Gene silencing has been recognized as an important factor, which influence the expression of heterologous protein delivered via plant-viral based system. The application of suppressor genes that act against gene silencing mechanisms demonstrated improvement in the expression level of heterologous protein in plants. The aim of the present study was to evaluate the ability of a silencing suppressor gene to enhance the expression of VP2 protein of highly virulent infectious bursal disease virus in Nicotiana benthamiana plants using a plant viral-based expression system derived from potato virus X. In the present study, a silencing suppressor gene of P19 encoded by tomato bushy stunt virus was employed as a suppressor for gene silencing mechanism by co-infecting N. benthamiana plants with the P19 gene via agroinfiltration strategy followed by mechanical inoculation of potato virus X viral transcripts containing the cloned VP2 gene. Western blot analyses revealed that the expressed VP2 protein can be detected by 7 days post-infection in the presence of P19 gene as compared with plants without P19 gene that can only be detected at 10 days post-infection. However, there was no significant enhancement in the yield of expressed VP2 protein in the presence of P19 gene. The present findings demonstrated the employment of P19 gene and improved the expression of hvIBDV VP2 protein in N. benthamiana plants.

Vegetation influences the stability of slope by both hydrological events and mechanical reinforcement of the soil. The magnitude of such effects is subject to the root system growth, which in turn is a purpose of the genetic properties of the species and site appearances. In this study, we investigated the root distribution of Macaranga tanarius (L.) Mull. Arg. as a native species in Malaysia, with an aim to rise our knowledge on root area ratio distribution inside the soil. Concerning the estimation of root distribution within the soil, we assessed the root area ratio (the proportion between the area occupied by the roots in a slice area of soil) according to its depth for four samples in the rainforest. Results show that the root area ratio (RAR) declined with depth, and the higher RAR values were calculated in the higher layers. RAR values were between 0.950% and 18.477%. There is a plentiful diversity of root density in depth classes. Also, the result showed that about 50% of the roots are located in the first 10 cm layer, and about 87% of roots were in the fine root diameter (> 2 mm). There is not a significant difference between RAR and soil depth. Spearman correlation showed no significant and negative correlation between RAR and depth. The maximum RAR percentage was in the first layer 0-10 cm (44.59%). Also, the results showed that the amount of RAR decreased with depth in fine roots (d < 2 mm), but RAR in thin roots (2 < d < 10 mm) has an unexpected change. Also, the fine root number was much higher than the number of thin roots, but the RAR value in thin roots was much higher than of the fine roots.

Antimicrobial peptides are the upcoming therapeutic molecules as alternative drugs to the existing antibiotics owing to their potent action against pathogenic microorganisms. In this study, to obtain an antimicrobial peptide with a broad range of activity, the synthetic cationic antimicrobial peptide was designed by using in silico tools viz., antimicrobial peptide database, protparam, hierarchical neural network. Later, the peptide was translated back into a core nucleotide sequence and the gene for the peptide was constructed by overlapping PCR. The amplified gene was cloned into pRSET–A vector and transformed into salt inducible expression host E. coli GJ1158. The expression results show high yields of soluble recombinant fusion peptide (0.52 g/L) from salt-inducible E. coli. The recombinant peptide was purified by the IMAC purification system and cleaved by enterokinase. The digested product was further purified and 0.12 g/L of biologically active recombinant cationic antimicrobial peptide was obtained. In vitro analysis of the purified peptide demonstrated high antimicrobial activity against both Gram positive and Gram negative bacteria devoid of hemolytic activity. Therefore, this synthetic cationic antimicrobial peptide could serves as an promising agent over chemical antibiotics. In this study, a synthetic cationic antimicrobial peptide was designed, cloned and expressed from salt-inducible E. coli GJ1158 using cost effective media in the large scale production of antimicrobial peptide and its biological activity was analysed against different Gram positive and negative organisms.

This study describes an adaptation of the Comet assay under neutral conditions for mono- and dicotyledonous plants pea (Pisum sativum L.) and wheat (Triticum aestivum L.). Modifications concern lysis and electrophoresis steps, respectively. Electrophoresis was carried out varying the intensity of the electric field. A linear relationship between the percentages of DNA in the tail from control background with alteration of intensity was found. Trypan blue dye exclusion test was used in order to determine the intactness of nuclear membrane of the isolated nuclei from both plant model systems. Assessment was conducted on non-irradiated and irradiated nuclei on a monolayer with three doses of UVC. It was found that the share of intact nuclei (trypan blue negative ones) is about 95% in controls. Gradual dose-related increase of damaged nuclei was observed in both species, reaching statistical significance only at the higher dose applied.

YKL-40 is a member of mammalian chitanase (CHI3L1), expressed and secreted by several types of solid tumor cells, inflammatory cells and stem cells. The precise physiological role of YKL-40 in cancer is still not clear and it is suggested that it play a role in cancer cell proliferation, differentiation, metastatic potential, cell attachment and migration, reorganization and tissue remodeling. The aim of the study was to check the appearance of YKL-40 in leukemic cells and over-expression of YKL-40 in the plasma of leukemia patients in comparison to healthy controls, and find whether YKL-40 could serve as a peripheral biomarker for leukemia. The study was conducted between July 2012 and March 2013 and included 67 volunteers, 55 having leukemia at the stage of diagnosis of the disease and 12 normal healthy volunteers. YKL-40 levels were determined in all plasma samples using the YKL-40 enzyme-linked immunosorbent assay (ELISA) kit and expression of YKL-40 was observed by using immunocytochemical (ICC) analysis. YKL-40 plasma levels differed significantly between patients with leukemia and the normal healthy volunteers (P=<0.001) and YKL-40 was positively expressed in all four types of leukemia (AML, ALL, CLL and CML) specimens.

Hyaluronic acid (HA) is a hydrated gel and comprises repeating units of glucuronic acid and N-acetylglucosamine. Production and recovery of HA has gained great importance due to its vast clinical applications. In pursuit of obtaining highly pure HA, we have developed a fed-batch fermentation process using Streptococcus zooepidemicus in a 25 L bioreactor that resulted in a maximum yield of 2.3 g/L HA. In addition, we have devised an efficient method for separation and recovery of hyaluronic acid from a highly viscous broth by treating with trichloroacetic acid (0.1%) and charcoal (1-2%), passing through filtration (0.45 μm) and ultrafiltration that resulted in recovery of 72.2% of clinical grade HA with molecular weight of 2.5x106 Da. We have also characterized our purified HA using FTIR and NMR spectroscopy. These studies revealed the similarity in both the FTIR spectrum as well as NMR spectrum of both reference standard and purified HA from S. zooepidemicus indicating that the reported process is more efficient in terms of better yield and high quality (99.2%).

Conformational changes play an important role in the function of proteins. Glutamine synthetase, an important enzyme of nitrogen metabolism, was purified under sporulating (GSala) and non-sporulating (GSpyr) conditions and the effect of Mg2+ on these multiple forms was studied by fluorescence spectroscopy to detect possible conformational changes that occur in the presenceof Mg2+. The substantial changes in the fluorescence emission maximum, fluorescence intensity and lifetime that occur in the presence of different concentrations of Mg2+, indicated major changes in molecular conformations in both forms of this enzyme. The fluorescent changes produced by the effect of Mg2+ in GSala was much more prominent than in GSpyr. These observations strongly support the possibility that GSala and GSpyr undergoes a conformational change on binding with Mg2+.

Genetic diversity and relationships among six rice genotypes were investigated using five random amplified polymorphic DNA (RAPD) markers. A total of 69 alleles were amplified, of which 66 were polymorphic. The size of the amplified alleles was between 0.25 and 2.35 kbp. The number of polymorphic alleles detected with each primer ranged from 7 to 24 with an average of 13.2 per primer and the polymorphism information content (PIC) values varied from 0.8672 to 0.9471. Pair-wise similarity estimated the range of 0.308 to 0.718 among all the genotypes and the highest genetic similarity was found between Maloti and BRRI dhan53. Cluster analysis using UPGMA (unweighted pair group method with arithmetic averages) revealed three clusters at genetic similarity of 46%. A moderately drought tolerant landrace, Boalia, formed a single cluster and the remaining genotypes grouped into distinct clusters based on their relatedness. The results showed a high level of genetic diversity among studied genotypes and this information will assist in conservation as well as selection of parents during breeding programs for the development of drought tolerant rice varieties in near future.

The essential oils obtained by hydrodistillation of the flowers of Santolina africana Jord. et Four. were analyzed by GC/MS. The main constituents of the essential oil were acenaphtane (25.23 %), calarene (21.54 %), ocimene (17.44 %) and some other compounds that were presented only in minor amounts. In total, essential oils of Santolina africana Jord. et Four. were considered as a rich source of hydrocarbon mono and sesquiterpenes. Moreover, the antimicrobial activity of the essential oil against eight bacteria strains and three fungi was studied. It was found that the most powerful effect was against Bacillus subtilis, Staphylococcus aureus and Aspergillus flavus. The essential oils were screened for their possible in vitro antioxidant activity by DPPH free radical-scavenging test. The findings showed that reduction percentage is 13.80 at 1M.

The aim of the study was to assess the effects of dietary supplementation with two potential probiotics, Bacillus subtilis and Bacilis cereus toyo, on selected biomarkers, not directly related to nutrition, in rainbow trout Oncorhynchus mykiss (Walbaum, 1792). A nine week-long diet with the probiotics was found to affect hepatic EROD and GPx; muscular AChE, BChE, POx and LP; and branchial Na+/K+-ATPase, as compared to an equal diet without probiotics supplementation. No effects on hepatic GST, SOD, CAT, POx, LP, LPO, PY; muscular GPx, LPO; brain AChE, BChE; and gross morphometric indices were found.