[Frequency of sialic acid binding adhesin gene in Helicobacter pylori isolated from patient with gastroduodenal diseases]

Sialic acid binding adhesin gene is one of the most important factors contributing to adhesin of Helicobacter pylori [H. pylori] to epithelial cell layer of stomach. The prevalence rates of sialic acid binding adhesin gene vary in different geographic areas. The aim of this study was to determine the frequency of sialic acid binding adhesin coding gene in the patients with different gastroduodenal diseases. This is a cross-sectional study. One hundred twenty patients with GI symptoms were enrolled in this study. Two gastric biopsy specimens were taken from each of the patients for rapid urease test [RUT] and DNA extraction. Presence of H. pylori was investigated by RUT and urease A gene [ureA] PCR. sialic acid binding adhesin gene was detected by using gene specific primers. Among 120 samples, presence of H. pylori was confirmed in 82 cases, of which 64 strains [78%] were positive for sialic acid binding adhesin gene. The frequency of this gene was 84.6%, 86.7%, 77.8% and 72.2% for gartric cancer, gastric ulcer, duodenal ulcer and gastritis [%66.7] respectively. The frequency of sialic acid binding adhesin gene in different samples was almost the same. Discrepancies in the frequency of this gene in different studies may be related to geographical diversity or use of different primers for detection of this gene

frequency of extended spectrum beta lactamase and CTX M-I of Escherichia coli isolated from the urine tract infection of patients by phenotypic and PCR methods in the city of Khoy in Iran

The production of Extended Spectrum Beta Lactamases [ESBLs] by Escherichia coli is the main cause of resistance to Cephalosporins. In the past decade, CTX-M enzymes have become the most prevalent ESBLs in Europe, Canada, and Asia. In this study, the frequency of ESBL- producing E.coli and molecular detection of the CTX-M-I group was investigated. A total of 400 urine samples were collected from both hospitalized and out-patients in Khoy's hospitals between November 2009 and April 2010. Out of these samples, 188 were identified as E.coli by standard biochemical tests. The antibiotic Susceptibility tests to 10 antibiotics were performed by the-disk-agar diffusion [DAD] method. ESBL production was screened by phenotypic test that including disk diffusion agar and combined disk as recommended by the Clinical and Laboratory Standards Institute [CLSI] Screened isolates were investigated by PCR assay for detection of CTX-M-I group genes. The results show that out of 188 E.coli isolates identified, 56 [29.8%] were producing ESBls by phenotypic test. All isolates were sensitive to imipenem. Overall, 49 [87.5%] isolates were confirmed as CTX-M-I producer by PCR. The results of this study showed that about 30% of the identified E.coli were producing ESBL Therefore, we recommend to use molecular methods in such researches

[Study of antibacterial properties of adiantum capillus-veneris extract on eight species of gram positive and negative bacteria]

Adiantum capillus-veneris L. is a traditional medicinal plant which was used in the treatment of bronchitis and coughs, and also to prevent hair loss. Methanolic extract of this plant has demonstrated significant antimicrobial activities. In this study the antibacterial properties of Adiantum capillus-veneris L. extract on eight species of Gram positive and negative bacteria were evaluated. The herbal sample of Adiantum capillus-veneris was collected during the summer [June-July] from the north region of Iran called Condoluse and identified by herbarium laboratory in Faculty of Pharmacy, Tehran University of Medical Sciences, where a voucher specimen is deposited. The sample was pretreated and extracted with methanol 96% by percolation method and then concentrated and stored in a safe bottle until the experiments started. By using dilution method different dilutions of the extract have been prepared [10%, 5%, 2.5% and 1.25%]. Antimicrobial activity of the methnolic extract of Adiantum capillus-veneris were evaluated against 8 strain of Gram positive and Gram negative bacteria using agar disk diffusion and agar well plate methods. Our results demonstrated that prepared dilutions of the Adiantum capillus-veneris extract had significant effects on Staphylococcus aureus, Escherichia coli and Helicobacter pylori strains. Also the results indicated no significant inhibitory effects on Salmonella typhi, Shigella sonnei, Pseudomonas aeroginosa, Proteus vulgaris and Streptococcus pyogenes strains. Consistent with the other studies, our investigation demonstrated some antimicrobial effects of Adiantum capillus-veneris extract. With respect to various and multiple chemical properties of this plant, it is suggested that Adiantum capillus-veneris can be used for more medical and therapeutic purposes

Simultaneous detection of caga and cage of helicobacter pylori strains recovered from Iranian patients with different gastroduodenal diseases

To asses the status of two representative genes of cag PAl i.e cagA and cagE of Helicobacter pylori strains infecting Iranian patients suffered from various clinical outcomes using one-step PCR. A total of 120 H. pylori infected patients including non-ulcer dyspepsia, NUD [n=81], peptic ulcer disease, PUD [n=17], and gastric carcinoma, GC [n=22] referred for endoscopy or gastric resection to Amir Alam Hospital or Cancer Institute from 2005 to 2008 were assessed. The status of cagA and cagE genes was determined by gene specific PCR. 84.2% and 90.8% of the tested strains were positive for cagA and cage, respectively. 81.7% strains were positive for both cagA and cagE genes, whereas 8 [6.7%] were found double negative. The prevalence of cagA in GC patients [100%] was slightly higher than PUD patients [94.1%]. All of GC cases were infected with cagA-positive strains. The same distribution pattern was indicated for cagE gene in GC and PUD patients. The cagA-positive strains were significantly associated with GC as compared with NUD [P< 0.05] but this association did not gain statistical significance when cagE gene was assessed. The concurrent detection of cagA/cagE genes allowed rapid and specific clarification of cag PAI status. The strains with cagA/cagE genotype are predominant in Iran regardless of clinical outcome and create a distinct cluster pattern from those in the West and similar to those of East Asian countries. The current study also demonstrated that cagE gene can be explored as a better indication of cag-PAI in Iranian H. pylori strains

Frequency of Helicobacter pylori vacA genotypes in Iranian patients with gastric and duodenal ulcer

Helicobacter pylori infection is a risk factor for developing chronic peptic ulcers and gastric cancer. The purpose of this study was to investigate the frequency of Helicobacter pylori vacA genotypes in patients with gastric and duodenal ulcer. A total of 100 biopsy specimens of patients with gastric [n = 50] and duodenal [n = 50] ulcer were collected. The specimens were cultured on selective media and incubated in a microaerophilic atmosphere at 37°C for 5-10 days. The isolates were characterized to species level by conventional biochemical tests. The extracted DNA from isolates was used to perform a polymerase chain reaction based, simultaneous analysis of the cagA status, allelic variation of the signal regions [s1, s2] and the middle regions [m1, m2] of the vacA gene. H. pylori isolated from 50 specimens of patients and the vacA gene was detected in all isolates. Among vacA genotypes the s1/m1 was the most common in H. pylori isolates from patients with gastric ulcer [56%] and duodenal ulcer [68%]. This study demonstrated that vacA slml is common genotype of H. pylori in patients with peptic ulcer and the vacA allele s1 of this bacterium is associated with ulcer

[Cloning and Sequencing of sacol a novel gene from Staphylococcus aureus]

Natural staphylococcal infections and vaccines based whole bacteria lead to poor antibody responses, but recent research reveals that specific antibodies based on recombinant staphylococcal antigens are much more protective. Sacol is a novel antigen that its structural and immunological traits poorly characterized. This research aimed to clone of sacol, a novel gene from Staphylococcus aureus. The specific primers with suitable restriction sites were designed and sacol amplified by PCR. The sacol and plasmid were produced as sticky ends by restriction enzymes NdeI and XhoI. To amplify the recombinant plasmid the pET21 sacol transferred into competent cell E.coliTOP10. The recombinant plasmid harvested from the host and analyzed by restriction enzymes and sequencing. Finally, sacol gene analyzed by bioinformatics tools. The sacol gene has 723bp which amplified, cloned and sequenced successfully. Sacol is highly conserved in Staphylococcus aureus strains. Moreover, software analysis shows that sacol encodes a protein with 32KDa molecular weight [267 amino acids] which has similarity with C51 peptidase in N-terminal with one alpha helix and 14 beta sheets. The sacol gene is conserved in majority of Staphylococcus aureus strains and may exist and express in most of staphylococcal infections. The role and regulation of the gene is thus of great interest

[Assessment the relationship of cagA gene with different gastroduodenal diseases in Helicobacter pylori infected patients]

The gastric pathogen Helicobacter pylori is introduced as an etiologic agent of gastritis and peptic ulcer and is associated with development of gastric adenocarcinoma. One of the most studied virulence marker of H. pylori is cytotoxin-associated gene A [cagA] with significant geographical heterogeneity around the world. This study was undertaken to assess the status of cagA gene of H .pylori strains infecting Iranian patients suffering from various gastrointestinal diseases and to evaluate the detection of this gene as a screening marker of high-risk patients. In this study, 180 patients [Mean age: 44 years] with upper gastrointestinal manifestations referred for endoscopy to Amir-Alam Hospital or Cancer Institute in Tehran were included. Among one hundred twenty H. pylori infected patients 81, 17 and 22 had non-ulcer dyspepsia [NUD], peptic ulcer disease [PUD], and gastric carcinoma [GC] respectively. Tissue samples were homogenized and incubation was performed up to 5 days. Identification was based on morphology under Gram staining and biochemical tests. The status of conserved region of cagA gene was determined by gene specific PCR. For statistical analysis, chi square test was used. Among the 180 of studied patients, 120 H. pylori strains were isolated. One hundred and one [84.2%] of the tested strains were positive for cagA and the remaining strains [15.8%] were negative. All of gastric cancer cases were infected with cagA -positive strains. The cagA -positive strains were significantly associated with GC as compared with NUD [p < 0.05] but this association did not gain statistical significance for other clinical outcomes. Although the possession of cagA is associated with GC when compared to NUD, due to the uniform distribution of cagA in all other disease categories detection of cagA alone can not be considered as a discriminative marker for a specific clinical outcome. Hence, the study of other virulence determinants and functional characteristics of cagA gene might be necessary for screening high risk patients

Prevalence of helicobacter pylori Cag E genotypes, in patients with non-ulcer dyspepsia, peptic ulcer and gastric carcinoma

Helicobacter pylori are a bacterial pathogen evolved to chronically colonize the gastric epithelium and causes gastritis, peptic ulcers, and even gastric malignancies in few infected humans. More recently, a pathogenicity island has been identified within the H. pylori genome that contains a cluster of genes, including cagE. The aim of the current study was to investigate the prevalence of cagE genotypes of H. pylori isolates from patients with NUD[Non Ulcer Dyspepsia], peptic ulcer and cancer. 150 Gastric biopsy specimens obtained from patients were inoculated onto a Brucella Columbia Agar containing 5% sheep for 3 to 5 days at 37°C under micro aerobic conditions [5% O2, 5% CO2, and 90% N2]. After DNA extraction, genotyping of the cagE gene was performed by PCR amplification using the primers. PCR products were separated by 1% agarose gel electrophoresis and examined under UV illumination. Of 92 positive cultures, 34, 28, 20, and 10 isolations were obtained from patients with NUD, duodenal ulcers [DU], gastric ulcers [GU] and gastric cancer [GC], respectively. The frequency of cagE gene was 88/24%, 100%, 85%, 100% and within isolates of patients with NUD, DU, GU and GC, respectively. The presence of cagE in patients with Helicobacter pylori infection is not a marker for predicting or diagnosing the resultant diseases

Genotyping of Helicobacter pylori strains isolated from patients with NUD, DU, GU and GC by RAPD-PCR

Helicobacter pylori is a genetically diverse gastric pathogen that chronically infects billions of people worldwide, typically beginning in infancy and lasting for decades. It is a major cause of peptic ulcers and it is an early risk factor for gastric cancer which is the most frequently lethal malignancy globally. This project was designed to genotype H. pylori isolates isolated from patients with NUD, DU, GU and GC by the polymerase chain reaction [PCR]-based on Randomly Amplified Polymorphic DNA [RAPD] fingerprinting technique. Eighty patients admitted to the gastroenterology unit at Sharyati hospital in Iran were included in this study. Gastric biopsy specimens were inoculated onto selective medium then were cultured for 3 to 5 days at 37 °C under microaerobic conditions. Genomic DNA was extracted using a commercially available Qiagen kit. RAPD-PCR was used to genotype isolates. Six different RAPD patterns [A-F] were seen in more than one isolate which were as follow; pattern A: 9 [16.98%], B: 6 [11.33%], C: 5 [9.43%], D: 3 [5.66%], E: 2 [3.77%] and F: 2 [3.77%]. Twenty six [49.06%] of 53 isolates showed a unique RAPD pattern that were not similar to each other. A significant relationship was not seen between a single RAPD pattern and a gastric disorder [P>0.05]. The results of this study suggest a high level of DNA sequence diversity among H. pylori isolates and it is better to use sequencing method for surveying of Helicobacter pylori genome rather than RAPD-PCR

Bacterial infections in renal transplant recipients

Urinary tract infection [UTI] is considered as one of the important bacterial infections seen among renal transplant recipients. In the present study, bacterial urinary tract infections in renal transplant recipients were investigated. Eighty-seven renal transplant recipients [57 males and 30 females] were included to study the bacterial UTIs. Clean- catch midstream urine specimens were obtained from patients and studied using microscopic analysis and culturing on appropriate bacteriologic media. Bacterial isolates were identified by standard biochemical and serological tests. UTIs were diagnosed in 29 percent of patients [18 males and 11 females]. The most common causative bacterial strains were coagulase negative Staphylococci [31%] and Entrobacter spp [20.7%]. The results showed that all of Proteus spp, Pseudomonas spp, Klebsiella spp, and Enterococcus spp were resistant to most of tested antibiotics, so this research reflects that these multiple resistant bacteria can be accounted as the most cause of UTI in renal transplant recipients

Prevalence of clostridium difficile- associated diarrhean in hospitalized patients with nosocomial diarrhea

Clostridium difficile is a frequently identified cause of nosocomial gastrointestinal disease. It has been proved to be a causative agent in antibiotic-associated diarrhea, antibiotic-associated colitis, and pseudomembraneous colitis. This study was aimed to determine the prevalence of C.difficile- associated diarrhea in hospitalized patients with nosocomial diarrhea. The 942 hospitalized patients stool samples with nosocomial diarrhea were collected at three hospitals in Tehran from Dec 2002 to Feb 2004.All the stool samples were cultured and in 97 [prevalence: 10.9%] samples grew C.difficile that 57 [prevalence: 6.1%] isolates were toxigenic by cytotoxicity assay and so 57 patients had C.difficile- associated diarrhea. Results of statistical analysis showed significant difference between the rate of C.difficile associated diarrhea and the patients ages [P<0.05]

Study of tuberculous infection rate in townships in a Centeral Province of Iran

Tuberculosis is a continuous threat for health in all parts of the world. An estimated one-third of the world' s population is infected with the Mycobacterium tuberculosis and 7 to 8 million people develop TB disease each year. The purpose of this study was to investigate the rate of tuberculosis in the townships of Yazd province, Iran. During the study period [19971999], 3885 suspected tuberculosis patients [1820 males and 2065 females; aged 8-85 years] who had been referred to the Yazd referral polyclinic were investigated by Ziehl Neelsen staining and culture method and questionnaire was completed for each subject. Then, Collected data were analyzed by statistical package for social science [SPSS] and chi-square program. The results show that, of the total suspected tuberculosis, 604 cases were found to be positive for tuberculosis. The average annual rate of tuberculosis was 26.8 cases per 100000 population [23.1/100000 males and 31/100000 females]. The highest and lowest rates of tuberculosis were observed among Sadough [78.1/100000] and Abarkouh townships population [19.8/100000] and also among age group >/= 50 years old [111/100000] and < 10 years old [7/100000], respectively. The average annual rates of pulmonary and extra-pulmonary tuberculosis in Yazd province were 152 cases [20.2%] and 48 cases [6.4%], respectively. It seems that, despite the efforts, which have been done for prevention, diagnosis and treatment of patients with tuberculosis, it is still considered as a threat for health in the Yazd province, Iran