8S globulin of mungbean [Vigna radiata (L.) Wilczek]: cloning and characterization of its cDNA isoforms, expression in Escherichia coli, purification, and crystallization of the major recombinant 8S isoform

Three isoforms of the cDNA of the major 8S globulin of mungbean, 8Salpha, 8Salpha', and 8Sbeta, were isolated, cloned, and characterized. The cDNA sequences of 8Salpha, 8Salpha', and 8Sbeta had open reading frames of 1362, 1359 or 1362, and 1359 bp, respectively, which code for 454, 453 or 454, and 453 amino acids corresponding to molecular weights of 51 973, 51 627 or 51 758, and 51 779, respectively. Homology in terms of cDNA and amino acid sequences was 91-92% between 8Salpha and 8Salpha', 87% between 8Salpha and 8Sbeta, and 86-88% between 8Salpha' and 8Sbeta. The signal peptide was found to be 1-25, 1-24 or 25, and 1-23 for 8Salpha, 8Salpha', and 8Sbeta, respectively, using the signalP website (Nielsen, H.; Engelbrecht, J.; Brunak, S.; von Heijne, G. Protein Eng. 1997, 10, 1-6). The propeptide was determined to be IVHREN. A single site for glycosylation (N-X-S/T) was observed about 90 amino acids from the C terminus. Homology between mungbean 8S isoforms and other 7-8S proteins ranged from 45 to 68% within members of the legume family and 29 to 34% for crops of different species. The major isoform 8Salpha was expressed in Escherichia coli and purified by successive ammonium sulfate fractionation, hydrophobic interaction, and Mono Q column chromatography. The recombinant 8Salpha, but not the native form, was successfully crystallized producing rhombohedral crystals.     

Measurement of the Lateral Thoracic Reflex Latency in Ponies

Lateral thoracic nerve reflex latencies values were measured in ponies using a simple, non-invasive technique. The reflex was elicited using an external triggering hammer attached to an electrodiagnostic unit. The resulting evoked, compound muscle action potentials were recorded with electrodes, which were placed over the 6th ribs or 11th rib level with the axilla. Two superimposed repeats of 4 signal-averaged sweeps of 50 or 100 milliseconds were recorded and the estimated reflex pathway was measured for each subject in order to calculate the reflex latencies and latency velocities. Mean left and right 6th rib peak latencies were not significantly different from each other ( P = .609), but left 6th rib latencies were shorter than those recorded from the 11th rib ( P < .0001), substantiating the existence of an indirect (central) pathway to the reflex. The calculated left and right 6th rib latency velocities were not significantly different from each other ( P = .58) but left 6th rib latency velocities were different from left 11th rib ( P = .009). The calculated latency velocities were within the broad range for corticospinal tract motor conduction velocities and comparable to magnetic motor evoked latency velocities. The use of lateral thoracic reflex latency measurements to objectively identify the site of spinal cord lesions is discussed.     

Elasmobranch fisheries in the Arabian Seas Region: Characteristics,trade and management

The Arabian Seas Region plays an important role in the global landings and trade of sharks and rays. The United Arab Emirates (UAE) and Yemen, two countries with stark socio‐economic differences, serve as major regional trade hubs for shark and ray products and four countries (Oman, Pakistan, UAE and Yemen) supply nearly 11% of dried fin exports to Hong Kong. Yet, little information is available on the characteristics of this trade and the fisheries contributing to it. Here, we review the fisheries characteristics, trade, utilization and distribution chain of sharks and rays in 15 countries of the Arabian Seas Region based on published and grey literature, landing surveys, field observations and interviews with fishermen and traders. Although regional shark fisheries remain mostly artisanal, reported shark and ray landings represent 28% of the regional total fish production, reaching 56,074 mt in 2012 (7.3% of total world catches), with Iran, Oman, Pakistan and Yemen ranking as the primary catchers. Utilization and distribution patterns are complex, vary between landing sites and countries, and remain unmonitored. Based on widespread over‐exploitation of most teleost fisheries, current exploitation levels for most sharks and rays are potentially unsustainable. The situation is exacerbated by limited research and political will to support policy development, the incomplete nature of fisheries data, as well as insufficient regulations and enforcement. A better understanding of shark and ray fisheries will be key for regulating trade, promoting conservation and developing management initiatives to secure food security, livelihoods and biodiversity conservation in the region.     

Can Batesian mimicry help plants to deter herbivores?

Several authors have suggested that edible plants could avoid herbivory by mimicking olfactory cues of toxic plants. However, very few studies have been carried out to test this hypothesis. The aims of the present study were to identify the volatiles of three clover species and to test whether a species lacking chemical defences, such as red clover, could avoid being grazed by rabbits by mimicking the volatiles of the cyanogenic white clover. Two main volatiles were identified in all three clover species, and a further two volatiles were present in white clover only. Rabbits presented with a choice between white clover, red clover and red clover sprayed with white clover extract ate significantly more red clover than white or white-flavoured red clover. The results suggest that the volatiles of toxic plants could be used and exploited as a source of natural, safe and effective repellents to control the impact of pest herbivores on plants.     

Evaluation of electroacupuncture treatment for thoracolumbar intervertebral disk disease in dogs

OBJECTIVE: To evaluate use of electroacupuncture combined with standard Western medical treatment versus Western medical treatment alone for treatment of thoracolumbar intervertebral disk disease in dogs. DESIGN: Prospective controlled study. ANIMALS: 50 dogs with signs of thoracolumbar intervertebral disk disease. PROCEDURES: Dogs were randomly allocated to 1 of 2 treatment groups and classified as having grade 1 to 5 neurologic dysfunction. Dogs in group 1 received electroacupuncture stimulation combined with standard Western medical treatment; those in group 2 received only standard Western medical treatment. A numeric score for neurologic function was evaluated at 4 time points to evaluate effects of treatments. RESULTS: Time (mean +/- SD) to recover ambulation in dogs with grade 3 and 4 dysfunction in group 1 (10.10 +/- 6.49 days) was significantly lower than in group 2 (20.83 +/- 11.99 days). Success (able to walk without assistance) rate for dogs with grade 3 and 4 dysfunction in group 1 (10/10 dogs) was significantly higher than that of similarly affected dogs in group 2 (6/9 dogs). Dogs without deep pain perception (grade 5 dysfunction) had a success (recovery of pain sensation) rate of 3 of 6 and 1 of 8 in groups 1 and 2, respectively, but the difference was not significant. Overall success rate (all dysfunction grades) for group 1 (23/26; 88.5%) was significantly higher than for group 2 (14/24; 58.3%). CONCLUSIONS AND CLINICAL RELEVANCE: Electroacupuncture combined with standard Western medical treatment was effective and resulted in shorter time to recover ambulation and deep pain perception than did use of Western treatment alone in dogs with signs of thoracolumbar intervertebral disk disease.     

Protective Effect of <Emphasis Type="Italic">Juglans regia</Emphasis> L. Walnut Extract Against Oxidative DNA Damage

Walnuts (Juglans regia L.) are relevant components of the Mediterranean diet providing important macronutrients, micronutrients and other bioactive constituents including unsaturated fatty acids, proteins, fiber, vitamins, minerals, phytosterols and polyphenols. Although the walnut beneficial effects in human health are widely recognized by a lot of epidemiologic studies very little is known regarding its effect on damaged DNA. The aim of the present study was to investigate the effect of Juglans regia L. ethanolic extract from kernel on the induction of DNA strand breaks by thiol/Fe³⁺/O₂ mixed function oxidase, tert-butyl hydroperoxide or UVC radiations in acellular and cellular models. Plasmid DNA cleavage and fast Halo assay were used to monitor oxidative damage to DNA. Both approaches showed protection of oxidatively injured DNA. These results agree with a lot of scientific proofs which recommend walnut as dietary adjunct in health promotion and prevention as well as in treatment of lifestyle-related oxidative diseases.     

Demographic network and multi-season occupancy modeling of Rana sylvatica reveal spatial and temporal patterns of population connectivity and persistence

Many populations are spatially structured with frequent extinction–colonization events. A clear understanding of these processes is necessary for making informed and effective management decisions. Due to the spatially and temporally dynamic nature of many systems, population connectivity and local extinction–colonization processes can be difficult to assess, but graph theoretic and occupancy modeling approaches are increasingly being utilized to answer such vital ecological questions. In our study, we used 6 years of egg mass counts from 34 ponds for Rana sylvatica to parameterize spatially explicit demographic network models. Our models revealed that the studied populations have spatial structure with strong source–sink dynamics. We also assessed the colonization and persistence probability of each pond using multi-season occupancy modeling. We observed extreme fluctuation in reproductive effort among years, resulting in variable levels of connectivity across the landscape. Pond colonization and persistence were most influenced by local population dynamics, but colonization was also affected by precipitation. Our demographic network model had moderate ability to predict reproductive effort, but accuracy was hindered by variation in annual precipitation. Source populations had higher colonization and persistence rates as well as a greater proportion of ravine habitat within 1,000 m than sink populations. By linking a spatially explicit connectivity model with a temporal occupancy/persistence model, we provide a framework for interpreting patterns of occupancy and dispersal that can serve as an initial guide for future habitat management and restoration.     

Genetic analysis and genotype × environment (G × E) for grey leaf spot disease resistance in elite African maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) germplasm

Maize grey leaf spot (GLS) disease remains an important foliar disease in sub-Saharan Africa accounting for more than 25% yield losses in maize. Information on inheritance of GLS resistance of germplasm adapted to African environments is required in new sources being identified. Therefore, hybrids generated from a 10 × 10 half-diallel mating of tropical advanced maize inbred lines were evaluated in six environments to determine combining ability, genotype × environment interaction (G × E) and the impact of GLS disease on grain yield. General combining ability effects were highly significant and accounted for 72 and 68% of the variation for GLS resistance and grain yield, respectively. Significant specific combining ability effects associated with reduced disease levels were observed in some hybrids when one parent was resistant, and these may be exploited in developing single cross maize hybrids. Regression analysis showed a 260–320 kg ha^?1 decrease in maize grain yield per each increase in GLS disease severity score, and significant associations (r = ?0.31 to ?0.60) were observed between grain yield and GLS severity scores. This showed the potential of GLS disease to reduce yield in susceptible varieties grown under favourable disease conditions, without control measures. Genotype and genotype × environment biplots and correlation analysis indicated that the significant G × E observed was not due to changes in hybrid ranking, implying absence of a significant crossover interaction. Therefore, predominance of additive gene effects imply that breeding progress for GLS disease resistance would be made through selection and this could be achieved at a few hot-spot sites, such as Baynesfield and Cedara locations in South Africa, and still deploy the resistant germplasm to other environments in which they are adapted.     

Optimizing nitrogen use efficiency in wheat and potatoes: interactions between genotypes and agronomic practices

One approach to decrease the environmental impact of crop production and reduce costs is to optimize agronomic practices and genotypes so that nutrients are used more efficiently. In this study the effects of agronomic practices (rotations, crop protection, fertilization) on yields, nitrogen use efficiency (NUE) and associated parameters were studied in an experiment using two winter wheat genotypes (Cordiale and Scaro) in one season and two potato genotypes (Sarpo Mira and Sante) in two seasons. The wheat showed no varietal differences in yield and NUE; instead the fertilization regime was the main factor affecting yield and NUE with higher values observed when conventional fertilization was used. The exception was for wheat grown after three years grass/clover ley when there was no added yield benefit from conventional fertilization of the organically bred variety (Scaro). This demonstrates the potential for N fixing crops to provide sufficient N to high yielding cereals if grown for long enough prior to planting. The greatest gains in NUE were achieved by combining an N efficient genotype with conventional crop management in an organic rotation. Fertilization and genotypic variation were the main factors affecting potato tuber yield and NUE, with the late maturing Sarpo Mira displaying elevated yields and NUE compared with the early maturing Sante. The use of organic fertility sources resulted in lower NUE, but N release from organic sources may increase NUE of future crops. This highlights the need for long-term nutrient balance and modelling studies to assess NUE at the crop rotation scale.     

Protective activity against oocyst shedding in cats vaccinated with crude rhoptry proteins of the Toxoplasma gondii by the intranasal route

This study evaluated a vaccine made from crude rhoptry proteins of Toxoplasma gondii with Quil-A, which was administered to cats by the intranasal route. Eleven short-hair domestic cats were divided into four groups: G1 (n=3) received three doses (200 microg/dose) of the rhoptry vaccine with Quil-A (20 microg); G2 (n=3) received PBS with Quil-A (20 microg); G3 (n=3) and G4 (n=2) received only PBS. Treatments were administered at days 0, 21, and 42 by the intranasal route. Challenge was done to G1, G2, and G3 animals with 600 cysts of the VEG strain on day 51 (challenge day); G4 animals were unchallenged. The anti-T. gondii IgG and IgA antibody levels from sera were measured by indirect enzyme-linked immunosorbent assay (ELISA). At challenge, two animals from G1 revealed antibody levels for both IgG and IgA; oocysts were not detected in feces of these two cats. There were no differences in hematological values between groups throughout the experiment (p>0.10). Preventable fractions were 67% in G1 and 0% in G2 and G3. Comparatively, G1 animals shed 89.3% and 90.8% less oocysts than G3 and G4, respectively. Two out of three cats were protected against T. gondii oocyst shedding when the rhoptry vaccine was administered by the intranasal route. This is the first study using crude rhoptry proteins as vaccine by the intranasal route in cats to evaluate protection against oocysts shedding.