Living-related liver transplantation in Diego blood group disparity: a case report.

To date, only limited cases of Diego blood group disparity in liver transplantation have been reported, and no cases with a long-term clinical course have been documented. Herein, we report a case of Diego blood group disparity in liver transplantation with details of long-term follow-up. The recipient was a 47-year-old woman with primary biliary cirrhosis; her 18-year-old daughter was the donor. Both recipient and donor were of blood type O according to the ABO blood group system. Preoperative serological tests showed the presence of antibodies against the Di(a) antigen only in the recipient, and not in the donor. Thus, the Diego phenotype was Di(a+) in the donor and Di(a-) in the recipient. Living-related liver transplantation was performed in July 2009. Immediate graft function was obtained, and no signs of humoral or cellular rejection were observed during the postoperative period. Further, anti-Di(a) antibodies were not detected throughout the postoperative course. The patient is alive and shows no signs of humoral rejection 34 months after liver transplantation. Liver transplantation has been performed successfully in cases of Diego blood group disparity.

No improvement in long-term liver transplant graft survival in the last decade: an analysis of the UNOS data.

We analyzed change in outcomes during two successive 5-year periods (period I = 1992-1996 vs period II = 1997-2002) among 35 186 deceased adult liver transplant recipients reported to the United Network for Organ Sharing (UNOS) Registry. The 5-year graft survival was 67.4% in the first period and 67.5% in the second, though the 1-year survival had improved from 81.0 to 83.5%. Comparison of blended survival rates during the two study periods showed decreased long-term graft survival in period II, explicable by an increased number of hepatitis C virus cirrhosis (HCV) patients and an increase in patients with HCV antibodies (HCVab) during this later period. Analysis wherein these patients with HCV were excluded revealed the same long-term graft survival during both periods. Non-HCV patients who had HCVab also had worse 5-year graft survival. We conclude that hepatitis C prevented improved outcomes during period II and that improved, more effective, treatment for hepatitis C virus would have great positive impact on overall survival of liver transplant recipients.

Efficient and cancer-selective gene transfer to hepatocellular carcinoma in a rat using adenovirus vector with iodized oil esters.

Gene therapy for cancer requires efficient, selective gene transfer to cancer cells. In gene therapy for hepatocellular carcinoma (HCC), gene transfer is efficient for small tumors, but not for large tumors. The delivery of anticancer agents and of iodized oil esters as embolic agents through tumor-feeding arteries is known as transarterial embolization. We speculate that genes may be efficiently and selectively transferred for HCC using iodized oil esters because these esters may remain together with a genetic vector within HCC selectively. Hence, we have studied the effect of iodized oil esters on adenovirus vector-mediated gene transfer for HCC in vivo. A rat model of HCC induced with diethylnitrosamine and phenobarbital was injected with either AxCALacZ, which expresses the beta-galactosidase of Escherichia coli, or AxCALacZ and iodized oil esters into the hepatic artery. Histological comparisons revealed that the beta-galactosidase expression in the rats with HCC injected with AxCALacZ and iodized oil esters was greater (P<.0001) in small tumors (P=.0046) and large tumors (P=.0023), and more selective (P=.0229) than in only AxCALacZ-injected rats. These results suggest that iodized oil esters are injected into hepatic artery together with adenovirus vector, and that genes may be efficiently and cancer-selectively transferred to HCC.

Efficiency of adenovirus-mediated gene transfer into hepatocytes by liver asanguineous perfusion method.

Efficient targeted gene delivery is essential for successful gene therapy. In this study, we examined the liver asanguineous perfusion method (LAP) for adenovirus-mediated gene transfer to the liver from the standpoints of efficiency, tissue-specificity and safety. The adenoviral vector containing the E. coli LacZ gene driven by the CAG promoter was delivered to the livers of rats by LAP. This method involves selective in situ perfusion, with the liver isolated by clamping of the afferent and efferent blood vessels to prevent adenoviral vector dissemination and genetic modification of nonhepatic organs. We demonstrated that gene transfer to the liver by LAP was not uniform, but more efficient than by intravenous (i.v.) or intraportal (i.p.) infusion, and caused no obvious liver damage or high mortality. As determined by specific histochemical staining and polymerase chain reaction, the amount of vector DNA transferred to the nonhepatic organs by LAP was significantly less than that transferred by the other two methods. Our data suggest that LAP is clearly superior to i.v. or i.p. infusion in terms of efficiency, specificity and safety of gene delivery to the liver. Further reduction in surgical risk is needed for the clinical application of gene therapy.

Inflamed site-specific gene delivery using bone marrow-derived CD11b+CD18+ vehicle cells in mice.

We report a novel technique that may allow site-specific gene delivery into inflamed tissues. Bone marrow cells from DBA/2 mice were incubated for 7 days in L-929 cell-conditioned medium containing elements that favor the development of mononuclear cells, such as colony-stimulating factors. Flow cytometric analysis revealed that 99.1 +/- 0.9% of the subcloned cells were positive for CD11b and CD18, both of which are ligands of the intercellular adhesion molecule 1 (ICAM-1). These vehicle cells were labeled with a fluorescent lipophilic probe and returned intravenously to the DBA/2 mice. The mice then received, for 1 week, intraperitoneal injections of either lipopolysaccharide (LPS) to enhance ICAM-1 expression in the glomerulus, or saline as a control. In the LPS-treated mice, labeled vehicle cells were detected within the glomerulus cross-section (gcs) 24 hr after the first injection (0.73 +/- 0.10/gcs). The number of labeled vehicle cells within the glomerulus gradually increased for 1 week (1.47 +/- 0.19/gcs) and decreased after discontinuation of the LPS injections. However, in the saline-treated control group, only a negligible number of vehicle cells could be detected in the glomerulus (0.05 +/- 0.03/gcs). A second administration of LPS 4 weeks after injection of the vehicle cells was also able to promote accumulation in the glomerulus. Furthermore, immunohistochemical analysis revealed that the kinetics of the vehicle cell recruitment into the glomerulus corresponded to the level of ICAM-1 expression. On the assumption that the LPS-induced ICAM-1 expression may regulate the site and timing of the delivery of vehicle cells into the glomerulus, vehicle cells were transduced with human glucocerebrosidase (GC) gene, using an adenovirus vector, and reintroduced into the mice. The basal expression of GC gene in the isolated glomeruli of vehicle cell-treated mice rose by 1.7-fold compared with endogenous activity, whereas the GC activity was enhanced 3.2-fold by LPS treatment. Polymerase chain reaction designed to detect human GC-specific sequence revealed that isolated glomeruli of vehicle cell-treated mice contained exclusively the vehicle cell-oriented GC. This indicates that vehicle cells can be used to carry a certain gene to a specific inflamed site. Injection of vehicle cells, with or without LPS, had small effect on urinary protein excretion or serum creatinine levels. These findings suggest that our novel method allows site-specific gene delivery into inflamed glomeruli through interaction of adhesion molecules.

Cholesterol reduces and corticosteroids enhance the toxicity of vitamin D in rats.

The effect of a high-cholesterol (CHOL) diet and corticosteroids on the toxicity of vitamin D2 (VD2) in rats was studied. VD2 was administered orally at the dosage of 5-60 x 10(4) IU/kg, once daily for 4 days. Animals fed CHOL showed a decrease in mortality due to VD2 treatment. Dietary CHOL inhibited toxic responses such as a diminished growth rate following anorexia, elevated serum calcium level and calcium deposition in tissues, which were produced by a sublethal dose of VD2 (20 x 10(4) IU/kg, once daily for 4 days). Animals pretreated with the high-CHOL diet from 2 weeks before the first VD2 administration showed much more symptomatic relief than those given this diet after the first VD2 administration. On the other hand, dexamethasone (DEX) as well as corticosterone remarkably increased the mortality due to VD2. The degree of VD2 toxicity, enhanced by DEX, was correlated with the degree of hypercalcemia and tissue calcification. Therefore, the inhibitory effect of CHOL is not likely to be due to activation of the CHOL-corticosterone system in the adrenal gland.

[Effect of dietary fats on serum and aortic lipid levels of mice fed a high-cholesterol diet: a distinct correlation between linoleic acid intake and the lipid changes].

The effects of dietary linoleic acid on serum lipids, lipid peroxides and aortic cholesterol were studied in mice fed a purified diet enriched with 5% cholesterol for a period of 14 weeks. The diet was supplemented with 10% coconut oil (Group I), lard (Group II), corn oil (Group III) or linoleic acid (Group IV) to give various levels of linoleic acid. After 4 to 12 weeks, the increment of serum total cholesterol was retained in the following order: group IV greater than III greater than II greater than I, which was the same order as the linoleic acid content in the diet. At week 14, the levels of serum free and esterified cholesterol, HDL-cholesterol, triglycerides and phospholipids were highest in group IV and lowest in group I. The serum lipid peroxide level was higher in the order of group IV greater than III greater than II greater than I. The ester ratio of cholesterol, the atherogenic index and LCAT activity were not significantly different among the four groups. Gallstone formation was markedly observed with higher dietary linoleic acid intake. Aortic cholesterol levels also increased in the same order as the dietary linoleic acid level: group IV greater than III greater than II greater than I. There were significant positive correlations between the aortic cholesterol level and all the serum lipid levels, and also the lipid peroxide level. All these findings indicate that under hypercholesterolemic conditions, excess dietary linoleic acid can increase serum lipids and lipid peroxide levels, resulting in lipid deposition in the aorta.(ABSTRACT TRUNCATED AT 250 WORDS)

Suppression of adjuvant arthritis in rats by cholesterol.

Dietary cholesterol suppressed adjuvant arthritis, a chronic inflammatory disease, in rats, but did not significantly affect carrageenin edema, an acute inflammation. When rats were fed a high-cholesterol diet beginning 10 days before injection of adjuvant, the development of the adjuvant-induced arthritis was greatly suppressed. Cholesterol feeding prevented hypertrophy of the adrenal gland in arthritic rats, but had little influence on the serum corticosterone level. A significant positive correlation was observed between the adrenal weight and the severity of the arthritis. These findings suggest that the effect of cholesterol feeding is not due to increased adrenal sterol synthesis. Dietary cholesterol also prevented hypertrophy of the spleen, but had no effect on atrophy of the thymus in adjuvant-treated rats. Cholesterol-fed rats showed a significant decrease in the serum lipid peroxide level and a significant increase in the serum copper level. Adjuvant treatment not only enhanced hypercholesterolemia produced by cholesterol feeding, but also the level of free cholesterol in serum. These results suggest that dietary cholesterol may exert some effect on the immune response through changes in spleen and liver functions.

Lipid deposition in the aorta of adjuvant arthritic rats with hypercholesterolemia.

This study offers findings which should aid in the development of a convenient animal model of atherosclerosis. Inbred Fisher strain rats were fed an atherogenic diet containing 1.5% cholesterol and 0.5% cholic acid and given a single subcutaneous injection of adjuvant (Mycobacterium butyricum) into the base of the tail. The animals were maintained for 8 weeks. Rats given the atherogenic diet showed markedly increased serum cholesterol levels, and all of those given the adjuvant injection developed severe polyarthritis. Cholesterol feeding tended to delay the onset of arthritis and remarkably suppressed the inflammatory response, particularly in the early stage of development. This may have been due to the lowered lipid peroxide levels in the serum of rats fed the atherogenic diet. Adjuvant arthritis together with cholesterol feeding markedly increased the cholesterol content of the aorta, whereas either treatment alone had little effect. The amounts of the connective tissue components and minerals in the aorta were not changed by both treatments. These results show that early atherosclerosis could be produced under the conditions used and that chronic inflammation and hypercholesterolemia are principal factors in the pathogenesis.

Hyperlipidemic effects of adriamycin in rats.

Adriamycin, an anticancer drug, caused dramatic increases in the serum lipid levels of rats fed a high-cholesterol diet. Male Lewis inbred rats were fed a basal or 1.5% cholesterol diet containing 0.5% cholic acid for 8 weeks. The rats were injected with adriamycin in doses of 1.5 mg/kg body weight, twice a week, and 6.0 mg/kg body weight, every other week. The serum lipid peroxide level gradually rose in adriamycin-treated rats, reaching a four-fold level at the end of the experiment. Cholesterol feeding, however, had a lowering effect on the lipid peroxide level. Adriamycin treatment or cholesterol feeding moderately elevated serum lipid levels, but their combination exerted a synergistic effect. In rats injected with a large dose of adriamycin and fed a high-cholesterol diet, the serum cholesterol, triglyceride and phospholipid levels strikingly increased by approx. 2000, 1500 and 1300 mg/100 ml, respectively. However, the ester ratio of cholesterol remained almost constant. Furthermore, serum GOT, GPT and ALP activities were only slightly different from the control values. Adriamycin treatment produced severe hypoalbuminemia. Ascites was also observed in rats given a large dose of adriamycin. The present findings indicate that the hyperlipidemia we observed may basically result from adriamycin-induced nephrosis and can be markedly enhanced when rats are fed a high-cholesterol diet. In spite of remarkably high levels of serum lipids and lipid peroxides, the aortic cholesterol level increased only slightly.