eduction compared to single-dose irradiation. However, a more profound decrease in IFU/mL was observed after threefold irradiation, compared to that observed after a single dose of irradiation ). Because these data indicate that multiple irradiations have an additive effect on chlamydial infectivity, we examined the ultrastructure of threefold-irradiated chlamydial inclusions. HeLa cells were infected, irradiated three times, fixed after a 3-hour incubation period and further processed for electron microscopy at 43 hpi. We counted the total number of bacteria in irradiated and non-irradiated, Chlamydia-infected cells as previously described. Containing 4627 chlamydial bodies in total, nonirradiated inclusions consisted of more than twice as many bacteria as irradiated inclusions. Similar to single-dose irradiation, chlamydial developmental forms were categorized into different developmental stages according to their morphology. Irradiated inclusions contained fewer EBs 12695532 but a higher amount of RBs 7 wIRA/VIS Inhibits Chlamydia compared to the non-irradiated chlamydial inclusions with 31.3%613.48% EBs and 45.3%612.45% RBs, respectively. The distribution of IBs was similar in both conditions with 23.2%66.85% in non-irradiated and 22.8%66.91% in irradiated inclusions, respectively. The number of ABs was negligible in both testing conditions. Irradiation and chlamydial infection trigger an identical subset of released cytokines and chemokines from cells Finally, to investigate the response of HeLa cells to irradiation, to chlamydial infection, and to the combination of both, cytokine and chemokine assays were performed. This experiment included four different treatment groups: non-infected HeLa cells without irradiation, non-infected HeLa monolayers with irradiation, 8 wIRA/VIS Inhibits Chlamydia C. trachomatis-infected HeLa cultures without irradiation, and C. trachomatis-infected HeLa cultures with irradiation. Triple-dose irradiation was performed as 22408714 previously GW788388 price described, the supernatants of each experimental set were collected at 43 hpi, and cytokines and chemokines were analyzed using cytokine and chemokine array panel kits according to manufacturer’s instructions. Beforehand, the arrays were validated by assessment of the linearity of internal assay controls determined over time. We observed a similar release pattern in five out of 20 cytokines and in five out of 24 chemokines in all three treatment groups -. However, expression of most cytokines and chemokines tested did not change under the treatment conditions examined. Discussion Antimicrobial therapy is the treatment of choice for bacterial infections. However, all pharmacological approaches bear the risk wIRA/VIS Inhibits Chlamydia of unwanted side effects such as antibiotic-associated diarrhea in patients treated with amoxicillin-clavulanate, or more severe risks such as cephalosporin-induced immune hemolytic anemia, drug-induced liver toxicity due to various antimicrobial agents and kidney injury caused by aminoglycosides, vancomycin and others. Chlamydia trachomatis is a known agent of ocular and genital infections leading to blindness and sexually transmitted diseases. Recommended antibiotic therapies to treat these conditions are either by multiple doses of tetracycline or a single dose of azithromycin. However, a recent study has found that the risk of cardiovascular death in patients on azithromycin is elevated, compared to patients treated with amoxicillin or ciprofloxacin