Successful ex vivo normothermic liver perfusion with purely artificial products using artificial blood.

We tried to make an ex vivo functioning liver with an artificial perfusate that consisted of artificial blood in the pig liver. A liver graft from a female pig weighing 20 kg was harvested in the usual manner. The perfusion solution consisted of artificial blood, L-15 medium, distilled water, bovine serum albumin, NaHCO3, NaOH, KCl, human regular insulin, 50% glucose solution, and dexamethasone. The isolated liver was perfused with this oxygenated perfusate through the portal vein at a rate of 300 ml/min for 9 hours. Seven livers were perfused for 9 hours in this system. Five of the livers showed mean oxygen consumption of over 8 ml-O2/min during perfusion. Histological findings showed that the hepatic architecture was almost completely preserved and numerous hepatocytes exhibited PAS-positive cytoplasmic glycogen deposits in these livers. These observations indicate that we have succeeded in developing an ex vivo functioning liver with an artificial perfusate employing artificial blood.

Experimental study using heparin-immobilized adsorbent of EDA(+)fibronectin.

EDA(+)fibronectin, which might participate in the pathogenesis and/or progress of immune diseases, is efficiently removed from plasma by cryofiltration; however, cryofiltration removes not only EDA(+)fibronectin, but also other proteins. We thus developed a new adsorbent by using its high affinity with heparin. The purpose of this study was to evaluate the efficacy of the adsorbent of EDA(+)fibronectin (OHC-20) in experimental arthritis. The experimental arthritis was induced by injection of 0.5 mg of Mycobacterium butyricum in Lewis rats. Rats were divided into 4 groups; 1 nontreatment group, and 3 treatment groups. Adsorption therapy in treatment groups was performed three times: on Days 1, 3, and 5 in Group A; Days 7, 9, and 11 in Group B; and Days 13, 15, and 17 in Group C. The walking postures of rats improved from dragging to walking on tiptoe, and the increase of hind-foot volume was suppressed in Groups B and C. We conclude that heparin-immobilized adsorbent might be promising for immune diseases.

A chylous cyst of the mesentery: report of a case.

A case is presented of an adult chylous cyst of the mesentery that was preoperatively diagnosed to be a pancreatic cystadenoma. A 66-year-old asymptomatic male was followed up for 15 months under the diagnosis of a benign pancreatic cyst. On October 1997, computed tomography showed a 45 x 40 mm cystic mass in the upper abdomen which came in contact with the pancreas. Endoscopic ultrasonography revealed a multilocular mass with a 7 x 4 mm elevated lesion. Endoscopic retrograde cholangiopancreatography and magnetic resonance cholangiopancreatography revealed the cystic mass to be unrelated to the pancreatic duct. The preoperative diagnosis was a pancreatic cystadenoma or cystadenocarcinoma. A laparotomy showed a 50 x 40 mm cystic mass containing chylous fluid, that arose from the mesentery of the upper part of the jejunum. The pathological diagnosis was a chylous cyst of the mesentery. The preoperative diagnosis in this case was very difficult because the chylous cyst appeared to be attached to the pancreas and this phenomenon is considered to be extremely rare.

New technique of leukocytapheresis by the use of nonwoven polyester fiber filter for inflammatory bowel disease.

Leukocytapheresis (LCAP) is widely used for the treatment of immunological diseases. We studied a new treatment of LCAP using a nonwoven polyester fiber filter. In a basic study, 30-70% of leukocytes were removed. Also, 30-68% of the leukocyte subsets were removed. Sixteen inflammatory bowel disease (IBD) patients, mainly with ulcerative colitis (UC), were treated by this method. Their cytokine activity was normalized in the filter and in the peripheral blood. Eleven of 12 patients with UC were induced to remission. Four patients with Crohn's disease (CD) exhibited improvement. The LCAP using a nonwoven polyester fiber filter was very efficient for treating the patients with IBD. Also, it will be a very useful treatment for immunological diseases and extracorporeal immunomodulation.

Changes of plasma fibronectin in patients treated with cryofiltration for an extended period.

The purpose of this study was to evaluate correlations between extra domain A fibronectin (EDA[+]FN) in plasma and the clinical course in cryofiltration for an extended period. Two patients with systemic lupus erythematosus (SLE), 1 with aortitis syndrome, 1 with ankylosis spondylitis, 1 with polymyositis, 1 with rheumatoid arthritis, and 1 with chronic rejection of a kidney graft, were regularly treated with cryofiltration for more than 3 years. The average level of EDA(+)FN in each year did not show significant change in clinically stable patients with aortitis syndrome, polymyositis, and SLE. In the patient with ankylosis spondylitis, the average level of EDA(+)FN in each year elevated, so cryofiltration was performed frequently. On the other hand, cryofiltration could not attenuate the progression in the juvenile rheumatoid arthritis patient. In the patient with chronic rejection of a kidney graft, kidney function without hemodialysis could be prolonged for 33 months by cryofiltration. The average level of EDA(+)FN elevated as the graft function got worse. Changes of average level of EDA(+)FN in plasma corresponded with changes in the clinical courses of patients with autoimmune disease and chronic rejection of a kidney graft. The EDA(+)FN level might give prognostic information and determine the interval of cryofiltration.

Cryofiltration apheresis for major ABO-incompatible kidney transplantation.

The barrier of ABO-incompatible kidney transplantation is the presence of anti-A and anti-B antibodies in the recipient's circulating blood. Double filtration plasmapheresis (DFPP) is usually used to eliminate those antibodies. We tried cryofiltration apheresis (CRYO) in 2 recipients. Patient 1 was a 45-year-old male with B, Rh(+). The titers of IgM anti-A antibody were only reduced from x64 to x32 by the end of 3 sessions of standard CRYO. Renal allografting was not performed. Case 2 was a 29-year-old male with B, Rh(+). CRYO was introduced for 3 sessions. The initial IgM and IgG titers were x128 and negative, respectively. The standard CRYO system was modified by temperature, treated volume, and filter pore size. The IgM anti-A antibody titer was markedly reduced to x2 after the final session of CRYO. The donor was a 56-year-old father with A, Rh(+). Tacrolimus, azathioprine, methylprednisolone, and antilymphocyte globulin were used as the introductory immunosuppression therapy.

Reduction of EDA(+) fibronectin and its clinical importance on cryofiltration.

Cryofiltration (CRYO) removes cryogel, which is a combination of fibrinogen (Fbg) and fibronectin (FN), containing pathological substances. The purpose of this study was to measure cryogel EDA(+) FN and study the relationship between EDA(+) FN and clinical symptoms in patients with rheumatoid arthritis, SLE and polymyositis. Cryogel contains 51 times more EDA(+) FN than plasma. The patients with rheumatoid arthritis showed a high level of EDA(+) FN in their plasma, and the EDA(+) FN level in plasma corresponded with changes in joint pain. We calculated the clearance level at several points in cryofiltration, and the reduction enabled us to evaluate the CRYO device. The EDA(+) FN clearance was 23.3 +/- 6.4 ml/min, the pFN clearance 16.5 +/- 4.1 ml/min, and the Fbg clearance 22.9 +/- 5.7 ml/min. As the plasma flow in cryofiltration was 30 ml/min, a clearance of EDA(+) FN and Fbg, approximately 23 ml/min, was obviously high. The study of the plasma level change of EDA(+) FN during cryofiltration revealed a temporary elevation. These results suggest that the EDA(+) FN was most efficiently reduced by cryofiltration, would become a good indicator on plasmapheresis, and might move from other tissues into the blood during cryofiltration.

Artificial conditioner for stored organs.

We have developed an artificial organ conditioning system in order not only to condition but also evaluate the viability for transplant graft of kidneys which have been stored for a long time and damaged by warm ischaemia following cardiac arrest. The conditioning system consisted of an artificial lung, a roller pump, an organ chamber and perfusate. The perfusate was prepared with electrolytes, fluorocarbon, amino acid, glucose, an oxygen scavenger and so on. Conditioning was performed by continuous perfusion under mild hypothermia at 24 degrees C. Mildly damaged kidneys (0 and 30 minutes warm ischaemia rabbit kidneys) were well conditioned but severely damaged kidneys failed to produce urination. Our device successfully exposed the viability of stored kidneys and the successful conditioning of damaged kidneys due to warm ischaemia avoiding transplantation. By establishing our method, the harvesting of kidneys following cardiac arrest will be feasible.

[An autopsy case of pulmonary metastasis of cholangiocellular carcinoma associated with marked fibrotic change of the lungs].

An autopsy case of pulmonary metastasis of cholangiocellular carcinoma is presented. A 44-year-old woman was admitted to our hospital because of dyspnea, general fatigue and a sense of abdominal fullness on February 5, 1990. In November 1986, at an other hospital, she had been diagnosed as having diffuse metastatic lung tumor and multiple bone metastases, by transbronchial lung biopsy and other examinations. During the clinical course, she was not received chest irradiation and chemotherapy which induced fibrotic change of lungs. Chest X-ray film on December 21, 1986 showed diffuse nodular shadows in both lung fields. Chest X-ray film on February 4, 1990 showed diffuse reticular shadows with marked shrinkage of lung fields. She died two months after admission. The primary site of the carcinoma was not determined clinically, but was revealed by autopsy to be cholangiocellular carcinoma of the liver, with generalized metastasis. Microscopic findings of the autopsied lung showed markedly increased connective tissue around bronchi and blood vessels, in areas where microtubular adenocarcinoma was scattered. This is a very rare case of pulmonary metastasis of cholangiocellular carcinoma, associated with marked fibrotic change of the lungs during about 3.5 years. To our knowledge, this is the first reported case.

[Cytotoxic effect of anticancer drug combined with anti-AFP antibody on human hepatoma cell in vitro].

The cytotoxic effect of the anticancer drug, Daunorubicin, combined with the anti human AFP horse antibody (Conjugate) on AFP-producing cells and non producing cells was studied in vitro. No different cytotoxic effects between the conjugate and the anticancer drug alone were observed on non-AFP-producing cells. On the other hand, the conjugate had a much stronger cytotoxic effect than the anticancer drug alone against AFP-producing cells when it was incubated for 24 hours, 5 x 10(-1) micrograms/ml as an anticancer concentration. The cytotoxicity of the conjugate is much stronger than of the comcrued effect of the anticancer drug and the anti AFP antibody (individually). The amount of AFP antibody used was very small. Therefore, the effect of the anticancer drug plus the small amount of AFP antibody was almost the same as the effect of anticancer drug alone. Under these conditions melting of nuclear material (cell death) was remarkable morphologically, and the intracellular anticancer concentration of the conjugate was seven times higher than that of the anticancer drug alone (p less than 0.05). In conclusion, the conjugate has more cytotoxic effect than the anticancer drug alone against AFP-producing hepatoma cells in optimal conditions.

Deacetylravidomycin N-oxide, a new antibiotic. Taxonomy and fermentation of the producing organism and isolation, structure and biological properties of the antibiotic.

Streptomyces ravidus S50905 was found to produce a new antibiotic, deacetylravidomycin N-oxide, together with ravidomycin and deacetylravidomycin in a culture medium containing sodium anthraquinone-beta-sulfonate. The structure of this new compound was determined from NMR and mass spectrometric data, and further confirmed by chemical synthesis from deacetylravidomycin. Deacetylravidomycin N-oxide was antitumor active against P388 leukemia and Meth A fibrosarcoma in a wide range of doses, and considerably less toxic than deacetylravidomycin. Its antibacterial activity was less potent than deacetylravidomycin. Ravidomycin N-oxide was also synthesized from ravidomycin and its biological properties were tested.

[The significance of blood coagulation test in fulminant hepatic failure].

We treated 12 cases of fulminant hepatic failure with plasma exchange, cryofiltration and hemodialysis. Five cases were survived and 7 cases died. The liver volume measured by volus CT on admission was about 1106 ml in survived cases, and was about 457 ml in death cases. In cases which liver volume was under 600 ml, these prognoses were miserable. AT-III, PT and HPT activities were well correlated with liver volume (r = 0.85) and these activities were good indicator to treat the fulminant hepatic failure. The measurement of FBG and APTT were also useful for the estimation of the prognosis. But they were not becoming of indicator to treatment. We have no expectation to prognosis of the cases which PT, HPT and AT-III were under 15%, 10%, and 25%, respectively.