The utility of DNA barcoding as a tool to assess the success of ecological restoration using Hemiptera as a biological indicator

DNA barcoding uses a short, standardized DNA fragment to sort individuals into species. This molecular technique has applications in fields including ecology, evolution, conservation, and biogeography. In ecological applications such as species monitoring and habitat restoration, its potential has not been fully realized and implemented. Invertebrates are excellent biological indicators, as changes in species diversity or community assemblage provide important insights into the condition of, or changes in, the environment. This information is particularly useful within the context of restoration ecology. In this study, DNA barcoding is used to assess the potential of Hemiptera as a biological indicator of restoration success for the Buffelsdraai Landfill Site Community Reforestation Project (Durban, South Africa). A total of 393 Hemiptera specimens were collected from sites reforested at distinct phases (plots reforested in 2010, 2012, and 2015) and two reference sites (natural forest and grassland). The Hemiptera species composition and assemblage were assessed by analyzing diversity indices, ordination, unweighted pair‐group average cluster analysis, and phylogenetic analysis. Hemiptera species composition varied significantly across the chronologically different reforested sites, with a higher species richness observed in the older reforested plots. This suggests that Hemiptera diversity can be used to track restoration success, even over the small temporal scale used in this study. This study highlights the utility of DNA barcoding as a taxonomic sorting tool both to monitor ecological restoration and to discover specific taxa within Hemiptera that may be useful biological indicators.     

Utility of nuclear DNA intron markers at lower taxonomic levels: phylogenetic resolution among nine Tragelaphus spp

Phylogenetic relationships among the nine spiral-horn antelope species of the African bovid tribe Tragelaphini are controversial. In particular, mitochondrial DNA sequencing studies are not congruent with previous morphological investigations. To test the utility of nuclear DNA intron markers at lower taxonomic levels and to provide additional data pertinent to tragelaphid evolution, we sequenced four nuclear DNA segments (MGF, PRKCI, SPTBN, and THY) and combined these data with mitochondrial DNA sequences from three genes (cytochrome b, 12S rRNA, and 16S rRNA). Our molecular supermatrix comprised 4682 characters which were analyzed independently and in combination. Parsimony and model based phylogenetic analyses of the combined nuclear DNA data are congruent with those derived from the analysis of mitochondrial gene sequences. The corroboration between nuclear and mtDNA gene trees reject the possibility that genetic processes such as lineage sorting, gene duplication/deletion and hybrid speciation account for the conflict evident in the previously published phylogenies. It suggests rather that the morphological characters used to delimit the Tragelaphid species are subject to convergent evolution. Divergence times among species, calculated using a relaxed Bayesian molecular clock, are consistent with hypotheses proposing that climatic oscillations and their impact on habitats were the major forces driving speciation in the tribe Tragelaphini.     

Statistical Parameters in Behavioral Tasks and Implications for Sample Size of C57BL/6J:129S6/SvEvTac Mixed Strain Mice

Most mixed strain progeny from gene-knockout experiments typically originate from C57BL/6J and one of the 129 substrains, frequently 129S6/SvEvTac. The results of this behavioral survey suggest that C57BL/6J:129S6/SvEvTac mixed strain mice are amenable to behavioral testing. The variability in behavioral tasks for subjects arising from this mixed strain genetic background does not preclude screening with a battery of behavioral tests. With clues provided by a screen of mixed strain subjects, follow-up analyses with isogenic, congenic, or F1 hybrid animals may be targeted to specific behavioral themes.     

Adjustment of the PIF7‐HFR1 transcriptional module activity controls plant shade adaptation

Shade caused by the proximity of neighboring vegetation triggers a set of acclimation responses to either avoid or tolerate shade. Comparative analyses between the shade‐avoider Arabidopsis thaliana and the shade‐tolerant Cardamine hirsuta revealed a role for the atypical basic‐helix‐loop‐helix LONG HYPOCOTYL IN FR 1 (HFR1) in maintaining the shade tolerance in C. hirsuta, inhibiting hypocotyl elongation in shade and constraining expression profile of shade‐induced genes. We showed that C. hirsuta HFR1 protein is more stable than its A. thaliana counterpart, likely due to its lower binding affinity to CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1), contributing to enhance its biological activity. The enhanced HFR1 total activity is accompanied by an attenuated PHYTOCHROME INTERACTING FACTOR (PIF) activity in C. hirsuta. As a result, the PIF‐HFR1 module is differently balanced, causing a reduced PIF activity and attenuating other PIF‐mediated responses such as warm temperature‐induced hypocotyl elongation (thermomorphogenesis) and dark‐induced senescence. By this mechanism and that of the already‐known of phytochrome A photoreceptor, plants might ensure to properly adapt and thrive in habitats with disparate light amounts.     

Cellular,Molecular and Functional Characterisation of YAC Transgenic Mouse Models of Friedreich Ataxia

Background

Friedreich ataxia (FRDA) is an autosomal recessive neurodegenerative disorder, caused by a GAA repeat expansion mutation within intron 1 of the FXN gene. We have previously established and performed preliminary characterisation of several human FXN yeast artificial chromosome (YAC) transgenic FRDA mouse models containing GAA repeat expansions, Y47R (9 GAA repeats), YG8R (90 and 190 GAA repeats) and YG22R (190 GAA repeats).

Methodology/Principal Findings

We now report extended cellular, molecular and functional characterisation of these FXN YAC transgenic mouse models. FXN transgene copy number analysis of the FRDA mice demonstrated that the YG22R and Y47R lines each have a single copy of the FXN transgene while the YG8R line has two copies. Single integration sites of all transgenes were confirmed by fluorescence in situ hybridisation (FISH) analysis of metaphase and interphase chromosomes. We identified significant functional deficits, together with a degree of glucose intolerance and insulin hypersensitivity, in YG8R and YG22R FRDA mice compared to Y47R and wild-type control mice. We also confirmed increased somatic GAA repeat instability in the cerebellum and brain of YG22R and YG8R mice, together with significantly reduced levels of FXN mRNA and protein in the brain and liver of YG8R and YG22R compared to Y47R.

Conclusions/Significance

Together these studies provide a detailed characterisation of our GAA repeat expansion-based YAC transgenic FRDA mouse models that will help investigations of FRDA disease mechanisms and therapy.     

Diversity and oenological characterization of indigenous <Emphasis Type="Italic">Saccharomyces</Emphasis><Emphasis Type="Italic">cerevisiae</Emphasis> associated with Žilavka grapes

The aim of this research was to identify the Saccharomyces spp. associated with Žilavka grapes and to evaluate their enzymatic activities, H₂S production and micro-fermentation performance. For this purpose, a total of 143 yeast strains isolated from three production areas of the Mostar wine region (Bosnia and Herzegovina) were studied and analysed. Firstly, yeasts were identified to genus level by growth on WL nutrient agar and the test of assimilation of lysine. Later, molecular identification at species level was carried out with RFLP analysis of 18S rDNA + ITS region, and at strain level with microsatellite-primed PCR (MSP-PCR). At physiological level yeast strains were grouped into different clusters by means of the Joining-Tree-Clustering-Method. All yeasts tested were identified as S. cerevisiae, resulting a total of 18 different strains. All of the investigated strains produced hydrogen sulphide, 89% were able to complete the fermentation, and none of them was able to synthesize killer toxins. Since enzymes play a very important role in wine aroma development, it was very encouraging that 33% of the strains were able to synthesize pectinolytic enzyme but only one produced β-glucosidase. In the second part of the selection process two indigenous strains were compared with commercial yeast in a microvinification and Žilavka wines with different profiles of volatiles were obtained. This research represents a first step in the selection of indigenous yeast strains from the Mostar region with the goal of maintaining the specific organoleptic characteristics of Žilavka wine.     

Differential impact of polysialyltransferase ST8SiaII and ST8SiaIV knockout on social interaction and aggression

Previous studies using neuronal cell adhesion molecule (NCAM) ?/? knockout (KO) mice provided evidence for a role of NCAMs in social behaviors. However, polysialic acid (PSA), the most important post‐translational modification of NCAM, was also absent in these mice, which makes it difficult to distinguish between the specific involvement of either PSA or NCAM in social interactions. To address this issue, we assessed two lines of mice deficient for one of the two sialyltransferase enzymes required for the polysialylation of NCAM, sialyltransferase‐X (St8SiaII or STX) and polysialyltransferase (ST8SiaIV or PST), in a series of tests for social behaviors. Results showed that PST KO mice display a decreased motivation in social interaction. This deficit can be partly explained by olfactory deficits and was associated with a clear decrease in PSA‐NCAM expression in all brain regions analyzed (amygdala, septum, bed nucleus of the stria terminalis and frontal cortices). STX KO mice displayed both a decreased social motivation and an increased aggressive behavior that cannot be explained by olfactory deficits. This finding might be related to the reduced anxiety‐like behavior, increased locomotion and stress‐induced corticosterone secretion observed in these mice. Moreover, STX KO mice showed mild increase of PSA‐NCAM expression in the lateral septum and the orbitofrontal cortex. Altogether, these findings support a role for PSA‐NCAM in the regulation of social behaviors ranging from a lack of social motivation to aggression. They also underscore STX KO mice as an interesting animal model that combines a behavioral profile of violence and hyperactivity with reduced anxiety‐like behavior.     

Whispered voice test for screening for hearing impairment in adults and children: systematic review

Objective To determine the accuracy of the whispered voice test in detecting hearing impairment in adults and children.Design Systematic review of studies of test accuracy.Data sources Medline, Embase, Science Citation Index, unpublished theses, manual searching of bibliographies of known primary and review articles, and contact with authors.Study selection Two reviewers independently selected and extracted data on study characteristics, quality, and accuracy of studies. Studies were included if they had cross sectional designs, at least one of the index tests was the whispered voice test, and the reference test (audiometry) was performed on at least 80% of the participants.Data extraction Data were used to form 2×2 contingency tables with hearing impairment by audiometry as the reference standard.Data synthesis The eight studies that were found used six different techniques. The sensitivity in the four adult studies was 90% or 100% and the specificity was 70% to 87%. The sensitivity in the four childhood studies ranged from 80% to 96% and specificity ranged from 90% to 98%.Conclusion The whispered voice test is a simple and accurate test for detecting hearing impairment. There is some concern regarding the lower sensitivity in children and the overall reproducibility of the test, particularly in primary care settings. Further studies should be conducted in primary care settings to explore the influence of components of the testing procedure to optimise test sensitivity and to promote standardisation of the testing procedure.