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The authors describe 2 patients who presented with transient directional disorientation (TDD) as a manifestation of cerebral ischemia. The patients suddenly lost sense of direction in a familiar environment despite preserved ability to recognize landmarks, and recovered within a short time. Brain MRI revealed an ischemic lesion in the right medial occipital lobe and the corpus callosum in case 1 and in the right parieto-occipital sulcus (POS) in case 2. After ictus, fMRI study of a navigation task was performed, which demonstrated the activation of the POS unilaterally in case 1 and bilaterally in case 2. We propose that TDD of our patients is related to temporary dysfunction of bilateral POS., ((c) 2008 S. Karger AG, Basel.)

Histone deacetylase (HDAC) inhibitors are promising candidates for molecular-targeted therapy for leukemia. In this study, we investigated the mechanisms of cytotoxic effects of depsipeptide (FK228), one of the most effective HDAC inhibitors against leukemia, using human myeloid leukemic cell lines HL-60 and K562. We found that FK228 activated caspase-9 and a subsequent caspase cascade by perturbing the mitochondrial membrane to release cytochrome c, which was almost completely blocked by overexpression of Bcl-2. The mitochondrial damage was caused by the translocation of Bax but not other pro-apoptotic Bcl-2 family proteins to the mitochondria. FK228 did not affect the interaction between Bax and Bax adaptor proteins such as 14-3-3theta and Ku70. FK228-induced apoptosis and mitochondrial translocation of Bax were markedly enhanced by the proteasome inhibitor bortezomib. The synergistic action of FK228 and bortezomib was at least partly mediated through conformational changes in Bax, which facilitate its translocation to the mitochondria. These results suggest that the combination of HDAC inhibitors and proteasome inhibitors is useful in the treatment of leukemia especially in the context of molecular-targeted therapy. The status of Bcl-2 and Bax may influence the sensitivity of tumors to this combination and thus can be a target of further therapeutic intervention., (Copyright 2006 S. Karger AG, Basel.)

Background/aim: Gene transfer into the kidney has great potential as a novel therapeutic approach. However, an efficient method of gene transfer into the kidney has not been established. We explored the transduction efficiency of renal cells in vitro and in vivo using adeno-associated virus (AAV) serotype 1-5 vectors encoding the beta-galactosidase gene., Methods: In the in vitro study, rat kidney epithelial cell line NRK52E cells were transfected with AAV serotype derived vectors. In the in vivo study, AAV serotype derived vectors were selectively injected into the kidney using a catheter-based gene delivery system in rats and mice mimicking the clinical procedure. The efficiency of gene expression was histologically evaluated on the basis of the beta-galactosidase expression., Results: AAV serotype 1, 2, and 5 vectors transduced in rat kidney epithelial cell line NRK52E cells in vitro, whereas AAV serotype 3 or 4 vectors showed no transduction. In addition, the kidney-specific injection of AAV serotype 2 vectors successfully transduced in tubular epithelial cells, but not in glomerular, blood vessel, or interstitial cells in vivo, whereas the rest of the serotypes showed no transduction., Conclusion: Since kidney-specific gene delivery via the renal artery by catheterization is highly feasible in humans, these findings provide useful information for promising strategies in renal gene therapy., (Copyright 2004 S. Karger AG, Basel)

It has been shown that there is an inverse relationship between the level of thymidylate synthase (TS) and therapeutic outcomes in patients with malignancies including cervical cancer. To clarify the mechanism of the poor prognosis of cervical cancer with high TS expression, we introduced TS cDNA to the human uterine cervical cancer cell line SKG-II and evaluated the effect of TS expression on its radiosensitivity. After selection, stable transformants of SKG-II cells expressing high level of TS (SKG-II/TS) and control cells (SKG-II/luciferase) were obtained. The level of TS measured by the FdUMP-TS binding assay was significantly higher (p < 0.05) in SKG-II/TS than in control (2.0 +/- 0.1 and 1.3 +/- 0.1 pmol/mg, respectively). No difference was observed in in vitro cell growth or in vivo tumor growth. On evaluation of in vitro radiosensitivity, the 50% growth inhibitory dose (ID(50)) was 3.1 +/- 0.1 Gy in SKG-II/TS and was significantly higher (p < 0.01) than that in control (2.2 +/- 0.1 Gy). From these results, it is suggested that one of the reasons of poor outcome of cervical cancer to radiation is high TS expression., (Copyright 2002 S. Karger AG, Basel)

Adeno-associated virus (AAV) vectors have a number of attractive features, including lack of cytotoxicity, ability to transduce nondividing cells, and long-term transgene expression. We investigated whether rat renal cells could be efficiently transduced with AAV vectors. Rat glomerular mesangial cells were transduced with AAV-lacZ vector containing beta-galactosidase gene in vitro, and the expression of beta-galactosidase was evaluated by X-gal staining and ELISA. For ex vivo experiments, sections of rat kidneys were incubated with AAV-lacZ, and then evaluated by X-gal histochemical staining. The level of beta-galactosidase expression in cultured rat mesangial cells increased in a dose-dependent manner (ranging from 1 x 10(5) to 5 x 10(6) particles/cell). When transduced with 5 x 10(6) vector particles/cell of AAV-lacZ, about 50% of mesangial cells were stained positively with X-gal, and the level of beta-galactosidase expression reached 9.9 +/- 1.5 ng/mg protein. Expression was detectable during the culture period for at least 7 days. X-gal histochemical examination of the ex vivo transduced renal tissue revealed tubular cell and interstitial tissue staining. However, gene transfer was not clearly observed in glomeruli. These findings suggest that AAV vectors have the potential for gene therapy of renal diseases., (Copyright 2000 S. Karger AG, Basel)

Glycoprotein B (gB) of human cytomegalovirus (CMV) is the major target protein of neutralizing antibodies, and four variant types are known. A previously reported polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method was modified by eliminating the cell culture step and amplifying a CMV fragment (corresponding to a.a. 441-450 of gB) by nested PCR. DNA was extracted from the sera of 27 pediatric bone marrow transplant recipients and subjected to nested PCR. Of the samples, 20 yielded PCR products, and the gB type was determined by RFLP. Of the 20 patients, 4 (20%) had gB type 1 (Towne type), 15 patients (75%) had gB type 2 (AD 169 type), and 1 patient (5%) had gB type 3. Previous studies showed that gB type was most common among Caucasians [Chou and Dennison: J Infect Dis 1991;163:1229-1234; Fries et al: J Infect Dis 1994;169:769-774]. Thus, gB genotypes seem to be distributed differently in Caucasians and Japanese. Further, nucleotide sequence analysis of the amplified region revealed that all the type 2 viruses had the same amino acid sequences. The type 3 sample had a novel amino acid substitution at position 498. Of the type 1 samples, 3 had amino acid substitutions in various positions: 1 sample at position 493, 1 sample at position 447 and 1 sample at position 452, 493 and 498.

The protective effect of a novel prostacyclin (PGI2) analog, OP-2507, on mesenteric circulation was investigated in a canine warm ischemia model. In 20 mongrel dogs, the entire portion of the intestine supplied by the superior mesenteric artery (SMA) and drained by the superior mesenteric vein (SMV) was completely isolated, maintaining the blood and lymph vessels intact. Sixty or 120 min of complete warm ischemia (WI) of the intestine was induced by clamping SMA and SMV, followed by reperfusion for 120 min. Animals were divided into five groups (each n = 4): group 1, sham operation; group 2, 60 min WI; group 3, 120 min WI; group 4, 60 min WI with PGI2 analog administration; group 5, 120 min WI with PGI2 analog administration. The analog was administered at a rate of 6 micrograms.kg-1.h-1 immediately after laparotomy until the end of the observation period. Mean arterial pressure, SMA blood flow (SMABF), SMV pressure were monitored and total mesenteric vascular resistance (TMVR) was calculated. To evaluate the endothelial activation, endothelin, which is secreted from the endothelium under hypoxic stress, was assayed from blood samples of SMV. None of the animals showed significant changes in mean arterial pressure. In groups 2 and 3, SMABF decreased significantly to less than 60% of preoperative value (15 ml.kg-1.min-1) and TMVR significantly increased from 8.1 and 7.3 mm Hg.ml-1.kg.min before WI to 14.0 and 16.4 mm Hg.ml-1.kg.min after 120 min reperfusion, respectively, resulting in delayed hypoperfusion. In contrast, in groups 4 and 5, SMABF increased to over 100% of preoperative level, while TMVR declined from 7.8 and 8.4 mm Hg.ml-1.kg.min before WI to 6.2 and 6.3 mm Hg.ml-1.kg.min after 120 min reperfusion. After 60 min reperfusion, SMABF and TMVR showed a significant difference between the treated and nontreated groups. Only in group 3, high endothelin concentrations (over 20 pg/ml) were observed even after 120 min reperfusion. It was concluded that the PGI2 analog was able to suppress the endothelial activation and the disturbance of mesenteric circulation caused by WI and reperfusion.

Growth hormone (GH), which is well known as an anabolic agent in systemic protein metabolism but has catabolic effects on the carbohydrate metabolism in the liver, was administered to normal rabbit to investigate its effects on the hepatic energy metabolism. The changes in arterial ketone body ratio (AKBR: acetoacetate/3-hydroxybutyrate), which reflects the hepatic mitochondrial redox state ([NAD+]/[NADH]), after GH injection was studied as an indicator of the hepatic energy metabolism. GH was administered to normal rabbit at the doses of 50 micrograms/kg (GH-50 group), 100 micrograms/kg (GH-100 group) and 200 micrograms/kg (GH-200 group) by intravenous bolus injection. In the GH-50, GH-100 and GH-200 groups, AKBR decreased significantly from 1.40 +/- 0.09 to 0.94 +/- 0.05, from 1.19 +/- 0.11 to 0.83 +/- 0.14, and from 1.19 +/- 0.08 to 0.71 +/- 0.15 at 90 min, respectively. The energy charge of the liver decreased significantly 90 min after 200 micrograms/kg GH injection from 0.872 +/- 0.003 to 0.836 +/- 0.012 (p < 0.05). These results suggest that GH is associated with the deterioration of the hepatic energy metabolism, and that the administration of GH should be carefully weighed up in cases of damaged liver.

To clarify the mechanism for Cl- transport in swine tracheal submucosal gland cells, we measured chloride (Cl-) flux by means of a Cl(-)-sensitive fluorophore, 6-methoxy-N-(3-sulfopropyl)quinolinium (SPQ). An abrupt change of Cl(-)-free bath to a Cl(-)-containing (114 mM) solution induced the Cl- influx into the cells at a rate of 0.52 mM/s (p < 0.05). Removal of sodium (Na+) and the addition of 0.5 mM furosemide significantly reduced the rate of gradient-induced Cl- influx to 0.21 (p < 0.05) and 0.19 mM/s (p < 0.05), respectively. In gland tissue treated with 0.1 mM ouabain, the removal of external potassium (K+) in the presence of 5 mM barium significantly reduced the rate of Cl- influx to 0.21 mM/s (p < 0.05). An abrupt change from a Cl(-)-containing bath to a Cl(-)-free solution induced the Cl- efflux from the cells at a rate of 0.10 mM/s (p < 0.05). Acetylcholine increased the rate of gradient-induced Cl- efflux in a dose-dependent manner to 0.78 mM/s at 10(-6) M. This effect of acetylcholine was significantly diminished by diphenylamine-2-carboxylic acid (DPC, 10(-9) M), a Cl- channel blocker (p < 0.05). Isoproterenol (10(-5) M) had no effect on the Cl- efflux. These findings indicate that a Na-K-Cl co-transporter plays a major role in Cl- entry, and that Cl- efflux was activated by cholinergic receptor stimulation, but not by beta-adrenergic stimulation, via a DPC-inhibitable Cl- channel.

The effect of the gabexate mesilate (Gab) on thrombin and plasmin generation following liver resection in cirrhotic patients was studied. Six cirrhotic patients received an infusion of Gab after liver resection (Gab group), and another 6 patients did not receive such treatment (Con group). The parameters measured were thrombin-antithrombin complex (TAT), plasmin-antiplasmin complex (PAP) and D-dimer. The real increases of D-dimer and PAP were significantly higher in Con group after surgery while no significant difference was observed in the increase of TAT. These results show that Gab suppresses plasmin generation and following D-dimer production more effectively than thrombin generation after hepatic resection.

We measured lymphocyte energy charge (LEC) in septic patients after hepatectomy to clarify the energy metabolism of lymphocytes in relation to arterial ketone body ratio (AKBR) reflecting the hepatic mitochondrial redox potential. Sixteen patients with AKBR above 0.7 (state A) tolerated their operations well without postoperative infectious episodes and their LEC (0.895 +/- 0.005, mean +/- SEM) was significantly (p < 0.001) higher than that (0.841 +/- 0.010) of 9 patients with AKBR from 0.7 to 0.4 (state B). Four of the 9 state B patients had multiple organ failure (MOF) with sepsis as a trigger. AKBR in 7 of 9 state B patients decreased and remained below 0.4 (state C). These state C patients showed significantly reduced LEC (0.781 +/- 0.024; p < 0.001, p < 0.05, compared with that in states A and B, respectively) and finally died of MOF with septic state. These results suggest that the severe and prolonged impairment of energy metabolism in the liver may be accompanied with the metabolic derangement of lymphocytes.

In liver transplantation, graft viability is ideally to be determined before implantation. Integrity of mitochondria may be a prerequisite to a viable graft. A new method is presented, which allows for the determination of the membrane potential of mitochondria (MPM; mV) in state 4 respiration within 50 min in 40-mg specimens, employing rhodamine 123 as a probe. Normal control showed a MPM of 239.2 mV. Storage in saline at 37 degrees C yielded an impaired MPM of 153.5 mV within 3 h. The cold storage at 1 degree C could preserve MPM at quasi-normal after 3 h but reduced it significantly after 24 h to 222.2 mV in saline (p < 0.005 vs. control) and 231.0 mV in UW solution (p < 0.05 vs. control): the difference between the 24-hour values was significant (p < 0.05).

The aim of this study is to investigate the relationship in energy metabolism between the platelet and the liver. The adenylate energy charge of human platelets and arterial ketone body ratio (AKBR:acetoacetate/3-hydroxybutyrate) were measured in 11 patients after hepatectomy. Hepatic energy crisis was defined as the decrease of AKBR below 0.7. The platelet energy charge was measured on the first (Day 1), the second (Day 2), third (Day 3) and sixth (Day 6) day during AKBR decreased and remained under 0.7. The values before AKBR decreased were employed as the control. The values of the platelet energy charge were 0.917 +/- 0.008 in the control (n = 11) and 0.896 +/- 0.009 in all samples after AKBR decreased under 0.7 (n = 30). There was no significant difference between them. The values of energy charge were divided into four groups according to the periods of time after AKBR decreased and remained under 0.7 and compared. The energy charge was 0.923 +/- 0.006 (Day 1, n = 11), 0.907 +/- 0.008 (Day 2, n = 10), 0.890 +/- 0.005 (Day 3, n = 5), and 0.815 +/- 0.012 (Day 6, n = 4). The energy charge of Day 3 was significantly lower than that of Day 1 (p < 0.01). The energy charge of Day 6 was significantly lower than that of Day 1, 2, 3 (p < 0.001, p < 0.001, and p < 0.01, respectively). The 4 patients whose AKBR remained under 0.7 for more than 6 days had hemostatic disorder. The deterioration of the platelet energy charge metabolism lags behind that of the liver.(ABSTRACT TRUNCATED AT 250 WORDS)

The influence of deteriorated liver function on the healing process of arterial anastomosis was investigated in rats who underwent arterial reconstruction by the microscopic surgical techniques of conventional sutured vascular anastomosis (CSVA) and laser-assisted vascular anastomosis (LAVA). After 70% hepatectomy, the arterial ketone body ratio, reflecting the hepatic mitochondrial redox state, and the adenylate energy charges of the platelets, polymorphonuclear leukocytes and mononuclear cells, which play an important role in wound healing, were significantly lower in hepatectomized rats than in control. Tensile strength and bursting pressure with CSVA at the abdominal aortic anastomosis were also significantly lower in hepatectomized rats than in control. While CSVA was superior to LAVA in tensile strength, bursting pressure and incidence of aneurysm formation in nonhepatectomized rats, LAVA tended to show a lower degree of disadvantage in hepatectomized rats than CSVA, especially in the incidence of calcification at the anastomotic site. It was concluded that LAVA would be a useful option in vascular anastomosis, even during extended hepatectomy.

The activities of ATP-synthetase and ATPase in rabbit liver were evaluated by kinetic analysis of rapid changes in ATP during alteration of oxygen supply, and were compared between normal and hepatectomized rabbits. Velocity constants of ATP-synthetase and ATPase were computed on the assumption that ATP-synthetase and ATPase follow the pseudo-first order reaction of ADP and ATP, respectively. The velocity constant of ATPase increased from the control value of 0.21 to 0.48 (min-1) in the remnant rabbit liver at 24 h after 70% hepatectomy. The velocity constant of ATP-synthetase increased from the control value of 4.05 to 5.24 (min-1) after the hepatectomy. These results indicate that both ATP-synthetase and ATPase are accelerated due to liver regeneration.

Porcine livers were preserved for 72 h using continuous hypothermic perfusion with UW solution and fluorocarbon emulsion (FC-43). They were then recirculated with human blood for 2 h at 37 degrees C for the evaluation of their viability. During hypothermic perfusion and recirculation, the main metabolites of the perfusate and adenine nucleotide concentrations were measured in order to examine the energy metabolism of the livers. This study revealed that hypothermically perfused livers, which mainly utilize endogenous fatty acids as an energy substrate during this time were adequately preserved for 72 h. In terms of energy metabolism, it is suggested that when using the method of continuous hypothermic perfusion with UW solution and FC-43, the administration of adequate amounts of certain fatty acids might prolong the preservation period.

A few cases of nephrotic syndrome with the glomerular deposition of an amyloid-like material which did not stain with Congo red have been documented. But extrarenal deposits have not been previously reported in this disease. We describe here a case of nephrotic syndrome associated with the deposition of an amyloid-like material in the liver as well as in the renal glomeruli. The deposits were made up of fibrillar structures which resembled those of amyloid when viewed through the electron microscope but they did not stain with Congo red. This is the first report of amyloidosis-like glomerulopathy with extrarenal deposits.

Rat orthotopic liver transplantation was performed using a newly synthesized bioabsorbable material (LA-GA copolymer) cuffs and the ordinary polyethylene cuffs. The LA-GA copolymer cuff which anastomosed the portal vein was patent and developed no collateral veins even after 6 months, keeping the transplanted liver normal. By contrast, the polyethylene cuff-anastomosed portal vein was completely occluded and the collateral veins were highly developed, with the transplanted liver showing the fatty degeneration of hepatocytes and numerous regenerative nodules. It is concluded that the LA-GA copolymer cuff is a suitable material for the short- and long-term study of rat orthotopic liver transplantation.

The effect of a new oil-soluble anticancer agent 'KM2210' (estradiol-chlorambucil) dissolved in lipiodol (LPD) was investigated as intraarterial chemotherapy, using 40 Wistar rats bearing Walker 256 carcinosarcoma in the extremities. KM2210-LPD significantly inhibited tumor growth and prolonged the survival time, as compared to LPD alone or saline alone, p less than 0.01 and p less than 0.01, respectively. Pathological study revealed that KM2210-LPD made the tumor necrotic. It was revealed that KM2210-LPD injected into the femoral artery was retained in the hind limb tumor. These findings suggest that KM2210 may possibly become a new anticancer agent with potential clinical use in LPD chemoembolization.

To clarify the mechanism of metabolic derangement by massive blood transfusion to the damaged liver, the changes in the hepatic mitochondrial redox state, as reflected in arterial ketone body ratio (acetoacetate/3-hydroxybutyrate), were studied in jaundiced rabbits by infusion of massive citrate-phosphate-dextrose (CPD) solution. The jaundiced rabbits received common bile duct ligation (BDL group), and the sham group had a simple exploration of the bile duct. CPD solution was infused for 3 h at a rate of 9 ml/kg/h in each group. As metabolic parameters, blood gas, pyruvate, lactate, citrate and ketone body ratio were analyzed in the arterial blood. During the course, arterial ketone body ratio decreased significantly (p less than 0.01) in the BDL group with severe metabolic acidosis, while it was maintained at high value in the sham group with metabolic alkalosis. Organic acids were more highly accumulated in the BDL group than in the sham group. These results suggested a hepatodepressant effect of massive blood transfusion, especially in the damaged liver.

A total of 205 studies of hemodynamics and blood gas analyses were performed in relation to hepatic mitochondrial redox state in 23 partially hepatectomized patients. When the blood ketone body ratio (the ratio of acetoacetate to beta-hydroxybutyrate in arterial blood) decreased, that is when the hepatic mitochondrial redox state was reduced, a hyperdynamic state with increased cardiac index, decreased systemic and pulmonary vascular resistance together with decreased oxygen consumption was observed in spite of the increased oxygen availability. Inhibited oxidative metabolism in the Krebs cycle may have resulted in the decreased oxygen consumption and decreased vascular tone. This study supports the concept that the hepatic metabolic derangements due to reduced hepatic mitochondrial redox state may be a factor in the decreased oxygen consumption and hyperdynamic state.

Ciclosporin (CsA) was administered subcutaneously at a dose of 3 mg/kg body weight/day from the day of operation to 14 days of liver allotransplantation in ACI rat (RT1a) to LEW rat (RT1(l) strain combination. All LEW recipients of ACI liver transplants without immunosuppressive treatment had severe rejection and expired within 12 days. In contrast, 7 out of 9 recipients in the same strain combination with temporary CsA treatment survived indefinitely. Histologically, widespread cellular infiltration and massive hepatocyte necrosis were evident upon autopsy of the recipients without CsA treatment. In contrast, in the surviving rats of the CsA-treated group, mononuclear cell infiltration was restricted to the periportal field and hepatocytes appeared to be normal at 14 days posttransplant. CsA concentrations in whole blood were determined by high-performance liquid chromatography. The trough levels were 788 +/- 48, 621 +/- 76 and 546 +/- 52 ng/ml, at 5, 10 and 14 days posttransplant, respectively. We concluded that this relatively low-dose subcutaneous administration of CsA offered adequate immunosuppression in rat liver allotransplantation in this strain combination.

Glucose metabolism and plasma amino acid pattern were investigated in relation to arterial blood ketone body ratio (acetoacetate/3-hydroxybutyrate), which reflects liver mitochondrial redox state, in rabbits subjected to graded hepatectomy (25, 70 and 93%) and 70% hepatectomy with common bile duct obstruction. After partial hepatectomy, blood ketone body ratio decreased according to the extent of resected mass. Common bile duct ligation induced a further decrease in blood ketone body ratio in 70% hepatectomized rabbits. During a 6-hour hypertonic glucose infusion (0.7 g glucose/kg/h) which started 24 h after the operation, the blood glucose level remained between 230 and 280 mg/dl in 25 and 70% hepatectomized groups, while it increased linearly in 93% and jaundiced 70% hepatectomized groups. The plasma concentrations of alanine and proline as well as other amino acids increased as the blood ketone body ratio decreased. It is suggested that Krebs cycle and gluconeogenesis activities are inhibited as blood ketone body ratio decreases markedly resulting in an impaired glucose tolerance and amino acid imbalance.

Acute changes associated with anhepatic and revascularized state were analyzed in relation to mitochondrial energy metabolism in rabbits. Fifteen minutes after the induction of anhepatic state, the ketone body ratio in the arterial blood decreased markedly from 0.833 to 0.229. The ketone body concentration also decreased to 24% of the normal value. The blood ketone body ratio and concentration remained low during the next 45 min. Pyruvate and lactate levels and the pyruvate/lactate ratio in the arterial blood changed very little during the 60-min anhepatic state. In a revascularization model following a 15-min anhepatic state, the ketone body concentration rose rapidly and then declined gradually, finally attaining a constant level at 30 min. The decreased blood ketone body ratio recovered to 0.55 at 30 min, and achieved steady state thereafter. Mitochondrial phosphorylative activity and hepatic energy charge showed almost normal levels 60 min after revascularization following the 15-min anhepatic state. These results indicate that the rapid metabolic alterations associated with the anhepatic and revascularized state can be accurately determined by measuring the arterial blood ketone body ratio, and they suggest that the ketone body ratio may serve as a convenient parameter for the evaluation of liver viability in pathological states such as operative ischemia or liver transplantation.

Interference between two different in vivo antigen-antibody reactions which occurred simultaneously was studied in guinea pigs. Animals were presensitized with an aqueous fraction (CP) extracted from cells of Corynebacterium equi, strain KO-85. Skin reactions of the Arthus type elicited by the CP fraction inhibited passive cutaneous anaphylaxis (PCA) induced by bovine serum albumin (BSA) and rabbit anti-BSA serum. PCA reactions of the CP-sensitized animals were also inhibited upon simultaneous challenges of BSA and the CP as evidence by reduction of PCA titer of the anti-BSA serum.

Changes in plasma proteins and hematocrit (Ht) were studied in jaundiced rabbits subjected to hemorrhagic hypotension induced by the modified method of Wiggers and compared with changes in the hepatic energy status. At 48 h after common bile duct ligation, jaundiced rabbits showed hypoalbuminemia and a lower hepatic energy charge (EC), (ATP+1/2ADP)/(ATP+ADP+AMP). 30 min after induction of hypovolemic hypotension, in order to maintain a mean arterial pressure of 40 mm Hg, 86% of the blood collected in the reservoir (about 19% of the normal whole blood volume) had to be successively reinfused over a 2-hour period. Despite increases in Ht and plasma total protein (p-TP) occurring in parallel with the reinfusion, plasma albumin (p-Alb) and the ratio of p-Alb to plasma globulin (A/G) did not increase. On the other hand, in sham-operated rabbits small amounts of blood had to be frequently removed during the first hour (about 34% of the normal whole blood volume), and only 34% of it had to be reinfused over a 2-hour period. Despite removal of more blood and a concomitant decrease in Ht, p-Alb and A/G ratio were higher than in jaundiced rabbits. After marked decreases in EC and total adenine nucleotides (TAN) due to the initial blood loss, EC recovered somewhat and remained higher in sham-operated rabbits than in jaundiced rabbits, whereas TAN did not recover in either group. This seems to indicate that hemodilution functions effectively to maintain microcirculation following hemorrhagic hypotension in sham-operated rabbits, while in jaundiced rabbits a rapid breakdown of protective homeostatic mechanisms results in the earlier deterioration of the hepatic energy status.

The effects and safety limits of acute hemodilution on hepatic energy status were investigated in relation to arterial blood ketone body ratio and hepatic energy charge in a hemodilution rat model. As long as the hematocrit value was maintained above 20%, ketone body ratio and energy charge level at 6 h after hemodilution remained at the same levels as those of the sham-diluted groups. However, when hematocrit value was less than 15%, the ketone body ratio markedly decreased from the control value of 0.686 +/- 0.044 to 0.278 +/- 0.048 (p less than 0.001), and energy charge decreased from the control value of 0.856 +/- 0.012 to 0.0806 +/- 0.011 (p less than 0.01). From these results, it was suggested that hemodilution exerts no influence on the energy status of the liver as long as hematocrit is maintained above 20%.

Changes in the composition of serum bile acids were investigated after orthotopic liver transplantation in piglets. Serum levels of all bile acid components rapidly increased during the anhepatic phase and then quickly decreased, returning to the preoperative state within 6 h of revascularization of the allograft. In the animals in which extrahepatic biliary stenosis was a complication, serum total bile acids (TBA) increased remarkably. A marked increase of hyocholic acid and marked decrease of hyodeoxycholic acid were noted; as percentages of TBA, the average levels per day were 91.3 +/- 1.3 and 5.0 +/- 1.3%, respectively. No change in chenodeoxycholic acid (CDCA) was observed. In the animals suffering acute rejection without extrahepatic biliary stenosis, serum TBA increased slightly and the percentage of CDCA rose, the average level being 29.6 +/- 1.5%. These results suggest that the measurement of the composition of serum bile acids may serve as a useful means of assessing allograft function after liver transplantation.

In previous studies, we analyzed the hemopoietic precursor cells of tumor-bearing nude mice with marked neutrophilia induced by a transplanted human lung cancer. In this study, we examined the morphology of granulocyte-macrophage colonies using a technique for cytochemical examination of all colonies grown in agar gel culture. By stimulation with spleen cell-conditioned medium, the bone marrow cells and spleen cells of tumor-bearing nude mice formed a higher proportion of granulocyte-containing colonies than those of control mice. However, a larger number of granulocyte-macrophage progenitors were not definitely committed to differentiate into granulocytes and macrophages, as no significant differences in cellular composition of colonies between tumor-bearing mice and control mice were observed by stimulation with L cell-conditioned medium.

Two patients with IgA nephropathy and a patient with Henoch-Schönlein purpura nephritis each associated with anterior uveitis are described. As anterior uveitis accompanying IgA nephropathy improved, renal manifestations were relieved. The patient with Henoch-Schönlein purpura nephritis suffered from not only anterior uveitis but also keratitis. It is suggested that immune mechanisms which induce IgA nephropathy may play a role in the development of anterior uveitis and keratitis.

The early metabolic disturbances in the liver following ligation of the left hepatic or the common duct were studied in the rabbit. In the rabbits subjected to ligation of the left hepatic bile duct, the phosphorylative activity of the liver mitochondria in the ligated lobe decreased to about 50% of controls 48 h after the ligation, while that in the nonligated lobe increased 2-fold. These rabbits continued to live. On the other hand, in the rabbits subjected to ligation of the common bile duct, the mitochondrial phosphorylative activity increased slightly at 3 h after the ligation but fell rapidly thereafter. The mitochondrial free NAD+ to NADH ratios and hepatic energy charge (ATP+1/2ADP/ATP+ADP+AMP) markedly decreased after the ligation. Most of these rabbits did not survive over 2 days. These results indicate that the metabolic disturbances in the jaundiced liver are due to an impairment of the oxidative phosphorylation. In addition, it is suggested that the maintenance of the energy balance by enhanced mitochondrial phosphorylative activity is readily impaired in the jaundiced liver.

NADH-specific dihydropteridine reductase (EC 1.6.99.7) has been purified from human erythrocytes in essentially homogeneous form. The molecular weight of the enzyme was estimated to be 46,000 by Sephadex G-100 gel filtration. The enzyme reacted with antiserum against NADH-specific dihydropteridine reductase from bovine liver and formed a single immunoprecipitin line in the Ouchterlony double-diffusion system. This precipitin line completely fused with that formed between the human liver enzyme and the antiserum. With use of 5,6,7,8-tetrahydro-6-methylpterin, Km values of the erythrocyte enzyme for NADH and NADPH were determined to be 0.94 and 47 mumol/l, respectively. Vmax values were 58.7 mumol/min/mg with NADH and 6.41 mumol/min/mg with NADPH. The average activity of NADH-specific dihydropteridine reductase of 9 human blood samples from healthy males (20-25 years old) was calculated to be approximately 600 mU/g of hemoglobin, 1.8 mU per 20 microliters of blood, or 1.9 mU per 10(8) erythrocytes.

In this investigation, oral Kanamycin was administered to young male rabbits for 3 days, the control group receiving only water. Partial hepatectomy was then performed in all of them, and mitochondrial activity, energy charge and portal vein ammonia were measured at 12 and 24 h postoperatively. It was found that mitochondria from oral Kanamycin-treated rabbits have higher respiratory control ratios, higher state 3 respiration and higher P/O ratios than the control group, suggesting a further enhancement of phosphorylative activity. Energy charge in the Kanamycin-treated group showed less acute changes suggesting a positive balance in the energy status of those livers. The ammonia level here was much lower than in the control group. These findings suggest that the removal or the decrease of the overloading effect of ammonia resulting from bacterial activities in the intestine, has a beneficial effect on the healing process in liver surgery.

In 25% hepatectomized rabbits, the mitochondrial phosphorylative activity and the energy charge [(ATP + 1/2 ADP)/(ATP + ADP + AMP)] levels of the remnant liver remained unchanged after hepatectomy. In 70% hepatectomized rabbits, the energy charge levels of the remnant liver decreased maximally 24 h after hepatectomy (p less than 0.001), simultaneously with a marked enhancement of mitochondrial phosphorylative activity (P less than 0.001). Afterward, both returned to normal levels within 4 days. In 93% hepatectomized rabbits, the energy charge levels fell rapidly, with an inhibition of mitochondrial phosphorylative activity, within 6 h after hepatectomy. When a 24-hour 70% hepatectomized rabbit was cross-circulated with a normal rabbit, the energy charge in the liver of the normal partner decreased to 0.732 at 30 min after the blood exchange and then returned to 0.855 while that in the remnant liver of the hepatectomized partner increased from 0.767 to 0.820 (p less than 0.001). Also, the energy charge values of a 24-hour 70% hepatectomized rabbit were increased by the extracorporeal normal liver perfusion. It is suggested that a decrease in the energy charge of the remnant liver is due, at least partly, to the accumulation of the substances consuming hepatic energy (hepatodepressant factors) in the systemic blood of hepatectomized rabbits.

To clarify the effects of partial ischemia and reflow on the mitochondrial metabolism of the rat liver, the afferent vessels supplying the left lateral and left half of medial lobes were occluded and then reperfused after given time periods of ischemia (30, 60, 90 and 120 min, groups A, B, C and D, respectively). Samplings were taken at 0, 10 and 60 min after reperfusion. The energy charge levels of ischemic lobes decreased rapidly from 0.85 +/- 0.01 in the sham group to 0.38 +/- 0.11, 0.35 +/- 0.07 and 0.34 +/- 0.06 in groups B, C and D, respectively. The phosphorylative activities of mitochondria isolated from ischemic lobes decreased gradually along with the time of ischemia. The reversal of mitochondrial function and energy charge levels following reperfusion was noted in groups A and B. In nonischemic lobes, the phosphorylation rate (nmol ATP/mg/min) increased from 90 +/- 6 in sham group to 125 +/- 12 and 130 +/- 9; 131 +/- 5 and 130 +/- 6; 123 +/- 6 and 122 +/- 17, and 138 +/- 6 and 138 +/- 13 at 10 and 60 min after reflow in groups A, B, C and D, respectively (p less than 0.05). The energy charge level of nonischemic lobes decreased from 0.85 +/- 0.01 of sham group to 0.80 +/- 0.03 in group D (p less than 0.05). From these results, it is concluded that the transitional zone for the reversal of mitochondria function and energy metabolism following prolonged liver ischemia appears at around 60 min.(ABSTRACT TRUNCATED AT 250 WORDS)

20 hepatectomized patients, both cirrhotic and noncirrhotic, were studied for the postoperative changes in the arterial blood ketone body ratio and standard liver function tests. It was found that serum enzyme activities, total protein, albumin, total bilirubin, total cholesterol and prothrombin time were less reliable direct indicators of the onset of postoperative hepatic insufficiency than the arterial blood ketone body ratio, which reflects the oxidoreduction state of liver mitochondria. The hepatectomized patients were classified into three groups according to the postoperative changes in the blood ketone body ratio. In the 8 patients of group A, the blood ketone body ratio did not decrease to below 0.7. In the 7 patients of group B, the blood ketone body ratio decreased transiently to 0.4-0.7 and then increased to over 0.7. In the 5 patients of group C, the blood ketone body ratio decreased irreversibly to below 0.4. There were few postoperative complications in group A. In group B, while a variety of complications occurred, they were ameliorated concomitantly with the restoration of the blood ketone body ratio. All group C patients died of multiple organ failure. It is proposed that the arterial blood ketone body ratio is a new and practical approach which enables more effective postoperative management by the accurate evaluation of hepatic functional reserve after hepatectomy.

The effects of postoperative infusion of a hypertonic glucose solution on the blood glucose level, blood ketone body ratio (acetoacetate/beta-hydroxybutyrate), and plasma alanine and proline levels were studied in 70% hepatectomized rabbits (group A) and in rabbits 70% hepatectomized and, in addition, subjected to bile duct obstruction at 12 h after hepatectomy (group B). Glucose infusion was started at the end of hepatectomy and continued for 20 h. The blood glucose level in group A remained at approximately 300 mg/dl throughout the study; however, it reached 789 mg/dl in group B at 20 h. The blood ketone body ratio, which reflects hepatic mitochondrial redox potential, decreased from 0.90 +/- 0.09 in untreated rabbits to 0.38 +/- 0.05 in group A, and to 0.19 +/- 0.03 in group B at 20 h. As the blood ketone body ratio decreased, plasma proline and alanine levels increased rapidly (proline, r = -0.601, p less than 0.02; alanine, r = -0.640, p less than 0.001). In addition, the blood ketone body ratio was positively correlated with the hepatic energy charge level [(ATP + 0.5 ADP)/(ATP + ADP + AMP)] (r = 0.57, p less than 0.001). It is suggested that the entry of glucose and amino acids into the Krebs cycle is inhibited as the blood ketone body ratio decreases, and under such conditions the infused glucose tends to accumulate, resulting in severe hyperglycemia.

3 patients with chronic myelomonocytic leukemia (CMMoL) were investigated using a culture technique in vitro for granulocyte and monocyte/macrophage precursors (GM-CFC). Time course studies revealed that the growth of colonies and clusters was delayed in CMMoL as compared with that in normal control bone marrow. In 1 of 3 cases of CMMoL, the number of GM colonies increased after removal of phagocytes. Chloroacetate esterase and nonspecific esterase activities in the colonies and clusters were examined by means of our improved cytochemical technique. When we classified colonies and clusters into granulocytic, monocyte/macrophagic, myelomonocytic and mixed type, the incidence of "myelomonocytic' colonies and clusters was significantly higher in two cases of CMMoL than in the normal controls. On the basis of these results, the characteristics of abnormal hemopoietic regulatory mechanisms were discussed.

In order to study the changes in erythroid precursor cells in erythroleukemia, bone marrow cells from 4 patients were cultured for erythroid colony-forming units (CFU-e). The bone marrow from 3 of the patients showed an excess of ringed sideroblasts, and the incidence of CFU-e was very low. The one patient without ringed sideroblasts showed abundant erythroid colony formation. Erythroid colonies of more than eight erythroblasts consisted of normal-appearing erythroblast, while ringed sideroblasts were observed in scattered erythroblasts or in small clusters. The number of granulocyte macrophage colony-forming units (CFU-GM) was markedly lower than normal in all 4 cases. In 2 cases investigated, blast colonies were formed from bone marrow cells under the stimulation of phytohemagglutinin/leukocyte-conditioned medium. These results show that the differentiation of hemopoietic stem cells to erythroid as well as to myeloid cells is affected in erythroleukemia.

Glomerular localization of Hageman factor and fibrin-related antigen (FRA) was examined in 31 cases of IgA nephropathy by immunofluorescent techniques. Hageman factor was observed in 21 cases (68%) and FRA in 24 cases (77%). It is suggested that blood coagulation occurs and fibrin is formed in the glomerulus of IgA nephropathy.

Serum secretory IgA was measured to elucidate the significance of secretory IgA in patients with IgA nephropathy. The levels of serum secretory IgA and IgA were, respectively, 6.8 +/- 3.5 micrograms/ml and 231.0 +/- 69.2 mg/dl in the controls and 11.8 +/- 3.2 micrograms/ml and 385.3 +/- 78.7 mg/dl in the patients. The levels of serum secretory IgA and IgA in the patients were significantly higher than those in controls (p less than 0.01). Elevated serum secretory IgA may reflect the excessive state of the IgA-secreting system in IgA nephropathy patients.