Clinical pilot studies on pre-operative hyperthermic tumour ablation for advanced breast carcinoma using an 8 MHz radiofrequency heating device.

The multimodality treatment approach for advanced breast cancer provides survival advantages with decreased locoregional and distant recurrences, but these intensive anti-tumour treatments cause severe myelosuppression. Thus, in this study, the usefulness of pre-operative anti-tumour treatment without myelosuppression was investigated. Nine patients with advanced breast carcinoma underwent pre-operative hyperthermic tumour ablation (HTA) using an 8 MHz radiofrequency (RF) heating device (Thermotron RF-8) combined with a grounded needle electrode. The patients had a mean age of 58.3+/-13.9 years and included four patients with stage IIIA, two with stage IIIB and three with stage IV cancer. The target temperature was over 50 degrees C. They tolerated pre-operative HTA therapy well with no early or late complications. The initial mean tumour size was 122.1+/-71.5 cm3 and the post-HTA tumour size was 82.2+/-63.4 cm3; the reduction rate was significant (p = 0.000 293). After the pre-operative HTA, all patients underwent surgery with Level III nodal extirpation. Post-operatively, no locoregional recurrence was observed. Microscopic examination of the primary focus showed complete coagulation necrosis expanding for a diameter of 3.5-5.0 cm. Taken together, the pre-operative HTA was a safe, well-tolerated and effective treatment, achieving tumour reduction as well as complete coagulation necrosis that resulted in a large volume of destruction in breast cancer tissue.

Successful hyperbaric oxygen treatment of life-threatening hemorrhagic cystitis after allogeneic bone marrow transplantation.

Hemorrhagic cystitis (HC) is a major cause of morbidity after allogeneic bone marrow transplantation (BMT). Many therapies have been investigated to prevent or treat HC, but effective treatment for HC is still limited. While the efficacy of hyperbaric oxygen therapy has been established for HC due to chemotherapy and/or radiation therapy, its role in HC occurring after allogeneic BMT has yet to be defined. We report two cases of life-threatening late-onset HC after allogeneic BMT in children, which resolved after treatment with hyperbaric oxygen.

Development of a new heating needle for interstitial hyperthermia compatible with interstitial radiotherapy.

PURPOSE: We have developed a new heating needle for interstitial hyperthermia compatible with brachytherapy. We studied the heating characteristics of interstitial needles and the usefulness of these needles for interstitial hyperthermia. MATERIALS AND METHODS: As heating needles, we used MicroSelectron HDR interstitial needles. These needles were only heated at the metal tip, and were insulated in the middle and grounded from the top. The hyperthermia system was based on the principles of RF capacitive heating, and we used a Thermotron RF-8. We examined whether the temperature and the heating area could be elevated to a level required for hyperthermia. RESULT: Good heating was obtained around the metal tip of the needle. The heating area of the longitudinal axis of the needle varied with the length of the metal part, while the transverse heating area of the needle was almost unchanged. If heating was performed while maintaining the elevation of temperature in each needle at the same level using variable coils (tuning box), heating was homogeneous over the regions in accordance with the arrangement of the needles. CONCLUSION: With this heating system, the temperature rises easily, and a uniform temperature distribution can be obtained.

Effects of rooibos tea extract on antigen-specific antibody production and cytokine generation in vitro and in vivo.

Rooibos tea contains a large amount of flavonoids and acts as a potent antioxidant. In this study, we examined the effects of Rooibos tea extract on antigen-specific antibody production and cytokine generation in vitro and in vivo. The primary in vitro anti-ovalbumin (anti-OVA) or sheep red blood cell (SRBC) antibody production in murine splenocytes was markedly stimulated by the addition of the tea extract at concentrations of 1-100 microg/ml. On the other hand, a nonspecific antibody response elicited with lipopolysaccharide (LPS) in purified splenic B-cells was not modified by the extract. Rooibos tea extract caused an increase in the generation of interleukin 2 (IL-2) both in OVA- and anti-CD3-primed splenocytes at concentrations ranging from 10 microg/ml to 1000 microg/ml. In contrast, this tea extract suppressed the generation of interleukin 4 (IL-4) in OVA-primed splenocytes. Moreover, the reduction of OVA-induced antibody production in serum of the cyclosporin A (CyA) -treated rats can be significantly restored and the IL-2 generation in murine splenocytes was stimulated, following oral administrations of Rooibos tea extract. Thus, our findings suggested that Rooibos tea extract may facilitate the antigen-specific antibody production through selective augmentation of IL-2 generation both in vitro and in vivo. Collectively, Rooibos tea intake may be of value in prophylaxis of the diseases involving a severe defect in Th1 immune response such as cancer, allergy, AIDS, and other infections.

[Assessment of functional severity on in vivo hepatic 31P-MRS in diffuse hepatic disease: comparative studies with 99mTc-GSA].

The purpose of this study was to compare the assessment of functional severity on in-vivo hepatic 31P-MRS in diffuse hepatic disease with functional severity assessed with 99mTc galactosyl serum albumin (99mTc-GSA). 31P-MRS was performed in 10 healthy control subjects and 16 patients with diffuse hepatic disease. Data were expressed as peak area ratios: PME/beta-ATP, PDE/beta-ATP, PME/PDE, Pi/beta-ATP, and PME/Pi. The functional severity of hepatic damage was evaluated visually and quantitatively (HH15, LHL15) by 99mTc-GSA in the group of patients with diffuse hepatic disease. Visual evaluation was classified into four grades based on anterior images of cardiac blood-pool and liver. We studied the correlation of spectral metabolic ratios and functional severity by 99mTc-GSA. We found statistically significant differences (ANOVA) among the classifications of Grade I, Grade II, and Grade III with both PME/beta-ATP and PME/PDE. A statistically significant direct correlation was found between HH15 and both PME/beta-ATP and PME/PDE. A significant inverse correlation was also seen between LHL15 and both PME/beta-ATP and PME/PDE. The studies comparing 31P-MRS with functional severity assessed by 99mTc-GSA showed that PME/beta-ATP and PME/PDE were useful for the assessment of functional severity in patients with diffuse hepatic disease.

Indomethacin reduces the skin thermal damage in hyperthermic treatment of experimental malignant tumors.

Indomethacin (Ind), an inhibitor of prostaglandin synthesis, was previously shown to increase the thermosensitivity of murine tumors. The potency of drug to modify the thermal response of murine skin has been evaluated in mice heated in water bath at 44 degrees C for 30, 60 and 90 min. Ind was administered subcutaneously (s.c.) at dose of 5 mg/kg body weight (b.w.) 1.5 h before heating. The mouse foot skin reactions (FSR) were assessed using the scoring system of Urano et al (1979). The severity of skin thermal damage was decreased markedly by Ind. At the time when heating of the control group at 44 degrees C for 60 min resulted in the irreversible FSR in some mice, pretreatment with Ind before heating lead to the complete recovery from the heat damage in all mice. Similarly, after heating at 44 degrees C for 90 min, the degree of FSR was diminished by Ind from score 4.5 to score 2. It is concluded that Ind selectively protected normal skin during the hyperthermic treatment. Further clinical study is warranted.

Effects of muscle relaxants on EEG, ABR and EMG in rabbits.

The effects of baclofen and pancuronium bromide on evoked electromyogram (EMG), cortical electroencephalogram (EEG) and auditory brainstem responses (ABR) were studied in pentobarbital anesthetized normal rabbits. Evoked EMG was measured in the gastrocnemius muscle by electrical stimulation of the sciatic nerve. Intravenous injection of baclofen decreased EEG and arterial blood pressure and light reflex, however, it had no significant influence on EMG or ABR at doses of 10 and 20 mg/kg/h. Pancuronium bromide immediately inhibited respiration, decreased EEG and EMG, however, it had no significant influence on arterial blood pressure, ABR, or light reflex, at doses of 0.4 and 1.0 mg/kg/h in anesthetized rabbits. ABR waves were observed until just before cardiac arrest with both of the muscle relaxants. It is suggested that ABR are not influenced by central or peripheral muscle relaxants, or by pentobarbital.

Relevance of a new impedance matching, or subtrap, method for the reduction of pain during hyperthermia.

Capacitive heating is widely used in hyperthermic treatment of human malignancies. However, the pain on the body surface or thermoesthesia in the subcutaneous fatty layer may prevent an elevation of temperature in the tumors. Impedance matching is improved by a subtrap method entailing the application of two copper plates (10 x 850 x 0.06 mm) as a subtrap circuit to each of two capacitive electrodes. In a clinical trial the Tmax, Tave, Tmin for the subtrap method were all higher in comparison with those for the conventional technique (42.5 +/- 0.7 degrees C, 41.9 +/- 1.0 degrees C, 41.3 +/- 1.1 degrees C vs. 41.1 +/- 1.5 degrees C, 40.6 +/- 1.3 degrees C, 40.0 +/- 1.3 degrees C). Although the maximal radiofrequency (RF) power applied to patients was higher with the subtrap method (875 +/- 189 W vs. 763 +/- 200 W), the incidence of surface pain was reduced dramatically. It is concluded that the subtrap method substantially improves the RF capacitive heating of deep-seated tumors.

Characterization of a cDNA encoding CuZn-superoxide dismutase from the liverwort Marchantia paleacea var. diptera.

Suspension-cultured cells of the liverwort Marchantia paleacea var. diptera contain a cytosolic CuZn-superoxide dismutase (SOD) whose N-terminal amino acid sequence is similar to those of the isozymes found in chloroplasts of higher plants [Tanaka et al. (1996) Plant Cell Physiol. 37: 523]. A cDNA (MSODCc) encoding the cytosolic CuZn-SOD was isolated from cDNA library constructed from a liverwort cell suspension culture. The deduced amino acid sequence showed a higher degree of homology with the sequences of CuZn-SODs in chloroplasts than those in the cytosol of higher plants and an unique additional peptide in the C-terminal region, but no plastid transit sequence. Northern blotting using MSODCc as a probe and immunoblot analysis with antiserum against the enzyme revealed that the steady state level of transcript was not affected by copper, but both CuZn-SOD protein and its activity increased.

[Comparative studies on the antitumor activities and side effects of segmental SMANCS/Lip-TAE with segmental SMANCS/Lip-TAI for hepatocellular carcinoma].

We compared the effectiveness of treatments and the influence of side effects on liver function and clinical symptoms between segmental SMANCS/ Lip TAI and segmental SMANCS/Lip-TAE. The early tumor response rate of the group treated by TAI was 23.6%, and that of the group treated by TAE was 80.0%. In the group treated by TAE, the therapeutic effects were better in the nodular type than in the diffuse type of HCC, and we were also able to obtain a good tumor response rate on the multiple HCC and large HCC. However, there was no difference in the response period between the groups treated by TAI and TAE. In both groups, there were no significant differences in the appearance rate and degree of side effects. In conclusion, segmental SMANCS/Lip-TAE seemed to be an effective treatment for HCC without any serious complications.

Interleukin-2-dependent augmentation of the anti-TNP antibody production by sodium butyrate in cultured murine splenic B cells.

Previously we found that sodium butyrate (NaBu) markedly enhanced production of the antibody specific for a T-cell-dependent antigen, sheep red blood cells (SRBC) in murine splenocytes (Kishiro, Y., Ueda, K., Fujiwara, M. and Yamamoto, I., Jpn J. Phamacol., 1994 66, 369-376. To gain a better understanding of the target cells for NaBu's action on antibody responses, we have utilized the T-cell-independent antigen, trinitrophenyl-lypopolysaccharide (TNP-LPS) as a stimulant and have examined an effect of NaBu on the anti-TNP antibody production in vitro. NaBu markedly increased the anti-TNP plaque-forming cell (PFC) responses in murine whole splenocytes, but not in murine splenic B cells. Addition of T-cells or the concanavalin A supernatant (CAS) from murine splenocytes to the B cell cultures completely restored the enhancing effect of NaBu. This effect of CAS was totally blocked by an anti-interleukin (IL)-2 antibody and partially by an anti-IL-1 beta or anti-IL-4 antibody. The full enhancing effect of NaBu was also detected when IL-2 was added to the B cell cultures, while IL-2 alone had no stimulatory effect on the control PFC response. IL-1 beta alone significantly stimulated the antibody production and adding NaBu to this IL-1 beta-supplemented culture caused a further increase. Neither IL-4 alone nor NaBu plus IL-4 had any effect on the PFC response. NaBu did not affect the expression of the IL-2 receptor alpha- and beta-chains in B cells stimulated with TNP-LPS. These results suggest that NaBu is an agent that promotes B cell differentiation in vitro in an IL-2-dependent manner.

Intrapancreatic accessory spleen diagnosed by technetium-99m heat-damaged red blood cell SPECT.

This is a report of accessory spleen located within the tail of the pancreas which mimicked a tumor. The correct diagnosis was made noninvasively with 99mTc-HDRBC (heat-damaged red blood cell) SPECT.

[Efficiency of segmental SMANCS/Lip-TAE for hepatocellular carcinoma--comparative studies in the efficacy of segmental SMANCS/Lip-TAI].

We compared the segmental SMANCS/Lip-TAI and the segmental SMANCS/Lip-TAE and studied the effectiveness of both treatments and the influence and/or the side effects on liver function. In resected cases, we studied histopathologic examination. The response rate of the group treated by TAI was 28.6%, and that of the group treated by TAE was 76.5%. In the group treated by TAE, the therapeutic effects were good in nodular type HCC, using small doses of SMANCS. In both groups, the incidence and degree of side effects showed no significant difference. Hepatic insufficiency occurred in a few cases of the group treated by TAI. In resected cases, viable areas remained below the tumor capsule. In conclusion, segmental SMANCS/Lip-TAE seemed to be an effective treatment without any serious complications.

Newly developed transurethral radiofrequency thermotherapy device for benign prostatic hyperplasia: a pilot study in canine prostate.

The safety and efficacy of transurethral microwave thermo-therapy (TUMT) for the treatment of symptomatic benign prostatic hyperplasia (BPH) have been demonstrated in the urological literature. Since a radiofrequency electromagnetic wave has deeper and more even transmission of heat to tissues than a microwave, we have developed a new prototype device for transurethral thermotherapy using a radiofrequency electromagnetic wave. The device consists of a microcomputer-controlled heat generator operating at 8 Mhz, a temperature monitoring system, a urethral cooling system and a urethral applicator and a rectal thermosensor probe. A balloon electrode encased in a specialized Foley catheter is connected parallel to twin plate electrodes on both sides of the pelvic region. An 8-MHz electromagnetic wave is directed to the prostate by means of capacitive coupling. The water coolant continuously perfused through the catheter allows high temperatures within the prostate while preserving the urethral mucosa. Heating experiment using agar phantom showed the hot spots to be distributed at 0.5-3 cm from the catheter surface. Heating experiment using canine prostates demonstrated that an intraprostatic temperature of > 48 degrees C could be achieved while the urethral and rectal temperatures had not exceeded 36 and 40 degrees C respectively. Histological examination immediately after the experiment showed the urethral mucosa to be preserved while coagulation necrosis of the periurethral prostate accompanied with congestion and hemorrhage of small blood vessels were observed at 5-8 mm from the urethra. The bladder and the rectum showed no gross alterations. Histopathological examination 10 days after the experiment revealed the intact urethral mucosa and mild mononuclear infiltration around the destroyed periurethral glands.(ABSTRACT TRUNCATED AT 250 WORDS)

Induction of hepatocyte growth factor in human skin fibroblasts by epidermal growth factor, platelet-derived growth factor and fibroblast growth factor.

Hepatocyte growth factor (HGF) is a potent mitogen for rat and human hepatocytes in primary culture and appears to be the physiological hepatotrophic factor that triggers or modulates liver regeneration. Regulation of HGF gene expression and the protein production in human skin fibroblasts was examined. Addition of epidermal growth factor (EGF), platelet-derived growth factor (PDGF), basic fibroblast growth factor (bFGF), acidic fibroblast growth factor (aFGF) and transforming growth factor-alpha (TGF-alpha) to confluent cultures of the cells markedly stimulated HGF secretion from the cells. The stimulating effect of EGF, PDGF and bFGF was further investigated. The effect of all three growth factors was maximal at 3-30 ng/ml and was accompanied by an increase in HGF mRNA levels. The mRNA levels were not elevated at 5 h but were at 10 h or more after addition of EGF. The levels of HGF mRNA in fibroblasts treated with the optimal doses of EGF, PDGF, bFGF, aFGF and TGF-alpha for 24 h were 6, 4, 5, 4 and 5 times that of control cultures incubated in medium only, respectively. The growth factor-induced HGF mRNA expression and HGF secretion was inhibited by addition of TGF-beta 1 or dexamethasone. Pretreatment with a high dose of phorbol 12-myristate 13-acetate (PMA), which causes down-regulation in protein kinase C (PKC) activity and PMA-induced HGF secretion, did not reduce the effects of the growth factors on HGF mRNA expression and HGF secretion, but rather enhanced them.(ABSTRACT TRUNCATED AT 250 WORDS)

Alendronate modulates osteogenesis of human osteoblastic cells in vitro.

The bisphosphonates, which are carbon-substituted pyrophosphates, have been studied extensively both in vivo and in vitro to elucidate their effects on bone tissues and cells. However, because these agents were shown to have a potent inhibitory effect on bone resorption, the majority of studies have focused on only this aspect of bone metabolism. There appears to be less information regarding the direct effect of bisphosphonates on bone formation, so thus we undertook experiments to investigate the effects of bisphosphonates, especially alendronate, on the mineralization and matrix protein synthesis of human osteoblastic cells in vitro. The data show that the bisphosphonates, alendronate, etidronate and pamidronate, suppressed 1,25-dihydroxycholecalciferol (1,25(OH)2D3)-stimulated mineralization of human osteoblastic cells at high concentrations, while relatively lower concentrations of alendronate and etidronate potentiated mineralization of the cells in the presence of 1,25(OH)2D3. The potentiation of mineralization with alendronate was accompanied by increased synthesis of bone matrix proteins, osteocalcin and collagen, and the mRNA of pro alpha(I) collagen. These findings show that in addition to their well-known effects on bone resorption, bisphosphonates have significant and direct effects on osteogenesis in osteoblasts in vitro. The actual mechanism remains to be further investigated.

Enhancement of in vitro antibody production of murine splenocytes by ascorbic acid 2-O-alpha-glucoside.

Using an antigen-specific plaque forming cell (PFC) assay, we have studied the effect of ascorbic acid 2-O-alpha-glucoside (AA-2G), a new stable derivative of ascorbic acid (AsA), on antibody production in cultured murine splenocytes in comparison with that of AsA. A single addition of AA-2G (0.0625-1.0 mM) remarkably stimulated both anti-SRBC (sheep red blood cell, TD antigen) and anti-TNP (trinitrophenyl, TI antigen) PFC responses in a dose-dependent manner, although AsA failed to stimulate their responses. However, repeated additions of AsA at 12-h intervals during the cultivation resulted in enhancement of the anti-SRBC PFC response, and the magnitude of stimulation was comparable to that obtained by a single addition of AA-2G. AA-2G's effect was abrogated in the presence of castanospermine (alpha-glucosidase inhibitor) in the medium, indicating that the immunostimulation brought about by AA-2G is attributed to AsA released from the glucoside by alpha-glucosidase of cultured cells. In fact, the cells maintained a certain amount of AsA for a relatively long time after exposure to AA-2G. In contrast, AsA levels in the cells treated with AsA quickly decreased. AA-2G markedly enhanced lipopolysaccharide(LPS)-induced proliferative response, and could affect the concanavalin A (Con A) response weakly. These results suggest that AsA effectively potentiates B-cell functions in the humoral immune system. Thus, we conclude that AA-2G is capable of enhancing antibody production in the cultured splenocytes via a continuous supplementation of AsA and that AA-2G is available for long-term cell cultures as a useful tool for analyzing biological function of AsA.

Suppression of 125I-uptake in mouse thyroid by seaweed feeding: possible preventative effect of dietary seaweed on internal radiation injury of the thyroid by radioactive iodine.

We conducted an animal experiment to determine how dietary seaweeds rich in iodine and dietary fibers suppress radioactive iodine uptake by the thyroid, using mice and four kinds of experimental diets, three with 1% or 2% powdered fronds of the kelp Laminaria religiosa and 2% powdered laver Porphyra yezoensis, and one with cellulose. Iodine content of a hot-water extract of the kelp was 0.530 +/- 0.001%, and its dietary fiber (DF) values were 52.8 +/- 1.2%. Iodine in an extract of the laver was 0.008 +/- 0.001%, and its DF values were 41.4% +/- 0.7%. A statistically significant reduction of 125I uptake by the thyroid, 3 hours after intragastric administration of the radionuclide at a dosage of 18.5 kBq or 185 kBq in 0.3 ml aqueous solution per mouse, was observed in mice previously fed the experimental diets containing 1% and 2% kelp during periods varying from 24 hours to 7 days. The degree of the suppression was observed to depend on the amount of iodine in the diet or in the injected sample, no matter whether organic or inorganic, judging from the results of an additional experiment. Thus, we conclude that previously fed iodine-rich material, especially dietary seaweeds rich in iodine and other minerals, vitamins, and beta-carotene, such as kelps or laver supplemented with inorganic iodine, may be effective in prevention of internal radiation injury of the thyroid.

[Phosphorus 31-magnetic resonance spectroscopic studies of animal liver after extracorporeal shock wave lithotripsy].

The technique of magnetic resonance spectroscopy has been developed, and the study of high-energy phosphate metabolites in the liver using phosphorus 31 magnetic resonance spectroscopy (31P-MRS) has been reported in humans and animals, but few studies have used 31P-MRS for the evaluation of extracorporeal shock wave lithotripsy (ESWL) of the liver. In this study, 31P-MRS was used to evaluate the metabolic changes in hamster liver after ESWL and histological correlation was performed. Syrian golden hamsters were anesthetized and shock waves were irradiated to the left side of the liver. Hamsters were irradiated by LITHOSTAR-PLUS (SIEMENS) at a voltage of 19 KV. 31P-MRS was studied by JNM-GSX model 270 (6.34 Tesla). Typical peaks of 31P-spectra of hamster liver showed a tendency for PDE/beta-ATP, alpha-ATP/beta-ATP and gamma-ATP/beta-ATP to decrease among the irradiated group compared with the control group. However, there were no significant differences in PME/Pi, beta-ATP/Pi or (alpha-ATP-beta-ATP)/beta-ATP between the control group and irradiated group. With regard to intracellular pH and PDE/beta-ATP, a decreasing tendency was noted in the irradiated groups (p less than 0.05). There was no difference in the signal intensity of T1WI and T2WI on 1H-MRI, between these two groups. Pathologically, the irradiated group showed minor hemorrhage and edema in the liver, and subcapsular hematoma. The results obtained from 31P-MRS clearly showed the metabolic changes and were correlated well with the histological findings, but MRI was not capable of providing close visualization of post-ESWL liver damage.(ABSTRACT TRUNCATED AT 250 WORDS)