Microbial colonisation of human follicular fluid and adverse in vitro fertilisation outcomes

This study, investigating 263 women undergoing trans-vaginal oocyte retrieval for in vitro fertilisation (IVF) found that microorganisms colonising follicular fluid contributed to adverse IVF (pre-implantation) and pregnancy (post-implantation) outcomes including poor quality embryos, failed pregnancy and early pregnancy loss (< 37 weeks gestation). Some microorganisms also showed in vitro growth patterns in liquid media that appeared to be enhanced by the hormonal stimulation protocol used for oocyte retrieval. Elaborated cytokines within follicular fluid were also associated with adverse IVF outcomes. This study is imperative because infertility affects 16% of the human population and the numbers of couples needing assistance continues to increase. Despite significant improvements in the technical aspects of assisted reproductive technologies (ART), the live birth rate has not increased proportionally. Overt genital tract infection has been associated with both infertility and adverse pregnancy outcomes (including miscarriage and preterm birth) as a direct result of the infection or the host response to it. Importantly, once inflammation had become established, medical treatment often failed to prevent these significant adverse outcomes. Current evaluations of fertility focus on the ovary as a site of steroid hormone production and ovulation. However, infertility as a result of subclinical colonisation of the ovary has not been reported. Furthermore, identification of the microorganisms present in follicular fluid and the local cytokine profile may provide clinicians with an early indication of the prognosis for IVF treatment in infertile couples, thus allowing antimicrobial treatment and/or counselling about possible IVF failure. During an IVF cycle, multiple oocytes undergo maturation in vivo in response to hormonal hyperstimulation. Oocytes for in vitro insemination are collected trans-vaginally. The follicular fluid that bathes the maturing oocyte in vivo, usually is discarded as part of the IVF procedure, but provides a unique opportunity to investigate microbial causes of adverse IVF outcomes. Some previous studies have identified follicular fluid markers that predict IVF pregnancy outcomes. However, there have not been any detailed microbiological studies of follicular fluid. For this current study, paired follicular fluid and vaginal secretion samples were collected from women undergoing IVF cycles to determine whether microorganisms in follicular fluid were associated with adverse IVF outcomes. Microorganisms in follicular fluid were regarded as either "colonisers" or "contaminants"; colonisers, if they were unique to the follicular fluid sample, and contaminants if the same microorganisms were detected in the vaginal and follicular fluid samples indicating that the follicular fluid was merely contaminated during the oocyte retrieval process. Quite unexpectedly, by these criteria, we found that follicular fluid from approximately 30% of all subjects was colonised with bacteria. Fertile and infertile women with colonised follicular fluid had decreased embryo transfer rates and decreased pregnancy rates compared to women with contaminated follicular fluids. The observation that follicular fluid was not always sterile, but contained a diverse range of microorganisms, is novel. Many of the microorganisms we detected in follicular fluid are known opportunistic pathogens that have been detected in upper genital tract infections and are associated with adverse pregnancy outcomes. Bacteria were able to survive for at least 28 weeks in vitro, in cultures of follicular fluid. Within 10 days of establishing these in vitro cultures, several species (Lactobacillus spp., Bifidobacterium spp., Propionibacterium spp., Streptococcus spp. and Salmonella entericus) had formed biofilms. Biofilms play a major role in microbial pathogenicity and persistence. The propensity of microbial species to form biofilms in follicular fluid suggests that successful treatment of these infections with antimicrobials may be difficult. Bifidobacterium spp. grew, in liquid media, only if concentrations of oestradiol and progesterone were similar to those achieved in vivo during an IVF cycle. In contrast, the growth of Streptococcus agalactiae and Escherichia coli was inhibited or abolished by the addition of these hormones to culture medium. These data suggest that the likelihood of microorganisms colonising follicular fluid and the species of bacteria involved is influenced by the stage of the menstrual cycle and, in the case of IVF, the nature and dose of steroid hormones administered for the maturation of multiple oocytes in vivo. Our findings indicate that the elevated levels of steroid hormones during an IVF cycle may influence the microbial growth within follicular fluid, suggesting that the treatment itself will impact on the microflora present in the female upper genital tract during pre-conception and early post-conception phases of the cycle. The effect of the host immune response on colonising bacteria and on the outcomes of IVF also was investigated. White blood cells reportedly compose between 5% and 15% of the cell population in follicular fluid. The follicular membrane is semi-permeable and cells are actively recruited as part of the normal menstrual cycle and in response to microorganisms. A previous study investigated follicular fluid cytokines from infertile women and fertile oocyte donors undergoing IVF, and concluded that there were no significant differences in the cytokine concentrations between the two groups. However, other studies have reported differences in the follicular fluid cytokine levels associated with infertile women with endometriosis or polycystic ovary syndrome. In this study, elevated levels of interleukin (IL)-1 á, IL-1 â and vascular endothelial growth factor (VEGF) in vaginal fluid were associated with successful fertilisation, which may be useful marker for successful fertilisation outcomes for women trying to conceive naturally or prior to oocyte retrieval for IVF. Elevated levels of IL-6, IL-12p40, granulocyte colony stimulating factor (GCSF) and interferon-gamma (IFN ã) in follicular fluid were associated with successful embryo transfer. Elevated levels of pro-inflammatory IL-18 and decreased levels of anti-inflammatory IL-10 were identified in follicular fluid from women with idiopathic infertility. Successful fertilisation and implantation is dependent on a controlled pro-inflammatory environment, involving active recruitment of pro-inflammatory mediators to the genital tract as part of the menstrual cycle and early pregnancy. However, ongoing pregnancy requires an enhanced anti-inflammatory environment to ensure that the maternal immune system does not reject the semi-allergenic foetus. The pro-inflammatory skew in the follicular fluid of women with idiopathic infertility, correlates with normal rates of fertilisation, embryo discard and embryo transfer, observed for this cohort, which were similar to the outcomes observed for fertile women. However, their pregnancy rate was reduced compared to fertile women. An altered local immune response in follicular fluid may provide a means of explaining infertility in this cohort, previously defined as 'idiopathic'. This study has found that microorganisms colonising follicular fluid may have contributed to adverse IVF and pregnancy outcomes. Follicular fluid bathes the cumulus oocyte complex during the in vivo maturation process, and microorganisms in the fluid, their metabolic products or the local immune response to these microorganisms may result in damage to the oocytes, degradation of the cumulus or contamination of the IVF culture system. Previous studies that have discounted bacterial contamination of follicular fluid as a cause of adverse IVF outcomes failed to distinguish between bacteria that were introduced into the follicular fluid at the time of trans-vaginal oocyte retrieval and those that colonised the follicular fluid. Those bacteria that had colonised the fluid may have had time to form biofilms and to elicit a local immune response. Failure to draw this distinction has previously prevented consideration of bacterial colonisation of follicular fluid as a cause of adverse IVF outcomes. Several observations arising from this study are of significance to IVF programs. Follicular fluid is not always sterile and colonisation of follicular fluid is a cause of adverse IVF and pregnancy outcomes. Hormonal stimulation associated with IVF may influence whether follicular fluid is colonised and enhance the growth of specific species of bacteria within follicular fluid. Bacteria in follicular fluid may form biofilms and literature has reported that this may influence their susceptibility to antibiotics. Monitoring the levels of selected cytokines within vaginal secretions may inform fertilisation outcomes. This study has identified novel factors contributing to adverse IVF outcomes and that are most likely to affect also natural conception outcomes. Early intervention, possibly using antimicrobial or immunological therapies may reduce the need for ART and improve reproductive health outcomes for all women.

The stimulation of proliferation and differentiation of periodontal ligament cells by the ionic products from Ca7Si2P2O16 bioceramics

The ultimate goal of periodontal tissue engineering is to produce predictable regeneration of alveolar bone, root cementum, and periodontal ligament, which are lost as a result of periodontal diseases. To achieve this goal, it is of great importance to develop novel bioactive materials which could stimulate the proliferation, differentiation and osteogenic/cementogenic gene expression of periodontal ligament cells (PDLCs) for periodontal regeneration. In this study, we synthesized novel Ca7Si2P2O16 ceramic powders for the first time by the sol–gel method and investigated the biological performance of PDLCs after exposure to different concentrations of Ca7Si2P2O16 extracts. The original extracts were prepared at 200 mg ml-1 and further diluted with serum-free cell culture medium to obtain a series of diluted extracts (100, 50, 25, 12.5 and 6.25 mg ml–1). Proliferation, alkaline phosphatase(ALP) activity, Ca deposition, and osteogenesis/cementogenesis-related gene expression (ALP, Col I, Runx2 and CEMP1) were assayed for PDLCs on days 7 and 14. The results showed that the ionic products from Ca7Si2P2O16 powders significantly stimulated the proliferation, ALP activity, Ca deposition and osteogenesis/cementogenesisrelated gene expression of PDLCs. In addition, it was found that Ca7Si2P2O16 powders had excellent apatite-mineralization ability in simulated body fluids. This study demonstrated that Ca7Si2P2O16 powders with such a specific composition possess the ability to stimulate the PDLC proliferation and osteoblast/cemenoblast-like cell differentiation, indicating that they are a promising bioactive material for periodontal tissue regeneration application.

Questioning pedagogies : Hong Kong pre-service teachers' dialogic reflections on a transnational school experience

There are increasing opportunities in many countries for pre-service teachers to engage in a transnational school-based experience as part of study abroad programmes. The transformative potential of such transnational teaching experiences is recorded in research studies, often supported by data from participant surveys. However, there has been a lack of evidence investigating how shifts in professional understanding derive from such experiences. This qualitative study addresses this issue by exploring the perspectives of 16 pre-service teachers of English as a Second language from Hong Kong, who engaged in transnational teaching activities with primary school pupils in Australia, during their study abroad program. Discourse analysis of participants’ dialogues traces how they encountered conflicting Discourses of ‘student-centredness’ in the Australian classroom. Reflecting dialogically on their experiences led participants to negotiate and reframe their understandings of language teaching pedagogy and themselves as language teachers. The findings demonstrate the importance of both peer and lecturer feedback into the process of dialogic reflection and the need for more longitudinal research into the impact of transnational school-based experience in pre-service teacher education.

New maps of learning for quality art education : what pre-service teachers should learn and be able to do

It would be a rare thing to visit an early years setting or classroom in Australia that does not display examples of young children’s artworks. This practice serves to give schools a particular ‘look’, but is no guarantee of quality art education. The Australian National Review of Visual Arts Education (NRVE) (2009) has called for changes to visual art education in schools. The planned new National Curriculum includes the arts (music, dance, drama, media and visual arts) as one of the five learning areas. Research shows that it is the classroom teacher that makes the difference, and teacher education has a large part to play in reforms to art education. This paper provides an account of one foundation unit of study (Unit 1) for first year university students enrolled in a 4-year Bachelor degree program who are preparing to teach in the early years (0–8 years). To prepare pre-service teachers to meet the needs of children in the 21st century, Unit 1 blends old and new ways of seeing art, child and pedagogy. Claims for the effectiveness of this model are supported with evidence-based research, conducted over the six years of iterations and ongoing development of Unit 1.

Assessing for work integrated learning experiences : a pre-service teacher perspective

A critical aspect of the debate about work integrated learning in the university context is the blurring of boundaries and responsibilities in terms of student learning. In an Australian pre-service teacher education program this blurring of boundaries is apparent in stakeholder tensions about the nature and role of assessment during the practicum. In the study reported in this paper, students responded positively to the content of assessment tasks but maintained that their efforts to implement the associated planning in the workplace were stymied because of disparate understandings between university and school staff about the purpose of the task.

UK pre-pack reforms : pause for thought in Australia?

On 31 March 2011 the UK Government announced new measures to regulate the use of pre-packaged sales in administration. The legislation is not expected until later in 2011, but the announcement heralds a shift in regulatory attitudes towards pre-packs in the UK which should give all local pre-pack advocates pause for thought when considering the merits of embracing the procedure in Australia. In the Jan-March 2011 edition of the Australian Insolvency Journal, an interesting article by Nicholas Crouch and Shabnam Amirbeaggi extolled the virtues of pre-packs and called for “legislative reform to embrace pre-packs” in Australia. By way of reply (and in a spirit of constructive debate) this article respectfully contends that while pre-packs certainly have their place in preserving business value in certain circumstances, Australia should be careful not to sleepwalk into adopting a procedure which legitimises phoenixing at the expense of creditor confidence and participation in our insolvency regime.

Dealing with unfinished business : preparing pre-service social education teachers for curriculum change

Pre-service teacher education is unfinished business. New social education teachers face the challenge of fluid policy environments in which curriculum content and pedagogy are continually changing. The evolving Australian curriculum is the most recent example of such fluidity with its emphasis on shifting the educational agenda to a focus on discipline-based approaches. This paper addresses the concerns of final year pre-service and early career social education teachers, in terms of their professional development needs, by drawing on the findings of a pilot study with students and recent graduates from a university in south-east Queensland. It concludes that social education curriculum units which embed links to professional practice and professional development in teaching, learning and assessment may provide the way forward for enhancing the transition to practice for beginning teachers and assist them in navigating constant change.

Dynamic compression improves biosynthesis of human zonal chondrocytes from osteoarthritis patients

Objective: We hypothesize that chondrocytes from distinct zones of articular cartilage respond differently to compressive loading, and that zonal chondrocytes from osteoarthritis (OA) patients can benefit from optimized compressive stimulation. Therefore, we aimed to determine the transcriptional response of superficial (S) and middle/deep (MD) zone chondrocytes to varying dynamic compressive strain and loading duration. To confirm effects of compressive stimulation on overall matrix production, we subjected zonal chondrocytes to compression for 2 weeks. Design: Human S and MD chondrocytes from osteoarthritic joints were encapsulated in 2% alginate, pre-cultured, and subjected to compression with varying dynamic strain (5, 15, 50% at 1 Hz) and loading duration (1, 3, 12 h). Temporal changes in cartilage-specific, zonal, and dedifferentiation genes following compression were evaluated using quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR). The benefits of long-term compression (50% strain, 3 h/day, for 2 weeks) were assessed by measuring construct glycosaminoglycan (GAG) content and compressive moduli, as well as immunostaining. Results: Compressive stimulation significantly induced aggrecan (ACAN), COL2A1, COL1A1, proteoglycan 4 (PRG4), and COL10A1 gene expression after 2 h of unloading, in a zone-dependent manner (P < 0.05). ACAN and PRG4 mRNA levels depended on strain and load duration, with 50% and 3 h loading resulting in highest levels (P < 0.05). Long-term compression increased collagen type II and ACAN immunostaining and total GAG (P < 0.05), but only S constructs showed more PRG4 stain, retained more GAG (P < 0.01), and developed higher compressive moduli than non-loaded controls. Conclusions: The biosynthetic activity of zonal chondrocytes from osteoarthritis joints can be enhanced with selected compression regimes, indicating the potential for cartilage tissue engineering applications. © 2012 Osteoarthritis Research Society International.

The heart of quality teaching : a values-based pedagogy for pre-service teacher education

In an ever changing world the adults of the future will be faced with many challenges. To cope with these challenges it seems apparent that values education will need to become paramount within a child.s education. A considerable number of research studies have indicated that values education is a critical component within education (Lovat & Toomey, 2007b). Building on this research Lovat (2006) claimed that values education was the missing link in quality teaching The concept of quality teaching had risen to the fore within educational research literature in the late 20th century with the claim that it is the teacher who makes the difference in schooling (Hattie, 2004). Thus, if teachers make such a difference to student learning, achievement and well-being, then it must hold true that pre-service teacher education programmes are vital in ensuring the development of quality teachers for our schools. The gap that this current research programme addressed was to link the fields of values education, quality teaching and pre-service teacher education. This research programme aimed to determine the impact of a values-based pedagogy on the development of quality teaching dimensions within pre-service teacher education. The values-based pedagogy that was investigated in this research programme was Philosophy in the Classroom. The research programme adopted a nested case study design based on the constructivist-interpretative paradigm in examining a unit within a pre-service teacher education programme at a Queensland university. The methodology utilised was qualitative where the main source of data was via interviews. In total, 43 pre-service teachers participated in three studies in order to determine if their involvement in a unit where the focus was on introducing pre-service teachers to an explicit values-based pedagogy impacted on their knowledge, skills and confidence in terms of quality teaching dimensions. The research programme was divided into three separate studies in order to address the two research questions: 1. In what ways do pre-service teachers perceive they are being prepared to become quality teachers? 2. Is there a connection between an explicit values-based pedagogy in pre-service teacher education and the development of pre-service teachers. understanding of quality teaching? Study One provided insight into 21 pre-service teachers. understandings of quality teaching. These 21 participants had not engaged in an explicit values-based pedagogy. Study Two involved the interviewing of 22 pre-service teachers at two separate points in time . prior to exposure to a unit that employed a values-explicit pedagogy and post this subject.s lecture content delivery. Study Three reported on and analysed individual case studies of five pre-service teachers who had participated in Study Two Time 1 and Time 2, as well as a third time following their field experience where they had practice in teaching the values explicit pedagogy. The results of the research demonstrate that an explicit values-based pedagogy introduced into a teacher education programme has a positive impact on the development of pre-service teachers. understanding of quality teaching skills and knowledge. The teaching and practice of a values-based pedagogy positively impacted on pre-service teachers with increases of knowledge, skills and confidence demonstrated on the quality teaching dimensions of intellectual quality, a supportive classroom environment, recognition of difference, connectedness and values. These findings were reinforced through the comparison of pre-service teachers who had participated in the explicit values-based pedagogical approach, with a sample of pre-service teachers who had not engaged in this same values-based pedagogical approach. A solid values-based pedagogy and practice can and does enhance pre-service teachers. understanding of quality teaching. These findings surrounding the use of a values-based pedagogy in pre-service teacher education to enhance quality teaching knowledge and skills has contributed theoretically to the field of educational research, as well having practical implications for teacher education institutions and teacher educators.

District Court of Queensland Act 1968 s 68 : jurisdiction in pre-proceedings applications under Personal Injuries Proceedings Act 2002

In Woolworths Ltd v Graham [2007] QDC 301 Searles DCJ struck out a pre-proceedings application under the Personal Injuries Proceedings Act 2002 (Qld)on the basis that the material before the Court was not sufficient to attract the jurisdiction of the District Court.The decision serves more broadly as a reminder that the District Court is an inferior court of defined and limited jurisdiction and that any proceedings brought in it must be demonstrably within the jurisdiction conferred on that court by legislation.

Pre-sentence Report, Magisterial Discourse and Agency in Youth Courts in England

This article draws on interviews with Youth Court magistrates to examine if and how discourses, strategies and technologies of risk governance have affected Youth Court magistrates in England and Wales. The aim of the article is to detail the complex relationship between magisterial agency in decision making and youth justice policies which focus on risk control and management. The article demonstrates that, contrary to what might be assumed from the youth and risk governance theoretical literature, Youth Offending Team risk assessments form only one part of the information used by magistrates to explain young people’s presence in courts. This article concludes that magisterial decision making is framed not by formal, expert assessments of risk, but by magistrates’ claims that they are ‘knowing outsiders’, who through judicious use of information presented to them and their own life experiences are able to make objective judgements about both the risk assessments authored by Youth Offending Teams and the young lawbreakers before them.

Ethnic differences in body fat distribution among Asian pre-pubertal children: A cross-sectional multicenter study

Background Ethnic differences in body fat distribution contribute to ethnic differences in cardiovascular morbidities and diabetes. However few data are available on differences in fat distribution in Asian children from various backgrounds. Therefore, the current study aimed to explore ethnic differences in body fat distribution among Asian children from four countries. Methods A total of 758 children aged 8-10 y from China, Lebanon, Malaysia and Thailand were recruited using a non-random purposive sampling approach to enrol children encompassing a wide BMI range. Height, weight, waist circumference (WC), fat mass (FM, derived from total body water [TBW] estimation using the deuterium dilution technique) and skinfold thickness (SFT) at biceps, triceps, subscapular, supraspinale and medial calf were collected. Results After controlling for height and weight, Chinese and Thai children had a significantly higher WC than their Lebanese and Malay counterparts. Chinese and Thais tended to have higher trunk fat deposits than Lebanese and Malays reflected in trunk SFT, trunk/upper extremity ratio or supraspinale/upper extremity ratio after adjustment for age and total body fat. The subscapular/supraspinale skinfold ratio was lower in Chinese and Thais compared with Lebanese and Malays after correcting for trunk SFT. Conclusions Asian pre-pubertal children from different origins vary in body fat distribution. These results indicate the importance of population-specific WC cut-off points or other fat distribution indices to identify the population at risk of obesity-related health problems.