A novel variant of SLC26A4 and first report of the c.716T>A variant in Iranian pedigrees with non-syndromic sensorineural hearing loss

The autosomal recessive non-syndromic hearing loss (ARNSHL) can be associated with variants in solute carrier family 26, member 4 (SLC26A4) gene and is the second most common cause of ARNSHL worldwide. Therefore, this study aims to determine the contribution of the SLC26A4 genotype in the hearing loss (HL) of 40 ARNSHL pedigrees in Iran. A cohort of the 40 Iranian pedigrees with ARNSHL, having no mutation in the GJB2 gene, was selected. The linkage analysis with five short tandem repeat (STR) markers linked to SLC26A4 was performed for the 40 ARNSHL pedigrees. Then, two out of the 40 pedigrees with ARNSHL that linked to DFNB4 locus were further screened to determine the variants in all exons of SLC26A4 gene by direct DNA sequencing. The 21 exons of SCL26A4 were analyzed for the two pedigrees. A known variant (c.716T>A homozygote), it is the first reported incidence in Iran, a novel variant (c.493A>C homozygote) were detected in the two pedigrees and pathogenesis of c.493A>C confirmed in this study with review 100 hearing ethnically matched controls by PCR-RFLP analysis. The present study suggests that the SLC26A4 gene plays a crucial role in the HL occurring in Iranian pedigrees. Also, the results probably support the specificity and unique spectrum of SLC26A4 variants among Iranian HL patients. Molecular study of SLC26A4 gene may lead to elucidation of the profile of the population-specific variants which has importance in diagnostics of HL

Virulence factors of Helicobacter pylori vacA increase markedly gastric mucosal TGF-beta 1 mRNA expression in gastritis patients

Objective: Helicobacter pylori (H. pylori) infection is the main cause of gastric inflammation. Regulatory T cells (Treg cells) suppress the activation and proliferation of antigen-specific T cells and mediate immunologic tolerance. TGF-beta 1 was shown to be secreted in a subset of Treg cells known as `Th3 cells'. These cells have not been sufficiently studied in context to H. pylon-induced inflammation in human gastric mucosa. In this study we therefore, aimed to investigate the expression of TGF-beta 1 in the context of H. pylori colonization in chronic gastritis, to examine the relationship between it and histopathologic findings and to compare it with virulence factors. Patients and methods: Total RNA was extracted from gastric biopsies of 48 H. pylori-infected patients and 38 H. pylori-negative patients with gastritis. Mucosal TGF-beta 1 mRNA expression in H. pylori-infected and uninfected gastric biopsies was determined by real-time PCR. Presence of vacA, cagA, iceA, babA2 and oipA virulence factors was evaluated using PCR. Results: TGF-beta 1 mRNA expression was significantly increased in biopsies of H. pylori-infected patients compared to H. pylori-uninfected patients. There was association between virulence factors and TGF-beta 1 mRNA expression. TGF-beta 1 mRNA expression in mucosa was significantly higher in patients with vacA s1 and s1m1. Conclusions: TGF-beta 1 may play an important role in the inflammatory response and promote the chronic and persistent inflammatory changes in the gastric. This may ultimately influence the outcome of H. pylon-associated diseases that arise within the context of gastritis and vacA may suffice to induce expression of TGF-beta 1 mRNA. (C) 2014 Elsevier Ltd. All rights reserved

Expression levels of mRNA cytokines of IL-17 and IL-23 in epithelialfiber of stomach inpatients with Helicobacter pylori using Real-Time PCR in Chahar Mahal and Bakhtiari province

زمینه و هدف: اینترلوکین های 17 و 23 در دفاع بر علیه برخی عفونت‌های مخاطی دستگاه گوارش نقش دارند و IL-17 باعث جذب نوتروفیل ها به محل عفونت شده و در ایجاد التهاب نقش دارد. مطالعه حاضر میزان بیان mRNA سیتوکاین های IL-17و IL-23در دو گروه بیماران گاستریتی با عفونت هلیکوباکترپیلوری و فاقد عفونت را به وسیله روش کمی Real-Time PCR بررسی می‌کند. روش بررسی: در این مطالعه مورد- شاهدی، از 58 بیمار دارای گاستریت با عفونت هلیکوباکترپیلوری و 50 بیمار مبتلا به گاستریت که فاقد عفونت بودند، توسط آندوسکوپی بیوپسی تهیه شد. بعد از استخراجmRNA و تبدیل آن به cDNA، میزان بیانmRNA مربوط به IL-17و IL-23در نمونه‌ها توسط Real-Time PCR اندازه گیری شد و بیان سایتوکاین ها در دو گروه آلوده و غیر آلوده با استفاده از تست Mann–Whitney مورد آنالیز قرار گرفت. یافته‌ها: ارتباط معنی‌داری بین میزان بیانIL-17 mRNA در افراد دارای گاستریت با عفونت هلیکوباکترپیلوری و افراد دارای گاستریت فاقد عفونت دیده نشد (941/0P=). همچنین ارتباط بین میزان بیان mRNA IL-23در بیماران دارای گاستریت با عفونت هلیکوباکترپیلوری و بیماران دارای گاستریت فاقد عفونت معنی دار نبود (076/0 P=). نتیجه‌گیری: نتایج این مطالعه نشان داد که میزان بیان mRNA سیتوکاین های IL-17و IL-23در بیماران دارای گاستریت با عفونت هلیکوباکترپیلوری در مقایسه با بیماران گاستریتی بدون عفونت بالاتر نمی‌باشد و در نتیجه ارتباط معنی داری بین دو گروه مورد مطالعه در این استان وجود ندارد؛ لذا می طلبد تا نقش دقیق سایتوکاین ‌های دیگر درگیر در بروز بیماری گاستریت جهت تعیین پیش آگهی و ارزیابی برنامه های درمانی بیشتر مشخص شود

Frequency of virulence factors in Helicobacter pylori-infected patients with gastritis

The outcome of Helicobacter pylori infection has been related to specific virulence-associated bacterial genotypes. The vacuolating cytotoxin (vacA), cagA gene, oipA and babA2 gene are important virulence factor involving gastric diseases. The objective of this study was to assess the relationship between virulence factors of H. pylori and histopathological findings. Material and methods: Gastroduodenoscopy was performed in 436 dyspeptic patients. Antrum biopsy was obtained for detection of H. pylori, virulence factors and for histopathological assessment. The polymerase chain reaction was used to detect virulence factors of H. pylori using specific primers. Results: vacA genotypes in patients infected with H. pylori were associated with cagA, iceA1 and iceA2. In the patients with H. pylori infection there was a significant relationship between cagA positivity and neutrophil activity (P = 0.004) and chronic inflammation (P = 0.013) and with H. pylori density (P = 0.034). Neutrophil infiltration was found to be more severe in the s1 group than in the s2 group (P = 0.042). Also was a significant relationship between oipA positivity and neutrophil activity (P = 0.004) and with H. pylori density (P = 0.018). No significant relationships were observed between other vacA genotypes and histopathological parameters. Conclusion: H. pylori strains showing cagA, vacA s1 and oipA positivity are associated with more severe gastritis in some histological features but virulence factors of H. pylori do not appear to determine the overall pattern of gastritis. (C) 2015 Elsevier Ltd. All rights reserved

Comparing the expression levels of mRNA for MMP-7 in gastric mucosa of patients with H. pylori infection and uninfected patients

Background and purpose: The expression of growth factors, proteolytic enzymes, fibrogenic factors, and cytokines are altered in Helicobacter pylori (H. pylori) infected gastric mucosa. Matrix metalloproteinases (MMP) are a family of zinc-dependent homologous enzymes digesting most of the components of the extracellular matrix and basement membrane and are involved in remodeling and functioning of the biological processes. The purpose of this study was to compare gene expression of matrix metalloproteinase-7 (MMP-7) in patients with H. pylori-infected and uninfected individuals with gastrointestinal diseases. Materials and methods: This study was conducted in 50 H. pylori-negative patients and 50 H. pylori-positive patients being admitted to Shahrekord Hajar Hospital due to gastrointestinal diseases in 2014. The participants’ demographic information was collected and sampling was done. First DNA was extracted, and then PCR was performed to check for the presence of 16sRNA and UreC. The RNA from each sample was also extracted and cDNA was prepared. Afterwards, the expression of MMP-7 was measured by real time-PCR using specific primers and probes. Results: MMP-7 mRNA expression was significantly higher in biopsies of H. pylori-infected patients compared to that in H. pylori-uninfected patients (P<0.0001). Conclusion: Increased expression of MMP-7 can be effective in inflammatory response and development of the disease. It could be used as a key marker for early diagnosis of gastrointestinal diseases and gastric cancer. © 2016, Mazandaran University of Medical Sciences. Engineering. All rights reserved

Effect of Allium sativum L. extract on acid and pepsin secretion in basal condition and stimulated with vag stimulate in rat

Background: Garlic is used world wide to give flavor to foods as pecols ect. by different nations specially by Iranian society. The dominant believe is, the garlic therapeutic effects on patients with gastric discomfort and digesting problems. Objective: in this study we aimed to investigate the garlic effect on gastric acid and pepsin in both basic and stimulated condition by electrical stimulus of voguesnerve in rat. Methods: in an experimental study two groups of 12 wistar rats were selected. to make the animal unconscious, 50mg/kg Sodium Tiopantal were given ip. and each rat was tracheostomized, laparatomized, gastrodeodonostomized and then 100 mg/kg of garlic extract dissolved in 9 saline was introduced to stomach of animal. The vagus nerves of both side of the neck released from carotid sheet and stimulated with 15millivolts, frequency 4 Herts and 1 milisecond wide. The stomach secretions then washed out in following stages; first base, second base, vagus stimulated conditions and reverse to base. The acid amounts of washing were measured by titrometry and pepsin amounts were measured with Anson technique. Results: the acid and pepsin secretion were significantly increased in rats received garlic extracts (p&lt;0.000 p&lt;0.05 respectively). Stimulation by vagus nerves in control group significantly increased the level of acid (p&lt;0.05) and pepsin (p&lt;0.05) and garlic treated group had similar secretion rate to stimulated group. Furthermore there were no significant relationship between sex of studied rats. Conclusion: consumption of garlic in the diet has lots of benefits for digestion of foods, so it can be beneficial for those patients with digestive problems, however it should be noted that people with digestive dysfunction related to increased level of gastric acid or pepsin must avoid garlic consumption. Therefore patient with gastritis and duodenal ulcer are not allowed to take garlic in their diet

The effect of Amirkabiria odoratissima M extract on gastric acid and pepsin secretion level in rat

چکیده: زمینه و هدف: گیاه کرفس کوهی از دسته گیاهانی است که به طور وسیعی در استانهای جنوب غرب ایران استفاده می شود. شمار زیادی از مردم بر این باورند که این گیاه برای ناراحتی های گوارشی مفید است. این مطالعه با هدف بررسی این گیاه استفاده می شود رو به افزایش است لذا این مطالعه با هدف بررسی اثر عصاره این گیاه بر میزان ترشح اسید و پپسین معده در موش صحرایی انجام شد. روش بررسی: این مطالعه بصورت تجربی بر روی سه گروه 8 تایی موش صحرایی صورت گرفت ]گروه کنترل، کرفس کوهی با دوز mg/kg 2/16 (گروه اول) و mg/kg 100 (گروه دوم). حیوانات پس از بیهوشی توسط تزریق داخل صفاقی mg/kg 50 تیوپنتال سدیم، تراکئوستومی، لاپاراتومی و گاسترودئودنوستومی شدند. عصاره گیاه کرفس کوهی با دوزهای mg/kg 2/16 و mg/kg 100 از طریق مجرای گاسترودئودنوستوم به درون معده حیوانات گروه های کرفس وارد شد. ترشحات معده به روش Wash Out شامل پایه اول و پایه دوم بدست آمد و اسید آن به روش تیتریمتری و پپسین به روش آنسون اندازه گیری شد. داده ها با استفاده از آزمونهای آماری t مستقل، t وابسته وANOVA مورد تجزیه و تحلیل آماری قرار گرفت. یافته ها: عصاره متانولی گیاه کرفس کوهی سبب کاهش معنی داری در میزان ترشح اسید معده در پایه اول و دوم در موشهای هر دو گروه کرفس کوهی نسبت به گروه کنترل گردید (01/0

Association between virulence factors of helicobacter pylori and gastric mucosal interleukin-18 mRNA expression in dyspeptic patients

Background: Helicobacter pylori (Hp) infection is associated with gastritis and marked infiltration of the gastric mucosa by several cytokines secreting inflammatory cells that contribute to sustain and expand the local inflammation. Different clinical expressions of the infection may reflect distinctive patterns of cytokine expression. IL-1 beta, TNF-alpha, IL-17 and IL-23 have been reported to be involved in Hp-induced gastric mucosal inflammation, but the details and association to different patterns of inflammation and virulence factors remain unclear. Methods: Total RNA was extracted from gastric biopsies of 51 Hp-infected patients and 44 Hp-negative patients. Mucosal IL-18 mRNA expression in gastric biopsies was determined by Real-Time PCR. Presence of virulence factors was evaluated using PCR. Results: IL-18 mRNA expression was significantly increased in biopsies of Hp-infected patients compared to Hp-uninfected individuals. There was no association between virulence factors and IL-18 mRNA expression. Also severity of mononuclear infiltration was significantly higher in gastritis patients with vacA (m1)-positive compare patients with vacA (m2)-positive. Conclusions: IL-18 may play an important role in the inflammatory response and promote the chronic and persistent inflammatory changes in the stomach. This may ultimately influence the outcome of Hp-associated diseases that arise within the context of gastritis. (C) 2013 Elsevier Ltd. All rights reserved

Altered Th17 Cytokine Expression in Helicobacter pylori Patients with TLR4 (D299G) Polymorphism

Helicobacter pylori (H. pylori) is associated with gastric ulcer and gastric adenocarcinoma. Polymorphisms in the host genes coding for Toll-like receptors (TLRs) may influence the innate and adaptive immune response to the infection, affecting the susceptibility to H. pylori or the disease outcomes. However, the details and association with different polymorphism and clinical expression of infection remain unclear. A case-control study consisting of 58 patients with H. pylori infection and 44 H. pylori uninfection was conducted. Genomic DNA was extracted and genotypes of TLR4 Asp299Gly polymorphism were assessed through polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Mucosal cytokines expression in H. pylori-infected and uninfected gastric biopsies was determined by real-time PCR. The expression of IL-6, IL-17, IL-21, IL-23 and TGF-β1 was signicantly higher in patients with D299G polymorphism in TLR4. But the expression of IL-18 between patients with single-nucleotide polymorphisms (SNPs) in TLR4 and patients with the wild-type allele was not signicant. In H. pylori-infected patients with gastritis, SNPs in TLR4 may alter cytokine expression toward Th17 immune response in the gastric mucosa and may have increased risk for the development of peptic ulcer

Virulence factors of Helicobacter pylori vacA increase markedly gastric mucosal TGF-β1 mRNA expression in gastritis patients

Objective: Helicobacter pylori (H. pylori) infection is the main cause of gastric inflammation. Regulatory T cells (Treg cells) suppress the activation and proliferation of antigen-specific T cells and mediate immunologic tolerance. TGF-b1 was shown to be secreted in a subset of Treg cells known as ‘Th3 cells’. These cells have not been sufficiently studied in context to H. pylori-induced inflammation in human gastric mucosa. In this study we therefore, aimed to investigate the expression of TGF-b1 in the context of H. pylori colonization in chronic gastritis, to examine the relationship between it and histopathologic findings and to compare it with virulence factors. Patients and methods: Total RNA was extracted from gastric biopsies of 48 H. pylori-infected patients and 38 H. pylori-negative patients with gastritis. Mucosal TGF-b1 mRNA expression in H. pylori-infected and uninfected gastric biopsies was determined by real-time PCR. Presence of vacA, cagA, iceA, babA2 and oipA virulence factors was evaluated using PCR. Results: TGF-b1 mRNA expression was significantly increased in biopsies of H. pylori-infected patients compared to H. pylori-uninfected patients. There was association between virulence factors and TGF-b1 mRNA expression. TGF-b1 mRNA expression in mucosa was significantly higher in patients with vacA s1 and s1m1. Conclusions: TGF-b1 may play an important role in the inflammatory response and promote the chronic and persistent inflammatory changes in the gastric. This may ultimately influence the outcome of H. pylori-associated diseases that arise within the context of gastritis and vacA may suffice to induce expression of TGF-b1 mRNA