Development of a wearable computer system with a hands-free operation interface for the use of home health caregiver.

We previously developed a network-based medical care support system called the Hyper Hospital, a computer network with an interface that is dedicated to patient care. In this study, we developed a wearable information system that is designed so that a caregiver can obtain information and control various support devices within the home-care environment. In our system, the wearable computer itself consists of a computer network built into a jacket. Each required function is implemented by a dedicated small computer connected to the in-jacket network. A new function may easily be added to the system by connecting additional computers. A network comprising such a set of single-function computers becomes a highly efficient information system when applied to health care support.

Antitumor activity of a phenoxazine compound, 2-amino-4,4alpha-dihydro-4alpha,7-dimethyl-3H-phenoxazine-3-one against human B cell and T cell lymphoblastoid cell lines: induction of mixed types of cell death, apoptosis, and necrosis.

PURPOSE: We studied the antitumor activity of 2-amino-4,4alpha-dihydro-4alpha,7-dimethyl-3H-phenoxazine-3-one (Phx), which was synthesized by the reactions of 2-amino-5-methylphenol with bovine hemoglobin, on human B cell lymphoblastoid cell lines, P3HR-1 and Raji derived from African Burkitt's lymphoma, and the human T cell lymphoblastoid cell line Molt-4. We also studied whether Phx might cause apoptosis and necrosis in these cells. METHODS: We evaluated cell viability and apoptosis and necrosis of the cells in the presence of Phx, by using agarose gel electrophoresis, flow cytometry, and fluorescence microscopy. RESULTS: Phx suppressed the viability of P3HR-1, Raji, and Molt-4 cells, though the suppression patterns were different, i.e., Phx suppressed the viability of P3HR-1, Raji, and Molt-4 cells at higher concentrations, while the drug enhanced the viability of Raji cells, but not those of P3HR-1 and Molt-4 cells at lower concentrations. To investigate which type of cell death - apoptosis or necrosis - is induced by Phx, induction of DNA ladder, phosphatidylserine externalization, and propidium iodide-permeable cells were examined in Phx-treated cells. Although Phx did not induce DNA ladder formation, it induced the phosphatidylserine externalization and propidium iodide-permeable cells, suggesting that Phx caused a mixed type of cell death, both apoptosis and necrosis. The population of early stage apoptotic cells was dominant in Raji cells, and that of the late stage apoptotic/necrotic cells was dominant in Molt-4 cells after 72-h treatment with Phx. The population of the early stage apoptotic cells and the late stage apoptotic/necrotic cells was almost equal in P3HR-1 cells in the presence of Phx, though the population of both types of cells increased with time. The nuclear morphological analysis of Phx-treated Raji, P3HR-1, and Molt-4 cells also showed that Phx induces apoptosis. CONCLUSIONS: The present results suggest that Phx shows antitumor activity against human B cell-derived and T cell-derived lymphoblastoid cell lines, in vitro, causing apoptosis and necrosis.

Possible involvement of EBV-mediated alpha-fodrin cleavage for organ-specific autoantigen in Sjogren's syndrome.

A cleavage product of alpha-fodrin may be an important organ-specific autoantigen in the pathogenesis of Sjogren's syndrome (SS), but the mechanisms of alpha-fodrin cleavage remain unclear. Since EBV has been implicated in the pathogenesis of SS, we determined whether EBV activation could induce the SS-specific 120-kDa autoantigen alpha-fodrin. ZEBRA mRNA expression, a marker for activation of the lytic cycle of EBV, was found in the salivary gland tissues from SS patients, but not in those from control individuals. ZEBRA-expressing lymphoid cells were also found in the SS glands in double-stained immunohistochemistry. Furthermore, a significant link between production of Abs against 120-kDa alpha-fodrin and reactivated EBV Ag was found in sera from patients with SS, but not in those from control individuals. EBV-activated lymphoid cells showed specific alpha-fodrin cleavage to the expected 120-kDa fragments in vitro. Pretreatment with caspase inhibitors inhibited cleavage of alpha-fodrin. Thus, an increase in apoptotic protease activities induced by EBV reactivation may be involved in the progression of alpha-fodrin proteolysis in the development of SS.

Infection of human lung fibroblasts with Epstein-Barr virus causes increased IL-1beta and bFGF production.

An association between Epstein-Barr virus (EBV) infection and fibroblast proliferation in the interstitial spaces of the lung has been suggested in idiopathic interstitial pneumonia. In this study we show that EBV can infect human lung fibroblasts in vitro. A primary-cultured human lung fibroblast cell line, designated CCD-32Lu, expressed EBV nuclear antigen 1 after coculture with lethally irradiated EBV producing cells. The infection further induced CCD-32Lu cells to produce the fibrogenic cytokines basic fibroblast growth factor (bFGF) and interleukin-1beta. These findings indicate that lung fibroblasts may be a target for EBV infection and suggest that EBV may play a role in increased production of these cytokines and induce fibroblast proliferation in idiopathic interstitial pneumonia.

Detection of lymphocytes productively infected with Epstein-Barr virus in non-neoplastic tonsils.

Epstein-Barr virus (EBV) persists for life in the infected host. Little is known about EBV reactivation and regulation of virus persistence in healthy individuals. We examined tonsils of chronic tonsillitis patients to detect EBV transcripts, EBV genomes and lytic proteins. LMP1 transcripts were observed in 11 of 15 specimens and BZLF1 transcripts were detected in six. Multiple copies of EBV genome equivalents per cell, and ZEBRA- and viral capsid antigen-positive cells were also detected in tonsillar lymphocytes. These results indicate that EBV productively infected cells may survive in the face of immune surveillance in the tonsils. Thus, EBV replication may occur in tonsillar lymphocytes, and tonsillar lymphoid tissues may play a role in the maintenance of EBV load in vivo.

Activation of nuclear factor kappaB at the onset of ossification of the spinal ligaments.

We examined the correlation between the activation of nuclear factor kappaB (NFkappaB), stimulated by environmental factors involving cytokines and growth factors in ligament cells, and the onset of ossification of the spinal ligaments (OSL) or diffuse idiopathic skeletal hyperostosis (DISH). Aseptic samples were taken carefully from non-ossified sites during surgery (75 patients). We carried out preliminary hematoxylin and eosin and toluidine blue staining, using five portions of each specimen, and excluded samples containing chondrocytic, osteoblastic, or inflammatory cells (n = 25). We used specimens from the remaining 50 patients (35 men and 15 women, ranging in age from 45-81 years); average age, 59.5 years (18 nuchal ligament specimens, and 32 yellow ligament specimens). OSL or DISH had occurred in 25 patients, 20 patients were in the non-OSL group (8 with cervical spondylotic myelopathy, and 12 with lumbar canal stenosis), and the remaining 5 samples were collected from patients with injury. For culture study, we used portions of the 14 largest samples from the above 50 patients. We extracted nuclear proteins and cytoplasmic proteins from non-ossified spinal ligaments in 50 patients and detected p65RelA/NFkappaB by Western blotting. Tumor necrosis factor-alpha (TNF alpha), interleukin 1beta (IL-1beta), platelet-derived growth factor BB (PDGF-BB) and transforming growth factor-beta1 (TGF-beta1) in cytoplasm were quantified by enzyme-linked immunosorbent assays (ELISA). Cultured cells from the 14 samples were then stimulated with 10, 100, 250, or 500 ng/ml of recombinant human (rh)PDGF-B or TGFbeta1. A control experiment was performed without rhPDGF-BB or TGFbeta1 stimulation. Alkaline phosphatase (ALP) activity was standardized by the DNA content of the cells. The number of NFkappaB-positive samples was significantly higher in patients with OSL or DISH than in non-OSL patients. This tendency was obvious in the case of OSL or DISH with non-insulin-dependent diabetes mellitus (NIDDM). In OSL and in DISH patients, significantly greater amounts of PDGF-BB and TGFbeta1 were seen in ligament cells than in non-OSL patients (P < 0.05). There was a positive correlation between the detection of p65RelA/NFkappaB band and the content of PDGF-BB and TGFbeta1 in ligament cells (P < 0.05). ALP activity tended to be higher in cells in the OSL group not receiving any other treatment. Our results indicate the possibility that NFkappaB, stimulated by environmental factors involving PDGF-BB and TGFbeta1 in ligament cells, influences the osteoblastic differentiation of undifferentiated mesenchymal cells.

Behçet's disease and antibody titers to various heat-shock protein 60s.

It has been suggested that the 65 kDa heat-shock protein (HSP) of Streptococcus in recurrent aphthae within the oral cavity may be involved in the uveoretinitis of Behçet's disease, possibly through sensitization of the immune system. To investigate this possibility, we examined serum antibody titers for various members of the 60 kDa family of HSPs and their implications with regard to a role for HSP60s in Behçet's disease. We isolated HSP60 of Streptococcus pyogenes from the margin of oral aphthae in one Behçet's disease patient with severe uveoretinitis and the HSP60s of Yersinia enterocolitica, retinoblastoma cell line clone Y79, and bovine retinal extract and investigated the reaction of each of these HSP60s with 100-fold diluted serum samples from 20 Behçet's disease patients using anti-HSP60 antibody titers determined by ELISA. The anti- Streptococcus HSP60 antibody and anti-retinal HSP60 antibody titers of the 100-fold diluted serum samples from the Behçet's disease patients were both significantly higher than those of similarly diluted serum samples from healthy donors. The results of the ELISA antibody titer assay showed that, although the various HSP60s share a common basic antigenicity, they differed in reactivity to the anti-HSP60 antibodies in the sera of the Behçet's disease patients. The results indicate that subtle but significant differences exist in the antigenicity of the various HSP60s tested, all of which share a common basic antigenicity and are of approximately the same molecular weight, and suggest that an immuno-cross-reaction between retinal and streptococcal HSPs and a related autoimmune response may be involved in the development of Behçet's disease.

Endoscopic management of nasal glioma in infancy.

In the surgical treatment of nasal glioma, craniotomy has been recommended for excluding the possibility of intracranial extension of the lesion. We describe a case in whom intranasal glioma was successfully removed by endoscopic surgery without craniotomy at 4 months old. Intranasal endoscopic surgery is considered appropriate for the removal of intranasal glioma having no intracranial extension, since the procedure is less invasive and does not result in postoperative facial deformity. Intranasal endoscopic surgery is also proposed as the preferable procedure to craniotomy for excluding intracranial extension of intranasal glioma.

Possible involvement of IL-12 expression by Epstein-Barr virus in Sjögren syndrome.

AIM: To determine the correlation between interleukin 12 (IL-12) expression and Epstein-Barr virus (EBV) in Sjögren syndrome. METHODS: Indirect immunohistochemical technique, enzyme linked immunosorbent assay (ELISA), and immunoblot analysis were used to investigate IL-12 expression by EBV activation, using 13 surgical specimens and four B cell lines. RESULTS: Marked expression of IL-12 was found in the epithelial cells and the infiltrating B cells of salivary gland tissues from patients with Sjögren syndrome (six of 10 cases), but not in those from normal individuals (none of three cases). A striking topographic correlation between IL-12 and EBV was found. In addition, levels of IL-12 production by B cell lines were clearly enhanced by EBV activation in vitro. CONCLUSIONS: IL-12 expression closely reflects the intracellular event of EBV activation in Sjögren syndrome, and may contribute to the T helper cell type 1 (Th1) cytokine overexpression seen in this disease.

An Interlaboratory Validation Study of the Improved Transformation Assay Employing Balb/c 3T3 Cells: Results of a Collaborative Study on the Two-stage Cell Transformation Assay by the Non-genotoxic Carcinogen Study Group.

The Non-genotoxic Carcinogen Study Group of the Environmental Mutagen Society of Japan organised the first step of an interlaboratory validation study on an improved cell transformation assay employing Balb/c 3T3 A31-1-1 cells. Nineteen laboratories participated in this study. The modified transformation assay was evaluated for its responsiveness, its interlaboratory reproducibility and its transferability. In this study, a mixture of Dulbecco's modified Eagle's medium and nutrient mixture F12, supplemented with insulin-transferrin-ethanolamine-sodium selenite and 2% fetal bovine serum (FBS) was used during the period of expression of transformed foci, intead of the usual minimum essential medium with 10% FBS. 20-Methylcholanthrene (MCA) and 12-O-tetradecanoylphorbol-13-acetate (TPA) were selected as a prototype initiator and a tumour promoter, respectively. Two series of experiments were conducted. In the first series, the transformation activity of MCA was examined at various concentrations. In the absence of the promoting treatment with TPA, exposure to MCA only weakly induced transformed foci. In the presence of 0.1µg/ml TPA, all laboratories observed significant dose-dependent increases in the number of transformed foci with increasing MCA concentrations. In the second series of experiments, various concentrations of TPA were tested. In the absence of initiating treatment with MCA, exposure to TPA weakly induced transformed foci in about half of the laboratories. In the presence of 0.2µg/ml MCA, all the laboratories observed significant dose-dependent increases in the number of transformed foci with increasing TPA concentrations. The results from this study support the usefulness of this modified two-stage transformation assay with Balb/c 3T3 cells.

Optical and magneto-optical properties of copper-nickel compound metal island films.

The optical properties of copper Cu-Ni compound metal island (CMI) films composed of nanoclusters of an alloy of Cu and Ni have been experimentally investigated. The spectral characteristics of the Cu-Ni CMI films are intermediate between those of conventional Cu and Ni island films in visible and near-IR regions. In addition, the magneto-optic effect has been observed in the Cu-Ni CMI films. It is also shown that the Faraday rotation in the Cu-Ni CMI films can easily be controlled by selection of the mixture ratio of Cu and Ni.

Retrovirus in salivary glands from patients with Sjögren's syndrome.

AIMS: To investigate the possibility of an immune response to retroviral antigens or of detecting retrovirus in Sjögren's syndrome. METHODS: Retroviruses were sought in labial salivary glands and peripheral blood mononuclear cells from patients with Sjögren's syndrome by immunoblotting assay, immunohistochemical assay, polymerase chain reaction (PCR), reverse transcriptase (RT) activity assay, and transmission electron microscopy. RESULTS: Sera from five of 15 patients with Sjögren's syndrome (33%) reacted against p24 group specific antigen (gag) of human immunodeficiency virus (HIV). Labial salivary gland biopsy specimens from seven of the 15 patients with Sjögren's syndrome (47%) contained an epithelial cytoplasmic protein reactive with a monoclonal antibody to p24 of HIV. PCR was performed to detect HIV and human T lymphotropic virus type I (HTLV-I) genes from salivary gland tissues and peripheral blood mononuclear cells from patients with Sjögren's syndrome. Mn2+ dependent, Mg2+ independent RT activity was detected in the salivary gland tissues in three of 10 patients. A-type-like retroviral particles were observed in epithelial cells of salivary glands by transmission electron microscopy. Target genes for HIV and HTLV-I were not found in any of the salivary gland tissues or peripheral blood mononuclear cells from Sjögren's syndrome patients. CONCLUSIONS: The data suggest the presence of an unknown retrovirus similar to HIV in the salivary gland which might be involved in the pathogenesis of a subpopulation in Sjögren's syndrome.

Epstein-Barr virus (EBV) hypersensitivity of peripheral B lymphocytes in patients with EBV genome-positive Burkitt's lymphoma.

Peripheral blood mononuclear cells (PBMC) from four Japanese patients with Epstein-Barr virus (EBV) genome-positive Burkitt's lymphoma (BL) during remission were exposed to the B95-8 strain of EBV. Maximum concentrations of the EBV-determined nuclear antigen (EBNA) before cellular DNA synthesis were similar to those of healthy counterparts. Subsequently, EBV-immortalised cell lines were established. These immortalised lymphoblastoid cells were treated with 12-O-tetradecanoylphorbol-13-acetate (TPA) and superinfected with the P3HR-1 strain of EBV. EBV early antigens (EA) and viral capsid antigen (VCA) were expressed in approximately 3-10 fold higher concentrations by these lymphoblastoid cells than by those from patients with other types of malignant neoplasia including EBV genome-negative BL and from healthy counterparts. Moderate to extremely high IgG antibody titres to EBV VCA as well as IgG antibodies to EA were demonstrated in these patients during the study. These results suggest that defective underlying cellular mechanisms for regulating the replication of EBV may be present in patients with EBV genome-positive BL.

Decreased cytoplasmic immunoglobulin A production during Epstein-Barr virus immortalization on lymphocytes from patients with ataxia-telangiectasia.

Previously, we have reported significantly lower immunoglobulin (Ig) A production in supernatants of cultured lymphoblastoid cells using enzyme-linked immunosorbent assay from patients with ataxia-telangiectasia (AT) when compared to that of age- and sex-matched healthy individuals. Here, we further assess the degree of cytoplasmic Ig production in these cells and also analyze it during the early phase of Epstein-Barr virus immortalization. All classes of cytoplasmic IgM, IgG, and IgA productions were demonstrated in cells from healthy controls. In contrast, cells from patients with AT showed only cytoplasmic IgM and IgG with low or nondetectable levels of IgA during and after the immortalizing process. These results suggest B lymphocytes bearing IgA are functionally immature and/or defective in patients with AT.

African Burkitt's lymphoma: a plant, Euphorbia tirucalli, reduces Epstein-Barr virus-specific cellular immunity.

Endemicity of Burkitt's lymphoma (BL) coincides with profusion of a plant Euphorbia tirucalli in tropical Africa. E. tirucalli contains 4-deoxyphorbol ester that enhances Epstein-Barr virus (EBV) infection of B lymphocytes. In this study, we found that 4-deoxyphorbol ester reduced EBV-specific cytotoxic T-cell function. Furthermore, the B lymphocytes dually exposed to EBV and 4-deoxyphorbol ester were resistant to EBV-specific T cell cytotoxicity, through down-regulation of latent membrane protein 1 (LMP1), the major target to EBV-specific cytotoxic T-cells. These immunologic findings strengthen the notion that E. tirucalli may be an important environmental risk factor for the genesis of African BL.

High Epstein-Barr virus (EBV) susceptibility of both lymphoblastoid and lymphoma cell lines derived from a Japanese patient with EBV genome-positive Burkitt's lymphoma.

A Lymphoma cell line from the tumor tissue was established spontaneously from a Japanese patient with Epstein-Barr virus (EBV) genome-positive Burkitt's lymphoma (BL). Additionally lymphoblastoid cell lines from peripheral blood of this patient were established either spontaneously or by in vitro infection with B95-8 EBV. Lymphoma cells showed monoclonal surface immunoglobulins (kappa and gamma) with specific chromosomal translocations, t (8; 14). In contrast, lymphoblastoid cells expressed polyclonal surface immunoglobulins without specific chromosomal abnormalities. Lymphoma cells made colonies in soft agarose approximately 10 times more than those of the lymphoblastoid cells. When each cell line was cultured at lower temperature of 33 degrees C, treated with 12-O-tetradecanoyl- phorbol-13-acetate (TPA), and superinfected with P3HR-1 EBV, all cell lines expressed 5 to 10 times higher levels of EBV early antigens (EA) and viral capsid antigen (VCA) than lymphoblastoid cell lines from healthy controls. Furthermore, lymphoblastoid cell lines obtained from peripheral blood of this patient during the period of remission also exhibited high EA and VCA inducibility and superinfectibility. These findings suggested that the lymphoid cells in this patient were genetically highly susceptible to EBV infection, and this evidence possibly linked to the lymphomagenesis of EBV genome-positive BL.

Establishment of HIV-1-producing cells from peripheral mononuclear cells cultured with normal human serum.

Normal human serum (NHS) contributed to the establishment of cells producing HIV-1 under the conditions of coculture of peripheral mononuclear cells (PMC) from HIV-1 seropositive patients and of PHA-prestimulated or -non-stimulated PMC from seronegative healthy donors. No addition of IL-2 and Polybrene was necessary. Since, in the case 90101, the mitochondrial displacement-loop DNA showed identical sequences in the established cells and the HIV-1 seropositive patient's cells, it can be asserted that the HIV-1-producing cells originated from the patient. These cells are still releasing HIV-1-virion more than one year after their establishment.

Epstein-barr-virus hypersensitivity of lymphocytes from patients with ataxia-telangiectasia.

Ataxia telangiectasia (AT), an autosomal recessive disorder with a high incidence of lymphoreticular malignancies including Epstein-Barr virus (EBV)-induced lymphoproliferative disorders (LPD), was investigated to assess the susceptibility to EBV infection and oncogenesis. When the patients' lymphocytes were infected with B95-8 EBV, there was a tendency toward an enhanced growth in semisolid agar, as compared with the healthy donor counterparts. Among the preparations tested, from 14 patients, 2 cell lines showed extremely high colony forming efficiency. The lymphocytes from patients with AT did not contain a large number of EBV target cells, as determined by the maximum frequency of EBV-determined nuclear antigen (EBNA) induction prior to cellular DNA synthesis. Fourteen different lymphoblastoid cell lines derived from the 14 patients with AT were then examined for their EBV inducibility and superinfectibility. By treatment with 12-O-tetradecanoyl-phorbol-13-acetate TPA) and culturing at a lower temperature of 33-degrees-C, early antigen (EA) induction occurred approximately 6-fold and 5-fold higher, respectively, as compared with the lymphoblastoid cell lines derived from healthy controls. Viral capsid antigen (VCA) was also induced significantly by TPA or culturing at lower temperature in the lines from patients with AT, but only slightly in the control counterparts. When the lymphoblastoid cells from patients with AT were exposed to P3HR-1 EBV, EA and VCA syntheses were approximately 6- and 12-fold higher, respectively, than those in the cells derived from the healthy controls. This evidence suggested B lymphocytes of patients with AT were highly susceptible to EBV infection and possibly linked to the development of EBV-induced LPD.

High irradiation sensitivity of epstein-barr-virus (ebv) in lymphoblastoid-cells from patients with ataxia-telangiectasia and ebv genome-positive burkitts-lymphoma.

Epstein-Barr virus (EBV)-immortalized lymphoblastoid cell lines derived from the peripheral blood of patients with ataxia telangiectasia (AT) and EBV genome-positive Burkitt's lymphoma (BL) were tested for expression of EBV-related lytic antigens by means of irradiation. We used 1 Gy in each experiment, according to the results of the P3HR-1 (derived from African BL) cell line. Significantly higher expression of early antigens (EA) and viral capsid antigen (VCA) was demonstrated in lymphoblastoid cell lines derived from both patients with AT and those with EBV genome-positive BL, as compared to those derived from healthy individuals. These results suggested that defective regulating mechanisms on B lymphocytes, responsible for EBV infection, may underlie for the pathogenesis of development of lymphoproliferative diseases both in patients with AT and EBV genome-positive BL.