Ghosh, Prithwi; Roy, Amit; Chakraborty, Joydeep; Das, Sampa
Genetic engineering has established itself to be an important tool for crop improvement. Despite the success, there is always a risk of food allergy induced by alien gene products. The present study assessed the biosafety of mutant Allium sativum leaf agglutinin (mASAL), a potent antifungal protein generated by site directed mutagenesis of Allium sativum leaf agglutinin (ASAL). mASAL was cloned in pET28a+ and expressed in E. coli, and the safety assessment was carried out according to the FAO/WHO guideline (2001). Bioinformatics analysis, pepsin digestion, and thermal stability assay showed the protein to be nonallergenic. Targeted sera screening revealed no significant IgE affinity of mASAL. Furthermore, mASAL sensitized Balb/c mice showed normal histopathology of lung and gut tissue. All results indicated the least possibility of mASAL being an allergen. Thus, mASAL appears to be a promising antifungal candidate protein suitable for agronomical biotechnology.
Burger-Helmchen, Thierry, Ed.
Chapters in this book include: (1) The Psychology of Entrepreneurship (Melek Kalkan and Canani Kaygusuz); (2) Entrepreneurial Intentions: The Role of the Cognitive Variables (Jose C. Sanchez); (3) Do External Factors Influence Students' Entrepreneurial Inclination? An Evidence Based Approach (Ishfaq Ahmed, Muhammad Musarrat Nawaz and Muhammad…
Banerjee, Nilanjana; Ghosh, Prithwi; Das, Kalipada; Das, Sampa
Background Allium sativum leaf agglutinin (ASAL) is a 25-kDa homodimeric, insecticidal, mannose binding lectin whose subunits are assembled by the C-terminal exchange process. An attempt was made to convert dimeric ASAL into a monomeric form to correlate the relevance of quaternary association of subunits and their functional specificity. Using SWISS-MODEL program a stable monomer was designed by altering five amino acid residues near the C-terminus of ASAL. Methodology/Principal Findings By introduction of 5 site-specific mutations (-DNSNN-), a β turn was incorporated between the 11th and 12th β strands of subunits of ASAL, resulting in a stable monomeric mutant ASAL (mASAL). mASAL was cloned and subsequently purified from a pMAL-c2X system. CD spectroscopic analysis confirmed the conservation of secondary structure in mASAL. Mannose binding assay confirmed that molecular mannose binds efficiently to both mASAL and ASAL. In contrast to ASAL, the hemagglutination activity of purified mASAL against rabbit erythrocytes was lost. An artificial diet bioassay of Lipaphis erysimi with mASAL displayed an insignificant level of insecticidal activity compared to ASAL. Fascinatingly, mASAL exhibited strong antifungal activity against the pathogenic fungi Fusarium oxysporum, Rhizoctonia solani and Alternaria brassicicola in a disc diffusion assay. A propidium iodide uptake assay suggested that the inhibitory activity of mASAL might be associated with the alteration of the membrane permeability of the fungus. Furthermore, a ligand blot assay of the membrane subproteome of R. solani with mASAL detected a glycoprotein receptor having interaction with mASAL. Conclusions/Significance Conversion of ASAL into a stable monomer resulted in antifungal activity. From an evolutionary aspect, these data implied that variable quaternary organization of lectins might be the outcome of defense-related adaptations to diverse situations in plants. Incorporation of mASAL into agronomically
Bashapoor, Sajjad; Hosseini-Kiasari, Seyyedeh Tayebeh; Daneshvar, Somayeh; Kazemi-Taskooh, Zeinab
Background Sensory information processing and alexithymia are two important factors in determining behavioral reactions. Some studies explain the effect of the sensitivity of sensory processing and alexithymia in the tendency to substance abuse. Giving that, the aim of the current study was to compare the styles of sensory information processing and alexithymia between substance-dependent people and normal ones. Methods The research method was cross-sectional and the statistical population of the current study comprised of all substance-dependent men who are present in substance quitting camps of Masal, Iran, in October 2013 (n = 78). 36 persons were selected randomly by simple randomly sampling method from this population as the study group, and 36 persons were also selected among the normal population in the same way as the comparison group. Both groups was evaluated by using Toronto alexithymia scale (TAS) and adult sensory profile, and the multivariate analysis of variance (MANOVA) test was applied to analyze data. Findings The results showed that there are significance differences between two groups in low registration (P < 0.020, F = 5.66), sensation seeking (P < 0.050, F = 1.92), and sensory avoidance (P < 0.008, F = 7.52) as a components of sensory processing and difficulty in describing emotions (P < 0.001, F = 15.01) and difficulty in identifying emotions (P < 0.002, F = 10.54) as a components of alexithymia. However, no significant difference were found between two groups in components of sensory sensitivity (P < 0.170, F = 1.92) and external oriented thinking style (P < 0.060, F = 3.60). Conclusion These results showed that substance-dependent people process sensory information in a different way than normal people and show more alexithymia features than them. PMID:26885354
Korost, Dmitry; Gerke, Kirill; Akhmanov, Grigory; Vasilyev, Roman; Čapek, Pavel; Karsanina, Marina; Nadezhkin, Dmitry
Hydrocarbons (HC) are generated from solid organic matter (kerogen) due to thermocatalytic reactions. The rate of such reactions shows direct correlation with temperature and depends on the depth of source rock burial. Burial of sedimentary rock is also inevitably accompanied by its structural alteration owing to compaction, dehydration and re-crystallization. Processes of HC generation, primary migration and structural changes are inaccessible for direct observation in nature, but they can be studied in laboratory experiments. Modern technical facilities of laboratories make it possible to carry out experiments on HC generation from the organic-rich rocks at a completely new level (Kobchenko et al., 2011). Some new technologies, including X-ray microtomography and pore-scale modeling, allow us to carry out a step-by-step description of such processes and their development, and to study their reflection in alterations of rock structure. Experiments were carried out with a clayey-carbonate rock sample of the Domanic Formaition taken at a depth of 1939 m from borehole drilled in the central part of the Melekes depression (West Tatar arch, Russia). The rock chosen fits the very essential requirements for studying HC generation under laboratory conditions - high organic matter content and its low metamorphic grade. Our work aimed such a study in an undisturbed rock sample by heating it in nitrogen atmosphere based on a specified temperature regime in a RockEval6 analyzer and monitoring alterations in the pore space structure. Observations were carried out with a SkyScan-1172 X-ray microtomography scanner (resulting scan resolution of 1 µm). A cylinder, 4 mm in diameter, was prepared from the rock sample for the pyrolitic and microtomographic analyses. Scanning procedures were carried out in 5 runs. Temperature interval for each run had to match the most important stage of HC generation in the source rock, namely: (1) original structure; (2) 100-300˚? - discharge of