Antigenic and immunogenic evaluation of Helicobacter pylori FlaA epitopes

Objective(s): Helicobacter pyloriare among most common human pathogens affecting at least half of the world’s population. Mobility is one of the important primary factors in bacterial colonization and invasion. The purpose of this research is cloning, expression, and purification of FlaA protein specific epitopes in order to evaluate their antigenicity and immunogenicity. Materials and Methods: The antigenic region of the flaA gene was bioinformatically predicted using Epitope mapping software’s and the predicted epitopes were expressed in a prokaryotic expression vector. The antigen was injected into the animal model (mice BALB/c) and some indicators including IgG1, IgG2a, IgA, IFN-γ, and IL 5 were measured. Results: The immunogenicity studies in animal models by measuring serum antibodies (IgG1, IgG2a, and IgA) and cytokines (IFN-γ and IL5) revealed that the rFlaA induces a proper immune response in animal models. Conclusion: The recombinant FlaA protein is antigenic and immunogenic. Therefore, it might be used in order to design of specific diagnostic kits and recombinant vaccines against H. pylori

Dysentery caused by macrolide and fluoroquinolone resistant Campylobacter coli in central area of Iran

Background: Campylobacter genus is considered some of the most important agents of bacterial gastroenteritis worldwide. Campylobacter coli (C. coli) is accounted to at least 25% of all Campylobacter related diarrheal diseases moreover, C. coli infections can result in severe complications, such as bacteremia, sepsis, meningitis and spontaneous abortion. Finally, there is evidence that the frequency of antimicrobial resistance is higher in C. coli, when compared to C. jejuni. There is no data regarding the frequency and antibiotic resistance profile of C. jejuni isolated from human gastroenteritis samples. The present study aimed to determine the frequency and antibiotic resistance patterns of Campylobacter coli isolated from infectious diarrhea samples. Methods: In a descriptive cross-sectional study, 200 infectious diarrhea samples collected in Arak University of Medical Sciences Hospitals, Markazi Province, Iran, from May to November 2015 were subjected to the study. In order to identify C. coli modified Gram stain, modified charcoal-cefoperazone-deoxycholate agar (mCCDA) and Brucella agar media with filter and CeuE gene polymerase chain reaction (PCR) were accomplished. Antibiotic resistance against tetracycline, erythromycin, ciprofloxacin, ampicillin and gentamicin was evaluated phenotypically and genotypically. Results: In total, out of 200 modified gram stained samples, 2 cases (1%) of C. coli were identified. Cultivating methods using mCCDA medium found 2 isolates (1%), 3 isolates (1.5%) were grown on Brucella agar with filter and 5 cases (2.5%) were determined as C. coli using PCR assay. Antibiotic resistance was observed in 5 cases against tetracycline, erythromycin and ciprofloxacin (100%), in 4 cases against ampicillin (80%), in 2 cases against gentamicin (40%), in 5 cases with CmeB, 23srRNA mutation in, qnrS, tet (o) (100%), in 4 cases with gyrA4 (80%), in 3 cases with gyrA5 (60%), in 5 cases with gyrA6 (100%), in 4 cases with Oxa61 (60%) and in 1 case with aphA-3-1 (20%). Conclusion: In this present study C. coli with low prevalence and entire resistance to ciprofloxacin and erythromycin which are the first line antibiotic for the treatment of campylobacter gastroenteritis is introduced as a causative agent of gastroenteritis in patients at central part of Iran

Detection of human CMV PP65 protein in glioma brain tumors with immunohistochemistry method

Background: Human cytomegalovirus (HCMV) may play a role in the development of glioma disease that is one of the most common brain tumors. Objective: The aim of this study was to detect human CMV in patients with glioma in Imam Khomeini hospital, Tehran. Methods: This experimental study was conducted on paraffin-embedded tumor samples of 18 patients referred to Imam Khomeini hospital in 2012. Immunohistochemistry (IHC) was performed with monoclonal antibody specific for HCMV PP65 protein and the samples were assessed using a light microscope. Findings: Of 18 patients, 13 (72.2%) were positive for HCMV PP65 protein and four of them expired. Conclusion: With regards to the results, more comprehensive studies are recommended for detection of HCMV in patients with glioma using different diagnostic methods. Keywords: Cytomegalovirus, Immunohistochemistry, Gliom

Bioinformatic Prediction of miRNAs Targeting NOTCH1 and HBx Genes in Chronic Hepatitis B-Induced Hepatocellular Carcinoma

Abstract Background: Hepatocellular carcinoma (HCC) is the third major cause of cancer death worldwide. Hepatitis B virus (HBV) and HBx gene play an important role in the development of HCC by influencing signaling pathways. Since there is no detectable symptom in the early phase of HCC, there is need to find new HCC-specific markers with high sensitivity for early detection and diagnosis of HCC. On the other hand, by the advent and development of bioinformatic sciences, it is now possible to predict miRNAs as biomarkers, and their targets. Therefore, in the present study, based on the results of the bioinformatic software applications with different algorithm, we selected the miRNA targeting HBx and NOTCH1 mRNAs according to higher score, suitable connection with target gene and confirming them in more softwares. Materials and Methods: First, the sequences of NOTCH1 and HBx genes were retrieved from NCBI. Afterwards, several software applications such as TargetScan, mirWalk, miRBase, Miranda, PicTar, miRVir, and DIANA were applied to predict miRNAs. Results: Based on the high scoring by bioinformatics softwares and suitable targeting, miR-34a were selected to target NOTCH1 and miR-6510, miR-5193 and miR-214 were chosen to targetHBX gene. Conclusion: Because of tumor suppression roles of miR-214 and miR-34a, they probably could be used as therapeutic strategy in cancer researches. It is also seems that the miR-5193 could act as a specific marker in Hepatocellular carcinoma

To Select the Appropriate Reference Gene for Normalizing the Quantitative Data to Assess MicroRNAs in Plasma Samples of Patients with Gastric Cancer

Abstract Background: Circulating microRNAs are promising biomarkers in diagnosis and assessment of cancerous patients. Quantitative Real-time PCR assay is a sensitive test for evaluating the levels of miRNAs expression. Nevertheless, there is no concurrence on selecting appropriate reference genes for qPCR analysis of miRNAs in circulation. Therefore, the current study aimed to select a suitable reference gene for normalizing the RT-qPCR assay results in plasma samples of patients with gastric cancer. Materials and Methods: Based on previously published studies, three molecules SNORD47, U6 RNA, and miR-103 were selected as the candidate reference genes. After RNA extraction from plasma samples of 40 patients with gastric cancer and 40 healthy individuals, expression levels of these molecules were evaluated using Real-time PCR method. Results: The results showed that the developed assays are able to diagnose their specified targets by a suitable linear range. By comparing patients and control groups, although the expression levels of miR-103 molecule were not equal between the two groups (p= 0.017), SNORD47 and U6 RNAs had similar expression levels. However, the variations of SNORD47 expression were lower that U6 RNA. Conclusion: Based on the results of the current study, the SNORD47 molecule has a stable expression levels in plasma samples of patients with gastric cancer and normal individuals and can be used as an appropriate reference gene for normalizing the quantitative data of qPCR assay

Frequency Determination of Ureaplasma and Mycoplasma Genitalium Species in Female with Vaginitis Infection using Real- Time PCR

Abstract Background: Ureaplasma and M. genitalium species belong to a kind of bacteria that are sexually transmitted and are the possible cause of pelvic inflammatory disease and nongonococcal urethritis, and et al. The aim of this study was to determine the urea plasma and Mycoplasma genitalium species frequency in women with vaginal infection and various sexual partners who referred to women, s health promotion and treatment center in Arak. Materials and Methods: Endocervical swab samples from 110 women with vaginal infections referred to women’s health promotion and treatment center in Arak, were prepared. Patients’ personal information and identities during reception process were registered. The samples were transferred to the laboratory in the transport environment and after DNA extraction, were evaluated according to Real-time PCR assay. Results: Urea plasma and Mycoplasma genitalium bacteria existed in 96(87.27%) and 4(3.63%) of patients, respectively. Among them, 4 cases had both bacteria infections. The amount of isolation in young women between 30-39 years old was more than others. Conclusion: The results show that the colonization of urea plasma species in adult women is 40-80% and in studied group is 87.27%. These results indicate that with due attention to the increasing number of sexual partners and the increase of sexual activity, the urea plasma colonization of women will increase. In view of the potential influence of mycoplasma species on side effects resulted from pregnancy infection of mothers and mortality, on-time diagnosis and treatment will be increasingly essential

SARS-CoV-2 Viral Shedding and Associated Factors among COVID-19 Inpatients and Outpatients

Background. According to the contagious ability of the new virus, SARS-CoV-2, characterization of viral shedding duration in the period of infection is highly valuable in terms of providing quarantine guidelines and isolation policies. Therefore, we aimed at viral shedding determination in 58 COVID-19 confirmed Iranian subjects in different stages. Methods. 58 COVID-19 confirmed Iranian subjects including 21 outpatients and 37 inpatients were investigated. The analytical data and clinical properties were documented in the standard questionnaire. The RT-PCR tests were done two and three weeks after the symptoms initiation. Results. Viral eradication occurred in 44.8% two weeks after illness initiation whereas in 71% who achieved a negative PCR test in the third week. Moreover, prolonged viral shedding was observed in hospitalized cases in comparison to outpatients. Almost 30% of patients continued viral shedding three weeks after disease initiation. Conclusion. A longer duration of viral shedding in hospitalized cases rather than outpatients was observed in this study. The results similar to other investigations call into question if the current policies are enough to prevent the viral spread or not. This study should be done on a larger sample to provide an appropriate time in isolation policy