Modulation of LPS-induced chorioamnionitis by ureaplasma parvum in sheep chorioamnionitis in sheep

ABSTRACT Objective: Ureaplasma parvum colonization in the setting of polymicrobial flora is common in women with chorioamnionitis, and is a risk factor for preterm delivery and neonatal morbidity. We hypothesized that ureaplasma colonization of amniotic fluid will modulate chorioamnionitis induced by E.coli lipopolysaccharide (LPS). Methods: Sheep received intra-amniotic (IA) injections of media (control) or live ureaplasma either 7 or 70d before delivery. Another group received IA LPS 2d before delivery. To test for interactions, U.parvum exposed animals were challenged with IA LPS, and delivered 2d later. All animals were delivered preterm at 125±1 day gestation. Results: Both IA ureaplasmas and LPS induced leukocyte infiltration of chorioamnion. LPS greatly increased the expression of pro-inflammatory cytokines and myeloperoxidase in leukocytes, while ureaplasmas alone caused modest responses. Interestingly, 7d but not 70d ureaplasma exposure significantly downregulated LPS induced pro-inflammatory cytokines and myeloperoxidase expression in the chorioamnion. Conclusion: U.parvum can suppress LPS induced experimental chorioamnionitis.

Ureaplasma parvum multiple banded antigen (MBA) size variation - association with fetal inflammation in a sheep model (Published abstract)

Background: Ureaplasma species are the most prevalent isolates from women who deliver preterm. The MBA, a surface exposed lipoprotein, is a key virulence factor of ureaplasmas. We investigated MBA variation after chronic and acute intra-amniotic (IA) ureaplasma infections. Method: U. parvum serovar 3 (2x104 colony-forming-units) was injected IA into pregnant ewes at: 55 days gestation (d, term = 145d) (n=8); 117d (n=8) and 121d (n=8). Fetuses were delivered surgically (124d) and ureaplasmas cultured from amniotic fluid (AF), chorioamnion, fetal lung (FL) and umbilical cord were tested by western blot and PCR assays to demonstrate MBA and mba gene variation respectively. Tissue sections were sectioned and stained by haemotoxylin and eosin and inflammatory cell counts and pathology were reported (blinded to outcome). Results: Numerous MBA/mba variants were generated in vivo after chronic exposure to ureaplasma infection but after acute infection no variants (3d) or very few variants (7d) were generated. Identical MBA variants were detected within the AF and FL but different ureaplasma variants were detected within chorioamnion specimens. The severity of inflammation within chronically infected tissues varied between animals ranging from no inflammation to severe inflammation with/without fibrosis. Chorioamnion, FL and cord from the same animal demonstrated the same degree of inflammation. Conclusions: MBA/mba variation in vivo occurred after the initiation of the host immune response and we propose that ureaplasmas vary the MBA antigen to evade the host immune response. In some animals there was no inflammation despite colonisation with high numbers of ureaplasmas.

Genetic variability and antimicrobial resistance of Ureaplasma parvum in response to maternal erythromycin treatment : a study in pregnant sheep

Background: Ureaplasmas are the most frequently isolated microorganisms from the amniotic fluid (AF) of pregnant women and can cause chronic infections that are difficult to eradicate with standard macrolide treatment. We tested the effects of erythromycin treatment on phenotypic and genotypic markers of ureaplasmal antimicrobial resistance in sheep. Method: At 50 days of gestation (d, term=145d) 12 pregnant ewes received intra-amniotic injections of U. parvum serovar 3 (erythromycin-sensitive, 2x104 colony-forming-units). At 100d ewes received: erythromycin treatment (500 mg, q3h for 4 days, IM, n=6) or no treatment (n=6). Fetuses were delivered surgically (125d) and AF and chorioamnion were collected for: culture, minimum inhibitory concentration (MIC) and minimum biofilm inhibitory concentration (MBIC) testing; 23S rRNA sequencing; and detection of macrolide-lincosamide-streptogramin resistance (MLSr) genes. Results: MICs of erythromycin, azithromycin and roxithromycin against AF isolates were low (range = 0.06 mg/L to 1.0 mg/L); however, chorioamnion isolates demonstrated increased resistance to roxithromycin (0.13 – 5.33 mg/L). 62.5% of chorioamnion ureaplasmas formed biofilms in vitro and mutations (125 nucleotides, 29.6%) were found in the 23S rRNA gene (domain V) of chorioamnion (but not AF) ureaplasmas. MLSr genes (ermB, msrC and msrD) were detected in 100% of chorioamnion isolates and only msrD was detected in AF isolates (40%). Conclusions: 23S rRNA mutations and MLSr genes occurred independently of erythromycin treatment, suggesting that the anatomical site of infection and microenvironment may exert selective pressures on ureaplasmas that cause genetic changes and alter antimicrobial sensitivity profiles. These results have serious implications for treatment of in utero infections.

Ureaplasma parvum serovar 3 multiple banded antigen size variation after chronic intra-amniotic infection/colonization

Ureaplasma species are the microorganisms most frequently associated with adverse pregnancy outcomes. The multiple banded antigen (MBA), a surface-exposed lipoprotein, is a key virulence factor of ureaplasmas. The MBA demonstrates size variation, which we have shown previously to be correlated with the severity of chorioamnion inflammation. We aimed to investigate U. parvum serovar 3 pathogenesis in vivo, using a sheep model, by investigating: MBA variation after long term (chronic) and short term (acute) durations of in utero ureaplasma infections, and the severity of chorioamnionitis and inflammation in other fetal tissues. Inocula of 2x107 colony-forming-units (CFU) of U. parvum serovar 3 (Up) or media controls (C) were injected intra-amniotically into pregnant ewes at one of three time points: day 55 (69d Up, n=8; C69, n=4); day 117 (7d Up, n=8; C7, n=2); and day 121 (3d Up, n=8; C3, n=2) of gestation (term=145-150d). At day 124, preterm fetuses were delivered surgically. Samples of chorioamnion, fetal lung, and umbilical cord were: (i) snap frozen for subsequent ureaplasma culture, and (ii) fixed, embedded, sectioned and stained by haematoxylin and eosin stain for histological analysis. Selected fetal lung clinical ureaplasma isolates were cloned and filtered to obtain cultures from a single CFU. Passage 1 and clone 2 ureaplasma cultures were tested by western blot to demonstrate MBA variation. In acute durations of ureaplasma infection no MBA variants (3d Up) or very few MBA variants (7d Up) were present when compared to the original inoculum. However, numerous MBA size variants were generated in vivo (alike within contiguous tissues, amniotic fluid and fetal lung, but different variants were present within chorioamnion), during chronic, 69d exposure to ureaplasma infection. For the first time we have shown that the degree of ureaplasma MBA variation in vivo increased with the duration of gestation.

Developing schedules for mixed rail and road cane transport systems

This paper addresses the problem of scheduling a cane transport system involving both rail transport and road transport, where the road transport operates from several sidings in the rail network. An iterative approach for scheduling the rail transport system has been developed using existing rail transport scheduling tools. The assumption that harvesters serviced by road transport are effectively operating from the rail siding from which their bins are supplied seems a reasonable starting point for the analysis. There is a need to manually modify the schedule to take into account the road transport schedule to ensure that full bins are not collected before the road transport system delivers them back to the rail siding.

The power of art, the power of entertainment

The question of the relationship between culture and power continues to exercise researchers. In this paper I argue that it is useful to consider the differences between ‘art’ and ‘entertainment’ as systems of culture, each involving a distinct set of power relationships between producers and audiences. Art wants to change audiences; entertainment wants to be changed by audiences. From these different starting points a series of differences unfold in the power possessed by producers and audiences. Artists pride themselves in not involving the audience in the process of making art. By contrast, entertainment wants audiences to contribute to the making of texts. As to the question of who controls the range of what forms of culture are available, it seems that entertainment consumers – unlike art consumers – are ill-disciplined. Historical evidence demonstrates that if legal corporate providers do not offer the kinds of entertainment they want, they will turn to illegal sources. The different ways in which ‘art’ and ‘entertainment’ function as cultural systems suggest that we must rethink our positions on ‘media power’.

Methodological issues in defining aggression for content analyses of sexually explicit material

There exists an important tradition of content analyses of aggression in sexually explicit material. The majority of these analyses use a definition of aggression that excludes consent. This article identifies three problems with this approach. First, it does not distinguish between aggression and some positive acts. Second, it excludes a key element of healthy sexuality. Third, it can lead to heteronormative definitions of healthy sexuality. It would be better to use a definition of aggression such as Baron and Richardson's (1994) in our content analyses, that includes a consideration of consent. A number of difficulties have been identified with attending to consent but this article offers solutions to each of these.

Pro-Am curators of Australian television history : how is their practice different from that of professional television historians?

Eleven Pro-Am curators of Australian television history were interviewed about their practice. The data helps us to understand the relationship between professional and Pro-Am approaches to Australian television history. There is no simple binary – the lines are blurred – but there are some differences. Pro-Am curators of Australian television history are not paid for their work and present other motivations for practice – particularly being that ‘weird child’ who was obsessed with gathering information and objects related to television. They have freedom to curate only programs and genres that interest them, and they tend to collect merchandise as much as program texts themselves. And they have less interest in formally cataloguing their material than do professional curators of Australian television history.

Why are children the most important audience for pornography in Australia?

Who watches pornography in Australia? If you listen to public debates about the genre the answer is clear – it’s children. Children are accessing pornography on smartphones (Murray and Tin 2011). Children are taking ‘lewd’ photographs of themselves, creating their own pornography (Nelligan and Etheridge 2011). Indigenous Australian children must be protected by banning pornography (the Age 2011). Pornographic magazines are placed where children can see them (O'Rourke 2011). Exposure to pornography is damaging children (Sundstrom 2011). The Australian Government insists that the Internet must be filtered to protect children from pornography (Collerton 2010). And if indeed any adults are watching pornography in Australia, then it’s child pornography (MacDonald 2011; Ralston and Howden 2011).In story after story, public debate about pornography focuses on children as its audience. There is no suggestion that children are numerically the largest audience of pornography in Australia. But emphatically the suggestion is that children are the most important audience to be taken into account when thinking about the genre. This chapter explores why this is the case, and notes the political advantages and disadvantages of focusing on children as the most important audience for pornography in Australia.

Masturbation and the media

This paper explores the important role the media play in informing young people about masturbation. A pilot study of focus groups with twenty-two young Australians, aged between 14-16 explored what they know about sex and sexuality, and where they have found that knowledge. This paper reports on their knowledge about masturbation. Although researchers agree that masturbation can be a positive part of healthy sexual development, most young people reported that they received very little positive information about it from their parents or in formal sex education in school. These young people’s discussions around this topic were largely ambivalent, but also highly complex due to the varying levels and types of information that they receive. In this context the media play a vital role in providing information about masturbation through books and magazines for young women, and television comedies for young men.

Speaking truth to power : sexism, outrage and the public consciousness

Alan Jones's now infamous comment that women are destroying the joint' sparked a flurry of observations and interpretations in the national and international media and in the blogosphere about the motivations and meanings of such a statement. The overriding tone of this commentary was outrage - how dare an influential figure like Jones state with such blatant and provocative hostility that women are at best unsuitable for leadership and at worst, damaging our public and private institutions? The answers, as we have seen, are likely to be complex, but an alternative question could also be posed. Why the outrage? Public male figures have, throughout history, made derogatory, demeaning and sexist remarks about women, and the material effects of gender inequality continue to be felt- consider the gender pay gap and glass ceiling phenomenon for starters. So why did this particular comment ignite such widespread indignation and anger?

Effects of chronic intra-amniotic Ureaplasma parvum (serovar 3 and 6) colonization in the ovine fetus

Background: We have developed a sheep model of intrauterine ureaplasma infection. We aimed to examine the capability of ureaplasmas in the amniotic fluid to infect the fetus and alter fetal development...