The OECD Validation Program of the H295R Steroidogenesis Assay for the Identification of In Vitro Inhibitors and Inducers of Testosterone and Estradiol Production. Phase 2: Inter-Laboratory Pre-Validation Studies (8 pp)

Background, Goals and Scope In response to concerns that have been raised about chemical substances that may alter the function of endocrine systems and result in adverse effects on human health, an OECD initiative was undertaken to develop and validate in vitro and in vivo assays to identify chemicals that may interfere with endocrine systems of vertebrates. Here we report on studies that were conducted to develop and standardize a cell-based screening assay using the H295R cell line to prioritize chemicals that may act on steroidogenic processes in humans and wildlife. These studies are currently ongoing as part of the ‘Special Activity on the Testing and Assessment of Endocrine Disruptors’ within the OECD Test Guidelines Program to review, develop, standardize, and validate a number of in vitro and in vivo toxicological assays for testing and assessment of chemicals concerning their potential to interact with the endocrine system of vertebrates. Study Design Six laboratories from five countries participated in the pre-validation studies. Each laboratory tested the effects of three model chemicals on the production of testosterone (T) and estradiol (E2) using the H295R Steroidogenesis Assay. Chemicals tested were well described inducers or inhibitors of steroidogenic pathways (forskolin, prochloraz and fadrozole). All experiments were conducted in 24 well plates following standard protocols. Six different doses per compound were analyzed in triplicate per plate. A quality control (QC) plate was run in conjunction with the chemical exposure plate to account for inter-assay variation. Each chemical exposure was conducted two or three times. Results All laboratories successfully detected increases and/or decreases in hormone production by H295R cells after exposure to the different model compounds and there was good agreement in the pattern of response for all groups. Forskolin increased both T and E2 while fadrozole and prochloraz decreased production of both hormones. All chemicals affected hormone production in a dose-dependent manner with the exception of fadrozole which caused maximum inhibition of E2 at the two least concentrations tested. Some inter-laboratory differences were noted in the alteration of hormone production measured in chemically exposed cells. However, with the exception of the production of T measured at one laboratory in cells exposed to forskolin, the EC₅₀s calculated were comparable (coefficients of variation 34–49%) for all hormones. Discussion and Perspectives The results indicated that the H295R Steroidogenesis Assay protocol was robust, transferable and reproducible among all laboratories. However, in several instances that were primarily related to one laboratory there were unexplained minor uncertainties related to the inter-laboratory hormone production variation. Based on the findings from this Phase 2 prevalidation study, the H295R Steroidogenesis Assay protocol is currently being refined. The next phase of the OECD validation program will test the refined protocol among the same group of laboratories using an extended set of chemicals (∼30) that will include positive and negative chemical controls as well as a broad spectrum of different potential inducers and inhibitors of steroidogenic pathways. Submission Editor: Dr. Carsten Brühl (bruehl@uni-landau.de)     

Energetic expenditure during vocalization in pups of the subterranean rodent<Emphasis Type="Italic"> Ctenomys talarum</Emphasis>

Theoretical signaling models predict that to be honest, begging vocalizations must be costly. To test this hypothesis, oxygen consumption was measured during resting and begging (i.e., vocalizing) activities in pups of the subterranean rodent Ctenomys talarum by means of open-flow respirometry. No statistical differences in individual oxygen consumption between resting and calling pups ranging in age from day 2 to day 20 were found. Given these data, begging calls of C. talarum could not be considered as honest advertisements of offspring need, contrary to what suggested by the behavioral observations of the mother and pups during the nestling period.     

Umbilical cord pseudocyst in trisomy 18

Prenatal diagnosis of cord defects by means of ultrasound examination is possible and highly accurate. Although this is a rare pathological finding, we report two cases in which umbilical cord pseudocysts were associated with trisomy 18. These observations underscore the need of umbilical blood sampling for establishing the karyotype in fetuses with such umbilical cord anomalies and the importance of careful examination of placentas and infants born with such defects.     

Europe's last Mesozoic bird

Birds known from more than isolated skeletal elements are rare in the fossil record, especially from the European Mesozoic. This paucity has hindered interpretations of avian evolution immediately prior to, and in the aftermath of, the Cretaceous-Tertiary (K-T) extinction event. We report on a specimen of a large ornithurine bird (closely related to Ichthyornis) from the uppermost Cretaceous (Maastricht Formation) of Belgium. This is the first record of a bird from these historic strata and the only phylogenetically informative ornithurine to be recovered from the Mesozoic of Europe. Because this new specimen was collected from 40 m below the K-T boundary (approximate age of 65.8 Ma), it is also the youngest non-neornithine (=non-modern) bird known from anywhere in the world.     

WATERSHED WEIGHTING OF EXPORT COEFFICIENTS TO MAP CRITICAL PHOSPHOROUS LOADING AREAS1

ABSTRACT: The Export Coefficient model (ECM) is capable of generating reasonable estimates of annual phosphorous loading simply from a watershed's land cover data and export coefficient values (ECVs). In its current form, the ECM assumes that ECVs are homogeneous within each land cover type, yet basic nutrient runoff and hydrological theory suggests that runoff rates have spatial patterns controlled by loading and filtering along the flow paths from the upslope contributing area and downslope dispersal area. Using a geographic information system (GIS) raster, or pixel, modeling format, these contributing area and dispersal area (CADA) controls were derived from the perspective of each individual watershed pixel to weight the otherwise homogeneous ECVs for phosphorous. Although the CADA‐ECM predicts export coefficient spatial variation for a single land use type, the lumped basin load is unaffected by weighting. After CADA weighting, a map of the new ECVs addressed the three fundamental criteria for targeting critical pollutant loading areas: (1) the presence of the pollutant, (2) the likelihood for runoff to carry the pollutant offsite, and (3) the likelihood that buffers will trap nutrients prior to their runoff into the receiving water body. These spatially distributed maps of the most important pollutant management areas were used within New York's West Branch Delaware River watershed to demonstrate how the CADA‐ECM could be applied in targeting phosphorous critical loading areas.     

Remote measurement of NO2 in the brown cloud over Albuquerque,New Mexico

Remote measurements of nitrogen dioxide (NO₂) were recorded in the brown cloud over Albuquerque, NM, using absorption spectroscopy in the winter of 1987-88 and summer of 1989. The NO₂ burdens (optical densities) measured in this manner were found to be in excess of 100 ppm-m. These long pathlength measurements correspond to total concentrations of approximately 5–10 ppb over the integrated observation pathlengths, which ranged from 10–20 km. These concentrations compare well with single location, independent NO _x analyses. Using two correlation (absorption) spectrometers simultaneously, it was shown that the NO₂ distribution is not uniform over the city and can change on the order of minutes in the boundary layer late in the day, demonstrating the advantages of NO₂ optical measurements for assessing the location and extent of urban nitrogen dioxide levels in the boundary layer.     

Host-plant green-leaf volatiles synergize the synthetic sex pheromones of the corn earworm and codling moth (Lepidoptera)

Summary The capture of adult male moths in female sex pheromone traps of two key agricultural pests, the corn earworm (Helicoverpa zea) and the codling moth (Cydia pomonella), is enhanced or synergized by a certain group of host-plant volatiles, the green-leaf volatiles (GLVs). Since female adults of both species call and release their sex pheromones while perched upon the leaves of their host-plants, the volatile constituents from the leaves of a number of host-plants were compared. Sex pheromone traps containing one of the prominent leaf volatiles of certainH. zea hosts, (Z)-3-hexenyl acetate, not only significantly increased the capture ofH. zea males but were preferred over traps baited only with sex pheromone. Similarly, traps baited with synthetic sex pheromome ofC. pomonella plus a blend of GLVs captured significantly more males than traps baited only with sex pheromone. Since male moths are not captured in traps baited only with these GLVs, it appears that these GLVs act as pheromone synergists which increase or enhance the attraction or arrestment of male moths in pheromone traps.     

Progress in teenage crash risk during the last decade

OBJECTIVE: The purpose of the present study was to examine the most recent data on teenagers' fatal and nonfatal crashes in the United States to determine current crash rates as well as changes in crash rates during the past decade METHODS: Data for calendar years 1996 and 2005 were extracted for fatal crashes from the Fatality Analysis Reporting System and for police-reported crashes from the National Automotive Sampling System/General Estimates System. To calculate crash rates, population data were obtained from the Census Bureau, and mileage data were obtained from the 2001 National Household Travel Survey RESULTS: During 2001-02, the latest year for which mileage data are available, 16 year-old drivers had higher fatal and nonfatal crash rates per mile traveled than all but the very oldest drivers. However, fewer 16 year-olds typically are licensed to drive and they drive fewer miles per year than all but the oldest drivers. Thus, their fatal and nonfatal crash rates per population in 2005 were lower than among other teenagers and among drivers 20-29. During the past decade the most progress has been made in reducing crashes among the youngest drivers. Between 1996 and 2005 both fatal and police-reported crashes per population declined about 40% for 16 year-old drivers, compared with about 25% for 17 year-old drivers and 15-19% for 18 year-old drivers. The greatest reductions for 16 year-olds occurred in nighttime crashes, alcohol-related fatal crashes, and fatal crashes involving multiple teenage passengers. CONCLUSIONS: Substantial progress has been made in reducing fatal and nonfatal crashes per population among 16 year-old drivers. Although this study was not designed to examine the role of graduated licensing, the results are consistent with the increased presence of such laws, many of which restrict nighttime driving and driving with teenage passengers. IMPACT ON INDUSTRY: Restrictions on nighttime driving and driving with teenage passengers should be made a part of all states' graduated licensing systems. Historically, 16 year-olds have had the highest crash risk per licensed driver and per mile traveled. Given the dramatic reductions in per population crash rates among 16 year-olds, it is possible that their per mile and per licensed driver rates also have declined and may no longer be as elevated relative to other ages. However, shortcomings in the licensed driver data and a lack of recent mileage data hamper our ability to examine these issues. If we are to continue to provide a yardstick against which we can measure progress among the youngest drivers, immediate steps need to be taken to restore the availability of reliable exposure data.     

Urban soil-lead (Pb) footprint: retrospective comparison of public and private properties in New Orleans

Lead (Pb) is a toxin that after childhood exposure poses a lifetime of health risks. One route of exposure is soil-Pb as a result of ∼12 million metric tons of Pb residue in paint and gasoline sold in the US during the 20th Century. Pb accumulated in soil of the community is a good predictor for blood Pb of children living there. This retrospective study compares the soil-Pb on Housing Authority of New Orleans (HANO) properties with adjacent private residential (RES) properties within a 0.8 km (0.5 mile) radius. The sample subset (n = 951) is from two soil-Pb surveys (total n = 9,493) conducted between 1989 and 2000. The properties were in both the inner city (CORE) and outlying (OUTER) communities. The data were analyzed using multi-response permutation procedures (MRPP). The soil-Pb results differ significantly (P-value < 0.001) on same-aged HANO properties at different locations; thus, year of construction does not give adequate explanation for the soil-Pb differences. HANO and RES soils are significantly more Pb contaminated in the CORE than in OUTER communities (P-value < 0.001). The CORE has many more years of traffic congestion than OUTER communities; therefore, the lead additives to gasoline, and not lead-based paint, best elucidate the differences of the soil-Pb footprint at HANO and RES properties in the CORE and OUTER communites. Currently HANO properties are being redeveloped with cleaner soil, but soil on RES properties in the CORE of New Orleans remains a large source of Pb (median = 707 mg/kg in this study) for human exposure, especially children.