Mid‐term effects on ecosystem services of quarry restoration with Technosols under Mediterranean conditions: 10‐year impacts on soil organic carbon and vegetation development

The use of Technosols for the restoration of limestone quarries overcomes the usual “in situ” scarcity of soil and/or its poor quality. The use of mine spoils, improved with mineral and/or organic amendments, could be an efficient and environmentally friendly option. Properly treated sewage sludge from urban wastewater treatment plants could be a suitable organic amendment and fertilizer (rich in N and P) whenever its pollutant burden is low (heavy metals and/or organic pollutants). Its appropriate use could improve essential soil physical and chemical properties and, therefore, promote key ecosystem services of restored areas, such as biomass production and carbon sequestration, as well as biodiversity and landscape recovery. However, the mid‐term impacts of these restoration practices on soil functioning and their services have rarely been reported in the available literature. In this study we assess the mid‐term effects (10 years) of the use of sewage sludge as a Technosol amendment on soil organic carbon (SOC), nutrient status, and plant development in several restored quarries. Soils restored using sewage sludge showed a threefold increase in SOC compared to the corresponding unamended ones, despite the moderate sludge dosage applied (below 50 tonnes/ha). Plant cover was also higher in amended soils, and recruitment was not affected by sludge amendment at these doses. This study demonstrates that, used at an appropriate rate, sewage sludge is a good alternative for the valorization of mine spoils in quarry restoration, improving some important regulatory ecosystem services such as carbon sequestration, without compromising woody plant encroachment.     

In vitro antimicrobial activity of plant extracts against Pseudomonas syringae pv. actinidiae causal agent of bacterial canker in kiwifruit

Abstract

Pseudomonas syringae pv. actinidiae (Psa), the causal agent of bacterial canker of kiwifruit, is considered the main pathogen of yellow-, green- and red-fleshed kiwifruit. All major kiwifruit producing countries in the world have been affected by this bacterial pathogen, leading to substantial economic losses. The control of bacterial canker of kiwifruit is based only on preventive methods or on the use of copper compounds that can cause phytotoxicity problems. In this study, the in vitro antibacterial activity of seven different plant extracts against eight Psa strains has been evaluated. The inhibition of 100% of the Psa growth was observed, after 24?h, for the extracts of Polygonum cuspidatum roots (POL-roots), Hypericum perforatum roots elicited with chitosan oligosaccharides (HYP-COS roots) and non-fermented grape pomace (ITA-pomace). The strongest antibacterial activity was exhibited by POL-roots, with a geometric mean of minimum inhibitory concentration of 100% of growth (GMMIC₁₀₀) of 105.11 µg/mL after 24?h, and with a GMMIC₁₀₀ value of 148.65 µg/mL after 48?h. Moreover, POL-roots extract showed the best bactericidal activity with a GMMBC of 210.22 µg/mL. No phytotoxic activity was observed up to 15 days in the leaves of Actinidia chinensis “Belen” treated with plant extracts at 500 µg/mL.     

Co-Registering Kinematics and Evoked Related Potentials during Visually Guided Reach-to-Grasp Movements

Background

In non-human primates grasp-related sensorimotor transformations are accomplished in a circuit involving the anterior intraparietal sulcus (area AIP) and both the ventral and the dorsal sectors of the premotor cortex (vPMC and dPMC, respectively). Although a human homologue of such a circuit has been identified, the time course of activation of these cortical areas and how such activity relates to specific kinematic events has yet to be investigated.

Methodology/Principal Findings

We combined kinematic and event-related potential techniques to explicitly test how activity within human grasping-related brain areas is modulated in time. Subjects were requested to reach towards and grasp either a small stimulus using a precision grip (i.e., the opposition of index finger and thumb) or a large stimulus using a whole hand grasp (i.e., the flexion of all digits around the stimulus). Results revealed a time course of activation starting at the level of parietal regions and continuing at the level of premotor regions. More specifically, we show that activity within these regions was tuned for specific grasps well before movement onset and this early tuning was carried over - as evidenced by kinematic analysis - during the preshaping period of the task.

Conclusions/Significance

Data are discussed in terms of recent findings showing a marked differentiation across different grasps during premovement phases which was carried over into subsequent movement phases. These findings offer a substantial contribution to the current debate about the nature of the sensorimotor transformations underlying grasping. And provide new insights into the detailed movement information contained in the human preparatory activity for specific hand movements.     

Bone Mineral Density,Bone Turnover Markers and Fractures in Patients with Systemic Sclerosis: A Case Control Study

Objective

The aim of our study was to elucidate the pathophysiology of systemic sclerosis-related osteoporosis and the prevalence of vertebral fragility fracture in postmenopausal women with systemic sclerosis (SSc).

Methodology

Fifty-four postmenopausal women with scleroderma and 54 postmenopausal controls matched for age, BMI, and smoking habits were studied. BMD was measured by dual energy-x-ray absorptiometry at spine and femur, and by ultrasonography at calcaneus The markers of bone turnover included serum osteocalcin and urinary deoxypyridinoline. All subjects had a spine X-ray to ascertain the presence of vertebral fractures.

Results

bone mineral density at lumbar spine (BMD 0.78±0.08 vs 0.88±0.07; p<0,001), femoral neck (BMD: 0.56±0.04 vs 0.72±0.07; p<0,001) and total femur (BMD: 0.57±0.04 vs 0.71±0.06; p<0,001) and ultrasound parameter at calcaneus (SI: 80.10±5.10 vs 94.80±6.10 p<0,001) were significantly lower in scleroderma compared with controls; bone turnover markers and parathyroid hormone level were significantly higher in scleroderma compared with controls, while serum of 25(OH)D3 was significantly lower. In scleroderma group the serum levels of 25(OH)D3 significantly correlated with PTH levels, BMD, stiffness index and bone turnover markers. One or more moderate or severe vertebral fractures were found in 13 patients with scleroderma, wherease in control group only one patient had a mild vertebral fracture.

Conclusion

Our data shows, for the first time, that vertebral fractures are frequent in subjects with scleroderma, and suggest that lower levels of 25(OH)D3 may play a role in the risk of osteoporosis and vertebral fractures.     

Novel Targets of Sulforaphane in Primary Cardiomyocytes Identified by Proteomic Analysis

Cardiovascular diseases represent the main cause of mortality in the industrialized world and the identification of effective preventive strategies is of fundamental importance. Sulforaphane, an isothiocyanate from cruciferous vegetables, has been shown to up-regulate phase II enzymes in cardiomyocytes and counteract oxidative stress-induced apoptosis. Aim of the present study was the identification and characterization of novel sulforaphane targets in cardiomyocytes applying a proteomic approach. Two-dimensional gel electrophoresis and mass spectrometry were used to generate protein profiles of primary neonatal rat cardiomyocytes treated and untreated with 5 µM sulforaphane for 1-48 h. According to image analysis, 64 protein spots were found as differentially expressed and their functional correlations were investigated using the MetaCore program. We mainly focused on 3 proteins: macrophage migration inhibitory factor (MIF), CLP36 or Elfin, and glyoxalase 1, due to their possible involvement in cardioprotection. Validation of the time-dependent differential expression of these proteins was performed by western blotting. In particular, to gain insight into the cardioprotective role of the modulation of glyoxalase 1 by sulforaphane, further experiments were performed using methylglyoxal to mimic glycative stress. Sulforaphane was able to counteract methylglyoxal-induced apoptosis, ROS production, and glycative stress, likely through glyoxalase 1 up-regulation. In this study, we reported for the first time new molecular targets of sulforaphane, such as MIF, CLP36 and glyoxalase 1. In particular, we gave new insights into the anti-glycative role of sulforaphane in cardiomyocytes, confirming its pleiotropic behavior in counteracting cardiovascular diseases.     

Imaging of the DNA damage‐induced dynamics of nuclear proteins via nonlinear photoperturbation

Understanding the cellular response to DNA strand breaks is crucial to decipher the mechanisms maintaining the integrity of our genome. We present a novel method to visualize how the mobility of nuclear proteins changes in response to localized DNA damage. DNA strand breaks are induced via nonlinear excitation with femtosecond laser pulses at λ = 1050 nm in a 3D‐confined subnuclear volume. After a time delay of choice, protein mobility within this volume is analysed by two‐photon photoactivation of PA‐GFP fusion proteins at λ = 775 nm. By changing the position of the photoactivation spot with respect to the zone of lesion the influence of chromatin structure and of the distance from damage are investigated. As first applications we demonstrate a locally confined, time‐dependent mobility increase of histone H1.2, and a progressive retardation of the DNA repair factor XRCC1 at damaged sites. This assay can be used to map the response of nuclear proteins to DNA damage in time and space. (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Environmental Conditioning of Skeletal Anomalies Typology and Frequency in Gilthead Seabream (Sparus aurata L., 1758) Juveniles

In this paper, 981 reared juveniles of gilthead seabream (Sparus aurata) were analysed, 721 of which were from a commercial hatchery located in Northern Italy (Venice, Italy) and 260 from the Hellenic Center for Marine Research (Crete, Greece). These individuals were from 4 different egg batches, for a total of 10 different lots. Each egg batch was split into two lots after hatching, and reared with two different methodologies: intensive and semi-intensive. All fish were subjected to processing for skeletal anomaly and meristic count analysis. The aims involved: (1) quantitatively and qualitatively analyzing whether differences in skeletal elements arise between siblings and, if so, what they are; (2) investigating if any skeletal bone tissue/ossification is specifically affected by changing environmental rearing conditions; and (3) contributing to the identification of the best practices for gilthead seabream larval rearing in order to lower the deformity rates, without selections. The results obtained in this study highlighted that: i) in all the semi-intensive lots, the bones having intramembranous ossification showed a consistently lower incidence of anomalies; ii) the same clear pattern was not observed in the skeletal elements whose ossification process requires a cartilaginous precursor. It is thus possible to ameliorate the morphological quality (by reducing the incidence of severe skeletal anomalies and the variability in meristic counts of dermal bones) of reared seabream juveniles by lowering the stocking densities (maximum 16 larvae/L) and increasing the volume of the hatchery rearing tanks (minimum 40 m³). Feeding larvae with a wide variety of live (wild) preys seems further to improve juvenile skeletal quality. Additionally, analysis of the morphological quality of juveniles reared under two different semi-intensive conditions, Mesocosm and Large Volumes, highlighted a somewhat greater capacity of Large Volumes to significantly augment the gap with siblings reared in intensive (conventional) modality.     

Parental Imbalances Involving Chromosomes 15q and 22q May Predispose to the Formation of De Novo Pathogenic Microdeletions and Microduplications in the Offspring

Microarray-based comparative genomic hybridization (array-CGH) led to the discovery of genetic abnormalities among patients with complex phenotype and normal karyotype. Also several apparently normal individuals have been found to be carriers of cryptic imbalances, hence the importance to perform parental investigations after the identification of a deletion/duplication in a proband. Here, we report the molecular cytogenetic characterization of two individuals in which the microdeletions/duplications present in their parents could have predisposed and facilitated the formation of de novo pathogenic different copy number variations (CNVs). In family 1, a 4-year-old girl had a de novo pathogenic 10.5 Mb duplication at 15q21.2q22.2, while her mother showed a 2.262 Mb deletion at 15q13.2q13.3; in family 2, a 9-year-old boy had a de novo 1.417 Mb deletion at 22q11.21 and a second paternal deletion of 247 Kb at 22q11.23 on the same chromosome 22. Chromosome 22 at band q11.2 and chromosome 15 at band q11q13 are considered unstable regions. We could hypothesize that 15q13.2q13.3 and 22q11.21 deletions in the two respective parents might have increased the risk of rearrangements in their children. This study highlights the difficulty to make genetic counseling and predict the phenotypic consequences in these situations.     

A Plant Virus Manipulates the Behavior of Its Whitefly Vector to Enhance Its Transmission Efficiency and Spread

Plant viruses can produce direct and plant-mediated indirect effects on their insect vectors, modifying their life cycle, fitness and behavior. Viruses may benefit from such changes leading to enhanced transmission efficiency and spread. In our study, female adults of Bemisia tabaci were subjected to an acquisition access period of 72 h in Tomato yellow leaf curl virus (TYLCV)-infected and non-infected tomato plants to obtain viruliferous and non-viruliferous whiteflies, respectively. Insects that were exposed to virus-infected plants were checked by PCR to verify their viruliferous status. Results of the Ethovision video tracking bioassays indicated that TYLCV induced an arrestant behavior of B. tabaci, as viruliferous whitefly adults remained motionless for more time and moved slower than non-viruliferous whiteflies after their first contact with eggplant leaf discs. In fact, Electrical Penetration Graphs showed that TYLCV-viruliferous B. tabaci fed more often from phloem sieve elements and made a larger number of phloem contacts (increased number of E1, E2 and sustained E2 per insect, p<0.05) in eggplants than non-viruliferous whiteflies. Furthermore, the duration of the salivation phase in phloem sieve elements (E1) preceding sustained sap ingestion was longer in viruliferous than in non-viruliferous whiteflies (p<0.05). This particular probing behavior is known to significantly enhance the inoculation efficiency of TYLCV by B. tabaci. Our results show evidence that TYLCV directly manipulates the settling, probing and feeding behavior of its vector B. tabaci in a way that enhances virus transmission efficiency and spread. Furthermore, TYLCV-B. tabaci interactions are mutually beneficial to both the virus and its vector because B. tabaci feeds more efficiently after acquisition of TYLCV. This outcome has clear implications in the epidemiology and management of the TYLCV-B. tabaci complex.