DNA technology for the detection of common genetic variants that predispose to thrombophilia

With the identification of common single locus point mutations as risk factors for thrombophilia, many DNA testing methodologies have been described for detecting these variations. Traditionally, functional or immunological testing methods have been used to investigate quantitative anticoagulant deficiencies. However, with the emergence of the genetic variations, factor V Leiden, prothrombin 20210 and, to a lesser extent, the methylene tetrahydrofolate reductase (MTHFR677) and factor V HR2 haplotype, traditional testing methodologies have proved to be less useful and instead DNA technology is more commonly employed in diagnostics. This review considers many of the DNA techniques that have proved to be useful in the detection of common genetic variants that predispose to thrombophilia. Techniques involving gel analysis are used to detect the presence or absence of restriction sites, electrophoretic mobility shifts, as in single strand conformation polymorphism or denaturing gradient gel electrophoresis, and product formation in allele-specific amplification. Such techniques may be sensitive, but are unwielding and often need to be validated objectively. In order to overcome some of the limitations of gel analysis, especially when dealing with larger sample numbers, many alternative detection formats, such as closed tube systems, microplates and microarrays (minisequencing, real-time polymerase chain reaction, and oligonucleotide ligation assays) have been developed. In addition, many of the emerging technologies take advantage of colourimetric or fluorescence detection (including energy transfer) that allows qualitative and quantitative interpretation of results. With the large variety of DNA technologies available, the choice of methodology will depend on several factors including cost and the need for speed, simplicity and robustness. © 2000 Lippincott Williams & Wilkins.

A conceptual decision-making framework for the delivery of innovative change in organisations

Research found that today’s organisations are increasingly aware of the potential barriers and perceived challenges associated with the successful delivery of change — including cultural and sub-cultural indifferences; financial constraints; restricted timelines; insufficient senior management support; fragmented key stakeholder commitment; and inadequate training. The delivery and application of Innovative Change (see glossary) within a construction industry organisation tends to require a certain level of ‘readiness’. This readiness is the combination of an organisation’s ability to part from undertakings that may be old, traditional, or inefficient; and then being able to readily adopt a procedure or initiative which is new, improved, or more efficient. Despite the construction industry’s awareness of the various threats and opportunities associated with the delivery of change, research found little attention is currently given to develop a ‘decision-making framework’ that comprises measurable elements (dynamics) that may assist in more accurately determining an organisation’s level of readiness or ability to deliver innovative change. To resolve this, an initial Background Literature Review in 2004 identified six such dynamics, those of Change, Innovation, Implementation, Culture, Leadership, and Training and Education, which were then hypothesised to be key components of a ‘Conceptual Decision-making Framework’ (CDF) for delivering innovative change within an organisation. To support this hypothesis, a second (more extensive) Literature Review was undertaken from late 2007 to mid 2009. A Delphi study was embarked on in June 2008, inviting fifteen building and construction industry members to form a panel and take part in a Delphi study. The selection criterion required panel members to have senior positions (manager and above) within a recognised field or occupation, and to have experience, understanding and / or knowledge in the process of delivering change within organisations. The final panel comprised nine representatives from private and public industry organisations and tertiary / research and development (R&D) universities. The Delphi study developed, distributed and collated two rounds of survey questionnaires over a four-month period, comprising open-ended and closed questions (referred to as factors). The first round of Delphi survey questionnaires were distributed to the panel in August 2008, asking them to rate the relevancy of the six hypothesised dynamics. In early September 2008, round-one responses were returned, analysed and documented. From this, an additional three dynamics were identified and confirmed by the panel as being highly relevant during the decision-making process when delivering innovative change within an organisation. The additional dynamics (‘Knowledge-sharing and Management’; ‘Business Process Requirements’; and ‘Life-cycle Costs’) were then added to the first six dynamics and used to populate the second (final) Delphi survey questionnaire. This was distributed to the same nine panel members in October 2008, this time asking them to rate the relevancy of all nine dynamics. In November 2008, round-two responses were returned, analysed, summarised and documented. Final results confirmed stability in responses and met Delphi study guidelines. The final contribution is twofold. Firstly, findings confirm all nine dynamics as key components of the proposed CDF for delivering innovative change within an organisation. Secondly, the future development and testing of an ‘Innovative Change Delivery Process’ (ICDP) is proposed, one that is underpinned by an ‘Innovative Change Decision-making Framework’ (ICDF), an ‘Innovative Change Delivery Analysis’ (ICDA) program, and an ‘Innovative Change Delivery Guide’ (ICDG).

Partial protection against chlamydial reproductive tract infection by a recombinant major outer membrane protein/CpG/cholera toxin intranasal vaccine in the guinea pig Chlamydia cavaie model

Chlamydia trachomatis is a major cause of sexually transmitted diseases worldwide. There currently is no vaccine to protect against chlamydial infection of the female reproductive tract. Vaccine development has predominantly involved using the murine model, however infection of female guinea pigs with Chlamydia caviae more closely resembles chlamydial infection of the human female reproductive tract, and presents a better model to assess potential human chlamydial vaccines. We immunised female guinea pigs intranasally with recombinant major outer membrane protein (r-MOMP) combined with CpG-10109 and cholera toxin adjuvants. Both systemic and mucosal immune responses were elicited in immunised animals. MOMP-specific IgG and IgA were present in the vaginal mucosae, and high levels of MOMP-specific IgG were detected in the serum of immunised animals. Antibodies from the vaginal mucosae were also shown to be capable of neutralising C. caviae in vitro. Following immunisation, animals were challenged intravaginally with a live C. caviae infection of 102 inclusion forming units. We observed a decrease in duration of infection and a significant (p<0.025) reduction in infection load in r-MOMP immunised animals, compared to animals immunised with adjuvant only. Importantly, we also observed a marked reduction in upper reproductive tract (URT) pathology in r-MOMP immunised animals. Intranasal immunisation of female guinea pigs with r-MOMP was able to provide partial protection against C. caviae infection, not only by reducing chlamydial burden but also URT pathology. This data demonstrates the value of using the guinea pig model to evaluate potential chlamydial vaccines for protection against infection and disease pathology caused by C. trachomatis in the female reproductive tract.

A comparison of the effects of a chlamydial vaccine administered during or after a C. muridarum urogenital infection of female mice

There are approximately 92 million new chlamydial infections of the genital tract in humans diagnosed each year, costing health care systems billions of dollars in treatment not only of acute infections, but also of associated inflammatory sequelae, such as pelvic inflammatory disease (PID) and ectopic pregnancy. These numbers are increasing at a steady rate and, due to the asymptomatic nature of infections, the incidence may be underestimated and the costs of treatment therefore higher. Over the previous few decades there has been a large amount of research into the development of an efficacious vaccine against genital tract chlamydial infections. The majority of this research has focused on females, due to the high rate of development of associated diseases, including PID, which can lead to ectopic pregnancy and infertility. In light of the increasing infection rates that have occurred despite the availability of antibiotics, and the asymptomatic nature of chlamydial infections, it is imperative that an efficacious vaccine that protects against infection and associated pathology be developed.