Translocation (3;21;8)(q21;q22;q22) in a patient with acute myeloid leukemia. A case report and review of prognostic indicators.

We report a patient with acute myeloid leukemia (AML) and t(3;21;8)(q21;q22;q22). This translocation has not been previously described in de novo or relapsed AML. The patient is a 25-year-old woman who presented with WBC 6.2 x 10(9)/L, Hgb 10.2 g/dL, Hct 28.4%, and platelets 67 x 10(9)/L. A bone marrow biopsy revealed a 70% hematopoietic cellularity with 65% blasts. Immunophenotyping showed aberrant expression of lymphoid-associated marker CD19. Cytogenetic analysis on a 72-hour culture of bone marrow cells supplemented with conditioned media was evaluated by G-banding at about the 400-band level. The patient's age, cytogenetics, WBC, and immunophenotype at diagnosis would seem to suggest a favorable prognosis, according to previous studies of prognostic indicators. She was treated with induction and consolidation chemotherapy, followed by myeloablative conditioning and autologous peripheral blood stem cell transplant (PBSCT). Despite multiple favorable prognostic factors, the patient relapsed 7 months after PBSCT. Translocation of chromosomes 8 and 21 is common in AML and is generally considered a good prognostic factor. We suspect that the effect of the 3q21 translocation in an otherwise favorable translocation of chromosomes 8 and 21 may be responsible for this patient's early relapse.

Interleukin 1beta and interleukin 6, but not tumor necrosis factor alpha, inhibit insulin-stimulated glycogen synthesis in rat hepatocytes.

Recent evidence indicates that inflammatory cytokines are involved in changes of blood glucose concentrations and hepatic glucose metabolism in infectious diseases, including sepsis. However, little is known regarding how cytokines interact with glucoregulatory hormones such as insulin. The objective of the present study is to investigate if and how cytokines influence insulin-stimulated glycogen metabolism in the liver. Interleukin 1beta (IL-1beta) and interleukin 6 (IL-6) markedly inhibited the increase of glycogen deposition stimulated by insulin in primary rat hepatocyte cultures; however, tumor necrosis factor alpha had no effect. Labeling experiments revealed that both cytokines counteracted insulin action by decreasing [14C]-glucose incorporation into glycogen and by increasing [14C]-glycogen degradation. Furthermore, it was discovered that IL-1beta and IL-6 inhibited glycogen synthase activity and, in contrast, accelerated glycogen phosphorylase activity. In experiments with kinase inhibitors, serine/threonine kinase inhibitor K252a blocked IL-1beta- and IL-6-induced inhibitions of glycogen deposition, as well as glycogen synthase activity, whereas another kinase inhibitor staurosporine blocked only IL-6-induced inhibition. Tyrosine kinase inhibitor herbimycin A blocked only IL-1beta-induced inhibition. These results indicate that IL-1beta and IL-6 regulate insulin-stimulated glycogen synthesis through different pathways involving protein phosphorylation in hepatocytes. They may mediate the change of hepatic glucose metabolism under pathological and even physiological conditions by modifying insulin action in vivo.

A simplified model of hypoxic injury in primary cultured rat hepatocytes.

The Anaeropack system for cell culture, which was originally designed for the growth of anaerobic bacteria, was used to produce a hypoxic atmosphere for cultured hepatocytes. We measured changes in the oxygen and carbon dioxide concentrations and the atmospheric temperature in an airtight jar. We also measured changes in the pH of the medium during hypoxia to assess the accuracy of this system. Moreover, we used three durations (2, 3, and 4 h) of hypoxia and 8 h of reoxygenation in cultured rat hepatocytes, and then measured the lactate dehydrogenase (LDH), ketone body concentration (acetoacetate + beta-hydroxybutyrate), and the ketone body ratio (KBR: acetoacetate/beta-hydroxybutyrate) in the medium in order to assess the suitability of this system as a model for reperfusion following liver ischemia. The oxygen concentration dropped to 1% or less within 1 h. The concentration of carbon dioxide rose to about 5% at 30 min after the induction of the hypoxic conditions, and was maintained at this level for 5 h. No effect of the reaction heat produced by the oxygen absorbent in the jar was recognized. The extent of cell injury produced by changing the hypoxic parameters was satisfactorily reflected by the KBR, the ketone body concentration, and the LDH activity released into the medium. Because this model can duplicate the conditions of the hepatocytes during revascularization following ischemic liver, and the Anaeropack system for cell culture is easy to manipulate, it seems suitable for the experimental study of hypoxic injury and revascularization in vitro.

Skin Necrosis Associated With Heparin-Induced Thrombocytopenia and Thrombosis.

Skin necrosis is a rare complication of heparin therapy. Strong evidence suggests an immune-mediated mechanism in which heparin-antibody complexes bind to platelets, resulting in platelet aggregation, thromboembolism, and ischemic necrosis. Heparin-induced thrombocytopenia (HIT) may also occur in response to immune-mediated platelet aggregation. The presence, of heparin-dependent antibodies can be confirmed by platelet aggregometry, (14)C-serotonin release assay (SRA), or enzyme-linked immunosorbent assay (ELISA). Clinical suspicion, early detection and immediate cessation of heparin therapy are important in preventing the potentially severe complications of heparin-induced platelet aggregation. Potential therapeutic approaches include plasmapheresis and alternative forms of anticoagulation such as warfarin, aspirin, dipyridamole, or other novel investigational agents.

[Selective thermocoagulation of unresectable malignant tumors using radiofrequency].

Based on our experimental findings on porcine liver, we have been conducting a clinical trial of selective hyperthermia by radiofrequency (RF) capacitive heating with laparotomy for patients with unresectable malignant tumors. In 10 patients with malignant tumors (8 carcinoma of the pancreas, 2 carcinoma of the gallbladder), laparotomy and RF heating were performed after informed consent. The local heat coagulation was produced by heating equipment using 13.56 MHz radiofrequency produced by Omron Corporation, Japan. Four 2-cm electrode needles were placed in the tumor in a square array at intervals of 2.0 cm. Hyperthermia was given for 30 min with a controlled temperature of 50 degrees C in the RF field (2 x 2 x 2 cm3). That of the surrounding area was maintained at less than 40 degrees C. The calculated volume treated by RF ranged between (2 x 2 x 2 cm3) x 1 and (2 x 2 x 2 cm3) x 6. We followed all patients by computed tomographic (CT) scan 2 weeks after coagulation. Tumor markers in the blood were assayed before and 14 days after heating. Follow-up CT scans demonstrated that after the tumor mass had been heterogeneously enhanced, it changed to a homogeneous low-density area in 6 of 10 patients. The levels of tumor markers decreased to lower than the pre-treatment values in 9 of 10 patients. In all patients, the changes in CT scans and/or decrease in the markers were confirmed. Complications such as bleeding or abscess formation were not observed. It was suggested that the selective hyperthermia was safely produced by this equipment. The encouraging results in these patients justify further clinical trials.

Regulation of energy metabolism by interleukin-1beta, but not by interleukin-6, is mediated by nitric oxide in primary cultured rat hepatocytes.

The effects of inflammatory cytokines (interleukin-1beta, interleukin-6, and tumor necrosis factor-alpha) on energy metabolism were studied in primary cultured rat hepatocytes. Adenine nucleotide (ATP, ADP, and AMP) content, lactate production, the ketone body ratio (acetoacetate/beta-hydroxybutyrate) reflecting the liver mitochondrial redox state (NAD+/NADH), and nitric oxide formation were measured. Insulin increased ATP content in hepatocytes and had a maximal effect after 8-12 h of culture. Both interleukin-1beta and interleukin-6, but not tumor necrosis factor-alpha, significantly inhibited the ATP increase time- and dose-dependently. Interleukin-1beta and interleukin-6 also stimulated lactate production. During the same period, interleukin-1beta but not interleukin-6 decreased the ketone body ratio. Furthermore, interleukin-1beta markedly stimulated nitric oxide formation in hepatocytes, and this increase was blocked by NG-monomethyl-L-arginine (a nitric oxide synthase inhibitor) and by interleukin-1 receptor antagonist. NG-monomethyl-L-arginine reversed inhibition of the ATP increase, decrease in the ketone body ratio, and increase in lactate production, which were induced by interleukin-1beta. Interleukin-1 receptor antagonist completely abolished all of the effects induced by interleukin-1beta. These results demonstrated that interleukin-1beta and interleukin-6 affect the insulin-induced energy metabolism in rat hepatocytes by different mechanisms. Specifically, interleukin-1beta inhibits ATP synthesis by causing the mitochondrial dysfunction, a process which may be mediated by nitric oxide.

Adenylate energy charge of rat and human cultured hepatocytes.

A simple and rapid method for the assay of adenine nucleotides (ATP, ADP, and AMP) was established to evaluate the adenylate energy charge (ATP+ADP/2)/(ATP+ADP+AMP) of cultured hepatocytes. The effects of inhibitors of glycolysis, fatty acid oxidation, or oxidative phosphorylation on the energy charge were examined. The energy charges of cultured hepatocytes in rats and human were almost identical and were maintained at a high level between 6 and 24 h after changing the media (rat: 0.908 +/- 0.008 n = 9, human: 0.918 +/- 0.014 n = 6, mean +/- SD). Inhibition of glycolysis with sodium fluoride or oxidative phosphorylation with antimycin A irreversibly reduced both the adenine nucleotide contents and the energy charge. However, the inhibition of fatty acid oxidation with 2-tetradecylglycidic acid did not affect the nucleotide contents, and the energy charge only decreased transiently to recover within 8 h. When the inhibitor of oxidative phosphorylation was removed, the recovery in the energy charge preceded the recovery in the adenine nucleotide contents. These findings suggest that the adenylate energy charge is a more sensitive measure of the changes in energy metabolism than the adenine nucleotide contents. Furthermore, energy charge regulates adenine nucleotide contents in cultured hepatocytes. It is important to confirm that the high energy charge of the cultured hepatocytes is maintained when these cells are used for metabolic studies.

[Influence of epinephrine infusion on the metabolism of exogenous lipid emulsions after 70% hepatectomy in the rat].

We investigated the effects of epinephrine on the metabolism of a lipid emulsion administrated to rats after 70% hepatectomy. Sprague-Dawley rats were underwent 70% hepatectomy and then were given lipid total parenteral nutrition (lipid-TPN) for 96 h. To increase catabolism after hepatectomy, epinephrine (Bosmine) infused continuously with the lipid-TPN. With increasing doses of the epinephrine, weight loss and the increased urinary excretion of nitrogen and 3 methyl histidine were observed. This indicated that administration of epinephrine after hepatectomy produced a model of increased surgical stress. As the epinephrine dose increased the linoleic acid content of the hepatic phospholipids increased and the arachidonic acid decreased at 96 h after hepatectomy. Epinephrine apparently inhibited the activity of the delta 5.6 desaturase involved in the biosynthesis of polyunsaturated fatty acids, leading to changes of the cell membrane fatty acid composition that could possibly influence membrane function. Thus, the fatty acid composition of lipid emulsions for TPN should be tailored according to the severity of surgical stress, because the pattern of secretion of stress hormone is dependent on the degree of such stress.

Stimulation of glycogen degradation by prostaglandin E2 in primary cultured rat hepatocytes.

Hepatocytes isolated from rats by the collagenase perfusion method were cultured as monolayers at concentrations of 0.4-1.1 x 10(6) attached cells/dish (9 cm2) for 1-3 days and the effect of prostaglandins on their glycogenolysis was studied. By use of [14C]glycogen-labeled cells, prostaglandin E2 (PGE2) was found to have a stimulatory effect on glycogen degradation at high cell density (more than 0.8 x 10(6) cells/dish) in 1-day cultures. PGE2 was maximally effective at 10(-7) M, increasing [14C]release from cellular [14C]glycogen to 2-3 times the basal level after 1 h incubation, and to plateau level within 2 h. PGE1, 16,16-dimethyl PGE2 and PGF2 alpha had similar effects, but PGD2 and dinor-PGE1 (a metabolite of PGE1 and PGE2 in hepatocytes) had no effect. This prostaglandin-induced glycogen degradation was observed in 1-day cultures, with a maximum between 20-30 h, but not in 2-day and later cultures. Treatment of hepatocytes with pertussis toxin potentiated PGE2-stimulated glycogen degradation, indicating that the effect involves a different pathway from that for inhibition of glucagon- and epinephrine-stimulated glycogenolysis by E series prostaglandins reported previously.

Stimulation of glucose incorporation into glycogen by E-series prostaglandins in cultured rat hepatocytes.

In primary cultures of rat hepatocytes, 16,16-dimethylprostaglandin E2 (16,16-dimethyl PGE2), a biologically active analogue of prostaglandin E2 (PGE2), stimulated the basal rate of [14C]glucose incorporation into glycogen. 16,16-Dimethyl PGE2 caused concentration-dependent stimulation (ED50: 10(-8) M) with a maximum 2-3 h after its addition. Prostaglandin E1 (PGE1), PGE2 and prostaglandin F2 alpha (PGF2 alpha) stimulated also the incorporation, but less effectively than 16,16-dimethyl PGE2. However, prostaglandin D2 (PGD2) did not show such effect. Cellular glycogen analysis revealed that PGE2 and 16,16-dimethyl PGE2 increased a net glycogen accumulation time-dependently. Pretreatment of the cultured hepatocytes with pertussis toxin blocked the effects of PGE2 and 16,16-dimethyl PGE2 completely and concentration-dependently. These findings indicate that E-series prostaglandins have significant effects on hepatic glycogenesis via pertussis-toxin-sensitive G protein, in addition to their inhibitory effects on hormone-stimulated glycogenolysis reported previously (Okumura, T., Sago, T. and Saito, K. (1988) Prostaglandins 36, 463-475).

The usefulness of intraoperative drip infusion cholangiography during laparoscopic cholecystectomy.

Intraoperative cholangiography during laparoscopic cholecystectomy has been considered to be a necessary examination because incidental injury to the common bile duct must be avoided. We performed 93 intraoperative drip infusion cholangiographies among 103 laparoscopic cholecystectomized patients as simple examinations by using iotroxic acid. The best drip infusion time was determined to be 20 min and good pictures were obtained from 10 to 60 min after the end of the drip. Nine patients with liver dysfunction and a poor radiograph had poor cholangiograms. Clear cholangiograms were obtained in 79 patients: four had a long remnant cystic duct and, in one case, a common bile duct stenosis was found by endoclip. The findings in these five cases helped us to correct failures during operation.

Branched-chain amino acid-supplemented nutritional support after gastrectomy for gastric cancer with special reference to plasma amino acid profiles.

Preoperative plasma aminograms constructed for 63 patients expected to undergo gastrectomy for gastric cancer at different stages showed markedly lower concentrations of many plasma amino acids in the Stage IV and recurrent cases. The amino acid levels were inversely proportional to tumor size. On the other hand, preoperative arteriovenous differences in free amino acid levels were positive in Stage I cancer but negative in Stage IV cancer, indicating that intake of amino acids by the skeletal muscles exceeded the outflow in Stage I, whereas there was a net loss of amino acids from the skeletal muscles in advanced cancer. The amount of amino acids actually lost from the skeletal muscles after muscular loading in Stage I cancer also surpassed that in Stage IV cancer. Administration of TPN solution supplemented with 31% branched-chain amino acids (BCAA) might favorably influence muscle protein metabolism in gastric cancer patients by inhibiting protein degradation and promoting synthesis, as treatment was more effective than 21% BCAA-enriched TPN.

Preoperative nutritional assessment to predict postoperative complication in gastric cancer patients.

The correlation between preoperative nutritional parameters and postoperative complications in 440 patients with gastric cancer were analyzed. All the nutritional parameters reflected a significant deterioration as the stages of cancer progressed, and the frequency of postoperative complications was highest in patients with stage IV gastric cancer. The incidence of anastomotic leaks was increased in patients undergoing total gastrectomy with no relation to the clinical stage or nutritional status. However, there was a close relationship between nutritional status and immunocompetence, lung complications, and infection. The nutritional indices which reliably predicted preoperatively the nutritional status of cancer patients were the serum protein concentrations including the serum albumin (Alb) and prealbumin (PA). The indices predicting postoperative complications were the Alb, PA, and total lymphocyte count. These results suggest that preoperative nutritional assessment can be beneficial for the prediction of postoperative complications.