Periplasmic expression and one-step purification of urease subunit B of Helicobacter pylori

UreB is one of the urease subunits of Helicobacter pylori and can be used as an excellent antigen candidate for H. pylori vaccination. Easy access to highly purified UreB protein, facilitate advances in therapeutic or preventive strategies. To achieve a simplified purification procedure, the present report represents a novel method of producing recombinant urease subunit B extracellularly. ureB gene from 26,695 standard strain was amplified by PCR and cloned into pET-26b(+) expression vector. UreB was expressed as a soluble, N-terminal pelB and C-terminal hexahistidine-tagged fusion protein (UreB-6His) and secreted into the periplasmic space of Escherichia coli. Expression of the recombinant UreB in E. coli BL21 (DE3) was induced by isopropylthio-β-d-galactoside (IPTG). Expression of UreB was confirmed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and western blot analysis using anti-His monoclonal antibody. UreB-6His protein was extracted from the periplasm by osmotic shock treatment and was purified in one step by Nickel affinity chromatography. In conclusion, the present protocol is easier to perform; more time effective and low cost than earlier methods. Keywords Helicobacter pylori –Urease subunit B–Cloning–Periplasmic expression Periplasmic expression and one-step purification of urease subunit B of Helicobacter pylori (PDF Download Available). Available from: https://www.researchgate.net/publication/227181089_Periplasmic_expression_and_one-step_purification_of_urease_subunit_B_of_Helicobacter_pylori [accessed Dec 09 2017]

Comparative effectiveness of aggressive intravenous fluid resuscitation with lactated Ringer’s solution and rectal indomethacin therapy in the prevention of pancreatitis after endoscopic retrograde cholangiopancreatography: a double blind randomised controlled clinical trial

Introduction : There is growing evidence indicating the aggressive intravenous fluid resuscitation (IVFR) can decrease the rate of pancreatitis; however, to the best of our knowledge it has not been well studied in a post-endoscopic retrograde cholangiopancreatography (post-ERCP) setting. Aim : To compare the effects of aggressive IVFR and rectal indomethacin (RI) in preventing pancreatitis after ERCP. Material and methods : This is a double blind randomised controlled clinical trial on 186 patients undergoing ERCP in Ahvaz, Iran. The inclusion criteria were ERCP for standard clinical indications such as choledocholithiasis, bile duct leak, and biliary obstruction. The IVFR group (n = 62) received a bolus of 20 ml/kg of body weight lactated Ringer’s solution (LRS) immediately after ERCP, followed by 3 ml/kg/h maintenance for 8 h. The RI group (n = 62) received 50 mg rectal indomethacin immediately before procedure and 12 h after ERCP. The control group (n = 62) did not receive any treatment. Results : Post-ERCP pancreatitis in IVFR, rectal indomethacin, and control groups occurred in 8 (12.9%), 16 (25.8%), and 20 (32.3%) patients (p = 0.036). Pancreatic pain was reported in 13 (21%), 21 (33.9%), and 27 (43.5%) patients in the IVFR, RI, and control group (p = 0.046). The serum amylase level increased over 24 h after intervention in all three groups. The mean serum amylase level 8 h after intervention in the IVFR patients was lower than the RI and control groups. Conclusions : Intravenous fluid resuscitation with LRS was more effective in preventing post-ERCP pancreatitis in comparison to the rectal indomethacin and control group

A Unique Profile of Adenomatous Polyposis Coli Gene Mutations in Iranian Patients Suffering Sporadic Colorectal Cancer

Objective: Colorectal cancer (CRC) is one of the most common and aggressive cancers worldwide. The majority of CRC cases are sporadic that caused by somatic mutations. The Adenomatous Polyposis Coli (APC; OMIM 611731) is a tumor suppressor gene of Wnt pathway and is frequently mutated in CRC cases. This study was designed to investigate the spectrum of APC gene mutations in Iranian patients with sporadic colorectal cancer. Materials and Methods: In this descriptive study, Tumor and normal tissue samples were obtained from thirty randomly selected and unrelated sporadic CRC patients. We examined the hotspot region of the APC gene in all patients. Our mutation detection method was direct DNA sequencing. Results: We found a total of 8 different APC mutations, including two nonsense mutations (c.4099C>T and c.4348C>T), two missense mutations (c.3236C>G and c.3527C>T) and four frame shift mutations (c.2804dupA, c.4317delT, c.4464_4471delATTACATT and c.4468_4469dupCA). The c.3236C>G and c.4468_4469dupCA are novel mutations. The overall frequency of APC mutation was 26.7% (8 of 30 patients). Conclusion: This mutation rate is lower in comparison with previous studies from other countries. The findings of present study demonstrate a different APC mutation spectrum in CRC patients of Iranian origin compared with other populations

An investigation into the sensitivity of endoscopic ultrasound in the diagnosis of malignant bile duct in patients with idiopathic acute pancreatitis

Introduction and Objective: Acute pancreatitis (AP) is an inflammatory process of the pancreas characterized by abdominal pain and increased pancreatic enzymes. This disease is diagnosed clinically. Endoscopic ultrasound (EUS), which is a technique with high sensitivity and specificity, is used to diagnose biliary disease. This study aimed to determine the sensitivity of EUS in the diagnosis of malignant bile duct in patients with idiopathic AP. Methods: This descriptive study was performed on 146 patients with pancreatitis hospitalized in the gastrointestinal tract section of the Imam Khomeini Hospital of Ahwaz Jundishapur University of Medical Sciences. The collected data were analyzed by the SPSS 22.0 and the significance level of the test was <0.05. Results: According to the results, 79 (54%) out of the 146 patients were female and 67 (46%) were male. The mean and standard deviation of the patients' age were 52.5 and 19.6 years, respectively. The findings showed that the sensitivity and specificity of the EUS were 33% and 99%, respectively. Compared to the endoscopic retrograde cholangiopancreatography (ERCP), the sensitivity and specificity of the abdominal ultrasound were 62% and 62.5%, respectively. Compared to the ERCP, the sensitivity and specificity of EUS were 92% and 50%, respectively. Conclusion: The findings of this study showed that the sensitivity and specificity of EUS were higher than those of abdominal ultrasound. Moreover, EUS was the preferred method to detect common bile duct stones (CBDS)

Relationship between serum ferritin level and transient elastography findings among patients with nonalcoholic fatty liver disease

Introduction: Nonalcoholic fatty liver disease (NAFLD) is raising prevalence among children, and adolescence population in developed and developing countries as a major public health concern. The present study aims to determine the relationship between serum ferritin level and transient elastography findings in patients suffering from NAFLD. Materials and Methods: The demographic and biochemical profile of included individuals such as body mass index, age, level of serum transaminases, fasting blood sugar, lipid profile, and serum ferritin level were determined and a transient elastography was performed for all of them. Results: The mean serum ferritin level among men with mild and advanced liver stiffness was 154 ± 97 and 244 ± 214, respectively (P < 0.001), which showed a meaningful relationship. These figures among female patients with mild and advanced liver stiffness included 79 ± 91 and 161 ± 103, respectively (P = 0.003) and again revealed a significant relationship. The cutoff values of ferritin with 90% accuracy for differentiation of mild from advanced liver stiffness among male and female patients were determined as 255 ng/ml and 135 ng/ml, respectively. These cutoff values for ruling out of advanced liver stiffness with 90% accuracy among both sexes were 72.5 ng/ml and 65.5 ng/ml, respectively. Conclusion: The finding of this study revealed a significant relationship between serum ferritin level and liver stiffness among NAFLD patients, and if these results repeated in further investigations, it could be advisable to measure serum ferritin level for predicting possibility of advanced liver fibrosis