Determining gestational age using genome methylation profile: A novel approach for fetal medicine

Gestational age determination by traditional tools (last menstrual period, ultrasonography measurements and Ballard Maturational Assessment in newborns) has major limitations and therefore there is a need to find different approaches. In this study, we looked for a molecular marker that can be used to determine the accurate gestational age of the newborn. To this end, we performed reduced representation bisulfite sequencing (RRBS) on 41 cord blood and matching placenta samples from women between 25 and 40 weeks of gestation and generated an epigenetic clock based on the methylation level at different loci in the genome. We identified a set of 332 differentially methylated regions (DMRs) that undergo demethylation in late gestational age in cord blood cells and can predict the gestational age (r = −.7, P = 2E-05). Once the set of 411 DMRs that undergo de novo methylation in late gestational age was used in combination with the first set, it generated a more accurate clock (R = .77, P = 1.87E-05). We have compared gestational age determined by Ballard score assessment with our epigenetic clock and found high concordance. Taken together, this study demonstrates that DNA methylation can accurately predict gestational age and thus may serve as a good clinical predictor.     

An experimental study of the effect of diet on the fatty acid profiles of the European Cuttlefish (<Emphasis Type="Italic">Sepia officinalis</Emphasis>)

Fatty acid analysis is an alternative dietary investigation tool that complements the more traditional techniques of stomach content and faecal analysis that are often subject to a wide range of biases. In applying fatty acid analysis to ecosystem studies, it is important to have an understanding of the effect diet has on the fatty acid profile of the predator. A feeding experiment, using crustacean and fish as prey for the European cuttlefish Sepia officinalis, was conducted to evaluate the effect of prey fatty acids on the fatty acid profile of this marine predator. Cuttlefish were fed on a fish diet for the first 29 days, and then changed to a crustacean diet for a further 28 days. Another group of cuttlefish was fed on a crustacean diet for the first 29 days, and then changed to a fish diet for a further 28 days. An analysis of the cuttlefish digestive gland showed that the fatty acid profile reflected that of the prey, with cuttlefish on a crustacean diet being clearly distinguishable from the cuttlefish on a fish diet. Cuttlefish fed on a fish diet for 29 days prior to the switch in diet were comparatively higher in 16:0, AA, 20:1ω9, DPA6, DHA, 22:4ω6 and DPA3 than those fed on crustaceans. Cuttlefish fed on a crustacean diet for 29 days prior to the switch in diet were comparatively higher in 17:1ω8, 18:1ω9, 18:2ω6, 18:1ω7, EPA and 20:2ω6 than those fed on fish. Following a change in diet, the fatty acid profile of the cuttlefish digestive gland reflected that of the new diet within 14 days. The results confirm that the fatty acid profile of the cuttlefish digestive gland clearly reflects the profile of its recent diet. It also shows that the digestive gland may not be an organ that accumulates dietary lipids for long-term storage, but rather is an organ where lipids are rapidly being turned over and potentially excreted.     

Rapid onset of mate quality assessment via chemical signals in a woodland salamander (<Emphasis Type="Italic">Plethodon cinereus</Emphasis>)

In the competition to acquire mates, environmental factors can be important in determining the relative quality of an individual. These aspects of quality are often conveyed through signals used for mate assessment by the most energetically invested sex. In red-backed salamanders, Plethodon cinereus, chemical signals communicate a surprising amount of information, much of which is dependent on the current condition of the sender. One such condition-dependent aspect of information conveyed via chemicals is the quality of the sender’s diet, but it is unclear as to whether this information is actively advertised by the sender (i.e., a signal) or simply inferred from food-derived odors (i.e., a cue). The amount of time on different diets required for changes in signaling is also unknown. I examined how quickly gravid female salamanders could detect a difference between the scents of males on high- vs. low-quality diets without fecal cues. The amount of protein present in two known signaling glands (the mental and postcloacal glands) was also measured after experimental feeding. Gravid females were able to infer the differences in male diet quality after only 1 week. Females also responded to the male scents more quickly after differential feeding had begun. High-quality males had significantly more protein present in both signaling glands than low-quality males. This scenario highlights the ongoing interplay between the quality of an individual and its environment, with males actually advertising the status of this relationship as an honest signal for mate assessment.     

Entwicklung und Einsatz eines standardisierten Labortests für Dungkäfer (Aphodius constans) zur Prüfung der Ökotoxizität von Veterinärpharmaka

Goal, Scope, and Background

According to European legislation, an environmental risk assessment of veterinary pharmaceuticals for dung beetles is required if the substance is a parasiticide for the treatment of pasture animals. However, the demonstration of the environmental safety of those substances for dung beetles is strongly hampered by the fact that no standardized test system is available so far. Therefore, starting with recommendations from the SETAC advisory group DOTTS (Dung Organism Toxicity Test Standardization), a test system using the widespread temperate dung beetle species Aphodius constans was developed.

Methods

Using experiences gained at the University of Montpellier (e.g. in field studies) and in other terrestrial tests, details of culturing and testing of A. constans were investigated in a project sponsored by the German Environment Protection Agency (UBA). Basically, the survival of young larvae of A. constans exposed to the test substance spiked into formulated (i.e. dried, grounded and re-wetted) or fresh dung is measured over a period of three weeks. Using the model substance dimethoate (an insecticide required as positive control in tests with the predatory mite Hypoaspis aculeifer) the suitability of the new test was checked. Afterwards, the new test method was used to determine the toxicity of four common veterinary pharmaceuticals (ivermectin, dicyclanil, moxidectin, praziquantel) in formulated and fresh dung. Finally, the effects of ivermectin on the dung beetles were determined in a test using fresh dung from treated cattle.

Results and Conclusions

For Dmethoate LC₅₀-values were found in a range of 1–3 mg a.s./kg dung (dew.), depending on the dung type. Praziquantel did not show eirecis in the tested concentration range (i.e. up to 1000 mg a.s./kg Dung (DW). The LC₅₀-values of the other three pharmaceuticals were very low (<6 mg a.s./kg dung (DW)). With the exception of Dicyclanil no differences between formulated and fresh dung were identified. In the case of ivermectin the LC₅₀-value were even lower: about 0.5 mg a.s./kg dung (DW). No differences between the results of test in which the substance was spiked into dung and those tests which were performed with dung from treated cattle were found. These results prove the sensitivity as well as the robustness of this test method.

Recommendations and Perspectives

Based on the experiences reported here, it is recommended to standardize this test in an international ring test so that it can be incorporated in the risk assessment process for veterinary pharmaceuticals as described in the currently developed international registration guidelines.     

The effect of perceived female parasite load on post-copulatory male choice in a sex-role-reversed pipefish

The last several decades of research in behavioral ecology have resulted in a deeper appreciation of post-mating processes and sexual conflict in sexual selection. One of the most controversial aspects of sexual selection is cryptic mate choice. Here, we take advantage of male pregnancy in a sex-role-reversed pipefish (Syngnathus typhle) to quantify cryptic choice based on perceived parasite load and other sources of variance in female fitness. Studies have shown that S. typhle males preferentially mate with females with lower parasite loads and that a male’s perception of female parasite load can be altered by tattooing females. We manipulated the apparent parasite load of females in controlled mating experiments to test the hypothesis that post-copulatory sexual selection is dependent on a male’s perception of female parasite load in pipefish. Our results provided no evidence for cryptic male choice based on perceived female parasite load. However, we found evidence that eggs from larger females were more likely to result in viable offspring than eggs from smaller females and that the first female to mate with a male transferred more eggs per copulation on average. Overall, our results show that potential for post-copulatory sexual selection does exist in pipefish, but the male’s perception of female parasite load does not play a major role in this process.     

Modelling above-ground herbage mass for a wide range of grassland community types

Whereas it is recognized that management of plant diversity can be the key to reconciling production and environmental aims, most grassland models are tailored for high-value grass species. We proposed to adapt a mono-specific grass model to take into account specific features of species-rich permanent grasslands, especially over the reproductive phase. To this end, we used the concept of plant functional type (PFT), i.e. the grouping of plant species according to plant traits determined by the response of plant species to different management practices (land use and fertilization) and characterizing of agronomic properties of the corresponding species. In the model, weather and nutrient availability act upon rates of biophysical processes (radiation capture and use, plant senescence). These rates are modified over times due to PFT-specific parameters determined experimentally which represent the different strategies of plant species regarding growth. The integration of these parameters into the model made it possible to predict herbage biomass accumulation rate under different management practices for a wide range of plant communities differing in their PFT composition. The model was evaluated in two steps, first by analyzing separately the effects of PFT and an indicator of nutrient availability on herbage accumulation and then by conducting a sensitivity analysis. It was validated using two independent datasets; a cutting experiment running over the whole growing season to examine the consistency of the model outputs under different cutting regimes, and a monitoring of meadows and pastures in spring over a whole growth cycle to assess the model’s ability to reproduce growth curves. Although a good fit was observed between the simulated and observed data, the few discrepancies noticed between field data and predicted values were attributed mainly to the potential presence of non-grass species. More specifically, we noticed that nutrient (mainly nitrogen) availability is the main driver of plant growth rate, and that PFT determines the times at which this rate changes in relation to the phenological characteristics of species present. We concluded that integration of the PFT concept into the initial mono-specific growth model is especially suited to evaluating the consequences of management practices on species-rich permanent grasslands to meet feed production targets.     

Origin of 2-ethylhexanol as a VOC

2-Ethylhexanol has been identified as a volatile organic compound (VOC) that contributes to the deterioration of indoor air quality. Plasticizers are common components of dust and building materials and are shown to be degraded by a variety of bacteria and fungi to produce 2-ethylhexanol and other metabolites. Of these, the 2-ethylhexanol has significant volatility and was observed in appreciable quantities. The degree to which 2-ethylhexanol is observed as a VOC in air samples would be limited by the fact that many of the microorganisms that are capable of producing this compound are also able to oxidize it to 2-ethylhexanoic acid, which is much less volatile. It is argued that an abiotic degradation mechanism of plasticizers that results in the generation of 2-ethylhexanol is unlikely and, if this did occur, other metabolites should have been observed. Thus, the microbial degradation of plasticizers is the most likely source of 2-ethylhexanol in indoor air.     

Extraction properties of new polymeric sorbents in SPE/GC analysis of phenol and hydroquinone from water samples

The influence of the polymers' properties on the solid-phase extraction recovery was studied. Different porous copolymers were used: di(methacryloyloxymethyl)naphtalene-divinylbenzene with ester functional groups (DMN-DVB), 4,4'-bis(maleimido)diphenylmethane-divinylbenzene with imide functional groups (BM-DVB), p,p'-dihydroxydiphenylmethane diglycidyl methacrylic ester-divinylbenzene (MEMDE-DVB) and p,p'-dihydroxydiphenylpropane diglycidyl methacrylic ester-divinylbenzene (MEDDE-DVB) with ester end hydroxyl functional groups. The extraction properties of new synthesized polymeric sorbents were compared with these of two commercial polymeric sorbents: styrene-divinylbenzene (SDB-1) and styrene-divinylbenzene with modified surface (StrataX). These sorbents were used in the SPE of phenol and hydroquinone from water samples.     

A flexible spatio-temporal model for air pollution with spatial and spatio-temporal covariates

The development of models that provide accurate spatio-temporal predictions of ambient air pollution at small spatial scales is of great importance for the assessment of potential health effects of air pollution. Here we present a spatio-temporal framework that predicts ambient air pollution by combining data from several different monitoring networks and deterministic air pollution model(s) with geographic information system covariates. The model presented in this paper has been implemented in an R package, SpatioTemporal, available on CRAN. The model is used by the EPA funded Multi-Ethnic Study of Atherosclerosis and Air Pollution (MESA Air) to produce estimates of ambient air pollution; MESA Air uses the estimates to investigate the relationship between chronic exposure to air pollution and cardiovascular disease. In this paper we use the model to predict long-term average concentrations of \(\text {NO}_{x}\) in the Los Angeles area during a 10 year period. Predictions are based on measurements from the EPA Air Quality System, MESA Air specific monitoring, and output from a source dispersion model for traffic related air pollution (Caline3QHCR). Accuracy in predicting long-term average concentrations is evaluated using an elaborate cross-validation setup that accounts for a sparse spatio-temporal sampling pattern in the data, and adjusts for temporal effects. The predictive ability of the model is good with cross-validated \(R^2\) of approximately \(0.7\) at subject sites. Replacing four geographic covariate indicators of traffic density with the Caline3QHCR dispersion model output resulted in very similar prediction accuracy from a more parsimonious and more interpretable model. Adding traffic-related geographic covariates to the model that included Caline3QHCR did not further improve the prediction accuracy.