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1] Singh J. Effect of protein, zinc, and carbon monoxide on daily food intake duringpregnancy and fetal weight in mice. Reprod Toxicol 2008.

oi:10.1016/j.reprotox.2010.05.076

otential teratogenicity of anticonvulsants Gabapentin and Val-roic acid in C-F rats

rishna P. Singh ∗, Kiran Gupta

Neurobiology Lab., Dept of Zoology, University of Allahabad, AllahabadU.P.), India

For treatment of epileptic cases, both first and second genera-ion AEDs (antiepileptic drugs) are available but physicians alwaysace difficulties to manipulate the drug doses during pregnancyonsidering the potential benefits to mother and possible risks toeveloping fetuses. Among the typical AEDs, most of the drugs areeratogenic in nature but teratogenic safety of the newer AEDsncluding GBP (Gabapentin) has not been well established so far.hough, some reports are available but are contradictory and incon-lusive and limited to gross teratological observations. Keepinghese views in consideration, present study has been planned tovaluate the teratogenic safety of GBP and compare it with VPAValproic acid). As per study design pregnant C-F rats of differ-nt groups were exposed to GBP (300 and 400 mg/kg/bw) and VPA50 and 100 mg/kg/bw) from GD0–20. Similarly, vehicle exposedams were maintained as control. 50% pregnant dams of each groupere sacrificed at GD21 and rest of the dams were allowed toeliver naturally and their offspring were reared up to PND 56.hese drugs induced fetal toxicity as external gross malformations,keletal anomalies of limbs and ribs, fetal body weight and braineight. Histopathological evaluation of AEDs exposed fetal brain

evealed significant disturbances of neuroarchitectural pattern ofifferent neuronal layers, reduced neuronal density of differentrain regions (Cerebral cortex, Hippocampus and Striatum). Prena-ally exposed offspring not only showed neurodevelopment delaynd reduced growth pattern from PND1–56 but also displayed long-asting effect of these drugs on neurobehavioural disturbances likenxiety, cognition in young-adult rat offspring. This study con-ludes that GBP is less teratogenic than VPA as gross birth defectsut GBP is equally teratogenic at neuroanatomical and neurobe-avioural level. It is hypothesized that mechanism of action for

nducing teratogenic responses may be through developmental dis-urbances of fetal neurotransmitters (NTs), hence role of NTs asrophic factor may not be ruled.

oi:10.1016/j.reprotox.2010.05.077

isruption of microtubule assembly: A common teratogenicechanism for two new structural alerts

ashir Surfraz a,∗, Martin P. Payne a, Robert D. Benz b,dwin J. Matthews b

Lhasa Limited, 22-23 Blenheim Terrace, Woodhouse Lane, Leeds LS2HD, United KingdomInformatics and Computational Safety Analysis Staff, Office of Phar-aceutical Science, Center for Drug Evaluation and Research, U.S.

ood and Drug Administration, 10903 New Hampshire Avenue, Silverpring, MD 20993-0002, United States

Several approaches exist for the in silico prediction of terato-enicity from chemical structure [1]. A knowledge-based approachas potential for transparency, with interpretable explanationsccompanying predictions. In this work we describe two of twelve

ogy 30 (2010) 233–248 245

chemical classes investigated which resulted in novel structuralalerts suitable for in silico prediction of teratogenicity, a potentialalternative to animal testing. Initial structural clustering analysis ofthe Food and Drug Administration ICSAS dysmorphogenesis datasetfor the mouse, rabbit and rat suggested an investigation of theteratogenicity of chemicals with an indole or indole-like moiety.A detailed review of public domain information was undertakenwhich included results of toxicity and mechanistic studies sup-plemented where appropriate with details of chemical, metabolicand pharmacological properties. Human expert analysis of thecollated data allowed development of two structural alerts, ben-zimidazole derivatives and vinca alkaloids. Both classes can beteratogenic in various animal models and share a common pat-tern of malformations including; craniocerebral and craniofacialdefects, limb abnormalities, ocular malformations and a varietyof other skeletal defects. Evidence to confirm that vinca alkaloidsor benzimidazole derivatives can cause birth defects in humans ispresently equivocal. The detailed mechanism by which these com-pounds can give rise to teratogenicity has not been fully established,but is considered to occur through a similar pathway consistentwith the commonality of observed malformations. Vinca alkaloidsare known to disrupt microtubule assembly [2] and benzimida-zole derivatives are potent inhibitors of tubulin polymerisation,interacting at the colchicine-binding domain [3]. Both sets of chem-icals ultimately prevent spindle formation and cause mitotic arrest.Unsurprisingly, colchicine itself is a known teratogen and a micro-tubule growth inhibitor. Benzimidazoles can also inhibit cartilageproteoglycan synthesis and therefore chondrogenesis, a potentialmechanism that can contribute to their teratogenicity. Structuralalerts for teratogenicity may be associated with more than onemechanism of action. Furthermore, the confidence in the pre-diction from either structural alert is enhanced by the fact thatboth chemical classes can cause a similar pattern of defects bya common mechanism of action. These structural alerts exem-plify a cost-effective way to identify potential toxicity in novelcompounds.

References

1] Cronin MTD, Worth AP. QSAR Comb Sci 2008;27:91–100.2] Jordan MA, Wilson L. Nat Rev Cancer 2004;4:243–65.3] Whittaker SG, Faustman EM. Toxicol Appl Pharmacol 1992;113:144–51.

doi:10.1016/j.reprotox.2010.05.078

Cranio-facial anomalies in the Slovak Republic

Elena Szabová a, Dagmar Zeljenková a, Richard Molokác b,∗,Jevgenij Kovriznych a, Eva Véghová c, Daniela Brasenová d,Eva Nescáková e, Agáta Molnárová f, Jozef Fedeles f

a Slovak Medical University, Bratislava, Slovak Republicb 1st Department of Gynaecology and Obstetrics, L. Pasteur UniversityHospital, Kosice, Slovak Republicc Centre of the Clinical Genetics, Bratislava, Slovak Republicd National Health Information Center, Bratislava, Slovak Republice Dept of Anthropology, Faculty of Natural Sciences, Commenius Uni-versity, Bratislava, Slovak Republicf Dept of Plastic Surgery, Bratislava, Slovak Republic

E-mail address: elena.szabova@szu.sk (R. Molokác).Introduction: A birth defect is an abnormality of the struc-

ture, function, or metabolism present at birth that can causephysical or mental disability, or is fatal. It is generally estimatedthat around 14% of babies are born with a single minor mal-formation. The precise cause of congenital malformations is notknown for as many as 50–60% of total births. Birth defect risks

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n human population exposed to pesticides, drugs, pharmaceuti-als, or other chemicals have been continually studied. It has beenuggested that nutrition can play an important role in the man-festation of oral clefts. For example maternal periconceptionalse of folic acid has been found to reduce the risk of neural tubeefects.

Aim and methodology: A project plan for a specific birth defectsegister was established to collect information on typical oral cleftases, with the aim to have the best and widest information onpidemiology of oral clefts, such as birth prevalence, time trends,ifferences among populations, and in the next future, analysisf risk factors. Our results represent data which were collectedccording to the common protocol of the WHO project and sum-arize data from the years 2001–2008.Results and discussion: The true incidence of congenital mal-

ormations is difficult to determine because of inconsistent andncomplete data gathering. Our scientific group became a memberf the International Clearinghouse for Birth Defects Surveillancend Research This organization endeavours to eliminate the gap ofncomplete information among world-wide databases. Participat-ng surveillance programmes carry out systematic data collectionnd use the data for public health action, including comparison ofrevalence rates, advocacy, policy development and conduction ofpidemiological studies that investigate causes and consequencesf birth defects. We collected data on congenital malformationsnd joined them to the common database with the aim to stimu-ate existing databases and sharing their data, creating a specific

orldwide database. Cleft lips or palates occur in somewhereetween one in 600–800, births with incidence variations in dif-erent racial groups. In our study we have found that about 6% ofll birth defects are craniofacial anomalies. Families with a his-ory of cranio-facial anomalies (parents, other children or closeelatives), are more likely to have a child with an oral cleft. How-ver, many families without oral cleft history also have childrenith craniofacial or different anomalies. This fact evoked scien-

ists to believe that one of the causes is the interaction betweenpecific genes with patterns of normal palate closure and lipevelopment and environmental factors. Hence it is very impor-ant to evaluate the link between parent’s health conditions andnvironmental toxins in order to minimize exposure to harmfulompounds.

oi:10.1016/j.reprotox.2010.05.079

isphenol A exposure at low dose level affects early placentaevelopment in CD1 mice: Preliminary data of an integrated inivo/ex vivo study

oberta Tassinari ∗, Sabrina Tait, Francesca Maranghi, Albertoantovani

Department Food and Veterinary Toxicology Unit, Department Vet-rinary Public Health and Food Safety, Istituto Superiore di Sanità,taly

-mail address: roberta.tassinari@iss.it (R. Tassinari).Introduction: Bisphenol A (BPA) is a widespread chemical used in

lastic polymers also intended as food contact materials. RecentlyFSA published an opinion on the hazard and risk assessment ofPA migrating from polycarbonate materials in food [1], assumingTDI of 0.05 mg/kg bw based on a NOAEL of 5 mg/kg bw. However,

ome data suggests that BPA may cause adverse effects in rodents

t doses below the “NOAEL” used to determine the TDI. The presenttudy considered placental development as a potentially suscepti-le target of BPA. This is highly relevant for human health, withegard to both pregnancy outcome and embryo/fetal nutrition androgramming.

ogy 30 (2010) 233–248

Methods: The BPA effects at cellular and molecular levels duringpre-implantation and early embryogenesis stages were investi-gated, following oral administration of pregnant CD-1 mice with0 (vehicle only = corn oil), 0.5 (10% of NOAEL) or 50 (dose level withexpected endocrine effects) mg/kg bw/die of BPA from gestationalday (GD) 1 to GD11. At GD12, one half of the embryos and placentaswere fixed in 10% buffered formalin and prepared for histologi-cal analyses, while remaining placentas were stored (−80 ◦C) formicroarray analysis.

Results: No increase of dysmorphogenesis, growth retardationor embryonic resorptions was observed. Histopathological exami-nation of placentas revealed a significant dose-related increase indegeneration/necrosis of giant cells [60% at 0.5 mg/kg and 100% at50 mg/kg vs 0% in control group; p-value 0.0035] and accumulationof glycogen and lipids in the junctional zone [50% at 0.5 mg/kg and100% at 50 mg/kg vs 0% in control group; p-value 0.0034]. Microar-ray analysis of placentas was performed using the Agilent platformfeaturing the whole mouse genome. 181 and 132 genes were signif-icantly modulated at 0.5 and 50 mg/kg dose levels respectively (foldchange ± 1.5, p-value for moderated t statistic < 0.01, false discov-ery rate < 5%). The expression profiles at the two doses were quitedissimilar, affecting different groups of genes. The functional anal-ysis revealed different enrichments in Gene Onthology terms, withthe lower dose mostly implicated in angiogenesis, blood vessel andvascular development and biosynthetic processes connected withglycerol derivatives. The higher dose on the other hand, seemedto affect genes mostly involved in apoptotic, cell motility and cellstructure/organization processes, as well as in xenobiotics detoxi-fication.

Conclusions: Our results suggest that the 0.5 mg/kg bw dose level(10% of the NOAEL used for the TDI), may elicits tissue and molecu-lar alterations in mouse placenta. The molecular effects also suggesta different ability of BPA to impact the developmental program-ming depending on the dose. The actual relevance for human healthof these preliminary data has to be carefully evaluated; neverthe-less such findings might lend support to a reconsideration of thepresent TDI for BPA.

Reference

1] EFSA J 2008;759:1–10.

doi:10.1016/j.reprotox.2010.05.080

Incorporation of biochemical endpoints to teratogenesisevaluation: Acetylcholinesterase activity in valporic acid treatedzebrafish embryos

Elisabet Teixidó ∗, Ester Piqué, Jesús Gómez-Catalán, Joan M. Llo-bet

GRET-CERETOX, Toxicology Unit, Public Health Department, Univer-sity of Barcelona, Barcelona, Spain

Introduction: Embryotoxicity evaluation is carried out basicallythrough morphological endpoints. The incorporation of biochemi-cal endpoints represents mainly two advantages: the elucidationof possible mechanisms of action and a higher sensitivity thanmorphological endpoints in order to detect alterations. Acetyl-cholinesterase (AChE) is an enzyme that is found in central andneuromuscular synapses of the cholinergic system and is respon-sible for the degradation of the neurotransmitter acetylcholine.

An important role in the neuronal and muscular development,unrelated to its catalytic function, has been postulated due to itsstructural similarity with cell adhesion proteins. In consequence,the evaluation of AChE activity could be a useful parameter forneurotoxicity and delayed growth evaluation. The aim of this study