CD146 is a novel marker for highly tumorigenic cells and a potential therapeutic target in malignant rhabdoid tumor.

Malignant rhabdoid tumor (MRT) is a rare, highly aggressive pediatric malignancy that primarily develops during infancy and early childhood. Despite the existing standard of intensive multimodal therapy, the prognosis of patients with MRT is dismal; therefore, a greater understanding of the biology of this disease is required to establish novel therapies. In this study, we identified a highly tumorigenic sub-population in MRT, based on the expression of CD146 (also known as melanoma cell adhesion molecule), a cell adhesion molecule expressed by neural crest cells and various derivatives. CD146+ cells isolated from four MRT cell lines by cell sorting exhibited enhanced self-renewal and invasive potential in vitro. In a xenograft model using immunodeficient NOD/Shi-scid IL-2Rγ-null mice, purified CD146+ cells obtained from MRT cell lines or a primary tumor exhibited the exclusive ability to form tumors in vivo. Blocking of CD146-related mechanisms, either by short hairpin RNA knockdown or treatment with a polyclonal antibody against CD146, effectively suppressed tumor growth of MRT cells both in vitro and in vivo via induction of apoptosis by inactivating Akt. Furthermore, CD146 positivity in immunohistological analysis of 11 MRT patient samples was associated with poor patient outcomes. These results suggest that CD146 defines a distinct sub-population in MRT with high tumorigenic capacity and that this marker represents a promising therapeutic target.

Post-Fukushima radiation education for public health nursing students: a case study.

BACKGROUND: The recent Fukushima Nuclear Power Plant accident was one of more than 200 serious nuclear/radiation incidents (accidents and disasters) that occurred worldwide since 1945. The current Fukushima disaster is in the recovery phase with the decreasing levels of radiation in the environment. However, fears and stigma related to the perceived risk of radiation exposure persist among the general population. INTRODUCTION: To improve on students' preparedness for social and public health challenges after a radiation incidence, radiation education was provided for undergraduate public health nursing students. AIM: This case study reports the development and implementation of the first class of radiation education in public health nursing, as well as students' reflections on their class experience. METHODS: We included a 90-min radiation class in an undergraduate public health nursing course in Tokyo, Japan. Lectures/discussion on technical and environmental aspects provided the minimally essential content for basic radiation knowledge. After class, all the 65 students were invited to freely write their reflections on the class. With their consent, 61 students' anonymous written accounts were qualitatively analysed. RESULTS: Five themes emerged: awareness of ignorance about radiation, problems produced by the mass media, becoming knowledgeable about radiation, public health nurses' role, and trustful and enjoyable lecture. DISCUSSION: The class inspired students to consider social, psychological and relational aspects of knowing and not knowing about radiation and their future professional role. CONCLUSION AND IMPLICATIONS FOR NURSING: Once radiation is taught at school, nursing students will emerge as professionals with the belief that radiation is within their professional purview. Education is key to disaster prevention, preparation, response and recovery. Given the ubiquitous nature of health challenges after a radiation incident, radiation education is indispensable for nursing students worldwide.

Solitary myofibroma of the lumbar vertebra: adult case.

We present the first known adult case of solitary myofibroma of bone, which affected a lumbar vertebra in a 33-year-old male. Radiography identified a purely lytic lesion with a sclerotic rim in the right pedicle of L1. CT showed an expansile lytic lesion with a sclerotic rim. MRI of the lesion revealed an isointense signal on T1-weighted images, an inhomogeneously hyperintense signal on T2-weighted images, and marked enhancement with gadolinium. Pathological study showed a mixed picture of nodular proliferation of spindle-shaped myoid cells and hemangiopericytomatous proliferation of short spindle/small round cells. The tumor cells were immunoreactive for smooth muscle actin and immunonegative for desmin. This case of solitary myofibroma of bone is exceptionally rare because of its occurrence in an adult older than 20 years of age and its location at an extra-craniofacial site.

The telling and knowing of dying: philosophical bases for hospice care in Japan.

This article describes the historical and cultural factors that led to the modern definition of hospice in the United States and Japan. It also raises questions about the import of western hospice philosophy into Japan by examining some traditional values and cultural norms of behaviour surrounding dying in that country. The ethics of disclosure and open discussion, as a prerequisite to and foundational value in western hospice care, differs in Japan. If traditional Japanese values remain strong, expectations regarding hospice care based on this western ethics may be misplaced. This report ends with a suggestion of replacing the western imported hospice philosophy with an (as yet not explicated) Japanese philosophical ethic of the good.

The right-to-die and the duty-to-die: perceptions of nurses in the West and in Japan.

The right-to-die means that the terminally ill person has the right to refuse further treatment. The duty-to-die means that terminally ill persons feel that they have no choice and must refuse treatment because of social factors such as family burden or financial cost to society. This article describes the perceptions of a sample of 72 Japanese nurses and 71 counterparts from the West. The right-to-die received support from all of the western and from a majority of the Japanese sample. However, the duty-to-die received weaker support from the West and rather strong disagreement from Japan. Themes emerged in the two groups from a content analysis of their reactions to the major concepts of right-to-die and duty-to-die, and these included autonomy, values surrounding life, justice, family as a possible coercive agent and family as a partner in decision making. Differences and similarities across the cultures in the meaning of these themes were explored.

Extraskeletal osteosarcoma arising in myositis ossificans.

A 53-year-old woman had extraskeletal osteosarcoma that developed from a soft tissue bony mass present on the volar aspect of the left wrist for 4 years. Initially, the bony mass was soft and movable, but during the first year it became hard and fixed. The patient had no history of trauma. Because the lesion did not grow or cause any symptoms, the patient did not come to the hospital until 4 years after she first noticed the lesion. Radiologically, the bony mass had features characteristic of mature myositis ossificans, showing "eggshell" ossification. A nonmineralized soft tissue mass occurred between the surface of the radius and the bony shell. Histologically, a high-grade osteosarcoma was present between the surface of the radius and the well-differentiated bone tissue, which included fatty and hematopoietic marrow. All the findings indicated that our patient had an extremely rare case of malignant transformation of myositis ossificans.

Generation and characterization of a mammalian cell line continuously expressing Japanese encephalitis virus subviral particles.

We have generated a cell line (F cells) producing a secreted form of Japanese encephalitis virus (JEV) subviral particle (extracellular particles [EPs]) that contains the JEV envelope glycoprotein (E) and a precursor (prM) of the virion membrane protein (M). The F cells were engineered to synthesize these JEV products from a cDNA encoding a mutated (furin proteinase resistant) form of prM, since stable cell lines expressing E and the authentic form of prM could not be obtained, due (in part) to the cell-fusing ability of EPs containing E and M. Our biochemical alteration of the prM protein was critical for the successful production of EP-producing cell lines. EPs produced by F cells share the biochemical properties of empty viral particles produced by JEV-infected cells, except that the F-cell EPs lack hemagglutinating activity and M. F-cell EPs were recognized by a panel of monoclonal antibodies to E, and EPs were shown to be useful as vaccine candidates in mice and as diagnostic reagents in evaluating human immune responses to JE vaccination. The amounts of E antigen released into the culture fluid of F cells were similar to those found in virion fractions of JEV-infected cell culture fluids or JEV-infected weanling mouse brains (the current source of antigen used to produce human vaccines for JE). Thus, the F-cell line would appear to be a useful source of antigen for JE vaccines and diagnostics.

Safety and immunogenicity of NYVAC-JEV and ALVAC-JEV attenuated recombinant Japanese encephalitis virus--poxvirus vaccines in vaccinia-nonimmune and vaccinia-immune humans.

A controlled, randomized, double-blind clinical trial evaluated whether two attenuated recombinant poxviruses with identical Japanese encephalitis virus (JEV) gene insertions, NYVAC-JEV and ALVAC-JEV, were safe and immunogenic in volunteers. Groups of 10 volunteers distinguished by vaccinia immune status received two doses of each vaccine. The vaccines appeared to be equally safe and well tolerated in volunteers, but more reactogenic than licensed formalin-inactivated JE and placebo vaccines given as controls. NYVAC-JEV and ALVAC-JEV vaccine recipients had frequent occurrence of local warmth, erythema, tenderness, and/or arm pain after vaccination. There was no apparent effect of vaccinia immune status on frequency or magnitude of local and systemic reactions. NYVAC-JEV elicited antibody responses to JEV antigens in recipients but ALVAC-JEV vaccine poorly induced antibody responses. However, NYVAC-JEV vaccine induced neutralizing antibody responses only in vaccinia-nonimmune recipients while vaccinia-immune volunteers failed to develop protective antibodies (5/5 vs. 0/5 seroconversion, p<0.01). These data suggest that preexisting immunity to poxvirus vector may suppress antibody responses to recombinant gene products.

Identification of enzymes responsible for rifalazil metabolism in human liver microsomes.

1. The major metabolites of rifalazil in human are 25-deacetyl-rifalazil and 32-hydroxy-rifalazil. Biotransformation to these metabolites in pooled human liver microsomes, cytosol and supernatant 9000g (S9) fractions was studied, and the enzymes responsible for rifalazil metabolism were identified using inhibitors of esterases and cytochromes P450 (CYP). 2. The 25-deacetylation and 32-hydroxylation of rifalazil occurred in incubations with microsomes or S9 but not with cytosol, indicating that both the enzymes responsible for rifalazil metabolism were microsomal. Km and Vmax of the rifalazil-25-deacetylation in microsomes were 6.5 microM and 11.9 pmol/min/mg with NADPH, and 2.6 microM and 6.0 pmol/min/mg without NADPH, indicating that, although rifalazil-25-deacetylation did not require NADPH, NADPH activated it. Rifalazil-32-hydroxylation was NADPH dependent, and its Km and Vmax were 3.3 microM and 11.0 pmol/min/mg respectively. 3. Rifalazil-25-deacetylation in microsomes was completely inhibited by diisopropyl fluorophosphate, diethyl p-nitrophenyl phosphate and eserine, but not by p-chloromercuribenzoate or 5,5'-dithio-bis(2-nitrobenzoic acid), indicating that the enzyme responsible for the rifalazil-25-deacetylation is a B-esterase. 4. Rifalazil-32-hydroxylation in microsomes was completely inhibited by CYP3A4-specific inhibitors (fluconazole, ketoconazole, miconazole, troleandomycin) and drugs metabolized by CYP3A4 such as cyclosporin A and clarithromycin, indicating that the enzyme responsible for the rifalazil-32-hydroxylation is CYP3A4.

Definition of an epitope on Japanese encephalitis virus (JEV) envelope protein recognized by JEV-specific murine CD8+ cytotoxic T lymphocytes.

We defined an epitope on the Japanese encephalitis virus (JEV) envelope (E) protein recognized by CD8+ cytotoxic T lymphocytes (CTLs). CTLs induced in JEV-infected BALB/c (H-2d) mice recognized E and/or premembrane (PrM) proteins, while CTLs in C57BL/6J (H-2b) and C3H/HeJ (H-2k) mice did not. JEV-specific CTLs had a phenotype of CD3+ CD4- CD8+. Twenty-four 9-amino acid (a.a.) peptides, which had binding motifs for H-2Kd, H-2Ld or H-2Dd, were synthesized according to the amino acid sequences of PrM and E proteins. CTLs induced by JEV infection recognized only the peptide K-3. Immunization of BALB/c mice with only a group of peptides including K-3 induced CTLs which recognized the homologous K-3 peptide, while immunization with other peptides did not. The peptide K-3 had a binding motif for H-2Kd. This is consistent with the finding that JEV-specific CTLs in BALB/c mice was H-2Kd-restricted. These results indicate that the epitope recognized by CTLs in BALB/c mice is located between a.a. 60 and 68 on the E protein, corresponding to an a.a. sequence of CYHASVTDI.

End-of-life ethical issues in Japan.

This article discusses the end-of-life ethical issues of disclosure of information to the terminally ill patient, informed consent, and overtreatment for the elderly people of Japan. These issues are placed within the cultural context of Japan, which is discussing and developing ways to deal with them. The ethical problems that nurses confront when they work clinically with elderly patients also are outlined. No consensus has been reached about the ethics of health care in Japan.

Japanese encephalitis DNA vaccine candidates expressing premembrane and envelope genes induce virus-specific memory B cells and long-lasting antibodies in swine.

Swine are an important amplifier of Japanese encephalitis (JE) virus in the paradomestic environment. In this study, two JE DNA vaccine candidates were evaluated for immunogenicity in swine. Both vaccine plasmids encode a cassette consisting of the signal of premembrane (prM), prM, and envelope (E) coding regions of JE virus. One plasmid, designated pcJEME, is based on a commercial vector (pcDNA3), whereas the other plasmid, designated pNJEME, is based on a vector (pNGVL4a) designed to address some of the safety concerns of DNA vaccine use. No differences were detected in the immunogenicity of these two plasmids in mice or swine. Swine immunized with the DNA vaccines at a dose of 100 to 450 microgram at an interval of 3 weeks developed neutralizing and hemagglutination-inhibitory (HAI) antibody titers of 1:40 to 1:160 at 1 week after the second immunization. However, swine administered two doses of a commercial JE vaccine (formalin-inactivated virus preparation; JEVAX-A) developed low (1:10) or undetectable antibody responses after their boost. Interestingly, serum antibody titers elicited by DNA vaccines in swine were higher than those detected in mice. Eight days after boosting with viral antigen (JEVAX-A) to detect an anamnestic response, swine immunized two times with the DNA vaccine showed a >100-fold elevation in HAI titer, indicating a strong recall of antibody response. Swine maintained detectable levels of HAI antibody for at least 245 days after two immunizations with a DNA vaccine. These results indicate that these DNA vaccines are able to induce virus-specific memory B cells and long-lasting antibodies in swine, which were of higher levels than those obtained with a commercial formalin-inactivated JE vaccine.

A DNA vaccine expressing dengue type 2 virus premembrane and envelope genes induces neutralizing antibody and memory B cells in mice.

A dengue DNA vaccine candidate was developed and evaluated for immunogenicity in mice. The vaccine, designated pcD2ME, is a pcDNA3-based plasmid encoding the signal sequence of premembrane (prM), prM and envelope (E) genes of the New Guinea C strain of dengue type 2 virus. CHO-K1 cells transfected with pcD2ME expressed prM and E as determined by immunochemical staining with monoclonal antibodies. BALB/c mice inoculated intramuscularly with 100 microg of pcD2ME two or three times at an interval of 2 weeks developed a low level of neutralizing antibody (1:10 at a 90% plaque reduction). Immunization twice with 10 microg or 1 microg of pcD2ME or three times with 100 microg of pcDNA3 did not induce detectable levels of neutralizing antibody. Mice immunized two or three times with 100 microg of pcD2ME raised neutralizing antibody titers to 1:40 or greater on days 4 and 8 after challenge with 3x10(5) plaque forming units (PFU) of the New Guinea C strain of dengue type 2 virus, showing strong anamnestic responses to the challenge. In contrast, mice immunized two or three times with 100 microg of pcDNA3 developed no detectable neutralizing antibody on days 4 and 8 after challenge. These results indicate that immunization with pcD2ME induces neutralizing antibody and dengue type 2 virus-responsive memory B cells in mice.

Contamination of passenger trains with Dermatophagoides (Acari: Pyroglyphidae) mite antigen in Japan.

Passenger trains were surveyed for contamination with Dermatophagoides farinae Hughes and Dermatophagoides pteronyssinus (Trouesart) mites in Japan. A total of 492 dust samples were collected from upholstered seats in six commuter trains, one long-distance express train and three night trains in October, 1996 and January, April, and July, 1997. Mite antigen levels contained in fine dust fractions of these samples were measured by an enzyme-linked immunosorbent assay. Most samples obtained from commuter trains showed relatively high mite antigen levels of > 10 microgm(-2) (corresponding to > 100 mites). Express and night trains showed lower antigen levels per square meter, but higher mite antigen levels per gram of fine dust than commuter trains, indicating relatively high mite antigen densities. Seasonal comparisons indicated that commuter trains showed the highest mean antigen level per square meter in winter (January), whereas the highest antigen level per gram of fine dust was observed in summer (July) in express and night trains.

Prevalence of antibodies to Toxoplasma gondii among urban and rural residents in the Philippines.

A survey of antibodies to Toxoplasma gondii was carried out among residents in urban (Metro Manila) and rural (Mindoro and Leyte) areas in the Philippines. A total of 1,173 serum samples were examined for Toxoplasma antibody by an ELISA method. The overall seropositivity was 11.1% (n=904, 12.4% in males, 10.0% in females) in Metro Manila, 61.2% (n=152, 63.3% in males, 53.1% in females) in Mindoro, and 30.1% (n=113, 34.3% in males, 22.5% in females) in Leyte, indicating significantly higher (p<0.001) seropositivities in rural than urban settings. No significant differences in seropositivities were observed between males and females. In each group, seropositivity tended to increase with age of the subjects.

Gastric cancer presenting with extremely rapid growth: unprecedented morphologic change in a short time and endoscopic estimation of its doubling time.

We encountered a case of gastric cancer that was initially detected as a deep hemorrhagic ulcer without surrounding irregular elevation, followed by rapid protrusion in less than 1 month. Using endoscopic images in the follow-up study, we estimated the doubling time (DT) of this unusual tumor as 9.2 days. Since the doubling time of gastric cancer is generally fairly long due to exfoliation of many cancer cells into the gastric lumen, this cancer presented with extremely rapid growth. Besides, this case reinforces that follow-up study is important in terms of clinical management of ulcerative lesions.

Isolation and identification of major metabolites of rifalazil in mouse and human.

1. Three metabolites of rifalazil have been isolated from dog urine and identified as 25-deacetyl-rifalazil, 30-hydroxy-rifalazil and 25-deacetyl-30-hydroxy-rifalazil. In the current study major metabolites of rifalazil in mouse and human were isolated and identified, and their antimicrobial activities determined. 2. Urinary excretion of rifalazil and its metabolites in six mouse strains, CD-1 (ICR), BALB/c, C57BL/6, C3H/He, DBA/2 and CBA/J, was examined. Two major metabolites were detected in mouse urine obtained after several oral doses, and the proportion of rifalazil metabolites against total urinary excretion varied over a 2-fold range (4.8-8.7%) in the different mouse strains. 3. One of two major metabolites in mouse urine was 25-deacetyl-rifalazil and the other was unknown: it was isolated from mouse urine and identified by ms and 1H- and 13C-nmr as 32-hydroxy-rifalazil. 4. In human, two major metabolites of rifalazil were detected in urine obtained after administration of a single oral dose. These metabolites were also produced by incubation of rifalazil with pooled human liver microsomes, and identified by lc/ms and lc/ms/ms as 25-deacetyl-rifalazil and 32-hydroxy-rifalazil. 5. The antimicrobial activities of 32-hydroxy-rifalazil against gram-positive bacteria and mycobacteria were similar with those of the parent compound.

The anamnestic neutralizing antibody response is critical for protection of mice from challenge following vaccination with a plasmid encoding the Japanese encephalitis virus premembrane and envelope genes.

For Japanese encephalitis (JE), we previously reported that recombinant vaccine-induced protection from disease does not prevent challenge virus replication in mice. Moreover, DNA vaccines for JE can provide protection from high challenge doses in the absence of detectable prechallenge neutralizing antibodies. In the present study, we evaluated the role of postchallenge immune responses in determining the outcome of JE virus infection, using mice immunized with a plasmid, pcDNA3JEME, encoding the JE virus premembrane (prM) and envelope (E) coding regions. In the first experiment, 10 mice were vaccinated once (five animals) or twice (remainder) with 100 micrograms of pcDNA3JEME. All of these mice showed low (6 of 10) or undetectable (4 of 10) levels of neutralizing antibodies. Interestingly, eight of these animals showed a rapid rise in neutralizing antibody following challenge with 10,000 50% lethal doses of JE virus and survived for 21 days, whereas only one of the two remaining animals survived. No unimmunized animals exhibited a rise of neutralizing antibody or survived challenge. Levels of JE virus-specific immunoglobulin M class antibodies were elevated following challenge in half of the unimmunized mice and in the single pcDNA3JEME-immunized mouse that died. In the second experiment, JE virus-specific primary cytotoxic T-lymphocyte (CTL) activity was detected in BALB/c mice immunized once with 100 micrograms of pcDNA3JEME 4 days after challenge, indicating a strong postchallenge recall of CTLs. In the third experiment, evaluation of induction of CTLs and antibody activity by plasmids containing portions of the prM/E cassette demonstrated that induction of CTL responses alone were not sufficient to prevent death. Finally, we showed that antibody obtained from pcDNA3JEME-immunized mice 4 days following challenge could partially protect recipient mice from lethal challenge. Taken together, these results indicate that neutralizing antibody produced following challenge provides the critical protective component in pcDNA3JEME-vaccinated mice.

Long-term care for the elderly in Japan.

This article, using data from the first author's research, presents selected issues in long-term care (LTC) of the elderly in Japan. A brief discussion of historical and cultural factors frame the current realities of LTC. These realities include the vast numbers of elderly people in Japan, changing definitions of the relationship of the individual to the group, and enactment of the new Care Insurance Law for the Elderly to be implemented in the year 2000. Some of the work underway for this implementation is detailed.

Japanese nurses' perceptions about disclosure of information at the patients' end of life.

Information disclosure at the end of life is one of the most debated ethical issues in Japan. This paper, using data from a larger questionnaire survey in which 147 Japanese nurses participated, describes nurses' perceptions about this issue. The nurses perceived that non-disclosure of impending death information to patients was the norm in Japan due to its traditional values. This non-disclosure of information has various impacts on clinical nurses. Tension was evident between the Japanese traditional concepts supported by some nurses and the shift toward Western cultural and ethical values supported by other nurses. The nurses confronted uninformed patients who became suspicious, isolated, angry, or died unprepared for their life's ending. The nurses were placed in the middle between the patient and the family, as nurses became keepers of family secrets, or between nurses' ethical obligations to the patient and those of the institution. The nurses had the belief that at the patient's end of life, it is important to change health professionals' attitude from curing to caring.