A novel sialyltransferase inhibitor AL10 suppresses invasion and metastasis of lung cancer cells by inhibiting integrin‐mediated signaling

Aberrant sialylation catalyzed by sialyltransferases (STs) is frequently found in cancer cells and is associated with increased cancer metastasis. However, ST inhibitors developed till now are not applicable for clinical use because of their poor cell permeability. In this study, a novel ST inhibitor AL10 derived from the lead compound lithocholic acid identified in our previous study is synthesized and the anti‐cancer effect of this compound is studied. AL10 is cell‐permeable and effectively attenuates total sialylation on cell surface. This inhibitor shows no cytotoxicity but inhibits adhesion, migration, actin polymerization and invasion of α‐2,3‐ST‐overexpressing A549 and CL1.5 human lung cells. Inhibition of adhesion and migration by AL10 is associated with reduced sialylation of various integrin molecules and attenuated activation of the integrin downstream signaling mediator focal adhesion kinase. More importantly, AL10 significantly suppresses experimental lung metastasis in vivo without affecting liver and kidney function of experimental animals as determined by serum biochemical assays. Taken together, AL10 is the first ST inhibitor, which exhibits potent anti‐metastatic activity in vivo and may be useful for clinical cancer treatment. J. Cell. Physiol. 223: 492–499, 2010. © 2010 Wiley‐Liss, Inc.     

Usage of high‐performance mattresses for transport of Indo‐Pacific bottlenose dolphin

Ground transport can be a stressful operation for dolphins if the long period of restraint causes damage to internal organs, especially to the lung, generated by their own weight. Buoyancy is deprived from dolphins under moist transport, in which dolphins are transported on mattresses. Upgrading mattresses is an effective way to modify the transportation method so as to compensate for the loss of buoyancy. In Indo‐Pacific bottlenose dolphins (Tursiops aduncus), we tried to find mattresses that performed well at distributing the dolphins' weight and preserved their pulmonary function. When using EV‐17 (thickness, 50 mm) put on EE‐20 (thickness, 50 mm), a wider support area, less extreme changes in pressure, and lower maximum pressures were observed compared with other mattress systems tested. On this mattress system, lower breathing rates, lower heart rates, and higher exhaled CO₂ concentrations were shown compared with using standard mattresses. These results suggest that the performance of the combination of EV‐17 and EE‐20 is better than that of the standard mattress in terms of the cardiopulmonary function of dolphins. Zoo Biol 27:331–340, 2008. © 2008 Wiley‐Liss, Inc.     

Inhibition of Oncogenic and Activated Wild-Type ras–p21 Protein-Induced Oocyte Maturation by Peptides from the Guanine-Nucleotide Exchange Protein, SOS, Identified from Molecular Dynamics Calculations. Selective Inhibition of Oncogenic ras–p21

In the preceding paper we performed molecular dynamics calculations of the average structures of the SOS protein bound to wild-type and oncogenic ras–p21. Based on these calculations, we have identified four major domains of the SOS protein, consisting of residues 631–641, 676–691, 718–729, and 994–1004, which differ in structure between the two complexes. We have now microinjected synthetic peptides corresponding to each of these domains into Xenopus laevis oocytes either together with oncogenic (Val 12)-p21 or into oocytes subsequently incubated with insulin. We find that the first three peptides inhibit both oncogenic and wild-type p21-induced oocyte maturation, while the last peptide much more strongly inhibits oncogenic p21 protein-induced oocyte maturation. These results suggest that each identified SOS region is involved in ras–stimulated signal transduction and that the 994–1004 domain is involved uniquely with oncogenic ras–p21 signaling.     

Cation exchange chromatography performed in overloaded mode is effective in removing viruses during the manufacturing of monoclonal antibodies

Viral safety is a critical concern with regard to monoclonal antibody (mAb) products produced in mammalian cells such as Chinese hamster ovary cells. Manufacturers are required to ensure the safety of such products by validating the clearance of viruses in downstream purification steps. Cation exchange (CEX) chromatography is widely used in bind/elute mode as a polishing step in mAb purification. However, bind/elute modes require a large volume of expensive resin. To reduce the production cost, the use of CEX chromatography in overloaded mode has recently been investigated. The viral clearance ability in overloaded mode was evaluated using murine leukemia virus (MLV). Even under high-load conditions such as 2,000 g mAb/L resin, MLV was removed from mAb solutions. This viral clearance ability was not significantly affected by resin type or mAb type. The overloaded mode can also remove other types of viruses such as pseudorabies virus and reovirus Type 3 from mAb solutions. Based on these results, this cost-effective overloaded mode is comparable to the bind-elute mode in terms of viral removal.