Cloning, expression and characterization of recombinant CagA protein of Helicobacter pylori using monoclonal antibodies: Its potential in diagnostics.

Helicobacter pylori CagA protein plays an important role in the severity of the gastric diseases. Our aims were to clone the cagA 5'- conserved region of the gene, characterize the recombinant CagA (rCagA) protein by monoclonal antibodies (mAbs) and to use this protein for the detection of anti-CagA antibodies by an ELISA test. Our developed rCagA protein (67 kDa) showed an amino acid sequence homology of 83% and 80% with Western and East Asian type strains respectively. Two anti-rCagA (BS-53, CK-02) mAbs and 2 additional rCagA proteins of smaller sizes (60 kDa, 28 kDa) were developed for epitope determination. The BS-53 mAb recognized all 3 rCagA proteins while CK-02 mAb recognized only 2 of them indicating recognition of different epitopes. An in-house indirect ELISA using rCagA was developed to detect anti-CagA antibodies in sera of 59 patients. The ELISA results obtained when compared to those of the PCR gave a sensitivity, specificity and accuracy of 81%, 100% and 88% respectively. We have developed for the first time: a rCagA protein that showed high sequence homology with both Western and East Asian type strains and an indirect ELISA of high performance which can be used to detect anti-CagA antibodies in sera of infected patients worldwide.

Role of γ-glutamyl transferase levels in prediction of high cardiovascular risk among patients with non-alcoholic fatty liver disease.

BACKGROUND & OBJECTIVES: Non-alcoholic fatty liver disease (NAFLD) is an important cause of elevated liver functions. There is evidence showing an association between NAFLD and subclinical atherosclerosis independent of traditional risk factors. We undertook this retrospective study to determine the association of Framingham cardiovascular risk scoring system with liver function tests and inflammatory markers and to find the role of liver function tests in determination of CVD risk among non-obese and non-diabetic subjects with non-alcoholic fatty liver disease. METHODS: A total of 2058 patients were included in the study. Framingham cardiovascular risk scoring was done of all patients according to the age, gender, systolic blood pressure, serum total cholesterol and HDL cholesterol levels, smoking and antihypertensive medication history. Liver function test, lipid profile, insulin, uric acid, ferritin levels, etc. were determined. RESULTS: According to the ultrasonography findings, patients were grouped as without any fatty infiltration of the liver (control group) (n=982), mild (n= 473), moderate (n=363) and severe fatty liver disease (n= 240) groups. In severe fatty liver disease group, the mean Framingham cardiovascular risk score was significantly higher than that of other groups. t0 here was a positive correlation between GGT, uric acid and ferritin levels with Framingham cardiovascular score. In multivariate analysis, high GGT levels were positively associated with high-risk disease presence (OR: 3.02, 95% CI: 2.62-3.42) compared to low GGT levels independent of the age and sex. INTERPRETATION & CONCLUSIONS: Cardiovascular disease risk increases with the presence and stage of fatty liver disease. Our findings showed a positive correlation between elevated GGT levels and Framingham cardiovascular risk scoring system among non-diabetic, non-obese adults which could be important in clinical practice. Though in normal limits, elevated GGT levels among patients with fatty liver disease should be regarded as a sign of increased cardiovascular disease risk. Larger studies are warranted to elucidate the role of GGT in prediction of cardiovascular risk.

Increased microalbuminuria prevalence among patients with nonalcoholic fatty liver disease.

AIM: To determine the prevalence of microalbuminuria, that is an indirect predictor of coronary artery disease, among non-obese and non-diabetic patients with fatty liver disease. MATERIAL AND METHOD: This retrospective study was carried out on non-obese (body mass index (BMI) < 30) and non-diabetic 290 female and 189 male, totally 479 cases. All subjects underwent liver ultrasonography scanning to determine the presence and stage of fatty liver disease. RESULTS: The subjects were grouped according to the ultrasound findings as follows: 182 (37.9%) cases without any fat accumulation in liver were regarded as control group; and among remaining cases, 124 (25.8%) had mild, 93 (19.4%) had moderate, and 80 (16.7%) had severe fatty liver disease. There was not any statistically significant difference between groups in regards to the age, gender, liver function tests, renal function tests or glomerular filtration rate. However urinary protein/creatinine ratio was statistically significantly higher in severe nonalcoholic fatty liver disease (NAFLD) group than the other three groups. In moderate and severe NAFLD groups, microalbuminuria was statistically significantly more common compared with the control and mild NAFLD groups. Regarding the results of multiple logistic regression analysis, presence of fatty liver disease increased the risk of microalbuminuria for 1.87 times independently from increased BMI and increased HOMA-IR values. CONCLUSION: We have determined that microalbuminuria is more prevalent among NAFLD cases compared with control cases and microalbuminuria prevalence was increasing with the advanced stages of NAFLD although two main etiologic factors of microalbuminuria, type 2 diabetes, and obesity were excluded.

MTHFR 677C/T and 1298A/C mutations and non-alcoholic fatty liver disease.

Common genetic mutations encountered in folate metabolism may result in increased homocysteine (Hcy) levels. It has been reported that increased serum Hcy levels may affect the intracellular fat metabolism and may cause enhanced fatty infiltration in the liver resulting in non-alcoholic fatty liver disease (NAFLD). In total, 150 patients diagnosed with FLD by ultrasound examination and 136 healthy control patients that do not have any fatty infiltration in the liver were included in the study. Patients were grouped as mild (n = 88), moderate (n = 38) or severe (n = 24) according to the stage of fatty liver in ultrasound. Serum liver function tests, Hcy, folic acid and vitamin B12 levels of the patients were studied. The genetic MTHFR C677T and A1298C polymorphisms of the patients were also evaluated. Although there was no significant difference in vitamin B12 and folic acid levels, in the severe group, Hcy levels were significantly higher than that of control and mild groups (p<0.001). By contrast, there was no significant difference in heterozygote MTHFR 677C/T and 1298A/C mutations, both MTHFR 677C/T and MTHFR 1298A/C mutations were more common in NAFLD groups compared with the control patients (p<0.001). We have determined increased Hcy levels and increased prevalence of homozygote MTHFR 677C/T and MTHFR 1298A/C mutations in patients with NAFLD compared with healthy controls. Larger studies are warranted to clarify the etiological role of the MTHFR mutations and Hcy levels in FLD.

Influence of obstructive sleep apnea on fatty liver disease: role of chronic intermittent hypoxia.

BACKGROUND: Currently the common pathogenetic mechanisms in nonalcoholic fatty liver disease (NAFLD) and obstructive sleep apnea (OSA) are gaining increased attention. The aim of this study is to find out the influence of chronic intermittent hypoxemia and OSA related parameters to the severity of NAFLD. METHODS: We examined the liver functions tests and ultrasonographic data of liver as well as markers of OSA severity (apnea-hypopnea index [AHI], oxygen desaturation index, minimum oxygen saturation, percentage of time spent with S(pO(2)) < 90%) of 106 subjects. RESULTS: Fatty liver disease was diagnosed in 71 subjects (group 1), and the remaining 35 subjects were taken as controls (group 2). The prevalence of OSA was 71.2% versus 35.7% for group 1 and 2, respectively (P < .001). As NAFLD severity increased from mild to severe form, mean AHI and oxygen desaturation index values also increased significantly. Our multivariate analysis showed that AHI, oxygen desaturation index, lowest desaturation values, and percentage of sleep duration with S(pO(2)) < 90% were independent predictors of NAFLD after adjustment for BMI, weight, and insulin resistance. Furthermore, the most correlated parameter for the severity of NAFLD was found as the duration of hypoxia during sleep. CONCLUSIONS: The prevalence of NAFLD was higher in patients with severe OSA, suggesting a role for nocturnal hypoxemia in the pathogenesis of fatty liver disease.

Patient-controlled analgesia and sedation with alfentanyl versus fentanyl for colonoscopy: a randomized double blind study.

STUDY OBJECTIVE: The aim of this study was to evaluate whether sedo-analgesia with alfentanyl/fentanyl, using a patient-controlled analgesia (PCA) pump, may have positive outcomes in terms of safety, postprocedural workload, and expectations of the colonoscopist, nurse, and patients in elective colonoscopy. PATIENTS: One hundred American Society of Anesthesiology physical status I and II adult patients. INTERVENTIONS: Patients were randomized in a double-blind trial to receive either alfentanyl (n=50) or fentanyl (n=50) by PCA, and incremental doses of midazolam. MEASUREMENTS: Patient expectations were assessed using hemodynamic variables, willingness to have a repeat colonoscopy in the same way, adverse events, discomfort scores, and patient/operator/nurse satisfaction associated with sedo-analgesia. RESULT: All patients in both groups had adequate sedo-analgesia with high satisfaction and willingness scores. There were no serious adverse effects and except for a few events, no required medication. The total sedation times were shorter in the alfentanyl group compared with the fentanyl group. CONCLUSIONS: PCA and sedation with alfentanyl and fentanyl for colonoscopy are safe, feasible, and acceptable to most patients. However, shorter sedation times make alfentanyl more attractive for postprocedural workload.

Helicobacter pylori seroprevalence in patients with chronic prostatitis: a pilot study.

OBJECTIVE: This study investigated the possible relationship between Helicobacter pylori infection and chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). The rate of seropositivity for antibodies against H. pylori was determined in a cohort of subjects with CP/CPPS and prostatitis-free control subjects. MATERIAL AND METHODS: Sixty-four consecutive patients with CP/CPPS and 55 randomly selected asymptomatic men were recruited to the study. Blood samples from enrolled patients and control subjects were analysed using an enzyme-linked Immulite analyser immunoglobulin G serological test for H. pylori diagnosis. Prostate volume, prostate-specific antigen level, maximum urinary flow rate, and International Prostate Symptom Score (IPSS) and National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI) score of the subjects were also determined. The results were analysed with chi-squared and Student's t test and statistical analysis was carried out using SPSS software. RESULTS: There were no significant differences in age and social status between the CP/CPPS and control groups (p > 0.05). Total NIH-CPSI score was significantly higher in the CP/CPPS group. Seropositivity for antibody against H. pylori was higher in the CP/CPPS than the control group (p < 0.05). CONCLUSIONS: This pilot study supports the hypothesis that H. pylori may play a role in CP/CPPS. The infection may be related to the immune response and increased cytokines in seminal plasma and/or expressed prostatic secretion. However, no study has investigated the relationship between CP/CPPS and H. pylori stool antigen positivity. This study showed that H. pylori seropositivity is high in CP/CPPS patients, but this needs to be confirmed by other studies.

Prevalence of proximal neoplasms among asymptomatic patients according to distal colorectal findings.

Colorectal cancer has been described in association with hyperplastic polyposis. Only half of proximal colon cancers are associated with distal adenomas. To compare the prevalence of proximal and advanced neoplasia between patients with distal hyperplastic polyps only; with distal adenomas with or without hyperplastic polyps; and with no distal polyps, we retrospectively analyzed data of 1,064 adults who underwent colonoscopy. Of these patients, 3% had neoplasia. Proximal neoplasia occurred in 0.8% of 945 patients with no distal polyps, compared to none of 19 with distal hyperplastic polyps (P > 0.05) and 6% with distal adenomas (P > 0.05). Proximal advanced neoplasia occurred in 0.6% patients with no distal polyps, compared with none with distal hyperplastic polyps (P > 0.05) and 6% with distal adenomas (P > 0.05). In conclusion, patients with distal hyperplastic polyps, unlike those with distal adenomas, do not exhibit an increased risk for proximal neoplasia or proximal advanced neoplasia compared to those with no distal polyps.

Association of visceral fat accumulation and adiponectin levels with colorectal neoplasia.

PURPOSE: There are few studies addressing the association between measured values of visceral fat accumulation (VFA), adiponectin, and colorectal neoplasia. Our purpose is to investigate the association of VFA and serum adiponectin levels with colorectal adenoma and carcinoma patients. For this purpose, 54 patients with colorectal adenoma and carcinoma, diagnosed by colonoscopic evaluation, and 50 healthy control subjects were included. Patients were subjected to measurement of VFA and adiponectin level and calculation of insulin resistance. RESULTS: Patients with colorectal carcinoma had lower plasma adiponectin levels compared with controls. VFA level did not differ between patients and controls. Adiponectin level was found to be uncorrelated with VFA in the colorectal cancer and adenoma group. No correlation was found between insulin resistance and plasma adiponectin level and VFA. CONCLUSION: Our findings suggest that decreased plasma adiponectin level may be a factor involved in the development of colon cancer or a secondary effect of the metabolic derangements in colorectal cancer.

Eosinophilic colitis in a patient with advanced transitional cell carcinoma of the bladder: a paraneoplastic syndrome?

Eosinophilia associated with tumors is extremely rare. The exact underlying pathogenetic mechanisms are not well understood; limited data have been reported about granulocyte colony-stimulating factor and interleukin-5 in the development of eosinophilia. We report a patient with marked eosinophilia, leukocytosis, and eosinophilic colitis presenting with bloody stool. The subsequent diagnosis was advanced high-grade transitional cell carcinoma of the bladder. Leukocyte count, eosinophil count, and colonoscopic findings returned to normal after surgical removal of the tumor. To our knowledge, eosinophilic colonic infiltration as a manifestation of transitional cell bladder carcinoma has not been previously reported.

Role of Helicobacter pylori in pathogenesis of upper respiratory system diseases.

Helicobacter pylori (H. pylori) is one of the frequently encountered micro-organisms in the aerodigestive tract. Although infections caused by H. pylori are this common, the exact mode of transmission has not been fully understood yet. Oral-oral, fecal-oral and gastrointestinal-oral routes are the possible modes of transmission. This infection is usually acquired in childhood and may persist for the whole life of the patient. However, about 80% of the infected humans are asymptomatic. Human stomach was considered to be the only reservoir of H. pylori until bacteria were discovered in human dental plaque, in oral lesions, in saliva, in tonsil and adenoid tissue. It is suggested that H. pylori enters the nasopharyngeal cavity by gastroesophageal reflux and colonize in the dental plaques, adenoid tissues and tonsils. From these localizations, the bacteria ascend to the middle ear and to the paranasal sinuses directly or by the reflux again and may trigger some diseases, including otitis, sinusitis, phyrangitis, laryngitis and glossitis. But still, the exact mechanism remains unclear.

Hemostasis and global fibrinolytic capacity in chronic liver disease.

Accelerated fibrinolysis associated with liver disease can be demonstrated by various tests that are either nonspecific in liver disease or that demonstrate only an extrinsic pathway. In the present study we used a new method to assess the global fibrinolytic capacity (GFC) of both the intrinsic and extrinsic pathways in patients with chronic liver disease. Forty patients with the diagnosis of chronic liver disease were included in the study. Seventeen age-matched and gender-matched healthy control individuals were enrolled as a control group. The GFC was studied with semiquantitative macrolatex agglutination. The study population consisted of 40 patients with chronic liver disease (group 1, patients with chronic hepatitis; group 2, patients with cirrhosis; group 3, patients with hepatocellular carcinoma), mean age 53.3 +/- 13 years, and a control group (group 4) consisting of 17 healthy individuals (mean age 55 +/- 12.2 years). The GFC was significantly higher in patients than in control individuals (13.8 +/- 9 microg/ml, 13.6 +/- 11 microg/ml, 14.1 +/- 14 microg/ml, 1.9 +/- 2.2 microg/ml, respectively; P < 0.05). There was no difference between the patient groups (P > 0.05). There was a significant positive relationship between the GFC and the prothrombin time and activated partial thromboplastin time values (P < 0.05). A negative correlation was also observed between the GFC and thrombocyte counts (P < 0.05). In conclusion, our results suggest that patients with chronic liver disease have hyperfibrinolysis, as reflected by the increased GFC. Elucidation of the GFC in chronic liver disease can reflect the net fibrinolytic capacity of those patients who are prone to hyperfibrinolysis resulting in bleeding tendencies and hemorrhages.

A prolonged gestational intrahepatic cholestasis: a case report.

Gestational intrahepatic cholestasis, characterized by generalized pruritus and biochemical changes of cholestasis, usually occurs in the third trimester of pregnancy, persists until delivery and resolves spontaneously within the initial four weeks of the puerperium. The incidence is dependent on genetic basis, environmental factors and geographical location. We report the case of a patient with gestational intrahepatic cholestasis with an extraordinary clinical course that extended to the 82nd week postpartum.

Neutrophil-Lymphocyte Ratio (NLR) Could Be Better Predictor than C-reactive Protein (CRP) for Liver Fibrosis in Non-alcoholic Steatohepatitis(NASH).

BACKGROUND-AIM: Non-alcoholic fatty liver disease (NAFLD) is a major cause of chronic liver disease worldwide. The aims of this study were to assess Neutrophil-Lymphocyte Ratio (NLR) and C-reactive protein (CRP), and their association with liver histology in patients with non-alcoholic steatohepatitis (NASH), chronic hepatitis B (HBV), and hepatitis C (HCV). MATERIAL-METHODS: We studied 38 consecutive patients with biopsy-proven NASH, 19 patients with HCV, 45 patients with HBV, and 35 healthy controls who were similar for age and gender. The stage of fibrosis was measured using a 6-point scale. RESULTS: NLR was significantly higher in NASH patients compared to controls, HBV, and HCV patients (p<0.001, p<0.001, and p<0.001, respectively). NLR was positively associated with NAFLD activity scores (r=0.861, p<0.001). NLR was associated with hepatocyte ballooning degeneration (r=0.426, p=0.024), lobular inflammation(r=0.694, p<0.001), steatosis(r=0.498, p=0.007), and fibrosis stage(r=0.892, p<0.001) in NASH patients. Univariate and multivariate analyses showed that NLR was significantly associated with liver fibrosis and NAS (ß=0.631, p<0.001 for liver fibrosis; ß=0.753, p<0.001 for NAS in the multivariate model); however, CRP had no association with liver fibrosis and NAS CONCLUSION: NLR is a promising and inexpensive inflammation marker that correlates with histological grade and fibrosis stage in NASH patients.

Is Helicobacter pylori infection a risk factor for migraine? A case-control study.

BACKGROUND: The relation between Helicobacter pylori and Migraine and usefulness of the eradication treatment on headache is controversial. The aim of this study was to determine whether Helicobacter pylori infection is a risk factor for migraine and whether the eradication of the bacterium can reduce frequency, duration and severity of clinical attacks of the disease. METHODS: A total 70 consecutive patients with migraine with aura and without aura who came to Gastroenterology polyclinic with various complaints were enrolled in the study and compared with a group of 60 matched controls. Helicobacter pylori infection was diagnosed by histopathological biopsies, which was taken by endoscopy (Olympus-GIFXQ240 endoscope). The diagnosis and classification of migraine was made according to the International Headache Society (IHS) criteria. We assessed the frequency, duration and severity of clinical attacks of migraine before and after eradication treatment. The eradication control of Helicobacter pylori was made by Helicobacter pylori fecal antigen test by PCR after 2 months. RESULTS: Helicobacter pylori positiveness is more relevant in the migranous patients compared with controls. 84.6% of patients with eradication treatment and 75% of classically treated patients informed to get benefit from the treatment. CONCLUSION: Helicobacter pylori should be examined in migranous patients and eradication of the infection may be helpful for the treatment of the disease.

Irritable bowel syndrome prevalence in city center of Sivas.

BACKGROUND/AIMS: We planned to determine irritable bowel syndrome prevalence in our region with its distribution according to clinical characteristics of patients and the factors which are considered to be related with irritable bowel syndrome. METHODS: 998 of 1250 individuals (mean age 38.99+0.44) replied to our questionnaire including Rome II criteria. RESULTS: We found the irritable bowel syndrome prevalence in the city center of Sivas to be 19.1%. Distribution of irritable bowel syndrome (+) patients age groups showed no significant difference but irritable bowel syndrome was significantly more common in females. The most common occupation showing irritable bowel syndrome positivity was workers. Irritable bowel syndrome prevalence was also significantly higher in persons not eating three regular meals per day. There was a significantly positive correlation between irritable bowel syndrome prevalence and psychological events, previous abdominal operations and infections. A change in bowel habitus was observed in 41.8% of irritable bowel syndrome patients, and the most common change was constipation. 39.8% of the irritable bowel syndrome patients had applied to the doctor, most often to doctors of internal medicine. We found the irritable bowel syndrome prevalence not to be related with educational status, smoking, daily tea and coffee consumption, alcohol intake, menstrual periods or weight loss. CONCLUSION: Irritable bowel syndrome prevalence in our region with its demographic characteristics was similar to the results seen in western countries.

The relationship between Helicobacter pylori intensity and histopathological findings in cases with chronic gastritis and duodenal ulcer.

BACKGROUND/AIMS: Helicobacter pylori infection affects many people in developing countries. The inflammation it induces can cause malignant gastric lymphoma and also gastric carcinoma, depending on the intensity of inflammatory reaction, specific properties related to the strain and the host response. METHODS: Fifty patients (35 cases with gastritis and 15 with duodenal ulcer) were included in the study. Hematoxylin-eosin paraffin sections were prepared from their endoscopic biopsies and Helicobacter pylori presence, activity of the infection, lymphoplasmacytoid cell infiltration and the degree of atrophy were determined semiquantitatively. Toludin O stain was applied to determine Helicobacter pylori intensity. RESULTS: We found a statistically significant positive correlation of Helicobacter pylori intensity and activity of infection, lymphoplasmacytoid cell infiltration and atrophy in the 35 cases with gastritis and the relationship between Helicobacter pylori intensity and inflammatory activity was statistically significant in the 15 cases with duodenal ulcer. CONCLUSION: As morphological characteristics are crucial for the early diagnosis and treatment of gastric malignities; histopathological confirmation of helicobacter intensity is important.

Alveolar hydatid cyst with lung metastasis: a case report.

Alveolar hydatid disease is a serious and often fatal condition caused by infection with the metacestode form of echinococcus multilocularis. Liver involvement is seen in 90% of the cases. The parasite develops within the liver as a rapid invasive pseudomalignant growth and may make distant metastasis. We present the case of a 37 year old woman admitted to our gastroenterology department with obstructive jaundice and then diagnosed as alveolar hydatid cyst with lung metastasis. This case is presented due to its rare prevalence.

The effect of Helicobacter pylori eradication on symptoms and gastric emptying in patients with nonulcer dyspepsia.

BACKGROUND/AIMS: This study was undertaken to evaluate the effect of eradication therapy on the symptoms of Helicobacter pylori positive non-ulcer dyspepsia patients. METHODS: Twenty-four patients participated in the study and the symptoms of daytime epigastric pain, night or hunger pain, nausea, vomiting, regurgitation, bloating, belching, early satiety and anorexia were scored at the beginning, the 15th day after starting eradication therapy (amoxicillin 2 gr bid, clarithromycin 2 gr bid and omeprazole 40 mg daily for two weeks) and during the third and sixth months. Gastric emptying of radiolabelled solid meal was determined at baseline and during the third month. RESULTS: The Helicobacter pylori eradication rate was 79% and symptom scores significantly decreased during the follow-up period in both of the groups, irrespective of Helicobacter pylori status. The mean symptom scores of the 24 patients at baseline, day 15 and and months three and six were as follows: 1.275, 0.274, 0.496 and 0.238 respectively. Symptom scores for the 19 patients with Helicobacter pylori eradication were 1.084, 0.263, 0.347 and 0.215 respectively while in the five patients in whom Helicobacter pylori eradication therapy failed it was 2.0, 0.314, 1.06 and 0.32 respectively. Of the 16.6% Helicobacter pylori positive non-ulcer dyspepsia patients who had delayed gastric emptying of solids, there was no change after eradication therapy. Nine patients, including all of those in whom eradication therapy failed, required further medication (antacids/prokinetics) for continuing symptoms one month after completion of treatment. CONCLUSIONS: The results of this study suggest that Helicobacter pylori is a causal factor in symptoms of non-ulcer dyspepsia and that eradication therapy improves symptoms and endoscopic findings but has no effect on gastric emptying.