Identification of Helicobacter species by 16S rDNA PCR and sequence analysis in human liver samples from patients with various etiologies of benign liver diseases.

BACKGROUND/AIMS: Several reports indicated an increased prevalence of the Helicobacter species in hepatocellular cancer tissue and in liver samples infected with hepatitis viruses. The frequency of Helicobacter spp. in benign liver diseases was, however, not thoroughly investigated. METHODS: Seventy-five consecutive patients with suspected liver disease were enrolled. The indications were hepatitis B virus (n=30), C virus (n=8), B and C dual infection (n=1), nonalcoholic steatohepatitis (n=27), autoimmune hepatitis (n=3), primary biliary cirrhosis (n=1) and idiopathic elevation of liver enzymes (n=5). PCR detection of 16S recombinant RNA gene of Helicobacter spp. was performed on liver samples. PCR products of positive samples were further identified by DNA sequencing. The patients also had upper gastrointestinal endoscopy and gastric biopsy for the detection of H. pylori using histopathology and PCR. RESULTS: Helicobacter spp. DNA was detected in two out of 75 liver biopsy samples (2.6%), which were typed as H. pylori by DNA sequencing. One of these patients had chronic hepatitis C infection (man, 51 years old) and the other had nonalcoholic steatohepatitis (woman, 44 years old). Fifty-two out of 75 of the patients (69.3%) had H. pylori infection in their stomachs. CONCLUSION: We have found that H. pylori infection is much less prevalent in benign liver diseases. The presence of H. pylori in nonalcoholic steatohepatitis (NASH) patients is a novel finding and this finding should be confirmed in a larger series.

Detection of Helicobacter pylori DNA in aortic and left internal mammary artery biopsies.

We investigated the relationship between acute coronary ischemia and the presence of Helicobacter pylori DNA in aortic regions that were absent macroscopic atheromatous plaques. The study group (Group 1) consisted of 42 patients who underwent coronary artery bypass grafting. Biopsy samples were obtained from 2 different locations: from regions of the aorta that were free (macroscopically) of atheromatous plaque (Group 1A), and from the internal mammary artery (Group 1B). The control group (Group 2) of 10 patients who had no atherosclerotic vascular disease provided aortic tissue samples for comparison. The real-time polymerase chain reaction method was used to detect H. pylori DNA in all biopsy samples. Eleven of 42 aortic tissue samples (26%) in Group 1A were positive for H. pylori DNA. Neither biopsies from the left internal mammary arteries of those patients nor biopsies from the aortas of the control group (Group 2) were positive for H. pylori DNA. There was a statistically significant difference between 1A and 1B in terms of H. pylori positivity (P=0.001). In Group 1 as a whole, acute coronary ischemia was more prevalent in the H. pylori-positive patients than in the H. pylori-negative patients (P=0.001). To our knowledge, this is the 1st study to investigate the detection of H. pylori DNA in aortic biopsy samples that are macroscopically free of atheromatous plaque. Such detection in patients who have atherosclerotic coronary artery disease could be an important indication of the role of microorganisms in the pathogenesis of atherosclerosis.

Effects of atypical pneumonia agents on progression of atherosclerosis and acute coronary syndrome.

BACKGROUND: The aim of this study was to investigate the presence of various atypical pneumonia agents (Chlamydia pneumoniae, cytomegalovirus, Mycoplasma pneumoniae), which are considered to have a role in the ethiopathogenesis of atherosclerosis, in aortic biopsies without macroscopically visible plaque and in internal thoracic artery biopsies. MATERIAL AND METHODS: Thirty-three patients (group 1), who had undergone coronary bypass operation and 10 non-atherosclerotic patients (group 2), were included in the study. Seventy-six tissue biopsies were taken. Biopsies from the patients in group 1 a were obtained from the atheroma plaque-free aortic tissue and 33 biopsies (group Ib) were obtained from their internal thoracic arteries. Following DNA extraction, nested PCR was used to detect Chlamydia pneumoniae DNA, and real time PCR was used to detect cytomegalovirus and Mycoplasma pneumoniae DNA. Blood parameters (lipid profile, CRP, fibrinogen) of the patients and operation characteristics were recorded. RESULTS: Chlamydia pneumoniae DNA was detected in 5 of 33 biopsy samples from coronary bypass patients, whereas none of the control patients (group 1b and group 2) were positive for this agent (P = 0.001). Neither CMV nor Mycoplasma pneumoniae was detected in IMA and aortic biopsies of both bypass and control patients. Elevated total cholesterol levels (P = 0.02) and positive CRP (P = 0.001) was found in C. pneumoniae positive patients. Prevalence of acute coronary syndrome was significantly higher in C. pneumoniae detected patients compared (P = 0.00 1). CONCLUSIONS: Detection of C. pneumoniae DNA in the atheroma free aortic biopsies might indicate that this micro-organism intervened in the progression of atheroma plaque. There was a strong relationship between the detection of this micro-organism in the aortic wall and acute coronary syndrome. The absence of DNA of the corresponding micro-organisms in the IMA wall may show its resistance to infective agents and in turn to atherosclerosis, which is a result of the prevailing endothelial functions of this artery.

A possible role of Helicobacter pylori in chronic rhinosinusitis: a preliminary report.

OBJECTIVE: The objective was to investigate the presence of Helicobacter pylori with polymerase chain reaction in the sinonasal mucosa of patients with or without chronic rhinosinusitis (CRS). STUDY DESIGN: A prospective clinical trial. METHODS: Mucosal tissue samples were collected from ethmoid cells of 12 patients with CRS and the removed mucosal part of the middle concha of 13 patients with concha bullosa who were treated surgically in our institution. DNA extracted from these samples was used for the amplification of 16S ribosomal RNA gene of H pylori by nested polymerase chain reaction. RESULTS: Helicobacter pylori DNA was detected in 4 of 12 patients with CRS, but it was not detected in patients with concha bullosa. Three of four patients with positive results for H pylori had gastroesophageal reflux-related complaints. CONCLUSIONS: It is possible to detect H pylori in the sinus mucosa of some patients with CRS. However, whether H pylori is a causative agent for CRS or a result of CRS is not known.

Detection of Helicobacter pylori and its CagA gene in tonsil and adenoid tissues by PCR.

OBJECTIVE: To determine the presence of Helicobacter pylori and, if detected, the prevalence of the CagA gene in adenotonsillectomy specimens by polymerase chain reaction (PCR). DESIGN: A prospective clinical trial. SETTING: Tertiary referral center. PATIENTS AND METHODS: The study population comprised 23 patients who had undergone adenoidectomy, tonsillectomy, or adenotonsillectomy under local or general anesthesia. Helicobacter pylori DNA was extracted from 3-mm-diameter tissue samples obtained from each tonsil and adenoid tissue specimens. The amplifications were performed for the 16S ribosomal RNA (rRNA) and CagA genes of H pylori in the samples of which H pylori DNA was detected. RESULTS: In examining all the samples, 7 (30%) of 23 patients were shown to be positive for H pylori DNA, 5 (71%) of whom also possessed the CagA gene. CONCLUSIONS: Tonsil and adenoid tissues may be an ecological niche of the mouth without regard to transient or permanent colonization. Oral-oral transmission may be a possible mode of spread of H pylori.

[Detection of cytomegalovirus DNA in immunosuppressed patients using qualitative and quantitative molecular methods]. / Immünsüpresif hastalarda sitomegalovirus DNA'sinin kalitatif ve kantitatif moleküler yöntemlerle arastirilmasi.

In this study, blood samples collected from 101 immunosuppressive patients were investigated for the presence of cytomegalovirus (CMV) DNA, with qualitative nested-polymerase chain reaction (PCR), and leukocytes obtained from these samples with quantitative hybrid capture assay (HCA). CMV-DNA was found positive in 32 (31.7%) and negative in 45 (44.5%) patients with both of the methods, and the agreement between the methods were estimated as 76.2%. The number of samples, which were PCR positive and HCA negative, were 24 (23.7%), while there were no samples which were PCR negative and HCA positive. All of the 56 CMV-DNA positive patients detected by PCR, were found positive for CMV-IgG, and 7 of them were also CMV-IgM positive. As a result, it was concluded that PCR is a practical and reliable method especially for the routine procedures for the investigation of CMV-DNA, however in cases which necessitate the detection of viral load, hybridization may be the preferable method.

The role of viral agents in aetiopathogenesis of acute rheumatic fever.

The reason why abnormal immune response exists in acute rheumatic fever is not exactly explained. The influence of co-pathogens like certain viruses were mentioned regarding the initiation of the immunological reaction in acute rheumatic fever patients by several authors since 1970. This study was designed to find the role or effect of some viral infections in the development of rheumatic fever. In this study, 47 cases with acute rheumatic fever (acute rheumatic arthritis, acute rheumatic carditis, and chorea), 20 cases with chronic rheumatic fever, 20 cases with streptococcal pharyngitis, and 20 healthy age- and gender-matched control cases were involved. Serological and molecular tests were made including hepatitis B virus, hepatitis C virus, rubella virus, herpes simplex virus (HSV group 1), and Epstein-Barr virus (EBV). HBsAg, rubella IgM and EBV IgM positivity were not seen in any of patients with rheumatic fever. Although antiHBs seropositivity was higher in the control group, it was not statistically significant (p > 0.05). There was no difference in rubella IgG, HSV IgM seropositivity, either (p > 0.05). EBV DNA was searched by the polymerase chain reaction technique; due to the latent nature of the virus, no significant difference was found between the control group and the other groups (p > 0.05). In this study, no positive correlation could be found to support the synergism theories regarding the streptoccocus infection and viral infections in the development of acute rheumatic fever. Only EBV DNA positivity was found in all acute rheumatic fever cases but not in the control group may lead to further studies with larger series of patients.

A follow up study of cytomegalovirus infection in a group of Turkish renal transplant recipients using molecular assays.

The clinical value of an in-house cytomegalovirus nested polymerase chain reaction (CMV-PCR) and a commercial molecular assay hybrid capture CMV DNA assay (HCA) was evaluated in monitoring a group of renal transplant patients for six months follow up. In this study, the sensitivity, specificity, positive predictive value, and negative predictive value of nested CMV DNA PCR assay and HCA at the beginning of the study were 70, 42.9, 46.7, 66.7, and 60, 78.6, 66.7, and 73.3% respectively. After six months, they were 80, 66.7, 80, 66.7 for CMV PCR and 73.3, 88.9, 91.7, 66.7% for HCA respectively. These results indicate that in monitoring and predicting CMV infections in renal transplant recipients, not only qualitative but also quantitative assays must be used together in order to decide the preemptive strategies.

A follow up study of cytomegalovirus infection in a group of Turkish renal transplant recipients using molecular assays

The clinical value of an in-house cytomegalovirus nested polymerase chain reaction (CMV-PCR) and a commercial molecular assay hybrid capture CMV DNA assay (HCA) was evaluated in monitoring a group of renal transplant patients for six months follow up. In this study, the sensitivity, specificity, positive predictive value, and negative predictive value of nested CMV DNA PCR assay and HCA at the beginning of the study were 70, 42.9, 46.7, 66.7, and 60, 78.6, 66.7, and 73.3 percent respectively. After six months, they were 80, 66.7, 80, 66.7 for CMV PCR and 73.3, 88.9, 91.7, 66.7 percent for HCA respectively. These results indicate that in monitoring and predicting CMV infections in renal transplant recipients, not only qualitative but also quantitative assays must be used together in order to decide the preemptive strategies.