Presence of Chlamydia pneumoniae in abdominal aortic aneurysms is not associated with increased activity of matrix metalloproteinases.

OBJECTIVE: to test the hypothesis that the presence of Chlamydia pneumoniae (C. pneumoniae) in the wall of abdominal aortic aneurysms (AAA) is associated with increased activity of matrix metalloproteinase (MMP)-2 and/or MMP-9. DESIGN: case-control study. MATERIAL AND METHODS: in a series of 40 patients with AAA > or =5cm in maximal cross-sectional diameter, C. pneumoniae-DNA was identified in the aneurysm wall by nested PCR in 14 (35%) patients. Another 14 C. pneumoniae-DNA-negative AAA patients from the same series, matched for gender and aneurysm diameter, were used as controls. In each group there were 7 asymptomatic (aAAA) and 7 ruptured (rAAA) aneurysms. MMP-2 and -9 activity was estimated in AAA wall biopsies by gelatin zymography. RESULTS: patients with a C. pneumoniae-DNA-positive aneurysm wall specimen showed an over-all lower activity of MMP-2 and MMP-9 (pro- and active enzyme) compared to the C. pneumoniae-DNA negative patients. However, there were no statistically significant differences in MMP activity between the two groups of patients with aAAA. Among patients with rAAA both pro-MMP-9 (p=0,026) and active-MMP-9 (p=0.007) were significantly lower in C. pneumoniae-DNA-positive patients compared to C. pneumoniae-DNA-negative patients, whereas there were no significant differences in pro-MMP-2 or active-MMP-2. CONCLUSION: this preliminary study does not support the hypothesis that the presence of C. pneumoniae in the AAA wall is associated with increased activity of MMP-2 and MMP-9.

In vitro degradation of aortic elastin by Chlamydia pneumoniae.

OBJECTIVES: to investigate whether Chlamydia pneumoniae (C. pneumoniae) may increase elastin degradation in the aortic wall. MATERIALS AND METHODS: eighteen full thickness aortic wall samples from non-aneurysmal infrarenal abdominal aortas were collected from autopsies. Two adjacent and equally large pieces were cut out of each aortic sample. From each sample, one piece was incubated in a HEp-2 cell culture infected with C. pneumoniae and the other piece was incubated in an uninfected HEp-2 cell culture. The incubation time was one week at 35 degrees C. The concentration of elastin-derived peptides (EDP) (ng/ml) in the medium of each cell culture was measured in duplicate. For each paired sample, delta-EDP (EDP in HEp-2 cell culture infected with C. pneumoniae- EDP in uninfected HEp-2 cell culture) was calculated. RESULT: there was a significantly increased degradation of aortic elastin, estimated by EDP concentrations in cell culture conditioned medium, when aortic wall samples were incubated in C. pneumoniae cultures compared with uninfected cultures (p=0.025, Wilcoxon signed ranks test). CONCLUSION: these results indicate that there is a relationship between the presence of C. pneumoniae and increased elastin degradation in the aortic wall in vitro. This suggests C. pneumoniae in the aortic wall directly or indirectly leads to the degradation of aortic elastin.

A preliminary evaluation of a new enzyme immunoassay to detect Chlamydia pneumoniae-specific antibodies.

New enzyme immunoassays (EIAs) for determination of specific IgG, IgA, and IgM antibody titers to Chlamydia pneumoniae were evaluated independently in three research laboratories. Specificity of the EIAs was enhanced by removing LPS from the chlamydial antigen. The performance of these EIAs was evaluated in comparison with the microimmunofluorescence (MIF) test using specimens from: (i) a group of adult patients with community-acquired pneumonia (CAP) previously diagnosed as having an acute chlamydial infection by the complement fixation test or the whole inclusion fluorescence test; (ii) from a group of adult patients with acute respiratory tract infections; and (iii) from a group of young children consecutively presenting with acute respiratory tract infections. The MIF test and the EIAs detected acute infections in paired serum specimens from 12 of 14 patients from the first group. Eleven of these 12 patients were positive in both tests. The MIF test detected seven acute infections in single convalescence serum specimens from eight patients. Two of these were also positive in the EIAs. Paired serum specimens from the second group of adult patients (n=12) were collected during an epidemic of C. pneumoniae. The EIAs detected six acute infections. The MIF test detected two additional patients with acute infections. From the group of young children (n=30), the EIAs detected two patients with acute infections. Our conclusion from this preliminary evaluation is that these EIAs could be useful for laboratory diagnosis of acute C. pneumoniae infections. Comprehensive prospective studies should provide suitable data to calculate the sensitivity, specificity, and predictive values.

Small-bowel obstruction secondary to malignant disease: an 11-year audit.

OBJECTIVE: To determine the efficacy and long-term prognosis for operative versus nonoperative treatment of small-bowel obstruction (SBO) secondary to malignant disease. DESIGN: A chart review. SETTING: A university-affiliated teaching hospital. PATIENTS: The medical records of all patients with malignant disease as the established etiology of their obstruction who presented to the Sir Mortimer B. Davis-Jewish General Hospital, Montreal, between 1986 and 1996 were reviewed. There were 32 patients accounting for 74 admissions. INTERVENTIONS: Selective nonoperative management and exploratory laparotomy, immediate or delayed. MAIN OUTCOME MEASURES: The value of nonoperative management and need for operation. RESULTS: Colorectal and ovarian neoplasms were the principal primary malignant diseases that led to SBO. The median time between diagnosis of the malignant disease and SBO was 1.1 years. At their initial presentation, 80% of patients were treated by operation, but 47% of these patients had an initial trial of nonoperative treatment. Reobstruction occurred in 57% of patients who were operated on compared with 72% of patients who were not. The median time to reobstruction was 17 months for patients who underwent operation compared with 2.5 months for patients who did not. Also, 71% of patients were alive and symptom free 30 days after discharge from operative treatment compared with 52% after nonoperative treatment. Postoperative morbidity was 67%. Mortality was 13%, and 94% of patients eventually died from complications of their primary disease. CONCLUSIONS: SBO secondary to malignant disease usually indicates a grim prognosis. Operative treatment has better outcome than nonoperative management in terms of symptom free interval and reobstruction rates. However, it is marked by high postoperative morbidity. We recommend that, after short trial of nasogastric decompression, patients with obstruction secondary to malignant disease be operated on if clinical factors indicate they they will survive the operation.

Etiology of small bowel obstruction.

BACKGROUND: Small bowel obstruction (SBO) is a major cause of morbidity and financial expenditure in hospitals around the world. The leading cause of SBO in the western world has become adhesions. The goal of this study was to determine the causes of SBO. METHODS: The medical records of all patients admitted to one hospital between 1986 and 1996 with the diagnosis of SBO were reviewed retrospectively. This included 552 patients accounting for 1,001 admissions. RESULTS: The etiology of SBO was adhesions (74%), Crohn's disease (7%), neoplasia (5%), hernia (2%), radiation (1%), and miscellaneous (11%). Patients with Crohn's disease were younger than patients with other etiologies. Surprisingly, recurrence rates were similar for patients treated operatively as for those treated nonoperatively with the exception in the hernia group where higher recurrence rates were noted for patients initially treated in a nonoperative manner. CONCLUSION: The most common cause of SBO is adhesions followed by Crohn's disease and neoplasia.

Comparison of five serologic tests for diagnosis of acute infections by Chlamydia pneumoniae.

Serology is often used to diagnose acute infections by Chlamydia pneumoniae. In this study paired sera from patients with acute respiratory tract infection during an epidemic of C. pneumoniae infections were examined by five different antibody tests. These tests were the complement fixation (CF) test, the microimmunofluorescence (MIF) test, a recombinant enzyme immunoassay (rEIA) (Medac) based on a recombinant lipopolysaccharide of chlamydia and measuring antibodies to a common chlamydial antigen, and two tests that utilize preparations of C. pneumoniae organisms, the SeroCp-EIA (Savyon) (with preserved lipopolysaccharide) and the LOY-EIA (Labsystems) (without this antigen). Both of the last two tests should measure specific antibodies to C. pneumoniae, although cross-reacting antibodies may also be detected by the SeroCp-EIA. Acute infection of C. pneumoniae was serologically confirmed in 44% of the cases by at least two different tests. Using an expanded "gold standard," i.e., the presence of significant reactions in at least two tests, the sensitivity of the CF test was 69%, that of the MIF test was 88%, that of the rEIA was 89%, that of the LOY-EIA was 96%, and that of the SeroCp-EIA was 92%. Specificity was high for all methods, but adjustments of diagnostic criteria were made to several of the tests. The basis for these adjustments and supportive data are presented. Infections of C. pneumoniae were detected in patients from 8 to 83 years of age. Two peaks in the incidence of such infections were observed: one among young teenagers and a second in adults 30 to 45 years of age, corresponding to parents of young teen-agers. The tests were equally sensitive in different age groups. Reinfections seemed to be rare.

Natural history of patients with adhesive small bowel obstruction.

BACKGROUND: Small bowel obstruction (SBO) is a major cause of morbidity and financial expenditure. The goals of this study were to determine factors predisposing to adhesive SBO, to note the long-term prognosis and recurrence rates for operative and non-operative treatment, to elicit the complication rate of operations and to highlight factors predictive of recurrence. METHODS: The medical records of all patients admitted to one hospital between 1986 and 1996 with the diagnosis of SBO were reviewed retrospectively. This included 410 patients accounting for 675 admissions. RESULTS: The frequency of previous operation by procedure type was colorectal surgery (24 per cent), followed by gynaecological surgery (22 per cent), herniorrhaphy (15 per cent) and appendicectomy (14 per cent). A history of colorectal surgery (odds 2.7) and vertical incisions (odds 2.5) tended to predispose to multiple matted adhesions rather than an obstructive band. At initial admission 36 per cent of patients were treated by means of operation. As the number of admissions increased, the recurrence rate increased while the time interval between admissions decreased. Patients with an adhesive band had a 25 per cent readmission rate, compared with a 49 per cent rate for patients with matted adhesions (P<0.004). At the initial admission 36 per cent of patients were treated surgically. Patients treated without operation had a 34 per cent readmission rate, compared with 32 per cent for those treated surgically (P not significant), a shorter time to readmission (median 0.7 versus 2.0 years; P<0.05), no difference in reoperation rate (14 versus 11 per cent; P not significant) and fewer inpatient days over all admissions (4 versus 12 days; P<0.0001). CONCLUSION: The likelihood of reobstruction increases and the time to reobstruction decreases with increasing number of previous episodes of obstruction. Patients with matted adhesions have a greater recurrence rate than those with band adhesions. Non-operative treatment for adhesions in stable patients results in a shorter hospital stay and similar recurrence and reoperation rates, but a reduced interval to reobstruction when compared with operative treatment.

Touchdown enzyme time release-PCR for detection and identification of Chlamydia trachomatis, C. pneumoniae, and C. psittaci using the 16S and 16S-23S spacer rRNA genes.

Three touchdown enzyme time release (TETR)-PCR assays were used to amplify different DNA sequences in the variable regions of the 16S and 16S-23S spacer rRNA genes specific for Chlamydia trachomatis, Chlamydia pneumoniae, and Chlamydia psittaci as improved tests for sensitive diagnosis and rapid species differentiation. The TETR-PCR protocol used 60 cycles of amplification, which provided improved analytical sensitivity (0.004 to 0.063 inclusion-forming unit of Chlamydia species per PCR). The sensitivity of TETR-PCR with primer set CTR 70-CTR 71 was 96.7%, and the specificity was 99.6%, compared to those of the AMPLICOR PCR for the detection of C. trachomatis in vaginal swab samples. TETR-PCR for C. pneumoniae with primer set CPN 90-CPN 91 was 90% sensitive and 93.3% specific compared with a nested PCR with primer set CP1/2-CPC/D for clinical respiratory samples. TETR-PCR for C. psittaci with primer set CPS 100-CPS 101 showed substantial agreement with cell culturing (kappa, 0.78) for animal tissue samples. Primer sets were then combined into a single multiplex TETR-PCR test. The respective 315-, 195-, and 111-bp DNA target products were precisely amplified when DNA from each of the respective Chlamydia species or combinations of them was used. Multiplex chlamydia TETR-PCR correctly identified one strain of each of the 15 serovars of C. trachomatis, 22 isolates of C. pneumoniae, and 20 isolates of C. psittaci. The primer sets were specific for each species. No target products were amplified when DNA from C. pecorum or a variety of other microorganisms was tested for specificity. TETR-PCR with primers selected for specific sequences in the 16S and 16S-23S spacer rRNA genes is a valuable test that could be used either with individual primers or in a multiplex assay for the identification and differentiation of Chlamydia species from culture isolates or for the detection of chlamydiae in clinical samples.

Chlamydia pneumoniae DNA detection in peripheral blood mononuclear cells is predictive of vascular infection.

Abdominal aortic aneurysm tissue and peripheral blood mononuclear cells (PBMC) of 41 consecutive subjects undergoing abdominal aortic aneurysm surgery were analyzed by polymerase chain reaction (PCR) for the presence of Chlamydia pneumoniae, Mycoplasma pneumoniae, and Helicobacter pylori DNA. Twenty patients (49%) were positive for C. pneumoniae DNA-16 (39%) in both PBMC and aneurysm tissue, 3 (7.3%) in PBMC only, and 1 (2.4%) in the artery specimen only. Previous exposure to C. pneumoniae was confirmed in 19 (95%) of the 20 PCR positive subjects by C. pneumoniae-specific serology, using the microimmunofluorescence test. None was positive for H. pylori or M. pneumoniae DNA, either in the PBMC or in the artery specimens. In conclusion, carriage of C. pneumoniae DNA is common both in PBMC and in abdominal aortic tissue from patients undergoing abdominal aneurysm surgery. Blood PCR may be a useful tool for identifying subjects carrying C. pneumoniae in the vascular wall.

Chlamydia pneumoniae in human abdominal aortic aneurysms.

OBJECTIVES: To investigate the presence of Chlamydia pneumoniae DNA in the wall of infrarenal abdominal aortic aneurysms, and in the wall of non-aneurysmal infrarenal abdominal aortas. DESIGN: Case-control study. MATERIALS AND METHODS: The study group consisted of 40 patients operated transperitoneally for an infrarenal abdominal aortic aneurysm (IAAA) (eight females, 32 males; mean age 69 years, median age 68 years). Specimens from the aneurysm wall were taken peroperatively under sterile conditions. The control group consisted of 40 deceased persons without aortic aneurysms (14 females, 26 males; mean age 71 years, median age 70 years). Specimens from the non-aneurysmal infrarenal aortas (NIAA) were collected within 48 h after death. The specimens from both groups were frozen at -70 degrees C immediately after collection. A nested polymerase chain reaction (PCR) method, using two sets of primers designed to detect a fragment of the major outer membrane protein gene of C. pneumoniae, was used. RESULTS: The detection of C. pneumoniae-specific DNA was significantly higher in the study group (14/40 = 35%) than in the control group (2/40 = 5%); (p = 0.001). No clinical factor predicting the presence of C. pneumoniae in the aneurysm wall, could be found. CONCLUSION: Chlamydia pneumoniae was detected at a significantly higher frequency in the wall of IAAAs than in the wall of NIAAs. Although this finding does not prove that C. pneumoniae causes IAAAs, further studies on the possible role of C. pneumoniae in the pathogenesis of aneurysms should be performed.

Sero-epidemiological association between human-papillomavirus infection and risk of prostate cancer.

Some epidemiological studies of prostate cancer have suggested the existence of a sexually transmitted risk factor, and some studies have reported the presence of human papillomavirus (HPV) DNA in prostate-cancer tissue. To perform a sero-epidemiological evaluation of whether HPV infection is associated with increased risk for prostate cancer, we performed a nested case-control study within a serum bank containing samples from 20,243 healthy Finnish men. We identified 165 cases of prostate cancer that were diagnosed up to 24 years after donation of the serum sample. Two control subjects per case were selected, matched for gender, age and municipality of residence. Serum samples were analyzed for the presence of IgG antibodies against 4 HPV types and against Chlamydia. The presence of antibodies against HPV type 18 was associated with a 2.6-fold increased risk of developing prostate cancer during follow-up (p < 0.005). HPV type 16 tended to be associated with subsequent prostate-cancer occurrence (relative risk: 2.4, p = 0.06), whereas seropositivity for HPV type 11 or type 33 or for Chlamydia was not associated with risk. The results suggest that infection with oncogenic HPV might be involved in the etiology of a minority of prostate cancers.

Rapid diagnosis of respiratory Chlamydia pneumoniae infection by nested touchdown polymerase chain reaction compared with culture and antigen detection by EIA.

Chlamydia pneumoniae is a common cause of respiratory tract infection and community-acquired pneumonia. During an extensive outbreak of C. pneumoniae in northern Sweden, 319 respiratory samples from 129 persons were collected. Sputum, throat, and nasopharyngeal samples were obtained and analyzed by nested touchdown polymerase chain reaction (PCR), EIA, and culture in Hep-2 and McCoy cells. Serology was performed by complement fixation and microimmunofluorescence tests. By PCR, 30 patients were diagnosed with C. pneumoniae compared with 26 positive by EIA and 23 by culture. The finding of C. pneumoniae in the respiratory samples was accompanied by serology indicating acute infection in 26 (96%) of 27 patients for whom adequate sera were available. Nested PCR was sensitive and reliable for diagnosing acute respiratory C. pneumoniae infection. Sputum samples had the highest diagnostic efficacy, and the nested type of PCR was superior to one-step PCR. EIA and culture were less sensitive than nested PCR.

Intravesical therapy with adriamycin in patients with superficial bladder tumors.

An 80-mg dose of adriamycin was given intravesically once a month to 58 patients with superficial transitional cell tumor of the bladder. Complete cytological remission was achieved in all 11 patients with previously untreated, flat carcinoma in situ lesions. In 19 carcinoma in situ patients, who were previously treated by other means, cytological remission occurred in 74%. In T1 and T2 papillary tumor remission occurred in 65 and 20%, respectively. Minor local side effects were observed.

Estramustine versus conventional estrogenic hormones in the initial treatment of highly or moderately differentiated prostatic carcinoma. A randomized study.

In a prospective randomized multicenter trial patients with highly or moderately differentiated prostatic carcinoma, previously untreated, were allotted either to oral Estramustine phosphate or to intramuscular polyestradiol phosphate plus oral ethinyl estradiol. As regards frequency and duration of tumour remission there was no statistical difference between the two groups. Nor did they differ significantly with respect to adverse reactions. This is an interim report and will be followed later on by a final evaluation.