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Positive Control (positive + control)
Kinds of Positive Control Terms modified by Positive Control Selected AbstractsA laboratory assessment of coronal bacterial leakage in root canals filled with new and conventional sealersINTERNATIONAL ENDODONTIC JOURNAL, Issue 4 2009A. U. Eldeniz Abstract Aim, To evaluate the resistance to ex vivo bacterial leakage over a 40-day period of root canal fillings with five new root canal sealers: RC Sealer, Epiphany, EndoREZ, GuttaFlow and Acroseal, compared with Apexit, AH Plus and RoekoSeal. Methodology, One hundred and forty-four single rooted human teeth were divided randomly into eight test (n = 15) and two control groups (n = 12). The root canals were filled using a single cone technique with gutta-percha except in the Epiphany and EndoREZ groups. These were filled with Resilon and resin-coated gutta-percha, respectively. The gutta-percha surface of the GuttaFlow group was coated with an experimental primer prior to filling. Positive controls were filled with gutta-percha without sealer and tested with bacteria, whereas negative controls were sealed with wax to test the seal between the chambers. Filled roots were incorporated in a split chamber model system using Streptococcus mutans as a microbial marker. Leakage was assessed for turbidity of the broth in the lower chamber every day for 40 days. Survival analysis was performed using the Kaplan,Meier product limit method and event times were compared using the Log-rank test (, = 0.05). Results, Epiphany, GuttaFlow with test primer and Apexit prevented leakage significantly better than AH Plus, RC Sealer, RoekoSeal, EndoREZ and Acroseal (P < 0.05). None of the specimens in the AH Plus, RC Sealer, RoekoSeal and EndoREZ groups resisted bacterial penetration for 40 days. Conclusion, The new sealers, Epiphany and GuttaFlow with primer, along with Apexit, showed better resistance to bacterial penetration than the other new or traditional sealers tested. [source] Evaluation of root-end cavity preparation using ultrasonic retrotipsINTERNATIONAL ENDODONTIC JOURNAL, Issue 9 2003H. Ishikawa Abstract Aim, To evaluate and compare the efficiency of root-end preparations using ultrasonic retrotips coated with diamond and zirconium nitride. Methodology, Eighty-five extracted single-rooted teeth were root filled, and then resected 3 mm from their apices. Root-end cavities were prepared with KiS (zirconium nitride-coated retrotip), CT-5 (stainless steel tip) or diamond-coated (DC) ultrasonic retrotips, and 10 teeth served as controls. Thirty teeth were used for evaluation of the time required to prepare the root-end cavity, the number of microcracks produced on the resected surface and the number of dentinal tubule openings on the root-canal wall using scanning electron microscope (SEM) images. A further 55 teeth were used for evaluation of dye penetration following filling of the root-end cavities with Super EBA. The degree of dye penetration in millimetres was measured under the microscope after 7 days of immersion in India ink. Statistical analyses were performed using the one-way anova and Scheffe's F -test as the post hoc test. Results, There was no significant difference in the number of microcracks and dentinal tubule openings present in the root apices prepared by the three retrotips. The time required for root-end cavity preparation using the DC retrotip was significantly less than that using the other groups (P < 0.01). Positive controls showed dye penetration throughout the length of the root-end cavity, and negative controls showed no dye penetration. There was no significant difference between the three experimental groups in dye penetration. Conclusions, In this laboratory study, the time required to prepare root-end cavities using KiS retrotips was the same as that using CT-5 retrotips, and longer than that using DC retrotips. There was no significant difference in the number of microcracks or dye penetration between the three kinds of retrotips. [source] Evaluation of genotoxic effects in human leukocytes after in vitro exposure to 1950 MHz UMTS radiofrequency fieldBIOELECTROMAGNETICS, Issue 3 2008O. Zeni Abstract In the present study the third generation wireless technology of the Universal Mobile Telecommunication System (UMTS) signal was investigated for the induction of genotoxic effects in human leukocytes. Peripheral blood from six healthy donors was used and, for each donor, intermittent exposures (6 min RF on, 2 h RF off) at the frequency of 1950 MHz were conducted at a specific absorption rate of 2.2 W/kg. The exposures were performed in a transverse electro magnetic (TEM) cell hosted in an incubator under strictly controlled conditions of temperature and dosimetry. Following long duration intermittent RF exposures (from 24 to 68 h) in different stages of the cell cycle, micronucleus formation was evaluated by applying the cytokinesis block micronucleus assay, which also provides information on cell division kinetics. Primary DNA damage (strand breaks/alkali labile sites) was also investigated following 24 h of intermittent RF exposures, by applying the alkaline single cell gel electrophoresis (SCG)/comet assay. Positive controls were included by treating cell cultures with Mitomycin-C and methylmethanesulfonate for micronucleus and comet assays, respectively. The results obtained indicate that intermittent exposures of human lymphocytes in different stages of cell cycle do not induce either an increase in micronucleated cells, or change in cell cycle kinetics; moreover, 24 h intermittent exposures also fail to affect DNA structure of human leukocytes soon after the exposures, likely indicating that repairable DNA damage was not induced. Bioelectromagnetics 29:177,184, 2008. © 2007 Wiley-Liss, Inc. [source] Evaluation of genotoxic effects in human peripheral blood leukocytes following an acute in vitro exposure to 900 MHz radiofrequency fieldsBIOELECTROMAGNETICS, Issue 4 2005O. Zeni Abstract Human peripheral blood leukocytes from healthy volunteers have been employed to investigate the induction of genotoxic effects following 2 h exposure to 900 MHz radiofrequency radiation. The GSM signal has been studied at specific absorption rates (SAR) of 0.3 and 1 W/kg. The exposures were carried out in a waveguide system under strictly controlled conditions of both dosimetry and temperature. The same temperature conditions (37.0,±,0.1 °C) were realized in a second waveguide, employed to perform sham exposures. The induction of DNA damage was evaluated in leukocytes by applying the alkaline single cell gel electrophoresis (SCGE)/comet assay, while structural chromosome aberrations and sister chromatid exchanges were evaluated in lymphocytes stimulated with phytohemagglutinin. Alterations in kinetics of cell proliferation were determined by calculating the mitotic index. Positive controls were also provided by using methyl methanesulfonate (MMS) for comet assay and mitomycin-C (MMC), for chromosome aberration, or sister chromatid exchange tests. No statistically significant differences were detected in exposed samples in comparison with sham exposed ones for all the parameters investigated. On the contrary, the positive controls gave a statistically significant increase in DNA damage in all cases, as expected. Thus the results obtained in our experimental conditions do not support the hypothesis that 900 MHz radiofrequency field exposure induces DNA damage in human peripheral blood leukocytes in this range of SAR. Bioelectromagnetics 26:258,265, 2005. © 2005 Wiley-Liss, Inc. [source] Nonanoic acid , an experimental irritantCONTACT DERMATITIS, Issue 3 2003Jan E. Wahlberg Irritant contact dermatitis is defined as a non-immunological skin reaction following exposure to various chemical, mechanical and physical factors. It is known that the skin response to irritants depends on the irritant applied and differs between chemically different irritants. Sodium lauryl sulfate (SLS) is an anionic detergent and the most frequently used substance in experimental irritant contact dermatitis. In 1980, it was suggested that nonanoic acid (NNA) could be used as a positive control when patch testing. Since then, NNA has been used as an experimental irritant in several studies and has been used as a chemically different substance compared to SLS. The present article presents a review of the application of NNA in studies on skin irritancy and experimental irritant contact dermatitis. [source] Viability of fibroblasts in a novel probiotic storage mediaDENTAL TRAUMATOLOGY, Issue 5 2010E Ēaglar The aim of the present in vitro study was to evaluate the number of viable PDL cells of avulsed teeth treated by Hank's Balanced Salt Solutions (HBSS), saline, a novel probiotic solution and milk. Thirty-six freshly extracted single-rooted human teeth with closed apices were divided into one of the four experimental groups and two control groups (N = 6 each). The positive and negative controls corresponded to 0 min and an 8-h dry time respectively. Following extraction, the coronal 3 mm of PDL tissue was scraped with a #15 scalpel to remove cells that might have been damaged. The experimental teeth were dried for 30 min followed by a 45 min immersion in one of the four experimental media. Each experimental tooth, after drying and soaking, was incubated for 30 min with a 2.5 ml solution of 0.2 mg ml,1 of collagenase CLS II and a 2.4 mg ml,1 solution of dispase grade II in phosphate buffer saline (PBS). The cells were then labelled with 0.4% Trypan blue for determination of viability. The teeth stored in positive control demonstrated the highest number of viable PDL cells followed in rank order by HBSS, saline, Lactobacillus reuteri solution and milk. There was no significant difference in the number of viable PDL cells between HBSS, milk, L. reuteri solution and saline. Within the parameters of this study, it appears that probiotic may be able to maintain PDL cell viability as HBSS, milk, or saline. [source] Effect of temperature and storage media on human periodontal ligament fibroblast viabilityDENTAL TRAUMATOLOGY, Issue 3 2010Beatriz Dulcineia Mendes Souza The purpose of this study was to compare the effectiveness of several storage media to preserve cultured periodontal ligament fibroblasts (PDLF) under different temperatures. The media tested were: sterile Hank's balanced salt solution (sHBSS), non-sterile HBSS (nHBSS), skimmed milk, Save-A-Tooth®, Minimum Essential Medium (MEM) and water (negative control). MEM at 37°C was used as positive control. PDLF were obtained from explants of extracted healthy human teeth. Plates containing confluent PDLF were soaked in the various media for 3, 6, 24, 48 and 72 h at 37°C and 20°C. After incubation, viability of the cells was determined using the tetrazolium salt-based colorimetric (MTT) assay and the Trypan Blue exclusion test after 6, 24, 48 and 72 h of incubation at 20°C. The results were analyzed statistically using Kruskal,Wallis, Scheffé and Mann,Whitney (, = 5%) tests. Results from the MTT assay at 37°C and 20°C showed that skimmed milk was the best storage medium for up to 24 and 48 h, respectively, followed by nHBSS and sHBSS. Results from the Trypan Blue exclusion test showed that the best storage media were milk, sHBSS and nHBSS, with no statistical differences, for any time period. The Save-A-Tooth® had a detrimental effect on cells after 24 h. The influence of temperature on the effectiveness of the storage media tested showed at 20°C a decreasing order of efficacy as follows: milk > sHBSS and nHBSS > MEM > Save-A-Tooth® > water while at 37°C it was: MEM > nHBSS > milk > sHBSS > Save-A-Tooth® > water. In conclusion, incubation temperature altered the effectiveness of the storage media and skimmed milk at 20°C was better than HBSS in maintaining PDLF viability. [source] Comparative in vitro study of the sealing efficiency of white vs grey ProRoot mineral trioxide aggregate formulas as apical barriersDENTAL TRAUMATOLOGY, Issue 2 2008Spyridon Stefopoulos Recently conventional grey MTA has been replaced by a new white MTA formula. The aim of this study was to compare the root canal adaptation of white MTA to that of grey MTA when used as an apical barrier in teeth with open apices. We also examined whether a previous calcium hydroxide intracanal medication affects MTA's sealing ability and investigated the ability to remove calcium hydroxide from the root canal walls. Forty-nine teeth were prepared in a manner to simulate a divergent open apex of immature teeth. Four teeth were used in a preliminary experiment to demonstrate the inefficacy of calcium hydroxide removal from the canal walls in teeth with open apices. Four groups of 10 teeth each were created: groups A and B were treated with calcium hydroxide intracanal medication and then received an apical plug of grey and white MTA respectively. Groups C and D received an apical plug of grey and white MTA respectively without previous intracanal medication. Four teeth served as negative and one as a positive control. The marginal adaptation and sealing ability of the apical barrier were tested by means of a dye tracer (basic fuchsine) after longitudinal sectioning. It was found that MTA apical barrier resisted displacement during gutta-percha condensation. Calcium hydroxide pretreatment, adversely affected white MTA sealing ability (P < 0.05). [source] Pulp revascularization of replanted immature dog teeth after treatment with minocycline and doxycycline assessed by laser Doppler flowmetry, radiography, and histologyDENTAL TRAUMATOLOGY, Issue 2 2004Alessandra Luisa de Souza Ritter Abstract,,, This study investigated the effect of topical antibiotic treatment on pulp revascularization in replanted teeth. Thirty-four immature teeth were selected from three young dogs. Baseline radiographs and laser Doppler flowmetry (LDF) readings were obtained. Specimens were randomly divided into four groups: Thirty-eight teeth were extracted, kept dry for 5 min, and either (Group 1) covered with minocycline mixture (G1, n = 11), (Group 2) soaked in doxycycline (G2, n = 11), or (Group 3) soaked in saline (G3-negative control, n = 6), and replanted. Teeth in Group 4 were not extracted (positive control, n = 6). Postoperative radiographs and LDF readings were obtained for 2 months after replantation. After sacrifice, the jaws were collected and processed for light microscopy. Pre- and postreplantation LDF readings and radiographs, and histologic findings were analyzed to assess revascularization. Pulp revascularization occurred in 91% (G1), 73% (G2), and 33% (G3) of the specimens. In conclusion, minocycline facilitates pulp revascularization in replanted immature teeth after replantation. [source] Viability study of HL60 cells in contact with commonly used microchip materialsELECTROPHORESIS, Issue 24 2006Floor Wolbers Abstract This paper presents a study in which different commonly used microchip materials (silicon oxide, borosilicate glass, and PDMS) were analyzed for their effect on human promyelocytic leukemic (HL60) cells. Copper-coated silicon was analyzed for its toxicity and therefore served as a positive control. With quantitative PCR, the expression of the proliferation marker Cyclin D1 and the apoptosis marker tissue transglutaminase were measured. Flow cytometry was used to analyze the distribution through the different phases of the cell cycle (propidium iodide, PI) and the apoptotic cascade (Annexin V in combination with PI). All microchip materials, with the exception of Cu, appeared to be suitable for HL60 cells, showing a ratio apoptosis/proliferation (Rap) comparable to materials used in conventional cell culture (polystyrene). These results were confirmed with cell cycle analysis and apoptosis studies. Precoating the microchip material surfaces with serum favor the proliferation, as demonstrated by a lower Rap as compared to uncoated surfaces. The Cu-coated surface appeared to be toxic for HL60 cells, showing over 90% decreased viability within 24,h. From these results, it can be concluded that the chosen protocol is suitable for selection of the cell culture material, and that the most commonly used microchip materials are compatible with HL60 culturing. [source] Genotoxicity testing of the herbicide trifluralin and its commercial formulation Treflan using the piscine micronucleus testENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 6 2008Serpil Könen Abstract In this study, the genotoxic effects of a widely used herbicide, trifluralin, and its commercial formulation, Treflan, were evaluated using the micronucleus test in a commercially important fish species, Oreochromis niloticus (Nile Tilapia). Fish were exposed to 1, 5, and 10 ,g/L doses of trifluralin and Treflan for 3, 6, and 9 days under laboratory conditions. Ethylmethanesulfonate, at a single dose of 10 mg/L, was used as positive control. Micronuclei were evaluated on the peripheral erythrocytes. Both Treflan and trifluralin treatments significantly increased the micronucleus frequencies in peripheral erythrocytes of O. niloticus. Furthermore, the genotoxicity of the active ingredient, trifluralin, was observed to be higher than that of the commercial formulation Treflan. Our results indicate that herbicide trifluralin has genotoxic potential in fish. Environ. Mol. Mutagen., 2008. © 2008 Wiley-Liss, Inc. [source] Evaluation of river water genotoxicity using the piscine micronucleus testENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 6 2007Serap Ergene Abstract The Berdan River, which empties into the Mediterranean Sea on the east coast of Turkey, receives discharges of industrial and municipal waste. In the present study, the in vivo piscine micronucleus (MN) test was used to evaluate the genotoxicity of water samples collected from different locations along the Berdan River. Nile tilapia (Oreochromis niloticus) were exposed in the laboratory for 2, 4, and 6 days, and micronuclei were evaluated in peripheral blood erythrocytes, gill cells, and caudal fin epithelial cells. A single dose of 5 mg/L cyclophosphamide was used as a positive control. In addition to micronuclei, nuclear abnormalities (NAs), such as binucleated cells and blebbed, notched, and lobed nuclei, were assessed in the erythrocytes, and chemical analyses were carried out to determine the amount of heavy metals in the water samples. MN and NA frequencies were significantly elevated (up to 2- to 3-fold) in fish exposed to river water samples taken downstream of potential discharges, and the elevated responses in gill and fin cells were related to the concentration of heavy metals in the water. MN frequencies (expressed as micronucleated cells/1,000 cells), in both treated and untreated fish, were greatest in gill cells (range: 0.80,3.70), and generally lower in erythrocytes (range: 0.50,2.80), and fin cells (range: 0.45,1.70). The results of this study indicate that the Berdan River is contaminated with genotoxic pollutants and that the genotoxicity is related to the discharge of wastes into the river water. Environ. Mol. Mutagen., 2007. © 2007 Wiley-Liss, Inc. [source] In vivo exposure to microcystins induces DNA damage in the haemocytes of the zebra mussel, Dreissena polymorpha, as measured with the comet assayENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 1 2007Guillaume Juhel Abstract The Comet assay was used to investigate the potential of the biotoxin microcystin (MC) to induce DNA damage in the freshwater zebra mussel, Dreissena polymorpha. Mussels maintained in the laboratory were fed daily, over a 21-day period, with one of four strains of the cyanobacterium, Microcystis aeruginosa. Three of the strains produced different profiles of MC toxin, while the fourth strain did not produce MCs. The mussels were sampled at 0, 7, 14, and 21 days by withdrawing haemocytes from their adductor muscle. In addition, a positive control was performed by exposing a subsample of the mussels to water containing cadmium chloride (CdCl2). Cell viability, measured with the Fluorescein Diacetate/Ethidium Bromide test, indicated that the MC concentrations, to which the mussels were exposed, were not cytotoxic to the haemocytes. The Comet assay performed on the haemocytes indicated that exposure to CdCl2 produced a dose-responsive increase in DNA damage, demonstrating that mussel haemocytes were sensitive to DNA-damaging agents. DNA damage, measured as percentage tail DNA (%tDNA), was observed in mussels exposed to the three toxic Microcystis strains, but not in mussels exposed to the nontoxic strain. Toxin analysis of the cyanobacterial cultures confirmed that the three MC-producing strains exhibit different toxin profiles, with the two MC variants detected being MC-LF and MC-LR. Furthermore, the DNA damage that was observed appeared to be strain-specific, with high doses of MC-LF being associated with a higher level of genotoxicity than low concentrations of MC-LR. High levels of MC-LF also seemed to induce relatively more persistent DNA damage than small quantities of MC-LR. This study is the first to demonstrate that in vivo exposure to MC-producing strains of cyanobacteria induces DNA damage in the haemocytes of zebra mussels and confirms the sublethal toxicity of these toxins. Environ. Mol. Mutagen., 2007. © 2006 Wiley-Liss, Inc. [source] Aqueous exposure to 4-nonylphenol and 17,-estradiol increases stress sensitivity and disrupts ion regulatory ability of juvenile Atlantic salmonENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 7 2007Darren T. Lerner Abstract Population declines of wild Atlantic salmon have been attributed to an array of anthropogenic disturbances, including dams, commercial and recreational fishing, habitat loss, and pollution. Environmental contaminants in particular, can act as environmental stressors on fish, typically causing disruption of ion homeostasis due to their close association with the aquatic environment. To examine the effects of the xenoestrogen 4-nonylphenol (NP) or 17,-estradiol (E2) on stress sensitivity and ion regulation, we exposed juvenile Atlantic salmon continuously for 21 d to either 10 or 100 ,g/L NP (NP-L or NP-H), 2 ,g/L E2 (positive control), or vehicle control during the parr-smolt transformation in April. After treatment, fish were sampled in freshwater (FW), transferred to 30, seawater (SW) for 24 h, or subjected to a handling stress. Estradiol and NP-H increased plasma vitellogenin in males and females, and E2 increased gonadosomatic index only in males. In FW, E2 reduced sodium potassium,activated adenosine triphosphatase activity as well as plasma levels of growth hormone, insulin-like growth factor I, and triiodothyronine. Both E2 and NP-H reduced plasma sodium in FW and increased plasma chloride in SW. Plasma Cortisol levels pre- and poststressor were significantly elevated by all treatments relative to controls, but only E2 increased plasma glucose before and after the stressor. These results indicate that exposure of anadromous salmonids to environmental estrogens heightens sensitivity to external stressors, impairs ion regulation in both FW and SW, and disrupts endocrine pathways critical for smolt development. [source] In vitro and in vivo estrogenicity and toxicity of o -, m -, and p -dichlorobenzeneENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 2 2003Bram J. Versonnen Abstract The estrogenicity of o -, m -, and p -dichlorobenzene (DCB) was evaluated with a yeast estrogen screen (YES) and zebrafish (Danio rerio) vitellogenin (VTG) assays. With the YES, p -DCB and m -DCB were found to be estrogenic in a concentration-responsive manner. The relative potency measured with the YES (relative to 17,-estradiol) was 2.2 × 10,7 for p -DCB and 1.04 × 10,8 for m -DCB. Following acute toxicity tests with the zebrafish, plasma VTG production was measured to examine the in vivo estrogenic activity of the three compounds after a 14-d exposure. Adult zebrafish were exposed to different concentrations of o -, m - and p -DCB, ranging from 0.1 to 32 mg/L; ethynylestradiol ([EE2]; 5 ng/L, 10 ng/L, 50 ng/L, and 100 ng/L) was used as a positive control. After exposure, blood samples were taken and protein electrophoresis was performed to determine the relative VTG content. Gonadosomatic indices (GSI) and condition factors (CF) were also calculated. Elevated VTG levels and decreased female GSIs were found in fish exposed to ,5 ng EE2/L and in fish exposed to ,10 mg p -DCB/L. Low GSIs coincided with high levels of VTG in the blood of female zebrafish. This relation was not only found in fish exposed to EE2 but also in controls and fish exposed to DCB. Therefore, a direct or indirect effect of VTG on the GSI is suggested rather than a direct toxic effect of the tested compounds on the gonads. [source] Effects of handling on heat shock protein expression in rainbow trout (Oncorhynchus mykiss)ENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 3 2002Barbara Shayne Washburn Abstract As part of an effort to validate the use of heat shock proteins (HSPs) as biomarkers of exposure to and effects of contaminants, we evaluated the effect of two handling regimens on the induction of HSP 60 and 70 in rainbow trout (Oncorhynchus mykiss). Fish were acclimated to laboratory conditions for several weeks before the beginning of the experiment. Fish were then captured by net, placed in a cooler for 1 h while being transported in a truck, returned to their original tanks, then sacrificed 6 to 8 h later. Tricaine methane sulfonate (MS-222) was used during different phases of handling to reduce handling stress. Heat-stressed fish were included in the experiment as a positive control. Muscle, liver, gills, and heart were analyzed for HSP 60 and 70 by immunoblotting. We found no effect of any handling regimen on the induction of HSPs. These findings suggest that the capture and transport of fish for environmental monitoring purposes should not interfere with the use of stress proteins as biomarkers. [source] Effects of bisphenol A and tetrabromobisphenol A on sex organ development in quail and chicken embryosENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 12 2001Cecilia Berg Abstract The plastic monomere bisphenol A (BPA) and the flame retardant tetrabromobisphenol A (TBBPA) were examined for estrogen-like developmental effects on the reproductive organs in avian embryos. The synthetic estrogen diethylstilbestrol (DES) was used as a positive control. The test compounds were injected into the yolk of quail and chicken eggs early during incubation and the embryos were examined 2 d before anticipated hatching. At 200 ,g/g egg, BPA induced Müllerian duct (embryonic oviduct) malformation in female quail embryos and feminization of the left testis (ovotestis) in male chicken embryos. The estrogenic potency of BPA compared with DES was species and endpoint specific. Müllerian duct malformation was the most sensitive endpoint in quail embryos, whereas ovotestis formation was the most sensitive response in chicken embryos. Tetrabromobisphenol A caused high embryo mortality at 45 ,g/g egg in both species, but no estrogen-like effects were observed. Bisphenol A caused mortality only in chicken embryos at 67 and 200 ,g/g egg. To our knowledge, this is the first report on estrogen-like or embryolethal effects of BPA and TBBPA in birds. [source] Substances with estrogenic activity in effluents of sewage treatment plants in southwestern Germany.ENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 10 2001Abstract The proliferation test with human estrogen receptor-positive MCF-7 breast cancer cells (E-Screen assay) was applied for quantitative determination of total estrogenic activity in 24-h composite effluent samples from 16 municipal and two industrial sewage treatment plants (STPs) in the state of Baden-Württemberg, southwestern Germany. The estrogenic efficacy relative to the positive control, 17,-estradiol, was between 26 and 74% (median, 48%) for the 16 municipal STPs. Estradiol equivalent concentrations (EEQs) were between 0.2 and 7.8 ng/L (median, 1.6 ng/L) and, thereby, were lower than those found in a pilot study, which revealed EEQs of greater than 10 ng/L in the effluents of two other STPs. The EEQs in 14 of the 16 effluent samples were very similar (0.9,3.3 ng/L), indicating a rather constant input of estrogenic substances via STPs into rivers. Additional activated charcoal filtration turned out to be very efficient in further eliminating estrogenic activity from effluents. The EEQs of the E-Screen assay and those calculated from the results of extensive chemical analysis using the estradiol equivalency factors determined for 13 natural and synthetic estrogenic substances were comparable for most of the effluent samples. 17,-Estradiol, 17,-ethinylestradiol, and, to a lesser extent, estrone contributed to 90% or more of the EEQ value. [source] Heat Shock Protein-27 Is Upregulated in the Temporal Cortex of Patients with EpilepsyEPILEPSIA, Issue 12 2004Hans-J Bidmon Summary:,Purpose: Heat shock protein-27 (HSP-27) belongs to the group of small heat shock proteins that become induced in response to various pathologic conditions. HSP-27 has been shown to protect cells and subcellular structures, particularly mitochondria, and serves as a carrier for estradiol. It is a reliable marker for tissues affected by oxidative stress. Oxidative stress and related cellular defence mechanisms are currently thought to play a major role during experimentally induced epileptic neuropathology. We addressed the question whether HSP-27 becomes induced in the neocortex resected from patients with pharmacoresistant epilepsy. Methods: Human epileptic temporal neocortex was obtained during neurosurgery, and control tissue was obtained at autopsy from subjects without known neurologic diseases. The tissues were either frozen for Western blot analysis or fixed in Zamboni's fixative for the topographic detection of HSP-27 at the cellular level by means of immunohistochemistry. Results: HSP-27 was highly expressed in all epilepsy specimens and in the cortex of a patient who died in the final stage of multiple sclerosis (positive control), whereas only low amounts of HSP-27 were detectable in control brains. In epilepsy patients, HSP-27 was present in astrocytes and in the walls of blood vessels. The intracortical distribution patterns varied strongly among the epilepsy specimens. Conclusions: These results demonstrate that HSP-27 becomes induced in response to epileptic pathology. Although the functional aspects of HSP-27 induction during human epilepsy have yet to be elucidated, it can be concluded that HSP-27 is a marker for cortical regions in which a stress response has been caused by seizures. [source] Tauroursodeoxycholic acid mobilizes ,-PKC after uptake in human HepG2 hepatoma cellsEUROPEAN JOURNAL OF CLINICAL INVESTIGATION, Issue 6 2002Helena Glasova Abstract Background Tauroursodeoxycholic acid (TUDCA) may exert anticholestatic effects via Ca++ - and ,-protein kinase C (,-PKC)-dependent apical vesicular insertion of canalicular transporters in cholestatic hepatocytes (Hepatology 2001; 33: 1206,16). Tauroursodeoxycholic acid is mainly taken up into liver cells by Na+ -taurocholate cotransporting polypeptide (Ntcp). Tauroursodeoxycholic acid selectively translocates ,-PKC, a key mediator of regulated exocytosis, to hepatocellular membranes. It is unclear whether TUDCA exerts its effects on ,-PKC after carrier-mediated uptake into liver cells or by interaction with extracellular/membraneous structures. Materials and methods Human hepatoblastoma HepG2 cells lacking Ntcp were stably transfected with pcDNA3·1/Ntcp or sham-transfected with pcDNA3·1 [+]. Distribution of ,-PKC was studied using a Western blotting technique. Uptake of [3H]taurocholic acid (TCA) was determined radiochemically. Results [3H]taurocholic acid uptake was approximately 180-fold higher in Ntcp-transfected than in sham-transfected cells. Phorbol 12-myristate 13-acetate (1 µmol L,1; positive control) increased membrane binding of ,-PKC by 34% in Ntcp-transfected and by 37% in sham-transfected cells. Tauroursodeoxycholic acid (10 µmol L,1) increased membrane-associated ,-PKC by 19% in Ntcp-transfected, but not in sham-transfected cells (,13%). Taurocholic acid (10 µmol L,1) did not affect the distribution of ,-PKC. Conclusion Carrier-mediated uptake is a prerequisite for TUDCA-induced translocation of ,-PKC to hepatocellular membranes. [source] Neuropsychiatric disturbances in SLE are associated with antibodies against NMDA receptorsEUROPEAN JOURNAL OF NEUROLOGY, Issue 5 2005R. Omdal To determine whether neuropsychiatric manifestations in patients with systemic lupus erythematosus (SLE) are influenced by antibodies against the human N-methyl- d -aspartate (NMDA) receptor types NR2a or NR2b. A decapeptide was synthesized containing a sequence motif present in the extracellular ligand-binding domain of NMDA receptors NR2a and NR2b, bound by the monoclonal murine anti-DNA antibody R4A. In an ELISA with the murine monoclonal R4v as positive control, plasma samples of 57 patients with SLE were examined for the anti-peptide (anti-NR2) antibody after the patients had been subjected to comprehensive psychological and cognitive testing. Poor performance on the Visual Paired Associates test (immediate), the Grooved Pegboard test, as well as high scores on the Beck Depression Inventory, and scales D-2 (depression), Pd-4 (psychopathic deviate), Sc-8 (schizophrenia), and Ma-9 (hypomania) of the MMPI-2 were significantly associated with elevated levels of anti-NR2 antibodies. The findings in several domains indicate an association between anti-NR2 antibodies and depressed mood in addition to decreased short-time memory and learning. Antibodies to NMDA receptors thus may represent one of several mechanisms for cerebral dysfunction in patients with SLE. [source] N -methyl- d -aspartate-triggered neuronal death in organotypic hippocampal cultures is endocytic, autophagic and mediated by the c-Jun N-terminal kinase pathwayEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 3 2003Tiziana Borsello Abstract Acute excitotoxic neuronal death was studied in rat organotypic hippocampal slices exposed to 100 µmN -methyl- d -aspartate. Fulgurant death of pyramidal neurons occurred in the CA1 and CA3 regions and was already detectable within 2 h of the N-methyl- d -aspartate administration. Morphologically, the neuronal death was neither apoptotic nor necrotic but had the hallmarks of autophagic neuronal death, as shown by acid phosphatase histochemistry in both CA1 and CA3 and by electron microscopy in CA1. The dying neurons also manifested strong endocytosis of horseradish peroxidase or microperoxidase, occurring probably by a fluid phase mechanism, and followed, surprisingly, by nuclear entry. In addition to these autophagic and endocytic characteristics, there were indications that the c-Jun N-terminal kinase pathway was activated. Its target c-Jun was selectively phosphorylated in CA1, CA3 and the dentate gyrus and c-Fos, the transcription of which is under the positive control of c-Jun N-terminal kinase target Elk1, was selectively up-regulated in CA1 and CA3. All these effects, the neuronal death itself and the associated autophagy and endocytosis, were totally prevented by a cell-permeable inhibitor of the interaction between c-Jun N-terminal kinase and certain of its targets. These results show that pyramidal neurons undergoing excitotoxic death in this situation are autophagic and endocytic and that both the cell death and the associated autophagy and endocytosis are under the control of the c-Jun N-terminal kinase pathway. [source] Microcell-mediated transfer of chromosome 4 into HeLa cells suppresses telomerase activityGENES, CHROMOSOMES AND CANCER, Issue 2 2001Claudia Backsch Telomerase activity can be detected in most human cancers and immortal cell lines. In contrast, the lack of telomerase activity in normal diploid fibroblasts has been correlated with progressive reduction of telomere lengths to critically short sizes followed by the cessation of cell division and the onset of senescence. Several investigators have provided evidence for the localization of a telomerase suppressor gene on chromosome 3. The aim of our study was to determine whether other chromosomes are involved in telomerase repression. Beside human chromosome 3 (serving as positive control), chromosomes 4, 6, and 11 were introduced into HeLa cells via microcell-mediated chromosome transfer. Telomerase activity from different hybrid cell lysates was determined at an early time point after fusion using a Telomerase ELISA kit. Strong repression of telomerase activity was only found in a subset of HeLa hybrids in which chromosome 3 or chromosome 4 had been introduced. Telomerase suppression induced by chromosome 3 or 4 transfer was paralleled by a high frequency (30% or 43%, respectively) of a senescent-like phenotype. Chromosomes 6 and 11, the functional loss of which is also implicated in cervical cancer, had no effect. These results indicate that normal human chromosomes 3 and 4 carry a gene or genes that suppress telomerase activity and induce cellular senescence in HeLa cells.©2001 Wiley-Liss, Inc. [source] Neural cell adhesion molecule expression: No correlation with perineural invasion in cutaneous squamous cell carcinoma of the head and neckHEAD & NECK: JOURNAL FOR THE SCIENCES & SPECIALTIES OF THE HEAD AND NECK, Issue 6 2009C. Arturo Solares MD Abstract Background. Perineural invasion (PNI) in cutaneous squamous cell carcinoma of the head and neck (CSCCHN) is associated with decreased survival, particularly in patients with clinical signs of cranial nerve involvement. There is evidence to indicate that neural cell adhesion molecule (N-CAM) confers capability of PNI. We analyzed our own patient population to determine if N-CAM predicted clinical PNI in CSCCHN. Methods. Tissue from patients with CSCCHN and clinical PNI, who underwent surgery between 1998 and 2005, was immunostained for N-CAM. In addition, non-PNI CSCCHN and normal nerve sections were also stained. A section of neuroendocrine tumor was included in each slide as a positive control. In addition, most of the sections also had an "inbuilt control" in the CD56 positive natural killer T cells that formed part of the inflammatory reaction to the tumors. Results. Tissue was available from 14 patients with CSCCHN and clinical PNI. The analysis was carried out in 14 patients without PNI and 4 normal nerves. N-CAM was not expressed in any of our PNI CSCCHN specimens or non-PNI controls. It was strongly expressed in the neuroendocrine tumors and positive in-built controls, as well as in normal nerve tissue. Conclusion. N-CAM expression did not predict neurotropism in our patient population. Additional studies are required to identify the cell surface markers expressed by CSCCHN which confer neurotropism capabilities. © 2009 Wiley Periodicals, Inc. Head Neck, 2009 [source] Benzodiazepine Alkaloids from Marine-Derived Endophytic Fungus Aspergillus ochraceusHELVETICA CHIMICA ACTA, Issue 7 2009Chuan-Ming Cui Abstract A new fungus-derived benzodiazepine analogue, 2-hydroxycircumdatin C (1), and a compound which has been isolated from a natural resource for the first time, but has been previously synthesized, namely (11aS)-2,3-dihydro-7-methoxy-1H -pyrrolo[2,1- c][1,4]benzodiazepine-5,11(10H,11aH)-dione (2), along with five structurally related known alkaloids (3,7), were isolated from Aspergillus ochraceus, an endophytic fungus derived from the marine brown alga Sargassum kjellmanianum. Their structures were established on the basis of spectroscopic methods. The absolute configuration of 1 was determined through CD evidence. Compound 1 displayed significant DPPH radical-scavenging activity with an IC50 value of 9.9,,M, which is 8.9-fold more potent than that of butylated hydroxytoluene (BHT), a well-known synthetic positive control. [source] The effect of irrigation time, root morphology and dentine thickness on tooth surface strain when using 5% sodium hypochlorite and 17% EDTAINTERNATIONAL ENDODONTIC JOURNAL, Issue 3 2010O. E. Sobhani Sobhani OE, Gulabivala K, Knowles JC, Ng Y-L. The effect of irrigation time, root morphology and dentine thickness on tooth surface strain when using 5% sodium hypochlorite and 17% EDTA. International Endodontic Journal, 43, 190,199, 2010. Abstract Aim, To evaluate the effect of irrigation with 5% sodium hypochlorite (NaOCl) alone and in conjunction with 17% ethylenediaminetetraacetic acid (EDTA) on tooth surface strain (TSS) and to analyse the influence of irrigation time, root morphology and dentine thickness. Methodology, Thirty-six single-rooted pre-molars with single canals had their crown and enamel reduced and root canals prepared using a standardized protocol. Teeth were grouped according to anatomical criteria and randomly distributed to experimental irrigation groups: (A) saline (negative control); (B) 5% NaOCl (positive control); (C) 5% NaOCl alternated with 17% EDTA. TSS was measured using electrical strain gauges bonded to the cervico-proximal part of the tooth. Teeth, mounted in clear acrylic resin placed in a universal testing machine, were subjected to nine consecutive 10-min irrigation periods followed by non-destructive occlusal loading to record TSS. Statistical analysis was carried out using two-way hierarchical anova and post hoc multiple comparisons. Results, Two groups showed an increase in TSS from the baseline (initial 10-min irrigation with saline). Group A showed a negligible reduction of 1.2% (343,339 ,,), which was not statistically significant (P = 0.7). Group B showed a highly significant (P = 0.001) increase in TSS by 53.7% (178,253 ,,), and group C showed a significant (P = 0.02) increase in TSS by 17.4% (163,192 ,,). The rate of change in TSS was significantly different between test groups. The length of the tooth (P = 0.04) as well as the mesio-distal (P = 0.05) width had significant effects on TSS. Conclusions, Irrigation with 5% with or without 17% EDTA increased TSS. The increase was significantly greater with 5% NaOCl alone than with 5% NaOCl alternated with 17% EDTA in contrast to previous findings with longer duration of irrigant exposure. Tooth length and mesio-distal root width significantly contributed to the increase in TSS. [source] Prostaglandin E2 production and viability of cells cultured in contact with freshly mixed endodontic materialsINTERNATIONAL ENDODONTIC JOURNAL, Issue 5 2006K. K. Melegari Abstract Aim, To determine whether commonly used endodontic sealers could either induce or increase the release of prostaglandin E2 (PGE2) when in contact with cell types found in the periapical tissues. Methodology, Freshly mixed samples of Roth 801 sealer, Sealapex® and ProRoot® mineral trioxide aggregate (MTA) were placed in contact with cultured macrophages and fibroblasts for 24 h. The supernatant from the cultures was assayed for PGE2 using enzyme-linked immunosorbent assay. Cell viability counts were made. As a positive control, similar cultures were also exposed to lipopolysaccharide and the supernatant analysed for PGE2. Data were compared by anova. Results, The three materials examined in these experiments did not stimulate increased PGE2 release from either of the cell lines. In control cultures, lipolysaccharide increased PGE2 release from macrophages but not from fibroblasts. Viability counts revealed that, whilst Roth 801 sealer caused some cell death in both fibroblasts and macrophages, Sealapex® led to cell death only in the macrophage cultures. ProRoot® MTA did not lead to statistically significant cell death in either culture. Conclusions, Under 24-h culture conditions, the three freshly mixed test materials did not increase directly either production or release of PGE2 from either macrophages or gingival fibroblasts. Roth 801 decreased cell viability counts for both fibroblasts and macrophages. Sealapex® decreases macrophage viability. ProRoot® MTA did not affect viability in either cell line. [source] In vitro antimicrobial effect of chlorhexidine-impregnated gutta percha points on Enterococcus faecalisINTERNATIONAL ENDODONTIC JOURNAL, Issue 2 2004J. N. Lui Abstract Aim, To evaluate the in vitro antimicrobial effect of chlorhexidine-impregnated gutta percha points, Roeko activ point (Roeko, Langenau, Germany) on Enterococcus faecalis. Methodology, Human maxillary premolar roots were prepared with .04 rotary ProFile instruments to a master apical file size 40, autoclave-sterilized and then infected with E. faecalis (ATCC 29212) for 3 weeks. Baseline controls were carried out verifying negligible effects of plain gutta percha cones on E. faecalis. Subsequent to intracanal placement of calcium hydroxide, ,activ points' or saline (positive control) and the 2-week incubation in 54 root specimens, dentine sampling at depths of 100 and 250 µm was carried out using .04 rotary ProFile instruments at sizes 60 and 90 to assess the quantity of bacteria present. Inactivating agents were used prior to sampling and the colony-forming units (CFU) of E. faecalis were then plate-counted after culturing. Statistical analysis was completed using the paired t -test. Results, In comparison to the positive control, treatment with calcium hydroxide (P = 0.000 and 0.000) or activ points (P = 0.000 and 0.002) produced significantly lower colony counts of E. faecalis at dentine depths of 100 and 250 µm, respectively. Calcium hydroxide (2.10 × 102 CFU mL,1) was significantly more effective than activ points (1.58 × 103 CFU mL,1) at 100 µm (P = 0.013), but not at 250 µm (P = 0.353). Neither of these two medications was able to eliminate E. faecalis completely. Conclusions, Chlorhexidine-impregnated activ points did not possess an in vitro inhibitory activity strong enough to eliminate E. faecalis completely from infected dentinal tubules. [source] Impact of multiple HPV infection on response to treatment and survival in patients receiving radical radiotherapy for cervical cancerINTERNATIONAL JOURNAL OF CANCER, Issue 3 2002Barbara Bachtiary Abstract To obtain information on the incidence and the clinical significance of infection with various types of the human papillomavirus (HPV) in cancer of the uterine cervix, we retrospectively examined the HPV status of 106 patients who had received radical radiotherapy for cervical cancer stages IB to IIIB. DNA was extracted from formalin-fixed, paraffin-embedded biopsies and PCR was carried out to identify HPV types 16, 18, 31, 35, 33 and 45. To detect additional HPV types, consensus PCR products were cloned and sequenced. A catalyzed signal-amplified colorimetric in situ hybridization was carried out in 84 of 106 specimens as a positive control. Response to therapy, progression-free survival (PFS) and cervical cancer-specific survival (CCSS) were the statistical endpoints. Survival analysis was carried out using univariate and multivariate analysis (Cox regression). Ninety-six patients (90.6%) were HPV-positive and 42/96 (43.7%) were positive for multiple HPV types. Eight patients had persistent disease after radiotherapy. From these 8 patients, 7 were infected with multiple HPV types and only 1 patient had an infection with a single HPV type. After a median follow up period of 50 months, patients with multiple HPV infection had a significantly shorter PFS and CCSS compared to those with single HPV infection (24.8% and 34.9% vs. 64% and 60.8%, Log rank, p < 0.01 and 0.04). In multivariate analysis, the presence of multiple HPV types (RR 1.9), node status (RR 2.3), tumor size (RR 3.2) and histologic type (RR 4.8) were independent prognostic factors of CCSS. Our results demonstrate that the presence of multiple HPV types is associated with poor response and with reduced survival in cervical cancer patients who receive radiotherapy as the primary treatment. © 2002 Wiley-Liss, Inc. [source] Cosmeceutical properties of polysaccharides from the root bark of Ulmus davidiana var. japonicaINTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 2 2007Sang Yong Eom In Korea and China, Ulmus davidiana var. japonica has been used as a traditional oriental medicine for the treatment of difficulty in urination, skin inflammation, etc. In order to investigate the potential of a polysaccharide extract from Ulmus davidiana var. japonica as a cosmetic ingredient, we measured its moisturizing effect, photo-induced cytotoxicity, and anti-inflammatory effect. After hydrolysis, HPLC experiments showed that the composition of the polysaccharide extract was mainly rhamnose, galactose, and glucose. The molecular weight of the obtained Ulmus davidiana root extract was 20 000. The intrinsic viscosity was 90 dL/g. In a moisturizing test conducted through the measurement of water loss in a desiccator and of moisture content with a Corneometer CM820, Ulmus davidiana root extract showed almost the same moisturizing effect as hyaluronic acid. In an assay for inhibition of the H2O2-activated release of PGE2, IL-6, and IL-8 in normal human fibroblast cell lines, Ulmus davidiana root extract showed an inhibitory activity of PGE2 release in a dose-dependent manner (up to 85.9% at a concentration of 0.1%). The percent inhibition of the release of IL-6 was in the range of 45.6,64.5% (H2O2 was used as the positive control). Moreover, the release of IL-8 was completely inhibited in the entire concentration range (>0.0025%). In a test of recovery from photo-induced damage after UVA irradiation (3 J/cm2), the cell recovery of human fibroblasts increased to levels two times higher than that of the positive control, which was UVA-damaged cells in the absence of Ulmus davidiana root extract (up to 60.2% at 3.0% of Ulmus davidiana root extract). In a photo-induced cytotoxicity assay in the presence of promethazine as a photosensitizer, Ulmus davidiana root extract showed approximately 48% of the increased cell viability of the control. Therefore, Ulmus davidiana root extract may be useful for the development of a cosmetic ingredient. [source] | |||||||||||||||||||||||||||||||||||||||||||