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Journal of Applied Biotechnology Reports

Journal Papers (38) Details Call for Paper Manuscript submission Publication Ethics Contact Authors' Guide Line
1 A Review on Engineering of Organophosphorus Hydrolase (OPH) Enzyme, Gholamreza Farnoosh, Ali Mohammad Latifi
Organophosphorus chemicals are compounds which have been used as pesticides and insecticides in agriculture. Theyre also used as nervous agents and have raised many problems for human and environment. Among the most important methods of decontamination from these compounds are biodegradation methods. Using OPH enzyme in degradation the mentioned compounds is seen as one of the desirable ways, but low activity and specification and low thermostability are among factors significantly decreasing the optimal application of this enzyme. Using methods of protein engineering based on the alteration of specific protein positions in order to improve the activity, specification and thermostability are some common ways used currently. Numerous studies have been done to increase activity and thermostability of OPH enzyme with alteration of some special amino acids the result of which was an increase against different substrates. OPH enzyme active site connected to substrates that consisted of three large, small and releasing packets were one of the goal areas of changing amino acids used by researchers to improve engineered activities. Among other ways of making enzymes more rigid and stable were bending loops by replacing Proline, creating disulfide bonds, ionic bonds by replacing charged amino acids.
2 Effects of missense R84Q mutation on human Pyrroline-5-carboxylate synthase enzyme properties, an in-silico analysis, Maryam Zare, Faranak Hadi, Zarrin Minuchehr, Jafar Amani, Ali Hatef Salmanian
Mammalian ?-(1)-Pyrroline-5-carboxylate synthase (P5CS) enzyme catalyzes the coupled phosphorylation and reduction-conversion of glutamate to ?-(1)-pyrroline-5-carboxylate (P5C), a critical step in the proline, ornithine, citrulline and arginine biosynthesis. In plants and mammals, P5CS consists of two separate enzymatic domains: N-terminal ?-glutamyl kinase (?-GK) and C-terminal ?-glutamyl phosphate reductase (?GPR). Hyperammonemia has been reported as a new inborn disorder, with a range of clinical symptoms which is associated with a reduced synthesis of proline, ornithine, citruline and arginine. A missense mutation, R84Q, which alters the conserved residue in ?-GK domain, is responsible for this disorder. In this study using in-silico approaches as a new bioinformatics method, sequence analysis was performed and the tertiary structure of ?-GK domain of human P5CS, which includes the R84Q missense mutation, was predicted and the mutation effects on structural and functional features of P5CS enzyme were analyzed. Our analysis showed that this substitution has an affect on the molecular surface accessibility and total energy of the modeled structure. We conclude that this mutation results in a reduced activity of P5CS enzyme and an impaired synthesis of these amino acids.
3 Improvement of maize (Zea mays L.) anther culture embryogenesis and direct regeneration by different plant growth regulators and gelling agent, Seyedeh Zahra Hosseini, Ahmad Ismaili, Payam Pour Mohammadi
Androgenesis via anther culture or microspore culture is one of the current methods for producing haploid and double haploid plants in maize. To use of this method in maize breeding program it should be able to regenerate enough plantlets. Anthers culture usually is carried out indirectly via callus induction and regeneration on at least two different media. In this study a responsive genotype, ETH-M82, using a new single culture medium was used for embryogenesis and regeneration. We tested different growth regulators (2,4-D; kinetin; NAA & IAA) in modified YP medium. After6-week, direct regeneration on some treatments was observed. The highest frequency of direct formation of plantlets (in 100 anthers) occurred on medium supplemented with 2mgl-1 IAA and 2mgl-1 kinetin (4%).Best results with an average of 3.1plantlets in replication were obtained with the medium solidified with agar, while in difcobactoagar only 1.4 of plantlets in replication was produced. This experiment suggested that agar and plant growth regulators in the medium were beneficial for producing embryo and plantlet from maize anthers.
4 Microarray Data Analysis for Detection and Classification of Viral Infection, Khadijeh Nazari, Ali Karami, Nezameddin Mahdavi Amiri, Fatemeh Pourali
DNA microarrays consist of collection of DNA microscopic spots that In order to form an array attached to a solid surface such as glass, plastic or silicon chip. The pieces of fixed DNA considered as a searcher. In this technology it is possible to test sample against thousands probes for specific genes. With this ability, arrays accelerate the biological investigations, gene finding, molecular detection and disease diagnosis. Microarray technology can be seen as a continued development of southern blotting. The most important stage in this technology is data analysis. To analysis such large data whit high degree of confidence and reliability needs reliable bioinformatics tools. Infectious diseases still is major problem for human. One of the most important application of microarray technology is the possibility of testing for the presence of thousands micro-organism in environmental and clinical samples only in a single excrement. Thereby we take an important step in rapid and accurate detection of infectious diseases. Here, we present E-Predict algorithm and DetectiV package that is based on species identification in microarray. We demonstrate the application of E-Predict and DetectiV for viral detection in a large publicly available dataset and show that DetectiV performs better than E-Predict. DetectiV is implemented as a package for R - powerful, open source software for statistical programming - that containing visualization, normalization and significance testing functions.
5 Enhancing iron oxidation efficiency by a native strain of Acidithiobacillus ferrooxidans via response surface methodology, and characterization of proteins involved in metal resistance by proteomic approach, Fahimeh Nemati, Daryoush Arabian, Rasool Khalilzadeh, Fazin Abbaspour Aghdam
The effects of different factors on growth and bio-oxidation efficiency of a native strain of Acidithiobacillus ferrooxidans have been evaluated by the utilization of response surface methodology, RSM. Medium pH and iron concentration were found to be the most significant factors while temperature and ammonia concentrations had the least weight within the ranges investigated. Optimum operational conditions for maximizing Fe2+ oxidation were found to be 31 oC, 7 g/Liron concentration, 4.5 g/Ltotal ammonium salt concentration and medium pH 1.85. Maximum recovery of 98% of Zinc is the main outcome of results as observed at 7 g/Lof Fe2+, under optimized experimental conditions. The response of a bacterial strain to metals toxicity also studied. The isolate showed good resistance to most of the toxic metals. The proteomics approach was used to identify the differentially expressed proteins under heavy metal stress. Four of the differentially expressed proteins were identified as major outer membrane protein of A. ferrooxidans, ribulose large bisphosphate carboxylase subunit, and holo synthase.
6 Determination of T-2 Mycotoxin Fusarium strains by HPLC with fluorescence detector , Reza Kachuei, Sassan Rezaie, Mohammad Hossein Yadegari, Naser Safaie, Abdol-Amir Allameh, Mohammad-Ali Aref-poor, Abbas-Ali Imani Fooladi, Majid Riazipour
T-2 toxin is the most poisonous trichothecene produced by Fusarium species especiallyF.sporotrichioides. T-2 toxin is a biological contaminant in a number of agricultural commodities that can cause severe diseases among humans and animals and even lead to death. The aim of the current study is the analysis of T-2 mycotoxin in Fusarium species by high-performance liquid chromatography (HPLC) combined with florescence detection and derivatization with 1-antroylnitrile (1-AN). Totally, 11Fusariumisolates and reference strains were studied. The isolates were tested for the T-2 toxin production after growing on rice substrate followed by using specific Multisep 225 Trich Clean up columns purification. In this study, T-2 toxin production was ranged from 197.05ug/kg to 8503.07ug/kg. This is the first study of T-2 toxin analysis by HPLC-F in Iran.
7 Determination of T-2 Mycotoxin Fusarium strains by HPLC with fluorescence detector , Reza Kachuei, Sassan Rezaie, Mohammad Hossein Yadegari, Naser Safaie, Abdol-Amir Allameh, Mohammad-Ali Aref-poor, Abbas-Ali Imani Fooladi, Majid Riazipour
T-2 toxin is the most poisonous trichothecene produced by Fusarium species especiallyF.sporotrichioides. T-2 toxin is a biological contaminant in a number of agricultural commodities that can cause severe diseases among humans and animals and even lead to death. The aim of the current study is the analysis of T-2 mycotoxin in Fusarium species by high-performance liquid chromatography (HPLC) combined with florescence detection and derivatization with 1-antroylnitrile (1-AN). Totally, 11Fusariumisolates and reference strains were studied. The isolates were tested for the T-2 toxin production after growing on rice substrate followed by using specific Multisep 225 Trich Clean up columns purification. In this study, T-2 toxin production was ranged from 197.05ug/kg to 8503.07ug/kg. This is the first study of T-2 toxin analysis by HPLC-F in Iran.
8 Biocontrol of Amaranthus retroflexus and Rumes crispus by NLP phytotoxine, a selective bioherbicide , Fatemeh Shakeri, Ali Mohammad Latifi, Morteza Mirzaei, Hamid Babavalian, Fatemeh Hashemlou
Non-beneficial and harmful weeds are plants that are unwanted, outside their home farms are growing and have the potential to exceed. This study was done in order to screening fungai and isolating NLP phytotoxine from them for selective biocontrol of Amaranthus retroflexus and Rumes crispus as a dicot, common and chemical herbicide resistance weeds. NLPs are effective just on dicot plants. Contaminated soil and dicotyledons plants were Collected from different regions of Iran. after collecting and culturing them, The effect of Supernatant from fungal cultures, was asseyd by spraying of 5 l /cm3 of it mid 20 l tween-20 on leaves of Amaranthus retroflexus, Rumes crispus and wheat as negative control that were cultured in MS media and pots in 3 replications with completely randomized design in laboratory and research green house of baqiatallah university. The effects were assessed according to numbering method. Finally, the QAT5and G7-1strains was selected from 9 top strains, because was more destructive than others on Amaranthus retroflexus and Rumes crispus respectively from necrosis to cell death with number 4 according to numbring method and has non-harmful effect on the wheat (Triticum aestivum). SDS-page results showed phytotoxine that was produced by QAT5 strain was a protein and this from G7-1 was non-protein. For better result on SDS-page protein was concentrated using by ammonium sulfat method, but about G7-1 again this outcomewas repeated. The protein purification of QAT5strain using FPLC showed the presence of a protein with about 24 kDa like other family members of this protein. Considering this fact that these phytotoxines according to the result had similarity features to what founded befor about NLPs, they are recommended as biocontrol factor of these weeds insteade of chemical herbicides.
9 Investigating the Effect of Dimercaprol on Phytoremediation of Lead by Zea mays,, Mohammad Amin Sharif, Ali Mohammad Latifi, Mohammad Ali Amani, Fatemeh Shakeri, Morteza Mirzaei, Parvin Noroozimoghadam
Application of plants to remove heavy metals from soil (phytoremediation) is expanding due to its cost-effectiveness as compared to conventional methods and it has shown a great potential. Since contaminants such as Pb have a limited bioavailability in the soil, methods to facilitate their translocation to the shoots and roots of plants are required for efficient phytoremediation. The present pots experiment were to investigate the effect of Dimercaprolchelator with different concentration (0, 1.5, 3 mmol Dimercaprolchelator kg-1). Also different ranges of Pb (100, 200 mg Pb kg-1) and a control group were investigated for the amount of Pb accumulation, by corn (Zea mays), sunflower (Helianthus annuus) and mustard (Sinapis arvensis). The results showed that the amount of Pb accumulation increased as the Pb concentration was increased. Also the results of this experiment showed that the addition of Dimercaprolchelatoris most likely to increase the bioavailability of Pb and consequently the accumulation of this heavy metal in the shoots. The highest accumulation of Pb was noticed with the highest does of Dimercaprolchelator (3 mmol Dimercaprolchelator kg-1) and Pb (200 mg Pb kg-1) in the shoots of Helianthus annuus.
10 Neuroprotective Effect of Coenzyme Q10 in Hippocampal Injury in Balb/c Mouse, Sara Rigi, Abolhassan Sakhae, Azadeh Niknam Asl, Akram Alizadeh, Fatemeh Beigi, Mehdi Mehdizadeh, Majid Ketabi, Mansooreh Soleimani
Coenzyme Q10 is a promising agent for neuroprotection in neurodegenerative diseases. Neuroprotective effects of Coenzyme Q10 demonstrated in some neurodegenerative diseases such as Parkinson, Alzheimer and etc. Hippocampus is home of these diseases. We assayed Coenzyme Q10 effects on Hippocampal injury model and our hypothesis is that Coenzyme Q10 has Neuroprotective effects in some neurodegenerative diseases via hippocampus. For this purpose 24 Balb/c mouse took in 4 groups: Control (Without any treatment), Vehicle (Treated with sesame oil as Coenzyme Q10 vehicle), Hyppocampal injury model (Treated with Trimethyltin chlorideneurotoxin, 2.5 mg per kg IP), and test (Treated with Coenzyme Q10 after Trimethyltin chloride injection, 10 mg per kg IP for 2 weeks). After two weeks brain harvested and hippocampus tissue assayed by Nissl and Tunnel staining. Hystological study showed significantly increase of normal cells and decrease of apoptotic cells in test group after Coenzyme Q10 treatment in hippocampus. This study showed Coenzyme Q10 has protective effects in hippocampus after injury and it seems that Neuroprotective effects of Coenzyme Q10 in some neurodegenerative diseases com from that.
11 Rapid Detection of Vibrio Cholerae by Polymerase Chain Reaction based on Nanotechnology, Gholam Reza Herfehdoost, Mahdi Kamali, Hamid Reza Javadi, Davood Zolfagary, Ali Choopani, Bahareh Ghasemi, Saeideh Hossaini
Quick identification of Vibrio Cholerae in epidemics is important, on the other hand; conventional methods are time-consuming and costly. The aim of this study was to develop a rapid, inexpensive and high sensitivity method for quick identification of Vibrio Cholerae. For this purpose we designed a PCR detection based on magnetic nanoparticles for identification of bacterial DNA by PCR Dynabead. So we used the biotinylated Probe for binding to DNA extracted from Vibrio Cholerae and other bacterial species (Salmonella, Shigella, Pseudomonas, E.coli) Using magnetic bead isolated with magnetic field, the Vibrio genome-specific primers (HlyA) for pathogen detection (PCR) was used. The results showed specific band was just for Vibrio Cholera (PCR positive); therefore designed probe was specific for Vibrio Cholerae. According to the findings, this study is characterized the high sensitivity of PCR using biotin-containing probes for DNA of Vibrio Cholera in contrast to the traditional methods.
12 The Inhibition Effects of Two Species of Astragalus Extracts on Mushroom, Azadeh Taherian, Alireza Jahangiri, Forogh Namjoyan, Seyed Reza Hajhosseini
Melanin pigment plays a critical role in camouflage and protection against harmful effects of solar radiation. Melanogenesis is under complex regulatory control by multiple agents. Tyrosinase is a multifunctional, which catalyzes the first two steps in mammalian melanogenesis. In this study, inhibitory effects of Astragalus fasciculifolius and Astragalus gypsicolus on diphenolase activity of mushroom tyrosinase were evaluated. .Ethanol 80% extracts were prepared for screening tests. The IC50, Ki, Km and Vm values of Astragalus fasciculifolius and Astragalus gypsicolus were measured then compared them. We found that Both extracts show mixed type inhibition on mushroom tyrosinase when L-DOPA was used as a substrate.
13 Production and Purification of Polyclonal Antibodies against Diphtheria Toxin, Mohammad Ali Arefpour Torabi, Gholam Reza Olad, Shahram Nazarian, Jafar Salimian, Samaneh Khodi, Mohamad Javad Bagheripour
Diphtheria is a fatal disease caused by exotoxin of Corynebacterium diphtheria. This toxin consists of two chains, catalytic chain (A) and binding (B) chain. By binding chain (B), the toxin binds to its receptor on numerous body cells such as myocardial, kidney and peripheral nerve cells. After entering, catalytic chain (A) inhibits protein synthesis and finally can cause cell death. At this time, the toxoid form of diphtheria toxin is used as vaccine. The aim of this study was the immunological analysis of the mutated synthetic catalytic subunit of diphtheria toxin in laboratory animals as a vaccine candidate, in addition to polyclonal antibody production and purification against diphtheria toxin. For this purpose the Dtx recombinant protein (with two mutant: A158G and G52E) was expressed using pET28a/DtxA plasmid in E. coli Bl21DE3 host. Then, recombinant protein, as a candidate vaccine, was extracted and purified. After evaluating and confirming the protein by SDS-PAGE and western blotting, immunization carried out in laboratory animals. Finally, followed by antibody titration by ELISA, antibody purification performed as well.The mutated recombinant protein prepared from an optimized expression was extracted and purified. Then, this protein was confirmed by SDS-PAGE and western blotting. ELISA results showed a satisfactory immunization of animals by this protein. Polyclonal antibody production and purification against diphtheria toxin was performed by G protein column and confirmed by ELISA. ELISA results showed a high titer of polyclonal antibody against diphtheria toxin in animal's serum after immunization by recombinant DTx protein.
14 Synthesis of Zinc Oxide and Chromium (III) Oxide Nanoparticles with Diverse, Ramin Karimian, Farideh Piri, Seyed Javad Davarpanah
The use of an inorganic phase in water-in-oil (w/o) microemulsion has received considerable attention recently for preparing metal oxide nanoparticles. This is a technique, which allows preparation of ultrafine metal oxide nanoparticles within the size range 40 to 80 nm. Preparation of nano ZnO and Cr2O3 studied, investigated in the inverse microemulsion system. Therefore the nucleation of metal particles proceeds in the water capsules of the microemulsion. Zinc oxide and Chromium (III) oxide nanoparticles powder has traditionally been used as a pigment and diverse physiological properties. Physiologically important nanoparticles are currently under investigation for their bio-medical applications as well as for therapeutics.
15 Micropropagation of Common Yew Using Embryo Culture, Seyed Javad Davarpanah, Mehrdad Lahouti, Ramin Karimian
Common yew Taxus baccata L. is an evergreen, slow growing tree, indigenous of hyrcanian forests in Iran. Discovery of Paclitaxel as an anti tumor agent in the bark of Taxus has increased importance of this plant for treatment of breast and ovary cancer, especially 7000 women die annually due to breast cancer in Iran. Regarding destructive method of Paclitaxel production and protection of the yew as an endangered tree in Iran, micropropagation using embryo culture can be considered as an efficient method to increase Taxus resources in Iran. In this regard, effects of different culture medium compositions, active charcoal concentrations and gibberellic acid on embryo culture of Taxus baccata L. were studied. Murashig and Skoog culture medium at half and complete strengths of macronutrients and active charcoal at concentrations of 0, 2, 4 and 6 g/L were used. Based on experimental data MS/2 + 2 g/L active charcoal appeared to be the most effective treatment for successful embryo culture.
16 Study of Paraoxonase -1 Gene Polymorphism in a Healthy Population of, Kobra Chehari, Farideh Sepahvand, Sirous Ghobadi, Ahmad Ismaili, Ezat Rafiei Alavy
Human serum paraoxonase (HuPON1: EC 3.1.8.1), a calcium-dependent esterase, is synthesized in the liver and widely distributed in tissues including liver, kidney, intestine, and serum, where it is associated exclusively with high-density lipoprotein. Human paraoxonase-1 plays an important role in prevention of atherosclerosis and also protection against organophosphate-induced neurotoxicity. Paraoxonase-1 shows 2 common polymorphisms: Q/R at position 192 and M/L at position 55. In this study, paraoxonase-1 192 and 55 polymorphisms were investigated in 64 healthy Iranian individuals. Genomic DNA was isolated from whole blood by the Bartlett method, and paraoxonase-1 genotypes were determined by polymerase chain reaction amplification followed by restriction isotyping and gel electrophoresis. The chi-square test was used to evaluate the Hardy-Weinberg equilibrium. The genotype frequencies for paraoxonase 1-Q192R were approximately 47% (QQ), 41% (QR) and 12% (RR) and for paraoxonase-1 M55L, 44% (LL), 44% (ML) and 12% (MM). Thus, the frequency of alleles R, L, Q, and M were 0.33, 0.66, 0.67, and 0.34 respectively. In conclusion, the frequencies of paraoxonase-1 192 and 55 polymorphisms in this group of Iranian population were different from those seen in other Asian populations from Japan and China but similar to European (Caucasians).
17 Investigation of the Effect of Substrate Conditions on Electron Transfer of Glucose Oxidase, Khadijeh Eskandari, Sharareh Sajjadi, Amir-Homayon Keihan, Mehdi Kamali, Jamal Rashidiani, Zahra safiri
Direct electron transferring of glucose oxidase was investigated on reduced graphene and graphene oxide templates. The direct electrochemistry glucose oxidase on graphene showed a cyclic voltammograms corresponding to the FAD/FADH2 redox couple with an anodic, cathodic and formal potential of -430, -460 and -445 mV, respectively in 0.1 M phosphate buffer solution and air saturated condition for similarity of in vivo usage. The cyclic voltammograms of glucose oxidase on graphene is reversible. Also, the voltammograms results show, the current intensity of glucose oxidase on graphene is high, due to fast electron transferring. Moreover, the linear rang concentration of glucose on graphene are 0.49 M. These studies make useful insight into the enzyme immobilization on nanoparticles for biosensors and bio-fuel cell preparation.
18 Growth factor Containing Hydrogels for Tissue Engineering Applications, Hamid Babavalian, Ali Mohammad latifi, Mohammad Majid Sepantafar, Hossein Mohammadi, Fatemeh Shakeri, Smaneh Khodi
Regenerative medicine provides substitutes for organ transplantation, which is restricted in applicability due to immune responses against allograft and the large discrepancy between the need for organs and the number of available transplantation. An artificial environment is allowing cells to induce tissue regeneration. Growth factors (GFs) play a significant role in the cell fates in their microenvironment. Nevertheless, the short half-lives of GFs and poor in vivo stability infiltration suggest that the classical routes are useless and insist on use of a drug delivery system (DDS). In this review paper, growth factor-based tissue regeneration using polymeric hydrogels is reviewed to show great potential ability of hydrogels in tissue engineering (TE).
19 Optimization of Callus Induction in Pennyroyal (Mentha pulegium), Elaheh Darvishi, Elham Kazemi, Danial Kahrizi, Sayed Reza Chaghakaboudi, Yavar Khani
Findings in relation to the inhibitory effects of natural substances against microorganisms showed that herbal products may be alternatives to synthetic and chemical drugs due to their significant therapeutic effects. The pennyroyal (Mentha pulegium) as an important medicinal plant, belongs to the Lamiaceae family is applied in the pharmaceutical and food industries. In recent years, plant tissue culture techniques have appeared as a powerful tool for the micropropagation and breeding of many plant species. The aim of current study was to find the best medium composition for callus induction in M. pulegium. For this purpose, three explants (leaf, root and stem) of M. pulegium were cultured on MS medium supplemented with BAP (0.5 and 1.0 mg/L) and 2,4-D (0.0, 1.0, 2.0 and 4.0 mg/L) in a factorial experiment. The results showed that the leaf explant had the highest effect on callus induction (9.65 mm in callus diameters at 28 days after culture) and hormone levels of 1 mg/L 2,4-D and BAP-free identified as the most efficient concentrations for callus growth rate (0.29 mm/d). Also statistical analysis demonstrated that among interaction effects t he leaf explant in 1 mg/l 2,4-D and BAP-free found as the highest effect for callus induction (12.35 mm in callus diameter at 28 days after culture).
20 In Silico Analysis of Amino acid Substitutions in DNA gyrase subunit A, Payam Behzadi, Elham Behzadi, Mehrdad Moosazadeh Moghaddam, Ali Najafi, Reza Ranjbar
The broad consumption of antibiotics such as fluoroquinolones and the genetic adaption of opportunistic pathogenic bacteria including Pseudomonas aeruginosa, has led to develop of floroquinolone-resistant strains of Pseudomonas aeruginosa. Thus, the aim of the present study is to reveal the types of hot spot mutations occurring in selected DNA gyrase subunit A gene in fluoroquinolone-resistant Pseudomonas aeruginosa strains. For this purpose, 70 fluoroquinolone-resistant partial cds genes of Pseudomonas aeruginosa TOHO strains (P12-P377) for DNA gyrase subunit A have been aligned with wild genes for DNA gyrase subunit A by BLAST program and multiple alignment, to detect probable gyrA mutations. Based on analyses hot spot mutations including Thr-83?Ile (92.44%), Ile-83?Thr (1.33%), Asn-87?Asp (0.44%) existing as single and Thr-83?Ile & Asn-87?Asp (5.78%) as double substitutions were detected. According to this survey, the point mutations in DNA gyrase subunit A gene need more consideration about fluroquinolone-resistant Pseudomonas aeruginosa isolates. The strains with hot spot SNP mutations may rise a huge concern from epidemiologic aspect for antibiotic therapy treatment.
21 Genotype-Dependent Differential Response of Cereal Germinating Seeds Against, Smaiel Malek Hosseini, Milad Ahmadi Najafabadi, Hossein Askari, Abbas Saidi, Ehsan Sadegh Nejad
Land mines are one of the tragic problems to human security in many countries. Among many techniques suggested detecting landmines, plant-based detectors can be used as a reliable tool for unexploded ordnance detection. This study was conducted to determine different concentrations effect of trinitrotoluene (0, 5, 10, 15, 20, 30, 45 and 60 ppm) on germination and early seedling growth of 6 species of cereals including Barley, Wheat, Durum wheat, Oat, Rye and Triticale. In contrast to oat and durum, barley and wheat genotypes had the best germination and germination index against different trinitrotoluene (TNT) concentrations. TNT showed no impact on the alpha amylase activity of the tolerant (Barley var. Bahman) and susceptible (Durum wheat) genotypes. Under higher TNT concentration, barley had the highest root length and root number and also was the best in dry matter allocation towards root tissues. We measured the different parameters for roots of 6 species of cereals, that oat was sensitive plant when exposed to TNT than others. With emphasis on the wide-range adaptation of cereals to climate and soil characteristics and their extensive and fibrous root systems, it is suggested that barley genotypes especially Bahman variety can be a confident plant to carry the essential genetic structure for explosive detection.
22 Optimizing Transfection of Umbilical Cord Mesenchymal Stem Cells Utilizing, Nasibeh Daneshvar, Peyman Ghoraishizadeh, Mohadeseh Hashemboroojerdi, Shraddha Raikar, Parvaneh Mehrbod
Lipofectamine LTX is a cationic based transfection reagent that offers high levels of different transgenes expression in a huge number of cell types in vitro utilizing an easy and rapid protocol. The level of transgenes expression depend on some experimental variables containing cell density, lipofectamine and DNA concentrations, lipofectamine -DNA complexing time, and the presence or absence of medium at the transfection time and antibiotics in medium. The significance of these factors in lipofectamine LTX based transfection will be considered. The main objective of this study is optimizing different situations for transfection of umbilical cord mesenchymal stem cells (UC-MSCs) with lipofectamine LTX. For this purpose, UC-MSCs were transfected with lipofectamine LTX utilizing minicircle plasmid containing green fluorescent protein reporter gene. In this study, the medium existence on cell transfection efficiency as well as effects of time for culture, the ratio of lipofectamine LTX to plasmid DNA, the repetition times of transfection and volume of medium on transfection efficacy were evaluated, and the transfection efficacies were also compared. The growth rate of MSCs was associated with the density of the cells. The medium changing 4 hours after transfection certified the normal cells growth. Nevertheless, the freshness and amount of medium demonstrated no substantial effect on the cell state. The 1:2 ratio of plasmid to ipofectamine LTX (3ng:6l), the transfection efficacy was pleasing. The transfection cells with different ratios of plasmid to lipofectamine LTX displayed substantial differences18 h post transfection. The differentconditions of MSCs transfection with lipofectamine LTX was optimized, which presented a reference for various adherent cells transfection.
23 Synthesis and Characterization of Co3O4 Nanotubes to Prepare Variety of Electrochemical Biosensors , Seyed Javad Davarpanah, Ramin Karimian, Farideh Piri
Cobalt (II) Chloride was used as a precursor to prepare Co3O4 nanotubes in reverse micelles. Tween 80 was added as surfactant. The products were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM) and transmission electron microscopy. The diameter of Co3O4 nanotubes was about 58 nm and the average length of them was around 261 nm. The study provides a simple and efficient route to synthesize Co3O4 nanotubes at room temperature. Tremendous demands for electrochemical biosensors with high sensitivity and reliability, fast response and excellent selectivity have stimulated intensive research on developing versatile materials with ultrahigh electrocatalytic activity.
24 A Decrease of Reelin Expression in Neonatal Ventral Hippocampal Lesion Model, Fereshteh Mozaffarian, Shima Ababzadeh, Akram Alizadeh, Esmaeel Sharifi, Majid Katebi, Farzaneh Mohammadzadeh, Sara Soleimani Asl, Pegah Zoalfaghar, Mansooreh Soleimani
Schizophrenia affects 1% of population. Neonatal ventral hippocampus lesion (NVHL) model of schizophrenia designed in 1993 by Lipska and is a widely studied developmental animal model of schizophrenia. NVHL rats mimic many of the symptoms of schizophrenia in detail. We studied this model in molecular level and reelin expression in it. Reelin is an extracellular matrix glycoprotein that regulates some processes in CNS development and reduces significantly in schizophrenia. For this study, animals (male pups) take into 3 groups: control, sham and experiment. The lesion made by injection of 0.3 l. Isotonic Neurotoxin withstereotaxic surgery in age 7 day and body weight 11-15 gr. Social behavioral and stereotypic movement assessed in age 56 day then reelin expression in frontal cortex evaluated by western blotting. Behavioral analysis and histological studies demonstrated the schizophrenia model. Western blotting of reelin protein in frontal cortex and hippocampus show a decrease of reelin (P value: 0.012) in experimental group as compared to control and sham group. So, in the NVHL as a common and more similar model of schizophrenia reelin expression significantly decreases in frontal cortex and hippocampus that means this model in molecular pathways is similar to the disease.
25 Nanoparticles in Vaccine Development, Abbas Hajizade, Firouz Ebrahimi, Ali-Hatef Salmanian, Ayyoob Arpanae, Jafar Amani
Vaccination has greatly improved human health. Despite of all improvements in this field, there is not an efficient vaccine for many diseases, and of the available ones, some could not produce a long-term immunity. Recently, there have been many researches on the applicability of nanostructures as an efficient system for vaccine delivery, and the initial results have been promising. Their potential adjuvanticity, capability of the stimulation of both humoral and cellular immunity responses, more stability in environmental conditions, possible targeted vaccine delivery, the need for low quantity of proteins (in the case of subunit vaccines), etc., are of the main reasons that this area has gained many interests. Here, we try to review the main nanostructures that could be act as a delivery vehicle in vaccine delivery
26 Identification and Distribution of Anaerobic Responsive Elements (AREs) in Genes Functional Categorization of Arabidopsis thaliana, Ehsan Sadeghnezhad, Hossein Askari, Sattar Soltan, Fatemeh Honarvar
The evolution of biota had been tightly linked with formation of atmospheric di-oxygen (O2).Since the shift from anaerobic to aerobic life results in reprogramming gene expression through the regulatory interactions between transcription factors and their target sequences, the anaerobic responsive elements (AREs). The purpose of this study is to consider AREs; the most frequent and divergent motif in comparison to eighty four plant cis-regulatory elements, in order to clarify their biological function in Arabidopsis thaliana. A genome wide survey revealed the frequency, occurrence number and positional state of each anaerobic element in Arabidopsis genome. The evidence was obtained that among all AREs, TTCCCTGTT, (A/C/G)AAAAACAAA and AGCAGC in silico discovered elements, are noteworthy responsive elements in Arabidopsis. In addition to, our data suggest that the evolutionary process of anaerobic responsive elements may have been directed toward gradual decrease in occurrence numbers of AREs and gradual increase of their ineffectiveness in Arabidopsis genome. We also classified Arabidopsis genes according to existence of ARE elements in promoter regions of them to seven main groups including expression, primary metabolism, secondary metabolism, signaling, growth and development, stress and misc.
27 In Silico Design of a Multimeric Polytope as a Highly Immunogenic DNA Vaccine Against Human Cytomegalovirus, Elaheh Sabbaghian, Fatemeh Roodbari, Alireza Rafiei, Jafar Amani
Human cytomegalovirus is one of the most common pathogenic viruses all over the world. In congenital infection leads to neurologic severe disorders and even death of fetus and in individuals with immunosuppression may also cause severe clinical symptoms. Multiple evidence indicate that among several strategies, epitope-based vaccine (EVs) that can induce both humoral and cellular immunity responses, are the most important and have numerous potential profits. In this study, we select the viral surface glycoprotein B and phosphoprotein 65 and 150 with the highest antigenic and immunogenic properties, that have the most important role in induce cellular and humoral immune responses. Bioinformatics tools, as a standard and developed approaches use for epitope mapping. Epitope discovery greatly accelerate by in silico prediction methods with in vitro and in vivo verification. Bioinformatics methods and epitopes identification algorithms were used in order to selection of cytomegalovirus immunodominant epitopes, detection of each epitope antigenicity and design chimeric gene construct. The chimeric protein showed high antigenicity in vaxiJen analysis. Also further immunoinformatic analyses in order to predict the discontinuous and continuous B and T cell epitopes and MHC binding peptides affinity were used. The study results show that protein structures were suitable. Therefore it can be expected that construct is proper subject for practical experiments and stimulus for humoral and cellular immune responses.
28 Improvement of Thermal Stability of DFPase by In silico Methods, Morteza Mirzaei, Ali Mohammad Latifi, Rahim Jafari
Efficiency of enzymes which are used in industrial or environmental applications is highly dependent on their thermal stability. In this study, the stability of DFPase has been evaluated after introducing disulfide bonds to the structure. The results obtained from a series of protein design software were subjected to molecular dynamics simulation at different temperature to test the performance of such combinatorial procedure. Amount several designs, mutation M5 showed desirable thermostability via molecular dynamics simulation and normal mode analysis. As it clearly depicted, such in silico structural investigations would be resulted in reducing the numerous choices of experimental options as it was undergone a series of computational evaluation previously.
29 Biodecolorization of Textile Effluents by Autochthonous Fungi, Maryam Mazaheri Tehrani, Mahnaz Mazaheri Assadi, Hamid Rashedi
Textile effluents contain different kinds of chemical dyes and many mutagenic and carcinogenic substances. Discharge of these effluents into terrestrial and aquatic ecosystems causes harmful effects. In this research, decolorization of a textile effluent by nineteen different strains of fungi was investigated. Removal activity of dyes was in order of Aspergillus > Rhizopus > Fuzarium > Penicillium > Saccharomyces. The selected fungal strain was capable of decolorizing textile effluent at original pH of effluent (8.5- 9) by adding 0.6 and 0.2 g/L of sucrose and NH4Cl as additional carbon and nitrogen sources respectively at 30?C. 98 % of dyes were absorbed from effluent during the exponential growth phase in the presence of a biodegradable substrate such as sucrose under shake flask conditions. Dyes strongly bound to the fungal biomass required extraction with methanol for their removal. The dyes pigment yellow (PY 74) and reactive blue (RB 38) were totally decolorized after 6 days at initial concentration of 250 ppm by terrestrial Aspergillus niger.
30 Effects of Different Concentrations of 2, 4-D and Kinetin on Callogenesis of Taxus Brevifolia Nutt, Ramin Karimian, Mehrdad Lahouti, Seyed Javad Davarpanah
Taxus is an endangered plant which is the only commercial source of paclitaxel, which is used for treatment of ovary and breast cancer. As production of this valuable drug is a destructive process, other alternatives should be considered for its sustainable production. Plant tissue and cell culture is a promising method for production of secondary metabolites. In order to optimize yew callus culture effects of 16 combinations of two plant growth regulators, 2, 4 dichlorophenoxyacetic acid (0, 0.5, 1 and 1.5 mg/L) and Kinetin (0, 0.1, 0.5 and 1 mg/L) on callogenesis of Taxus brevifolia twig explants were studied. Considering growth criteria, fresh weight (33.5 mg), dry weight (3.88 mg) and callogenesis ratio (1.45), based on statistical analysis the best plant growth regulators combination for induction and growth of callus of twig explants was appeared to be 1.5 mg/L of 2, 4- dichlorophenoxyacetic acid and 0.1 mg/L of Kinetin.
31 Simultaneous determination of Zinc and Lead in blood samples by Differential Pulse Polarography, Fatemeh Abolhasani, Mohammad Ali Amani, Hossein Aghamollaei, Morteza Mirzaei, Ali Mohammad Latifi
In present study we determine the Zinc and Lead levels in human blood samples by differential pulse polarography under optimum experimental conditions (pH=7.5, Scan rate=5mV/S and pulse amplitude=50mV) the polarographic reduction peaks of Zn and Pb were examinedin the -0.72 V and-0.15 respectively. The zinc and lead content can be determined by in comparison with their standard solutions. According to the obtained results differential pulse polarography is more reliable, rapid and sensitive method for determination of zinc and lead levels in the blood than atomic absorption spectroscopy.
32 DNA Electrochemical Nanobiosensors for the Detection of Biological Agents, Mohammad Ebrahim Minaei, Mojtaba Saadati, Mostafa Najafi, Hossein Honari
In this study, we explained a nanobiosensor for DNA sequence detection, featuring sequence specificity, cost efficiency, speed, and ease of use. Without the need for labels or indicators, it may be ideal for the detection of biological agents. This review describes recent advances in electrochemical impedance spectroscopy (EIS) with an emphasis on using nanoparticles and nanotechnology. A powerful biosensor system requires a high-performance biosensor component as well as a user-friendly instrumental setup. However, biosensor setups have to be adapted to specific applications. Rapid, selective and sensitive detection technologies for biological agents are critical in clinical diagnosis, environmental monitoring and food safety. Recent developments in nanomaterial create many opportunities to advance DNA sensing and gene detection. The fact that gold nanoparticles are able to provide a stable immobilization of biomolecules that retain their bioactivity is a major advantage for the preparation of biosensors. Although, there are a lot of researches reporting electrode modification by different nanomaterial to improve the DNA biosensor performance, the preparation of nanomaterial or the electrode modification strategy is often relatively complex. Furthermore, some DNA biosensors based on nanomaterial modification are still very limited for the improvement of DNA biosensor performance. Thus, the construction of nanostructure modified electrode by a simple strategy to improve the DNA detection sensitivity is highly desirable.
33 Different Methods for Diagnosis of Brucella and Legionella spp. in Various Samples, Zahra khazaei, Nadia Jahandeh, Fatemeh Saberi, Mehran Fesahat, Reza Mirnejad
Community-acquired pneumonia and brucellosis are of the main agents causing mortality in the world. The most important limitations for the identification of these agents include their high resistance against environmental conditions, impossibility of rapid and on time diagnosis, low level of infective dose, and lack of existence of vaccine against many types of them. Hence, on time action and diagnosis of agent of infectious diseases has been regarded as a solution for preventing incidence of this type of disease. Because of the danger of Brucella and Legionella bacteria, studying isolation methods is one of the most important measures regarding detection of agent of this type of attacks. The present study is a review study, which is conducted using existing studies and also using library method in order to investigate the detection of two mentioned bacteria in various samples.
34 A Study on the Mechanism of Aggregation of Therapeutic Reteplase Protein by Using the Monomer-Loss Model , Soheyla Ershadi, Hamid Rashedi, Ahmad Fazeli
Aggregation of pharmaceutical proteins reduces the efficiency and increases the cost of production. It can also lead to the reduced efficacy of drug or cause side effects on the patients body. Investigating how to create them plays an important role to find agents that prevents the aggregation. This study was allocated for understanding the mechanism of formation of the reteplase protein using thermal stimulation. Aggregation was studied by ultraviolet spectrometry, and observation at 4, 25, 50 and 70C, the concentration of protein monomer was measured by using a spectrum of 360 nm and 280 nm. At 4C, there was no significant change in monomer concentration for a month. By increasing the temperature to 25?C, aggregation process was slow, but at 70C, the reaction was carried out at a rapid rate less than 2 hours. In order to investigate the mechanism of reteplase aggregation, some kinetics that was presented in of monomer-loss models were used. Experimental data was fitted in three pre balance core, self-catalytic and slow start models using MATLAB. The best fit was obtained using optimization methods. Best fit for self-catalytic model is (R2> 0.98). For other two models (R2
35 Utilizing Different Supports and Comparing their Performances in the Construction of Morphine Rapid Detection System Based on Lateral Flow Assay, Housna Shamloo Ahmadi, Mohammad Heiat, Hamid Rashedi, Ali Mohammad Latifi
Lateral flow assay, a simple and rapid test to detect various agents; have found many applications in different fields. One of the most important issues in the construction of a Lateral Flow assay is the increasing of its sensitivity and efficiency by processing of analytical layer. This study, aimed to investigate the performance of, Nellulose, Nitrocellulose, Nylon and PVD as different analytical layer in morphine Lateral flow strip as a model. Cyanogens bromide, acetonitrile, methanol and specific polymers were used to activate cellulose, nitrocellulose, PVDF and nylon respectively. BSA-morphine and anti morphine polyclonal antibodies were immobilized orderly in test and control bands different concentrations of morphine were prepared as sample solution. Results showed that activated supports have better detection level in comparison with control strips. Among activated supports nitrocellulose showed more reliable results rather than others and introduced as a suitable support for construction of analytical layer and antibody immobilization. Nylon and PVDF supports due to its hydrophobic nature and cellulose due to high capillary property and non-uniform texture showed inappropriate results. Using proper supports for design of lateral flow strips can improve the sensitivity and detection level of this system.
36 The Relationship between the Presence of Enterotoxin Type B Gene and Antibiotic Resistance in Staphylococcus aureus, Ali Choopani, Mohammad Heiat, Elham Amini, Mojtaba golpuch, Hossein Aghamollaei
Staphylococcus aureus is a hazard to human health since they can cause a wide variety of hospital-associated infections ranging from minor skin infections to post-operative wound infections and food poisoning which produces many different virulence factors, including enterotoxins (SEs). Although studies have been done regarding the difference between virulence factors of sensitive strains and resistant to antibiotics, the aim of this study was to investigate this topic in different patients. This cross-sectional study was performed on 100 patients admitted to a hospital in Tehran. After preparing of wounds samples, antibiogram study was done by disc diffusion method and prevalence of staphylococcal enterotoxin type B or seb gene was confirmed by Polymerase Chain Reaction. Data was analyzed using SPSS 17 software. Results showed that the highest percentage of isolates with positive seb gene is about 7% which related to amoxicillin (6.6%), penicilline and cotrimoxazole (6.5%). More than 90% of isolate are resistant to amoxicillin, penicillin and cotrimoxazol. According to results, a significant relationship between seb gene and resistance to related antibiotics was not observed.
37 Chemically Synthesis and Characterization of Iron and Nickel Oxide Nanoparticles for Biomedicine, Seyed Javad Davarpanah, Ramin Karimian, Farideh Piri
Magnetite (Fe3O4 and NiO) nanoparticles with a size range of 4060 nm were prepared by sol-gel technique in nano and micro reverse micelles (water- in-oil). The surface properties, size, morphology and crystallographic structure of Fe3O4 and NiO particles are characterized by means of X-ray diffraction, transmission electron microscope and scanning electron microscope which will give much valuable information about these materials. In addition, synthesis of nanoparticles can be easily implemented because it is simple and environmentally friendly.
38 Analysis of Antibiotic Resistance and Antimicrobial Effects of Enterococcus faecium and Lactococcus lactis Isolated from Khorramabad Traditional Cheeses, Sadegh Rezapour, Enayat Ghahremani, Mahnaz Mardani
Enterococcus and lactococcus are Gram-positive cocci that often occur in pairs (diplococci) or short chains, and are difficult to differentiate from streptococci on physical characteristics alone. Enterococcus faecium because have concern antibiotic resistant, consider difficult as probiotic. The results of an assay show that a probiotic E. faecium strain might be a potential recipient of Vancomycin resistance genes. For analysis of this concept, 13 samples of traditional cheeses were collected from different areas in Khorramabad city and identified with using phenotypic methods, and then the bacteriocin was extract from indentified bacteria. Agar diffusion method was used to assay the antimicrobial activities of bacteriocin produced by isolated bacteria against Pseudomonas aeruginosa, Proteus vulgaris, Staphylococcus aureus, E. coli, Bacillus cereus and Bacillus subtilis. On the other hand, antibiotic resistance of these bacteria was test using antibiogram method. The results showed that some of bacteria such as P. aeruginosa, S. aureus, and E. coli are intermediately resistance while P. vulgaris was completely sensitive and B. cereus and B. subtilis were resistant. Also, the Enterococcus faecium was resistant to kanamycin and trimethoprim antibiotics and intermediatly to clindamycin and tetracycline, and sensitive to amoxicillin and erythromycin. Lactococcus lactis was sensitive to trimethoprin, amoxicillin, tetracycline, and erythromycin while was resistant to kanamycin and clindamycin. In this study both bacteria enterococcus faecium and lactococcus lactis had an inhibitory effect on pathogenic bacteria and, these bacteria also have an appropriate antibiotic resistance to most antibiotics.