Volume 70, Issue 12

Recurrent outbreaks of Newcastle disease have questioned the usage of existing vaccines that whether they are still adequate to protect clinical diseases and inhibit virus transmission in poultry. Advancement in molecular biology has led to the production of recombinant vaccines in recent years, which can be a more useful strategy to control infections of Newcastle disease virus (NDV). Studies indicate that the pathogenic nature of NDV is mediated by its membrane associated fusion (F) protein. Here we report the cloning of the full-length F gene-pET30a and its expression in Escherichia coli BL21 DE3 cells through isopropyl β-D-1-thiogalactopyranoside induction. Transferring the protein on nitrocellulose membrane in Western blotting confirmed its specificity with histidine-tagged antibody reaction at the proper size of 67 kDa. Protein purification with nickel charged sepharose column affinity chromatography resulted in a single band of 67 kDa purified His-tag F protein on SDS-PAGE. Analysis of its immunogenicity through bioinformatics tools revealed that more than 70% of its sequence is antigenically active comprising 24 linear immunogenic peptides predicted by the Linear epitope prediction tool and 9 immunogenic peptides predicted by ElliPro. This is a key achievement of the study, which may lead towards recombinant vaccine production in future. In conclusion, our findings suggest that rather than employing live viral vaccines, using a purified immunogenic recombinant F protein as a vaccine or cloning the same gene in a suitable plant vector for production of edible vaccine will provide better protection against the NDV into chicken.

A potential screening programme for fibrinolytic filamentous fungi in the Egyptian environment was done for the first time. The proteolytic activity was positive in 38.5% of isolates, whilst only thirty-four of them were able to produce fibrinolytic enzymes. The two most potent isolates were identified as Aspergillus brasiliensis AUMC 9735 and Aspergillus flavus AUMC 9736. Under submerged culture conditions, the two strains were able to excrete fibrinolytic enzymes, reaching a maximum at 5 and 9 days, respectively. Maximal enzyme productivity by both strains was achieved by lactose and sucrose, respectively. All tested nitrogen sources were stimulatory for enzyme production by both strains, except for ammonium acetate in the case of A. brasiliensis AUMC 9735. Purification of A. brasiliensis AUMC 9735 enzyme increased its specific activity to 83.5-fold with a recovery of 9.1% and molecular weight of 40 kDa. Maximal activity was recorded at pH 8 (stability at pH 6-11). The purified enzyme showed higher thermostability than most fibrinolytic enzymes of filamentous fungi with the midpoint temperature (T m ) value at 60.4°C and the half-life times (t 1/2 ) at 50-80°C being 672.1-6.5 min. Interestingly, the enzyme exhibited a good storage and solvent stability at 4°C for 60 days. Enzyme activity was totally lost in the presence of ethylenediaminetetraacetic acid and was restored after addition of Mg 2+ cations suggesting that the enzyme is a metalloprotease and Mg 2+ acts as a cofactor. This work reveals the potential of A. brasiliensis and many other fungal strains as an unconventional and unexplored production alternative to already known thrombolytic agents. The value of Egyptian environment as a reservoir of filamentous fungal species with promising biotechnological value could also be highlighted.

The effect of copper oxide nanoparticles (CuONPs) at the biochemical, anatomical and molecular level was investigated in cucumber (Cucumis sativus L.) seedlings. The seedlings were grown in semi-solid half strength Murashige and Skoog medium supplemented with 0, 50, 100, 200, 400 and 500 mg/L of CuONPs for fifteen days under controlled growth chamber conditions. The results showed that exposure to all concentrations of CuONPs resulted in significant reduction in shoot and root growth and biomass. A concentration-dependant increase in reactive oxygen species (ROS) generation, malondialdehyde production, lignin content and decline in mitochondrial membrane potential were observed. Cross-sections of stem showed anatomical changes, viz. an increase in xylogenesis in CuONPs exposed plants. Significant modulation in the expression of genes related to oxidative stress and lignin biosynthesis, i.e. catalase, ascorbate peroxidase, phenylalanine ammonia lyase, cinnamate 4-hydroxylase, anionic and cationic peroxidases were observed in shoots and roots as a result of CuONPs exposure. Taken together, exposure to CuONPs has resulted in excess ROS generation, lignification and growth suppression in Cucumis sativus seedlings.

Changes in expression of glutamine synthetase (GS) have effect on plant nitrogen metabolism. In order to improve nitrogen use efficiency, several attempts at over-expressing GS II genes in plants have been previously undertaken, however few GSI and III genes are found such application. In this study, two GS I genes were cloned from bacterial strains and were transformed into Arabidopsis thaliana. Expression of the genes was confirmed under both mRNA and protein levels. Phenotypic studies revealed that all transgenic Arabidopsis lines showed enhanced fresh weight (12%) and dry weight (13%) compared with the wild-type plants at two concentrations of nitrate supplies. Further biochemical characterization confirmed that the transgenic lines had higher total nitrogen content (increased by 5-8%), soluble protein concentration (increased by 7-11%), total amino acid content (increased by 4-8%), leaf GS activity (enhanced by 8-14%) and free NO - 3 concentration (increased by 8-16%) compared to wild type Arabidopsis. This work firstly reported that over-expression of bacterial GS I genes in Arabidopsis resulted in improvement of growth phenotype and nitrogen use efficiency (NUE) of plant. The bacterial GS genes could be potentially useful in engineering transgenic plants of high NUE.

Many lakes in mountain regions have been used for hydropower generation since the 1950s. It has been estimated that as many as 79% of the rivers in the Alps have been affected by the presence of hydropower plants. In this context, the shutting down of hydropower plants on a group of Alpine lakes represented a good opportunity to study the ecological impact on them. We selected nine lakes that had been affected and nine that had not, and analysed the differences in environment, littoral diatoms and zoobenthos, phytoplankton, zooplankton, and fish. Results showed that benthic biota -diatoms and zoobenthos- were the most affected by water-level drawdown during winter months. Even six years after the end of hydroelectric operations, diatom species richness and diversity were lower in impacted lakes. Assemblage structure was different for both diatoms and zoobenthos. Phytoplankton and zooplankton were similar in impacted and unaffected lakes in terms of both species richness (and diversity) and assemblage structure. The degree of impact on fish was unclear because illegal stocking of lakes with allochthonous fish species had taken place. This study showed that compared to limnetic biota, littoral communities were the most affected by the decrease in water volume every winter. Six years after the end of hydroelectric operations, diatoms, and to lesser extent zoobenthos, were still different compared to those in natural (unaffected) lakes. Planktic communities seem to be either more resistant to the disturbances, or else able to recover more quickly to their former condition.

In the bodies of seed plants, lateral organs of the shoot, for example cotyledons, leaves, and floral organs, are determinate and exhibit localized planer growth resulting in breaking of radial symmetry and asymmetric development. Localized planer growth in the leaf generates the leaf blade, the principle site of photosynthesis in most plants. Development of flat and expanded lamina is an important physiological process. In these processes, a few kind of meristems cooperate with regulating leaf development. In this work, with transfer sense ASL11, sense ASL38 and antisense ASL15 under 35S promoter to to Arabidopsis (Col-0), we found a series of radial symmetric needle-like rosette leaves. By analyzing the anatomical and the epidermal cell features, our data suggests that in these malformed leaf blades, a few meristem tissues all can be suppressed or accelerated, and normal leaf blade morphology disappears. Pro ASL11:GUS expression further suggests that this process may be triggered by misexpression of a few transcription factors (Tfs).

A list of identified oribatid mites of the family Lohmanniidae (Acari, Oribatida) collected from Cuba, including 4 species from 4 genera, is provided. The genus Heptacarus and species Heptacarus supertrichus Piffl, 1967 are recorded in Cuba for the first time. A new species of Annectacarus is described from leaf litter in forest of Valle de Viñales National Park. Annectacarus vinalesensis sp. n. is morphologically most similar to A. krachan Mahunka, 1995, but the new species differs from the latter by the rostrum with indentation and the presence of six pairs of neotrichal notogastral setae and six pairs of setae on epimere I. An identification key to the known species of Annectacarus in the Neotropical region is given.

Papua New Guinea is one of the most valuable tropical regions but ecological research of its freshwater invertebrates has been lacking. The goal of this paper is to evaluate the species richness, diversity and structure of aquatic insect assemblages in different habitats in the Wanang River catchment in a well-preserved lowland rainforest. Assemblage structure was studied on two spatial scales - in different habitats (river, streams and stagnant pools) and in three mesohabitats in the river (slow and fast sections and submerged wood). The results show that headwater streams had the highest morphospecies diversity, while the river had the highest insect abundance. Slow and fast sections of the river differed both in terms of insect abundance and diversity. Furthermore, a number of unique wood-associated species was found on submerged wood. The most notable feature of the assemblage structure was scarcity of shredders and dominance of predators. However, predatory beetles, bugs and dragonfly larvae exhibited contrasting habitat preferences. This study shows that Papua New Guinean lowland rainforests host diverse and distinctly structured freshwater insect assemblages.

Grasshopper species belonging to subfamilies Melanoplinae, Gomphocerinae and Oedipodinae were tested for their feeding rate on three types of grass. All grasshopper species were offered Shawnee and Kanlow cultivars of switchgrass, Panicum virgatum L. and big bluestem, Andropogon gerardii Vitman. The grasshoppers, Melanoplus femurrubrum and Melanoplus differentialis were also tested for their feeding on turgid or wilted leaves of the Shawnee cultivar of switchgrass. We found that M. differentialis consumed more switchgrass compared to big bluestem while M. femurrubrum and Arphia xanthoptera consumed the most Shawnee switchgrass. The M. differentialis consumed more turgid grass compared to wilted switchgrass. The feeding performances show differences among grasshopper species even in the same subfamily and suggest that Melanoplinae grasshoppers may become destructive pests of switchgrass planted for biofuel production.

The nests of the harvest mouse (Micromys minutus) from an extensively used farmland in Western Poland were recognized as a place of occurrence of a large group of invertebrates. In total, from 42 nests 6,544 individuals of invertebrates were extracted. The number of invertebrates was positively correlated with mass of mouse nests. There were found arthropods belonging to the following groups: Arachnida: spiders, mites and pseudoscorpions; Entognatha: springtails; Insecta: Coleoptera, Diptera, Hemiptera, Hymenoptera, Lepidoptera, Psocoptera, Siphonaptera and Thysanoptera. The most numerous groups were mites (Acari, 5,800 individuals) and fleas (Siphonaptera, 415 individuals). Among mites Astigmatida, Mesostigmata and Oribatida were identified. One species, Tyrophagus curvipenis (Acari: Astigmatina), was found in Poland for the first time. The study clearly showed that nests of the harvest mouse are often inhabited by invertebrates, and looks like some of them are characteristic to this particular kind of habitat.

The aim of the study was to describe the possible anatomical variations in origin of the branches arising from the descending aorta in rabbit and hare. The study was carried out on ten adult rabbits and ten adult European hares. The study was carried out using the corrosion technique. After the euthanasia, the vascular network was perfused with saline. Spofacryl Dental © was used as a casting medium. After polymerisation of the medium, the maceration was carried out in KOH solution. We found high variability in origin of the branches arising from the descending aorta in both species. The variations in the level of the origin of the celiac artery, cranial mesenteric artery, renal arteries, ovarian arteries, testicular arteries and caudal mesenteric artery were present between both species and within the same species. In some individuals of both species, variations in the level and the arrangement of the origin of the dorsal intercostal arteries, lumbar arteries and median sacral artery were present. According to our results, it can be concluded that the anatomical arrangement of branches of descending aorta shows a higher number of variations in domesticated rabbit in comparison with the hare.

The aim of investigating protein interaction between Homo sapiens adenovirus and heat shock 70 kDa protein (Hsp70) is to study a potentially synergistic interaction that would enhance the anti-apoptotic mechanisms, hence increasing the virus replication rate and improve the killing efficiency of tumour cells in cancer therapy. Currently, the protein interaction between Hsp70 and E1A 32 kDa of human adenovirus C serotype 5 (Ad5) is still unknown. Mutant models (T11V, T12P and D364S) were built, simulated and their interactions with Ad5 were studied. The E1A 32 kDa of human Ad5 motif (PNLVP) showed the lowest binding energy and intermolecular energy values with the novel T11V mutant at -8.26 kcal/mol and -11.21 kcal/mol. The protein-ligand complex models revealed that the T11V mutant had the strongest and most stable interaction with the PNLVP motif among all the four protein-ligand complex structures. This knowledge would assist future in vivo investigations of this protein-ligand complex structure in cancer treatment research.

The pathophysiology of multiple sclerosis (MS) includes also the vascular abnormalities. Recent reports on changes in venous cerebrospinal outflow and the investigation of immunomodulatory properties of several vascular mediators on the molecular level have added new information to hypotheses on vascular pathology as determining factor in the pathophysiology of MS. We assessed changes in serum paraoxonase 1 (PON1) activities in MS patients and polymorphism of PON1 as a risk factor for MS. The main role of serum PON1 is hydrolysis of lipid peroxides and protection of lowdensity lipoprotein particles from oxidation. These events could play a role in lowering the risk of atherogenesis and vascular complications development in MS. There are controversial results about association of two main polymorphisms in paraoxonase coding region (PON1 55L/M, PON1 192Q/R) and risk of MS in different populations. Our results support studies that PON1 polymorphisms are probably not a risk factor of MS development.