Formation and inhibition of genotoxic malonaldehyde from DNA oxidation controlled with EDTA.

The attack of DNA by reactive oxygen species, such as hydroxyl radical, causes several types of damage, which subsequently promote diseases. Determination of oxidized products, such as malonaldehyde (MA), from DNA would provide theoretical and practical information on the mechanisms of DNA oxidation following DNA damage and this information could be used to prevent DNA damage caused by oxidation. In the present study, calf thymus DNA was oxidized by Fenton's reagent/EDTA with or without natural antioxidants-flavonoids and anthocyanins-and synthetic antioxidants, Trolox and 2H-pyrrole, 3,4-dihydro-2,2-dimethyl-, 1-oxide (DMPO). Amounts of MA formed, which was determined by gas chromatography, in oxidized DNA with the presence of antioxidants, ranged from 7.35+/-0.88 nmol/mg (2''-O-GIV) to 12.6+/-0.24 nmol/g (cyanidin). Except for cyanidin, all antioxidants tested inhibited MA formation. DMPO and Trolox inhibited MA formation by 12.4% and 27.3%, respectively from oxidized DNA. The decreasing order of inhibitory effect by the anthocyanins was callistephin (30.2%)>keracyanin (27.3%)>Pelargonindin (10.1%)>cyanidin (0%). The decreasing order of inhibitory effect by the flavonoids was 2''-O-GIV (42.7%)>catechin (8.8%)>quercetin (36.4%)>apigenin (34.4%). It is hypothesized that EDTA controlled formation of hydroxyl radicals via trapping Fe(II) ions reversibly.

Enhancement of esophageal carcinogenesis induced in rats by N-amyl-N-methylnitrosamine in the presence of 12-O-tetradecanoylphorbol-13-acetate.

The effects of 12-O-tetradecanoylphorbol-13-acetate (TPA), a tumor-promoting phorbol diester, were determined with regard to the induction of esophageal cancer in Wistar rats following a low-dose initial administration of the esophageal carcinogen N-amyl-N-methylnitrosamine [(AMN) CAS: 13256-07-0]. The induction of esophageal cancer was enhanced by TPA given in drinking water after AMN administration; i.e., the incidence of developing esophageal cancers and the multiplicity (number of esophageal cancers per rat) were significantly higher in groups given TPA solution orally after an oral administration of AMN than in those given the AMN solution alone. The enhancement of carcinogenesis with TPA was not affected by the interval between the administration of AMN and the administration of TPA. However, pretreatment with TPA before AMN administration did not enhance the induction of esophageal cancer. Neoplasms were not detected in groups given only TPA or tap water. Because this approach is similar to the phenomenon of two-stage carcinogenesis in the skin, it should provide a meaningful experimental model for studying two-stage carcinogenesis in the esophagus.

Chemical removal and recovery of phosphorus from excess sludge in a sewage treatment plant.

We describe a process for the recovery of phosphorus from excess sludge in a sewage treatment plant that currently uses polyaluminium chloride for chemical phosphorus removal. Instead, we employed alkaline dissolution of excess sludge with calcium phosphate precipitation to recover phosphorus from sewage. The recovery ratio for phosphorus from sewage using the phosphorus recovery system is approximately 50%. In addition, the amount of excess sludge in the phosphorus recovery system is approximately half that of conventional chemical phosphorus removal. Alkaline dissolution of excess sludge resulted in dissolution of aluminium into the supernatant. Furthermore, since dissolved aluminium can be reused as a coagulant, the phosphorus recovery system could be used to economize coagulant consumption. Operation and maintenance costs of the phosphorus recovery system are 25.9 U.S. cents per 1 m3 of sewage compared to 32.0 U.S. cents per 1 m3 of sewage for conventional chemical phosphorus removal, representing a decrease of 20% in the operation and maintenance costs.

Heat, drugs, and radiation given in combination is palliative for unresectable esophageal cancer.

From April 1966 to April 1986, 101 men and women with unresectable squamous cell carcinoma of the esophagus were treated in our clinic. Since 1983, 21 were treated with a combination of hyperthermia, chemotherapy, and radiotherapy (group I). Before 1983, for another 80 patients, radiation plus chemotherapy had been prescribed (group II). Nine of 21 patients in group I had an unresectable carcinoma due to an advanced tumor, 9 had an associated severe clinical status, and 3 refused surgery. Out of 80 in group II, 50 had a far advanced tumor, 21 had a poor clinical condition, and 9 refused operation. With regard to staging, for 21 in group I, 6 were classified as Stage I, 5 as Stage II, 7 as Stage III, and 3 as Stage IV. As to the 80 in group II, those in Stage I, II, III, and IV accounted for 8, 22, 39, and 11, respectively. The median doses of each modality, for patients in group I, were 6 times of hyperthermia at 42-45 degrees C for 30 minutes, 40 Gy of X ray and 30 mg of bleomycin. For patients in group II, a median dose of 56 Gy of X ray was given. Response rates determined by esophagograms and endoscopies for the patients in groups I and II were 76.2% (16/21; 4 CRs, 12 PRs) and 39.2% (31/79; 2 Crs, 29 PRs), respectively (p less than 0.001). The effective rates determined by improvement in quality of life (relief of pain and dysphagia) for groups I and II were 61.9% and 37.2%, respectively. A longer survival was obtained for patients in group I (median survival: 9 months vs 6 months). Especially for the patients classified as Stage I, a significantly longer survival was obtained with a combination of hyperthermia, chemotherapy, and radiotherapy (p less than 0.01).

Pre-operative estimation of complete resection for patients with oesophageal carcinoma.

Three hundred and seventy-nine patients were studied retrospectively regarding the possibility of a complete resection of the oesophageal carcinoma based on the combined findings of pre-operative oesophagogoraphy and computed tomography (CT). One hundred and four out of 129 patients (96.1%) having lesions which did not demonstrate all three of the aforementioned factors (a lesion shorter than 8 cm, a normal oesophageal axis, and normal contact of the lesion with neighboring organs in CT) underwent a complete resection of the oesophageal lesion. Fifty-three percent of the patients (52/97) with a lesion showing only one of these factors had a complete resection. Whereas, on the other hand, a complete removal of the malignancy was only possible in 22% of the patients with two or all three of the findings. Moreover, as a result of further analysis limited for resected cases, the number of positive factors in these pre-operative findings correlated with the advancement of the surgical stage, which reflected a curability in surgery and a rate of postoperative complications. In order to make adequate plans for the treatment of patients with advanced oesophageal cancer, the finding of (i) the length of lesion, (ii) a deep ulceration and deformity of the oesophageal axis and (iii) any abnormal contact in CT, are considered to be very useful.

Surgical treatment of a patient with idiopathic portal hypertension and hepatic encephalopathy.

Idiopathic portal hypertension is clinically characterized by splenomegaly and portal hypertension. Hepatic encephalopathy is rare in cases with idiopathic portal hypertension. In a 59-year-old man with recurrent hepatic encephalopathy for one year, a large splenorenal shunt was detected in the computed tomography and angiography, and liver biopsy revealed a portal fibrosis consistent with idiopathic portal hypertension. Devascularization of the upper stomach, splenectomy and closure of the splenorenal shunt were done. The patient has had no experience of encephalopathy since the operation even without drug treatment. Surgery should be considered for treatment of chronic hepatic encephalopathy in patients with idiopathic portal hypertension and portosystemic shunts.

Structural determination of subsidiary colors in commercial Food Green No. 3 (fast green FCF, FD & C Green No. 3).

HPLC analysis revealed that eight subsidiary colors existed in commercial Food Green No. 3 (fast green FCF, FD & C Green No. 3). Among them, four subsidiary colors C, F, G, and H were isolated by using preparative HPLC and their structures were determined by MS and NMR. They were the disodium salt of 2-[[4-[N-ethyl-N-(3- sulfophenylmethyl)amino]phenyl][4-[N-ethyl-N-(4- sulfophenylmethyl)amino]phenyl]methylio]-4-hydroxybenzenesulfonic acid (abbreviated as m,p-G-3), the sodium salt of 2-[[(4-N-ethylamino)phenyl][4-[N-ethyl-N-(3- sulfophenylmethyl)amino]-phenyl]methylio]-4-hydroxybenzenesulfonic acid [abbreviated as HSBA-(EA) (m-EBASA)], the sodium salt of 2-[[(4-N-diethylamino)phenyl][4-[N-ethyl-N-(3- sulfophenylmethyl)amino]phenyl]-methylio]-4-hydroxybenzenesulfonic acid [abbreviated as HSBA-(di-EA) (m-EBASA)], and the sodium salt of 2-[[4-[N-ethyl-N-(phenylmethyl)amino]phenyl][4-[N-ethyl-N-(3- sulfophenylmethyl)-amino]phenyl]methylio]-4-hydroxybenzenesulfonic acid [abbreviated as HSBA-(EBA)(m-EBASA)], respectively. HSBA-(di-EA) (m-EBASA) was a subsidiary color newly found in commercial Food Green No. 3.

[Hyperthermia combined with chemotherapy and irradiation for esophageal carcinoma].

For clinical application in cases of carcinoma of the esophagus, we developed an endotract electrode ("applicator") for radiofrequency (RF) and an RF-generator (13.56 MHz) automatically controlled by a computer monitoring the temperature of the lesion. We then designed a combined therapy of hyperthermia, chemotherapy and radiotherapy (so-called hyperthermo-chemo-radiotherapy: HCR), on the basis of the additive or synergistic effects of hyperthermia combined with radiation and/or anticancer drugs. We applied this combined therapy to patients with esophageal carcinoma, using our newly developed heating system. HCR therapy was preoperatively performed on thirty-three patients with esophageal carcinoma. In the histopathological study of resected specimens, seven out of thirty-three patients (21.2%) showed markedly effective improvement, in seventeen (51.5%) the effects were moderately effective and in nine (27.3%) there were no apparent effects. The effectiveness (markedly + moderately effective/all cases) of preoperative HCR therapy was 72.7% (24/33), and significantly higher than the 50% (54/108) obtained in one hundred and eight patients who were preoperatively prescribed radiotherapy alone or in combination with chemotherapy. We then applied the combined therapy with hyperthermia to sixteen patients with unresectable esophageal carcinoma and eleven of these patients (68.8%) showed partial and moderate responses in esophagography. This combined therapy with hyperthermia is highly effective, and shows great promise for treating patients with esophageal carcinoma.

[Endocrinological and metabolic disorders in the patients with massive bowel resection in the newborn period].

Massive (70%) resection of the small bowel was performed in seven newborn infants. Follow-up study was undertaken in four of these patients and the growth and endocrinological status were evaluated. Body weight and height of three patients were below 50th percentile. On glucose tolerance test, peak IRI was low and insulinogenic indices were below normal range. These findings suggest that there are some mechanisms which suppress the secretion of insulin. Pituitary and thyroid function were within normal limits but secondary sexual manifestation was not seen in a fifteen year old boy. Hyperoxaluria was seen in three children, and one had kidney stone with some impairment of renal function.

Stimulatory action of mitemcinal (GM-611), an acid-resistant non-peptide motilin receptor agonist, on colonic motor activity and defecation: spontaneous and mitemcinal-induced giant migrating contractions during defecation in dogs.

The aim of this study was to characterize giant migrating contractions (GMCs) during spontaneous defecation in dogs and to investigate the effect of mitemcinal (an orally active and highly acid-resistant motilin receptor agonist) on colonic motility to assess the possibility of using it for the treatment of colonic motility disorders. To assess colonic motility, strain-gauge force transducers were implanted on the gastrointestinal tract of five dogs, and the behaviour of the dogs was monitored with a noctovision-video camera system. The effect of mitemcinal (0, 3, 10 or 30 mg per dog) and sennoside (300 mg per dog) on colonic motility was assessed 24 h after oral administration. During a 39-day period, the starting point of most of the 140 GMCs was between the transverse colon and the descending colon, but some variation was observed. In the daytime, the GMCs originated from somewhat more proximal positions than at night. Mitemcinal caused an increase in the GMC-index (integration of contractile amplitude and duration) and proximal translocation of the GMC starting point, but did not cause an increase in the number of defecations 12 h after administration. Sennoside, however, caused a significant increase in the number of defecations, an increase in the GMC-index, and prolongation of the duration of GMCs. The GMC starting point in the canine colon varied during spontaneous defecation. Mitemcinal was a potent prokinetic drug to mimic a spontaneous defecation compared with sennoside. Mitemcinal evacuates more intestinal luminal contents during the defecation than does sennoside.

[Responses of muscle strips from the internal anal sphincter in Hirschsprung's disease to drugs and electrical field stimulation].

Responses of isolated muscle strips from the rat and the dog internal anal sphincter (IAS) to drugs and electrical field stimulation (EFS) were investigated in vitro for the purpose of clarifying a manner of neural control of IAS. Also, responses of muscle strips from IAS of the patients with Hirschsprung's disease were compared with those of muscle strips from human control IAS. Muscle strips from the dog and human IAS as normal control showed contractions to norepinephrine (NE), which were abolished in the presence of phentolamine and relaxations to isoproterenol. EFS (less than 1 msec) induced relaxations of the muscle strips. These responses to EFS were not affected by either one of phentolamine, propranolol and atropine but were inhibited by tetrodotoxin. Muscle strips from IAS in Hirschsprung's disease contracted to both NE and EFS, the responses of which were abolished in the presence of phentolamine. But no relaxation to EFS of muscle strips from IAS in Hirschsprung's disease was observed. These findings revealed that normal IAS is pharmacologically innervated by alpha-adrenergic excitatory nerve, beta-adrenergic inhibitory nerve and non-adrenergic, non-cholinergic inhibitory nerve and suggested that IAS in Hirschsprung's disease is also affected by alpha-adrenergic excitatory nerve but inhibitory neural control is absent.

Determination of endogenous peptides with in vitro ACE inhibitory activity in normotensive human plasma by the fluorometric HPLC method.

An in vitro degradation test of angiotensin (ANG) II or III in normotensive supine human plasma from 9 healthy male subjects confirmed the production of smaller ANG metabolites with angiotensin I-converting enzyme inhibitory activity. These metabolites were identified as ANG (3-8), ANG (5-8), and ANG (3-4), whose respective peptide concentrations were determined by our proposed naphthalene-2,3-dialdehyde (NDA)-HPLC method to be 64 +/- 9, 39 +/- 5, 176 +/- 22, and 197 +/- 35 fmol/ml of plasma.

Combined effects of hyperthermia, bleomycin, and X rays on Ehrlich ascites tumor.

The combined effects of water-bath hyperthermia at 42.5 degrees C for 30 min, 1/10 LD50 Bleomycin iv, and 200 rad x irradiation were studied in DDD strain male mice with Ehrlich ascites tumor. The objective was to acquire data on the optimum regimen for a combined administration of these three modalities. The treatments were given 10 days after the inoculation of 2 X 10(6) of the cells into the right hind limb. Concomitant application of the three modalities led to an 80% regression. A single modality produced no significant effect and a 30-50% regression occurred when only two modalities were combined. To assess the influence of timing and sequence, hyperthermia was applied at 1, 2, 4, and 6 hr before, after, or simultaneously with the combination of Bleomycin and 200 rad X ray. A significant effect was obtained in the case of concomitant application of the three and hyperthermia was effective when applied within 2 hr before or after administration of Bleomycin plus irradiation. This enhancement disappeared at 4-hr intervals.

Two-stage operation for poor-risk patients with carcinoma of the esophagus.

We have devised a two-stage operation for poor-risk patients with carcinoma of the thoracic esophagus. The first-stage operation consists of a right thoracotomy, subtotal esophagectomy, and lymph node dissection. Two to three weeks later, the second-stage operation of esophageal reconstruction with gastric tube is performed under laparotomy. During this 3-week period of no esophagus, the nutritional status can be adequately maintained by intravenous hyperalimentation. We describe herein the technique, postoperative complications, and mortality of our two-stage operation as compared with events during an ordinary one-stage operation for carcinoma of the thoracic esophagus.

Survival rates of women with carcinoma of the esophagus exceed those of men.

Postoperative complications, mortality and survival rates and nuclear deoxyribonucleic acid (DNA) content of cancer cells for 212 male and 47 female patients with carcinoma of the esophagus were studied. There were no differences in age, site and length of carcinoma, histologic type, invasion to the adventitia, metastases to the lymph node, staging and postoperative complications. However, survival time for female patients was significantly longer than in males. In studies of cell nuclear DNA content, the incidence of more favorable DNA type (relatively regular in DNA distribution) was relatively high while the incidence of the unfavorable DNA type (widely scattered DNA distribution) was relatively low in female patients when compared with male patients. These findings suggest that tumor factors warrant attention when studying the clinical differences in males and females with carcinoma of the esophagus.

Radiation-induced esophageal carcinoma responded well to hyperthermic chemotherapy--a case report.

A 66-year-old man with multiple malignant lesions of the esophagus, as apparently induced by irradiation of 60 Gy of 60Co for a mediastinal tumor at the age of 36 y.o., was a poor operative risk because of severe lung disease. This man was treated with a combination of hyperthermia, oral administration of oil Bleomycin-polyacrylate paste and intravenous infusion of cis-Platinum. During 20 administrations of hyperthermia conducted at 42-45 degrees C for 30 min, 600 mg of oil Bleomycin and 200 mg of cis-Platinum, no complications occurred and the heat treatments were completed with no side effects. Repeatedly performed esophagogram and endoscopy showed a complete disappearance of three of the four lesions and a marked regression in the other one. There has been no regrowth or distant spread of the carcinoma up to the present 10 months of follow up. This combination of hyperthermia and chemotherapy may prove to be the optimal strategy for treatment of unresectable esophageal cancer, particularly when radiotherapy is contraindicated.