Helicobacter pylori infection combined with non-alcoholic fatty liver disease increase the risk of atherosclerosis: Focus in carotid artery plaque.

Atherosclerosis has severe consequences on human health. Carotid artery plaques are a condition typically caused by atherosclerosis. Previous studies showed that nonalcoholic fatty liver disease (NAFLD) and Helicobacter pylori (H pylori) are risks factors for carotid artery plaque formation. We hypothesize that the combination of NAFLD with H pylori infection increases the risk of carotid artery plaque formation.A total of 4669 subjects aged > 40 years who underwent routine health checkups between January 2006 and December 2015 were retrospectively reviewed. A serial examination, including abdominal ultrasound, carotid artery ultrasound and esophago-gastroduodenoscopy (EGD), and biopsy urease testing, was conducted.In total, 2402 subjects were enrolled. There were no differences in H pylori infection status among patients with or without NAFLD. There was a trend of more participants with both NAFLD and H pylori infection (number [N]=583) presenting carotid artery plaque (N = 187,32.08%) than participants without NAFLD and H pylori infection (N = 589) who presented plaque formation (N = 106, 18.00%). Participants who had both H pylori infection and NAFLD had the highest risk of any carotid artery plaque (odds ratio [OR], 1.93; 95% confidence interval [CI], 1.413-2.636) based on a multivariate logistic regression analysis. This analysis also showed that age >60 years, male sex, low-density lipoprotein (LDL) >130 mg/dL, and H pylori infection were independent risk factors for concomitant NAFLD and carotid artery plaque formation.The combination of H pylori infection and NAFLD increases carotid artery plaque formation. H pylori eradication and NAFLD control may be warranted to prevent carotid artery plaque formation.

Plasma microbiome-modulated indole- and phenyl-derived metabolites associate with advanced atherosclerosis and postoperative outcomes.

OBJECTIVE: Multiple studies have shown that gut microbes contribute to atherosclerosis, and there is mounting evidence that microbial metabolism of dietary nutrients influences pathophysiology. We hypothesized that indole- and phenyl-derived metabolites that originate solely or in part from bacterial sources would differ between patients with advanced atherosclerosis and age- and sex-matched controls without clinically apparent atherosclerosis. METHODS: Plasma from the advanced atherosclerosis cohort (n = 100) was from patients who underwent carotid endarterectomy, open infrainguinal leg revascularization, or major leg amputation for critical limb ischemia. The controls (n = 22) were age- and sex-matched participants who had no peripheral arterial disease or history of stroke or myocardial infarction. Patients with chronic kidney disease were excluded. Metabolites and internal standards were measured using high-performance liquid chromatography and tandem mass spectrometry. RESULTS: Plasma metabolite concentrations differed significantly between the advanced atherosclerosis and control cohorts. After adjustment for traditional atherosclerosis risk factors, indole (odds ratio [OR], 0.84; 95% confidence interval [CI], 0.75-0.95; P = .004), tryptophan (OR, <0.001; 95% CI, <0.001-0.003; P < .001), indole-3-propionic acid (OR, 0.27; 95% CI, 0.019-0.91; P = .02), and indole-3-aldehyde (OR, 0.12; 95% CI, 0.014-0.92; P = .04) concentrations negatively associated with advanced atherosclerosis, whereas the kynurenine/tryptophan ratio (OR, 61.7; 95% CI, 1.9->999; P = .02) was positively associated. Furthermore, tryptophan and indole-3-propionic acid concentrations (Spearman coefficients of 0.63 and 0.56, respectively; P < .001) correlated with the ankle-brachial index, a surrogate for overall atherosclerotic disease burden. Fourteen patients experienced a major postoperative cardiac complication within 30 days in the advanced atherosclerosis cohort, which was associated with baseline kynurenine/tryptophan ratio (P = .001) and hippuric acid (P = .03). In a multivariate analysis, only the kynurenine/tryptophan ratio remained significantly associated with a postoperative cardiac complication (OR, 44.1; 95% CI, 3.3-587.1; P = .004). Twenty patients in the advanced atherosclerosis cohort experienced a major adverse cardiac event during the follow-up period, which was associated with hippuric acid (P = .002) and the kynurenine/tryptophan ratio (P < .001) at baseline. Both hippuric acid and the kynurenine/tryptophan ratio were independently associated with a major adverse cardiac event in multivariate analyses that included diabetes mellitus. CONCLUSIONS: Specific microbe-derived metabolite signatures associate with advanced human atherosclerosis and postoperative cardiac complications. We suggest that these metabolites are potential novel biomarkers for atherosclerotic disease burden and that further investigation into mechanistic links between defined microbial metabolic pathways and cardiovascular disease is warranted.

Bacterial profile in human atherosclerotic plaques.

BACKGROUND AND AIMS: Several studies have confirmed the presence of bacterial DNA in atherosclerotic plaques, but its contribution to plaque stability and vulnerability is unclear. In this study, we investigated whether the bacterial plaque-profile differed between patients that were asymptomatic or symptomatic and whether there were local differences in the microbial composition within the plaque. METHODS: Plaques were removed by endarterectomy from asymptomatic and symptomatic patients and divided into three different regions known to show different histological vulnerability: A, upstream of the maximum stenosis; B, site for maximum stenosis; C, downstream of the maximum stenosis. Bacterial DNA composition in the plaques was determined by performing 454 pyrosequencing of the 16S rRNA genes, and total bacterial load was determined by qPCR. RESULTS: We confirmed the presence of bacterial DNA in the atherosclerotic plaque by qPCR analysis of the 16S rRNA gene but observed no difference (n.s.) in the amount between either asymptomatic and symptomatic patients or different plaque regions A, B and C. Unweighted UniFrac distance metric analysis revealed no distinct clustering of samples by patient group or plaque region. Operational taxonomic units (OTUs) from 5 different phyla were identified, with the majority of the OTUs belonging to Proteobacteria (48.3%) and Actinobacteria (40.2%). There was no difference between asymptomatic and symptomatic patients, or plaque regions, when analyzing the origin of DNA at phylum, family or OTU level (n.s.). CONCLUSIONS: There were no major differences in bacterial DNA amount or microbial composition between plaques from asymptomatic and symptomatic patients or between different plaque regions, suggesting that other factors are more important in determining plaque vulnerability.

Lemierre's Syndrome Associated Mycotic Aneurysm of the External Carotid Artery with Primary Internal Carotid Artery Occlusion in a Previously Healthy 18-Year-Old Female.

Lemierre's syndrome is a rare life-threatening condition characterized by internal jugular vein thrombosis and is typically associated with a gram-negative infection with septic metastasis secondary to a retropharyngeal abscess that involves the vasculature of the head and neck. We report a case of Lemierre's syndrome in an 18-year-old female adolescent who developed an internal carotid artery occlusion and ipsilateral external carotid artery (ECA) mycotic aneurysm complicated by fulminant pseudomonal sepsis. The patient was managed with open ligation of the ECA with essentially complete recovery.

[The relationship between inflammatory periodontal diseases and cardiovascular diseases].

Estimated the relationship between inflammatory periodontal diseases and the risk of developing cardiovascular disease. Installed combined effect of periodontitis and cardiovascular disease.

The presence of some cytokines and Chlamydia pneumoniae in the atherosclerotic carotid plaque in patients with carotid artery stenosis.

BACKGROUND: Over the last few years the role of microorganisms in the pathogenesis of atherosclerosis has been widely discussed. Chlamydia pneumoniae activates immune cells to produce cytokines that are responsible for the formation of atheromatous carotid lesions. MATERIAL AND METHODS: The study was carried out at the Department of Vascular, General and Transplantation Surgery, Wroclaw Medical University, in 2002-2003, on 100 consecutive symptomatic patients with internal carotid stenosis, who underwent an endarterectomy procedure. Each patient had their carotid artery sampled in order to find C. pneumoniae DNA using the nested PCR method and some cytokines (TGF-ß, VEGF, FGF, TNF-α) using immunohistochemical examination. The control group consisted of 20 young organ donors who had been diagnosed with brain death and who had their healthy carotid artery harvested. Analogous genetic and immunohistochemical tests were performed. RESULTS: We did not confirm the presence of either cytokines or C. pneumoniae in the healthy carotid arteries. The presence of FGF was probably due to intima fibroblast activity, which is responsible for elastin and collagen synthesis for the extracellular matrix. C. pneumoniae was discovered in 68% of patients with carotid plaques. Three cytokines (TGF-ß, FGF, TNF-α) were detected in atherosclerotic internal carotid arteries as well. CONCLUSION: Chronic infection by C. pneumoniae may exacerbate carotid plaque development and may lead to its destabilization.

Severe narrowing of left cavernous carotid artery associated with Fusobacterium necrophorum infection.

Severe oropharyngeal infection can result in Lemierre's disease, a syndrome with high mortality secondary to inflammation and thrombosis of cervical and intracranial veins with involvement of contiguous structures; however arterial involvement is rare. We report a case of Lemierre's disease in a 12 year old boy with severe narrowing of the left cavernous carotid artery.

Prevotella nigrescens and Porphyromonas gingivalis are associated with signs of carotid atherosclerosis in subjects with and without periodontitis.

BACKGROUND AND OBJECTIVE: Oral microorganisms may be involved in the development of cardiovascular diseases, and Porphyromonas gingivalis is one of the periodontal microorganisms that has been found in carotid atheroma. The aim of this work was to study subgingival microorganisms and early carotid lesions in subjects with and without periodontitis. MATERIAL AND METHODS: Eighty-eight subjects with periodontitis and 40 subjects without periodontitis underwent dental examinations in 2003. The presence of the periodontal microorganisms Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens and Tannerella forsythia was analyzed from subgingival plaque using PCR amplification. The common carotid artery was scanned using ultrasound and the calculated intima-media area (cIMA) was measured. The association between periodontitis, the cIMA value and the presence of periodontal microorganisms, together with several confounders, was studied in a multiple logistic regression model. RESULTS: Smoking [odds ratio (OR) = 5.64; p = 0.001), level of education (OR = 5.02; p < 0.05) and the presence of P. gingivalis (OR = 6.50; p < 0.05) were associated with periodontitis. Explanatory factors for the increased cIMA were periodontitis (OR = 4.22; p < 0.05), hypertension (OR = 4.81; p < 0.05), high body mass index (OR = 5.78; p < 0.01), male gender (OR = 3.30; p < 0.05) and poor socioeconomic status (OR = 4.34; p < 0.05). P. nigrescens (OR 4.08; p < 0.05) and P. gingivalis (OR 7.63; p < 0.01) also appeared as explanatory variables associated with increased cIMA values. CONCLUSION: This cross-sectional study showed that P. nigrescens and P. gingivalis were significantly associated with increased cIMA values.

Detection of periodontal bacteria in atheromatous plaque by nested polymerase chain reaction.

BACKGROUND: In recent years, increasing evidence regarding the potential association between periodontal diseases and cardiovascular diseases has been identified. The available evidence underlines the importance of detecting periodontal pathogens on atheromatous plaque as the first step in demonstrating the causal relationship between the two conditions. The main aim of this investigation is to detect periodontitis-associated bacteria from carotid artery atheromatous plaque from patients who received an endarterectomy using strict sample procurement and laboratory procedures. METHODS: Atheromatous plaque from endarterectomies from carotid arteries were scraped and homogenized, and bacterial DNA was extracted. To obtain a representative concentration of amplicons, two amplifications of the bacterial 16S ribosomal-RNA gene were carried out for each sample with universal eubacteria primers by a polymerase chain reaction (PCR). A nested PCR with specific primers for the target bacteria was performed next. Statistical tests included the χ(2) test. RESULTS: Forty-two atheromatous plaque were analyzed. All of them were positive for ≥1 target bacterial species. The bacterial species most commonly found was Porphyromonas gingivalis (78.57%; 33 of 42), followed by Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans) (66.67%; 28 of 42), Tannerella forsythia (previously T. forsythensis) (61.90%; 26 of 42), Eikenella corrodens (54.76%; 23 of 42), Fusobacterium nucleatum (50.00%; 21 of 42), and Campylobacter rectus (9.52%; four of 42). The simultaneous presence of various bacterial species within the same specimen was a common observation. CONCLUSION: Within the limitations of this study, the presence of DNA from periodontitis-associated bacteria in carotid artery atheromatous plaque retrieved by endarterectomy is confirmed.

The role of infection in carotid plaque pathogenesis and stability: the clinical evidence.

INTRODUCTION: Chlamydia pneumoniae was the first pathogen linked with carotid atherosclerotic changes and plaque rupture. Currently, other common pathogens are also under investigation as potential contributors. METHODS: A systematic review of PubMed and Scopus databases was performed. Studies evaluating the infectious burden between symptomatic and asymptomatic patients with carotid plaque disease (CPD) were included. Furthermore, trials referring to common infectious agents (other than C. pneumoniae) incriminated for contribution in CPD were analyzed separately. RESULTS: Forty four studies were identified; 6 investigated the connection of infection with the plaque destabilization, 3 of which reported a significant association between infection and symptoms. Studies retrieved for the investigation of agents other than C. pneumoniae were: 18 about viruses, 16 about other bacteria and 4 examining both. Significant association or high detection rates of agents' genome or specific antibodies with CPD characteristics (intima media thickness values > 1 mm or symptoms) were found in a number of studies: 3 for HCV, 2 for CMV and 1 for enterovirus, EBV, HBV, and HIV. Moreover 4 studies about dental pathogens (i.e. Porpyromonas gingivalis), 5 about H. Pylori strains and 1 about Borrelia burgdorferi were identified supporting a positive association. CONCLUSION: There is considerable evidence supporting the contribution of other commonly encountered pathogens in the pathogenesis and rupture of the carotid plaque. Research in this direction should not be abandoned and further studies are necessary to elucidate the exact role of common infections in the pathogenesis and development of CPD and how this can be translated into novel pharmacological approaches for prevention and treatment.

Chlamydial lipopolysaccharide (cLPS) is present in atherosclerotic and aneurysmal arterial wall--cLPS levels depend on disease manifestation.

BACKGROUND: The role of Chlamydia pneumoniae in peripheral atherosclerosis disease and abdominal aortic aneurysm (AAA) remains unclear. Chlamydial lipopolysaccharide (cLPS) detection is a method used conventionally in routine chlamydial diagnosis of gynecological or ophthalmic samples. METHODS: We compared cLPS concentrations, as well as other markers of bacterial load, in plaques and sera of patients operated on for carotid artery stenosis (n=110), aorto-occlusive disease (n=22), or AAAs (n=50) at the Helsinki University Central Hospital. RESULTS: The median levels of cLPS in plaques were 2.28, 0.80, and 0.29 ng/ml in AAA, aorto-occlusive disease, and carotid artery stenosis patients, respectively (P<.001, Kruskal-Wallis). cLPS in serum correlated with LPS binding protein levels (Spearman's rho=0.52, P<.001), suggesting that the presence of chlamydiae is sufficient to produce an innate immune response reaction in these patients. Serum inflammatory markers interleukin 6 and highly sensitive C-reactive protein also correlate with cLPS (Spearman's rho=0.42 and 0.51, respectively, P<.001). CONCLUSIONS: cLPS is present in arterial disease, and the potential role of C. pneumoniae in the pathogenesis of both peripheral atherosclerosis disease and AAA should not be forgotten. cLPS has a positive correlation with serum inflammatory markers, but this is no proof of a causal association.

Extracranial infected carotid artery aneurysm.

Infected aneurysms of the extracranial carotid arteries are rare. This is a case report of a 63-year-old female who developed an infected internal carotid artery aneurysm due to group B Streptococcus ten days after a dental procedure. She was successfully treated with excision of the aneurysm and common to internal carotid artery bypass with greater saphenous vein.

Amalgamation of Chlamydia pneumoniae inclusions with lipid droplets in foam cells in human atherosclerotic plaque.

Chlamydia pneumoniae (Chlamydophila pneumoniae) infect macrophages and accelerates foam cell formation in in vitro experiments, but whether this might occur in human atherosclerosis is unknown. In the present study, we examined 17 carotid artery segments, obtained by endarterectomy, in which the presence of C. pneumoniae was confirmed by both polymerase chain reaction and immunohistochemistry. Electron microscopy demonstrated the presence of structures with the appearance of elementary, reticulate and aberrant bodies of C. pneumoniae in the cytoplasm of macrophage foam cells. The volume of the cytoplasm that was free from vacuoles and lipid droplets in C. pneumoniae-infected foam cells was dramatically reduced, and a phenomenon of the amalgamation of C. pneumoniae inclusions with lipid droplets was detected. Double immunohistochemistry showed that C. pneumoniae-infected foam cells contained a large number of oxidized low-density lipoproteins. The observations provide support to the hypothesis that C. pneumoniae could affect foam cell formation in human atherosclerosis.

Comparison of polymerase chain reaction methods, in situ hybridization, and enzyme immunoassay for detection of Chlamydia pneumoniae in atherosclerotic carotid plaques.

Chlamydia pneumoniae has been associated with cardiovascular diseases and has been shown by different methods to be present in atherosclerotic lesions. However, not all studies have found C. pneumoniae in atherosclerotic tissues. We compared polymerase chain reaction (PCR) methods, in situ hybridization (ISH), and measurement of chlamydial lipopolysaccharide (cLPS) by enzyme immunoassay (EIA) from homogenized atherosclerotic tissue in the detection of C. pneumoniae. In a study population of 110 patients with carotid artery disease, cLPS was found in 22.2%, and DNA by PCR was found in 34.3% and by ISH in 39.4% of the samples. Poor repeatability was shown to complicate PCR, and the technical problems inherent in ISH were not insignificant. In contrast, the cLPS EIA test was fast and easy to perform. If the sensitivity could be increased, for example, by testing multiple tissue pieces, cLPS EIA might provide a standardized commercial method for the detection of chlamydia in tissue samples, and it, thus, merits further characterization and validation in different patient populations.

Microbiologic analysis of periodontal pockets and carotid atheromatous plaques in advanced chronic periodontitis patients.

BACKGROUND: In recent years, many researchers have focused their attention on the ability of periodontal pathogens to colonize atheromatous plaques. Nevertheless, a clear correlation between the detection rates of periodontopathic bacterial DNA in atheromas and in subgingival plaque samples has not been established. The aim of our study was to assess the presence of five periodontal pathogens (Actinobacillus actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia [formerly Tannerella forsythensis]) in periodontal pockets and in carotid atheromas recovered from the same patients. METHODS: Thirty-three patients with advanced chronic periodontitis scheduled for endarterectomy were enrolled in the study. DNA was extracted from subgingival plaque samples and carotid atheromas. Universal bacteria primers for general detection of bacteria and species-specific primers for detection of periodontal pathogens were used to amplify part of the 16S rRNA gene by polymerase chain reaction. RESULTS: All subgingival plaque samples were positive for at least one target microorganism. The prevalence of T. forsythia, P. gingivalis, T. denticola, P. intermedia, and A. actinomycetemcomitans were 69.7%, 63.6%, 54.5%, 45.4%, and 33.3%, respectively. Bacterial DNA was detected in 31 out of 33 endarterectomy specimens. However, none of the samples tested positive for DNA from periodontal pathogens. CONCLUSION: The presence of periodontal bacteria in atheromatous plaques was not confirmed by this investigation; thus, no correlation could be drawn between periodontitis bacteria and microorganisms involved in the atherosclerotic lesions.

Lack of association between infectious burden and carotid atherosclerosis in Japanese patients.

Several infectious agents, such as Chlamydia pneumoniae, cytomegalovirus (CMV), herpes simplex virus (HSV), and Helicobacter pylori, have been implicated in the pathogenesis of atherosclerosis; however, but the contribution of infection may vary among races and geographic conditions. The present study investigates the association between the presence of these pathogens and carotid atherosclerosis and examines the relevance of an infectious burden during atherogenesis in Japanese patients undergoing carotid endarterectomy. We investigated a total of 50 carotid atherosclerotic plaques resected during carotid endarterectomy by polymerase chain reaction (PCR) for C. pneumoniae, CMV, HSV, and H. pylori and by immunocytochemistry (ICC) for C. pneumoniae. We also examined the presence of antibodies to IgG and/or IgA for each pathogen in blood samples. We detected HSV DNA in 2 specimens (4%) and positive ICC for C. pneumoniae in 8 (16%). The results of PCR, ICC, or serum antibodies, as well as the number of seropositive antibodies, did not correlate with severely stenotic, ulcerative, or symptomatic plaques. Our findings indicate that the detection rate of infectious agents within atherosclerotic plaques was significantly lower in our patients than that in other studies. Thus, an inflammatory mechanism might not correlate with the pathogenesis of carotid atherosclerosis among Japanese patients with severe carotid artery stenosis.

Periodontal pathogens in periodontal pockets and in carotid atheromatous plaques.

AIM: In the last years the relationship between periodontitis and cardiovascular diseases has been a subject of increasing research. The identification of periodontopathic bacteria in atheromas can contribute to our knowledge of such an association. The aim of our study was to assess the concomitant presence of 5 periodontal pathogens (Actinobacillus actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia) in periodontal pockets and in carotid atheromas recovered from the same individuals. METHODS: Twenty-one patients with chronic periodontitis scheduled for endarterectomy for carotid stenosis were enrolled in the study. Subgingival plaque samples and carotid atheromas were examined using the polymerase chain reaction and reverse hybridization techniques by means of specific probes for periodontal bacteria. Human beta-globin amplification was used as internal polymerase chain reaction efficiency control. RESULTS: Three patients were excluded because the endarterectomy specimens were negative to DNA amplification. All subgingival plaque samples were positive for at least one target micro-organism. The prevalence of Tannerella forsythia, Porphyromonas gingivalis, Prevotella intermedia, Treponema denticola, and Actinobacillus actinomycetemcomitans was 72.22%, 61.11%, 55.56%, 50%, and 33.33%, respectively. No periodontal bacteria DNA was detected in any endarterectomy specimen. CONCLUSIONS: The presence of periodontal bacteria in atheromatous plaques was not confirmed by this investigation and, thus, no correlation between periodontitis bacteria and micro-organisms involved in the atherosclerotic lesions could be drawn.

No detectable Chlamydia pneumoniae and cytomegalovirus DNA in leukocytes in subjects with echolucent and echogenic carotid artery plaques.

BACKGROUND: Controversy exists whether persistent Chlamydia pneumoniae or cytomegalovirus infections cause initiation or progression of atherosclerosis. C. pneumoniae DNA in peripheral blood mononuclear cells (PBMC) has been proposed to be a more reliable marker of cardiovascular risk than are C. pneumoniae antibodies. Reported prevalences of C. pneumoniae DNA among cardiovascular patients vary greatly, indicating methodological limitations. There is an increasing concern that published results may have been biased by extensive use of less specific polymerase chain reaction (PCR) technology. METHODS: C. pneumoniae DNA and cytomegalovirus DNA were determined by probe-based real-time PCR technology in PBMCs among subjects with echolucent (n=29) or echogenic (n=28) carotid artery plaques, and in controls without carotid plaques (n=38), all recruited from a population-based study. Samples were examined in multiple repeats with PCR assays targeting two different sequences of the genome for both microorganisms. RESULTS AND CONCLUSION: IgG seropositivity was frequent in all three groups, confirming previous exposure, but C. pneumoniae DNA or cytomegalovirus DNA was not detected in a single PBMC sample by means of probe-based, highly sensitive, and specific real-time PCR assays. Our results indicate that persistent C. pneumoniae or CMV infection is not a common phenomenon in subjects with carotid atherosclerosis.

The association of carotid plaque inflammation and Chlamydia pneumoniae infection with cerebrovascular symptomatology.

OBJECTIVE: Inflammation and infection have been implicated in the pathogenesis of carotid artery atherosclerosis, but their role in cerebrovascular disease symptomatology is not so well defined. We hypothesized that carotid disease symptomatology was associated with specific serologic markers of inflammation and Chlamydia pneumoniae infection and the presence of the pathogen and concentration of tumor necrosis factor-alpha (TNF-alpha) on the atheroma. METHODS: In 2004, 78 patients underwent carotid endarterectomy in our department, 46 of whom were symptomatic (group A) and 32 were asymptomatic (group B). A detailed medical history, the presence of atherosclerosis risk factors, ankle-brachial index and boxy mass index were recorded. We measured preoperatively the levels of C-reactive protein, fibrinogen, TNF-alpha, and the titers of immunoglobulin (Ig) A and IgG antibodies against C pneumoniae in the serum. Finally, the atherosclerotic plaques of all patients were immunohistochemically examined for the presence of C pneumoniae and their TNF-alpha concentration was determined. RESULTS: Isolation of the pathogen on the atheromatous lesion was statistically correlated with several risk factors and some of the variables that were tested. After testing independence of association, using the multiple regression analysis, only male gender (P = .024), hypertension (P = .008), hypercholesterolemia (P = .001), and TNF-alpha plaque values (P = .008) remained significantly associated. Hypertension, serum levels of fibrinogen and anti-C pneumoniae immunoglobulin A, detection of C. pneumoniae on the plaque, and plaque TNF-alpha values were significantly correlated with carotid disease symptomatology. After multiple analyses, only the presence of the pathogen on the lesion (P = .008) and atheroma TNF-alpha levels (P = .025) remained significantly associated with cerebrovascular events. CONCLUSIONS: It seems that hypertensive hypercholesterolemic men are more likely to have C. pneumoniae infected carotid plaques and that these plaques have higher TNF-alpha concentrations. Cerebrovascular disease symptomatology is strongly correlated with both C. pneumoniae infection and TNF-alpha concentration of the atheroma.