mortaza bonyadi

Physiological studies confirm improvement of memory by Olibanum, a resin from Boswellia species, while little is known about the molecular mechanism by which it affects memory performance. Two master transcription factors, CREB-1 and CREB-2, regulate downstream memory-related genes expression, leading to the long-term memory potentiation. This study addresses the effects of Beta-boswellic acid (β-BA), the main ingredient of Olibanum, and ethanolic extract of the resin from Boswellia serrata on the expression of CREB-1 and CREB-2 genes in B65 cell line. B65 cells were treated with β-BA) or ethanolic extract of Olibanum in different dose and time intervals and the cell viability/toxicity was measured by MTT assay. Total RNA was extracted from the treated and untreated control cells and cDNA was synthesized. The expression levels of CREB-1 and CREB-2 genes were quantified by Real-time PCR. MTT assays revealed 50% inhibitory concentrations of 42.05, 29.63 and 21.78 μg/ml for ethanolic extract of Olibanum and 89.54, 44.05 and 21.12 µM for β-BA at 24, 48 and 72h time intervals respectively. Both β-BA and ethanolic extract of Olibanum altered CREB-1 and CREB-2 genes expression levels in time-dependent but not in dose-dependent way. However, β-BA showed stronger and more stable effects. The expression levels of the both genes followed an alternate upregulation and downregulation pattern, but in opposite directions, in response to the both solutions with the progress of time. These results suggest that Olibanum possibly improves memory performance, at least partially, by regulating the levels of CREB-1 and CREB-2 transcription factors via positive/negative-feedback loops.

In the past, olibanum was used as an incense and perfume and a remedy for treating various diseases. Now, it is confirmed that olibanum improves memory and has a role in treatment and prevention of Alzheimer disease. CaMKIIα gene is known as molecular memory because of its role in cellular processes associated with memory. The aim of this research was to study the effect of ethanolic extract of olibanum on the expression of CaMKIIα in B65 and PC12 cells. To determine the effect of olibanum on cell viability, MTT assay was done and cells were treated with the extract in different concentrations and times. To study the gene expression, the cells were treated by two concentrations of the extract at four time points. Then, RNA was extracted, cDNA was synthesized, and the expression of CaMKIIα was quantified by qPCR. The qPCR data revealed that olibanum alternately changed the expression of CaMKIIα in a reducing and increasing pattern over time in B65 cells. However, the extract could not induce the expression of this gene in PC12 cells. According to the role of CaMKIIα as molecular memory and the positive effect of olibanum in memory improvement, current results could be helpful in understanding the molecular mechanisms by which olibanum affects memory performance.

Long-term memory depends on protein synthesis. The product of Camkiv gene promotes memory via activating its proteins. The treatment of laboratory animals by Olibanum leads to memory improvement and the recovery of Alzheimer. Therefore, the aim of this study is the evaluation of Olibanum ethanolic extract on the Camkiv expression in PC12 cells. Olibanum toxicity on the cell viability was investigated by MTT test. Cells were treated with concentrations 10,25,40,55,70 and 85 μg/ml of extract in time intervals 12,24,48 and 72 hours and their absorption rate was measured. Then, cells were treated by concentrations 2 and 20 μg/ml of extract in mentioned times. Extracted RNA was converted into cDNA and real-time PCR performed. Cell death was raised by increasing time and concentration of extract treatment. IC50 values were obtained as 71.01, 52.95, 21.05 and 13.85 μg/ml in 12, 24, 48 and 72 hours of treatment, respectively. Besides, concentrations 2 and 20 μg/ml significantly increased Camkiv expression following 24 hour treatment. The maximum expression of Camkiv was observed in 48 hour treatment. The effect of Olibanum on gene upregulation was stable until 72 hours. The Olibanum ethanolic extract can remarkably upregulate Camkiv expression for a long time. These results are consistent with the previous studies indicating the effect of Olibanum on upregulation of Bdnf, Camkiv -downstream gene. However, regarding the existence of the two-direction pathway in the expression regulation of Bdnf and Camkiv , comprehensive studies are required to determine exact mechanism of Olibanum function in the brain.

Familial Mediterranean Fever (FMF) is the most common inherited autoinflammatory disease. Kidney involvement in FMF is usually attributed to secondary amyloidosis. Non-amyloid glomerular involvement has also been reported.
We suppose that heterozygous mutation of Mediterranean fever (MEFV) gene could be the underlying cause in some cases of mesangial proliferative glomerulonephritis (MePGN) in FMF endemic area.
This prospective study was done between 2013 and 2015 in NorthWest of Iran among the Azari-Turkish population. A panel of MEFV gene including M680I, R761H, M694V, R408Q, E148Q, A744S, F479L, P369S, V726A, M694I, and E167D were studied in a group of patients with idiopathic MePGN. Clinical characteristics and therapeutic responses were compared between those with and without a mutation. A total of 39 idiopathic MePGN patients and 156 healthy subjects were studied.
Heterozygote mutations of MEFV gene were detected in 11/39 (28.2%) of MePGN patients and 46/156 (17.3%) of controls. Clinical response regarding 24 hours urine protein excretion was significant in mutation-negative patients after 6 months of follow-up.
This study shows a possible underlying role of heterozygous MEFV gene mutation in the clinical course of some case of idiopathic MePGN, particularly in FMF endemic population.

The most important effects of Frankincense on memory are included prevention and relative treatment of Alzheimer disease, as well as memory enhancement of off springs of the rats that had received Frankincense during their pregnancy and lactation period. Considerably, Camk4, one of the important memory genes, induces downstream memory genes expression through phosphorylation and activation of transcription factors. Here, we aimed to study the effects of aqueous extract of Frankincense on the Camk4 gene expression in PC12 and B65 cell lines.
The effect of extract on the cell viability was evaluated by cell treatment in two time intervals with six concentrations of extract and 50% inhibition concentration was obtained. In this way, for gene expression studies, cells were treated by two concentrations of extract in two time points. RNA was extracted and converted to cDNA and qPCR was performed to investigate the expression of Camk4 gene.
The aqueous extract of Frankincense decreased the cell viability in a time and concentration dependent pattern. However, the extract was more toxic for B65 cell line than PC12. Also, it significantly increased the expression of Camk4 gene in the cells. Nevertheless, the increase was dependent to the extract concentration and the cell type. The low concentration of the extract was more effective to the expression of Camk4 gene than the high concentration as well as to its expression in the PC12 cell line than B65.
This study showed that Frankincense could regulate the expression of Camk4 gene in a concentration dependent pattern. However, further studies are needed to identify the mechanisms of Frankincense action in different cells.

Age-related macular degeneration (AMD) is the leading cause of blindness in the developed world and is characterized by progressive degeneration of the retinal pigment epithelium and secondary photoreceptor loss, resulting in visual loss. Etiological research suggests that age related macular degeneration is a complex disease, caused by the interactions of several genetic and environmental factors. Polymorphisms in genes encoding the alternative complement pathway, complement factor I (CFI), are associated with the risk for age related macular degeneration. The purpose of this investigation was studying of complement factor I p.Gly119Arg (C.355G>A) polymorphism with age related macular degeneration in the population living in Tehran, Iran.
This case-control study was conducted at Tabriz University from June 2015 to June 2016. In this study the association of p.Gly119Arg polymorphism in complement factor I gene was investigated in 150 patients suffering from age-related macular degeneration and 150 healthy age, sex and ethnicity matched unrelated people as control group. Both of the case and control groups were originated from the population living in Tehran. Genotypes of both groups were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and data was analyzed the Chi-square test in 2x2.Contingency table.
Investigation of the association of p.Gly119Arg polymorphism in complement factor I gene with age related macular degeneration showed that there are statistically significant differences between patients and controls in genotype and allele frequencies of this polymorphism (P=0.005 and OR=6.68 in TT, P=0.04 and OR=0.61 in CC, P=0.03 and OR=1.76 in T, P=0.04 and OR=0.56 in C). Therefore CC, TT genotypes and C, T alleles were significantly associated with age related macular degeneration.
This study showed a significant association between this polymorphism p.Gly119Arg (C.355G>A) complement factor I gene and age related macular degeneration disease in the population living in Tehran (P

To investigate the association of CFI p.Gly119Arg polymorphism with Age-related macular degeneration (AMD).
In this case-control study, the association of p.Gly119Arg polymorphism in CFI gene was investigated in 65 patients suffering from AMD and150 healthy age, sex and ethnicity matched unrelated people as control group. Both of the case and control groups were originated from Northwest of Iran. Genotypes of both groups were determined by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP).
Investigation of the association of p.Gly119Arg polymorphism in CFI gene with AMD showed that there are no statistically significant differences between patients and controls in genotype and allele frequencies of this polymorphism (P˃0/05).
Our result show that p.Gly119Arg polymorphism of the CFI gene is unrelated to the susceptibility to AMD in Northwest of Iran.

Age-related macular degeneration (AMD) is the most common cause of irreversible vision loss and debilitating disease in old age, which involves the central retina/macula among elderly patients. The genetic and environmental factors have important role in this multifactorial disease. Oxidative stress and DNA damages would have important impact on the onset and progression of AMD.  In this study, the possible association of c.-31A>G (rs3087404) polymorphism in the promoter region of SMUG1 gene with AMD disease was investigated. Fifty five AMD patients and 130 healthy age-, gender- and ethnicity-matched unrelated people as control group were genotyped by restriction fragment length polymorphism PCR (RFLP-PCR). Both groups were from Northwest of Iran (Tabriz).  Statistical analysis showed a significant association of AG genotype of this polymorphism with AMD. These results suggest a possible protective effect of this genotype for AMD disease (P=0.02, OR=0.574) among patients from Northwest of Iran. This genotype was observed more frequently in controls compared to the patients (59.23% v s 45.45%).

Coexistence of Familial Mediterranean Fever (FMF) with various systemic vasculitides, such as Henoch Schonlein Purpura (HSP) and other inflammatory disorders has been reported and MEFV gene has been suggested to play a significant role in the pathogenesis of this association. In this study, the rare MEFV mutations in patients with HSP from north west of country and its association with clinical symptoms of disease were evaluated.
Material and Forty unrelated patients were referred by specialists to the Molecular Medical Genetic Center of Tabriz. Clinical diagnosis of HSP was made according to published criteria. The control group consists of 200 ethnically matched persons apparently healthy without any kind of inflammatory diseases. Screening for the 3 mutations; R761H, P396S and R408Q were performed by using amplification refractory mutation system polymerase chain reaction (ARMS-PCR) and polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). X2 test and Fisher's exact test were used to statistical analysis.
Of 40 patients studied, 37 (92.5%) showed without mutation, while 3 (7.5%) had MEFV mutation that three of them were compound heterozygous for the P396S/R408Q mutations. There was a statistically significant difference between the patient group and healthy individuals regarding P396S and R408Q mutations (p = 0.0043). Findings suggest that P369S and R408Q mutations always together occurred and not only contribute to the susceptibility to HSP, also associated with clinical symptom of fever.
Our results suggest that some MEFV mutations could be a contributory genetic factor to HSP in the north west of the country.

Immunological factors are important in pregnancy loss because of the interaction between mother and fetus. T-regulatory cells as the component of humeral immune response play important role in the fetu-maternal interface. One of the regulatory mechanisms for these cells is mediated by antigen independent co-stimulatory signals and interaction of Cytotoxic T-Lymphocyte Antigen 4 (B7/CTLA-4) is one of these signals. The CTLA-4 which down regulates the activation and proliferation of T-cells occurs in a competitive interaction with CD28 to bind to B7. The aim of this study was to find out the relationship of CTLA-4  gene with Recurrent Miscarriage in a group of Iranian women.
In the present study, 60 women with the history of two or more pregnancy loss were selected and considered as the case group. A group of women (n=60) with at least two live births without any previous history of pregnancy loss and autoimmune diseases were taken as control group. Genomic DNA was extracted from whole blood using standard protocols. The CTLA-4 A/G were detected using polymerase chain reaction-restriction fragment length polymorphisms assay.
The results showed that CTLA-4 A/G polymorphisms were not significantly different in women with the history of two or more pregnancy loss compared to normal individuals. The frequency of G-allele polymorphism was 39.16% and 35.83% in patients and controls respectively.
The data presented may suggest that the CTLA-4 is not associated with recurrent miscarriage in an Iranian population in Northwest region.

Henoch-Schönlein purpura (HSP) is an lgA mediated small vessel systemic vasculitis disease in children. The etiology and pathogenesis of HSP disease remain unknown. However, environmental and genetic risk factors could play important roles in susceptibility to HSP disease. In this study we investigated the association of 5՛-untranslated region polymorphism (-634G/C) of VEGF gene with HSP among Iranian Azeri Turkish population.
Thirty unrelated Iranian Azeri Turkish children with HSP and fifty healthy unrelated subjects without HSP and other inflammatory diseases were enrolled in this population. -634G/C polymorphism of VEGF gene was genotyped by polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) technique.
The distribution of CC genotype in VEGF -634G/C polymorphism statistically showed a significant difference in HSP patients in compare to that of control group (P= 0.009).
The CC genotype of VEGF -634G/C polymorphism could be associated with susceptibility to HSP disease in Iranian Azeri Turkish ethnic group.

Recurrent miscarriage (RM) is a pathologic condition that occurs in 1- 5% of couples trying to conceive and has destructive psychological effects on family life of affected couples. RM is considered as a multi-factorial disease and nearly half of RM cases cannot currently be explained clinically. Several studies have indicated that genes polymorphisms could be factors for causing susceptibility of RM with unknown etiology. GSTO1 protein has a crucial role in detoxification metabolism and it is believed that GSTO1 gene polymorphism could be involved in RM. This study aimed to investigate the possible association of GSTO1 gene polymorphism [E155del (rs 11509437)] with recurrent miscarriage from northwest of Iran was studied. In this case-control study, fifty-five patients with a history of RM and fifty-five referent women, who had at least two live births without other pregnancy complications, were included. Glutathione S- transferase Omega 1 (GSTO1) gene polymorphism (i.e. E155del) was screened using polymerase chain reaction with confronting two-pair primers (PCR-CTPP). A total of 55 patients were studied and 1.82% (1) patient for del155/del155 genotype of GSTO1 gene were homozygote (p=0.17). Allelic frequency of del155 between patients and the control groups were 9.09% (10) and 2.72% (3), respectively (p=0.023). The result of this study shows association of del155 allele of GSTO1 gene with susceptibility to develop recurrent miscarriage b ut there is no significant association between del155/del155 genotype and recurrent miscarriage in Northwest of Iran.

Age-related macular degeneration (AMD) is the most common cause of irreversible visual impairment among elderly population worldwide. Besides age and smoking, genetic variants from several gene loci have been associated with this condition. Several polymorphisms have been implicated in the pathogenesis of this disease. rs2230199 polymorphism in C3 gene that encodes the major protein of the complement system is one of the candidate genes involved in this disease. The present research examined the role of R102G polymorphism in developing of AMD. In this study, the association of rs2230199 polymorphism in C3 gene was investigated in 51 patients from Gonabad region suffering from dry AMD with Geogrophic atrophy by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). Some cases were chosen at random for confirmation of the PCR-RFLP results by direct sequencing. Also, 72 healthy age, sex matched unrelated people were included in this case-control study as controls which were selected from the same population. Investigation of the association of R102G polymorphism in C3 gene with AMD showed that there are significant differences between patients and controls in genotype and allele frequencies of this polymorphism (P= 0.000). rs2230199 polymorphism in C3 was associated with increased risk for AMD

Cystic fibrosis (CF), a life-limiting autosomal recessive disorder, is considered a monogenic disease that is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. According to several studies, mutation analysis of the cystic fibrosis transmembrane conductance regulator (CFTR) gene alone is insufficient to predict the phenotypic manifestations observed in cystic fibrosis (CF) patients. In addition, some patients with a milder CF phenotype do not carry any pathogenic mutation. Tumor Necrosis Factor-alpha (TNF-α) contributes to the pathophysiology of CF by causing cachexia. There is a reverse association between TNF-α concentration in patient''s sputum and their pulmonary function. To assess the effect of non-CFTR genes on the clinical phenotype of CF, two polymorphic sites (-1031T/C and -308G/A) of the TNF-α gene, as a modifier, were studied.Patients and Focusing on the lung and gastrointestinal involvement as well as the poor growth, we first investigated the role of TNF-α gene in the clinical manifestation of CF. Furthermore, based on the hypothesis that the cumulative effect of specific alleles of multiple CF modifier genes, such as TNF-α, may create the final phenotype, we also investigated the potential role of TNF-α in non-classic CF patients without a known pathogenic mutation. In all, 80 CF patients and 157 healthy control subjects of Azeri Turkish ethnicity were studied by the PCR–RFLP method. The chi-square test with Yates'' correction and Fisher''s exact test were used for statistical analysis. The allele and genotype distribution of the investigated polymorphisms, and their associated haplotypes were similar in all groups. There was no evidence that supported the association of TNF-α gene polymorphisms with non-classic CF disease or the clinical presentation of classic CF.

Henoch–Schönlein purpura (HSP) is an autoinflammatory disease and systemic small vessel vasculitis that more frequently occurs in children. It is characterized by skin lesion such as Petechia and purpura, gasterointestinal involvement including abdominal pain and gastrointestinal bleeding and arthiritis. Studies have shown that HSP could be due to different infections and genetic factors. Based on the fact that cytokines have important role in this inflammatory disease, CCL2- a chemokine encoding gene- is studied in HSP patients from northwestern Iran. To achieve this goal molecular analysis of polymorphism located in promoter region of this gene is performed. This study was conducted on 40 HSP patients who were referred by immunology and allergy specialists to Genetic Center of Tabriz and after blood sampling and DNA extraction, molecular analysis with PCR-RFLP method was performed to genotype polymorphic region of promoter region of this gene. The results were compared to that of 50 ethnic-sex matched control healthy people. Statistical analysis shows significant association of this polymorphism with development of Henoch–Schönlein purpura disease in this cohort. The frequency of TT and TC genotypes and T allele of CCL2 -C2518T gene polymorphism were significantly higher in HSP patients. CCL2 C-2518T gene polymorphisms were associated with susceptibility to HSP in Northwestern Iran.

Inflammatory bowel disease (IBD) is a chronic, relapsing, inflammatory disorder of the gastrointestinal tract that includes two entities, Crohn’s disease (CD) and ulcerative colitis (UC). As with other complex diseases, both genetic susceptibility and environmental factors play role in the pathogenesis of these diseases.The tumor necrosis factor α (TNF-α) gene is located in the IBD3 region on chromosome 6p21 which is a good functional candidate for involvement in susceptibility to IBD. In addition, the promoter region of TNF-α contains various polymorphisms that have shown a significant association with IBD. In this case control study we investigated the TNF-α -857 polymorphism in 109 patients (89 UC and 16 CD) who suffered from IBD and 100 healthy age, sex and ethnicity matched adults selected from the same population, as the control group. The polymorphism was checked by amplification refractory system (ARMS) and polymerase chain reaction (PCR). Investigation of the association of TNF-α -857 gene promoter polymorphism with both types of IBD showed no significant difference in genotype and allele frequencies of this polymorphism between UC patients and controls. However, a possible association of TNF-α -857 polymorphism (p=0.03) was identified with CD. TNF-α -857 polymorphism may have a role in the development of CD in the Iranian Azeri Turkish population.

Chromosome Y is a sex-determinant chromosome in human. Microsatellites located on this chromosome are being applied for paternity test, sex determinations in neonates and DNA finger-printing. The application of these markers in most cases depends on the polymorphic situation of them. The aim of this study was to study some of Polymorphic these markers located on Y chromosome among male population from Eastern-Azerbaijan. Fifty unrelated males and several females as a negative control were randomly selected from the interested population. DNA from whole blood cells were extracted and all the individuals were studied by three pairs microsatellites (DYS438, DYS390, DYS385) selected from the chromosome Y. PCR products were electrophoresed on ploy-acrylamid gel (SSR-PCR technique) and bands were analyzed. None of the studied markers were amplified in female controls indicating that these markers are male specific and they do not have a copy on autosomal or X chromosomes. DYS385, a poly-allelic marker, showed the highest percentage of polymorphism (95%) in this population. Markers DYS435 and DYS390 showed relatively less polymorphism respectively. This result indicates that in the population of East Azerbaijan DYS385 is one of the best markers for paternity test and DNA finger printing. In case of combination use of this marker with other two markers, the efficiency of the result would be highly increased