Acquisition of Medical Immunology Knowledge: A Preliminary Study of the Knowledge Structures of Medical Students

Medical students from both Duke-NUS and NUS participated in a study that attempted to assess their knowledge structure in the medical immunology domain. Students had to perform a sorting task with a list of concepts derived from immunology experts. We collected demographic information as well as sorting data and the diversity of the sorts are presented in this article

The future is in the numbers: the power of predictive analysis in the biomedical educational environment

Biomedical programs have a potential treasure trove of data they can mine to assist admissions committees in identification of students who are likely to do well and help educational committees in the identification of students who are likely to do poorly on standardized national exams and who may need remediation. In this article, we provide a step-by-step approach that schools can utilize to generate data that are useful when predicting the future performance of current students in any given program. We discuss the use of linear regression analysis as the means of generating that data and highlight some of the limitations. Finally, we lament on how the combination of these institution-specific data sets are not being fully utilized at the national level where these data could greatly assist programs at large

Association of Epstein-Barr virus with nasopharyngeal carcinoma and current status of development of cancer-derived cell lines

It is well known that the Epstein-Barr virus (EBV) contributes directly to tumourigenesis in nasopharyngeal carcinoma (NPC), primarily in the undifferentiated form of NPC (WHO type III; UNPC or UC), which is commonly found in South East Asia. Unfortunately, research in NPC has been severely hampered by the lack of authentic EBV-positive (EBV+) human NPC cell lines for study. Since 1975, there have been more than 20 reported NPC cell lines. However, many of these NPC-derived cell lines do not express EBV transcripts in long-term culture, and therefore that finding may dispute the fundamental theory of NPC carcinogenesis. In fact, currently only one EBV+ human NPC cell line (C-666) in long-term culture has been reported. Hence, most of the NPC cell lines may not be representative of the disease itself. In order to better understand and treat NPC, there is an urgent need to develop more EBV+ human NPC cell lines. In this review, we discuss the authenticity of existing NPC cell lines and the impact of our understanding of NPC biology on the treatment of the disease and the relationship of EBV to NPC in the context of cell lines

Ku86 exists as both a full-length and a protease-sensitive natural variant in multiple myeloma cells

Background: Truncated variants of Ku86 protein have previously been detected in 86% to 100% of freshly isolated patient multiple myeloma (MM) cells. Since, the Ku70/Ku86 heterodimer functions as the regulatory subunit of the DNA repair enzyme, DNA-dependent protein kinase, we have been interested in the altered expression and function of Ku86 variant (Ku86v) proteins in genome maintenance of MM. Results: Although, a number of studies have suggested that truncated forms of Ku proteins could be artificially generated by proteolytic degradation in vitro in human lymphocytes, we now show using whole cell immunoblotting that the RPMI-8226 and SGH-MM5 human MM cell lines consistently express full-length Ku86 as well as a 69-kDa Ku86v; a C-terminus truncated 69-kDa variant Ku86 protein. In contrast, Ku86v proteins were not detected in the freshly isolated lymphocytes as was previously reported. Data also indicates that the Ku86v was not generated as a result of carbohydrate modification but that serine proteases may act on the full-length form of the protein. Conclusion: These data confirm that MM cells contain bona fide Ku86v proteins that were generated intracellularly by a post-transcriptional mechanism, which required proteolytic processing

A Disease-Based Approach to the Vertical and Horizontal Integration of a Medical Curriculum

As medical disciplines have become increasingly interdisciplinary and evidenced-based medicine is widely practiced, there is a need for curricula that reflect these changes. The newly revised LCME standards 1.1 Strategic Planning and Continuous Quality Improvement and 8.3 Curricular Design, Review, Revision/Content Monitoring require ongoing curricular review to assure accreditation compliancy. We have completed a comprehensive review of our curriculum and have moved from a discipline-based curriculum to that of one that focuses on a systems/disease-based model. The approach allows for a more horizontally integrated curriculum in the preclinical years, while the use of 115 distinct disease and eight themes creates a quality assurance mechanism that allows for tracking of vertical integration across the entire curriculum. The first step in the development of this quality assurance model was to establish and empower a newly formed integration subcommittee. This subcommittee was tasked with developing a model to review, track and improve the horizontal and vertical integration of the curriculum. Our integrated curriculum is now in its second year having completed the initial identification of gaps and redundancies through a process that relies on the mapping of diseases and themes throughout the courses. This ongoing review and evaluation process has created a dynamic quality assurance process that allows our faculty to address issues of both horizontal and vertical integration of our curriculum at the course level

Predicting Medical Student Success on Licensure Exams

Many schools seek to predict performance on national exams required for medical school graduation using prematriculation and medical school performance data. The need for targeted intervention strategies for at-risk students has led much of this interest. Assumptions that preadmission data and high stakes in-house medical exams correlate strongly with national standardized exam performance needs to be examined. Looking at prematriculation data for predicting USMLE Step 1 performance, we found that MCAT exam totals and math-science GPA had the best prediction from a set of prematriculation values (adjusted R 2 = 11.7 %) for step 1. The addition of scores from the first medical school exam improved our predictive capabilities with a linear model to 27.9 %. As we added data to the model, we increased our predictive values as expected. However, it was not until we added data from year 2 exams that we started to get step 1 prediction values that exceeded 50 %. Stepwise addition of more exams in year two resulted in much higher predictive values but also led to the exclusion of many early variables. Therefore, our best step 1 predictive value of around 76.7 % consisted of three variables from a total of 37. These data suggest that the preadmission information is a relatively poor predictor of licensure exam performance and that including class exam scores allows for much more accurate determination of students who ultimately proved to be at risk for performance on their licensure exams. The continuous use of this data, as it becomes available, for assisting at-risk students is discussed (251)

Use of Phage Display to Isolate Specifi c Human Monoclonal Antibody Fragments Against a Potential Target for Multiple Myeloma

Introduction: Multiple myeloma (MM), a malignancy of plasma cells, accounts for 10% of all haematological malignancies and is currently incurable. Although it can be treated, the disease tends to relapse after several years and becomes increasingly resistant to conventional therapy. Investigations into using humoral therapy for MM are now underway with a view that novel therapeutic agents may provide a more targeted therapy for MM. Materials and Methods: Here, phage display, a faster and more efficient method compared to classical hybridoma fusion technology, was used as a proof-of-concept to isolate several single-chain Fragment variables (scFv) against Ku86. Results: Anti-Ku86 polyclonal scFvs biopanning was successful where third round scFvs (A450~1.1) showed a 1/3 increase in binding as compared to the first round scFvs (A450~0.4) with 100ug/mL of antigen (purified human Ku86). Subsequent selection and verifi cation of monoclonal antibodies using third round biopanning revealed 4 good affi nity binding clones ranging from A450~0.1 to A450~0.15 on 12.5ug/mL of antigen as compared to low binders (A450~0.07) and these antibodies bind to Ku86 in a specifi c and dose-dependent manner. Comparative studies were also performed with commercially available murine antibodies and results suggest that 2 of the clones may bind close to the following epitopes aa506-541 and aa1- 374. Conclusions: These studies using phage display provide an alternative and viable method to screen for antibodies quickly and results show that good affinity antibodies against Ku86 have been successfully isolated and they can be used for further studies on MM and form the basis for further development as anti-cancer therapeutic agents

Use of ultraviolet-light irradiated multiple myeloma cells as immunogens to generate tumor-specific cytolytic T lymphocytes

10.1186/1476-8518-6-2Journal of Immune Based Therapies and Vaccines6

Use of Ultraviolet Light Irradiated Multiple Myeloma Cells as Immunogens to generate Tumor Specific Cytolytic T Lymphocytes

Background: As the eradication of tumor cells in vivo is most efficiently performed by cytolytic Tlymphocytes (CTL), various methods for priming tumor-reactive lymphocytes have been developed. In this study, a method of priming CTLs with ultraviolet (UV)-irradiated tumor cells, which results in termination of tumor cell proliferation, apoptosis, as well as upregulation of heat shock proteins (HSP) expression is described. Methods: Peripheral blood mononuclear cells (PBMC) were primed weekly with UV-irradiated or mitomycin-treated RPMI 8226 multiple myeloma cells. Following three rounds of stimulation over 21 days, the lymphocytes from the mixed culture conditions were analyzed for anti-MM cell reactivity. Results: By day 10 of cultures, PBMCs primed using UV-irradiated tumor cells demonstrated a higher percentage of activated CD8+/CD4- T lymphocytes than non-primed PBMCs or PBMCs primed using mitomycin-treated MM cells. Cytotoxicity assays revealed that primed PBMCs were markedly more effective (p \u3c 0.01) than non-primed PBMCs in killing RPMI 8226 MM cells. Surface expression of glucose regulated protein 94 (Grp94/Gp96) and Grp78 were both found to be induced in UV-treated MM cells. Conclusion: Since, HSP-associated peptides are known to mediate tumor rejection; these data suggest that immune-mediated eradication of MM cells could be elicited via a UV-induced HSP process. The finding that the addition of 17-allylamide-17-demethoxygeldanamycin (17AAG, an inhibitor of HSP 90-peptide interactions) resulted in decreased CTL-induced cytotoxicity supported this hypothesis. Our study, therefore, provides the framework for the development of anti-tumor CTL cellular vaccines for treating MM using UV-irradiated tumor cells as immunogens

Ca2+ signaling modulates cytolytic T lymphocyte effector functions

Cytolytic T cells use two mechanisms to kill virally infected cells, tumor cells, or other potentially autoreactive T cells in short-term in vitro assays. The perforin/granule exocytosis mechanism uses preformed cytolytic granules that are delivered to the target cell to induce apoptosis and eventual lysis. FasL/Fas (CD95 ligand/CD95)–mediated cytolysis requires de novo protein synthesis of FasL by the CTL and the presence of the death receptor Fas on the target cell to induce apoptosis. Using a CD8+ CTL clone that kills via both the perforin/granule exocytosis and FasL/Fas mechanisms, and a clone that kills via the FasL/Fas mechanism only, we have examined the requirement of intra- and extracellular Ca2+ in TCR-triggered cytolytic effector function. These two clones, a panel of Ca2+ antagonists, and agonists were used to determine that a large biphasic increase in intracellular calcium concentration, characterized by release of Ca2+ from intracellular stores followed by a sustained influx of extracellular Ca2+, is required for perforin/granule exocytosis. Only the sustained influx of extracellular Ca2+ is required for FasL induction and killing. Thapsigargin, at low concentrations, induces this small but sustained increase in [Ca2+]i and selectively induces FasL/Fas-mediated cytolysis but not granule exocytosis. These results further define the role of Ca2+ in perforin and FasL/Fas killing and demonstrate that differential Ca2+ signaling can modulate T cell effector functions