Escherichia coli lipoproteins are anchored to either the inner or outer membrane through fatty acyl chains covalently attached to an N-terminal cysteine. Aspartate at position 2 functions to retain lipoproteins in the inner membrane, although the retention is perturbed depending on the residue at position 3. We previously revealed that LolCDE and LolA play critical roles in this lipoprotein sorting. To clarify the sorting signals, the LolA-dependent release of lipoprotein derivatives having various residues at positions 2 and 3 was examined in spheroplasts. When the residue at position 3 was serine, only aspartate at position 2 caused the retention of lipoproteins in spheroplasts. We then examined the release of derivatives having aspartate at position 2 and various residues at position 3. Strong inner membrane retention occurred with a limited number of species of residues at position 3. These residues were present at position 3 of native lipoproteins having aspartate at position 2, whereas residues that inhibited the retention were not. It was also found that a strong inner membrane retention signal having residues other than aspartate at position 2 could be formed through the combination of the residues at positions 2 and 3. These results indicate that the inner membrane localization of native lipoproteins is ensured by the use of a limited number of strong inner membrane retention signals.Various bacteria possess lipoproteins, which have a lipidmodified cysteine at the N terminus. In Escherichia coli, lipoproteins are anchored to the periplasmic side of either the inner or outer membrane through N-terminal fatty acyl chains (1, 2). Lipoproteins are synthesized as precursors in the cytoplasm and then are translocated across the inner membrane, followed by sequential modification reactions leading to the formation of mature lipoproteins on the periplasmic side of the inner membrane (1, 2). Five Lol proteins are involved in the sorting and outer membrane localization of lipoproteins (3-5). LolCDE in the inner membrane releases outer membrane-directed lipoproteins from the inner membrane in an ATP-dependent manner (5, 6), leading to the formation of a complex between the lipoprotein and LolA (3), a periplasmic chaperone. The LolA-lipoprotein complex crosses the periplasm and then interacts with outer membrane receptor LolB, which mediates the anchoring of lipoproteins to the outer membrane (4). The lipoprotein-sorting signal is recognized at the release step, and the inner membrane-specific lipoproteins are not released (3-5).Inouye and collaborators (7) first revealed the importance of aspartate at position 2 for the inner membrane localization of E. coli lipoproteins. They showed that replacement of serine at position 2 of an outer membrane-specific lipoprotein by aspartate caused the protein to remain in the inner membrane. Furthermore, replacement of aspartate at position 2 of an inner membrane-specific lipoprotein by another residue caused outer membrane localization of the protein. Taken together, these re...

The major outer membrane lipoprotein (Lpp) of Escherichia coli requires LolA for its release from the cytoplasmic membrane, and LolB for its localization to the outer membrane. We examined the significance of the LolA-LolB system as to the outer membrane localization of other lipoproteins. All lipoproteins possessing an outer membrane-directed signal at the N-terminal second position were efficiently released from the inner membrane in the presence of LolA. Some lipoproteins were released in the absence of externally added LolA, albeit at a slower rate and to a lesser extent. This LolAindependent release was also strictly dependent on the outer membrane sorting signal. A lipoprotein-LolA complex was formed when the release took place in the presence of LolA, whereas lipoproteins released in the absence of LolA existed as heterogeneous complexes, suggesting that the release and the formation of a complex with LolA are distinct events. The release of LolB, an outer membrane lipoprotein functioning as the receptor for a lipoprotein-LolA complex, occurred with a trace amount of LolA, and therefore was extremely efficient. The LolA-dependent release of lipoproteins was found to be crucial for the specific incorporation of lipoproteins into the outer membrane, whereas lipoproteins released in the absence of LolA were nonspecifically and inefficiently incorporated into the membrane. The outer membrane incorporation of lipoproteins including LolB per se was dependent on LolB in the outer membrane. From these results, we conclude that lipoproteins in E. coli generally utilize the LolA-LolB system for efficient release from the inner membrane and specific localization to the outer membrane.

In vitro studies were carried out with IM-T. This adsorbent consists of polyvinyl alcohol joined with tryptophan side chains and was delivered by ASAHI Medical Co., Ltd., Tokyo/Japan. It was found that IM-T binds immunoglobulins G and M and also immune complexes only moderately but IgE was adsorbed in remarkable amounts. A clear dose-dependent was adsorbed in remarkable amounts. A clear dose-dependent manner of the IgE adsorption could be stated. Kinetic studies revealed that the binding was only slow and gradual.

BackgroundAmputation of an extremity often results in the sensation of a “phantom limb” where the patient feels that the limb that has been amputated is still present. This is frequently accompanied by “phantom limb pain”. We report here the use of milnacipran, a serotonin and norepinephrine reuptake inhibitor, to treat phantom limb pain after amputation of injured or diseased limbs in three patients.Methods and resultsThe severity of phantom pain before and during treatment was quantified using a visual analog scale. In one case, phantom limb pain responded partially to treatment with high doses of paroxetine, and then replacement with milnacipran further improved the pain relief and long-term full pain relief was achieved. In the two other cases, milnacipran was used as first-line treatment and phantom limb pain responded rapidly.ConclusionThese results suggest that milnacipran administration may be useful in phantom limb pain, possibly as a first-line treatment.

Metallic glass is one of the most attractive advanced materials, and many researchers have conducted various developmental research works. Metallic glass is expected to be used as a functional material because of its excellent physical and chemical functions such as high strength and high corrosion resistance. However, the application for small size parts has been carried out only in some industrial fields. In order to widen the industrial application fields, a composite material is preferred for the cost performance. In the coating processes of metallic glass with the conventional deposition techniques, there is a difficulty to form thick coatings due to their low deposition rate. Thermal spraying method is one of the potential candidates to produce metallic glass composites. Metallic glass coatings can be applied to the longer parts and therefore the application field can be widened. The gas tunnel plasma spraying is one of the most important technologies for high quality ceramic coating and synthesizing functional materials. As the gas tunnel type plasma jet is superior to the properties of other conventional type plasma jets, this plasma has great possibilities for various applications in thermal processing. In this study, the gas tunnel type plasma spraying was used to form the metallic glass coatings on the stainless-steel substrate. The microstructure and surface morphology of the metallic glass coatings were examined using Fe-based metallic glass powder and Zr-based metallic glass powder as coating material. For the mechanical properties the Vickers hardness was measured on the cross section of both the coatings and the difference between the powders was compared.