Screening of Myo7A mutations in Iranian patients with autosomal recessive hearing loss from west of Iran

Hearing loss (HL) is the most frequent neurosensory impairment. HL is highly heterogeneous defect. This disorder affects 1 out of 500 newborns. This study aimed to determine the role of DFNB2 locus and frequency of MYO7A gene mutations in a population from west of Iran. Methods: Thirty families investigated in Shahrekord University of Medical Sciences in 2014, genetic linkage analysis via four short tandem repeat markers linked to MYO7A was performed for two consanguineous families originating from Hamedan (family-13) and Chaharmahal-Bakhtiari (family-32) provinces of Iran, co-segregating autosomal recessive HL and showed no mutation in GJB2 gene in our preliminary investigation. All 49 coding exons and exon- intron boundaries of the MYO7A gene were amplified by PCR and analyzed using direct DNA sequencing. Results: Two of families displayed linkage to DFNB2. Family-13 segregated a homozygous missense mutation (c.6487G>A) in exon 48 that results in a p.G2163S amino acid substitution in C-terminal domain of the myosin VIIA protein. While family-32 segregated a homozygous nonsense mutation (c.448 C>T) in exon five, resulting in a premature truncation at amino acid position 150 (p.Arg150X) in the motor domain of this protein. Conclusion: Mutation frequency of MYO7A gene in different populations of Iran as well as cause of HL in most cases are still unknown and more extensive studies have to be done. © 2017, Iranian Journal of Public Health. All rights reserved

Aloe vera toxic effects: expression of inducible nitric oxide synthase (iNOS) in testis of Wistar rat

Objective(s): Nitric oxide (NO), a product of inducible nitric oxide synthase (iNOS), contributes in germ cell apoptosis. This study was aimed to evaluate the effects of Aloe vera gel (AVG) on male Wistar rat reproductive organ, serum NO level, and expression of iNOS gene in leydig cells. Materials and Methods: Adult male Wistar rats (n=36) were used for experiments in three groups. The experimental groups were orally administered with the AVG extract solution once-daily as follow: 150 mg.kg(-1); group A, 300 mg.kg(-1); group B, and only normal saline; group C (control group). They were mated with untreated females and the reproductive and chemical parameters were assessed for each group, including semen quality, serum testosterone, sperm fertility, gonad and body weight, serum NO concentration (by the Griess method), and iNOS gene expression (using RT-PCR). Results: The testes weight, serum testosterone, as well as sperm count and fertility of the AVG treated groups were significantly reduced when compared to the control (P<0.001). Concentration of serum NO was significantly increased (37.1 +/- 4.63 mu M) in the administrated group with higher AVG concentration, compared to the control group (P<0.001; 10.19 +/- 0.87 mu M); however, iNOS mRNA expression was increased in the treated animals (P<0.001). Conclusion: iNOS may play a functional role in spermatogenesis via apoptosis, reducing sperm count, but further studies are needed to illustrate the mechanisms by which AVG exerts its negative effects on spermatogenesis and sperm quality

Aberrant Expression of Three New Members of IL-1 Family (IL-36α, IL-37 and IL-38) in Patients with Multiple Sclerosis

Background & Objective: Multiple sclerosis (MS) is a neurodegenerative disease that is characterized by demyelination and neuronal dysfunction. The study of the expression of cytokine genes seems to be an appropriate option for assessing their potential for the onset and development of the disease. Therefore, in the present study, changes in the expression of three interleukins (IL-36α, IL-37, and IL-38) in MS and their association with clinical factors were investigated. Material & Methods: In this study, blood samples of 45 MS patients and 45 healthy controls were enrolled. Relative expression of genes was evaluated using the real-time PCR. Finally, the pattern of expression was analyzed by using statistical analyses. In the next step, the relationship between clinical characteristics and IL-36α, IL-37, and IL-38 expression was investigated. Results: The results showed that IL-36α and IL-37 increased in MS patient (1.8 and 3.5 fold respectively, P<0.01).  Moreover, a significant reduction in IL-38 was observed in MS samples (0.21 fold, P<0.01). The dysregulation in ILs expression was associated with clinical features. Conclusion: As a result, the two studied IL-1 family members (IL-36α and IL-37) may contribute to the progression of MS, so that increase in their expression is associated with EDSS disability and duration of disease. Another member of IL-1 family, IL-38 has a protective effect on MS diseases, and its downregulation is related to severe disability. Further investigation can help to determine accurate functional role of 3 interleukins in MS development

Association Between Helicobacter pylori cagA, babA2 Virulence Factors and Gastric Mucosal Interleukin-33 mRNA Expression and Clinical Outcomes in Dyspeptic Patients

Helicobacter pylori (H. pylori) infection has been reported in more than half of the world human population. It is associated with gastric inflammation and noticeable infiltration of the immune cells to the stomach mucosa by several cytokines secretion. IL-1 beta, IL-18 have been shown to contribute to H. pylori induced gastritis, but the details of inflammation and association of virulence factors remain unclear. IL-1 cytokine family has a new additional cytokine, Interleukin-33 (IL-33), which is contemplated to have an important role for host defense against microorganisms. H. pylori virulence factors important in gastritis risk are the cag pathogenicity island (cag-PAI) and babA. This study evaluated IL-33 mucosal mRNA expression levels in infected and uninfected patients and its relationship with bacterial virulence factors cagA, babA(2) and type of gastritis. Total RNA was extracted from gastric biopsies of 79 H. pylori-infected patients and 51 H. pylori-negative patients. Mucosal IL-33 mRNA expression levels in gastric biopsies were assessed using real-time PCR. Existence of virulence factors were detected by PCR. IL-33 mRNA expression was significantly higher in biopsies of H. pylori-infected patients compared to H. pylori-uninfected patients (P< 0.0001). Also there was a direct relationship between virulence factor bab-A2 and enhancement in IL-33 mRNA expression. Furthermore, IL-33 mRNA expression level was significantly lower in chronic gastritis patients compared with patients with active gastritis (P< 0.001). IL-33 may play a crucial role in the inflammatory response and induction of the chronic gastritis and severity of inflammatory changes in the gastric mucosa

Association Between Helicobacter pylori cagA, babA2 Virulence Factors and Gastric Mucosal Interleukin-33 mRNA Expression and Clinical Outcomes in Dyspeptic Patients.

Helicobacter pylori (H. pylori) infection has been reported in more than half of the world human population. It is associated with gastric inflammation and noticeable infiltration of the immune cells to the stomach mucosa by several cytokines secretion. IL-1β, IL-18 have been shown to contribute to H. pylori induced gastritis, but the details of inflammation and association of virulence factors remain unclear. IL-1 cytokine family has a new additional cytokine, Interleukin-33 (IL-33), which is contemplated to have an important role for host defense against microorganisms. H. pylori virulence factors important in gastritis risk are the cag pathogenicity island (cag-PAI) and babA. This study evaluated IL-33 mucosal mRNA expression levels in infected and uninfected patients and its relationship with bacterial virulence factors cagA, babA2 and type of gastritis. Total RNA was extracted from gastric biopsies of 79 H. pylori-infected patients and 51 H. pylori-negative patients. Mucosal IL-33 mRNA expression levels in gastric biopsies were assessed using real-time PCR. Existence of virulence factors were detected by PCR. IL-33 mRNA expression was significantly higher in biopsies of H. pylori-infected patients compared to H. pylori-uninfected patients (P<0.0001). Also there was a direct relationship between virulence factor bab-A2 and enhancement in IL-33 mRNA expression. Furthermore, IL-33 mRNA expression level was significantly lower in chronic gastritis patients compared with patients with active gastritis (P<0.001). IL-33 may play a crucial role in the inflammatory response and induction of the chronic gastritis and severity of inflammatory changes in the gastric mucosa

Screening LRTOMT gene (DFNB63 locus) in patients with recessive nonsyndromic hearing loss in hormozgan province, Iran

Background: Congenital hearing loss is the most common sensory disorder is modern societies. Two modes of syndromic and nonsyndromic genetic hearing loss can be seen. Autosomal recessive nonsyndromic inheritance hearing loss (ARNSHL) is the most common form of inheritance hearing loss. So, far mapping it, 95 loci of the 41 regions of deafness genes have been identified. Despite numerous studies in this field, for DFNB63 (Leucine rich transmembrane and O-methyltransferase or LRTOMT gene), few studies have been conducted. Thus, the locus mutations can help us to better understand the genes involved in hearing loss. Methods: 90 cases of hearing loss in Hormozgan province, Iran, were studied. DNA extracted via phenol-chloroform method. Polymerase chian reaction (PCR) was performed. Then, the product was used to perform polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and polymerase chain reaction-heteroduplex analysis (PCR-HA) methods. Samples with different bands were sequenced to determine the nucleotide changes. Findings: In different exons, 8 samples (each in a specific exon) were sequenced to determine the type of changes that shift single-strand conformation polymorphism bands. No change was found in nucleotide sequencing of exons in any of the groups. Conclusion: The results showed no relationship between non-syndromic deafness and LRTOMT gene mutations. As this gene is discovered in recent years, there are few studies on it. So far, only 5 mutation of this gene have been identified in the world that can determine pale relationship of the gene and hearing loss. © 2014, Isfahan University of Medical Sciences(IUMS). All rights reserved

A glance into the roles of microRNAs (exosomal and non-exosomal) in polycystic ovary syndrome

Polycystic ovarian syndrome (PCOS) is a common endocrine disorder in women of reproductive age. The clinical symptoms include hyperandrogenism, chronic anovulation, and multiple ovarian cysts. PCOS is strongly associated with obesity and insulin resistance. MicroRNAs (miRNAs) are a group of short non-coding RNAs that play a role in the post-transcriptional regulation of gene expression and translational inhibition. They play a vital role in the regulation of multiple metabolic and hormonal processes as well as in oocyte maturation and folliculogenesis in the female reproductive system. miRNAs can be used as diagnostic biomarkers or therapeutic targets because of their stability. The encapsulation of miRNAs in extracellular vesicles or exosomes contributes to their stability. Exosomes are constantly secreted by many cells and size of about 30 to 150 nm. Enveloping miRNAs exosomes can release them for cellular communication. The induced transfer of miRNAs by exosomes is a novel process of genetic exchange between cells. Many studies have shown that along with non-exosomal miRNAs, different types of exosomal miRNAs derived from the serum and follicular fluid can play an essential role in PCOS pathogenesis. These miRNAs are involved in follicular development and various functions in granulosa cells, apoptosis, cell proliferation, and follicular atresia. The present study aimed to comprehensively review the evidence on miRNAs and their affected pathways under both non-exosomal and exosomal circumstances, primarily focusing on the pathogenesis of PCOS

The Study of SLC26A4 Gene Causing Autosomal Recessive Hearing Loss by Linkage Analysis in a Cohort of Iranian Populations.

Sensorineural non-syndromic hearing loss is the most common disorder which affects 1 in 500 newborns. Hearing loss is an extremely heterogeneous defect with more than 100 loci identified to date. According to the studies, mutations in GJB2 are estimated to be involved in 50- 80% of autosomal recessive non-syndromic hearing loss cases, but contribution of other loci in this disorder is yet ambiguous. With regard to studies, DFNB4 locus (SLC26A4) can be classified as the second cause of hearing loss. So, this study aimed to determine the contribution of this locus in hearing loss as well as the frequency of SLC26A4 gene mutations in a population in the west of Iran. In this descriptive laboratory study, we included 30 families from the west of Iran with no mutation in GJB2 gene. Linkage analysis was performed by DFNB4 (SLC26A4) molecular markers (STR). The families with hearing loss linked to this locus were further analyzed for mutation detection. SLC26A4 gene exons were amplified and analyzed using direct DNA sequencing. In studied families, 2 families displayed linkage to DFNB4 locus. Identified mutations include mutation in exon 5 (c.416 G>T) and in splicing site of exon 7 (IVS-2 A>G or c.919-2 A>G)

Genetic linkage analysis of the DFNB63 locus in families with autosomal recessive nonsyndromic hearing loss from Hamadan and Kohgiluyeh and Boyer-Ahmad Provinces, Iran

Background: Hearing loss is a sensorineural impairment and is one of the most widespread congenital impairments with a prevalence of one in thousand among children. Studies have shown that 50 percent of congenital hearing loss have genetic causes and the remaining 50 percent are due to environment and unknown reasons; in addition, it is noted that this impairment is very heterogeneous. Almost 70 percent of cases are nonsyndromic with hearing loss presenting as the only impairment. About 80 percent of this type of hearing loss is inherited in recessive manner (ARNSHL). In this study, we determined the role of DFNB63 locus in a series of families in two western provinces of Iran. Methods: In this descriptive-laboratory study, to determine the prevalence of DFNB63 mutations in western provinces of Iran, we studied 150 individuals from 30 families in Hamadan and Kohgiluyeh and Boyer-Ahmad provinces. The selected families in this study were consanguineous, had at least 2 patients, and were negative for GJB2 mutations. Linkage analysis was performed using six appropriate short tandem repeats (STR) markers. Findings: With linkage analysis of selected families, no family was shown to be linked to the DFNB63 locus. It was shown that the LRTOMT mutations played no role in causing hearing loss in the studied families. Conclusion: The present study suggests that LRTOMT mutations may not be clinically important in causing autosomal recessive nonsyndromic hearing loss in the investigated provinces. © 2015, Isfahan University of Medical Sciences(IUMS). All rights reserved

Genetic linkage analysis of DFNB40 and DFNB48 loci in families with autosomal recessive non-syndromic hearing loss (ARNSHL) from western provinces of Iran

Background: Sensorineural hearing loss (SNHL) is the most common sensory disorder and 1 in every 500-1000 newborns is affected. Non-syndromic SNHL accounts for 70% of hereditary hearing loss and 80% of SNHL cases have an autosomal recessive mode of inheritance (ARNSHL). The Purpose of the recent study is genetic linkage analysis to determine the prevalence of DFNB40 and DFNB48 loci in studying families with ARNSHL from the western provinces of Iran. Methods: In this study, 60 families from 3 provinces of Iran involving Hamedan, Kohgiluyeh and Boyer-Ahmad and Chaharmahal and Bakhtiari with autosomal recessive non syndromic hearing loss were examined. The selected families in this study were consanguineous and had at least two patients. They also were negative for GJB2 mutations. Linkage analysis was performed by using 6 markers short tandem repeat (STR) for the DFNB40 locus and 7 markers STR for the DFNB48 locus. Findings: After examining different families, it was revealed that none of them showed linkage to the DFNB40 and DFNB48 loci. Conclusion: The recent study suggests that DFNB40 and DFNB48 loci might not play an important role in causing hearing loss in the mentioned provinces. However, further studies are necessary to determine more precisely the role of these loci in the Iranian population. © 2016, Isfahan University of Medical Sciences(IUMS). All rights reserved