Search

Background: The proportion of Mycoplasma pneumoniae isolates carrying point mutations in the 23S region of the genome associated with macrolide resistance has increased. We evaluated the probability of developing M. pneumoniae macrolide resistance mutations during macrolide treatment. Methods: M. pneumoniae strains from initial and follow-up oropharyngeal samples from 38 Swedish patients were tested for 23S rRNA mutations by amplifying and pyrosequencing the gene target region. The duration of symptoms and of M. pneumoniae carriage were recorded in cases with and without the macrolide resistance mutation. Results: Macrolide resistance mutations were absent in all baseline strains. Twenty-two patients were prescribed macrolides; the longest M. pneumoniae carriage time in these patients was 7 months and the longest symptomatic period was 7.5 months. Macrolide resistance was detected after macrolide treatment in 1 patient (carriage time 3.5 months and symptomatic period 7.5 months). Conclusions: M. pneumoniae may develop macrolide resistance during macrolide treatment. A long duration of M. pneumoniae carriage and symptoms is common in patients regardless of the presence of resistance. (Less)

The synergism of infection with conventional cardiovascular risk factors in atherosclerosis is much debated. We hypothesized that coronary arterial injury correlates with infection recurrence and pathogen burden and is further aggravated by hypercholesterolemia. Forty-two Göttingen minipigs were assigned to repeated intratracheal inoculation of PBS, Chlamydia pneumoniae (Cpn), or both Cpn and influenza virus at 8, 11, and 14 wk of age. Animals were fed either standard or 2% cholesterol diet (chol-diet). At 19 wk of age coronary vasomotor responses to acetylcholine (ACh) and adenosine were assessed in vivo and blood and tissue samples were collected. Nonparametric tests were used to compare the groups. In cholesterol-fed animals, total cholesterol/HDL was significantly increased in infected animals compared with noninfected animals [3.13 (2.17–3.38) vs. 2.03 (1.53–2.41), respectively; P = 0.01]. C-reactive protein (CRP) rose in infected animals [10.60 (4.96–18.00) vs. 2.47 (1.44–3.01) μg/ml in noninfected; P < 0.01] without significant difference between the mono- and coinfected groups. Among coinfected animals, both CRP and haptoglobin were lower in those fed chol-diet than in those fed standard diet ( P < 0.05). The vasoconstricting response to ACh was most prominent in coinfected animals {769.3 (594–1,129) cm; P = 0.03 vs. noninfected [342 (309–455) cm] and P = 0.07 vs. monoinfected [415 (252.5–971.8) cm]}. Among monoinfected animals, similar to CRP, a trend for less vasoconstriction was observed in those fed chol-diet ( P = 0.08). Coinfection of piglets appears to be associated with more pronounced coronary muscarinic vasomotor dysfunction. In monoinfected animals, use of chol-diet seems to dampen both coronary dysfunction and systemic inflammation induced by infection.

Objectives: Endothelial cells of coronary arteries undergo apoptosis in patients with coronary artery disease. Carotid artery ultrasound studies show that after acute infections intima media thickness is increased. The possible effects of infections on endothelial cell apoptosis and the presence of apoptosis in early life remain unknown. Methods: Chlamydia pneumonia was inoculated three times at three weeks intervals to half of 32 piglets starting at the age of 8 weeks. Half of the piglets received cholesterol feeding. Morphological studies were done at the age of 19 weeks. Right coronary arteries were carefully excised and trimmed for excess tissue. The vessels were cut perpendicular to the long axis and sectioned at 5-μm thickness. TUNEL-assay was based on nick end labelling and staining of internucleosomal DNA fragments that are the biochemical hallmark of apoptosis. The percentage of apoptotic endothelial cells from intact endothe-lial cells was calculated using light microscopy. Results. Endothelial apoptosis appeared in 0.13% (SD 0.30%) of endothelial cells of non-infected piglets but in 0.75% (SD 0.62) of infected piglets (p

The Occurrence of plasma CRP during the febrile response after the first injection of penicillin was followed in 10 patients with early syphilis. An increase in CRP was noted after 12 hours, with a maximum after 24 hours. The appearance of this acute phase protein could not be correlated to cutaneous reaction, increased body temperature or leukocytosis, nor were baseline values of CRP correlated to clinical or serological activity of the infection. Patients with high levels of CRP prior to treatment suffered the most intense systemic reactions. No activation of the complement system during the Jarisch-Herxheimer reaction was detected from sequential determinations of C3, C4 and properdin.

Background. Mannose-binding lectin (MBL) is a multifunctional protein involved in innate immunity. We tested whether MBL and elevated viral and bacterial antibodies were risk factors for acute coronary events. Design. Controlled cohort study. Methods. A total of 354 patients with unstable angina pectoris (UA) or acute myocardial infarction (AMI) were compared with 334 paired controls. Results. Enterovirus titres were associated with increased risk of UA (odds ratio 10.04, P

Objectives: The role of persistent chlamydial infection of the Fallopian tubes in ectopic pregnancy is still unresolved. Therefore, we examined tissue of the Fallopian tubes from patients with ectopic pregnancy for the presence of Chlamydia trachomatis. In addition, other markers of C. trachomatis infection implicated in the pathogenesis of tubal damage were studied including antibodies to heat shock protein 60 of chlamydial and human origin. Study design: Fresh frozen tubal tissue from 55 patients with ectopic pregnancy in a hospital setting were examined for the presence of C. trachomatis DNA by polymerase chain reaction (PCR) and blood sample were analysed for antibodies to C. trachomatis including heat shock protein 60 (hsp60). Results: Chlamydial DNA was not detected in any of the 55 tubal specimens using a commercial test, Cobas Amplicor, Roche, and an in-house real time PCR able to detect a few copies of the organism. Logistic regression showed that chlamydial IgG antibodies were more common in a subgroup of patients with previous PID than in controls (OR = 7.84, CI 1.78-34.6). Specific antibodies to hsp60 of chlamydial (OR = 7.00, CI 1.50-32.6) but not of human origin (OR = 2.13, CI 0.14-31.6) were associated with ectopic pregnancy in this group. Conclusions: No evidence of persistent infection of C. trachomatis in the fallopian tubes at the time of ectopic pregnancy was found in this study. (C) 2006 Elsevier Ireland Ltd. All rights reserved. (Less)

A nested case–control study of invasive and in situ cervical cancer was performed within a community-based cohort of 13 595 Taiwanese women assembled in 1991, with a follow-up period of 9 years. Baseline serum or plasma samples were analysed for antibodies against human papillomavirus (HPV) types 6, 16 and 18 and Chlamydia trachomatis. In total, 114 cases (42 incident cases identified during follow-up and 72 prevalent cases identified at baseline) and 519 matched controls were included in the study. HPV-16 seropositivity was strongly associated with cervical cancer (OR=6.33; 95 % CI 3.45–11.62). Overall, C. trachomatis was not associated with cervical cancer, but was associated with cervical cancer in analyses restricted to incident cases of cancer (OR=2.94; 95 % CI 1.17–7.42) or to cases in which serum samples were analysed (OR=3.13; 95 % CI 1.16–8.47). An antagonistic interaction between HPV-6 and -16 was found in a multiplicative model. These results suggest that different HPV types might interfere in cervical carcinogenesis and that C. trachomatis is associated with cervical cancer in prospective studies, and support the notion that HPV-16 seropositivity is strongly associated with cervical cancer.

The risk for coronary events may rise during acute infection. Perturbation in coronary endothelial function emerges as one important link. We investigated whether simvastatin could protect the coronary arterial function from the adverse effects of acute infection in swine.Coronary endothelium-dependent and -independent vasomotor responses were assessed by Doppler velocimetry in 12 Chlamydia pneumoniae-infected and 6 sham-infected swine 2 weeks after intratracheal inoculation. Half of animals from the infection group were pre-treated with simvastatin (80 mg daily), while the remaining animals received placebo. The treatment was started 2 weeks prior to inoculation and continued until the end of the study. ANOVA was used for statistical calculations. Data are mean+/-S.D.All animals inoculated with C. pneumoniae developed IgM antibodies against this organism. As compared to noninfected animals, peak-to-baseline coronary flow velocity (CFV) ratio after bradykinin was significantly decreased in infected animals regardless of statin treatment (p=0.01). Intracoronary 10(-6) M acetylcholine caused slight dilatory responses in both noninfected and infected-treated animals (CFV ratio: 1.6+/-0.2 and 1.4+/-0.2, respectively; p0.1), while a velocity drop (CFV ratio: 0.7+/-0.1; p0.01 versus noninfected-infected and treated), indicating constriction, was observed in infected-nontreated animals; 10(-5) M acetylcholine caused vasoconstriction in all animals, with a significantly more prolonged response in the infected-nontreated group (p0.01). Intracoronary adenosine and SNP induced similar dilatory responses in all groups (p0.5). There were no differences in markers of systemic inflammation (fibrinogen, amyloid, and CRP) and lipid profile (HDL, LDL and total cholesterol) between the groups (p0.2).Acute infection is associated with impairment of the muscarinic and kinin-related reactivity of coronary circulation. These functional abnormalities are in part prevented by simvastatin through mechanisms unrelated to lipid lowering.

Introduction Chronic rejection (CHR) of organ allografts, one of the most significant problems in modern transplantation, is not fully understood. This study sought to evaluate the influence of selected parameters on late kidney transplant function. Patients and Method The studied group consisted of eighty-six patients who received allogeneic transplants between 1988 and 1999 for leukocyte Chlamydia pneumoniae–DNA, immunoglobulin (Ig)A/IgG anti–C pneumoniae, blood lipids, ischemic damage in the donor and during organ preservation, HLA mismatch, and acute rejection episodes. Results Eighty-six patients were segregated as 26 patients (30%) with histologically proven chronic graft rejection (CHR[+]) and 59 patients (70%) without (CHR[−]). The presence of C pneumoniae–DNA in peripheral blood leukocytes was significantly more frequent in CHR(+) than CHR(−) group (46% vs 20%). Patients with leukocytes positive for C pneumoniae–DNA more frequently (50%) had CHR than patients negative for C pneumoniae–DNA (22%). CHR(+) patients showed significantly lower HDL levels (47 mg/dL vs 58 mg/dL) and higher triglyceride levels (193 mg/dL vs 148 mg/dL). To study the cumulative effect of differences between the CHR(+) and CHR(−) groups, we applied a multiple binary logistic regression analysis. An econometric model enabled us to calculate the probability of CHR for a given patient taking into account covariates chosen by means of stepwise selection: the presence of C pneumoniae–DNA in blood leukocytes, the use of continuous pulsatile perfusion in hypothermia, myocardial infarction occurrence, and triglyceride concentrations. Conclusion The presence of C pneumoniae–DNA in peripheral blood leukocytes increased the risk of CHR, which may be predicted by a multifactor analysis of chosen parameters. (Less)

Background: Upregulation of proinflammatory endothelial cell adhesion molecules and decreased bioactivity of endothelial nitric oxide (NO) are important in the pathogenesis of atherosclerosis. We investigated the effects of co-infection with Chlamydia pneumoniae and Helicobacter pylori on these two events in apoE-KO mice. Methods: Thirty-two apoE-KO mice, 8 weeks old, were equally divided into 4 groups. The first 2 groups were infected with either C. pneumoniae or H. pylori, while the 3rd group was infected with both C. pneumoniae and H. pylori. Mice from the 4th group and 4 wild-type mice served as controls. Thoracic and abdominal aortas were harvested after 10 weeks, and staining for vascular cell adhesion molecule-1 (VCAM-1) and intracellular adhesion molecule-1 was analyzed by immunocytochemistry. The endothelial vasomotor responses of thoracic aortas to methacholine were studied in organ chambers in the absence and presence of L-NAME. The plasma levels of nitrate/nitrite were measured. Results: Staining for VCAM-1 was more intense at the branching sites of aortas from mice with co-infection than in mono-infected or noninfected apoE-KO mice. The relaxation responses to methacholine and the plasma levels of nitrate/nitrite were significantly less in the co-infected group than in the other groups (p < 0.05). Conclusion: Co-infection of apoE-KO mice with C. pneumoniae and H. pylori seems to be associated with impaired bioactivity of endothelial NO and increased expression of VCAM-1 at branching sites. The findings may suggest an additive interaction of these pathogens in atherogenesis.

A method for detection and genotyping of genital Chlamydia trachomatis infections based on omp1 gene amplification and sequencing was developed. DNA was extracted from urogenital or urine samples using a Chelex-based method, and an approximately 1,100-bp-long fragment from the omp1 gene was directly amplified and sequenced. Genotyping was performed by BLAST similarity search, and phylogenetic tree analysis was used to illustrate the evolutionary relationships between clinical isolates and reference strains. The method was used to determine the genotypes of C. trachomatis in 237 positive urogenital and/or urine specimens collected at a Swedish sexually transmitted disease clinic during 1 year. The most common genotypes corresponded to serotypes E (47%) and F (17%). The omp1 gene was highly conserved for genotype E (106 of 112 samples without any mutation) and F (41 of 42 samples without any mutation) strains but appear slightly less conserved for genotypes G ( n = 6) and H ( n = 6), where the sequences displayed one to four nucleotide substitutions relative to the reference sequence. Genotyping of samples collected at the follow-up visit indicated that two patients had become reinfected, while three other patients suffered treatment failure or reinfection. One woman appeared to have a mixed infection with two different C. trachomatis strains. This omp1 genotyping method had a high reproducibility and could be used for epidemiological characterization of sexually transmitted Chlamydia infections.

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.

Background —Arterial relaxation is largely regulated by endothelial nitric oxide (NO). Its diminished activity has been associated with incipient atherosclerosis. We investigated the endothelium-dependent relaxation of aorta in apolipoprotein E–knockout (apoE-KO) mice exposed to single or repeated Chlamydia pneumoniae inoculation. Methods and Results —Forty-eight apoE-KO mice, 8 weeks old, were inoculated intranasally with C pneumoniae (n=24) or saline (n=24) every 2 weeks over a 6-week period. Twenty mice (10 infected and 10 controls) were killed at 2 weeks and 6 weeks, respectively, after the first inoculation. The smooth muscle tone of aortic rings was measured in vitro at both time points. The norepinephrine-precontracted thoracic aortic rings were successively exposed to methacholine in the absence and presence of N G -nitro- l -arginine methyl ester (L-NAME) and diclofenac. The methacholine-induced relaxation was attenuated in the infected mice at 6 weeks in both the absence and presence of L-NAME ( P P P P Conclusions — C pneumoniae impairs arterial endothelial function, and the NO pathway is principally involved. Cyclooxygenase-dependent vasoconstricting products may also account for the infection-induced impaired relaxation. These findings further support the role of C pneumoniae infection in atherosclerosis development.

Background. To study the incidence of non-gonococcal salpingitis, gonococcal salpingitis and ectopic pregnancy in a defined population over a 28-year period on the assumption that the frequency of salpingitis and ectopic pregnancy may indirectly illustrate the epidemiological pattern of Chlamydia trachomatis. Design. A retrospective epidemiological study. Setting. University hospital with an urban catchment area. Patients. Five thousand two hundred and thirty-three patients admitted to the hospital between 1969 and 1996 with a diagnosis of ectopic pregnancy, non-gonococcal salpingitis, or gonococcal salpingitis. Results. The frequencies of both non-gonococcal and gonococcal salpingitis increased steeply early in the period under study, rising to a peak in the early 1970s, then decreasing throughout the period except for the last 3 years when a slight increase was seen again. The frequency of ectopic pregnancy showed a steady increase, peaking in the late 1980s and early 1990s and then declining at the end of the study period. While the introduction of more sensitive pregnancy tests and programs for assisted fertility would increase the rate of ectopic frequency the decline during the ʼnineties cannot be accounted for in this way. The peak of salpingitis cases in the early 'seventies seems to be mirrored exactly by the peak of ectopic pregnancies fifteen years later in the late 'eighties. Conclusion. The frequencies of salpingitis and of ectopic pregnancy can probably be used to estimate the incidence of preceding Chlamydia trachomatis. Thus the incidence of C. trachomatis has probably declined since the early 'seventies like that of N. gonorrheae.

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 °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.

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.

OBJECTIVE: To estimate the prevalence of Chlamydia trachomatis, C. psittaci and C. pneumoniae antibodies in sera from altogether 931 blood donors, patients with symptoms of urethritis, assumed salpingitis and sexually acquired reactive arthritis (SARA), and women with fertility problems. METHODS: IgG antibodies to C. trachomatis, C. psittaci and C. pneumoniae were determined by microimmunofluorescence (MIF) tests. All patients were also tested for genital C. trachomatis infection using direct immunofluorescence (DIF) tests. RESULTS: The DIF-positive cases had a significantly (p < 0.0001) higher prevalence of C. trachomatis antibodies than the DIF negatives, i.e. 88.5% versus 14% in men with urethritis, 94.3% versus 36.4% in women with salpingitis, 66.7% versus 16.7% in SARA patients and 90.6% versus 20.8% in women with fertility problems. Antibody reactivity to all three chlamydial species was found significantly (p < 0.0001) more often in the patient groups and in those with a DIF-confirmed genital C. trachomatis infection than in blood donors. CONCLUSIONS: Presence of serum antibodies to C. trachomatis is tightly associated with the presence of chlamydiae in the genital tract, which also influences the cross-reactivities occurring in the MIF tests between chlamydial species.

Objective The longitudinal epidemiological development of the new variant of Chlamydia trachomatis was studied after appropriate testing procedures had been introduced when the strain was detected in 2006. Methods The number of cases of the new variant of C trachomatis was followed from 2007 through 2011 from the laboratory records. Testing for C trachomatis is centralised to one laboratory with around 80–85 000 persons being tested annually in a population of 1.1 million. Results During the 5-year period, 410 973 patients were tested of which 25 723 cases were positive. The proportion of the new variant of all positive cases declined from 30% in 2007 to 6% in 2011. While the number of the new variant of C trachomatis declined, the ordinary wild-type strains remained largely unchanged. Conclusions A selective decline of the new variant of C trachomatis has occurred after appropriate laboratory testing was introduced. A new balance point between 5% and 10% for the new variant seems to be gradually approached.

Objective We sought to analyze the prevalence and clinical manifestations of Mycoplasma genitalium infection in a heterogeneous population of women. Study Design The study was designed as a cross-sectional case-control study. Women attending a gynecological outpatient service from 2003 through 2008 were invited to participate. Results The prevalence of M genitalium was 2.1% and of Chlamydia trachomatis was 2.8% among 5519 tested women. A total of 679 women were included. Both pelvic inflammatory disease (PID) and cervicitis were independently associated with M genitalium (odds ratio, 9.00; 95% confidence interval, 1.62–49.89 and odds ratio, 3.80; 95% confidence interval, 2.06–7.03, respectively). Women with C trachomatis had a higher frequency of both PID (18.3% vs 4.9%, P P M genitalium . Conclusion M genitalium was an independent and strong risk factor for both cervicitis and PID although, compared to C trachomatis , clinical manifestations were less frequent.

International audience; Chlamydia trachomatis is responsible for both trachoma and sexually transmitted infections, causing substantial morbidity and economic cost globally. Despite this, our knowledge of its population and evolutionary genetics is limited. Here we present a detailed phylogeny based on whole-genome sequencing of representative strains of C. trachomatis from both trachoma and lymphogranuloma venereum (LGV) biovars from temporally and geographically diverse sources. Our analysis shows that predicting phylogenetic structure using ompA, which is traditionally used to classify Chlamydia, is misleading because extensive recombination in this region masks any true relationships present. We show that in many instances, ompA is a chimera that can be exchanged in part or as a whole both within and between biovars. We also provide evidence for exchange of, and recombination within, the cryptic plasmid, which is another key diagnostic target. We used our phylogenetic framework to show how genetic exchange has manifested itself in ocular, urogenital and LGV C. trachomatis strains, including the epidemic LGV serotype L2b.

Background Disease severity in Mycoplasma pneumoniae (MP) infection could potentially be related to bacterial factors such as MP genotype (MP1 or MP2; distinguished by different adhesions proteins) or bacterial load in airway secretions. We have compared these parameters in patients who were hospitalized for MP pneumonia, with outpatients with mild MP disease. Methods MP bacterial load was measured by real-time PCR in 45 in- and outpatients ("clinical study group") in whom MP DNA had been detected in oropharyngeal secretions by PCR. In addition, genotype and phylogenetic relationships were determined. The phylogenetical assessment was done by partial DNA sequencing of the P1 gene on isolates from 33 patients in the clinical study-group where sufficient DNA was available. The assessment was further extended to isolates from 13 MP-positive family members and 37 unselected MP positive patients from the two subsequent years and two different geographical locations. In total 83 strains were molecular characterized. Results Mean MP loads were significantly higher in 24 hospitalized patients than in 21 outpatients (1600 vs. 170 genomic equivalents/μL, p = 0.009). This difference remained significant after adjustment for age and days between disease onset and sampling. Hospitalized patients also had higher C-reactive protein levels. Mean levels were 188 vs 20 mg/L (p = 0,001). The genotype assessment showed MP genotype 1 in 17 of the 33 sequenced strains from the clinical study-group, and type 2 in 16 of these patients. Within each genotype, sequence differences were minimal. No association between disease severity and MP genotype was observed. In the extended genotype assessment, MP1 was found in similar proportions. In family contacts it was found in 53% and among patients from the two subsequent years 53% and 40%. Conclusions A higher MP bacterial load in throat secretions at diagnosis was associated with more advanced respiratory disease in patients, but MP genotype did not influence disease severity. Both MP genotypes co-circulated during recent outbreaks in Sweden.

Background: Atherosclerosis is considered to be an inflammatory disease. Infections are a significant cause of inflammation. Acute infections might precipitate acute coronary syndromes (ACS) whereas chronic infections might be stimuli for the development of atherosclerosis. Methods: Coronary angiograms were done on 211 of 335 patients with ACS and the percentage of coronary obstruction was determined. Serum antibody levels to Chlamydia pneumoniae, C. pneumoniae heat shock protein 60 (CpnHSP60), human heat shock protein 60 (hHSP60), enterovirus (EV), herpes simplex virus (HSV), cytomegalovirus (CMV), and two major periodontal pathogens, Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis, were measured in healthy controls (n = 355) and all patients. Results: Serum antibody levels to periodontal pathogens did not correlate with ACS. However, IgA-class antibody levels to Aggregatibacter actinomycetemcomitans ( p = 0.021), CpnHSP60 ( p = 0.048) an hHSP60 ( p = 0.038) were higher in patients with coronary occlusion or obstruction compared to those without any obstruction. Odds ratios for coronary changes in the highest quartile as compared to the lower quartiles were for A. actinomycetemcomitans IgA 7.84 (95% CI 1.02—60.39, p = 0.048), for CpnHSP60 IgA 8.61 (1.12—65.89, p = 0.038), and for human HSP60 IgA 3.51 (0.79—15.69, p = 0.100). Conclusions: We have previously reported that EV and HSV titres correlated significantly to acute coronary events. They do not correlate to the degree of coronary obstruction as shown here. However, infection by A. actinomycetemcomitans or C. pneumoniae or host response against them associated with coronary obstruction. Clinical coronary events may arise by the effect of acute infections and obstructing lesions by a chronic inflammatory stimulus.

BACKGROUND:: In 2006, a new genetic variant of Chlamydia trachomatis (nvCT) was discovered in Sweden. Clinical manifestations of this infection were studied in a high-risk population. METHODS:: During 2007, a prospective case-control study on sexual lifestyle and urogenital infections was performed at the Centre for Sexual Health (CSH), affiliated to Malmo University Hospital. A total of 629 C. trachomatis positive cases and 1252 negative controls were included. At Malmo University Hospital, Department of Obstetrics and Gynecology, all cases of pelvic inflammatory disease (PID) were assessed and correlated to the prevalence of nvCT. RESULTS:: Patients with nvCT or wild type C. trachomatis (wtCT) infection did not differ regarding their sexual lifestyle. Men with nvCT or wtCT infection did not differ in uro-genital symptoms or clinical findings. Women with nvCT infection reported painful urination (12.2% vs. 25.8%, P = 0.02) and were diagnosed with urethritis (11.1% vs. 40.0%, P = 0.04) less often than women with wtCT infection. The ratio of lower abdominal pain in women with nvCT infection was only half of that in women with wtCT infection (13.4% vs. 27.8%, P = 0.02). PID was detected in 0.8% of women with C. trachomatis infection in Malmo. All these cases were due to wtCT infection. CONCLUSIONS:: Symptomatic urethral infection and lower abdominal pain was less common in women with nvCT as compared to wtCT. Infection with nvCT was more frequently asymptomatic suggesting a possible difference in virulence between the nvCT strain and the wtCT strain. (Less)

BACKGROUND: Infections caused by Chlamydia pneumoniae are considered to participate in inflammatory processes leading to coronary artery disease. After a primary infection, the bacteria remain dormant intracellularly causing a chronic inflammatory stimulus. MATERIALS AND METHODS: Blood samples were obtained from 235 patients with acute myocardial infarction (AMI) and 108 patients with unstable angina pectoris (UA). We evaluated the prognostic significance of bacterial and viral antibody titers, serum troponin T, C-reactive protein, and chlamydial lipopolysaccharide (cLPS) concentrations during acute coronary syndrome of patients with AMI and UA for cardiovascular death and new UA and AMI that required hospital care during a 6-year follow-up. RESULTS: Serum cLPS levels correlated with C-reactive protein and serum troponin T concentrations during acute coronary events. Patients with AMI had significantly higher serum concentration of cLPS compared with patients with UA. Enterovirus antibody titers and cholesterol-lowering therapy at admission of the index event were negatively correlated with cLPS concentration (r = -.198, P = .0003 and r = -.26, P = .019, respectively). The presence of circulating cLPS was associated with a hazard ratio of 2.04 for a new cardiovascular event during the follow-up period (P = .006). The area under the curve in the receiver operating graph was .572. CONCLUSION: cLPS is evidently liberated from the infected atherosclerotic tissue during an acute coronary event. Our study supports the view that inflammation caused by C. pneumoniae infection is an important but as yet poorly understood factor in the development of atherosclerosis and may play a role in acute vascular events. (Less)

The human polymnaviruses JC virus (JCV) and BK virus (BKV) are oncogenic in experimental systems and commonly infect humans. JCV DNA has been reported to be present in human colon mucosa and in colorectal cancers. To investigate whether the risk for colorectal cancer is associated with JCV or BKV infection, we performed a case-control study nested in the Janus biobank, a cohort of 330,000 healthy Norwegian subjects. A 30-year prospective follow-up using registry linkages identified 386 men with colorectal cancer who had baseline serum samples taken >3 months before diagnosis. Control subjects were matched for sex, age and date of blood sampling and county of residence. Seropositivity for JCV or BKV had high (97-100%) sensitivity for detection of viral DNA-positive subjects and discriminated the different polyomaviruses. Seropositivity was mostly stable over time in serial samples. The relative risk for colorectal cancer among JCV seropositive subjects was 0.9 (95% CI: 0.7-1.3) and the BKV-associated relative risk was 1.1 (95% CI: 0.8-1.5). Determining seropositivity using alternative cutoffs also found no evidence of excess risk. In summary, this prospective study found no association between JCV or BKV infections and excess risk for colorectal cancer.

Aims: The aim of the study was to explore whether exposure to microbial agents determines the prevalence of acute coronary events. Methods and results: Patients with unstable angina pectoris and myocardial infarction (N = 335) and their paired controls were investigated. The subjects answered a questionnaire about their childhood contagious diseases: varicella, scarlet fever, measles, rubella, mononucleosis and mumps. Blood samples were taken for bacterial and viral serology. The odds ratio for CHD was highest in the upper quartile of the enterovirus (EV), herpes simplex virus (HSV) and Chlamydia pneumoniae HSP60 IgG antibody titers (1.86, p = 0.001, 1.57, p < 0.048 and 1.70, p = 0.016, respectively). The antibody titers increased cumulatively the risk for CHD (odds ratios 1.89, 2.24, 3.92 and p-values < 0.001, 0.001 and 0.047). Childhood contagious diseases (n = 6) had a protecting effect against CHD (odds ratio 0.86, p = 0.013). The risk for acute coronary events decreased significantly with increasing number of childhood contagious diseases (p = 0.007). Conclusions: Infections have a dual role in the genesis of CHD. EV, HSV and C. pneumoniae heat shock protein 60 IgG antibodies are associated with increased risk for CHID. Protection from infections usually suffered during the childhood before the era of MMR vaccination may predispose the individual to CHD. (Less)

Human papillomavirus (HPV) persistence is the major cause of cervical cancer, but most HPV infections will not persist and risk factors for HPV persistence are not well known. Chlamydia (C.) trachomatis infection seems to also be associated with cervical cancer. We investigated whether C. trachomatis infection is a risk factor for HPV persistence. In a cohort of 12,527 women participating in a population-based HPV screening trial in Sweden, 6,418 women completed testing for HPV DNA by general primer PCR and typing by reverse dot blot hybridization. On average 19 months later, 303 women that had been HPV-positive and had normal cytology at enrollment completed a new HPV test. Environmental exposures were assessed by an 87-item questionnaire. Previous sexually transmitted infections were also investigated by serology. At follow-up, 44% of the women were positive for the same type of HPV DNA as at enrollment. Persistence correlated with length of follow-up (p < 0.01) and condom use seemed to protect against HPV persistence (p < 0.05). The most significant risk factor for persistent presence of HPV DNA was self-reported history of previous C. trachomatis infection (relative risk in multivariate model = 2.09; 95% confidence interval = 1.05-4.18). We conclude that persistence of oncogenic HPV infections is more likely among women with a previous C. trachomatis infection.

Aim: The purpose of the study was to assess the association between seropositivity to various infectious agents and stable coronary artery disease (CAD), controlling simultaneously for a variety of potential confounders. We also investigated whether the choice of a larger reference group might affect the results, and whether or not seropositivity to multiple agents was associated with a systemic inflammatory response. Methods: We assessed the simultaneous prevalence of antibodies against Helicobacter pylori, Chlamydia, cytomegalovirus, and herpes simplex virus in 312 patients with angiographically proven coronary artery disease (CAD) and in 479 age and sex matched controls. C-reactive protein, interleukin-6, fibrinogen, PAI- I -activity, D-dimer, von Willebrand Factor, plasma viscosity, and a complete blood cell count were determined in all subjects. Results: Seropositivity to all of the four agents was 21.8% in cases and 13.6% in controls (P = 0.0003). We found a dose-response relationship between combined IgG-seropositivity to H. pylori, Chlamydia, cytomegalovirus, and herpes simplex virus and odds for the presence of angiographically confirmed stable CAD which, however, was strongly reduced after controlling for a variety of potential confounders. The dose-response pattern was no longer evident if a more stable reference group (subjects seropositive for two agents) was used instead of the relatively small reference group with zero or one seropositivity. We found no consistent pattern between IgG-seropositivity to several pathogens and inflammatory markers. Conclusions: Based on serological evidence of various infectious agents, this study suggests that the aggregate number of persistent infections is not independently associated with an increased risk for CAD if control for confounding and use of a stable reference group are guaranteed. (C) 2003 Published by Elsevier Ireland Ltd. (Less)

Background: Coronary endothelial dysfunction contributes to the pathogenesis of acute coronary syndromes (ACSs). Acute Chlamydia pneumoniae infection has been epidemiologically associated with ACS. In this study, we investigated whether acute C. pneumoniae infection could alter the endothelial vasomotor function of porcine coronary vessels. Methods and results: Twenty pigs, 7–9 kg in weight, were inoculated intratracheally with C. pneumoniae (n=12) or saline (n=8), and investigated at 3 days (five infected/four non-infected) and 2 weeks (5+2 infected/four non-infected) after inoculation. The endothelium-dependent reactivity of coronary microcirculation was assessed at both time points by measuring peak coronary flow velocity (CFV) in response to bradykinin, before and after infusions with glutathione, an antioxidant, and Image-arginine, a substrate for nitric oxide synthase (NOS). CFV after bradykinin was significantly decreased in infected animals at both time points. At 2 weeks, both glutathione and Image-arginine significantly improved CFV after bradykinin. CFV after sodium nitroprusside (SNP) was similar in both groups. At 3 days, the relaxation responses of bradykinin-induced pre-contracted left anterior descending (LAD) coronary rings to bradykinin were significantly less in infected animals. NG-nitro-Image-arginine-methyl-ester, an NOS inhibitor, had significantly greater inhibitory effect on bradykinin-induced relaxation in infected animals. Plasma nitrate–nitrite and fibrinogen, and NOS activity from LAD coronary samples were significantly increased in infected animals. Conclusion: Acute C. pneumoniae infection causes endothelial dysfunction of both resistance and epicardial coronary vessels, and favours a pro-coagulant status. These effects could in part account for the epidemiologically suggested association between acute infection and ACS. (Less)

Our objectives were to compare the prevalence of sexually transmitted infections (STIs) in 103 women undergoing induced legal abortion (LA) and in 101 women with confirmed, recent illegal abortion (IA), in Maputo, Mozambique. For the purpose of this study, LA was considered the abortion provided in the Maputo Central Hospital with the approval of the Ministry of Health, and IA the one not provided through the approved facility, mentioned above. Women with IA were recruited in the outpatient gynaecology ward and women with LA in the emergency gynaecology ward in the Maputo Central Hospital, during the same time period. Serological tests for syphilis (rapid plasma reagin, ELISA-IgG and fluorescent treponemal antibody absorption), gonorrhoea (indirect haemagglutination) and chlamydia (microimmunofluorescence) were carried out. Direct immunofluorescence for detection of Chlamydia trachomatis antigen was done on endocervical smears from all the women. The prevalence of syphilis seropositivity in IA women is twice that of LA women (odds ratio [OR] 2.13; 95% confidence interval [CI] 0.55-9.95), with 10.9% and 4.9%, respectively. Exposure to gonorrhoea in these 2 groups is similar (OR 1.18; 95% CI 0.63-2.20), with seroprevalence of 31.1% in the LA and 34.7% in the IA group. The high titres are also similar in both groups. Serology findings for C. trachomatis indicate prevalence of seropositivity of 40.6% in the LA and 44.4% in the IA group with no significant difference (OR 1.17; 95% CI 0.64-2.13). In conclusion, STIs are highly prevalent in both IA and LA groups in Maputo. Urgent interventions are needed to reduce their prevalence and consequently their adverse consequences.

The reported rate of detection of Chlamydia pneumoniae DNA within atherosclerotic lesions by PCR varies between 0 and 100%. In this study, identical sets of coded experimental atheroma samples ( n = 15) and spiked controls ( n = 5) were analyzed by 16 test methods in nine centers by means of PCR. The positive controls were correctly identified to levels of 1, 0.1, and 0.01 inclusion bodies of C. pneumoniae /ml of tissue homogenate by 16 (100%), 11 (69%), and 3 (19%) of the test methods, respectively. Three out of 16 negative controls (19%) were rated positive. Positivity rates for atheroma samples varied between 0 and 60% for the different test methods, with the maximum concordant result for positivity being only 25% for one carotid artery sample. There was no consistent pattern of positive results among the various laboratories, and there was no correlation between the detection rates and the sensitivity of the assay used.

The lack of standardization in chlamydia serology has made interpretation of published data difficult. This study was initiated to determine the extent of interlaboratory variation of microimmunofluorescence (MIF) test results for the serodiagnosis of Chlamydia pneumoniae infections. Identical panels of 22 sera were sent to 14 laboratories in eight countries for the determination of IgG and IgM antibodies by MIF. Although there was extensive variation in the numeric titer values, the overall percentage agreement with the reference standard titers from the University of Washington was 80%. For results by serodiagnostic category, the best agreement was for four-fold rise in IgG titers, while the lowest agreement was for negative or low IgG titers. Agreement for IgM titers was 50%-95%. Four laboratories failed to discern false-positive IgM titers possibly because of the presence of rheumatoid factor. Further studies are underway to determine the source of interlaboratory variation for the MIF test.

A commercial test (rELISA) based on a recombinant chlamydial lipopolysaccharide (LPS) antigen has been evaluated for the diagnosis of acute infections caused by Chlamydia pneumoniae (TWAR) and Chlamydia psittaci. This test and a microimmunofluorescence test (MIF) were compared in 160 patients with community-acquired pneumonia. Seventeen of nineteen cases with significant titre changes detected by rELISA were confirmed by MIF. The two remaining cases not confirmed by MIF were considered false-positive reactions. One case positive by MIF only was judged not to be a true-positive reaction. All three cases occurred in patients with Mycoplasma pneumoniae infection and may be the result of a mitogenic effect. High antibody titres have been used to indicate acute C. pneumoniae infection. We found high MIF or rELISA titres to be equally common in patients and controls; no association between the two tests was detected. An unexpected cross-reactivity between the rELISA antigen and parvovirus was observed, which might have diagnostic implications. Both MIF and rELISA detected acute C. pneumoniae and C. psittaci infection, and there was good agreement between the tests. Single serum diagnosis was generally not feasible with either MIF or rELISA.

The antibody response to heat shock proteins 60 and 10 were studied in 163 patients with tubal factor infertility and in 163 age-matched pregnant women. The associations of these antibodies with specific antibodies to Chlamydia trachomatis and to Chlamydia pneumoniae as well as with antibodies to the common chlamydial lipopolysaccharide antigen were studied. Patients with tubal factor infertility had significantly higher frequencies and titres of all antibodies except to C. pneumoniae. In a logistic regression model an association was found between the prevalence of antibodies to the heat shock proteins and to C. trachomatis but no independent influence of antibodies to C. pneumoniae. No interaction between C. trachomatis and C. pneumoniae suggesting a synergistic effect was found although the heat shock proteins from these two organisms are immunologically similar. Antibodies to the chlamydial lipopolysaccharide also seemed to be related to C. trachomatis and not to C. pneumoniae in these women.

Nested polymerase chain reaction (nPCR) demonstrated the presence of Chlamydia pneumoniae-specific DNA in peripheral blood mononuclear cells (PBMC). PBMC samples were obtained from 103 consecutive patients (62 male, 41 female) aged 22-85 years (mean, 64) admitted for coronary angiography because of suspected coronary heart disease and from 52 blood donors (43 male, 9 female) aged 40-64 years (mean, 49). Of the 101 evaluable patients, 60 (59%) were identified by nPCR assay as C. pneumoniae DNA carriers; C. pneumoniae-specific microimmunofluorescence (MIF) serology confirmed exposure to the bacterium in 57 (95%) of the 60 nPCR-positive patients. Among the 52 blood donors, the nPCR assay identified 24 (46%) C. pneumoniae DNA carriers, all of whom were positive by C. pneumoniae-specific serology. Thirty-two patients (32%) and 23 blood donors (44%) were MIF antibody-positive but repeatedly nPCR-negative; Bartonella henselae- or Bartonella quintana-specific antibodies were not detected among any of these subjects. In this study, C. pneumoniae DNA was common in PBMC of patients with coronary heart disease and in middle-aged blood donors.

Background Genital chlamydia infections often are asymptomatic, which promotes their spread in the population. In women, the possible consequences of infection are pelvic inflammatory disease and infertility. Most studies on the prevalence of Chlamydia trachomatis have been based on clinical series, and prevalences tend to vary with the clinical setting. Few seroepidemiologic studies have emerged from industrialized countries. Goal of this Study To assess the prevalence of Chlamydia trachomatis using culture and serology, and its relationship with possible risk factors. Study Design This was a population-based study involving completion of a self-administered questionnaire, analysis of cervical samples for Chlamydia trachomatis, and serologic tests for Chlamydia trachomatis antibodies. Results The prevalence of Chlamydia trachomatis infection was 2.7 %, and the seroprevalence was 24.7 % among the sexually active women. Seropositivity was correlated with sexual behavior variables, and the incidence of serologic cross-reactivity with respiratory infections (strain TWAR) was low. Multivariate logistic regression analysis showed the number of sexual partners, age at first coitus, history of therapeutic abortion, and previous pelvic inflammatory disease to be independently correlated with seropositivity. Conclusion Early sexual experience and multiple lifetime sexual partners are independent risk factors for Chlamydia trachomatis infection.

Background: Serology is often used for the diagnosis of Mycoplasma pneumoniae. It is important to identify specific antigens that can distinguish between the presence or absence of antibodies against M. pneumoniae. The two proteins, P116 and P1, are found to be immunogenic. By using these in ELISA it is possible to identify an immune response against M. pneumoniae in serum samples. Results: A recombinant protein derived from the P116 protein and one from the P1 protein were used in two ELISA tests, rP116-ELISA and rP1-ELISA. Human serum samples from patients with atypical pneumonia were tested and compared to the results of the complement fixation test. There was a good agreement between the two tests but the rP1-ELISA showed the best discrimination between positive and negative samples. Conclusion: Two ELISA tests based on recombinant proteins have been analysed and compared to the complement fixation test results. The two ELISA tests were found suitable for use in serodiagnostics of M. pneumoniae infections. The use of specific antigens eliminates the risk of cross reaction to an immune response against other bacteria.

Serum antibodies to Chlamydia trachomatis were studied by microimmunofluorescence (micro-IF) testing and by immunoblotting among 52 women with C. trachomatis cervical infection. All women underwent therapeutic abortion, and 10 (19.2%) subsequently developed laparoscopically confirmed salpingitis. Women who developed salpingitis had lower geometric mean titers of micro-IF antibody before abortion (14.9 x/divided by 2.3) than did women who did not develop salpingitis (41.6 x/divided by 4.9, P less than .01). Women who developed salpingitis significantly less often had serum IgA antibodies to a 60-kilodalton (kDa) chlamydial antigen (P = .02) and IgG antibodies to antigens of 75-kDa (P = .008), 60-kDa (P = .03), and 57-kDa (P = .0003). Serum antibodies to 100-kDa, 32-kDa, and 29-kDa antigens occurred only in women who did not develop salpingitis. Differences in antibody prevalence to specific chlamydial antigens were not due to differences in serum antibody titers between the two groups. No correlation between neutralizing sera and the risk of postabortal salpingitis was detected.

Background: The etiology and significance of flu-like symptoms often appearing before myocardial infarction should be clarified. Methods: In a case-control study of 323 matched controls and a random sample of 110 out of 351 cases the presence of infection symptoms during the preceding four weeks before admission were asked and blood samples taken. Results: Enterovirus (EV), herpes simplex virus (HSV), and Chlamydia pneumoniae IgA titers were significantly higher in cases than in controls ( p Conclusions: Odds ratios for a coronary event in a logistic regression model were 4.79 for fatigue and 2.72 for EV antibody levels in their fourth quartile. A linear-by-linear association test showed increasing number of single symptoms with higher EV titer quartiles ( p = 0.004).

The humoral immune response to Chlamydia outer membrane protein 2 (Omp2) was studied. Omp2 is a highly genus-conserved structural protein of all Chlamydia species, containing a variable N-terminal fragment. To analyze where the immunogenic parts were localized, seven highly purified truncated fusion proteins constituting different regions of the protein were produced ( Chlamydia pneumoniae -Omp2 aa23-aa93 , Chlamydia psittaci -Omp2 aa23-aa94 , and Chlamydia trachomatis -Omp2 aa23-aa84, aa87-aa547, aa23-aa182, aa167-aa434, aa420-aa547 ). By an enzyme-linked immunosorbent assay with serologically defined patient sera, Omp2 was found to be a major immunogen of both C. pneumoniae and C. trachomatis infections ( P ≪ 0.0001). The humoral immune responses were not confined to any particular region of the Omp2 protein, and no species-specific anti-Omp2 immunoglobulins were detected.

Background Chlamydia trachomatis is the most common cause of sexually transmitted infections globally and the leading cause of preventable blindness in the developing world. There are two biovariants of C. trachomatis: 'trachoma', causing ocular and genital tract infections, and the invasive 'lymphogranuloma venereum' strains. Recently, a new variant of the genital tract C. trachomatis emerged in Sweden. This variant escaped routine diagnostic tests because it carries a plasmid with a deletion. Failure to detect this strain has meant it has spread rapidly across the country provoking a worldwide alert. In addition to being a key diagnostic target, the plasmid has been linked to chlamydial virulence. Analysis of chlamydial plasmids and their cognate chromosomes was undertaken to provide insights into the evolutionary relationship between chromosome and plasmid. This is essential knowledge if the plasmid is to be continued to be relied on as a key diagnostic marker, and for an understanding of the evolution of Chlamydia trachomatis. Results The genomes of two new C. trachomatis strains were sequenced, together with plasmids from six C. trachomatis isolates, including the new variant strain from Sweden. The plasmid from the new Swedish variant has a 377 bp deletion in the first predicted coding sequence, abolishing the site used for PCR detection, resulting in negative diagnosis. In addition, the variant plasmid has a 44 bp duplication downstream of the deletion. The region containing the second predicted coding sequence is the most highly conserved region of the plasmids investigated. Phylogenetic analysis of the plasmids and chromosomes are fully congruent. Moreover this analysis also shows that ocular and genital strains diverged from a common C. trachomatis progenitor. Conclusion The evolutionary pathways of the chlamydial genome and plasmid imply that inheritance of the plasmid is tightly linked with its cognate chromosome. These data suggest that the plasmid is not a highly mobile genetic element and does not transfer readily between isolates. Comparative analysis of the plasmid sequences has revealed the most conserved regions that should be used to design future plasmid based nucleic acid amplification tests, to avoid diagnostic failures.