Modares Journal of Biotechnology

Abstract Multiple sclerosis (MS) is commonest demyelinating disease among young adults. It demonstrates neurological dysfunctions in females more than males. Gonadal hormones have essential roles in maturation and differentiation of neurons and glial cells in both genders. The role of some gonadal female hormones such as progesterone has been well characterized on demyelination and remyelination in animal models. The role of androgens on neural system development and myelin maturation were identified. We previously observed that castration decreases the brain resistance against demyelinating insults and also reduces the subsequent repair. As the optic nerves and chiasm demyelination shows the hallmark characteristic in MS, inducing demyelination in optic apparatus, we have tried to find whether the effects of elimination of male gonadal hormones using gonadectomy could show the same, more or less changes in patterns of demyelination and repair comparing demyelinated females? Thus, to evaluate these alternations castrated male and female rats were compared by using visual evoked potentials and histological assessments on 2,7,14, and 28 days post lysolecithine (LPC) injection. Interestingly, we observed demyelination was started 2 day post lesion (dpl), reached to a maximum level at 7 and 14 dpl and then it partially but significantly reversed on 28 dpl. Demyelination and subsequent repair processes in both gonadectomized and female groups were shown almost the same patterns temporarily and in quality. Elimination of gonadal androgens could cause the male animals to undergone the same shape of de/remyelination compared female ones. In conclusion, differences between male and female demyelination and remyelination are substantially depending on male gonadal androgens. This work could be useful for understanding of the effects of sex hormones on demyelinating diseases and could offer fundamental information for repairing therapies in Multiple Sclerosis.

Abstract Abstract Introduction: Measles virus (MV) belongs to the morbilivirus genus of paramyxoviridae family, and has single stranded, negative polarity, non segmented RNA genome. Method: In this research, the total RNA was extracted of measles virus (AIK-C) vaccine strain. The extracted RNA was immediately used in reverse transcription reaction to generate cDNA. The 1st strand cDNA was used to amplify the F gene by specific primers in a reaction PCR. The PCR product with the expected size of 1662 bp was cloned into expression plasmids pET-22b(+) and pET-28a(+). The recombinant plasmids were transformed into competent E.coli DH5α cells and clonies were screened with direct PCR. The recombinant plasmids were extracted by Alkaline lysis and were compared with non- recombinant plasmids in molecular weight. Results: Recombinant plasmids were digested with Nde I and Hind III restriction enzymes. The DNA band with an approximate size of 1662 bp was detected on 1.5% agarose gel. The recombinant plasmid pET-28a(+) was sequenced, comparison of this sequence with the coding sequence F protein of measles virus (AIK-C) in Genbank (AF266286) was revealed high degree of homology and showed that F gene is highly conserved. Conclusion: It was showed that F gene is highly conserved. Thus F gene is important for studing in order to produce recombinant vaccine.

Abstract Evaluation of Liver Enzymes’ Level in Blood Serum of Crystal Addicts Introduction: Crystal or dimetamphetamine is one of the harmful substances which are imported to Iran in the last few years. Crystal is made of pure heroin. Its harmful effects are more than heroin. Addiction to crystal can produce psychological and organ damage. In order to find out which part of body could be affected, examination of enzymes is useful. Liver is one of the organs which can be infected by drug abuse. Liver function test can be used to show these harmful effects. The aim of the present study is to exanimate of the effects of crystal abuse on liver enzymes such as Aspartat transaminase, Alanine transaminase and Alkaline phosphatase in crystal abusers. Materials & Methods: The current study is a case-control and cross-sectional study on 105 crystal addicted as case group who had referred to addiction treatment center for the first time and 100 healthy people as control group. The activity of enzymes was measured by callorimetery-spectrophotometry method. Results were analyzed by T-test exam and SPSS-16 software. Results: results show that using crystal increases the level of AST, ALT and ALP and there is a significant relationship between crystal abuse and the level of ALP (p=0.027). In addition, there is a significant relationship between duration of crystal use and the level of ALP (p<0.05). Conclusion: comparing with AST and ALT levels, ALP level is more affected by crystal abuse.

Abstract We present a method to predict the flexible and rigid regions based on sequence. We use the free energy of two consequent amino acids to define a factor for distinguishing flexible regions from the rigid ones. Using statistical analysis of this free energy, we assign a normalized number between zero to one hundred which we call it flexibility number. Taking the effects of up to four neighbors of an amino acid, into account, resulted in an efficient prediction of flexible and rigid regions of a protein.

Abstract In order to the investigation on the effects of methyl jasmonate on the secondary metabolites of Calendula officinalis L., the plants were grown in a modified Hogland (1/2) solution and were treated with 50 and 100 µm methyl jasmonate. The results indicated that jasmonate treatment decreased lignin content of root but increased lignin content of shoots, in comparison with those of the control plants. However, total content of wall-bound phenolics of roots increased and those of shoots decreased by jasmonate treatment. Anthocyanin content of shoot, decreased by methyl jasmonate. Likewise, flavonoid contents of shoots in both treatments were lower than those of the control plants. Lipid peroxidation rate of roots and shoots did not show significant changes in jasmonate treatments, compared to the control plants. In both control and jasmonate treated plants, α- Cadinol was the most abundant essential oils component. Induction of α-Muureloene with specific antifungal properties in the jasmonate treated plants and increase of it along with increase of jasmonate concentration, suggested that jasmonate can be used in order to induce modifications in secondary metabolism pathway of Calendula officinalis, resulting to produce desired medicinal compounds.

Abstract Oxidative Stress is fast becoming the nutritional and medical buzzword for the 21st century, and imPlicated in a growing list of diseases, from cataract to cancer. Oxidative stress occurs when the amount of free radicals in the body exceeds its pool of available antioxidants. Some researches indicate that margarine contain Trans fatty acids as major sources of dietrary free radicals and oxidative Stress. But other researches has been shown that because of supplementation of mairganie with some antioxidant such as alpha-carotene, beta-carotene, vitamin C and specially vitamin E, significantly decreased free radicals and oxidative stress and increased the levels of the supplied antioxidants in plasma. In this study, we examined the effect of The use fortified margarine on oxidative stress markers in young male, rats. To evaluate the margarine effect, 20 wistar rats were divided into 2 groups and selected as control and marganine groups. In second groups margarine were added to their chow as 15% (w/w). After 4 months blood samples were obtained, Then plasma levels of antioxidant, lipid peroxidation and thiol group of its, has been measured. This experiment showed that margarine intake has a positive effect on markers of oxidative stress, and consumption of antioxidant supplied in a full-fat margarine and consumed as a part of normal diet, effectively increases the blood levels of antioxidant and eliminats of free radicals.

Abstract Lentiviruses are considered one of the most effective recombinant viruses for gene transfer to mammalian cells and tissues. In this study, the potential of HIV-1-based lentiviral vector to deliver transgenes into avian cells was examined. We co-transfected human embryonic kidney cell line HEK-293T with three lentivirus vectors called transfer, packaging and envelope vectors. We collected the supernatant from transfected cells 24 and 48 hours post-transfection and filtered them immediately. Then we subjected the filtered supernatant to Amicon protein columns for concentration purposes. Centrifugation removed a larger part of the supernatant presumably free of viruses and left behind a small volume of darken solution full of virions. We thereby produced a 500-µl-volume of virus stock. Various dilutions of this stock were added to chicken liver cell line LMH. The initial sign of infection appeared within 48 hours and by 96 hours post-infection 100% the LMH cells positively expressed transgenes. Our results indicated that the human HIV-1-based lentivirus vectors are capable of transducing and transferring foreign genes into chicken cells. Given the need for a high-titer virus stock for successful target cell transduction, our results indicate that the filtration method of virus concentration is able to produce high virus titer and is cost-effective and less time consuming than ultracentrifugation or other traditional methods.