NEW CELLULAR & MOLECULAR BIOTECHNOLOGY JOURNAL
Year:2013 | Volume:3 | Issue:12|Papers Count:16

Aim and Background: Using sensitive and specific laboratory techniques such as PCR is necessary on diagnosis of hepatitis which caused by hepatitis B virus (HBV). Common serological methods didn’t have the capability of quick and accurate detection of infection detection thought molecular methods such as PCR are effective tools for evaluation prevalence of HBV. A different result for the reason of not being standard is one of the disadvantages of this rich molecular technique. One of the most significant preventives in widespread usage of diagnostic PCR is not having proper PCR internal controls. The aim of this study is designing and constructing plasmid internal control (IAC) for identification of PCR inhibitors and afterward usage in diagnostic laboratories.Materials and methods: In this study for constructing the internal control, PCR specific primers for HBsAg gene of hepatitis B virus were optimized then sensitivity and specificity were determined. Also Composite Primers for HBV-IAC were designed and amplified by PCR then cloned. Minimum number of IC in each PCR reaction was examined through dilution and PCR results with IC.Results: By using special primers HBV PCR product was 262bp and IAC-HBV product was 660bp which had desirable different in sizes. Minimum number of IC in each reaction was 1000. Maximum of PCR sensitivity with IC for hepatitis B virus DNA was determined as 16 million Particles. In specificity test with different factors no non specific product was observed.Conclusion: For Molecular detection of HBV using an internal control can recognize the errors and will standardize this sensitive technique.

Aim and Background: Paclitaxel is one of the drugs used in breast cancer. Use it as a side effect to weakening the bone marrow and peripheral neuropathy causes. Recently been shown that nanotechnology can be used to further reduce side effects, increase the efficiency of treatment. One of the nano-carriers for drug delivery, are niosomes. In this study nano paclitaxel niosomal with the ether injection methods and reverse-phase evaporation were prepared and studied. Aim of this study use of nanoparticles niosomal containing paclitaxel and cytotoxic effects on breast cancer cell lines.Materials and methods: In this study, in order to improve the therapeutic effects and reduce side effects paclitaxel, drug with both methods by ether injection and reversed phase evaporation, was preparate nano-niosomal pegilation. Span 20, cholesterol, polyethylene glycol l2000 Dalton, paclitaxel, solution of MTT (0.5 mg/ml) was purchased from Sigma. Ethanol and isopropanol and diethyl-ether from Merck Co. and culture RPMI 16-40 purchased from Invitrogen Co. and MCF-7cells were obtained from the Pasteur Institute Cell Bank of IranResults: After a description of resulting nanoparticles indicated the manufacturing method, combined with polyethylene glycol exert a positive effect on the properties of nanoparticles. We Could encapsulation amount of paclitaxel in niosomal nanoparticles to be dramatically raise.Conclusion: In this study were identified by using nanotechnology and niosomal nanoparticle can be appropriate formulation of the paclitaxel “drug” prepared, Be more effective than the free drug the dosage for treatment is reduced and this reduces the side effects are On the other side, is a low-cost treatment.

Aim and Background: Anaplastic carcinoma is the most dangerous form of thyroid cancer. It is a rare and aggressive form of thyroid cancer. It grows very rapidly and does not respond to therapy. In this study the effect of Silibinin on the metabolic activity of a cancerous cell line (8305c) of anaplastic thyroid carcinoma was investigated. Silibinin is a flavonoid that is the major active constituent of the extract of the Milk thistle seeds. Silibinin has been demonstrated to have strong pleiotropic anti-cancer effects against various human carcinoma cells.Materials and methods: The 8305c cell cultures were treated with Silibinin at 25, 50, 75, and 100 μmol in the MTT plates. After 24 and 48 hours, the metabolic activity of the treated cells was studied using the MTT assay. The MTT assay is a colorimetric method for determination of cell viability and cytotoxicity effects.Results: The results of this study show that Silibinin has significant cytotoxicity effects on the 8305c cell line. The treatment of the cells with Silibinin at the given concentrations resulted in significant decrease of metabolic activity in a time and dose dependent manner.Conclusion: From the results obtained by this study It may be concluded that Silibinin significantly reduces the growth of anaplastic thyroid cancerous cells. More investigation is underway to reveal the mechanism by which Silibinin prevents cancer metastasis.

Aim and Background: The objective of this research was to evaluate the effects of Bio fertilizer (Azosprillium spp bacteria) and Salicylic acid on growth and development characteristics of Ocimum basilicum L. plant under water stress.Materials and Methods: The experimental design was factorial arranged in a completely randomized block design with three replications. Treatment were combination of 3 levels of water stress (100% FC as a control, 60% FC and 30% FC) and Bio fertilizer (inoculated and non-inoculated) and Salicylic acid (0 as control and 0.75 mM).Results: Data variance analysis showed that morphological characteristics was significantly (p<0.05) affected by drought levels, Salicylic acid, Bio fertilizer and their interaction. The effects of various treatments on shoot and flower number, dry and fresh weight of root and shoot, Relative water content (RWC) and electron leakage was significant at 1%.Conclusion: Application of salicylic acid and Azosprillium inoculation increased growth and development of Basil under water stress.

Aim and Background: Exopolysaccharides (EPS) produced by lactic acid bacteria (LAB) play an important role in taste and texture improvement of fermented dairy products. Additionally, they may be important to the health promoting properties of LAB through host immune stimulation. The aim of this study was exopolysaccharide production of 16 Lactobacillus strains isolated from traditional dairy products of Iran. It was performed to isolate and identify both bound and released exopolysaccharides.Materials and methods: Twenty eight gram positive, catalase negative lactobacillus strains were selected based on their association with fermented dairy products of diverse origin. Colonies were assessed for both microscopic and macroscopic features. The isolates were further evaluated for release and bound EPS; we used a phenol/sulfuric acid method. Results were compiled using a glucose standard curve.Results: Level of bound EPS production ranged from 4-364mg/l and release EPS from 87.25 to 1519 mg/l. The EPS producing strains isolated from dairy products may have properties of emulsification, stabilization, and film formation, to improve flavor, appearance and texture in foods. The EPS produced by LAB may have beneficial effects on human health such as cholesterol-lowering ability, immune modulating, and antitumor activities and prebiotic effects.

Aim and Background: Teucrium polium L. belongs to the genus of Teucrium and Labiatae family. It has been used in folk medicine to treat some diseases because of its useful effects. This research is done in order to compare chemical composition of essence of Teucrium polium in different habitats for the first time.Materials and methods: The flowering aerial parts of Teucrium polium were collected in 9 habitats in Mazandaran province and dried. The habitats were named G1-G9. The essential oils of the dried flowering aerial parts obtained and were analyzed by the combination of GC and GC-MS.Results: The major compounds upper 10% contained: G1: Sabinene (28.4%), GermacreneD (11.2%), G2: GermacreneD (19.64%), Sabinene (18.75%), G3:Sabinene (28.58%), Cis: Sesquisabinenehydrate (12.64%) G4: Sabinene (15.47%), GermacreneD (12.8%), G5: GermacreneD (18.07), Sabinene (12%) G6: Sabinene (29, 3%), a-Pinene (10.1%) G7: E-Nerolidol (23.21%) Sabinene (20.14%), G8: Sabinene (31.35%), E-Nerolidol (15.81%) Valerianol (10.23%) G9: Sabinene (26.93%), b-Pinene (21.23%).Conclusion: G6, G9, G3 and G4 were known as the best habitats because of quality of essence.

Aim and Background: Micro-organisms that can accumulate lipid more than 20% of their biomass are called oleaginous. Microbial lipid has high similarity with plant’s oil; the only difference between them is that microbial lipids are rich in polyunsaturated fatty acids. Synthesized lipid by the yeast has potential of being used in pharmaceutical industry for technical purposes or as nutritional supplements. The importance of microbial triacylglycerols is the high amount of polyunsaturated fatty acids with 16 and 18 atoms of carbon which is important from nutritional point of view.Materials and methods: In this study the amount of lipid production by standard and native yeast strain on glucose, xylose and glycerol as carbon sources was evaluated. After selecting the type of carbon source, the effect of MgSO4 on lipid production was investigated by one factorial method. Following these steps, optimization of other parameters such as carbon and nitrogen concentration, temperature, agitation rate, pH and time of incubation was done with Taguchi method. FTIR Spectroscopy was used for analysis and verification of produced oil, in this study.Results: The yeasts Yarrowia lipolytica DSM70562 and Geotrichum BL have reached to lipid production of 7.35 g/L and 5.35 g/L, respectively. The most effective factor in optimization process of evaluated yeasts was the concentration of carbon source. This factor for the evaluated standard and native yeasts strains was 41.37% and 63.57%, respectively. The results of lipid analysis by FTIR Spectroscopy technique showed remarkable similarity between extracted oil from yeast strains and related standard (triolein).Conclusion: According to the results obtained by Taguchi design, lipid production in evaluated standard and native yeasts increased 2.94 g/L and 1.22 g/L, respectively after optimization. This shows the high efficiency of using Taguchi design in optimization of medium condition. Therefore, by using Taguchi design we can do the optimization process, which is the necessary and effective step of all industrial process for increasing productivity and reducing the costs, as best as possible.

Aim and Background: Wide application of nanoparticles in different areas of medical science has been growing faster than before. Small size makes them highly active. Therefore this gives them chemical, physical and biological unique properties. Nanoparticles have no problem crossing through biological barriers in body. Hence, these particles may be utilized in order to target drugs and some other materials. Wide application needs to investigate their cytotoxicity effects on target cells and adjacent cells. In this study we aimed to analysis effect of nanoparticles on invitro.Materials and methods: Related articles in this review were obtained from PubMed, Google and Elsevier Science using the key words nanoparticles, cytotoxicity and differentiation.Results: Based on the results of this study, nanoparticles are able to leave both positive and negative effects. In vitro experiments indicate that the main mechanism of action is based on changing levels of intracellular ROS.Conclusion: The results of reported studies have shown that nanoparticles are able to alter intracellular ROS levels and affect growth and differentiation cells.

Aim and Background: Micro-organisms that can accumulate lipid more than 20% of their biomass are called oleaginous. Microbial lipid has high similarity with plant’s oil; the only difference between them is that microbial lipids are rich in polyunsaturated fatty acids. Synthesized lipid by the yeast has potential of being used in pharmaceutical industry for technical purposes or as nutritional supplements. The importance of microbial triacylglycerols is the high amount of polyunsaturated fatty acids with 16 and 18 atoms of carbon which is important from nutritional point of view.Materials and methods: In this study the amount of lipid production by standard and native yeast strain on glucose, xylose and glycerol as carbon sources was evaluated. After selecting the type of carbon source, the effect of MgSO4 on lipid production was investigated by one factorial method. Following these steps, optimization of other parameters such as carbon and nitrogen concentration, temperature, agitation rate, pH and time of incubation was done with Taguchi method. FTIR Spectroscopy was used for analysis and verification of produced oil, in this study.Results: The yeasts Yarrowia lipolytica DSM70562 and Geotrichum BL have reached to lipid production of 7.35 g/L and 5.35 g/L, respectively. The most effective factor in optimization process of evaluated yeasts was the concentration of carbon source. This factor for the evaluated standard and native yeasts strains was 41.37% and 63.57%, respectively. The results of lipid analysis by FTIR Spectroscopy technique showed remarkable similarity between extracted oil from yeast strains and related standard (triolein)Conclusion: According to the results obtained by Taguchi design, lipid production in evaluated standard and native yeasts increased 2.94 g/L and 1.22 g/L, respectively after optimization. This shows the high efficiency of using Taguchi design in optimization of medium condition. Therefore, by using Taguchi design we can do the optimization process, which is the necessary and effective step of all industrial process for increasing productivity and reducing the costs, as best as possible.

Aim and Background: Epidemiological and experimental studies of animals indicated that flavonoid rich diets can affect positively on decrease of the prevalence of neuronal degenerations e.g. Alzheimer and Parkinsonism diseases. In the basis of the positive function of crataegus extract on decrease of oxidative factors and other helpful characteristics of this extract, the present research examined crataegus extract effect on passive avoidance learning of male wistar rat.Materials and methods: 24 rats with 250-300 gram weight were divided into three main groups: 1- control (intact) 2- sham (solvent receivers) 3- Crataegus extract. The extract was administered daily (100mg/kg) for two weeks by gavages’ method. Passive avoidance learning test was conducted by shuttle box system. One day after acquisition, retrieval test was done. For this, the animals were placed in a light compartment and after 30 seconds guillotine door were opened and the time that was consumed for entering to dark component was recorded. Data were analyzed by SPSS software with one way ANOVA and Tuckey test.Results: The results indicated that there is no difference between intact and sham groups at the time of entering to dark component but there was significant increase between extract and intact groups (P<0.001) and decrease between them for entering dark component (P<0.001).Conclusion: Results of this study showed that treatment by Crataegus extract that is flavonoid rich, resulted in the recovery of learning and memory.

Aim and Background: Galanin is a potent factor regulating appetite and energy balance. Galanin injection to the nucleus involved in food intake stimulates food intake. On the other hand, negative energy balance is accompanied with epinephrine and cortisol level. In this study cortisol and epinephrine effect on galanin secretion in rats fed with different levels of their energy requirement was investigated.Materials and methods: 45 male wistar rats (300-350 g, 15 per group) were fed a diet containing 100%, 50% and 25% of their energy requirement for 10 days. Afterwards each rat after 48 h prolonged fasting was anesthetized and then cannulated in the carotid artery for injection and blood sampling. Animals were divided into 3 treatment groups that received (3 mg/Kg BW) either EN, Cortisol or combination of those two (0.1 mg in 1 ml of PBS). Galanin and glucose levels were analyzed before (in time of 0) and 30, 60 and 120 min after injection.Results: In 50% and 100% food restricted group fasting, galanin levels fell after EN injection. (p≤0.05). Cortisol just reduced galanin in 50% food restricted group (p≤0.05). In contrast, the group that had 25% food restriction didn’t show any response to cortisol, EN and combination of EN and cortisol injection (p>0.05).Conclusion: These results indicate that injection of EN suppress starvation-induced secretion of galanin in normal (100%) and starved (50%) rats. And the galanin secretion response to EN might be affected by weight loss as it is not suppressed in starved (25%) rats.

Aim and Background: In recent years the threat of acquisition of antibiotic resistance caused by overuse of antibiotics is growing. The purpose of this study was to survey the pattern of antibiotic resistance and evaluate the multi-drug resistant Escherichia coli and analyze the plasmid profile of these clinical strains.Materials and methods: This study was performed on 83 clinical Escherichia coli, obtained from four hospitals (i.e. Imam Khomeini, Alborz, Kasra, Qaem) and two private laboratories (Razi and Kavosh), of Karaj City. Isolates were identified as Escherichia coli based on standard biochemical tests. The isolates were screened for Antimicrobial susceptibility test by 20 antibiotics using Kirby-Bauer disk diffusion method. Plasmid profiles of these isolates have been analyzed by alkaline lysis method.Results: Antibiotic resistance patterns were observed as follows: Nitrofurantoin 8.4%, Tetracycline 63.8%, Amoxicillin 83%, Ofloxacin 30%, Levofloxacin 30%, Co-trimoxazole 59%, Amikacin 18%, Imipenem 25.3%, Choloramphenicol 16.9%, Cephalexin 61.4%, Gentamycin 28.2%, Norfloxacin 33.7%, Nalidixic acid 56.6%, Cephalothin 48.2%, Ciprofloxacin 32.5%, Ceftazidime 27.7%, Ceftriaxone 37.3%, Ceftizoxime 16.9%, Cefotaxime 38.5%, Amoxycillin/Clavulanic acid 68.7%. Of 83 isolates 92.77% (77) isolates were resistant to more than two unrelated drugs. Of 83 isolates, 65 isolates, (78.3%) showed the presence of plasmid.Conclusion: The results of this study indicate a growing resistance pattern to the first line antibiotics, prescribed for urinary tract infection. High presence of plasmids in these strains, play an important role in multidrug resistance in these bacteria.

Aim and Background: Paraffins are high molecular weight hydrocarbons and in the reservoir, under high-temperature/pressure conditions remain in a state of equilibrium. However, as the crude oil is pumped out to the surface, temperature and pressure are reduced. Below the cloud point, the paraffinic hydrocarbons precipitate out of the crude oil and form deposits on the oil production systems. These problems are usually mitigated by thermal fluid treatments, pigging or using chemicals and solvents. While most of these remedies are in use, they have certain disadvantages. Moreover, these remedial options are expensive. A microbial intervention to prevent paraffin deposition is a non-hazardous and economically viable approach. This investigation was aimed at finding a bacterial strain being able to effectively degrade tetracosane.Materials and methods: Soil samples were collected from oil contaminated soil near Tehran refinery. Samples were transferred into 40 ml sterile mineral salts medium containing tetracosane as the sole source of carbon and energy. By a multi step enrichment and screening technique a bacterium was isolated and by morphological characteristics, biochemical tests and phylogenetic methods, it was identified as the Mycobacterium genus.Results: This strain was able to use tetracosane as the sole source of carbon and energy. Our results showed the selected strain grew well in pH range of 4-9 and temperature range of 30-40oC with optimal growth at 35oC, had an acceptable growth at 5% NaCl and was able to degrade 80 percent of the tetracosane within 24 days.Conclusion: According to the rate of tetracosane decomposition by this bacteria under conditions listed, its application can be profitable in preventing paraffin deposition. Further investigation in physical model is required to confirm the ability of the strain in wax deposit mitigation from oil wells.

Aim and Background: Gastro-intestinal cancers are the most common kind of cancers all over the world. They are multifactorial disease and both genetical and environmental factors are involved in their progression. Adenocarcinoma is the most common type of stomach cancers. Brevinin-2R is a novel antimicrobial and anticancer peptide that is isolated from the skin secretions of a family of frogs called Rana. The 25-amino acid long peptide shows anticancer activity and induces antimicrobial activity. Brevinin-2R is a 25 amino acid long cationic peptide that shows very low hemolytic activity against red blood cells. In this paper we study cytotoxicity of brevinin-2R onto AGS cell line.Materials and methods: AGS cell line was incubated with different concentrations of peptide during different periods of time, cytotoxicity of peptide and viability of AGS cell line was assessed by MTT-assay. The experiment was repeated three times and the average viability values were calculated.Results: Our findings show that viability of treated cells decreased consistently with increasing concentrations within the periods of time.Conclusion: Because of low hemolytic activity, Brevinin-2R is a good specimen in drug designing. Brevinin-2R triggers cell death by means of activation of autophagy and shows anticancer activity against AGS malignant cell lines.