Annexin A1 expression in atherosclerotic carotid plaques and its relationship with plaque characteristics.

OBJECTIVE: Annexin A1, a calcium and phospholipid-binding protein, is an important endogenous modulator of inflammation. Whether this regulatory role extends to atherosclerosis is unknown. The aim of this study is to investigate the genetic and protein expression of Annexin A1 in carotid endarterectomy specimens from patients with significant carotid stenosis. MATERIALS AND METHODS: The echogenicity of atherosclerotic plaques was determined by ultrasound prior to carotid endarterectomy (CEA) in 34 consecutively recruited patients with carotid stenosis exceeding 70%. The Annexin A1 messenger RNA and protein expression of the corresponding plaques obtained from those patients were analyzed by reverse transcriptase polymerase chain reaction (RT-PCR) and the immunohistochemical method respectively. Results were analyzed with respect to plaque characteristics and symptomatic disease. RESULTS: There were 25 males and 9 females, with a mean age of 68.8. Ten patients were asymptomatic. The symptomatic patients' plaques were more echolucent (mean grey scale median (GSM) of 103) than those of asymptomatic patients (mean GSM = 126, p = 0.022). The Annexin A1 protein was constitutively expressed in all plaques, and Annexin A1 gene expression was statistically higher in patients with asymptomatic disease compared with those with neurological symptoms (87 ± 4% vs. 42 ± 6.2%; p < 0.001, unpaired t-test). The GSM score was positively correlated with Annexin A1 levels in patients with high-grade carotid artery stenosis (r = 0.501, p = 0.009). CONCLUSIONS: This is the first study to suggest that high Annexin A1 expression may have a stabilising effect in asymptomatic patients with less echolucent atherosclerotic plaques. Since atherosclerosis is an inflammatory process, we further postulate that Annexin A1 may play an essential role in preventing plaque complications or disease progression.

Detection of C. pneumoniae by polymerase chain reaction-enzyme immunoassay in abdominal aortic aneurysm walls and its association with rupture.

OBJECTIVE: Serological studies have suggested that one of the risk factors for aneurysm development is C. pneumoniae infection. The purpose of this study was to evaluate whether there is an association between the presence of C. pneumoniae DNA in aneurysms and ruptured abdominal aortic aneurysms. METHODS: Aortic walls were collected consecutively from 30 patients with intact AAA, 16 patients with ruptured AAA and 19 healthy organ donors (control). Purified DNAs from all aortas were analyzed for the presence of C. pneumoniae DNA in parallel by polymerase chain reaction-enzyme immunoassay (PCR-EIA) and agarose gel electrophoresis. PCR-EIA has a high sensitivity in detecting low DNA copy number in clinical atherosclerotic samples. RESULTS: C. pneumoniae DNA was detected more frequently in patients with aneurysms, particular with ruptured aneurysms. The incidence of positive C. pneumoniae DNA was 73.3% in intact AAA and 10.5% in control aortas, with the highest frequency in ruptured AAA (100%) (p < 0.05). CONCLUSION: Giving the high specificity and sensitivity of PCR-EIA, these findings support the association of C. pneumoniae in the pathogenesis of aneurysm development, growth and rupture.

Epidemiology of venous thromboembolism in a Chinese population.

BACKGROUND: Deep vein thrombosis (DVT) is uncommon in Asians and routine thromboprophylaxis for surgery is controversial. Despite recent reports of higher incidences in some Asian countries, population-based data are lacking. METHODS: Information from 2000 to 2001 was retrieved from a centralized computer public healthcare database serving an ethnic Chinese population of 6.7 million. The incidence, demographics and hospital mortality rates of DVT and pulmonary embolism (PE) were obtained, and analysed for different surgical categories. RESULTS: The overall annual incidences of DVT and PE were 17.1 and 3.9 per 100000 population respectively. Venous thromboembolic disease was more common with increasing age in both sexes. The annual age-specific incidences of DVT and PE were 81.1 and 18.6 per 100000 for those aged 65 years and over. Hospital mortality rates associated with DVT and PE were 7.3 and 23.8 per cent respectively. Among 120940 surgical operations a year, the mean incidence of postoperative DVT and PE was only 0.13 and 0.04 per cent respectively. No high-risk surgical group was identified. CONCLUSION: Venous thromboembolism is not as common in Chinese as in Caucasians, but it is certainly not rare. The majority of DVTs and PEs, however, were not associated with surgery, so routine thromboprophylaxis may not be required.

COX-dependent and -independent pathways in bradykinin-induced anion secretion in rat epididymis.

Lysylbradykinin (LBK) added to the apical or basolateral side of cultured rat epididymal monolayers stimulated a rise in short-circuit current (Isc) due to anion secretion. The concentration-response relationships for the apical and basolateral applications have EC50 value of 0.001 microM. The responses to apical or basolateral application of LBK were blocked by WIN64338, a specific B2 receptor antagonist, but not by Des-Arg9,[Leu8]-BK, a specific B1 receptor antagonist, indicating that the LBK effects were mediated through B2 bradykinin receptors. Experiments to desensitize the B2 receptors by repeated stimulation have demonstrated that the responses to apical or basolateral LBK were due to discrete receptors on the apical or basolateral surface. In epithelia clamped in the Ussing chambers, addition of LBK to the apical or basolateral surface evoked release of PGE2 into the apical and basolateral bathing solutions over the first 10 min following hormone addition. LBK added to the basolateral side elicited a greater release than it was added to the apical side. Pretreatment of the epithelia with piroxicam (5 microM) abolished PGE2 release elicited by apical or basolateral LBK and abrogated the Isc induced by basolateral LBK. However, the rise in Isc induced by apical LBK was reduced by 31.3% only. The anion secretion response to apical LBK was not affected by MDL-12330A, an adenylate cyclase inhibitor, but greatly attenuated by thapsigargin, an inhibitor of intracellular Ca2+ release. However, the reverse effects were seen for basolateral LBK. It is concluded that distinct pathways are involved in the stimulation of anion secretion by apical or basolateral LBK. The response to basolateral LBK was COX-dependent, mediated by PGE2 and involves cAMP as second messenger. In contrast, the response to apical LBK is largely COX-independent, not mediated by PCE2 and involves Ca2+ as intracellular messenger.

Cyclooxygenase-2 regulates apoptosis in rat epididymis through prostaglandin D2.

In previous studies, cyclooxygenase (COX)-1 and COX-2 isozymes have been detected in the rat epididymis. COX-1 mediates electrolyte and fluid secretion induced by a number of peptide hormones, including bradykinin, angiotensin, and endothelin, via local formation of prostaglandin (PG) E2; however, the physiological role of COX-2 remains largely unknown. Marked apoptotic cell death in the rat epididymis following androgen depletion has been reported. Because expression of both COX isozymes is dependent on androgen, we investigated whether these isozymes control apoptosis in the epididymis. Apoptosis was detected in rat epididymal epithelial cells by in situ staining using the TUNEL method and by the presence of internucleosomal DNA fragmentation using capillary electrophoresis with laser-induced fluorescence detection. Specific COX inhibitors were used to delineate the roles of the 2 isozymes. There was no significant apoptotic cell death in normal and specific COX-1 inhibitor (SC-560)-treated epididymal cells. However, application of a specific COX-2 inhibitor (NS-398) induced apoptosis in a dose- and time-dependent manner. A similar apoptotic effect of COX-2 inhibitor was seen in the in vivo study. The drastic DNA fragmentation induced by COX-2 inhibitor could be reversed completely by PGD2 and partially by PGE2. In addition, the protective effect of PGD2 against COX-2 inhibition was significantly blocked by a PGDP-receptor antagonist, BWA868C. These results indicate that the COX-2 products PGD2 and, to a lesser extent, PGE2 control apoptosis in cultured rat epididymal cells in vitro.