Cellular uptake of ribonuclease A-functionalised core-shell silica microspheres.

Analysis of protein function in a cellular context ideally requires physiologically representative levels of that protein. Thus conventional nucleic acid-based transfection methods are far from ideal owing to the over expression that generally results. Likewise, fusions with protein transduction domains can be problematic whilst delivery via liposomes/nanoparticles typically results in endosomal localisation. Recently, polymer microspheres have been reported to be highly effective at delivering proteins into cells and thus provide a viable new alternative for protein delivery (protein transduction). Herein we describe the successful delivery of active ribonuclease A into HeLa cells via novel polymer core-silica shell microspheres. Specifically, poly(styrene-co-vinylbenzylisothiouronium chloride) core particles, generated by dispersion polymerisation, were coated with a poly(styrene-co-trimethoxysilylpropyl methacrylate) shell. The resultant core-shell morphology was characterised by transmission electron, scanning electron and confocal fluorescence microscopies, whilst size and surface charge was assessed by dynamic light scattering and zeta-potential measurements, respectively. Subsequently, ribonuclease A was coupled to the microspheres using simple carbodiimide chemistry. Gel electrophoresis confirmed and quantified the activity of the immobilised enzyme against purified HeLa RNA. Finally, the polymer-protein particles were evaluated as protein-transduction vectors in vitro to deliver active ribonuclease A to HeLa cells. Cellular uptake of the microspheres was successful and resulted in reduced levels of both intracellular RNA and cell viability.

Multistrain influenza protection induced by a nanoparticulate mucosal immunotherapeutic.

All commercial influenza vaccines elicit antibody responses that protect against seasonal infection, but this approach is limited by the need for annual vaccine reformulation that precludes efficient responses against epidemic and pandemic disease. In this study we describe a novel vaccination approach in which a nanoparticulate, liposome-based agent containing short, highly conserved influenza-derived peptides is delivered to the respiratory tract to elicit potent innate and selective T cell-based adaptive immune responses. Prepared without virus-specific peptides, mucosal immunostimulatory therapeutic (MIT) provided robust, but short-lived, protection against multiple, highly lethal strains of influenza in mice of diverse genetic backgrounds. MIT prepared with three highly conserved epitopes that elicited virus-specific memory T-cell responses but not neutralizing antibodies, termed MITpep, provided equivalent, but more durable, protection relative to MIT. Alveolar macrophages were more important than dendritic cells in determining the protective efficacy of MIT, which induced both canonical and non-canonical antiviral immune pathways. Through activation of airway mucosal innate and highly specific T-cell responses, MIT and MITpep represent novel approaches to antiviral protection that offer the possibility of universal protection against epidemic and pandemic influenza.

Effect of X- and Y-box deletions on the development of diabetes in H-2Ealpha-chain transgenic nonobese diabetic mice.

The development of type 1 diabetes in nonobese diabetic (NOD) mice is influenced by major histocompatibility complex (MHC) class II genes. The NOD-E transgenic mouse, which expresses H2-E as a result of the introduction of an Ead gene, is protected from development of type 1 diabetes. While the mechanism of protection remains unclear, the effect has been regarded as a model system for MHC protection from autoimmunity. We have investigated the effect of deletions of the Ea promoter region, which, in turn, affect H2-E expression patterns in transgenic NOD mice. We have constructed transgenic NOD mice where the X (DeltaX) and Y (DeltaY) boxes of the Ead gene have, respectively, been functionally deleted. Previous reports, using X- or Y-box-deleted H2-E transgenic mice, made by crossing the appropriate transgenes onto the NOD background from C57BL/6 transgenic mice, indicated that promoter mutation abrogated the H2-E-mediated protection seen in NOD-E. The NOD DeltaX and NOD DeltaY transgenic mice generated in the present study differ in susceptibility to diabetes from wild-type NOD mice. NOD DeltaY1 animals are protected from diabetes development, while DeltaX mice remain susceptible, albeit to a lesser extent than the parental NOD strain.

Stephen Abrahamson, PhD, ScD, educationist: a stranger in a kind of paradise.

This profile of Stephen Abrahamson, Ph.D., Sc.D., is the first of six profiles to appear as part of the Exemplar project focused of six retired medical educators who transformed the field of medical education. The exemplars, all graduate degree recipients in education were interviewed by six senior present-day medical educators using a common protocol designed to elicit career chronology and the significant contributions of educationists to medical education of Dr. Abrahamson's profile was based on an in-depth two-day interview, examination of a comprehensive list of his publications, the history of the Society of the Directors of Research in Medical Education, and unsolicited conversations with several of his colleagues. Dr. Abrahamson began his career teaching high school, later receiving a masters and doctorate degrees, in preparation for a career as a teacher-educator. Through collaboration with Dr. George Miller, Dr. Abrahamson began his career as an educationist in medicine - one who studies the education process and prepares others to become teachers - by teaching medical school faculty about the science of education. Dr. Abrahamson's career was devoted to applying his evidence-based education approach to the newly emerging profession of medical education. An examination of his career shows that he made four vital contributions to medical education - defining the educationist role, serving as a teaching/mentor/network builder/friend to medical educators, curriculum change agent and innovator at USC, and demonstrating and articulating the value of offices of medical education and research in medical education. More broadly, Dr. Abrahamson identified three major contributions made by educationists to the field of medical education: the application of education principles to instructional/assessment innovations (e.g., programmed patients), an evidence-based approach to assessing education, and faculty development/teacher training. Based on his half-century of experience in medical education, Dr. Abrahamson outlined seven lessons for success as an educationist in medicine.

Lessons learned from a distance-based consulting program to assist faculty development projects.

Changes in faculty roles and demographics necessitate a re-examination of the types of professional development opportunities offered in academic institutions. A distance-based consulting program was designed to assist faculty development projects as they progress through all stages of faculty development: needs assessment, project design, implementation, and, in particular, program evaluation and dissemination of results (i.e., presentations and published articles). The progress of 17 faculty development projects in primary care educational sites that received assistance in the United States and Canada was tracked over two years. Three factors were identified as having the most impact on the success of faculty development projects: (1) funds committed to and designated for faculty development; (2) funded, protected time for at least one person to implement the faculty development initiative; and (3) an environment capable of supporting faculty development initiatives (e.g., no major budget shortfall, few faculty transitions, a strong mission, no threat of mergers). Only a few of the participating sites reached the stage of evaluating and publishing articles about the outcomes of their projects within the designated 15-month time frame, with many sites reporting environmental impediments to project success. The authors describe the institutional characteristics that facilitated project success, assess the usefulness of distance-based consulting efforts, and offer recommendations for future distance-based consulting programs. They conclude by noting that the personal touch (i.e., one-on-one contact with consultants) is what is most appreciated, and that excellent one-on-one, in-person assistance may be inherently more effective than even the best-run distance-based consulting.

Partial uptake of varicella vaccine and the epidemiological effect on varicella disease in 11 day-care centers in North Carolina.

BACKGROUND: The increasing use of varicella vaccine in children attending day care has rapidly decreased the incidence of wild-type varicella disease. The herd immunity noted is significant and will have an effect on the epidemiology of natural varicella. OBJECTIVE: To monitor the change in varicella incidence in day-care attendees after the licensure of varicella vaccine. DESIGN: A prospective observational cohort study design. SETTING: Eleven private day-care centers and preschools in North Carolina participated in the study from January 1, 1995, through December 31, 1999. PARTICIPANTS: All children in the 11 centers were eligible for participation. Some participated more actively, supplying information on a regular basis. Others participated passively. Day-care personnel provided information about all cases of varicella. INTERVENTIONS: None. MAIN OUTCOME VARIABLES: The change in the incidence of varicella disease was documented as the use of varicella vaccine increased. RESULTS: Varicella vaccine coverage increased substantially from 4.4% in 1995 to 63.1% in December 1999. The vaccination rate accelerated dramatically in 1996 and 1997, leveled off in 1998, and rose again in 1999. Cumulative varicella incidence decreased from 16.74 cases per 1000 person-months in July 1996 to 1.53 cases per 1000 person-months in December 1999 in unvaccinated children. CONCLUSIONS: The varicella vaccination rate continued to increase slowly in the day-care population after an initial rapid uptake. The decrease in varicella disease is greater than the increase in varicella vaccination. This herd effect is welcome and even apparent in the unvaccinated children younger than 1 year.

Curricular change in medical schools: how to succeed.

Society's changing needs, advancing knowledge, and innovations in education require constant changes of medical school curricula. But successful curricular change occurs only through the dedicated efforts of effective change agents. This study systematically searched and synthesized the literature on educational curricular change (at all levels of instruction), as well as organizational change, to provide guidance for those who direct curricular change initiatives in medical schools. The focus was on the process of planning, implementing, and institutionalizing curricular change efforts; thus, only those articles that dealt with examining the change process and articulating the factors that promote or inhibit change efforts were included. In spite of the highly diverse literature reviewed, a consistent set of characteristics emerged as being associated with successful curricular change. The frequent reappearance of the same characteristics in the varied fields and settings suggests they are robust contributors to successful change. Specifically, the characteristics are in the areas of the organization's mission and goals, history of change in the organization, politics (internal networking, resource allocation, relationship with the external environment), organizational structure, need for change, scope and complexity of the innovation, cooperative climate, participation by the organization's members, communication, human resource development (training, incorporating new members, reward structure), evaluation, performance dip (i.e., the temporary decrease in an organization's performance as a new program is implemented), and leadership. These characteristics are discussed in detail and related specifically to curricular change in medical school settings.

"No fear" curricular change: monitoring curricular change in the W. K. Kellogg Foundation's National Initiative on Community Partnerships and Health Professions Education.

PURPOSE: To assess the effect of the W. K. Kellogg Foundation's five-year (9/91 to 9/96) primary care curricular change initiative involving seven sites and 27 schools with respect to courses offered, graduates' specialty choices, and valued school features (e.g., reputation, research, productivity, school climate) and to track how well the schools possessed or developed the processes associated with enduring curricular change. METHOD: Information was collected via pre- and post-surveys of faculty and students, site visits, annual reports, and medical students' specialty match results. RESULTS: In general, the schools either possessed or developed the institutional, curricular design, and curriculum features associated with successful curricular change. Further, the initiative had a positive or neutral impact on most of the valued features: 199 courses were revised or developed, including 138 offered in the community, involving 141 interdisciplinary faculty. Comparing 1991-1997 data, the average percentage increases in students' selecting primary care from the involved schools were greater than the national percentages. In 1997, a larger percentage of project school graduates selected primary care than the national average. CONCLUSION: Medical schools can make major curricular changes and achieve intended outcomes (e.g., new and/ or revised courses, multidisciplinary instruction, instruction in the community, and changes in specialty choice), and this can occur without negatively impacting other valued school aspects.

LPN role advancement: from blueprints to ribbon cutting.

The role of the Licensed Practical Nurse (LPN) in an acute care setting has been a point of debate for many years. Increasing LPN job responsibility has not been widely reported. In this article, the authors describe the process used to advance the LPN role. Emphasized here is the educators' contribution to the process from inception to implementation.

Postlicensure study of varicella vaccine effectiveness in a day-care setting.

BACKGROUND: Varicella vaccine has been licensed for use in the United States since the spring of 1995. The acceptance of the vaccine and its effect on varicella incidence in children is important. AIM: To document the effectiveness of the varicella vaccine in children attending day care in 11 centers in North Carolina. METHODS: A dynamic cohort study design was used in 11 day-care centers in North Carolina. Multiple cross-sectional evaluations were performed and children were noted to be vaccinated or not and diseased or not. Vaccine effectiveness was estimated by comparing the varicella attack rate in the vaccinated with the varicella attack rate in the unvaccinated. Person time was used as the denominator for all calculations. RESULTS: During the study period February 1, 1996, to September 1, 1997, 134 cases of varicella occurred in the unvaccinated and 11 cases occurred in the vaccinated children. The attack rates in the vaccinated and unvaccinated were 2.49 and 14.66, respectively, for an overall vaccine effectiveness of 83% for mild/moderate disease. CONCLUSIONS: In the day-care setting varicella vaccine demonstrated benefit in preventing and modifying wild-type varicella disease.

Leadership behaviors for successful university--community collaborations to change curricula.

PURPOSE: What constitutes effective leadership in a collaborative effort to achieve enduring curricular and student career changes? This question was investigated as part of a larger evaluation of the W. K. Kellogg Foundation's Community Partnership Health Professions Education, a five-year initiative involving projects at seven sites. The goal was to produce more primary care health providers by making enduring curricular change. METHOD: Data were collected from participants with respect to predictors of project success and leaders' use of 16 behaviors via telephone interviews, mailed surveys, and focus groups. Focus groups also gathered project leaders' views of skills and knowledge necessary for effective leadership. RESULTS: Leadership strategies associated with positive outcomes were: consistent leader; use of multiple cognitive frames, especially a human resource frame; use of a broad range of leadership behaviors, particularly participative governance and cultural influence; and a majority of community representatives on the partnership board. The primary leader, compared with a leadership team, is most influential in achieving positive outcomes. CONCLUSION: Effective leaders use a broad array of behaviors, but particularly emphasize the use of participative governance and culture/value-influencing behaviors. In addition, the more frequent use of these behaviors compared with the use of organizational power behaviors is important. It is helpful to perceive the project from a human-relations frame and at least one other frame. Using a leadership team can be helpful, especially in building coalitions, but the importance of the primary leader's behaviors to project outcomes is striking.

When (not if) evaluation flexibility is desirable. Examples from the CPHPE initiative.

The evaluation literature often debates whether evaluators should be flexible in evaluation design and activities in order to collaborate with program directors and be responsive to programming needs. Two conditions are specified under which evaluation flexibility is not only desirable but essential. Two examples from the cluster evaluation of the W. K. Kellogg Foundation's Community Partnerships for Health Professions Education initiative are provided to illustrate why flexibility under these conditions proved to be essential. One of the examples, related to the "community" involvement in the initiative, illustrates the need for flexibility as programs experience goals clarification. The other example, related to the coincidental national health care reform efforts, illustrates the need for flexibility both to capture programs' efforts to protect their integrity and to ensure against spurious conclusions as a result of external turbulence in policy environments. How the cluster evaluation team addressed these issues is also described.

Concurrent administration of inactivated hepatitis A vaccine with immune globulin in healthy adults.

301 healthy adult volunteers were randomized to one of three treatment groups: inactivated hepatitis A vaccine alone; inactivated hepatitis A vaccine with immune globulin (Ig) concurrently; or Ig alone. The first two treatment groups received a second dose of hepatitis A vaccine at week 24. Anti-HAV was measured 4, 8, 12, 24 and 28 weeks after the primary immunization. When comparing subjects receiving inactivated hepatitis A vaccine alone to those receiving vaccine and Ig, the seropositivity rates were not significantly different at 4, 8, 12 and 28 weeks, but at week 24 the seropositivity rate was lower in the group receiving both vaccine and Ig compared to the group receiving vaccine alone (92.0% compared to 97.0%). At weeks 8, 12 and 24 the geometric mean titers (GMTs) were significantly lower for subjects receiving both vaccine and Ig. The GMTs were not significantly different after the second dose of vaccine. At all time points, the lower serum antibody concentrations observed in subjects receiving both inactivated hepatitis A vaccine and Ig were nevertheless substantially higher than the cutoff for assay seropositivity and much higher than after Ig alone; these differences are therefore clinically insignificant.

Multidisciplinary education outcomes of the W.K. Kellogg Community Partnerships and Health Professions Education initiative.

This evaluation highlights several points to be considered by others instituting multidisciplinary approaches to health professions education. Community-based, multidisciplinary experiences appear to reinforce support students' interests in pursuing primary care careers. The multidisciplinary, community-based approach to health professions education did not affect academic learning. Project leaders and students reported no risks in terms of board scores between CPHPE students and others in traditional programs. The multidisciplinary, community-based approach to health professions education created opportunities at some sites for students to see "team medicine" in action. It was practical and helped students to understand how they could be more effective in their roles as opposed to competitive. Students require socialization within their own individual disciplines as well as socialization across disciplines. The differences in the structures of traditional health professions education schools interfered with the development of multidisciplinary contexts for learning at some sites. Campus faculty were not necessarily socialized to engage in multidisciplinary efforts. Their disciplines generally do not recognize and reward this behavior. Early and continuous faculty development may significantly help projects to improve communication and develop a better understanding of the contexts of curricular changes across disciplines. This evaluation was exploratory. Further research is needed to better understand what forms of multidisciplinary curriculum are most effective and economically feasible, what forms survive over time, and whether the intended final outcomes of the CPHPE initiative are achieved, not only with medical students but also with the other health professions students.

Sensitizing T-lymphocytes for adoptive immunotherapy by vaccination with wild-type or cytokine gene-transduced melanoma.

BACKGROUND: For the relatively nonimmunogenic B16-F10 murine melanoma, it has been found that genetically engineered expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) but not interleukin (IL)-2, IL-4, or interferon-gamma (IFN-gamma) resulted in a vaccine that could induce resistance to rechallenge. Because T cells from lymph nodes draining the sites of some progressive tumors can mediate tumor regression after in vitro activation, it seemed possible that even apparently nonimmunogenic melanoma cells might induce similar preeffector cells in the vaccine-draining lymph nodes (DLNs). METHODS: C57BL/6 mice were vaccinated with B16-F10 cells that were either unmodified or genetically modified to produce IL-2, IL-4, GM-CSF, or IFN-gamma. DLNs were harvested 10 days after vaccination for adoptive immunotherapy (AIT). The DLN cells were activated with bryostatin 1 and ionomycin (B/I), expanded for 10 days in culture, and transferred to mice with 3-day pulmonary metastases. Pulmonary nodules were counted 14 days after AIT. RESULTS: Adoptive transfer of expanded DLN lymphocytes sensitized by inoculation of WT B16-F10, or IL-4, GM-CSF, or IFN-gamma expressing cells significantly reduced pulmonary metastases. Despite the spontaneous regression of IL-2-transduced B16-F10 tumors, DLN from mice inoculated with IL-2 producing B16 cells had little or no antitumor activity. CONCLUSIONS: B16-F10 vaccination strategies that apparently do not induce systemic immunity can effectively sensitize DLN preeffector cells.

Beyond corporate-style downsizing: a better way for medical schools to succeed in a changing world.

There is a critical need for medical schools and universities to consider strategies beyond corporate-style downsizing to address revenue needs and reposition their organizations. The author presents considerable evidence and three reasons to reject downsizing as a way to facilitate long-term organizational success. Instead, she recommends that institutions use a comprehensive approach to individual and organizational development to assure a flexible, enduring organization. Specifically, medical schools and universities should take an institution-wide perspective and approach to continually training, retraining, or reassigning faculty and should continually adapt their organizational structures and procedures as necessary to achieve changing institutional goals. The result will be the retention of able and dedicated faculty, who will be crucial in helping their schools continue to be successful while adapting to a changing world.

Future faculty development in family medicine.

BACKGROUND AND OBJECTIVES: Several recurring conclusions about the changed conditions and future needs of faculty emerged from the articles in this dedicated issue. Faculty work is becoming more differentiated; time is increasingly constrained; and faculty are working in more diverse settings. Faculty reward structures and career ladders are changing, and acquiring additional competencies in new areas is essential. Although less then anticipated, there is some research to guide the development of programs to prepare faculty for these changes. Based on these conclusions, it is expected that in the future, faculty and program chairs will seek a tight match between what a faculty development effort addresses and the responsibilities of faculty participants and/or the goals of the department or program. The following trends are expected: 1) Regional/national comprehensive programs will often be the best choice for new faculty since many departments and residencies will not have sufficient resources to support their own faculty development programs. 2) Research-oriented faculty will be expected to devote 2 years or more to formal training. 3) Experienced faculty and department/program leaders, many of whom are graduates of our faculty development programs, will need advanced training in targeted areas, through on-site, contextualized education or through individualized programs--perhaps electronic. 4) Organizational development will be needed to help departments and programs make changes in missions and processes and to maintain a sense of community. At the same time that new models must be created to meet these future faculty development needs, support for such programs has decreased. Both local and national sources must be recommitted to support faculty development programs and research so we may advance our understanding of cost-effective approaches to faculty development. Funding agencies need to change their guidelines to match the current needs for development and limit funding to programs designed on evidence.