Akiyoshi Kanazawa

Inhibition of tumor necrosis factor-induced apoptosis in transgenic mouse liver expressing creatine kinase

Abstract: Background: The mitochondrion acts as a pivotal decision center in many types of apoptotic responses. To clarify the effects of the enhanced mitochondrial function on tumor necrosis factorα (TNFα)‐induced apoptosis, we studied hepatic injuries in transgenic mice whose livers express creatine kinase (CK).

Open versus laparoscopic resection of primary tumor for incurable stage IV colorectal cancer: a large multicenter consecutive patients cohort study.

Objective:To investigate the hypothesis that laparoscopic primary tumor resection is safe and effective when compared with the open approach for colorectal cancer patients with incurable metastases. Background:There are only a few reports with small numbers of patients on laparoscopic tumor resection for stage IV colorectal cancer. Methods:Data from consecutive patients who underwent palliative primary tumor resection for stage IV colorectal cancer between January 2006 and December 2007 were collected retrospectively from 41 institutions. Short- and long-term outcomes were compared between patients who underwent laparoscopic or open resection. Results:A total of 904 patients (laparoscopic group: 226, open group: 678) with a median age of 64 years (range: 22–95) were included in the analysis. Conversion was required in 28 patients (12.4%) and the most common reasons for conversion (23/28: 82%) were bulky or invasive tumors. There was no 30-day postoperative mortality in either group. The complication rate (NCI-CTCAE grade 2–4) after laparoscopic surgery (17%) was significantly lower than that after open surgery (24%) (P = 0.02), and the difference was greater (4% vs 12%; P < 0.001) when we limited the analysis to severe (≥grade 3) complications. The median length of postoperative hospital stay in the laparoscopic group was significantly shorter than that in the open group (14 vs 17 days; P = 0.002). In univariate analysis, overall survival for the laparoscopic group was significantly better than that for open surgery (median survival time: 25.9 vs 22.3 months, P = 0.04), although no difference was apparent in multivariate analysis. Conclusions:Compared with open surgery, laparoscopic primary tumor resection has advantages in the short term and no disadvantages in the long term. It is a reasonable treatment option for certain stage IV colorectal cancer patients with incurable disease.

Humoral injury in porcine livers perfused with human whole blood

BACKGROUND We investigated the influence of humoral injury during xenoperfusion of porcine livers by human blood. METHODS The porcine livers were perfused under physiological conditions for 9 hr. The perfusates consisted of porcine whole blood in group 1, human whole blood in group 2, and human whole blood with soluble complement receptor type 1 (300 microg/ml) in group 3. RESULTS Liver enzyme release and serum hemoglobin in group 2 increased significantly after 3 hr of xenoperfusion, compared with those in group 1 and group 3 (P<0.05). Severe histological damage with minimal cellular infiltration was observed in group 2 after 6 hr of xenoperfusion, but was present only at trace levels in group 1 and group 3. In group 2, von Willebrand factor, a possible target of natural antibodies, was induced on sinusoidal endothelial cells after 3 hr of xenoperfusion, correlating with diffuse deposition of human IgM and membrane attack complex. In group 3, von Willebrand factor, human IgM, and membrane attack complex staining in the intralobular region were present at trace levels. In group 3, the indocyanine green removal capacity, representing hepatocyte function, was significantly higher than in group 2 (P<0.05). CONCLUSIONS Based on these results, we suggest that humoral injury is a major cause of liver damage during liver xenoperfusion. The pattern of humoral injury in xenoperfused livers may be attributed to anatomical features of the liver and unique responses of sinusoidal endothelial cells to xenoperfusion.

Evaluation of ammonia and lidocaine clearance, and galactose elimination capacity of xenoperfused pig livers using a pharmacokinetic analysis.

BACKGROUND We introduced the pharmacokinetic method into the functional evaluation of xenogeneic extracorporeal liver perfusion as an artificial liver assist device, and examined the influence of xenogeneic humoral injury on the metabolic function of xenoperfused pig livers. METHODS Isolated pig livers were perfused with fresh porcine blood (group 1; n=5) or fresh human blood (group 2; n=5) for 9 hr. Clearance (CL) of ammonia and lidocaine, and galactose elimination capacity (Vmax) were determined at three points during the perfusion using a one-compartment pharmacokinetic model. RESULTS Concentrations of ammonia and lidocaine decreased exponentially and those of galactose decreased linearly after a bolus injection in both groups. A one-compartment model provided satisfactory curve fittings for these test substances. No decreases of ammonia CL, lidocaine CL, or galactose Vmax were observed until 9 hr in either group. No differences were observed between the two groups with respect to these metabolic functions. In group 1, only slight interlobular edema was observed at 9 hr. In group 2, membrane attack complex was diffusely deposited at 3 hr and severe interlobular damage was histologically observed at 9 hr, although hepatocellular damage was minimal even at 9 hr. Alpha glutathione S-transferase and mitochondrial aspartate aminotransferase were comparable between the two groups. CONCLUSIONS Pharmacokinetic analysis allowed the evaluation of ammonia CL, lidocaine CL, and galactose Vmax of the perfused pig livers. Despite xenogeneic humoral injuries, the xenoperfused livers maintained these metabolic functions at the same levels as the alloperfused livers for 9 hr.

Extension of the indication for living related liver transplantation from children to adults based on resolution of graft size mismatch in relation to tissue oxygenation and metabolic load: a case report

Abstract  We extended the indication for living related partial liver transplantation from pediatric to adult cases. Our first case was a 49‐year‐old woman with primary biliary cirrhosis. Her sisters left lobe, weighing 280 g, was employed as a graft, and the graft weight/recipients body weight ratio was calculated as 0.59 %. To decrease the metabolic load to the relatively small graft, the total bilirubin was decreased from a maximum value of 75.0 mg/dl to the most recent preop‐erative value of 36.2 mg/dl by plasma exchange. Intraoperative recovery of tissue oxygenation and its heterogeneity were satisfactory due to a relatively high blood supply. A postoperative decrease in bilirubin and increase in cholesterol esterifi‐cation were facilitated, concomitant with regeneration of the graft, which weighed 280 g, to 860 c at 3 weeks. Linear regression analysis with respect to tissue oxygenation and metabolic capacity obtained in pediatric cases were applied to this adult case.

Multicenter safety study of mFOLFOX6 for unresectable advanced/recurrent colorectal cancer in elderly patients

BackgroundCombination chemotherapy with oxaliplatin plus 5-fluorouracil/leucovorin (FOLFOX) has become a standard regimen for colorectal cancer. An increase of adverse events with combination chemotherapy is predicted in elderly patients, and it remains controversial whether they should receive the same chemotherapy as younger patients. Accordingly, this study of modified FOLFOX6 (mFOLFOX6) therapy was performed to compare its safety between elderly and non-elderly patients.MethodsWe prospectively studies 14 non-elderly patients aged <70 years old and 8 elderly patients aged ≥ 70 years with unresectable advanced/recurrent colorectal cancer who received mFOLFOX6 therapy during the period from March 2006 to March 2007. Adverse events and the response to treatment were compared between the elderly and non-elderly groups.ResultsThe main adverse events were neutropenia and peripheral neuropathy, which occurred in 62.5% (≥ grade 3) and 87.5% (≥ grade 1) of elderly patients. The grade and frequency of adverse events were similar in the elderly and non-elderly groups. In some patients with neutropenia, treatment could be continued without reducing the dose of oxaliplatin by deleting bolus 5-fluorouracil. A correlation was found between the cumulative dose of oxaliplatin and the severity of neuropathy, and there were 2 elderly and 3 younger patients in whom discontinuation of treatment was necessary due to peripheral neuropathy. The median number of treatment cycles was 10.0 and 9.5 in the non-elderly and elderly groups, respectively. The response rate was 60.0% in the non-elderly and 50.0% in the elderly group, while the disease control rate was 100% and 83.3% respectively, showing no age-related difference.ConclusionmFOLFOX6 therapy was well-tolerated and effective in both non-elderly and elderly patients. However, discontinuation of treatment was sometimes necessary due to peripheral neuropathy, which is dose-limiting toxicity of this therapy.

The protective role of Kupffer cells in humoral injury of xenoperfused rat livers

BACKGROUND The role of Kupffer cells in a hepatic xenograft rejection is still unclear. We investigated the effect of blocking Kupffer cells on xenogeneic humoral injury using rat livers as the xenoperfusion models. METHODS Rat livers were perfused with fresh human blood after pretreatment either with normal saline (group 1; n = 8) or with gadolinium chloride (GdCl3) solution (group 2; n = 8). Tissue injury was evaluated by alanine aminotransferase release and histological examination. Tumor necrosis factor-alpha (TNF-alpha) production from rat livers was measured by enzyme-linked immunosorbent assay and also examined by immunohistochemistry. In addition, Kupffer cells were isolated after pretreatment either with normal saline or with GdCl3 solution and incubated with human serum. Localization of human C3 and IgM was examined by immunofluorescence. RESULTS Alanine aminotransferase release in group 2 was significantly higher than in group 1 (P = 0.015). Histological examination revealed more severe tissue injury in group 2. The mean TNF-alpha level was not significantly different between the two groups. In immunohistochemistry, TNF-alpha was positive primarily on vascular endothelial cells in both groups. Immunofluorescence of saline-treated Kupffer cells showed an uptake of human C3 in the cytoplasm, whereas no uptake was observed in GdCl3-treated cells. The uptake of human IgM did not differ between the two groups. CONCLUSIONS These results suggest that Kupffer cells have a protective role in preventing xenogeneic humoral injury. Their ability to absorb xenogeneic complements may contribute to this protective mechanism.

Conversion to complete resection with mFOLFOX6 with bevacizumab or cetuximab based on K‐ras status for unresectable colorectal liver metastasis (BECK study)

Patients with colorectal liver metastasis (CRLM) might be down‐staged by chemotherapy from an initially unresectable stage to a resectable stage. Because the tumor response to preoperative chemotherapy has been correlated with resection rate, the improved efficacy from the concept that only the patients without K‐ras mutations receive an anti‐EGFR antibody might be expected to increase the conversion rate. The purpose of this study is to evaluate the conversion rate from unresectable CRLM to complete resection.

Study protocol of the B-CAST study: a multicenter, prospective cohort study investigating the tumor biomarkers in adjuvant chemotherapy for stage III colon cancer

BackgroundAdjuvant chemotherapy for stage III colon cancer is internationally accepted as standard treatment with established efficacy. Several oral fluorouracil (5-FU) derivatives with different properties are available in Japan, but which drug is the most appropriate for each patient has not been established. Although efficacy prediction of 5-FU derivatives using expression of 5-FU activation/metabolism enzymes in tumors has been studied, it has not been clinically applied.Methods/designThe B-CAST study is a multicenter, prospective cohort study aimed to identify the patients who benefit from adjuvant chemotherapy with each 5-FU regimen, through evaluating the relationship between tumor biomarker expression and treatment outcome. The frozen tumor specimens of patients with stage III colon cancer who receives postoperative adjuvant chemotherapy are examined. Protein expression of thymidine phosphorylase (TP), dihydropyrimidine dehydrogenase (DPD), epidermal growth factor receptor (EGFR), and vascular endothelial growth factor (VEGF) are evaluated using enzyme-linked immunosorbent assay (ELISA). mRNA expression of TP, DPD, thymidylate synthase (TS) and orotate phosphoribosyl transferase (OPRT) are evaluated using reverse transcription polymerase chain reaction (RT-PCR). The patients’ clinical data reviewed are as follow: demographic and pathological characteristics, regimen, drug doses and treatment duration of adjuvant therapy, types and severity of adverse events, disease free survival, relapse free survival and overall survival. Then, relationships among the protein/mRNA expression, clinicopathological characteristics and the treatment outcomes are analyzed for each 5-FU derivative.DiscussionA total of 2,128 patients from the 217 institutions were enrolled between April 2009 and March 2012. The B-CAST study demonstrated that large-scale, multicenter translational research using frozen samples was feasible when the sample shipment and Web-based data collection were well organized. The results of the study will identify the predictors of benefit from each 5-FU derivative, and will contribute to establish the “personalized therapy” in adjuvant chemotherapy for colon cancer.Trial registrationClinicalTrials.gov: NCT00918827, UMIN Clinical Trials Registry (UMIN-CTR) UMIN000002013

Short- and long-term outcomes following laparoscopic palliative resection for patients with incurable, asymptomatic stage IV colorectal cancer: A multicenter study in Japan

Objective: This retrospective multicenter study compared short- and long-term results between Japanese patients with asymptomatic stage IV colorectal cancer who underwent palliative laparoscopic surgery (LS) versus those who underwent conventional open surgery (OS). Methods: Among 968 patients treated for stage IV colorectal cancer from January 2006 to December 2007 in 41 surgical units that were participating in the Japan Society of Laparoscopic Colorectal Surgery group, we studied 398 patients who received palliative resection of their asymptomatic primary colorectal tumor. Results: We analyzed data from patients undergoing LS (LS group, n=106) and OS (OS group, n=292). Fourteen (13.2%) LS group patients were converted to OS. Although the differences between groups for postoperative complications were not significant, the mean time to solid food intake and postoperative length of hospital stay for the LS group were significantly shorter than those for the OS group (2 vs. 3 days, p<0.0001; 13 vs. 16 days, p<0.0001, respectively). The LS group patients experienced a longer median survival time than that of the OS group (24.5 vs. 23.9 months, p=0.0357). Conclusions: Laparoscopic palliative resection (LS) offers advantages for short-term outcomes and no disadvantages for long-term outcomes. The use of laparoscopic procedures to treat asymptomatic, incurable stage IV colorectal cancer appears to be acceptable.