Engineering of Lipid Nanoparticles by the Multifunctionalization of the Surface with Amino Acid Derivatives for the Neutralization of a Target Toxic Peptide

Protein affinity reagents (e.g., antibodies) are often used for basic research, diagnostics, separations, and disease therapy. Although a lot of “synthetic” protein affinity reagents have been developed as a cost-effective alternative to antibodies, their low biocompatibility is a considerable problem for clinical application. Lipid nanoparticles (LNP) represent a highly biocompatible drug delivery agent. However, little has been reported that LNP itself works as a protein affinity reagent in living animals. Here, LNP is engineered for binding to and neutralizing a target toxic peptide in living animals by multifunctionalization with amino acid derivatives. Multifunctionalized LNP (MF-LNP) is prepared using amino acid derivative-conjugated lipids. Optimized MF-LNP exhibits nanomolar affinity to the target toxic peptide and inhibits toxic peptide-dependent hemolysis and cytotoxicity. In addition, MF-LNP captures and neutralizes the toxic peptide after intravenous injection in the bloodstream; in addition, MF-LNP does not release the toxic peptide in the accumulated organ. These results reveal the potential of using LNP as a highly biocompatible protein affinity reagent such as an antidote.     

Cold repeated forming of compact for aluminium foam

A cold repeated forming process of compacts for producing metal foams was developed in order to strongly bond powder particles. In this process, the compact undergoes severe plastic deformation for the strong bonding of particles by repeated backward extrusion and cup compression, and thus the compact largely foams owing to the accumulation of gas released from blowing agents inside the compact during heating. The cold repeated forming process without heating is much simpler than that for the conventional hot extrusion process. The relative density of the foam was decreased by adding silicon powder to the compact, and an aluminium foam having a relative density of 0.27 was obtained using two repeats of backward extrusion and cup compression, 1.5 mass% titanium hydride powder and 4 mass% silicon powder. In addition, a one-piece foam was successively produced from the bonding of two compacts during the foaming in a die. It was found that the cold repeated forming of compacts is effective for the production of metal foams.     

Evaluation of three-dimensional effects in short deep beams using a rigid-body-spring-model

Three-dimensional (3-D) effects in short deep beams without stirrups that failed in shear were investigated experimentally and analytically. Two deep beams with a shear span to depth ratio (a/d) of 0.5 and with different beam widths were tested. The effect of beam width on load-carrying capacity, failure mode, crack pattern and 3-D behavior was investigated, and shape effect due to beam width was clarified. In addition, the beams were analyzed by the 3-D rigid-body-spring model (RBSM). RBSM is a discrete form of modeling that presents realistic behavior from cracking to failure, and 3-D RBSM is applicable to simulate 3-D behavior as well as the confinement effect of concrete. Analytical results in terms of load–displacement curves and crack pattern are compared with the experimental results. Three-dimensional deformations, strut widths and cross-sectional stress distribution are investigated analytically and compared with the experimental results to determine 3-D behavior in detail. The 3-D effects in short deep beams are clarified.     

Drug delivery from nonpeptidic α-amino acid containing polyamides

New nonpeptidic α-amino acid-based polyamides were used for drug delivery. Benzocaine was covalently linked via a spacer to N-protected poly(l-cystyl-l-cystine) and to poly(adipoyl-l-lysine). Its release by α-chymotrypsin attack was followed by UV. It was nearly quantitative within two days with l-cystine-based polyamide. Protected forms of these polyamides were used as matrices. Release of benzocaine entrapped in polyamide from (l-cystine) was controlled by diffusion. When entrapped in polyamide from (l-lysine) it was controlled by diffusion and swelling. Received: 10 January 1997/Accepted: 23 January 1997     

Characteristics of humic substances in the Kuji River waters as determined by high-performance size exclusion chromatography with fluorescence detection

Direct measurement by high-performance size exclusion chromatography with fluorescence detection was applied to the characterization of humic substances in river waters from the Kuji River system, which runs through forest hills and an agricultural plain in Japan. The monitoring wavelength of excitation 320 nm and emission 430 nm corresponds to the fluorescence maxima for aquatic fulvic acid. Chromatograms of the river waters showed four peaks; each peak position was in good agreement among these samples. Peak height ratios for the samples from the upstream Kuji River and its tributaries were different from those of the midstream and downstream sections of the Kuji River, which may reflect differences in the characteristics of humic substances and other organic materials supplied from soil to river.     

Quantum-Mechanical Properties of Protons in Solid Molecular Hydrogen at Megabar Pressures

Quantum distributions of protons in three high-pressure phases of solid molecular hydrogen are investigated by the first-principles path integral molecular dynamics (FP-PIMD) method, in which interatomic forces are calculated precisely based on the density functional theory. The distributions have entirely different symmetries from those predicted by conventional simulation with classical treatment of protons. Especially in phase II, we found that molecular rotation is hindered by quantum fluctuation of protons, having a strong resemblance to a quantum distribution of impurity muonium in crystalline silicon. The mechanism of this “quantum localization” is clarified by a detailed study of the potential energy surface for the molecular rotation.     

Four-base codon-mediated incorporation of non-natural amino acids into proteins in a eukaryotic cell-free translation system

Various four-base codons have been shown to work for the introduction of non-natural amino acids into proteins in an Escherichia coli cell-free translation system. Here, a four-base codon-mediated non-natural mutagenesis was applied to a eukaryotic rabbit reticulocyte cell-free translation system. Mutated streptavidin mRNAs containing four-base codons were prepared and added to a rabbit reticulocyte lysate in the presence of tRNAs that were aminoacylated with a non-natural amino acid and had the corresponding four-base anticodons. A Western blot analysis of translation products indicated that the four-base codons CGGU, CGCU, CCCU, CUCU, CUAU, and GGGU were efficiently decoded by the aminoacyl-tRNAs having the corresponding four-base anticodons. In contrast, the four-base codons AGGU, AGAU, CGAU, UUGU, UCGU, and ACGU were not decoded. The stop codon-derived four-base codons UAGU, UAAU, and UGAU were found to be inefficient, whereas the amber codon UAG and opal codon UGA were efficient for the incorporation of non-natural amino acids. The application of the expanded genetic code in a eukaryotic cell-free system opens the possibility of a four-base codon-mediated incorporation of non-natural amino acids into proteins in living eukaryotic cells.     

The effects of an angiotensin-converting inhibitor (enalapril) on patients with mild cardiac failure--evaluating cardiac function based on the relationship between daily walking pace and heart rate

BACKGROUND: Heart failure has been evaluated by several methods, the New York Heart Association (NYHA) classification of heart failure based on symptoms being used most frequently. However, the degree of heart failure assessed by these criteria does not always correlate with cardiac function in daily life. HYPOTHESIS: The aim of the study was to evaluate cardiac function based on the walking pace/heart rate (HR) relationship to assess the effects of enalapril, an angiotensin-converting enzyme inhibitor, in patients with mild to moderate cardiac function. METHODS: To evaluate cardiac function objectively, we developed a method using a pedometer to count the steps walked while simultaneously recording HR using a Holter electrocardiograph (ECG). Step-count walk rate (WR) was recorded on the magnetic tape of the Holter apparatus, and both HR and walking pace were calculated automatically by the Holter ECG analysis system. Data were determined every hour, and mean pace and HR were plotted along the x and y axes, respectively. The slope of HR x WR was calculated using the least squares method. The slope and the total number of steps were regarded as indicators of cardiac function and quality of life, respectively. We analyzed 36 subjects, consisting of 8 normal volunteers, 8 patients in New York Heart Association (NYHA) class I. 11 in class II, and 9 in class III chronic mild heart failure, during maximal exercise work load by bicycle ergometer; furthermore, fractional shortening of the left ventricle on echocardiogram was determined in 14 patients with chronic mild heart failure and was compared with the slope of HR x WR. Enalapril was administered at a daily dose of 2.5-10 mg for 1-24 months (mean 6 months) in 60 patients to evaluate the effects of this drug on these parameters. RESULTS: There was a significant inverse relationship between maximal work load and the HR x WR slope, and also between the fractional shortening and the slope, suggesting that the slope may reflect the severity of cardiac dysfunction. Furthermore, the slope decreased significantly from 1.8 +/- 1.26 before enalapril to 1.0 +/- 0.94 (mean +/- standard deviation) after drug administration, while the total number of steps increased significantly from 4842 +/- 3581 to 7804 +/- 4793. CONCLUSION: The slope of the graph relating step count and HR proved to be a good, objective indicator of cardiac function, and enalapril therapy improved this parameter.