NEW CELLULAR & MOLECULAR BIOTECHNOLOGY JOURNAL
Year:2016 | Volume:6 | Issue:23|Papers Count:17

DNA catalysts (Deoxyribozymes, DNA enzymes; DNAzymes) are catalytic artificial single-stranded DNA molecules. Using the technique of in vitro selection, individual DNA catalysts have been identified that catalyze RNA cleavage, RNA ligation and a wide range of other chemical reactions. DNA catalysts have been used in vitro as biochemical tools, analytical tools and sensors. DNA catalysts have also been utilized as in vivo therapeutic agents to target specific mRNA. Although many conceptual and practical challenges remain to be addressed, DNA catalysts make new promise for applications both in vitro and in vivo. In this review, we are focusing on definition and different types of DNA catalysts, in Vitro Selection of DNA catalysts and their therapeutic applications. This review also describes the challenges and approaches to overcome obstacles involved in using DNA catalysts in the treatment of different diseases including cancers, viral and bacterial infection.

Aim and Background: Cytokeratins (CKs) constitute the largest intermediate filamentous protein that during the development of malignancy, the original CK profile of the cell is frequently variable. Cytokeratin 19 (CK19) is known as an epithelial cell marker and CK19 expression was seen in more than 90% of breast cancers. In this study, CK19 mRNA expression by the real-time PCR (Polymerase Chain Reaction) in luminal A cell lines was assessed. Materials and Methods: MCF7 and T47D breast cancer cell lines were purchased and cultured according to the recommended protocols. The cells were collected, counted and RNA was extracted from cell lines. The mRNA of CK19 as the target gene and GAPDH (Glyceraldehyde 3-phosphate Dehydrogenase) as a reference gene by real-time PCR was amplified. Quantitative CK19 expression was calculated using the DDCt method. The average ΔCt of CK19 and GAPDH was calculated in each cell line as well as the DCt of GAPDH as a calibrator in different cell lines was quantitatively reported.Results: This method can be used for detection of CK19 tumor marker expression, in the cells. On the other hand, the same expression in the MCF7 and T47D breast cancer cell lines was shown. Conclusions: Expression of CK19 is the same, in the luminal A breast cancer cell lines. Molecular markers could be used for cell Lines classification that it is significant in breast cancer research and it is helpful in detection and treatment of the disease.

Introduction and purpose: Various factors, such as sexually transmitted diseases, spinal cord injuries and radiotherapy after catching various cancers, which is the result of insufficient or no production Spermatozoa, leading to infertility in men. Today, by creating of therapeutic cell technology with mesenchymal stem cells, it is possible to treat many diseases. Meanwhile, mesenchymal stem cells of human umbilical cord Wharton's Jelly, have been considered as a resource that is frequently available and not rejected by the immune system. The first step in using of these cells is proving that they are fundamental. Therefore, in this study molecular stemness these cells was investigated with.evaluate genes expression of CD34, CD44, CD105 Method: Stem cells from the umbilical cord Wharton's jelly were isolated and cultured, total RNA was extracted from the third passage took place after the synthesis of cDNA, gene expression was determined by RT-PCR. The mesenchymal cells are exposed to different doses of BMP4 and medium terms obtained from newborn mice testicular cells and bed Nano fiber PLLA, To their differentiation. into the germ-like cells be Examined Results: The results showed that these cells have considerable proliferative potential and gene CD105, CD44 is expressed before the differentiation, but the expression of CD34, a marker of hematopoietic cells that is not seen. Putting the cells exposed to the distinction medium after the third. passage, leads them to the germ-like cells Discussion: Laboratory culture and enrich the germ cells derived from other sources such as umbilical cord mesenchymal cells is a practical method for the initial research and development germ.cells. Umbilical cord stem cells expressed genes are a strategy can be used to treat infertility.

Aim and Background: Nowadays in medical fields, the production of enzymes that are closely associated with human health has a special position in medical research. One of the environmental problems affecting human health is environmental pollution of which organophosphorus compounds (OPs) are notable. Therefore, the detoxification of OPs is important for human health. In this field, enzymes that can degrade OPs have been identified such as OPH. OPH production with a high performance is one of the goals of protein engineering. So the use of techniques applied to measure the enzyme activity is important. Several methods by using various Ops substrates can be designed. Material and Methods: In this study, we have used ISE to assay the degradation of DFP and spectroscopy and FPLC to assay the degradation of Diazinon and Chlorpyrifos. Then, the results were compared.Results: The results showed that ISE is a good method in case of DFP substrate due to the production of F- ion in presence of DFP. In the case of Chlorpyrifos and diazinon spectroscopy is more appropriate due to the changes in optical absorption of the reaction products. The FPLC method is an applicable method for all substrates Conclusion: It is possible to use various substrates at different wavelengths along with a remarkable significant difference in absorption and concentration by FPLC, therefore, it is suggested as a highly sensitive method for enzyme assay.

Aim and Background: To study the effect of irrigation and zinc nutrition on yield and essential oil production of (Menta piperita L.), a field experiment was conducted at College of Agriculture, Islamic Azad University of Khoy during spring of 2013. Material and methods: The experiment was conducted as a factorial based on complete randomized block design with four replications. The effects of irrigation in four levels including 100, 80, 60, 40 % (FC) and zinc nutrition in four levels including 0 (control), 2.5, 5, 10 mg/kg (ZnSO4) were investigated.Results: The results showed that the effect of irrigation on leaf length, leaf width, leaf number, stem diameter, number of internodes, number of branches per plant, plant height, fresh weight and shoot dry weight, leaf fresh weight and leaf dry weight were significantly (p £1%). The results also showed that zinc spraying on leaf length, number of leaves, stem diameter, leaf dry weight, total plant weight and percentage of essential oil was significant (p£0.01). The highest oil yield (20.81 kg/ha) were obtained with 100% humidity. Zinc spraying on the leaf width, number of branches, plant height and dry weight was significant (p£0.05). The highest essential oil yield (15.75) were obtained in of 2.5 mg/kg (ZnSO4) and control.Conclusion: Recommended to increase the essential oil yield used 100% humidity and 2.5 mg/kg (ZnSO4) in (Menta piperita L.).

Aim and Background: Legionnaires’ disease is caused by inhalation of contaminated aerosols with Legionella from a variety of water sources. The aim of this study was to characterize the rate of contamination of environmental and hospital water sources by legionella spp in Ahvaz by using standard culture method and polymerase chain reaction (PCR).Materials and methods: A total of 100 water samples were collected from environmental and hospital sources in Ahvaz. After decontamination with Hcl-Kcl treatment, samples were centrifuged and inoculated on to buffered charcoal yeast extract agar (BCYE) and MWY media. In order to confirmation of bacterial colonies as legionella, DNA was extracted and 16S rRNA was amplified using PCR.Results: Out of 100 samples, 15% were positive for Legionella by culture method. Among positive cultures 8% and 7% were obtained from hospital and enviromental water sample, respectively. By PCR all of the positive cultures had 16srRNA and confirmed as Legionella Spp.Conclusion: According to the study results, despite of using purification water 15% of water samples were contaminated with Legionella spp. In order to control and prevention of spread of Legionella spp, disinfection of water sources is necessary.

Aim and Background: Nano-silver as a disinfectant is used for the sterilization of medical laboratory tools, but less attention has been tentative the toxicity of the substance. The aim of this study was to evaluate the effects of Nanosilver on fetal rat liver tissue and determine the expression of MDA levels as an oxidant biomarker and Superoxide dismutase as an antioxidant enzyme. Material and methods: In this study, 16 pregnant Wister rats (four groups of four each) and placebo were treated orally for 18 days with silver nanoparticles of 20 nm in concentration of 125, 750 and 1500 mg / kg / day. The fetuses had been harvested via caesarean after 18 days of gestation to measure the gene expression of SOD in their liver by quantitative RT-PCR method and MDA activity by Satoh protocol in their liver.Results: The highest expression of MDA was detected in the treatment of 750 mg / kg / day, but in comparison to the normal group which showed approximately 68% decrease in concentration. The highest expression of SOD was related to 750 mg/kg/day group which was upregulated 22.4% in compare to normal group, and the lowest expression was shown in 125 mg/kg/day group which was approximately down regulated 5.75 % in compare to normal group.Conclusion: Nano silver particles involved in the process of oxidative/antioxidant system that is caused by arise of free radical production. Antioxidant enzymes such as superoxide dismutase as free radical scavenger was up regulated to detoxify microenvironment. The result of this study revealed that intracellular processes of oxidant/antioxidant system in rat embryo liver were affected by various concentrations of Nano silver particles.

Aim and Background: Nowadays, applying nanocarriers has increased dramatically in drug delivery systems. Niosome is one of the nanocarriers with non-ionic surfactant which increases the stability of drug in the body. Since using cisplatin in treatment of cancer has some side effects, cisplatin was nanoniosomed. Material and methods: To prepare niosome, span 60 and cholesterol were used. Mean diameter of particles was determined by Zeta sizer. Encapsulation efficiency and drug release were measured by spectrophotometry method. Cytotoxicity effect of the formulation on C6 cell line was studied by MTT assay.Results: Mean diameter of particles was verified in scale of nano. Encapsulation efficiency and released in 48 hours were be 43.6% and 97%, respectively. The cytotoxicity effect of nanoniosomal cisplatin was 38.6 mg/ml.Conclusion: The present study showed that to reduce side effects of cisplatin and increase its toxicity, we can use nanoniosomal formulation.

Aim and Background: Amyloids are fibrilar protein structures produced from aggragation of proteins and peptide together. In recent years amyloid fibrils are being noticed as new experimental nanomaterials and from this view the induction of proteins to amyloids could be beneficial. In this study, the fibrillation of bovine serum albumin (BSA) as a model protein is optimized. Materials and Methods: Different pH of 1, 2, 3, 4.7 and 7.4 of Bovine Serum Albumin was prepared in Mixed Citrate–Phosphate buffer and agitated for 0, 24, 48 and 72 hour in 100 RPM in various temperature (40–80oC) for preparing protein fibrils. Then the amount of amyloid fibrils was detected by spectrophotometric congored binding assay.Results: In this research, effect of four variables include temperature, pH, time of incubation and protein concentration was investigated on fibrillogenesis and results were confirmed by congored spectrophotometric method as lmax and absorbance in lmax (Almax) accompanying with transmission electron microscopy. The optimum condition for fibrilogenesis was specified in 5 mg.ml-1 of protein and buffer pH of 3 after 72 hour incubation in 70oC.Conclusion: Simple congored spectrophotometric method could be used as primary test for evaluating protein nanobiofibrils and absorbance in lmax is introduced as a valid indicator in this way.

Aim & Background: Strategy genomics such as analysis of EST sequence, provide to identify markers associated with gene expressed and miRNA related regulatory and metabolic networks. In this study identifying elements associated with genome expressed were examined in Salvia fruticosa leaf trichomes. Material and methods: After collecting and removal of vector sequences, chloroplasts, repetitive, assembly them, were created Singletons and contigs sequences. Using by the BlastX, current hits and frequency and distribution of EST-SSR markers were determined using SSRTools. Related miRNA target sequence using modeling and search were identified in Arabidopsis thaliana.Results: 153 contigs and 777 Singletons was found. BLASTX has revealed 477 unigene with hits. Gene enrichment analysis sequences in different functional groups of the contract. The molecular markers EST-SSRs identified di-nucleotide AT / GA, tri- nucleotides GCC, Tetra- nucleotides TAAT repeated at least four most frequent and penta and hexa-nucleotide repeat of AAGAG and AGTATT, CGTGGT three tandem repeats a times reported. miRNA gene expression were identified related to a number of 40 different genes.Conclusion: EST-SSRs identified in the plant can be used to determine the variation between different species of this genus in the plant family lamiaceae. Two related miRNA (monoterpene synthase) to regulate the expression of genes important in producing terpenoids important to be used in the plant.

Aim and Background: Microbial infections are important challenges to health, and health care officials have major difficulties dealing with them especially regarding their antibiotic resistance. The main aim of this study was to evaluate anti-bacterial effects of Cinnamomun Verum and Ferula Gummosa essential oil on some pathogen bacteria. Materials and Methods: Cinnamomun Verum and Ferula Gummosa essential oil were extracted from its plant. For evaluation of antibacterial effects of the essential oil, disc diffusion method through the measurement of the inhibitory zone. Minimum Inhibitory concentration (MIC) was detected.Results: According to the results of disc diffusion test in Agar, Cinnamomun Verum and Ferula Gummosa essential oil, with a 47 mm and 33 mm inhibition zone, had the greatest antibacterial activity against Staphylococcus Saprophytic us. It had the greatest positive co-action with gentamicin (10 mg) on Escherichia Coli.Conclusion: The results of this study indicate that Cinnamomun Verum and Ferula Gummosa essential oil alone or in combination with antimicrobial agents may be useful in treatment of bacterial infections.

Aim and Background: Due to the extreme growth conditions of most of sulphate reducing bacteria (obligative anaerobic, high temperature, etc.), studying and investigation of them is difficult exclusive Ardebil province climate and its unique hot springs have rarely been studied. The aim of this study was investigation of the potential presence of anaerobic, thermopilic sulphate reducing bacteria in Sarisu hot springs of Ardebil. Material and Methods: In this study sampling was done from Sarisu hot spring. Enrichment, isolation and purification of sulphate reducing bacteria were done in specific media Postgate B and Hungate. Initial identification of bacteria was done by common biochemical tests. Further molecular identification was done on the basis of 16S rDNA riobotyping with the use of universal primers.Results: The studied hot spring contained sulphate reducing bacteria which were able to grow in specific media. Dominant bacteria in these media were gram positive, motile, and had endospore. None of these isolated bacteria had desufoviridin pigment.Conclusion: The result of this study showed the existence of anaerobic thermopilic sulphate reducing bacteria of Desulfotomaculum genus in the studied hot spring.

Aim and Background: Brucellosis is a major cause of zoonosis and in many countries, including Iran, is a public health concern. Developed a new diagnostic method with high sensitivity and minimize the risk of infection in laboratory personnel, is of paramount importance. Including the ways in which these benefits are polymerase chain reaction (PCR). In this study diagnostic methods (culture - molecular) Brucella isolated in samples from patients suspected of having brucellosis in various parts of Tehran are compared and evaluated. Materials and Methods: 100 patients suspected of having symptoms of brucellosis to the laboratory Masoud Tehran, the Bagheat Allah Hospital, Medical Sciences Iran went for blood sampling. Each blood sample medium (Castaneda) inoculation and temperature conditions 370c and %5 carbon dioxide was 7 to 30 days. Brucella strains standard of laboratory reference preparation and were stored under standard conditions. All articles published in the Brucella specific primers collect and studied were specific primers B4/B5 223 bp to reproduction a piece of the gene.Results: Two Brucella strains were isolated from 100 blood samples of the patients with brucellosis by blood culture and DNA isolated by Kit Cinnagen and after the PCR, fifteen were positive. The sensitivity of PCR, %100 specificity and respectively %86, 73.Conclusion: consideration great importance of molecular diagnosis of infectious diseases even at 10 nanograms of bacterial DNA in laboratory samples, also the high sensitivity of the method used in the diagnosis of such diseases on the basis of the results of this study and previous research PCR technique compared with culture method appears to be that the sensitivity is higher. So this could be the reason for the replacement of this method as a diagnostic method in the laboratory.

Aim and Background: Generally, reducing the particle size in the nanoscale particles increases the surface to volume ratio and the impact of the behavior of the material surface atoms compared to the behavior of the mass of atoms. The most important effect of the surface increase is the extreme reactivity of nanomaterials due to high concentration of the surface atoms or molecules in comparison with the mass of the surface atoms or molecules which causes the nanoparticles agglomeration or cluster. To prevent the reactions, a stabilizer should be used to protect particles against abrasion and corrosion.Material and methods: This process is performed using Tetra-Ethyl-Ortho-Silicate. The obtained nanoparticles were characterized by Scanning electron microscopy (TEM), Fourier transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD) and Vibrating Sample Magnetometer (VSM).Results: According to the analysis carried out, particles 40 nm were obtained and silica-coated magnetite nanoparticles were well synthesized. In addition, the nanoparticles magnetic saturation decrease after coating significantly and reduce ability to absorb nanoparticles in magnetic separation process well as.Conclusion: In this article, coating iron oxide magnetic nanoparticles with very thin layer of the silica coating is investigated. According to this method, after stabilization of iron oxide magnetic nanoparticles with silica, in addition to stabilizing the magnetic nanoparticles, magnetic properties of nanoparticles are not decreased.