High mortality rate of (NZW x BXSB)F1 mice induced by administration of lipopolysaccharide attributes to high production of tumour necrosis factor-alpha by increased numbers of dendritic cells.

(NZW x BXSB)F1 mice (W/BF1 mice) have been reported to be a type of autoimmune-prone mice, showing symptoms of proteinuria, anti-DNA antibodies and anti-platelet antibodies. In this paper, we report that W/BF1 mice show hyperproduction of tumour necrosis factor (TNF)-alpha, responding to lipopolysaccharide (LPS) in comparison with normal mice, resulting in induction of death. In normal mice, monocytes/macrophages (Mo/MØ) are the main producer of TNF-alpha, while both Mo/MØ and dendritic cells (DCs) produce TNF-alpha in W/BF1 mice. Because the number of DCs is higher in W/BF1 mice, the main producers of TNF-alpha in W/BF1 mice are thought to be DCs. Moreover, administration of anti-TNF-alpha antibodies rescued the W/BF1 mice from death induced by LPS, suggesting that TNF-alpha is crucial for the effect of LPS. Although there is no significant difference in the expression of Toll-like receptor-4 (TLR-4) on DCs between B6 and W/BF1 mice, nuclear factor kappa b activity of DCs from W/BF1 mice is augmented under stimulation of LPS in comparison with that of normal mice. These results suggest that the signal transduction from TLR-4 is augmented in W/BF1 mice in comparison with normal mice, resulting in the hyperproduction of TNF-alpha and reduced survival rate. The results also suggest that not only the quantity of endotoxin, but also the host conditions, the facility to translate signal from TLR, and so on, could reflect the degree of bacterial infections and prognosis.

Intra-bone marrow injection of dendritic cells for treatment of malignant tumor in mice.

It has recently been reported that antigen presentation from dendritic cells (DCs) to T cells occurs in the bone marrow, and that not only tumor antigen-pulsed DCs but also unpulsed DCs have some anti-tumor effects, resulting from the induction of anti-tumor immunity. In this paper, we examined whether dendritic cells induced from bone marrow cells (BMCs) have the capacity to suppress tumor growth and, if so, which route (intravenous, subcutaneous, or intra-bone marrow injection) is best. BALB/c mice that had been subcutaneously inoculated with Meth A (a murine fibrosarcoma cell line) were injected with BMC-derived DCs via the above three routes. We also examined the tumor suppressive effects of DCs from tumor-bearing mice. Although IBM injection showed similar effects to subcutaneous injection on the suppression of tumor growth, intravenous injection was less effective. It seems likely that the IBM injection of DCs activates tumor-specific T cells, resulting in the suppression of the tumor growth. DCs derived from tumor-bearing mice had some effects on the suppression of tumor growth but they were less effective than DCs from untreated mice.

Synergistic effects of injection of bone marrow cells into both portal vein and bone marrow on tolerance induction in transplantation of allogeneic pancreatic islets.

We have established a new method for allogeneic pancreatic islet (PI) transplantation: relatively low doses of irradiation followed by simultaneous transplantation of PIs and bone marrow cells (BMCs) via the portal vein (PV). In the present study, we have compared this method with intra-bone marrow (IBM)-bone marrow transplantation (BMT), and with a combination of both methods. Streptozotocin (STZ)-induced diabetic-recipient rats, Fischer 344 (F344, RT1A(l), RT1B(l)), were irradiated 1 day before transplantation. PIs of Brown Norway rats (BN, RT1A(n), RT1B(n)) were transplanted into the liver of the diabetic F344 rats via the PV. BMCs from BN rats were injected into the recipients' bone marrow (IBM), PV or intravenously (IV) or by a simultaneous combination of PV plus IBM (PV+IBM). We compared graft survival among the groups of '9 Gy+IBM'(10/10 accepted), '9 Gy+PV'(7/10 accepted), '9 Gy+IV'(0/7 accepted), '9 Gy+PV+IBM'(8/8 accepted), '8.5 Gy+IBM'(4/9 accepted), '8.5 Gy+PV'(0/7 accepted), '8.5 Gy+IV'(0/7 accepted), '8.5 Gy+PV+IBM'(9/12 accepted), '8 Gy+IBM'(2/10 accepted) and '8 Gy+PV+IBM'(2/8 accepted). As we reported previously, PV-BMT is more effective in inducing the acceptance of allogeneic PIs than IV-BMT. However, IBM-BMT requires less pretreatment than PV-BMT. (PV+IBM)-BMT was found to be the most effective in inducing the acceptance of allogeneic PIs. These results suggest that allogeneic PI-transplantation in conjunction with (PV+IBM)-BMT could become a viable strategy.

Relationship between the structure and cytotoxic activity of asterriquinone, an antitumor metabolite of Aspergillus terreus, and its alkyl ether derivatives.

Asterriquinone (ARQ) is an antitumor metabolite of Aspergillus terreus IFO 6123. In this study we synthesized several ARQ alkyl ethers and studied their cytotoxic activity against mouse leukemia P388 cells. The dissociation constant was also similar among the ARQ monoalkyl ethers. There was a good correlation between the hydrophobicity of ARQ monoalkyl ethers estimated by high performance liquid chromatography and the intracellular content accumulated for 60 min. ARQ monoalkyl ethers were cytotoxic, but ARQ dimethyl ether was not, as previously reported. The cytotoxicity of the ARQ monoalkyl ether derivatives was increased with extension of the alkyl chain length. The cytotoxicity was closely correlated with the intracellular content. The strongest ARQ derivative, ARQ monohexyl ether (ARQHex), formed more DNA-interstrand cross-links in the cells than ARQ. After treatment of P388 cells with ARQ and ARQHex for 6 h, a nucleosomal ladder pattern, as well as the appearance of degraded DNA, observed by flow cytometry, indicated that these compounds caused cellular apoptosis. Moreover, ARQ and ARQHex accumulated in the cells at the G1 phase of the cell cycle. These results indicated that ARQ monoalkyl ethers increased cytotoxicity, according to their membrane permeability based on the hydrophobicity, and they caused apoptotic cell death, as did ARQ.

Mechanism of the cytotoxicity of asterriquinone, a metabolite of Aspergillus terreus.

Asterriquinone (ARQ) is an antitumor metabolite of Aspergillus terreus IFO 6123. ARQ is a quinone compound and the mechanism of its antitumor action was compared with that of its analog ARQDMe which was dimethylated at the dihydroxy benzoquinone moiety, and adriamycin (ADR). The cytotoxicity of ARQ against P388 cells was less than that of ADR, and ARQDMe showed hardly any cytotoxicity. ARQ and ADR formed DNA-interstrand cross links (ISC) in P388 cells in a concentration while the ISC index by ARQDMe was very low even at 10 microM. P388 cells treated with ARQ and ADR for 18 hours showed, dose-dependently, nucleosomal ladder formation as well as the appearance of degraded DNA. Difference between the action of ARQ and ADR were observed by flow cytometric cell cycle analysis. ARQ caused the cells to accumulate at the G1 phase of the cell cycle, while use of ADR led to arrest in the G2/M phase. ARQDMe never caused apoptotic changes or cell cycle alterations. These results indicate that ARQ intercalates in the genomic DNA, causes G1 arrest, and leads to apoptotic cell death, this suggests that the dihydroxybenzoquinone moiety of the compound is essential to elicit these actions.

Four new metabolites of Aspergillus terreus.

Four new metabolites (1-4) were isolated from mycelium of Aspergillus terreus IFO 6123 producing asterriquinone (ARQ). The structures of 1 and 2 were shown to be 3,6-bis[1-(1,1-dimethyl-2-propenyl)-1H-indol-3-yl]-furo[3,2-b]furan- 2,5-dione (asterridinone) and 2,5-bis[1-(1,1-dimethyl-2-propenyl)-1H-indol-3- yl]-3-acetoxy-6-hydroxy-2,5-cyclohexadiene-1,4-dione (ARQ monoacetate), respectively, by the chemical and spectral data. Compounds 3 and 4 were identified with known asterriquinone isomers, 2-[1-(1,1-dimethyl-2-propenyl)-1H-indol-3-yl]-5-[2-(3-methyl-2-butenyl)- 1H-indol-3-yl]-3,6-dihydroxy-2,5-cyclohexadiene-1,4-dione (isoARQ) and 2,5-bis[2-(3-methyl-2-butenyl)-1H-indol-3-yl]-3,6-dihydroxy- 2,5-cyclohexadiene-1,4-dione (neoARQ), respectively.

Diagnosis of right ventricular involvement in chronic inferior myocardial infarction by means of body surface QRS changes.

ST segment elevation in right precordial leads is thought to be good predictor of right ventricular involvement in patients with acute inferior myocardial infarction. This view, however, is rapidly disappearing. Therefore, using QRS changes in body surface potential maps in the chronic phase, we have attempted to differentiate patients with or without right ventricular involvement. Thirty patients with chronic inferior myocardial infarction (2 or more months after onset) were studied, in whom 87 unipolar ECGs and right ventriculograms were recorded. The patients were then divided into three groups depending on the locations of their abnormal QRS potentials (-2SD area) exceeding the normal range (mean -2SD). In group A, the -2SD area was located predominantly on the right inferior chest, in group B on the left inferior chest, and in group N on both the right and left inferior chests equally. The results showed that group A had a lower right ventricular ejection fraction (RVEF) compared with group B (A, 40 +/- 7%; B, 53 +/- 10%; p less than .001), while there was no difference in left ventricular ejection fraction between the two groups (49 +/- 11% and 49 +/- 11%, respectively). Moreover, right ventricular asynergy occurred in 14 of the 18 patients (78%) of group A but in only one of the 10 patients (10%) of group B. Group N was presumed to be intermediate between groups A and B.(ABSTRACT TRUNCATED AT 250 WORDS)