Proteomic Screening of Antigenic Proteins from the Hard Tick,Haemaphysalis longicornis (Acari: Ixodidae)

Proteomic tools allow large-scale, high-throughput analyses for the detection, identification, and functional investigation of proteome. For detection of antigens from Haemaphysalis longicornis, 1-dimensional electrophoresis (1-DE) quantitative immunoblotting technique combined with 2-dimensional electrophoresis (2-DE) immunoblotting was used for whole body proteins from unfed and partially fed female ticks. Reactivity bands and 2-DE immunoblotting were performed following 2-DE electrophoresis to identify protein spots. The proteome of the partially fed female had a larger number of lower molecular weight proteins than that of the unfed female tick. The total number of detected spots was 818 for unfed and 670 for partially fed female ticks. The 2-DE immunoblotting identified 10 antigenic spots from unfed females and 8 antigenic spots from partially fed females. Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF) of relevant spots identified calreticulin, putative secreted WC salivary protein, and a conserved hypothetical protein from the National Center for Biotechnology Information and Swiss Prot protein sequence databases. These findings indicate that most of the whole body components of these ticks are non-immunogenic. The data reported here will provide guidance in the identification of antigenic proteins to prevent infestation and diseases transmitted by H. longicornis.     

Antioxidant,antimicrobial, toxicity and analgesic properties of ethanol extract of Solena amplexicaulis root

Background

This study was subjected to investigate different pharmacological properties of ethanol extract of Solena amplexicaulis root.

Results

The extract contains flavonoid, alkaloid, saponin and steroid compounds. The extract exhibited excellent antioxidant activity in DPPH radical scavenging activity. The extract also showed potent activity in brine shrimp lethality bioassay. The LC₅₀ value was found to 44.677 μg/ml. The extract showed better anti-bacterial activity against gram-negative bacteria. In antifungal assay, the maximum 79.31% of anti-mycotic activity was observed against Aspergillus ochraceus while minimum 44.2% against Rhizopus oryzae. MIC value ranged between 1500–3000 μg/ml. The extract was found moderately toxic with a 24-hr LD₅₀ value of 81.47 mg/kg in Swiss albino mice. The degree of inhibition by the ethanolic extract of the root was found less than that of standard analgesic drug diclofenac sodium. The extract also showed moderate anti-inflammatory and antinociceptive activity and anti-diabetic property. Reducing power of the extract was comparable with standard ascorbic acid. Moderate in vitro thrombolytic activity, lipid peroxidation inhibition property, metal chelating ability and stress-protective activity was also observed.

Conclusion

Ethanol extract of Solena amplexicaulis root can be valuable for treatment of different diseases.     

Invaders' control on post-disturbance succession in coastal mangroves

Aims In recent years, coastal mangroves have been frequently affected by large disturbances (cyclones, hurricanes, flooding and tsunamis) and post-disturbance vegetation is often dominated by small stature mangrove, mangrove-associate and non-mangrove species potentially affecting ecosystem functioning. Knowledge on the processes of mangrove vegetation development and recovery (succession) following normal and large disturbances will benefit practitioners in designing robust ecosystem management/restoration plans. Here we propose a conceptual model of disturbance-mediated succession in mangroves.Methods Based on field observations and species' life history traits, we develop conceptual models of mangrove succession under normal disturbance regime and recently experienced increased frequency of large disturbances. We evaluate our conceptual models by conducting a scenario testing experiment.Important findings We suggest two predominant processes affecting mangrove succession after disturbance: propagule limitation due to damage of seed producing mature trees and dispersal barrier resulting from biological invasion associated with large disturbance. We argue that large disturbances affect mature trees more than the small-stature non-tree (shrubs, herbs and climbers) species creating a larger propagule shortage for mangrove tree species than non-tree species. Secondly, large disturbances facilitate invasion of free-floating aquatics, which may interfere with the flow-facilitated propagule dispersal and seedling establishment of mangrove species. In a scenario testing experiment, we have shown that similar levels of disturbance impact vegetation development and recovery differently depending on the presence or absence of invasive species. We conclude that since biological invasion is one of the major drivers of post-disturbance mangrove succession, the dimension of biological invasion should be included in prediction, management and restoration of mangrove forests.     

Increases in leaf nitrogen concentration and leaf area did not enhance spur survival and return bloom in almonds (<Emphasis Type="Italic">Prunus dulcis</Emphasis> [Mill.] DA Webb)

Mature almond trees bear fruit mainly on short shoots called spurs, with only a small percentage of fruit produced laterally on long 1-year-old shoots. As a result, maintenance of large numbers of healthy spurs per tree is critical for fruit production. However, spurs that bear fruit have lower leaf area, leaf nitrogen content, and CO₂ assimilation rate than non-fruiting spurs. This has been correlated with reduced percentages of spur survival and return bloom the following season. Thus, we hypothesized that spur leaf area, and ultimately spur health could be enhanced through application of foliar sprays and soil nitrogen treatments that would enhance leaf nitrogen content and spur leaf area. To test our hypothesis, we selected almond trees exhibiting significant yield differences as a consequence of differential soil rates of nitrogen fertilization (N rate) for three prior years (140, 224, and 392 kg/ha). In each tree, three spur types [non-fruiting spurs (F0); spurs with one fruit (F1); spurs with two fruit (F2)] were selected on the east side of the canopy and tracked for one complete season (2011–2012). Four foliar treatments (nutrient replacement, nutrient replacement with biostimulant, nitrogen, and non-spray) were directly applied to individual spurs in each N rate in the spring of 2011 and characteristic such as leaf nitrogen, and fruit quality were recorded throughout the season. In winter of 2012, spur survival and return bloom were addressed through individual visual inspection of the tagged spurs. In this experiment, soil and foliar N treatments effectively increased spur leaf area, fruit, and leaf nitrogen concentration. In the high N treatment, the leaf nitrogen values exceeded the critical nitrogen concentration established for almond trees and the critical leaf area for spur survival and blooming thresholds established by past research in this area. However, none of these positive changes in leaf N or leaf area improved spur survival and/or return boom of any spur type. Indeed, survival and hull + shell weight tended to be lower in the population of fruiting spurs with the highest leaf area and leaf nitrogen concentration and the return bloom probabilities were always lower in fruiting spurs than in non-fruiting spurs, independent of the nitrogen rate. These results and the relationship between nitrogen rate and spur survival are discussed.     

Carbofuran accelerates the cellular senescence and declines the life span of spns1 mutant zebrafish

Carbofuran is a carbamate pesticide, widely used in agricultural practices to increase crop productivity. In mammals, carbofuran is known to cause several untoward effects, such as apoptosis in the hippocampal neuron, oxidative stress, loss of memory and chromosomal anomalies. Most of these effects are implicated with cellular senescence. Therefore, the present study aimed to determine the effect of carbofuran on cellular senescence and biological ageing. Spinster homolog 1 (Spns1) is a transmembrane transporter, regulates autolysosomal biogenesis and plays a role in cellular senescence and survival. Using senescence-associated β-galactosidase staining, we found that carbofuran accelerates the cellular senescence in spns1 mutant zebrafish. The yolk opaqueness, a premature ageing phenotype in zebrafish embryos, was accelerated by carbofuran treatment. In the survival study, carbofuran shortened the life span of spns1 mutant zebrafish. Autophagy is the cellular lysosomal degradation, usually up-regulated in the senescent cells. To know the impact of carbofuran exposure on autophagy progress, we established a double-transgenic zebrafish line, harbouring EGFP-tagged LC3-II and mCherry-tagged Lamp1 on spns1 mutant background, whereas we found, carbofuran exposure synergistically accelerates autolysosome formation with insufficient lysosome-mediated degradation. Our data collectively suggest that carbofuran exposure synergistically accelerates the cellular senescence and affects biological ageing in spns1 defective animals.     

Effects of anthropogenic disturbance on the species assemblages of birds in the back mangrove forests

Wetlands Ecology and Management - Around the world, mangrove ecosystem has faced major threats from human activities, and birds were unexceptionally impacted. Mangrove fringe a large extent of the...     

Copper complex derived from <Emphasis Type="Italic">S</Emphasis>-benzyldithiocarbazate and 3-acetylcoumarin induced apoptosis in breast cancer cell

Copper complexes have been widely studied for the anti-tumour application as cancer cells are reported to take up greater amounts of copper than normal cells. Preliminary study revealed that the newly synthesised copper complex [Cu(SBCM)₂] displayed marked anti-proliferative towards triple-negative MDA-MB-231 breast cancer cells. Therefore, Cu(SBCM)₂ has great potential to be developed as an agent for the management of breast cancer. The present study was carried out to investigate the mode of cell death induced by Cu(SBCM)₂ towards MDA-MB-231 breast cancer cells. The inhibitory and morphological changes of MDA-MB-231 cells treated with Cu(SBCM)₂ was determined by using MTT assay and inverted light microscope, respectively. The safety profile of Cu(SBCM)₂ was also evaluated towards human dermal fibroblast (HDF) normal cells. Confirmation of apoptosis and cell cycle arrest were determined by flow cytometry analysis. The expression of p53, Bax, Bcl-2 and MMP2 protein were detected with western blot analysis. Cu(SBCM)₂ significantly inhibited the growth of MDA-MB-231 cells in a dose-dependent manner with GI₅₀ 18.7?±?3.06 µM. Indeed, Cu(SBCM)₂ was less toxic towards HDF normal cells with GI₅₀ 31.8?±?4.0 µM. Morphological study revealed that Cu(SBCM)₂-treated MDA-MB-231 cells experienced cellular shrinkage, membrane blebbing, chromatin condensation and formation of apoptotic bodies, suggesting that Cu(SBCM)₂ induced apoptosis in the cells, which was confirmed by Annexin-V/PI flow cytometry analysis. It was also found that Cu(SBCM)₂ induced G₂/M phase cell cycle arrest towards MDA-MB-231 cells. The induction of apoptosis and cell cycle arrest in the present study is possibly due to the down-regulation of the mutant p53 and MMP2 protein. In conclusion, Cu(SBCM)₂ can be developed as a targeted therapy for the treatment of triple-negative breast cancer.     

Prediction of leaf nitrogen from early season samples and development of field sampling protocols for nitrogen management in Almond (Prunus dulcis [Mill.] DA Webb)

Background and aim

Protocols for leaf sampling in deciduous tree crops are commonly executed too late in the season and do not adequately consider field variability to be effectively used to guide N management. The goal of this study was to develop improved sampling strategies to optimize nitrogen management in deciduous tree crops.

Method

Leaf nutrient concentration from individual trees in four mature commercial orchards was collected (n =1148) for three consecutive seasons to develop nitrogen prediction models and to estimate the distribution of N values in orchards in July. Spatial variance analysis was used to determine optimal sampling strategies.

Results

Leaf nitrogen concentration in summer can be predicted (r²?=?0.9) from the leaf N and B concentration in spring with the sum of K, Ca, and Mg equivalents. Mean field leaf nutrient concentration can be obtained by collecting one pooled sample per management zone composed of 30 trees each of which are at least 30 m apart. Using these methods the percentage of trees with leaf N above the recommended July critical value can be predicted accurately.

Conclusions

Optimized methods for sample collection and models to predict mid-season leaf N from early season samples can be used to improve N management in deciduous tree crops.     

Effect of Particle Shape on Dry Particle Inhalation: Study of Flowability, Aerosolization, and Deposition Properties

The shape effects of dry particles on flowability, aerosolization, and deposition properties in inhalation drug delivery are studied. The properties are compared with similar size range particles of different shapes such as sphere, needle, cube, plate, and pollen. Flowability of the particles is characterized by Carr’s compressibility index and angle of slide (θ) method. The aerosolization and deposition properties of the particles are studied in vitro using an eight-stage Anderson cascade impactor with a Rotahaler®. Pollen-shaped particles are found to exhibit better flowability, higher emitted dose, and higher fine particle fraction than particles of other shapes in similar size range. They showed minimum θ of 35° and maximum emitted dose of 87% and fine particle fraction of 16%. The use of pollen-shaped particles can be a potential improvement in dry particle inhalation.