Baqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11864220170601Novel PCR-ELISA Technique as a Good Substitute in Molecular Assay56767268984ENFatemehTayebehDepartment of Biology, Damghan Azad University, Damghan, IranShahramNazarianDepartment of Biology, Faculty of Science, Imam Hossein University, Tehran, IranSeyed AliMirhosseiniApplied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, IranJafarAmaniApplied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran0000-0002-5155-4738Journal Article20170611Due to the spread of infectious diseases, the existence of a rapid and sensitive detection method is necessary today. Polymerase chain reaction-enzyme linked immunosorbent assay (PCR-ELISA) is a simple manner for detection of microorganism. For example, bacteria, viruses, fungi and others based on nucleic acid sequence. A large number of samples can be screened by this technique simultaneously, so it is not time consuming and is a quick manner. The high sensitivity and specificity of PCR-ELISA make it a powerful technique by simple laboratory facilities. As a result it can be an excellent substituted manner for analysis and detectionin different various fields.https://www.biotechrep.ir/article_68984_72ef19939d164082d123f0dea7638bae.pdfBaqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11864220170601Leaching Characteristics of Phenanthrene and Pyrene in Biochar-amended Contaminated Soil57358168986ENAliDaryabeigi ZandGraduate Faculty of Environment, University of Tehran, Tehran, Iran0000-0002-2490-0893Journal Article20170411Contamination of soil with persistent organic contaminants has been of great concern due to their long-term effects in the environment. Polycyclic aromatic hydrocarbons (PAHs) are a well-known group of organic contaminants characterized by their resistance to biodegradation and high hydrophobicity. Mobilization and migration of PAH compounds in hydrocarbon-contaminated site may endanger groundwater resources. The main objective of this study was to investigate the influence of pulverized biochar on immobilization and release characteristics of model PAHs. Column leaching test was used to simulate leaching of PAH compounds from contaminated solid phase towards aqueous phase. Results showed strong sorption of both studied PAHs i.e. phenanthrene and pyrene to soil particles, however, at the end of the experiment 5.32% and 0.99% of the initial solid phase content of, respectively, phenanthrene and pyrene released into water in unamended soil. Application of pulverized biochar could finally reduce mobilization and cumulative release of the above-mentioned PAH compounds significantly. Similar trend was also obtained for sum 16 US EPA PAHs. Variation of pH during the leaching process and its contribution to mobilization and release of selected PAH compounds, which has scarcely studied previously, were also addressed. Results indicated that pulverized biochar as a cost-effective alternative to other carbon-rich amendments e.g. activated carbon can be effectively employed for site remediation purposes to reduce mobilization of PAHs in soil.https://www.biotechrep.ir/article_68986_fa4e665aefc7d576b6b85358fb35b60d.pdfBaqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11864220170601The Effect of Ramadan Fasting on Biochemistry of Saliva58358668987ENNarjesKhaleghifarDepartment of Biology, University of Guilan, Rasht, IranReyhanehSaririDepartment of Biology, University of Guilan, Rasht, IranMahmoodrezaAghamaaliDepartment of Biology, University of Guilan, Rasht, IranHoseinGhafooriDepartment of Biology, University of Guilan, Rasht, IranJournal Article20170411Muslims fast every day from down to sunset during the holy month of Ramadan. The possible side effect of Ramadan fasting on general health has been widely studied, but oral health may also alter due to one month fast affecting some of the salivary secondary metabolites. The aim of present research was identifying the influence of fasting on saliva of healthy individuals. The subjects were selected from non-smoker male employees of one factory in Rasht. 35 healthy male (aged 30-50 years), who fasted the whole Ramadan entered. Their unstimulated saliva samples were collected before, during and at the end of Ramadan. Concentration of salivary uric acid, the activity of alkaline phosphatase (ALP) and aspartate aminotransferase (AST) were then measured using spectrophotometric methods according to procedure given for serum. The results showed a significant reduction in the concentration of salivary uric acid and AST, while the activity of ALP was significantly increased. It was concluded that some of salivary biochemical markers undergo various fluctuations in response to fasting. However, more studies with a larger population and various biochemicals are required to certainly comment on the matter.https://www.biotechrep.ir/article_68987_d4fa68c995981c4ffa9aff9a4b7a1287.pdfBaqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11864220170601Application of Microbial Biotechnology in Conservation and Restoration of Stone Monument58759268992ENNargesZamaniRestoration of historical and Cultural objects, Isfahan Art University, Isfahan, IranMohsenGhezelsoflaIranian Blood Transfusion Organization, Shahr-e-kord, IranAli MohammadAhadiDepartment of Biology, University of Shahrekord, Shahrekord, IranMarziyeZamaniIranian Blood Transfusion Organization, Shahr-e-kord, IranJournal Article20170411Treatments employed for the consolidation of monumental stones made of limestone due to incompatibility from the substrate and cement used for consolidation, plugging of pores induced by the new cement, leading to the acceleration of stone alteration. Microbial precipitation with a layer of calcium carbonate generated by bacteria might offer a solution to this dilemma because the layer would not block the natural pore structure, thus permitting free passage of soluble salts through the stone. In this study, an attempt has been made to provide an overview of the microbial induced carbonate precipitation as promising technology for bioremediation of such structures. At the first, the active microorganisms in the conservation of stone monuments transferred to the laboratory using the swap dipped in nutrient broth at a historic cemetery. After incubation and growth of colonies, Gram-positive bacilli were detected. Then pure single colonies were transferred to blood agar medium and incubated at 37°C. The single colonies were transferred to the surface of sterilize limestone pieces and incubated but no result was obtained. Therefore, in the next phase bacilli bacteria-rich broth media was used. The control experiments were conducted in accordance with the conditions mentioned without bacterial inoculation. The calcification process caused by the inoculated bacteria on the historical stone samples was demonstrated using the scanning electron photomicrographs. Microbially induced calcium carbonate precipitation (MICP) technology to eco–friendly, self-healing and highly durable nature of these bio-binders, for conservation purposes has been found suitable. But still there has been much to explore in order to bring this environmentally safe, cost effective and convenient technology from lab to field scales.https://www.biotechrep.ir/article_68992_0fb813a400dd07c7693c534929c1a4c2.pdfBaqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11864220170601In Silico Analysis of a Multi-subunit Immunogen, Targeting Virulence Factors of Enterohemorrhagic Escherichia coli59360269002ENEmadKordbachehDepartment of Biological Sciences,, Faculty of Sciences, Imam Hossein University, Tehran, IranShahramNazarianDepartment of Biological Sciences,, Faculty of Sciences, Imam Hossein University, Tehran, IranMiladAmerianDepartment of Biological Sciences,, Faculty of Sciences, Imam Hossein University, Tehran, IranJournal Article20170511Enterohemorrhagic <em>Escherichia coli</em> (EHEC) strains are foodborne pathogens with importance in public health. The lack of effective clinical treatment, sequelae after infection and mortality rate in humans confirms the essential need for prophylactic and vaccines approaches. EspA, Tir, HcpA and Stx2 are major virulence factors for adherence and toxicity of EHEC, so an appropriate tetravalent immunogen consist of toxin subunit and crucial colonization factors was selected and constructed. Bioinfo[1]rmatic analyses of recombinant construction such as sequences choosing and optimizing, mRNA folding, physicochemical property in 2D and 3D structures, besides other immunoinformatics data like B-cell and T-cell epitopes and allergenicity of chimera were some reported according to the reliable servers. <em>In silico</em> assessment of the chimeric proteins demonstrated the desired model has a proper mRNA features, besides acceptable stability and solubility. This model is close to native proteins topologically, and all domains were found to have a high antigenic competency and surface accessibility. These results can be beneficial for the development of a chimeric immunogen against adherence and toxicity of EHEC in an animal model application.https://www.biotechrep.ir/article_69002_2aee7f47c0321317cd6ebb336d467080.pdfBaqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11864220170601Fabrication of an Amperometric Glucose Biosensor Based on a Prussian Blue/Carbon Nanotube/Ionic Liquid Modified Glassy Carbon Electrode60360869004ENShararehSajjadiDepartment of Biology, Roudehen Branch, Islamic Azad University, Roudehen, IranAmir HomayounKeihanMolecular Biology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, IranParvizNorouziCenter of Excellence in Electrochemistry, Faculty of Chemistry, University of Tehran, Tehran, IranMohammad MahdiHabibiCenter of Excellence in Electrochemistry, Faculty of Chemistry, University of Tehran, Tehran, IranKhadijehEskandariNanobiotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IranNajmehHadizadeh ShiraziDepartment of Biology, Roudehen Branch, Islamic Azad University, Roudehen, IranJournal Article20170411An amperometric glucose biosensor was developed based on synergistic contributions of prussian blue (PB) and a bucky gel (BG) consisting of carbon nanotubes (CNTs) and ionic liquid (IL). The PB nanoparticles were first deposited onto the surface of a BG modified glassy carbon (GC) electrode (BG/GC). Then, the Ni<sup>2+</sup> ions were electrochemically inserted into the PB lattice to improve its stability at physiological pH. Afterwards, Glucose oxidase (GOx) was immobilized on the BG/GC electrode using a cross-linking method. Amperometric measurements of glucose were performed at −0.05 V vs. Ag/AgCl in 0.05 M phosphate buffer solution at pH 7.4. The glucose biosensor exhibited a sensitivity of 45.03 µA/(mM.cm<sup>2</sup>) with a detection limit of 5×10<sup>-7</sup> M. The amperometric response was linear in the range of 5×10<sup>-7</sup> to 8.3×10<sup>−4</sup> M.https://www.biotechrep.ir/article_69004_cf2d8d4cb40c63ccfd1336f48ba0408d.pdfBaqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11864220170601A New and Efficient Method for DNA Extraction from Human Skeletal Remains Usable in DNA Typing60961469005ENAliMohammadiHuman Genetics Research Center, Baqiyatallah University of Medical Sciences Tehran, IranAkbarGhorbani AlvaneghHuman Genetics Research Center, Baqiyatallah University of Medical Sciences Tehran, IranYoung Researchers and Elite Club, Islamic Azad University, Tehran, IranMostafaKhafaeiHuman Genetics Research Center, Baqiyatallah University of Medical Sciences Tehran, IranSajjadHabibi AzarianHuman Genetics Research Center, Baqiyatallah University of Medical Sciences Tehran, IranMahdiNaderiHuman Genetics Research Center, Baqiyatallah University of Medical Sciences Tehran, IranEbrahimKiyaniHuman Genetics Research Center, Baqiyatallah University of Medical Sciences Tehran, IranAliMiriHuman Genetics Research Center, Baqiyatallah University of Medical Sciences Tehran, IranHasanBahmaniHuman Genetics Research Center, Baqiyatallah University of Medical Sciences Tehran, IranMaryamRamezaniHuman Genetics Research Center, Baqiyatallah University of Medical Sciences Tehran, IranMahmoodTavallaeiHuman Genetics Research Center, Baqiyatallah University of Medical Sciences Tehran, IranJournal Article20170611In historical cases, mass disasters, missing person’s identification and ancient DNA investigations, bone and teeth samples are often the best and the only biological material available for DNA typing. This is because of the physical and chemical nature of the protein-mineral matrix of bone which is relatively resistant to the adverse environmental effects and biological attacks. Most bone extraction protocols used in the forensic laboratories involve an incubation period of bone powder in extraction buffer for proteinase digestion, followed by the collection of the supernatant, and the elimination of large quantities of undigested bone powder. Here we demonstrate an extremely effective protocol for recovery of DNA. This is performed in a method that retains and concentrates all the reagent volume for complete DNA recovery. For our study, we selected challenging bone samples of skeleton remains of the martyred individuals in Iraq’s imposed war on I.R. Iran (1980-1988). The bones that were extracted with our new protocol showed that this new protocol significantly enhances the quantity of DNA that can be used for amplification from degraded skeletal remains. At the same time we tested in parallel the samples by using of QIAamp DNA Investigator Kit and attained the best results by using new protocol. In fact, our new DNA extraction method is based on previous standard methods such as Chelex and salting out. We have used this technique to successfully recover authentic DNA Typing from extremely challenging samples that failed repeatedly using the standard protocols. However, the amount of recovered DNA was very small but it was possible to extract genomic DNA from these challenging bone samples. The results indicated that our procedure for DNA extraction although yielded little amount of genomic DNA; however, it was pure DNA that can be used for further analysis such as PCR amplification and DNA profiling. Since the new procedure is fast and needed less time than previously standard procedures, we have named it FDEB (Fast DNA Extraction of Bone).https://www.biotechrep.ir/article_69005_a5db12b8170a5c8a02c70d183784aaa4.pdf