ابوالقاسم اسماعیلی

Levonorgestrel is considered as an effective compound in contraceptive pills and a lipophilic-hydrophobic substance with the ability to bind to suspended organic matters.In this study,the inclusion effects of levels of levonorgestrel at 0 (control), 0.7, 7, 70, 100 and 150 ng/g of diet and a control-solvent treatment were evaluated on growth indices of gourami (Trichopodus trichopterus) during 45 days.Females (n=210;initial weight and length of 4.6 g and 63.2 mm, respectively) were randomly distributed in 21 aquariums with density of ten fish per aquarium;and fed daily at the rate of 3% of body weight.Results showed that body height, condition factor and survival rate of females were not affected by levonorgestrel levels.Females fed with levonorgestrel showed significant differences in terms of final weight and length, increment percentage in weight and body length, weight and length specific growth rate compared to the control(and control-solvent treatment). Nevertheless, the growth indices and maximum abdominal diameter of females fed with 150 ng/g showed no significant differences with lower levels (up to 7 ng/g) (P>0.05). Based on the results, it can be concluded that the addition of levonorgestrel in the diet of gourami females led to developmental disorders, especially at a level higher than 7 ng/g. Due to the binding ability of levonorgestrel to organic substances and the possibility of accidental feeding of these particles in the natural diet of fish, growth disorders and changes in the population dynamic of this species and probably other groups of aquatic animals can be expected in natural environments.

Investing in health and safety has long been considered costly and inevitable, and the cost factor has always been a barrier to implementing safety and health interventions. Decision-makers in this area must demonstrate that such interventions are not only effective in achieving their goals but also have high economic efficiency. The present study aimed to determine the optimal model for economic evaluation in occupational safety and health investments.

The Analytic Hierarchy Process (AHP) method and Expert Choice® software were used to determine the optimal model. First, the OSH economic evaluation models/ tools were identified based on a review of previous studies. Then, considering 7 main criteria, the models were evaluated and the optimal model was introduced.

By literature search and screening of the findings, 20 models/tools were identified. then, using a decision matrix, modelschr(chr(chr(chr('39')39chr('39'))39chr(chr('39')39chr('39')))39chr(chr(chr('39')39chr('39'))39chr(chr('39')39chr('39')))) specifications were extracted and compared with each other. 4 models were identified for prioritization with AHP. The accuracy and precision with a final weight of 0.490 and the comprehensiveness with a final weight of 0.076 were the most and least important criteria, respectively. After prioritizing the alternatives over the target, the ROHSEI model, ToolKit method, Productivity Assessment tool, and Potential Method weighed 0.320, 0.281, 0.240, and 0.159, respectively.

HSE professionals can use ROHSEI as a tool for demonstrating the financial incentives for their goals, allocating limited resources, and improving the economic efficiency of their projects.

Anemia is a common complication of chemotherapy. In order to resolve this problem, multiple red blood cell transfusions are administered, leading to iron overload. Given the confirmation of positive correlation between the increased bone marrow iron stores and adverse response to the treatment in the previous study, the effect of iron on the proliferation of acute lymphoblastic leukemia (ALL) cell lines cell lines and its underlying mechanism were investigated in the current study.

Nalm6 and CCRF-CEM cell lines were selected as representatives of B-ALL and T-ALL Immunophenotypes and were treated with different concentrations of holo-transferrin (1 - 120 µM) and ferric ammonium citrate (FAC) (400 - 25000 µM). The cellular iron uptake was confirmed by AAS test. The cell proliferation and levels of intracellular free radicals (ROS), were evaluated by MTT and flow cytometry, respectively. Statistical analysis was performed using GraphPad prism software by one-way ANOVA test.

The effect of holo-transferrinon cell proliferation was not significant. However, FAC enhanced the proliferation rate of both ALL cell lines over 50%. Evidence showed that iron induced intracellular ROS, so that FAC in the concentration of 400 µM may induce the intracellular ROS over 50% (55.27 6 6.36% vs. 100%, p < 0.001).

Iron can support these cancerous cells by inducing ROS and augmenting leukemic cell proliferation. Therefore, the present study suggests that the volume of the injected blood in ALL patients should be minimized to prevent iron overload

Thyroid carcinoma is the most common endocrine cancer. There are several methods for determining the mutation of genes, one of them is random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR). This study aimed to compare DNA changes in the genome of thyroid hyperplasia, thyroid papillary carcinoma, and healthy tissues using RAPD-PCR. In this case-control study, 49 patients with papillary carcinoma and thyroid hyperplasia which had normal, hyperplasia, and papillary carcinoma tissues of the thyroid gland were randomly selected. Then, the primer pattern of the sequence 5'-AAGAGCCCGT-3' containing 10 pairs base was evaluated and compared between the tissues. The prevalence of 370-bp pair base band observed in normal, hyperplasia, and pylori thyroid carcinoma tissues was 6.1, 53.1, and 89.8 percent, respectively, and there was a significant difference between the three tissue types (P < 0.001). There was also a significant difference between normal tissue with hyperplasia and papillary carcinoma of the thyroid and between hyperplasia with thyroid carcinoma in this regard (P < 0.050). It seems that using RAPD-PCR is a suitable method for the detection and differentiation of normal, hyperplasia, and papillary thyroid carcinoma cells. Bat notice to the limitation of the study, more investigation in this field is recommended.

Deoxyribozymes are oligoribodeoxynucleotides that catalyze reactions such as cutting RNA and have diagnostic and therapeutic applications. Deoxyribozyme 10-23 includes a catalytic domain dependent on a fixed 15-nucleotic (mer) cation and two variable binding arms that cause the specificity of enzymes. Lactose operon is used in the white-blue screening process. This operon includes three polycistronic genes. In this study, a deoxyribozyme against α-peptide beta-galactosidase gene in the lactose operon was designed.
pGEM-T map was obtained from addgene server and α-peptide gene sequence was determined. Then, using expasy website proper protein frame in comparison with various reading frames was determined. In this step, whole sequence was reversed and mRNA sequence was achieved. Secondary structure with the lowest free energy was gained using mfold server. Considering the fact that 10-23 deoxyribozyme has cutting capability between a unpaired purine and pairs pyrimidine; an AC was selected in ribosome binding site in the untranslated region and then 9 open bases on either side of it was used as a binding arms. Investigation of the absence of similar sequences in host bacteria was performed by NCBI server. Finally, activity and binding of deoxyribozyme was predicted by the mfold server.
The results of this study showed that the designed deoxyribozyme had a relatively high Tm with two 9-nucleotide arms, which increased its effectiveness.
The results of this study can be used to control the expression of lacZ gene as a biomarker.

Vinegar is a popular condiment in the world that different materials and methods have been used to produce it. In Iran natural vinegar is also prepared mostly in a traditional way by using different fruits such as grapes and apples. Natural vinegar has beneficent properties and because of this, it is recommended to be used by traditional and Islamic medicine. Vinegar contains acetic acid bacteria, lactic acid bacteria and yeast. Acetic acid bacteria and yeasts are involved in the production of vinegar and lactic acid bacteria improve the flavor of vinegar. The aim of this study was isolation and identification of lactic acid bacteria especially Lactobacillus brevis from traditional vinegar.
After collecting a few traditional vinegars, the vinegar samples cultured for isolation of lactic acid bacteria on MRS broth and agar media contained nystatin as an anti-yeast antibiotic. Then some microbiological tests including catalase, gram staining and fermentation of carbohydrates were performed. Then, they were cultured at different temperatures, pH and different concentrations of salts. Finally, three isolates bacteria with biochemical properties of Lactobacillus brevis were evaluated by16 srDNA gene amplification.
Twelve lactobacilli were isolated from three vinegar samples. All isolated bacteria were catalase-negative and gram-positive. They could be able to grow at pH around 4.5 and 5.6, and at 2, 4 and 5.6% of salt concentrations. Most of the bacteria grew at 15oC, whereas one isolated grew at 45oC. Sequencing and Blast results showed that the three strains are Lactobacillus brevis.
Discussion and Lactobacillus brevis and Lactobacillus plantrum were found in traditional vinegars. Although isolation of Lactobacillus plantrum from vinegar was reported previously, as far as we could determine, it is for the first time that we could isolate Lactobacillus brevis from vinegar.

Gamma-aminobutyric acid (GABA) is a four-carbon non-protein amino acid used in the treatment of hypertension, diabetes, inflammation, and depression. GABA is synthesized by glutamic acid decarboxylase (GAD) enzyme in many organisms, including bacteria. Therefore, cloning of this enzyme is essential to the optimization of GABA production. This study aimed to clone and construct the expression vector of GAD gene from Lactobacillus plantarum PTCC 1058 bacterium.
In this experimental study, we investigated the morphological, biochemical, genetic and 16s rDNA sequencing of L. plantarum PTCC 1058 strain. Genomic DNA of the bacterium was isolated and amplified using the GAD gene via polymerase chain reaction (PCR). Afterwards, the gene was inserted into the pJET1.2/blunt cloning vector and subcloned in vector pET32a. Plasmid pET32a-gad expression vector was transformed in Escherichia coli BL21 strain, and protein expression was assessed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).
FINDINGS: Morphological, biochemical and genetic analyses of 16s rDNA sequencing indicated that the studied substrain was of the L. plantarum strain. In addition, results of nucleotide sequencing of the fragmented segment via PCR showed the presence of GAD gene. Results of colony PCR and SDS-PAGE analysis confirmed the accuracy of the cloning and gene expression of the recombinant Escherichia coli BL21 strain.
According to the results of this study, cloning of GAD gene from L. plantarum PTCC 1058 was successful. These cloned genes could grow rapidly in prokaryotic and eukaryotic systems and be used in cost-effective culture media and even non-recyclable waste.

Nowadays، recombinant products have been produced using transgenic animal technology. So far، several techniques have been used to produce transgenic organisms. The most recent of these techniques is SMGT that has been established based on the ability of the sperm to uptake exogenous DNA and transfer it to the oocyte during fertilization. SMGT can be an efficient method when coupled with complementary methods such as electroporation، lipofection، ICSI، REMI and other techniques. The mechanism of the action of SMGT is not well defined. However، it seems that entering exogenous DNA into the sperm genome is a non-random event. The sperm chromatin condenses defines the limited sites for DNA integration. SMGT accompanied by other methods such as electroporation، lipofection، ICSI، REMI and other techniques can be an efficient method، but each of these methods has different efficiency and effects in different species. It is now clear that the sperm has the ability to transfer DNA molecules to egg during fertilization. Recent advances have reported that the efficiency of SMGT is growing and it is a cheap، fast and efficient technique for producing transgenic animals.

Gamma-aminobutyric acid (GABA) is a non-protein amino acid that can be found broadly in all organisms. GABA is synthesized by glutamate decarboxylase (GAD). Therefore, GABA production depends on biochemical properties of this enzyme. GABA has several physiological functions such as hypotensive activity, insomnia, depression, as well as diuretic effects. Bacteria, fungi and yeast produce considerable amount of GABA. Among bacteria, acid lactic bacteria due to their physiological and safety properties have been considered. These bacteria are used in food industry and act as probiotics in digestive system. The aim of this study was to identify and introduce GABA producing acid lactic bacteria. In this paper using data banks such as PubMed and Google scholar and key words such as GABA producing acid lactic bacteria, isolation sources of them, factors that affect GABA production, acid lactic bacteria glutamate decarboxylase properties, glutamate decarboxylase gene cloning and regulation of them, the potential applications of GABA producing acid lactic bacteria and strain screenings have been reviewed. Data from variety of papers showed that production of GABA using acid lactic bacteria is safe and biocompatible. This may lead to GABA rich fermented products. Results of this study showed that natural GABA has significant effects on human health. Therefore it seems acid lactic bacteria are the most suitable sources for GABA production.

Multidrug resistance (MDR) is the main obstacle against treatment of acute lymphoblastic leukemia (ALL). ATP-binding cassette transporters function is mentioned as one of the most effective factors on MDR development. Though، there are many evidences on interference of ABCG2/BCRP، one of the outstanding members of this superfamly، in MDR occurrence، the expression effect of this gene on blasts of ALL patients is unknown yet. In this study، we used Real-Time PCR technique in order to investigate the relative expression of ABCG2/BCRP mRNA in 1-17 year old children with ALL. Peripheral or bone marrow blood samples from 28 new case leukemic and 15 control children were investigated with cooperation of Seyyedo-Shohada hospital in Isfahan،. Minimal Residual Disease (MRD) was evaluated as quality of patient response to chemotherapy. Profile of ABCG2/BCRP mRNA expression did not have any significant difference in new case ALL patients in comparison with control children. On the other hand، comparison of two groups of MRD+ and MRD- patients showed no difference in ABCG2/BCRP expression level. The level of ABCG2 expression was not associated with immunophenotype of ALL or known prognosis factors for this type of leukemia. Results of this study showed no effect of ABCG2/BCRP expression level on MDR development in ALL. Accordingly، clinical value of ABCG2/BCRP expression profile determination was rejected as the prognosis value for childhood ALL in our geographical area.

The aim of this study was investigation of growth rate, survival and activity of digestive enzymes (α-amylase and alkaline phosphatase) in fish larvae fed Artemia nauplii.The experiment was performed in 115-liter glass tanks containing 7000pieces of kutum larvae with an average initial weight of 5.1 mg (2 days after hatching).Samples for analysis of digestive enzyme activity were taken randomly at 2, 3, 5, 8, 10, 13, 17, 23, 27, 30.The results displayed that the stocking in high density to decrease growth efficiency but there was not considerable effect on survival rate larvae.In the present study was observed a significant increase in activity ofamylase from 23 to 30daysas the highest amylase activity was found at 30 days after hatching. The high content of glycogen and carbohydrates in live food may be to stimulate synthesis and secretion of amylase. An early peak of activity in alkaline phosphatase enzyme activity was observed at day 8 after hatching the indicating the maturation process in the intestines of kutum larvae.

Inflammation is the normal response of the body to injuries to remove dead cell debris and infections at injury sites and repair the tissue. However, excessive inflammation is harmful to the body because of its positive feedback and disruption of normal cells. Thus, recruitment of the regulatory mechanisms for modulating the inflammation is crucial for the body. Peroxisome Proliferator-Activated Receptors (PPARs) are ligand-activated transcription factors and members of the nuclear receptor super family. Among these superfamily members, PPARγ ia involved in the regulation of numerous cellular processes such as inflammation. Several activators of PPARγ have been known to exert possible roles in immune function through mechanisms such as transactivation of genes that suppress the inflammation, trans-repression of inflammatory genes, interaction with other receptors that lead to effective suppression of the inflammatory agents. Here in this review article, we describe the molecular events which are triggered by these molecules in elaboration of inflammation process.

Peroxisome proliferator-activated receptors (PPARs) are nuclear hormone receptors comprising three different isotypes termed β/δ،α and γ. They act at the transcriptional level. PPARγ is expressed primarily in adipose tissue primarily in adipose tissue and is considered as an adipogenic factor which regulates the expression of genes associated with lipid metabolism. The ligandactivated nuclear receptor peroxisome proliferator-activated receptor gamma (PPARγ) has been shown to regulate cell activation, differentiation, proliferation, and/or apoptosis. Alternative promoter regions within the PPARγ gene are responsible for the formation of three PPARγ isoforms. Occurrence of (Pro12Ala) polymorphism was found to be significantly associated with a change in lipid profile in obese carrier individuals. PPARγ is the target of the insulin sensitizing thiazplidinediones (TZDs), a class of currently used drugs in the treatment of type2 diabetes mellitus. PPARγ plays a major role in tissue differentiation. The repressive effects of PPARγ on tumor carcinoma cell lines have opened new hopes for therapeutic applications of this factor to inhibit the progress of cancer

HEK (human embryonic kidney) 293 cells as transient transfection systems are the most useful expression method to study physiological and pharmacological properties of the GABAA receptor subunits. HEK properties are similar to the native expressed channels. In comparison to the other common artificial expression systems، HEK cells have a similar mammalian intracellular environment. Since some mRNAs such as GABAA 3 are significantly found in HEK 293 cells، caution must be taken in interpreting the results of studies of the properties of GABAA receptors expressed in HEK 293 cells. In this study we examined the endogenous mRNA content of GABAA receptor subunits Alpha1 and Alpha2 and Gama1 and Gama2 in HEK 293 cells using traditional PCR and real time PCR. Total cellular RNA was extracted from HEK 293 cells and reverse transcribed to cDNA. PCR and real time PCR reactions were performed in a 25 l volume containing specific forward and reverse primers for GABAAR subunits with other required reagents. Reactions were carried out using a thermocycler. The amplification products were analyzed on 2 % agarose gels stained with ethidium bromide. In this study، none of GABAAR subunits in HEK cells were detected. The results show that HEK 293 cell line can be used as an expression system for the analysis of GABAA receptor recombinants.

Alternaria brassicae is one of the most important seed borne pathogens and causal agents of canola leaf spot. In pathogenicity process of A. brassicae, AbreAtr1 genes correspond to pathogenicity cluster NRPS-ABC transporter and have been introduced as an encoding of pathogenicity factor. The presence of these proteins in organisms prevents the accumulation of toxic compounds caused by them, and protects them against toxins, thus enhancing the growth of pathogen. The role of this gene in growth of A. brassicae and amount of pathogenicity of this fungus has not yet been considered. In order to study this role, the amount of AbreAtr1 transcription in six isolates of A. brassicae was compared by Real time PCR method. The daily growth rate in each isolate in PDA medium and the amount of pathogeniciy was also studied. There was a significant difference among isolates at 1% level in the growth rate and amount of pathogenicity. Also, there was a significant difference among isolates in the amount of AbreAtr1 transcription. The role of AbreAtr1 in protective mechanism of A. brassicae against the produced phytotoxins, the correlation between the growth rate, amount of pathogenicity and transcription pattern of AbreAtr1 were determined in each isolate. There was a positive relation between growth rate, amount of pathogenicity and transcription pattern of AbreAtr 1 at 1% level. It is thought that growth and pathogenicity of A. brassicae is affected by the ABC transporter encoded by the AbreAtr1. Over-expression of AbreAtr1 can lead to reduce toxic effect of secondary metabolites in this pathogen, thus giving more pathogenicity in the isolate.

Brain memory storage and information processing occurs through changes in synaptic connections between neurons or synaptic plasticity. In experimental models of synaptic plasticity such as LTP, stabilization of these changes requires de novo protein synthesis. Immediately after stimulations that induce LTP or behavioral experiments which contribute to Long Term Memory (LTM) formation, Immediate-Early Genes are induced in hippocampal neurons; among neuronal activity dependent IEGs, Arc is of particular importance. Arc is robustly induced by plasticity-producing stimulation and is specifically targeted to stimulated synaptic areas. It also plays a critical role in the maintenance of long term potentiation and in memory consolidation processes. Inhibition of Arc protein expression in the rat hippocampus, through ODNs infusion, impairs maintenance of LTP and consolidation of LTM. Thus, Arc appears to play a fundamental role in stabilization of activity-dependent hippocampal plasticity. In this paper, the role of Arc in memory formation is discussed.

Hearing loss is the most frequent sensory disorder affecting 1 in 500 neonates with more than 50% of inherited cases. This trait is a very heterogeneous disorder and happens due to genetic or environmental causes or both. More than 46 genes may be involved in non-syndromic hearing loss. Recently, DFNB59 gene has been shown to cause deafness in some Iranian populations. The aim of this study was to determine the role of DFNB59 gene mutations causing deafness in a group of 130 deaf pupils in Fars province.

This descriptive-laboratory based study investigated the frequency of DFNB59 gene mutations using PCR-SSCP/HA strategy.

Two different DFNB59 polymorphism including 874G>A and 793C>G were found in 1 and 9 of 130 patients studied respectively. However, no DFNB59 mutation was identified.

The results of this study shows that the association of DFNB59 mutations with deafness in Fars province is very low.

Adeno-associated virus (AAV) is a small virus encapsidates a single-stranded DNA genome. AAV have rapidly gained popularity as a vector in genetic engineering and gene therapy applications, due to its lack of pathogencity, wide range of infectivity, and ability to integrate into human genome (19q13.4). Recombinant AAV vectors have been used for treatment of a variety of diseases such as hemophilia, cystic fibrosis, and Parkinson. A challenge with wide application of this vector is the relatively small packaging size of the viral genome to enable the delivery of a large transgene. Therefore it is essential making changes in it to overcome this problem. In this review we will introduce with this virus and the new finding in reginal of gene therapy with this virus.

The aim of this study was to clone PPARγ1 cDNA in an appropriate mammalian expression vector, with a chimeric cDNA form, encompassing PPARγ with enhanced green fluorescent protein (EGFP) cDNA. This recombinant plasmid will be used for further analyses to investigate the molecular mechanism of PPARγ1 for neural differentiation process. Moreover, the nuclear localization of the PPARγ1 protein linked to EGFP marker was chased by using transient transfection of a constructed plasmid into bovine fibroblast cells. Total RNA was extracted from the fatty tissue of an adult mouse. Using specific pair primers, PPARγ1 cDNA was synthesized and amplified to produce the entire length of ORF. RT-PCR products containing PPARγ1 cDNA were treated by enzymatic digestion and inserted into the pEGFP-C1 downstream from EGFP cDNA. The constructed vector was used for transformation into bacterial competent cells. Positive colonies which showed inserted PPARγ1 cDNA were selected for plasmid preparations and additional analysis was performed to ensure that PPARγ1 cDNA was inserted properly. Finally, to confirm the intracellular localization of EGFP-PPARγ1, bovine fibroblast cells were transfected with the recombinant plasmid. Our results from enzymatic digestion and sequencing confirmed, as expected, that PPARγ1 cDNA was amplified and cloned correctly. This cDNA gene encompassed 1428 bp. The related product was entered into the nucleus of bovine fibroblasts after transfection of its cDNA. PPARγ1 cDNA was cloned and sorted into nuclear compartments of bovine fibroblast cells upon transfection.

The main aim of this research is to study nuclear localization of mouse PPARγ1 cDNA into pEGFP-C1. To investigate the nuclear localization of PPARγ1 protein linked to EGFP marker gene into bovine fibroblast and analysis of intracellular green fluorescency Mouse PPARγ1 cDNA in a mammalian expression vector (pEGFP-C1) in a chimeric cDNA type, encompassing PPARγ1 with EGFP cDNA. EGFP marker was used for monitoring. To confirm the intracellular localization of EGFP- PPARγ1, bovine fibroblast cells were transfected with the 2.4 µg constructed plasmid and 6 µl Lipofectamine 2000. The related product was entered into the nucleus of bovine fibroblasts after transfection of its cDNA. As expected, chimeric cDNA of EGFP-PPARγ1 correctly expressed and related protein was dominantly entered to the nuclei of the cells. Thus the recombinant plasmid could be applicable for further studies to unravel the further aspects of PPARγ1 function. Moreover fusion of EGFP with PPARγ1 does not hamper the nuclear targeting of PPARγ1.

Peroxisome proliferator-activated receptors (PPARs) belong to nuclear receptor family, which act as lipid sensors to modulate gene expression and control cellular fate. PPARs are implicated in different cellular processes such as cellular differentiation, apoptosis, development, immune responses and energy homeostasis. PPARs compose a family of three nuclear receptors that they are termed PPARα, PPARβδ and PPARγ. These receptors demonstrate differential ligand specificities and cellular expression patterns. These PPAR ligands are used to treat type II diabetes mellitus, hypertriglyceridemia and inflammation. Recently, these receptors have been supposed as new targets for the treatment of CNS inflammatory disorders. Throughout this review, we focus on the molecular structure of PPARs, posttranslational modifications that regulate function of these receptors and then molecular mechanisms through which PPARs regulate transcription with particular emphasis on the latest results on corepressors and coactivators actions. Finally, we introduce each PPAR isotype, cellular functions of them and their ligands.