Genetic analysis of cagA and vacA genes in helicobacter pylori isolates and their relationship with gastroduodenal diseases in the west of Iran

Background: Helicobacter pylori have different virulence factors which are associated with several gastroduodenal diseases; however, this association is variable in different geographical regions. Data of genotypes of Iranian H. pylori isolates are few. Objectives: The aim of the current study was to investigate the cagA/vacA genotypes of Helicobacter pylori isolates and determine the relationship between these genotypes with respect to different gastric disorders in patients of Chaharmahalo Bakhtiarian. Materials and Methods: In this cross-sectional study, gastric biopsies were taken from 200 patients with gastrodoudenal diseases. Histopathological features were recognized by specialist. The samples were subjected to PCR for detection and identification of ureC, cagA and vacA genes. Results: The frequency of the vacA genotypes, sa1/m1, s1a/m1b, s1a/m2, s1b/m1a, s1b/m1b, s1b/m2, s1c/m1a, s1c/m1b, s1c/m2, s2/m1a, s2/m1b and s2/ m2 were 27(6.6%), 8(4.3%), 45(28.04%), 7(3.7%), 5(2.5%), 10 (6.1%), 12 (7.4%), 4 (2.5%), 18(11%), 6(3.7%), 0 and 22(13.5%) respectively. The cagA gene was detected in 92% of strains. Based on our findings, it seemed that cagPAI and vacA s1 genotypes were associated with some gastric disorders in patients with H. pylori. In this region, the isolates carrying s1a/m2 were the most prevalent. Conclusions: We found considerable relationship between s1a/m1a, s1a/m2, s2/m2 and s1c/m1a and some gastric disorders. Further studies about the role of H. pylori virulence factors and gastric disorders were recommended. © 2013, Iranian Red Crescent Medical Journal

Study of Helicobacter pylori genotype status in cows, sheep, goats and human beings

BACKGROUND: Helicobacter pylori is one of the most controversial bacteria in the world causing diverse gastrointestinal diseases. The transmission way of this bacterium still remains unknown. The possibility of zoonotic transmission of H. pylori has been suggested, but is not proven in nonprimate reservoirs. In the current survey, we investigate the presence of H. pylori in cow, sheep and goat stomach, determine the bacterium virulence factors and finally compare the human H. pylori virulence factors and animals in order to examine whether H. pylori might be transmitted from these animals to human beings. METHODS: This cross- sectional study was performed on 800 gastric biopsy specimens of cows, sheep, goats and human beings. The PCR assays was performed to detection of H. pylori, vacA and cagA genes. The PCR products of Ruminant’s samples with positive H. pylori were subjected to DNA sequencing analysis. Statistical tests were applied for data analysis. RESULTS: Overall 6 (3%) cows, 32 (16%) sheep and 164 (82%) human beings specimens were confirmed to be H. pylori positive; however we were not able to detect this bacterium in all 200 goat samples. The vacA s1a/m1a was the predominant H. pylori genotype in all three kinds of studied population. There was 3.4–8.4% variability and 92.9-98.5% homology between sheep and human samples. CONCLUSIONS: Considering the high sequence homology among DNA of H. pylori isolated from sheep and human, our data suggest that sheep may act as a reservoir for H. pylori and in the some extent share the ancestral host for the bacteria with human

Comparisons of competitive enzyme-linked immunosorbent assay and one step RT-PCR tests for the detection of Bluetongue virus in south west of Iran

Bluetongue is a noncontagious, arthropod-borne viral disease of both domestic and wild ruminants. Bluetongue virus (BTV) is the type of species of the genus Orbivirus within the family Reoviridae. BTV is endemic in some areas with cattle and wild ruminants serving as reservoirs for the virus. Clinical symptoms are often seen in sheep. There are several methods for the detection of Bluetongue virus, among them the molecular technique like RT-PCR is considered as the most sensitive and reliable one. The aim of this study was to comprise competitive enzyme-linked immunosorbent assay (C-ELISA) with one step RT-PCR test for the detection of BTV in sheep. A total of 770 blood samples were obtained from sheep (265 serum positive samples and 505 serum negative samples in C-ELISA). According to our data, out of the 265 serum positive samples in ELISA test, 234 were positive in RT-PCR assay whereas all serum negative samples were negative in RT-PCR experiment. According to the results, the PCR assay was more sensitive and reliable than ELISA technique for the diagnosis of Bluetongue virus.Key words: Bluetongue virus, C-ELISA, RT- PCR, Sheep, Iran

PP-005 Clarithromycin resistance assessment in Helicobacter pylori isolates by using 23S rRNA gene molecular markers

Background: H. pylori is a relatively fastidious and microaerophilic microorganism and therefore standard phenotypic susceptibility tests, even in the hands of experts, are slow and can take at least 10 14 days. Molecular based diagnostic assays by using molecular markers for resistance detection offer an attractive alternative approach to obtain susceptibilities to antibiotics with greater accuracy and speed, and the possibility of a same day result. The aim of this study is the assessment of clarithromycin resistance by using molecular markers. Methods: This cross-sectional descriptive study was performed on 200 gastric biopsy specimens which were obtained from patients undergoing upper gastrointestinal tract endoscopy in Hajar hospital of Shahrekord, by using TaqMan real-time PCR. Initially, H. pylori strains were identified by RUT and PCR. Then, by this regard that accumulation of mutations associated with resistance to clarithromycin were in the region between nucleotides 2142 2144 of 23S rRNA gene, the first probe was designed to be able the distinguish between sensitive and resistant strains. Finally four probes were designed that each be able to identify only one mutation associated with a particular level of clarithromycin resistance. Results: Out of 200 samples, 164 (82%) were H. pylori positive. Overall, clarithromycin susceptible strains were detected in 105 (64.02%) patients and clarithromycin resistance were detected in 59 (35.98%) which were identified as 4 (2.44%) A2144G, 26 (15.85%) A2143G, 15 (9.15%) A2143C and 20 (12.19%) A2142G point mutations. Purely resistant strains were detected in 38 (23.17%), while heteroresistant were found in the remaining 16 (9.76%) cases. Genotype of 5 (8.47%) strains was not detected. This data was confirmed by PCR-RFLP technique. Conclusion: Results showed that Real-time PCR assay in combination with molecular markers has high accuracy to simultaneously identify H. pylori and clarithromycin resistance types directly in gastric biopsy specimens in short time

Uropathogenic Escherichia coli in Iran. Serogroup distributions, virulence factors and antimicrobial resistance properties

Background: Urinary tract infections (UTIs) are one of the most common bacterial infections with global expansion. These infections are predominantly caused by uropathogenic Escherichia coli (UPEC).Methods: Totally, 123 strains of Escherichia coli isolated from UTIs patients, using bacterial culture method were subjected to polymerase chain reactions for detection of various O- serogroups, some urovirulence factors, antibiotic resistance genes and resistance to 13 different antibiotics.Results: According to data, the distribution of O1, O2, O6, O7 and O16 serogroups were 2.43%, besides O22, O75 and O83 serogroups were 1.62%. Furthermore, the distribution of O4, O8, O15, O21 and O25 serogroups were 5.69%, 3.25%, 21.13%, 4.06% and 26.01%, respectively. Overall, the fim virulence gene had the highest (86.17%) while the usp virulence gene had the lowest distributions of virulence genes in UPEC strains isolated from UTIs patients. The vat and sen virulence genes were not detected in any UPEC strains. Totally, aadA1 (52.84%), and qnr (46.34%) were the most prevalent antibiotic resistance genes while the distribution of cat1 (15.44%), cmlA (15.44%) and dfrA1 (21.95%) were the least. Resistance to penicillin (100%) and tetracycline (73.98%) had the highest while resistance to nitrofurantoin (5.69%) and trimethoprim (16.26%) had the lowest frequencies.Conclusions: This study indicated that the UPEC strains which harbored the high numbers of virulence and antibiotic resistance genes had the high ability to cause diseases that are resistant to most antibiotics. In the current situation, it seems that the administration of penicillin and tetracycline for the treatment of UTIs is vain

The Introduction of Audit Committees and Implementation of The Corporate Governance Code in Kuwait

This thesis explores the introduction of audit committees (ACs) and implementation of the corporate governance code (CGC) in Kuwait. The study has five aims: (1) to investigate the reasons for the delay in issuing the ACs’ regulations before the release of the CGC in 2013; (2) to explore why some companies voluntarily formed an AC while others did not; (3) to understand how listed companies perceived, experienced and responded to the CGC’s introduction in 2013 and the reasons for their reactions; (4) to analyse the strategic responses that companies used when the mandatory code was introduced, which led to changing the code; and (5) to explore the interviewees’ perceptions regarding the extent to which they believe that listed companies will implement the AC effectively. The data were collected using semi-structured interviews with key individuals. The institutional logics perspective was used as the theoretical framework to analyse and interpret the study’s findings. The results show that the main obstacle to implementation of CG and ACs in Kuwait is the dominance of family logic. Moreover, the reasons for the delay in issuing the regulations for ACs may be directly or indirectly due to the domination of market and family logic over state logic. The findings indicate that family, market, state, corporate and professional logics have most influence on the decision to voluntarily form ACs. The findings also reveal that the companies guided by state and corporate logic perceived the CGC as compatible with institutional demands, while the companies embedded in family and market logic perceived it as a conflicting demand. The companies and other interested parties used compromise, defiance and manipulation as a response to the introduction of the mandatory code. Most interviewees believed that companies will implement the ACs symbolically because of the dominance of family logic in Kuwait

PURIFICATION AND CHARACTERIZATION OF PROTEASE FROM Pseudomonas aeruginosa ISOLATED FROM SOME WOUND AND BURN INFECTION.

Twenty Four isolates of Pseudomonas aeruginosa were identified. The isolates were 8(33%) from wound infections, 16(66%) isolates from burn infections. The sensitivity of Pseudomonas aeruginosa isolates was been tested against (10) antibiotics showed isolates version resistance with different percentage against antibiotics. Pseudomonas aeruginosa exhibited (100%) resistance to Ampicillin. While percentages of resistance to Cefixime and Ceftazidime were (95.8%) and (79%) respectively. Resistance percentages to Tobramycin, Piperacillin, Norfloxacillin, Ciprofloxacin were (41.6%), (20.8%), (20.8%) and (4%) respectively. All isolates of Pseudomonas aeruginosa were highly sensitive (100%) to Aztronam, Imipenem, Cefepime. The optimum conditions for protease production were in LB medium with a pH (8) after (48) hrs of incubation at (35) Cº. Purification of the protease was done using ion exchange chromatography DEAE-cellulose and gel filtration with sephadex G-100. Molecular weight of the purified protease was measured by sephadex G-100 and it was found to be around (21379) Dalton. The optimum temperature of enzyme activity was (35) Cº. However, the pH (8) was for activity and stability of this enzyme. Zn++ and Ca++ ions may play a role in the enhancement and stability of the enzyme. Enzyme activity was not inhibited in the presence of reducing agent such as Cysteine, but it was inhibited in the presence of EDT

Study of Helicobacter pylori genotype status in saliva, dental plaques, stool and gastric biopsy samples

AIM: To compare genotype of Helicobacter pylori (H. pylori) isolated from saliva, dental plaques, gastric biopsy, and stool of each patient in order to evaluate the mode of transmission of H. pylori infection