Zinc transporter 1 expression in hepatocellular carcinoma correlates with prognosis: A single-center retrospective study.

OBJECTIVES: Zinc is crucial in the pathogenesis of hepatocellular carcinoma; however, no reports have examined its association with clinical parameters and zinc transporter 1 (ZNT1) expression intensity. This study aimed to assess the association between ZNT1 expression and prognosis in patients with hepatocellular carcinoma. METHODS: This retrospective study included 65 patients who underwent surgical hepatocellular carcinoma resection at a single center between January 2011 and June 2015. ZNT1 expression on hepatocellular carcinoma cells from specimens was assessed using immunohistochemistry, and the relationship between its intensity and various clinical indexes was examined with univariate and multivariable analyses and the Mann-Whitney U, Kruskal-Wallis, Bonferroni, and log-rank tests. RESULTS: ZNT1 expression on the hepatocellular carcinoma cell membrane was negative in 31 patients and positive in 34 patients, including nine patients showing strongly positive expression. Patients with and without ZNT1 expression had similar blood zinc concentrations, α-fetoprotein levels, protein induced by vitamin K absence-antagonist-II levels, gross classification, maximal tumor diameters, and background liver disease. The blood zinc concentrations were significantly lower in patients with strongly positive ZNT1 expression (57.0 ± 22.1 µg/dL) than in those with positive ZNT1 expression (71.1 ± 14.2 µg/dL; P = 0.015) or those with no ZNT1 expression (72.9 ± 14.1 µg/dL; P = 0.043). Overall survival was significantly shorter in ZNT1-expressing patients than in non-expressing patients (log-rank test, P = 0.024). Multivariable analysis using the Cox proportional hazards model identified maximal tumor diameter (hazard ratio, 1.018; 95% confidence interval, 1.002-1.034; P = 0.026) and ZNT1 expression status (hazard ratio, 2.082; 95% confidence interval, 1.196-3.621; P = 0.010) as prognostic contributing factors.

Ultrasound-guided infraclavicular axillary vein puncture is effective to avoid pinch-off syndrome: a long-term follow-up study.

PURPOSE: Pinch-off syndrome (POS) is a serious complication encountered during the long-term management of totally implantable access ports (TIAPs). The aim of this study was to examine the effect of ultrasound-guided infraclavicular axillary vein puncture to avoid POS in patients with long-term use of a TIAP. METHODS: This was a retrospective review of 207 consecutive TIAPs: one hundred devices implanted using an anatomical landmark technique were used as historical controls (Landmark group), while 107 devices were implanted using an ultrasound (US)-guided puncture method (US group). The pinch-off grade (POG) was determined using chest X-ray findings following the definition of Hinke, and the progression of POG during the follow-up period of the Landmark and US groups was compared. RESULTS: Sixteen cases in the Landmark group were POG-1 and 3 were POG-2, while all cases in the US group were POG-0 at the time of venipuncture (p < 0.001). Eleven patients in the Landmark group showed some degree of progression of the POG during the follow-up period. In contrast, there were no cases showing progression of the POG in the US group (p = 0.002). CONCLUSIONS: US-guided infraclavicular axillary vein puncture was found to effectively make it possible to avoid POS for the long-term management of TIAPs, as well as at the time of implantation.

Clinical usefulness of mitochondrial transcription factor A expression as a predictive marker in colorectal cancer patients treated with FOLFOX.

We previously reported that mitochondrial transcription factor A (mtTFA) preferentially recognizes cisplatin-damaged DNA via physical interaction with p53 and is upregulated by treatment with cisplatin and fluorouracil (5-FU). The aim of the present study was to evaluate whether expression of mtTFA predicts the clinical outcome in patients with metastatic colorectal cancer treated with modified 5-fluorouracil, leucovorin and oxaliplatin 6 (mFOLFOX6). Fifty-nine patients with metastatic lesions from colorectal cancer treated with mFOLFOX6 were included in this study. The subjects consisted of 25 women and 34 men with a median age of 62 years. The patients were treated with oxaliplatin (85 mg/m(2) ) plus leucovorin (200 mg/m(2) ) as a 2-h infusion on day 1, followed by 5-FU (400 mg/m(2) ) bolus and 46-h continuous infusion of 2400 mg/m(2) . The expressions of mtTFA and p53 of resected primary tumors were examined by immunohistochemical analysis. Of the 59 patients, 33 (complete response 1, partial response 32) achieved a confirmed response to therapy. The positive cytoplasmic staining rate for mtTFA was 44.1% and that for p53 was 59.3%, respectively. Strong expression of mtTFA was detected in eight of 33 complete response/partial response (24.2%) and in 18 of 26 SD/PD (69.2%), indicating that mtTFA expression was significantly correlated with response to chemotherapy (P<0.01). Median overall survival was significantly longer in patients without mtTFA expression (P=0.0493). Multivariate analysis revealed that mtTFA expression significantly affected overall survival (hazard ratio 2.10, P=0.036). Immunohistochemical study of mtTFA may be useful for predicting the clinical outcome of metastatic colorectal cancer patients treated with FOLFOX.

Synthesis and biological evaluation of 4(5)-(6-methylpyridin-2-yl)imidazoles and -pyrazoles as transforming growth factor-beta type 1 receptor kinase inhibitors.

A series of 4(5)-(6-methylpyridin-2-yl)imidazoles 16-19 and -pyrazoles 22-29, 33, and 34 have been synthesized and evaluated for their ALK5 inhibitory activity in an enzyme assay and in cell-based luciferase reporter assays. The 6-quinolinyl imidazole analogs 16 and 18 inhibited ALK5 phosphorylation with IC(50) values of 0.026 and 0.034 microM, respectively. In a luciferase reporter assay using HaCaT cells transiently transfected with p3TP-luc reporter construct, 18 displayed 66% inhibition at 0.05 microM, while competitor compounds 2 and 3 showed 44% inhibition. The binding mode of 18 generated by flexible docking studies with ALK5:18 complex shows that it fits well into the active site cavity of ALK5 by forming broad and tight interactions.

Recombinant human erythropoietin produced in milk of transgenic pigs.

We have developed a line of transgenic swine harboring recombinant human erythropoietin through microinjection into fertilized one cell pig zygotes. Milk from generations F1 and F2 transgenic females was analyzed, and hEPO was detected in milk from all lactating females at concentrations of approximately 877.9+/-92.8 IU/1 ml. The amino acid sequence of rhEPO protein in the transgenic pig milk matched that of commercial rhEPO produced from cultured animal cells. In addition, an F-36 cell line, which proliferates in the presence of hEPO or commercial EPO, was induced to synthesize erythroid by extracts from tg sow milk. This study provides evidence that production of purified rhEPO from transgenic pig milk is a potentially valuable technology, and can be used as a cost-effective alternative in clinical applications as well as providing other clinical advantages.

Late-onset bile leakage after hepatic resection.

BACKGROUND: Postoperative bile leakage can be a serious complication after hepatic resection. Few studies have analyzed patients according to the time of onset of bile leakage. We analyzed differences between patients with early- and late-onset bile leakage after hepatic resection and assessed clinical characteristics and outcomes in patients with late-onset leakage. METHODS: Between 2008 and 2010, 1,009 patients underwent hepatic resection at 4 participating university hospitals and 2 community hospitals. Fifty-two patients (5.1%) with postoperative bile leakage were divided into an early-onset group (<2 weeks after surgery, n = 34) and a late-onset group (≥2 weeks after surgery, n = 18). Patient characteristics and outcomes were collected prospectively and analyzed retrospectively. RESULTS: The proportion of patients who underwent intra-abdominal placement of a drainage catheter was significantly less in the late-onset group than the early-onset group. All 18 patients in the late-onset group developed intra-abdominal infection, and 2 died of sepsis. The proportion of patients who underwent invasive treatment (abdominal paracentesis, endoscopic biliary drainage, or second hepatic resection) was significantly greater in the late-onset group than in the early-onset group. The time to resolution of bile leakage was significantly greater in the late-onset group than the early-onset group. CONCLUSION: Patients should be monitored carefully for bile leakage for several weeks after hepatic resection, because late-onset bile leakage can cause serious complications. Intra-abdominal infection should also be treated as soon as possible, because it may induce refractory bile leakage with serious complications.

Efficacy of preoperative dexamethasone for postoperative nausea and vomiting after laparoscopic cholecystectomy: a large-scale, multicenter, randomized, double-blind, placebo-controlled trial in Japan.

BACKGROUND: To assess the efficacy of preoperative dexamethasone for postoperative nausea and vomiting (PONV) after laparoscopic cholecystectomy (LC) in Japan. METHODS: A total of 270 patients at eight hospitals were randomized to receive dexamethasone 8 mg (n = 136) or placebo (n = 134) intravenously before LC. The primary endpoint was the degree of PONV and antiemetic requirements within 24 h after LC. Secondary endpoints were postoperative complications, postoperative hospital stay, and cost of hospital stay. This study was registered: UMIN-CTR (UMIN000003841). RESULTS: Within 6 h after LC, 17% (23/136) of patients in the dexamethasone group versus 24% (32/134) in the placebo group reported nausea (P = 0.3), and 5% (7/136) versus 7% (10/134) reported vomiting (P = 0.2). Metoclopramide 10 mg was used 0.09 ± 0.31 versus 0.14 ± 0.35 times (P = 0.2). From 6 to 24 h, 10% (14/136) versus 13% (17/134) reported nausea (P = 0.5), and 5% (7/136) versus 5% (7/134) reported vomiting (P = 0.8). Metoclopramide was used 0.04 ± 0.19 versus 0.03 ± 0.17 times (P = 0.8). Postoperative complications and postoperative hospital stay did not differ significantly between the two groups, but the cost of hospital stay was slightly higher in the dexamethasone group (P < 0.05). CONCLUSIONS: Routine use of preoperative dexamethasone for PONV after elective LC in Japan was not shown to have a clinical advantage.

Elevation of CRABP-I in the cerebrospinal fluid of patients with Moyamoya disease.

BACKGROUND AND PURPOSE: The etiology of moyamoya disease (MMD) remains obscure. This study was undertaken to identify specific proteins associated with the pathogenesis of MMD. METHODS: We studied cerebrospinal fluid (CSF) from 20 patients with angiographically confirmed MMD (4 boys and 16 girls; age range, 3 to 13 years; mean, 7.5 years) and 4 control patients with cerebral palsy who underwent selective dorsal rhizotomy (2 boys and 2 girls; age range, 5 to 10 years; mean, 7.3 years). CSF proteins were analyzed by 2-dimensional polyacrylamide gel electrophoresis, and protein identification was performed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The presence of specific CSF protein in patients with MMD was confirmed by Western blotting. In addition, cerebral CSF was also tested in 7 patients who had other brain diseases but no MMD (2 boys and 5 girls; age range, 1 to 12 years; mean, 6.9 years). RESULTS: We identified 1 polypeptide spot (Mr of 13 to 15 kDa and isoelectric point of 5 to 5.5) that was differentially expressed in the CSF samples of MMD patients (mean optical density intensity, 0.36+/-0.24; range, 0.05 to 0.92) and control spinal CSF samples (mean, 0.03+/-0.04; range, 0 to 0.08; P=0.002). This polypeptide was identified as cellular retinoic acid-binding protein (CRABP)-I. High levels of expression of CRABP-I in the CSF from 17 MMD children were confirmed by Western blotting. CONCLUSIONS: The analysis of the CSF of MMD patients reveals high CRABP-I expression. The present study suggests that the elevation of CRABP-I in CSF may be a candidate for pathogenesis of MMD.

An improved process of isomaltooligosaccharide production in kimchi involving the addition of a Leuconostoc starter and sugars.

Isomaltooligosaccharides (IMOs) are α-(1→6)-linked oligodextrans that show a prebiotic effect on Bifidobacterium spp. This study sought to improve IMO synthesis during lactate fermentation in kimchi by inoculating the kimchi fermentation mix with a starter and sugars; the psychrotrophic Leuconostoc citreum KACC 91035 strain with high dextransucrase activity was used as a starter and sucrose (58 mM) and maltose (56 mM) were added as the donor and acceptor for the glucose-transferring reaction of the dextransucrase, respectively. With the addition of both the starter and the sugars and incubation at 10°C, IMOs were produced in kimchi after 3d. Without the starter, the IMO production rate and maximal concentration in kimchi were 15.05 mM/d and 75.27 mM, respectively, whereas with the starter, the rate and concentration increased to 22.04 mM/d and 110.19 mM, respectively. In addition, the sucrose-maltose mix gave an appropriate level of sweetness by releasing fructose and prevented unfavorable polymer synthesis by IMO production. This result suggests that lactic acid bacteria expressing a highly active glycosyltransferase can be used for the synthesis of beneficial oligosaccharides in various fermented foods.

Posterior reversible encephalopathy syndrome in an untreated hypertensive patient after spinal surgery under general anesthesia -A case report-.

Posterior reversible encephalopathy syndrome (PRES) is an unfamiliar term to anesthesiologists, and this is characterized by neurologic symptoms that include mental change, headache, seizure and visual disturbance and also abnormal neuroimaging finding. A 71-year-old female patient was operated on for posterior decompression and total laminectomy under general anesthesia for the spinal stenosis. After the operation, she developed generalized tonic-clonic seizure and a stuporous mentality in the recovery room. The magnetic resonance imaging (MRI) revealed swelling and increased signal intensity at the deep gray nuclei, cerebral cortex and cerebellum. After one week, she returned to an alert mentality and then she was diagnosed with PRES. She was discharged without any neurologic deficit on postoperative day 20. This report describes our experience with PRES after spinal surgery was performed under general anesthesia on a suspected untreated hypertensive patient.

Ramosetron for the prevention of postoperative nausea and vomiting (PONV): a meta-analysis.

BACKGROUND: Postoperative nausea and vomiting (PONV) remains a challenge for patients and health professionals despite various newly developed prophylactic interventions. We reviewed the efficacy and safety of ramosetron in randomized controlled trials (RCTs) for the prevention of PONV. METHODS: We reviewed 18 randomized controlled trials investigating the efficacy and safety of ramosetron in comparison with placebo or any other drugs. Relevant studies were searched in the MEDLINE, SCOPUS, and the Cochrane database libraries. Our end points of concern were prevention of PONV and adverse effects as dichotomous data. RESULTS: The prophylactic effect of 0.3 mg ramosetron was observed in early PON (relative risk, RR: 0.4; 95% CI 0.3-0.6), early POV (RR: 0.3; 95% CI 0.1-0.6), late POV (RR: 0.3; 95% CI 0.1-0.6), but not late PON (RR: 0.7; 95% CI 0.5-1.0). Compared with placebo, the efficacy of 0.3 mg ramosetron in adults and 6 µg/kg in children were consistently beneficial in preventing PONV overall (RR: 0.4; 95% CI: 03-0.6). The effects of 0.3 mg ramosetron and 3 mg granisetron were similar. No serious side effects or adverse events resulted from ramosetron and other active drugs, and incidence was similar to those of the placebo group. CONCLUSIONS: Ramosetron is effective and safe in children and adults without serious adverse effects compared with placebo or other active drugs, as shown in pooled data of RCTs, in terms of the prevention of PONV.

Production of recombinant human von Willebrand factor in the milk of transgenic pigs.

Von Willebrand factor (vWF), a large multimeric glycoprotein present in blood plasma, is a blood protein of the coagulation system. It is defective in von Willebrand disease and is involved in a large number of other diseases, including thrombotic thrombocytopenic purpura-hemolytic uremic syndrome and heyde's syndrome. We have developed a line of transgenic swine harboring recombinant human von Willebrand factor (rhvWF) cDNA through microinjection of fertilized one-cell pig zygotes. Expression of rhvWF in the mammary gland and secretion of rhvWF into the milk of the transgenic swine were confirmed by immunohistochemical and western blot analyses, respectively, and rhvWF proteins were detected in milk from all lactating founder females at concentrations that were 28- to 56-folds greater than that in circulating human plasma. The amino acid sequence of rhvWF protein in the transgenic pig milk matched that of vWF produced from human blood plasma. This study provides evidence that production of rhvWF from transgenic pig milk is a potentially valuable technology and can be used as a cost-effective alternative in clinical applications.

Identification of differentially expressed proteins in a mat1-2-deleted strain of Gibberella zeae, using a comparative proteomics analysis.

Gibberella zeae is a self-fertile ascomycetous fungus that causes important diseases of cereal crops. A comprehensive understanding of sexual reproduction in G. zeae is needed for disease control. To identify fungal proteins involved in this process, we compared the protein profiles of a wild-type strain and its self-sterile strain deleted for MAT1-2, a master regulator of sexual reproduction in G. zeae. Using 2-DE and either MALDI-TOF or ESI-Q-TOF MS, we identified 13 protein spots that showed statistically significant differences in expression levels between the two strains; 11 were reduced and two were increased in abundance in the DeltaMAT1-2 strain. Six of the 13 proteins were similar to those related to cell wall structure and the others were orthologs of proteins involved in metabolism or environmental stress responses. We confirmed that all the genes of the proteins examined were down-regulated during the sexual development stage in the DeltaMAT1-2, DeltaMAT1-1, and other strains deleted for a MAP kinase or a G-protein gene. These data suggest that differences in the protein expression levels are mostly affected by down-regulation of the corresponding genes in the DeltaMAT1-2 strain. To date, this is the first proteomics approach successfully identifying proteins differentially regulated by MAT1-2 in G. zeae.

Antitumor effect to IL-12 administration into the portal vein on murine liver metastasis.

BACKGROUND/PURPOSE: Interleukin (IL)-12 has been shown to possess potent antitumor activity, and an antitumor effect of systemic IL-12 administration has been reported in liver metastasis models. METHODS: In this study, we examined the usefulness of local IL-12 administration into the portal system in the treatment of liver metastasis. First, we confirmed the antitumor effect of recombinant IL-12 (rIL-12) on MC-38 tumors in an intracutaneous model. In the murine liver metastasis model, 1 x 10(5) of MC-38 cells were injected into the portal vein on day 0, and the spleen was transpositioned subcutaneously for administration of rIL-12 continually into the portal system. From days 3 to 7, 0.1 micro g rIL-12 was administered intraperitoneally or intrasplenicly, while Hanks' Balanced Salf Solution (HBSS) was injected intrasplenicly in the control group. RESULTS: The liver weight in the rIL-12 intraperitoneal treatment group (1.88 +/- 0.37 g) and that in the rIL-12 intrasplenic treatment group (1.43 +/- 0.21 g) were significantly less than that in the HBSS group (2.86 +/- 0.74 g; P < 0.05). The numbers of metastatic nodules in the rIL-12 intraperitoneal treatment group (22.3 +/- 17.1) and in the rIL-12 intrasplenic treatment group (12.4 +/- 13.8) were significantly less than that in the HBSS group (137.1 +/- 44.9; P < 0.05). Complete regression of the tumor was observed in one of six mice in the rIL-12 intrasplenic treatment group. This antitumor effect of rIL-12 on MC-38 liver metastasis was not observed in interferon (IFN)-gamma knockout mice. Intraportal administration of IL-12-transduced fibroblasts, which were syngeneic to C57BL/6 mice, had an antitumor effect in the MC-38 liver metastasis model. CONCLUSIONS: These results suggested that the local administration of IL-12 into the portal system would be a useful strategy for the treatment of liver metastasis.