate whether LogKow >1 is a true requirement for sensitization, a large dataset of substances that had been evaluated for their skin sensitization potential under Registration, Evaluation, Authorisation and restriction of CHemicals (REACH), together with available measured LogKow values was compiled using the OECD eChemPortal. The incidence of sensitizers relative to non‐sensitizers above and below a LogKow of 1 was explored. Reaction chemistry principles were used to explain the sensitization observed for the subset of substances with a LogKow ≤0. 1482 substances were identified with skin sensitization data and measured LogKow values. 525 substances had a measured LogKow ≤ 1, 100 of those were sensitizers. There was no significant difference in the incidence of sensitizers above and below a LogKow of 1. Reaction chemistry principles that had been established for lower MW and more hydrophobic substances were found to be still valid in rationalizing the skin sensitizers with a LogKow ≤ 0. The LogKow threshold arises from the widespread misconception that the ability to efficiently penetrate the stratum corneum is a key determinant of sensitization potential and potency. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-06-29T20:40:33.379277-05:
      DOI: 10.1002/jat.3354
  • Toxicity of single‐wall carbon nanotubes functionalized with
           polyethylene glycol in zebrafish (Danio rerio) embryos
    • Authors: Felipe A. Girardi; Gisele E. Bruch, Carolina S. Peixoto, Lidiane Dal Bosco, Sangram K. Sahoo, Carla O. F. Gonçalves, Adelina P. Santos, Clascídia A. Furtado, Cristiano Fantini, Daniela M. Barros
      Abstract: Single‐wall carbon nanotubes functionalized with polyethylene glycol (SWCNT‐PEG) are promising materials for biomedical applications such as diagnostic devices and controlled drug‐release systems. However, several questions about their toxicological profile remain unanswered. Thus, the aim of this study was to investigate the action of SWCNT‐PEG in Danio rerio zebrafish embryos at the molecular, physiological and morphological levels. The SWCNT used in this study were synthesized by the high‐pressure carbon monoxide process, purified and then functionalized with distearoyl phosphatidylethanolamine block copolymer‐PEG (molecular weight 2 kDa). The characterization process was carried out with low‐resolution transmission electron microscopy, thermogravimetric analysis and Raman spectroscopy. Individual zebrafish embryos were exposed to the SWCNT‐PEG. Toxic effects occurred only at the highest concentration tested (1 ppm) and included high mortality rates, delayed hatching and decreased total larval length. For all the concentrations tested, the alkaline comet assay revealed no genotoxicity, and Raman spectroscopy measurements on the histological slices revealed no intracellular nanotubes. The results shown here demonstrate that SWCNT‐PEG has low toxicity in zebrafish embryos, but more studies are needed to understand what mechanisms are involved. However, the presence of residual metals is possibly among the primary mechanisms responsible for the toxic effects observed, because the purification process was not able to remove all metal contamination, as demonstrated by the thermogravimetric analysis. More attention must be given to the toxicity of these nanomaterials before they are used in biomedical applications. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-06-20T03:40:51.371492-05:
      DOI: 10.1002/jat.3346
  • Cytotoxic effects of psychotropic benzofuran derivatives,
           N‐methyl‐5‐(2‐aminopropyl)benzofuran and its N‐demethylated
           derivative, on isolated rat hepatocytes
    • Authors: Yoshio Nakagawa; Toshinari Suzuki, Yukie Tada, Akiko Inomata
      Abstract: The novel psychoactive compounds derived from amphetamine have been illegally abused as recreational drugs, some of which are known to be hepatotoxic in humans and experimental animals. The cytotoxic effects and mechanisms of 5‐(2‐aminopropyl)benzofuran (5‐APB) and N‐methyl‐5‐(2‐aminopropyl)benzofuran (5‐MAPB), both of which are benzofuran analogues of amphetamine, and 3,4‐methylenedioxy‐N‐methamphetamine (MDMA) were studied in freshly isolated rat hepatocytes. 5‐MAPB caused not only concentration‐dependent (0–4.0 mm) and time‐dependent (0–3 h) cell death accompanied by the depletion of cellular ATP and reduced glutathione and protein thiol levels, but also accumulation of oxidized glutathione. Of the other analogues examined at a concentration of 4 mm, 5‐MAPB/5‐APB‐induced cytotoxicity with the production of reactive oxygen species and loss of mitochondrial membrane potential was greater than that induced by MDMA. In isolated rat liver mitochondria, the benzofurans resulted in a greater increase in the rate of state 4 oxygen consumption than did MDMA, with a decrease in the rate of state 3 oxygen consumption. Furthermore, the benzofurans caused more of a rapid mitochondrial swelling dependent on the mitochondrial permeability transition than MDMA. 5‐MAPB at a weakly toxic level (1 mm) was metabolized slowly: levels of 5‐MAPB and 5‐APB were approximately 0.9 mm and 50 μm, respectively, after 3 h incubation. Taken collectively, these results indicate that mitochondria are target organelles for the benzofuran analogues and MDMA, which elicit cytotoxicity through mitochondrial failure, and the onset of cytotoxicity may depend on the initial and/or residual concentrations of 5‐MAPB rather than on those of its metabolite 5‐APB. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-06-13T01:10:49.273901-05:
      DOI: 10.1002/jat.3351
  • Detection of exposure effects of mixtures of heavy polycyclic aromatic
           hydrocarbons in zebrafish embryos
    • Authors: Alejandro Barranco; Laura Escudero, Jon Sanz Landaluze, Sandra Rainieri
      Abstract: In this study we evaluated the exposure effects of mixtures of five polycyclic aromatic hydrocarbons (PAHs); namely, benzo[a]anthracene, benzo[a]pyrene, benzo[b]fluoranthene, benzo[k]fluoranthene and chrysene on zebrafish embryos. Supplementation of the exposure media with 0.45% dimethyl sulfoxide and 50 ppm of Tween 20 could guarantee the solubilization and stabilization of the PAHs up to 24 h without affecting the embryos development. The exposure effects were tested by detecting the differential expression of a number of genes related to the aryl hydrocarbon receptor gene battery. Effects were detectable already after 6 h of exposure. After 24 h of exposure, all PAHs, except for benzo[a]anthracene, acted as potent inducers of the gene cyp1a1. Benzo[k]fluoranthene was the major inducer; the effect caused by the mixture at the lower concentration tested (1 ng ml−1) was dominated by its presence. However, in the mixture at the highest concentration tested (10 ng ml−1) it caused less induction and was not dominant. No significant bioaccumulation values were detected on embryos exposed to the PAHs tested in this study; however, the results obtained, indicated that PAHs undergo a very rapid metabolization inside the embryos, and that those biotransformation products yield changes on the expression of genes involved in the aryl hydrocarbon receptor pathway. Future work should focus on identification of the PAH metabolization products and on the effect of these metabolites on toxicity. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-06-10T01:25:31.717541-05:
      DOI: 10.1002/jat.3353
  • What determines skin sensitization potency: Myths, maybes and realities.
           The 500 molecular weight cut‐off: An updated analysis
    • Authors: Jeremy M. Fitzpatrick; David W. Roberts, Grace Patlewicz
      Abstract: It is widely accepted that substances must have a molecular weight (MW)  500, five were sensitizers. This provided good evidence to refute such a MW 500 threshold. While Roberts et al. (2012) made a convincing case that the MW > 500 cut‐off was not a true requirement for sensitization, the number of counter examples identified were too few to draw any statistical conclusions. This updated analysis systematically interrogated a large repository of sensitization information collected under the EU REACH regulation. A data set of 2904 substances that had been tested for skin sensitization, using guinea pigs and/or mice were collected. The data set contained 197 substances with a MW > 500; 33 of these were skin sensitizers. Metal containing complexes, reaction products and mixtures were excluded from further consideration. The final set of 14 sensitizers substantiated the original findings. The study also assessed whether the same reaction chemistry principles established for low MW sensitizers applied to chemicals with a MW > 500. The existing reaction chemistry considerations were found appropriate to rationalize the sensitization behaviour of the 14 sensitizers with a MW > 500. The existence of the MW 500 threshold, based on the widespread misconception that the ability to penetrate efficiently the stratum corneum is a key determinant of skin sensitization potential and potency, was refuted. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-06-10T01:15:33.619414-05:
      DOI: 10.1002/jat.3348
  • Comparative oral dose toxicokinetics of sodium selenite and
    • Authors: T. Zane Davis; Asheesh K. Tiwary, Bryan L. Stegelmeier, James A. Pfister, Kip E. Panter, Jeffery O. Hall
      Abstract: Selenium (Se) poisoning by different forms of Se occurs in the United States. However, the toxicokinetics of different selenocompounds after oral ingestion is not well documented. In this study the toxicokinetics of Se absorption, distribution and elimination were determined in serum and whole blood of lambs that were orally dosed with increasing doses of Se as sodium selenite (inorganic Se) or selenomethionine (SeMet, organic Se). Thirty‐two lambs were randomly assigned to eight treatment groups, with four animals per group. Se was administered at 1, 2 or 3 mg kg−1 body weight, as either sodium selenite or SeMet with proper control groups. Blood and serum were collected at predetermined time points for 7 days post‐dosing. Resulting Se concentrations in both serum and whole blood from SeMet treatment groups were significantly greater than those given equimolar doses of Se as sodium selenite. Se concentrations in serum and whole blood of lambs dosed with SeMet peaked at significantly greater concentrations when compared with lambs dosed with equimolar doses of sodium selenite. Based on the serum and whole blood kinetics, the rate of Se absorption was greater for SeMet than for sodium selenite although rates of absorption for both Se forms decreased with increasing dose. The rates of Se elimination increased with dose. These results demonstrate that SeMet has a greater absorption rate and a similar retention time resulting in a greater area under the curve and thus bioavailability than sodium selenite, which must be considered in both overdose and nutritional exposures. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.
      PubDate: 2016-06-10T01:05:25.404061-05:
      DOI: 10.1002/jat.3350
  • Preclinical safety study of a recombinant Streptococcus pyogenes vaccine
           formulated with aluminum adjuvant
    • Authors: Harm HogenEsch; Anisa Dunham, Elodie Burlet, Fangjia Lu, Yung‐Yi C. Mosley, Garry Morefield
      Abstract: A recombinant vaccine composed of a fusion protein formulated with aluminum hydroxide adjuvant is under development for protection against diseases caused by Streptococcus pyogenes. The safety and local reactogenicity of the vaccine was assessed by a comprehensive series of clinical, pathologic and immunologic tests in preclinical experiments. Outbred mice received three intramuscular injections of 1/5th of the human dose (0.1 ml) and rabbits received two injections of the full human dose. Control groups received adjuvant or protein antigen. The vaccine did not cause clinical evidence of systemic toxicity in mice or rabbits. There was a transient increase of peripheral blood neutrophils after the third vaccination of mice. In addition, the concentration of acute phase proteins serum amyloid A and haptoglobin was significantly increased 1 day after injection of the vaccine in mice. There was mild transient swelling and erythema of the injection site in both mice and rabbits. Treatment‐related pathology was limited to inflammation at the injection site and accumulation of adjuvant‐containing macrophages in the draining lymph nodes. In conclusion, the absence of clinical toxicity in two animal species suggest that the vaccine is safe for use in a phase I human clinical trial. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-05-30T21:05:44.533104-05:
      DOI: 10.1002/jat.3349
  • Lithium limits trimethyltin‐induced cytotoxicity and proinflammatory
           response in microglia without affecting the concurrent autophagy
    • Authors: Cinzia Fabrizi; Elena Pompili, Francesca Somma, Stefania De Vito, Viviana Ciraci, Marco Artico, Paola Lenzi, Francesco Fornai, Lorenzo Fumagalli
      Abstract: Trimethyltin (TMT) is a highly toxic molecule present as an environmental contaminant causing neurodegeneration particularly of the limbic system both in humans and in rodents. We recently described the occurrence of impairment in the late stages of autophagy in TMT‐intoxicated astrocytes. Here we show that similarly to astrocytes also in microglia, TMT induces the precocious block of autophagy indicated by the accumulation of the autophagosome marker, microtubule associated protein light chain 3. Consistent with autophagy impairment we observe in TMT‐treated microglia the accumulation of p62/SQSTM1, a protein specifically degraded through this pathway. Lithium has been proved effective in limiting neurodegenerations and, in particular, in ameliorating symptoms of TMT intoxication in rodents. In our in vitro model, lithium displays a pro‐survival and anti‐inflammatory action reducing both cell death and the proinflammatory response of TMT‐treated microglia. In particular, lithium exerts these activities without reducing TMT‐induced accumulation of light chain 3 protein. In fact, the autophagic block imposed by TMT is unaffected by lithium administration. These results are of interest as defects in the execution of autophagy are frequently observed in neurodegenerative diseases and lithium is considered a promising therapeutic agent for these pathologies. Thus, it is relevant that this cation can still maintain its pro‐survival and anti‐inflammatory role in conditions of autophagy block. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-05-25T23:20:33.492046-05:
      DOI: 10.1002/jat.3344
  • Toxic effects of chemical dispersant Corexit 9500 on water flea Daphnia
    • Authors: Kenji Toyota; Nicole A. McNabb, Demetri D. Spyropoulos, Taisen Iguchi, Satomi Kohno
      Abstract: In 2010, approximately 2.1 million gallons of chemical dispersants, mainly Corexit 9500, were applied in the Gulf of Mexico to prevent the oil slick from reaching shorelines and to accelerate biodegradation of oil during the Deepwater Horizon oil spill. Recent studies have revealed toxic effects of Corexit 9500 on marine microzooplankton that play important roles in food chains in marine ecosystems. However, there is still little known about the toxic effects of Corexit 9500 on freshwater zooplankton, even though oil spills do occur in freshwater and chemical dispersants may be used in response to these spills. The cladoceran crustacean, water flea Daphnia magna, is a well‐established model species for various toxicological tests, including detection of juvenile hormone‐like activity in test compounds. In this study, we conducted laboratory experiments to investigate the acute and chronic toxicity of Corexit 9500 using D. magna. The acute toxicity test was conducted according to OECD TG202 and the 48 h EC50 was 1.31 ppm (CIs 0.99–1.64 ppm). The reproductive chronic toxicity test was performed following OECD TG211 ANNEX 7 and 21 days LOEC and NOEC values were 4.0 and 2.0 ppm, respectively. These results indicate that Corexit 9500 has toxic effects on daphnids, particularly during the neonatal developmental stage, which is consistent with marine zooplankton results, whereas juvenile hormone‐like activity was not identified. Therefore, our findings of the adverse effects of Corexit 9500 on daphnids suggest that application of this type of chemical dispersant may have catastrophic impacts on freshwater ecosystems by disrupting the key food chain network. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-05-25T23:10:32.196468-05:
      DOI: 10.1002/jat.3343
  • Acetyl L‐carnitine targets adenosine triphosphate synthase in protecting
           zebrafish embryos from toxicities induced by verapamil and ketamine: An in
           vivo assessment
    • Authors: Xiaoqing Guo; Melanie Dumas, Bonnie L. Robinson, Syed F. Ali, Merle G. Paule, Qiang Gu, Jyotshna Kanungo
      Abstract: Verapamil is a Ca2+ channel blocker and is highly prescribed as an anti‐anginal, antiarrhythmic and antihypertensive drug. Ketamine, an antagonist of the Ca2+‐permeable N‐methyl‐d‐aspartate‐type glutamate receptors, is a pediatric anesthetic. Previously we have shown that acetyl l‐carnitine (ALCAR) reverses ketamine‐induced attenuation of heart rate and neurotoxicity in zebrafish embryos. Here, we used 48 h post‐fertilization zebrafish embryos that were exposed to relevant drugs for 2 or 4 h. Heart beat and overall development were monitored in vivo. In 48 h post‐fertilization embryos, 2 mm ketamine reduced heart rate in a 2 or 4 h exposure and 0.5 mm ALCAR neutralized this effect. ALCAR could reverse ketamine's effect, possibly through a compensatory mechanism involving extracellular Ca2+ entry through L‐type Ca2+ channels that ALCAR is known to activate. Hence, we used verapamil to block the L‐type Ca2+ channels. Verapamil was more potent in attenuating heart rate and inducing morphological defects in the embryos compared to ketamine at specific times of exposure. ALCAR reversed cardiotoxicity and developmental toxicity in the embryos exposed to verapamil or verapamil plus ketamine, even in the presence of 3,4,5‐trimethoxybenzoic acid 8‐(diethylamino)octyl ester, an inhibitor of intracellular Ca2+ release suggesting that ALCAR acts via effectors downstream of Ca2+. In fact, ALCAR's protective effect was blunted by oligomycin A, an inhibitor of adenosine triphosphate synthase that acts downstream of Ca2+ during adenosine triphosphate generation. We have identified, for the first time, using in vivo studies, a downstream effector of ALCAR that is critical in abrogating ketamine‐ and verapamil‐induced developmental toxicities. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.
      PubDate: 2016-05-18T07:05:10.31434-05:0
      DOI: 10.1002/jat.3340
  • Dibutyltin‐induced alterations of interleukin 1beta secretion from
           human immune cells
    • Authors: Shyretha Brown; Shahin Tehrani, Margaret M. Whalen
      Abstract: Dibutyltin (DBT) is used to stabilize polyvinyl chloride plastics (including pipes that distribute drinking water) and as a de‐worming agent in poultry. DBT is found in human blood, and DBT exposures alter the secretion of tumor necrosis factor alpha and interferon gamma from lymphocytes. Interleukin (IL)‐1β is a proinflammatory cytokine that regulates cellular growth, tissue restoration and immune response regulation. IL‐1β plays a role in increasing invasiveness of certain tumors. This study reveals that exposures to DBT (24 h, 48 h and 6 days) modify the secretion of IL‐1β from increasingly reconstituted preparations of human immune cells (highly enriched human natural killer cells, monocyte‐depleted [MD] peripheral blood mononuclear cells [PBMCs], PBMCs, granulocytes and a preparation combining both PBMCs and granulocytes). DBT altered IL‐1β secretion from all cell preparations. Higher concentrations of DBT (5 and 2.5 μm) decreased the secretion of IL‐1β, while lower concentrations of DBT (0.1 and 0.05 μm) increased the secretion of IL‐1β. Selected signaling pathways were examined in MD‐PBMCs to determine if they play a role in DBT‐induced elevations of IL‐1β secretion. Pathways examined were IL‐1β converting enzyme (caspase 1), mitogen‐activated protein kinases and nuclear factor kappa B. Caspase 1 and mitogen‐activated protein kinase pathways appear to be utilized by DBT in increasing IL‐1β secretion. These results indicate that DBT alters IL‐1β secretion from human immune cells in an ex. vivo system utilizing several IL‐1β regulating signaling pathways. Thus, DBT may have the potential to alter IL‐1β secretion in an in vivo system. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-05-17T01:35:28.9227-05:00
      DOI: 10.1002/jat.3339
  • Non‐clinical safety assessment of single and repeated administration of
           gE/AS01 zoster vaccine in rabbits
    • Authors: Giulia Giordano; Lawrence Segal, Menk Prinsen, Marcel V. W. Wijnands, Nathalie Garçon, Eric Destexhe
      Abstract: HZ/su is an investigational recombinant subunit vaccine for the prevention of shingles, a disease resulting from the reactivation of varicella zoster virus. The vaccine is composed of recombinant varicella zoster virus glycoprotein E (gE), and liposome‐based Adjuvant System AS01. To evaluate the potential local and systemic effects of this vaccine, three studies were performed in rabbits. In the first two studies, rabbits received a single intramuscular (IM; study 1) or subcutaneous (SC; study 2) dose of gE/AS01, AS01 alone (in study 2 only) or saline, and the local tolerance was evaluated up to 3 days after administration. Under these conditions, only local inflammatory reactions at the injection sites were detected by microscopic evaluation. In the third study, gE/AS01, AS01 alone or saline, were injected SC or IM on four occasions at 2 week intervals. General health status, local tolerance, ophthalmology, haematology and blood chemistry parameters were monitored. Macroscopic and microscopic evaluations were performed after termination of the study. The only treatment‐related changes included a transient increase in neutrophils, C‐reactive protein and fibrinogen levels and microscopic signs of inflammation at the injection sites, which are expected observations related to the elicited inflammatory reaction. The SC and IM routes of administration produced similar systemic effects. However, microscopic findings at the injection sites differed. One month after the last injection, recovery was complete in all groups. In conclusion, the single and repeated SC and IM administration of the gE/AS01 vaccine were locally and systemically well‐tolerated in rabbits and support the clinical development of the vaccine. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-05-12T08:03:54.99173-05:0
      DOI: 10.1002/jat.3329
  • 4‐Nitrophenol exposure alters the AhR signaling pathway and related gene
           expression in the rat liver
    • Authors: Ruonan Li; Meiyan Song, Zhi Li, Yansen Li, Gen Watanabe, Kentaro Nagaoka, Kazuyoshi Taya, Chunmei Li
      Abstract: 4‐Nitrophenol (PNP) is well known as an environmental endocrine disruptor. The aim of this study was to clarify the mechanism of PNP‐induced liver damage and determine the regulatory involvement of the aryl hydrocarbon receptor (AhR) signaling pathway and associated gene expression. Immature male Wistar–Imamichi rats (28 days old) were randomly divided into control and PNP groups, which consisted of 1‐ and 3‐day exposure (1 DE and 3 DE, respectively) and 3‐day exposure followed by 3‐day recovery (3 DE + 3 DR), groups. Each group was administered the vehicle or PNP (200 mg kg–1 body weight). The body and liver weight were significantly decreased in the 3 DE group. The mRNA expression levels of estrogen receptor‐α (ERα), glutathione S‐transferase (GST) and AhR exhibited a significant increase in the 1 DE group whereas, in contrast, that of cytochrome P450 (CYP) 1A1 decreased significantly in the 3 DE +3 DR group. AhR and CYP1A1 proteins were detected in the cytoplasm of hepatocytes of the 1 DE and 3 DE +3 DR groups whereas the ERα protein was found in the hepatocyte nuclei of the 1 DE and 3 DE groups. The present study demonstrates that PNP activated the AhR signaling pathway and regulated related CYP1A1 and GST gene expression in the liver. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-05-12T07:51:04.421728-05:
      DOI: 10.1002/jat.3332
  • Lack of genotoxic mechanisms in early‐stage furan‐induced
           hepatocellular tumorigenesis in gpt delta rats
    • Authors: Daisuke Hibi; Yu Yokoo, Yuta Suzuki, Yuji Ishii, Meilan Jin, Aki Kijima, Takehiko Nohmi, Akiyoshi Nishikawa, Takashi Umemura
      Abstract: Furan has been used as an intermediate in the chemical‐manufacturing industry and has been shown to contaminate various foods. Although furan induces hepatocellular tumors in rodents, equivocal results from in vitro and in vivo mutagenicity tests have caused controversy regarding the involvement of genotoxic mechanisms in furan‐induced carcinogenesis. In the present study, to elucidate the possible mechanisms underlying furan‐induced hepatocarcinogenesis, a comprehensive medium‐term analysis was conducted using gpt delta rats treated with furan at carcinogenic doses for 13 weeks. In the liver, the frequencies of gpt and Spi‐ mutants derived mainly from point and deletion mutations, respectively, were not changed, and there were no furan‐specific gpt mutations in furan‐treated rats. In contrast, the number and area of glutathione S‐transferase placental form (GST‐P)‐ positive foci were significantly increased in the high‐dose group. Also, the ratio of PCNA‐positive hepatocytes was significantly elevated in the same group, as supported by significant increases in cyclin d1 and cyclin e1 mRNA levels. Thus, it is highly probable that cell proliferation, but not genotoxic mechanisms, contribute to the development of GST‐P foci in furan‐treated rats. Based on the close relationship between GST‐P and neoplastic hepatocytes, these data allowed us to hypothesize that cell proliferation following signal transduction other than the mitogen‐activated protein kinase (MAPK)/ERK pathway may play a crucial role in early‐stage furan‐induced hepatocarcinogenesis. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-05-03T07:51:06.035242-05:
      DOI: 10.1002/jat.3331
  • Characterization of three human cell line models for high‐throughput
           neuronal cytotoxicity screening
    • Authors: Zhi‐Bin Tong; Helena Hogberg, David Kuo, Srilatha Sakamuru, Menghang Xia, Lena Smirnova, Thomas Hartung, David Gerhold
      Abstract: More than 75 000 man‐made chemicals contaminate the environment; many of these have not been tested for toxicities. These chemicals demand quantitative high‐throughput screening assays to assess them for causative roles in neurotoxicities, including Parkinson's disease and other neurodegenerative disorders. To facilitate high throughput screening for cytotoxicity to neurons, three human neuronal cellular models were compared: SH‐SY5Y neuroblastoma cells, LUHMES conditionally‐immortalized dopaminergic neurons, and Neural Stem Cells (NSC) derived from human fetal brain. These three cell lines were evaluated for rapidity and degree of differentiation, and sensitivity to 32 known or candidate neurotoxicants. First, expression of neural differentiation genes was assayed during a 7‐day differentiation period. Of the three cell lines, LUHMES showed the highest gene expression of neuronal markers after differentiation. Both in the undifferentiated state and after 7 days of neuronal differentiation, LUHMES cells exhibited greater cytotoxic sensitivity to most of 32 suspected or known neurotoxicants than SH‐SY5Y or NSCs. LUHMES cells were also unique in being more susceptible to several compounds in the differentiating state than in the undifferentiated state; including known neurotoxicants colchicine, methyl‐mercury (II), and vincristine. Gene expression results suggest that differentiating LUHMES cells may be susceptible to apoptosis because they express low levels of anti‐apoptotic genes BCL2 and BIRC5/survivin, whereas SH‐SY5Y cells may be resistant to apoptosis because they express high levels of BCL2, BIRC5/survivin, and BIRC3 genes. Thus, LUHMES cells exhibited favorable characteristics for neuro‐cytotoxicity screening: rapid differentiation into neurons that exhibit high level expression neuronal marker genes, and marked sensitivity of LUHMES cells to known neurotoxicants. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-05-03T07:44:26.906766-05:
      DOI: 10.1002/jat.3334
  • Regucalcin counteracts tert‐butyl hydroperoxide and cadmium‐induced
           oxidative stress in rat testis
    • Authors: Sara Correia; Cátia V. Vaz, Ana M. S. Silva, José E. Cavaco, Sílvia Socorro
      Abstract: Regucalcin (RGN) is a calcium (Ca2+)‐binding protein with multiple physiological roles and has also been linked to the suppression of oxidative stress. It is widely known that oxidative stress adversely affects spermatogenesis, disrupting the development of germ cells, and interfering with sperm function. The present study aims to analyze the role of RGN modulating testicular oxidative stress. To address this issue, seminiferous tubules (SeT) from transgenic rats overexpressing RGN (Tg‐RGN) and wild‐type (WT) were cultured ex vivo for 24 h in the presence/absence of pro‐oxidant stimuli, tert‐butyl hydroperoxide (TBHP, 250 and 500 μM) and cadmium chloride (Cd, 10 and 20 μM). Noteworthy, SeT from Tg‐RGN animals displayed a significantly higher antioxidant capacity and diminished levels of thiobarbituric acid reactive substances relatively to their WT counterparts, both in control and experimental conditions. Regarding the antioxidant defense systems, a significant increase in the activity of glutathione‐S‐transferase was found in the SeT of Tg‐RGN whereas no differences were observed in superoxide dismutase activity throughout experimental conditions. The activity of apoptosis executioner caspase‐3 was significantly increased in the SeT of WT rats treated with 250 μM of TBHP or 10 μM of Cd, an effect not seen in Tg‐RGN animals. These results showed that the SeT of Tg‐RGN animals displayed lower levels of oxidative stress and increased antioxidant defenses, exhibiting protection against oxidative damage and apoptosis. Moreover, the present findings support the antioxidant role of RGN in spermatogenesis, which may be an important issue of further research in the context of male infertility. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-04-25T02:46:48.66054-05:0
      DOI: 10.1002/jat.3333
  • Neuroglial alterations in the zebrafish brain exposed to cadmium chloride
    • Authors: Antonio Monaco; Maria C. Grimaldi, Ida Ferrandino
      Abstract: Cadmium is an extremely toxic heavy metal that widely occurs in industrial workplaces with various hazardous effects on brain functions. The cytotoxic effects of cadmium chloride (CdCl2) on the neuroglial components of the zebrafish brain were analysed by detecting the glial fibrillary acidic protein (GFAP) expression and the mRNA levels of myelin genes mbp, mpz and plp1 in adult specimens exposed to cadmium for 2, 7 and 16 days. A significant decrease in the GFAP protein by Western blotting experiments was observed after 2 days of treatment, reaching 55% after 16 days. No change was observed in the mRNA levels. Using immunohistochemistry, a reduction in GFAP‐positive structures was revealed with a progressive trend in all the brains at 2, 7 and 16 days of treatment. In particular, a considerable reduction in GFAP‐positive fibres, with a different course, was observed in the ventricle areas and at the pial surface and in blood vessels after 16 days. Our experiments also showed a structural and chemical alteration of myelin and upregulation of mpz mRNA levels, the oligodendrocyte gene that is upregulated in experiments of neuronal injury, but not of plp1 and mbp mRNA levels, other myelin structural genes. These data confirm the toxic action of cadmium on the zebrafish brain. This action is time‐dependent and involves the glial cells, key components of the protection and function of nerve cells, hence the basis for many neurological diseases. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-04-15T04:01:49.760995-05:
      DOI: 10.1002/jat.3328
  • Comparison of Screening Dilution and Automated Reading for Antinuclear
           Antibody Detection on HEP2 Cells in the Monitoring of Connective Tissue
    • Authors: Anne E. Depincé-Berger; Amelie Moreau, Virginie Bossy, Christian Genin, Melanie Rinaudo, Stephane Paul
      Pages: 471 - 478
      Abstract: BackgroundIndirect immunofluorescence plays a major role in the detection of antinuclear antibodies (ANAs) and follow-up of their titers in the context of connective tissue diseases. Given the numerous unfavorable features of the conventional manual reading of HEP2 slides (need of time and expert morphologists for the reading, lack of standardization, subjectivity of the interpretation), the biomedical industry has developed automated techniques of slide preparation and microscope reading.MethodsWe collected 49 sera beforehand analyzed by the conventional reading of slides. They were prepared again by QUANTA-Lyser® and reanalyzed in four different conditions: two dilutions of screening (1/40 and 1/80), two different systems of analysis, NOVA View® automated reading (INOVA Diagnostics), then confirmation by the operator, and conventional manual reading by two different qualified operators. The analysis was realized in blind of the first interpretation and clinical diagnosis. The sera were classified in four groups, on the basis of the results of the first analysis: negative sera (titer < 1/160; 11 patients), low positives (titer at 1/160; 18 patients), moderated positives (titers between 1/320 and 1/640; 10 patients), and strong positives (titers between 1/1,280 and 1/2,560; 10 patients).ResultsAmong the 49 patients, 13 presented a connective tissue disease including 4 systemic scleroderma (SS), 3 rheumatoid arthritis (RA), 2 Goujerot-Sjogren (GS), 2 systemic lupus erythematosus (SLE), 1 polymyositis (PM), 1 Raynaud's syndrome (RS), and 1 CREST syndrome. One patient presented both an SLE and an SS.Regarding the screening dilution, the 1/40 dilution is less specific than the 1/80 dilution for both the systems of analysis (5.6% vs. 16.7% for the manual reading, and 27.8% vs. 50% for the automated reading). It also generates statistically more false positives (P = 0.037 for the conventional analysis and P = 0.003 for the automated system). The automated NOVA View® reading of slides allows a gain in specificity for both dilutions, and also statistically less false positives (P = 0.002 at the 1/40 and P = 0.0006 at the 1/80), and detriment of the sensitivity at the highest dilution (84.6% vs. 92.3% with manual reading). Thus, according to our analysis of 49 sera, the automated NOVA View® system of reading of slides at the dilution 1/80 seems to be a successful condition for the detection of ANAs on HEP2 cells, close to the significance (P = 0.067).ConclusionThe automated NOVA View® reading of slides allows saving time, and an improvement in the standardization. Nevertheless, it requires a confirmation by a qualified operator, to interpret mixed patterns in particular.
      PubDate: 2016-05-26T04:00:35.38806-05:0
      DOI: 10.1002/jcla.21881
  • Fecal Calprotectin: Diagnostic Accuracy of the Immunochromatographic
           CalFast Assay in a Pediatric Population
    • Authors: Oriano Radillo; Lorella Pascolo, Stefano Martelossi, Sara Dal Bo, Alessandro Ventura
      Pages: 500 - 505
      Abstract: BackgroundFecal calprotectin is a noninvasive marker for bowel diseases and it is high valuable to follow disease activity in Crohn's disease (CD) and ulcerative colitis (UC). In this study, we evaluated the diagnostic performance of the recently introduced immunochromatographic assay CalFast in comparison to the well-known ELISA tests for calprotectin assay to obtain a rapid diagnosis of bowel inflammation in pediatric patients.MethodsCalFast was tested in parallel to the classic ELISA tests CalPrest and PhiCal (gold standards for the calprotectin determination) on 148 fecal samples from pediatric subjects including 104 healthy subjects, 29 with CD, and 15 with UC.ResultsIn this study, the sensitivity and specificity of CalFast, CalPrest, and PhiCal were 86.4%, 88.6%, and 93.2% and 86.6%, 74%, and 64.4%, respectively. The area under the curve, obtained from receiver operating characteristic analysis, indicated the lack of significant difference among all the kits used.ConclusionThe immunochromatographic assay demonstrated good diagnostic predictive values, comparable to those of the ELISA methods, and may represent a valid alternative in order to save operators' time. The test, in fact, has a short turnaround time and does not need a specific ELISA instrumentation.
      PubDate: 2016-02-15T22:46:39.894851-05:
      DOI: 10.1002/jcla.21886
  • Antifungal Activity of Propolis Against Yeasts Isolated From Blood
           Culture: In Vitro Evaluation
    • Authors: Fatma Mutlu Sariguzel; Elife Berk, Ayes Nedret Koc, Hafize Sav, Gonca Demir
      Pages: 513 - 516
      Abstract: Background: Due to the failure of available antifungal agents in the treatment of candidemia and the toxic activities of these drugs, a lot of researches are being conducted to develop new nontoxic and effective antifungal agents for optimal control of fungal pathogens. The aim of this study is to evaluate the in vitro antifungal activity of propolis against yeasts isolated from the blood cultures of intensive care unit patients.Methods: Seventy-six strains were included in this study. The in vitro antifungal activity of propolis, fluconazole (FLU), and itraconazole (ITR) was investigated by the microdilution broth methods (CLSI guidelines M27-A3 for yeast). The propolis sample was collected from Kayseri, Turkey.Results: Of the 76 isolates, 33 were identified as Candida albicans while 37 were C. parapsilosis, three were C. tropicalis, and three were identified as C. glabrata. The geometric mean range for MIC (μg/ml) with regard to all isolates was 0.077 to 3 μg/ml for FLU and ITR, and 0.375 to 0.70 μg/ml for propolis. It was shown that propolis had significant antifungal activity against all Candida strains and the MIC range of propolis was determined as 0185 to 3 μg/ml.Conclusion: This study demonstrated that propolis had significant antifungal activity against yeasts isolated from blood culture compared with FLU and ITR. The propolis MIC in azole-resistant strains such as C. glabrata was found lower than the FLU MIC.
      PubDate: 2016-01-20T23:30:41.422046-05:
      DOI: 10.1002/jcla.21889
  • Stability of Routine Biochemical Analytes in Whole Blood and Plasma From
           Lithium Heparin Gel Tubes During 6-hr Storage
    • Authors: Denis Monneret; Alexandre Godmer, Ronan Guen, Clotilde Bravetti, Cecile Emeraud, Anthony Marteau, Rana Alkouri, Fouzi Mestari, Sylvie Dever, Françoise Imbert-Bismut, Dominique Bonnefont-Rousselot
      Pages: 602 - 609
      Abstract: BackgroundThe stability of biochemical analytes has already been investigated, but results strongly differ depending on parameters, methodologies, and sample storage times. We investigated the stability for many biochemical parameters after different storage times of both whole blood and plasma, in order to define acceptable pre- and postcentrifugation delays in hospital laboratories.MethodsTwenty-four analytes were measured (Modular® Roche analyzer) in plasma obtained from blood collected into lithium heparin gel tubes, after 2–6 hr of storage at room temperature either before (n = 28: stability in whole blood) or after (n = 21: stability in plasma) centrifugation. Variations in concentrations were expressed as mean bias from baseline, using the analytical change limit (ACL%) or the reference change value (RCV%) as acceptance limit.ResultsIn tubes stored before centrifugation, mean plasma concentrations significantly decreased after 3 hr for phosphorus (–6.1% [95% CI: –7.4 to –4.7%]; ACL 4.62%) and lactate dehydrogenase (LDH; –5.7% [95% CI: –7.4 to –4.1%]; ACL 5.17%), and slightly decreased after 6 hr for potassium (–2.9% [95% CI: –5.3 to –0.5%]; ACL 4.13%). In plasma stored after centrifugation, mean concentrations decreased after 6 hr for bicarbonates (–19.7% [95% CI: –22.9 to –16.5%]; ACL 15.4%), and moderately increased after 4 hr for LDH (+6.0% [95% CI: +4.3 to +7.6%]; ACL 5.17%). Based on RCV, all the analytes can be considered stable up to 6 hr, whether before or after centrifugation.ConclusionThis study proposes acceptable delays for most biochemical tests on lithium heparin gel tubes arriving at the laboratory or needing to be reanalyzed.
      PubDate: 2016-02-18T21:08:20.94555-05:0
      DOI: 10.1002/jcla.21909
  • Comparison of Molecular, Microscopic, and Culture Methods for Diagnosis of
           Cutaneous Leishmaniasis
    • Authors: Sima Rasti; Baharak Ghorbanzadeh, Farnaz Kheirandish, Seyed Gholamabbas Mousavi, Ahmad Pirozmand, Hossein Hooshyar, Bathol Abani
      Pages: 610 - 615
      Abstract: Cutaneous leishmaniasis (CL) is endemic in the northwest of Isfahan province, Iran. Increase in the incidence of the disease in Kashan has made it necessary to find out the best method for diagnosis and molecular characterization of Leishmania species. In the present study, 130 patients suspected to cutaneous leishmaniosis referred to health care centers of Kashan were examined. Serosity of lesion was collected for smear preparation and cultured in Novy-Nicolle-McNeal medium. DNA was extracted from serosity, and Leishmania species was determined by polymerase chain reaction (PCR) and nested PCR using kinetoplast DNA (kDNA) specific primers. The diagnostic criteria of CL were based on the observation of amastigotes in the smear, promastigotes in culture, presence of expected bands in PCR, or nested PCR. Of 130 specimens, 87 (66.9%), 72 (56.2%), 98 (75.4 %), 96 (73.8%), and 99 (76.2%) were positive for microscopic culture, PCR, nested PCR, and combined PCR and microscopy (proposed method), respectively. Sensitivity, specificity, positive and negative predictive values of PCR were 99%, 100%, 100%, 96.9%, respectively, for microscopy 87.9%, 100%, 100%, 72.1%, for culture 72.7%, 100%, 100%, 53.4 %, and for nested PCR 97%, 100%, 100%, 91.2%, respectively. Based on the results of the study, kDNA-PCR was the most sensitive method for diagnosis of CL.
      PubDate: 2016-02-18T21:06:54.603838-05:
      DOI: 10.1002/jcla.21910
  • Circulating Tumor Cells and Tumor Stem Cells Detection in the Peripheral
           Blood Mononuclear Cells of Breast Cancer
    • Authors: Feng Wang; Yuan-Chun Li, Li-Ping Liu, Hao-Min Zhang, Song Tong
      Pages: 616 - 622
      Abstract: BackgroundOur aim was to retrospectively analyze the relationships between circulating tumor cells (CTCs) and the development of breast cancer, for elucidating the role of CTCs in breast cancer.MethodsA total of 107 female patients with primary breast cancer and 48 matched healthy female volunteers were recruited. After blood collection, isolation of peripheral blood mononuclear cells (PBMC) was performed followed by the detection of cytokeratin 19 positive (CK19+) and CD44+/CD24−/low cells, as well as estrogen receptor (ER), progesterone, and CerbB2. Data were analyzed with the SPSS 20.0 software.ResultsNone of the 48 volunteers were detected with CK19+ cells in their PBMC, while in 77 patients, 72% of 107 female patients with primary breast cancer, the CK19+ cells were detected. CK19+ could also be detected among patients in each grouping by different clinical staging and lymph node metastasis, with statistical differences (all P < 0.05). Further, among the 83 CK19+ specimens, 32 were also detected with CD44+/CD24−/low cells. Comparisons of CK19+ and CD44+/CD24−/low cells in patients with different clinical features (ER positive vs. ER negative, C-erbB2 positive vs. C-erbB2 negative) and molecular subtypes (triple-negative breast cancer, ER positive, and C-erbB2 positive) showed no obvious difference (all P > 0.05).ConclusionsBoth CTCs and tumor stem cells (TSCs) could be detected in the PBMC of breast cancer patients; besides, positive expression rate of CTCs might be obviously associated with the clinical stage and metastasis. Positive relationship of TSCs and the clinical stage of breast cancer was also proved in this study.
      PubDate: 2016-02-18T21:06:34.338196-05:
      DOI: 10.1002/jcla.21911
  • Clinical Value of Hypochromia Markers in the Detection of Latent Iron
           Deficiency in Nonanemic Premenopausal Women
    • Authors: Eloísa Urrechaga; Luís Borque, Jesús F. Escanero
      Pages: 623 - 627
      Abstract: BackgroundIron deficiency (ID) is the most common cause of anemia in fertile women and hemoglobin (Hb) within the reference interval does not exclude ID. The consequence of an imbalance between the iron requirements and supply is a reduction of red-cell Hb content, which causes hypochromic cells. The aim of this study was to assess the reliability of new parameters low Hb density (LHD%), reticulocyte Hb equivalent (RetHe), and percentage of hypochromic erythrocytes (%HypoHe) in the detection of latent ID (LID), defined as depletion of iron stores without anemia.MethodsTwo hundred fifty consecutive nonanemic women in fertile age (18–40 years, mean 33.5 years), whose analyses had been requested by general practitioners, were included. Independent samples t-test, receiver operating characteristic (ROC) curve analysis (gold standard for detecting LID ferritin
      PubDate: 2016-02-22T01:02:42.518502-05:
      DOI: 10.1002/jcla.21912
  • Detection of Treponema pallidum Sp. Pallidum DNA in Cerebrospinal Fluid
           (CSF) by Two PCR Techniques
    • Authors: Rita Castro; Maria João Águas, Teresa Batista, Carlos Araújo, Kamal Mansinho, Filomena da Luz Martins Pereira
      Pages: 628 - 632
      Abstract: BackgroundLaboratory diagnosis of neurosyphilis is complicated especially when it is asymptomatic, no single laboratory test result being appropriate to diagnose central nervous system infectivity caused by Treponema pallidum. Our objective was to evaluate two polymerase chain reaction (PCR) techniques for the detection of T. pallidum DNA in the cerebrospinal fluid (CSF) of patients with syphilis.MethodsOne hundred twenty-four CSF samples from patients with reactive blood tests for syphilis were obtained. Two PCR techniques (47-PCR, polA-PCR) were used to detect T. pallidum DNA. The laboratory criteria used for the diagnosis of neurosyphilis to which the PCR techniques were compared were those recommended by the IUSTI: 2008 European guidelines on the management of syphilis.ResultsTreponema pallidum DNA was detected amplified in 37 of 124 (29.8%) and 30 of 124 (24.2%) samples with the 47-PCR and polA-PCR, respectively. Sensitivities were 75.8% and 69.7% and specificities 86.8% and 92.3%, respectively, for 47-PCR and polA-PCR techniques, respectively. The three CSF samples of patients with primary syphilis did not fulfill the criteria of neurosyphilis and DNA was only detected in one by the 47-PCR. In samples from secondary syphilis and neurosyphilis, three of nine and nine of nine respectively, results were coincident for the two PCR techniques and neurosyphilis criteria. Major discrepancies between the two PCR techniques and neurosyphilis diagnostic criteria were observed in latent syphilis.ConclusionBeyond some limitations of the study, which are discussed here, both PCR techniques seem to be useful for the diagnosis of neurosyphilis, although 47-PCR presents a higher sensitivity and polA-PCR a higher specificity.
      PubDate: 2016-02-18T21:05:49.681991-05:
      DOI: 10.1002/jcla.21913
  • Scanning for α-Hemoglobin Variants by High-Resolution Melting
    • Authors: Walaiporn Yimniam; Sumalee Jindadamrongwech
      Pages: 633 - 640
      Abstract: BackgroundDefinitive detection of hemoglobin (Hb) variants requires DNA sequencing. High-resolution melting (HRM) analysis of polymerase chain reaction (PCR) amplicons was applied to detect and discriminate among uncommon α-Hb variants found in Thailand.MethodsUncommon suspected α-Hb variants observed in Hb typing were identified by sequencing of DNA from whole blood samples. Three pairs of PCR primers covering the mutation regions in the three α-globin exons then were used for PCR coupled with difference in HRM analysis to subtract out the concomitant melting profile of the normal allele in the heterozygous state.ResultsDNA sequencing identified six heterozygous α-Hb variants, namely, Hb G-Waimanalo (HBA2: exon 2, codon 64 G>A), Hb J-Buda (HBA1: exon 2, codon 61 G>T), Hb Kurosaki (HBA2: exon 1; codon 7 A>G), Hb O-Indonesia (HBA1: exon 3 codon 116 G>A), Hb Q-India (HBA1:exon 2, codon 64 G>C), and Hb Q-Thailand (HBA1: exon 2 codon 74 G>C). Difference HRM analysis showed one temperature melting profile using exon 1 primer pair, four different profiles with exon 2 primer pair, and one profile with exon 3 primer pair.ConclusionsPCR-HRM analysis was effective in detecting and discriminating among single point mutations causing six uncommon α-Hb variants in heterozygous individuals. The method can be applied for routine screening due to its simplicity and relatively low cost.
      PubDate: 2016-02-18T21:07:59.015325-05:
      DOI: 10.1002/jcla.21914
  • Is Turkish MEFV Mutations Spectrum Different Among Regions?
    • Authors: Gulsen Yilmaz; Mehmet Senes, Damla Kayalp, Dogan Yucel
      Pages: 641 - 644
      Abstract: BackgroundFamilial Mediterranean fever (FMF) is an autosomal recessive inherited inflammatory disease. The gene responsible for the disease, called MEFV, encodes a protein called pyrin or marenostrin. According to recent data, MEFV mutations are not the only cause of FMF, but genetic analysis of MEFV gene is needed for confirming the diagnosis of FMF. In the present study, we aimed to evaluate the molecular testing results of MEFV mutations.MethodsMolecular testing results of 1,435 patients were retrospectively evaluated over the last 4 years. These patients were identified as having FMF clinical symptoms. Patients were tested for 12 common mutations in the MEFV gene using a strip assay technique.ResultsFrom all 1,435 patients, MEFV mutations were found in 776 patients (54.08%) and 659 patients (45.92%) did not carry any mutations. Patients with mutations were classified as homozygotes (n = 148), compound heterozygotes (n = 197), heterozygous (n = 427), and complex genotypes (n = 4, patients with three mutations). Allelic frequencies for the four most common mutations in the mutation-positive groups were 48.79% (M694V), 14.86% (M680I G/C), 13.70% (E148Q), and 12.35% (V726A). The remaining alleles (10.3%) showed rare mutations that were R761H, P369S, A744S, K695R, F479L, and M694I. No patient showed a I692del mutation that is sometimes evident in other Mediterranean populations.ConclusionIt was found that the most common four mutations (M694V, M680I [G/C], E148Q, V726A) were similar to those previously reported from different regions of Turkey and this study might add some knowledge to the mutational spectrum data on FMF.
      PubDate: 2016-02-18T21:01:03.758085-05:
      DOI: 10.1002/jcla.21915
  • Reference Intervals of Fibrosis Index Based on Four Indicators in Healthy
           Elderly Chinese
    • Authors: Guo-Ming Zhang; Yong-Jie Xia
      Pages: 645 - 648
      Abstract: BackgroundThe establishment of reference intervals of total bilirubin, Alanine aminotransferase (ALT), aspartate aminotransferases (AST), and creatinine provided necessary reference in screening and diagnosis of various kidney and liver diseases. However, these reference intervals were not available to estimate liver fibrosis degree. The purpose of this study is to establish the reference intervals of fibrosis index based on the four indicators (FIB-4) in apparently healthy elderly Chinese.MethodsA total of 24,949 blood specimens were collected by the standard procedures, and ALT, AST, and PLT were determined. FIB-4 were calculated by the following formula: FIB-4 = (age [years] × AST [U/l])/((PLT [109/l]) × (ALT [U/l])1/2).The elderly's FIB-4 were analyzed between the same age of different sexes and different ages of the same sex. Statistical data were analyzed by SPSS18.0 software.ResultsReference intervals of FIB-4 index, established for the healthy elderly, were 0.9923–4.5424 for males and 0.9007–4.1934 for females.ConclusionWe established reference intervals of FIB-4 index. This research provided reference value that can be used by relevant clinicians and inspection officers.
      PubDate: 2016-03-08T02:09:18.78109-05:0
      DOI: 10.1002/jcla.21916
  • Correlation of Serum and Salivary Cytokines Level With Clinical Parameters
           in Metabolic Syndrome With Periodontitis
    • Authors: Abhishek Chauhan; Suraj Singh Yadav, Pradeep Dwivedi, Nand Lal, Kauser Usman, Sanjay Khattri
      Pages: 649 - 655
      Abstract: BackgroundMetabolic Syndrome (MS) and chronic oral condition (periodontitis [PD]) are state of inflammation. The study was conducted to determine alterations in serum and salivary cytokines level in MS and/or chronic PD in the North Indian population.Materials and MethodsThis cross-sectional study carried out in northern part of India. The study subjects of similar ethnicity were recruited according to International Diabetes Federation (IDF) criteria for MS, while chronic PD was diagnosed on the basis of packet depth and clinical attachment level. ELISA method was employed to assess cytokine level. All subjects were divided in four groups Gr A (MS + PD), B (MS), C (PD), and a control Gr D.ResultsThe serum and salivary tumor necrosis factor alpha (TNF-α) level in Gr A, B, and C was significantly higher than Gr D (P 
      PubDate: 2016-02-22T05:16:48.26425-05:0
      DOI: 10.1002/jcla.21917
  • Detection of Circulating Tumor Cells Using a Novel Immunomagnetic Bead
           Method in Lung Cancer Patients
    • Authors: Jin-Ling Ji; Yu-Zhang Jiang, Qian-Qiu Tang, Xiao-Dong He, Zuo-Jun Shen, Bai-Yin Zhang
      Pages: 656 - 662
      Abstract: BackgroundCirculating tumor cells (CTCs) are detectable in peripheral blood of metastatic lung cancer patients. In this article, we evaluate a new CTC separation method based on a combination of anti-EpCAM and immunomagnetic beads with the aim to detect CTCs more conveniently and specifically.MethodsLung cancer cells were magnetically labeled by anti-EpCAM magnetic beads, and subsequently captured by magnetic separation using our novel device. Isolated lung cancer cells were identified by pathomorphological by hematoxylin–eosin staining protocol. The system was used to detect CTCs in 2 ml blood. Blood samples of healthy donors spiked with lung cancer cell line A549 cells were used to determine the sensitivity and specificity of the method. Prevalence of CTCs was examined in samples from 56 patients with lung cancer.ResultsRegression analysis of number of recovered versus spiked A549 cells yielded a coefficient of determination of R2 = 0.996 (P < 0.001). The average recovery was 68% or more at each spiking level. The coefficient of variation increased as the number of spiked cells decreased, ranging from 6.4% (1,000-cell spike) to 18.4% (50-cell spike). Forty-nine of the fifty-six patients (87.5%) were found to have CTCs in peripheral blood. None of the 2 ml peripheral blood samples of the 20 healthy subjects analyzed were found to have CTCs.ConclusionsThis novel turbulence device provides a new tool allowing for feasible and specific detection of CTCs in lung cancer patients. It is likely clinically useful in diagnosis and monitoring of lung cancer and may have a role in clinical decision making.
      PubDate: 2016-03-14T07:16:20.43789-05:0
      DOI: 10.1002/jcla.21918
  • Comparison of the Automated cobas u 701 Urine Microscopy and UF-1000i Flow
           Cytometry Systems and Manual Microscopy in the Examination of Urine
    • Authors: Wonmok Lee; Jung-Sook Ha, Nam-Hee Ryoo
      Pages: 663 - 671
      Abstract: BackgroundThe cobas u 701, a new automated image-based urine sediment analyzer, was introduced recently. In this study, we compared its performance with that of UF-1000i flow cytometry and manual microscopy in the examination of urine sediments.MethodsPrecision, linearity, and carry-over were determined for the two urine sediment analyzers. For a comparison of the method, 300 urine samples were examined by the automated analyzers and by manual microscopy using a KOVA chamber.ResultsWithin-run coefficients of variation (CVs) for the control materials were 7.0–8.8% and 1.7–5.7% for the cobas u 701 and UF-1000i systems, respectively. Between-run CVs were 8.5–9.8% and 2.7–5.4%, respectively. Both instruments showed good linearity and negligible carry-over. For red blood cells (RBC), white blood cells (WBC), and epithelial cells (EPI), the overall concordance rates within one grade of difference among the three methods were good (78.6–86.0%, 88.7–93.8%, and 81.3–90.7%, respectively). The concordance rate for casts was poor (66.5–68.9%).ConclusionCompared with manual microscopy, the two automated sediment analyzers tested in this study showed satisfactory analytical performances for RBC, WBC, and EPI. However, for other urine sediment particles confirmation by visual microscopy is still required.
      PubDate: 2016-02-03T20:40:50.609694-05:
      DOI: 10.1002/jcla.21919
  • Association of Serum hs-CRP Levels With the Presence of Obesity, Diabetes
           Mellitus, and Other Cardiovascular Risk Factors
    • Authors: Mahmoud Ebrahimi; Ali Reza Heidari-Bakavoli, Sara Shoeibi, Seyed Reza Mirhafez, Mohsen Moohebati, Habibollah Esmaily, Hamed Ghazavi, Maryam Saberi Karimian, Seyed Mohammad Reza Parizadeh, Maryam Mohammadi, Hossein Mohaddes Ardabili, Gordon A. Ferns, Majid Ghayour-Mobarhan
      Pages: 672 - 676
      Abstract: BackgroundDiabetes mellitus remains one of the major health problems of the 21st century and is associated with comorbidities including obesity and metabolic abnormalities. The study was conducted to evaluate serum high-sensitivity C-reactive protein (hs-CRP) levels, as a marker of inflammation, in a large sample of Iranian population without a history of cardiovascular or inflammatory disease and cancer, and to relate this to fasting blood glucose (FBG) and the presence of diabetes mellitus.MethodsThe study consisted of 7,762 subjects divided into four groups—nonobese/nondiabetic, obese/nondiabetic, nonobese/diabetic and obese/diabetic—based on the BMI classification and their FBG. Anthropometric characteristics were measured and blood was collected for the evaluation of fasted lipid profile, FBG and serum hs-CRP levels.ResultsSeveral clinical and biochemical characteristics were significantly different among the four groups: FBG, P < 0.001; total cholesterol (TC), P < 0.001; and triglyceride (TG), P < 0.001. The subjects with a serum hs-CRP >3 mg/dl had higher TC (P < 0.001), low-density lipoprotein cholesterol (LDL-C, P < 0.001), TG (P < 0.001), fat percentage (P < 0.001), and systolic and diastolic blood pressure (P < 0.001) compared with subjects with a serum hs-CRP
      PubDate: 2016-02-08T23:01:39.246382-05:
      DOI: 10.1002/jcla.21920
  • The Roche Immunoturbidimetric Albumin Method on Cobas c 501 Gives Higher
           Values Than the Abbott and Roche BCP Methods When Analyzing Patient Plasma
    • Authors: Johanna Helmersson-Karlqvist; Mats Flodin, Aleksandra Mandic Havelka, Xiao Yan Xu, Anders Larsson
      Pages: 677 - 681
      Abstract: BackgroundSerum/plasma albumin is an important and widely used laboratory marker and it is important that we measure albumin correctly without bias. We had indications that the immunoturbidimetric method on Cobas c 501 and the bromocresol purple (BCP) method on Architect 16000 differed, so we decided to study these methods more closely.MethodA total of 1,951 patient requests with albumin measured with both the Architect BCP and Cobas immunoturbidimetric methods were extracted from the laboratory system. A comparison with fresh plasma samples was also performed that included immunoturbidimetric and BCP methods on Cobas c 501 and analysis of the international protein calibrator ERM-DA470k/IFCC.ResultsThe median difference between the Abbott BCP and Roche immunoturbidimetric methods was 3.3 g/l and the Roche method overestimated ERM-DA470k/IFCC by 2.2 g/l. The Roche immunoturbidimetric method gave higher values than the Roche BCP method: y = 1.111x – 0.739, R² = 0.971.ConclusionThe Roche immunoturbidimetric albumin method gives clearly higher values than the Abbott and Roche BCP methods when analyzing fresh patient samples. The differences between the two methods were similar at normal and low albumin levels.
      PubDate: 2016-05-12T00:45:54.703886-05:
      DOI: 10.1002/jcla.21921
  • Analysis of Hemogram of Radiation Workers in Tangshan, China
    • Authors: Qing-Zeng Qian; Xiang-Ke Cao, Hai-Yan Liu, Fu-Hai Shen, Qian Wang, Jun-Wang Tong, Qing-Qiang Qian
      Pages: 682 - 688
      Abstract: ObjectivesThis study aimed to investigate changes in peripheral blood cells of radiation workers and explore the impact of long-term ionizing radiation (IR) on human peripheral hemogram.MethodsWith a cohort method, we selected 1,392 radiation workers (case group) and 1,430 non-health-ray-exposure history persons (control group) to detect and analyze their peripheral hemogram. FAITH3000 automatic biochemical analyzer was used for blood testing. Examination of peripheral hemogram includes the examination of white blood cells (WBCs), platelet (PLTs), red blood cells (RBCs), hemoglobin (Hb), lymphocytes (LYMs), and mononuclear cells (MOs). The data analysis was conducted with software SPSS19.0.ResultsAll the peripheral hemogram indicators (WBCs, RBCs, Hb, PLTs, LYMs, and MOs) in the case group, in accordance with the order of radiology diagnostic medical group, industrial inspection group, petroleum logging group, and radiotherapy medical group, showed a significant decreasing trend and were lower than those in the control group (all P < 0.05). Besides, with the increase of radiation seniority and accumulative radiation dose, all the peripheral hemogram indicators (WBCs, RBCs, Hb, PLTs, LYMs, and MOs) in the case group dramatically decreased and were lower than those in the control group (all P < 0.05). Seniority was in negative association with the expressions of WBCs, PLTs, RBCs, Hb, LYMs, and MOs and radiation dose with Hb, LYMs, and MOs (all P < 0.05).ConclusionLong-term IR has some effects on the health of radiation workers, thus protective measures should be further strengthened.
      PubDate: 2016-03-14T07:16:03.078733-05:
      DOI: 10.1002/jcla.21922
  • Influence of Phosphatidylethanolamine Concentration and Composition on the
           Detection of Antiphosphatidylethanolamine Antibodies by ELISA
    • Authors: Ke Ke; Zachariah I. Strango, Paul E. Harper, Ming Zhao
      Pages: 689 - 696
      Abstract: BackgroundAccumulating evidence supports a positive correlation between the presence of antiphosphatidylethanolamine (aPE) autoantibodies and clinical symptoms of antiphospholipid syndrome (APS). However, there is a lack of standardized ELISA-based method for detecting aPE. The current study was conducted to investigate the dependence of aPE ELISA on lipid concentration and composition of PE antigens.MethodsA range of ELISA conditions were examined by varying the concentrations of egg PE and by substituting egg PE with combinations of synthetic DOPE and DSPE. The physical properties of the synthetic PE species were also characterized.ResultsOur data indicated that there are different optimal PE concentrations for conducting ELISA assays for cofactor-dependent and cofactor-independent aPEs. In addition, using a two-component synthetic lipid system, we demonstrated aPE ELISA readouts can be modulated to approach the performance level of egg PE, which is currently the most commonly used PE antigen.ConclusionThese data raised the possibility of ultimately replacing natural PE antigens with a blend of defined synthetic lipid species, thus overcoming a known variable factor in aPE detection. The outcome of this study will help pave the way to developing a standardized aPE test.
      PubDate: 2016-04-07T23:57:56.813539-05:
      DOI: 10.1002/jcla.21923
  • Sensitivity of the Standard Chlamydia trachomatis Culture Method Is
           Improved After One Additional In Vitro Passage
    • Authors: Lili Shao†; Yuanli Guo†, Yong Jiang, Yuanjun Liu, Mei Wang, Cong You, Quanzhong Liu
      Pages: 697 - 701
      Abstract: BackgroundChlamydia trachomatis causes the most common bacterial sexually transmitted infection (STI) worldwide. Although highly sensitive nucleic acid amplification tests (NAATs) are used to routinely diagnose chlamydial infection, C. trachomatis isolation by cell culture is still preferred for legal cases and epidemiological studies because of its high specificity; however, the sensitivity of traditional two-passage diagnostic cultures is significantly lower than that of NAATs. Therefore, we sought to analyze if additional in vitro passaging of clinical samples would improve detection sensitivity of C. trachomatis.MethodsClinical swabs (n = 428) were collected from Tianjin Medical University General Hospital, grown in McCoy cells for up to five passages, and analyzed for the presence of inclusions by iodine staining. Results were confirmed by routine PCR-based methods.ResultsViable C. trachomatis organisms were detected in 91 (21.26%) swabs with the traditional two-passage protocol, which increased to 145 (33.88%) and 149 (34.81%) following three and four passages, respectively. Thus, the standard protocol yielded a false-negative rate of nearly 39%. Subsequent PCR-based diagnostics revealed a concordance rate of 80.98% between these two methods without any false negatives.ConclusionThe results of this study support the use of a three-passage Chlamydia culture procedure to increase the detection sensitivity of this method.
      PubDate: 2016-03-14T07:16:15.623295-05:
      DOI: 10.1002/jcla.21924
  • Anti-Mitotic Spindle Apparatus Antoantibodies: Prevalence and Disease
           Association in Chinese Population
    • Authors: Qian Xi; Yongkang Wu, Lixin Li, Bei Cai, Junlong Zhang, Bin Yang, Lanlan Wang
      Pages: 702 - 708
      Abstract: BackgroundMitotic spindle apparatus (MSA) antibodies are rare findings with undefined clinical significance in clinical research. We aimed at investigating the prevalence and clinical significance of anti-MSA antibodies in Chinese population.MethodsBetween 2008 and 2013, a total of 180,180 patients were studied for the presence of anti-MSA antibodies. The clinical details and laboratory data of anti-MSA-positive patients were retrospectively collected and analyzed.ResultsOf the 180,180 patients tested, 68,640 patients presented with positive antinuclear antibodies (ANAs, 38.10%), but only 32 patients with positive anti-MSA antibodies (0.018%). Diagnoses were established in 22 of 32 patients: 16 connective tissue diseases (CTDs), mainly Sjogren syndrome (SS, 5/16), rheumatoid arthritis (RA, 4/16), and systemic lupus erythematosus (SLE, 3/16), and 6 nonautoimmune conditions. The most frequent clinical symptoms of the anti-MSA-positive patients were arthralgia and eyes and mouth drying. Additionally, 70% of anti-MSA antibodies were not associated with other ANAs, however, when associated, the most frequent ANA was anti-SSA.ConclusionsAnti-MSA antibodies have a low prevalence and female gender predominance. Anti-MSA antibodies are primarily associated with CTDs, mainly SS, RA, and SLE. The presence of anti-MSA antibodies might be the unique serological markers of the CTDs, especially when anti-SSA, SSB, and dsDNA antibodies are negative, or the level of RF is low.
      PubDate: 2016-03-14T07:16:07.394684-05:
      DOI: 10.1002/jcla.21925
  • The Establishment of an HE4-CLIA Method and the Combined Analysis of HE4
           and CA125 in Ovarian Cancer
    • Authors: Qiong Zhang; Cong-Rong Wang, Juan-Ping Yu, Qiang Ma, Wei-Wen Xu
      Pages: 709 - 718
      Abstract: BackgroundThe human epididymal secretory protein 4 (HE4) is a novel, verified biomarker for the early diagnosis of ovarian cancer.MethodsMagnetic beads were coated with capture antibodies and were used with acridinium ester labeled detection antibodies in a sandwich-type immunoassay. The patient's HE4 serum levels were measured simultaneously with the chemiluminescence immunoassay (CLIA) kit we developed and electrochemiluminescence immunoassay (ECLIA) kit from Roche (Mannheim, Germany). CA125 was also detected by time-resolved fluoroimmunoassay. The diagnostic value was analyzed.ResultsThe assay demonstrated a linear range from 2.5 to 2,000 pmol/l, with an analytical sensitivity of 2.5 pmol/l. The reproducibility, recovery, and specificity of the immunoassay were demonstrated to be acceptable. Compared with the ECLIA kit from Roche in 124 serum samples (40 patients with ovarian cancer, 35 patients with benign gynecological diseases, and 49 health controls), there is a satisfied correlation coefficient of 0.875. The area under the receiver-operating curve (ROC-AUC) was 0.903 (95% CI was 0.839–0.966, P < 0.001) for HE4, 0.787 (95% CI was 0.694–0.879, P < 0.001) for CA125, and 0.914 (95% CI was 0.866–0.962, P < 0.001) for combined analysis of HE4 and CA125.ConclusionsA quantitative method (HE4-CLIA) for detecting HE4 in serum was successfully established. Preliminary clinical sample analysis showed HE4-CLIA has a certain clinical value in the screening and diagnosis of ovarian cancer. Moreover, in distinguishing benign from malignant ovarian lesions, HE4 has higher demonstrated accuracy than CA125.
      PubDate: 2016-03-17T02:42:08.431791-05:
      DOI: 10.1002/jcla.21926
  • A Simple, Rapid, and Highly Sensitive Electrochemical DNA Sensor for the
           Detection of α- and β-Thalassemia in China
    • Authors: Pei-Qi Chen; Qian-Ni Liang, Tao-Sheng Huang, Tian-Cai Liu, Ming Li
      Pages: 719 - 726
      Abstract: BackgroundBecause of the life-consuming treatment and severe consequences associated with thalassemia, it is more effective to prevent than cure thalassemia. Rapid and sensitive detection is critical for controlling thalassemia. In this study, we developed a rapid and accurate test to genotype nondeletional α- and β-thalassemia mutations by an electrochemical DNA sensor.MethodsScreen-printed electrodes were used as electrochemical transducers for the sensor, in which the capture probe DNA was attached to the golden surface of the working electrode via an S–Au covalent bond, which is highly suitable for immobilizing the biological element. In addition, two types of ferrocene with varying redox potentials for modified signal probe DNA were adopted. The hybridization signal is detected by alternating current voltammetry when the capture probe and signal probe hybridize with the target DNA.ResultsWith this technique, 12 types of nondeletional α- and β-thalassemia mutations were detected, which constitute more than 90% of all the nondeletional types of thalassemia mutation determinants found in China, including the CD142 (TAA>CAA) Constand spring, CD125 (CTG>CCG) Quonsze, CD122 (CAC>CAG) Weastmead, −28 (A>G), Cap+1 (A>C), initiation codon (ATG>AGG), CD17 (AAG>TAG), CD26 (GAG>AAG), CD31(-C), CD41-42 (-CTTT), CD71-72 (+A), and IVS-II-654 (C>T) mutations. Concordance levels were 100% within the 20 blood samples of homozygous wild-type individuals and 238 blood samples of heterozygous mutant individuals.ConclusionsThe electrochemical DNA sensor developed here can be applied for rapid genotyping of thalassemia or other clinical genotyping applications and is useful for early screening of thalassemia in high-risk groups by minimizing the time and investment cost.
      PubDate: 2016-05-26T07:56:17.500719-05:
      DOI: 10.1002/jcla.21927
  • The Association Between Circulating Levels of miRNA-181a and Pancreatic
           Beta Cells Dysfunction via SMAD7 in Type 1 Diabetic Children and
    • Authors: Enas Samir Nabih; Nevine Gamal Andrawes
      Pages: 727 - 731
      Abstract: BackgroundmiRNA-181a has been implicated in autoimmunity and apoptosis. Therefore, this study was conducted to explore its possible role in pancreatic beta-cells dysfunction.MethodsmiRNA-181a expression was evaluated by real-time PCR in serum of 40 type 1 diabetic children and adolescents and 40 age- and gender-matched healthy controls.ResultsmiRNA-181a expression was significantly higher in diabetic children and adolescents and it was negatively correlated to fasting C-peptide and SMAD7 levels.ConclusionmiRNA-181a appears to play a potential role in pancreatic beta-cells dysfunction via SMAD7.
      PubDate: 2016-02-18T21:01:23.854031-05:
      DOI: 10.1002/jcla.21928
  • Analytical Evaluation of Free Testosterone and Cortisol Immunoassays in
           Saliva as a Reliable Alternative to Serum in Sports Medicine
    • Authors: Giuseppe Lippi; Mariella Dipalo, Ruggero Buonocore, Cecilia Gnocchi, Rosalia Aloe, Roberto Delsignore
      Pages: 732 - 735
      Abstract: BackgroundThis study was aimed to investigate whether measurement of free testosterone and cortisol in saliva is a reliable alternative to their assessment in serum for monitoring physical fitness in professional athletes.MethodsWe studied 25 members of the soccer team Parma F.C., playing in Italian major football league. Blood and saliva samples were collected at fasting, before a regular training session. Cortisol, total and free testosterone, as well as the ratio between free testosterone and cortisol, were assessed in paired serum and saliva samples, and their results were compared.ResultsAn excellent correlation was found between serum and saliva cortisol (r = 0.751; P < 0.001). A significant correlation was also observed between free testosterone in serum and saliva (r = 0.590; P = 0.002), whereas no significant correlation was found between total testosterone in serum and saliva (r = 0.181; P = 0.387). A significant correlation was found for the free testosterone to cortisol ratio in serum and saliva (r = 0.43; P = 0.031). All athletes (25/25; 100%) declared that they would feel more comfortable to have saliva rather than blood serially collected.ConclusionsThe results of this study suggest that measurement of free testosterone and cortisol in saliva may be seen as a reliable alternative to their assessment in serum.
      PubDate: 2016-03-15T23:30:47.447167-05:
      DOI: 10.1002/jcla.21929
  • Serum Thymosin β4 Concentrations in Obstructive Sleep Apnea Syndrome
    • Authors: Yongquan Liu; Meijuan Liu, Youkui Shi, Yuan Liu
      Pages: 736 - 740
      Abstract: ObjectiveInflammation is a potential mechanism of obstructive sleep apnea syndrome (OSAS). Thymosin β4, a member of thymic protein family, exhibits an anti-inflammatory effect. We determine to investigate whether serum thymosin β4 concentrations is correlated with the occurrence and disease severity of OSAS.MethodsSerum thymosin β4 concentrations were examined in a cross-sectional population including 158 patients with OSAS and 94 healthy subjects.ResultsElevated serum thymosin β4 concentrations were found in OSAS patients than the controls. Multivariable logistic regression analysis indicated a significant association between serum thymosin β4 concentrations and OSAS development. Severe OSAS patients showed increased serum thymosin β4 concentrations compared with mild and moderate patients. Spearman correlation analysis suggested that serum thymosin β4 concentrations were correlated with the severity of OSAS. Simple linear regression analyses showed that serum thymosin β4 in OSAS patients was correlated with homeostasis model assessment of insulin resistance, apnea hypopnea index, disease severity, and osteoarthritis development. Then multiple stepwise regression analysis showed that only disease severity remained to be associated with serum thymosin β4.ConclusionsSerum thymosin β4 concentrations were correlated with the occurrence and severity of OSAS.
      PubDate: 2016-04-18T02:17:27.239038-05:
      DOI: 10.1002/jcla.21930
  • Seasonal Variation of Blood Calcium Levels in Children Aged 1–10
    • Authors: Meichun Zhang; Rongrong Zhai, Jie Liu, Hui Guang, Benzhong Li, Songtao Zhang
      Pages: 741 - 744
      Abstract: BackgroundIn this study, the associations of seasons with blood calcium levels in children aged 1–10 have not been evaluated.MethodsIn 2012–2014, whole blood samples were collected from 2,562 children and calcium concentrations were determined by flame atomic absorption spectrometry. The associations of seasons with calcium levels were analyzed by multivariable regression.ResultsThe mean value of calcium concentrations was 1.61 ± 0.13 mmol/l and the overall deficiency was 29.3%. Overall, compared to those in winter, children in spring and summer had significant lower calcium concentrations that decreased by 1.2% (β = −0.012; 95% CI: −0.021, −0.002) and 1.4% (β = −0.014; 95% CI: −0.023, −0.005), respectively; and corresponding higher calcium deficiencies than those in spring, summer, and autumn with odds ratios (OR) were 1.93 (95% CI: 1.39, 2.66), 1.65 (95% CI: 1.21, 2.24), and 1.57 (95% CI: 1.14, 2.15), respectively. Moreover, this seasonality was more significant in girls in whom calcium concentration in summer decreased by 1.9% (β = −0.019; 95% CI: −0.036, −0.003) and OR for calcium deficiencies in summer was 2.46 (1.38–4.41), compared to the girls in winter.ConclusionsThe seasons have significant association with blood calcium levels, especially in girls. However, the impact of this seasonality on children's health is still unknown.
      PubDate: 2016-04-07T23:57:27.745578-05:
      DOI: 10.1002/jcla.21931
  • Conventional Morphology Versus PCR Sequencing, rep-PCR, and MALDI-TOF-MS
           for Identification of Clinical Aspergillus Isolates Collected Over a
           2-Year Period in a University Hospital at Kayseri, Turkey
    • Authors: Altay Atalay; Ayse Nedret Koc, Ahmet Suel, Hafize Sav, Gonca Demir, Ferhan Elmali, Nuri Cakir, Seyedmojtaba Seyedmousavi
      Pages: 745 - 750
      Abstract: BackgroundAspergillus species cause a wide range of diseases in humans, including allergies, localized infections, or fatal disseminated diseases. Rapid detection and identification of Aspergillus spp. facilitate effective patient management. In the current study we compared conventional morphological methods with PCR sequencing, rep-PCR, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for the identification of Aspergillus strains.Materials and MethodsA total of 24 consecutive clinical isolates of Aspergillus were collected during 2012–2014. Conventional morphology and rep-PCR were performed in our Mycology Laboratory. The identification, evaluation, and reporting of strains using MALDI-TOF-MS were performed by BioMérieux Diagnostic, Inc. in Istanbul. DNA sequence analysis of the clinical isolates was performed by the BMLabosis laboratory in Ankara.ResultsSamples consisted of 18 (75%) lower respiratory tract specimens, 3 otomycosis (12.5%) ear tissues, 1 sample from keratitis, and 1 sample from a cutaneous wound. According to DNA sequence analysis, 12 (50%) specimens were identified as A. fumigatus, 8 (33.3%) as A. flavus, 3 (12.5%) as A. niger, and 1 (4.2%) as A. terreus. Statistically, there was good agreement between the conventional morphology and rep-PCR and MALDI-TOF methods; kappa values were κ = 0.869, 0.871, and 0.916, respectively (P < 0.001).ConclusionThe good level of agreement between the methods included in the present study and sequence method could be due to the identification of Aspergillus strains that were commonly encountered. Therefore, it was concluded that studies conducted with a higher number of isolates, which include other Aspergillus strains, are required.
      PubDate: 2016-03-01T04:52:24.905598-05:
      DOI: 10.1002/jcla.21932
  • Resazurin Microtiter Assay for Clarithromycin Susceptibility Testing of
           Clinical Isolates of Mycobacterium abscessus Group
    • Authors: Natalia Fernandes Carvalho; Daisy Nakamura Sato, Fernando R. Pavan, Lucilaine Ferrazoli, Erica Chimara
      Pages: 751 - 755
      Abstract: BackgroundMycobacterium abscessus group has heterogeneous susceptibility pattern among species. The species is most common cause of nosocomial infections. Macrolides minimum Inhibitory concentration (MIC) determination is essential for the treatment.MethodsThirty-six strains were randomly selected for performing Resazurin Microtiter Assay (REMA) for clarithromycin testing in comparison to MIC test according to Clinical and Laboratory Standards Institute (2011) recommendation. REMA has been used for detection of drug resistance in M. tuberculosis. Extended incubation was performed to detect induced resistance.ResultsThirty microliters of resazurin (0.01%) was added after visually taking MIC reading. Resistance was observed in 11.1% of M. bolletti and 4.8% of M. abscessus strains; and induced resistance was detected in 77.8% and 95.2% of M. bolletti and M. abscessus strains, respectively. All strains of M. massiliense were susceptible. The samples presented same MIC value both by visual reading and through resazurin.ConclusionThe present study showed 100% concordance between both readings, with REMA providing easier to read and report results benefit. This change in reading can also reflect on the MIC determination and report, improving the test.
      PubDate: 2016-05-12T00:45:34.372766-05:
      DOI: 10.1002/jcla.21933
  • Comparison of a New and Rapid Method: Brucella Coombs Gel Test With Other
           Diagnostic Tests
    • Authors: Fatma Kalem; Ayşe Gül Ergün, Süleyman Durmaz, Metin Doğan, Ömür Ertuğrul, Seval Gündem
      Pages: 756 - 759
      Abstract: BackgroundThe aim of this study was to detect reliability of Brucella Coombs gel test (BCGT) by comparing with with ELISA (IgG + IgM), Standard agglutination test, and Brucella immunocapture agglutination methods in serological diagnosis of brucellosis.MethodsBrucella Coombs gel test (BCGT), Brucella ELISA (IgG + IgM), Standard agglutination test, and Brucella immunocapture agglutination tests of 78 patients with presumptive diagnosis of brucellosis which were sent to Microbiology Laboratory of Konya Numune Hospital from various regions of Konya were studied.ResultsResults: Of 78 patients with ELISA IgG and IgM, STA, BICA and BCGT; 26, 21, 10, 12 and 12 were positive. When compared with BICA, the sensitivity and specifity of BCGT were 100% and 100%, respectively.ConclusionAccording to results BCGT can be used as a diagnostic test in routine laboratories after more comprehensive studies in control groups and patients.
      PubDate: 2016-03-17T02:41:43.142447-05:
      DOI: 10.1002/jcla.21934
  • Hemoculture and Direct Sputum Detection of mecA-Mediated
           Methicillin-Resistant Staphylococcus aureus by Loop-Mediated Isothermal
           Amplification in Combination With a Lateral-Flow Dipstick
    • Authors: Kawin Nawattanapaiboon; Photchanathorn Prombun, Pitak Santanirand, Apirom Vongsakulyanon, Toemsak Srikhirin, Boonsong Sutapun, Wansika Kiatpathomchai
      Pages: 760 - 767
      Abstract: This study reports loop-mediated isothermal amplification (LAMP) for rapid detection of methicillin-resistant Staphylococcus aureus from direct clinical specimens. Four primers including outer and inner primers were specifically designed on the two target sequences—femB to identify S. aureus and mecA to identify antibiotic-resistant gene. Reference strains including various species of gram-positive/gram-negative isolates were used to evaluate and optimize LAMP assays. The optimum LAMP condition was found at 63°C within 70 min assay time (include hybridization with FITC probe for 5 min and further 5 min for reading the results on the lateral flow dipstick). The detection limits of LAMP for mecA was 10 pg of total DNA or 100 CFU/ml. The LAMP assays were applied to a total of 155 samples of direct DNA extraction from sputum and hemoculture bottles. The sensitivity of LAMP for mecA detection in sputum and hemoculture bottles was 93.3% (28/30) and 100% (52/52), respectively. In conclusion, LAMP assay is an alternative technique for rapid detection of MRSA infection with a technical simplicity and cost-effective method in a routine diagnostic laboratory.
      PubDate: 2016-03-17T02:41:28.906561-05:
      DOI: 10.1002/jcla.21935
  • Relationship Between the Serum Total Bilirubin and Inflammation in
           Patients With Psoriasis Vulgaris
    • Authors: Zhen-Xing Zhou; Jian-Kui Chen, Yan-Ying Hong, Ru Zhou, Dong-Mei Zhou, Li-Yun Sun, Wen-Li Qin, Tian-Cheng Wang
      Pages: 768 - 775
      Abstract: BackgroundPsoriasis is a chronic and recurrent inflammatory skin disease. Previous studies have shown that bilirubin has anti-inflammation and antioxidant effects. However, the various roles of bilirubin in psoriasis patients are still unclear.ObjectiveTo investigate the serum total bilirubin (TB) level in the individuals with psoriasis vulgaris and further evaluate the relationship between serum TB concentration and C-reactive protein (CRP) to clarify the effect of bilirubin on inflammation.MethodsA total of 214 patients with psoriasis vulgaris and 165 age- and gender-matched healthy control subjects were recruited. The peripheral leukocyte count (white blood cell, WBC) and differential, serum biochemical and immunologic indexes including serum TB, immunoglobulin (Ig) G, IgA, IgM, complement C3 and C4, as well as serum CRP concentrations were measured.ResultsResults showed that the serum TB level decreased significantly and peripheral WBC, neutrophil, and serum CRP concentrations increased significantly in patients with psoriasis vulgaris. Meanwhile, the serum CRP was negatively correlated with serum TB levels but positively correlated with peripheral WBC and the Psoriasis Area and Severity Index (PASI). Logistic regression analysis showed that the serum TB was a protective factor for psoriasis vulgaris.ConclusionThe present study suggests that lower serum TB is associated with the enhancement of the inflammatory response in psoriasis vulgaris. Therefore, lower serum TB has a prognostic significance for worsening psoriasis vulgaris. Bilirubin may play a crucial role in inflammation by contributing to the inhibition of the inflammatory response.
      PubDate: 2016-04-07T23:57:10.38841-05:0
      DOI: 10.1002/jcla.21936
  • Neutrophil and Eosinophil Granule Proteins as Potential Biomarkers of
           Assessing Disease Activity and Severity in Patients With Ulcerative
    • Authors: Zhiyan Li; Yan Long, Mingjian Bai, Junxia Li, Zhenru Feng
      Pages: 776 - 778
      Abstract: BackgroundColonoscopy can assess disease activity and severity of ulcerative colitis (UC) accurately, but it is invasive and costly. Role of noninvasive biomarkers of intestinal inflammation in evaluation of patients with UC is not well understood. In this study, we assessed fecal eosinophil cationic protein (FECP), fecal myeloperoxidase (FMPO), and fecal calprotectin (FC) as surrogate markers of disease activity and severity in patients with UC, and then evaluated effect of the combination of these markers.MethodsSixty-three UC patients and 59 cases of age-matched controls were investigated. All patients underwent clinical, endoscopic, and histological assessment for disease activity and severity. Fecal samples were analyzed for FECP, FC, and FMPO.ResultsAll three fecal biomarkers were elevated in patients compared with controls (P = 0.000). Significant differences were found between inactive UC and controls (P = 0.000). Cases with severe UC had significantly higher FECP levels than those with mild UC (p < 0.05), but there were no significant differences in FC and FMPO levels among disease severity groups. All three biomarkers showed positive correlation with Ulcerative Colitis Activity Index (UCAI). The areas under the ROC curve of FECP, FC, and FMPO were 0.939, 0.783, and 0.785, respectively. Sensitivity and specificity of fecal biomarkers in assessing disease activity were FECP—88.46%, 89.47%; FC—80.77%, 68.42%; and FMPO—84.62%, 63.16%.ConclusionsAll three fecal biomarkers could be used as surrogate markers for assessing disease activity of UC, and FECP provided superior discrimination than FMPO and FC. Moreover, FECP could distinguish between mild disease and severe disease group.
      PubDate: 2016-04-13T20:47:57.505885-05:
      DOI: 10.1002/jcla.21937
  • Detection of Apoptotic Lymphocytes Through Sysmex XN-1000 As a Diagnostic
           Marker for Mononucleosis Syndrome
    • Authors: Silvia Sale; Addolorata Emanuela Carone, Maurizio Fumi, Ylenia Pancione, Vincenzo Rocco
      Pages: 779 - 793
      Abstract: BackgroundThe infectious mononucleosis (IM) includes clue elements, apoptotic and atypical lymphocytes. In IM, the evaluation of dot plot provided by Sysmex XN-1000 analyzer revealed a stretched lymphocytic cluster, white cell differential channel (WDF), on cytogram.MethodsIn this study, we analyzed 698 samples that include 39 IM, 76 chronic lymphoproliferative disorders, 25 nonclonal lymphocytosis, and 40 healthy donors. Five hundred eighteen samples with other diseases or interference were evaluated. The algorithm was validated on 40,000 files that were received from internal database of Sysmex-Dasit.ResultsThe analysis of flow cytometry standard (FCS) files in WDF channel and presumed apoptotic lymphocytes counts on side scatter/forward scatter (SSC/FSC) and SSC/SFL (where SFL is side fluorescence) dot plot revealed excellent correlation among apoptotic cells on peripheral blood smear (R2 = 0.79 and 0.75). There was a variation of positional parameters in lymphocyte clusters WX, WY, and WZ. If WX-SSC > 500 and WY-SFL > 1,000 and WZ-FSC > 700, specificity equals to 99% and sensitivity equals to 100%. If nucleated red blood cell (NRBC)
      PubDate: 2016-04-19T22:31:21.427523-05:
      DOI: 10.1002/jcla.21938
  • Issue Information - TOC
    • First page: 1527
      Abstract: No abstract is available for this article.
      PubDate: 2016-10-12T22:33:10.956106-05:
      DOI: 10.1002/jat.3246
  • Revision of the affinity constant for perchlorate binding to the
           sodium‐iodide symporter based on in vitro and human in vivo data
    • Authors: Paul M. Schlosser
      First page: 1531
      Abstract: A series of previously published physiologically based pharmacokinetic (PBPK) models describe the effect of perchlorate on iodide uptake by the thyroid, with the mechanism being competitive inhibition of iodide transport by the sodium‐iodide symporter (NIS). Hence a key parameter of these models is the affinity of perchlorate for the NIS, characterized as the Michaelis–Menten kinetic constant, Km. However, when model predictions were compared to published results of a human study measuring radio‐iodide uptake (RAIU) inhibition after controlled perchlorate exposures, it was found to only fit the lowest exposure level and underpredicted RAIU inhibition at higher levels. Published in vitro data, in which perchlorate‐induced inhibition of iodide uptake via the NIS was measured, were re‐analyzed. Km for binding of perchlorate to the NIS originally derived from these data, 1.5 μm, had been obtained using Lineweaver–Burk plots, which allow for linear regression but invert the signal–noise of the data. Re‐fitting these data by non‐linear regression of the non‐inverted data yielded a 60% lower value for the Km, 0.59 μm. Substituting this value into the PBPK model for an average adult human significantly improved model agreement with the human RAIU data for exposures
      PubDate: 2016-05-13T08:40:22.50649-05:0
      DOI: 10.1002/jat.3337
  • Integrated approach to testing and assessment for predicting rodent
           genotoxic carcinogenicity
    • Authors: Petko I. Petkov; Terry W. Schultz, E. Maria Donner, Masamitsu Honma, Takeshi Morita, Shuichi Hamada, Akihiro Wakata, Masayuki Mishima, Jiro Maniwa, Milen Todorov, Elena Kaloyanova, Stefan Kotov, Ovanes G. Mekenyan
      First page: 1536
      Abstract: We investigated the performance of an integrated approach to testing and assessment (IATA), designed to cover different genotoxic mechanisms causing cancer and to replicate measured carcinogenicity data included in a new consolidated database. Genotoxic carcinogenicity was predicted based on positive results from at least two genotoxicity tests: one in vitro and one in vivo (which were associated with mutagenicity categories according to the Globally Harmonized System classification). Substances belonging to double positives mutagenicity categories were assigned to be genotoxic carcinogens. In turn, substances that were positive only in a single mutagenicity test were assigned to be mutagens. Chemicals not classified by the selected genotoxicity endpoints were assigned to be negative genotoxic carcinogens and subsequently evaluated for their capability to elicit non‐genotoxic carcinogenicity. However, non‐genotoxic carcinogenicity mechanisms were not currently included in the developed IATA. The IATA is docked to the OECD Toolbox and uses measured data for different genotoxicity endpoints when available. Alternatively, the system automatically provides predictions by SAR genotoxicity models using the OASIS Tissue Metabolism Simulator platform. When the developed IATA was tested against the consolidated database, its performance was found to be high, with sensitivity of 74% and specificity of 83%, when measured carcinogenicity data were used along with predictions falling within the models' applicability domains. Performance of the IATA would be slightly changed to a sensitivity of 80% and specificity of 72% when the evaluation by non‐genotoxic carcinogenicity mechanisms was taken into account. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-05-25T22:50:32.162628-05:
      DOI: 10.1002/jat.3338
  • Danio rerio ABC transporter genes abcb3 and abcb7 play a protecting role
           against metal contamination
    • Authors: Adélaïde Lerebours; Van Vinh To, Jean-Paul Bourdineaud
      First page: 1551
      Abstract: ATP-binding cassette (ABC) proteins are efflux transporters and some of them are involved in xenobiotic detoxification. The involvement of four zebrafish ABC transporters in cadmium, zinc and mercury detoxification was characterized in a metal hypersensitive mutant of Escherichia coli. The E. coli tolC mutant expressing ABCB3 or ABCB7 transporters exhibited higher survival ratios and lower metal accumulation under a metal exposure condition than the controls. For instance, in the presence of 8 and 10 μM of HgCl2, the survival ratios of bacteria expressing ABCB3 were four and six-times higher than the control whereas the mercury concentrations were 2.5 and 2-times lower than in the control. This work provides new data on the function of zebrafish ABCB3 and ABCB7 transporters and highlights their significance in metal detoxification. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-03-28T06:05:52.841222-05:
      DOI: 10.1002/jat.3313
  • Different mechanisms of action of 2, 2’, 4, 4’-tetrabromodiphenyl
           ether (BDE-47) and its metabolites (5-OH-BDE-47 and 6-OH-BDE-47) on cell
           proliferation in OVCAR-3 ovarian cancer cells and MCF-7 breast cancer
    • Authors: Anna Karpeta; Anna Maniecka, Ewa Łucja Gregoraszczuk
      First page: 1558
      Abstract: Data concerning the possible action of polybrominated diphenyl ethers (PBDEs) in hormone-dependent cancer are scarce. Some data showed that PBDEs may directly affect breast cancer cells formation and only one research showed increased proliferation of the OVCAR-3 cells, but the results are ambiguous and the mechanisms are not clear. There is growing evidence that not only parent compounds but also its metabolites may be involved in cancer development. The present study was, therefore, designed to determine the effect of BDE-47 and its metabolites (2.5 to 50 ng ml–1) on proliferation (BrdU), cell-cycle genes (real-time PCR) and protein expression (Western blot), protein expression of oestrogen receptors (α β), extracellular signal-regulated kinases 1 and 2 (ERK1/2) and protein kinase Cα (PKCα) in OVCAR-3 ovarian and MCF-7 breast cancer cells. In OVCAR-3 cells, the parent compound stimulated cell proliferation by activating CDK1, CDK7, E2F1 and E2F2. Independent of time of exposure, BDE-47 had no effect on ERα and ERβ protein expression and ERK1/2 and PKCα phosphorylation. Metabolites had no effect on cell proliferation but increased both ERs protein expression and ERK1/2 and PKCα phosphorylation. In MCF-7 cells, the parent compound displayed no effect on cell proliferation but decreased ERα and increased ERβ protein expression with concomitant induction of PKCα phosphorylation. Both metabolites increased MCF-7 cell proliferation, ERK1/2 and PKCα phosphorylation and decreased ERα and ERβ protein expression.We suggest that studies concerning PBDEs with fewer bromine atoms should be continued to understand environmental links to different hormone-dependent cancers. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-03-14T07:48:03.50754-05:0
      DOI: 10.1002/jat.3316
  • Accounting for data variability, a key factor in in vivo/in vitro
           relationships: application to the skin sensitization potency (in vivo LLNA
           versus in vitro DPRA) example
    • Authors: S. Dimitrov; A. Detroyer, C. Piroird, C. Gomes, J. Eilstein, T. Pauloin, C. Kuseva, H. Ivanova, I. Popova, Y. Karakolev, S. Ringeissen, O. Mekenyan
      First page: 1568
      Abstract: When searching for alternative methods to animal testing, confidently rescaling an in vitro result to the corresponding in vivo classification is still a challenging problem. Although one of the most important factors affecting good correlation is sample characteristics, they are very rarely integrated into correlation studies. Usually, in these studies, it is implicitly assumed that both compared values are error-free numbers, which they are not. In this work, we propose a general methodology to analyze and integrate data variability and thus confidence estimation when rescaling from one test to another. The methodology is demonstrated through the case study of rescaling the in vitro Direct Peptide Reactivity Assay (DPRA) reactivity to the in vivo Local Lymph Node Assay (LLNA) skin sensitization potency classifications. In a first step, a comprehensive statistical analysis evaluating the reliability and variability of LLNA and DPRA as such was done. These results allowed us to link the concept of gray zones and confidence probability, which in turn represents a new perspective for a more precise knowledge of the classification of chemicals within their in vivo OR in vitro test. Next, the novelty and practical value of our methodology introducing variability into the threshold optimization between the in vitro AND in vivo test resides in the fact that it attributes a confidence probability to the predicted classification. The methodology, classification and screening approach presented in this study are not restricted to skin sensitization only. They could be helpful also for fate, toxicity and health hazard assessment where plenty of in vitro and in chemico assays and/or QSARs models are available. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-04-15T03:45:36.191466-05:
      DOI: 10.1002/jat.3318
  • Pyrazinamide induced hepatic injury in rats through inhibiting the
           PPARα pathway
    • Authors: Yun Zhang; Hongli Guo, Hozeifa M. Hassan, Ping-ping Ding, Yijing Su, Yuming Song, Tao Wang, Lixin Sun, Luyong Zhang, Zhenzhou Jiang
      First page: 1579
      Abstract: Pyrazinamide (PZA) causes serious hepatotoxicity, but little is known about the exact mechanism by which PZA induced liver injury. The peroxisome proliferator-activated receptors alpha (PPARα) is highly expressed in the liver and modulates the intracellular lipidmetabolism. So far, the role of PPARα in the hepatotoxicity of PZA is unknown. In the present study, we described the hepatotoxic effects of PZA and the role of PPARα and its target genes in the downstream pathway including L-Fabp, Lpl, Cpt-1b, Acaa1, Apo-A1 and Me1 in this process. We found PZA induced the liver lipid metabolism disorder and PPARα expressionwas down-regulated which had a significant inverse correlation with liver injury degree. These changeswere ameliorated by fenofibrate, the co-treatment that acts as a PPARα agonist. In contrast, short-termstarvation significantly aggravated the severity of PZA-induced liver injury. In conclusion, this study demonstrated the critical role played by PPARα in PZA-induced hepatotoxicity and provided a better understanding of the molecular mechanisms underlying PZA-induced liver injury. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-04-12T21:16:46.631941-05:
      DOI: 10.1002/jat.3319
  • Perfluorooctane sulfonate (PFOS) impairs the proliferation of C17.2 neural
           stem cells via the downregulation of GSK-3β/β-catenin signaling
    • Authors: Xuan Dong; Jianbin Yang, Xiaoke Nie, Jing Xiao, Shengyang Jiang
      First page: 1591
      Abstract: The neurotoxic effects of perfluorooctane sulfonate (PFOS) have attracted significant research attention in recent years. In the present study, we investigated the impact of PFOS exposure on the physiology of neural stem cells (NSCs) in vitro. We showed that PFOS exposure markedly attenuated the proliferation of C17.2 neural stem cells in both dose- and time-dependent manners. Additionally, we found that PFOS decreased Ser9 phosphorylation of glycogen synthase kinase-3β (pSer9-GSK-3β), leading to the activation of GSK-3β and resultant downregulation of cellular β-catenin. Furthermore, blockage of GSK-3β with lithium chloride significantly attenuated both the PFOS-induced downregulation of GSK-3β/β-catenin and the proliferative impairment of C17.2 cells. Notably, the expression of various downstream targets was altered accordingly, such as c-myc, cyclin D1 and survivin. In conclusion, the present study demonstrated that PFOS decreased the proliferation of C17.2 cells via the negative modulation of the GSK-3β/β-catenin pathway. We present the potential mechanisms underlying the PFOS-induced toxic effects on NSCs to provide novel insights into the neurotoxic mechanism of PFOS. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-03-28T06:15:37.369126-05:
      DOI: 10.1002/jat.3320
  • Accessing the molecular interactions of phthalates and their primary
           metabolites with the human pregnane X receptor using in silico profiling
    • Authors: M. K. Sarath Josh; S. Pradeep, Aparna K. Balan, M. N. Sreejith, Sailas Benjamin
      First page: 1599
      Abstract: Phthalates are known to cause endocrine disruption in humans and animals. Being lipophilic xenobiotic chemicals, phthalates from the surrounding environments can easily be absorbed into the biological system, thereby causing various health dysfunctions. This molecular docking study evaluates a variety of molecular interactions of 12 commonly used diphthalates and respective monophthalates onto the ligand binding domain (LBD) of the human pregnane X receptor (hPXR), a xenosensor, which would be beneficial for further in vitro and in vivo studies on hazardous phthalates. Out of 12 diphthalates and their monophthalates tested, diisodecyl phthalate (–9.16 kcal mol–1) showed more affinity toward hPXR whereas diisononyl phthalate (–8.77) and di(2-ethyhexyl)phthalate (–8.56), the predominant plasticizers found in a variety of plastics and allied products, showed comparable binding scores with that of the control ligands such as hyperforine (–9.99) and dexamethasone (–7.36). In addition to the above diphthalates, some of their monophthalates (monoisodecyl phthalate, mono-2-etheylhexyl phthalate, etc.) also established similar interactions with certain crucial amino acids in the LBD, which led to higher G scores. In fact, bisphenol A, a well-studied and proven endocrine disruptor, showed lesser G scores (–6.69) than certain phthalates. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-04-12T21:21:30.335434-05:
      DOI: 10.1002/jat.3321
  • The neurotoxicity of DE-71: effects on neural development and impairment
           of serotonergic signaling in zebrafish larvae
    • Authors: Xianfeng Wang; Lihua Yang, Qiangwei Wang, Yongyong Guo, Na Li, Mei Ma, Bingsheng Zhou
      First page: 1605
      Abstract: The underlying mechanism of polybrominated diphenyl ether (PBDE)-induced neurotoxicity is still a major concern due to its ubiquitous nature and persistence. Here, zebrafish embryos (2 h postfertilization, hpf) were exposed to different concentrations of the commercial PBDE mixture DE-71 (0–100 µg l–1) until 120 hpf, and the impact on neural development and serotonergic system was investigated. The in vivo results revealed significantly reduced transcription of genes involved in neurogenesis (fgf8, shha, wnt1), and contents of proteins in neuronal morphogenesis (myelin basic protein, synapsin IIa), suggesting an impairment of neural development in zebrafish embryos. Further results demonstrated a reduction of 5-hydroxytryptamine neuron and a dose-dependent decrease of whole-body serotonin levels, as well as the transcription of genes involved in serotonergic synthesis (tph1, tph2, trhr) and neurotransmission (serta/b, htr1aa/b). In addition, we predicted possible targets of PBDEs by molecular docking, and the results indicated that PBDE congeners showed high binding affinities with fibroblast growth factor 8 other than SHH and HTR1B. Taken together, this study demonstrated that PBDE exposure during embryogenesis could damage neural development and cause impairment of the serotonergic system as secondary effects in the zebrafish larvae. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-03-22T04:45:38.166693-05:
      DOI: 10.1002/jat.3322
  • MicroRNA profiles in a monkey testicular injury model induced by
           testicular hyperthermia
    • Authors: Ken Sakurai; Kei Mikamoto, Makoto Shirai, Takuma Iguchi, Kazumi Ito, Wataru Takasaki, Kazuhiko Mori
      First page: 1614
      Abstract: To characterize microRNAs (miRNAs) involved in testicular toxicity in cynomolgus monkeys, miRNA profiles were investigated using next-generation sequencing (NGS), microarray and reverse transcription-quantitative real-time-PCR (RT-qPCR) methods. First, to identify organ-specific miRNAs, we compared the expression levels of miRNAs in the testes to those in representative organs (liver, heart, kidney, lung, spleen and small intestine) obtained from naïve mature male and female monkeys (n = 2/sex) using NGS analysis. Consequently, miR-34c-5p, miR-202-5p, miR-449a and miR-508-3p were identified to be testicular-specific miRNAs in cynomolgus monkeys. Next, we investigated miRNA profiles after testicular–hyperthermia (TH) treatment to determine which miRNAs are involved in testicular injury. In this experiment, mature male monkeys were divided into groups with or without TH-treatment (n = 3/group) by immersion of the testes in a water bath at 43 °C for 30 min for 5 consecutive days. As a result, TH treatment induced testicular injury in all animals, which was characterized by decreased numbers of spermatocytes and spermatids. In a microarray analysis of the testis, 11 up-regulated (>2.0 fold) and 13 down-regulated (
      PubDate: 2016-04-12T21:45:55.640689-05:
      DOI: 10.1002/jat.3326
  • A 28-year observational study of urinary cadmium and β2-microglobulin
           concentrations in inhabitants in cadmium-polluted areas in Japan
    • Authors: Hoang Duc Phuc; Teruhiko Kido, Ho Dung Manh, Le Thai Anh, Nguyen Thi Phuong Oanh, Rie Okamoto, Akie Ichimori, Kazuhiro Nogawa, Yasushi Suwazono, Hideaki Nakagawa
      First page: 1622
      Abstract: The biological half-life of cadmium (Cd) is as long as 10–30 years. Exposure to this element induces renal tubular dysfunction, which is considered irreversible. β2-microglobulin (β2-MG) is a low-molecular-weight protein, and urinary β2-MG is one of the most useful and critical indicators for the early detection of renal tubular dysfunction. However, very little research has been published concerning the long-term observation of Cd-induced adverse health effects. As such, this follow-up study was conducted for 28 years to clarify the relationship between the concentration of Cd and β2-MG in the urine of 28 inhabitants (14 male and 14 female) living in the Kakehashi River basin, Ishikawa prefecture (Japan), previously one of the most highly Cd-polluted regions in this country. All subjects were over 60 years old in 2014 and participated in all six health examinations conducted over 28 years (1986–2014). Urine was collected at the appropriate time and kept frozen to analyze urinary Cd and β2-MG concentrations. The urinary Cd concentration was found to decrease by nearly half between 1986 and 2008 in both male and female subjects, whereas it increased significantly from 2008 to 2014 in males. In contrast, urinary β2-MG concentrations tended to increase over the 28-year study period in both sexes. Urinary Cd and β2-MG concentrations in females were significantly higher than those in males in this Cd-polluted region. Age is more strongly associated with urinary β2-MG concentration than recent Cd body burden. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-04-15T02:36:12.90087-05:0
      DOI: 10.1002/jat.3327
  • Development of the Larval Amphibian Growth and Development Assay: effects
           of chronic 4‐tert‐octylphenol or 17β‐trenbolone exposure in Xenopus
           laevis from embryo to juvenile
    • Authors: Jonathan T. Haselman; Patricia A. Kosian, Joseph J. Korte, Allen W. Olmstead, Taisen Iguchi, Rodney D. Johnson, Sigmund J. Degitz
      First page: 1639
      Abstract: The Larval Amphibian Growth and Development Assay (LAGDA) is a globally harmonized test guideline developed by the U.S. Environmental Protection Agency in collaboration with Japan's Ministry of the Environment. The LAGDA was designed to evaluate apical effects of chronic chemical exposure on growth, thyroid‐mediated amphibian metamorphosis and reproductive development. During the validation phase, two well‐characterized endocrine‐disrupting chemicals were tested to evaluate the performance of the initial assay design: xenoestrogen 4‐tert‐octylphenol (tOP) and xenoandrogen 17β‐trenbolone (TB). Xenopus laevis embryos were exposed, in flow‐through conditions, to tOP (nominal concentrations: 0.0, 6.25, 12.5, 25 and 50 µg l–1) or TB (nominal concentrations: 0.0, 12.5, 25, 50 and 100 ng l–1) until 8 weeks post‐metamorphosis, at which time growth measurements were taken, and histopathology assessments were made of the gonads, reproductive ducts, liver and kidneys. There were no effects on growth in either study and no signs of overt toxicity, sex reversal or gonad dysgenesis. Exposure to tOP caused a treatment‐related decrease in circulating thyroxine and an increase in thyroid follicular cell hypertrophy and hyperplasia (25 and 50 µg l–1) during metamorphosis. Müllerian duct development was affected after exposure to both chemicals; tOP exposure caused dose‐dependent maturation of oviducts in both male and female frogs, whereas TB exposure caused accelerated Müllerian duct regression in males and complete regression in >50% of the females in the 100 ng l–1 treatment. Based on these results, the LAGDA performed adequately to evaluate apical effects of chronic exposure to two endocrine‐active compounds and is the first standardized amphibian multiple life stage toxicity test to date. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.
      PubDate: 2016-05-03T07:51:03.166644-05:
      DOI: 10.1002/jat.3330
  • Development of the Larval Amphibian Growth and Development Assay: Effects
           of benzophenone‐2 exposure in Xenopus laevis from embryo to juvenile
    • Authors: Jonathan T. Haselman; Maki Sakurai, Naoko Watanabe, Yasushi Goto, Yuta Onishi, Yuki Ito, Yu Onoda, Patricia A. Kosian, Joseph J. Korte, Rodney D. Johnson, Taisen Iguchi, Sigmund J. Degitz
      First page: 1651
      Abstract: The Larval Amphibian Growth and Development Assay (LAGDA) is a globally harmonized chemical testing guideline developed by the U.S. Environmental Protection Agency in collaboration with Japan's Ministry of Environment to support risk assessment. The assay is employed as a higher tiered approach to evaluate effects of chronic chemical exposure throughout multiple life stages in a model amphibian species, Xenopus laevis. To evaluate the utility of the initial LAGDA design, the assay was performed using a mixed mode of action endocrine disrupting chemical, benzophenone‐2 (BP‐2). X. laevis embryos were exposed in flow‐through conditions to 0, 1.5, 3.0 or 6.0 mg l–1 BP‐2 until 2 months post‐metamorphosis. Overt toxicity was evident throughout the exposure period in the 6.0 mg l–1 treatment due to elevated mortality rates and observed liver and kidney pathologies. Concentration‐dependent increases in severity of thyroid follicular cell hypertrophy and hyperplasia occurred in larval tadpoles indicating BP‐2‐induced impacts on the thyroid axis. Additionally, gonads were impacted in all treatments with some genetic males showing both testis and ovary tissues (1.5 mg l–1) and 100% of the genetic males in the 3.0 and 6.0 mg l−1 treatments experiencing complete male‐to‐female sex reversal. Concentration‐dependent vitellogenin induction occurred in both genders with associated accumulations of protein in the livers, kidneys and gonads, which was likely vitellogenin and other estrogen‐responsive yolk proteins. This is the first study that demonstrates the endocrine effects of this mixed mode of action chemical in an amphibian species and demonstrates the utility of the LAGDA design for supporting chemical risk assessment. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-05-30T21:20:47.46234-05:0
      DOI: 10.1002/jat.3336
  • Triclosan is a mitochondrial uncoupler in live zebrafish
    • Authors: Juyoung Shim; Lisa M. Weatherly, Richard H. Luc, Maxwell T. Dorman, Andy Neilson, Ryan Ng, Carol H. Kim, Paul J. Millard, Julie A. Gosse
      First page: 1662
      Abstract: Triclosan (TCS) is a synthetic antimicrobial agent used in many consumer goods at millimolar concentrations. As a result of exposure, TCS has been detected widely in humans. We have recently discovered that TCS is a proton ionophore mitochondrial uncoupler in multiple types of living cells. Here, we present novel data indicating that TCS is also a mitochondrial uncoupler in a living organism: 24-hour post-fertilization (hpf) zebrafish embryos. These experiments were conducted using a Seahorse Bioscience XFe 96 Extracellular Flux Analyzer modified for bidirectional temperature control, using the XF96 spheroid plate to position and measure one zebrafish embryo per well. Using this method, after acute exposure to TCS, the basal oxygen consumption rate (OCR) increases, without a decrease in survival or heartbeat rate. TCS also decreases ATP-linked respiration and spare respiratory capacity and increases proton leak: all indicators of mitochondrial uncoupling. Our data indicate, that TCS is a mitochondrial uncoupler in vivo, which should be taken into consideration when assessing the toxicity and/or pharmaceutical uses of TCS. This is the first example of usage of a Seahorse Extracellular Flux Analyzer to measure bioenergetic flux of a single zebrafish embryo per well in a 96-well assay format. The method developed in this study provides a high-throughput tool to identify previously unknown mitochondrial uncouplers in a living organism. Copyright © 2016 John Wiley & Sons, Ltd.
      PubDate: 2016-03-28T06:00:41.219016-05:
      DOI: 10.1002/jat.3311
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