Helicobacter pylori (Hp) is the major recognized risk factor for non-cardia gastric cancer (GC), but only a fraction of infected subjects develop GC, thus GC risk might reflect other genetic/environmental cofactors and/or differences in virulence among infectious Hp strains. Focusing on a high GC risk area of Northern Italy (Cremona, Lombardy) and using archived paraffin-embedded biopsies, we investigated the associations between the Hp vacA and cagA genotype variants and gastric intraepithelial neoplasia (GIN, 33 cases) versus non-neoplastic gastroduodenal lesions (NNGDLs, 37 cases). The glmM gene and the cagA and vacA (s and m) genotypes were determined by polymerase chain reaction (PCR) and sequencing. Hp was confirmed in 37/37 (100%) NNGDLs and detected in 9/33 GINs (27%), consistently with the well-known Hp loss in GC. CagA was detected in 4/9 Hp-positive GINs and in 29/37 NNGDLs. The vacA s1a and m1 subtypes were more common in GINs than in NNGDLs (6/7 vs. 12/34, p=0.014, for s1a; 7/7 vs. 18/34, p=0.020 for m1), with significant vacA s genotype-specific variance. The GIN-associated vacA s1a sequences clustered together, suggesting that aggressive Hp strains from a unique founder contribute to GC in the high-risk area studied.

The insulin/insulin-like growth factor pathway is involved in breast and colorectal cancer (CRC) development. In the present study, we analyzed the coding region and short intron-exon borders of the insulin receptor substrate 1 and 2 (IRS-1 and IRS-2) genes in 12 cell lines derived from breast cancer (BC), 14 cell lines derived from CRC and 33 primary CRCs. The nucleotide variants identified in BC were 3 in IRS-1, 1 of which (p.Arg267Cys) was novel and with a pathogenic potential as predicted by in silico analysis and 6 in IRS-2. Twenty-one variants in IRS-1 and 18 in IRS-2 were identified in the CRC samples. These included 11 novel IRS-1 variants detected exclusively in CRCs, which included 5 missense (p.Pro559Leu, p.Gln655His, p.Asp1014Gly, p.Asp1181His and pPro1203Ser) with a pathogenic potential as predicted by in silico analysis, 2 frameshifts predicted to generate a truncated protein, 1 splice-site mutation and 3 silent variants. In the CRC samples we also identified 7 novel IRS-2 variants, including 4 missense variants, which included 2 (p.Asp782Asn and p.Gly1230Ser) with a pathogenic potential as predicted by in silico analysis, 2 frame insertion mutations and 1 silent variant. Most of the novel IRS-1 and IRS-2 variants may be involved in the modulation of IRS-1 or IRS-2 functions and could be relevant to breast and colorectal tumorigenesis.

To test the hypothesis that Merkel cell polyomavirus (MCPyV) can infect cells of the lymphoid system, we analyzed 353 specimens, including 152 non-Hodgkin lymphomas, 44 Hodgkin lymphomas, 110 benign lymph nodes, 27 lymph nodes with metastasis, and 20 extranodal tissue samples. MCPyV DNA was detected by quantitative PCR in 13 (6.6%) of 196 lymphomas, including 5 (20.8%) of 24 chronic lymphocytic leukemia specimens, and in 11 (10%) of 110 benign lymph nodes, including 8 (13.1%) of 61 samples of reactive hyperplasia and 3 (10.3%) of 29 normal lymph nodes. Other samples were MCPyV negative. Sequence analysis of 9 virus-positive samples confirmed the identity of MCPyV; 3 viral strains were represented. Immunohistochemical testing showed that 1 T-cell lymphoma expressed MCPyV T-antigen. These findings suggest that the lymphoid system plays a role in MCPyV infection and may be a site for MCPyV persistence.

Background: Studies have reported differing frequencies of detection of polyomavirus simian virus 40 (SV40) in association with human lymphomas.Objective: We addressed the hypothesis that SV40 positivity in lymphomas can vary among sampled populations.Study design: Archival paraffin-embedded lymphoma specimens (n = 171) from patients at two urban hospitals in Houston, TX, USA, were analyzed following a cross-sectional study design. Extracted DNAs were characterized by quantitative polymerase chain reaction for the cellular RNase P gene and for SV40 and herpesvirus Epstein-Barr virus (EBV) sequences.Results: Patient characteristics of the two study populations differed significantly whereas the classification of tumor types studied did not. SV40 DNA was detected more frequently in lymphomas from the public hospital population (10/44, 23%) than in lymphomas from the veterans' hospital (VAMC) (4/127, 3%; P<0.0001). EBV detection in lymphomas also differed between the two groups (17/44,39% vs. 23/127, 18%; P=0.01). SV40 positivity was associated with a younger age category of VAMC lymphoma patients (P=0.02). Expression of T-antigen was detected by immunohistochemistry in half of lymphomas that contained SV40 DNA. Variation was observed in the quality and quantity of DNA recovered from paraffin-embedded specimens, but there was no difference in recoveries of DNA from samples from the two hospitals.Conclusions: This study demonstrated that, in a direct comparison, the prevalence of SV40 DNA in lymphomas can differ significantly between groups with different demographic distributions. (C) 2009 Elsevier B.V. All rights reserved.

gastric cancer and duodenal ulcer with homB acting as a factor that could distinguish gastric cancer from duodenal ulcer (adjusted odds ratio: 2.65, 95% confidence intervals: 1.183 ~ 5.934). Conclusions: This study suggests that homB may contribute to development of gastric cancer such that the presence of the homB gene can possibly be used as a factor for discriminating risk of gastric cancer from that of duodenal ulcer in Western countries.

Translocation t(11;18)(q21;q21) is the most frequent chromosomal aberration reported in gastric mucosa-associated lymphoid tissue (MALT) lymphomas. Intriguingly, this translocation has been reported only rarely in diffuse large B-cell lymphomas; it has been proposed that t(11;18)-positive tumors rarely progress to diffuse large B-cell lymphomas. We examined the frequency of chromosomal translocation t(11;18)(q21;q21) in mucosa-associated lymphoid tissue lymphoma and diffuse large B-cell lymphoma of the stomach. Paraffin-embedded tissues from patients with gastric B-cell lymphomas were selected retrospectively. The presence of the t(11;18)(q21;q21) was determined using reverse transcriptase-polymerase chain reaction and/or fluorescence in situ hybridization. beta-Actin transcript was also determined to evaluate the integrity and efficiency of RNA (cDNA) recovery from paraffin-embedded tissues. We analyzed 53 gastric B-cell lymphomas (33 diffuse large B-cell and 20 mucosa-associated lymphoid tissue) obtained from Italy, the USA, or Japan. Beta-actin transcript was amplified in 50 cases (94%), including 19 mucosa-associated lymphoid tissue and 31 diffuse large B-cell lymphomas (five with mucosa-associated lymphoid tissue components). The t(11;18) translocation was detected in 19% (6 of 31) cases with diffuse large B-cell lymphoma versus 26% (five of 19) with mucosa-associated lymphoid tissue lymphoma (P = 0.72). One of five diffuse large B-cell lymphomas with a mucosa-associated lymphoid tissue component showed the t(11;18)(q21;q21). In conclusion, translocation t(11;18)(q21;q21) was found in both mucosa-associated lymphoid tissue lymphomas and diffuse large B-cell lymphomas of the stomach at approximately equivalent frequencies; its presence does not exclude progression to diffuse large B-cell lymphoma.

gastric cancer and duodenal ulcer with homB acting as a factor that could distinguish gastric cancer from duodenal ulcer (adjusted odds ratio: 2.65, 95% confidence intervals: 1.183 ~ 5.934). Conclusions: This study suggests that homB may contribute to development of gastric cancer such that the presence of the homB gene can possibly be used as a factor for discriminating risk of gastric cancer from that of duodenal ulcer in Western countries.

Objective. Chronic infection with Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) has been proposed as a cause of Crohn's disease. Although numerous investigators have examined the link between M. paratuberculosis and Crohn's disease, the evidence remains controversial. The aim of this study was to examine intestinal granuloma from Crohn's patients for M. paratuberculosis using a semi-nested M. paratuberculosis-specific IS900 polymerase chain reaction (PCR). Material and methods. Paraffin-embedded ileal or colonic tissues of patients with Crohn's disease were analyzed. Microdissection of this tissue into “granulomas” and “not granulomas” was performed. On the basis of sequences reported in GenBank alignments, we designed primer sets specific for M. paratuberculosis. The presence of the M. paratuberculosis was examined by semi-nested IS900-specific PCR with human β-actin gene as a control for DNA quality. Results. Biopsies from 20 Crohn's patients were examined. Human β-actin gene was amplified in all samples. M. paratuberculosis DNA was detected in the microdissected granuloma in 1 (5%) patient with Crohn's disease and in none of the “not granuloma” tissues. Conclusions. M. paratuberculosis DNA can rarely be detected within Crohn's granuloma. These results do not support M. paratuberculosis as the primary etiology of Crohn's disease.

Purpose: Crohn's disease is a chronic inflammatory bowel disease of as yet unknown and possibly heterogeneous aetiology. A possible infectious aetiology has also been repeatedly discussed. One of the most plausible candidates is Mycobacterium avium subspecies paratuberculosis (MAP). Numerous authors have explored the link between MAP and Crohn's disease using various methods on biopsies, resections, fresh, frozen, and paraffin-embedded tissue, with variable results. Aim: To examine the detection rate of MAP in patients with Crohn's disease using paraffin-embedded tissues and a semi-nested IS900 specific PCR. Methods: Paraffin-embedded tissues of patients with Crohn's disease were analyzed. Microdissection of granulomas and peri-granulomas (normal) was performed. On the basis of sequence reported in GenBank alignments, we designed primer sets specific for M. avium subsp. paratuberculosis. The presence of the MAP was examined by semi-nested IS900 specific PCR. Results: Human beta-actin gene was amplified in all cases, indicating successful DNA quality. Detection rate of MAP specific IS900 DNA by PCR was appropriately identified from two controls (paraffin-embedded tissue and bacteria culture) confirming the specificity and sensitivity of designed primers. The presence of MAP was found in 1/20 (5%) patients with Crohn's disease; in this case the semi-nested PCR targeting IS900 was positive in microdissected granuloma. Conclusion: We designed a PCR assay that permits the detection of short DNA fragments of the MAP specific IS900 gene in paraffin-embedded samples. Furthermore, the use of microdissection permitted to detect MAP DNA in granulomas in one patient with Crohn's disease. Detection of MAP DNA within granuloma might suggest an infectious etiology in a small subset of patients with Crohn's disease. Acknowledgments: Dr. Sonia Toracchio was supported by a fellowship from Fondazione Italiana per la Ricerca sul Cancro (FIRC), Milan, Italy.

Purpose: Translocation t (11; 18) (q21; q21) is the most frequent chromosomal aberration in extranodal marginal zone lymphoma of the MALT type. This translocation has been associated with an aggressive course but has not been described in high grade gastric MALT lymphoma. Some authors propose that tumors with this translocation rarely or never progress to high grade lymphomas. However, these conclusions are based on examining few specimens. This study examines the frequency of chromosomal translocation t (11; 18) (q21; q21) in lymphomas from patients with low grade and high grade marginal zone lymphoma of the MALT type. Methods: We examined paraffin embedded tissue of patients with gastric marginal lymphoma of MALT type. The presence of the t (11; 18) (q21; q21) was determined using reverse transcription-polymerase chain reaction (RT-PCR); B-actin transcript was amplified to ensure specimen adequacy. Results: We examined 64 paraffin-embedded gastric lymphomas (39 high-grade MALT lymphomas and 22 low-grade MALT lymphomas) from Italy, USA and Japan. The actin transcript was amplified in 61 of 64 cases (95.3%). The t (11; 18) translocation was detected in 21% (8 of 39) cases with high-grade MALT lymphoma vs. 4 of 22 (18%) with low-grade MALT lymphoma [odds 0.861, 95% CI 0.227–3.267]. Conclusions: Translocation t (11; 18) (q21; q21) is characteristic of extra nodal marginal lymphomas of MALT type. It was found in both low-grade and high-grade gastric lymphomas of the MALT type at an approximately same frequency. These results challenges the assumptions that this translocation is absent in high grade lymphoma or that the translocation precludes its progression to high grade lymphoma. Testing for the translocation may provide additional prognostic information or help identify a subgroup where close follow is needed.

An experimental murine model was studied to evaluate the orogastrointestinal colonization of Helicobacter pylori and the animal-to-animal transmission. Balb/C mice were infected with H. pylori and housed with uninoculated mice in cages with and without a grate on the floor. Mice were killed after 7, 14, 30, and 45 days, and samples from the esophagus, stomach, small intestine, colon, and rectum were analyzed for H. pylori by PCR and immunohistochemistry and for histological changes. Bacterial colonization was assessed also by culture from stomach samples. H. pylori was cultured by stomach samples of infected mice at 7, 14, and 30 days. Using PCR and immunohistochemistry, H. pylori was detected in inoculated and uninoculated mice in all areas examined, with an high percentage of positive samples in the esophagus and stomach. Moreover transmission was detected, without differences, regardless of whether mice were housed with or without a grate on the floor, supporting an orooral animal transmission.

Controversial data are available on the duration of action of glyceryl trinitrate after acute and chronic application on anal canal pressure. Our aim was to assess the effect of glyceryl trinitrate at 0.2% and 2% on anal canal pressure before and after eight weeks of treatment. Anal canal pressure was evaluated in 12 patients with chronic anal fissures with an electronic probe with three recording sites before and after the application of glyceryl trinitrate, 120 mg on the external anal verge. Six patients received glyceryl trinitrate at 0.2% and six at 2%. Glyceryl trinitrate 0.2% and 2% equally reduce basal anal canal pressure in all three recording sites (P < 0.001) with major effect on the inner site of the canal toward the rectum, for a 60-min period. Eight weeks after application, the effect of glyceryl trinitrate was unchanged. In conclusion, glyceryl trinitrate ointment at 0.2% and 2%, equally reduces anal canal pressure for 60 min and this effect is kept unchanged after eight weeks of application.