Modares Journal of Biotechnology

Abstract Introduction: Hypertrophic cardiomyopathy (HCM) is a rare cardiac disorder which can result in sudden death in young people. Molecular studies have been showed that mutations in the beta-myosin heavy chain (MYH7) gene are one of the most common genetic causes of HCM. The aim of this research was to study exons 8, 9, and 30 of MYH7 gene for possible mutation in HCM patients from Chahar Mahal va Bakhtyari Province.
Material and Methods: In this descriptive-laboratory study, DNA was extracted from 27 blood samples by phenol-chloroform method. DNA samples were then used for PCR-SSCP analysis for amplification and identification of mutation. The Suspected cases with possibility of having mutation were sequenced and the results were observed by Chromas software.
Results: 7 suspected cases recognized by PCR_SSCP were sequenced with forward and reverse primers to verify the presence of mutation.

Conclusion: Mutations in these exons do not have a role in establishment of disease in the studied population. However, study greater number of HCM cases and other exons of this gene are recommended to find the relationship between gene and HCM and to gain necessary information for treatment and management of disease.

Abstract Crude oil is comprised of four groups: Saturates Aromatics, Resins and Asphaltenes. Oil pollution has irreversible effects on marine ecosystems. Therefore paying attention to oil pollution and the management of the ports is very important. Biodegradation of oil derivatives is more effective, more powerful and more economically efficient method for remediation in polluted sites rather than physicochemical methods. In this study, for isolation of crude oil degrading bacteria, seawater and mussels were collected from Persian Gulf. Enumeration of bacteria were done in collected samples. Isolated bacteria were identified by biochemical and molecular tests. Crude oil biodegradation for each strain was assessed by spectrophotometer and Gas Chromatography (GC). The results of this study show that the quantity and biodiversity of heterotrophic and crude oil degrading bacteria in Crassostrea gigas mussels was higher than surrounding environment (seawater). Eleven crude oil degrading bacteria were isolated from Persian Gulf, 7 strains were identified biochemically and 2 strains were selected on the basis of higher degradation. These isolated strains were identified as Shewanella and Alcanivorax. The half percentage of oil was removed by these strains in 15 days of incubation. These bacteria could be used for cleanup oil-polluted marine areas after more research and field observation.

Abstract Biodiesel as a clean fuel is renewable, biocompatible and free of polycyclic aromatic hydrocarbons that could be deriving from animals, plants, fungal, algae and bacteria resources. Among these resources, oleaginous fungi due to the high capability to synthesize and accumulation of triacylglycerol are the best source for biodiesel producer. So, in order to providing approaches for increase biodiesel production based on biotechnology, molecular investigation in these organisms could be promising approach, which have been attentioned in this study. In this regard, a precise survey on the related molecular mechanisms led to reveal Malic enzymes as the effective and critical proteins in lipids production and accumulation in oleaginous fungi. Structural characterization of the genes, led to reveal that they are different in the length and GC content as well as they are continuous in the sequence context. Moreover, structural characterization of the enzymes led to determine their localization in the cells, present the functional domains with capability of post-translational modifications in all of them, which are including MAO1_MF, Malic_M and malic. Homologous sequences survey of the enzymes led to introducing fungal species with possible capability for lipid production.structural modeling of the selective malic and malic like enzymes led to provided suitable models in structure and quality in function with binding affinity to malate. In general, the results of this study, while introducing suitable fungal species for securance of biomass, led to reveal effective enzymes with special features that could be useful in tracing the capable strains or transgenesis modification.

Abstract Cellulase is one of the industrial enzymes which its production and utilization is increasingly taking into consideration due to global heed to second-generation bioethanol production. Cellulase produced by different organisms such as fungi, bacteria, insects, and plants. With increase in utilization of this enzyme and need for reduction in the enzymes price for production of second-generation bioethanol, the production of recombinant enzyme has been considered noticeably.
In this study, by investigation of corn steep liquor as nitrogen source and second carbon source after glycerol, a new medium is designed based on SYN6 salt medium then biomass and endoglucanase II production by methylotrophic yeast was optimized. Experiments designed by one-factor and response surface methodology used for optimization.
Results showed that optimum conditions for biomass and endoglucanase production are 5.5% (w/v) and 6.15% (w/v) of corn steep liquor respectively. New optimized conditions increased 41.4% and 69.7% for biomass and recombinant enzyme production respectively.

Abstract Production of recombinant proteins in Escherichia coli has been very common in recent decades. Many studies and experiments have been done in order to optimize the production and expression of recombinant proteins in E.coli. One strategy is using high cell density to increase recombinant protein production such as β-NGF in the cell. Therefore, in this study for the first time bacterial cell culture in high cell density was done using glycerol and yeast extract as carbon and nitrogen sources and MgCl2 as a growth effective factor. Also the effects of overnight culture conditions on bacterial growth were evaluated. Meanwhile culture conditions were optimized using response surface methodology (RSM) and the optimum conditions were as follows: 18/23 g/lit glycerol, 14.44 g/lit yeast extract and 10mM MgCl2. Also the obtained results indicated that the 14 hours incubation at 37 °C and 180 rpm were optimum conditions for the overnight culture. Our results showed that the rate of cell growth and recombinant β-NGF production in optimized condition is significantly higher than in basic medium.

Abstract The aim of this study was to evaluate the performance of Horizontal Sub-Surface Flow Constructed Wetland (HSSFCW) for the removal of Cr by Phragmitis australis and to assess the effect of the plant, bed material, hydraulic loading rate and hydraulic retention time on system performance. In this study, 12 cells of pilot system were built in parallel way and in dimensions of 50 × 75 × 200cm at the end of water treatment plant in Birjand University. Temperature and pH were same in all the cells. The results showed that the removal of Cr in cells containing plants was higher than cells without Phragmitis australis. Change of the bed material from coarse texture to fine-grained texture will make significant increase in the average percentage of Cr removal(the removal percent insamedischarge 100(l/d), in fine and coarse-textured plant cells was%31.87 and %24.61respectively and inlack ofplant'scells,with fine and coarse textures was%31.25 and %14.92, respectively). By increasing the retention time of 1 to 5 days, Cr concentration and consequently the removal rate were increased. These results are indicative of the positive effect of HSSFCW system in the presence of Phragmitis australis in removal of heavy metals such as Cr, therefore to remove heavy metals from wastewater, cultivation ofPhragmitis australis and fine-grained texture is recommended.

Abstract One of the molecular mechanisms of alcohol-induced toxicity is mediated by oxidative stress. We investigated the protective effects of orally administered vitamin C (VC) in different doses on oxidative damage in rat eyes induced by chronic ethanol intake.
Eight groups of rats were treated for 30 days: control (C), VC (50, 100 and 200 mg/kg), ethanol (4 g/kg) and, ethanol + VC (50, 100 and 200 mg/kg).Eyes were then removed for analysis of oxidant/antioxidant markers including superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and malondialdeyde (MDA).
Ethanol decreased the activities of SOD, CAT and GPx in the eye. These changes were accompanied by enhanced lipid peroxidation measured as increased MDA levels. Although VC (50 and 100 mg/kg) restored antioxidant enzymes activity and lipid peroxidation, there were still significant differences compared to control animals. However, 200 mg/kg VC clearly prevented the pro-oxidant and antioxidant imbalance. Interestingly, the highest dose of VC produced a potent inhibition of lipid peroxidation and improvement in antioxidant defense enzymes compared to other doses in non-alcoholic rats.
Oral administration of VC 200 mg/kg for 30 days prevented redox imbalance induced by chronic ethanol exposure in rat eyes by enhancing the activities of antioxidant enzymes and inhibiting lipid peroxidation. Relative to other chemical medications, vitamin treatments may be free of major side effects; therefore this antioxidant vitamin may provide a potential alternative for prevention of ethanol toxicity which deserves consideration and further examination.

Abstract Background: wrongs in experiments and laboratories are inevitable particularly if used non-standard methods and tools. In routine disc diffusion test used optical method for pouring media in petri dishes that create non negligible errors in experiments. Goal of this research is design and produce a standard petri dish for this tests and eliminate the wrongs.
Methods: According CLSI standards about sensitivity tests medium standard thickness is 4 millimeters. According these data designed a standard petri dish that determined this standard thickness exactly.
Results: Results of this research showed existing Petri dishes produce wrong and different reports about non-growth haloes even for identical antibiotics that cause to antibiotic unfit administration significantly (P<0.05). In addition, medium waste rate was 33-50% for different petri dishes.
Conclusion: Designed standard petri dish standardized disc diffusion tests and other sensitivity tests and makes accuracy, non- growth haloes uniformity in identical tests, and antibiotics fit choose. Also decrease medium waste rate significantly.

Abstract Nanotechnology is a principally attractive area of research related with production of nanoparticles of variable sizes, shapes, chemical compositions and their possible application for human being benefits. Creation, manipulation and utilization of metallic nanoparticles, because of reduction of materials dimensions, affect the physical properties and results in displaying extraordinary thermal, optical and electronic properties of nonmaterial. The biological approaches to synthesis of nanoparticles are better than chemical and physical procedures because of low energy and time expenditure.
In this study the possibility of production of nano-silver particles from dried flower buds of Clove was investigated and antibacterial and anti-fungal activities of produced nanoparticles were studied by diffusion disc and well methods. The displayed UV-visible spectra, with a wavelength of 300 to 600 nm, identifies formation of silver nanoparticles, whenever the colorless initial acclimated mixture turned to brown. The centrifuged powder samples were examined using scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) methods. Based on the results of this study, produced silver nanoparticles were spherical in the range of 27 to 69 nm and showed effective antimicrobial activity against Bacillus subtilis, Escherichia coli, Staphylococcus aureus and Saccharomyces cerevisiae. Therefore clove can be used as a biological source for the synthesis of nanoparticles in an industrial scale with a very low cost.

Abstract Aim and Background: Amylloid fibrils are filamentus protein aggregates derived from various proteins and peptides. They can be distinguished from other type of features according to their appear shape and electron microscope images, also by dye binding methods, which can indicate induced cross beta structures. Amyloid fibrils are correlated to creating general disease, amyloidosis. Disease such Alzheimer, Parkinson, diabetes type II, and others disease which in each of them, the special kind of protein subjected to form amyloid or amyloid like fibrils. A variety of proteins which they are not converted to amyloid fibrils invivo, can be transform to amyloids in special unstabilizing conditions.
Materials and Methods: Congored spectrophotometric method, ThT fluorescence and CD Data was used for fibril formation assay and Transmission Electron microscopy was used for final affirmation of fibrils.
Results: results shows that maximum amyloid formation was in 5 mg.ml-1 protein concentration, 50 ºC and 7.4 buffer pH.

Conclusion:
With the new approach obtained from the kappa casein, amyloid fibers can be introduced as new nanomaterials, Thus the results, given the diverse applications of nanomaterials, can affirm process optimization of amyloid production from accessible and inexpensive protein in milk.

Abstract Newcastle disease is a fatal viral disease which is highly contagious that affects most species of birds and is a major economic threat in the poultry industry. Both the HN and F glycoproteins of Newcastle disease virus (NDV) are essential for pathogenicity and virus infectivity. This study describes immunization of DNA vaccines encoding the HN, F or both the genes of New castle disease virus. In our previous study, the antigen expression of the insert genes has been validated in vitro by Western Blotting and Indirect Immunosenest. In this study, ELISA and HI analysis of the in vivo experiment on SPF (specific Pathogen Free) chickens showed the induction of humoral responses by the DNA vaccines. Our finding indicated that twice vaccination with pDNA was able to elicit significant antibody titers (P< 0.05) by either monocistronic (pIRES/HN and pIRES/F) or bicistronic (pIRES/F/HN) plasmid, after one week of second pDNA vaccination (booster). The results proposed that DNA immunization of chickens at second vaccination had enhanced the antibody response successfully. Also, it revealed that vaccination with the co-expression plasmid pIRES/HN/F can induce a stronger antibody response than vaccination with pIRES/HN or pIRES/F alone.

Abstract in recent years, interest in use of plant sources due to function of Phytochemicals and antioxidant compound in maintenance of human health is increased, phenolic and flavonoid compounds are one of important compounds of plants that have antioxidant effects. Aim of the present study is to examine and comparison of phenolic and flavonoid compounds and antioxidant capacity of different organs of saffron, that every year a huge amount of them are wested during the processing of stigma. In this study, different parts of saffron were extracted by methanol (80%), then the amount of total phenol and flavonoid compounds was assayed by means of Folin-Ciocalteu and and Aluminium choloride methods respectively. The antioxidant activity was measured by DPPH free radical reduction. According to the results of this study, The highest content of total phenolic(6.43 mg GAE g-1 DW) and flavonoid (1.33 mg RU g-1 DW) was observed in stigma compared to other organs. The result of DPPH test also showed higher antioxidant capacity of stigma in comparison to other organs. Comparison of phenolic compounds in various organs showed that the content of these compounds and antioxidant activity could be different related to type of organs. Also, the higher antioxidant capacity in stigma and tepal compared to leaf and corm could be as a result of more phenolic compounds in these organs.