Accumulation of cell cycle regulatory proteins, p21 and p27, induced after hyperthermia in human glioma cells.

It was investigated whether there was a relationship between p53 p21 and p27 induction pathways in the cellular response of glioma cells to hyperthermia. Two glioma cell lines were employed. A-172 cells had the wild-type of p53, and U251 cells had the mutant-type of p53. An adenovirus harbouring wild-type p53 was also used for the overexpression. The protein induction by hyperthermia was monitored by Western blot analysis. In U251 cells, the expression of wild-type p53 and hyperthermia had an additional cytotoxic effect, but did not affect A-172 cells. Significant p21 accumulation by hyperthermia was recognized in A-172 cells, and was also recognized in p53-transduced U251 cells. On the other hand, the accumulation of p27 by hyperthermia was not seen in A-172 or U251 cells, and the exogenous expression of p53 did not affect the accumulation of p27 by hyperthermia in U251 cells. These findings suggest that the p53-p21 pathway is involved in the signal transduction after hyperthermia, rather than the p27 pathway.

[Neurosurgical therapy for parkinson's disease].

Stereotactic surgery and neuronal transplantation are considered to be effective surgical therapies for advanced Parkinson's disease with wearing off phenomenon. As a stereotactic surgery, posteroventral pallidotomy and chronic pallidal or subthalamic stimulation with inhibitory parameters were performed. Flattening of the motor fluctuations were obtained with improving general motor symptoms especially at "off" time. Therapeutic l-dopa dose could not reduced following pallidotomy and pallidal stimulation, whereas subthalamic stimulation saved 30-50% of l-dopa dose. By the constant supply of dopamine, neuronal transplantation was effective on wearing off. Chromaffin cells of adrenal medulla and pretransected peripheral nerve were cografted into the bilateral caudate nuclei. After the transplantation significant reduction of % time off and l-dopa dose was observed.

[Combination effect of hyperthermia and MX2 on cultured glioma cell lines].

MX2, a new lipophilic morpholino anthracycline, has been reported to have chemotherapeutic effects superior to those of adriamycin against murine and human glioma cells in vitro and in vivo. To assess the combination effect of MX2 and hyperthermia in vitro, the thermo-chemosensitivities of cultured human (U251MG and KC) and rat (C6 and 9L) glioma cell lines were examined by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay. The number of viable cells in each cell line was markedly and dose-dependently decreased by MX2 treatment, but the sensitivity of U251MG to MX2 was less than that of the other cell lines. The survival of each cell line was decreased with the hyperthermic treatment at 43 degrees C for 60 minutes. Combined treatment with MX2 and hyperthermia had an additive effect on cultured glioma cells when MX2 was added to the medium at a dose below 50 ng/ml. However, combined treatment indicated neither an additive nor a synergistic effect when the dose of MX2 was above 50 ng/ml. We conclude that MX2 may be clinically useful against malignant gliomas when administered alone or in combination with hyperthermia.

Primary anorectal malignant melanoma: report of a case.

Primary anorectal malignant melanoma is a fairly uncommon but highly malignant disease. This disease is sometimes mistaken for such benign conditions as either a hemorrhoid or rectal polyp. We herein describe a case of early primary malignant melanoma of the anal canal. In this case, magnetic resonance (MR) imaging was found to be useful for diagnosing the melanotic melanoma. We especially emphasize the usefulness of a fat-saturation MR image in distinguishing melanotic melanoma from other rectal tumors.

[Primary malignant lymphoma in the central nervous system treated with high dose methotrexate (MTX)-CHOP (M-CHOP)].

From December 1995 to July 1997, six patients with primary malignant lymphoma in the central nervous system were treated with 2 to 5 cycles of the M-CHOP regimen (methotrexate 3 g/m2 on day 1, cyclophosphamide 750 mg/m2 on day 1, doxorubicin 40 mg/m2 on day 1, vincristine 1.4 mg/m2 on day 1, and predonisolone 60 mg on day 1 to 14: folic acid was given 3 hours after methotrexate at 10 mg/m2 every 3 hours for 9 doses intravenously). Five patients achieved complete remission (CR) and one experienced partial remission (PR). Posttherapeutic studies were performed in all patients with an average follow-up period of 20.1 months (range 8.1-26.8 months) after confirming the diagnosis. There was no evidence of recurrence of the tumors or growth of residual tumors in any of the patients in this period. The major toxic effect was myelosupression with leukopenia. Alopecia was observed in all patients. No treatment-related deaths were observed. The M-CHOP regimen seems to be a promising treatment for primary malignant lymphoma in the central nervous system.

Proliferative potential and apoptosis in rat glioma cell lines after hyperthermia.

The proliferative potential of cultured rat glioma cells (C6 and 9L) was evaluated after hyperthermia using immunohistochemical staining with bromodeoxyuridine (BrdU) and Ki-67 monoclonal antibodies. Apoptosis was assessed by in situ end-labeling of deoxyribonucleic acid breaks. Both BrdU and Ki-67 labeling indexes decreased with increasing hyperthermia time. The decrease of the Ki-67 labeling index was not as great as that of the BrdU labeling index. The number of apoptotic cells increased with time after hyperthermia. These results indicate that the antitumor effect of hyperthermia may reflect the induction of apoptosis in the cells within the cell cycle, and the resultant reduction of the proliferative potential of surviving cells, especially in the S phase.

The protective effect of dexamethasone against radiation damage induced by interstitial irradiation in normal monkey brain.

OBJECTIVE: The protective effect of dexamethasone against radiation damage is unclear. We examined the effect of early treatment of high-dose dexamethasone on iridium-192-induced damage to normal brain tissue. METHODS: Brain damage induced by interstitial irradiation with iridium-192 was evaluated with sequential magnetic resonance imaging and proton magnetic resonance spectroscopy in 11 adult monkeys, with or without short-term high-dose dexamethasone treatment. Dexamethasone (1 mg/kg of body weight/d) was administered intramuscularly to five irradiated animals every 24 hours, beginning 2 days before and ending 7 days after irradiation. Magnetic resonance imaging and proton magnetic resonance spectroscopy were performed 1 week, 1 month, and 3 months after irradiation. RESULTS: Magnetic resonance imaging performed 1 week after irradiation revealed marked edema in five nontreated animals. In dexamethasone-treated animals, the volume of edema was reduced significantly, compared to that of nontreated animals, 1 week and 1 month after irradiation. The volume of ring enhancement in dexamethasone-treated animals was also reduced significantly, compared to that of nontreated animals, 3 months after the irradiation. Proton magnetic resonance spectroscopy spectra revealed that N-acetylaspartate and choline peaks were reduced 1 week after irradiation in both groups. However, there were no statistically significant differences between the two groups at any time points. CONCLUSION: These results suggest that dexamethasone treatment may have an antiedema effect at an early stage and may modify subsequent development of vascular and inflammatory changes but may have no effect of preventing radiation-induced necrosis and the reduction of N-acetylaspartate after brachytherapy.

New method for the evaluation of the response of cultured cell lines to continuous low-dose-rate irradiation from encapsulated iridium-192 seeds with hyperthermia.

A new method was designed to investigate and evaluate the biological effectiveness of hyperthermia combined with continuous low-dose-rate irradiation (CLDRI) from encapsulated iridium-192 seed sources on glioma cells in vitro using the MTT assay. The system consists of 10 iridium seeds contained in a catheter bent into a circle, which is placed on a culture plate containing the cells. The effects of CLDRI and CLDRI combined with hyperthermia on a cultured rat glioma cell line (C-6) were studied. The number of surviving cells decreased as the total radiation dose increased. There was no significant difference in survival rates at dose rates of 0.1 Gy/hr and of 0.2 Gy/hr (p = 0.2811). An additive effect was observed in the cells treated with hyperthermia at 41 degrees C and 42 degrees C, combined with CLDRI, and synergistic effect between the two treatment modalities was observed at 43 degrees C. This new device is less expensive, easily reproducible, and can also be performed easily enough to examine a large number of samples in a short time period for sensitivity testing.

Steroidal saponins from the tubers of Dichelostemma multiflorum and their inhibitory activity on cyclic-AMP phosphodiesterase.

Phytochemical examination of the tubers of Dichelostemma multiflorum led to the isolation of three new steroidal saponins together with two known saponins. The structures of the new compounds were determined by spectral data and a few chemical transformations to be (25R)-5 alpha-spirostane- 1 beta,3 beta-diol (brisbagenin) 1-O-[O-alpha-L-rhamnopyranosyl-(1-->3)-4-O- acetyl-alpha-L-arabinopyranoside], brisgabenin 1-O-[O-alpha-L-rhamnopyranosyl-(1-->2)-O- [alpha-L-rhamnopyranosyl-(1-->3)]-4-O-acetyl-alpha-L-arabinopyranosid e] and (22S,25S)-5 alpha-spirostan-3 beta-o1 3-O-[O-beta-D-galactopyranosyl- (1-->2)-O-[beta-D-xylopyranosyl-(1-->3)]-O-beta-D-glucopyranosyl-(1-->4) -beta-D-galactopyranoside]. The known compounds were identified as desglucolanatigonin II and gitonin, with certain amounts of the corresponding C-25S isomers. Inhibitory activity of the isolated saponins and their derivatives on cAMP phosphodiesterase was evaluated to identify new compounds with medicinal potential.

Structures of steroidal saponins from the tubers of Brodiaea californica and their inhibitory activity on tumor promoter-induced phospholipid metabolism.

Phytochemical examination of the fresh tubers of Brodiaea californica resulted in the isolation of four new steroidal saponins. Their structures were determined, by extensive spectral analysis including two-dimensional (2D) NMR spectroscopy and acid-catalyzed hydrolysis, to be (25S)-spirost-5-ene-1 beta,3 beta-diol [(25S)-ryscogenin] 1-O-[O-beta-D-glucopyranosyl-(1-->3)-O-alpha-L-rhamnopyranosyl-(1-->2)- beta-D-glucopyranoside] (1), (25S)-ruscogenin 1-O-[O-beta-D-glucopyranosyl-(1-->3)-O-alpha-L-rhamnopyranosyl-(1-->2)-O -[beta-D-xylopyranosyl-(1-->3)]-beta-D-glucopyranoside] (2), the C-20 and C-22 isomer of 2 (3) and the 6'-O-acetyl derivative of 2 (4), respectively. The conformations of the tetrasaccharide moiety of 2 and 4 were inspected through molecular mechanics and molecular dynamics calculation studies, showing that the acetyl group attached to C-6 of the inner glucose was near the C-21 methyl of the aglycon in the calculated preferred conformation of 4, which must cause the downfield shift of 21-Me by 0.07 ppm in comparing the 1H-NMR of 4 with that of 2. The inhibitory activity of the isolated saponins on 12-O-tetradecanoylphorbor-13-acetate (TPA)-stimulated 32P-incorporation into phospholipids of HeLa cells was evaluated to identify new antitumor-promoter compounds.

[Studies on the constituents and biological activities of simaroubaceous plants].

In search of new compounds in simaroubaceous plants, we have been investigating naturally occurring substances [alkaloids, quassinoids, limonoids and picrotoxane-type terpenoids] on the following subjects: I. chemical studies on the constituents of Japanese simaroubaceous plants (I-1 and 2), II. chemical studies on the constituents of Indonesian and Chinese simaroubaceous plants (II-1-9), III. biological activities of the isolated natural compounds mainly and the chemical modification analogs (III-1-9). This article reviews the results obtained in our laboratory since 1976 on the subjects of I, II and III.

[Synergistic effect of carboplatin and hyperthermia in rat and human glioma cell lines].

To assess the interaction of carboplatin and hyperthermia in vitro, the thermochemosensitivities of three glioma cell lines, C6 rat glioma cell line, human glioma cell lines T98G and KMG4, were examined by 3-(4,5-dimethylthiazol-2-yl) -2,5 diphenyltetrazolium bromide (MTT) assay. The cell survival of each cell line decreased according to increasing CBDCA concentration and temperature. With a certain CBDCA concentration, the cell survival at following temperature was significantly decreased from that at 37 degrees: 43 degrees C and 44 degrees C for C6 cells (2.5 micrograms/ml); 41 degrees C, 42 degrees C, 43 degrees C and 44 degrees C for C6 cells (128 micrograms/ml); 42 degrees C, 43 degrees C and 44 degrees C for KMG4 cells (8 micrograms/ml) (CBDCA concentration within parentheses). It is generally considered that the highest tolerable temperature of normal brain is 42 degrees C for 60 minutes, while under 43 degrees C, there is a possibility that a sufficient tumoricidal effect might not be obtained. This study revealed enhanced cytotoxicity of CBDCA with hyperthermia at the temperature lower than 42 degrees C and suggests the possibility to gain increased tumoricidal effect without injuring normal brain by hyperthermia at the normal-tissue-tolerant temperature with systemic administration of relatively lower dose of CBDCA.

Triterpenoid saponins from Ardisia crenata.

Two novel triterpenoid saponins, ardisicrenoside A [3 beta-O-(alpha-L-rhamnopyranosyl-(1-->2)-[beta-D-glucopyranosyl- (1-->4)-[beta-D-glucopyranosyl-(1-->2)]-alpha-L-arabinopyranosyl)- 13 beta,28-epoxy-16 alpha,30-oleananediol] and ardisicrenoside B [3 beta-O-(beta-D-xylopyranosyl-(1-->2)-[beta-D- glucopyranosyl-(1-->4)-[beta-D-glucopyranosyl-(1-->2)]- alpha-L-arabinopyranosyl)-13 beta,28-epoxy-16 alpha,30-oleananediol] were isolated from the roots of Ardisia crenata. Two known triterpenoid saponins, ardisiacrispins A and B were also isolated from this source. Their structures were determined mainly by 2D NMR (COSY, HOHAHA, HETCOR, HMBC and ROESY) experiments. The aglycones are the new 13 beta,28-epoxy-3 beta,16 alpha,30-oleananetriol for ardisicrenosides A and B.

Triterpenoid saponins from Ardisia crenata and their inhibitory activity on cAMP phosphodiesterase.

Two novel triterpenoid saponins, ardisicrenoside C (1) [3 beta-O-(alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranosyl-(1-->4)- [beta-D-glucopyranosyl-(1-->2)]-alpha-L-arabinopyranosyl)-16 alpha, 28-dihydroxy-olean-12-en-30-oic acid 30-O-beta-D-glucopyranosyl ester] and ardisicrenoside D (2) [3 beta-O-(beta-D-xylopyranosyl-(1-->2)- beta-D-glucopyranosyl-(1-->4)-[beta-D-glucopyranosyl-(1-->2)]-alpha-L- arabinopyranosyl)-16 alpha, 28-dihydroxy-olean-12-en-30-oic acid 30-O-beta-D-glucopyranosyl ester] were isolated from the roots of Ardisia crenata. Structure assignments are based on spectroscopic data including 2D-NMR (correlation spectroscopy (COSY), homonuclear Hartmann-Hahn spectroscopy (HOHAHA), heteronuclear correlated spectroscopy (HETCOR), heteronuclear multiple bond correlation (HMBC) and rotating frame NOE spectroscopy (ROESY)) experiments and some chemical reactions. In addition, the isolated saponins along with their prosapogenins and sapogenins have been evaluated for their inhibitory activity on cAMP phosphodiesterase as a primary screening test for new medicinal compounds.

Acute effects of interstitial hyperthermia on normal monkey brain--magnetic resonance imaging appearance and effects on blood-brain barrier.

The magnitude and time course of histological and neuroradiological changes due to interstitial hyperthermia in normal cerebral white matter were investigated in eight adult Japanese monkeys. A cooling system enveloping a 2450-MHz microwave antenna was inserted stereotactically into the brain under general anesthesia. A point located 5 mm away from the surface of the cooling system was used as the reference point (RP). Hyperthermia was given to maintain the RP at 43 degrees C for 60 minutes. Two animals were sacrificed under general anesthesia following the intravenous administration of Evans blue, immediately and 1, 3, and 7 days after treatment. After removing the brain, histological changes were investigated. Magnetic resonance (MR) imaging was performed at 1, 3, and 7 days after treatment. Evans blue extravasation was most prominent in the region heated to 43 degrees C or above immediately after treatment. MR imaging showed obvious enhancement in the region heated to 43 degrees C or above 1 day after treatment, related to the disruption of the blood-brain barrier (BBB) by hyperthermia. Three days after treatment, ring-like enhancement with a central low-intensity area was seen in the region heated to about 43 degrees C, caused by BBB disruption and slight neovascularization. One week after treatment, an enhanced ring was observed in the region heated to less then 43 degrees C which surrounded a low-intensity area. The enhancement seen 1 week after treatment was caused by prominent neovascularization. T2-weighted imaging showed a high-intensity area, caused by edema, most prominent 3 days after treatment. Thus chemotherapeutic agents should be given just before the end of hyperthermia.

Canthin-6-one alkaloids from Brucea mollis var. tonkinensis.

Three new alkaloids were isolated from the root-wood of Brucea mollis var. tonkinensis collected in China. Their structures were determined to be 11-O-beta-D-glucopyranosyl-(1-->6)-beta- D-glucopyranosylcanthin-6-one, 5-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosylcanthin-6-o ne and 11-hydroxycanthin-6-one-N-oxide, by chemical and spectral methods. In addition, two known alkaloids, canthin-6-one and canthin-6-one-N-oxide, were isolated.

Studies on the constituents of Ailanthus integrifolia.

A new phenolic glycoside, 3,4,5-trimethoxyphenol-1-(6-xylopyranosyl)glucopyranoside, was isolated together with twenty known compounds identified as koaburaside, 3,4,5-trimethoxyphenol, 5,7-dihydroxychromone-7-neohesperidoside, naringin, neoeriocitrin, p-coumaric acid, vanillin, vanillic acid, coniferyl aldehyde, ferulic acid, trans-triacontyl-4-hydroxy-3-methoxycinnamate, p-methoxycinnamic acid, 2,6-dimethoxybenzoquinone, 2-(1-hydroxyethyl)naphtho[2,3-b]furan-4,9-dione, 2-acetylnaphtho[2,3-b]furan-4,9-dione, 2-(1-hydroxyethyl)-6-methoxynaphtho[2,3-b]furan-4,9-dione, 2-acetyl-6-methoxynaphtho[2,3-b]furan-4,9-dione, specioside, jioglutin C and rehmaglutin D from the bark of Ailanthus integrifolia Lamk (Simaroubaceae).

Three new furostanol saponins from the bulbs of Ipheion uniflorum.

Phytochemical screening of the bulbs of Ipheion uniflorum (Liliaceae) has led to the isolation of three new furostanol saponins (2-4) along with a known phytoecdysteroid, ecdysterone (1). The structures of the new compounds were determined by two-dimensional NMR techniques, 1H-1H correlation spectroscopy (COSY), homonuclear Hartmann-Hahn (HOHAHA), phase-sensitive nuclear Overhauser effect spectroscopy (NOESY), heteronuclear multiple quantum coherence (HMQC) and heteronuclear multiple-bond correlation (HMBC) spectra, and hydrolysis to be 3 beta-hydroxy-22 alpha-methoxy-26-O-beta-D- glucopyranosyloxy-5 alpha-furost-25(27)-en-2-one 3-O-[O-alpha-L-rhamnopyranosyl-(1-->2)-O-[O-alpha-L-arabinopyranosyl+ ++- (1-->2)-O-[beta-D-xylopyranosyl-(1-->3)]-beta-D-glucopyranosyl-(1-->4)]- beta-D-galactopyranoside) (2) and its 25(27)-dihydro derivatives (3: 25S; 4: 25R). The inhibitory activity exhibited by compounds 1-4 and the corresponding spirostanol saponins (2a and 3a) of 2 and 3 on cAMP phosphodiesterase was assayed as a primary screening test to identify new compounds with medicinal potential.