Teaching "medical interview and physical examination" from the very beginning of medical school and using "escape rooms" during the final assessment: achievements and educational impact in Japan.

BACKGROUND: There is no consensus regarding the best time to teach two fundamental pillars of clinical medicine: medical interview and physical examination. We investigated the impacts of teaching the course "Medical Interview and Physical Examination" in Japan from the very beginning of medical school. In addition, we also evaluated the educational value of using "Escape Rooms", a series of timed, game-based scenarios using simulators, as a part of the final assessment of the course. METHODS: At the end of the course, the interview capabilities of 140 first year medical students at International University of Health and Welfare (Japan) were assessed by physicians who acted as simulated patients. Physical examination skills were assessed using the "Escape Room" team task method. Students also self-assessed their confidence in their physical examination skills pre and post "Escape Rooms." A day prior to the final assessment, students completed an anonymous course evaluation. RESULTS: The average global rating of the students' medical interview skills using a rating scale from 1 to 6 (1-fail 6-outstanding, no different from practicing junior physician's level) was 4.6. Twenty-two students scored the highest mark of 6. An average of 89% of "Escape Room" teams finished all the physical examination tasks correctly within the allotted time. All teams that could not finish in time completed all tasks correctly when given an additional 3 to 5 min. Students' self-assessed confidence in their physical examination skills increased from 49 to 73 (out of 100) pre and post "Escape Rooms." In the course evaluation questionnaire, 99% of students answered "this course enhanced their motivation" (response rate 89%) and 99% also answered "this course was interesting and useful" (response rate 86%). CONCLUSIONS: This descriptive study analyzing both quantitative and qualitative data showed that the course not only achieved the intended objectives of successfully conducting comprehensive medical interview and basic physical examination skills, but also enhanced student motivation. "Escape Rooms", used for the course assessment, in itself enhanced students' self-perceived physical examination skills and had an added educational value.

Rapid and real-time detection of hepatitis A virus by reverse transcription loop-mediated isothermal amplification assay.

A one-step, single tube, real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detecting sequences of the untranslated region in the genome of hepatitis A virus (HAV). The RT-LAMP assay reported in this study was very simple and rapid; the HAV-specific amplification was obtained in 50 min under isothermal conditions at 62.5 degrees C by employing a set of seven primers. The RNAs of three cell-adapted HAV strains belonging to different subgenotypes (IA, IB and IIIB) were equally well amplified. The detection limits of the RT-LAMP assay for these HAV strains were 0.4-0.8 focus forming units (FFU)/reaction. The results of the calibration using the WHO international standard indicated that the RT-LAMP assay had similar sensitivity to the conventional RT-PCR method. A comparison of the results from the RT-LAMP and the LightCycler PCR assay using clinical samples in feces revealed that the findings were similar between the two methods. Although several genotypes remain to be tested, it is concluded that the new real-time RT-LAMP assay is very suitable for detection and quantitation of most prevalent genotypes of HAV in diagnostic laboratories.

Characteristics and Gender Differences in the Medical Interview Skills of Japanese Medical Students.

Objective To examine the characteristics of the communication skills of medical students, we observed their performance during introductory medical interview training with simulated patients (SPs). Methods The subjects of the present study included fifth-year medical students (male, n=180, female, n=99) who were undergoing clinical training in Japan from 2012 to 2014. Each student was assigned to one of four 10-minute clinical scenarios, which was conducted with an SP. Three or four teachers observed and assessed the performance of each of the students. The overall performance was rated on a 10-point scale, and nine basic communication skills that were common to each of the scenarios were rated using a four-point scale. The students also assessed their own performance on these items. The SPs assessed the students' performance from a patient's perspective on four items. Results There were significant correlations between the teacher and student scores. However, the students tended to score themselves significantly lower than the teachers. The female students were rated significantly higher by the teachers on the following four items; 'eye contact and appropriate attitude,' 'nodding and back-channeling,' 'giving empathic verbal responses,' and 'acquisition of patient's psychosocial information.' However, the self-assessments of the female students were only significantly higher than the male students in one item, 'acquisition of patient's psychosocial information.' In contrast, self-assessments of the male students were significantly higher in two items; none of their items was scored higher by the teachers. There was no significant gender difference in the assessments made by the SPs. Conclusion There were significant gender differences in the communication skills of the medical students during introductory training, suggesting the possibility that there were gender-specific traits and gender-based differences in the students' degrees of readiness.

CpG oligonucleotides enhance the tumor antigen-specific immune response of a granulocyte macrophage colony-stimulating factor-based vaccine strategy in neuroblastoma.

Granulocyte macrophage colony-stimulating factor (GM-CSF)-transduced autologous tumor cells form the basis of many immunotherapeutic strategies. We tested whether combining this approach with T-helper 1 (Th-1)-like immunostimulatory CpG oligodeoxynucleotides (CpG ODNs) would improve therapeutic efficacy in an established model of murine neuroblastoma. The weakly immunogenic Neuro-2a cell line was used in syngeneic A/J mice. CpG 1826 was tested for its antitumor effect alone and as an adjuvant to Neuro-2a cells retrovirally transduced to express murine GM-CSF (GM/Neuro-2a). Three days after wild-type (WT) tumor cell inoculation, mice in different groups were s.c. vaccinated in the opposite leg with combinations of WT neuro2a, irradiated (15 Gy) WT or GM/Neuro-2a transfectants with or without CpG 1826 (200 micro g). To test for the induction of memory responses, mice that rejected their tumor were rechallenged with WT Neuro-2a (1 x 10(6)) 7 weeks after vaccination. All of the mice in the control (unvaccinated) group died within 3 weeks after Neuro-2a inoculation. Most of the vaccinated groups had only minimal-to-modest antitumor responses, and the mice succumbed to tumor. Tumor growth was remarkably inhibited in the group of mice that received irradiated GM/Neuro-2a plus CpG and four (50%) of eight mice in this group survived tumor free. Tumor-free mice were resistant to further WT tumor cell challenge, indicating a memory response. Mechanistic studies showed that CpG alone induced a favorable Th-1-like cytokine immune response and vaccine-induced tumor cell killing was dependent on both CD4 and CD8 T cells that killed tumor cell targets by apoptosis. These results demonstrate that CpG ODNs enhanced the antitumor effect of irradiated GM-CSF secreting Neuro-2a cells. This vaccine strategy elicits a potent tumor antigen-specific immune response against established murine neuroblastoma and generates systemic neuroblastoma-specific immunity.

Isolation and autoxidation profile of fish myoglobin from hoki (Macruronus magellanicus).

Myoglobin is known to be present exclusively in cardiac and red skeletal muscles, but not in white skeletal muscles. Thus, to date, only a few studies on myoglobin from fish species with white flesh have been reported. For comparative examination, we directly isolated myoglobin from cardiac muscle of hoki (Macruronus magellanicus), one of the most important commercial fish species with white muscle. The ferrous myoglobin was separated from its ferric met-form by anion exchange column chromatography. The absorption spectra of hoki myoglobin were similar to those of bigeye tuna skeletal myoglobin, in both oxy- and met-forms. However, hoki oxymyoglobin was found to be susceptible to autoxidation in 0.1 M buffer (pH 7.2) at 25°C, with its rate being more than 3 times higher than that of bigeye tuna oxymyoglobin.

Surgical excision combined with autologous whole tumor cell vaccination is an effective therapy for murine neuroblastoma.

BACKGROUND: Although a whole tumor cell vaccine strategy based on the synergistic action of granulocyte macrophage-colony-stimulating factor (GM-CSF) transduced tumor cells and CpG oligodeoxynucleotides induces potent tumor-specific immunity, such therapy is not curative in the face of large established tumors. PURPOSE: The primary goal of this study was to determine if combining surgical resection of the tumor with whole tumor cell vaccination is an effective therapeutic strategy for established neuroblastoma. We also wished to determine if the phenotype of the immune response generated by this vaccine strategy was altered by the presence of bulky established tumor. METHODS: The murine neuroblastoma model was used in which syngeneic Neuro-2a was grown subcutaneously in A/J mouse. The whole tumor cell vaccine consisted of irradiated Neuro-2a as the vehicle for tumor antigens admixed with GM-CSF and CpG oligonucleotides (100 microg). RESULTS: In the presence of large tumors, dendritic cells were effectively activated by the vaccine, but secretion of intereferon-gamma from vaccinated splenocytes in response to antigen stimulation was suppressed. The tumor's inhibitory effect on interferon-gamma production from vaccinated splenocytes was reversed after resection of the primary tumor. The use of prepared GM-CSF-secreting bystander cells simplified production of an autologous whole tumor cell vaccine that was remarkably effective in curing large tumors when combined with tumor excision. CONCLUSIONS: These findings suggest that at least part of the immunosuppressive effects of the cancer can be reversed after surgical excision of the primary tumor. Thus, in the context of minimal residual disease, this dendritic cell-targeted immunotherapeutic approach may prove effective for the treatment of neuroblastoma.

Yeast flavohemoglobin from Candida norvegensis. Its structural, spectral, and stability properties.

Flavohemoglobin was isolated directly from the yeast Candida norvegensis and studied on its structural, spectral, and stability properties. In Candida flavohemoglobin, the 155 N-terminal residues make a heme-containing domain, while the remaining 234 C-terminal residues serve as a FAD-containing reductase domain. A pair of His-95 and Gln-63 was assigned to the proximal and distal residues, respectively. In purification procedure FAD was partially dissociated on a Butyl-Toyopearl column, so that FAD-lacking flavohemoglobin was also obtainable. In this ferric species, the Soret and charge-transfer bands were all characteristic of a penta-coordinate form. Compared with the recombinant heme domain expressed in Escherichia coli, we have measured the autoxidation rate over a wide pH range. The resulting pH dependence curves were then analyzed in terms of a nucleophilic displacement mechanism. As a result, the heme domain was found to be extremely susceptible to autoxidation, its rate being more than 100 times higher than that of sperm whale MbO2. However, this inherently high oxidation rate was dramatically suppressed in Candida flavohemoglobin to an extent almost comparable to the stability of mammalian myoglobins. These new findings lead us to conclude that Candida flavohemoglobin, differently from bacterial flavohemoglobins, can serve as an oxygen storage protein in aerobic conditions.