The impact of three-dimensional visualisation on midwifery students' application of knowledge of the third stage of labour to practice: Qualitative findings of a pilot randomised controlled trial.

PROBLEM: Complex physiological processes are often difficult for midwifery students to comprehend when using traditional teaching and learning approaches. Online resources for midwifery education are limited. BACKGROUND: Face to face instructional workshops using simulation have had some impact on improving understanding. However, in the 21st century new technologies offer the opportunity to provide alternative learning approaches. Virtual and artificial realities have been shown to increase confidence in decision making during clinical practice. AIM: Explore the impact of using three-dimensional (3D) visualisation in midwifery education, on student's application, when educating women about the birth of the placenta, and membranes. METHODS: Face to face individual interviews were performed, to collect deep, meaningful experiences of students, learning about the third stage of labour. FINDINGS: Prior clinical experiences impacted on student's ability to articulate how they would discuss birth of the placenta and membranes, and the process of haemostasis with women. DISCUSSION: The narrative findings of this pilot study identified ways that students traditionally learn midwifery, through theory, and clinical practice. Interview narratives illustrated how midwifery students who had previous experiences of witnessing birth, had superior ability to discuss the third stage of labour with women. While students with limited birth experiences, found the 3DMVR assisted them in their understanding of the physiology of the third stage of labour. CONCLUSION: In an environment of increasing technological advances, clinical placements remain an essential component of midwifery education.

The impact of three-dimensional visualisation on midwifery student learning, compared with traditional education for teaching the third stage of labour: A pilot randomised controlled trial.

BACKGROUND: Complex physiological processes are often difficult for midwifery students to comprehend when using traditional teaching and learning approaches. Face to face instructional workshops using simulation have had some impact on improving understanding. However, in the 21st century new technologies offer the opportunity to provide alternative learning approaches. AIM: To investigate the impact of using three-dimensional (3D) visualisation in midwifery education on student's experience of learning, and retention of knowledge at three points in time. DESIGN: A pilot study involving a two-armed parallel Randomised Controlled Trial (RCT) comparing the retention of knowledge scores between the control and intervention groups. SETTING: An Australian University in the Northern Territory. PARTICIPANTS: The sample included second year Bachelor of Midwifery students (n = 38). All received traditional midwifery education before being randomly allocated to either the intervention (n = 20) or control (n = 18) group. METHODS: A new immersive virtual environment was introduced to complement existing traditional midwifery education on the third stage of labour. This intervention was evaluated using a demographic survey and multiple-choice questionnaire to collect baseline information via Qualtrics. To measure change in knowledge and comprehension, participants completed the same multiple-choice knowledge questionnaire at three time points; pre, immediately post and at 1 month post intervention. In addition, the intervention group completed a 3D student satisfaction survey. RESULTS: Baseline knowledge scores were similar between the groups. A statistically significant increase in knowledge score was evident immediately post intervention for the intervention group, however there was no significant difference in knowledge score at one month. CONCLUSIONS: The results support the creation of further three-dimensional visualisation teaching resources for midwifery education. However, a larger randomised controlled study is needed to seek generalisation of these findings to confirm enhanced student learning and retention of knowledge post 3DMVR, beyond the immediate exposure time.

Implementation of distributed automated medication dispensing units in a new hospital: Nursing and pharmacy experience.

AIMS AND OBJECTIVES: To explore the structures, processes and outcomes involved in an Automated Medication Dispensing system implementation and its impact on patient safety. BACKGROUND: Increasing digitalisation of medication prescribing, dispensing, administration and stock management has occurred over the past two decades. While automated medication dispensing units aim to provide safe, high-quality, patient-centred care, the implementation may result in unintended consequences leading to suboptimal outcomes. DESIGN: This study uses a qualitative approach guided by Donabedian's structure, process and outcome framework. METHODS: Twenty-six registered nurses and pharmacy assistant staff, from clinical areas equipped with automated medication dispensing cabinets, participated in semi-structured interviews. In-depth, thematic analysis explored the structures and processes. Together with interview data, content analysis of text data generated by internal risk management and critical incident reporting systems was undertaken to evaluate outcomes. Findings were considered in light of the Interactive Sociotechnical Analysis approach to health information technology. The COREQ checklist was used in preparation of this article. RESULTS: Pharmacy assistants reported better satisfaction with the system at implementation than nurses. Training provided for nurses and their involvement in system implementation was reported as insufficient; however, nurses' use of and satisfaction with the system improved over time. A recursive relationship between the changes imposed by the system and nurses' creative problem solving (workarounds) used to manage these changes, impacted work productivity for nurses and safety for patients. CONCLUSIONS: The individualised nature of "workarounds" employed offered both risks and opportunities which require further identification, investigation and management. RELEVANCE TO CLINICAL PRACTICE: Nurses are the majority of the health workforce. Digitalisation of traditionally paper-based activities in health care, impacting nursing work, requires similar strategies to any practice change.

Paramedic students' experiences of stress whilst undertaking ambulance placements - An integrative review.

Work integrated learning (WIL) in the ambulance setting is an essential part of the paramedic curriculum. However, qualified ambulance personnel are reported to experience higher suicide rates and mental stress disorders due to high pressure work environments, and there is growing concern for the wellbeing of students entering this setting. The aim of this integrative review was to explore how studies have reported paramedic students' experience of stress whilst undertaking WIL. Five studies met the inclusion criteria and were evaluated for quality according to validated tools from the Critical Appraisal Skills Program, then ranked on the level of evidence used. Data was summarised in a comprehensive research paper matrix, and findings were categorised into levels and sources of stress. Levels of stress were measured by the percentage of paramedic students who developed post-traumatic stress disorder. The primary sources of stress were experiencing death and fear of making clinical mistakes. Students also identified emotional expression as a negative attribute. Future research should prioritise identifying the levels and sources of stress students face in each year of their academic program when undertaking WIL to provide a direction for preparatory activities that may mitigate the negative effects of stress.

Clinical trials with cannabis medicines-guidance for ethics committees, governance officers and researchers to streamline ethics applications and ensuring patient safety: considerations from the Australian experience.

With cannabis medicines now obtaining legal status in many international jurisdictions (generally on the authorisation of a medical professional), a rapid increase in consumer demand for access to cannabis as a therapeutic option in the treatment and management of a range of indications is being noted. Despite this accessibility, knowledge on optimal use is lacking. Further drug development and clinical trials at regulatory standards are necessary both if a better understanding of the efficacy of cannabis medicines, optimal product formulation and indication-specific dosing is needed and to ensure the broader quality and safety of cannabis medicines in the clinical setting.To enable this, clinical, academic and public calls for the undertaking of rigorous clinical trials to establish an evidence base for the therapeutic use of cannabis medicines have been made internationally. While this commitment to undertake human studies with cannabis medicines is welcomed, it has highlighted unique challenges, notably in the review stages of ethics and governance. This often results in lengthy delays to approval by Human Research Ethics Committees (herein 'HREC', Australia's nomenclature for Institutional Review Boards) and trial commencement. A principal concern in these cases is that in contrast to clinical trials using other more conventional pharmaceutical products, trials of cannabis medicines in humans often involve the use of an investigational product prior to some (or any) of the preclinical and pharmaceutical safety issues being established. This paucity of data around product safety, potential drug interactions, continuity of supply, shelf life and product storage results in apprehension by HRECs and governance bodies to endorse trials using cannabis medicines.This manuscript draws from the experiences of Australian researchers and staff involved in clinical trials of cannabis medicines to describe some of the common difficulties that may be faced in the HREC approval process. It also presents practical advice aimed to assist researchers, HRECs and governance officers navigate this complex terrain. While the authors' experiences are situated within the Australian setting, many of the barriers described are applicable within the international context and thus, the solutions that have been proposed are typically adaptive for use within other jurisdictions.

The impact of automated medicine dispensing units on nursing workflow: A cross-sectional study.

BACKGROUND: Automated medication dispensing cabinets are ubiquitous in hospitals in the United States and prevalent in Canada, but they are still relatively new to health services elsewhere. The automation of medication management using distributed dispensing units is aimed at improving stock management and patient safety; however, the evidence for the latter remains equivocal, and the impact on nursing workflow is poorly understood. OBJECTIVE: This study evaluated the impact on the nursing workflow of a distributed automated medication dispensing system. The research aimed to explore the acceptability and utility of this system in a variety of clinical settings and to investigate similarities and differences in the use of the dispensing cabinets across different clinical areas. DESIGN: A cross-sectional design was employed. SETTING: The setting was a newly constructed 450-bed regional Australian tertiary hospital. PARTICIPANTS: The study involved 174 registered nurses and 12 pharmacy assistant staff from general ward and specialty areas who were using the automated medication dispensing cabinets. METHODS: Methods included a hospital-wide survey of users and an observation study of nursing workflow around the automated medication cabinets in specific clinical areas. RESULTS: The majority of staff were satisfied with the system and were positive about the overall safety and security. Key concerns related to access delays, and increased time needed due to walking distance and interruptions from other staff. Staff perceived that the automated medication dispensing cabinet use slowed medication administration processes as a result of queueing, and it also had other impacts on workflow. The system was found to expedite processes around controlled/narcotic drug administration. Re-stocking requirements presented operational issues; pharmacy assistants were observed waiting for opportunities to complete re-stocking tasks in the face of competing clinical requirements. Nurses from general wards were more satisfied with the system than those from specialty areas. CONCLUSIONS: Automated medication dispensing cabinets were widely accepted by nurses in a large newly opened hospital in a variety of acute clinical areas despite disruptions to workflow. Adaptations for access were more acceptable to nurses in general wards than those in specialty areas prompting consideration of redesign to improve suitability. Tweetable abstract: Automated medication cabinets change nursing workflow because of queueing, interruptions from other staff and increased walking. Ward nurses are more accepting of such workflow disruptions than speciality area nurses #medicationsafety #nurseworkflow #nursesatisfaction (268 char).

The effects of a virtual learning environment compared with an individual handheld device on pharmacology knowledge acquisition, satisfaction and comfort ratings.

BACKGROUND: Virtual reality is reported to improve post-intervention knowledge and skills outcomes of health professionals compared to traditional teaching methods or digital online media. However, providing equitable access to high quality virtual reality resources for large, diverse nursing and midwifery student cohorts within multi-campus settings remains challenging. OBJECTIVES: This study compared the effect on student learning, satisfaction and comfort following exposure to a three-dimensional pharmacology artefact in a virtual facility (CAVE2™)1 with viewing of the same artefact using a mobile handheld device with stereoscopic lenses attached. DESIGN: The study used a pretest-posttest design. SETTING: School of Nursing and Midwifery in a regional university in Southeast Queensland, Australia. PARTICIPANTS: Two hundred and forty-nine second year undergraduate nursing and midwifery students. METHODS: Online multiple choice tests were deployed to measure knowledge acquisition. Self-reported satisfaction scores and comfort ratings were collected using questionnaires. RESULTS: Participants were not disadvantaged in terms of knowledge acquisition by using either CAVE2™ or the mobile handheld visualisation mode (P = 0.977). Significant differences in favour of the CAVE2™ environment were found in between students' satisfaction scores for clinical reasoning (P = 0.013) and clinical learning (P < 0.001) compared to the handheld mode, and there were no significant differences in their satisfaction with debriefing and reflective practice processes (P = 0.377) related to undertaking visualisation activities. A small number of students using handheld devices with stereoscopic lenses reported greater discomfort in relation to the visualisation that negatively impacted their learning (P = 0.001). CONCLUSION: Three-dimensional artefacts using mobile devices is promising in terms of cost-effectiveness and accessibility for students with restricted access to on-campus teaching modes.

'Speaking up for safety': A graded assertiveness intervention for first year nursing students in preparation for clinical placement: Thematic analysis.

BACKGROUND: The international literature highlights patient safety as a critical issue in contemporary health care. Poor interpersonal relationships and ineffective health care team communication are identified as dominant human factors contributing to clinical errors and adverse events. Of concern is that some students appear to lack the skills to speak up for themselves, their patients and others when witnessing unsafe practice on clinical placement. STUDY AIMS: To elicit student and staff perspectives on the quality, effectiveness and appropriateness of an assertiveness-based communication activity prior to clinical placement. DESIGN: The study used a qualitative design. A communication module was offered in the second week of the practice laboratory and involved the use of two graded assertiveness frameworks. SETTING: School of Nursing, Midwifery and Paramedicine, at a regional university in South East Queensland, Australia. PARTICIPANTS: Of the 535 first year undergraduate nursing students enrolled in the laboratory courses, 73 (13.6%) completed an anonymous online survey. Individual interviews were conducted with a university student wellbeing coordinator and student mentor to gain their perspectives of the data. METHODS: An evaluation survey with seven qualitative questions and individual semi-structured interviews were employed. Data was analysed using a thematic approach. RESULTS: Five major themes emerged from the survey data: vicarious learning, establishing boundaries, support in practice, advocacy awareness and practice utility. Three themes emerged from the interviews: empowerment and confidence; support and preparation for advocacy; and authentic learning. CONCLUSIONS: Findings indicate that teaching assertiveness skills and establishing a preparatory framework for 'speaking up for safety' early in a nursing students tertiary education can have important psychosocial implications for their confidence, empowerment and success. The activity provided an authentic learning experience with perceived practical application to the workplace and, has the potential to enhance first year curricula by improving communication tools used for students preparing for practice.

Effectiveness of three-dimensional visualisation on undergraduate nursing and midwifery students' knowledge and achievement in pharmacology: A mixed methods study.

BACKGROUND: Historically nursing and midwifery students have reported difficulty understanding the concept-based science underpinning the interactions between drugs and their targets. This knowledge is crucial for the administration and monitoring of the therapeutic and adverse effects of medications. Immersive three-dimensional technology is reported to enhance understanding of complex scientific concepts but the physical effects of motion sickness may limit its use. OBJECTIVES: This project compared the effectiveness of three-dimensional immersive visualisation technology with two-dimensional visualisation technology as a teaching method to improve student understanding of a pharmacological concept, and to assess levels of student discomfort and satisfaction associated with the experience. DESIGN: Traditional lecture content and presentation about drug-receptor binding was followed by exposure to either a two- or three-dimensional artifact visualising ß-adrenoceptor binding. Two student groups were compared by type of exposure: Group 1 watched the artifact via a three-dimensional immersive facility and Group 2 on a wide, two-dimensional screen. SETTINGS: School of Nursing and Midwifery in a regional university in Southeast Queensland, Australia. PARTICIPANTS: Two hundred and two second year undergraduate nursing and midwifery students. METHODS: The study used mixed methods methodology. Pre- and post- testing of student knowledge was collected using five multiple-choice questions. A post-intervention survey elicited students' self-assessed perceptions of discomfort and satisfaction with the learning experience. RESULTS: The three-dimensional immersive learning experience was comparable to the two-dimensional experience in terms of satisfaction and comfort but resulted in statistically significant improvements in post-test scores. CONCLUSIONS: The three-dimensional experience improved understanding when compared to two-dimensional viewing, satisfied students leaning needs, and caused minimal discomfort. The results are encouraging in terms of using three-dimensional technology to enhance student knowledge of pharmacological concepts necessary for competency in medication management.

Preparation for workplace adversity: Student narratives as a stimulus for learning.

Nursing students are not always well prepared for the kind of adverse events they may experience in the workplace and yet it seems apparent that future students could benefit from learning about such experiences so that they can be avoided, or their impact minimised. This research aimed to identify nursing students' experiences of adversity, collaborate with students to discern important lessons for future students in their experiences, and make recommendations for other educators on how to use these adversity stories as lessons. Seven Australian nursing students were interviewed using critical incident technique consisting of 7 questions. This paper focuses on the responses to the questions: "Does this story's message have a place in the curriculum?" and "How would you teach this lesson?" Data were analysed using critical discourse analysis. Four recurring discourses emerged including: power relationships are a two-way street; learn from mistakes to prevent mistakes; begin cultural consciousness-raising in first year, first semester; and become critically self-aware. Narratives derived from original stories of adversity may be a valuable source of learning about the realities of the workplace but to benefit fully, educators need to assist students to notice and analyse embedded messages.

Surveying the experiences and perceptions of undergraduate nursing students of a flipped classroom approach to increase understanding of drug science and its application to clinical practice.

Patient harm from medication error is a significant issue. Individual failures by health professionals including knowledge deficits and poor communication have been identified as increasing the likelihood of medication administration errors. In Australia, the National Strategy for Quality Use of Medicines in 2002 compels health professionals to have the knowledge and skills to use medicines safely and effectively. This paper examines nursing students' perceptions of the effectiveness of a flipped classroom approach to increase understanding of pharmacology principles and the application of this knowledge to medication practice. An internet-based self-completion questionnaire was used in 2013 (n = 26) after the flipped classroom approach was implemented, and pre- (n = 6) and post-flipping (n = 25) in 2014. Students who engaged with digitally recorded lectures (eLectures) prior to face-to-face workshops stated that they had greater understanding of the subject and enhanced critical thinking skills. The replay function of the eLecture was perceived by some students as most beneficial to independent learning. However, for some students, time constraints meant that they relied on eLectures alone, while others preferred traditional teaching methods. Although limited by sample size and potential participant bias, the results provide insights about the flipped classroom experience from a student perspective.

A Central Role for HLA-DR3 in Anti-Smith Antibody Responses and Glomerulonephritis in a Transgenic Mouse Model of Spontaneous Lupus.

MHC, especially HLA-DR3 and HLA-DR2, is one of the most important genetic susceptibility regions for systemic lupus erythematosus. Human studies to understand the role of specific HLA alleles in disease pathogenesis have been hampered by the presence of strong linkage disequilibrium in this region. To overcome this, we produced transgenic mice expressing HLA-DR3 (DRß1*0301) and devoid of endogenous class II (both I-A and I-E genes, AE(0)) on a lupus-prone NZM2328 background (NZM2328.DR3(+)AE(0)). Both NZM2328 and NZM2328.DR3(+)AE(0) mice developed anti-dsDNA and glomerulonephritis, but anti-dsDNA titers were higher in the latter. Although kidney histological scores were similar in NZM2328 and NZM2328.DR3(+)AE(0) mice (7.2 ± 4.3 and 8.6 ± 5.7, respectively, p = 0.48), the onset of severe proteinuria occurred earlier in NZM2328.DR3(+)AE(0) mice compared with NZM2328 mice (median, 5 and 9 mo respectively, p < 0.001). Periarterial lymphoid aggregates, classic wire loop lesions, and occasional crescents were seen only in kidneys from NZM2328.DR3(+)AE(0) mice. Interestingly, NZM2328.DR3(+)AE(0) mice, but not NZM2328 mice, spontaneously developed anti-Smith (Sm) Abs. The anti-Sm Abs were seen in NZM2328.DR3(+)AE(0) mice that were completely devoid of endogenous class II (AE(-/) (-)) but not in mice homozygous (AE(+/+)) or heterozygous (AE(+/-)) for endogenous MHC class II. It appears that only HLA-DR3 molecules can preferentially select SmD-reactive CD4(+) T cells for generation of the spontaneous anti-Sm immune response. Thus, our mouse model unravels a critical role for HLA-DR3 in generating an autoimmune response to SmD and lupus nephritis in the NZM2328 background.

Transformers: changing the face of nursing and midwifery in the media.

AIM: This paper reports an educational strategy designed to sensitise and empower students about the impact of media representations of nursing and midwifery on their public image. BACKGROUND: Numerous studies continue to reveal that stories about nursing and midwifery presented in the mainstream media are often superficial, stereotypical and demeaning. Inaccurate portrayals of nursing damage our professional reputation with the public and potential consumers. It also sends the wrong message to future nursing students. Images are a powerful conductor of misinformation, suggesting to others that nurses are not important agents for social change. METHODS: In 2012, a small team of academics designed a photography competition and judging process for undergraduate and postgraduate students of nursing and midwifery enrolled at a regional Australian university. RESULTS: The winning entries were photographs of high quality and conveyed rich meaning. They provide an interesting and positive counterpoint to derogatory images often propagated by mainstream media. CONCLUSION: There is benefit in extending this project so that it: appeals to more students, builds leadership skills, leads to wider social change and benefits society. The intention is to develop the process of student engagement as an educational intervention, and explore experiences and outcomes with stakeholders.

From me to we: transforming values and building professional community through narratives.

Entering high acuity environments for the first time is a daunting experience for nursing students and new graduates. They are challenged to be both highly skilled and critical thinkers. In addition, because of increasing acuity and the burgeoning prevalence of patients with long-term, multi-faceted health care issues, particularly in the elderly population, students also need advanced interpersonal communication skills. Nurse academics need to respond to these imperatives by examining the fundamental values of the profession so that they can provide learning opportunities that place equal emphasis on developing affective attributes alongside cognitive and psychomotor skills. This paper presents a novel values-based learning activity using transformative learning principles. Three extracts from a book were chosen that conveyed the uncertainty and insecurity that a novice Intensive care nurse overcame to become a competent, professional, trusted practitioner, her passion to be part of a caring profession and the positive role models who shaped her values. Transformative Learning questions were developed to promote critical reflection on the shared values of the profession and the transition from the personal to professional self. Students' insights from the activity focused on their aspirations to provide patient-centred care and included recognition of the emotional labour of caring, the need to rise above negative cultures, how to challenge out-dated practices and the importance of strengthening professional identity. Krathwohl et al. (1964) Stages of Affective Learning was used to evaluate the activity.

Advancing affective attributes and empowering undergraduate students--lessons learned from the Bali bombing.

Caring as an integral component in the nursing curriculum is enjoying a resurgence in the literature of late. The argument is that nursing education has tended to overemphasise the cognitive domain and under emphasise the affective. An alternative is to use the combined effect of cognition, imagination, intuition and emotion. This is supported by the theory of transformational learning, whereby students clarify their personal and professional purpose in life and are empowered to become informed, self-efficacious practitioners and autonomous thinkers as they negotiate personal values and meaning. In order to integrate these important theoretical concepts into everyday practice, educators need practical examples and case studies that show how caring is taught. This paper continues the conversation on narrative and transformational learning pedagogies and illustrates how affective attributes are developed through a single lecture. The aim of the lecture was to sensitise students to the human impact of terrorism and violence and the effects on both health care workers and the survivors of trauma. The rationale was that by allowing students to critically reflect on their own core knowledge and skills, they could question prior perceptions of their role, resulting in a revised or new perspective of those experiences and strengthen their belief in their abilities to cope in crisis situations. This transformative approach involved the delivery of knowledge and theory underpinning disaster response, personal narratives about a critical learning event that embodied clinically relevant lessons, activities that promoted critical self-reflection to strengthen students' beliefs in their own ability to cope by converting core knowledge into action and, finally student evaluation of the lesson (see Table 1).

Differential responses to Smith D autoantigen by mice with HLA-DR and HLA-DQ transgenes: dominant responses by HLA-DR3 transgenic mice with diversification of autoantibodies to small nuclear ribonucleoprotein, double-stranded DNA, and nuclear antigens.

Anti-Smith (Sm) D autoantibodies are specific for systemic lupus erythematosus. In this investigation, the influence of HLA-D genes on immune responses to SmD was investigated. Mice with HLA-DR3, HLA-DR4, HLA-DQ0601, HLA-DQ0604, or HLA-DQ8 transgenes were immunized with recombinant SmD1, and their Ab responses were analyzed. Analysis by ELISA showed that all strains responded well to SmD. However, when synthetic SmD peptides were used as substrate, DR3 mice had the highest Ab response followed by DQ8, DQ0604, DQ0601, and DR4. A similar trend was observed in Western blot analysis using WEHI 7.1 cell lysate as the substrate, with the exception that DR4 mice did not generate detectable amounts of Abs. Only sera from DR3 and DQ0604 mice immunoprecipitated A-ribonucleoprotein (RNP), SmB, and SmD. Intermolecular epitope spreading to A-RNP and SmB was evident in DR3 and DQ0604 mice, as sera depleted of anti-SmD Abs were reactive with these proteins. DR3 mice also generated an immune response to C-RNP. Anti-nuclear Abs were detected in the majority of the DR3 mice, whereas moderate reactivities were seen in DQ0604 and DQ8 mice. Interestingly, only DR3 mice mounted an anti-dsDNA Ab response. Approximately half of the anti-dsDNA Abs were cross-reactive with SmD. Ab responses correlated with the strength of the T cell responses. Thus, HLA-DR3 appears to be the dominant HLA-D gene that determines the magnitude and quality of the anti-SmD immune response. In addition, our findings provide insights into the origin of the anti-dsDNA Abs often detected in patients with systemic lupus erythematosus.

Hormone treatment enhances WT1 activation of Renilla luciferase constructs in LNCaP cells.

The zinc finger transcription factor, WT1, regulates many growth control genes, repressing or activating transcription depending on the gene and cell type. Based on earlier analyses of the effect of WT1 on androgen responsive genes, we hypothesized that there may be an interaction between the androgen signaling pathway and WT1, such that the commonly used Renilla luciferase control vectors were activated in LNCaP prostate cancer cells. Using cotransfection assays we tested the effects of WT1 and/or the androgen analog, R1881, on two Renilla luciferase vectors, pRL-SV40 and the promoter-less pRL-null. To determine whether the zinc finger DNA binding domain was required, the zinc finger mutant DDS-WT1 (R394W) was tested; but it had no significant effect on the Renilla luciferase vectors. To determine whether the androgen signaling pathway was required, WT1 was co-transfected with Renilla vectors in cells with varied hormone responsiveness. The WT1 effect on pRL-null varied from no significant effect in 293 and PC3 cells to very strong enhancement in LNCaP cells treated with 5 nM R1881. Overall, these results suggest that hormone enhanced WT1 mediated activation of Renilla luciferase and that these interactions require an intact WT1 zinc finger DNA binding domain.

Regulation of vascular endothelial growth factor, VEGF, gene promoter by the tumor suppressor, WT1.

Understanding angiogenesis and growth control is central for elucidating prostate tumorigenesis. However, the mechanisms of activation of the angiogenic gene, vascular endothelial growth factor (VEGF) are complex and its regulation in prostate cancer is not well understood. In previous studies, VEGF expression levels were correlated with altered levels of the zinc finger transcription factor, WT1. Since the VEGF promoter has several potential WT1 binding sites and WT1 regulates many growth control genes, here we assessed whether WT1 might also regulate VEGF transcription. Using transfection and DNA binding assays, functional WT1 binding sites were localized within the proximal VEGF promoter. Transfection of the DDS-WT1 (R394W) zinc finger mutant had no significant effect on VEGF-luciferase reporter activity, suggesting that an intact zinc finger DNA binding domain was required. Interestingly, WT1-mediated regulation of VEGF reporter constructs varied in different cell types. In androgen-responsive, LNCaP prostate cancer cells, hormone treatment enhanced WT1-mediated activation of the VEGF promoter constructs. Overall, these results suggest that WT1 transcriptionally regulates VEGF through interaction of its zinc finger DNA binding domain with the proximal GC-rich VEGF promoter. These findings may shed light on the role of WT1 in angiogenesis and prostate cancer progression.