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1434 URINARY BIOMARKER DETECTION OF MELAMINE- CYANURIC ACID-INDUCED KIDNEY DAMAGE IN RATS. Q. Zhang 1 , G. Gamboa da Costa 2 , R. Beger 2 , L. Schnackenberg 2 , J. Sun 2 , L. Pence 2 , S. Bhattacharyya 2 , C. Jacob 2 , R. P. Brown 1 and P. L. Goering 1 . 1 CDRH, U.S. FDA, Silver Spring, MD and 2 NCTR, U.S. FDA, Jefferson, AR. Exposure of rats to melamine and cyanuric acid in the feed can result in a dose-dependent increase in formation of melamine-cyanuric acid crystals in the kidney. The aim of this study was to determine if urinary biomarkers of acute kidney injury can be used to detect noninvasively renal damage associated with crystal formation in the kidneys of melamine-cyanuric acid-exposed rats. Male and female Fischer 344 rats were fed a melamine-cyanuric acid mixed diet at approximately 5, 10 and 15 mg/kg body weight/day for 28 days. Urine samples were collected at 1, 2, 3, 4, 14, and 28 days. A series of urinary protein biomarkers (Kim-1, NGAL, osteopontin, alpha-GST, GSTYb1, RPA-1 and clusterin) were measured by MesoScale Discovery multiplex assay system. Urinary metabolites were evaluated by NMR and LC/MS metabolomic assays. The results showed that RPA-1 (distal tubule and collecting duct injury biomarker) was elevated at the 10 and 15 mg/kg doses in a timedependent manner; however, there were no changes observed in other urinary protein biomarkers at any other dose or time point. Metabolomic analysis showed increased levels of urinary proline on days 3 and 28 in male rats and day 28 in female rats at the 15 mg/kg dose only. An increase of urinary hydroxyproline was observed on day 28 in male rats and on day 1 in female rats at 15 mg/kg. Significant renal histopathological changes were observed at the 15 mg/kg dose, although variable levels of melamine-cyanurate crystal formation were observable at the lower dose levels. These data indicate that increased levels of urinary RPA-1 and various urinary amino acids are seen at doses of melamine-cyanuric acid that produce histopathological damage in the kidney. As a result, these endpoints may serve as promising noninvasive urinary biomarkers for the detection of acute kidney injury associated with melamine-cyanuric acid exposure. 1435 VALIDATION OF BEST DETECTION METHODS FOR MEASURING OXIDATIVE STRESS. M. B. Kadiiska. NIEHS/NIH, Research Triangle Park, NC. Many techniques now exist that potentially allow the measurement of oxidative stress in experimental animal models and humans. The concept of measuring oxidative stress status in humans has not been approached as a real possibility in clinical practice because standardized methods are not yet established. NIEHS/NIH has taken the lead in organizing and sponsoring the first national and international comprehensive comparative study for determining which of the available biomarkers of oxidative stress are most specific, sensitive and selective. Investigators from 25 laboratories are participating in this study, called BOSS. The goal of the BOSS is to validate and find the best markers of oxidative stress that are applicable to different oxidative insults and measurable in stored specimens. Antioxidants and various oxidative products of lipids, proteins and DNA were measured in blood, plasma and urine of experimental animals with three different oxidative insults of CCl4 poisoning, ozone exposure, and LPS treatment. It was found that plasma concentrations of MDA and F2-isoprostanes (measured by GC/MS) and urinary concentrations of isoprostanes (measured with ELISA) were increased in CCl4- and ozone-exposed rats, and LPS-treated mini pigs in a time- and dose-dependent manner. Antioxidants (GSH/GSSG, tocopherols, ascorbic acid, uric acid, and total antioxidant capacity), oxidation products of proteins (protein carbonyls, methioninesulfoxidation, and tyrosine oxidative products) and DNA (strand breaks, 8-OH-dG, M1G) were not significantly changed. It is concluded that measurements of plasma lipid peroxidation products MDA and isoprostanes (by GC/MS) and urinary isoprostanes (by ELISA or GC/MS) are promising candidates for general biomarkers of oxidative stress. Various antioxidants, protein- and DNA-oxidation products are not identified as biomarkers of oxidative stress induced by CCl4- , ozone- or LPS- exposures. 1436 STEROID HORMONE PROFILING BY MASS SPECTROMETRY IN RAT BLOOD COLLECTED VIA THE CULEX® AUTOMATED IN VIVO SAMPLING SYSTEM. C. Penno 1, 2 , L. Morawiec 1 , M. Schwald 1 , N. Zamurovic 1 , S. Busch 1 , A. Wolf 1 , F. Pognan 1 , S. Chibout 1 , A. Odermatt 2 and M. Dong 1 . 1 Preclinical Safety, Novartis, Basel, Switzerland and 2 Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland. Impaired steroid hormone regulation has been implicated in the pathogenesis of multitudinous drug-induced toxicities. A quantitative profile of steroid hormone metabolites holds great potential to help assessing drug safety, dissecting toxic mechanisms, understanding disease progress and providing rational therapeutic in- 308 SOT 2011 ANNUAL MEETING tervention. A novel LC-MS/MS method has been established to quantify a panel of 12 steroid hormones: aldosterone, cortisol, cortisone, corticosterone, 11-dehydrocorticosterone, 11-deoxycorticosterone, 11-deoxycortisol, testosterone, dihydrotestosterone, androstenedione, 17α-hydroxyprogesterone and progesterone (comprising rodent and human metabolites). Steroid metabolites were resolved by liquid chromatography using an Allure Biphenyl (Restek) column and quantified by Agilent 6410 Triple-Q mass spectrometer. The method presented excellent dilution linearity for all 12 metabolites and the limits of quantitation were ranging from 250 pM for androstenedione to 4 nM for corticosterone. Method validation and measurements were performed in 50 μL of rat plasma spiked with calibrators and deuterized internal standards. The method was then applied to profile timecourse changes of steroid hormones in rat plasma. Wistar male rats were connected to the Culex® system through carotid artery or jugular vein. Blood collection was programmed every 3 hours during a 24-hour period. Interim results indicated that changes of corticosterone, 11-dehydrocorticosterone, and 11-deoxycortisol followed the known circadian rhythm, suggesting the non-disturbance of glucocorticoid profiles in catheterized rats. Levels of testosterone, androstenedione and progesterone were rather constant regardless of the day and night shift. The other steroid metabolites analyzed were below the detection limit. Samples from additional animals are currently being analyzed to gain a high level of statistical power. 1437 SERUM CARDIAC TROPONIN I CONCENTRATION AS AN EARLY BIOMARKER OF MYOCARDIAL HYPERTROPHY IN THE ROSIGLITAZONE OR 3, 3’, 5- TRIIODO-L-THYRONINE TREATED RAT. M. Dunn 1 , D. Coluccio 1 , R. Fernandes 1 , D. Knapp 1 , R. Nicklaus 1 , G. Hirkaler 1 , W. Geng 1 , M. Sanders 1 , T. Singer 2 and I. Mikaelian 1 . 1 Nonclinical Safety, Roche, Nutley, NJ and 2 Nonclinical Safety, Roche, Basel, Switzerland. Sponsor: K. Kolaja. Background: The peroxisome proliferator-activated receptor gamma (PPARγ) agonist rosiglitazone has been linked to cardiac hypertrophy in rats and an increased risk of myocardial infarction in man. Exclusive to the heart, cardiac troponin (cTn) I is released in response to myocardial damage. Our goal was to investigate the link between cardiac hypertrophy and low level changes of serum cTnI concentrations following rosiglitazone or 3,3’,5-triiodo-L-thyronine (T3) treatment in the rat. Methods and Results: Serial blood samples from male rats were evaluated following dosing of rosiglitazone (80 mg/kg/day) or T3 (20 μg/kg/day) for 14 or 28 days, with 14 or 27 days of recovery. Histopathology and clinical chemistry, including measurement of cTnI concentrations with the Singulex Erenna® Ultrasensitive Immunoassay were performed. Compared to aged-matched vehicle treated controls, a treatment-duration dependent increase of heart weight to brain weight was observed in both rosiglitazone (10% and 17%) and T3 (21% and 30%) treated groups after 14 and 28 days of treatment, respectively, and this cardiac hypertrophy was found to be reversed after the recovery period. Across treatment groups and at all time points, a positive correlation was established between cTnI concentration and the degree of cardiac hypertrophy. Significant elevations of cTnI were quantified after 14 and 28 days of rosiglitazone treatment, in the absence of histomorphologic findings. As expected, T3 treatment induced marked and progressive elevations in cTnI concurrent with histiocytic infiltration of the myocardium after 14 and 28 days of dosing, and necrosis after 28 days of dosing. Conclusion: Our results demonstrate the sensitivity of low level cTnI concentration changes as indicative of myocardial hypertrophy which may occur in the absence of histomorphologic findings. These results lend support for serum cTnI concentrations as an early biomarker of cardiovascular liability. 1438 CHARACTERISATION OF THE PRODROMAL AND REVERSIBILITY VALUE OF NOVEL RENAL SAFETY BIOMARKERS IN GENTAMICIN-TREATED RATS. J. Wood 1 , A. Shoieb 1 , A. McGuinty 1 , P. Cleall 1 , D. Hardy 1 , D. Collins 1 , C. Dubray 1 , R. Williams 1 , A. Moody 1 , K. Lynch 2 , D. Ennulat 2 , T. Sellars 2 , T. Lambert 2 , S. Beushausen 3 , E. Harpur 4 , M. Guffroy 1 and D. Brees 1 . 1 Pfizer, Sandwich, United Kingdom, 2 GlaxoSmithKline, King of Prussia, PA, 3 Pfizer, St. Louis, MO and 4 Sanofi-Aventis, Alnwick, United Kingdom. Sponsor: D. Robinson Gravatt. The objectives of this ILSI-HESI sponsored study were to further define the value of the novel renal biomarkers during the prodromal and reversibility phases of renal injury, using Gentamicin as a test toxicant. Gentamicin was administered by subcutaneous injection once daily to groups of 10 male Sprague-Dawley rats at 50 mg/kg/day for 1, 4 or 10 days. Overnight urine samples were collected pre-dose (days -7 and -3), on Days 2, 5, 11 (dosing period), and on Days 18, 25, 32 and 39 (recovery period) and analysed for Kim-1, NGAL, Osteopontin, Albumin, Regucalcin, Beta-2-Microglobulin, Cystatin C, Clusterin, GST-α, GST-Yb1, RPA- 1, NAG, Creatinine and Protein. Microscopically, tubular epithelial degeneration/necrosis of the proximal convoluted tubules was observed on Day 5, peaked on
Day 11 and started to recover on Day 18. These changes correlated with statistically significant increases (p
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The Toxicologist Supplement to Toxi
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The Toxicologist Supplement to Toxi
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1 BIODEGRADABLE MATERIALS FOR TISSU
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that genetic toxicology data are ge
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well-orchestrated lung inflammation
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scribed in the literature. We have
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years ago). We will illustrate how
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involved in the biodegradation proc
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stem cells but rather a treatment i
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additional compound showed agreemen
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82 COMPARISON OF 3H-THYMIDINE INCOR
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irritants. While in practice these
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alleles are especially vulnerable t
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109 CD4+ T CELL INTRINSIC TNF RECEP
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118 TO STUDY THE SIGNAL TRANSDUCTIO
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PAHs, such as dioxins, are prevalen
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containing substituents and/or hydr
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146 COMPUTATIONAL MODELING FOR QT P
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suggest that the combination of too
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164 TRANSLOCATOR PROTEIN (18 KDA) (
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function as a metal binding protein
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laboratory has demonstrated that NR
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known. The aim of this study was to
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from 5 to 28 days in duration) in e
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positive cells, caspase-3 activatio
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creased level in the 46 kDa preproe
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invasiveness at concentrations repr
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thracene (DMBA,50 μg in acetone, a
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247 CO-CARCINOGENESIS OF N, N- DIET
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sponds to the endosomal dsRNA that
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ODD in both WT (maximum 177% of con
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Lastly, using p53siRNA cells, p53 w
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its receptor Mpl to exert its funct
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294 LOW LEVEL OF LEAD CAN INDUCE PH
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lunar surface presented potential h
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lar about cellular export mechanism
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DNA in the kidney medulla, grandula
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5.00 and 7.50 mg/kg/day by oral gav
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events and carcinogenesis. Here we
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tinization of cells of the hair fol
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358 CHARACTERIZATION OF GENOME-WIDE
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RNAi were also performed to identif
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376 A FUNCTIONAL CROSSTALK BETWEEN
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UGT1A gene induction, while this re
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tivity, specifically peroxisome pro
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and cytotoxic effects; however, its
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histone deacetylase that is necessa
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ment. Paradoxically, buthionine sul
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drug leflunomide and its major (A77
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442 GENE EXPRESSION AND MORPHOLOGIC
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451 INHIBITION OF THE MITOCHONDRIAL
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460 SULFORAPHANE PREVENTS ACETAMINO
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drophilic/lipophilic balance. Other
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pharmacokinetic and toxicological p
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489 USE OF ‘OMICS DATA TO IMPROVE
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498 APPLICATION OF AGE-SPECIFIC ADJ
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507 A DOSE VOLUME TOLERABILITY STUD
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involved the use of an acute inflam
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sorption in the gastrointestinal (G
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(ABC) transporters actively transpo
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545 BEHAVIORAL EFFECTS OF REPIN IN
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a blood pressure phenotype that res
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and inhalation exposure system that
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and lethality associated with these
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position, on vascular reactivity an
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therefore more accurate and reprodu
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commercial chrysotile product that
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elative to their controls. ST-depre
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ats. The majority of the studies fo
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in the China Jintan Child Cohort St
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corneum thickness, cellular epiderm
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645 EVALUATION OF THE IN VITRO EPIS
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654 TCDD ADSORBED ONTO NATURAL AND
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tion revealed no test article-relat
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671 INFLAMMATION AND TISSUE DAMAGE
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model, ethanol was found to inhibit
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689 ENHANCED SUPPRESSIVE FUNCTION O
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NAC + LPS yielded different levels
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707 LIFE-STAGE PBPK MODELS FOR MULT
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downstream of the apical endpoint o
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726 UPDATED ALUMINUM PHARMACOKINETI
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global parameter sensitivity analys
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and renal inflammation induced by 2
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754 PLASMA, URINE, AND TISSUE CONCE
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cently characterized transporter OA
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exposure system was developed from
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lood cell mass and decrease in urin
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practical alternatives to MC in non
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imals dosed with 167 mg/kg THU alon
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and organ weights, clinical signs a
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yet is induced in most bladder and
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830 RISK ASSESSMENT OF THE SPILL: C
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knowledgebase creation. Results are
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852 UNDERSTANDING STRUCTURAL AND PH
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for integrating epidemiology and an
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motor activity, including age of th
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y partial purification of urine (fr
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pacity for self-renewal and may dif
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sue. The depth of our analysis led
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910 IMPLEMENTING CALIFORNIA’S GRE
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concentrations in six tissues and b
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934 THE FDA’S LIVER TOXICITY KNOW
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943 GENOME-WIDE DNA METHYLATION DIF
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membrane localization and subsequen
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trols, respectively. Subtoxic and t
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survival using both smoking regimes
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as non-, weak, moderate, or strong
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988 NNK-INDUCED CYTOKINE ALTERATION
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group (one-way ANOVA, p90 % viabili
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ered as an organ involved in xenobi
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Transport of CPZ across the Caco-2
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estrous cycle and sexual behavior d
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mRNA expression levels. Collectivel
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1043 THE EFFECTS OF ENDOCRINE DISRU
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1052 MONO-2-ETHYLHEXYL PHTHALATE IN
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Results: MC was variable within and
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UtSMCs. Phosphorylation of FoxO1 wa
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nonpregnant and pregnant diabetic m
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1089 PFOA-INDUCED LIVER EFFECTS IN
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events in the liver. AZD9056 was ev
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The peppermint oil caused a concent
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OA did not affect food consumption.
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1126 PERSISTENT DEVELOPMENTAL ARYLH
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1135 BACH2 BINDING TO Prdm1 AS A ME
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The purpose of this project was to
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one marrow. Since Chk1 is an attrac
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1163 THE MRNA AND MIRNA PROFILE OF
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have now compared the IC50 values b
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1181 ELUCIDATION OF FACTORS DETERMI
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enced by pre-exposure to cigarette
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Page 293 and 294: mice, each with 4 dose groups. One
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Page 297 and 298: third tetratricopeptide repeats (TP
Page 299 and 300: 7d. 28d after TMT injury there was
Page 301 and 302: subunits. Each cell type was treate
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Page 345 and 346: gene expression post-transcriptiona
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Page 355 and 356: were collected from TK animals at d
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showed similar levels of apoptosis
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1676 TWO BIOMARKERS FOR EVALUATION
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motifs selected. This system was ap
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1693 VOC SERUM LEVELS IN THE GENERA
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1702 DOES THE CLOCK MAKE THE POISON
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those with high nickel content are
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isk assessment. However, unique cha
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model of APAP metabolism and glutat
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logically & morphologically similar
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posure, blood chemistry was analyze
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elated to oxidative stress by measu
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ostasis), but work is needed to tra
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tokine products during carcinogenes
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ways as chemical stressors and, the
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toxicity of nanomaterials relates s
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1815 MEASURING COMPOUND SELECTIVITY
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1825 EFFECTS OF METABOLISM BY INTES
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cyanoacetic acid increased 2-3 fold
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1845 IS THE PROMISCUITY OF THE ARYL
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crorarray analysis. RT-PCR results
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are a family of phase-II biotransfo
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ous labs. As a result of positive s
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down the doses may produce more tox
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spectively. Below 100 mg/l of gemfi
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1900 GENERATION OF PRECISION CUT LI
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iomarkers or observation of lesions
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creased reticulocytes and lymphocyt
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statistically significant interacti
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monize sample preparation and analy
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NPC and sinonasal cancer in neighbo
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showed incidences of lung and nasal
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utylbenzenes, exhibited most simila
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1975 DIFFERENTIAL MODULATION OF CYP
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organic As to MMA(V) and a lower ca
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ole in invasion and metastasis of c
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3T3-L1 cells to low-levels of arsen
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2011 IDENTIFICATION OF A NOVEL VX M
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This work was supported by Cooperat
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2029 EFFICACY OF ENDOTRACHEAL ADMIN
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Diazepam injections and its combina
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2047 ESTERASE ACTIVITIES AND HEMOST
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nanoparticle-induced toxicity progr
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house, were iv infused into rats ov
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een explored. This study investigat
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croscopic analysis revealed that on
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egg yolk collected from turtles inh
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consistency of results in both expe
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2111 CYTOCHROME P450 3A5 GENOTYPE I
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esidues of organophosphates or thei
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manganism. Recent work from our lab
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Aβ1-40 in CSF and hippocampus in P
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2147 CATALASE MANIPULATIONS MODIFY
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ation based on real-world exposure
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NPs. Aggregation of citrate-stabili
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2175 THE ROLE OF SURFACE CHEMISTRY
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seen rapid uptake in biological ima
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effect on uterine weight or vaginal
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dicating widespread exposure. While
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components into the liver parenchym
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2223 ISOLATION AND DETECTION OF RIC
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stored (-20 celsius degree). The co
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2241 DDA, A BIOMARKER FOR ENVIRONME
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2249 INTERACTION BETWEEN DIETARY SE
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2258 PHARMACOKINETICS, EXCRETION BA
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2267 SENSITIVE AND SPECIFIC FLUORES
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2276 METABOLISM AND DISPOSITION OF
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ment of hypertension in companion a
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for the propyl series can be used f
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2304 IN VITRO EXPOSURE AND EVALUATI
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2313 THE SYNERGISTIC EFFECT OF SODI
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genes that play a crucial role in M
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2330 THE ROLE OF TRANSFORMING GROWT
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ined following up to 96 h continuou
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effect doses obtained with the rat
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diated transcriptional response of
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docrine disruptor activities of EDC
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individuals. Week 6 results were qu
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functionality of photosystem II and
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wild mice (Mus musculus) from areas
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2406 TOXICITY AND BIOACCUMULATION O
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Isocitric acid is the acid in the h
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2425 EFFECTS OF FUMONISINS IN ETHAN
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centration(μg/g) were: 5.1-95.3; 2
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2445 AUTOPHAGY IN DEVELOPMENT: A LI
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sures to inhaled Mn in dust. Nevert
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2466 CRITICAL EVALUATION OF ANIMAL
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elationships predict that resting r
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2489 COMPARATIVE TOXICITY OF BIODIE
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2497 TRANSCRIPTIONAL REGULATION OF
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2506 TOXICOLOGICAL CONSIDERATIONS O
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launched with the aim of developing
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forms mRNA expression levels were a
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2534 CUTANEOUS WOUND HEALING IN THE
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wells (0, 1, 5, 10, 25, 50, 100, an
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2553 AN ORGANOTYPIC MICROLIVER PLAT
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serum-free media. At 24 hrs, the gr
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2572 INCREASED ARSENIC BIOACCESSIBI
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nology to develop improved methods.
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tuna, swordfish, or mako shark (3.5
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nificantly reduce circulating thyro
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grouped into 9 observation periods
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histopathological or biochemical ev
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exhibited a hyperactive phenotype,
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the search of effective drugs for e
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2644 COVALENT BINDING OF 14 C 2-MET
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aries targeting all transcription f
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(p 9 weeks) and CSC phenotype acqui
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2673 IMMUNE-MEDIATED VASCULAR INJUR
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for risk identification and charact
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to control toxicant delivery in tob
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Author Index (Continued) The numera
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Keyword Index (Continued) Arsenite
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Keyword Index (Continued) Covalent
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Keyword Index (Continued) flow cyto
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Keyword Index (Continued) innate im
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Keyword Index (Continued) nanoparti
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Keyword Index (Continued) preservat
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Keyword Index (Continued) Thrombocy
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Toxicological Sciences The Official
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The Toxicologist - Society of Toxicology

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