Droplet digital PCR is a cost-effective method for analyzing long cell-free DNA in maternal plasma: Application in preeclampsia

Objective

Long cell-free DNA (cfDNA) can be found in the plasma of pregnant women and cancer patients. We investigated if droplet digital PCR (ddPCR) can analyze such molecules for diagnostic purposes using preeclampsia as a model.

Method

Plasma samples from ten preeclamptic and sixteen normal pregnancies were analyzed. Two ddPCR assays targeting a single-copy gene, VCP, and one ddPCR assay targeting LINE-1 repetitive regions were used to measure the percentages of long cfDNA >533, 1001, and 170 bp, respectively. The LINE-1 assay was developed as guided by in silico PCR analyses to better differentiate preeclamptic and normal pregnancies.

Results

Preeclamptic patients had a significantly lower median percentage of long cfDNA than healthy pregnant controls, as determined by the LINE-1 170 bp assay (28.9% vs. 35.1%, p < 0.0001) and the VCP 533 bp assay (6.6% vs. 8.7%, p = 0.014). The LINE-1 assay provided a better differentiation than the VCP 533 bp assay (area under ROC curves, 0.94 vs. 0.79).

Conclusion

ddPCR is a cost-effective approach for unlocking diagnostic information carried by long cfDNA in plasma and may have applications for the detection of preeclampsia. Further longitudinal studies with larger cohorts are required to assess the clinical utility of this test.     

Prenatal diagnosis of PORCN-related developmental syndrome in a fetus: A novel phenotype

We report a case of a female fetus born to an unrelated couple with a complex fetal phenotype of a pleural effusion, a cardiac malformation, and syndactyly of the toes. Prenatal exome sequencing identified a variant of uncertain significance in the PORCN gene that was upgraded to likely pathogenic following postnatal clinical examination. The phenotype described in cases with variants in the PORCN gene is often associated with findings that cannot be prospectively diagnosed by ultrasonography. This is the first report of a prenatal phenotype involving a fetal effusion associated with variants in the PORCN gene, with skeletal findings identified later in gestation on ultrasonography. The diagnosis was confirmed on neonatal examination.     

Establishing Relevant Ozone Standards to Protect Vegetation and Human Health: Exposure/Dose-Response Considerations

For assessing the efficacy of a specific form of the National Ambient Air Quality Standard for 0₃, those exposure patterns that result in vegetation and human health effects must be identified. For vegetation, it has been found that the higher hourly average concentrations should be weighted more than the lower concentrations. Controlled human exposure work supports the suggestion that concentration may be more important than exposure duration and ventilation rates. It has been indicated in the literature that the current form of the federal 0₃ standard may not be appropriate for protecting vegetation and human health from 0₃ exposures. The proposed use of the cumulative index alone as a form of the standard may not provide sufficient protection to vegetation. An extended-period average index, such as a daily maximum 8-hour average concentration, may not be appropriate to protect human health because of the reduced ability to observe differences among hourly 0₃ concentrations exhibited within exposure regimes. For both vegetation and human health effects research, additional experimentation is required to identify differences in responses that occur when ambient-type exposure regimes are applied. Any standard promulgated to protect vegetation and human health from 0₃ exposures should consider combining cumulative exposure indices with other parameters so that those unique exposures that have the potential for eliciting an adverse effect can be adequately described.     

Size-dependent photosynthetic performance in the giant clam <Emphasis Type="Italic">Tridacna maxima</Emphasis>, a mixotrophic marine bivalve

Giant clams form a symbiosis with photosynthetic algae of the genus Symbiodinium that reside in clam mantle tissue. The allometry of symbiont photosynthetic performance was investigated as a mechanism for the increasing percentage of giant clam carbon respiratory requirements provided by symbionts as clam size increases. Chlorophyll fluorescence measurements of symbionts of the giant clam Tridacna maxima were measured during experiments conducted in September of 2009 using specimens 0.5–200 g tissue wet weight (3–25 cm long), collected from waters around southern Taiwan (N 21°36′, E 120°47′) from July to August of 2009. Light-dependent decreases in effective quantum yield (∆F/F _m′) calculated as the noontime maximum excitation pressure over PSII (Q _m), relative electron transport rates (rETR), and dark-adapted maximum quantum yield (F _v/F _m) all varied as a quadratic function of clam size. Both Q _m and rETR increased as clam size increased up to ~10–50 g then decreased as clam size increased. F _v/F _m decreased as clam size increased up to ~5–50 g then increased as clam size increased. Chlorophyll fluorescence measurements of rETR were positively correlated with gross primary production measured during chamber incubations. Overall, symbionts of mid-sized clams ~5–50 g exhibited the highest light-dependent decreases in effective photosynthetic efficiencies, the highest relative electron transport rates, and the lowest maximum photosynthetic efficiencies, and symbiont photosynthetic performance is allometric with respect to host clam size.     

Empirical examination of the individual‐level personality‐based theory of self‐management failure

The individual‐level personality‐based theory of self‐management failure posits that personality predisposes individuals to self‐defeating behavior that, in turn, leads to self‐management failure (Renn, Allen, Fedor, & Davis, 2005). To provide a partial test of the theory, a model is hypothesized that operationalized personality with neuroticism and conscientiousness of the Big Five personality dimensions; self‐defeating behavior with inability to delay gratification, procrastination, and emotional self‐absorption; and self‐management with personal goal setting, monitoring, and operating. The model was tested using data collected from 286 working employees and structural equations analysis. Results supported nine of 11 theory‐derived hypotheses. As hypothesized, high neuroticism was associated with improper personal goal setting, monitoring, and operating; and emotional self‐absorption and procrastination accounted for the relationship between high neuroticism and ineffective self‐management. In addition, low conscientiousness was associated with inferior self‐management practices, and inability to delay gratification and procrastination partially explained the relationship between low conscientiousness and poor self‐management. The findings provide new insight into how high neuroticism and low conscientiousness may contribute to self‐management failure. Theoretical and practical implications of the study are discussed. Copyright © 2009 John Wiley & Sons, Ltd.     

Plant extract reduces tobacco smoke harmful effects on alveolar macrophage immune responses

Tobacco smoke is a major factor responsible for lung cancer and chronic obstructive pulmonary disease. Although the best solution to reduce the incidence of these diseases is to quit smoking, there are still a large number of smokers. Thus, given the immunoregulatory properties of plant extracts, their capacity to reduce tobacco smoke harmful effects on alveolar macrophage (AM) functions was investigated. AM were treated with tobacco smoke extract and parenchymata tissue extract (PTE), or mesophyll cell extract (MCE) of Spinacia oleacea. The effects of tobacco smoke extract from PTE and MCE-treated cigarette filters were also investigated. AM production of tumor necrosis factor (TNF), interleukin-10 (IL-10) and macrophage chemoattractant protein-1 (MCP-1), and AM cytotoxicity were measured. Tobacco smoke extract significantly inhibited TNF, IL-10, and MCP-1 release, and AM cytotoxicity. The addition of PTE and MCE to tobacco smoke extract abrogated the inhibition of AM mediator release. However, only MCE restored AM cytotoxicity. Interestingly, tobacco smoke extract of PTE and MCE-treated cigarette filters showed reduced effects on AM functions. Tobacco smoke extract from MCE-treated (0.25%) cigarette filters did not inhibit TNF, IL-10, and MCP-1 release in contrast to tobacco smoke extract from buffer-treated cigarette filters. AM cytotoxic activity was not inhibited by the treatment with tobacco smoke extract from MCE-treated cigarette filters. Our data suggest that the presence of plant extract in cigarette filters reduces the inhibitory effects of cigarette smoke on AM functions. Thus, MCE-treated cigarette filters may help reducing lung diseases associated with smoking.     

Species, functional groups, and thresholds in ecological resilience

The cross-scale resilience model states that ecological resilience is generated in part from the distribution of functions within and across scales in a system. Resilience is a measure of a system's ability to remain organized around a particular set of mutually reinforcing processes and structures, known as a regime. We define scale as the geographic extent over which a process operates and the frequency with which a process occurs. Species can be categorized into functional groups that are a link between ecosystem processes and structures and ecological resilience. We applied the cross-scale resilience model to avian species in a grassland ecosystem. A species' morphology is shaped in part by its interaction with ecological structure and pattern, so animal body mass reflects the spatial and temporal distribution of resources. We used the log-transformed rank-ordered body masses of breeding birds associated with grasslands to identify aggregations and discontinuities in the distribution of those body masses. We assessed cross-scale resilience on the basis of 3 metrics: overall number of functional groups, number of functional groups within an aggregation, and the redundancy of functional groups across aggregations. We assessed how the loss of threatened species would affect cross-scale resilience by removing threatened species from the data set and recalculating values of the 3 metrics. We also determined whether more function was retained than expected after the loss of threatened species by comparing observed loss with simulated random loss in a Monte Carlo process. The observed distribution of function compared with the random simulated loss of function indicated that more functionality in the observed data set was retained than expected. On the basis of our results, we believe an ecosystem with a full complement of species can sustain considerable species losses without affecting the distribution of functions within and across aggregations, although ecological resilience is reduced. We propose that the mechanisms responsible for shaping discontinuous distributions of body mass and the nonrandom distribution of functions may also shape species losses such that local extinctions will be nonrandom with respect to the retention and distribution of functions and that the distribution of function within and across aggregations will be conserved despite extinctions.     

Identification of the putative aggregation pheromone components emitted by the European earwig, <Emphasis Type="Italic">Forficula auricularia</Emphasis>

The European earwig, Forficula auricularia is an invasive insect found in many temperate regions of the world and is regarded as an urban and agricultural pest of numerous crops. Several studies have shown that F. auricularia aggregate in large numbers with the use of an aggregation pheromone. However, the identity of compounds which constitute the pheromone remains elusive. Our aim was to isolate and identify the aggregation pheromone used by F. auricularia using both solvent washes of individuals and thermal desorption of substrates exposed to earwigs. Solvent washes of male, female and juvenile earwigs isolated 51 different branched and unbranched alkanes, alkenes and alkadienes. Substrates exposed to aggregating field populations in situ were demonstrated to be attractive to earwigs after less than 24 h of exposure. Analysis of these substrates using thermal desorption and solvent washes showed that hydrocarbons were the only detectable compounds laid down by earwigs on these surfaces. Significant behavioural responses were observed to synthetic blends of the unsaturated hydrocarbons containing (Z)-7-tricosene, (Z)-9-tricosene, (Z)-7-pentacosene and (Z)-9-pentacosene at ≥25 insect equivalents in field-based bioassays. However, behavioural responses to these blends proved inconsistent particularly later in the field season, possibly due to a missing component within the pheromone blend or plasticity in the pheromones production and response.