فصلنامه تازه های بیوتکنولوژی سلولی مولکولی
پیاپی 11 (تابستان 1392)

Aim and “Asparaginase” is an enzyme that plays a key role in curing acute lymphoblastic leukemia (ALL). Due to its antineoplastic property، it is widely used in chemotherapy. The goal of this study was to explore and introduce extracellular L-asparaginase enzymes produced by Bacillus spp. which potentially possess more desired serological properties and less side effects comparing to those of common commercial types. The Bacillus spp. isolated from protein rich soils were cultured in modified M9 media. Asparaginase producing colonies were differed based on color change in media. Then the produced enzyme was purified and its activity was evaluated. Also its molecular weight was measured. Those Bacillus spp. which were capable of producing enzymes with desired activity، were identified using molecular method. New L-asparaginase producer Bacillus strains were identified and the general properties of extracted enzymes were studied. As extracted L-asparaginase from different bacteria has shown different anti-cancer effects، exploring for microorganisms that produce this enzyme، is one of the main ways that would lead us to an enzyme with ideal medical aspects.

Aim and The Wdhn13 gene codes the LEA 2 protein family. LEA proteins were identified and discussed in wheat and cotton as a proteins cumulative first time in the late embryo period. The primary structure of LEA proteins، including early charged domains and non-charged of polar amino acid residues non hydrophilic is that LEA proteins are being implicated on hydrophilic. in this research، the Phylogenetic and bioinformatics relationship of Wdhn13 in 8 wheat species was investigated. The HindIII (AAGCTT)، AluBI (AGCT) and TaqI (TCGA) restriction enzyme were used for molecular analysis and by biological CLC main workbench 6 software the Insilco and out of vitro were applied in sequence Phylogenetic analysis result based on UPGMA algorithm showed that studied wheat species were divided into 3 clusters. The first cluster includes 4 wheat cultivar NCBI database’s sequence، T. aestivum CV Sardari، T. durum CV shosh، T. durum CV borojerd. The T. urartu was located in the second cluster alone. The third cluster includes T. aestivum CV gonbad and T. dicocoides. The results indicated a relationship among some wheat species.

Aims and scope: Several viruses so far have been reported to be the cause of mosaic and yield losses in French bean (Phaseolus vulgaris L.) in Iran. Among these، BCMV، and BCMNV are more important. During a growing season، French bean plant samples were selected with various symptoms of mosaic، vein clearing، leaf rolling، leaf and stem necrosis were collected from bean cultivation fields. The samples were tested serologically against the two viruses using DAS-ELISA and Tissue blot technique with the specific polyclonal antisera obtained from commercial sources. To confirm the Elisa results، selected Elisa positive samples tested applying IC- RT-PCR using virus specific polyclonal antibody and a specific set of primers designed from virus cp-genes region. The cDNA obtained were multiplied in termocycler machine. The results of agarose gel electrophoresis of approx. 373bp and 683bp band were obtained for part of the cp genes of BCMV and BCMNV. Using IC-RT-PCT both viruses were identified from samples of Arak، Neishabour، Zanjan and Karaj provinces. BCMV and BCMNV are widespread in almost all bean growing regions of Iran and often present in mixture. IC-RT-PCR test for diagnosis and identification of these two viruses is proved to be more sensitive than Elisa test. In this technique combination of a specific antisera and a specific set of primers help in distinguishing the two viruses in mixture.

Aim and Chronic Obstructive Pulmonary Disease (COPD) is caused by inflammation and destruction of a part of lung and usually appears after age 50. Considering the muscle weakness، energy limitation and problem with oxygen consumption in COPD patients، we analyzed the mitochondrial cytochrome c oxidase (COX) gene to find the relationship between COXI and COPD disease. We studied 65 patients (71±8 years old) who were clinically stable and 50 healthy people (56±4 years old) as a control group. DNA was extracted and prepared according to the standards. Isolated DNA was used for amplification of COXI gene. In this study، screening four new polymorphisms using ARMS Real time PCR was performed in COXI. PCR conditions were optimized empirically for screening 4 polymorphisms by ARMS Real time PCR. Significant difference polymorphisms between COPD patient and normal control was found The light note of polymorphism was found at 6060 locus، which is the only polymorphism that leads to an amino acid change. Increasing the value of this polymorphism in patients with COPD than normal individuals can make this polymorphism a risk factor in COPD

Atherosclerosis and diseases derived from vascular calcification are one of the important causes of mortality in the patients with rheumatoid arthritis (RA). Although several studies have assessed in this case، however، there are no many results on relationship between those factors and bone biomarkers and their correlation with vascular calcification. The aim of this study was designed to investigate the comparison of serum levels of matrix Gla protein (MGP) and osteoprotegrin (OPG) and their relationship with the severity of disease activity (DAS28) in women with rheumatoid arthritis. 45 female patients with rheumatoid arthritis and 45 matched healthy women as control group were selected. Serum levels of matrix Gla protein (MGP) and osteoprotegrin (OPG) by ELISA، and serum high sensitivity CRP (hs-CRP) by torbidometery method were measured. Results indicated a significant increase in serum OPG (p= 0. 001) and hs-CRP (p= 0. 001) in the patient group were compared to control group. No significant differences were found in the level of serum MGP (p=o. 302) between patient and control groups. Significant positive correlations were found between hs-CRP and erythrocyte sedimentation rates (ESR) (p=0. 001، r=0. 487)، between hs-CRP levels and the degree of disease activity of 28 (DAS28) (p=0. 001، r=0. 570) and between ESR and values of (DAS28) (p=0. 007، r=0. 448) in patient group. The present study confirms that increasing OPG level could possibly decrease inflammatory responses by reduction of cytokines level in women with rheumatoid arthritis; and may reduce the bone destruction. Also، reduction in MGP could result in vascular calcification and vessel wall damage in RA patients.

It has been suggested that histamine has modulatory influence on anxiety-related behaviors both in animals and humans. On the other hand، GABA (A) receptor modulating drugs such as benzodiazepines have been used to treat anxiety disorders. Co-administration of the aforementioned drugs would be of high interest in terms of anxiety effects. Method & Materials: In the present study، we investigated the anxiety-related effects of individual and combined intraperitoneally administration of histamine and diazepam، a benzodiazepine agent. To do so، anxiolytic indices of Open Arm Times (%OAT) and Open Arm Entries (%OAE) were measured using an elevated plus-maze test of anxiety on rats. Intraperitoneal histamine administration 20 (mg/kg) decreased %OAT، but not locomotor activity، showing an anxiogenic response. Subcutaneous administration of diazepame at dose of 1 mg/kg increased %OAT and %OAE، but not locomotor activity، showing an anxiolytic response. Subcutaneous administration of diazepame، at doses of 1 mg/kg before histamine (20 mg/kg) increased %OAT and %OAE but not locomotor activity، thus showing an anxiolytic response. Our results indicated that the anxiogenic effects induced by histamine in the intraperitoneal could be reversed in the presence of diazepam.

Aim and Based on confirmed reports، it is clear that breast cancer is one of the most prevalent diseases among women. Hydroxyureais one of the important drugs used in chemotherapy. despite its therapeutic propertie، this drug has many adverse effects in long-term، this study was performed in order to use the Nano drug delivery technologyusing liposomes drug deliveryfor reducingthe adverse effects and increasing therapeutic index as well. A certain ratio of Phosphatidylinositol choline and cholesterol was synthesized in order to prepare the nanoliposome followed by adding the hydroxyurea drug. The mean diameter of nanoliposomal hydroxyurea was determined by zeta sizer system. The percent of drug released from liposome was performed by dialysis. Also، encapsulation efficiency of nanoliposomal hydroxyurea was calculated. Ultimately the cytotoxicity of formulation was probed by MTT assay. The mean diameter of nanoliposome hydroxyurea and also and its encapsulation efficiency were recorded 402. 5 nanometers and 70. 836 % respectively. The pattern of drug release from nanoliposomes using dialysis method showed that the drug release of nanoliposome drug within 28 hours was equal to 25. 85 %. This study showed that the cytotoxicity effect of nanoliposomal drug is more than that of the released drug.

The most common disease of abnormal carbohydrate metabolism، diabetes mellitus is due to defects in insulin secretion or failure in performance or both will develop. It is known that melatonin may be involved in sugar metabolism. Therefore in this study meet the compounds of indole ring (melatonin precursor) on non-diabetic and diabetic rats were studied. Diabetic and non diabetic rats were divided into 4groups. Control group: diabetic and non diabetic rats that received daily water and normal food. Sham group: diabetic and non diabetic rats that received daily water and normal food with 0. 5 ml grape seed oil. Exp1&2 groups: diabetic and non diabetic rats that received daily 4 mg/kg indole، 4mg/kg5-hydroxy indole 2- carboxylic acid in 0. 5ml oil respectively. Diabetizing with alloxan was performed using 150 mg/kg dose with single IP injection. In this study، blood sugar، average urine and water consumption، blood insulin levels were measured، and pancreas as well as histological studies were performed. After treatment for 30 days in the exp1D showed a blood sugar levels، water consumption and urine significantly at p <0. 001 decreased While insulin levels also increased in this group (p <0. 001). While the exp 2D have less effect on insulin levels، blood sugar and the water consumption and urine. Histology also showed that the indole in the exp1D repair pancreatic tissue. this study it was found indole may have beneficial effects in decrease blood sugar and increase insulin synthesis and repair of the pancreas.

Aim and Peroxidases functions in plants range from cell wall modulations to hormone regulation، defense mechanisms، indoleacetic acid degradation and stress adaptations. During diverse stress conditions، peroxidases protect plant against the intrinsic and environmental disturbances. In this study، the plant corms were prepared in July. Corms were used as the source of the enzyme extraction and this extract was used for further study. The results were shown for the dormant and waking corms; increasing concentrations of guaiacol above the 70 Mm decreased the peroxidase activity. With respect to the kinetics results، it seems that at least two isoforms of the peroxidases exist in saffron corms and so each isoform could take part in some different processes in saffron growth and development.

Backgrounds and Ambient particulate matter (PM) is considered to be an important indicator of outdoor air quality. Many adverse health effects are associated with high concentration of particulate matter in ambient air. Exposure to pollution due to bioaerosols is an almost inescapable feature of urban living throughout the world. Exposure to ambient air microorganisms is associated with wide range of adverse health effects. The aim of the present study was to determine the concentration of particulate matter and airborne bacteria in Ahvaz ambient air and investigation of the environmental parameters effects on airborne bacteria concentration. Air sampling for presence of bacteria was conducted using a microbial air sampler (Quick Take-30، SKC، USA). Tryptic Soy Agar which is a specific media for bacterial culture was used as a media. The number of cultivable bacteria on TSA media was counted using colony counter apparatus and was expressed as colony forming units per cubic meter (CFU m-3). The mean concentration of PM10، PM2. 5، PM1 and airborne bacteria at four sampling sites were 149. 35، 46. 58، 25. 6 µg/m3 and 446. 67 CFU /m3، respectively. The highest concentration was observed in the Day month (21 December to 21 June). The lowest concentration was detected in Bahman (21 June to 21 February) month. It was found that the more the human activities، the denser the urban environment، the lower vegetations، the higher the bacterial concentration، which bacterial concentrations have reverse relation with temperature and UV index.

Aim and Brucella spp. is a gram negative coco bacillus which has different hosts including human، cow، goat، sheep and horse. Brucella spp. is transferred via milk and dairy products causing a Malta fever in human. Serological methods for Brucella spp. detection do not have sufficient sensitivity and accuracy; in addition، they have great false positive and negative results. The aim of this study is to assess the prevalence of Brucella spp. in dairy products with Nested PCR technique and evaluation of specification and sensitivity of this technique. In this study 208 samples from dairy products including: 57 raw cow milk، 34 pasteurized cow milk، 28 pasteurized cheese، 23 traditional cheese، 33 raw goat milk and 33 raw sheep milk samples، were collected from Tehran and suburbs. DNA was extracted using DNP kit (Cinnagene، Iran) from samples. After optimization of first and second round of PCR for Nested، with the help of each specific primer، sensitivity and specificity technique was evaluated. The studies showed، Among 57 samples of raw cow milk، 19 cases were PCR positive. In 34 pasteurized milk samples 10 cases were PCR positive، from 28 pasteurized cheese samples only 8 cases، from 23 traditional cheese DNA samples 14 cases، out of 33 samples of raw goat milk 21 cases and finally from 33 samples of raw sheep milk 19 cases were PCR positive. With respect to this result، Nested PCR technique investigated in this study، has significant sensitivity and accuracy; therefore، the molecular technique as a confirmatory method in detection of Brucella spp. seems necessary and inevitable for serological methods.