47 research outputs found
A Krónikus Chlamydia Fertőzés Transzkriptom Alapú Diagnózisa és Kezelése - ChlamyTrans - = Transcriptome-based Monitoring and Eradication of Chronic Chlamydial Infection - ChlamyTrans -
A ChlamyTrans project keretében sikerült kidolgoznunk egy új típusú, PCR amplifikáción alapuló Chlamydiális mRNS amplifikációs módszert. A módszer szenzitivitását és specificitását részletesen teszteltük QPCR módszerrel. Az amplifikációs módszert Chlamydia pneumoniae-val fertőzött egértüdőben lezajló bakteriális génexpresszió monitorozására használtuk fel. Több olyan gént-géncsoportot azonosítottunk, amelyek in vivo a baktérium aktív vagy perzisztens növekedésében szerepet játszhatnak. A bakteriális génexpresszió kutatása mellett a Chlamydia fertőzés hatására bekövetkezett gazdasejti génexpressziót is monitoroztuk. Olyan géneket azonosítottunk, amelyek szerepet játszhatnak a Chlamydia fertőzések által indukált szöveti pathológia kialakításában. Végezetül olyan módszerfejlesztésekben is részt vettünk, amelyekkel a Chlamydia fajok funkcionális genetikáját lehet részletesen tanulmányozni. | In the framework of ChlamyTrans project, we developed a novel PCR-based Chlamydial mRNA amplification method. The sensitivity and specificity of the method was extensively tested using the QPCR method. The amplification was used to monitor Chlamydial gene expression in Chlamydia pneumoniae infected mouse lung tissue. The gene expression screen identified Chlamydial genes that were involved in the active and persistent growth of C. pneumoniae in vivo. Besides Chlamydial gene expression profiling we monitored the host gene expression during Chlamydia infection. We could identify host genes that could potentially participate in Chlamydia-indued tissue pathology. Finally, we developed two methods with which we can characterize the functional genetics of Chlamydiae
Chlamydia pneumoniae in atherosclerotic middle cerebral artery
Background and Purpose—Atherosclerotic middle cerebral arteries are frequent sites of thrombosis, leading to stroke. Previous studies have suggested a role for Chlamydia pneumoniae in the pathogenesis of atherosclerosis. However, the presence of this pathogen in atherosclerotic middle cerebral arteries has heretofore not been documented. In the present study, we analyzed atheromatous plaques from middle cerebral arteries for the presence of C pneumoniae.
Methods—Atherosclerotic middle cerebral arteries from 15 cadavers who died of natural causes and corresponding
nonatherosclerotic arteries from 4 otherwise healthy trauma victims were examined. Assays for C pneumoniae DNA
were carried out by nested polymerase chain reaction (nPCR) specific for the C pneumoniae ompA gene. The presence
of the bacterium was assessed by transmission electron microscopy.
Results—Five of the 15 atherosclerotic arterial samples and none of the control tissues were positive for C pneumoniae
by nPCR. Particles similar in morphology and size to C pneumoniae elementary bodies were detected by transmission
electron microscopy in 4 of the 5 nPCR-positive atherosclerotic samples.
Conclusions—The demonstration of C pneumoniae in atherosclerotic middle cerebral arteries is consistent with the hypothesis that this bacterium is involved in acute and chronic cerebrovascular diseases
High dynamic range detection of Chlamydia trachomatis growth by direct quantitative PCR of the infected cells
N-acetyl-cysteine increases the replication of Chlamydia pneumoniae and prolongs the clearance of the pathogen from mice
PURPOSE: Within the community, 10 % of acquired pneumonia is caused by Chlamydia pneumoniae. N-acetyl-cysteine (NAC) is one of the most commonly used mucolytics in respiratory diseases, but its effect on C. pneumoniae infection has not yet been investigated. In this study, our aim was to investigate whether NAC influences the replication of C. pneumoniae. After determining that NAC does have an effect on C. pneumoniae replication, the effect of an alternative drug called Ambroxol (Ax) was investigated. METHODOLOGY: The in vitro effect of NAC and Ax was studied on C. pneumoniae-infected A549 and McCoy cells. Furthermore, the influence of NAC and Ax was examined in mice infected intranasally with C. pneumoniae. RESULTS: NAC treatment resulted in approximately sixfold more efficient C. pneumoniae growth in tissue culture compared to the untreated control cells, and this effect was shown to be based on the increased binding of the bacterium to the host cells. The C. pneumoniae-infected mice to which NAC was given had prolonged and more severe infections than the control mice. Ax decreased C. pneumoniae replication in vitro, which was partially associated with the increased expression of indolamine 2,3-dioxygenase. In animals, using the adapted usual human dose, Ax did not alter the number of recoverable C. pneumoniae. CONCLUSION: Based on our results, it might be recommended that a mucolytic agent other than NAC, such as Ax, be used in respiratory diseases suspected to be caused by C. pneumoniae
Independent and joint effects of antibodies to human heat-shock protein 60 and Chlamydia pneumoniae infection in the development of coronary atherosclerosis
Background—Studies have suggested that the prevalence of antibodies against heat-shock proteins (HSPs), Chlamydia
pneumoniae (Cpn), and cytomegalovirus (CMV) is associated with coronary artery disease (CAD), but the independent
or joint effects of human (h) HSP60 antibodies and these pathogens in patients have not been fully elucidated.
Methods and Results—A total of 405 subjects (276 patients with CAD and 129 control individuals) were tested for serum
antibodies to hHSP60, Cpn, and CMV immediate-early-1 (IE1) antigens. Patients were also assessed for serum cholesterol, triglyceride levels, and smoking habit. Significantly elevated levels of antibodies to hHSP60 and Cpn but
not to CMV-IE1 antigens were documented in CAD patients. Multiple logistic regression analysis and subanalyses of
selected subjects showed that these associations were independent of age, sex, smoking, and serum lipid levels.
Antibodies to hHSP60 and Cpn did not correlate quantitatively; however, the relative risk of disease development was substantially increased in subjects with high antibody levels to both hHSP60 and Cpn, reaching an odds ratio of 82.0 (95% CI 10.6 to 625.0).
Conclusions—High levels of antibodies to hHSP60 and Cpn are independent risk factors for coronary atherosclerosis, but
their simultaneous presence substantially increases the risk for disease development
Correlation between detergent activity and anti-herpes simplex virus-2 activity of commercially available vaginal gels
Liposomal Encapsulation Increases the Efficacy of Azithromycin against Chlamydia trachomatis
Chlamydia trachomatis (C. trachomatis) is an obligate intracellular bacterium linked to ocular and urogenital infections with potentially serious sequelae, including blindness and infertility. First-line antibiotics, such as azithromycin (AZT) and doxycycline, are effective, but treatment failures have also been reported. Encapsulation of antibiotics in liposomes is considered an effective approach for improving their local effects, bioavailability, biocompatibility and antimicrobial activity. To test whether liposomes could enhance the antichlamydial action of AZT, we encapsulated AZT in different surface-charged elastic liposomes (neutral, cationic and anionic elastic liposomes) and assessed their antibacterial potential against the C. trachomatis serovar D laboratory strain as well as the clinical isolate C. trachomatis serovar F. A direct quantitative polymerase chain reaction (qPCR) method was used to measure chlamydial genome content 48 h post infection and to determine the recoverable chlamydial growth. All the liposomes efficiently delivered AZT to HeLa 229 cells infected with the laboratory Chlamydia strain, exhibiting the minimal inhibitory concentrations (MIC) and the minimal bactericidal concentrations (MBC) of AZT even 4-8-fold lower than those achieved with the free AZT. The tested AZT-liposomes were also effective against the clinical Chlamydia strain by decreasing MIC values by 2-fold relative to the free AZT. Interestingly, the neutral AZT-liposomes had no effect on the MBC against the clinical strain, while cationic and anionic AZT-liposomes decreased the MBC 2-fold, hence proving the potential of the surface-charged elastic liposomes to improve the effectiveness of AZT against C. trachomatis