NEW CELLULAR & MOLECULAR BIOTECHNOLOGY JOURNAL
Year:2017 | Volume:7 | Issue:27 |Papers Count:17

Aim and Background: Lac promoter, a regulatory gene in Escherichia coli, is a prokaryotic expression system for recombinant Interferon production. This promoter is induced by lactose or IPTG, its chemical analog. In this study, the effect of lactose and IPTG on the expression of gene was compared.Materials and Methods: Escherichia coli BL21 (DE3) harboring Interferon b -1b gene were cultured and induced by the addition of IPTG and / or lactose. The yield of produced Interferon protein was measured.Results: The yield of Interferon b-1b protein increased by using 0.2% lactose and 0.5 mM IPTG simultaneously as inducers in comparison with each inducer alone and it was equal to 34% of the total cell proteins.Conclusion: In this research, with simultaneous presence of Lactose and IPTG, in comparison with each one of this inducers alon, a higher yield of the interferon beta 1b protein at a lower cost was produced.

Aim and Background: In order to evaluation of seed priming on Moderating of salinity effect on germination and primary growth of cucumber in pot planting, an experiment carried out by Factorial design with RCD at three replication in greenhouse of Zahedan Jihad Keshavarzi in 2012.Material and methods: The main factor was salinity at three level consider EC=2, EC=4 and EC=6 mmohs/cm and second factor was seed priming in four level consider control, KNO3, KCl and NaCl.Results: The variance analysis of data showed that salinity had meaning effect on the all of investigated traits. The most germination percentage with 97.58% obtained in control treatment. The maximum biomass as well as in this treatment with 55.26 gram. The seed priming had meaning effect on the all traits except leave number, stem diameter and the time need for emergence. The most germination with 88.11% obtained in KNO3 treatment this treatment as well as with 43.92 gram had the most biomass. The correlation effect of salinity × priming had no significant effect.Conclusion: According to this results cucumber cultivation not recommended for Areas where the irrigation water is saline.

Aim and Background: Cannabinoids are known to attenuate memory in both humans and animals that effects of this compounds act through their receptors. These compounds can affect on the memory and learning through the hippocampus. Acetylcholine, as a neurotransmitter, plays fundamental role during process of learning and memory and donepezil can be potentiation effect in cholinergic synapse On this material. In this research the effects of donepezil, URB597 “potentiation endocannabinoid system " and effect of both them on the memory in mice was investigated.Materials and Methods: In this experimental research 40 male mice weighting 20-35 g were divided to the 4 groups including control, URB597 (0.1 mg/kg, i.p), donepezil (0.5 mg/kg, i.p.) and URB597 & donepezil. Mice to the first day received in the Shuttle box electrical trial then at the second day intraperitoneal administrate of drugs 15 minutes before the retrieval test. Two analysis parameters, delay the first time in the dark chamber (STLr) and sum of to remain time in the dark chamber (TDC).Result: Analysis of data indicated, that there was not significant difference in received number of trials. But there was significant difference between groups control and URB597 (p<0.001) in STLr. Also there was significant difference URB597 (p<0.001) in comparison by control and compound URB597 & donepezil groups in TDC and also there was significant difference in compound URB597 & donepezil (p<0.005) group in comparison to control group.Conclusion: Result of this experiment shown that URB597 led to memory impairment and administration of donepezil, as cholinergic system potentiation, has improvement affect on the destructitive memory, so indicate interference of both system led to memory reinforcement.

Aim and Background - Streptococcus pyogenes is the cause of many important human skin-diseases such as Psoriasis. Infections can be associated with the release of bacterial virulence factors the same as, A, B, C exotoxins. Study on existence of speA, speB and speC genes in Streptococcus pyogenes isolated from Psoriasis patients was this research’s aim.Materials and Methods- From 60 Psoriasis patients, by biochemical methods was isolated 48 Streptococcus pyogenes. Then DNA was extracted from bacteria and used PCR for identification of speA, speB and speC genes.Results: Frequency of these genes in Streptococcus pyogenes were for, sepA (58.3%), sepC (33.3%) and sepB (100%). Frequency of strains that carried sepA and sepB genes, were 58.3%, sepB and sepC genes, were 33.3% and sepA, sepB and sepC genes, were 20.8%.Conclusion: Study results was identified that speA, speB and speC related to Psoriasis. This study also signifies bacterial factor between different effective causes for Psoriasis such as genetic and physiologic factors are main, and in comparison to studies carried out elsewhere in the world Streptococcus pyogenes is an important factor for causing of Psoriasis.

Aim and Background: Zinc is one of the metals that preserve the structure and function of many tissues in the body. Its effect is concentration-dependent. It can disrupt the vital process of cellular respiration. This study is aimed to investigate the interaction of this metal in the function and structure of peroxidase as one of the most important oxidoreductase enzymes in cellular respiration process, experimentally and theoretically.Material and methods: In order to extract kinetic parameters using UV-visible spectroscopy techniques and molecular docking modeling, horseradish peroxidase activities were plotted against time and various concentrations of zinc. Absorbance spectrum of the enzyme was obtained in the presence and absence of zinc, to determine the structural change, which are responsible for enzyme inhibition.Results: The results showed that Zn2+ strongly inhibited peroxidase activity in a reversible uncompetitive-noncompetitive pattern with IC50 370 mM. Moreover, Zn2+ enhanced the lag time and decreased steady state rate of the enzyme. UV-visible spectrum and molecular docking analysis indicated that Zn2+ binding site is distinct from the active site and transformational change and reduction of electron transfer ability of porphyrin ring were occurred in the presence of Zn2+. Conclusion: We propose that excess amount of zinc induces inhibition in peroxidase activity and possibly causes disorder in cellular respiration and electron transport. It is also believed that it could be useful in drug design for cancer cells.

Aim and Background: RNA isolation is difficult in plant that contain large amounts of polysaccharides and polyphenol compounds that bind and co-precipitate with RNA. Therefore, in this study eleven different methods were evaluated to extract RNA from leaves of apple rich in phenolic compounds.Material and methods: The quantity and quality of extracted RNA were determined by 1.2% nanodrop spectrophotometric at absorbance of 230, 260 and 280nm, agarose gel electrophoresis and cDNA synthesis and RT-PCR analysis by specific primer for apple Actin were performed. Results: The results indicated that methods of Djami-Tchatchou et al., (2012) and Rajakani et al., (2013) were the most suitable methods for RNA extraction from apple leaves. The yield of total RNA extracted by these methods varies from 753.6 to 800.2 and 362.1 to 452.6 ng/ml in apple leaves , respectively. Distinct bands of 28S and 18S ribosomal RNA were observed in electrophoresis.Discuss: In two protocols, ratios of A260/A280 and A260/A230 ranged from 1.99 to 2.03 and from 2.07 to 2.17, respectively This is indicative of a high RNA purity and the absence of contamination with polyphenolic, protein and polysaccharide compounds.Conclusion: The high quality for RT-PCR reaction by two methods showed the extracted RNA can be used with assurance in downstream molecular techniques.

Aim and Background: Milk thistle (Silybum marianum) is a medicinal Thorny Shrub of the Family of Compositae, Which Contain Silymarin. It is Useful for Liver Disease and Concer Prevention and Treatment. And it is Also Beneficinal for Lowering Blood Cholestrol. The Medicinal Importance of This Species Necessitates Studying in This Field.Materials and Methods: In this study, three species from Moghan, Fridonkenar and Noorabad Mamasani of the Family of Compositae were evaluated in terms of Karyotype Silybum marianum. Karyotypic studying population of this species, Alfabromonaftalyn Was Used as a pre-treatment, chromium oxide and formaldehyde as a fixator, NaOH for hydrolyz and hematoxylin for coloring.Results: After microscopic studies, the statistical analysis and preparation of tables and the karyotype, it Was determined that All of the populations are diploid and have a basic chromosome number x = 17 and the Chromosomes Place in Metacentric and Submetacentric Distances. This Means That The Centromere Place in Chromasomes is Opproximately in Middle.Conclusion: According to the Shape of The Chromosome and the Centromere Place and TF% Parameter. It Can be Concluded that the Silybum marianum is Aless Developed Plant With a Symmetric Karyotype. Also From the Developmental Point of View the Noorabad Mamasanis Population Was More Developed Than the Other Two Populations. And the Studied Populations Belong to One Species and the Transformation and Separation of Species Were not Observed.

Aim and Background: Listeria monocytogenes is known as a food-borne pathogen responsible for causing disease in humans. This bacterium that cause meningitis, abortion and food poisoning in humans. Industrial sauce as a matter of day-to-day offer food consumption among the general population have found. This food, suitable for food poisoning. Therefore contamination with Listeria monocytogenes is a threat to health, especially those with suppressed immune systems. The aim of this study is molecular detection of listeria monocytogenes in industrial sauce in Iran.Materials and Methods: 25 Industrial sauces were collected from Iran markets. DNA samples extracted by standard phenol-chloroform method. PCR test optimized and detection of limits (LOD) and specificity evaluated. All of the samples tested with optimized PCR test. Results: PCR test Optimized and 226 bp amplicon was observed in 2% agarose gel electrophoresis. LOD test was obtained 10 genome/reaction and in specificity test, amplicon was not observed with none of microorganism DNA was tested. After PCR testing on 25 sauce samples, 4 were positive for Listeria monocytogenes.Conclusion: PCR technique using specific primers very carefully and faster than traditional methods is for the identification of Listeria monocytogenes. The results show that the PCR technique is a good way to monitor microorganisms in food products such as sauces.

Aim and Background: Osteosarcoma is the most common cancer among adolescents. Curcumin has induction sensitivity effect to cancer cells against chemotherapy and anti-tumor. In present study, the sustained- release niosomal carrier containing Curcumin were prepared to improve anticancer effect of Curcumin against osteosarcoma.Material and methods: Anionic prepared niosomal vesicle are contained 70 % (molar ratio) of Tween 60, 30% (molar ratio) of Cholesterol in the presence DSPE-mPEG 200. Prepared nanoparticle was characterized and the cytotoxicity of free Curcumin was evaluated in comparing of niosomal Curcumin on MG-63 cell line using MTT assay. A suitable kinetic model was also suggested in order to predicting drug release. Results: Results showed the size diameter, encapsulation efficiency, zeta potential and poly disparity index are 242 nm, 95.2%, -38 mv and 0.17, respectively. Prepared niosomal particles are controlled-release and the cumulative release is 43.87 % during 96 hours. Peppa’s model indicates good confirmation with experimental results. Cytotoxicity results showed 0.9-fols improvement in killing cells efficiency for niosomal Curcumin in comparing free Curcumin.Conclusion: The niosomal carrier containing Curcumin is sustained-release, mono-disperse with high encapsulation efficiency and suitable size diameter. The results of surface charge evaluation confirmed that prepared nano-carrier is anionic and it can effectively be used in the treatment of bone cancer.

Aim and Background: Heavy metals are such as pollutant nature, under functions such as mining and industrial waste to entering the environment. Heavy metals including mercury, cadmium, nickel, arsenic, copper, lead, cobalt, silver and chrome. Some heavy metals are needed for cell growth in small amounts and it is toxic in high concentrations. High concentrations of heavy metals are created serious problems for the life of plants, animals, fishes and finally to humans who are the main consumers of these food sources. Identifying bacteria resistant to heavy metals plays an important role in relation to environmental pollution and eventually its cleanup. Therefore, in this study, the molecular heavy metal resistance in Escherichia coli strains isolated from surface water has been studied.Materials and Methods: In this study, 120 samples of surface water in Tehran in the spring and summer of streams, rivers and sewage were collected and using capped sterile Falcon transported to the laboratory immediately. The first samples were enriched in the BHI broth culture. Then to see colonies of microorganisms, were cultured on BHI agar linear and were incubated at 37oC for 24 hours. To isolate strains from other bacteria grown in culture medium, used Eosin methylene blue (EMB) specific medium. Using the disk plate and serial dilution in the tube method, the resistance of bacteria to different concentrations of salt heavy metals zinc and cadmium sulfate was studied.Results: Only 40 (33.33%) E. coli strains was isolated. The minimum inhibitory concentration (MIC) was reported for zinc sulfate salt was 12.5 mM and for cadmium sulfate salt was 10 mM.Conclusion: By comparing heavy metal zinc and cadmium resistant in E. coli strains was found that the amount of zinc in surface water study is more than cadmium. So the zinc resistant in Escherichia coli strains is more. Generally increase in heavy metals in surface water results in increased resistance of Escherichia coli strains to heavy metals.

Aim and Background: Multiple sclerosis (MS) is the most common autoimmune disease of the central nervous system among young adults. Many factors such as environmental factors and genetic background are involved in the development of MS. miRNAs in nervous and glial cells have an important role in post-transcriptional levels. Any change in their performance by polymorphisms and in their expression in nervous system can be grounds for degenerative diseases such as MS. The purpose of this study was to investigate the role of polymorphisms miR-23a C> T(rs 3745453) in Iranian population susceptibility MS.Materials and Methods: In this study, 134 patients with MS were selected, and 142 healthy individuals were chosen as controls. Tetra ARMS-PCR was used to for finding genotype. Results: No difference was observed in dominant (P>0.05; OR:0.77; 95%CI: 0.48-1.23) or recessive (P>0.05; OR: 0.80; 95%CI: 0.36-1.75) genotype distribution miR-23a (rs3745453) in patients with MS, as compared to controls.Conclusion: This study was performed for the first time in Iran, unlike previous studies that indicated association between this polymorphism and occurrence of MS in patients, in this study was not correlation between polymorphisms miR23a (rs3745453) and MS disease in a sample of Iranian population.

Aim and background: Asthenozoospermia is one of the most common reasons of infertility in men, which is known as low or no movement of sperm. The purpose of the present study is to clarify the relationship between polymorphism rs1893316 of gene Catsper1 with Asthenozospermia in men.Materials and methods: The present molecular study, After filling in the consent form by both 50 control and 50 experimental groups, 2ml blood was taken, then DNA isolated through salting out method and sequencing procedure was used to specify polymorphism. Result: In studying polymorphism rs1893316 C>T CC genotype frequnency and TT genotype frequnency was considerably higher in control group than experimental one(P<0.05). In hetrozygotic genotype CT frequency, there wasn't a significant difference between experimental and control group(P>0.05).Discussion: Catsper1 channel physiologically is limited to sperm movement path, following it, the release of calcium ion and imbalance in ionic density leads to sperm movement increase regarding both it's strength and speed.Conclusion: There is a meaningful relation between polymorphism rs1893316 gene Catsper1 with Asthenozoospermia. The result may change with a change in genetic treasury in the participants or meaningful change in population size.