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Most Cited Mutation Research - Genetic Toxicology and Environmental Mutagenesis Articles
The most cited articles published since 2011, extracted from Scopus.
Induction of oxidative stress, DNA damage and apoptosis in mouse liver after sub-acute oral exposure to zinc oxide nanoparticlesVyom Sharma | Poonam Singh | Alok K. Pandey | Alok Dhawan
Volume 745, Issues 1-2, June 2012, Pages 84-91
Zinc oxide (ZnO) nanoparticles are finding applications in a wide range of products including cosmetics, food packaging, imaging, etc. This increases the likelihood of human exposure to these nanoparticles through dermal, inhalation and oral routes. Presently, the majority of the studies concerning ZnO nanoparticle toxicity have been conducted using . in vitro systems which lack the complex cell-cell, cell-matrix interactions and hormonal effects found in the . in vivo scenario. The present . in vivo study in mice was aimed at investigating the oral toxicity of ZnO nanoparticles. Our results showed a significant accumulation of nanoparticles in the liver leading to cellular injury after sub-acute oral exposure of ZnO nanoparticles (300. mg/kg) for 14 consecutive days. This was evident by the elevated alanine aminotransferase (ALT) and alkaline phosphatase (ALP) serum levels and pathological lesions in the liver. ZnO nanoparticles were also found to induce oxidative stress indicated by an increase in lipid peroxidation. The DNA damage in the liver and kidney cells of mice was evaluated by the Fpg-modified Comet assay which revealed a significant (. p<. 0.05) increase in the Fpg-specific DNA lesions in liver indicating oxidative stress as the cause of DNA damage. The TUNEL assay revealed an induction of apoptosis in the liver of mice exposed to ZnO nanoparticles compared to the control. Our results conclusively demonstrate that sub-acute oral exposure to ZnO nanoparticles in mice leads to an accumulation of nanoparticles in the liver causing oxidative stress mediated DNA damage and apoptosis. These results also suggest the need for a complete risk assessment of any new engineered nanoparticle before its arrival into the consumer market. © 2011 Elsevier B.V.
A core in vitro genotoxicity battery comprising the Ames test plus the in vitro micronucleus test is sufficient to detect rodent carcinogens and in vivo genotoxinsDavid Kirkland | Lesley Reeve | David Gatehouse | Philippe Vanparys
Volume 721, Issue 1, March 2011, Pages 27-73
In vitro genotoxicity testing needs to include tests in both bacterial and mammalian cells, and be able to detect gene mutations, chromosomal damage and aneuploidy. This may be achieved by a combination of the Ames test (detects gene mutations) and the in vitro micronucleus test (MNvit), since the latter detects both chromosomal aberrations and aneuploidy. In this paper we therefore present an analysis of an existing database of rodent carcinogens and a new database of in vivo genotoxins in terms of the in vitro genotoxicity tests needed to detect their in vivo activity. Published in vitro data from at least one test system (most were from the Ames test) were available for 557 carcinogens and 405 in vivo genotoxins. Because there are fewer publications on the MNvit than for other mammalian cell tests, and because the concordance between the MNvit and the in vitro chromosomal aberration (CAvit) test is so high for clastogenic activity, positive results in the CAvit test were taken as indicative of a positive result in the MNvit where there were no, or only inadequate data for the latter. Also, because Hprt and Tk loci both detect gene-mutation activity, a positive Hprt test was taken as indicative of a mouse-lymphoma Tk assay (MLA)-positive, where there were no data for the latter. Almost all of the 962 rodent carcinogens and in vivo genotoxins were detected by an in vitro battery comprising Ames. +. MNvit. An additional 11 carcinogens and six in vivo genotoxins would apparently be detected by the MLA, but many of these had not been tested in the MNvit or CAvit tests. Only four chemicals emerge as potentially being more readily detected in MLA than in Ames. +. MNvit - benzyl acetate, toluene, morphine and thiabendazole - and none of these are convincing cases to argue for the inclusion of the MLA in addition to Ames. +. MNvit. Thus, there is no convincing evidence that any genotoxic rodent carcinogens or in vivo genotoxins would remain undetected in an in vitro test battery consisting of Ames. +. MNvit. © 2011 Elsevier B.V.
Genotoxic effects of silver nanoparticles stimulated by oxidative stress in human normal bronchial epithelial (BEAS-2B) cellsHa Ryong Kim | Mi Jie Kim | Soo Yeun Lee | Seung Min Oh | Kyu Hyuck Chung
Volume 726, Issue 2, December 2011, Pages 129-135
Many classes of silver nanoparticles (Ag-NPs) have been synthesized and widely applied, but the genotoxicity of Ag-NPs and the factors leading to genotoxicity remain unknown. Therefore, the purpose of this study is to elucidate the genotoxic effects of Ag-NPs in lung and the role of oxidative stress on the genotoxic effects of Ag-NPs. For this, Ag-NPs were completely dispersed in medium by sonication and filtration. The Ag-NPs dispersed in medium were 43-260. nm in size. We observed distinct uptake of Ag-NPs into BEAS-2B cells. The Ag-NPs aggregates were wrapped with an endocytic vesicle within the cytoplasm and nucleus of BEAS-2B cells. In the comet assay and micronucleus (MN) assay for BEAS-2B cells, Ag-NPs stimulated DNA breakage and MN formation in a dose-dependent manner. The genotoxic effect of Ag-NPs was partially blocked by scavengers. In particular, of the scavengers tested, superoxide dismutase most significantly blocked the genotoxic effects in both the cytokinesis-block MN assay and the comet assay. In the modified comet assay, Ag-NPs induced a significant increase in oxidative DNA damage. Furthermore, in the oxidative stress assay, Ag-NPs significantly increased the reactive oxygen radicals. These results suggest that Ag-NPs have genotoxic effects in BEAS-2B cells and that oxidative stress stimulated by Ag-NPs may be an important factor in their genotoxic effects. © 2011 Elsevier B.V.
Effects of single and multi walled carbon nanotubes on macrophages: Cyto and genotoxicity and electron microscopyMaria Laura Di Giorgio | Sebastiano Di Bucchianico | Anna Maria Ragnelli | Pierpaolo Aimola | Sandro Santucci | Anna Poma
Volume 722, Issue 1, May 2011, Pages 20-31
Production of nanotechnology-based materials is increasing worldwide: it is essential to evaluate their potential toxicity. Among these nanomaterials, carbon nanotubes (CNTs) have tremendous potential in many areas of research and applications. We have investigated the cyto- and genotoxic effects of single and multi-walled CNTs (SWCNTs, MWCNTs) and carbon black (CB) on the mouse macrophage cell line RAW 264.7. Specifically we have investigated inflammatory response, release of tumor necrosis factor-α (TNF-α), intracellular reactive oxygen species (ROS) production, cell death (both necrosis and apoptosis), chromosomal aberrations and cellular ultrastructural alteration caused by CB, MWCNTs and SWCNTs. Our data confirm that both CNTs and CB are cyto and geno-toxic to RAW 264.7 mouse macrophages. CNTs exposure induced ROS release, necrosis and chromosomal aberrations but did not cause an inflammatory response. In addition CNTs induce ultrastructural damage and apoptosis. CNTs penetrate the cell membrane and individual MWCNTs are seen associated with the nuclear envelope. © 2011 Elsevier B.V.
Cytotoxic and genotoxic effects of multi-wall carbon nanotubes on human umbilical vein endothelial cells in vitroYuan Yuan Guo | Jun Zhang | Yi Fan Zheng | Jun Yang | Xin Qiang Zhu
Volume 721, Issue 2, April 2011, Pages 184-191
Carbon nanomaterials have multiple applications in various areas. However, it has been suggested that exposure to nanoparticles may be a risk for the development of vascular diseases due to injury and dysfunction of the vascular endothelium. Therefore, in the present study, the cytotoxic and genotoxic effects of multi-wall carbon nanotubes (MWCNTs) on human umbilical vein endothelial cells (HUVECs) were evaluated. Optical and transmission electronic microscopy (TEM) study showed that MWCNTs were able to enter cells rapidly, distribute in the cytoplasm and intracellular vesicles and induce morphological changes. Exposure to MWCNTs reduced the viability of HUVECs, and induced apoptosis in HUVECs. Furthermore, MWCNTs could cause DNA damage as indicated by the formation of γH2AX foci. MWCNTs also affected cellular redox status, e.g., increasing intracellular reactive oxygen species (ROS) and malondialdehyde (MDA) levels, as well as altering superoxide dismutase (SOD) activity and glutathione peroxidase (GSH-Px) levels. On the other hand, the free radical scavenger N-acetyl- l-cysteine (NAC) preincubation can inhibit the cytotoxic and genotoxic effects of MWCNTs. Taken together, these results demonstrated that MWCNTs could induce cytotoxic and genotoxic effects in HUVECs, probably through oxidative damage pathways. © 2011 Elsevier B.V.
Reduction of misleading (" false") positive results in mammalian cell genotoxicity assays. I. Choice of cell typePaul Fowler | Katie Smith | Jamie Young | Laura Jeffrey | David Kirkland | Stefan Pfuhler | Paul Carmichael
Volume 742, Issues 1-2, February 2012, Pages 11-25
Current in vitro mammalian cell genotoxicity assays show a high rate of positive results, many of which are misleading when compared with in vivo genotoxicity or rodent carcinogenicity data. P53-deficiency in many of the rodent cell lines may be a key factor in this poor predictivity. As part of an European Cosmetics Industry Association initiative for improvement of in vitro mammalian cell assays, we have compared several rodent cell lines (V79, CHL, CHO) with p53-competent human peripheral blood lymphocytes (HuLy), TK6 human lymphoblastoid cells, and the human liver cell line, HepG2. We have compared in vitro micronucleus (MN) induction following treatment with 19 compounds that were accepted as producing misleading or " false" positive results in in vitro mammalian cell assays [6]. Of these, six chemicals (2-ethyl-1,3-hexandiol, benzyl alcohol, urea, sodium saccharin, sulfisoxazole and isobutyraldehyde) were not toxic and did not induce any MN at concentrations up to 10. mM. d,l-Menthol and ethionamide induced cytotoxicity, but did not induce MN. o-Anthranilic acid was not toxic and did not induce MN in V79, CHL, CHO, HuLy and HepG2 cells up to 10. mM. Toxicity was induced in TK6 cells, although there were no increases in MN frequency up to and above the 55% toxicity level. The other 10 chemicals (1,3-dihydroxybenzene, curcumin, propyl gallate, p-nitrophenol, ethyl acrylate, eugenol, tert-butylhydroquinone, 2,4-dichlorophenol, sodium xylene sulfonate and phthalic anhydride) produced cytotoxicity in at least one cell type, and were evaluated further for MN induction in most or all of the cell types listed above. All these chemicals induced MN at concentrations <10. mM, with levels of cytotoxicity below 60% (measured as the replication index) in at least one cell type. The rodent cell lines (V79, CHO and CHL) were consistently more susceptible to cytotoxicity and MN induction than p53-competent cells, and are therefore more susceptible to giving misleading positive results. These data suggest that a reduction in the frequency of misleading positive results can be achieved by careful selection of the mammalian cell type for genotoxicity testing. © 2011 Elsevier B.V.
Single-walled carbon nanotube-induced mitotic disruptionL. M. Sargent | A. F. Hubbs | S. H. Young | M. L. Kashon | C. Z. Dinu | J. L. Salisbury | S. A. Benkovic | D. T. Lowry | A. R. Murray | E. R. Kisin | K. J. Siegrist | L. Battelli | J. Mastovich | J. L. Sturgeon | K. L. Bunker | A. A. Shvedova | S. H. Reynolds
Volume 745, Issues 1-2, June 2012, Pages 28-37
Carbon nanotubes were among the earliest products of nanotechnology and have many potential applications in medicine, electronics, and manufacturing. The low density, small size, and biological persistence of carbon nanotubes create challenges for exposure control and monitoring and make respiratory exposures to workers likely. We have previously shown mitotic spindle aberrations in cultured primary and immortalized human airway epithelial cells exposed to 24, 48 and 96μg/cm 2 single-walled carbon nanotubes (SWCNT). To investigate mitotic spindle aberrations at concentrations anticipated in exposed workers, primary and immortalized human airway epithelial cells were exposed to SWCNT for 24-72h at doses equivalent to 20 weeks of exposure at the Permissible Exposure Limit for particulates not otherwise regulated. We have now demonstrated fragmented centrosomes, disrupted mitotic spindles and aneuploid chromosome number at those doses. The data further demonstrated multipolar mitotic spindles comprised 95% of the disrupted mitoses. The increased multipolar mitotic spindles were associated with an increased number of cells in the G2 phase of mitosis, indicating a mitotic checkpoint response. Nanotubes were observed in association with mitotic spindle microtubules, the centrosomes and condensed chromatin in cells exposed to 0.024, 0.24, 2.4 and 24μg/cm 2 SWCNT. Three-dimensional reconstructions showed carbon nanotubes within the centrosome structure. The lower doses did not cause cytotoxicity or reduction in colony formation after 24h; however, after three days, significant cytotoxicity was observed in the SWCNT-exposed cells. Colony formation assays showed an increased proliferation seven days after exposure. Our results show significant disruption of the mitotic spindle by SWCNT at occupationally relevant doses. The increased proliferation that was observed in carbon nanotube-exposed cells indicates a greater potential to pass the genetic damage to daughter cells. Disruption of the centrosome is common in many solid tumors including lung cancer. The resulting aneuploidy is an early event in the progression of many cancers, suggesting that it may play a role in both tumorigenesis and tumor progression. These results suggest caution should be used in the handling and processing of carbon nanotubes. © 2012.
In vitro genotoxicity testing strategy for nanomaterials and the adaptation of current OECD guidelinesS. H. Doak | B. Manshian | G. J S Jenkins | N. Singh
Volume 745, Issues 1-2, June 2012, Pages 104-111
There is a pressing requirement to define a hazard identification and risk management strategy for nanomaterials due to the rapid growth in the nanotechnology industry and their promise of life-style revolutions through the development of wide-ranging nano-containing consumer products. Consequently, a battery of well defined and appropriate . in vitro assays to assess a number of genotoxicity endpoints is required to minimise extensive and costly . in vivo testing. However, the validity of the established protocols in current OECD recognised genotoxicity assays for nanomaterials is currently being questioned. In this report, we therefore consider the . in vitro OECD genotoxicity test battery including the Ames, micronucleus and HPRT forward mutation assays, and their potential role in the safety assessment of nanomaterial induced DNA damage . in vitro. © 2011 Elsevier B.V.
In vitro and in vivo genotoxicity of silver nanoparticlesManosij Ghosh | Manivannan J | Sonali Sinha | Anirban Chakraborty | Sanjaya Kumar Mallick | Maumita Bandyopadhyay | Anita Mukherjee
Volume 749, Issues 1-2, December 2012, Pages 60-69
The biocidal effect of silver nanoparticles (Ag-np) has resulted in their incorporation into consumer products. While the population exposed to Ag-np continues to increase with ever new applications, Ag-np remains a controversial research area with regard to their toxicity in biological systems. Here a genotoxic and cytotoxic approach was employed to elucidate the activity of Ag-np in vitro and in vivo. Characterization of Ag-np using scanning electron microscopy revealed a size range of 90-180. nm. Cytotoxic potential of Ag-np was evaluated in human lymphocytes via cell viability assay (Trypan blue dye exclusion method, MTT and WST assay). The uptake and incorporation of Ag-np into the lymphocytes was confirmed by flow cytometry. Additionally apoptosis (AnnexinV-FITC-PI staining) and DNA strand breaks (comet assay) in human lymphocytes revealed that Ag-np at concentration 25 μg/ml can cause genotoxicity. In vivo experiments on plants (Allium cepa and Nicotiana tabacum) and animal (Swiss albino male mice) showed impairment of nuclear DNA. Induction of oxidative stress was also studied. The DNA damage and chromosomal aberrations raise the concern about the safety associated with applications of the Ag-np. A single ip administration of Ag-np gave a significant (P≤. 0.05) increase in the frequency of aberrant cells and Tail DNA percent at concentrations 10. mg/kg body weight and above. Results of comet assay in A. cepa and N. tabacum demonstrated that the genotoxic effect of Ag-np was more pronounced in root than shoot/leaf of the plants. The present study indicated a good correlation between the in vitro and in vivo experiments. Therefore the biological applications employing Ag-np should be given special attention besides adapting the antimicrobial potential. © 2012 Elsevier B.V.
Organic extracts of urban air pollution particulate matter (PM2.5)-induced genotoxicity and oxidative stress in human lung bronchial epithelial cells (BEAS-2B cells)Seung Min Oh | Ha Ryong Kim | Yong Joo Park | Soo Yeun Lee | Kyu Hyuck Chung
Volume 723, Issue 2, August 2011, Pages 142-151
Traffic is a major source of particulate matter (PM), and ultrafine particulates and traffic intensity probably contribute significantly to PM-related health effects. As a strong relationship between air pollution and motor vehicle-originated pollutants has been shown to exist, air pollution genotoxicity studies of urban cities are steadily increasing. In Korea, the death rate caused by lung cancer is the most rapidly increased cancer death rate in the past 10 years. In this study, genotoxicity of PM2.5 (<2.5μm in aerodynamic diameter particles) collected from the traffic area in Suwon City, Korea, was studied using cultured human lung bronchial epithelial cells (BEAS-2B) as a model system for the potential inhalation health effects. Organic extract of PM2.5 (CE) generated significant DNA breakage and micronucleus formation in a dose-dependent manner (1μg/cm3-50μg/cm3). In the acid-base-neutral fractionation of PM2.5, neutral samples including the aliphatic (F3), aromatic (F4) and slightly polar (F5) fractions generated significant DNA breakage and micronucleus formation. These genotoxic effects were significantly blocked by scavenging agents [superoxide dismutase (SOD), sodium selenite (SS), mannitol (M), catalase (CAT)]. In addition, in the modified Comet assay using endonucleases (FPG and ENDOIII), CE and its fractions (F3, F4, and F5) increased DNA breakage compared with control groups, indicating that CE and fractions of PM2.5 induced oxidative DNA damage. These results clearly suggest that PM2.5 collected in the Suwon traffic area has genotoxic effects and that reactive oxygen species may play a distinct role in these effects. In addition, aliphatic/chlorinated hydrocarbons, PAH/alkylderivatives, and nitro-PAH/ketones/quinones may be important causative agents of the genotoxic effects. © 2011 Elsevier B.V.
Small molecules with big effects: The role of the microRNAome in cancer and carcinogenesisIgor Koturbash | Franz J. Zemp | Igor Pogribny | Olga Kovalchuk
Volume 722, Issue 2, June 2011, Pages 94-105
Small non-coding RNAs-microRNAs, are potent negative regulators of gene expression. MicroRNAs are involved in multiple biological processes, metabolic regulation, including cell proliferation, differentiation, and programmed cell death. Since the dysregulation of these processes is a hallmark of cancer, microRNAs can be viewed as major contributors to the pathogenesis of cancer, including initiation and progression of cancer. This review focuses on microRNA biogenesis and function, and their role in cancer, metastasis, drug resistance, and tumorigenesis. © 2010 Elsevier B.V.
Towards a more reliable comet assay: Optimising agarose concentration, unwinding time and electrophoresis conditionsAmaya Azqueta | Kristine B. Gutzkow | Gunnar Brunborg | Andrew R. Collins
Volume 724, Issues 1-2, September 2011, Pages 41-45
The comet assay is now the method of choice for measuring most kinds of DNA damage in cells. However, due to the lack of a standardised protocol inter-laboratory comparisons are of limited value. The aim of this paper is to demonstrate how small changes in comet-assay variables may significantly affect the results. We examined the effect of varying agarose concentrations, alkaline unwinding time, electrophoresis time, voltage and current, by use of two cell types, viz. human peripheral blood lymphocytes and the lymphoblastoid cell line TK-6. All these variables have marked effects on assay performance and, therefore, on the determination of DNA damage. Here we identify factors of particular importance. © 2011 Elsevier B.V.
Investigation of genotoxic and cytotoxic effects of micro- and nanosized titanium dioxide in six organs of mice in vivoLyudmila P. Sycheva | Vjacheslav S. Zhurkov | Valentina V. Iurchenko | Natalia O. Daugel-Dauge | Maria A. Kovalenko | Elena K. Krivtsova | Andrey D. Durnev
Volume 726, Issue 1, November 2011, Pages 8-14
Titanium dioxide is manufactured worldwide in large quantities for use in a wide range of applications including as food additives, in cosmetics and pigments for coloring ingested and externally applied drugs. Although TiO 2 is chemically inert it can cause negative health effects, such as lung cancer in rats. However, the mechanisms involved in TiO 2-induced genotoxicity and carcinogenicity have not been clearly defined and are poorly studied in vivo. In the present research genotoxicity and carcinogenicity of titanium dioxide were studied in a mouse model. We treated CBAB6F1 mice by oral gavage with titanium dioxide particles (microsized, TDM, 160nm; nanosized, TDN, 33nm) in doses of 40, 200 and 1000mg/kg bw, daily for seven days. Genotoxic effects were analyzed in the cells of brain, liver and bone marrow by means of the Comet assay and in the cells of bone marrow, forestomach, colon and testis with a poly-organ karyological assay (analysis of micronuclei, nuclear protrusions, atypical nuclei, multinucleated cells, mitotic and/or apoptotic index). TDM induced DNA-damage and micronuclei in bone-marrow cells and TDN induced DNA-damage in the cells of bone marrow and liver. TDM and TDN increased the mitotic index in forestomach and colon epithelia, the frequency of spermatids with two and more nuclei, and apoptosis in forestomach (only TDN) and testis. This is one of the first poly-organ studies of TDM- and TDN-induced genotoxicity in vivo in mice. These effects are caused by a secondary genotoxic mechanism associated with inflammation and/or oxidative stress. Given the increasing use of TiO 2 nanoparticles, these findings indicate a potential health hazard associated with exposure to TiO 2 particles. © 2011 Elsevier B.V.
When pigs fly: Immunomagnetic separation facilitates rapid determination of Pig-a mutant frequency by flow cytometric analysisStephen D. Dertinger | Steven M. Bryce | Souk Phonethepswath | Svetlana L. Avlasevich
Volume 721, Issue 2, April 2011, Pages 163-170
In vivo mutation assays based on the Pig-a null phenotype, that is, the absence of cell surface glycosylphosphatidylinositol (GPI) anchored proteins such as CD59, have been described. This work has been accomplished with hematopoietic cells, most often rat peripheral blood erythrocytes (RBCs) and reticulocytes (RETs). The current report describes new sample processing procedures that dramatically increase the rate at which cells can be evaluated for GPI anchor deficiency. This new method was applied to blood specimens from vehicle, 1,3-propane sultone, melphalan, and N-ethyl- N-nitrosourea treated Sprague Dawley rats. Leukocyte- and platelet-depleted blood samples were incubated with anti-CD59-phycoerythrin (PE) and anti-CD61-PE, and then mixed with anti-PE paramagnetic particles and Counting Beads (i.e., fluorescent microspheres). An aliquot of each specimen was stained with SYTO 13 and flow cytometric analysis was performed to determine RET percentage, RET:Counting Bead ratio, and RBC:Counting Bead ratio. The major portion of these specimens were passed through ferromagnetic columns that were suspended in a magnetic field, thereby depleting each specimen of wild-type RBCs (and platelets) based on their association with anti-PE paramagnetic particles. The eluates were concentrated via centrifugation and the resulting suspensions were stained with SYTO 13 and analyzed on the flow cytometer to determine mutant phenotype RET:Counting Bead and mutant phenotype RBC:Counting Bead ratios. The ratios obtained from pre- and post-column analyses were used to derive mutant phenotype RET and mutant phenotype RBC frequencies. Results from vehicle control and genotoxicant-treated rats are presented that indicate the scoring system is capable of returning reliable mutant phenotype cell frequencies. Using this wild-type cell depletion strategy, it was possible to interrogate ≥3 million RETs and ≥100 million RBCs per rat in approximately 7. min. Beyond considerably enhancing the throughput capacity of the analytical platform, these blood-processing procedures were also shown to enhance the precision of the measurements. © 2011 Elsevier B.V.
Improvement of in vivo genotoxicity assessment: Combination of acute tests and integration into standard toxicity testingAndreas Rothfuss | Masamitu Honma | Andreas Czich | Marilyn J. Aardema | Brian Burlinson | Sheila Galloway | Shuichi Hamada | David Kirkland | Robert H. Heflich | Jonathan Howe | Madoka Nakajima | Mike O'Donovan | Ulla Plappert-Helbig | Catherine Priestley | Leslie Recio | Maik Schuler | Yoshifumi Uno | Hans Jörg Martus
Volume 723, Issue 2, August 2011, Pages 108-120
A working group convened at the 2009 5th IWGT to discuss possibilities for improving in vivo genotoxicity assessment by investigating possible links to standard toxicity testing. The working group considered: (1) combination of acute micronucleus (MN) and Comet assays into a single study, (2) integration of MN assays into repeated-dose toxicity (RDT) studies, (3) integration of Comet assays into RDT studies, and (4) requirements for the top dose when integrating genotoxicity measurements into RDT studies. The working group reviewed current requirements for in vivo genotoxicity testing of different chemical product classes and identified opportunities for combination and integration of genotoxicity endpoints for each class. The combination of the acute in vivo MN and Comet assays was considered by the working group to represent a technically feasible and scientifically acceptable alternative to conducting independent assays. Two combination protocols, consisting of either a 3- or a 4-treament protocol, were considered equally acceptable. As the integration of MN assays into RDT studies had already been discussed in detail in previous IWGT meetings, the working group focussed on factors that could affect the results of the integrated MN assay, such as the possible effects of repeated bleeding and the need for early harvests. The working group reached the consensus that repeated bleeding at reasonable volumes is not a critical confounding factor for the MN assay in rats older than 9 weeks of age and that rats bled for toxicokinetic investigations or for other routine toxicological purposes can be used for MN analysis. The working group considered the available data as insufficient to conclude that there is a need for an early sampling point for MN analysis in RDT studies, in addition to the routine determination at terminal sacrifice. Specific scenarios were identified where an additional early sampling can have advantages, e.g., for compounds that exert toxic effects on hematopoiesis, including some aneugens. For the integration of Comet assays into RDT studies, the working group reached the consensus that, based upon the limited amount of data available, integration is scientifically acceptable and that the liver Comet assay can complement the MN assay in blood or bone marrow in detecting in vivo genotoxins. Practical issues need to be considered when conducting an integrated Comet assay study. Freezing of tissue samples for later Comet assay analysis could alleviate logistical problems. However, the working group concluded that freezing of tissue samples can presently not be recommended for routine use, although it was noted that results from some laboratories look promising. Another discussion topic centred around the question as to whether tissue toxicity, which is more likely observed in RDT than in acute toxicity studies, would affect the results of the Comet assay. Based on the available data from in vivo studies, the working group concluded that there are no clear examples where cytotoxicity, by itself, generates increases or decreases in DNA migration. The working group identified the need for a refined guidance on the use and interpretation of cytotoxicity methods used in the Comet assay, as the different methods used generally lead to inconsistent conclusions. Since top doses in RDT studies often are limited by toxicity that occurs only after several doses, the working group discussed whether the sensitivity of integrated genotoxicity studies is reduced under these circumstances. For compounds for which in vitro genotoxicity studies yielded negative results, the working group reached the consensus that integration of in vivo genotoxicity endpoints (typically the MN assay) into RDT studies is generally acceptable. If in vitro genotoxicity results are unavailable or positive, consensus was reached that the maximum tolerated dose (MTD) is acceptable as the top dose in RDT studies in many cases, such as when the RDT study MTD or exposure is close (50% or greater) to an acute study MTD or exposure. Finally, the group agreed that exceptions to this general rule might be acceptable, for example when human exposure is lower than the preclinical exposure by a large margin. © 2010 Elsevier B.V.
Amorphous silica nanoparticles do not induce cytotoxicity, cell transformation or genotoxicity in Balb/3T3 mouse fibroblastsChiara Uboldi | Guido Giudetti | Francesca Broggi | Douglas Gilliland | Jessica Ponti | François Rossi
Volume 745, Issues 1-2, June 2012, Pages 11-20
Although amorphous silica nanoparticles (aSiO 2NPs) are believed to be non-toxic and are currently used in several industrial and biomedical applications including cosmetics, food additives and drug delivery systems, there is still no conclusive information on their cytotoxic, genotoxic and carcinogenic potential. For this reason, this work has investigated the effects of aSiO 2NPs on Balb/3T3 mouse fibroblasts, focusing on cytotoxicity, cell transformation and genotoxicity. Results obtained using aSiO 2NPs, with diameters between 15nm and 300nm and exposure times up to 72h, have not shown any cytotoxic effect on Balb/3T3 cells as measured by the MTT test and the Colony Forming Efficiency (CFE) assay. Furthermore, aSiO 2NPs have induced no morphological transformation in Balb/3T3 cells and have not resulted in genotoxicity, as shown by Cell Transformation Assay (CTA) and Micronucleus (MN) assay, respectively. To understand whether the absence of any toxic effect could result from a lack of internalization of the aSiO 2NPs by Balb/3T3 cells, we have investigated the uptake and the intracellular distribution following exposure to 85nm fluorescently-labelled aSiO 2NPs. Using fluorescence microscopy, it was observed that fluorescent aSiO 2NPs are internalized and located exclusively in the cytoplasmic region. In conclusion, we have demonstrated that although aSiO 2NPs are internalized in vitro by Balb/3T3 mouse fibroblasts, they do not trigger any cytotoxic or genotoxic effect and do not induce morphological transformation, suggesting that they might be a useful component in industrial applications. © 2011 Elsevier B.V.
Exposure of pregnant mice to carbon black by intratracheal instillation: Toxicogenomic effects in dams and offspringPetra Jackson | Karin S. Hougaard | Ulla Vogel | Dongmei Wu | Lorraine Casavant | Andrew Williams | Mike Wade | Carole L. Yauk | Håkan Wallin | Sabina Halappanavar
Volume 745, Issues 1-2, June 2012, Pages 73-83
Exposure to nanomaterials (NM) during sensitive developmental stages may predispose organisms to diseases later in life. However, direct translocation of NM from mother to fetus through the placenta is limited. The present study tests the hypothesis that pulmonary exposure to NM and NM-induced response, such as inflammation during gestation, leads to secondary effects in the fetus. Time-mated C57BL/6BomTac mice were exposed by intratracheal instillation to vehicle (Nanopure water) or one of three concentrations (2.75, 13.5 or 67. μg in 40. μl Nanopure water) of carbon black Printex 90 (CB) on gestational days 7, 10, 15 and 18, to final cumulative doses of 11, 54 or 268. μg/animal. Samples from a subset of male and female newborns were collected on postnatal day 2 (4. days after the last maternal exposure) and from dams 26 to 27. days post-exposure (post-weaning period). Histopathology, DNA microarrays, pathway-specific RT-PCR arrays, focussed RT-PCR, and tissue protein analysis were employed to characterize pulmonary response in dams exposed to CB during pregnancy. Hepatic gene expression in newborns was interpreted in light of the observed biological responses and gene expression changes arising in the lungs of dams following CB exposure. Although retention of CB particles was observed in dams from both the medium and the high dose groups, neutrophil-marked inflammation and altered expression of several cytokines and chemokines, both at the transcriptional and tissue protein levels, was significant only in the high dose group. Analysis of newborn livers by DNA microarrays revealed that female offspring were more sensitive to maternal exposure than male offspring. Cellular signalling, inflammation, cell cycle and lipid metabolism were among the biological pathways affected in female offspring. Males, however, responded with subtle changes in metabolism-related genes. Further investigation is required to determine the long-term health consequences of the gene expression changes in offspring and response to environmental stresses. © 2011.
In vitro genotoxicity test approaches with better predictivity: Summary of an IWGT workshopStefan Pfuhler | Mick Fellows | Jan Van Benthem | Raffaella Corvi | Rodger Curren | Kerry Dearfield | Paul Fowler | Roland Frötschl | Azeddine Elhajouji | Ludovic Le Hégarat | Toshio Kasamatsu | Hajime Kojima | Gladys Ouédraogo | Andrew Scott | Günter Speit
Volume 723, Issue 2, August 2011, Pages 101-107
Improving current in vitro genotoxicity tests is an ongoing task for genetic toxicologists. Further, the question on how to deal with positive in vitro results that are demonstrated to not predict genotoxicity or carcinogenicity potential in rodents or humans is a challenge. These two aspects were addressed at the 5th International Workshop on Genotoxicity Testing (IWGT) held in Basel, Switzerland, on August 17-19, 2009. The objectives of the working group (WG) were to make recommendations on the use of cell types or lines, if possible, and to provide evaluations of promising new approaches. Results obtained in rodent cell lines with impaired p53 function (L5178Y, V79, CHL and CHO cells) and human p53-competent cells (peripheral blood lymphocytes, TK6 and HepG2 cells) suggest that a reduction in the percentage of non-relevant positive results for carcinogenicity prediction can be achieved by careful selection of cells used without decreasing the sensitivity of the assays. Therefore, the WG suggested using p53- competent - preferably human - cells in in vitro micronucleus or chromosomal aberration tests. The use of the hepatoma cell line HepaRG for genotoxicity testing was considered promising since these cells possess better phase I and II metabolizing potential compared to cell lines commonly used in this area and may overcome the need for the addition of S9. For dermally applied compounds, the WG agreed that in vitro reconstructed skin models, once validated, will be useful to follow up on positive results from standard in vitro assays as they resemble the properties of human skin (barrier function, metabolism). While the reconstructed skin micronucleus assay has been shown to be further advanced, there was also consensus that the Comet assay should be further evaluated due to its independence from cell proliferation and coverage of a wider spectrum of DNA damage. © 2011 Elsevier B.V.
Antiproliferative effects of goniothalamin on Ca9-22 oral cancer cells through apoptosis, DNA damage and ROS inductionChing Yu Yen | Chien Chih Chiu | Rou Wen Haung | Chi Chen Yeh | Kuang Jing Huang | Kuo Feng Chang | You Cheng Hseu | Fang Rong Chang | Hsueh Wei Chang | Yang Chang Wu
Volume 747, Issue 2, September 2012, Pages 253-258
Goniothalamin (GTN), a plant bioactive styryl-lactone, is a natural product with potent anti-tumorigenesis effects for several types of cancer. Nonetheless, the anticancer effect of GTN has not been examined in oral cancer. The present study was designed to evaluate its potential anticancer effects in an oral squamous cell carcinoma (OSCC) model and to determine the possible mechanisms with respect to apoptosis, DNA damage, reactive oxygen species (ROS) induction, and mitochondrial membrane potential. Our data demonstrated that cell proliferation was significantly inhibited by GTN in Ca9-22 OSCC cancer cells in concentration- and time-dependent manners (p<0.05). For cell cycle and apoptotic effects of GTN-treated Ca9-22 cancer cells, the sub-G1 population and annexin V-intensity significantly increased in a concentration-dependent manner (p<0.001). For the analysis of DNA double strand breaks, γH2AX intensity significantly increased in GTN-treated Ca9-22 cancer cells in concentration-response relationship (p<0.05). Moreover, GTN significantly induced intracellular ROS levels in Ca9-22 cancer cells in a concentration- and time-dependent manner (p<0.05). For membrane depolarization of mitochondria, the DiOC 2(3) (3,3'-diethyloxacarbocyanine iodide) intensity of GTN-treated Ca9-22 cancer cells was significantly decreased in concentration- and time-dependent relationships (p<0.001). Taken together, these results suggest that the anticancer effect of GTN against oral cancer cells is valid and GTN-induced growth inhibition and apoptosis influence the downstream cascade including ROS induction, DNA damage, and mitochondria membrane depolarization. Therefore, GTN has potential as a chemotherapeutic agent against oral cancer. © 2012 Elsevier B.V.
Genotoxicity of silver nanoparticles evaluated using the Ames test and in vitro micronucleus assayYan Li | David H. Chen | Jian Yan | Ying Chen | Roberta A. Mittelstaedt | Yongbin Zhang | Alexandru S. Biris | Robert H. Heflich | Tao Chen
Volume 745, Issues 1-2, June 2012, Pages 4-10
Silver nanoparticles (AgNPs) have antimicrobial properties, which have contributed to their widespread use in consumer products. A current issue regarding nanomaterials is the extent to which existing genotoxicity assays are useful for evaluating the risks associated with their use. In this study, the genotoxicity of 5. nm AgNPs was assessed using two standard genotoxicity assays, the . Salmonella reverse mutation assay (Ames test) and the . in vitro micronucleus assay. Using the preincubation version of the Ames assay, . Salmonella strains TA102, TA100, TA1537, TA98, and TA1535 were treated with 0.15-76.8. μg/plate of the AgNPs. Toxicity limited the doses that could be assayed to 2.4-38.4. μg/plate; no increases in mutant frequency over the vehicle control were found for the concentrations that could be assayed. Human lymphoblastoid TK6 cells were treated with 10-30. μg/ml AgNPs, and additional cells were treated with water and 0.73. gy X-rays as vehicle and positive controls. Micronucleus frequency was increased by the AgNP treatment in a dose-dependent manner. At a concentration of 30. μg/ml (with 45.4% relative population doubling), AgNPs induced a significant, 3.17-fold increase with a net increase of 1.60% in micronucleus frequency over the vehicle control, a weak positive response by our criteria. These results demonstrate that the 5. nm AgNP are genotoxic in TK6 cells. Also, the data suggest that the . in vitro micronucleus assay may be more appropriate than the Ames test for evaluating the genotoxicity of the AgNPs. © 2011.
Merging nano-genotoxicology with eco-genotoxicology: An integrated approach to determine interactive genotoxic and sub-lethal toxic effects of CSherain N. Al-Subiai | Volker M. Arlt | Patricia E. Frickers | James W. Readman | Björn Stolpe | Jamie R. Lead | A. John Moody | Awadhesh N. Jha
60 fullerenes and fluoranthene in marine mussels, Mytilus sp.
Volume 745, Issues 1-2, June 2012, Pages 92-103
Whilst there is growing concern over the potential detrimental impact of engineered nanoparticles (ENPs) on the natural environment, little is known about their interactions with other contaminants. In the present study, marine mussels (Mytilus sp.) were exposed for 3 days to C 60 fullerenes (C 60; 0.10-1mgl -1) and a model polycyclic aromatic hydrocarbon (PAH), fluoranthene (32-100μgl -1), either alone or in combination. The first two experiments were conducted by exposing the organisms to different concentrations of C 60 and fluoranthene alone, in order to determine the effects on total glutathione levels (as a measure of generic oxidative stress), genotoxicity (DNA strand breaks using Comet assay in haemocytes), DNA adduct analyses (using 32P-postlabelling method) in different organs, histopathological changes in different tissues (i.e. adductor muscle, digestive gland and gills) and physiological effects (feeding or clearance rate). Subsequently, in the third experiment, a combined exposure of C 60 plus fluoranthene (0.10mgl -1 and 32μgl -1, respectively) was carried out to evaluate all endpoints mentioned above. Both fluoranthene and C 60 on their own caused concentration-dependent increases in DNA strand breaks as determined by the Comet assay. Formation of DNA adducts however could not be detected for any exposure conditions. Combined exposure to C 60 and fluoranthene additively enhanced the levels of DNA strand breaks along with a 2-fold increase in the total glutathione content. In addition, significant accumulation of C 60 was observed in all organs, with highest levels in digestive gland (24.90±4.91μgC 60g -1ww). Interestingly, clear signs of abnormalities in adductor muscle, digestive gland and gills were observed by histopathology. Clearance rates indicated significant differences compared to the control with exposure to C 60, and C 60/fluoranthene combined treatments, but not after fluoranthene exposure alone. This study demonstrated that at the selected concentrations, both C 60 and fluoranthene evoke toxic responses and genetic damage. The combined exposure produced enhanced damage with additive rather than synergistic effects. © 2011 Elsevier B.V.
Clastogenic and mutagenic effects of bisphenol A: An endocrine disruptorDinesh Tiwari | Jyoti Kamble | Swapnil Chilgunde | Prakash Patil | Girish Maru | Dhananjay Kawle | Uma Bhartiya | Lebana Joseph | Geeta Vanage
Volume 743, Issues 1-2, March 2012, Pages 83-90
Bisphenol A (BPA) is a well-known endocrine disruptor (ED) which represents a major toxicological and public health concern due to its widespread exposure to humans. BPA has been reported to induce DNA adduct and aneuploidy in rodents. Recent studies in humans depicted its association with recurrent miscarriages and male infertility due to sperm DNA damage indicating that BPA might have genotoxic activity. Hence, the present study was designed to determine genotoxic and mutagenic effects of BPA using in-vivo and in-vitro assays. The adult male and female rats were orally administered with various doses of BPA (2.4μg, 10μg, 5. mg and 50. mg/kg. bw) once a day for six consecutive days. Animals were sacrificed, bone marrow and blood samples were collected and subjected to series of genotoxicity assay such as micronucleus, chromosome aberration and single cell gel electrophoresis (SCGE) assay respectively. Mutagenicity was determined using tester strains of Salmonella typhimurium (TA 98, TA 100 and TA 102) in the presence and absence of metabolically active microsomal fractions (S9). Further, we estimated the levels of 8-hydroxydeoxyguanosine, lipid per-oxidation and glutathione activity to decipher the potential genotoxic mechanism of BPA. We observed that BPA exposure caused a significant increase in the frequency of micronucleus (MN) in polychromatic erythrocytes (PCEs), structural chromosome aberrations in bone marrow cells and DNA damage in blood lymphocytes. These effects were observed at various doses tested except 2.4μg compared to vehicle control. We did not observe the mutagenic response in any of the tester strains tested at different concentrations of BPA. We found an increase in the level of 8-hydroxydeoxyguanosine in the plasma and increase in lipid per-oxidation and decrease in glutathione activity in liver of rats respectively which were exposed to BPA. In conclusion, the data obtained clearly documents that BPA is not mutagenic but exhibit genotoxic activity and oxidative stress could be one of the mechanisms leading to genetic toxicity. © 2012 Elsevier B.V.
Genotoxicity of inhaled nanosized TiOHanna K. Lindberg | Ghita C M Falck | Julia Catalán | Antti J. Koivisto | Satu Suhonen | Hilkka Järventaus | Elina M. Rossi | Heli Nykäsenoja | Yrjö Peltonen | Carlos Moreno | Harri Alenius | Timo Tuomi | Kai M. Savolainen | Hannu Norppa
2 in mice
Volume 745, Issues 1-2, June 2012, Pages 58-64
In vitro studies have suggested that nanosized titanium dioxide (TiO 2) is genotoxic. The significance of these findings with respect to in vivo effects is unclear, as few in vivo studies on TiO 2 genotoxicity exist. Recently, nanosized TiO 2 administered in drinking water was reported to increase, e.g., micronuclei (MN) in peripheral blood polychromatic erythrocytes (PCEs) and DNA damage in leukocytes. Induction of micronuclei in mouse PCEs was earlier also described for pigment-grade TiO 2 administered intraperitoneally. The apparent systemic genotoxic effects have been suggested to reflect secondary genotoxicity of TiO 2 due to inflammation. However, a recent study suggested that induction of DNA damage in mouse bronchoalveolar lavage (BAL) cells after intratracheal instillation of nanosized or fine TiO 2 is independent of inflammation. We examined here, if inhalation of freshly generated nanosized TiO 2 (74% anatase, 26% brookite; 5 days, 4h/day) at 0.8, 7.2, and (the highest concentration allowing stable aerosol production) 28.5mg/m 3 could induce genotoxic effects in C57BL/6J mice locally in the lungs or systematically in peripheral PCEs. DNA damage was assessed by the comet assay in lung epithelial alveolar type II and Clara cells sampled immediately following the exposure. MN were analyzed by acridine orange staining in blood PCEs collected 48h after the last exposure. A dose-dependent deposition of Ti in lung tissue was seen. Although the highest exposure level produced a clear increase in neutrophils in BAL fluid, indicating an inflammatory effect, no significant effect on the level of DNA damage in lung epithelial cells or micronuclei in PCEs was observed, suggesting no genotoxic effects by the 5-day inhalation exposure to nanosized TiO 2 anatase. Our inhalation exposure resulted in much lower systemic TiO 2 doses than the previous oral and intraperitoneal treatments, and lung epithelial cells probably received considerably less TiO 2 than BAL cells in the earlier intratracheal study. © 2011 Elsevier B.V.
Silica nanoparticles administered at the maximum tolerated dose induce genotoxic effects through an inflammatory reaction while gold nanoparticles do notThomas R. Downs | Meredith E. Crosby | Ting Hu | Shyam Kumar | Ashley Sullivan | Katherine Sarlo | Bob Reeder | Matt Lynch | Matthew Wagner | Tim Mills | Stefan Pfuhler
Volume 745, Issues 1-2, June 2012, Pages 38-50
While the collection of genotoxicity data and insights into potential mechanisms of action for nano-sized particulate materials (NPs) are steadily increasing, there is great uncertainty whether current standard assays are suitable to appropriately characterize potential risks. We investigated the effects of NPs in an . in vivo Comet/micronucleus (MN) combination assay and in an . in vitro MN assay performed with human blood. We also incorporated additional endpoints into the . in vivo study in an effort to delineate primary from secondary mechanisms. Amorphous silica NPs (15 and 55. nm) were chosen for their known reactivity, while gold nano/microparticles (2, 20, and 200. nm) were selected for their wide size range and lower reactivity. DNA damage in liver, lung and blood cells and micronuclei in circulating reticulocytes were measured after 3 consecutive intravenous injections to male Wistar rats at 48, 24 and 4. h before sacrifice. Gold nano/microparticles were negative for MN induction . in vitro and . in vivo, and for the induction of DNA damage in all tissues. Silica particles, however, caused a small but reproducible increase in DNA damage and micronucleated reticulocytes when tested at their maximum tolerated dose (MTD). No genotoxic effects were observed at lower doses, and the . in vitro MN assay was also negative. We hypothesize that silica NPs initiate secondary genotoxic effects through release of inflammatory cell-derived oxidants, similar to that described for crystalline silica (quartz). Such a mechanism is supported by the occurrence of increased neutrophilic infiltration, necrosis, and apoptotic cells in the liver, and induction of inflammatory markers TNF-α and IL-6 in plasma at the MTDs. These results were fairly consistent between silica NPs and the quartz control, thereby strengthening the argument that silica NPs may act in a similar, thresholded manner. The observed profile is supportive of a secondary genotoxicity mechanism that is driven by inflammation. © 2012 Elsevier B.V.
Early genotoxic effects in gill cells and haemocytes of Dreissena polymorpha exposed to cadmium, B[a]P and a combination of B[a]P and CdFrançoise Vincent-Hubert | Adeline Arini | Catherine Gourlay-Francé
Volume 723, Issue 1, July 2011, Pages 26-35
The aim of this study was to assess the genotoxic potential of environmentally relevant concentrations of Cd on the zebra mussel, an important freshwater sentinel organism, and to determine the stability of DNA damage in gill cells and haemocytes. The oxidative DNA damage and the co-genotoxicity of Cd in combination with B[a]P were investigated. We measured DNA damage in haemocytes and gill cells of zebra mussels exposed for 11 days to a constant concentration of Cd (10μg/L), B[a]P (10μg/L) or the two combined chemicals (10μg/L+1μg/L). Enzymatic dissociation of gills with dispase gave the lower percentage DNA in tail, compared with collagenase/dispase or collagenase. Bioaccumulation of cadmium in the soft tissues of mussels exposed to CdCl2 or CdCl2+B[a]P increased in a time-dependent manner indicating that both exposures were effective. Cd (10μg/L) is genotoxic only during the first 3 days of exposure in gill cells, while in haemocytes the genotoxicity of Cd was observed later. B[a]P (10μg/L) induced an early increase of DNA damage in gill cells (after 10h and 1 day), while in both gill cells and haemocytes, B[a]P caused a marked increase of DNA damage after 3 days of exposure. The Cd+B[a]P mixture decreased the DNA-damaging effect of Cd and B[a]P in both cell types. Cd induced an increase of DNA damage in Fpg-treated slides, indicating that Cd contributed to oxidative DNA damage. Cadmium induced a cytogenetic effect in gill cells, assessed by the number of micronuclei, throughout the duration of the exposure, while B[a]P did not induce any cytogenetic effect. B[a]P, Cd and Cd+B[a]P induced a transient increase in the number of bi-nucleated cells. Our data clearly show that gills are more sensitive to Cd and B[a]P, which makes them more suitable for future bio-monitoring studies. © 2011 Elsevier B.V.


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Sex on the Beach
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Captain Knauer (Character)
from The Longest Yard (1974)
|Photos (See all 3 | slideshow)
FilmographyEdit Credits
- The Longest Yard (2005) Played by William Fichtner
- The Longest Yard (1974) Played by Ed Lauter
... aka "The Mean Machine" - Ireland (English title) (imdb display title), UK
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Captain Knauer: Do you have any idea who's beating you out there? This was supposed to be a blow out and they're showing you up in front of the whole nation. The whole nation!
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Calm before the storm
[Verse 1]
From whence I came, express my pain
Shape and mold my frame, I go by the name of O.C
My PKA when onstage plus in real life this here's my government title
Idle to many, not the truth to naysayers
Been an avenue walker since British walkers
Way before Obama was thought to be in office
Loss of friends to jail and some dead in a coffin
Way of life and music has since gone corporate
Picture the days past now it's a whole different portrait
Me dumbin' down would be a sign of weakness
Throwin' in the towel as Victor Ortiz did
Protect yourselves at all times my guard's up
Keepin' the odds even with the flow and the bars up
The Star Child leave 'em star-struck
Knowin' an engine won't run on its own without spark plugs
Sample: Barry Forgie – Dawn Mist
Whole CD produced by Apollo Brown.
Vocals by OC. .


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HOK Chicago Office: LEED-CI Platinum
Chicago, Illinois
Having outgrown its office space and facing lease expiration, HOK Chicago enlisted the help of its own designers to assist with its relocation to a new office space in Chicago's CNA building.
Design of the 27,000-square-foot space, which is expected to become one of the only LEED CI Platinum Certified environments in Chicago, was inspired by the concepts of "collaborate," "connect," and "contrast." The new office environment provides workspace for 140 employees within a 98% open floor plan, which allows for the mobility of teams and provides maximum efficiency and effectiveness. Extensive conferencing space, a large café and informal teaming/project display areas throughout the space further facilitate employee interaction.
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Many sustainable design features were incorporated into the design, including use of salvaged materials; natural daylighting; cradle to cradle certification for workstations; and water use reduction through retrofitting or replacement of existing fixtures.
Interiors: HOK
Photography: Steve Hall - Hedrich Blessing
All rights reserved. Any unauthorized reproduction or copying is prohibited.


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The Librarian: The Curse of the Judas Chalice
- Network: TNT
- Series Premiere Date: Dec 7, 2008
- Summary: Flynn the librarian is once again trying to find an artifact, this time the Judas chalice, which can bring the dead back to life.
- Genre(s): Drama, Movie/Mini-Series, Action & Adventure
- Show Type: TV Movie
- Season 1 premiere date: Dec 7, 2008
- Episode Length: 90
- More Details and Credits »
Score distribution:
Positive: 6 out of 6
Mixed: 0 out of 6
Negative: 0 out of 6
The plot is a trifle chaotic, but the action culminates in an impressive sequence of special-effects derring-do and whooshing bloodsuckers.
The latest (and last) in the series featuring superhero librarian Flynn Carsen (Noah Wyle) packs more humor, suspense and adventure into two hours than either of its two predecessors.
It's a fun ride and Wyle has gotten a little better each time. That's why it's a shame there apparently won't be another.
Wyle is funny and charming and dispenses just enough fascinating arcane knowledge to keep things semi-educational, the special effects are great, Newhart and Curtin are always great to see, the ubiquitous Davison ("Knight Rider") is obviously having a blast as Lazlo, and really, who doesn't like a good vampire story?
In essence, Librarian is a summer movie blockbuster on a winter TV budget, and has all the virtues and flaws you'd expect from that odd pairing--including never taking itself too seriously.
The action itself is pedestrian, but as with the previous Librarian adventures, there's just enough wit around the edges to keep you watching.
Score distribution:
Positive: 1 out of 1
Mixed: 0 out of 1
Negative: 0 out of 1
Mar 6, 2016It's kind of eerie how similar this series is is to Indiana Jones movies, both in design and reception. 1st movie being original, 2nd movieIt's kind of eerie how similar this series is is to Indiana Jones movies, both in design and reception. 1st movie being original, 2nd movie being a formulaic sequel, 3rd movie going all out and expanding the lore.
This movie, like its predecessors,is full of adventure an fun, but this time adds in some depth and good character development.
My favorite so far of the Librarian movie. It's more of the same, but done very well.… Expand
- By user score


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realmagick.com The shrine of knowledge.
Didymous Judas Thomas
A selection of articles related to didymous judas thomas.
Original articles from our library related to the Didymous Judas Thomas. See Table of Contents for further available material (downloadable resources) on Didymous Judas Thomas.
- The Thinking Rede
- It would be much easier if the Rede acted like a giant Magic Eight Ball. Ask it a question, shake the ball, and wait for "yes," "no," or "absolutely!" to float to the top of the murkey window. Unfortunately, the deceivingly...
Paganism & Wicca >> The Wiccan Rede
- The Rise Of The Rede
- A Change In Emphasis The Wiccan Rede currently enjoys an extremely visible role in Paganism. As of this writing, a browser search using the term "Wiccan Rede" will generate no less than 4,000 listings. Go to a specific Web site with a "basics...
Paganism & Wicca >> The Wiccan Rede
- King James Bible: John, Chapter 20
- Chapter 20 20:1 The first day of the week cometh Mary Magdalene early, when it was yet dark, unto the sepulchre, and seeth the stone taken away from the sepulchre. 20:2 Then she runneth, and cometh to Simon Peter, and to the other disciple, whom Jesus loved,...
New Testament >> John
- Story of the Celts: The Celts of Ireland
- The Celts of Ireland [ 33 ] The Celts arrived in Ireland by 350 B.C (some say earlier) and they thrive there to this day. A claim might be made that the Celtic Irish are among the world's oldest nationality groups. Despite periods when foreigners tried in...
History & Anthropology >> Celtic & Irish
- King James Bible: Acts, Chapter 1
- Chapter 1 1:1 The former treatise have I made, O Theophilus, of all that Jesus began both to do and teach, 1:2 Until the day in which he was taken up, after that he through the Holy Ghost had given commandments unto the apostles whom he had chosen: 1:3 To...
New Testament >> The Acts
- King James Bible: Luke, Chapter 6
- Chapter 6 6:1 And it came to pass on the second sabbath after the first, that he went through the corn fields; and his disciples plucked the ears of corn, and did eat, rubbing them in their hands. 6:2 And certain of the Pharisees said unto them, Why do ye...
New Testament >> Luke
- King James Bible: Luke, Chapter 22
- Chapter 22 22:1 Now the feast of unleavened bread drew nigh, which is called the Passover. 22:2 And the chief priests and scribes sought how they might kill him; for they feared the people. 22:3 Then entered Satan into Judas surnamed Iscariot, being of the...
New Testament >> Luke
Didymous Judas Thomas is described in multiple online sources, as addition to our editors' articles, see section below for printable documents, Didymous Judas Thomas books and related discussion.
Suggested Pdf Resources
- No.44 - Middlesex Heraldry Society
- Christmas dinner- a custom we may well need to revive. Thomas Didymous is mentioned in all the Gospels, but is best remembered as the Doubting Disciple.
- www.middlesex-heraldry.org.uk
- January - Welbury
- Jan 23, 2016 Disciple Quiz Answers. From P.4.
- www.welbury.com
Suggested News Resources
- 'Allah Will Not Forgive': Shiite Leaders Threaten Saudis After Cleric Execution
- The most prominent leaders in the Shiite world have issued threatening statements against the Saudi monarchy, pledging that the House of Saud will pay a price for executing Sheikh Nimr al-Nimr, who was an influential cleric among the Saudi Shiite minority.
- 72 CE: Thomas the Apostle Is Murdered in India
- In both the Book of John, one of the Gospels of the New Testament, and in the apocryphal Acts of Thomas, Thomas is described as "Thomas, who is called Didymus," a redundancy, since "Thomas" comes from the Aramaic word teoma, meaning "twin" (in ...
- NASA Faked Mars Landings: Mars Rover Photos Were Taken In Simulated Mars
- A conspiracy theory fast gaining traction online makes the astounding claim that NASA's Curiosity and Opportunity rovers never traveled to Mars and that the images of the Martian environment being uploaded to NASA websites were actually taken on a ...
- NASA Warns Earth's Magnetic Field Weakening, Poles Shift Imminent: Reversal
- NASA has confirmed fears that recent weakening of the Earth's magnetic field indicates that we are soon going to see a reversal of Earth's magnetic poles that would cause compasses to point to the south instead of the north.
- Digital Humanities at #AARSBL15
- While BYU has few papyri of its own, one important exception is a commentary on the Psalms by Didymus the Blind, a palimpsest.
Suggested Web Resources
- The Gospel of Thomas - The Fifth (5th) Gospel - Didymos Judas
- The Gospel of Thomas - The Fifth (5th) Gospel - Didymos Judas Thomas Twin of Jesus - The Gospel According to Thomas.
- reluctant-messenger.com
- The Gnostic Apostle Thomas - "Twin" of Jesus - The Gnosis Archive
- In the Syriac-speaking culture of upper Mesopotamia and Syria the apostle was called Judas Thomas. Thomas (Tau'ma) means twin in Syriac, a form of the ...
- gnosis.org
- Thomas the Apostle - Wikipedia, the free encyclopedia
- The Nag Hammadi copy of the Gospel of Thomas begins: "These are the secret sayings that the living Jesus spoke and Didymos Judas Thomas recorded.
- en.wikipedia.org
- The Story Of The Storytellers - The Gospel Of Thomas | From Jesus
- This book opens with the lines, "These are the secret words which the living Jesus spoke, and the twin, Didymos Judas Thomas wrote them down." Then there ...
- www.pbs.org
- tradition - Who was Thomas's twin? - Christianity Stack Exchange
- Mar 11, 2013 The sainted apostle "Doubting" Thomas is called Didymus, the Twin. ... written in the early third century, identifies him as "Judas Thomas", and ...
- christianity.stackexchange.com
Great care has been taken to prepare the information on this page. Elements of the content come from factual and lexical knowledge databases, realmagick.com library and third-party sources. We appreciate your suggestions and comments on further improvements of the site.
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Abel Sanchez: Ponamarev Likely Back on Lomachenko vs. Walters
World class trainer Abel Sanchez's deep stable of fighters includes the likes of Gennady Golovkin, Murat Gassiev, Andy Ruiz and new arrival Alex Saucedo. His stable also includes unbeaten welterweight...Read More
Errol Spence-Leonard Bundu Title Eliminator Ordered By IBF
It's likely that Errol Spence will have to move into the mandatory challenger position in order to force the hand of reigning welterweight titlist Kell Brook. The trick is finding a willing opponent t...Read More
Errol Spence: Vargas Will Need KO or Beat Brook's Ass To Win in UK
Errol Spence Jr. (20-0, 17 KOs) is still waiting for the situation to play out with the IBF. The sanctioning body ordered him to Konstantin Ponomarev (30-0, 13 KOs) in an IBF final eliminator. ...Read More
Hearn: No Way Does Spence's Team Put Him in With Brook Just Yet
Last weekend Errol Spence (20-0, 17KOs) created a tidal wave in the welterweight division after he demolished durable former champion Chris Algieri in five rounds. ...Read More
Errol Spence-Konstantin Ponomarev IBF Final Eliminator Ordered
Errol Spence (20-0, 17KOs) will have one final hurdle to overcome before he can secure a mandatory world title shot. The International Boxing Federation has ordered Spence and Konstantin Ponomarev (30...Read More
Pacquiao-Bradley Trilogy LIVE Results From Las Vegas
In an intriguing battle of undefeated welterweights with big implications, Konstantin Ponamarev (30-0) defeated Brad Solomon (26-1) via split decision in a bout he appeared to dominate. The North Ame...Read More
Pacquiao-Bradley: Ponomarev Wants To Go To War With Solomon
Russian welterweight contender Konstantin Ponomarev (29-0, 13KOs) detailed his recent sparring session against a former three division world champion, 44-year old Shane Mosley. ...Read More
Konstantin Ponomarev-Brad Solomon in Play for Pacquiao-Bradley
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Sanchez wants Ponamarev to get 4-5 more fights before title shot
23-year-old welterweight prospect Konstanin Ponomarev has already amassed a 29-0 record. That is quite impressive for a fighter of that age. With that being said, the Ural Promotions and Top Rank c...Read More
Ramirez-Khatchikian Off-TV Results: Ponomarev Wins
In the opening bout of the night, undefeated super featherweight prospect Toka Khan Clary (17-0) kept his perfect record intact with a victory over Sergio Lopez (18-11-1). Clary dropped Lopez in the ...Read More
Ponomarev Finds Added Motivation From Golovkin
There is nothing like having a comfort zone. For welterweight Konstantin Ponomarev, that comfort zone was fighting in front of familiar faces in his home country of Russia. But fighting for bigger ...Read More
Photos/Weights: Ramirez 167.5, Khatchikian 167
Gilberto Ramirez puts not only his unbeaten record but also a mandatory title shot on the line as he faces Netherland's Gevorg Khatchikian this Friday evening at The Cosmopolitan in Las Vegas. Both...Read More
Konstantin Ponomarev Now Set To Face Ramses Agaton
It's finally fight week for Konstantin Ponomarev. The unbeaten 23-year old from Russia has been waiting more than six months to hear those words, a bit of a downer since he's coming off of the big...Read More
Ponomarev's Return Pushed To Ramirez TruTV Bill
23-year-old Russian welterweight contender Konstantin Ponomarev (28-0, 13KOs) has received a new date for is next ring return. He was scheduled to come back on November 7th at the Thomas & Mack Center...Read More
Bradley-Rios: Konstantin Ponomarev Aims To Impress
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Gassiev, Ponomarev Open Camp With Abel Sanchez
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Ray Beltran Demolishes Ao, Ponomarev Shocks Zewski
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Photos/Weights: Ao is Ready, Beltran is Overweight
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Top Rank/TruTV Debut Card Ticket Info For 5/1 Card
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Beltran-Ao, Zewski-Ponomarev TruTV Card Announced
LAS VEGAS, NV. – The Chelsea inside the Cosmopolitan of Las Vegas welcomes an exciting evening of boxing featuring former two-division world champion TAKAHIRO AO who returns to the ring in his 2015 de...Read More


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For the Makers Inc.
Here is a brilliant idea for the Pinterest age.
I don't know about you, but I am routinely inundated with ideas for tempting do-it-yourself projects. Let's just take a quick peek at my Pinterest page. Yep, there's a project to turn a small toddler T-shirt into a dress, to turn an old dresser into a "signature" piece of furniture, to turn old furniture into garden ornaments, or turn chipped teacups into bird feeders. And that's not to mention the endless macrame, quilting, painting, antiquing, jewelry-making and paper-making efforts that look so simple and fun. »Read Full Blog Post
Take your best shot with Bronze Fonz
"Happy Days" are indeed here again! To celebrate the Bronze Fonz' fifth birthday Monday, you can share a photo in our reader gallery by using the submission form or by emailing your photo, with the caption in the subject line, to firstname.lastname@example.com. »Read Full Article
Harley-Davidson 110th Anniversary street parties to feature Sick Puppies, Puddle of Mudd
If 60 acts — including Toby Keith, Aerosmith and Kid Rock — at the Summerfest grounds weren't enough entertainment for you for Harley-Davidson's 110th Anniversary Celebration, the Milwaukee motorcycle company is bringing in even more bands for free street parties Labor Day weekend, including Sick Puppies and Puddle of Mudd.
The first street party takes place from 4 to 11:30 p.m. Aug. 30 on E. North Ave. between Prospect and Oakland avenues, featuring blues rock group Jonathan Tyler and the Northern Lights at 7:30 p.m., and Australian hard-rock trio Sick Puppies at 9:30 p.m. »Read Full Article(18)
Send us your Lebowski Fest photos
Were you at Lebowski Fest this weekend? Submit your best pictures of White Russians, "The Dude" impersonations and other costumes in honor of the cult-classic film, "The Big Lebowski," to be featured in our reader photo gallery. »Read Full Article
John Hodgman, Eugene Mirman, Kristen Schaal team up for Pabst Theater comedy show
It's a triple comic threat at the Pabst Theater Oct. 18 when three comedians with strong cult followings--John Hodgman, Eugene Mirman and Kristen Schaal--perform stand-up sets on the same stage.
The tour, dubbed "MirmanHodgmanSchaal Sandwich to Go," is only hitting six cities. »Read Full Article
APT closing summer strong with 'All My Sons,' 'Cleopatra'
Spring Green — On a sleepy summer Sunday in 1947, 61-year-old Joe Keller sits in his backyard, smoking a pipe while reading the paper. In the background, a dog barks and children play; church bells chime and insects buzz. The grass is lush and green, and the future seems as bright as the sky, which we're told doesn't have a cloud.
That's how things begin in the outstanding American Players Theatre production of Arthur Miller's "All My Sons," which director William Brown has set in a seemingly idyllic "Wisconsin neighborhood" resembling thousands like it throughout postwar America. »Read Full Article
Tedeschi Trucks Band, Dave Mason among new concert announcements
Blues supergroup the Tedeschi Trucks Band, featuring husband-and-wife team Susan Tedeschi and Derek Trucks, will return to the Riverside Theater Oct. 16. The band won the Best Blues Album Grammy with 2011 debut "Revelator," and is dropping its second studio effort, "Made Up Mind," Tuesday.
Tickets are $39.50 to $65 and are available for purchase beginning at noon Friday at the box office (116 W. Wisconsin Ave.), the Pabst Theater box office (144 E. Wells St.), by calling (414) 286-3663 or visiting pabsttheater.org. »Read Full Article
Hear Volcano Choir's album now on NPR
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On Tap for Today: Brewers, nature and Florida swamp music
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realmagick.com The shrine of knowledge.
Pitch Lake
A selection of articles related to pitch lake.
Original articles from our library related to the Pitch Lake. See Table of Contents for further available material (downloadable resources) on Pitch Lake.
- The Lady of the Lake
- This is simply a conflation of all the multitudinous lake, river, and water spirits so prevalent in Celtic mythology. Nevertheless, common threads do appear; one of the best documented is that of relic-guardian, holder of the sacred sword Excalibur, who gives...
Deities & Heros >> Celtic, Welsh, Irish & Brittish
- Thoughts on the Soul and Reincarnation
- Life is an effort to re-member one's connections to the all-knowing soul, which we have chosen to forget so that we may fully appriciate that knowlege by experiencing what it is like to not have that knowlege. We do not remember our choosing this...
Spirits >> The Soul
- King James Bible: Numbers, Chapter 2
- Chapter 2 2:1 And the LORD spake unto Moses and unto Aaron, saying, 2:2 Every man of the children of Israel shall pitch by his own standard, with the ensign of their father's house: far off about the tabernacle of the congregation shall they pitch. 2:3 And on...
Old Testament >> Numbers
- King James Bible: Numbers, Chapter 3
- Chapter 3 3:1 These also are the generations of Aaron and Moses in the day that the LORD spake with Moses in mount Sinai. 3:2 And these are the names of the sons of Aaron; Nadab the firstborn, and Abihu, Eleazar, and Ithamar. 3:3 These are the names of the...
Old Testament >> Numbers
- A Treatise on Astral Projection, Part 3: The Rope Techniques
- A key ingredient to the new projection techniques is an invisible, imaginary ROPE hanging from your ceiling. This ROPE will be used to exert dynamic pressure at a single point on your astral body to force its separation from the physical. The ROPE technique...
Parapsychology >> Astral Projection
- King James Bible: Revelation, Chapter 20
- Chapter 20 20:1 And I saw an angel come down from heaven, having the key of the bottomless pit and a great chain in his hand. 20:2 And he laid hold on the dragon, that old serpent, which is the Devil, and Satan, and bound him a thousand years, 20:3 And cast...
New Testament >> Revelation
- Everyday Water
- When you think of "Water" what comes to mind? A tranquil lake, gentle rain, or raging sea? Whether magical element or just a simple cup of tea, water can be a very special part of your life. Water Spirits Every body of water is an entity. Each lake,...
The Elements >> Water
Pitch Lake is described in multiple online sources, as addition to our editors' articles, see section below for printable documents, Pitch Lake books and related discussion.
Suggested News Resources
- The snow is off the beach, but what's left isn't pretty
- In its place, eroded banks and all of the stuff the lake spewed up during the howling storms of winter. ...
- Cuba and the United States Play 'Béisbol' Diplomacy
- The media image of Obama tossing the first pitch—if his security detail permits him to go on the field—will send a clear message to sports aficionados from Middle America that our common ground with Cuba includes the baseball diamond.
- New midfielder Antonio Nocerino fits easily with Orlando City
- It doesn't hurt that Nocerino already has found Italian restaurants in town he likes, Terramia Brick Oven Pizza in Lake Mary – whose owner, like Nocerino, is from Naples, Italy – and Armando's in Winter Park.
- Happ sharp in 2nd start, helping Blue Jays beat Rays 5-1
- Goins scored on the first wild pitch of the inning, and Toronto added another run when another pitch got away from Archer after he walked Junior Lake to re-load the bases.
- Flash funding highlights online philanthropy in class
- The celebrity givers in Thursday's campaign hope to inspire others to pitch in and get all the pending projects funded. Google co-founder Sergey Brin and WhatsApp co-founder Brian Acton will match $3.2 million in citizen donations.
Great care has been taken to prepare the information on this page. Elements of the content come from factual and lexical knowledge databases, realmagick.com library and third-party sources. We appreciate your suggestions and comments on further improvements of the site.
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The home inventory is a difficult process where you have to describe and write down every detail about every item in your home. This is extremely difficult, because everybody owns a lot of things and it's really hard to keep track of all of them. After all, things break, get lost, get damaged and are being thrown away every single day, so it's hard for some people to remember what they have at the moment. But the inventory is created exactly to help these people know what they have in every moment. But in order to conduct the inventory correctly, we need certain tools. Without them nothing would be possible. Let's check which are the best home inventory tools. Read the rest of The Best Home Inventory Tools >>
One particular invention in the end of 19th century, prove to be a...
The chickweed is a famous fast-growing plant, which is considered perennial herb. You...
If you have kids or you love chewing gum, this article will help...
The home inventory is a difficult process where you have to describe and...
The Best Home Inventory Tools (Jane Wilson posted on March 8th, 2013 )
COMMENTS: No Comments »
TAGS:home improvement, home inventory, inventory report, photography, property, rental property, tools
CATEGORIES: Home Improvement
Top 3 Easy Steps to Clean Your Mirror (Joanna Johnson posted on February 19th, 2013 )
Mirrors have a great role in our everyday life. Almost every one of us has a mirror in his house. Mirrors are the most important tool for our appearance. Without them we won't ever care about how we look and this would be really terrible. They help us notice the little changes our body is having. That's why we also have to take care of them, too.
Don't you hate it when you try to take a look at yourself in the mirror and it is all foggy and you can't see a thing? Mirrors are quite sensitive to moisture and dirt due to their glossy surface and structure. In order to keep your mirrors clean and shiny, you have to occasionally take care of them and that's why we made for you these top 3 easy steps to clean your mirror.
Read the rest of Top 3 Easy Steps to Clean Your Mirror >>
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TAGS:cleaning mirror, mirror fog, polishing
CATEGORIES: Home Improvement
How to Make More Time (smithmartin900 posted on February 18th, 2013 )
Finding the time to do all the mandatory things in one's day is quite a challenge, finding the time for all the fun and enjoyable things is almost impossible. Usually the stress, the hassle and worries of everyday living can get the better of people and leave them struggling for that extra bit of time to get everything done. Read the rest of How to Make More Time >>
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TAGS:being more productive, how to, more time
CATEGORIES: Self Improvement
How to Help Your Cat Adjust After Moving (Jane Wilson posted on February 14th, 2013 )
Moving to a new property can be really difficult and stressful for everybody. After all, you'll have to pack and move everything you own to a new place and then unpack it again. But the moving experience can be even more difficult when there's a cat involved in the process. Even if you love your cat very much, you're probably going to hate it when you move to the new apartment. This is because the cats usually get attached to the place they live in and it takes a lot of time until they get used to the new one. But there are a few simple tips on how to help your cat adjust after moving that can be very useful in this occasion.
Keep everything closed. When you finally move in to the new place, you should remember to keep all the doors and windows closed for the first few weeks. This is because your cat may decide that it wants to go back to the old place and try to escape from the new property. This is definitely something you don't want to happen, because it will be really difficult to find it afterwards. So, make sure that your cat is safely closed within the new apartment until it forgets about the old one. Read the rest of How to Help Your Cat Adjust After Moving >>
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TAGS:apartment, cats, help, home improvement, moving house, pets, property
CATEGORIES: Moving house