Journal of Arak University of Medical Sciences

Analysis of Genetic Variation of rs1542705 Marker in SMPD1 Gene Region as an Informative Marker for Molecular Diagnosis of Niemann-Pick Disease in Isfahan Population

Sadegh Vallian Borujeni

Background: Niemann-Pick disease (NPD) refers to a group of lysosomal storage diseases that causes abnormal metabolism of lipids. One of the genes that play a role in the pathogenesis of this disease is SMPD1. To date, more than hundred disease- causing mutations have been identified in SMPD1 gene. Due to the large number of mutations in this gene, direct analysis of the mutations is costly and time-consuming. Therefore, indirect linkage analysis using polymorphic markers as an alternative method for molecular diagnosis of the mutations has been recommended. In the present study, allele frequency of rs1542705 genetic marker was analyzed in the Iranian population. The aim was to determine the polymorphic information content (PIC) and the possibility of its application in indirect diagnosing of NPD.

Materials and Methods: After bioinformatics analysis of the SMPD1 gene region, rs1542705 marker was selected for genotyping in Isfahan population. In order to calculate the allele and genotype frequency of the marker, molecular tests were done on 113 DNA samples of unrelated healthy individuals by using ARMS-PCR technique. Finally, the information related to the genotype of the individuals was statistically analyzed using Powermarker and Genepop software.

Results: The analyses showed that the studied population was in accordance with Hardy-Weinberg equilibrium. Allele frequency of rs1542705 marker for T and C alleles was 71.24% and 28.76%, respectively, and the heterozygosity of the marker was 43.36%. Also, polymorphic information content (PIC) was 0.325.

Conclusion: The results of this study showed that rs1542705 marker could be considered as an informative marker for molecular diagnosis of Niemann- Pick disease using linkage analysis in the studied population.

Statistical Analysis of Failure Time of the Molar Tooth Using Goodness-of-Fit Technique for Interval Censored Data

Reza Pakyari

Background: The failure time of permanent tooth is of the form of interval censored since the exact time of tooth decay is not available and it is only known that tooth decay occurs between two consecutive visits. There are a few techniques available in the literature for the problem of goodness-of-fit for interval censored data. In this paper, we propose a new goodness-of-fit testing procedure for interval censored data and employ this for the failure time of the first permanent molar tooth (sixth tooth) data.

Materials and Methods: Two methods of goodness-of-fit for interval censored data that are based on randomly generated data from each interval and averaging over the test statistics or over the p-values are employed for the failure time of the first permanent molar tooth data.

Results: The mean of the failure time of the first permanent molar tooth is found to be at 95 months. The p-values of the two goodness-of-fit testing procedures for the Weibull, log-normal and gamma models are calculated.

Conclusion: By comparing the p-values, the log-normal model is considered as the best model to describe the failure time of the first permanent molar tooth data.

Analysis of Y Chromosome Microdeletions and Mutation in Exon7 of the STAG3 Gene in Iranian Infertile Men with Idiopathic Non-Obstructive Azoospermia

Reza Mirfakhraei

Background: Azoospermia is defined as the absence of sperm in the semen and is divided in two types; obstructive and non-obstructive azoospermia. Non-obstructive azoospermia include approximately 60% of azoospermia patients. Several genetic and environmental factors can be involved in the development of non-obstructive azoospermia. Until now, several genes have been introduced as the causing factor of the azoospermia that are involved in spermatogenesis and testicular development. These genes are located on Y and/or autosome chromosomes .The aim of the present study was to investigate Y chromosome microdeletions and STAG3 gene mutations in Iranian males with non-obstructive azoospermia.

Materials and Methods: In this study, peripheral blood samples were obtained from 122 men with idiopathic non-obstructive azoospermia and 100 Normo-sperm men who had at least one child and DNA was extracted. Samples were investigated for the presence of Y chromosome microdeletions by Multiplex PCR. Then, existence of probable mutations in exon 7 of STAG3 gene was investigated using MSSCP (multi-temperature single-strand conformational polymorphism) method.

Results: 13 patients (10.66%) had Y chromosome microdeletions, but none of the subjects showed mutation in exon 7 of STAG3 gene. The Y chromosome microdeletions were found in none of the control individuals.

Conclusion: The results showed that Y chromosome microdeletions are the most important cause of non-obstructive azoospermia and should be considered as the main candidate for male infertility diagnostic tests. Mutations in the STAG3 gene are not common among non-obstructive azoospermia patients.

To Study the Effect of Calcium Plus VitaminD Supplementation on Nitric Oxide and Biomarkers of Oxidative Stress in Overweight and Obess Women with Polycystic Ovary Syndrome

Mehri jamilian

Background: Few studies have examined the effects of calcium plus vitamin D supplementation on metabolic profiles in overweight women with polycystic ovary syndrome (PCOS).This study was conducted to determine the effects of calcium plus vitamin D supplementation on metabolic profiles among overweight women with PCOS.

Materials and Methods: This randomized double-blind placebo-controlled clinical trial was conducted among 104 overweight and obess women with PCOS. Participants were randomly assigned into four groups to receive: 1) 1000 mg/d calcium+ vitamin D placebo; 2) 50000 IU/weeks vitamin D; 3) 1000 mg calcium/d and 4) calcium placebo + vitamin D placebo for 8 weeks. Fasting blood samples were taken at baseline and after 8 weeks intervention to measure metabolic profiles.

Results: After administrating calcium plus vitamin D supplementation, a significant decrease was observed in serum insulin (-3.3±11.4 vs.-1.1±8.5, -1.4±3.0 and +3.1±6.1 µIU/ml, p=0.03), insulin resistance (-0.7±2.4 vs.-0.3±2.5, -0.2±0.6 and +0.8±1.9, p=0.04), malondialdehyde (-0.6±0.3 vs.-0.1±0.2, -0.5±0.2 and +0.6±0.4 µmol/L, p=0.009) and a significant rise was seen in total antioxidant capacity (+35.2±21.7 vs.+22.5±37.7, +21.1±34.1 and -153.8±65.2 mmol/L, p=0.006) and glutathione (+216.0±79.3 vs.-47.5±60.2, +3.9±31.3 and -160.8±82.9 µmol/L, p=0.001) compared with vitamin D, calcium and placebo groups.

Conclusion: In conclusion, calcium plus vitamin D supplementation for eight weeks among women with PCOS had beneficial effects on metabolic profiles.

Irisin Response to Two Types of Exercise Training in Type 2 Diabetic Male Rats

Mousa Khalafi

Background: Irisin is an exercise-induced myokine that is reduced with type 2 diabetes and improves insulin resistance via the browning of white adipose tissues. However, irisin response to two types of exercise in patients with type 2 diabetes is unknown.

Materials and Methods: In this study, 22 diabetic Wistar rats (Induced by high-fat diet and injections Stz) were randomly assigned to 3 groups: high intensity interval exercise (HIIT), low intensity continuous training (LICT) and control (C). Both HIIT and LICT groups trained on the treadmill 5 sessions per week for 8 weeks. Blood samples were taken 24 hours after the last training session and plasma irisin, insulin and glucose levels were measured. ANOVA and Tukey post hoc tests were used to analyze data and the level of significance has been considered at p≤0.05.

Results: Data analysis showed that plasma irisin levels in the HIIT group were significantly increased compared to the control group (p<0.05). However, there was no significant difference between other groups (p>0.05). Plasma glucose in both HIIT and LICT groups was significantly decreased compared to the control group (p<0.05). But plasma insulin levels and insulin resistance index were not different between groups (p>0.05).

Conclusion: The results of this study showed that exercise training can increase plasma irisin in rats with type 2 diabetes. However, these changes are partially dependent on the type of exercise training.

Methodology for Designing Models Predicting Success of Infertility Treatment

Koorosh Kamali

Background: The prediction models for infertility treatment success have presented since 25 years ago. There are scientific principles for designing and applying the prediction models that is also used to predict the success rate of infertility treatment. The purpose of this study is to provide basic principles for designing the model to predic infertility treatment success.

Materials and Methods: In this paper, the principles for developing predictive models are explained and then the design of such models in infertility treatments is described in more details by explaining one sample.

Results: The important principles for models that firstly are described are: identifying and defining the purpose, expected function of model, input data that will be used to develop a model: type of intervention or diagnostic procedures that can lead to changes in the samples and output definition or expected result of model function. Further, characteristics of predictive factors in final model, drawing the information flowchart, internal and external validation and attention to the analysis programme of results are the important subjects that have been described.

Conclusion: If predictive models are used properly, can help treatment team and patients to achive best treatment in ART.

The Effect of Active VitaminD on Treatment of Proteinuria in Patients with Diabetic Nephropathy without Vitamin D Deficiency

Nader Saeedi

Background: The renin-angiotensin system(RAS) has a major role in development of diabetic nephropathy and blocking of RAS by inhibitors and blocking of angiotesin receptors is standard treatment for preventing kidney disease and proteinuria. It is reported that VIT-D analogues are able to suppress renin exertion and improve proteinuria. The aim of this study is to evaluate the effect of VitD analogue (calitriol) on reducing proteinuria in patients with diabetic nephropathy.

Materials and Methods: In this clinical trial study, 132 eligible patients that had diabetic nephropathy and hadn’t vit D deficiency were selected. The patients were divided into two equal groups. First group received the combination of losartan 25mg twice daily and calcitriol 0.25mg and second group received losartan 25 mg twice daly alone for 3 months. The FBS, lipid profile, ESR-CRP BUN, Cr, HbA1c, Ca, P, and 24 hours urine protein were evaluated in all patients at beginning and end of study and the results were statistcally compared.

Results: The 24-hour urine protein in losartan and calitriol group was improved compared to losartan. This difference was statistically significant (p=0.003). As well as, in kidney function (BUN, Cr) in the losartan and calcitriol group compared to losartan alone was significantly improved(p<0.05).

Conclusion: Combination of calcitriol with angiotesin receptor blockers(ARBs) is more effective than ARBs alone in improvement of proteinuria and real function.

Genetic Linkage Analysis of DFNB2 Locus with Autosomal Recessive Hearing Loss in Families Negative for GJB2 Mutations in Khuzestan Province

Seyed Reza Kazemi Nezhad

Background: Hearing loss is a common sensory impairment in humans which half of its causes are genetic reasons. Genetic hearing loss can be divided into the two types of syndromic and non-syndromic, which 80% of non-syndromic cases is Autosomal Recessive Non-Syndromic Hearing Loss. The aim of the present research is to determine the contribution of DFNB2 locus (MYO7A gene) in causing an autosomal recessive hearing loss in the one group of the deaf families of Khuzestan province.

Materials and Methods: This study was conducted on 26 families with autosomal recessive hearing loss (with 4 patients) and negative for GJB2 mutations in Khuzestan province. 22 families suffered from ARNSHL and 4 families suffered from Usher syndrome. Linkage analysis was performed by using STR (Short Tandem Repeat) markers related to DFNB2 locus. Each family’s genotype was determined by PCR-PAGE method. Furthermore, haplotypes drawing and LOD score calculations were performed.

Results: From 26 families with hearing loss participating in this research, following genetic linkage analysis and haplotypes drawing, two families (7.7% of the families) showed linkage to DFNB2 locus. One family (4.5%) suffered from ARNSHL and another family suffered from Usher syndrome.

Conclusion: The results of the present research show that the contribution of DFNB2 locus in causing hearing loss in the population of Khuzestan province was similar to other studies conducted in Iran and this locus with other important loci should be considered to check in the hearing loss panel.

The Effect of Folate Supplementation on Ghrelin of Stomach and Insulin Level of Serum in Male Wistar Rats during 10 Weeks of High Intensity Interval Training

Ali Gorzi

Background: High intensity training can lead to lower the appetite. So, the purpose of this study was to investigate the effect of folate supplementation on ghrelin level of stomach and insulin level of serum in male wistar rats during 10 weeks of high intensity interval training (HIIT).

Materials and Methods: Twenty seven male Wistar rats (weight= 203.94±27.34 gr, Age: 9 weeks) after one week familiarization, were randomly divided into four groups: control (n=6), folate supplementation (n=6), (HIIT) (n=7) and HIIT+ folate supplement (n=8). HIIT training protocol started with 30 m/min running on treadmill for 1 min with 10 reps and 2 min active rest at the first week and reached to 75-80 m/min for 1 min with 7 reps and 3 min active rest at last 3 weeks. Acylated ghrelin level of stomach tissue and serum level of insulin were assayed by ELISA kit.

Results: The results of Kruskal-vallis analysis showed that the ghrelin level of stomach was increased significantly (p=0.001) in folate+HIIT in compare with HIIT group. Also, insulin level of serum was decreased significantly (p=0.001) in folate +HIIT in compare with control and HIIT groups.

Conclusion: Based on our results, folate supplementation during high intensity interval training, increased the ghrelin of stomach and decreased the insulin level of serum. So, it seems that folate supplementation can prevent from losing appetite in athletes who train with high intensity training with interval type.

Prevalence of Speech Disorders in Arak Primary School Students, 2014-2015

Akram Valizadeh

Background: The speech disorders may produce irreparable damage to childs speech and language development in the psychosocial view. The voice, speech sound production and fluency disorders are speech disorders, that may result from delay or impairment in speech motor control mechanism, central neuron system disorders, improper language stimulation or voice abuse.

Materials and Methods: This study examined the prevalence of speech disorders in 1393 Arakian students at 1 to 6th grades of primary school. After collecting continuous speech samples, picture description, passage reading and phonetic test, we recorded the pathological signs of stuttering, articulation disorder and voice disorders in a special sheet.

Results: The prevalence of articulation, voice and stuttering disorders was 8%, 3.5% and%1 and the prevalence of speech disorders was 11.9%. The prevalence of speech disorders was decreasing with increasing of student’s grade. 12.2% of boy students and 11.7% of girl students of primary school in Arak had speech disorders.

Conclusion: The prevalence of speech disorders of primary school students in Arak is similar to the prevalence of speech disorders in Kermanshah, but the prevalence of speech disorders in this research is smaller than many similar researches in Iran. It seems that racial and cultural diversity has some effect on increasing the prevalence of speech disorders in Arak city.