Membrane-Type 1 Matrix Metalloproteinase Cleaves Cd44 and Promotes Cell Migration

Migratory cells including invasive tumor cells frequently express CD44, a major receptor for hyaluronan and membrane-type 1 matrix metalloproteinase (MT1-MMP) that degrades extracellular matrix at the pericellular region. In this study, we demonstrate that MT1-MMP acts as a processing enzyme for CD44H, releasing it into the medium as a soluble 70-kD fragment. Furthermore, this processing event stimulates cell motility; however, expression of either CD44H or MT1-MMP alone did not stimulate cell motility. Coexpression of MT1-MMP and mutant CD44H lacking the MT1-MMP–processing site did not result in shedding and did not promote cell migration, suggesting that the processing of CD44H by MT1-MMP is critical in the migratory stimulation. Moreover, expression of the mutant CD44H inhibited the cell migration promoted by CD44H and MT1-MMP in a dominant-negative manner. The pancreatic tumor cell line, MIA PaCa-2, was found to shed the 70-kD CD44H fragment in a MT1-MMP–dependent manner. Expression of the mutant CD44H in the cells as well as MMP inhibitor treatment effectively inhibited the migration, suggesting that MIA PaCa-2 cells indeed use the CD44H and MT1-MMP as migratory devices. These findings revealed a novel interaction of the two molecules that have each been implicated in tumor cell migration and invasion

Improving the efficiency of essential-oil extraction from Abies sachalinensis with an underwater shockwave pretreatment

Abies sachalinensis (Sakhalin fir) is a conifer species belonging to the family Pinaceae that is native to and widely distributed throughout Sakhalin Island, the southern Kurils (Russia), and northern Hokkaido (Japan). The essential oil of A. sachalinensis has been found to be an active removal agent, similar to γ-terpinene, myrcene, and β-phellandrene, which effectively remove nitrogen dioxide. Essential oils provide a relaxing effect; the use of essential oils is expected to improve overall air quality.                Underwater shockwaves generate instantaneous high pressure that reaches the entire cell and causes multiple cracks along the tracheids, causing the pit membrane to flake off through spalling destruction. These cracks function as permeation pathways [1]; this application was expected to result in a more effective essential oil extraction by subsequent steam distillation [2]. We, herein, introduce a novel application of this pretreatment process aimed at improving the efficiency of essential-oil extraction from A. sachalinensis leaves and branches. A. sachalinensis leaves and branches were oven-dried (40-45 °C) to a moisture content of 10% or less, and were subjected to the shockwave pretreatment or left untreated before essential-oil extraction by steam distillation. Chemical analysis was performed using gas chromatography-mass spectrometry. The essential-oil yields of raw untreated and untreated dried leaves were 5.1 and 2.4 g/kg of leaf dry weight (DW), respectively. Upon application of a 3.0 kV, 3.6 kJ shockwave, the essential-oil yield increased with the number of shockwave cycles; the yield was 32.7 g/kg DW after 10 cycles, a 13.6-fold increase compared to that of the untreated dried leaves. In addition, sesquiterpenes increased by more than 30-fold in content compared to that of untreated dried leaves. Thus, these results suggest that instantaneous high-pressure treatment, as a pretreatment for conventional steam distillation, has a distinct advantage in increasing the essential-oil yield and extracting the bioactive components. Furthermore, this method also can be used for the pretreatment of microwave essential-oil extraction or steam distillation under reduced pressure

Phase I clinical trial of autologous NK cell therapy using novel expansion method in patients with advanced digestive cancer

Table S2. Change in NK population in PBL

Co-transfection of EYFP-GH and ECFP-rab3B in an experimental pituitary GH3 cell: a role of rab3B in secretion of GH through porosome.

Recently, in order to elucidate the role of rab3B in porosome, we have observed the incorporation of rab3B in the secretion of GH through porosome under confocal laser scanning microscopy (CLSM). Transfected cells with GH-EYFP fusion protein and rab3B-ECFP fusion protein were observed under CLSM, which showed the colocalization of EYFP-GH and ECFP-rab3B in the budding configuration of secretory process. These structural and functional images of rab3B imply the incorporation of rab3B in the secretion of GH through porosome

Multi-Wavelength Photometric and Polarimetric Observations of the Outburst of 3C 454.3 in Dec. 2009

In December 2009, the bright blazar, 3C 454.3 exhibited a strong outburst in the optical, X-ray and gamma-ray regions. We performed photometric and polarimetric monitoring of this outburst in the optical and near-infrared bands with TRISPEC and HOWPol attached to the Kanata telescope. We also observed this outburst in the infrared band with AKARI, and the radio band with the 32-m radio telescope of Yamaguchi University. The object was in an active state from JD 2455055 to 2455159. It was 1.3 mag brighter than its quiescent state before JD 2455055 in the optical band. After the end of the active state in JD 2455159, a prominent outburst was observed in all wavelengths. The outburst continued for two months. Our optical and nearinfrared polarimetric observations revealed that the position angle of the polarization (PA) apparently rotated clockwise by 240 degrees within 11 d in the active state (JD 2455063-2455074), and after this rotation, PA remained almost constant during our monitoring. In the outburst state, PA smoothly rotated counterclockwise by 350 degrees within 35 d (JD 2455157-2455192). Thus, we detected two distinct rotation events of polarization vector in opposite directions. We discuss these two events compared with the past rotation events observed in 2005, 2007 and 2008.Comment: 9 pages, 6 figures, accepted for publication in PAS

Inhibitory effects of local anesthetics on the proteasome and their biological actions

Local anesthetics (LAs) inhibit endoplasmic reticulum-associated protein degradation, however the mechanisms remain elusive. Here, we show that the clinically used LAs pilsicainide and lidocaine bind directly to the 20S proteasome and inhibit its activity. Molecular dynamic calculation indicated that these LAs were bound to the β5 subunit of the 20S proteasome, and not to the other active subunits, β1 and β2. Consistently, pilsicainide inhibited only chymotrypsin-like activity, whereas it did not inhibit the caspase-like and trypsin-like activities. In addition, we confirmed that the aromatic ring of these LAs was critical for inhibiting the proteasome. These LAs stabilized p53 and suppressed proliferation of p53-positive but not of p53-negative cancer cells

A novel transgenic chimaeric mouse system for the rapid functional evaluation of genes encoding secreted proteins

A major challenge of the post-genomic era is the functional characterization of anonymous open reading frames (ORFs) identified by the Human Genome Project. In this context, there is a strong requirement for the development of technologies that enhance our ability to analyze gene functions at the level of the whole organism. Here, we describe a rapid and efficient procedure to generate transgenic chimaeric mice that continuously secrete a foreign protein into the systemic circulation. The transgene units were inserted into the genomic site adjacent to the endogenous immunoglobulin (Ig) κ locus by homologous recombination, using a modified mouse embryonic stem (ES) cell line that exhibits a high frequency of homologous recombination at the Igκ region. The resultant ES clones were injected into embryos derived from a B-cell-deficient host strain, thus producing chimaerism-independent, B-cell-specific transgene expression. This feature of the system eliminates the time-consuming breeding typically implemented in standard transgenic strategies and allows for evaluating the effect of ectopic transgene expression directly in the resulting chimaeric mice. To demonstrate the utility of this system we showed high-level protein expression in the sera and severe phenotypes in human EPO (hEPO) and murine thrombopoietin (mTPO) transgenic chimaeras

Prevention of hypoglycemia by intermittent-scanning continuous glucose monitoring device combined with structured education in patients with type 1 diabetes mellitus : A randomized, crossover trial

Aims: We conducted a randomized, crossover trial to compare intermittent-scanning continuous glucose monitoring (isCGM) device with structured education (Intervention) to self-monitoring of blood glucose (SMBG) (Control) in the reduction of time below range. Methods: This crossover trial involved 104 adults with type 1 diabetes mellitus (T1DM) using multiple daily injections. Participants were randomly allocated to either sequence Intervention/Control or sequence Control/Intervention. During the Intervention period which lasted 84 days, participants used the first-generation FreeStyle Libre (Abbott Diabetes Care, Alameda, CA, USA) and received structured education on how to prevent hypoglycemia based on the trend arrow and by frequent sensor scanning (≥10 times a day). Confirmatory SMBG was conducted before dosing insulin. The Control period lasted 84 days. The primary endpoint was the decrease in the time below range (TBR; <70 mg/dL). Results: The time below range was significantly reduced in the Intervention arm compared to the Control arm (2.42 ± 1.68 h/day [10.1 %±7.0 %] vs 3.10 ± 2.28 h/day [12.9 %±9.5 %], P = 0.012). The ratio of high-risk participants with low blood glucose index >5 was significantly reduced (8.6 % vs 23.7 %, P < 0.001). Conclusions: The use of isCGM combined with structured education significantly reduced the time below range in patients with T1DM

Characterization of pullulanase (PUL)-deficient mutants of rice (Oryza sativa L.) and the function of PUL on starch biosynthesis in the developing rice endosperm

Rice (Oryza sativa) allelic sugary1 (sug1) mutants defective in isoamylase 1 (ISA1) accumulate varying levels of starch and phytoglycogen in their endosperm, and the activity of a pullulanase-type of a debranching enzyme (PUL) was found to correlate closely with the severity of the sug1 phenotype. Thus, three PUL-deficient mutants were generated to investigate the function of PUL in starch biosynthesis. The reduction of PUL activity had no pleiotropic effects on the other enzymes involved in starch biosynthesis. The short chains (DP ≤13) of amylopectin in PUL mutants were increased compared with that of the wild type, but the extent of the changes was much smaller than that of sug1 mutants. The α-glucan composition [amylose, amylopectin, water-soluble polysaccharide (WSP)] and the structure of the starch components (amylose and amylopectin) of the PUL mutants were essentially the same, although the average chain length of the B2-3 chains of amylopectin in the PUL mutant was ∼3 residues longer than that of the wild type. The double mutants between the PUL-null and mild sug1 mutants still retained starch in the outer layer of endosperm tissue, while the amounts of WSP and short chains (DP ≤7) of amylopectin were higher than those of the sug1 mutant; this indicates that the PUL function partially overlaps with that of ISA1 and its deficiency has a much smaller effect on the synthesis of amylopectin than ISA1 deficiency and the variation of the sug1 phenotype is not significantly dependent on the PUL activities