فصلنامه تازه های بیوتکنولوژی سلولی مولکولی
پیاپی 23 (تابستان 1395)

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 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.
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 ∆∆Ct method. The average ∆Ct of CK19 and GAPDH was calculated in each cell line as well as the ∆Ct of GAPDH as a calibrator in different cell lines was quantitatively reported.
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.
Expression of CK19 is the same, in the luminal A breast cancer cell lines. Molecular markers could be used for cell Lines classification which is significant in breast cancer research and it is helpful in detection and treatment of the disease.

Aim and Various factors, such as sexual diseases, spinal cord injuries and radiotherapy after cancers, cause insufficient or lack of sperm production and lead to men infertility. Today, treatment of many diseases has been made possible by therapeutic cell technology with mesenchymal stem cells. Meanwhile, mesenchymal stem cells of human umbilical cord Wharton's Jelly, have been considered as a source that is abundantly available and not rejected by immune system. The first step in using these cells is to confirm they are stem cells. Therefore, in this study stemness of these cells were investigated by expression of CD34, CD44, CD105 genes.
Stem cells were isolated from umbilical cord Wharton's jelly and cultured. Total RNA was extracted from cells of the third passage. After synthesis of cDNA, expression of some genes was evaluated by RT-PCR technique. The mesenchymal stem cells were exposed to different doses of BMP4, conditioned medium obtained from testicular cells of newborn mice and a layer of PLLA Nanofiber. Then differentiation of mesenchymal stem cells to germ-like cells was evaluated by different techniques.
The results showed that these cells have considerable potential for proliferation and expression of CD105, CD44 genes before differentiation, but the expression of CD34 gene, a marker for hematopoietic cells, was not seen. The exposure of these cells to differentiation medium after the third passage, leads them toward germ-like cells.
Laboratory culture and proliferation of germ-like cells derived from other sources of stem cells such as umbilical cord Wharton's Jelly is a new method for initial research about development of germ cells. Umbilical cord stem cells with expression of stemness specific genes can be used for treatment of men infertility as a new strategy.

Aim and 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. Therefore, the use of techniques applied to measure the enzyme activity is important. Several methods by using various Ops substrates can be designed.
Material and 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.
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.
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 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 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.
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 significant (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) was 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) was also obtained in 2.5 mg/kg (ZnSO4) and control.
Recommended to increase the essential oil yield used 100% humidity and 2.5 mg/kg (ZnSO4) in (Menta piperita L.).

Aim and 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).
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 confirm bacterial colonies as legionella, DNA was extracted and 16S rRNA was amplified using PCR.
Out of 100 samples, 15% were positive for Legionella by culture method. Among positive cultures 8% and 7% were obtained from hospital and environmental water sample, respectively. By PCR all the positive cultures had 16s rRNA and confirmed as Legionella Spp.
According to the study results, despite of using purification water 15% of water samples were contaminated with Legionella spp. In order to control and prevent spread of Legionella spp, disinfection of water sources is necessary.

Aim and 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 In this study, 16 pregnant Wister rats (in four groups of four) 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.
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 comparison to normal group, and the lowest expression was shown in 125 mg/kg/day group which was approximately downregulated 5.75 % compared to normal group.
Nano silver particles involved in the process of oxidative/antioxidant system caused by arise of free radical production. Antioxidant enzymes such as superoxide dismutase as free radical scavenger was upregulated 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 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 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.
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 µg/ml.
The present study showed that to reduce side effects of cisplatin and increase its toxicity, nanoniosomal formulation can be used.

Aim and Amyloids are fibrilar protein structures produced from aggregation 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.
Different pHs of 1, 2, 3, 4.7 and 7.4 of Bovine Serum Albumin were prepared in Mixed Citrate–Phosphate buffer and agitated for 0, 24, 48 and 72 hour in 100 RPM in various temperatures (40–80 ºC) for preparing protein fibrils. Then the amount of amyloid fibrils was detected by spectrophotometric congored binding assay.
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 λmax and absorbance in λmax (Aλmax) accompanying with transmission electron microscopy. The optimum condition for fibrillogenesis was specified in 5 mg.ml-1 of protein and buffer pH of 3 after 72 hour incubation in 70ºC.
Simple congored spectrophotometric method could be used as primary test for evaluating protein nanobiofibrils and absorbance in λmax is introduced as a valid indicator in this way.

Aim and Genomics methods, such as EST sequences analyses, provide the ability to identify markers associated with expressed genes and miRNAs of regulatory and metabolic networks. In this article, the identification of elements related to the genome expressed in leaf nodes of Salvia fruticosa was examined.
A number of 1465 EST sequences related to the genome library expressed in the leaf node cells of plants of genus Salvia, from Lamiaceae family, were received from the NCBI database. After collecting and removal of sequences related to vector, chloroplasts, repetitive sequence and aligning them, Counting and Singleton sequences were created. Using BlastX search, the existing hits as well as the frequency and distribution of EST-SSR markers were determined by using SSR Tools. The miRNA related to the target sequences were identified using the modeling and searching in Arabidopsis plant.
A number of 153 Cotig sequences and 777 Singleton sequences were found. The BLASTX revealed 477 unigenes with hit. The gene enrichment analysis put the sequences in various functional groups. Among molecular markers identified in the EST-SSRs, the AT/GA di-nucleotides, GCC tri-nucleotides and TAAT tetra-nucleotides with at least four repeats have the highest frequency. In case of penta and hexa-nucleotide repeats, the AAGAG, AGTATT and CGTGGT were reported as three subsequent repeats. For the expressed genome, 40 miRNAs related to different genes were identified.
The 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 important related miRNAs to regulate the gene expression of monoterpene synthase enzyme in the production path of important terpenoids in the plant can be used.

Aim and 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.
Cinnamomun Verum and Ferula Gummosa essential oil were extracted from its plan. For the evaluation of antibacterial effects of the essential oil, disc diffusion method through the measurement of the inhibitory zone. Minimum Inhibitory concentration (MIC) was detected.
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, manifested the greatest antibacterial activity against Staphylococcus Saprophyticus. It had the greatest positive co-action with gentamicin (10 μg) on Escherichia Coli.
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 the treatment of bacterial infections.

Aim and Due to the extreme growth conditions of most of sulphate reducing bacteria (obligate 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, thermophilic sulphate reducing bacteria in Sarisu hot springs of Ardebil.
Material and 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.
The studied hot spring contained sulphate reducing bacteria 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.
The result of this study showed the existence of anaerobic thermophilic sulphate reducing bacteria of Desulfotomaculum genus in the studied hot spring.

Aim and Brucellosis as zoonosis diseases is a public health concern in many countries, including Iran. New detection methods with high sensitivity are of importance and minimize the risk of infection in laboratory staffs. One of these methods is polymerase chain reaction (PCR). At the present study, culture and molecular methods for detection of Brucella obtained from patients with suspected brucellosis in various parts of Tehran were compared and evaluated.
100 blood samples were obtained from patients with suspected brucellosis who referred to the Massoud Laboratory, Baghiatallah and Hospitals affiliated to Iran University of Medical Sciences. Each sample was inoculated in Castaneda medium and incubated in a chamber with 5% CO2 at 370C for 7 to 30 days. DNA extraction from samples and PCR carried out using commercial kit and standard procedure, respectively.
Two samples were positive from culture of 100 blood samples, while after the PCR, fifteen samples were positive. The sensitivity and specificity of PCR determined to be %100 and 86.73%, respectively.
Culture of Brucella spp. is not always successful due to their growth difficulty; also the growth and culture require viable organisms. Many bacteria are lost during sample taking or using antibiotics. This may cause the sensitivity of culture to be low in comparison with PCR.
conclusoin:PCR compared with culture method appears to be more sensitive and so this method might be the reason for the replacement of this method as a detection method in the laboratory.

Aim and Generally, reduction of 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.
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).
According to the analysis carried out, particles measured 40 nm were obtained and silica-coated magnetite nanoparticles were well synthesized. In addition, the nanoparticles magnetic saturation decrease significantly after coating and reduce ability to absorb nanoparticles in magnetic separation process well as.
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.