The Effects of Conditioned Medium from Bone Marrow-derived Mesenchymal Stem Cells on EMT Markers

Abstract Introduction: Epithelial-Mesenchymal Transition in colorectal cancer cell is a critical process in which cells lose their epithelial properties and obtain mesenchymal characteristics, resulting in tumor cells and metastasis. This study attempted to work on the effects of conditioned medium from bone marrow-derived mesenchymal stem cells on EMT markers. Materials and Methods: In this study, HT29 was used  which is the colorectal cancer cell line. Cells were treated for 72 hours with BMSC-CM in order to induce EMT in HT29. The Real-Time PCR was used for evaluating EMT markers such as E-cadherin- β-catenin -vimentin-and transcription factors. Results: Inducing EMT in colorectal cancer cells caused morphological changes. It was manifested that E-cadherin is downregulated after induction of EMT with treated BMSC-CM. On the other hand, there were a significant increase in β-catenin, Vimentin, Snail and ZEB1 expression. Conclusion: Understanding the molecular basis of tumor metastasis is critical for colorectal cancer treatment. Findings demonstrated morphological alterations in consequence MSCs-CM activates induction of EMT. This process affects EMT markers of E-cadherina, Vimentin, β-Catenin and transcription factors of Snail and ZEB1. This model helps knowing cancer and metastasis pathway and also could be used in drug screening procedures

Large scale analysis of olfactory receptors with highly genetically variations in relation with specific anosmia

Olfaktorische Rezeptoren (ORs) sind heptahelicale G-Protein gekoppelte Rezeptoren und bilden im menschlichen Genom die größte Gruppe von Plasma Membran Rezeptoren. Der erste Schritt zur Wahrnehmung von Duftstoffen wird von ORs vermittelt. Dabei ist zu beachten, dass beim Menschen jeder Duftstoff unterschiedlich bewertet werden kann. ORs können eine große Breite von chemischen Strukturen wahrnehmen. Um mehr über den Zusammenhang zwischen ORs und deren chemischen Liganden zu verstehen, haben wir in dieser Studie Liganden-Rezeptor Interaktionen für humane ORs untersucht, von denen Kopienzahlvariation (CNVs) oder Segregierende Pseudogene (SPGs) bekannt sind. Dabei werden Liganden benutzt, von denen bekannt ist, dass sie von einigen Menschen nicht wahrgenommen werden können. Es wurden 18 ORs deorphanisiert, die SPGs und CNVs aufwiesen. Dabei zeigte sich, dass es keinen einfachen oder direkten Zusammenhang zwischen den molekularen Strukturen und aktiven Rezeptoren gibt

Pyrosequencing analysis for rapid and accurate detection of clarithromycin resistance-associated mutations in Iranian Helicobacter pylori isolates

Abstract Background Treatment of Helicobacter pylori (H. pylori) infection has become challenging following the development of primary antibiotic resistance. A primary therapeutic regimen for H. pylori eradication includes clarithromycin; however, the presence of point mutations within the 23S rRNA sequence of H. pylori contributes to clarithromycin resistance and eradication failure. Thus, we aimed to develop a rapid and precise method to determine clarithromycin resistance-related point mutations using the pyrosequencing method. Methods and results H. pylori was isolated from 82 gastric biopsy samples and minimal inhibitory concentration (MIC) was evaluated using the agar dilution method. Clarithromycin resistance-associated point mutations were detected by Sanger sequencing, from which 11 isolates were chosen for pyrosequencing. Our results demonstrated a 43.9% (36/82) prevalence in resistance to clarithromycin. The A2143G mutation was detected in 8.3% (4/48) of H. pylori isolates followed by A2142G (6.2%), C2195T (4.1%), T2182C (4.1%), and C2288T (2%). Although the C2195T mutation was only detected by Sanger sequencing, the overall results from pyrosequencing and Sanger sequencing platforms were comparable. Conclusions Pyrosequencing could be used as a rapid and practical platform in clinical laboratories to determine the susceptibility profile of H. pylori isolates. This might pave the way for efficient H. pylori eradication upon detection

Prevalence of Helicobacter pylori Infection in Dyspeptic Patients in Iran

Although Helicobacter pylori (H. pylori) infection has been known to be associated with several upper gastrointestinal disorders such as peptic ulcer and gastric cancer, the relationship between H. pylori infection and dyspeptic symptoms remains controversial. Furthermore, it is still not clear which factors are associated with H. pylori infection in the Iranian population. We investigated the prevalence of H. pylori infection in dyspeptic patients and factors associated with H. pylori infection in the Iranian population. In this cross-sectional study, 303 patients with dyspeptic symptoms underwent endoscopy. Clinical data and a questionnaire about gastrointestinal symptoms were collected from each patient. H. pylori status was evaluated by histological examination. Among the 303 patients, 263 (86.8%) were found to be positive for H. pylori. The prevalence of H. pylori infection decreased significantly with age. There was no difference in the prevalence of H. pylori infection between the patients with and those without a family history of gastroduodenal diseases. Among 250 patients with abdominal pain, 219 (87.6%) were infected with H. pylori. Among 211 patients with epigastric abdominal pain, 185 (87.7%) were infected with H. pylori. It was observed that belching was significantly associated with H. pylori infection (P = 0.03). Dyspepsia triggered by the consumption of tea was higher in H. pylori-positive patients than in H. pylori-negative patients (P = 0.03). The prevalence of H. pylori infection in dyspeptic patients was quite high in Iran. Belching and dyspepsia triggered by tea consumption was related with H. pylori infection

Antibiotic Susceptibility Profile of Helicobacter pylori Isolated from the Dyspepsia Patients in Tehran, Iran

Background/Aim: Helicobacter pylori is an important pathogen for gastroduodenal diseases. Infection with H. pylori can be limited by regimens of multiple antimicrobial agents. However, antibiotic resistance is a leading cause of treatment failure. The aim of this study has been to determine the resistance patterns of H. pylori strains isolated from gastric biopsies of patients with dyspepsia by agar dilution method, in Tehran, Iran. Patients and Methods : H. pylori isolates from patients with gastrointestinal diseases were evaluated for susceptibility testing by agar dilution method. Susceptibility testing was performed to commonly used antibiotics including clarithromycin, tetracycline, amoxicillin, metronidazole and ciprofloxacin. Results: Among 92 patients with dyspepsia, H. pylori strains were isolated from 42 patients. Seventeen (40.5%) of the isolates were resistant to metronidazole (MICs ≥ 8 μg/l), whereas one isolate (2.4%) was resistant to amoxicillin (MICs ≤ 0. 5 μg/ml) and ciprofloxacin (MICs ≤ 1μg/ml). The resistance rates to other antibiotics in H. pylori isolates are recorded as follows: clarithromycin 6 (14.3 %), tetracycline 2 (4.8%). In 5 of 42 resistant cases, combined resistance was found. Conclusions: These data suggest that metronidazole should be used among Iranian patients in first-line therapy with caution, and ciprofloxacin in association with amoxicillin and a proton pump inhibitor is more recommended

Synergistic therapeutic effect of mesenchymal stem cells and tolerogenic dendritic cells in an acute colitis mouse model

Cell-based therapy with tolerizing cells has been applied for the treatment of inflammatory bowel disease (IBD) in previous experimental and clinical studies with promising results. In the current study, we utilized the dextran sulfate sodium (DSS)-induced colitis model, to investigate if tolerogenic dendritic cell-mesenchymal stem cell (tDC-MSC) combination therapy can augment the therapeutic effects of single transplantation of each cell type. The effect of MSC and tDC co-transplantation on the severity of colitis was assessed by daily monitoring of body weight, stool consistency, and rectal bleeding, and compared with control groups. Moreover, the colon length, colon weight, myeloperoxidase (MPO) activity were measured and evaluated with histological analysis of colon tissues. The Treg cell percentage and cytokine levels in spleens and mesenteric lymph nodes (MLNs) were measured by flow cytometry and ELISA, respectively. The results showed co-transplantation of MSCs and tDCs was more effective in alleviating the clinical and histological manifestations of colitis than monotherapy, especially when compared with MSC alone. The protective effects of tDC-MSC were accompanied by the induction of Treg cells and increased the production of anti-inflammatory cytokines in spleens and mesenteric lymph nodes. Together, co-transplantation of MSCs and tDCs could be a promising and effective therapeutic approach in the treatment of IBD

The role of Helicobacter pylori and CagA in response to treatment in Iranian gastroesophageal reflux disease patients

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Increased Expression of <i>VANGL1</i> is Predictive of Lymph Node Metastasis in Colorectal Cancer: Results from a 20-Gene Expression Signature

This study aimed at building a prognostic signature based on a candidate gene panel whose expression may be associated with lymph node metastasis (LNM), thus potentially able to predict colorectal cancer (CRC) progression and patient survival. The mRNA expression levels of 20 candidate genes were evaluated by RT-qPCR in cancer and normal mucosa formalin-fixed paraffin-embedded (FFPE) tissues of CRC patients. Receiver operating characteristic curves were used to evaluate the prognosis performance of our model by calculating the area under the curve (AUC) values corresponding to stage and metastasis. A total of 100 FFPE primary tumor tissues from stage I–IV CRC patients were collected and analyzed. Among the 20 candidate genes we studied, only the expression levels of VANGL1 significantly varied between patients with and without LNMs (p = 0.02). Additionally, the AUC value of the 20-gene panel was found to have the highest predictive performance (i.e., AUC = 79.84%) for LNMs compared with that of two subpanels including 5 and 10 genes. According to our results, VANGL1 gene expression levels are able to estimate LNMs in different stages of CRC. After a proper validation in a wider case series, the evaluation of VANGL1 gene expression and that of the 20-gene panel signature could help in the future in the prediction of CRC progression