فصلنامه تازه های بیوتکنولوژی سلولی مولکولی
پیاپی 12 (پاییز 1392)

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. 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. 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. For Molecular detection of HBV using an internal control can recognize the errors and will standardize this sensitive technique.

Aim and 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. 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 Iran. 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. 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 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. 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. 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. 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 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. 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). 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%. Application of salicylic acid and Azosprillium inoculation increased growth and development of Basil under water stress.

Aim and 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 because of comparison of chemical composition of essence of Teucrium polium L. in different habitats at the first time. The flowering aerial parts of Teucrium polium L. was 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 combination of GC and GC-MS. The major compounds upper 10% were 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%)، α-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%)، β-Pinene (21. 23%). G6، G9، G3 and G4 were known as the best habitats because of quality of essence.

Aim and 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 using 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. 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 selection the type of carbon source، the effect of MgSO4 on lipid production was investigated by one factorial method. After 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. 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). 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. So 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 Wide application of nanoparticles in different area of medical science has been growing faster than ago. Small size makes them high active. Therefore this gives them chemical، physical and biological unique properties. Nanoparticles will have not much problem to cross through biological barriers in body. Hence we can use these particles for targeting drugs and some materials. Wide application needs to investigate their cytotoxicity effects on target cells and adjacent cells. In this study we tried to analysis effect of nanoparticles on invitro. Related articles in this review were obtained from Pub Med، Google، Elsiver science using the key words nanoparticles، cytotoxicity and differentiation. Base on the results of this study nanoparticles are able to leave both positive and negative effects. invitro experiments indicate that the main mechanism of action is based on changing levels of intracellular ROS. The results of reported studies have shown that nanoparticles are able to alter intracellular ROS levels and affect growth and differentiation cells.

Aim and In this study، Preparation and isolation of Iranian Mesobuthus eupeus venom toxic fractions has been performed. Its venom has different toxin same the other scorpions which affect on ion channels. For studying of these toxins، the venom was isolated during 3 stages gel chromatography، DEAE anion and CM cation exchange chromatography respectively. At each stage، the fractions were identified with UV spectroscopy and collected with fraction collector then the fractions injected to 2 normal mice and the toxic fraction with killing effect selected for next stage. The results of injected fractions showed that the fractions F3، F31، F316 and F319 were toxic. To determine the location of venom concentration in different organs can help to design proper antivenom.

Aim and Epidemiological and animal experimental studies indicated that flavonoid rich diets could affected positively on decrease of the prevalence of neuronal degenerations e. g Alzheimer and Parkinsonism diseases. In basis of the positive function of crataegus extract on decrease of oxidative factors and other helpful chararacteristics of this extract، the present research examined crataegus extract effect on passive avoidance learning of male wistar rat. 24 rats with 250-300 gram weight have been divided to three main groups: 1-control (intact) 2- sham (solvent receivers) 3-Crataegus extract. This extract has been administered daily (100mg/kg) for two weeks by gavages'' method. Passive avoidance learning test were achieved by shuttle box system. One day after acquisition retrieval test has been done. For this، animal was putted to light compartment and after 30 seconds guillotine door was 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. were indicated no difference between intact and sham groups for 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 to dark component (P<0. 001) Results of this study showed that treatment by Crataegus extract that is flavonoid rich، has been resulted to recovery of learning and memory.

Aim and Galanin is a potent factor regulating appetite and energy balance. galanin injection to the nucleus involved in food intake، stimulate food intake. On the other hand، negative energy balance is accompanied with epinephrine and cortisol level. In this study was investigated cortisol and epinephrine effect on galanin secretion in rats fed different levels of their energy requirement. 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 recived (3 µg/Kg BW) either EN، Cortisol or combination of those tow (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. 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 has 25% food restriction didn’t show any response to cortisol، EN and combination of EN and cortisol injection (p>0. 05). These results indicate that injection of EN suppress starvation-induced secretion of ghalanin in normal (100%) and starved (50%) rats. And the galanin secretion response to EN might be affected by weight loss as it doesn''t be suppressed in starved (25%) rats.

Aim and 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. This 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 that being able to effectively degrade tetracosane. 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. This strain was able to use tetracosane as the sole source of carbon and energy. Our results showed the selected strain grow well in pH range of 4-9 and temperature range of 30 - 40 °C with optimal growth at 35°C، had an acceptable growth at 5% NaCl and was able to degrade 80 percent of the tetracosane within 24 days. 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.