Modified Giemsa Stain Research Articles

Transforming Growth Factor-B1 and Matrix Metalloproteinase-7 Promoter Variants Induce Risk forHelicobacter pylori–Associated Gastric Precancerous Lesions

The expression of growth factors, proteolytic enzymes, fibrogenic factors, and cytokines is altered in the Helicobacter pylori-infected gastric mucosa. Therefore, we aimed to evaluate the association of functional promoter variants of transforming growth factor (TGF)-B1 and matrix metalloproteinase (MMP)-7 genes with gastritis and gastric precancerous lesions. After upper gastrointestinal endoscopy, a total of 130 rapid urease test-positive patients with nonulcer dyspepsia were examined for H. pylori infection using modified Giemsa stain and IgG anti-CagA ELISA. All patients and 200 asymptomatic controls were genotyped for TGF-B1 (-509 C>T) and MMP-7 (-181 A>G) substitutions using PCR-RFLP. The genotype and allele frequencies of TGF-B1 and MMP-7 polymorphisms did not differ between patients and controls (p > 0.05). However, the CagA-positive patients with TGF-B1 -509 T allele had higher risk for gastric atrophy (p = 0.026, odds ratio [OR] = 2.38) and lymphoid follicle development (p = 0.028, OR = 2.29). In addition, CagA-positive patients carrying MMP-7 -181 G allele had risk for lymphoid follicle formation (p = 0.027, OR = 2.30). Thus, the present study revealed significant association of functional MMP-7 and TGF-B1 gene variants toward susceptibility to H. pylori-induced precancerous gastric lesions.

GATA Transcription Factors Regulate the Expression of the Human Eosinophil-derived Neurotoxin (RNase 2) Gene

The transcription factors GATA-1 and GATA-2 have been implicated in promoting differentiation of eosinophilic leukocytes. In this study, we examined the roles of GATA-1 and GATA-2 in activating transcription of the secretory ribonuclease, the eosinophil-derived neurotoxin (EDN/RNase 2). Augmented expression of both GATA-1 and GATA-2 was detected in eosinophil promyelocyte HL-60 clone 15 cells in response to biochemical differentiation with butyric acid. Deletion or mutation of one or both of the two consensus GATA-binding sites in the extended 1000-bp 5' promoter of the EDN gene resulted in profound reduction in reporter gene activity. Antibody-augmented electrophoretic mobility shift and chromatin immunoprecipitation analyses indicate that GATA-1 and GATA-2 proteins bind to both functional GATA consensus sequences in the EDN promoter. Interestingly, RNA silencing of GATA-1 alone had no impact on EDN expression; silencing of GATA-2 resulted in diminished expression of EDN, and also diminished expression of GATA-1 in both butyric acid-induced HL-60 clone 15 cells and in differentiating human eosinophils derived from CD34(+) hematopoietic progenitors. Likewise, overexpression of GATA-2 in uninduced HL-60 clone 15 cells resulted in augmented transcription of both EDN and GATA-1. Taken together, our data suggest that GATA-2 functions directly via interactions with the EDN promoter and also indirectly, via its ability to regulate the expression of GATA-1 in differentiating eosinophils and eosinophil cell lines.

Infectious agents and lymphoma development: molecular and clinical aspects

This review is focused on the role of infectious agents in the development of some lymphoma entities. Associations involving bacterial infections mostly regard marginal zone B-cell lymphomas of mucosa-associated lymphoid tissue (MALT)-type. Some paradigmatic examples of these associations include the Helicobacter pylori-related gastric MALT lymphoma and the more recently reported links between Chlamydophila psittaci and ocular adnexal MALT lymphomas and Borrelia burgdorferi and cutaneous MALT lymphomas. The well-documented association between Epstein-Barr virus infection and related lymphoproliferative disorders are analysed as an example of lymphotropic virus with tumourigenic activity. Molecular, biological and clinical features as well as therapeutic implications of these associations are analysed and future perspectives in this field are discussed.

Recognition of the cattle chromosomes by the Q- and G-banding techniques.

Utilization of the quinacrine mustard fluorescence technique supported by photoelectric recordings, and a modified Giemsa staining technique with trypsin pretreatment has made possible an accurate analysis of the cattle chromosomes. All individual chromosome pairs have been identified, and the most clear-cut distinguishing characteristics have been described.

Indistinguishable cellular changes in gastric mucosa between Helicobacter pylori infected asymptomatic tribal and duodenal ulcer patients.

To investigate the changing pattern of different histological parameters occurring in the stomach tissue of Helicobacter pylori (H pylori) infected tribal populations and duodenal ulcer patients among ethnic Bengalis and correlation of the genotypes of H pylori with different histological parameters. One hundred and twelve adult individuals were enrolled into this study between 2002 and 2004. Among them, 72 had clinical features of duodenal ulcer (DU) from ethnic Bengali population and 40 were asymptomatic ethnic tribals. Endoscopic gastric biopsy samples were processed for histology, genotyping and rapid urease test. Histologically, haematoxylin and eosin staining was applied to assess the pathomorphological changes and a modified Giemsa staining was used for better detection of H pylori. For intestinal metaplasia, special stainings, i.e. Alcian blue periodic acid-Schiff and high iron diamine-Alcian blue staining, were performed. PCR was performed on bacterial DNA to characterize the presence or absence of virulence-associated genes, like cagA, and distribution of different alleles of vacA and iceA. Intraglandular neutrophil infiltration, a hallmark of activity of gastritis, was present in 34 (94%) of tribals (TRs) and 42 (84%) of DU individuals infected with H pylori. Lymphoid follicles and aggregates, which are important landmarks in H pylori infection, were positive amongst 15 (41%) of TRs and 20 (40%) of DU subjects. Atrophic changes were observed in 60% and 27.7%, respectively, among DU cases and tribals (P > 0.003). Metaplastic changes were detected in low numbers in both groups. Moderate to severe density distribution of H pylori in the gastric mucosa was 63% among TRs, whereas it was 62% in DU subjects. There were no significant differences in the distribution of virulence-associated genes like cagA, vacA and iceA of H pylori strains carried by these two populations. Our study showed almost similar distribution of inflammatory cells among asymptomatic tribals and DU Bengali patients. Interestingly, the tribal population are free from any clinical symptoms despite evidence of active histologic gastritis and infection with H pylori strains carrying similar virulence markers as of strains isolated from patients with DU. There was an increased cellular response, especially in terms of neutrophil infiltration, but much lower risk of developing atrophy and metaplastic changes among the tribal population.

Lymphocytic gastritis and celiac disease: evidence of a positive relationship

<h3>Aim</h3> Lymphocytic gastritis (LG) is a histological entity characterised by intense lymphocytic infiltration (≥25 lymphocytes/100 epithelial cells) in the gastric surface and pit epithelium. Its cause has not been established, but an association with <i>Helicobacter pylori</i> infection or celiac disease (CD) has been suggested. The aim of this study was to verify the association of LG in adults with CD, with and without <i>H. pylori</i> infection. <h3>Methods</h3> This prospective study included 63 adults with CD in whom gastric and duodenal biopsy was taken simultaneously. The adults were diagnosed as cases of CD based on the modified ESPGHAN criteria and positive serology. The control consisted of 63 non-celiac adults matched for gender and age (within 2 years) without any gastric or duodenal ulcer in whom upper digestive endoscopy was performed. <i>Helicobacter pylori</i> were recognised in gastric biopsy on H&E sections; a modified Giemsa stain was performed in biopsy suspicious for <i>H. pylori</i>. The results were expressed as number and percentage or mean+SEM. Comparisons of quantitative measurements between groups were performed with Student's t test. <h3>Results</h3> The mean age of CD cases (M:F 27:36) at presentation was 34.7±2.2 years. LG was found in 22 (34.9%) CD cases. CD cases positive for LG had a mean gastric IEL/100 surface epithelial cells of 43.2±3.5, compared with a mean of 10.5±0.7 in negative cases. CD cases not showing LG, however, did show significantly increased gastric IEL compared with non-celiac controls (10.5±0.7 vs 4.9±0.3; <i>p</i><0.0001). Three of 63 (4.8%) CD patients were positive for <i>H. pylori</i>, and none of 22 cases of LG were <i>H. pylori</i> positive. <h3>Conclusions</h3> This study shows support for a pathogenetic relationship between celiac disease and lymphocytic gastritis. Celiac disease without lymphocytic gastritis also showed increased gastric intraepithelial lymphocytes. Histopathologists should be alerted to the distinct appearance of lymphocytic gastritis, and duodenal biopsy may be considered to pick up the celiac disease cases.

Diagnosis and Treatment Guidelines for Helicobacter pylori Infection in Korea

Eleven years has passed since the guideline of the Korean College of Helicobacter and Upper Gastrointestinal Research group for H. pylori infection was produced in 1998. During this period the research for H. pylori has much progressed that H. pylori is now regarded as the major cause of gastric cancer. The seroprevalence of H. pylori in Korea was found to be decreased especially below the age of 40s and in the area of Seoul-Gyeonggi province, and annual reinfection rate of H. pylori has decreased up to 2.94%. In the aspect of diagnostic tests of H. pylori the biopsy is recommended in the body instead of antrum in the subjects with atrophic gastritis and/or intestinal metaplasia for the modified Giemsa staining or Warthin Starry silver staining. The urea breath test is the test of choice to confirm eradication when follow-up endoscopy is not necessary. Definite indication for H. pylori eradication is early gastric cancer in addition to the previous indications of peptic ulcer including scar and Marginal zone B cell lymphoma (MALT type). Treatment is also recommended for the relatives of gastric cancer patient, unexplained iron deficiency anemia, and chronic idiopathic thrombocytopenic purpura. One or two week treatment of proton pump inhibitor (PPI) based triple therapy consisting of one PPI and two antibiotics, clarithromycin and amoxicillin, is recommended as the first line treatment regimen. In the case of treatment failure, one or two weeks of quadruple therapy (PPI+metronidazole+tetracycline+bismuth) is recommended. Herein, Korean College of Helicobacter and Upper Gastrointestinal Research proposes a diagnostic and treatment guideline based on currently available evidence.

Promoter methylation of p16 associated with Helicobacter pylori infection in precancerous gastric lesions: A population‐based study

To investigate the relationship between p16 methylation and Helicobacter pylori infection in precancerous gastric lesions, a population-based study was conducted in Linqu County, a high-risk area of gastric cancer in China. Methylation status of p16 was evaluated by methylation-specific polymerase chain reaction in 920 subjects with precancerous gastric lesions. H. pylori status was determined by 13C-urea breath test and the density of H. pylori in biopsy specimens used for detecting methylation status was assessed by the modified Giemsa stain. The frequency of p16 methylation was significantly higher in subjects with H. pylori positive than those with H. pylori negative in each category of gastric lesion (p<0.001, respectively). Compared with H. pylori negative, the odds ratios (ORs) of p16 methylation were markedly elevated in subjects with H. pylori positive for superficial gastritis (OR, 9.45; 95% confidence interval [CI]: 2.94-30.41), chronic atrophic gastritis (OR, 15.92; 95%CI: 7.60-33.36), intestinal metaplasia (OR, 4.46; 95%CI: 2.44-8.13), indefinite dysplasia (OR, 3.67; 95%CI: 1.90-7.10), and dysplasia (OR, 2.48; 95%CI: 1.02-5.99). Moreover, the frequencies of p16 methylation increased steadily with the severity of H. pylori density in gastric mucosa. Compared with H. pylori negative, the OR of p16 methylation was 1.02-16.13 times higher in subjects with mild H. pylori infection, and 2.69-38.73 times higher in those with moderate/severe infection, respectively. Our findings indicate that p16 methylation was significantly associated with H. pylori infection in precancerous gastric lesions, suggesting that H. pylori infection could potently induce methylation of p16 CpG island.

Lymphocytic Gastritis and Celiac Disease in Indian Children: Evidence of a Positive Relation

Lymphocytic gastritis (LG) is characterized by the presence of > or = 25 lymphocytes/100 epithelial cells in the gastric surface and pit epithelium. An association of LG with Helicobacter pylori infection or celiac disease (CD) has been suggested. The aim of this study was to verify the relation of LG with CD, with and without H pylori infection, in children. A total of 164 children with CD diagnosed between June 2003 and October 2005, in whom gastric and duodenal biopsies were performed simultaneously, were enrolled prospectively. The control group was composed of 164 children without CD, matched for sex and age, who were undergoing upper digestive endoscopy. H pylori was searched for in gastric biopsy specimens sectioned and stained with hematoxylin and eosin, and a modified Giemsa stain for H pylori was performed for confirmation. The Student t test was used to compare quantitative measurements between groups. LG was found in 69 (42.1%) patients with CD. Positive cases had a mean of 43.9 +/- 1.5 intraepithelial lymphocytes per 100 surface epithelial cells, compared with a mean of 13.4 +/- 0.4 in negative cases and 7.8 +/- 0.5 in non-CD control children (P<0.0001). Patients not showing LG did, however, show significantly increased gastric intraepithelial lymphocytes compared with the control children. Nine of 164 CD patients, and 4 of 69 patients with LG, had positive results for H pylori. This study supports a pathogenetic relation between CD and LG. CD without LG also showed increased gastric intraepithelial lymphocytes. H pylori infection may be another cause of LG in children.

Epstein-barr virus in gastric lymphoma- An Indian perspective

AbstractBackground: Primary Gastric Non- Hodgkin′s Lymphomas (NHL) are being increasingly encountered in the recent years. While the role of Helicobacter pylori (H. pylori) has been explored in its causation, some studies have suggested the role of the Epstein - Barr virus (EBV). However data from India is not readily available and this prompted us to study the morphology of primary gastric NHL, the prevalence of EBV infection in primary Gastric NHL and study its association with H. pylori infection. Materials&amp; Methods: A retrospective study. Sixteen biopsy proven cases of primary gastric NHL were studied. Histologic features were reviewed. Immunohistochemistry for EBV LMP-1 and Modified Giemsa stain to identify Helicobacter pylori were done. Results: There were 10 low-grade and 6 highgrade lymphomas, 15 of them classifiable as Mucosa-associated lymphoid tissue (MALT) type, owing to the presence of lymphoepithelial lesions. 36% of the cases were EBV positive. Only one case showed H. pylori. 4 patients succumbed to the disease, of which 3 were seropositive for HIV. Conclusion: Our pilot study suggests that EBV may have an important role in the pathogenesis of gastric MALT lymphomas. The significance of this association and its prognostic value need to be explored in larger number of patients.

Effect of the flavonoid quercetin on inflammation and lipid peroxidation induced by Helicobacter pylori in gastric mucosa of guinea pig

Helicobacter pylori infection induces an inflammatory response in the gastric mucosa. Activation of polymorphonuclear leukocytes can produce oxidative damage to gastric tissue through intermediary radicals of oxygen and nitrogen. Vegetable extracts containing polyphenols of the flavonoid family have antibacterial activity, and the flavonoid quercetin possesses anti-H. pylori activity in vitro. The aim of this study was to analyze the effect of oral administration of pure quercetin on inflammation and lipid peroxidation induced by H. pylori in the gastric mucosa of the guinea pig. Sixty days after oral infection with H. pylori guinea pigs received 200 mg/kg of quercetin daily by mouth for 15 days. The infiltration index of inflammatory cells and bacterial density in both the pyloric antrum and corpus were histologically determined by myeloperoxidase histochemistry, hematoxylin-eosin, and modified Giemsa stains. The lipid hydroperoxide content was assessed by the orange xylenol spectrophotometric method. Quercetin significantly reduced the infiltration index of mononuclear cell and bacterial colonization in the pyloric antrum and corpus. In the antrum of infected quercetin-treated animals, a significant diminution of neutrophil leukocyte infiltration was observed compared with the infected nonquercetin-treated animals. In the antrum, the lipid hydroperoxide concentration was significantly decreased in infected animals treated with quercetin, whereas in the corpus no significant differences were observed. Our results indicate that in vivo oral quercetin administration decreases H. pylori infection in the gastric mucosa and reduces both the inflammatory response and lipid peroxidation.

Endoscopic Findings And The Frequency Of Helicobacter Pylori Among Dyspeptic Patients In Maiduguri, North-Eastern Nigeria

Aim: To determine the types of upper gastrointestinal lesions and the frequency of Helicobacter pylori infection among dyspeptic patients in Maiduguri. Methods: Three hundred and thirty (330) dyspeptic patients undergoing endoscopy at the University of Maiduguri Teaching Hospital were prospectively studied. Gastric antral biopsy specimens obtained through endoscopic biopsy forceps were stained with haematoxylin and eosin. Modified Giemsa staining was employed to demonstrate the presence Helicobacter pylori. Results: One hundred and seventy one (51.8%) of the patients were males and 159 (48.2%) were females. Their ages ranged from 14 to 80 years with a mean of 38.7, the age group with highest frequency being 40 to 49 years. A total of 259 (78.5%) of the patients were positive for H. pylori. Two hundred and seventy one (82.1%) had endoscopically identifiable lesions. The most common lesion was oesophagitis which was seen in 67.0% of the patients followed by gastritis (60.0%) and duodenitis (27.9%). Gastric ulcer was recorded in 6.4 % of the patients while 4.8% and 1.7% had duodenal ulcer and gastric cancer respectively. The frequency of H. pylori among patients with gastritis was 89.1% while it was 83.3%, 100% and 61.9 % among patients with duodenitis, duodenal ulcer and gastric ulcer respectively, and 33.3% among those with gastric cancer. Conclusion: There is high frequency of H. pylori infection among dyspeptic patients in Maiduguri. The predominant endoscopic findings were oesophagitis, gastritis and duodenitis. Keywords: Helicobacter pylori, dyspepsia, endoscopy, frequencyHighland Medical Research Journal Vol. 5 (1) 2007 pp. 78-81

Genetic association of interleukin-1 haplotypes with gastritis and precancerous lesions in North Indians

We evaluated the association of functional variants of IL-1 genes with the development of gastritis and precancerous lesions, which are known to be influenced by inflammatory response against Helicobacter pylori. After upper gastrointestinal (GI) endoscopy, 120 patients with gastritis were tested for H. pylori infection using rapid urease test, modified Giemsa staining and IgG anti-CagA ELISA. All patients and 243 healthy controls were genotyped for IL-1B (-511 C/T) and IL-IRN (VNTR) genes using PCR-RFLP/PCR. IL-1B: (-511 C/T) genotype/allele were not associated with gastritis. IL-1RN 1/2 genotype carriers had susceptibility to gastritis (p=0.025, OR=1.7). Individuals with the IL-1RN 1/1 genotype (p=0.05, OR=0.65) and IL-1B -511*T-IL-1RN *1 haplotype were at low risk for gastritis (p=0.043, OR=0.72). High secretor haplotype combinations (C1-/T2+, C1-T1+ and T1+/T2+) did not influence neutrophilic infiltration, glandular atrophy or intestinal metaplasia. We identified that individuals with the IL-1RN 1/2 genotype had increased risk for gastritis. IL-1B -511*T-IL-1RN *1 (T1) haplotype carriers were at decreased risk for gastritis and no significant association was observed for precancerous lesions in North Indians.

Risk of lymphoid follicle development in patients with chronic antral gastritis: role of endoscopic features, histopathological parameters, CagA status and interleukin-1 gene polymorphisms

Helicobacter pylori infection causes gastritis, lymphoid follicle formation and development of MALT lymphoma. We evaluated endoscopic, histological, serological and genetic risk factors associated with lymphoid follicle development in gastritis. After upper GI endoscopy, 3 antral biopsies were taken from 120 patients for histological examination. H. pylori was diagnosed using rapid urease test (RUT), modified Giemsa stain and IgG anti-CagA ELISA. Genotyping of IL-1B (-511C/T) and IL-1RN (86 bp VNTR) genes were performed by PCR-RFLP/PCR. In 120 patients, 45 (37.5%) showed presence of lymphoid follicles in antral gastric mucosa. H. pylori was positive by modified Giemsa stain (26%) RUT (50%) and anti-CagA IgG in 67.5%, The presence of nodularity (p = 0.030), neutrophilic infiltration (p = 0.010), lymphocytic infiltration (p = 0.002), glandular atrophy (p = 0.0001), glandular shortening (p = 0.001), fibrosis (p = 0.0001), plasma cells (p = 0.007), eosinophils (p = 0.012), anti-CagA antibodies (p = 0.003) and H. pylori density (p = 0.020) were associated with risk (odds ratio = 11.5, 3.8, 11.0, 8.4, 3.8, 4.6, 5.8, 16.0, 10.8 and 2.8 respectively) of lymphoid follicle. However, IL-1 gene polymorphisms did not influence lymphoid follicle development The presence of modularity, lymphocytic infiltration, glandular atrophy, glandular shortening, fibrosis, plasma cells, eosinophils and anti-CagA IgG antibodies are risk factors for lymphoid follicle development in patients with gastritis.

Association of Toll-like receptor–4 (Asp299Gly and Thr399Ileu) gene polymorphisms with gastritis and precancerous lesions

A Toll-like receptor-4 (TLR-4) Asp299Gly and Thr399Ileu substitution reduces responsiveness to Helicobacter pylori (H. pylori) lipopolysaccharide. CagA+ strains of H. pylori are known to be associated with gastroduodenal diseases. Therefore we aimed to evaluate association of TLR-4 substitutions and CagA seropositivity with gastritis and precancerous lesions in a northern Indian population. After upper gastrointestinal endoscopy, 130 rapid urease test (RUT)-positive patients with nonulcer dyspepsia (NUD) were included. Patients with NUD were also screened for H. pylori infection using modified Giemsa staining and anti-CagA IgG enzyme-linked immunoabsorbent assay. All patients and 200 asymptomatic control subjects were genotyped for TLR-4 substitutions using polymerase chain reaction-restriction fragment length polymorphism. We observed that frequencies of TLR-4 Asp299Gly variants were comparable between patients and control subjects, and also between positive and negative groups of precancerous lesions in patients. Frequencies of TLR-4 399Ileu allele (8% vs 3%, p = 0.008) and Asp299-Ileu399 haplotype (6.5% vs 3%, p = 0.022) were higher in patients than in control subjects at risk for gastritis (OR = 2.6 and 2.5, respectively). TLR-4 399Ileu allele carriers had higher risk for plasma cell infiltration (p = 0.023, OR = 10.6) that led to atrophy (p = 0.028, OR = 4.2) and intestinal metaplasia (p = 0.009, OR = 4.7). CagA positivity was more frequently associated with lymphoid follicle formation (p = 0.033, OR = 2.53). In conclusion TLR-4 Thr399Ileu substitution may be a risk factor for gastritis and precancerous lesions. CagA positivity may be a risk factor for lymphoid follicle development but not for other precancerous lesions in a northern Indian population.

Interleukin-10 (−819 C/T) and Tumor Necrosis Factor-α (−308 G/A) Gene Variants Influence Gastritis and Lymphoid Follicle Development

Helicobacter pylori (H. pylori) causes gastritis, development of lymphoid follicles and later monoclonal mucosa-associated lymphoid tissue (MALT) lymphoma. We evaluated the association of tumor necrosis factor (TNF)-alpha (-308 G/A) and IL-10 (-819 C/T) gene polymorphisms with gastritis and lymphoid follicle formation. H. pylori infection was detected using modified Giemsa staining and IgG anti-CagA enzyme-linked immunosorbent assay (ELISA). One hundred and thirty patients with non-ulcer dyspepsia (NUD) and 200 healthy age-matched controls were genotyped for TNF-alpha and IL-10 polymorphisms using polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP). Subjects with IL-10 -819 T allele [patients (46.5%) versus controls (35.7%), p = 0.006, OR = 1.56, 95% CI = 1.14-2.15] were at risk of gastritis. Infection with H. pylori was more often associated with lymphoid follicles formation than its absence (46% versus 22%, p = 0.009). TNF-alpha polymorphism did not influence gastritis but patients with TNF-alpha -308 A allele carriers showed >2 fold risk of lymphoid follicle formation [presence (26%) versus absence (11.25%), p = 0.029, OR = 2.8; 95% CI = 1.09-7.08]. There was a trend towards association of lymphoid follicles and TNF-alpha -308 A allele carriers with H. pylori infection than without (58.5% versus 22.2%; p = 0.064). IL-10 -819 T and TNF-alpha -308 A alleles may increase risk of gastritis and lymphoid follicle formation.

The Hungarian epidemiology of clarithromycin resistance in Helicobacter pylori infection

Introduction: Clarithromycin is a main component of the eradication regimens for Helicobacter pylori infection. Aims: The purpose of the study is the assessment of the current prevalence of clarithromycin resistance in our district and an overview of the Hungarian data. Methods: A). From the patients examined at the Department of Gastroenterology of Ferencváros Health Center, biopsy samples were randomly selected from 238 cases. The presence of Helicobacter pylori was confirmed by the modified Giemsa stain and rapid urease test. The resistance to chlarithromycin was determined by fluorescence in situ-hybridization. B) Hungarian articles and congress abstracts dealing with resistance to clarithromycin, published between 1995–2006 were reviewed and the data concerning the prevalence of resistance were extracted and analysed. C) The data concerning clarithromycin use since its introduction in 1993 were obtained from the National Institute of Pharmacology. Results: A) The prevalence of primary clarithromycin resistance was 17.3%. The resistance was complete in 47.4% and partial in 52.6%. Secondary resistance in patients with known previous treatment with macrolides was 55.5%. There was a weak positive correlation between age (r=0.15), female gender (r=0.10) and smoking (r=0.16) and prevalence of resistance, while alcohol consumption, the diagnosis and histological severity of the infection did not influence it. B) Claritromycin resistance was addressed in 8 papers, including a total of 775 cases. The prevalence of resistance determined by phenotypic methods was 3.9%, while fluorescence in situ-hybridization detected the resistance in 17.0% of the cases. Regional differences were also encountered. Secondary resistance was met in 55.5% using phenotypic and in 49.0% with genotypic methods. C) The use of clarithromycin increased fivefold between 1993–2005. Discussion: The prevalence of clarithromycin resistance determined by genotypic method is increased as compared to the results of earlier phenotypic methods, which could be due to the more extensive use of macrolides.

Role of Endoscopic Ultrasound in the Diagnosis of Intraductal Papillary Mucinous Neoplasms: Correlation With Surgical Histopathology

Intraductal papillary mucinous neoplasms (IPMNs) are precancerous tumors characterized by dilation of the main pancreatic duct, its side branches, or both. The purpose of this study was to evaluate the role of endoscopic ultrasound (EUS) in differentiating benign and malignant IPMNs. We identified all patients between July 1996-November 2005 who underwent preoperative EUS for IPMNs. Malignancy was defined as the presence of invasive carcinoma; all other neoplasms were considered benign. The results of EUS and EUS-guided fine-needle aspiration (EUS-FNA) were compared with corresponding histopathology. Seventy-four patients (38 male; mean age, 65 years) with 21 (28%) malignant and 53 (72%) benign IPMNs were identified. Sixty-five (88%) underwent EUS-FNA. Compared with benign tumors, patients with malignant IPMNs were more likely to be older (P = .011), present with jaundice (P = .03) or weight loss (P = .03), and have EUS features of a dilated main pancreatic duct (P = .0001), solid lesion (P = .0001), pancreatic ductal filling defects (P = .03), or thickened septa within any cyst (P = .02). The sensitivity, specificity, and accuracy of EUS-FNA for the diagnosis of malignancy were 75% (95% confidence interval [CI], 53%-89%), 91% (95% CI, 79%-97%), and 86% (95% CI, 76%-93%), respectively. Cyst or pancreatic duct fluid carcinoembryonic antigen (CEA) and carbohydrate antigen (CA) 19-9 did not differ between groups. Older age, jaundice and weight loss, and EUS features of a solid lesion, dilated main pancreatic duct, ductal filling defects, and thickened septa are predictive of malignancy in patients with IPMNs. EUS-FNA cytology is helpful, but cyst fluid CEA and CA 19-9 are of limited value to differentiate malignant from benign IPMNs.

Endoscopic Ultrasound–Guided Fine-Needle Aspiration of Ascites

The aim of this study is to report a large single-center experience with endoscopic ultrasound (EUS)-guided fine-needle aspiration (FNA) of ascites. Consecutive patients at our institution in whom EUS-guided paracentesis was performed between January 1997 and July 2005 were identified retrospectively. All procedures were performed by or under the supervision of 1 of 5 experienced endosonographers with available on-site cytopathology. Sixty consecutive patients (33 men; mean age, 67 y) were identified. Previously attempted percutaneous paracentesis was unsuccessful in 3 of 6 patients. Ascites confirmed by EUS FNA was visible in 28 of 54 (52%) computerized tomography, 3 of 11 (27%) transabdominal ultrasound, and 4 of 8 (50%) magnetic resonance imaging examinations before EUS. Transgastric (n = 55) or transduodenal (n = 5) EUS-guided paracentesis (mean, 8.9; range, 1-40 mL) revealed malignancy in 16 (27%) from primary pancreatic (n = 9), gastric (n = 2), urothelial (n = 1), esophageal (n = 1), gallbladder (n = 1), bile duct (n = 1) cancer, and lymphoma (n = 1). The cytology from 2 patients was atypical (1 suspicious for malignancy and 1 considered reactive) and the remaining 42 were benign. Potential complications occurred in 2 of 60 (3%) patients with self-limited fever. Of the 8 of 60 (13%) patients who underwent subsequent surgery, 3 had metastatic pancreatic adenocarcinoma (n = 2) and metastatic small intestinal carcinoid (n = 1) to the peritoneum after negative EUS-FNA cytology. EUS frequently identifies ascites missed by other imaging studies. EUS-guided paracentesis may identify malignancy in a subset of patients. Negative ascitic fluid cytology from EUS FNA does not exclude possible peritoneal carcinomatosis.

Seroprevalence of Microsporidiosis in Immunocompromised Patients in Kano-Nigeria

Microsporidial infections have been recognized as an increasingly important infection in immuncompromised patients, particularly those infected with HIV/AIDS. Stool samples were examined for microsporidial spores by modified Giemsa staining technique and IgG antibodies to Microsporidia in Sera samples were detected by ELISA. Feacal examination showed that 14/192 (7.29%) of HIV/AIDS patients had microsporidia, comprising Enterocytozoon bieneusi 8/14 (57.14%), Encephalitozoon intestinalis 5/14(35.71%) and mixed infection of both 1/14(7.14%), while 0/50 of HIV-negative patients had microsporidia. The difference between them was not significant (X, p>0.05). Enterocytozoon bieneusi was detected in 3/23(13.04%) and 3/35(8.57%) of TB/HIV/AIDS and TB/HIV-negative patients. Serological evaluation showed that 22/168(13.10%) of HIV/AIDS and 2/60 (3.33%) of HIV-negative patients were seropositive. There was a significant difference (X, p<0.05) in their infection rates. 0/30 of TB patients were seropostive. There was a significant association (p<0.05) between patients with Microsporidia and CD4 lymphocytes counts of ≤ 50 cells/μl and also with diarrhoea. Detection of Microsporidia in Immunocompromised patients has not been described previously in this area.