Membrane-bound form of monocyte chemoattractant protein-1 enhances antitumor effects of suicide gene therapy in a model of hepatocellular carcinoma.

Suicide gene therapy using the herpes simplex virus thymidine kinase/ganciclovir (HSV-tk/GCV) system combined with monocyte chemoattractant protein-1 (MCP-1) provides significant antitumor efficacy. The current study was designed to evaluate the antitumor immunity of a newly developed membrane-bound form of MCP-1 (mMCP-1) in an immunocompetent mouse model of hepatocellular carcinoma (HCC). A recombinant adenovirus vector (rAd) harboring the human MCP-1 gene and the membrane-spanning domain of the CX3CL1 gene was used. Large amounts of MCP-1 protein were expressed and accumulated on the tumor cell surface. The growth of subcutaneous tumors was markedly suppressed when tumors were treated with mMCP-1, as compared with soluble MCP-1, in combination with the HSV-tk/GCV system (P<0.01). The numbers of Mac-1-, CD4- and CD8a-positive cells were significantly higher in tumor tissues (P<0.05), and tumor necrosis factor (TNF) mRNA expression levels with mMCP-1 were almost five-fold higher than those with soluble MCP-1. These results indicate that the delivery of the mMCP-1 gene greatly enhanced antitumor effects following the apoptotic stimuli by promoting the recruitment and activation of macrophages and T lymphocytes, suggesting a novel strategy of immune-based gene therapy in the treatment of patients with HCC.

Successful embolisation of intrahepatic portosystemic venous shunt using coils and n-butyl cyanoacrylate through two approach routes.

Interventional radiological treatment by transcatheter embolisation has been used to treat patients with symptomatic intrahepatic portosystemic venous shunt (IPSVS). We present a case of symptomatic IPSVS treated by both retrograde and antegrade transcatheter embolisation using coils and n-butyl cyanoacrylate.

Facial emphysema caused by cheek bite.

Biting of the buccal mucosa is very frequent injury, whereas facial emphysema caused by cheek bite is rare. We report a case of facial emphysema causing puffing of the cheek through a self-inflicted bite of the buccal mucosa.

A polymorphism of DRD2 gene and brain atrophy in methamphetamine psychosis.

Our group, Ujike et al., recently reported that the A1 allele of TaqI A polymorphism of the dopamine receptor D2 (DRD2) gene, associated with transient psychosis, significantly differs from that of patients with prolonged psychosis in methamphetamine psychosis. Therefore, we examined the association between the TaqI A polymorphism of the DRD2 gene and the brain MRI view for patients with methamphetamine psychosis. The subjects underwent brain MRI scans using the FLAIR method. Genotyping was performed by PCR-RFLP methods using genomic DNA extracted from peripheral blood by the phenol method. Ten subjects had the A1/A2 genotype, eleven subjects had the A2/A2 genotype, and no subject had the A1/A1 genotype. The domain size, including the thalamus and basal ganglia that were inside each side of the putamens, did not differ between the three groups (the A1/A2-group, the A2/A2-group, and the young healthy person group). In the comparison based on this domain, the temporal lobe tended to narrow in the A2/A2-group compared to the A1/A2-group (P = .06). The other domain (cerebrum, corpus callosum, etc.) showed no difference between the A1/A2-group and the A2/A2-group. It is suggested that in methamphetamine psychosis the TaqI A polymorphism not only regulates prolongation of psychosis symptoms but also influences the form of the temporal lobe.

Possible importance of immunoglobulin E in foetal loss by mothers with anti-SSA antibody.

OBJECTIVE: The incidence of foetal loss and/or adverse foetal outcomes, including congenital heart block (CHB), has been investigated in mothers with anti-SSA antibody detected by immunodiffusion or counter-immunoelectrophoresis methods. We investigated the relationship between several serum parameters (such as autoantibodies and immunoglobulins) and foetal loss in patients with anti-SSA antibody, measured by enzyme-linked immunosorbent assay (ELISA). MATERIAL AND METHODS: Thirty-seven women who showed positivity for anti-SSA antibody and had a history of pregnancy were included in this study. Immunoglobulins and several autoantibodies were assayed by routine laboratory methods at our hospital. RESULTS: Our data indicated that immunoglobulin E (IgE) levels were significantly higher in the anti-SSA antibody positive women with foetal loss than in those without, and that a strong positive correlation between IgE and anti-SSA antibody levels was observed in the former group, but not in the latter. CONCLUSION: The serum IgE level seems to be an important factor in the occurrence of foetal loss in mothers with anti-SSA antibody detected by ELISA.

Systemic lupus erythematosus-like autoimmune abnormalities induced by bacterial infection.

Recent evidence has revealed that bacterial DNA can promote several of the autoimmune abnormalities observed in systemic lupus erythematosus (SLE), and a possible pathogenic role in the induction of SLE has been highlighted. We have recently encountered patients in whom bacterial infection (septicemia) triggered the production of several autoantibodies. This seems to be interesting with respect to the consideration of the relationship between SLE and bacterial infection.

Allergic diseases in systemic lupus erythematosus: prevalence and immunological considerations.

We attempted to obtain a deeper understanding of the relationship between systemic lupus erythematosus (SLE) and allergic diseases through comparative studies. Accordingly, we reviewed the association of both disorders and compared their immunological features based on the literature and our own findings. Recent studies (including ours) have indicated that the risk of IgE-mediated and/or associated allergic diseases is not markedly increased in SLE patients despite their more allergic family history when compared with controls, in contrast with earlier studies. This may be related to a change of the environmental factors contributing to allergy. In addition, assessment of the immunological similarities and differences between SLE and various allergic diseases seems to be useful for understanding the relationship between them.

Determination of residual double bonds in resin-dentin interface by Raman spectroscopy.

OBJECTIVES: The quality of the hybrid layer is believed to be more important than the thickness of this layer. The purpose of this study was to investigate a method to analyze the percentage of adhesive resin residual double bonds in the dentin-resin interface using laser Raman spectroscopy. METHODS: Bovine dentin was treated with dentin adhesives and resin composite was bonded according to the manufacturers' instructions. The specimens were sectioned parallel to dentinal tubules and the surfaces were then polished to 1 microm diamond pastes. Raman spectra were recorded along a line perpendicular to the dentin-resin interface in steps of 0.2 microm. The measurement of residual C=C bond was made on a relative basis by comparing the C=C unpolymerized methacrylate stretching vibration (1638 cm(-1)) against the C=O stretching mode of the ester group (1719 cm(-1)). The percentage of residual double bonds including pendant and monomeric double bonds was calculated by comparing the obtained ratio with that of uncured adhesive resin. RESULTS: The amount of residual double bonds in the hybrid layer varied from 10 to 25% compared to the uncured adhesives, a relatively higher percentage was detected for Fluoro Bond (12.3-23.6%) and Single Bond (9.5-21.8%), and lower for Mac Bond II (10.6-18.0%) and Mega Bond (10.7-16.3%). No relationship was seen between the percentage of remaining double bonds and the location within the resin-dentin interface. SIGNIFICANCE: Laser Raman microscopy used was a useful tool for measuring the residual double bonds in the dentin-resin interface.

Allergic disorders in systemic lupus erythematosus: prevalence and family history.

The relationship between allergic disorders (such as atopic dermatitis, asthma, allergic rhinitis and allergic conjunctivitis) and systemic lupus erythematosus (SLE) is still unclear and controversial. We investigated the prevalence of these allergic disorders in SLE patients and their families. A questionnaire about the history of allergy was completed by 52 SLE patients and by 52 matched (including race, age, sex and region) non-SLE controls. Our results indicated that there was a significantly lower incidence of these allergic diseases in SLE patients, especially those who had an allergic family histories, when compared with the controls. These findings may be related to the immunological similarities and differences between SLE and various allergic diseases.

Cytomegalovirus infection in patients with systemic lupus erythematosus.

Cytomegalovirus (CMV) infection is known to induce several autoimmune abnormalities in mice that resemble those found in systemic lupus erythematosus (SLE). In addition, a potential role for CMV in the development and/or progression of SLE has been suggested. In order to further clarify this issue, we reviewed the relationship between SLE and CMV infection on the basis of the clinical and immunological features of cases reported in the literature and our own patients.

Lessons from similarities between SLE and HIV infection.

OBJECTIVE: We attempted to obtain deeper understanding of systemic lupus erythematosus (SLE) and human immunodeficiency virus (HIV) infection through comparative studies between both diseases. METHOD: For this purpose, we reviewed and discussed lessons from similarities in both diseases based on our own and reported findings in literatures. RESULT: Such comparative studies may contribute to the progress in understanding the clinical or pathogenetic features of these diseases. CONCLUSION: Further studies into the relationship between SLE and HIV infection may bring to light important clues to assist in the development of better treatments for each disease.

Cyclosporin A therapy for interstitial pneumonitis associated with rheumatic disease.

Abstract To determine the efficacy of cyclosporin A (CysA) for the treatment of steroid-resistant interstitial pneumonitis (IP), we enrolled 25 patients with various rheumatic diseases and steroid-resistant IP in a pilot study [4 patients with rheumatoid arthritis (RA), 2 with systemic lupus erythematosus (SLE), 11 with polymyositis/dermatomyositis (PM/DM), 4 with systemic sclerosis (SSc), 1 with mixed connective tissue disease (MCTD), 3 with Sjögren syndrome (SS)]. Twelve patients (48%) showed a persistent response to CysA therapy, and 7 of them had PM/DM, including so-called amyopathic DM. Patients with a persistent response had moderately elevated lactate dehydroxygenase (LDH) levels, whereas patients who died had much higher LDH levels and hypoxia. Even patients with low blood levels of CysA achieved a persistent response. In responding patients, the symptoms, chest X-ray findings, arterial oxygen tension, and LDH level all improved after less than 4 weeks. In conclusion, CysA seem to be useful for treating patients with steroid-resistant IP, whose duration is short and severity is mild.

Discrete roles of cytokines, TNF-alpha, IL-1, IL-6 in tumor promotion and cell transformation.

Based on our previous results, which pointed to tumor necrosis factor-alpha (TNF-alpha) as the essential cytokine in tumor promotion in mouse skin, we present here three principal findings related to the specific roles of TNF-alpha, interleukin-1 (IL-1) and IL-6 in tumor promotion (using TNF-alpha- and IL-6-deficient mice) and in BALB/3T3 cell transformation: i) The previously reported residual tumor promotion by 12-O-tetradecanoylphorbol-13-acetate (TPA) in TNF-/- mice was confirmed by experiments with TNF+/+ and TNF-/- 129/Svj mice of the same strain, using two-stage carcinogenesis experiments. TPA produced tumors in 100% of TNF+/+ and 78% of TNF-/- mice at 20 weeks, and the average number of tumors per mouse was 11.1 in the former group and 2.1 in the latter. Judging from the expression of various inflammatory cytokine genes in TNF+/+ and TNF-/- mice, the residual tumor promoting activity of TPA in TNF-/- mice may be dependent on expression of IL-1alpha and IL-1beta genes. ii) Tumor promotion by TPA and okadaic acid in IL-6+/+ and IL-6-/- C57/BL6 mice was studied, with TPA producing tumors in 57.1% of IL-6+/+ and 40.0% of IL-6-/- mice at 20 weeks, and okadaic acid in 40.0% of IL-6+/+ and 53.3% of IL-6-/- mice. Thus, there was no significant difference between TPA or okadaic acid tumor promotion in either group. In addition, expression of IL-6 gene in skin of both types of mice suggested that IL-6 is not the essential cytokine in tumor promotion, since it can be replaced by other cytokines. iii) In transformed clones of BALB/3T3 cells induced by TNF-alpha alone, IL-1alpha gene expression was induced after transformation by TNF-alpha had occurred, which did not occur in parental cells. Expression patterns of TNF-alpha, IL-1beta, IL-6 and IL-10, along with TGF-beta, were similar in both parental and transformed cells. Considering all these results, we conclude that various cytokines have discrete roles in tumor promotion and cell transformation.

Reversal effects of antifungal drugs on multidrug resistance in MDR1-overexpressing HeLa cells.

In this study, the antiproliferative effects of vinblastine (VLB), paclitaxel (TXL), doxorubicin (DXR), daunorubicin (DNR) and 5-fluorouracil (5-FU) were assessed in the human cervical carcinoma cell line HeLa-Ohio (HeLa) and Hvr100-6 cells, established by growing the parental HeLa cells in the presence of progressively greater concentrations of VLB in the culture medium. Flow cytometric analysis indicated the induction of MDR1 (P-glycoprotein) in Hvr100-6 cells with no alterations in levels of multidrug resistance-associated protein (MRP). Resistance to VLB, TXL, DXR and DNR was found in Hvr100-6 cells with relative resistances of ca. 300, 4000, 50 and 200, respectively, whereas no resistance was found to 5-FU. The reversal effects of antifungal drugs, fluconazole, itraconazole, ketoconazole, miconazole and amphotericin B on multidrug resistance were also assessed using Hvr100-6 cells. Itraconazole was found to have potent reversal effect on the resistance to VLB and TXL, but the others had no such effect. This reversal effect of itraconazole was concentration-dependent, with dose modifying factors of 3.2, 10.1 and 435.7 at 0.1, 0.25 and 0.5 microM of itraconazole, respectively. In addition, this reversal effect of itraconazole was explained by the inhibition of accumulation of the anticancer drugs.

Requirement of Ras for the activation of mitogen-activated protein kinase by calcium influx, cAMP, and neurotrophin in hippocampal neurons.

Mitogen-activated protein (MAP) kinase plays important roles in the establishment of long-term potentiation both in vitro and in living animals. MAP kinase is activated in response to a broad range of stimuli, including calcium influx through NMDA receptor and L-type calcium channel, cAMP, and neurotrophins. To investigate the role of Ras in the activation of MAP kinase and cAMP response element-binding protein (CREB) in hippocampal neurons, we inhibited Ras function by overexpressing a Ras GTPase-activating protein, Gap1(m), or dominant negative Ras by means of adenovirus vectors. Gap1(m) expression almost completely suppressed MAP kinase activation in response to NMDA, calcium ionophore, membrane depolarization, forskolin, and brain-derived neurotrophic factor (BDNF). Dominant negative Ras also showed similar effects. On the other hand, Rap1GAP did not significantly inhibit the forskolin-induced activation of MAP kinase. In contrast to MAP kinase activation, the inactivation of Ras activity did not inhibit significantly NMDA-induced CREB phosphorylation, whereas BDNF-induced CREB phosphorylation was inhibited almost completely. These results demonstrate that Ras transduces signals elicited by a broad range of stimuli to MAP kinase in hippocampal neurons and further suggest that CREB phosphorylation depends on multiple pathways.