Effective application of ET-Kyoto solution for clinical lung transplantation.

The shortage of lung donors and ischemia-reperfusion injury following transplantation have been grave problems in lung transplantation (LTx). One of the most important strategies to solve these problems is the development of effective and highly reliable methods for lung preservation. Therefore, we developed a new organ preservation solution, namely, the extracellular-type trehalose-containing Kyoto (ET-Kyoto) solution. Here we report the first experience of clinical application of ET-Kyoto solution for cadaveric LTx. The recipient was a 38-year-old man with pulmonary emphysema. The donor was a 51-year-old male current smoker with a smoking history of 62 pack-years. The ventilated donor's PaO(2) was 340 Torr (FiO(2) = 1.0). The pulmonary vasculature was flushed with ET-Kyoto solution supplemented with nitroglycerine and dibutyryl cAMP. The recipient underwent bilateral sequential LTx on cardiopulmonary bypass. The ischemic time was 544 and 613 minutes for the left and right lung, respectively. PaO(2) (FiO(2) = 1.0) was 385 Torr immediately after reperfusion. The donor lung was so large that bilateral partial resections were performed at 413 minutes (right) and 348 minutes (left) after reperfusion. On histopathologic examination of the resected transplanted lungs the structure was almost normal. Postoperatively, PaO(2) (FiO(2) = 1.0) was over 400 Torr with or maximum of 526 Torr. The clinical course was almost uneventful. In conclusion, ET-Kyoto solution may be safely applied in clinical cadaveric LTx with extended donor lungs and relatively long ischemic times. Functional and histopathological efficiency of ET-Kyoto solution was confirmed. Longer preservation times with preserved quality using ET-Kyoto solution would increase the donor pool and enable semielective LTx.

Prophylactic chemolipiodolization for postoperative hepatoma patients.

BACKGROUND/AIMS: The efficacy of prophylactic chemolipiodolization following hepatic resection in patients with hepatocellular carcinoma was studied. METHODOLOGY: Forty-four of 67 consecutive patients with hepatocellular carcinoma who underwent hepatectomy between 1980 and 1997 were divided into two groups: group A (n = 21), in which prophylactic chemolipiodolization was performed during postoperative follow-up (2.4 times on average using a 39 mg mean dose of epirubicin or doxorubicin); and group B (n = 23), without prophylactic chemolipiodolization. The clinicopathological background and patient survival were compared retrospectively. RESULTS: There were no differences in the clinicopathological background between the two groups. Multiple intrahepatic recurrence was frequently observed in group B (P < 0.02). The recurrence-free survival rates in group A (54.4% and 31.1% at 3 and 5 years, respectively) were significantly higher than those in group B (15.7% and 7.9%, respectively). The survival rates of group A (95.2% and 80.4% at 3 and 5 years, respectively) were significantly higher than those in group B (40.1% and 22.9%, respectively). CONCLUSIONS: Our data suggest that postoperative prophylactic chemolipiodolization can be an effective treatment in reducing intrahepatic recurrence and may prolong survival for hepatocellular carcinoma patients following hepatic resection.

Significance of cyclic adenosine monophosphate and nitroglycerin in ET-Kyoto solution for lung preservation.

BACKGROUND: We previously demonstrated that the supplement of both dibutyryl cyclic adenosine monophosphate (db-cAMP) and nitroglycerin to the conventional ET-Kyoto solution improved lung preservation significantly. However, the significance of each component in lung preservation remained unclear. We examined the efficacy of the two components on lung preservation in the current study. METHODS: Rat lung grafts (eight per group) were studied in an isolated lung perfusion model. Group 1 grafts were flushed and preserved with ET-Kyoto solution containing 2 mmol/L of db-cAMP. Group 2 grafts were flushed and preserved with ET-Kyoto solution containing 100 mg/L of nitroglycerin. In group 3, the grafts were flushed and preserved with ET-Kyoto solution containing neither db-cAMP nor nitroglycerin as control group. After 4-hour cold storage, the lung grafts were reperfused for 50 minutes. RESULTS: The lung grafts in groups 1 and 2 showed significantly better lung function after reperfusion than those in group 3 with regard to arterial oxygen tension, shunt fraction, peak inspiratory airway pressure, mean pulmonary arterial pressure, and pulmonary vascular resistance. The supplementation of db-cAMP improved especially the pulmonary arterial pressure and pulmonary vascular resistance, while the supplementation of nitroglycerin improved especially the oxygenation and airway pressure of the grafts. CONCLUSIONS: Both of db-cAMP and nitroglycerin had beneficial effects on lung preservation and are essential to the ET-Kyoto solution. There was a difference between the two components in the effects on preserved lungs.

Effective 30-hour preservation of canine lungs with modified ET-Kyoto solution.

BACKGROUND: With the aim of developing a preservation solution that can preserve donor lungs reliably for a long time, we prepared a modified ET-Kyoto solution by adding N-acetylcysteine, nitroglycerin, and dibutyryl adenosine 3', 5'-cyclic phosphate to the previously reported ET-Kyoto solution, which contains trehalose, gluconate, and hydroxyethyl starch. In this study, we examined the efficacy of modified ET-Kyoto solution in 30-hour lung preservation. METHODS: Twenty five pairs of adult mongrel dogs were divided into four groups. Donor lungs were flushed with modified ET-Kyoto solution (n = 9), with ET-Kyoto solution (n = 6), with University of Wisconsin solution group (n = 6), or with ET-Kyoto solution plus the solvents of nitroglycerin (ethanol and propylene glycol) (n = 4), then stored at 4 degrees C for 30 hours. All animals were treated with prostaglandin E1. Left lungs were transplanted and reperfused for 6 hours. RESULTS: With respect to arterial oxygen tension, peak inspiratory pressure, and wet-to-dry lung weight ratio, modified ET-Kyoto solution was significantly superior to ET-Kyoto solution. The modified ET-Kyoto solution was significantly superior to University of Wisconsin solution with respect to survival rate, arterial oxygen tension, and wet-to-dry lung weight ratio. Ultrastructural findings supported these results. CONCLUSIONS: These results suggest that modified ET-Kyoto solution is superior to University of Wisconsin solution for 30-hour lung preservation.

Dissociation between the mitogenic effect and antitumor activity of seed extract from Aeginetia indica L.

The extract from seeds of Aeginetia indica L. (AIL), a parasitic plant, induces potent antitumor immunity against Meth A fibrosarcomas in BALB/c mice. AIL also possesses a thymocyte co-stimulatory effect in vitro with suboptimal dose of Con A, a B cell mitogenic effect, and stimulates AIL-primed CD4+ T cells to produce Th1-type cytokines. In this study, we investigated the relationship between mitogenicity and antitumor activity with AIL. When AIL was analyzed by SDS-PAGE, there was strong and diffuse staining in the region between 14 kDa and the bottom of polyacrylamide gel and it was unaffected when AIL was digested with proteinase K (PK) before SDS-PAGE. Some bands with different molecular mass were also found in silver-stained gel and they disappeared completely by incubating AIL with PK before SDS-PAGE. The in vitro thymocyte co-stimulatory and B cell mitogenic effects were not influenced by digesting AIL with PK but were completely suppressed by the oxidation of AIL with sodium periodate before culture. In contrast, the in vivo antitumor activity was completely abolished by PK, but it was not affected by periodate oxidation. We generated mAbs specific for AIL and investigated the influence on the antitumor activity of AIL in vivo. Around 60-80% of tumor-bearing mice failed to recover from a challenge tumor when they were treated with supernatants isolated from mAb-induced precipitation reactions. Immunoblotting (Western blotting) revealed that all the mAbs reacted exclusively with a 50-60 kDa protein and that this reactivity was not influenced after oxidizing the blots with sodium periodate. We demonstrated that AIL contains polysaccharides and proteins. The polysaccharides induced B cell mitogenic and thymocyte co-stimulatory effects in vitro, while the proteins, especially a 50-60 kDa protein containing non-carbohydrate epitopes recognized by the mAbs, mediated antitumor activity in vivo.

Prostanoids and cough response to capsaicin in asthma and chronic bronchitis.

Cyclooxygenase products are released by chronic airway inflammation. Our working hypothesis for the present study was that prostanoids augment airway cough sensitivity. The effects of a cyclooxygenase inhibitor, indomethacin (100 mg.day-1 for 4 days), and a thromboxane synthesis inhibitor, OKY-046 (400 mg.day-1 for 4 days), on cough response to inhaled capsaicin were examined in eight patients with asthma, 10 patients with chronic bronchitis, and 10 normal subjects. Capsaicin cough threshold, the lowest concentration of capsaicin eliciting five or more coughs, was measured as an index of airway cough sensitivity. In asthmatics, the cough thresholds with indomethacin treatment (15.7 (GSEM 1.38) microM) and OKY-046 (10.2 (GSEM 1.20) microM) were significantly greater than the value with placebo (6.05 (GSEM 1.25) microM). In patients with chronic bronchitis, the cough threshold was significantly greater with indomethacin (5.94 (GSEM 1.50) microM) than with placebo (3.41 (GSEM 1.33) microM and OKY-046 2.97 (GSEM 1.43) microM). In normal subjects, the capsaicin cough threshold was not altered by indomethacin or OKY-046 treatment. These results support our hypothesis and suggest that thromboxane A2 may be one of the cyclooxygenase products augmenting airway cough sensitivity in asthma, but not in chronic bronchitis.

Characteristics of antitumor activity of 3,4-dihydro-6-[4-(3,4-dimethoxybenzoyl)-1-piperazinyl]- 2(1H)-quinolinone (vesnarinone) against a human adenoid squamous carcinoma-forming cell line grown in athymic nude mice.

The tumors produced by transplantation into nude mice of human adenoid squamous carcinoma-forming cell line TYS, presumably derived from a minor salivary gland, were treated with a differentiation-inducing agent, vesnarinone, which was given per o.s. daily at a dose of 200 mg/kg for 35 days. They were then examined morphologically and immunohistochemically. The vesnarinone treatment resulted in a significant suppression of tumor growth. In addition, tumor nests indicating keratinocyte and acinar cell differentiation were often observed in the treated tumors, but not in untreated controls. Tissue sections from vesnarinone-treated and untreated TYS tumors were stained with monoclonal antibody (NAb) directed to carbohydrate antigen LeY or proliferating cell nuclear antigen (PCNA) and with rabbit polyclonal antibody to p53. Antibody staining patterns were compared with morphological characteristics of cells as revealed by hematoxylin and eosin staining, and DNA fragmentation patterns as revealed by 3'-OH nick-end labelling techniques. Tissue sections from vesnarinone-treated TYS tumors showed positive reaction with nick-end labelling and were extensively stained strongly by anti-LeY MAb, whereas the untreated tumors showed negative reaction with nick-end labelling and were infrequently stained by anti-LeY MAb. Within LeY-positive areas of tissue sections from the vesnarinone-treated tumors, keratinocyte and acinar cell differentiation as well as DNA fragmentation were frequently observed, although not all LeY-positive cells showed such signs of apoptosis. LeY-positive cells showed consistent negative staining by anti-PCNA MAb and anti-p53 rabbit serum. From these findings, it can be considered that vesnarinone has differentiation and apoptosis-inducing activity against TYS cells grown in athymic nude mouse.

Seed extract of Aeginetia indica L induces cytokine production and lymphocyte proliferation in vitro.

We previously reported that the extract of seeds from Aeginetia Indica L (AIL), a parasitic plant, induces potent antitumor immunity in tumor-bearing mice and that CD4+ T cells appear to be the main contributors in the induction of antitumor resistance. The present study was set up to investigate the in vitro effects of AIL on various lymphoid cells. Spleen cells from mice pretreated with AIL every 2 days for 1 week produced interleukin 2 (IL-2), interferon gamma (IFN gamma), tumor necrosis factor (TNF) and interleukin 6 (IL-6) when these cells were stimulated in vitro by AIL. Further, we found that CD4+ T cells were main producers of IL-2 and TNF upon the stimulation with ALL in vitro, while both CD4+ and CD8+ T cells secreted IFN. On the other hand, ALL was mitogenic in vitro to T enriched splenic lymphocytes as well as B enriched splenic lymphocytes. Moreover, AIL also proliferated thymocytes and this activity was potently synergistic with a suboptimal dose of concanavalin A (Con A). Lipopolysaccharide (LPS) contamination in AIL preparation was negligible since proliferative activity of AIL to B enriched splenic lymphocytes was not influenced in the presence of an endotoxin antagonist, polymyxin B sulfate (PMB). Further, B cell mitogenic activity of AIL seems to be mediated by different mechanism(s) from that of LPS since ALL could proliferate B enriched lymphocytes of C3H/HeJ mice which do not respond to the stimulation with LPS. A well known biological response modifier (BRM), Krestin (PSK), had no ability in inducing either T or B lymphocyte activation in vitro as shown by AIL.(ABSTRACT TRUNCATED AT 250 WORDS)

[A case of pneumonitis and hepatic injury caused by a herbal drug (sho-saiko-to)].

We report a case of pneumonitis and hepatic injury caused by Sho-saiko-to. A 56-year-old man was admitted to our hospital because of hepatic disorder. The levels of serum transaminases returned to normal within two months without specific treatment and he was discharged. Four weeks later, he was readmitted because of severe pneumonitis and mild hepatic disorder. Under the suspicion of drug-induced pneumonitis, all medications were discontinued and high-dose glucocorticoid including "pulse therapy" was given. Consequently, pneumonitis and hepatic function markedly improved. Careful history taking revealed the ingestion of Sho-saiko-to before both admissions. Lymphocyte stimulation test against Sho-saiko-to was positive. Challenge test using Sho-saiko-to resulted in decrease of PaO2 and elevation of serum transaminases. Based on these findings, the diagnosis of pneumonitis and hepatic injury induced by Sho-saiko-to was established.

An extract of seeds from Aeginetia indica L., a parasitic plant, induces potent antigen-specific antitumor immunity in Meth A-bearing BALB/c mice.

The antitumor activity of an extract of seeds from Aeginetia indica L., a parasitic plant, was investigated. BALB/c mice, inoculated i.p. 1 x 10(5) syngeneic Meth A tumor cells, were administered 2.5 mg/kg A. indica extract i.p. every 2 days from day 0. The untreated mice died of an ascitic form of tumor growth within 21 days, whereas all the treated mice completely recovered from tumor challenge without any side-effects. The extract did not exert direct cytotoxic activity against Meth A in vitro. Mice that survived after the first challenge as a result of A. indica treatment overcame the rechallenge with homologous Meth A without additional administration of the extract. On the other hand, those mice could not survive after rechallenge with Meth 1 tumor cells, which were also established in BALB/c mice but were different in antigenicity from Meth A, suggesting the development of antigen-specific concomitant immunity in the A. indica-cured mice. In the induction phase of antitumor resistance in this system, CD4+ T cells appeared to be the main contributors, since in vivo administration of anti-CD4 mAb completely abolished such resistance. In contrast, anti-CD8 mAb administration did not influence the effect of A. indica. The importance of CD4+ T cells in antitumor immunity was again clarified by Winn assay; that is, spleen and lymph node cells depleted of CD4+ T cells in vitro prior to assay abolished antitumor activity on co-grafted Meth A tumor cells in vivo.

[A case report of carcinoid of the rectum coexisting with adenocarcinoma in a single lesion].

A 69-year-old female, chiefly complaining of diarrhea, was admitted to the Toyama Medical & Pharmaceutical University Hospital. A barium enema and an endoscopic examination revealed a villous adenocarcinoma of the rectum. Computerized tomography disclosed multiple metastatic lesions in the liver. She underwent an amptatio recti and an intra-arterial cannulation of the common hepatic artery in order to administer anticancer agents. Gross findings of the resected specimen showed villous adenocarcinoma containing a small ulcerated lesion. Histologic findings of this lesion revealed a collision neoplasm containing both an adenocarcinoma and a carcinoid with no obvious transitional region between the two lesions. The type D carcinoid, determined according to Soga's histologic classification, showed positive argyrophil granules but no argentaffin granules in the cytoplasm. The electron-microscopic findings of the carcinoid revealed small electron-dense endocrine granules measuring about 150 approximately 300 nm in diameter. The woman died 4 months after this operation.

Monoamine fluorescence in the median eminence of the Japanese quail, Coturnix coturnix japonica, following medial basal hypothalamic deafferentation.

Monoamine fluorescence was examined in the ventral hypothalamus of the Japanese quail, Coturnix coturnix japonica after medial basal hypothalamic deafferentation. In sham-operated control birds, numerous yellow-green fluorescent fibers were observed in the median eminence and the nucleus tuberis. In the area of the paraventricular organ, a number of fluorescent fibers and cell bodies were observed. In birds with deafferented hypothalami, fluorescence disappeared both in the median eminence and the nucleus tuberis. In the area of the paraventricular organ, which was within the area of deafferentation, fluorescence of neuronal perikarya did not change, but fluorescent fibers decreased markedly in number. Disappearance of monoamine fluorescence in the median eminence and the nucleus tuberis is discussed in relation to the tanycyte absorptive function and gonadaly development.