Isolation and functional characterization of a β-eudesmol synthase, a new sesquiterpene synthase from Zingiber zerumbet Smith

AbstractIn this paper, we have identified a new sesquiterpene synthase gene (ZSS2) from Zingiber zerumbet Smith. Functional expression of ZSS2 in Escherichia coli and in vitro enzyme assay showed that the encoded enzyme catalyzed the formation of β-eudesmol and five additional by-products. Quantitative RT-PCR analysis revealed that ZSS2 transcript accumulation in rhizomes has strong seasonal variations. To further confirm the enzyme activity of ZSS2 and to assess the potential for metabolic engineering of β-eudesmol production, we introduced a gene cluster encoding six enzymes of the mevalonate pathway into E. coli and coexpressed it with ZSS2. When supplemented with mevalonate, the engineered E. coli produced a similar sesquiterpene profile to that produced in the in vitro enzyme assay, and the yield of β-eudesmol reached 100mg/L

Metastatic recurrence in a pancreatic cancer patient derived orthotopic xenograft (PDOX) nude mouse model is inhibited by neoadjuvant chemotherapy in combination with fluorescence-guided surgery with an anti-CA 19-9-conjugated fluorophore.

The aim of this study is to determine the efficacy of neoadjuvant chemotherapy (NAC) with gemcitabine (GEM) in combination with fluorescence-guided surgery (FGS) on a pancreatic cancer patient derived orthotopic xenograft (PDOX) model. A PDOX model was established from a CA19-9-positive, CEA-negative tumor from a patient who had undergone a pancreaticoduodenectomy for pancreatic adenocarcinoma. Mice were randomized to 4 groups: bright light surgery (BLS) only; BLS+NAC; FGS only; and FGS+NAC. An anti-CA19-9 or anti-CEA antibody conjugated to DyLight 650 was administered intravenously via the tail vein of mice with the pancreatic cancer PDOX 24 hours before surgery. The PDOX was brightly labeled with fluorophore-conjugated anti-CA19-9, but not with a fluorophore-conjugated anti-CEA antibody. FGS was performed using the fluorophore-conjugated anti-CA19-9 antibody. FGS had no benefit over BLS to prevent metastatic recurrence. NAC in combination with BLS did not convey an advantage over BLS to prevent metastatic recurrence. However, FGS+NAC significantly reduced the metastatic recurrence frequency to one of 8 mice, compared to FGS only after which metastasis recurred in 6 out of 8 mice, and BLS+NAC with metastatic recurrence in 7 out of 8 mice (p = 0.041). Thus NAC in combination with FGS can reduce or even eliminate metastatic recurrence of pancreatic cancer sensitive to NAC. The present study further emphasizes the power of the PDOX model which enables metastasis to occur and thereby identify the efficacy of NAC in combination with FGS on metastatic recurrence

Efficacy of tumor-targeting Salmonella typhimurium A1-R in combination with anti-angiogenesis therapy on a pancreatic cancer patient-derived orthotopic xenograft (PDOX) and cell line mouse models.

The aim of the present study was to examine the efficacy of tumor-targeting Salmonella typhimurium A1-R treatment following anti-vascular endothelial growth factor (VEGF) therapy on VEGF-positive human pancreatic cancer. A pancreatic cancer patient-derived orthotopic xenograft (PDOX) that was VEGF-positive and an orthotopic VEGF-positive human pancreatic cancer cell line (MiaPaCa-2-GFP) as well as a VEGF-negative cell line (Panc-1) were tested. Nude mice with these tumors were treated with gemcitabine (GEM), bevacizumab (BEV), and S. typhimurium A1-R. BEV/GEM followed by S. typhimurium A1-R significantly reduced tumor weight compared to BEV/GEM treatment alone in the PDOX and MiaPaCa-2 models. Neither treatment was as effective in the VEGF-negative model as in the VEGF-positive models. These results demonstrate that S. typhimurium A1-R following anti-angiogenic therapy is effective on pancreatic cancer including the PDOX model, suggesting its clinical potential

The tumor-educated-macrophage increase of malignancy of human pancreatic cancer is prevented by zoledronic acid.

We previously defined macrophages harvested from the peritoneal cavity of nude mice with subcutaneous human pancreatic tumors as "tumor-educated-macrophages" (Edu) and macrophages harvested from mice without tumors as "naïve-macrophages" (Naïve), and demonstrated that Edu-macrophages promoted tumor growth and metastasis. In this study, Edu- and Naïve-macrophages were compared for their ability to enhance pancreatic cancer malignancy at the cellular level in vitro and in vivo. The inhibitory efficacy of Zoledronic acid (ZA) on Edu-macrophage-enhanced metastasis was also determined. XPA1 human pancreatic cancer cells in Gelfoam co-cultured with Edu-macrophages proliferated to a greater extent compared to XPA1 cells cultured with Naïve-macrophages (P = 0.014). XPA1 cells exposed to conditioned medium harvested from Edu culture significantly increased proliferation (P = 0.016) and had more migration stimulation capability (P<0.001) compared to cultured cancer cells treated with the conditioned medium from Naïve. The mitotic index of the XPA1 cells, expressing GFP in the nucleus and RFP in the cytoplasm, significantly increased in vivo in the presence of Edu- compared to Naïve-macrophages (P = 0.001). Zoledronic acid (ZA) killed both Edu and Naïve in vitro. Edu promoted tumor growth and metastasis in an orthotopic mouse model of the XPA1 human pancreatic cancer cell line. ZA reduced primary tumor growth (P = 0.006) and prevented metastasis (P = 0.025) promoted by Edu-macrophages. These results indicate that ZA inhibits enhanced primary tumor growth and metastasis of human pancreatic cancer induced by Edu-macrophages

Tumor-Targeting Salmonella typhimurium A1-R in Combination with Trastuzumab Eradicates HER-2-Positive Cervical Cancer Cells in Patient-Derived Mouse Models.

We have previously developed mouse models of HER-2-positive cervical cancer. Tumors in nude mice had histological structures similar to the original tumor and were stained by anti-HER-2 antibody in the same pattern as the patient's cancer. We have also previously developed tumor-targeting Salmonella typhimurium A1-R and have demonstrated its efficacy against patient-derived tumor mouse models, both alone and in combination. In the current study, we determined the efficacy of S. typhimurium A1-R in combination with trastuzumab on a patient-cancer nude-mouse model of HER-2 positive cervical cancer. Mice were randomized to 5 groups and treated as follows: (1) no treatment; (2) carboplatinum (30 mg/kg, ip, weekly, 5 weeks); (3) trastuzumab (20 mg/kg, ip, weekly, 5 weeks); (4) S. typhimurium A1-R (5 × 107 CFU/body, ip, weekly, 5 weeks); (5) S. typhimurium A1-R (5 × 107 CFU/body, ip, weekly, 5 weeks) + trastuzumab (20 mg/kg, ip, weekly, 5 weeks). All regimens had significant efficacy compared to the untreated mice. The relative tumor volume of S. typhimurium A1-R + trastuzumab-treated mice was smaller compared to trastuzumab alone (p = 0.007) and S. typhimurium A1-R alone (p = 0.039). No significant body weight loss was found compared to the no treatment group except for carboplatinum-treated mice (p = 0.021). Upon histological examination, viable tumor cells were not detected, and replaced by stromal cells in the tumors treated with S. typhimurium A1-R + trastuzumab. The results of the present study suggest that S. typhimurium A1-R and trastuzumab in combination are highly effective against HER-2-expressing cervical cancer

Oxygen reduction at negatively charged iron porphyrins heat-treated and bridged by alkaline-earth metal ions

金沢大学理工研究域物質化学系An ordered network of tetrasodium tetra(4-sulphonatophenyl)porphyrin iron(III) chloride (FeTPPS4Na4), which exhibited a higher catalytic activity for oxygen reduction than Co and Ir(CO)TPPS 4Na4, was fabricated by complexation with alkaline-earth metal ions. Heat treatment of these complexes enhanced their catalytic activity with the highest performance observed with Ba2+-FeTPPS4. The onset potential for oxygen reduction (Eonset) was 720mVvs. Ag|AgCl, which is almost the same as that for Pt-impregnated carbon black. The number of electrons, n, transferred during oxygen reduction at a Ba2+- FeTPPS4-coated electrode, as determined by rotating ring-disc experiments, was 3.9 and suggests that oxygen was reduced to water. A neutral solution of FeTPPS4Na4 was acidified by the addition of barium ion, and the elemental ratio Fe:S:Ba in the resulting Ba 2+-FeTPPS4 complex was approximately 1:4:2.5. This suggests the formation of a highly ordered network with Fe sites bridged with barium ions in addition to the normal salt bridges between sulphonates. Catalytic oxidation reactions with nitric oxide and nitrite ion as the intermediate at the heat-treated Ba2+-FeTPPS4 indicate the formation of Fe(IV) and Fe(IV) π-cation radicals or Fe(V). The agreement between the potential of Fe(IV) π-cation radical formation and E onset suggests that the redox cycle of the Fe(III)/Fe(IV) π-cation radical provides a sufficient driving force for the observed 4-electron reduction of oxygen at the heat-treated Ba2+-FeTPPS4 electrode. © 2010 Elsevier Ltd. All rights reserved

Study on reduction of residual stress induced during rapid tooling process: Influence of heating conditions on residual stress

金沢大学理工研究域機械工学系This paper deals with the reduction of residual stress induced during the selective laser melting with a mixture of ferrous based metal powder. To evaluate the residual stresses induced during layered manufacturing processes, a strain gauge is attached on the bottom face of the base plate. The residual stress within the consolidated structure is calculated from the amount of strain change measured by the strain gauge when the consolidated structure is cut with an end mill. The influences of base plate thickness and consolidated structure height on the residual stresses are investigated. In addition, the effect of pre-heating and post heating by a laser beam irradiation are evaluated. The results showed that the deformation of the base plate increased with the increase of the consolidated layer and the decrease of base plate thickness, and the deformation was flattened when the consolidated structure was completely removed with the end mill. The deformation was related to the induced residual stresses. The residual stress distribution within the consolidated structure in the z direction was extremely large at the top layer of the structure and the boundary between the base plate and consolidated structure. The residual stress at the first layer of the structure decreased when the base plate was heated before consolidating the deposited powder. The residual stresses decreased when each of the consolidated layers was repeatedly heated by the laser beam irradiation. © (2010) Trans Tech Publications

Study on laser assisted milling of ferrous based consolidated material

金沢大学理工研究域機械工学系This study deals with the laser beam assisted milling to improve the machinability of a consolidated structure formed by layered manufacturing processes. The laser beam used is a continuous CO2 laser with a maximum output power of 100 W. The metal powder for layered manufacturing is a ferrous-based mixture. To investigate the influence of laser conditions on the machinability, specific cutting force, tool wear and tool temperature of the flank face are measured. The results showed that the specific cutting force of the consolidated structure was decreased by the effect of energy input on the structure surface. The progress of wear on the flank face assisted by the laser beam was 20% slower due to the softening of the consolidated structure and the melting of the powder which remained on the structure surface. The dimensional accuracy and surface finishing of the consolidated structure were influenced by the rise of the tool temperature on the flank face. © KSME & Springer 2010