Characterisation of digestive protease in the rose sawfly,Arge rosae Linnaeus (Hymenoptera: Argidae)

Insect midgut proteases are excellent targets for insecticidal agents such as protease inhibitors. These inhibitors are used for producing transgenic plants, resistant to pests. For achieving this goal, it is necessary to find the nature of specific proteases and their properties for adopting possible pest management procedure. Therefore, characterisation of the enzymes in the gut of the rose sawfly, Arge rosae (Hymenoptera: Argidae), responsible for proteolysis, was performed using a range of synthetic substrates and specific inhibitors. The optimum conditions for general proteases and trypsin were achieved at pH 10. The highest activity for general proteases was obtained at a temperature of 45°C. The use of specific inhibitors and SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) provided evidence to suggest that most of the proteases belonged to the serine group because of high inhibitory effect of phenyl methane sulfonyl fluoride on total proteolytic activity. Also, inhibition assays and zymogram analysis showed that metalloproteases are present in A. rosae digestive system. These results indicated that A. rosae larvae mainly used serine proteases for protein digestion, with chymotrypsin as the dominant form. The kinetic parameters of trypsin-like proteases using N-benzoyl-dl-arg-p-nitroanilide as substrate indicated that the K _m and V _max values of trypsin in the gut of the fifth instar larvae were 730 ± 17.3 μM and 456 ± 13.85 nmol min^?1 mg^?1 protein, respectively.     

Rubella virus E2 signal peptide is required for perinuclear localization of capsid protein and virus assembly

The rubella virus (RV) structural proteins capsid, E2, and E1 are synthesized as a polyprotein precursor. The signal peptide that initiates translocation of E2 into the lumen of the endoplasmic reticulum remains attached to the carboxy terminus of the capsid protein after cleavage by signal peptidase. Among togaviruses, this feature is unique to RV. The E2 signal peptide has previously been shown to function as a membrane anchor for the capsid protein. In the present study, we demonstrate that this domain is required for RV glycoprotein-dependent localization of the capsid protein to the juxtanuclear region and subsequent virus assembly at the Golgi complex.     

Effects of temperature and photoperiod on postponing bermudagrass (Cynodon dactylon [L.] Pers.) turf dormancy

Growth chamber and field experiments were carried out to determine the effects of extended photoperiod under low and freezing temperatures on bermudagrass turf dormancy at Bajgah, in the southern part of Iran. The experiment in the growth chamber was conducted with four temperature regimes (15, 7.5, 0 and -7.5°C) and three light durations (8, 12 and 16h) in a completely randomized design with four replications. The field study was conducted in two consecutive years (2008-2009) with three light durations (8, 12 and 16h) in months with natural short day length and arranged in a randomized complete blocks design with three replications. Results in both experiments showed that decreasing temperature and photoperiod decreased verdure fresh and dry weight, shoot height, tiller density, leaf area and chlorophyll and relative water contents (RWC). However, rooting depth and fresh weight of roots increased in the growth chamber. Decreasing the temperature and light duration increased electrolyte leakage and proline content. Reducing sugars increased with decreasing temperature and declined with lowering light duration in both shoots and roots. Starch content of both shoots and roots showed an adverse trend compared to reducing sugars; starch content increased in both shoots and roots in all treatments by shortening the photoperiod. Practically, the problem of bermudagrass turf's dormancy could be solved via increasing the photoperiod in months with short day lengths. This treatment would be efficient and useful for turfgrass managers to apply in landscapes and stadiums.     

Oral poliovirus vaccine in management of recurrent aphthous stomatitis

Oral poliovirus vaccine (OPV) is reported to be effective in treatment of recurrent herpes simplex (RHS). According to our observation during recent years, OPV was not only effective in management of RHS but also in some patients with concomitant recurrent aphthous stomatitis (RAS) reducing its severity and frequency. The purpose of this study was to evaluate the efficacy of OPV in the management of RAS. In a longitudinal, case--control study 48 patients with RAS were recruited. Twenty patients received OPV and 28 patients received placebo. OPV was administered in a dose of 4 drops at monthly intervals for 3 months to the study group while the control group received placebo. The results were registered in 3 months after the last dose. Eight cases (40%) in the OPV group showed significant reduction in the duration of the ulcers, while no change was seen in the control group (P = 0.048). The frequency of recurrence of RAS was reduced in 13 cases (65%) in the OPV group, and in 6 cases (21.4%) of the placebo group (P = 0.006). The severity of attacks was reduced in 12 cases (60%) in the OPV group and in 4 cases (14.3%) in the placebo group (P = 0.008). In conclusion OPV appeared to be effective in the management of RAS.     

Ovocalyxin-36 Is a Pattern Recognition Protein in Chicken Eggshell Membranes

The avian eggshell membranes are essential elements in the fabrication of the calcified shell as a defense against bacterial penetration. Ovocalyxin-36 (OCX-36) is an abundant avian eggshell membrane protein, which shares protein sequence homology to bactericidal permeability-increasing protein (BPI), lipopolysaccharide-binding protein (LBP) and palate, lung and nasal epithelium clone (PLUNC) proteins. We have developed an efficient method to extract OCX-36 from chicken eggshell membranes for purification with cation and anion exchange chromatographies. Purified OCX-36 protein exhibited lipopolysaccharide (LPS) binding activity and bound lipopolysaccharide (LPS) from Escherichia coli O111:B4 in a dose-dependent manner. OCX-36 showed inhibitory activity against growth of Staphylococcus aureus ATCC 6538. OCX-36 single nucleotide polymorphisms (SNPs) were verified at cDNA 211 position and the corresponding proteins proline-71 (Pro-71) or serine-71 (Ser-71) were purified from eggs collected from genotyped hens. A significant difference between Pro-71 and Ser-71 OCX-36 for S. aureus lipoteichoic acid (LTA) binding activity was detected. The current study is a starting point to understand the innate immune role that OCX-36 may play in protection against bacterial invasion of both embryonated eggs (relevant to avian reproductive success) and unfertilized table eggs (relevant to food safety).     

Can audio–visual or visual stimuli from a prospective mate stimulate a reproductive neuroendocrine response in sheep?

Stimuli from a prospective mate increase the secretion of luteinising hormone (LH) in sheep. This 'male effect' in ewes and 'female effect' effect in rams is predominantly mediated by olfactory signals, though it is thought that non-olfactory signals play synergistic or substitutive roles. In this study, we tested whether exposure to visual or audio-visual stimuli from a prospective mate would stimulate an increase in LH secretion in ewes (Experiment 1) and rams (Experiment 2). In Experiment 1, groups of eight Merino ewes were exposed to one of three stimuli midway through a frequent blood-sampling regimen: full ram contact, still images of rams, a video of ewes and rams mating. Control ewes (n = 8) were completely isolated from rams. Exposure to still images of rams appeared to stimulate an increase in mean LH concentrations (P < 0.05) and tended to increase LH pulse frequency (P < 0.1), but the response was significantly smaller than that observed in ewes exposed to rams (P < 0.01). Audio-visual stimuli had no effect on any parameters of LH secretion (P > 0.1). In Experiment 2, Merino rams were allocated to either an Exposure (n = 7) or a Control (n = 7) group. Exposure rams underwent two exposure periods midway through a frequent blood-sampling regimen; exposure to still images of ewes and audio recorded during mating of ewes and rams (audio-visual exposure); exposure to oestrous ewes (ewe exposure). Control rams were sampled at the same frequency but remained isolated from ewe stimuli. Exposure of rams to the audio-visual stimuli did not affect any parameters of LH secretion (P > 0.1). In contrast, exposure to oestrous ewes increased LH pulse frequency (P < 0.05) and advanced the onset of the next LH pulse (P < 0.05). In conclusion, visual signals appear to be involved in eliciting the neuroendocrine response of ewes to rams and are of greater importance to this phenomenon in ewes (male effect) than rams (female effect). However, overall the visual and audio-visual signals used in this study were far less effective than stimulus animals, suggesting that these stimuli are less important than olfactory signals, or a combination of olfactory and audio-visual signals.