Evaluation of rs1982073 polymorphism of transforming growth factor-ß1 in glioblastoma.

BACKGROUND: Glioblastoma (GBM) is the most common and invasive form of primary malignant brain tumors, with a survival rate of about 1 year. Transforming growth factor-ß1 (TGF-ß1) plays a very important role in tissue homeostasis and cancers. It seems that polymorphism of T29C (L10P, rs1982073, or rs1800470), which has been studied in various cancers such as breast and colon, creates the significant differences plays an important role in GBM prognosis and treatment. In this study, we evaluated the effect of T29C (rs1982073) polymorphism of TGF-ß1 gene in GBM. MATERIALS AND METHODS: This study was conducted on 100 cases of GBM including 47 paraffin-embedded brain tissue samples and 53 blood samples from another 53 GBM patients, who was under therapy, and 150 were controls. The TGF-ß rs1982073 single-nucleotide polymorphism (SNP) was identified by the NCBI and genotyping was performed by high-resolution melt (HRM) assay. Melt curves from HRM which suspected to SNP were selected and subjected to direct sequencing. Finally, the collected data were entered into the SPSS software (Version. 20) and mean ± standard deviation or n (%) was used to show the data. RESULTS: The mean age in GBM group was 51.63 ± 13.27 years. Accordingly, the two groups were matched in terms of age and gender (P > 0.05). The frequency of GG genotype was significantly higher in GBM patients. In contrast, although the frequency of AG genotype was higher in GBM group, it was not statistically significant. Furthermore, the presence of G allele was significantly more frequent than A allele in GBM patients. CONCLUSION: Findings of the present study supports that the Pro10Leu, rs1982073, or rs1800470 SNP in TGF-ß1 is found to be expressed significantly more in GBM patients as it was found in breast cancer.

Evaluation of rs1957106 Polymorphism of NF-κBI in Glioblastoma Multiforme in Isfahan, Iran.

BACKGROUND: The kB family of nuclear factor (NF-κB) is a series of transcription factors that plays a key role in regulation of immunity, cell growth, and apoptosis and is considered as the main downstream component of epidermal growth factor receptor for which there are evidence of excessive activity in most cases of glioblastoma multiform (GBM). Thus, the current information has gained evidence on NF-κBIA tumor suppressor role in GMB. SNP rs1957106 was diagnosed as a new polymorphism which affected the expression of NF-κBI and causes activation of NF-κB in GBM patients. MATERIALS AND METHODS: This study was conducted on 100 cases of GBM including 47 paraffin-embedded brain tissue samples and 53 blood samples from another 53 GBM patients and 150 controls. The NF-κBI rs1957106 SNP was identified by the NCBI, and genotyping was performed by high-resolution melt (HRM) assay. Melt curves from HRM which suspected to single-nucleotide polymorphism (SNP) were selected and subjected to direct sequencing. RESULTS: The distribution of allele A of NF-κß gene in patients with GBM with 31% was not significantly different from healthy participants (27.3%) (P = 0.375). Furthermore, the distribution of AG and GG genotypes in comparison with AA genotypes did not show a significant correlation with GBM incidence (P > 0.05). CONCLUSION: Findings of the present study provide evidence that the rs1957106 SNP in NF-κBIA is found more in GBM patients, but it was not statistically significant. As there are conflicting studies showing significant higher rate of this SNP in GBM, further study is suggested.

Paraoxonase 1 activities and its gene promoter single nucleotide polymorphisms (-108, -126, and -162) in diabetes mellitus.

BACKGROUND: Paraoxonase 1 (PON1) enzyme is known enzyme with, aryl esterase, phosphatase, peroxidase, and lactonase activities. According to some studies, the activity of PON1 enzyme is decreased in type 2 diabetic patients. We analyzed the enzyme activity and its single nucleotide polymorphisms (SNPs) distribution on promoter regions (-108, -126, and -162) in type 2 diabetic patients compared with non-diabetic individuals to reveal the likely relationship between PON1 activity and its gene promoter polymorphisms. METHODS: On the whole, 98 diabetic and 104 non-diabetic individuals were examined in this study. The enzyme activity and the genotypes were studied using spectrophotometry, real-time PCR-HRM, and sequencing techniques, respectively. RESULTS: There was no meaningful difference in enzyme activity between two under-studied groups (P.V = 0.671). Moreover, no meaningful difference was also seen between two groups in terms of the frequency of polymorphism -108 (P.V = 0.277). The frequencies of SNPs -126 and -162, however, showed a meaningful difference between two groups (P.V = 0.000 and P.V = 0.017, respectively). CONCLUSIONS: We indicated PON1 activity could be similar in DM-2 patients and non-DM-2 individuals. The significant role of SNP -108 in PON1 activity in DM-2 patients compared with non-DM-2 individuals was confirmed in the study too. On the other hand, the role of -162 and -126 SNPs in causing diabetes cannot be easily overlook because of a meaningful difference of their distribution in understudied groups. However, they may be attributed to DM-2-associated genes.

Relationship of lipid regulatory gene polymorphisms and dyslipidemia in a pediatric population: the CASPIAN III study.

OBJECTIVE: In this study, we aimed to assess the association between four variants in three genes whose association has been reported in adults but not in children. We evaluated the relationship of the GCKR (rs780094), GCKR (rs1260333), FADS (rs174547), and MLXIPL (rs3812316) polymorphisms with serum lipid levels in Iranian children. DESIGN: This cross-sectional study was conducted in a subpopulation of the CASPIAN III study. During this study, 550 frozen whole blood samples were selected randomly. Using the recorded information of selected cases, those with and without abnormal lipid levels were determined. Allelic and genotypic frequencies of GCKR (rs780094), GCKR (rs1260333), MLXIPL (rs3812316), and FADS (rs174547) polymorphisms were determined and compared in dyslipidemic and normal children. The association between the studied polymorphisms and lipid profiles was determined using logistic regression analysis. RESULTS: Prevalence of hypercholesterolemia, hypertriglyceridemia, high low-density lipoprotein cholesterol (LDL-C), and low high-density lipoprotein cholesterol (HDL-C) were 24.9, 34.5, 19.0, and 40.7%, respectively. Significant correlations were found between GCKR (rs780094) and GCKR (rs1260333) polymorphisms and cholesterol and triglyceride levels, between FADS (rs174547) polymorphism and level of triglyceride, and also between MLXIPL (rs3812316) and levels of HDL-C. CONCLUSIONS: The results of this population-based study provide evidence for a relationship between lipid regulatory gene polymorphisms including GCKR (rs780094), GCKR (rs1260333), FADS (rs174547), and MLXIPL (rs3812316) with dyslipidemia in an Iranian population. These results could provide baseline information on as well as further insight into the genetic makeup of lipid profiles in Iranian children, which could be used for preventative strategies.

Lipid regulatory genes polymorphism in children with and without obesity and cardiometabolic risk factors: The CASPIAN-III study.

BACKGROUND: Genetically, predisposed children are considered as at-risk individuals for cardiovascular disease. In this study, we aimed to compare the frequency of four-lipid regulatory polymorphism in obese and normal-weight children with and without cardiometabolic risk factors. MATERIALS AND METHODS: In this nested case-control study, 600 samples of four groups of participants consisted of those with normal weight with and without cardiometabolic risk factors and obese with and without cardiometabolic risk factors. Allelic and genotypic frequencies of GCKR (rs780094), GCKR (rs1260333), MLXIPL (rs3812316), and FADS (rs174547) polymorphisms were compared in the four studied groups. RESULTS: Data of 528 samples were complete and included in this study. The mean (standard deviation) age of participants was 15.01 (2.21) years. Frequency of tt allele (minor allele) of GCKR (rs1260333) polymorphism was significantly lower in normal weight metabolically healthy participants than metabolically unhealthy normal weight (MUHNW) and obese children with and without cardiometabolic risk factor (P = 0.01). Frequency of ga allele of GCKR (rs780094) polymorphism was significantly higher in normal weight children with cardiometabolic risk factor than in their obese counterparts with cardiometabolic risk factor (P = 0.04). Frequency of cg and gg alleles (minor type) of MLXIPL (rs3812316) polymorphism in normal weight metabolically healthy participants was significantly higher than MUHNW (P = 0.04) and metabolically healthy obese children (P = 0.04). CONCLUSION: The findings of our study indicated that the minor allele of GCKR (rs1260333) single nucleotide polymorphisms (SNPs) could have pathogenic effect for obesity and cardiometabolic risk factors. Ga allele of GCKR (rs780094) SNPs had a protective effect on obesity. Minor alleles of MLXIPL (rs3812316) could have a protective effect for obesity and cardiometabolic risk factors.

The Effects of Crocin on 6-OHDA-Induced Oxidative/Nitrosative Damage and Motor Behaviour in Hemiparkinsonian Rats.

BACKGROUND: Crocin is considered to prevent oxidative stress-related diseases, such as ischemia and Alzheimer's. The aim of the present investigation was to evaluate the effects of crocin on motor behaviour and 6-OHDA-induced oxidative/nitrosative damage to the striatum in an experimental model of Parkinson's disease. METHODS: Left medial forebrain bundle was lesioned by microinjection of 6-OHDA (16µg in 0.2% ascorbate-saline). Crocin (30 and 60 mg/kg) was injected intraperitoneally three days before surgery until six weeks. Rotational behaviour and biochemical analysis were used to evaluate the effect of crocin in a unilateral 6-OHDA-induced model of Parkinson's disease. RESULTS: The contralateral rotations induced by apomorphine in 6-OHDA lesioned group were highly significant (P < 0.001) as compared to the sham group. Moreover, chronic administration of crocin at doses of 30 and 60 mg/kg over six weeks did not change the rotations. The TBARS and nitrite levels in the striatum were also significantly (P < 0.05) increased in lesioned group. Treatment with crocin at a dose of 60 mg/kg significantly decreased the nitrite levels (P < 0.05) in the striatum. CONCLUSION: Crocin at a dose of 60 mg/kg could be effective in preventing the nitrosative damage in the striatum. Further investigations using higher doses of crocin is suggested to get the full neuroprotective effects of crocin in Parkinson's disease.

The effect of regular exercise on antioxidant enzyme activities and lipid peroxidation levels in both hippocampi after occluding one carotid in rat.

Regular exercise has beneficial effects on cerebrovascular diseases; however, its biochemical mechanisms are not fully known. The purpose of this study was to determine antioxidant enzyme activities and lipid peroxidation of both hippocampi after applying exercise followed by occluding one common carotid. Wistar rats were divided into four groups of control, exercise, hypoperfusion and exercise-hypoperfusion (exe-hypo). In the exercise and exe-hypo groups, the rats were forced to run on a treadmill for 1 h a day for 2 months. The right common carotid of the animals in the (exe-hypo) group was occluded after the cessation of exercise. Surgery without occlusion of the carotid was applied on the control (without exercise) and exercise groups. All animals were sacrificed 1 and 24 h after surgery. The levels of malondialdehyde (MDA) and antioxidant enzyme activities in the hippocampi were measured. A significant interaction was observed between the exercise and hypoperfusion in both hippocampi (p<0.05). In comparison with the control group, there was significant elevation of catalase activity in the right and left hippocampus of the hypo group at 24 h (p<0.0001). Regarding the differences between the hemispheres, there was a significant increase in MDA and decrease in catalase activity in the left hippocampus in hypoperfusion group, but the exercise in the exe-hypo group succeeded in abolishing these alterations which were caused by hypoperfusion, This study shows that exercise pre-conditioning prevents some alterations in brain oxidant-antioxidant status which are induced by cerebral hypoperfusion. Further studies are needed in order to clarify the mechanism of exercise.

Comparison of SYBR Green and TaqMan methods in quantitative real-time polymerase chain reaction analysis of four adenosine receptor subtypes.

BACKGROUND: Real-time polymerase chain reaction (PCR) is based on the revolutionary method of PCR. This technique is the result of PCR enormous sensitivity and real-time monitoring combination. In quantitative gene expression analysis, two methods have more popularity, SYBR Green and TaqMan, SYBR Green is relatively cost benefit and easy to use and technically based on binding the fluorescent dye to double-stranded deoxyribonucleic acid (dsDNA) where TaqMan method has more expensive and based on dual labeled oligonucleotide and exonuclease activity of Taq polymerase enzyme. Specificity is the most important concern with the usage of any non-specific dsDNA-binding Dyes such as SYBR Green whiles more specificity showed by labeled oligonucleotide method such as TaqMan. In this study, we compared two common RT PCR methods, TaqMan and SYBR Green in measurement gene expression profile of adenosine receptors. MATERIALS AND METHODS: Gene expression profiles of A1, A2A, A2B and A3 Adenosine receptors were analyzed by optimized TaqMan and SYBR Green quantitative RT PCR in breast cancer tissues. Primary expression data was normalizing by B. actin reference gene. RESULTS: Efficiencies were calculated more than 95% for TaqMan and SYBR Green methods in all genes. The correlations between means of normalized data of each gene in two methods were positive and significant (P < 0.05). CONCLUSION: Data analysis showed that with the use of high performance primer and by use proper protocols and material we can make precise data by SYBR Green as TaqMan method. In other word by optimization of SYBR Green method, its performance and quality could be comparable to TaqMan method.

National report on the association of serum vitamin D with cardiometabolic risk factors in the pediatric population of the Middle East and North Africa (MENA): the CASPIAN-III Study.

OBJECTIVES: As the first, to our knowledge, nationwide study in the Middle East and North Africa (MENA), this study aimed to investigate the association of serum 25 hydroxy vitamin D [25(OH)D] levels with cardiometabolic risk factors in a nationally representative sample of the pediatric population in Iran. The second objective was to provide the prevalence of hypovitaminosis D and the percentiles of serum 25(OH)D in the study population. METHODS: This national population-based study was conducted among 1100 Iranian students living in 27 provinces in Iran. The association of 25(OH)D with each cardiometabolic risk factor was determined after adjustment for age, gender, body mass index, and waist circumference. RESULTS: Participants consisted of 1095 students (52% boys) with a mean age of 14.74 ± 2.61 y. The median 25(OH)D level corresponded to a vitamin D insufficiency level: 12.70 ng/mL in boys and 13.20 ng/mL in girls. Overall, 40% of participants were vitamin D deficient, and 39% had vitamin D insufficiency. There were no significant differences in these findings between boys and girls. Adjusted regression analysis revealed a significant weak inverse association of 25(OH)D with systolic blood pressure, diastolic blood pressure, total cholesterol, and low-density lipoprotein cholesterol. This association was significantly positive with high-density lipoprotein cholesterol, but not with fasting plasma glucose and metabolic syndrome. CONCLUSION: We found a considerably high prevalence of hypovitaminosis D in the pediatric population of a sunny region. Our findings also revealed an association of hypovitaminosis D with many cardiometabolic risk factors from childhood; these associations were independent of obesity indexes. It is of special concern that the highly prevalent disorders of low 25(OH)D and low high-density lipoprotein cholesterol in children and adolescents of the MENA region had significant association. The clinical importance of our findings needs to be confirmed in longitudinal studies.

Relationship between vitamin D receptor gene polymorphisms and migraine without aura in an Iranian population.

BACKGROUND: Inflammation has a key role in migraine pathophysiology. Vitamin D is an effective anti-inflammatory agent. The aim of this study was to investigate the association between migraine and two vitamin D receptor (VDR) polymorphisms (TaqI and FokI) and also the relationship between VDR polymorphisms and headache severity. METHODS: In this case-control study we assessed 103 patients with newly diagnosed migraine without aura and 100 healthy subjects. Patients filled headache impact test-6 (HIT-6) as a tool to assess headache severity. RESULTS: Genotype frequencies of VDR were significantly different between control and migraine patients. Heterozygote genotypes (Ff and Tt) were statistically more frequent in the migraine patients than the control subjects both for TaqI gene (P = 0.018; OR = 1.81, 95% CI = 1.03-3.18) and FokI gene polymorphisms (P = 0.001; OR = 2.91, 95% CI = 1.47-5.77). Also f and t alleles were more frequent in the migraine patients. Total HIT-6 score was significantly different between FokI heterozygote and homozygote patients (60.32 ± 1.87 versus 49.87 ± 2.69, resp., P = 0.004). CONCLUSIONS: In conclusion our results showed that TaqI and FokI gene polymorphisms are associated with migraine without aura in Iranians patients. Also headache severity in FokI heterozygote patients was significantly greater than in the homozygote patients.