Electroporation-mediated interleukin-12 gene therapy for hepatocellular carcinoma in the mice model.

Applications of nonviral vectors for gene transfer into tumors in vivo have been limited by the relatively low expression levels of the transferred gene. The aim of this study is to evaluate the efficacy of electroporation-mediated interleukin-12 (IL-12) gene therapy for hepatocellular carcinoma (HCC). First, we investigated the optimal conditions of electric pulses (voltage, pulsing duration, numbers of shocks) of in vivo electroporation for gene transfer into HCC established by s.c. implantation of MH134 cells to C3H mice. This process made use of plasmid DNA that express the luciferase gene. We concluded that the optimal conditions for the electric pulses are as follows: voltage at 150 V; pulsing duration at 50 ms; nonpulsing duration at 950 ms; and the number of shocks at 10. Second, we tried to treat s.c. HCC by electroporation using plasmid DNA that expresses the murine interleukin-12 (mlL-12) gene. Intratumoral administration of the mIL-12 vector elevated serum IL-12 and IFN-gamma and significantly inhibited the growth not only of HCC into which the mIL-12 vector had been directly transferred, but also of the distant HCC. In addition, intratumoral administration of the mIL-12 vector inhibited spontaneous lung metastasis and delayed establishment of HCC injected 3 days after mIL-12 gene therapy. The IL-12 gene therapy induced more lymphocyte infiltration by NK cells, CD3+ cells, and Mac-1 positive cells into the tumor and reduced the number of microvessels. Therefore, more terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling-positive tumor cells were found. These results demonstrate that gene therapy for HCC by electroporation in vivo using IL-12 is very efficient and is thus promising for further clinical trial.

Cyclophosphamide-induced tolerance in rat orthotopic liver transplantation.

BACKGROUND: We previously established a cyclophosphamide (CP)-induced tolerance system in rodent skin graft models. In this study, we applied this system to rat liver transplantation. METHODS: Lewis recipients were inoculated on day -2 with spleen and bone marrow cells (SC+BMC) from Dark Agouti (DA) donors, followed by 100 mg/kg CP on day 0. On day 25, DA livers were orthotopically grafted. We assessed the alloresponses to the donors of the long-term surviving recipients, using the second skin grafting and in vitro assay. RESULTS: The recipients that had been treated with SC+BMC and CP survived for more than 165 days. None of control group that received SC+BMC alone (mean survival times [MST]=13.8 days), CP alone (MST=40.0), SC+BMC from third-party PVG rats and CP (MST=45.0), or no treatment (MST=13.8) survived over 50 days. The donor-specific tolerance was confirmed by second skin grafts onto recipients with permanent DA liver grafts, which accepted DA skins (MST>75) but not PVG (MST=8.3). However, the lymphocytes from the tolerant recipients showed alloresponse to DA in vitro. To investigate whether the T helper type 2 deviation contributed to this "split tolerance," we assessed the production of cytokines in mixed lymphocyte reaction. Interleukin 2 and interferon-gamma were detected but interleukin 4 was not. CONCLUSIONS: These data showed that this protocol induced split tolerance in rat liver transplantation and, furthermore, the mechanism of split tolerance was not due to T helper 2 deviation.

Beneficial effects of 3-dimensional visualization on hepatic vein reconstruction in living donor liver transplantation using right lobe graft.

BACKGROUND: Recently, virtual operation planning and navigation systems have been introduced in the field of neurosurgery and orthopedic surgery. We report here the beneficial effects of 3-dimensional (3D) visualization on hepatic venous reconstruction in living donor liver transplantation (LDLT) using right lobe graft. METHODS: 3D-image reconstruction of the liver was rendered with 3-mm slices of helical computed tomography (CT) data using zioM900 (Zio Software Inc., Tokyo, Japan). To understand the anatomy of the donor's vessels and design an operation plan, a picture of the vessels in and around the liver was reconstructed. RESULTS: The 3D image demonstrated two short hepatic veins next to the inferior right hepatic vein (IRHV) as well as a large IRHV. The 3D image showed a more precise diameter of the right hepatic vein (RHV) and the IRHV and a more accurate distance between the two hepatic veins than did images measured by 2-dimensional CT. This preoperative information allowed the donor surgeon to dissect the inferior vena cava (IVC) and hepatic veins with reduced blood loss because of reduced risk of injury to the blood vessels. The 3D image revealed that both the RHV and the IRHV branched off at the same angle from the cylindrical IVC. Preoperative planning based on this information secured smooth anastomosis. CONCLUSIONS: 3D visualization is useful for hepatic venous reconstruction of the recipient as well as for donor surgery in LDLT using right lobe graft.

Running mileage, movement mileage, and fitness in male U.S. Navy recruits.

PURPOSE: The purpose of this study was to determine whether there is a relationship between overall fitness improvement and varying amounts of running and movement mileage. METHODS: Subjects were male U.S. Navy recruits (N = 1703, 25 divisions), ages 17-35 yr (mean age = 20.1 +/- 2.9 yr), who attended boot camp from April 1996 through August 1996. During the first week of training, recruits performed a 1.5-mile run to determine baseline fitness levels. The results from the initial run were compared with a final 1.5-mile run conducted 6 wk later. RESULTS: Based on an age-adjusted fitness scale for a 1.5-mile run time, about one third of the recruits began recruit training in "Excellent-Superior" condition (N = 558), one third began in "Good" condition (N = 582), and one third began in "Poor-Fair" condition (N = 563). Running mileage among divisions ranged from 11.5 to 43.5 miles for the entire 7-wk training period (mean = 22.7 +/- 7.2 miles; 8-22 run days, mean = 13 +/- 4 d). In addition to running, the divisions accumulated many movement miles (110-202 miles; mean = 145 +/- 26 miles) while marching in formation. Recruits who began training in Poor-Fair condition improved the most with an average decrease in run time of 1:55 +/- 1:06 min (15.6% improvement). The Good group improved by 47 +/- 37 s (7.3% improvement), and the Excellent-Superior group improved by 17 +/- 32 s (2.9% improvement). CONCLUSION: The magnitude of fitness improvement, as measured by run time improvement, was directly related to baseline fitness level but not related to movement mileage or high-intensity run mileage accrued during training.

Living-related auxiliary partial orthotopic liver transplantation for primary sclerosing chonangitis--subsequent removal of the native liver.

In Japan, living-related auxiliary partial orthotopic liver transplantation (APOLT) is mainly indicated for small-for-size grafts. We present the case of a 24 year-old patient with primary sclerosing cholangitis (PSC) who underwent a living-related auxiliary partial orthotopic liver transplantation for a small-for-size graft, that was subsequently excised. During the transplantation procedure, the native liver was freed from the surrounding tissues, and was only connected to the body by the right hepatic artery and right hepatic vein. The auxiliary extended left lobe graft, corresponding to 22% of the estimated recipient liver volume, was orthotopically transplanted after extended left lobectomy of the recipient native liver. Post-transplant CT-volumetry showed rapid increase of the graft volume with atrophy of the native liver, and GSA scintigraphy showed dominant function of the graft. Although hyper-bilirubinemia was prolonged by the removal of the native liver on the 18th post-transplantation day, it gradually subsided after plasmapheresis was performed twice, and the patient was discharged on the 77th post-transplantation day. We conclude, based on this case, that subsequent removal of the native liver is necessary in APOLT for patients with potential hepatic malignancies. The optimal timing of the removal of the remnant native liver should be determined based on CT-volumetry, GSA scintigraphy, and the liver biopsy specimen of the graft.

Intra-operative hyaluronic acid clearance predicts postoperative graft function in living related liver transplantation.

Although intra-operative hyaluronic acid (HA) clearance has been reported to be a predictive parameter for early graft function after the implantation of a cadaveric hepatic graft, it has yet to be evaluated as a parameter in assessing graft function in living related liver transplantation. The aim of this study was to evaluate whether intra-operative HA clearance can be a predictive parameter of early graft function in living related liver transplant patients. Eight consecutive patients, who underwent a living related liver transplantation, were entered into the study. The HA clearance 180 min after reperfusion of the graft was evaluated. Significantly higher serum HA levels were found in the patients with fulminant hepatitis than in the patients with non-fulminant hepatitis before operation (P < 0.01), just before reperfusion (P < 0.01), and 180 min after reperfusion (P < 0.05). The HA clearance correlated with the peak total bilirubin within 5 postoperative days (P < 0.05) and the lactic acid one day after operation (P < 0.01). The intra-operative HA clearance serves as a sensitive parameter for assessing the postoperative graft function after the implantation of the new liver. Based on our findings, measuring the HA clearance was thus found to be clinically useful in the assessment of graft function in living related liver transplantation.

One-year follow-up evaluation of the sexually transmitted diseases/human immunodeficiency virus intervention program in a marine corps sample.

Although a substantial number of studies have been conducted to evaluate the impact of various human immunodeficiency virus (HIV) prevention programs, most of them have focused on civilian populations. There is a clear need to develop and evaluate sexually transmitted diseases (STD)/HIV prevention programs designed specifically for U.S. military populations. The objective of the present study was to determine whether a behavioral intervention known as the STD/HIV Intervention Program (SHIP) would have a sustained positive impact on the behavior of a sample of Marines. A 1-year follow-up telephone interview was administered to (1) Marines who participated in the SHIP course (intervention group), and (2) a quasi-control group of Marines who were not exposed to the SHIP course. The intervention and control groups differed significantly in the percentage of the time they had used condoms during the past year. The intervention participants reported using condoms a greater percentage of the time than the nonparticipants.

[Low dose cyclosporin treatment for ocular lesions of Behçet's disease].

Cyclosporin is an immunosuppressant that has recently been used for the treatment of Behçet's disease. We evaluated the clinical effect of low doses of cyclosporin (5 mg/kg/day) on 20 patients of Behçet's disease with refractory ocular involvement. The therapeutic effect of cyclosporin was judged by the frequency of ocular inflammatory attacks. It was effective in more than 70% of the patients. On the other hand, it had some adverse side effects such as renal dysfunction, indigestion, and neurological disorders. Most side effects were mild with low dose treatment, but neurological side effects were similar in frequency and severity to those seen with 10 mg/kg/day usage. These findings indicate that low dose cyclosporin (5 mg/kg/day) administration may be acceptable as a standard plan for treatment of Behçet's disease, but that we have to use cyclosporin with great attention to its side effects, especially neurological symptoms.

[Living donor liver transplantation in Kyushu University].

We performed living donor liver transplantation (LDLT) for 40 patients at Kyushu University Hospital, Fukuoka Japan during the period from October 1996 to April 2000. The patients consisted of 32 adults and 8 children with a mean age of 35.8 years (range: 1 year and 10 months to 65 years old). The underlying liver diseases of the 40 patients included the fulminant hepatic failure (n = 14), biliary atresia (n = 7), liver cirrhosis (HCV) (n = 6), primary biliary cirrhosis (n = 5), primary sclerosing cholangitis (n = 2), familiar amyloidotic polyneuropathy (n = 2), Alagille syndrome (n = 1), glycogen storage disease (n = 1), huge hepatic hemangiomas (n = 1), and Wilson's disease (n = 1). All liver grafts were obtained from each patient's family members except for one domino transplant donor's case, comprised of 13 parents, 13 sons and daughters, 11 brothers and sisters, and 3 wives. The donors are presently all doing well. The patient survival rate is presently 92.5%.

Central and peripheral noradrenergic tone in primary hypertension.

The contents of norepinephrine (NE), epinephrine (E), dopamine (DA), normetanephrine (NMN), and 4-hydroxy-3-methoxyphenylethylene glycol (MHPG) were measured in the plasma and cerebrospinal fluid (CSF) of 66 patients with primary hypertension and 24 patients with normal blood pressure and minor neurological disorders. Plasma and CSF NE and NMN concentrations were raised in the hypertensive patients. The plasma and CSF NE levels and arterial blood pressure of a small subset of hypertensive patients were normalized after clonidine therapy. In hypertensive patients the content of DA was lower and the ratio of NE/DA was greater; CSF and plasma NE contents were related to the level of arterial blood pressure; and the content of MHPG in CSF was linked strongly with NE content in plasma and CSF and to the level of arterial blood pressure. Thus both central sympathetic nerve tone and peripheral sympathetic nerve tone were enhanced in young patients with uncomplicated hypertension. The elevated levels of neurohormones and their metabolites in some patients with primary hypertension may be related to increased synthesis and release of neural NE and may be pathogenic in the blood pressure elevation.

A measure of the intensity of response to alcohol in a military population.

Heavy drinking and associated problems are relatively common in young men, including those in a military setting. This article explores characteristics of alcohol intake and associated difficulties and their relationship to a self-report of the usual intensity of response to alcohol in a sample of U.S. Marines. Two questionnaires related to demography and alcohol use histories, along with a simple, 12-item self-report measure of the usual number of drinks to experience an effect (the Self-Rating of the Effects of Alcohol, SRE) were administered to 1320 U.S. Marines. The sample had an average age of 22 years, 78% were Caucasian, and 92% were enlisted personnel. The relationships and correlations among drinking characteristics and problems and the usual number of drinks for an effect were determined. These subjects drank an average of 6 days per month, consuming an average of almost six drinks per drinking day, and reported more than three times per month in which they consumed six or more drinks per occasion. Consistent with studies of other populations, the SRE measures of intensity of response to alcohol showed a positive correlation with both drinking practices and problems, with the latter remaining significant even after controlling for recent drinking practices. The prodigious level of alcohol intake and associated problems, along with the SRE scores, indicate that the Marine Corps personnel are at especially high risk for alcohol-related life problems. These data also support the potential usefulness of the SRE both in identifying individuals likely to have more severe alcohol profiles and in educating individuals regarding their levels of risk for alcohol abuse and dependence.