Conserved water-mediated H-bonding dynamics of catalytic Asn 175 in plant thiol protease

The role of invariant water molecules in the activity of plant cysteine protease is ubiquitous in nature. On analysing the 11 different Protein DataBank (PDB) structures of plant thiol proteases, the two invariant water molecules W I and W2 (W220 and W222 in the template 1PPN structure) were observed to form H-bonds with the Ob atom of Asn 175. Extensive energy minimization and molecular dynamics simulation studies up to 2 ns on all the PDB and solvated structures clearly revealed the involvement of the H-bonding association of the two water molecules in fixing the orientation of the asparagine residue of the catalytic triad. From this study, it is suggested that H-bonding of the water molecule at the W1 invariant site better stabilizes the Asn residue at the active site of the catalytic triad.

Perennial grassland dynamics on fertile plains: Is coexistence mediated by disturbance?

The response of grasslands to disturbance varies with the nature of the disturbance and the productivity of the landscape. In highly productive grasslands, competitive exclusion often results in decreased species richness and grazing may allow more species to coexist. Once widespread, grasslands dominated by Dichanthium sericeum (Queensland bluegrass) and Astrebla spp. (Mitchell grass) occur on fertile plains but have been reduced in extent by cultivation. We tested the effects of exclusion of livestock grazing on these grasslands by comparing the floristic composition of sites in a nature reserve with an adjacent stock reserve. In addition, sites that had been cultivated within the nature reserve were compared with those where grazing but no cultivation had occurred. To partition the effects of temporal variation from spatial variation we sampled sites in three different years (1998, 2002 and 2004). Some 194 taxa were recorded at the nature reserve and surrounding stock routes. Sampling time, the occurrence of past cultivation and livestock grazing all influenced species composition. Species richness varied greatly between sampling periods relating to highly variable rainfall and water availability on heavy clay soils. Native species richness was significantly lower at previously cultivated sites (13-22 years after cultivation), but was not significantly influenced by grazing exclusion. After 8 years it appears that reintroducing disturbance in the form of livestock grazing is not necessary to maintain plant species richness in the reserve. The highly variable climate (e.g. droughts) probably plays an important role in the coexistence of species by negating competitive exclusion and allowing interstitial species to persist.

Multiple-quantum artifacts in single-quantum two-dimensional correlated NMR spectra of strongly coupled spins

Artifacts in the form of cross peaks have been observed along two- and three-quantum diagonals in single-quantum two-dimensional correlated (COSY) spectra of several peptides and oligonucleotides. These have been identified as due to the presence of a non-equilibrium state of kind I (a state describable by populations which differ from equilibrium) of strongly coupled spins carried over from one experiment to the next in the COSY algorithm.

Personality and social psychological studies on computer-mediated communication in school settings

The present study examined how personality and social psychological factors affect third and fourth graders' computer-mediated communication. Personality was analysed in terms of the following strategies: optimism, pessimism and defensive pessimism. Students worked either individually or in dyads which were paired homogeneously or heterogeneously according to the strategies. Moreover, the present study compared horizontal and vertical interaction. The study also examined the role that popularity plays, and students were divided into groups based on their popularity level. The results show that an optimistic strategy is useful. Optimism was found to be related to the active production and processing of ideas. Although previous research has identified drawbacks to pessimism in achievement settings, this study shows that the pessimistic strategy is not as debilitating a strategy as is usually assumed. Pessimistic students were able to process their ideas. However, defensive pessimists were somewhat cautious in introducing or changing ideas. Heterogeneous dyads were not beneficial configurations with respect to producing, introducing, or changing ideas. Moreover, many differences were found to exist between the horizontal and vertical interaction; specifically, the students expressed more opinions and feelings when teachers took no part in the discussions. Strong emotions were observed especially in the horizontal interaction. Further, group working skills were found to be more important for boys than for girls, while rejected students were not at a disadvantage compared to popular ones. Schools can encourage emotional and social learning. The present study shows that students can use computers to express their feelings. In addition, students who are unpopular in non-computer contexts or students who use pessimism can benefit from computers. Participation in computer discussions can give unpopular children a chance to develop confidence when relating to peers.

Management of plant invasions mediated by frugivore interactions

1. Some of the most damaging invasive plants are dispersed by frugivores and this is an area of emerging importance in weed management. It highlights the need for practical information on how frugivores affect weed population dynamics and spread, how frugivore populations are affected by weeds and what management recommendations are available. 2. Fruit traits influence frugivore choice. Fruit size, the presence of an inedible peel, defensive chemistry, crop size and phenology may all be useful traits for consideration in screening and eradication programmes. By considering the effect of these traits on the probability, quality and quantity of seed dispersal, it may be possible to rank invasive species by their desirability to frugivores. Fruit traits can also be manipulated with biocontrol agents. 3. Functional groups of frugivores can be assembled according to broad species groupings, and further refined according to size, gape size, pre- and post-ingestion processing techniques and movement patterns, to predict dispersal and establishment patterns for plant introductions. 4. Landscape fragmentation can increase frugivore dispersal of invasives, as many invasive plants and dispersers readily use disturbed matrix environments and fragment edges. Dispersal to particular landscape features, such as perches and edges, can be manipulated to function as seed sinks if control measures are concentrated in these areas. 5. Where invasive plants comprise part of the diet of native frugivores, there may be a conservation conflict between control of the invasive and maintaining populations of the native frugivore, especially where other threats such as habitat destruction have reduced populations of native fruit species. 6. Synthesis and applications. Development of functional groups of frugivore-dispersed invasive plants and dispersers will enable us to develop predictions for novel dispersal interactions at both population and community scales. Increasingly sophisticated mechanistic seed dispersal models combined with spatially explicit simulations show much promise for providing weed managers with the information they need to develop strategies for surveying, eradicating and managing plant invasions. Possible conservation conflicts mean that understanding the nature of the invasive plant-frugivore interaction is essential for determining appropriate management.

Direct estimation of an element of order matrix from H-1 NMR spectra of strongly dipolar coupled spins

NMR spectra of molecules oriented in thermotropic liquid crystalline media provide information on the molecular structure and order. The spins are generally strongly dipolar coupled and the spectral analyse require the tedious and time consuming numerical iterative calculations. The present study demonstrates the application of multiple quantum spin state selective detection of single quantum transitions for mimicking the homonuclear decoupling and the direct estimation of an element of ordering matrix. This information is utilized to estimate the nearly accurate starting dipolar couplings for iterative calculations. The studies on the spectra of strongly dipolar coupled five and six interacting spin systems are reported.

Microemulsion-mediated synthesis of cobalt (pure fcc and hexagonal phases) and cobalt-nickel alloy nanoparticles

By choosing appropriate microemulsion systems, hexagonal cobalt (Co) and cobalt-nickel (1:1) alloy nanoparticles have been obtained with cetyltrimethylammonium bromide as a cationic surfactant at 500 degrees C. This method thus stabilizes the hcp cobalt even at sizes (<10 nm) at which normally fcc cobalt is predicted to be stable. On annealing the hcp cobalt nanoparticles in H-2 at 700 degrees C we could transform them to fcc cobalt nanoparticles. Microscopy studies show the formation of spherical nanoparticles of hexagonal and cubic forms of cobalt and Co-Ni (1:1) alloy nanoparticles with the average size of 4, 8 and 20 nm, respectively. Electrochemical studies show that the catalytic property towards oxygen evolution is dependent on the applied voltage. At low voltage (less than 0.65 V) the Co (hexagonal) nanoparticles are superior to the alloy (Co-Ni) nanoparticles while above this voltage the alloy nanoparticles are more efficient catalysts. The nanoparticles of cobalt (hcp and fcc) and alloy (Co-Ni) nanoparticles show ferromagnetism. The saturation magnetization of Co-Ni nanoparticles is reduced compared to the bulk possibly due to surface oxidation.

Borax Mediated Layer-by-Layer Self-Assembly of Neutral Poly(vinyl alcohol) and Chitosan

We report a multilayer film of poly(vinyl alcohol) (PVA)-borate complex and chitosan by using a layer-by-layer approach. PVA is an uncharged polymer, but hydroxyl functional groups of PVA can be crosslinked by using borax as a cross-linking agent. As a result electrostatic charges and intra- and interchain cross-links are introduced in the PVA chain and provide physically cross-linked networks. The PVA-borate was then deposited on a flat Substrate as well as on colloidal particles with chitosan as an oppositely charged polyelectrolyte. Quartz crystal microbalance. scanning electron microscopy, and atomic force microscopy were used to follow the growth of thin film oil flat substrate. Analogous experiments were performed on melamine formaldehyde colloidal particles (3-3.5 mu m) to quantify the process for the preparation of hollow rnicrocapsules. Removal of the core in 0.1 N HCI results in hollow microcapsules. Characterization of microcapsules by transmission electron microscopy revealed formation of stable microcapsules. Further, self-assembly of PVA-borate/chitosan was loaded with the anticancer drug doxorubicin, and release rates were determined at different pH Values to highlight the drug delivery potential of this system.

Hierarchical Bayesian modelling of plant pest invasions with human-mediated dispersal

Hierarchical Bayesian models can assimilate surveillance and ecological information to estimate both invasion extent and model parameters for invading plant pests spread by people. A reliability analysis framework that can accommodate multiple dispersal modes is developed to estimate human-mediated dispersal parameters for an invasive species. Uncertainty in the observation process is modelled by accounting for local natural spread and population growth within spatial units. Broad scale incursion dynamics are based on a mechanistic gravity model with a Weibull distribution modification to incorporate a local pest build-up phase. The model uses Markov chain Monte Carlo simulations to infer the probability of colonisation times for discrete spatial units and to estimate connectivity parameters between these units. The hierarchical Bayesian model with observational and ecological components is applied to a surveillance dataset for a spiralling whitefly (Aleurodicus dispersus) invasion in Queensland, Australia. The model structure provides a useful application that draws on surveillance data and ecological knowledge that can be used to manage the risk of pest movement.

Gene expression evidence for off-target effects caused by RNA interference-mediated gene silencing of Ubiquitin-63E in the cattle tick Rhipicephalus microplus.

Knowledge of cattle tick (Rhipicephalus (Boophilus) microplus; Acari: Ixodidae) molecular and cellular pathways has been hampered by the lack of an annotated genome. In addition, most of the tick expressed sequence tags (ESTs) available to date consist of similar to 50% unassigned sequences without predicted functions. The most common approach to address this has been the application of RNA interference (RNAi) methods to investigate genes and their pathways. This approach has been widely adopted in tick research despite minimal knowledge of the tick RNAi pathway and double-stranded RNA (dsRNA) uptake mechanisms. A strong knockdown phenotype of adult female ticks had previously been observed using a 594 bp dsRNA targeting the cattle tick homologue for the Drosophila Ubiquitin-63E gene leading to nil or deformed eggs. A NimbleGen cattle tick custom microarray based on the BmiGI.V2 database of R. microplus ESTs was used to evaluate the expression of mRNAs harvested from ticks treated with the tick Ubiquitin-63E 594 bp dsRNA compared with controls. A total of 144 ESTs including TC6372 (Ubiquitin-63E) were down-regulated with 136 ESTs up-regulated following treatment. The results obtained substantiated the knockdown phenotype with ESTs identified as being associated with ubiquitin proteolysis as well as oogenesis, embryogenesis, fatty acid synthesis and stress responses. A bioinformatics analysis was undertaken to predict off-target effects (OTE) resulting from the in silico dicing of the 594 bp Ubiquitin-63E dsRNA which identified 10 down-regulated ESTs (including TC6372) within the list of differentially expressed probes on the microarrays. Subsequent knockdown experiments utilising 196 and 109 bp dsRNAs, and a cocktail of short hairpin RNAs (shRNA) targeting Ubiquitin-63E, demonstrated similar phenotypes for the dsRNAs but nil effect following shRNA treatment. Quantitative reverse transcriptase PCR analysis confirmed differential expression of TC6372 and selected ESTs. Our study demonstrated the minimisation of predicted OTEs in the shorter dsRNA treatments (similar to 100-200 bp) and the usefulness of microarrays to study knockdown phenotypes.

Ionophore-mediated transmembrane movement of divalent cations in small unilamellar liposomes: An evaluation of the chlortetracycline fluorescence technique and correlations with black lipid membrane studies

Conceptual advances in the field of membrane transport have, in the main, utilized artificial membranes, both planar and vesicular. Systems of biological interest,viz., cells and organelles, resemble vesicles in size and geometry. Methods are, therefore, required to extend the results obtained with planar membranes to liposome systems. In this report we present an analysis of a fluorescence technique, using the divalent cation probe chlortetracycline, in small, unilamellar vesicles, for the study of divalent cation fluxes. An ion carrier (X537 A) and a pore former (alamethicin) have been studied. The rate of rise of fluorescence signal and the transmembrane ion gradient have been related to transmembrane current and potential, respectively. A second power dependence of ion conduction-including the electrically silent portion thereof — on X537 A concentration, has been observed. An exponential dependence of ldquocurrentrdquo on ldquotransmembrane potentialrdquo in the case of alamethicin is also confirmed. Possible errors in the technique are discussed.

Inhibition of cortisol-mediated induction of hepatic tryptophan-2,3-dioxygenase by norepinephrine

Norepinephrine inhibits cortisol-mediated induction of hepatic tryptophan pyrrolase in rats. During cold exposure the stabilization of this enzyme appears to occur by an interaction of corticoids and norepinephrine on the induction process.

Silica vesicles as nanocarriers and adjuvants for generating both antibody and T-cell mediated immune resposes to Bovine Viral Diarrhoea Virus E2 protein

Bovine Viral Diarrhoea Virus (BVDV) is widely distributed in cattle industries and causes significant economic losses worldwide annually. A limiting factor in the development of subunit vaccines for BVDV is the need to elicit both antibody and T-cell-mediated immunity as well as addressing the toxicity of adjuvants. In this study, we have prepared novel silica vesicles (SV) as the new generation antigen carriers and adjuvants. With small particle size of 50 nm, thin wall (similar to 6 nm), large cavity (similar to 40 nm) and large entrance size (5.9 nm for SV-100 and 16 nm for SV-140), the SV showed high loading capacity (similar to 250 mu g/mg) and controlled release of codon-optimised E2 (oE2) protein, a major immunogenic determinant of BVDV. The in vivo functionality of the system was validated in mice immunisation trials comparing oE2 plus Quil A (50 mu g of oE2 plus 10 mu g of Quil A, a conventional adjuvant) to the oE2/SV-140 (50 mu g of oE2 adsorbed to 250 mu g of SV-140) or oE2/SV-140 together with 10 mu g of Quil A. Compared to the oE2 plus Quil A, which generated BVDV specific antibody responses at a titre of 10(4), the oE2/SV-140 group induced a 10 times higher antibody response. In addition, the cell-mediated response, which is essential to recognise and eliminate the invading pathogens, was also found to be higher [1954-2628 spot forming units (SFU)/million cells] in mice immunised with oE2/SV-140 in comparison to oE2 plus Quil A (512-1369 SFU/million cells). Our study has demonstrated that SV can be used as the next-generation nanocarriers and adjuvants for enhanced veterinary vaccine delivery. (C) 2014 Elsevier Ltd. All rights reserved.