bluuebunny/crossref_metadata_2025 huggingface dataset:
Local RAD50 gene delivery induces regression of preformed porcine coronary in‐stent neointimal hyperplasia
Authors: Ahn Young Keun , Kook Hyun , Jeong Myung Ho , Ahn Kyu Youn , Cho Jeong Gwan , Park Jong Chun , Kang Jung Chaee , Kim Kyung Keun
Published: January 2004

Background

Recently, we observed that overexpression of human RAD50 (hRAD50) induced p21‐dependent cytotoxicity in various cultured cells, and rat and mouse tumor models. This study investigated the characteristics of endothelial cell (EC) death by hRAD50 and the potential utility of hRAD50 in the development of gene therapies for vascular restenosis.

Methods

We studied the effects of transient hRAD50 gene transfer using nonliposomal lipid on the survival of primary cultured human coronary arterial EC and smooth muscle cells (SMC). Palmaz‐Schatz stents were deployed in two epicardial coronary arteries in each pig (n = 10). Two weeks later, the patency of the stented arteries was documented by coronary angiography, and the hRAD50 construct or empty vector mixed with lipid was delivered to one of the stented arteries in each pig using a Dispatch catheter. Coronary angiography was repeated 2 weeks after gene delivery and histological examination was performed.

Results

Lipid‐mediated hRAD50 gene transfer resulted in the death of EC and SMC. It also increased endothelial nitric oxide synthase (eNOS) expression and nitrite production as well as p21 expression. Pretreatment with NOS and pan‐caspase inhibitors completely prevented EC death by hRAD50. In the hRAD50‐delivered arteries, the percentage of diameter stenosis, neointimal area, and pathologic area of stenosis were significantly smaller than in the control arteries. eNOS expression increased in the hRAD50‐delivered arteries. Systemic hematologic and chemical values were not affected by gene delivery.

Conclusions

Significant regression of preformed in‐stent neointimal hyperplasia was induced by local hRAD50 gene delivery to stented porcine coronary arteries without apparent systemic toxicity. Copyright © 2003 John Wiley & Sons, Ltd.
In vivo expansion of gene‐modified hematopoietic cells by a novel selective amplifier gene utilizing the erythropoietin receptor as a molecular switch
Authors: Nagashima Takeyuki , Ueda Yasuji , Hanazono Yutaka , Kume Akihiro , Shibata Hiroaki , Ageyama Naohide , Terao Keiji , Ozawa Keiya , Hasegawa Mamoru
Published: January 2004

Background

In vivo expansion of gene‐modified cells would be a promising approach in the field of hematopoietic stem cell gene therapy. To this end, we previously developed a selective amplifier gene (SAG), a chimeric gene encoding the granulocyte colony‐stimulating factor (G‐CSF) receptor (GCR), as a growth‐signal generator and the hormone‐binding domain of the steroid receptor as a molecular switch. We have already reported that hematopoietic cells retrovirally transduced with the SAG can be expanded in a steroid‐dependent manner in vitro and in vivo in mice and nonhuman primates. In this study, we have developed a new‐generation SAG, in which the erythropoietin (EPO) receptor (EPOR) is utilized instead of the steroid receptor as a molecular switch.

Methods

Two EPO‐driven SAGs were constructed, EPORGCR and EPORMpl, containing the GCR and c‐Mpl as a signal generator, respectively. First, to compare the steroid‐driven and EPO‐driven SAGs, Ba/F3 cells were transduced with these SAGs. Next, to examine whether GCR or c‐Mpl is the more suitable signal generator of the EPO‐driven SAG, human cord blood CD34+ cells were transduced with the two EPO‐driven SAGs (EPORMpl and EPORGCR). Finally, we examined the in vivo efficacy of EPORMpl in mice. Irradiated mice were transplanted with EPORMpl‐transduced bone marrow cells followed by administration of EPO.

Results

The EPO‐driven SAGs were shown to induce more rapid and potent proliferation of Ba/F3 cells than the steroid‐driven SAGs. The EPORMpl induced more efficient EPO‐dependent proliferation of the human cord blood CD34+ cells than the EPORGCR in terms of total CD34+ cell, c‐Kit+ cell, and clonogenic progenitor cell (CFU‐C) numbers. In the transplanted mice the transduced peripheral blood cells significantly increased in response to EPO.

Conclusions

The new‐generation SAGs, especially EPORMpl, are able to efficiently confer an EPO‐dependent growth advantage on transduced hematopoietic cells in vitro and in vivo in mice. Copyright © 2003 John Wiley & Sons, Ltd.
Partial correction of the α‐galactosidase A deficiency and reduction of glycolipid storage in Fabry mice using synthetic vectors
Authors: Przybylska Malgorzata , Wu I‐Huan , Zhao Hongmei , Ziegler Robin J. , Tousignant Jennifer D. , Desnick Robert J. , Scheule Ronald K. , Cheng Seng H. , Yew Nelson S.
Published: January 2004

Background

Fabry disease is a recessive, X‐linked disorder caused by a deficiency of the lysosomal enzyme α‐galactosidase A, leading to an accumulation of the glycosphingolipid globotriaosylceramide (GL‐3) in most tissues of the body. The goal of this study was to determine if systemic delivery of a nonviral vector could correct the enzyme deficiency and reduce the levels of GL‐3 in different tissues of a transgenic knockout mouse model of the disease.

Methods

Cationic lipid was complexed with a CpG‐depleted plasmid DNA vector and then injected intravenously into Fabry mice. The levels of α‐galactosidase A and GL‐3 in different tissues were assayed at various time points after injection.

Results

Expression of α‐galactosidase A was detected in the different tissues of Fabry mice for up to 3 months after complex administration, but resulted in minimal reductions in GL‐3 levels. However, the use of the anti‐inflammatory drug dexamethasone and multiple dosing increased α‐galactosidase A expression and resulted in significant reductions of GL‐3 in all the organs with the exception of the kidney. In addition, injecting complex into young Fabry mice partially prevented the normal accumulation of GL‐3 in the heart, lung, and liver.

Conclusions

Systemic delivery of a cationic lipid–pDNA complex partially corrected the enzyme deficiency and reduced glycolipid storage in a mouse model of Fabry disease. The results are one of the few demonstrations of long‐term efficacy in a genetic disease model using nonviral vectors. However, substantial improvements in expression, especially in critical organs such as the kidney, are required before these vectors can become a viable approach to treat Fabry disease and other lysosomal storage disorders. Copyright © 2003 John Wiley & Sons, Ltd.
Factors influencing immune response after in vivo retrovirus‐mediated gene transfer to the liver
Authors: Podevin Guillaume , Otta Edson , Nguyen Jean Michel , Pichard Virginie , Aubert Dominique , Moullier Philippe , Ferry Nicolas
Published: January 2004

Background

Highly efficient retrovirus‐mediated gene transfer into hepatocytes in vivo triggers an immune response directed against transduced hepatocytes. This effect may be due either to spreading of retroviral vectors in the blood stream with subsequent infection of antigen presenting cells (APCs) or to cross‐presentation of the transgene product present as a contaminant in the viral stock. In order to decrease immune response, we evaluated the effect of asanguineous perfusion of the liver as well as purification of the viral stock on long‐term transduction of hepatocytes using the nls‐lacZ marker gene.

Methods

Animals were divided in four groups. In group 1, the viral supernatant was perfused in the regenerating liver after complete vascular exclusion of the organ. In group 2, using the same strategy, animals received retroviral supernatant that was passed through a β‐galactosidase affinity column to reduce β‐galactosidase contamination. In two control groups (respectively groups 3 and 4) the corresponding viral supernatants were delivered via peripheral injection.

Results

In group 1, 23.1% of animals had no immune response 2 months after gene delivery vs. 33.4% in group 2, 4.3% in control group 3, and 0% in control group 4. Statistical analysis of the results demonstrated that only the difference between groups 2 and 3 was statistically significant. This indicated that both asanguineous perfusion together with passage through an affinity column were required to decrease significantly immune response.

Conclusions

Our present results suggest that both supernatant contamination and viral spreading contribute to immune response after retrovirus‐mediated gene delivery to the liver. Copyright © 2003 John Wiley & Sons, Ltd.
In situ single cell observation by fluorescence resonance energy transfer reveals fast intra‐cytoplasmic delivery and easy release of plasmid DNA complexed with linear polyethylenimine
Authors: Itaka Keiji , Harada Atsushi , Yamasaki Yuichi , Nakamura Kozo , Kawaguchi Hiroshi , Kataoka Kazunori
Published: January 2004

Background

The investigation into the intracellular mechanisms for gene expression has acquired great impetus for the improvement of the transfection efficiency by a non‐viral gene delivery system.

Methods

Intracellular trafficking as well as release of plasmid DNA (pDNA) complexed with polycations, including linear and branched polyethylenimine (LPEI, BPEI) and poly(L‐lysine) (PLL), were explored under confocal microscopy using fluorescence resonance energy transfer (FRET) between a pair of donor–acceptor fluorescent dyes (fluorescein and Cy3) tagged on a single pDNA molecule.

Results

pDNA complexed with LPEI underwent a rapid escape from the endosomes, spreading uniformly into the cytoplasm with a substantial decrease in FRET efficiency due to the disintegration of LPEI/pDNA polyplex structure. pDNA complexed with BPEI also achieved a rapid escape from the endosomes. Nevertheless, the pDNA retained high FRET efficiency even after 24 h, indicating an appreciable stability of the BPEI/pDNA polyplex to keep pDNA in a condensed state. In the PLL/pDNA polyplexes, neither endosome escape nor pDNA decondensation was observed. These intracellular characteristics of polyplexes showed a clear correlation to their gene transfection efficiency: The LPEI/pDNA revealed a considerably higher and faster gene expression compared with BPEI/pDNA. Atomic force microscopy revealed that BPEI induced more effective condensation of pDNA than LPEI, being consistent with restricted cytoplasmic release of complexed pDNA.

Conclusion

Fast endosomal escape and subsequent smooth disintegration of LPEI/pDNA in the cytoplasm are likely to be determining factors for the excellent transfection efficiency of this polyplex system. These properties may be particularly beneficial to achieve appreciably high gene expression in a prompt manner. Copyright © 2003 John Wiley & Sons, Ltd.
Use of the norepinephrine transporter as a reporter gene for non‐invasive imaging of genetically modified cells
Authors: Anton Martina , Wagner Bettina , Haubner Roland , Bodenstein Claudia , Essien Bryan E. , Bönisch Heinz , Schwaiger Markus , Gansbacher Bernd , Weber Wolfgang A.
Published: January 2004

Background

The norepinephrine transporter (NET) is a high‐affinity transporter for catecholamines. Its expression is almost exclusively restricted to the sympathetic nervous system. In this study we evaluated whether the NET can be used as a reporter gene for non‐invasive imaging of genetically modified cells with radiolabeled probes.

Methods

Human A431, HT1080 and murine CMS‐5 cells were retrovirally transduced with bovine NET cDNA. Transduced and parental cells were incubated in vitro with [131I]meta‐iodobenzylguanidine ([131I]MIBG). The specificity of tracer uptake was determined by adding the NET inhibitor imipramine. Rat PC12 cells served as positive controls. Parental and A431NET cells were xenotransplanted into nude mice and tumor uptake of [123I]MIBG in vivo was determined after tracer administration.

Results

In vitro stably transduced cells showed a 66‐ to 120‐fold higher [131I]MIBG uptake than parental cells. Incubation with imipramine reduced [131I]MIBG uptake of transduced cells to the level found in parental cells. More than 70% of the initial radioactivity was retained in all transduced cell lines after 2 h incubation with tracer‐free medium. [131I]MIBG uptake in PC12 cells, which express the NET endogenously, was 20‐ to 28‐fold lower than in transduced cells. In vivo, A431NET tumors demonstrated a 33‐fold higher [123I]MIBG uptake than parental tumors. Gamma camera images 24 h after tracer injection showed no tracer uptake in parental A431 tumors, but clear images of A431NET tumors.

Conclusions

Transduction of tumor cells with NET cDNA causes highly specific uptake and significant retention of catecholamine analogs in vitro and in vivo. These characteristics make the NET suitable as a reporter gene for non‐invasive monitoring of gene transfer. Copyright © 2003 John Wiley & Sons, Ltd.
Gene‐directed enzyme prodrug therapy for prostate cancer in a mouse model that imitates the development of human disease
Authors: Martiniello‐Wilks Rosetta , Dane Allison , Voeks Dale J. , Jeyakumar Geetha , Mortensen Elin , Shaw Jan M. , Wang Xiao Yang , Both Gerald W. , Russell Pamela J.
Published: January 2004

Background

Gene‐directed enzyme prodrug therapy (GDEPT) based on the E. coli enzyme purine nucleoside phosphorylase (PNP) represents a new approach for treating slow growing tumours like prostate cancer (PCa). Expressed enzyme converts a systemically administered prodrug, fludarabine phosphate, to a toxic metabolite, 2‐fluoroadenine. Infected and neighbouring cells are killed by a bystander effect that results from the inhibition of DNA and RNA synthesis.

Methods

These studies were carried out using the transgenic adenocarcinoma of the prostate (TRAMP) model that mimics human PCa development and progression. Control TRAMP mice were injected intraprostatically with vector vehicle and thereafter intraperitoneally with saline or fludarabine phosphate (∼600 mg/m2/day) once daily for 5 consecutive days. Treated mice received a single intraprostatic injection containing 1010 particles of OAdV220, an ovine atadenovirus which expresses the E. coli PNP gene under the control of the Rous sarcoma virus promoter, followed by systemic fludarabine treatment. The weight of the genitourinary tract, seminal vesicles and the prostate as well as animal survival were monitored. Tumours were also analysed histologically.

Results

Preliminary studies showed that fludarabine alone caused no significant change in genitourinary (GU) tract weight in TRAMP mice. Animals injected with vector and prodrug showed a significant reduction (36–47%) in GU tract weight (ANOVA p = 0.0002) and a 35–50% reduction in seminal vesicle weight (ANOVA p = 0.0007). In particular, the target organ showed a significant 57% reduction in prostate weight (ANOVA p = 0.0007). PNP‐GDEPT mice also showed a survival advantage over control mice. Histological analysis suggested that the cancer progression was slowed in GDEPT‐treated animals.

Conclusion

A single course of GDEPT based on OAdV‐delivered PNP and fludarabine produced highly significant suppression of PCa progression in immune‐competent TRAMP mice. Copyright © 2004 John Wiley & Sons, Ltd.
Steric stabilization of poly(2‐(dimethylamino)ethyl methacrylate)‐based polyplexes mediates prolonged circulation and tumor targeting in mice
Authors: Verbaan F. J. , Oussoren C. , Snel C. J. , Crommelin D. J. A. , Hennink W. E. , Storm G.
Published: January 2004

Background

Efficient tumor targeting of polymeric gene transfer systems (polyplexes) represents a major challenge. To establish tumor targeting after intravenous (IV) administration, the circulation lifetime of these systems should be sufficiently long. Since naked polyplexes are rapidly eliminated from the circulation after IV adminstration, strategies have to be developed to improve their pharmacokinetics.

Methods

Complexes of plasmid DNA and poly(2‐(dimethylamino)ethyl methacrylate) (pDMAEMA)‐graft‐PEG or AB di‐block copolymers of pDMAEMA and PEG, as well as PEGylated complexes prepared via PEGylation of preformed complexes (postPEGylation), were evaluated for their physicochemical properties (size and charge) their interactions with blood constituents and transfection activity in vitro. The pharmacokinetics and biodistribution of PEG‐polyplexes were studied in mice after IV administration. The degree of accumulation in two subcutaneous (SC) mouse tumors after IV administration was evaluated for the system with the longest circulation time.

Results

It is shown that the surface charge of the pDMAEMA‐polyplexes was effectively shielded by two PEGylation methods (i.e. the use of pDMAEMA‐graft‐PEG polymers and postPEGylation). The shielding effect was the highest for the postPEGylation method with PEG20000, yielding polyplexes that hardly show interactions with blood components (i.e. albumin and erythrocytes) and show substantially prolonged circulation time in mice after IV administration. The superior colloidal stability and circulation kinetics of the postPEGylated polyplexes translated into tumor accumulation which amounted to about 3.5% of the injected dose per gram tumor tissue in a SC Neuro2A tumor model and to about 4.2% of the injected dose per gram tumor tissue in a SC C26 tumor model.

Conclusions

This study shows that postPEGylation of pDMAEMA‐based polyplexes is the most attractive method to prepare polyplexes with long circulating properties. Tumor targeting capacity after intravenous administration was demonstrated in two subcutaneous tumor models. Copyright © 2004 John Wiley & Sons, Ltd.
Developing a convenient large animal model for gene transfer to salivary glands in vivo
Authors: Li Jun , Zheng Chanyu , Zhang Xin , Liu Xiaoyong , Zhang Chengmei , Goldsmith Corinne M. , Baum Bruce J. , Wang Songlin
Published: January 2004

Background

Localized gene transfer to salivary glands has great potential for the treatment of salivary gland, systemic, and oral diseases. The minipig parotid gland, given its volume and morphological similarities to the human parotid gland, may be useful as a large animal model for pre‐clinical gene transfer experiments. The purpose of this study was to perform an initial assessment of the efficacy and safety of adenoviral‐vector‐mediated gene transfer to parotid glands of miniature pigs.

Methods

AdCMVluc, a recombinant type 5 adenoviral (rAd5) vector containing a luciferase reporter gene, was administered to miniature pig parotid glands by intraductal cannulation. Five regions of gland tissue were obtained to measure the distribution of luciferase activity. The effects of time, viral dose, infusate buffer volume, and gland anatomical region on transgene expression were determined. Detailed serum chemistry and hematological analyses were performed. In addition, AdCMVlacZ, a similar rAd5 vector encoding β‐galactosidase, was also delivered to determine the parotid gland cell types transduced.

Results

Luciferase assays indicated that gene transfer to miniature pig salivary glands could be readily accomplished using rAd5 vectors. Highest transgene expression was found in the center of glands, which was > posterior > inferior > anterior > superior tissue regions. Expression was maximal on day 2 and declined to background by day 14, and observed in both acinar and ductal cells. Several serum chemistry and hematology parameters were transiently changed following rAd5 administration.

Conclusions

Transgene expression by, and inflammatory response to, rAd5 vectors in minipig parotid glands are similar to results seen earlier in rodent studies. This suggests that results of salivary gland gene transfer from rodent studies can be extended to a larger animal model, and supports the value of using minipigs for pre‐clinical applications of gene transfer to these tissues. Published in 2004 by John Wiley & Sons, Ltd.
Local ex vivo gene therapy with bone marrow stromal cells expressing human BMP4 promotes endosteal bone formation in mice
Authors: Zhang Xiao S. , Linkhart Thomas A. , Chen Shin‐Tai , Peng Hairong , Wergedal Jon E. , Guttierez Genevieve G. , Sheng Matilda H.‐C. , William Lau K.‐H. , Baylink David J.
Published: January 2004

Background

Bone loss in osteoporosis is caused by an imbalance between resorption and formation on endosteal surfaces of trabecular and cortical bone. We investigated the feasibility of increasing endosteal bone formation in mice by ex vivo gene therapy with bone marrow stromal cells (MSCs) transduced with a MLV‐based retroviral vector to express human bone morphogenetic protein 4 (BMP4).

Methods

We assessed two approaches for administering transduced MSCs. β‐Galactosidase (β‐Gal) transduced C57BL/6J mouse MSCs were injected intravenously via tail vein or directly injected into the femoral bone marrow cavity of non‐marrow‐ablated syngenic recipient mice and bone marrow cavity engraftment was assessed. BMP4‐ or β‐Gal‐transduced cells were injected into the femoral bone marrow cavity and effects on bone were evaluated by X‐ray, peripheral quantitative computed tomography (pQCT), and histology.

Results

After tail‐vein injection less than 20% of recipient mice contained β‐Gal‐positive donor cells in femur, humerus or vertebra marrow cavities combined, and in these mice only 0.02–0.29% of injected cells were present in the bone marrow. In contrast, direct intramedullary injection was always successful and an average of 2% of injected cells were present in the injected femur marrow cavity 24 hours after injection. Numbers of donor cells decreased over the next 14 days. Intramedullary injection of BMP4‐transduced MSCs induced bone formation. Trabecular bone mineral density (BMD) determined by pQCT increased 20.5% at 14 days and total BMD increased 6.5% at 14 days and 10.4% at 56 days.

Conclusions

The present findings support the feasibility of using ex vivo MSC‐based retroviral gene therapy to induce relatively sustained new bone formation, with normal histological appearance, at endosteal bone sites. Copyright © 2004 John Wiley & Sons, Ltd.
A prospective cross-over study comparing the effect of mycophenolate versus azathioprine on allograft function and viral load in liver transplant recipients with recurrent chronic HCV infection
Authors: Zekry Amany , Gleeson Margret , Guney Seren , McCaughan Geoffrey W.
Published: January 2004

It has been suggested that Mycophenolate Mofetil (MMF) may have an antiviral effect in patients with recurrent HCV infection post-liver transplantation. We conducted a prospective cross-over study in liver transplant recipients with recurrent chronic HCV infection to examine whether substitution of azathioprine (AZA) with MMF would reduce HCV load and improve allograft function. Thirteen long standing HCV liver transplant recipients receiving AZA were enrolled in a 9-month prospective cross-over study. In the initial 3 months lead-in period, baseline viral loads and biochemistry were recorded. Following this, MMF was substituted for AZA at a dose of 1 gm twice/day for a period of 3 months after which patients were switched back to AZA and observed for a further 3 months. Viral loads, biochemical allograft function, and adverse effects were closely monitored during the study period. Thirteen patients (12 males and 1 female) were enrolled. The mean age was 54 (±8) years and the mean time from transplantation was 68 (±35) months. Baseline mean viral load was 0.74 × 106(±0.47 × 106) messenger RNA (mRNA) copies/ml. By the end of the MMF treatment period, the mean viral load increased to a level of 1.64 × 106 (±1.3 × 106) mRNA copies/ml (P = 0.026) compared to baseline. The increase in viral load however was not associated with an increase in ALT level. In a cohort of 13 HCV liver transplant recipients with recurrent chronic HCV infection, substitution of azathioprine with MMF did not lead to a decrease in viral load. (Liver Transpl 2004;10:52-57.)
Treatment of refractory cholestatic pruritus after liver transplantation with albumin dialysis
Authors: Bellmann Romuald , Graziadei Ivo W. , Feistritzer Clemens , Schwaighofer Hubert , Stellaard Frans , Sturm Ekkehard , Wiedermann Christian J. , Joannidis Michael
Published: January 2004

Albumin dialysis has been shown to improve the outcome in patients with cholestatic liver failure caused by chronic liver disease. This study reports 7 liver transplant recipients who were treated with albumin dialysis for intractable pruritus of different origin (ductopenic graft rejection, non-anastomotic strictures, and recurrence of hepatitis C). Treatment with histamine (H1) blockers, opioid antagonists, and cholestyramine had not been effective. The Molecular Adsorbent Recirculating System (MARS; Teraklin, Rostock, Germany) was used for albumin dialysis. All patients presented with numerous scratch marks, 6 of whom had a pronounced icterus. Six patients (86%) responded to 3 consecutive treatments with significant reduction of pruritus. The mean pruritus score, which was quantified by a visual analog scale (VAS), decreased from 9.7 ± 0.5 to 3.7 ± 0.8 (SD). The mean duration of 1 treatment was 15.6 hours. The procedure was well tolerated by all patients. The mean total serum bilirubin in patients who responded to therapy declined from 19.11 ± 16.96 mg/dL (SD) before MARS therapy to 9.24 ± 3.52 mg/dL after treatment. The mean serum concentration of 3α-hydroxy bile acids decreased from 192.67 ± 58.12 μmol/L (SD) to 42.33 ± 31.58 μmol/L (SD). Follow-up in 3 cases showed sustained improvement of pruritus lasting for more than 3 months. In 3 patients, however, pruritus relapsed. One patient, who showed severe pruritus, without relevant elevation of serum bile acids before treatment, did not respond to albumin dialysis. Our data indicate that MARS is an effective therapeutic option for patients with intractable cholestatic pruritus. (Liver Transpl 2004;10:107-114.)
Periportal edema and necrosis as diagnostic histological features of early humoral rejection in ABO-incompatible liver transplantation
Authors: Haga Hironori , Egawa Hiroto , Shirase Tomoyuki , Miyagawa Aya , Sakurai Takaki , Minamiguchi Sachiko , Yamabe Hirohiko , Manabe Toshiaki , Tanaka Koichi
Published: January 2004

Humoral rejection caused by antidonor blood group A/B antibodies is one of the most important obstacles for successful ABO-incompatible liver transplantation. However, no specific morphologic features of liver biopsies to distinguish humoral rejection from other conditions such as ischemia or sepsis have been satisfactorily documented. To histologically clarify the early changes in humoral rejection, we studied 41 cases of living donor ABO-incompatible liver transplantation whose allograft biopsies during the first episode of suspected acute rejection were available within the first postoperative month. Postoperative isohemagglutinin IgM titers were ×64 or more in 21 patients (51%; high-titer group) and less than ×64 in 20 cases (49%; low-titer group). In the high-titer group, elevation of postoperative titers ×64 or more occurred within postoperative days 5.7 ± 4.1 (range: 1-17). An increase in the incidence of cholangitis was observed in the high-titer group (90% vs. 30%, P < .0001), as well as poorer overall graft survival than in the low-titer group (38% vs. 70%, P < .05). Seven biopsies obtained from the high-titer group within 3 days after the onset of elevation of the antibody titers and one biopsy obtained at the peak of the antibody titers demonstrated periportal edema and necrosis, neither of which was found in the low-titer group. All grafts of these patients caused massive hepatocyte necrosis or severe biliary complications. In conclusion, a high morbidity rate of ABO-incompatible liver transplantation is associated with high postoperative levels of antibody titers. Periportal edema and necrosis observed during elevation of antibody titers can be regarded as histological indications of early changes in severe humoral rejection. (Liver Transpl 2004;10:16-27.)
A matched comparison study of medical and psychiatric complications and anesthesia and analgesia requirements in methadone-maintained liver transplant recipients
Authors: Weinrieb Robert M. , Barnett Rebecca , Lynch Kevin G. , DePiano Maria , Atanda Alfred , Olthoff Kim M.
Published: January 2004

Approximately 85% of patients receiving methadone maintenance therapy (MMT) for opiate dependence in the United States are infected with hepatitis C virus (HCV). MMT is significantly underrepresented in most liver transplant programs, but the number of patients receiving MMT is increasing and few data are available to guide treatment. We evaluated MMT in our program (27 pretransplant and 10 posttransplant cases) for medical and psychiatric complications and anesthesia and analgesia requirements. After transplant, 10 patients receiving MMT were compared with a matched control group of 19 patients who were not receiving MMT and not dependent on opiates. Fewer patients receiving MMT retained a spot on the transplant waiting list (65%) than patients not receiving MMT (80%); 30% of patients receiving MMT pretransplant used heroin, cocaine, or marijuana, and more than 25% were lost to follow-up. Liver disease according to mean Child-Turcotte-Pugh (CTP) score and transplant waiting times was similar between the 2 groups. Patients receiving MMT required significantly more intraoperative anesthesia and postoperative analgesia (mean fentanyl 3,175 μg/d, SD = 2,832; intravenous morphine 67.86 mg/d, SD = 38.84, respectively) compared with patients not receiving MMT (mean fentanyl 1,324 μg/d, SD = 1,122; intravenous morphine 12.17 mg/d, SD = 10.24, respectively). More patients receiving MMT had severe recurrent HCV infection (60%) and worse survival (60%) versus patients not receiving MMT (21% and 78.9%, respectively). Follow-up times did not differ between groups (MMT: mean 4.19 years, median 1.15 years, SD = 7.6; non-MMT: mean 2.68 years, median 2.19 years, SD = 1.73). Finally, patients receiving MMT required an average methadone dose increase of 60% from pretransplant to posttransplant. Postoperative analgesia guidelines are described. Posttransplant, 20% of patients receiving MMT used alcohol or illicit drugs. Data do not support withholding the provision of liver transplantation to patients receiving MMT, but larger, well-controlled studies are warranted. (Liver Transpl 2004;10:97-106.)
Hepatitis B virus genotype A and D and clinical outcomes of liver transplantation for HBV-related disease
Authors: Girlanda Raffaele , Mohsen Abdul H , Smith Heather , Sablon Erwin , Yuen Man-Fung , O’Grady John , Muiesan Paolo , Rela Mohamed , Heaton Nigel , Norris Suzanne
Published: January 2004

Hepatitis B virus (HBV) genotypes have been associated with specific patterns of disease and response to antiviral therapy. We investigated the effect of HBV genotype on HBV recurrence and mortality after liver transplantation (LT). Pretransplant sera of 45 hepatitis B surface antigen (HBsAg) positive adults were submitted for HBV genotyping by a reverse-phase hybridization line probe assay with genotype-specific probes. Data were correlated with clinical outcomes after transplantation. Genotype A (n =15), D (n = 13) and A/D (n = 12) accounted for 89% of all genotypes. Coinfection with two HBV genotypes was encountered in 14 (31.1%) patients. Eighteen patients (40 %) developed HBV recurrence at a median of 10 months posttransplant (range, 1-53) and 10 patients (22 %) died at a median of 24 months (range, 3-63). Genotype D patients were more likely to develop HBV recurrence or die compared with genotype A patients, although this did not reach statistical significance. Dual infection with genotype A/D resulted in mortality similar to that of genotype A but recurrence similar to that of genotype D. Active viral replication at time of transplantation was the only independent factor (P = 0.03) that predicted HBV recurrence. In conclusion, HBV genotype A and D did not have a significant impact on clinical outcomes of LT for HBV-related liver disease in patients of European origin. These data do not support routine HBV genotyping in liver transplantation. (Liver Transpl 2004;10:58-64.)
Native liver xanthogranulomatous cholangiopathy in primary sclerosing cholangitis: Impact on posttransplant outcome
Authors: Keaveny Andrew Paul , Gordon Fredric David , Goldar-Najafi Atoussa , Lewis William David , Pomfret Elizabeth Anne , Pomposelli James John , Jenkins Roger Lewin , Khettry Urmila
Published: January 2004

A retrospective analysis of 51 primary sclerosing cholangitis (PSC) patients who underwent liver transplant (LT) identified 16 with xanthogranulomatous cholangiopathy (XGC) at the native liver hilum. Pre-LT clinical and laboratory data and post-LT course and outcome of patients with XGC were compared with the 35 PSC patients without XGC. The XGC and non-XGC groups were similar with respect to age and laboratory data at the time of LT. Pre-LT cholecystectomy was performed in 44% versus 26% and biliary bypass procedure in 38% versus 26% of patients with and without XGC, respectively (P = NS). Peri-operative complications resulted in six (38%) deaths or retransplantation within 60 days of LT in the XGC group compared with 4 (11%) in the non-XGC group (P = .05). Patient survival at 60 and 100 days post-LT was better in the non-XGC group (P = .01). The causes of death or retransplantation within 60 days post-LT in the patients with XGC included primary nongraft function (1), uncontrolled bleeding (3), and sepsis (2), while in the non-XGC group these were uncontrolled bleeding (2), sepsis (1), and primary nongraft function (1). Mean graft survival ± SD was 1,081 ± 1,584 days in patients with XGC versus 2,149 ± 1,679 days in patients without XGC. The presence of XGC in the native liver hilum of PSC patients undergoing LT was associated with a higher rate of early post-LT mortality or retransplantation. In conclusion, no pre-LT clinical features or laboratory tests were identified that predicted the presence of XGC in PSC patients. (Liver Transpl 2004;10:115-122.)
Volume regeneration after right liver donation
Authors: Hata Shojiro , Sugawara Yasuhiko , Kishi Yoji , Niiya Takashi , Kaneko Junichi , Sano Keiji , Imamura Hiroshi , Kokudo Norihiro , Makuuchi Masatoshi
Published: January 2004

After right hepatectomy with the middle hepatic vein trunk for a graft, the venous outflow in segment IV is disturbed. There are limited data, however, regarding the effect of middle hepatic vein deprivation on liver regeneration or functional recovery. Living donors who underwent right hepatectomy with preservation of the middle hepatic vein (Group A, n = 58) and those deprived of the middle hepatic vein (Group B, n = 13) were reviewed. When the donor was under 50 years old and the remnant left liver was estimated to be more than 35% of the whole liver, right liver graft harvesting with the middle hepatic vein trunk was considered. Volume regeneration of segments I-III, segment IV, and overall liver volume was assessed at the third postoperative month using computed tomography. The regeneration rate of segment IV was significantly impaired in Group B donors compared with that in Group A donors (125% vs. 45%, P = 0.008). In contrast, the regeneration rate of segments I -III was significantly higher than that in Group A (208% vs. 263%, P = 0.004). There was no significant difference in the regeneration rate of the whole left liver or functional recovery between groups. Multivariate analysis revealed that the resection type (group) was a significant predictive factor for the regeneration rate of segments I-III and segment IV. When deprived of the middle hepatic vein, liver regeneration of segment IV was impaired but was compensated for by the regeneration of segments I-III. In conclusion, extended right hepatectomy can be safely performed with careful preoperative consideration using these criteria. (Liver Transpl 2004;10:65-70.)
Hepatectomy of living donors with a left-sided gallbladder and multiple combined anomalies for adult-to-adult living donor liver transplantation
Authors: Hwang Shin , Lee Sung-Gyu , Park Kwang-Min , Lee Young-Joo , Ahn Chul-Soo , Kim Ki-Hoon , Moon Deog-Bok , Ha Tae-Yong , Cho Sung-Hun , Oh Ki-Bong
Published: January 2004

The left-sided gallbladder is very rare, but it is often accompanied by multiple anomalies of the liver, by which living donor hepatectomy cannot be feasible or becomes difficult. We have experienced 3 donors with a left-sided gallbladder out of 642 living donors. The first case was a male donor showing bifurcating portal anomaly with intrahepatic right portal vein confluence and extremely low bifurcation of the bile ducts. The right lobe was retrieved and implanted to his father. The second case was a male donor revealing trifurcating portal anomaly with separate right posterior portal branch and replacing right posterior hepatic artery. The right posterior segment graft was retrieved and implanted to his uncle. The third case was a male volunteer in whom the anterior portion of the medial segment was fed by an aberrant branch of the right anterior segment glisson. The small left lobe was retrieved and implanted simultaneously with another living donor’s left lobe graft in the form of a dual living donor liver transplantation. There was no donor morbidity or recipient complication. Although there is a high possibility of diverse liver anomalies in living donors with a left-sided gallbladder, complete preoperative evaluation and mapping of the multiple anatomical variations may make certain types of living donor hepatectomy feasible. (Liver Transpl 2004;10:141-146.)
Pseudohypocalcemia after magnetic resonance imaging with gadolinium in patients with cirrhosis
Authors: Kefalas Costas H. , Murray Natalie G.B. , Aguanno James J. , Dockery William D. , Weinstein Jeffrey S. , Anderson Katherine , Klintmalm Goran B.
Published: January 2004

Hypocalcemia in patients with cirrhosis may be due to a number of causes. We noted a relationship between injection with gadodiamide, a particular gadolinium chelate, during magnetic resonance imaging of the liver and the development of a falsely low serum total calcium level in a patient with cirrhosis. A cross-reference and retrospective chart review identified 10 additional patients in whom this phenomenon was noted. We describe the temporal relationship and clinical characteristics of these patients. Pseudohypocalcemia following magnetic resonance imaging with gadodiamide contrast should be considered in the differential diagnosis of hypocalcemia in patients with cirrhosis. (Liver Transpl 2004;10:136-140.)
Incomplete improvement of visuo-motor deficits in patients with minimal hepatic encephalopathy after liver transplantation
Authors: Mechtcheriakov Sergei , Graziadei Ivo W. , Mattedi Michael , Bodner Thomas , Kugener André , Hinterhuber Hartmann H. , Marksteiner Josef , Vogel Wolfgang
Published: January 2004

Previous studies have suggested reversibility of minimal hepatic encephalopathy in patients with liver cirrhosis after liver transplantation (LT), however, this topic is controversially discussed. We investigated this issue in a prospective study on liver cirrhotic patients listed for LT. Patients were investigated before and after liver transplantation (on average 21 months later) using a neuropsychological test battery which measured visuo-constructive and visuo-motor ability, verbal fluency, and memory function. To assess visuo-motor and visuo-constructive functions, we performed 4 tests: Rey Complex Figure Test copy, trail making tests A and B, and digital symbol test. The average percentile score of the tests, arbitrarily named the visuo-motor and visuo-constructive performance score (VMCP), was calculated. After LT, the patients did not demonstrate a significant increase of VMCP (P = .29) and additionally showed significantly lower VMCP score (P = .041) compared to control group. Analysis of individual responses showed that only 7 of 14 patients improved their VMCP values after LT. These data indicate that the cirrhosis-associated visuo-motor deficits subside or disappear only in some of the patients after LT, whereas a significant number of patients show no improvement of the visuo-motor and visuo-constructive function. We concluded that monitoring of cognitive and visuo-motor functioning is important for the post-transplant rehabilitation of patients with liver cirrhosis. (Liver Transpl 2004;10:77-83.)
Hepatic arterial anatomy for right liver procurement from living donors
Authors: Kishi Yoji , Sugawara Yasuhiko , Kaneko Junichi , Akamatsu Nobuhisa , Imamura Hiroshi , Asato Hirotaka , Kokudo Norihiro , Makuuchi Masatoshi
Published: January 2004

Living donor liver transplantation (LDLT) using right liver grafts is now widely performed. Anatomic classifications of the hepatic artery for right liver procurement, however, are limited. In this study, celiac and mesenteric angiograms of 223 consecutive living donors in a single institution were evaluated. Details of the arterial anastomosis and results were reviewed in 72 patients who underwent primary LDLT using right liver grafts. There was a 6% incidence of hepatic arterial bifurcations that might provide multiple orifices in a right liver graft. Only one right liver graft (1%) had multiple arterial orifices. Single arterial anastomosis without interposition was possible in all patients with right liver grafts and none of them were complicated with hepatic arterial thrombosis. Single arterial anastomosis, therefore, has a high probability of success in right liver graft implantation. (Liver Transpl 2004;10:129-133.)
Quantitative assessment of fibrinogen cross-linking by εaminocaproic acid in patients with end-stage liver disease
Authors: Quach Thien , Tippens Melissa , Szlam Fania , Van Dyke Rebecca , Levy Jerrold H. , Csete Marie
Published: January 2004

Analysis of the effectiveness of antifibrinolytic therapy for liver transplant recipients is hampered by lack of quantitative assays for assessing drug effects. We adapted chemical engineering tools used in polymerization studies to quantify fibrinogen cross-linking by plasma from liver transplant patients obtained before and after εaminocaproic acid (EACA) therapy. A target fluorescein isothiocyanate-fibrinogen (FITC—fibrinogen) molecule was constructed; it fluoresces in a quantifiable pattern when in solution, and undergoes cross-linking in the presence of plasmin inhibitors. Cross-linking quenches the fluorescent signal, and the quenching is a quantifiable endpoint. Thus fluorescence from this reporter molecule can be used to assess functional improvement in fibrinogen cross-linking as a result of antifibrinolytic therapies, and it is sensitive to picomolar amounts of plasmin inhibitors and activators. Cross-linking of FITC-fibrinogen by patient plasma, before and after EACA therapy, was assessed using fluorescence spectrometry. Fluorescence patterns from FITC-fibrinogen indicated no significant cross-linking of the target fibrinogen as a consequence of EACA in posttreatment plasma. When the fibrinogen-FITC target was assayed without plasma in the presence of EACA at concentrations that bracket therapeutic levels (100 and 400 μg/ml), significant fluorescence quenching (target FITC-fibrinogen cross-linking) was achieved. These results suggest that fibrinogen-FITC fluorescence is sensitive enough to detect EACA activity in clinically relevant ranges, but that EACA given in usual doses is insufficient to promote fibrinogen cross-linking in patients with end-stage liver disease. (Liver Transpl 2004;10:123-128.)
Liver transplantation for hepatocellular carcinoma: The MELD impact
Authors: Sharma Pratima , Balan Vijayan , Hernandez Jose L. , Harper Ann M. , Edwards Erick B. , Rodriguez-Luna Hector , Byrne Thomas , Vargas Hugo E. , Mulligan David , Rakela Jorge , Wiesner Russell H.
Published: January 2004

The new allocation policy of the United Network of Organ Sharing (UNOS) based on the model for end-stage liver disease (MELD) gives candidates with stage T1 or stage T2 hepatocellular carcinoma (HCC) a priority MELD score beyond their degree of hepatic decompensation. The aim of this study was to determine the impact of the new allocation policy on HCC candidates before and after the institution of MELD. The UNOS database was reviewed for all HCC candidates listed between July 1999 and July 2002. The candidates were grouped by two time periods, based on the date of implementation of new allocation policy of February 27, 2002. Pre-MELD candidates were listed for deceased donor liver transplantation (DDLT) before February 27,2002, and post-MELD candidates were listed after February 27, 2002. Candidates were compared by incidence of DDLT, time to DDLT, and dropout rate from the waiting list because of clinical deterioration or death, and survival while waiting and after DDLT. Incidence rates calculated for pre-MELD and post-MELD periods were expressed in person years. During the study, 2,074 HCC candidates were listed for DDLT in the UNOS database. The DDLT incidence rate was 0.439 transplant/person years pre-MELD and 1.454 transplant/person years post-MELD (P < 0.001). The time to DDLT was 2.28 years pre-MELD and 0.69 years post-MELD (P < 0.001). The 5-month dropout rate was 16.5% pre-MELD and 8.5% post-MELD (P < 0.001). The 5-month waiting-list survival was 90.3% pre-MELD and 95.7% post-MELD (P < 0.001). The 5-month survival after DDLT was similar for both time periods. The new allocation policy has led to an increased incidence rate of DDLT in HCC candidates. Furthermore, the 5-month dropout rate has decreased significantly. In addition, 5-month survival while waiting has increased in the post-MELD period. Thus, the new MELD-based allocation policy has benefited HCC candidates. (Liver Transpl 2004;10:36-41.)
Liver transplantation and health-related quality of life: Scoring differences between men and women
Authors: Cowling Terianne , Jennings Linda W. , Goldstein Robert M. , Sanchez Edmund Q. , Chinnakotla Srinath , Klintmalm Goran B. , Levy Marlon F.
Published: January 2004

Orthotopic liver transplantation (OLT) is the treatment of choice for end-stage liver disease of various etiologies. Its use, however, remains limited due to the scarcity of donor organs. Measures to assess health-related quality of life (HRQOL) are increasingly being implemented to examine the efficacy of medical therapies involving scarce resources. HRQOL was assessed and compared between 88 male and 61 female patients before and after liver transplantation. Data were gathered from subjects having completed a questionnaire pre-OLT, and again at 1 year and 2 years post-OLT. This questionnaire, developed specifically for OLT patients, contains at its core questions derived from several well-established instruments measuring health status and HRQOL. Male OLT recipients reported a higher degree of overall HRQOL than that reported by female OLT recipients, both before and after OLT. When controlling for disparity in education between the sexes, findings revealed that among the lesser educated (≤12 years), men and women scored similarly, while among the more educated (>12 years), men scored higher than women. Employment findings revealed a higher percentage of men working before transplant and at 1-year post-OLT when compared with women. At 2 years post-OLT, men and women exhibited similar employment rates. Male OLT recipients report a higher level of overall HRQOL than that reported by female OLT recipients, both before and after liver transplantation. Education appears to significantly affect HRQOL and may account for, at least in part, differences in reported HRQOL between male and female OLT recipients. (Liver Transpl 2004;10:88-96.)