Microbial and vaginal determinants influencing Mycoplasma hominis and Ureaplasma urealyticum genital colonization in a population of female patients.

Mycoplasma hominis and Ureaplasma urealyticum are associated with chorioamnionitis, preterm delivery and pelvic inflammatory disease. The aim of this study was to evaluate the possible risk factors of co-colonization by M. hominis in patients already colonized by U. urealyticum and compare demographic parameters, vaginal pH and microbiota of women colonized by U. urealyticum or M. hominis. A total of 452 patients positive for U. urealyticum or M. hominis were analysed, 421 (93.1%) of whom were positive for U. urealyticum and 31 (6.9%) for M. hominis. Patients positive for M. hominis compared to patients positive for U. urealyticum were more frequently colonized by Gardnerella vaginalis (71% vs 18.5%; p 0.0001), less frequently by lactobacilli (16.1% vs 61.5%; p 0.0001), and more frequently had a pH value higher than 4.5 (96.8% vs 57%; p 0.0001), all conditions associated to bacterial vaginosis (BV). Logistic regression analysis showed that only G. vaginalis colonization and pH higher than 4.5 were independently related to M. hominis colonization (respectively p 0.0001 and p 0.016). Thus, in women colonized by U. urealyticum, BV is an independent risk factor for M. hominis co-colonization.

Prevalence and antimicrobial susceptibility of Ureaplasma urealyticum and Mycoplasma hominis in a population of Italian and immigrant outpatients.

Ureaplasma urealyticum and Mycoplasma hominis are associated with non-gonococcal urethritis, increased risk of recurrent miscarriage, infertility and pelvic inflammatory disease. Migration flows from other countries change the local epidemiological profile of infectious diseases of patients treated by general practitioners and hospital doctors. Few studies have evaluated this ever-changing issue in the Italian population. The aim of this study was to assess possible differences in prevalence and antimicrobial susceptibility of U. urealyticum and M. hominis in a population of 433 Italian and immigrant outpatients by means of the commercially available MYCOFAST(®) Screening EvolutioN 3 Kit. Prevalence of positive samples was 44.5% in Italian patients and 53.4% in immigrants. Samples positive for U. urealyticum and total isolates were more frequent in African patients: U. urealyticum, 51.5% vs 33.3%; Yates-corrected chi-square=3.98; p=0.046; total isolates, 54.5% vs 34.3%; Yates-corrected chi-square=4.45; p=0.035. Among samples positive for U. urealyticum, 66.4% were resistant to ciprofloxacin, whereas 27.6% to ofloxacin. In M. hominis isolates, 66.7% were resistant to both azythromycin and roxythromycin. Our study showed how prevalence of genital mycoplasmas and antibiotic resistance profiles change in relation to the country of origin. Therefore, surveillance is critical for the early cure and prevention of the occurrence of resistant strains.

Procalcitonin predicts real-time PCR results in blood samples from patients with suspected sepsis.

BACKGROUND: Early diagnosis and rapid bacterial identification are of primary importance for outcome of septic patients. SeptiFast® (SF) real-time PCR assay is of potential utility in the etiological diagnosis of sepsis, but it cannot replace blood culture (BC) for routine use in clinical laboratory. Procalcitonin (PCT) is a marker of sepsis and can predict bacteremia in septic patients. The aim of the present study was to investigate whether PCT serum levels could predict SF results, and could help screening febrile patients in which a SF assay can improve the etiological diagnosis of sepsis. METHODS: From 1009 febrile patients with suspected sepsis, 1009 samples for BC, SF real-time PCR, and PCT determination were obtained simultaneously, and results were compared and statistically analysed. Receiver operating characteristic (ROC) curves were generated to determine the area under the curve and to identify which cut-off of PCT value produced the best sensitivity to detect SF results. RESULTS: Mean PCT values of sera drawn simultaneously with samples SF positive (35.42 ± 61.03 ng/ml) or BC positive (23.14 ± 51.56 ng/ml) for a pathogen were statistically higher than those drawn simultaneously with SF negative (0.84 ± 1.67 ng/ml) or BC negative (2.79 ± 16.64 ng/ml) samples (p<0.0001). For SF, ROC analysis showed an area under the curve of 0.927 (95% confidence interval: 0.899-0.955, p<0.0001). The PCT cut-off value of 0.37 ng/ml showed a negative predictive value of 99%, reducing the number of SF assays of 53.9%, still identifying the 96.4% of the pathogens. CONCLUSION: PCT can be used in febrile patients with suspected sepsis to predict SF positive or negative results. A cut-off value of 0.37 ng/ml can be considered for optimal sensitivity, so that, in the routine laboratory activity, SF assay should not be used for diagnosis of sepsis in an unselected patient population with a PCT value <0.37 ng/ml.

Comparison of conventional culture with SeptiFast real-time PCR for microbial pathogen detection in clinical specimens other than blood.

Early detection of aetiological agents is pivotal for adequate therapy for bacterial infections. Although culture is still considered the mainstay for laboratory diagnosis, it often lacks sensitivity, especially in patients already treated with antibiotics. The present study investigated the potential clinical utility of the commercial real-time-PCR-based system SeptiFast (SF), originally intended for diagnosis of sepsis from blood specimens, in the aetiological diagnosis of other bacterial infections, in patients undergoing antibiotic therapy. A total of 53 non-blood specimens were analysed for microbial pathogen detection by conventional culture and with SF real-time PCR: 19 (35.8%) synovial fluids, 9 (17.0%) cardiac valve tissues and 25 (47.2%) purulent exudates from various body sites. Overall, the number of specimens positive for a pathogen by SF (26/53; 49.1%) was significantly greater (P=0.001) than that of specimens positive by culture (10/53; 18.9%). In particular, SF was superior to culture for pathogen detection in cardiac valve tissues and synovial fluids. The analysis of concordance showed a fair agreement between the two methods (kappa value=0.314; 95% confidence interval=0.531-0.097). Even with the limitation of the low number of specimens, this study confirmed the great potential of diagnosing bacterial infections by a molecular approach, and indicates that the real-time PCR SF system can be used for specimens other than blood, from patients undergoing antibiotic treatment.

Antifibrogenic effects of canrenone, an antialdosteronic drug, on human hepatic stellate cells.

BACKGROUND & AIMS: Several lines of evidence indicate that aldosterone antagonists may exert direct antifibrogenic effects. The aim of this study was to evaluate the possible direct antifibrogenic effects of canrenone, the active metabolite of spironolactone, in activated human hepatic stellate cells. METHODS: The effects of canrenone were assessed on platelet-derived growth factor-induced mitogenic and chemotactic effects and the increased de novo synthesis of different extracellular matrix components induced by transforming growth factor-beta1. RESULTS: Canrenone dose-dependently reduced platelet-derived growth factor-induced cell proliferation and motility. This effect was not associated with either changes in the phosphorylation of platelet-derived growth factor receptor and phospholipase C gamma or in the activation of the Ras/extracellular signal-regulated kinase pathway, whereas it was accompanied by a dose-dependent inhibition of platelet-derived growth factor-induced phosphatidylinositol 3-kinase activity. In addition, canrenone inhibited the activity of the Na(+)/H(+) exchanger 1 induced by platelet-derived growth factor. The effect of canrenone on Na(+)/H(+) exchanger 1 activity was reproduced by phosphatidylinositol 3-kinase inhibitors, thus supporting an inhibitory action of canrenone on phosphatidylinositol 3-kinase activity. To further address this possibility, the action of canrenone was compared with that of 2 established Na(+)/H(+) exchanger 1 inhibitors: ethylisopropylamiloride and cariporide. Whereas ethylisopropylamiloride was able to inhibit platelet-derived growth factor-induced phosphatidylinositol 3-kinase activity, cariporide was without any effect. Both compounds reproduced the effects of canrenone on platelet-derived growth factor-induced mitogenesis and chemotaxis. Finally, canrenone was able to reduce transforming growth factor-beta1-induced de novo synthesis of procollagen type I/IV and fibronectin and thrombin-induced hepatic stellate cell contraction. CONCLUSIONS: These results indicate that canrenone may be active as an antifibrogenic drug.