Characterization and comparison of two newly established Epstein-Barr virus (EBV)-negative and EBV-positive Burkitt's lymphoma cell lines. EBV-negative cell line with a low level of expression of ICAM-1 molecule and EBV-positive cell line with a high level of expression of ICAM-1 molecule.

Two human Burkitt's lymphoma cell lines (HBL-4 and HBL-5) were established individually from two patients with small noncleaved cell lymphoma (Burkitt's type). The HBL-4 cell line is Epstein-Barr virus (EBV)-negative, and the HBL-5 cell line is EBV-positive. Cytogenetically, both cell lines had the same chromosomal translocation, t(8;14)(q24;q32) as those observed in the primary malignant cells from individual patients. Morphologic, immunophenotypic, cytogenetic, and molecular studies confirmed that both cell lines were derived from the primary lymphoma cells in vivo. HBL-4 cells lacked CD23(H107), CD11a(LFA-1), and latent membrane protein (LMP) but expressed CD54(ICAM-1) at low levels, whereas HBL-5 cells showed the high level of expression of CD54 and faint expression of LMP but lacked CD11a. In addition, the EBV-positive lymphoblastoid cell line (LCL) expressed CD11a, CD23, CD54, and LMP at high levels. Therefore, an HBL-5 phenotype with expression of CD54 and LMP tends toward an LCL phenotype, and the augmentation of CD54 on the HBL-5 cells in comparison with primary lymphoma cells is likely to be upregulated by LMP, probably resulting from the EBV infection. There was little difference in the BrdUrd uptake in vivo and in vitro, doubling time, tumorigenicity, and dynamics of tumor growth in athymic nude mice between both cell lines. These findings indicate that the potentiality of cell growth and tumorigenicity of these two cell lines are unlikely to be related with EBV.     

A case of unruptured anterior temporal artery aneurysm showing pupil-sparing oculomotor palsy

The authors report a case of unruptured anterior temporal artery aneurysm showing pupil-sparing oculomotor palsy. The patient was a 55-year-old male with the complaints of left blepharoptosis and diplopia. He had a history of tuberculous meningitis 9 years previously, and since then he suffered from paraparesis of lower extremities and bladder and bowel disturbance. Neurological examination on admission revealed left blepharoptosis and disturbance of medial and vertical movement of the left eye ball, but the pupils were isocoric with normal light reaction. Carotid angiography demonstrated that the left internal carotid artery was tortuous toward the medial side in the C2 portion, and the saccular aneurysm was present in the anterior temporal artery 3 mm distal from the middle cerebral artery. At operation it was revealed that the aneurysm which had a diameter of 17 mm located in the proximal portion of the anterior temporal artery, and compressed the dorsal aspect of the oculomotor nerve. A month after operation the left oculomotor palsy disappeared. There had been no report of the case of unruptured middle cerebral artery aneurysm causing oculomotor palsy. Pupil-sparing oculomotor palsy is characteristic of diabetic oculomotor palsy, and rare cases showing pupil-sparing oculomotor palsy are caused almost by aneurysms of the internal carotid-posterior communicating artery junction. Anatomical study revealed that the parasympathetic fibers which go to the pupil constrictor muscle were in the dorsomedial aspect of the oculomotor nerve in the subarachnoid portion.(ABSTRACT TRUNCATED AT 250 WORDS)     

Suplatast tosilate inhibits thymus- and activation-regulated chemokine production by antigen-specific human Th2 cells

BACKGROUND: Suplatast tosilate is an anti-allergic agent that suppresses cytokine production by human Th2 cells. OBJECTIVE: We investigated the effects of suplatast tosilate on the production of thymus- and activation-regulated chemokine (TARC) by T cells from allergic patients with asthma. METHODS: Purified protein derivative (PPD)-specific Th1 cell lines and Dermatophagoides farinae (Der f)-specific Th2 cell lines were established from nine patients with house dust mite-allergic asthma. The effects of suplatast tosilate on mRNA expression of TARC and protein production of TARC from antigen-specific Th1 or Th2 cell lines were investigated after stimulation with relevant antigens or phytohemagglutinin (PHA). In addition, the effects of IL-4, IL-10, and IFN-gamma on TARC production by Der f-specific Th2 cell lines in the presence or absence of suplatast tosilate were studied. RESULTS: Although PPD-specific Th1 cell lines did not produce TARC after stimulation with PPD antigen or PHA, stimulation of Der f-specific Th2 cell lines with Der f antigen or PHA increased production of TARC. Suplatast tosilate significantly and dose-dependently inhibited production of TARC by Der f-specific Th2 cell lines stimulated with either Der f antigen (76.5% inhibition at 100 microg/mL, P < 0.01) or PHA (81.9% inhibition at 100 microg/mL, P < 0.01). TARC production by Der f-specific Th2 cell lines was significantly increased only by activation with IL-4 but not with IL-10 or IFN-gamma; this increase in TARC production was significantly inhibited by suplatast tosilate (97.5% inhibition at 100 microg/mL, P < 0.01). CONCLUSION: Suplatast tosilate inhibits TARC production by human Th2 cells. Therefore, this agent inhibits both Th2 cytokine and Th2 chemokine and may be a useful anti-allergic agent.     

Three-dimensional two-layer collagen matrix gel culture model for evaluating complex biological functions of monocyte-derived dendritic cells

Dendritic cell-like cells (Mo-DCs) generated from peripheral blood monocytes with interleukin-4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF) have been used as tools to treat cancer patients (DC-vaccines). Because Mo-DCs have multiple antigen presentation-related functions, including phagocytosis, migration, cytokine production, and T cell stimulation, establishment of a method for simultaneously evaluating the various functions of Mo-DCs is important. We developed a new in vitro three-dimensional two-layer collagen matrix culture model that consists of a collagen gel containing Mo-DCs as the lower layer and a collagen gel containing necrotic GCTM-1 tumor cells and/or T cells as the upper layer. We used this system to observe simultaneously multiple functions of Mo-DCs by phase-contrast or fluorescence microscopy and to assess IL-12 secretion during more than 2 weeks of culture. We also observed interactions between Mo-DCs and necrotic GCTM-1 or T cells on an individual cell basis by time-lapse videomicroscopy. In addition, we collected Mo-DCs from the collagen gels by collagenase treatment and analyzed the expression of antigen presentation-related molecules such as HLA-DR, CD80, CD83, and CD86 on Mo-DCs. This model may be a useful tool for evaluation of the various functions of Mo-DCs used as DC vaccines and for studies of the complex behaviors of Mo-DCs in vivo.     

Clinical studies on cefminox in pediatric field

Clinical studies on cefminox (CMNX, MT-141) were conducted and the following results were obtained. Twelve cases of bacterial infections were treated with CMNX with a satisfactory result of "excellent" in 9 and "good" in 3. Antibacterial activity was examined in 7 cases. Pathogenic organisms which were S. aureus, beta-Streptococcus, H. influenzae, S. pneumoniae and P. morganii were eradicated in all the cases. Mean maximum serum concentrations of CMNX after intravenous injection of 10, 20 and 40 mg/kg were 54.5 mcg/ml, 102.3 mcg/ml and 202.4 mcg/ml, respectively which were obtained 15 minutes after each injection. Mean half-lives of each dose group were 1.60, 1.13 and 1.51 hours, respectively. Mean urinary excretion rates of CMNX at 6 hours after intravenous injection in 10, 20, 30 and 40 mg/kg groups were 73.5%, 80.9%, 92.6% and 66.5%, respectively. Side effects were not observed clinically, but anemia in 1 case and eosinophilia in 2 cases were noted in laboratory examination.     

Carpal Tunnel Syndrome Due to Iatrogenic Amyloidosis After Domino Liver Transplantation From Hereditary Transthyretin Amyloidosis: A Case Report

BackgroundCarpal tunnel syndrome is the most common compression syndrome of the peripheral nerve. Transthyretin amyloidosis and dialysis-related β2-microglobulin amyloidosis are known causes of carpal tunnel syndrome.Case ReportA Japanese woman showed carpal tunnel syndrome 16 years after a domino liver transplantation (DLT) from the donor with hereditary transthyretin amyloidosis. DLT indication was congenital extrahepatic portosystemic shunt, and the patient had been put on maintenance hemodialysis because of chronic kidney disease 6 years before DLT. Moreover, the amyloid precursor protein of the patient was histologically confirmed not to be β2-microglobulin, but transthyretin.ConclusionsThe existence of amyloid was speculated when the patient who underwent DLT from hereditary transthyretin amyloidosis showed carpal tunnel syndrome. Additionally, elucidating the amyloid precursor protein when the patient has another cause of amyloidosis is necessary.     

Strain differences in age-associated change in testosterone 6β-hydroxylation in Wistar and Dark Agouti rats

This study examines strain differences in testosterone (T)-hydroxylations between Wistar and Dark Agouti (DA) rats of both genders. The DA rat, an animal model, is a poor metabolizer of such drugs as debrisoquine, which are metabolized by cytochrome P450 (CYP) 2D. T-16α-, 2α-hydroxylations, which are linked to CYP2C11, were catalyzed at similar rates by the microsomes of both strains. In contrast, the liver microsomes from mature male DA rats catalyzed T-6β-hydroxylation, the CYP3A mediated activity, at higher rates (～ 2-fold) than Wistar rat liver microsomes did. There was no difference between immature male DA and Wistar rats for T-6β-hydroxylation, indicating that the activity in male DA rat increases with maturation. Polyclonal antibodies raised against rat liver microsomal CYP3A2 and a CYP3A inhibitor, troleandomycin (TAO), effectively inhibited T-6β-hydroxylation by liver microsomes from both strains of rats. The level of T-6β- hydroxylation activity correlated well with the amount of CYP3A protein in the microsomes in mature as well as in immature male and female Wistar and DA rats. Northern blot analysis repeatedly indicated that the cellular contents of CYP3A2 mRNA are slightly (～ 20%) higher in the liver of mature DA rats than in that of mature Wistar rats. These results indicate that the increased levels of CYP3A are responsible for the increased T-6β-hydroxylation activity and protein in DA rat.     

Tolerance of Vascularized Islet‐Kidney Transplants in Rhesus Monkeys

We previously reported that transplantation (Tx) of prevascularized donor islets as composite islet‐kidneys (IK) reversed diabetic hyperglycemia in both miniature swine and baboons. In order to enhance this strategy's potential clinical applicability, we have now combined this approach with hematopoietic stem cell (HSC) Tx in an attempt to induce tolerance in nonhuman primates. IKs were prepared by isolating islets from 70% partial pancreatectomies and injecting them beneath the autologous renal capsule of five rhesus monkey donors at least 3 months before allogeneic IK Tx. HSC Tx was performed after mobilization and leukapheresis of the donors and conditioning of the recipients with total body irradiation, T cell depletion, and cyclosporine. One IK was harvested for histologic analysis and four were transplanted into diabetic recipients. IK Tx was performed either 20–22 (n = 3) or 208 (n = 1) days after HSC Tx. All animals accepted IKs without rejection. All recipients required >20 U/day insulin before IK Tx to maintain <200 mg/dL, whereas after IK Tx, three animals required minimal doses of insulin (1–3 U/day) and one animal was insulin free. These results constitute a proof‐of‐principle that this IK tolerance strategy may provide a cure for both end‐stage renal disease and diabetes without the need for immunosuppression.