Differences between cytotoxicity in photodynamic therapy using a pulsed laser and a continuous wave laser: study of oxygen consumption and photobleaching.

Oxygen consumption at the targeted site has a significant effect on dosimetry in photodynamic therapy (PDT). However, oxygen consumption in PDT using a pulsed laser as a light source has not been clarified. We therefore investigated the dependence of cytotoxicity on the oxygen consumption and the photosensitizer photobleaching of PDT using a pulsed laser by comparing with that using a continuous wave (CW) laser. Mouse renal carcinoma cells (Renca) were incubated with a second-generation photosensitizer, PAD-S31. The cells were then irradiated with either a 670-nm nanosecond pulsed light from the 3rd harmonics of a Nd:YAG laser-pumped optical parametric oscillator with a peak fluence rate of approximately 1 MW/cm(2) at 30 Hz or a 670-nm CW diode laser with a total light dose of 40 J/cm(2). Regardless of laser source, cytotoxic effects exhibited cumulative dose responses to the photosensitizer ranging from 12 to 96 microg/ml. However, cytotoxic effect of PDT using the pulsed light was significantly less than that using the CW light with the photosensitizer concentrations of 24 and 48 microg/ml under identical fluence rates. During PDT, the cells exposed to the pulsed light consumed oxygen more slowly, resulting in a lower amount of oxygen consumption when compared with PDT using CW light. In accordance with oxygen consumption, the pulsed light induced significantly less photobleaching of the photosensitizer than the CW light did. These results indicate that the efficiency of PDT using pulsed light is less when compared with CW light, probably being related to suppressed oxygen consumption during the pulsed light irradiation.

Treatment planning for capacitive regional hyperthermia.

Capacitively coupled hyperthermia devices are widely in use, mainly in Asian countries. In this paper, a comprehensive treatment planning system, including a Specific Absorption Rate (SAR) and thermal model for capacitively coupled hyperthermia, is described and demonstrated using a heterogeneous patient model. In order to accurately model a hyperthermia treatment, simulation at high resolution is mandatory. Using the quasi-static approximation, the electromagnetic problem can be solved at high resolution with acceptable computational effort. The validity of the quasi-static approximation is demonstrated by comparing the Maxwell solution of a phantom problem to the quasi-static approximation. Modelling of capacitive hyperthermia of the prostate reveals the difficulty of heating deep-seated tumours in the pelvic area. Comparison of the SAR distribution in the heterogeneous patient model and a patient shaped agar phantom shows a shielding effect of the pelvic bone and the influence of the fat-muscle distribution. It is shown that evaluation of capacitive hyperthermia with agar phantoms leads to overly optimistic conclusions. Therapeutic relevant tumour temperatures can only be obtained by permitting temperature extrema in normal tissue. This concurs with clinical practice, where treatment-limiting hot spots restrict the tumour temperature. It is demonstrated that the use of very cold overlay bolus bags has only a very superficial effect. The presented model can be used for individual treatment planning and optimization, for the evaluation of capacitive applicator modifications and comparison with other devices.

Safety evaluation of proanthocyanidin-rich extract from grape seeds.

Proanthocyanidins, extracted from grape seeds, are widely used mainly as nutritional supplements. However, there has not been a systematic report to investigate toxicological studies on proanthocyanidins, especially in oral administration. In our studies, proanthocyanidin-rich extract from grape seeds was subjected to a series of toxicological tests to document its safety for use in various foods. The grape seed extract (GSE) was examined for acute and subchronic oral toxicity using Fischer 344 rats and for mutagenic potential by the reverse mutation test using Salmonella typhimurium, the chromosomal aberration test using CHL cells, and the micronucleus test using ddY mice. No evidence of acute oral toxicity at dosages of 2 and 4 g/kg, and no evidence of mutagenicity in the above tests was found. Administration of GSE as a dietary admixture at levels of 0.02, 0.2 and 2% (w/w) to the rats for 90 days did not induce noticeable signs of toxicity. The no-observed-adverse-effect level (NOAEL) of GSE in the subchronic toxicity study was 2% in the diet (equal to 1410 mg/kg body weight/day in males and 1501 mg/kg body weight/day in females). The results of our studies indicate a lack of toxicity and support the use of proanthocyanidin-rich extract from grape seeds for various foods.

Low power diode laser treatment using indocyanine green for eradication of esophageal varices.

BACKGROUND AND STUDY AIMS: Endoscopic variceal ligation (EVL) is an alternative to sclerotherapy for the treatment of esophageal varices, but is associated with higher rates of recurrence and subsequent bleeding than sclerotherapy. To prevent recurrence of varices after EVL, we have developed a low-dose diode laser therapy combined with the injection of indocyanine green, which allows enhanced tissue absorption of the laser beam selectively around varices. In this study we investigated the efficacy and safety of this technique. PATIENTS AND METHODS: Eight patients with F2 or F3 esophageal varices were enrolled. At 1 week after EVL, indocyanine green solution (1 mg/ml) was injected submucosally around the remaining varices. A diode laser (power 10 watts) was applied to the surface from the esophagogastric junction to 5 cm above it. The spot size was kept to 5 mm in diameter. RESULTS: Laser irradiation was performed safely, without bleeding from the varices, or perforation. There were no major complications. Endoscopy 1 month later showed F0 forms in seven patients, F1 in one patient, and no red color sign in any patient. No recurrence of varices has been observed in any of the patients during the follow-up period of at least 12 months. CONCLUSION: This technique may provide a simple, safe and effective procedure, as an additional treatment to EVL, for the prevention of recurrence of esophageal varices.

Structures of new sesquiterpenoids from Farfarae Flos.

Two new bisabolane-type sesquiterpenoids, (3R,4R,6S)-3,4-epoxybisabola-7(14),10-dien-2-one and (1R,3R,4R,5S,6S)-1-acetoxy-8-angeloyloxy-3,4-epoxy-5-hydroxybisabola-7(14),10-dien-2-one, and a new oplopane-type sesquiterpenoid, 14(R)-hydroxy-7beta-isovaleroyloxyoplop-8(10)-en-2-one, were isolated from Farfarae Flos along with three known compounds. The structures of these compounds were elucidated on the basis of spectroscopic evidence.

Secoiridoid glycosides from Gentiana scabra.

Two new secoiridoid glycosides, 4'-O-beta-D-glucopyranosylgentiopicroside (1) and 6'-O-beta-D-glucopyranosylgentiopicroside (2), have been isolated from the rhizomes and roots of Gentiana scabra together with three known compounds, olivieroside, 1-O-beta-D-glucopyranosylamplexine, and benzyl alcohol O-alpha-L-arabinopyranosyl (1 --> 6)-beta-D-glucopyranoside. The structures of 1 and 2 were elucidated using chemical and physicochemical (MS and NMR) studies.

Isoflavone aglycone-rich extract without soy protein attenuates atherosclerosis development in cholesterol-fed rabbits.

The antiatherogenic effect of soy protein with intact isoflavones is well established, but the effects of isoflavones without soy protein have not been determined. We investigated the antiatherogenic effect of an isoflavone aglycone-rich extract (containing 429.4 mg/g isoflavone aglycones) without soy protein from fermented soy in cholesterol-fed rabbits. We fed 12-wk-old New Zealand white male rabbits diets containing 1 g/100 g cholesterol with 0, 0.33 or 1 g/100 g isoflavone aglycones for 8 wk. We also fed the rabbits a diet containing 1 g/100 g cholesterol with 1.09 g/100 g soy saponin-rich extract, a component other than isoflavone aglycones in the isoflavone aglycone-rich extract. Controls did not consume cholesterol, isoflavone aglycones or saponins. The isoflavone aglycone- and saponin-rich extracts did not affect the serum lipid profile of cholesterol-fed rabbits. The serum concentration of daidzein in its conjugated form was significantly higher in the high isoflavone group than in the low isoflavone group. The level of cholesteryl ester hydroperoxide (ChE-OOH) induced by CuSO(4) in plasma in the high isoflavone group was significantly less than that in the cholesterol group, and the ChE-OOH levels of LDL in the low and high isoflavone groups were significantly less than those in the cholesterol group. The ChE-OOH levels in plasma and LDL in the saponin group did not differ from the cholesterol group. In the aortic arch, the cholesterol concentration was significantly lower in the high isoflavone group, and malondialdehyde concentration was significantly lower in the low and high isoflavone groups compared with the cholesterol group; however these concentrations in the saponin group did not differ from those in the cholesterol group. The atherosclerotic lesion area of the aortic arch was significantly lower in the isoflavone groups (26.3% lower in the low isoflavone group and 36.9% lower in the high isoflavone group) than in the cholesterol group. The lesion areas were not different in the soy saponin and cholesterol groups. Immunohistochemical analysis revealed fewer oxidized LDL-positive macrophage-derived foam cells in atherosclerotic lesions in the aortic arch of isoflavone groups compared with that of the cholesterol group. These results suggest that the antioxidative action of isoflavones and their antioxidative metabolites inhibit the oxidation of LDL, thereby exerting an antiatherosclerotic effect.

[A case of recurrent breast cancer with carcinoma erysipeloides responding to sequential therapy with docetaxel (TXT) and doxifluridine (5'-DFUR) accompanied by leucovorin (LV)].

A left radical mastectomy was performed on a 53-year-old woman, diagnosed with left inflammatory breast cancer, after local arterial chemotherapy with cyclophosphamide (CPA), doxorubicin and 5-fluorouracil (5-FU). Adjuvant therapy was added with irradiation and ECF. Four months after the operation, a red eruption was detected on the left upper chest wall. The lesion was diagnosed by skin biopsy as a recurrent breast cancer with carcinoma erysipeloides. Tumor marker levels suggested the recurrent cancer was ECF resistant, so we changed the chemotherapy regime to a single dose of TXT. Although tumor marker levels and the skin eruptions improved at the beginning of the therapy, pleuritis carcinomatosa was found. We changed the regime again to a continuous dose of 5'-DFUR and LV for day 1 to 7. With this regime the clinical symptoms improved, and 2 courses of this modified FL therapy were carried out. After the therapy, the tumor seemed resistant to this modified FL therapy. Therefore, we tried a sequential therapy with TXT and the modified FL, which induced an improvement in clinical symptoms. Two years later, the patient died from the breast cancer. Therefore, we conclude that the sequential therapy may be beneficial in managing untreatable carcinoma erysipeloides of recurrent breast cancer.

Studies of the constituents of Gardenia species. II. Terpenoids from Gardeniae Fructus.

Four new terpenoids, gardenate A (1), 2-hydroxyethyl gardenamide A (2), (1R,7R,8S,10R)-7,8,11-trihydroxyguai-4-en-3-one 8-O-beta-D-glucopyranoside (3) and Jasminoside F (4), were isolated from Gardeniae Fructus. Their structures were established on the basis of spectral analysis.

Protective effect of aged garlic extract on the small intestinal damage of rats induced by methotrexate administration.

The methotrexate (MTX) administration to rats causes the damage of small intestine. The small intestinal damage was evaluated by measuring the intestinal permeability of the poorly absorbable compound, fluorescein isothiocyanate (FITC)-labeled dextran (average molecular weight, 4,400) (FD-4) using the in vitro everted intestine technique and by determining the FD-4 that appeared in plasma using the in situ closed loop intestine technique. The MTX administration to rats fed with the standard laboratory diet increased the small intestinal permeability of FD-4 due to the damage of the small intestine. Interestingly, the permeability of FD-4, when MTX was administered to rats fed with the aged garlic extract containing diet, was depressed almost to the level of control rats without the MTX treatment. The present study showed that the aged garlic extract protected the small intestine from the damage induced by the action of MTX on the crypt cells.

Atrial natriuretic peptide in aged patients with iron deficiency anemia.

Anemia is a common disease in elderly people. However, since hemoglobin concentration often decreases subclinically with aging because of nutritional impairment, its pathological significance is unclear. To investigate the pathological significance of low hemoglobin concentration, we studied the relation between hemoglobin levels and arrhythmia, as well as circulatory parameters. Arrhythmia was detected by Holter type ambulatory electrocardiography in 42 elderly people (aged 60 or over) living in a nursing home. Plasma concentrations of human atrial natriuretic peptide (hANP) after iron therapy were determined by immunoradiometric assay. Changes in circulatory parameters in elderly people with iron deficiency anemia were examined. Supraventricular and ventricular premature contractions significantly increased in elderly people with low hemoglobin concentrations or hematocrit. hANP increased significantly as the hemoglobin concentration decreased in 22 elderly people. Of these 22 subjects, 11 showed a low serum concentration of iron, and were administered ferrous salts. No side effects, such as nausea, occurred. After iron supplementation, the average hemoglobin level increased from 9.0 to 10.5 g/dl, and the average hANP level was reduced from 58.3+/-23.5 to 41.2+/-27.9 pg/ml, which was statistically significant by Wilcoxon's signed rank sum test. The increase in the hemoglobin level inversely correlated with the hANP level. Heart rate, blood pressure and body weight of subjects decreased significantly after iron supplementation therapy. Although hemoglobin levels were increased by iron supplementation therapy after a long period of anemia, the duration of the period with low hemoglobin levels showed no significant relation to initial hANP concentration. In conclusion, low hemoglobin levels induced secretion of hANP, and treatment of iron deficiency might exert favorable effects on the circulatory system.

Simultaneous determination of nicotinic acid and its metabolites in rat urine by micellar electrokinetic chromatography with photodiode array detection.

Nicotinic acid, nicotinamide and their possible metabolites were successfully separated within 17 min by micellar electrokinetic chromatography using 50 mM borate buffer (pH 9.0) containing 150 mM sodium dodecyl sulfate as the running buffer. Calibration curves for all compounds showed good linearity in a range of 5 microg/ml and 250 microg/ml with good correlation. The present method did not require any clean-up procedures and made it possible to determine all metabolites without interference on a photodiode array detector. Urine samples collected from Wistar male rats were analyzed after high-dose oral or intravenous administration of nicotinic acid or nicotinamide. Metabolic pathways of nicotinic acid in male Wistar rats are also discussed.

Cerebrosides A and C, sphingolipid elicitors of hypersensitive cell death and phytoalexin accumulation in rice plants.

When plants interact with certain pathogens, they protect themselves by generating various chemical and physical barriers called the hypersensitive response. These barriers are induced by molecules called elicitors that are produced by pathogens. In the present study, the most active elicitors of the hypersensitive response in rice were isolated from the rice pathogenic fungus Magnaporthe grisea, and their structures were identified as cerebrosides A and C, sphingolipids that were previously isolated as inducers of cell differentiation in the fungus Schizophyllum commune. Treatment of rice leaves with cerebroside A induced the accumulation of antimicrobial compounds (phytoalexins), cell death, and increased resistance to subsequent infection by compatible pathogens. The degradation products of cerebroside A (fatty acid methyl ester, sphingoid base, and glucosyl sphingoid base) showed no elicitor activity. Hydrogenation of the 8E-double bond in the sphingoid base moiety or the 3E-double bond in the fatty acid moiety of cerebroside A did not alter the elicitor activity, whereas hydrogenation of the 4E-double bond in the sphingoid base moiety led to a 12-fold decrease in elicitor activity. Furthermore, glucocerebrosides from Gaucher's spleen consisting of (E)-4-sphingenine and cerebrosides from rice bran mainly consisting of (4E,8E)-4,8-sphingadienine and (4E,8Z)-4,8-sphingadienine showed no elicitor activity. These results indicate that the methyl group at C-9 and the 4E-double bond in the sphingoid base moiety of cerebrosides A and C are the key elements determining the elicitor activity of these compounds. This study is the first to show that sphingolipids have elicitor activity in plants.

[Myotonic dystrophy with marked megacolon: report of a case].

A 45-year-old woman was incidentally suspected to have megacolon. Chest X-rays showed elevated left diaphragm due to colonic gas, and the heart was deviated to the midline. Barium enema revealed marked dilation of the sigmoid colon, confirming the diagnosis of megacolon. Maximal diameter of the sigmoid colon was 23 cm, but she had no gastrointestinal symptoms. During the work up for megacolon, the presence of myotonic dystrophy was suspected. She had hatchet face, but was not bald. Muscles of the neck and extremities were slightly atrophic. There was percussion myotonia of the tongue and both hands, and grip myotonia of the hands. Laboratory examinations showed impaired glucose tolerance and low level of serum IgG. EMG showed myotonic discharges and myopathic units in the limbs. Brain CT imaging revealed a thick skull. Cases of myotonic dystrophy associated with marked megacolon are rare in Japan. Megacolon presents a high risk for ileus, volvulus, and rupture, and myotonic dystrophy is associated with a high operative and anesthesic risk. Megacolon, therefore, is an important complication to look for in the management of myotonic dystrophy.

The development of a high impedance all ceramic thermistor resistant to microwave irradiation: its possible medical application.

We developed a prototype all ceramic thermistor (ACT) composed of a silicon carbide (SiC) ceramic thermistor and SiC ceramic fibre-lead wires in order to accurately measure tissue temperature during microwave heating treatment. High density SiC ceramics reflects electromagnetic waves and manifests microwave resistant properties. The resistance of the SiC ceramic thermistor is approximately 15 M Ohm at 20 degrees C and has a sensitivity of approximately 200 K Ohm/ degrees C. SiC ceramic fibre resistance is approximately 40 K Ohm/cm, whereas its sensitivity is approximately 10 Ohm/ degrees C-cm, significantly lower than that of the SiC ceramic thermistor. The resistance of the SiC ceramic fibre is stable. This enables the ACT to be designed with the two-probe method instead of the four-probe method which was adopted in the Bowman type thermistor, resulting in a greatly simplified model. The measurement error caused by the temperature dependence of SiC ceramic fibre is < 0.02 degrees C. The ACT was inserted into muscle-equivalent agar phantom and into gluteal muscles of rabbits, and microwave irradiation was carried out. We recorded the phantom and tissue temperature without disturbance by the electromagnetic field for heating. This prototype ACT appears promising for its possible clinical applications in such settings as hyperthermic cancer therapy in man.

[Anemia and neutropenia in elderly patients caused by copper deficiency for long-term enteral nutrition].

Anemia and neutropenia caused by copper deficiency is a well-known consequence of long term total parenteral nutrition in the literature. We present 6 bed-ridden elderly patients who developed anemia and neutropenia after receiving enteral nutrition for a long time (mean: 3.3 years) In all 6 patients, serum copper and ceruloplasmin level were very low, and the mean of their hematological data were as follows: WBC 2,200/microliters, neutrophil 554/microliters, hemoglobin 8.1 g/dl, platelet 260 x 10(3)/microliters, respectively. The bone marrow examination showed cytoplasmic vacuolization of both myeloid and erythroid precursors, and maturation arrest of granulopoiesis. Then, copper sulfate was administrated by enteral tube to 6 patients, and the improvement of anemia and neutropenia was observed within a month. A 82-year-old woman who received enteral nutrition for 3.5 years with sever anemia (Hb 3.7 g/dl) and neutropenia (neutrophil 350/microliters), showed a marked improvement in hematological data (Hb 8.0 g/dl, neutrophil 4, 092/microliters, respectively) after two months by administering the copper supplementation. The exact cause of the anemia and neutropenia in copper deficiency is unclear, but it is suggested that the decreased activity of enzyme containing copper may be related. Hematological abnormalities due to copper deficiency should be cared during long term enteral nutrition with long termed bed-ridden elderly patients.

Hepatocellular carcinoma after transcatheter hepatic arterial embolization. A histopathologic study of 84 resected cases.

BACKGROUND: Recently, transcatheter arterial embolization (TAE) has been used to treat hepatocellular carcinoma (HCC), yet much is still unknown regarding its optimal use. METHODS: Eighty-four patients with HCC after TAE underwent surgical resection. Fifty of the tumors were less than 3 cm (small HCC [S-HCC]), and 34 were 3 cm or larger (large HCC [L-HCC]). Necrosis rate, distribution of residual HCC, histopathology of the main tumor, and proliferating activity of residual HCC by means of proliferative cell nuclear antigen (PCNA) were examined. Twenty-two randomly selected patients with HCC treated with standard chemotherapy were used as non-TAE control subjects. RESULTS: A necrosis rate of greater than 95% was seen in 35 cases of S-HCC and in 15 of L-HCC. All five nonencapsulated tumors were L-HCC and had a much lower necrosis rate. No tumors in the control group showed a necrosis rate of greater than 95%. Encapsulated tumors were categorized according to their tumor interiors, capsules, and extracapsular zones. Complete necrosis of the tumor interior was 80.0% and 35.3% in S-HCC and L-HCC, respectively. Viable residual tumors were found mainly in the extracapsular zone in S-HCC, whereas in L-HCC they were located primarily in the tumor interior. Most capsules were affected by tumor necrosis and the subsequent healing process, resulting in a thick secondary capsule. Tumor interior necrosis was uniform and coagulative in S-HCC, in contrast to L-HCC, in which necrotic regions comprised several necrosis units of differing texture and were divided by fibrous septa. In contrast, the control group revealed spotty, sparse necrosis. Non-TAE tumor capsules were thin and pathologically characteristic of those naturally occurring in tumors, as opposed to the thick fibrous capsules, which are inducible by TAE therapy. In the TAE group, the PCNA positivity rates were 37.5%, 52.5%, and 100% in Grades 1, 2, and 3/4 combined, respectively. At the tumor-nontumor boundary of the extracapsular region, PCNA-positive cells were detected in 55.0% of the cases. CONCLUSIONS: The thickened tumor capsule serves as a good postoperative indicator of TAE response. Small tumors seem to be affected in the tumor interior, whereas extracapsular invasion undermines the TAE effect. PCNA was helpful in detecting the tumor-nontumor boundary and useful as a parameter of viability of HCC after TAE.

Contact hemolysin production by strains of enteroaggregative Escherichia coli isolated from children with diarrhea.

Forty-five strains of enteroaggregative Escherichia coli were tested for hemolytic activity with different culture media and erythrocytes from different species. Thirty-seven strains showed proteinase K-sensitive contact hemolysin activity with sheep erythrocytes when cultured in Casamino Acids-yeast extract broth supplemented with 1 mM calcium chloride and when cultured in nutrient broth media. The production of contact hemolysin was dependent on temperature, pH, and culture conditions.

Clostridial gas gangrene associated with congenital generalized lipodystrophy: report of a case.

Congenital generalized lipodystrophy (CGL) is a rare disease, the main symptoms of which are a reduction of systemic subcutaneous fat, characteristic facial features, hypertrichosis, and insulin-resistant diabetes. We report herein the unusual case of a 25-year-old man with CGL in whom gas gangrene developed, an association which has never before been encountered.