Provocation of experimental aortic inflammation and dilatation by inflammatory mediators and Chlamydia pneumoniae.

BACKGROUND: The macrophage appears to have a key role in the inflammation and proteolysis associated with the growth and development of abdominal aortic aneurysms. The role of inflammatory mediators and Chlamydia pneumoniae in stimulating the influx of macrophages and dilatation of the abdominal aorta was investigated in an experimental model. METHODS: Periaortic application of calcium chloride solution (and monocyte chemoattractant protein (MCP) 1, a cocktail of cytokines or C. pneumoniae) to the abdominal aorta of New Zealand White rabbits was performed at laparotomy. Some animals were fed a cholesterol-rich diet. The diameter of the aorta was measured by ultrasonography and after perfusion fixation, 3 weeks after laparotomy. Aortic sections were stained with RAM-11 to identify macrophages for counting. The presence of C. pneumoniae DNA was confirmed using the polymerase chain reaction. RESULTS: Aortic macrophage influx in response to MCP-1, thioglycollate or C. pneumoniae was more than doubled in the cholesterol-fed animals. In response to human recombinant MCP-1 (1 microg) the mean(s.d.) macrophage count increased from 79(19) to 340(215) per unit area (P < 0.02). Even in cholesterol-fed animals, application of MCP-1 (recombinant human or rabbit form) was not associated with aortic dilatation. Application of thioglycollate 0.1 mol/l, or live or formalin-inactivated C. pneumoniae (0.5 x 108 organisms), was associated with a similar increase in macrophages to that caused by MCP-1 and a significant (approximately twofold) increase in aortic diameter after 3 weeks. CONCLUSION: Macrophage influx into rabbit abdominal aorta, without macrophage activation, is insufficient to cause experimental aortic dilatation. C. pneumoniae antigens appeared to stimulate aortic dilatation, probably by specific activation of macrophages.     

Chlamydia pneumoniae and chronic bronchitis: association with severity and bacterial clearance following treatment

BACKGROUND: A study was undertaken to evaluate Chlamydia pneumoniae chronic infection, other respiratory infections, and functional impairment in patients with chronic bronchitis (stage 1) and to examine chronic C pneumoniae infection, rate of acute exacerbations of chronic bronchitis, and rate of C pneumoniae eradication following antibiotic treatment (stage 2). METHODS: In the stage 1 study respiratory specimens from 42 patients with steady state chronic bronchitis were analysed for Gram staining, sputum culture, and C pneumoniae DNA detection by nested touchdown polymerase chain reaction (PCR). On the basis of the results of stage 1, a second population of 141 consecutive patients with steady state mild to moderate chronic bronchitis (FEV(1) >or=50% predicted) was studied. On admission, at regular intervals, and at exacerbation all patients underwent serological testing for C pneumoniae (microimmunofluorescence) and a nested touchdown PCR to detect C pneumoniae DNA was performed on peripheral blood mononuclear cells (PBMCs). Patients were assessed over a 12 month period. Information regarding the previous 12 months was taken from medical records. RESULTS: Chronic colonisation of the sputum with C pneumoniae was significantly associated with lower FEV(1) and greater airway bacterial colonisation. On admission to the stage 2 study, 80 patients were PCR negative and 61 were PCR positive. Over the 2 years a mean (SD) of 1.43 (1.32) acute exacerbations occurred in PCR negative patients and 2.03 (1.21) in PCR positive patients (p<0.01). During the 12 month follow up period 34 PCR positive patients had acute exacerbations and were treated with azithromycin for 6 weeks. Serological evidence of acute C pneumoniae reinfection/reactivation was found in two of the 34 patients. The rate of C pneumoniae DNA clearance from blood following treatment was 29% at follow up. CONCLUSION: Chronic colonisation with C pneumoniae is associated with a higher rate of exacerbations of chronic bronchitis. Long term treatment is required to obtain clearance of the organism from the blood.     

Neuraminic acid content of sputum in chronic bronchitis

The neuraminic acid content of sputum from 48 men with early chronic bronchitis has been estimated in samples collected over a period of three years. The results are compared with those from 29 advanced bronchitic patients and are related to the clinical features of both groups and to the physical and biological properties of the sputum. A seasonal variation in neuraminic acid content has been noted for the first time with higher levels during the winter months. Clinical assessment of sputum pourability correlated well with measured viscosity. The viscosity of mucoid sputum was related to its neuraminic acid content but also to the yield of dry macromolecular material. In the early bronchitic group whose sputum was assessed for purulence at monthly intervals pus was more often present in those men whose mucoid sputum contained higher levels of neuraminic acid. These findings are discussed in relation to the cause of exacerbations of chronic bronchitis.     

Use of induced sputum cell counts to investigate airway inflammation in asthma.

BACKGROUND: Airway inflammation is considered to be important in asthma but is relatively inaccessible to study. Less invasive methods of obtaining sputum from patients unable to produce it spontaneously should provide a useful investigational tool in asthma. METHODS: A method to induce sputum with inhaled hypertonic saline was modified for use in 17 asthmatic patients and 17 normal subjects who could not produce sputum spontaneously. The success rate and safety of the method, the reproducibility of cell counts, and differences in cell counts between the asthmatic and normal groups were examined. Hypertonic saline solution 3-5% was inhaled for up to 30 minutes after inhalation of salbutamol. Subjects were asked to expectorate sputum every five minutes. The quality of the sample was scored on the volume of plugs and the extent of salivary contamination. Plugs from the lower respiratory tract were selected for a total cell count and for differential cell counts of eosinophils and metachromatic cells (mast cells and basophils) in direct smears. RESULTS: Adequate samples from the lower respiratory tract were obtained in 76% of first attempts. The mean fall in the forced expiratory volume in one second (FEV1) during inhalation of saline was 5.3% and the maximum fall 20%. Eosinophil and metachromatic cell counts were reproducible (reliability coefficient 0.8 and 0.7 respectively). When compared with sputum from normal subjects sputum from asthmatic patients contained a significantly higher proportion of eosinophils (mean 18.5% (SE 3.8%) v 1.9% (0.6%)) and metachromatic cells (0.50% (0.18%) v 0.039% (0.014%)). In the asthmatic group the differential eosinophil count correlated with the baseline FEV1. CONCLUSION: Induced sputum is capable of detecting differences in cell counts between normal and asthmatic subjects and merits further development as a potential means of assessing airway inflammation in asthma.     

No evidence of acute Chlamydia pneumoniae infection in patients with Bell's palsy.

The cause of Bell's palsy remains unknown even though available evidence suggests that infection could be a factor. In recent studies, Chlamydia pneumoniae has been associated with neurologic diseases such as multiple sclerosis. In the present study, the association of C pneumoniae with Bell's palsy was studied with the use of serology and polymerase chain reaction to test tear fluid and peripheral blood mononuclear cells from 21 patients with Bell's palsy and 21 control subjects. C pneumoniae DNA was detected from tear fluid samples in 1 patient with Bell's palsy and in 2 healthy control subjects. Whether this indicates earlier disease or subclinical infection remains to be studied. However, an association between Bell's palsy and acute C pneumoniae infection could not be shown.     

Serum, saliva, and sputum levels of metronidazole in acute exacerbations of chronic bronchitis.

We have evaluated the absorption and the penetration of metronidazole into the bronchial secretions and saliva in acute infective exacerbations of chronic bronchitis. Seventeen patients were given 400 mg orally three times daily for seven days and "steady state" levels were measured in serum, saliva, and sputum on the last day of treatment. Mean levels in the three biological fluids were not significantly different. Higher metronidazole levels in sputum tended to occur in patients with higher serum levels. In all but one patient, levels in serum and saliva were well within the therapeutic range. We conclude that this oral regimen results in therapeutic tissue levels in acute exacerbations of chronic bronchitis.     

Plasma and sputum erythromycin concentrations in chronic bronchitis.

Plasma and sputum concentrations of erythromycin were measured in 10 patients with chronic bronchitis during an eight-day course of a new formulation of erythromycin stearate. The plasma erythromycin levels compared favourably with the minimal inhibitory concentrations for common respiratory pathogens and indicated adequate gastrointestinal absorption when the drug was taken immediately before food. Sputum erythromycin levels were variable and in some patients low or undetectable. Measurable sputum erythromycin levels were approximately 10% of plasma levels with no evidence of accumulation and were of similar order of magnitude to the minimal inhibitory concentrations for common respiratory pathogens except Haemophilus influenzae. There was no correlation between sputum and plasma erythromycin levels. There was a trend for higher erythromycin levels in sputum containing increasing amounts of pus and also when plasma levels increased.     

Pathogenic organisms in the sputum of patients with chronic bronchitis

Bacteriological examination of the early morning sputum from 54 patients with chronic bronchitis, half of whom received chemoprophylaxis with oxytetracycline, was made regularly for periods up to four and a half years. Nasal and perineal swabs were taken for periods up to three and a half years. There was no evidence that frequent or dangerous proliferation of drug-resistant organisms occurred in the sputum of patients on prolonged chemoprophylaxis with oxytetracycline nor that a serious increase of the carriage of drug-resistant Staphylococcus pyogenes occurred in the nares or on perineal skin. In patients receiving oxytetracycline there was a significant reduction in the frequency of identification of Streptococcus pneumoniae, but the organism was not eradicated: no significant change in the frequency of identification of Haemophilus influenzae was found. From year to year there was sometimes variation in the frequency of identification of Strep. pneumoniae and Strep. pyogenes in the sputum of these patients.     

Neutrophilic inflammation and IL-8 levels in induced sputum of alpha-1-antitrypsin PiMZ subjects

BACKGROUND: Severe alpha-1-antitrypsin deficiency (AATD), due to homozygosity for the protease inhibitor (Pi) Z allele, is a genetic risk factor for chronic obstructive pulmonary disease (COPD). In a previous study the sputum of severe AATD subjects with airflow obstruction showed a pattern of cellular inflammation similar to COPD patients. It is uncertain whether heterozygotes for the Z allele or intermediate deficiency (PiMZ) have an increased risk of developing COPD. METHODS: Sputum cell counts and the supernatant level of the neutrophil chemoattractant interleukin (IL)-8 were investigated by sputum induction in 10 non-smoker asymptomatic PiMZ subjects with normal pulmonary function, 10 patients with stable COPD, and 10 age matched normal subjects. Data are expressed as mean (SD). RESULTS: The mean (SD) number of neutrophils was significantly higher (p<0.01) in the sputum of PiMZ subjects (84.5 (22.2) x10(4)/ml) and patients with COPD (126.9 (18.8) x10(4)/ml) than in matched normal subjects (55.0 (8.7) x10(4)/ml). IL-8 levels were increased in PiMZ subjects (828.5 (490.6) ng/ml; median 1003.0 ng/ml; range 1260-100 ng/ml) and in COPD patients (882.5 (524.3) ng/ml; median 934.9 ng/ml; range 1506-258 mg/ml) compared with normal subjects (3.5 (0.5) ng/ml; median 3.5 ng/ml; range 4.5-2.5 ng/ml). There was a significant positive correlation between IL-8 supernatant concentration and neutrophil count in PiMZ subjects (p = 0.036; r = 0.66). An inverse correlation was observed between the percentage of neutrophils and forced expiratory volume in 1 second (% predicted) in patients with COPD (p = 0.04; r = -0.43). CONCLUSIONS: These findings indicate that PiMZ subjects without airflow obstruction may have an IL-8 related neutrophilic inflammation in the airways, similar to stable COPD patients, suggesting an increased risk of developing pulmonary changes.     

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.     

Comparison of spontaneous and induced sputum for investigation of airway inflammation in chronic obstructive pulmonary disease

BACKGROUND: Although sputum induction is used as a technique to investigate lower airway inflammation in asthmatic subjects, advantages over spontaneous sputum in patients with chronic obstructive pulmonary disease (COPD) have not been investigated. METHODS: Samples of spontaneous sputum and sputum induced with 3% hypertonic saline for 14 minutes were collected from 27 patients with chronic obstructive pulmonary disease (COPD) who usually produced spontaneous sputum. Spirometric indices and oxygen saturation (Sao2) were measured at seven minute intervals. The spontaneous, seven and 14 minute sputum samples were analysed for total and differential cell counts, cell viability, and interleukin 8 levels. RESULTS: Analysis of the sputum revealed that median cell viability was higher in the seven minute (62.8%; p = 0.004) and 14 minute (65%; p = 0.001) induced sputum samples than in spontaneous sputum (41.2%). There was no significant difference in total and differential cell counts or in interleukin 8 levels between spontaneous and induced sputum. During the sputum induction procedure the mean (SD) fall in forced expiratory volume in one second (FEV1) was 0.098 (0.111) 1 (p < 0.001) and in forced vital capacity (FVC) was 0.247 (0.233) 1 (p < 0.001). There was a small but significant fall in Sao2 during sputum induction (p = 0.03). CONCLUSIONS: Induced sputum contains a higher proportion of viable cells than spontaneous sputum. There are no significant differences between the sputum samples obtained at seven minutes and at 14 minutes of hypertonic saline nebulisation. Sputum induction is safe and well tolerated in patients with COPD.     

Detection of Chlamydia pneumoniae in peripheral blood mononuclear cells: correlation with inflammation and atherosclerosis in haemodialysis patients.

BACKGROUND: Chlamydia pneumoniae has been implicated as an inflammatory agent in atherosclerosis. Clinical studies in this field have yielded conflicting results, which may have resulted from a lack of standardization for C.pneumoniae detection. We attempted to accurately estimate C.pneumoniae prevalence and to examine whether C.pneumoniae is associated with atherosclerosis and inflammation in haemodialysis (HD) patients. To do this, we assessed C.pneumoniae presence by a combination of methods and correlated its levels with inflammatory and atherosclerotic indexes in these patients. METHODS: Chlamydia pneumoniae was identified by polymerase chain reaction (PCR) in DNA extracted from cell cultures inoculated with patient buffy coats and by serum IgG antibodies against C.pneumoniae (IgGCp). Inflammation was assessed by C-reactive protein and serum amyloid A and atherosclerosis was evaluated from clinical and laboratory data. RESULTS: Of the 130 patients, only nine had viable C.pneumoniae in peripheral blood mononuclear cells (PBMCs) while 64 had serum IgGCp. Although patients with viable C.pneumoniae had higher atherosclerotic scores, seropositive and negative patients showed similar scores. Patients with atherosclerosis exhibited higher inflammatory indexes. Neither patients with detectable C.pneumoniae in PBMCs nor seropositive subjects had higher inflammation than negative patients. CONCLUSIONS: We found that viable C.pneumoniae in PBMCs, assessed by cell culture and PCR, was present in a small percentage of HD patients and was correlated with atherosclerosis. Seropositivity was much higher in HD patients but was not associated with viable C.pneumoniae or with atherosclerosis. Further studies in HD patients using high sensitivity and specificity methods in larger populations will be necessary to clarify the relationship between C.pneumoniae and atherosclerosis.     

Acute septic arthritis caused by Klebsiella pneumoniae

Septic arthritis caused by Gram-negative bacteria is uncommon. However, Klebsiella pneumoniae is one of the most common pathogens in Taiwan in several clinical entities, including severe community-acquired pneumonia, community-acquired lung abscess, empyema, necrotizing fasciitis, and liver abscess. However, the research focusing on septic arthritis caused by K. pneumoniae is only limited. Herein, we described three cases of K. pneumoniae-associated septic arthritis. Two of them had underlying diabetes mellitus, and one of them was caused by extended beta-lactamase–producing K. pneumoniae. All outcomes were favorable under appropriate management, which included antibiotic treatment or drainage.     

Acute flexor tenosynovitis caused by Streptococcus pneumoniae

A case of acute flexor tenosynovitis caused by Streptococcus pneumoniae in a previously well male truck driver is described. The importance of specimen collection and processing in establishing a microbiological diagnosis is emphasized. Early surgical drainage and treatment with penicillin resulted in rapid resolution of the infection and a return to full function of the hand.     

Chlamydia pneumoniae accompanied by inflammation is associated with the progression of atherosclerosis in CAPD patients: a prospective study for 3 years.

BACKGROUND: The causes of accelerated atherosclerosis in end-stage renal disease (ESRD) patients are unknown, although recent studies have suggested that Chlamydia pneumoniae (Cp) infection and inflammation might be contributing factors. We aimed to evaluate the association of carotid atherosclerosis progression with Cp infection and inflammation in patients undergoing peritoneal dialysis (PD). METHODS: This is a prospective observational study. A total of 52 non-diabetic prevalent PD patients were included. The intima-media thickness of a common carotid artery (CCA-IMT) was measured at baseline and after 36 months by B-mode ultrasonography. Serum antibodies to Cp and inflammatory markers were obtained at the time of initial measurement of the CCA-IMT. RESULTS: CCA-IMT progressors (deltaCCA-IMT > or = 0.015 mm/year) had a higher prevalence of seropositivity for Cp IgA antibody, a higher level of Cp IgA antibodies indices, log IL-(interleukin-)6, and intercellular adhesion molecule-1 (ICAM-1) compared to the non-progressors (deltaCCA-IMT < 0.015 mm/year). On multivariate analysis, Cp IgA index and log IL-6 were independent risk a factors for CCA-IMT progression. Also, Cp IgA index had independent positive correlation with the magnitude of annual deltaCCA-IMT. Cp IgA antibody seropositive patients showed significantly higher mean annual deltaCCA-IMT than seronegative patients. Moreover, patients with both positive Cp IgA antibodies and elevated IL-6 above the median level showed higher deltaCCA-IMT than those with either factor alone. CONCLUSIONS: Our data showed that Cp and inflammation were significant risk factors of CCA-IMT change in PD patients. This study strengthens evidence that Cp is involved in the pathogenesis of atherosclerosis and also suggests that the effect of Cp infection under high inflammatory status might be a risk factor for progression of atherosclerosis.