Journal of Applied Biotechnology Reports
Volume:2 Issue: 2, Spring 2015

Daily, organophosphorus compounds (OPs) in human life, has found wide applications. Although OPs have biodegradability potential, they induce clinical problems in humans and other organism. Different methods are used to detoxify these compounds. In the meantime, biodegradation is preferred as a compatible way to the environment since it produces less toxic compounds. Enzymes capable to degrade the OPs are of the most important items in the biodegradation. Genetic manipulation involved in the production of these enzymes has been employed in bacteria, and finally, is used for the mass production of recombinant microorganisms. In this paper, the role of organophosphates on human life and the ways to destroy toxic organophosphates are studied.

Millions of diarrheal disease is made by Enterotoxigenic E. coli (ETEC) each year, specifically in developing countries. In the pathogenesis of ETEC infections, the first phase is sticking of the bacterium to the minute intestinal epithelium, as a result of colonization factors (CFs) mediation and subsequently generate enterotoxins. These CFs in accordance with their structure are diverged into discrete groups. CFA/I and CS6 are two of the most typical CFs. CFA/I is a fimbriae consists of a superior subunit, CfaB and inferior subunit, CfaE. CS6 is non-fimbrial which includes two main subunits, CssA and CssB. The enterotoxins caused by ETEC are related to two eminent classes of heat-labile toxins (LT) and heat stable toxins (ST). LT is formed of five B subunits and a single enzymatically active A. Its B subunits tied up to the enteral GM1 ganglioside receptors in the intestinal epithelium and A subunit whose ADP-ribosylating activity culminates in cellular adenylcyclase activation and an increase in cAMP, efflux of chloride ions and water and succeeding watery diarrhea. Guanylatecyclase (GC) is receptor for the ST toxin. Intracellular levels of cyclic guanosine monophosphate (cGMP) increase when ST binds to GC. Such increase in cGMP permits activation of cystic fibrosis transmembrane conductance regulator (CFTR) by phosphorylation-dependent cGMP protein kinase II producing an escalation in salt and secretion of water and prevention of sodium absorption through h the apical Na/H channel.More information about the CFs and enterotoxins of pathogen leads to more founding of ETEC virulence, and the founding is important to designing an appropriate vaccine.

Influenza A virus is now considered to be widespread in poultry and has demonstrated the ability to infect humans in Iran. For laboratory diagnosis of these respiratory viruses, it is essential to have rapid methods, able to detect viruses in early stages of the infection in clinical specimens. The real-time reverse-transcription polymerase chain reaction n (rRT-PCR) assay has been established as a standard method for Influenza virus type A diagnosis inpatients. In this study, we evaluated a single-tube rRT-PCR assay, targeting to the highly conserved region of matrix (M) gene for detection of the virus. In this experimental study, after preparation of 100throat mucus samples, respective RNA was extracted from the virus by using viral RNA extraction kit. Two specific primers were synthesized, based on the conserved region of Influenza type AM-gene and a home-brewed one-step SYBR Green based rRT-PCR was developed and evaluated for detection of Influenza type A infection in the viral samples, on the basis of melting curve analysis. The presence of M-gene in RNAs, extracted from 53viralsamples, was confirmed by this single-tube rRT-PCR assay, and after 45 amplification cycles, the melting curve analysis revealed the melting temperature (Tm) of 83.2 ± 0.5°C for various viral samples, quite different from that of primer-dimers and the positive samples showed only a small variation in parameters. This study showed that the developed one-step rRT-PCR assay is the proper molecular method for rapid and accurate diagnosis of Influenza A by detection of M-protein encoding gene.

The aggregation of protein is the most prevalent and the most disturbing kind of instability and this challenge exists in almost every stage of the development of protein drug. The presence of insoluble aggregations in protein drugs will make the supply of the product a tough job. This study identifies the inhibition of the folded Interferon beta 1-b’s aggregation with the assistance of some excipients. It uses some thermal stress and mechanical methods to accelerate the aggregation, and also the spectroscopic method to identify the protein aggregation and its growth. Experimental data of the tests show compliance with the autocatalytic model. This model has been used to obtain the Kinetic constants of aggregation in different states and to make comparison with one another in the presence of some excipients. The kinetic constants were obtained by fitting the Autocatalytic model on data. Among these excipients, Polysorbate 20 of 0.01% (w/v) showed the best result in decreasing the aggregation. Using this excipient of 0.01% (w/v) in thermal stress causes dramatic reduction of nucleation constant from 8.3× 10-3 (min-1) to 4.14× 10-6 (min-1), which indicates the reduction of protein aggregation in the solution.

Chickpea (Cicer arietinum L.) as a member of the Legumine family is one of the most important plant protein resources. One of the methods for increasing of variation is somaclonal variation. The most common factors affecting somaclonal variation are explant and growth regulators in which the culture is established. The current research was setup as a callus induction experiments for comparison of different growth regulators and explants of chickpea (Bivanij cultivar). The callus induction experiment was conducted in MS medium supplemented by different concentrations of 2, 4-D (0, 1, 2 and 4 mg/l) plus 0.1 m/l BAP in five explants (embryo, seed, root, hypocotyl and cotyledon). The evaluated traits include days to callus induction, callus induction percentage and callus growth rate. Results showed that mostly medium induced the callus formation and the best response was in medium supplemented by 2 mg/l 2, 4-D. The lowest days to callus induction was belong to embryo explant in medium supplemented by 0.1mg/l BAP+1mg/l 2,4-D (4.25 days) and highest days to callus induction was belong to cotyledon explant in medium supplemented by 0.1mg/l BAP+2 mg/l 2,4-D (14.25 days). The highest callus growth rate was belong to embryo in medium supplemented by 1 mg/l 2, 4-D (0.1775 mm diameter/d) and the lowest was belong to cotyledon (0.02 mm diameter/d) in 4 mg/l 2, 4-D and seed (0.01mm diameter/d) in 1 mg/l 2, 4-D.

In this study, the substrate diffusion in an immobilized spherical cell-support aggregate is studied and effects of various parameters are investigated on substrates profile. Analyses are performed by using of an analytical solution called the Least Square Method (LSM) and results are compared with numerical solution. The effects of effective diffusion coefficient (De), maximum specific growth rate (µm) and type of limiting substrate are studied on substrate concentration profile in immobilized Nitrosomonas europaea and Nitrobacter agilis microorganisms. Outcomes reveal that LSM is an appropriate method for analyze of biological non-linear equations. In the center of the spherical microbial support, the substrates concentration is minimums and with reducing µm or increasing De, substrate concentration profile gradient reduces.

The alginate was extracted with six different methods from Iran south seacoast algae, Sargassum sp, and the percentage yield of alginate was determined. We divided our methods to two groups including acidic extraction and non-acidic. In acidic methods, HCL and H2SO4 were used as a detergent in extraction process and CaCl2 was exerted in non-acidic treatments. All treatments compared with each other and indicated an increasing in alginate yield when different methods used EDTA in extraction process. Finally, the main characteristics of sodium alginate were realized with FT-IR and H-NMR.