Programmes to reduce pelvic inflammatory disease--the Swedish experience.
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Biomedical subjects
Publications and source records attributed to D Danielsson.
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OBJECTIVES: To study organ involvement, anti-neutrophil cytoplasmic antibodies (ANCA) patterns, trends in yearly frequencies and seasonal variations of symptom onset in patients hospitalized because of small vessel vasculitides during a 21 year period (1975-95). DESIGN: A retrospective investigation was conducted of 138 patients hospitalized with a diagnosis of small vessel vasculitides, as defined by the Chapel Hill Consensus Conference, within the County of Orebro, a mixed urban and rural area of central Sweden. SETTING: Orebro Medical Center Hospital, Orebro, Sweden and two district hospitals within the County of Orebro, Sweden. RESULTS: During the studied period there were 19 patients with a diagnosis of Wegener's granulomatosis (WG), 70 patients with microscopic polyangiitis (MPA), 36 patients with renal limited vasculitis (RLV), two with Churge-Strauss vasculitis (C-S), seven with Henoch-Schönleins purpura (HSP) and four with essential cryoglobulinemic vasculitis (ECV). Renal involvement was present in 123 patients (89.1%). A positive c- and/or pANCA was found in nearly 90% of the 111 patients where sera were available. Calculations of frequency data, restricted to the primary catchment area for patients with ANCA associated vasculitis and renal involvement (WG, MPA, RLV) during a 21-year period (1975-95) gave a mean annual frequency of 1.6 per 100,000 adults (95% CI: 1.2-3.1); for this group of patients with the inclusion of those with C-S, HSP and ECV during the last 10 year period (1986-95) gave a mean annual frequency of 2.5 per 100,000 adults (95% CI: 1.7-3.4), for male adults 3 per 100,000 (95% CI: 1.6-4.4), and female adults 1.9 (95% CI: 0.9-2.8). A frequency peak of 6.3 per 100,000 was seen for men aged 55-64. A periodic fluctuation of the frequencies with peaks every 3-4 years was noted for patients with ANCA related vasculitis (WG, MPA, RLV) during the 21-year period 1975-95. Onset of symptoms was predominantly noticed during the winter months (December-February) for patients with a positive cANCA. CONCLUSION: The observed frequencies in our study of patients with small vessel vasculitides were higher than those previously documented. We also showed a periodic fluctuation of the annual frequencies and a seasonal variation of symptom onset.
BACKGROUND: Perinuclear antineutrophil cytoplasmic antibodies (P-ANCA) are found in 48%-83% of serum samples from patients with ulcerative colitis (UC). Their pathogenic role and initiating stimuli are unknown. In contrast to patients with vasculitides and ANCA reactivities, the antibodies in UC patients do not react with myeloperoxidase (MPO) or proteinase 3 (PR3). The aim of the present study was to investigate whether bacterial species of the intestinal tract and other sources could interfere with P-ANCA in sera from patients with UC. METHODS: Seventeen P-ANCA-positive and anti-MPO-negative serum samples from patients with UC were tested with Escherichia coli 014 and Staphylococcus aureus Wood 46. Six of these serum samples with different P-ANCA titres were selected to test further the influence of 15 different gram-negative or gram-positive bacterial strains. Six anti-MPO positive P-ANCA, 5 anti-PR3 positive C-ANCA, and 10 antinuclear antibody (ANA)-positive serum samples were used as controls. The antineutrophil cytoplasmic antibodies (ANCAs) were analysed by an indirect immunofluorescence method (IIF) on ethanol-fixed neutrophils, and the ANAs were tested by IIF on HEp-2 cells or rat liver tissues. The bacteria used in the experiments were either live or killed by formalin or glutaraldehyde fixation or heated at 80 degrees C for 30 min. The test was first performed as a bacterial absorption test with sedimented organisms and then at various temperatures with the supernatant from suspension of live bacteria. RESULTS: Both MPO-positive and MPO-negative P-ANCA reactivity was abolished by absorption of patient sera with live E. coli and Proteus mirabilis but not with bacteria representing members of 10 other species, suggesting that antibody reactivity was absorbed away. However, continued experiments indicated that the inhibition of P-ANCA was not due to classic antigen-antibody interactions but rather to decomposition of the antigenic substrate of the neutrophils by factors present in the supernatants of live E. coli and P. mirabilis. The activity of the supernatant was temperature-dependent, with strong activity at room temperature and 37 degrees C, no activity at 0 degrees C, and abolished by mild heat treatment (56 degrees or 60 degrees C). No activity was shown in the supernatants from bacteria treated with formaldehyde or glutaraldehyde. CONCLUSIONS: Soluble material from live E. coli and P. mirabilis has the capacity to decompose the antigenic substrate of neutrophils responsible for both MPO-positive and MPO-negative P-ANCA, most probably brought about through enzymatic activity. Anti PR3-positive C-ANCA were not affected, which suggests substrate specificity of the proposed enzymatic activity.
The possible role of glycosphingolipids as adhesion receptors for the human gastric pathogen Helicobacter pylori was examined by use of radiolabeled bacteria, or protein extracts from the bacterial cell surface, in the thin-layer chromatogram binding assay. Of several binding specificities found, the binding to lactosylceramide is described in detail here, the others being reported elsewhere. By autoradiography a preferential binding to lactosylceramide having sphingosine/phytosphingosine and 2-D hydroxy fatty acids was detected, whereas lactosylceramide having sphingosine and nonhydroxy fatty acids was consistently nonbinding. A selective binding of H. pylori to lactosylceramide with phytosphingosine and 2-D hydroxy fatty acid was obtained when the different lactosylceramide species were incorporated into liposomes, but only in the presence of cholesterol, suggesting that this selectivity may be present also in vivo . Importantly, lactosylceramide with sphingosine and hydroxy fatty acids does not bind in this assay. Furthermore, a lactosylceramide-based binding pattern obtained for different trisaccharide glycosphingolipids is consistent with the assumption that this selectivity is due to binding of a conformation of lactosylceramide in which the oxygen of the 2-D fatty acid hydroxyl group forms a hydrogen bond with the Glc hydroxy methyl group, yielding an epitope presentation different from other possible conformers. An alternative conformation that may come into consideration corresponds to the crystal structure found for cerebroside, in which the fatty acid hydroxyl group is free to interact directly with the adhesin. By isolating glycosphingolipids from epithelial cells of human stomach from seven individuals, a binding of H.pylori to the diglycosylceramide region of the non-acid fraction could be demonstrated in one of these cases. Mass spectrometry showed that the binding-active sample contained diglycosylceramides with phytosphingosine and 2-D hydroxy fatty acids with 16-24 carbon atoms in agreement with the results related above.
Some clinical isolates of nonopsonized H. pylori have the ability to activate neutrophils to an oxidative burst (neutrophil activating capacity, NAC), and such strains were significantly more often isolated from patients with peptic ulcer disease and active chronic gastritis. The purpose of the present work was to investigate the effect of rebamipide (Mucosta) on the release of reactive oxygen metabolites from neutrophils activated by various strains of H. pylori with or without NAC, nonopsonized or opsonized, using as controls fMLP and PMA, known activators of neutrophils, and to study the kinetics of these events by luminol-enhanced chemiluminescence and by flow cytometry. The results showed that the oxidative burst induced in neutrophils by fMLP and by nonopsonized or opsonized H. pylori with NAC was inhibited by rebamipide in a dose-dependent manner both in the early and late phases of activation. In contrast, the oxidative burst induced by opsonized H. pylori without NAC was not inhibited by rebamipide, which might indicate that it does not have the ability to block CR1 or CR3 receptors involved in opsonic phagocytosis but still has the ability to block the receptor(s) for NAC. The oxidative burst induced by PMA, which primarily activates protein kinase C, was not inhibited in the early phase but diminished 40-45% in the late phase with the 2 mM concentration of rebamipide, probably due to scavenging of reactive oxygen species. In conclusion, rebamipide has the ability to diminish the oxidative burst of neutrophils activated by nonopsonized or opsonized H. pylori organisms with neutrophil activating capacity, most likely through the blocking of fMLP-related receptors, inhibition of the production of reactive oxygen species, and the scavenging of such metabolites. Rebamipide may therefore be useful to prevent gastroduodenal lesions associated with gastric mucosal inflammation in H. pylori infection.
The possible interaction of the neutrophil-activating protein of Helicobacter pylori with target cell glycoconjugates was investigated by the binding of 125I-labeled recombinant protein to glycosphingolipids from human neutrophils in solid phase assays. Thereby, a distinct binding of the neutrophil-activating protein to four bands in the acid glycosphingolipid fraction from human neutrophils was detected, whereas no binding to the non-acid glycosphingolipids or polyglycosyl ceramides from these cells was obtained. When using glycosphingolipids not present in the cell membrane of human neutrophils, it was found that the neutrophil-activating protein also bound to sulfated glycosphingolipids as sulfatide and sulfated gangliotetraosyl ceramide. Comparison of the binding preferences of the protein to reference glycosphingolipids from other sources suggested that in human granulocytes, the neutrophil-activating protein of H. pylori preferentially recognizes glycoconjugates with a terminally unsubstituted NeuAcalpha3Galbeta4GlcNAcbeta3Galbeta4GlcNAcbeta sequence.
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BACKGROUND/AIMS: Mucosal adherent bacterial flora in chronic alcoholics was studied and compared to a control group referred for upper endoscopy, mainly for dyspepsia. METHODS: 22 alcoholics, admitted to hospital for detoxification, were examined using upper gastrointestinal endoscopy. Gastric and duodenal biopsies were taken for tissue pathology, quantitative and qualitative anaerobic and aerobic bacteriological culture and for culture of Helicobacter pylori (antral biopsies). 12 nonalcoholics, admitted for upper endoscopy mainly for dyspepsia, were chosen as a control group. Seven of these had used gastric acid inhibitors. RESULTS: Gastrointestinal symptoms were common among alcoholics: 20/22 (90%) had diarrhea, nausea and/or abdominal pain. There were signs of gastritis by endoscopy in 64% of the alcoholics and in 58% of the controls. Tissue pathology, however, showed active chronic antral gastritis in 27% of the alcoholics and in 42% of the controls. H. pylori were isolated in 7/22 of the alcoholics and in 4/12 of the controls, which corresponds to the mean prevalence for these age groups in Sweden. Significantly more bacteria, dominated by gram-positive aerobic cocci, were present in the gastric biopsies of alcoholics than in those of controls (mean of 2.9 x 10(6)/g material versus 4.4 x 10(5), p < 0.05). There were 2.6 times more bacteria in the duodenal biopsies of alcoholics than in those of the controls (p > 0.05, NS). Bacterial overgrowth (defined as >2 x 10(3) organisms/g material) was found in the stomach in 20/22 (90%) alcoholics and in 6/12 (50%) controls (p < 0.01). CONCLUSION: Alcoholics have an increased frequency of bacterial overgrowth in the upper gastrointestinal tract. This may contribute to the common gastrointestinal symptoms.
OBJECTIVE: To study some mechanisms involved in Helicobacter pylori (H. pylori)-induced gastric carcinogenesis. DESIGN: In vitro study. SETTING: Medical centre hospital, Sweden. INTERVENTIONS: Mutagenicity in Ames' test of neutrophils challenged for 2 hours or more by two different strains of H. pylori. One strain designated NCTC 11637 by the National College of Type Cultures activated neutrophils to an oxidative burst and producing vacuolating cytotoxin, the other strain C-7050 lacked these abilities. Mutagenicity was also studied with sterile human gall bladder bile alone added to neutrophils or in combination with both neutrophils and H. pylori. RESULTS: There was no increase in the number of revertants with the crude suspension or the supernatant of neutrophils challenged for 1 hour or less with H. pylori, bile, or the combination of both. However, in 5 out of 19 experiments there was significant mutagenicity after challenge of neutrophils for 2 hours or more with either strain of H. pylori, bile, or the combination of the two. The strongest mutagenicity was obtained after challenge over night (18 hours) with the combination of H. pylori and bile. CONCLUSION: Mutagenicity occurs when neutrophils are challenged with H. pylori and bile. Factors other than reactive oxygen metabolites seem to be responsible.
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BACKGROUND: The aetiology and pathogenesis of collagenous colitis are unknown. Autoimmunity has been suggested, but no serological findings have supported such a theory. AIMS AND METHODS: Serum from 38 collagenous colitis patients and 38 matched healthy controls was analysed for autoantibodies--that is, antinuclear antibodies, antineutrophil cytoplasmic antibodies, smooth muscle and mitochondrial antibodies, rheumatoid factor and antibodies to thyroglobulin and microsomal antigen, together with antibodies to endomysium, gliadin, and cardiolipin. The serum values of IgA, IgG, IgM, and IgG-subclasses, and complement factors C3 and C4 were also determined. RESULTS: In patients with collagenous colitis the mean value of IgM was significantly increased 2.5 g/l (95% CI; 1.9, 3.2) compared with 1.4 g/l (95% CI; 1.2, 1.7) in controls (p = 0.002). Antinuclear antibodies occurred in nine of 38 patients compared with three of 38 controls, this difference was not statistically significant (p = 0.11). The results of all other immunoglobulins, complement factors, and specific antibodies showed no statistical difference between patients and controls. CONCLUSIONS: No firm evidence for an autoimmune genesis in collagenous colitis is found in this study, although the findings of a positive ANA-titre in some patients and an increased IgM level might give some support for this hypothesis.
BACKGROUND: It has been shown that perinuclear antineutrophil cytoplasmic antibody (P-ANCA) may be associated with pouchitis after proctocolectomy with ileal pouch-anal anastomosis (IPAA) for ulcerative colitis (UC). METHODS: P-ANCA was studied with the indirect immunofluorescence technique in 76 UC patients after IPAA. Twenty-eight patients had had pouchitis, whereas 48 patients did not. RESULTS: P-ANCA was found in 49 of 76 (64.5%) UC patients after IPAA. In patients who had had pouchitis attacks within 1 year of serum sampling (group 1) 12 of 12 (100%) patients had positive P-ANCA. In patients who had had pouchitis attacks 1 or several years before the serum sampling (group 2) 9 of 16 (56.3%) patients had positive P-ANCA. In patients who had not yet had a pouchitis attack (group 3) 28 of 48 (58.3%) were positive. The occurrence of P-ANCA in group 1 was significantly higher than in group 2 (p = 0.01) or group 3 (p = 0.005). No statistically significant difference was found between the occurrence of P-ANCA in group 2 and group 3. Furthermore, we found that the titres of P-ANCA in the pouchitis patients were associated with the observation time since the first pouchitis attack to the time of serum sampling (r = -0.43, p = 0.02) and a pouchitis relapse index (average pouchitis attacks per year, r = 0.47, p = 0.03). CONCLUSIONS: P-ANCA was found in UC patients after proctocolectomy with IPAA. Patients with recent (< or = 12 months) or ongoing pouchitis are all P-ANCA-positive. Pouchitis patients with higher P-ANCA titres are more prone to have frequent relapses.
BACKGROUND: Some Helicobacter pylori strains activate human neutrophils without opsonins and/or produce vacuolating cytotoxin. METHODS: Human gastric isolates of H. pylori were studied for their ability to nonopsonized induce an oxidative burst in human neutrophils as measured by chemiluminescence and for the production of vacuolating cytotoxin. In all, 80 strains were examined, and the type and grade of inflammation in the gastric biopsy specimens from the antrum and corpus of these patients were assessed in accordance with the Sydney system. RESULTS: CL+ (rapid, strong response in chemiluminescence) strains (p < 0.0001) and Tox+ (cytotoxin-producing) strains (p < 0.0001) were associated with higher acute inflammation scores in gastric ulcer patients. CL+ (p = 0.0002) and Tox+ (p < 0.0001) strains were also associated with higher chronic inflammation scores in gastric ulcer patients. CONCLUSIONS: CL+ and Tox+ strains seem to cause more severe inflammation in the gastric mucosa during H. pylori infection.
Eight clinical isolates and two reference strains of Helicobacter pylori were studied with regard to their interactions with HeLa cells. All the isolates adhered poorly to HeLa cells and the number of invasive bacteria was very low. No correlation was found between the adherence and invasiveness of the isolates on one hand, and the corresponding patients having ulcer or non-ulcer disease, or the ability of the strains to produce cytotoxin and to induce an oxidative burst of human polymorphonuclear leukocytes without opsonins, on the other. These results indicate that invasion of epithelial cells would play no important role in the pathogenesis of infections caused by H. pylori.
Perinuclear antineutrophil cytoplasmic antibodies (P-ANCA) have been demonstrated in patients with ulcerative colitis and in a higher frequency than expected in their first degree relatives. A hypothesis was proposed that P-ANCA is genetically determined and may represent a subclinical marker of genetic susceptibility to ulcerative colitis. This study analysed P-ANCA in monozygotic twins with inflammatory bowel disease to evaluate this hypothesis further. P-ANCA was analysed with indirect immunofluorescence technique in 12 monozygotic twin pairs with ulcerative colitis and 14 twin pairs with Crohn's disease. Furthermore, the study included 21 non-twin patients with ulcerative colitis, 18 non-twin patients with Crohn's disease, and 52 healthy controls matched for sex and age. In ulcerative colitis P-ANCA occurred in nine of 14 (64.3%) monozygotic twins and in 13 of 21 (61.9%) non-twin cases, which was significantly different compared with healthy controls who were positive in three of 52 (5.8%) cases (p < 0.0001). P-ANCA was found in two of 10 (20%) healthy twin siblings to twins with ulcerative colitis, which was not significantly different from healthy controls (p = 0.18). The results in Crohn's disease did not differ from healthy controls. Previous findings of P-ANCA occurring in ulcerative colitis but not in Crohn's disease are supported. This study does not support the hypothesis that P-ANCA is a subclinical marker of genetic susceptibility to ulcerative colitis.
As evidenced by electron microscopy, unopsonised bacteria of some Helicobacter pylori strains were readily taken into cytoplasmic vacuoles of human neutrophils; those of other strains were only rarely absorbed. The strains engulfed like this also induced strong oxidative burst reactions in neutrophils, as measured by chemiluminescence. This has been associated with peptic ulcer disease, in the past. The ultrastructural interaction patterns agreed with the reaction patterns shown by chemiluminescence.
Fifty-four clinical isolates of Helicobacter pylori from human gastric biopsy specimens were tested for cytotoxin production, as determined by intracellular vacuolization of HeLa cells, and for the induction of oxidative burst in human polymorphonuclear leukocytes (PMNLs), as measured by luminol-enhanced chemiluminescence (CL). Nonopsonized, 20 of the H. pylori strains induced a rapid and strong CL response, in contrast to 30 other strains, which showed only weak and slow responses. Another four strains gave inconclusive results. Cytotoxin production was demonstrated in 10 of the 20 strains with rapid responses but only in 3 of the 30 strains with slow and low responses (p = 0.0027, Fisher's exact test, two-tailed). Eleven of the 15 cytotoxin-producing strains (p = 0.0135) and 13 of the 20 strains with strong CL responses (p = 0.0209) were from 22 patients with peptic ulcer disease (PUD). The ability of some nonopsonized H. pylori to activate PMNLs showed co-variation with their ability to produce cytotoxin, but these two properties seem to be independent markers of PUD.
The diagnostic sensitivity and specificity of IgA endomysium antibodies (EmA) and IgA gliadin antibodies (AGA) for coeliac disease (CD) were studied in Swedish children. Indirect immunofluorescence was used for demonstration of EmA and the diffusion-in-gel enzyme-linked immunosorbent assay method for AGA. Serum samples were collected and analysed from 77 consecutive children undergoing small-intestinal biopsy on recognized clinical indications. The diagnostic sensitivity for CD was 98.1% for EmA and 86.5% for AGA. The specificity of the two tests was 92.7% for EmA and 92.7% for AGA in paediatric patients. In addition, 115 sera from control children showed 2.6% positive for EmA and 0.9% positive for AGA.