Antimicrobial Activity of N,N-Diethyldithiocarbamate against Ureaplasma parvum and Ureaplasma urealyticum.

Ureaplasma species (Ureaplasma spp.) are commonly found as commensals in the human urogenital tracts, although their overgrowth can lead to infection in the urogenital tract and at distal sites. Furthermore, ureaplasmas lack a cell wall and do not synthesize folic acid, which causes all ß-lactam and glycopeptide antibiotics, and sulfonamides and diaminopyrimidines, to be of no value. The antibiotics used in therapy belong to the fluoroquinolone, tetracycline, chloramphenicol and macrolide classes. However, the growing incidence of antibiotic-resistant Ureaplasma spp. in the population becomes a problem. Thus, there is a need to search for new drugs effective against these bacteria. Since 1951, the FDA-approved, well-tolerated, inexpensive, orally administered drug disulfiram (DSF) has been used in the treatment of chronic alcoholism, but recently, its antimicrobial effects have been demonstrated. The main biological metabolite of DSF, i.e., N,N-diethyldithiocarbamate (DDC), is generally believed to be responsible for most of the observed pharmacological effects of DSF. In the presented studies, the effect of DDC at concentrations of 2 µg/mL, 20 µg/mL and 200 µg/mL on the growth and survival of Ureaplasma urealyticum and Ureaplasma parvum was tested for the first time. The results indicated that all the used DDC concentrations showed both bacteriostatic and bactericidal activity against both tested strains.

Differences in the expression of human papillomavirus type 16 (HPV-16) E6 oncogene mRNA in SiHa cell line inoculated with CMV, HSV or ureaplasmas.

One of the factors associated with an increased risk of HPV-related malignant transformation may be bacterial and/or viral infections. The aim of our study was to examine whether the presence of infectious agents commonly detected in the genitourinary tract such as herpesviruses (HSV, CMV), and ureaplasmas (Ureaplasma urealyticum, Ureaplasma parvum) may lead to alterations in the expression of the HPV-16 E6 oncogene. Quantitative RT-PCR analysis was used to assess the level of HPV-16 E6 mRNA expression in SiHa cells. The presence of HSV-1 or HSV-2 in SiHa cells caused a 1.5-fold increase in HPV-16 E6 mRNA expression as compared with non-inoculated SiHa cells. Ureaplasma urealyticum presence but not Ureaplasma parvum stimulated the expression of HPV-16 E6 resulting in a nearly five-fold (4.8) up-regulated E6 mRNA level in SiHa cells. Our study is the first to suggest that infection of Ureaplasma urealyticum in an urogenital tract could increase the risk of cervical cancer by overexpression of the HPV E6 oncogene.

Vaginal Secretion Epithelium Count as a Prognostic Indicator of High Abundance of Ureaplasmas in Women with a Normal Nugent Score.

Genital tract ureaplasma infections are associated with numerous complications, ranging from inflammation, through infertility, to problematic pregnancy. In the course of ureaplasma infection, the risk of human papillomavirus infection increases. Diagnostic tests for urea-plasma infections are not always carried out, especially in women with the normal Nugent test results. The study attempts to check whether it is possible to find a prognostic indicator that could suggest a high abundance of ureaplasmas (≥ 104 CFU/ml) at the stage of the initial examination of vaginal discharge. Such a prognostic factor could qualify women for further tests to detect infections with these atypical bacteria. Six hundred twenty-seven white women with a score of 0-3 on the Nugent scale were tested, including 322 patients with a high abundance of ureaplasmas (≥ 104 CFU/ml) and 305 who tested negative for these bacteria. Ureaplasma infections were detected statistically significant in women who had few or no epithelial cells in the genital swab specimens compared to the results obtained for women with numerous or very numerous epithelial cells (p < 0.001). The risk of the high density of ureaplasmas was 38.7% higher with fewer or no epithelial cells than with high numbers. In patients aged 18-40 years with few or no epithelial cells, a high density of ureaplasmas (≥ 104 CFU/ml) was observed significantly more frequently (p = 0.003). Determining the number of epithelial cells in Gram-stained slides may be the prognostic indicator of ureaplasma infection. Testing for genital ureaplasma infection should be considered, especially in women of childbearing age (18-40 years), even if the Nugent test value is normal and pH ≤ 4.6.

[Evaluation of vertical transmission of two species of ureaplasmas in term newborns without respiratory disorders--a preliminary study]. / Ocena wertykalnej transmisji dwóch gatunków ureaplazm u donoszonych noworodków bez zaburzen oddechowych--badania wstepne.

UNLABELLED: Pregnancy promotes ureaplasma vaginal colonization. This creates the possibility of vertical transmission of these organisms to the child. These microorganisms can cause complications during pregnancy and poor condition of newborn. OBJECTIVES: Objectives of this study were to analyze the vertical transmission of different species of ureaplasmas in term newborns without respiratory distress. MATERIALS AND METHODS: The study included 50 mothers and 50 of their newborn children. Swabs were obtained from swabs of the cervix in women and tracheal aspirates from neonates. The presence of ureaplasmas was confirmed by culture and PCR. Ureaplasmas species identification was performed using PCR. RESULTS: infection of ureaplasmas was found in 21 women (42%). Predominant species was U. parvum, which was found in 18 women. In 3 patients only the presence of U. urealyticum was confirmed. Ureaplasma infection in mother and her newborn baby was confirmed in 8 (17.4%) mother-child pairs, including 6 of these cases showing the presence of U. parvum and 2 U. urealyticum. The incidence of vertical transmission of ureaplasma infection was assessed at 33% for U. parvum and 67% for U. urealyticum, and the total for both species at 38%. It should be noted that in the group of 18 women infected with U.parvum, in 12 cases there was no transmission of infection to the child. However in 3 women infected with U. urealyticum 2 cases of transmission from mother to child were observed (67%). Although the group infected with U. urealyticum accounted for only 3 women, our preliminary observations may suggest that this species is probably more likely to be transferred from mother to child. CONCLUSIONS: Infection with U. urealyticum may be more frequently transferred from the genital tract of mother to child.

Lipoic Acid Does Not Affect The Growth of Mycoplasma hominis Cells In Vitro.

Mycoplasma hominis is associated with various infections, for which the treatment can be complex. Lipoic acid (LA) plays a role as a cofactor in eukaryotes, most Bacteria, and some Archea. Research of recent years has increasingly pointed to the therapeutic properties of exogenously supplemented LA. The present study was conducted on 40 strains of M. hominis cultured with the following LA concentrations: 1,200 µg/ml, 120 µg/ml, and 12 µg/ml. The bacterial colonies of each strain were counted and expressed as the number of colony-forming units/ml (CFU). The number of CFU in M. hominis strains obtained in the presence of LA was compared with the number of CFU in the strains grown in the media without LA. The obtained results indicated that the presence of LA in the medium did not affect the growth of M. hominis. The investigation of the influence of LA on the growth and survival of microbial cells not only allows for obtaining an answer to the question of whether LA has antimicrobial activity and, therefore, can be used as a drug supporting the treatment of patients infected with a given pathogenic microorganism. Such studies are also crucial for a better understanding of LA metabolism in the microbial cells, which is also important for the search for new antimicrobial drugs. This research is, therefore, an introduction to such further studies.

In-vitro activity of lipoic acid against Ureaplasma urealyticum and Ureaplasma parvum isolated from women with infections of the urogenital tract. A pilot study.

Several species of Ureaplasma bacteria are known to be present in the urogenital tract of humans, in both healthy individuals and symptomatic patients. These pathogens are associated with urogenital tract infections, infertility problems and spontaneous abortion in humans. The present study involved 77 strains of Ureaplasma species (Ureaplasma spp.), including 21 Ureaplasma urealyticum (U. urealyticum) strains and 56 Ureaplasma parvum (U. parvum) strains. Lipoic acid (LA) and its reduced form dihydrolipoic acid (DHLA) are synthesized in all prokaryotic and eukaryotic cells. Research of recent years increasingly points to therapeutic properties of exogenously supplemented LA. In our study, we examined for the first time the effect of LA on the bacteria multiplication and its bactericidal activity against U. urealyticum and U. parvum. The LA concentrations used were: 1200 µg/ml, 120 µg/ml, and 12 µg/ml. The titer for each strain of Ureaplasma spp. was estimated using the color changing units (CCU) assay. For CCU measurements, a series of 10-fold dilutions of each cell culture in 0.9% NaCl (titration) was prepared and 1 CCU/ml was defined as the highest dilution of cells at which color change was detected. The strongest bacteriostatic and bactericidal effect of LA was observed at a concentration of 1200 µg/ml. In contrast, at lower LA concentrations, stimulation of the bacteria multiplication was noted for 14% of the total number of strains tested. Taken together, the current data provide novel findings about potential beneficial antimicrobial effects of LA.

[Voice on the treatment of ureaplasmas infections]. / Glos w sprawie leczenia zakazen ureaplazmowych.

The aim of the study was to determine susceptibility of ureaplasmas for selected antimicrobials belonging to 3 groups, which are applied clinically. Study was conducted on 39 U. parvum and 26 U. urealyticum clinical isolates. Doxycyclin, azithromycin and levofloxacin were applied during the study. In case of azithromycin U. urealyticum strains were more susceptible; MIC90 = 1 microg/ml and MIC range for both species mean that only a few strains are resistant (only U. parvum). In case of doxycyclin U. parvum strains were more susceptible and MIC ranges mean that there are no resistant strains in both species. In case of levofloxacin wide MIC range with values signifying resistance was found. Doxycyclin is a good choice for empirical treatment of infections with ureaplasma aetiology. In case of macrolides higher dosage is indicated, because there is a risk of clinical failure at lower doses. Quinolones should not be used in empirical treatment of ureaplasma infections.

[May ureaplasmas in genital tract of HPV-positive women influence abnormal cytology of cervix?]. / Czy obecnosc ureaplazm w drogach rodnych kobiet zakazonych HPV wplywa na wystepowanie zmian cytologicznych w obrebie szyjki macicy?

The aim of the study was to estimate the incidence of Ureaplasma urealyticum (U.u.) and Ureaplasma parvum (Up.) in 168 women diagnosed with LSIL infected and not infected with HPV vs. 82 women with no cytological abnormalities in the cervix (control group). The material used in the study were cervical secretions samples. PCR was used to confirm the presence of HPV and to identify the species of ureaplasmas. U.p. was significantly more frequent in both groups of women. In the study group, ureaplasmas were more frequently isolated in the HPV infected (31%) vs. HPV negative (16%) women. No direct relationship was found between ureaplasmas and LSIL. Statistical analysis showed, that infection with HPV occurred more frequently in the presence of ureaplasmas (OR = 1.79; 95% PU 0.90-3.53; p = 0.093). The above relationship was most evident for U.u. The risk for HPV infection in that case was 6.5 fold higher. Infections with ureaplasmas, especially U.u should be considered as a factor increasing the risk of HPV infection of the cervical epithelial cells.

[Antimicrobial susceptibility of Ureaplasma urealyticum and ureaplasma parvum isolated from premature infants with respiratory disorders]. / Wrazliwosc na antybiotyki i chemioterapeutyki Ureaplasma urealyticum i Ureaplasma parvum izolowanych od noworodków z zaburzeniami oddechowymi.

UNLABELLED: The aim of the study was to analyse antimicrobial susceptibility of ureaplasmas isolated from the tracheal aspirates from premature infants with respiratory disorders. The study encompassed 73 ureaplasma strains, 47 belonging to U parvum (U.p.) species and 26 to U. urealyticum (U.u.). The strains were isolated paralelly on BioMerieux as well as liquid and solid PPLO media. Identification of studied strains was perfomed using PCR with primers specific to both ureaplasma species. Susceptibility to doxycycline (DOX), tetracyclin (TET), josamycin (JOS), azithromycin (AZY), erythromycin (ERY), clarytromycin (CLA), ofloxacin (OFL), ciprofloxacin (CIP), pristinamycin (PRI) was tested using a BioMerieux Mycoplasma IST 2 kit. RESULTS: in 7 of 9 examined antimicrobials the percentage of susceptible U.u. was lower thain the percentage of U.p. susceptible strains. CONCLUSIONS: The biggest differences related to susceptibility reffered to macrolides. Higher resistance of U.u. species to antimicrobials may suggest its higher pathogenecity.

Assessment of various diagnostic methods of ureaplasma respiratory tract infections in newborns.

We compared three methods used microbial culturing for detection of ureaplasmas in endotracheal aspirate from 500 prematurely born neonates with respiratory disturbances: BioMerieux test, PCR and microbial culturing. Ureaplasmas were detected in respiratory tracts of 79 (16%) newborns. Correlation of the results of culture with those obtained with the BioMerieux kit, culture with PCR and BioMerieux kit with PCR was 97%, 89% and 90%, respectively. Sensitivity and specificity of PCR in comparison with culture was 86% and 98%, respectively, and of the BioMerieux kit 96% and 98%. PCR can be recommended in rapid diagnostics of respiratory infections in newborns suffering from respiratory disorders. It allows the detection of ureaplasmas in case of parallel infections and identification of their species.

[Influence of ureaplasma infection on the clinical state of newborns]. / Wplyw zakazenia ureaplazmami na stan kliniczny noworodków.

UNLABELLED: The aim of this study was the analysis of the clinical state of newborns infected with various species of ureaplasma. METHODS: 50 prematurely born patients with respiratory disturbances and confirmed presence of ureaplasma in the respiratory tract were analyzed. Endotracheal aspirates were collected for examination. Presence of ureaplasma was confirmed by culture and a commercial test (Biomerieux), the ureaplasma species were identified using PCR. RESULTS: In 40 examined newborns Ureaplasma parvum (U.p.) was found, in 10 Ureaplasma urealyticum (U.u.). Newborns infected with U.u. were subject to more frequent and longer therapeutic procedures supporting respiration (respirator, nCPAP), needed more frequent surfactant and antibiotic administration. In the mentioned group the mortality rate was 33%, while in newborns infected with U.p. it was 15%. CONCLUSIONS: Initial results suggest worse clinical status and higher mortality of prematurely born infected with Ureaplasma urealyticum.