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Biomedical subjects

M Rugge

Publications and source records attributed to M Rugge.

At least 91 records · Page 5Linked to original sources

Gastric juice polymerase chain reaction: an alternative to histology in the diagnosis of Helicobacter pylori infection.

BACKGROUND: Infection from Helicobacter pylori plays a role in several gastroduodenal diseases. The recent availability of molecular techniques, particularly the polymerase chain reaction (PCR), allows us to detect small amounts of this bacterium. The aims of this study were to compare PCR and histological findings and to ascertain the clinical usefulness of H. pylori PCR identification in different biological samples. MATERIALS AND METHODS: We studied 94 consecutive patients. Saliva, gastric juice, and four antral and four body biopsies were obtained from each patient. H. pylori was evaluated histologically in two antral and two body biopsies (Giemsa or Warthin-Starry stain). After extraction, DNA was submitted for PCR amplification using the two primers HPU1 and HPU2, which amplified a 411-bp product from the urease gene A. RESULTS: Forty-nine patients were H. pylori-positive at histological workup. The sensitivity of PCR was 92% for gastric juice, 73% for antral biopsies, 61% for body biopsies, and 13% for saliva. Of the 45 H. pylori-negative patients at histological assessment, 7 (16%) had positive findings on PCR, mainly when gastric juice was examined. CONCLUSIONS: These results indicate that PCR is as sensitive as histological assessment. We suggest that PCR H. pylori detection in gastric juice is a sensitive method for diagnosing this infection.

Adult↗

Hepatitis C virus serotypes and liver pathology.

The present study aimed to analyze the pathology of chronic hepatitis C in relation to HCV serotype, and to speculate on possible differences in the pathogenesis of liver injury. Liver biopsies were investigated from 59 consecutive patients in whom hepatitis C virus genotypes were determined by a serological genotyping assay that detects antibodies directed to epitopes encoded by the NS4 region. A morphological study was performed in each case, semiquantitatively scoring necro-inflammatory and fibrotic liver lesions. The prevalence of HCV serotypes was as follows: 26 of the 59 patients (44%) had type 1 infection, 11 (19%) had type 2 and 20 (35%) had type 3. A significant association between intravenous drug abuse and serotype 3 infection was observed. Patients with HCV type 2 proved significantly older than patients with infection type 1 or 3, and more frequently they showed a more active liver disease, but no differences were found in the quality and acinar topographic distribution of all the morphological lesions scored. In conclusion, in chronic hepatitis C a more active liver disease can be related to HCV serotype 2 but the spectrum of liver lesions is independent of HCV types. From a morphological point of view, a different pathogenesis of liver injury related to different HCV types is unlikely.

Adult↗

Is there a need for gastrointestinal pathologists? Gruppo Italiano di Patologi dell'Apparato Digerente.

The interdisciplinary approach to medical practice is a cultural and operational prerequisite for proper diagnostic and therapeutic activity. Its constant application contributes to the improvement of medical performances with beneficial effects on their costs. Anatomic pathology interacts with clinical and instrumental diagnostics and makes a fundamental contribution to the conclusive diagnosis of disease. Furthermore, morphological data are often crucial in the monitoring of therapeutic effects and a mandatory prerequisite for evaluating the efficacy of treatment in clinical trials both on neoplastic and non-neoplastic diseases. Such deep involvement in clinical practice and the need to keep diagnostic practice up to date with the explosive advances in scientific knowledge justify the demand for pathologists specializing in sub-specialties of their discipline. However, there is an inevitable need for these dedicated pathologists to remain within the cultural and physical environment of the Pathology Department where they have been introduced to and trained in the discipline. The present paper highlights the scientific and practical motives leading to the creation in Italy, and other European countries of a working group open to pathologists particularly dedicated to gastrointestinal disease. The group is called the Gruppo Italiano di Patologi dell'Apparato Digerente (GIPAD).--Italian Group of Digestive Tract Pathologists.

Digestive System↗

Azithromycin for the cure of Helicobacter pylori infection.

OBJECTIVES: Azithromycin, a new antibiotic chemically related to erythromycin, has been proposed for the cure of Helicobacter pylori, achieving high gastric tissue levels (above the MIC for H. pylori) after oral administration. The aim of the study was to establish whether azithromycin plus metronidazole in association with either omeprazole or bismuth subcitrate is useful in curing H. pylori infection of the stomach. PATIENTS AND METHODS: The study involved 132 dispeptic patients who proved to be H. pylori infected by antral and corpus histology (Giemsa, modified) and rapid urease test (CLOtest); the Sydney system was used to classify the gastritis. Sixty-three patients received bismuth subcitrate 120 mg q.i.d. for 14 days plus azithromycin 500 mg o.d. for the first 3 days plus metronidazole 250 mg q.i.d. for the first 7 days; 69 patients received omeprazole 40 mg for 14 days plus azithromycin 500 mg o.d. for the first 3 days plus metronidazole 250 mg q.i.d. for the first 7 days. Patients were well matched for common clinical variables. Cure of H. pylori infection was assessed by the same methods 2 months after completion of treatment. RESULTS: Eleven patients dropped out of the study, only one reporting side effects (nausea, vomiting, and epigastric pain). Cumulative "per protocol" cure rate was 66.1% (CI 95%, 58.5-75.3%). There was no statistically significant difference between the two treatment groups: 58.9% (CI 95% 48.4-74.6%) versus 72.3% (CI 95%, 60.7-82.5%). Intention to treat does not substantially modify results. Few side effects were recorded. Cured patients showed a significant reduction in the activity of gastritis. CONCLUSION: Azithromycin, combined with omeprazole and metronidazole, the cure rate of H. pylori was about 70%. The cure of H. pylori infection improves the activity of gastritis.

Adult↗

Helicobacter pylori serology in patients with chronic gastritis.

OBJECTIVES: Helicobacter pylori (Hp) infection is known to cause several gastroduodenal diseases. In patients with non-ulcer dyspepsia, we assessed the link between Hp infection and gastric mucosal inflammation, as well as the influence of Hp and inflammation on the serum levels of anti-Hp antibodies (IgG), pepsinogen A (PGA), pepsinogen C (PGC), and gastrin. METHODS: Entering the study were 221 patients with non-ulcer dyspepsia, all of whom underwent upper gastrointestinal endoscopy. RESULTS: Of the 221 patients investigated, 135 (61%) were Hp positive. The higher the bacterial load, the worse the associated gastritis, the gastric antrum and body being considered. All of the serological indices studied were found to be influenced by gastritis. Serum IgG satisfactorily discriminated between Hp-positive and Hp-negative subjects, with a sensitivity of 84% and a specificity of 86%. PGC, PGA, and gastrin were less accurate. Only PGC only found to be correlated with Hp load. The product of IgG and PGC improved the diagnostic accuracy of IgG alone. CONCLUSIONS: Hp infection, frequently found in patients with non-ulcer dyspepsia, is associated with gastric mucosal inflammation; of the indices studied, serum IgG and PGC most accurately indicated Hp infection, and their product may be proposed as an aid in diagnosing Hp infection in dyspeptic patients.

Adult↗

[The role of the laboratory in the diagnosis and monitoring of Helicobacter pylori infection].

The diagnosis of Helicobacter pylori infection represents a fundamental step for a correct clinical approach to the patient with gastritis, peptic ulcer or gastric adenocarcinoma or lymphoma. Upper gastrointestinal endoscopy has first to be made in order to obtain gastric juice and mucosal biopsies, where Hp can be found. Two different procedures are recommended to diagnose Hp infection (e.g. urease test and histology or culture). The identification of specific DNA sequences with the polymerase chain reaction represents a sensitive and specific method to diagnose Hp infection. Serum anti Hp antibodies, pepsinogen A and C determination is recommended at diagnosis and during the follow-up to assess the success of therapy.

Chronic Disease↗

Cysteine and serine proteases in gastric cancer.

BACKGROUND: Cysteine proteases (cathepsin B [CATB] and cathepsin L [CATL]), the serine protease urokinase-type plasminogen activator (UPA), and plasminogen activator inhibitor type-1 (PAI-1) are thought to play an important part in cancer invasion and metastasis. The aims of this study were to measure CATB, CATL, UPA, and PAI-1 in gastric cancer (GC) and normal mucosa distant from the tumor (NORM); to evaluate whether tissue levels are related to tumor stage, grade, or histotype; to assess their prognostic relevance; and to examine UPA and PAI-1 expression immunohistochemically. METHODS: Gastric cancer and NORM samples were obtained from 25 patients with gastric cancer patients undergoing surgery (17 males, 8 females; mean age, 62 years; range, 31-84 years). Antigen concentrations were measured using the enzyme-linked immunosorbent assay method. Immunohistochemistry was performed using monoclonal UPA and PAI-1 antibodies. RESULTS: Significantly higher antigen levels were found: (1) in GC vs. NORM (CATB, CATL, UPA, PAI-1) tissues; (2) in GC with versus without metastasis (CATB, CATL, UPA); (3) in poorly or moderately versus well differentiated GC; and (4) in diffuse versus intestinal-type GC (CATB, CATL). Urokinase-type plasminogen activator, PAI-1 and CATB levels had a significant prognostic impact. Cancer and stromal cells, showed immunoreactivity to anti-UPA and anti-PAI-1 antibodies. CONCLUSIONS: These results confirm the important role of CATB, CATL, UPA and PAI-1 in gastric cancer progression. Higher levels are detected in GC with metastases, poorer differentiation, and diffuse histotype, thus identifying patients with a worse prognosis.

Adenocarcinoma↗

Gastric epithelial dysplasia. How clinicopathologic background relates to management.

BACKGROUND: Gastric epithelial dysplasia (GED) in metaplastic mucosa is considered the most advanced preinvasive lesion in the multistep morphogenesis of intestinal-type gastric cancer (GC). The rate of GED's evolution into GC is still under debate and probably is related to pathologic and clinical parameters other than the dysplasia itself. The aim of this study was to evaluate whether clinical aspects (sex and age) and/or morphologic variables (GED grade, coexisting atrophic gastritis) are relevant to the outcome of dysplasia, with a view toward initiating the establishment of a rational follow-up protocol for practical GED management. METHODS: Ninety-three patients harboring GED (G1: 56, G2:34, G3:18) were followed for more than 12 months according to a previously-agreed protocol. Regression, progression, or evolution into GC were detected for each grade of GED. Multivariate analysis was used to check the independence of clinical and pathologic variables in the progression of GED into more severe dysplastic lesions and/or as risk factors for evolution into GC. RESULTS: Age, male sex, GED grade and grade of coexisting atrophic gastritis proved independent risk factors for GED progression, with no significant interactions. Only GED grade (G2 and G3) was significantly associated with carcinomatous evolution. In G1-GED, age and the grade of coexisting atrophy proved to be independent risk factors for carcinomatous evolution. CONCLUSIONS: In G1-GED, more stringent follow-up should be recommended for older patients with coexisting high grade atrophic gastritis; stringent follow-up is always mandatory for G2-GED; and a surgical approach is justified in G3-GED.

Adenocarcinoma↗

[Role and behavior of cathepsin B and cathepsin L in gastric cancer].

Cathepsin B and cathepsin L--cysteine proteinases--may play an important role in cancer invasion and metastasis. The authors determined tissue antigen concentrations of cathepsins, using the ELISA method, in 25 patients with gastric cancer (17 males, 8 females, mean age 62, range 31-84). They evaluated the possible relationship that these proteinases may have with the presence of metastases, differentiation and histotype. Significantly higher cathepsin B and cathepsin L antigen levels were found: 1. in gastric cancer tissues vs. normal tissues distant from tumors (CATB: p < 0.05, CATL: p < 0.005); 2. in diffuse vs. intestinal type cancers (p < 0.05); 3. in patients with poorly vs. well-differentiated cancers (p < 0.05); in gastric cancers with vs. without metastasis (p < 0.05). Their results confirm that cathepsin B and L play an important role in gastric cancer invasion and metastasis. Considering the significantly higher cathepsins detected in cancers with metastasis, a poor differentiation and of diffuse histotype, these proteinases could be useful for identification gastric cancer patients with a poor prognosis.

Adult↗

TP53 gene mutations in gastric carcinoma detected by polymerase chain reaction/single-strand conformation polymorphism analysis of archival material.

TP53 gene mutations, one of the most common alterations described in human tumors, have also been detected in gastric carcinoma, and shown to occur rather late in disease progression. A better assessment of the prognostic value of TP53 gene mutations can be obtained by examining archival material, as this allows stored cases with well-defined histories to be monitored. We performed immunohistochemical and polymerase chain reaction/single-strand conformation polymorphism (PCR-SSCP) analyses of formalin-fixed paraffin-embedded material from nine selected cases of gastric carcinoma at different pathological stages. PCR-SSCP analysis of TP53 exons 5-8 detected missense point mutations in two out of five immunostain(PAb1801)-positive tumors, and a deletion (allowing for a premature stop codon) in one of the remaining four immunostain-negative tumors. Thus, PCR-SSCP analysis represents a feasible strategy for the detection of TP53 alterations in archival material of gastric carcinoma cases.

Aged↗

Influence of Helicobacter pylori on tryptase and cathepsin D in peptic ulcer.

We here ascertain whether tryptase (a serine endoprotease released by mast cells) and cathepsin D (CD, a lysosomal hydrolase that seems able to derange the extracellular matrix) play a part in peptic ulcer disease and whether they are linked to Helicobacter pylori (Hp) infection. We studied 13 controls, 25 patients with gastric ulcer, 47 with duodenal ulcer, and 11 with duodenitis. Tryptase and CD were measured in mucosal biopsies (body and antrum of the stomach and duodenum) using IRMA methods. Hp infection was histologically evaluated (Giemsa). Tryptase and CD levels were higher (25%) in patients with active peptic ulcer, whether gastric or duodenal. In Hp-positive patients the CD mucosal content was higher while tryptase mucosal levels were lower than in Hp-negative patients. Tryptase was correlated with gastrin content. CD seems to be mainly related to the phlogistic reaction of the mucosa to Hp infection; tryptase may reflect an indirect link between Hp infection, gastrin release, and the function of mast cells.

Adult↗

Gastrin stimulates gastric mast cells in rabbits.

In a previous study we demonstrated that in human gastric mucosa tryptase was localized only in mast cells and that its levels were correlated with serum gastrin, suggesting a link between gastrin action and mucosal mast cell function. The aim of the present study was to discover whether pentagastrin injection could stimulate gastric mucosal mast cells in rabbits. Ten female rabbits (group S) were injected s.c. with pentagastrin (10 mu g/kg); another group of ten animals (group C) was injected s.c. with an equal volume of saline solution. One hour after the injection the rabbits were sacrificed and their stomachs removed. Antrum (A), corpus (C) and fundus (F) mucosal homogenates were assayed for total protein, tryptase, pepsinogen A (PGA), histamine and gastrin. Histamine tissue levels were significantly lower in group S than in group C in the antrum (Mann-Whitney test: U = 82, P < 0.01) and in the corpus (U = 83, P < 0.005). Tryptase levels were significantly higher in group S than in group C in all gastric areas (antrum: U = 95, P < 0.001; corpus: U = 85, P < 0.005 and fundus: U = 75, P < 0.05). Total protein, PGA and gastrin did not vary significantly between groups. In group C, no significant correlations were found among the five parameters. In group S, corpus tryptase was correlated with fundus tryptase (Spearman's r = 0.831, P < 0.01). The same relationship was observed for histamine (r = 0.672, P < 0.05). In group S, antrum gastrin was inversely correlated with antrum tryptase (r = -0.903, P < 0.001), and with corpus PGA (r = -0.806, P < 0.05). This study demonstrates that bolus pentagastrin administration stimulates gastric mucosal mast cells in the rabbit.

Animals↗

p53 overexpression in the multistep process of esophageal carcinogenesis.

The timing of p53 mutation in the multistep process of esophageal carcinogenesis is still under debate. We tested p53 expression in 16 samples of low-grade and 29 samples of high-grade esophageal dysplasia (ED) coexisting with esophageal squamous cancer (ESC) in 31 patients who underwent total esophagectomy. In normal mucosa, a positive immunoreaction was detected in 10 of 31 cases, always restricted to the lower half of the epithelial thickness. We detected p53-positive nuclei in 11 of 16, 23 of 29, and 23 of 31 samples of low-grade ED, high-grade ED, and ESC, respectively. Cases exhibiting positive staining in dysplastic samples also demonstrated positive immunoreaction in the carcinomatous tissue. Immunoreactivity in cancer cells was never found in the absence of positive dysplastic nuclei. A significantly higher score of immunoreactive nuclei was detected in high-grade versus low-grade and in low-grade compared with normal mucosa. These data suggest that p53 mutation may represent an early event in esophageal oncogenesis.

Adult↗

Determinants for the development of chronic atrophic gastritis and intestinal metaplasia in the stomach.

This study aimed to identify the factors associated with the development of gastric precancerous changes, in a prospective series of patients undergoing endoscopy. Risk factors and associated mucosal changes were evaluated in 134 endoscoped patients affected by chronic non-atrophic (n = 76) or atrophic gastritis (CAG) (n = 32), with or without intestinal metaplasia (IM), or lacking any major histological changes (n = 26). The following variables were taken into account: age, alcohol consumption, smoking habit, vitamin C intake (using a questionnaire), gastric juice vitamin C (HPLC on gastric juice samples obtained at endoscopy), H. pylori infection, gastric mucosa malondialdehyde (MDA; a measure of free radical production) and extent of CAG in gastric biopsies (only for IM). Tissue MDA levels were significantly higher, and vitamin C levels significantly lower in CAG and IM patients (P = 0.01). Multiple regression analysis showed significant correlations for: CAG vs age (P < 0.02), MDA (< 0.02) and gastric vitamin C (< 0.05); IM vs age (P < 0.0005), CAG (< 0.0005) and MDA (< 0.001). Using stepwise discrimination analysis, the independent variables included in the model were: for CAG, age (P < 0.003), MDA (< 0.05), gastric juice vitamin C (< 0.05); for IM, CAG (P < 0.0005), age (< 0.001), MDA (< 0.03) and vitamin C intake (< 0.05). H. pylori was not included. The major determinants for CAG and IM were age, free radical production (as measured by MDA), vitamin C (for CAG) and vitamin C intake and CAG (for IM).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

HLA-DR expression in liver in human HIV infection.

In order to clarify the relationship between HLA-DR antigen expression in the liver and human HIV infection, we studied 66 non-autopsy specimens obtained from HIV-infected patients with and without AIDS; 33 biopsies from HIV-negative patients were considered for control purposes. All biopsies were immunohistochemically tested with monoclonal antibody TAL-1B5, reacting with the alpha-chain subunit of HLA-DR in the avidin biotin complex method. We found occasional and usually weak HLA-DR expression on hepatocytes and biliocytes. By comparison with controls, HIV-positive cases showed a significantly higher percentage of HLA-DR immunoreactivity on hepatic artery and portal vein endothelial cells. No relationship was demonstrated between HLA-DR endothelial expression and the CD4+ lymphocyte count or grade of portal/periportal inflammation in the liver. The biological significance of HLA-DR expression on intrahepatic vascular endothelial cells in HIV infection still remains to be clarified.

Adult↗