The effect of tapered master gutta-percha cone on apical seal of straight and curved root canals prepared with nickel-titanium rotary files.

BACKGROUND: Gutta-percha has been the predominant root canal filling material which is developed with different taper. Canal obturation fixed with nickel-titanium (NiTi) instruments and tapered gutta-percha master cone and lateral condensation is advantageous because it is clinically effectual and appears to result in a radiographically acceptable outcome. The aim of this in vitro study was to determine the effect of tapered master gutta-percha cone on apical seal of straight and curved root canals using NiTi rotary files. MATERIALS AND METHODS: In this in vitro study total of 130 mandibular molars were selected and divided into six experimental groups (n = 20) based on the degree of root canal curvatures (0°-20°and 20°-40°) and the taper of master cones (0.02, 0.04, and 0.06). The roots were immersed in the bacterial leakage model and monitored daily for a period of 70 days. Data were analyzed using Kaplan-Meier approach, log-rank test, and Chi-square tests. P < 0.05 was considered statistically significant. RESULTS: The microleakage in the 0°-20° canal curvature using 0.02- and 0.04-tapered master cones was similar and considerably <0.06-tapered master cone (P < 0.05). However, the microleakage in the 20°-40° canal curvature using 0.02- and 0.04-tapered master cones was more than 0°-20° and for 0.06-tapered master cone was <0°-20°, but there was no statistical difference between the use of 0.02-, 0.04-, and 0.06-tapered master cones (P > 0.05). CONCLUSION: The lateral condensation filling technique using 0.02- and 0.04-tapered master cones is more effective in minimizing microbial leakage in straight canals than 0.06-tapered master cone.

Detection of Streptococcus gallolyticus in colorectal cancer and inflammatory bowel disease patients compared to control group in southwest of Iran.

There are several pieces of evidence regarding the role of bacteria, such as Streptococcus bovis/gallolyticus in the etiology of gastrointestinal diseases such as colorectal cancer (CRC) and inflammatory bowel disease (IBD). Therefore, the aim of this study was to detect S. gallolyticus subsp. gallolyticus (Sgg) in fecal samples of CRC and IBD patients by culture and molecular methods, in Ahvaz, southwest of Iran. A total of 106 fecal samples were collected from 22 CRC patients, 44 IBD patients, and 40 healthy individuals. The prevalence of Sgg was investigated by culture and polymerase chain reaction (PCR) with specific primers for sodA gene. The results of the stool culture showed that the overall prevalence of Sgg was 9 (13.6%) out of 66 patients. Meanwhile, the number of Sgg isolated from IBD and CRC patients was 7 (15.9%) and 2 (9%), respectively. The bacteria were not isolated from any of the control groups. On the basis of PCR, S. gallolyticus was detected in 24 (36.4%) out of 66 patients. Meanwhile, the number of IBD patients with positive sodA gene was 15 (34.1%) out of 44 cases. In CRC patients, the sodA gene was detected in 9 (40.9%) of 22 cases. Two (5%) of the specimens in the control group had the sodA gene. According to our results, S. gallolyticus subsp. gallolyticus might be involved in CRC and IBD pathogenesis. More investigation with different samples in the various areas might be shaded light on these results.

Isolation and characterization of Listeria monocytogenes from environmental and clinical sources by culture and PCR-RFLP methods.

BACKGROUND AND OBJECTIVES: Due to the widespread distribution of Listeria monocytogenes in environmental and animal sources and serious clinical complications in human, this study was aimed to isolate L. monocytogenes from water and clinical specimens by culture and PCR methods and to investigate the presence of hlyA and inlA virulence genes. MATERIALS AND METHODS: Water and clinical samples of vaginal and fecal were screened for the presence of L. monocytogenes by phenotypic and standard biochemical tests. PCR amplification was performed on extracted DNA using primers based on the hlyA and inlA genes. A 733-bp fragment of inlA gene was used for investigation of polymorphism using RFLP analysis. RESULTS: In total, 45 phenotypically and molecularly confirmed L. monocytogenes strains were isolated from different sources including 30 (16.7%) from water, 9 (11.3%) from vaginal swabs and 6 (7.5%) from fecal samples. RFLP analysis of PCR products using AluI and Tsp509I restriction enzymes, generated two profiles with 8 to 10 bands ranging in size from 15 to 210 bp. The majority of water and clinical isolates were classified in profile 2. CONCLUSION: We demonstrated 45 L. monocytogenes isolates from tested water and clinical samples by phenotypic and molecular tests. The majority of the isolates were classified in the same RFLP profile, showing the water as a potential source of clinical complications in patients in the region of study.

Genetic basis for metronidazole and clarithromycin resistance in Helicobacter pylori strains isolated from patients with gastroduodenal disorders.

BACKGROUND: The aim of this study was to evaluate the antimicrobial resistance and genetic basis for metronidazole (Mtz) and clarithromycin (Cla) resistance in strains of Helicobacter pylori, isolated from patients with gastroduodenal disorders. PATIENTS AND METHODS: A total of 157 H. pylori isolates (from 22 gastric cancer, 38 peptic ulcer disease, and 97 non-ulcer dyspepsia patients) were analyzed for drug susceptibility to Mtz and Cla, by gradient diffusion test (E-test, MAST). The PCR and sequence analysis of the rdxA and frxA for Mtz-resistant strains and the 23S rRNA for Cla-resistant strains were used to determine the genetic basis of drug resistance in H. pylori strains. Increased expression of TolC homologous genes (hefA) that upregulates efflux pump activity was determined in multidrug-resistant (MDR) strain of H. pylori by real-time PCR technique. RESULTS: Among 157 H. pylori isolates, 32 (20.4%) strains were resistant to at least one of the antimicrobial agents. The highest resistance rate was attributed to Mtz (n=69, 43.94%). Among the resistant strains of H. pylori, 15 cases (9.55%) were detected as MDR. Mutations in the rdxA (85.5%) and A2143G point mutations (63.1%) in the 23S rRNA were the most common cause of resistance to Mtz and Cla in strains of H. pylori, respectively. In MDR strains, the rdxA mutation and A2143G-point mutation in the 23S rRNA were the most abundant mutations responsible for drug resistance. The relative expression of hefA in MDR strains (mean 3.706) was higher than the susceptible strains (mean 1.07). CONCLUSION: Mutational inactivation and efflux pump overexpression are two mechanisms that increase the resistance to H. pylori antimicrobial agents and the rate of MDR strains. In Iran, the mutations of rdxA and frxA in Mtz-resistant strains and A2143G and A2142G of the 23S rRNA in Cla-resistant strains were significant. The screening for these mutations could help to prevent antibiotic resistance, and to determine the most effective anti-H. pylori drugs.

Prevalence and antimicrobial resistance of Shigella species isolated from diarrheal patients in Ahvaz, southwest Iran.

INTRODUCTION: Shigellosis is a significant global human health problem, and Shigella is in charge of almost 165 million cases of this disease annually, of whom 163 million cases are in developing countries and 1.5 million cases are in developed countries. The main aims of the current survey were to identify Shigella spp. isolated from diarrheal patients by conventional biochemical tests, determine the antimicrobial susceptibility profiles by disk diffusion method, and detect the ipaH gene using the PCR assay. METHODS: The bacterial isolates were identified as Shigella spp. by microbiological tests and were serogrouped by the slide agglutination test. Antimicrobial susceptibility testing was performed using the disk diffusion method. PCR was performed to detect the ipaH gene. RESULTS: The Shigella strains were isolated from 522 patients with various diarrhea, including bloody diarrhea (3%), mucoid plus bloody diarrhea (1.9%), mucoid diarrhea (3.2%), and watery diarrhea (3.2%). Overall, 69 (13.2%) isolates were positive for Shigella spp., of which 34 (49.3%) serotypes were identified as Shigella flexneri, 22 (31.9%) serotypes were identified as Shigella sonnei, 9 (13%) serotypes were identified as Shigella boydii, and 4 (5.8%) serotypes were identified as Shigella dysenteriae. Antibiotic susceptibility tests revealed that the highest resistance percentage was related to ampicillin (82%) and trimethoprim-sulfamethoxazole (77%), and ciprofloxacin and ceftriaxone were the best antibiotics against Shigella isolates. CONCLUSION: We concluded that Shigella spp. can be considered as an etiological agent of diarrhea in southwest Iran. Since the drug resistance pattern of Shigella differs geographically and over time within a country, continuous and regular surveillance program is necessary.

Prevalence of enterotoxin-encoding genes among diverse Shigella strains isolated from patients with diarrhea, southwest Iran.

Shigella spp. are a major cause of bacillary dysentery, particularly among children in developing countries such as Iran. This study aimed to investigate the presence of two important Shigella enterotoxins (ShET-1 and ShET-2), encoded by the set and sen genes, respectively, by polymerase chain reaction (PCR) assay among Shigella species isolated from children affected by shigellosis in Ahvaz, southwest of Iran. In this cross-sectional study, from June 2016 to April 2017, altogether 117 Shigella isolates were collected from fecal specimens of children aged <15 years with diarrhea in Ahvaz, southwest Iran. All isolates were identified by standard microbiological and molecular methods. The presence of enterotoxin genes was determined by PCR. The most prevalent isolate was Shigella flexneri (47.9%), followed by Shigella sonnei (41%) and Shigella boydii (11.1%), respectively. Shigella dysenteriae was not detected in patients' samples. The frequencies of set1A, set1B, and sen genes were 5.1% (6/117), 15.4% (18/117), and 76.9% (90/117), respectively. This study provides initial background on the prevalence and distribution of the Shigella enterotoxin genes in Shigella isolates in southwest of Iran. In addition, this study revealed a high prevalence of sen enterotoxin gene in Shigella species.

Evaluation of Antibiotic Susceptibility Pattern and Distribution of coa Genes in Coagulase-negative S. aureus from Ahvaz, Iran.

BACKGROUND: Staphylococcus aureus is one of the important causes of clinical infections that can be more destructive by its antibiotic resistant strains. OBJECTIVE: This study aimed to determine the antibiotic susceptibility pattern and distribution of mecA and coa genes in clinical isolates of S. aureus. METHODS: Two hundred seventy-three specimens suspected to S. aureus were taken from hospitals of Ahvaz, southwest of Iran. Isolates were identified by standard microbiologic tests and confirmed by the molecular method. Antimicrobial susceptibility testing was carried out by disk diffusion method. The presence of mecA and coa genes was determined by PCR method. RESULTS: Of a total of 200 isolates which were tested for coagulase tube test, 143 (71.5%) showed coagulase positive, and 57 (28.5%) showed a coagulase-negative reaction. Antibacterial susceptibility pattern of 200 S. aureus isolates showed the highest and lowest susceptibility rate to linezolid (98%) and ciprofloxacin (42%), respectively. The prevalence of methicillin-resistant S. aureus (MRSA) by detection of mecA gene was estimated as 47.5 % (95/200), of which the rate of MRSA in coagulase positive and negative isolates was 35% (50/143), and 65% (45/57), respectively. Meanwhile, coa gene was detected in 100% of coagulase positive and 28.1% of coagulasenegative isolates. CONCLUSION: The results of this study showed that the number of atypical CNSA in our area is high. Since the coagulase test is an essential test for diagnosis of S. aureus, our findings regarding the emergence of CNSA are a warning about the misdiagnosis and selection of appropriate treatment approach for S. aureus isolates.

CagA and vacA allelic combination of Helicobacter pylori in gastroduodenal disorders.

BACKGROUND: Allelic variation of the virulence genes, vacA and cagA, as the most important virulence associated genes play an important role in the pathogenesis of severe gastrointestinal disease. OBJECTIVE: The aim of the present study was to identify the diversity of the virulence genes in patients with Gastric Cancer (GC), who were referred to the gastro-endoscopy unit of Imam Khomeini Hospital, Ahvaz Jundishapur University of medical science, Ahvaz, Iran. METHODS: Gastric biopsy specimens from 301 patients suspected to gastrointestinal disorders, were analyzed for H. pylori using molecular and phenotypical methods (culture, and biochemical test (catalase, oxidase and urease tests)). RESULTS: Among 201 PCR positive for H. pylori, using histopathological methods, 22 (10.9%) patients had GC. Presence of vacA gene in our H. pylori strains was 100% (201/201), while the most virulent vacA s1 allele was detected in 82.6% isolates, and the mid region vacA m1 was found in 39.8% isolates. The vacA s1/m1 genotype was the most virulent allelic combination in GC and Peptic Ulcer Disease (PUD) in 68.2% and 50%, respectively. The cagA gene was detected in 66.7% isolates. Among the cagA positive isolates, EPIYA-ABC motif was the most common motif in the GC (66.7%), PUD (55.6%) and Erosive Gastroduodenitis (EG) samples (55.2%), while EPIYA-ABCC was the most common motif (58.7%) in the Non-Ulcer Dyspepsia (NUD) samples. The vacA s1m1/cagA+ combination was detected in GC (73.3%) and PUD (51.9%), while vacA s1m2/cagA+ presented in the NUD and EG samples in 77.8% and 62.1%, respectively. CONCLUSION: In this work, Western type (EPIYA-ABC and ABCC motifs) cagA, vacA s1m1 combinations have been demonstrated as the dominant genotype in the tested Ahvazian H. Pylori strains. Also the participation of cagA gene and vacA s1m1 genotype in development and severity of gastric disorder was well evident. Therefore, infection with H. pylori strain containing the cagA gene or the vacA s1m1 genotypes could be associated with increased risk of GC. This is the first study in our area that reports the high incidence and diversity of allelic combination of cagA and vacA genes in gastroduodenitis patients.

Pathogen Identification in Suspected Cases of Pyogenic Spondylodiscitis.

Pyogenic spinal infection continues to represent a worldwide problem. In approximately one-third of patients with pyogenic spondylodiscitis, the infectious agent is never identified. Of the cases that lead to organismal identification, bacteria are more commonly isolated from the spine rather than fungi and parasites. This study applied universal prokaryotic 16S rRNA PCR as a rapid diagnostic tool for the detection of bacterial agents in specimens from patients suspected of pyogenic spondylodiscitis. Gram and Ziehl-Neelsen staining were used as a preliminary screening measure for microbiologic evaluation of patient samples. PCR amplification targeting 16S rRNA gene was performed on DNA extracted from 57 cases including specimens from epidural abscesses, vertebral, and disc biopsies. Positive samples were directly sequenced. MRI findings demonstrated that disc destruction and inflammation were the major imaging features of suspected pyogenic spondylodiscitis cases, as 44 cases showed such features. The most common site of infection was the lumbar spine (66.7%), followed by thoracic spine (19%), the sacroiliac joint (9.5%), and lumbar-thoracic spine (4.8%) regions. A total of 21 samples amplified the 16S rRNA-PCR product. Sanger sequencing of the PCR products identified the following bacteriological agents: Mycobacterium tuberculosis (n = 9; 42.9%), Staphylococcus aureus (n = 6; 28.5%), Mycobacterium abscessus (n = 5; 23.8%), and Mycobacterium chelonae (n = 1; 4.8%). 36 samples displayed no visible 16S rRNA PCR signal, which suggested that non-bacterial infectious agents (e.g., fungi) or non-infectious processes (e.g., inflammatory, or neoplastic) may be responsible for some of these cases. The L3-L4 site (23.8%) was the most frequent site of infection. Single disc/vertebral infection were observed in 9 patients (42.85%), while 12 patients (57.15%) had 2 infected adjacent vertebrae. Elevated erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) inflammatory markers were noted in majority of the patients. In conclusion, microbiological methods and MRI findings are vital components for the proper diagnosis of pyogenic spondylodiscitis. Our findings suggest that molecular methods such as clinical application of 16S rRNA PCR and sequencing may be useful as adjunctive diagnostic tools for pyogenic spondylodiscitis. The rapid turnaround time of 16S rRNA PCR and sequencing submission and results can potentially decrease the time to diagnosis and improve the therapeutic management and outcome of these infections. Although S. aureus and M. tuberculosis were the most common causes of pyogenic spinal infections in this study, other infectious agents and non-infectious etiologies should be considered. Based on study results, we advise that antibiotic therapy should be initiated after a definitive etiological diagnosis.

Prevalence of Escherichia coli O157:H7 in Children with Bloody Diarrhea Referring to Abuzar Teaching Hospital, Ahvaz, Iran.

INTRODUCTION: Escherichia coli O157: H7 are recognized as important aetiological agents of diarrhea in children, particularly in developed countries. AIM: The aim of the study was to determine the rates of detection of E. coli O157: H7strains among children in Ahvaz, Iran. MATERIALS AND METHODS: From June 2010 to December 2010, 137 diarrheal stool samples of children were collected. E.coli was identified by standard microbiological techniques. O157 or O157:H7 subtypes discerned by serological tests. RESULTS: Of the 137 E. coli isolates, enteropathogens were found in 53 (38.7%) of the patients as follow: Shigella spp. (75.5%), EPEC (enteropathogenic E. coli) (16.9%), Campylobacter spp. (3.8%) and Salmonella spp. (3.8%). None of the isolated E. coli was O157:H7 serotype. CONCLUSION: This shows that non-O157:H7 E. coli are the major cause of paediatric infections in this region of Iran.