Heparin affinity high-performance liquid chromatography as an alternative to bioassay of CS23 mutein of recombinant human basic fibroblast growth factor

We investigated whether a chemical assay by high-performance liquid chromatography (HPLC) as an alternative to the complicated and time-consuming bioassay for CS23 mutein of recombinant human basic fibroblast growth factor (rhbFGF-CS23) using the fetal bovine heart endothelial cell line ATCC CRL 1395. Physically, chemically or enzymatically denatured rhbFGF-CS23 was subjected to heparin affinity (HA)-HPLC and the bioassay. Good agreement was observed between the results obtained by these two methods. Moreover, HA-HPLC gave much more reproducible results (RSD = 1.9%, n = 6) than the bioassay (RSD = 7.4%, n = 18). HA-HPLC is therefore a simple, accurate and reproducible alternative to the bioassay for quality control and stability studies for rhbFGF-CS23 preparations. HA-HPLC is also considered to be applicable to assays for FGFs which have heparin affinity and biological activity similar to those of the CS23 mutein.     

Strength and ductility under dynamic loading in fine- grained IN905XL aluminum alloy

Three IN905XL aluminum alloys with fine grain (1 μm), intermediate grain (3 μm), and coarse grain (5 μm) have been developed by a combination of mechanical alloying (MA) and conventional extrusion in order to investigate their mechanical properties at dynamic strain rates of 1 × 10³ and 2 × 10³ s⁻¹ and a quasi-static strain rate of 10^-3 s⁻¹. Flow stresses are found to increase with decreasing grain size for all the strain rates tested. Negative strain-rate sensitivity of flow stress is observed up to 1 × 10³ s⁻¹ in both intermediate- and coarse-grained IN905XL. At the highest strain rate of 2 × 10³ s⁻¹ however, all samples showed a positive strain-rate sensitivity of strength. Total elongation at high strain rates is generally larger than that at low strain rates. Total elongation also decreases with grain size for all the strain rates. This decrease in elongation results from an initiation of microcracks at interfaces between the matrix and particles finely dispersed near grain boundary regions, introduced during MA processing; then, this initiation leads elongation of alloys to small limited values. Formerly with the Department of Mechanical Systems Engineering, University of Osaka Prefecture. This article is based on a presentation made in the symposium “Dynamic Behavior of Materials,” presented at the 1994 Fall Meeting of TMS/ASM in Rosemont, Illinois, October 3-5, 1994, under the auspices of the TMS-SMD Mechanical Metallurgy Committee and the ASM-MSD Flow and Fracture Committee.     

The origination and optimization of Si/SiO/sub 2/ interface roughness and its effect on CMOS performance

As CMOS device dimensions scale down to 100 nm and beyond, the interface roughness between Si and SiO/sub 2/ has become critical to device performance and reliability. Si/SiO/sub 2/ interface roughness degrades channel mobility decreasing drive currents. The authors have used atomic force microscopy to study surface roughness in the processing of 0.16 /spl mu/m CMOS integrated circuits. All of the process steps that could potentially affect the interface roughness have been studied. The results show that oxidation is the major contributor to the interface roughness. The rms roughness is found to be linearly dependent on oxide thickness. Transistors with Si/SiO/sub 2/ interface rms roughness that has been reduced from 1.6 to 1.1 /spl Aring/ by reducing oxide thicknesses show improved device drive currents. This technique for interfacial smoothing and device performance improvement has the advantage of being easily implemented in today's technology.     

Quantitative trait loci affecting 4-nitroquinoline 1-oxide-induced tongue carcinogenesis in the rat

The incidence of tongue carcinomas (TCs) induced by oral administration of 4-nitroquinoline 1-oxide in rats is strain dependent. The inbred Dark-Agouti (DA) strain showed a much higher susceptibility to large mass-forming infiltrative TCs than did the Wistar-Furth (WF) strain. Our previous study (M. Kitano et al, Jpn. J. Cancer Res., 87: 1097-1101, 1996) on crosses between these two strains postulated a dominant susceptibility gene in DA and a dominant resistance gene in WF rats. The present study mapped these loci by analyzing the backcrosses to each parent with simple sequence repeat polymorphisms. Five quantitative parameters were analyzed: (a) the number of TCs > 5 mm in diameter; (b) the total number of TCs per rat; (c) the diameter of the largest TCs (DTCmax values); (d) the number of non-TC cancers per rat; and (e) and the number of cancers of any site per rat. All of these parameters were closely correlated (P < 0.0001). DA rats had a semidominant gene (Stc1) favoring the development of 4-nitroquinoline 1-oxide-induced cancers on chromosome 19, closely linked to D19Mit9. Peak linkage was observed 4 cM distal from D19Mit9, with a logarithm of the odds (lod) score of 5.72 for the number of large TCs and 6.08 for the DTCmax. On the other hand, WF rats had a semidominant gene (Rtc1) mapped between D1Mit1 and D1Mit3, approximately 20 cM from D1Mit1, with a peak lod score of 3.30 for both the number of large TCs and the DTCmax. The main effect of Rtc1 seemed to be to reduce the size of the TCs. The action of these genes was dose dependent and cooperative. The final incidence of TC in DA, WF, F1, and backcross rats seemed to be explained by combinations of genotype at these two loci. Possible candidate genes for Stc1 and Rtc1 are discussed.     

Development of a Cyclogyro-Based Flying Robot With Variable Attack Angle Mechanisms

This paper presents an experimental study on the development of a cyclogyro-based flying robot with a new variable angle of attack mechanism. A cyclogyro is a flying machine supported in the air by power-driven rotors that rotate about a horizontal axis, like the paddle-wheels of a steamboat. Machines of this type have been designed by some companies but there has been no record of any successful flights. Our design starts with a new variable angle of attack mechanism with an eccentric (rotational) point in addition to a rotational point connecting to a motor. The main feature of the mechanism with the eccentric rotational point is the ability to change attack of angles in accordance with the wing positions (as determined by the rotational angles of the cyclogyro) without actuators. The design parameters (wing span, the number of wings, and eccentric distance) of the flying robot are determined through a series of experiments. Experimental results show that the cyclogyro-based flying robot with the new variable angle of attack mechanism is capable of generating sufficient lift force for flying.     

A unified analysis of resonant converters

The general method of analysis for resonant power converters is presented. This analytical method generalizes the idea of state-space-averaging technique to overcome the limitations of the conventional state-space-averaging method. As the result, the characteristics of resonant power converters are clarified so that transfer functions and stability conditions are revealed. In addition, a computer program of analysis based on the proposed method is developed. The program can be applied to various resonant power converters, even when they have parasitic losses and higher-order resonant circuits     

Supramolecular Nanofibers from Collagen-Mimetic Peptides Bearing Various Aromatic Groups at N-Termini via Hierarchical Self-Assembly

Self-assembly of artificial peptides has been widely studied for constructing nanostructured materials, with numerous potential applications in the nanobiotechnology field. Herein, we report the synthesis and hierarchical self-assembly of collagen-mimetic peptides (CMPs) bearing various aromatic groups at the N-termini, including 2-naphthyl, 1-naphtyl, anthracenyl, and pyrenyl groups, into nanofibers. The CMPs (R-(GPO)_n: n > 4) formed a triple helix structure in water at 4 °C, as confirmed via CD analyses, and their conformations were more stable with increasing hydrophobicity of the terminal aromatic group and peptide chain length. The resulting pre-organized triple helical CMPs showed diverse self-assembly into highly ordered nanofibers, reflecting their slight differences in hydrophobic/hydrophilic balance and configuration of aromatic templates. TEM analysis demonstrated that 2Np-CMP_n (n = 6 and 7) and Py-CMP₆ provided well-developed natural collagen-like nanofibers and An-CMP_n (n = 5–7) self-assembled into rod-like micelle fibers. On the other hand, 2Np-CMP₅ and 1Np-CMP₆ were unable to form nanofibers under the same conditions. Furthermore, the Py-CMP₆ nanofiber was found to encapsulate a guest hydrophobic molecule, Nile red, and exhibited unique emission behavior based on the specific nanostructure. In addition to the ability of CMPs to bind small molecules, their controlled self-assembly enables their versatile utilization in drug delivery and wavelength-conversion nanomaterials.     

Effect of oxidized oil on lipogenic enzymes

Male Wistar rats were fed for 4 wk on diets containing 2% oxidized corn oil. Liver tissue was then studied to determine the effect of feeding peroxidized oil on lipogenic enzymes. Although substances which reacted with thiobarbituric acid increased in liver microsomes and mitochondria with increasing peroxide values of the dietary corn oil fed, the activities of glucose-6-phosphate dehydrogenase, malic enzyme and acetyl-CoA carboxylase in liver were unchanged. However, when rats were fed for 2 wk on diets containing 10% fat, of which 0.5, 5 or 10% was unoxidized corn oil and the remainder was hydrogenated beef tallow filler, the lipogenic enzyme activities and also the liver triglyceride levels were observed to decrease with increasing amounts of dietary corn oil. Therefore, although a synthetic diet containing corn oil was easy to oxidize spontaneously, the reductions of lipogenic enzymes in rats fed the diet would not have been caused by lipid peroxides but by unsaturated fatty acids themselves.     

Hydrophobic effects on protein/nucleic acid interaction: enhancement of substrate binding by mutating tyrosine 45 to tryptophan in ribonuclease Tl

Hydrophobic effects on binding of ribonuclease Tl to guaninebases of several ribonucleotides have been proved by mutatinga hydrophobic residue at the recognition site and by measuringthe effect on binding. Mutation of a hydrophobic surface residueto a more hydrophobic residue (Tyr45 – Trp) enhances thebinding to ribonucleotides, including mononucleotide inhibitorand product, and a synthetic substrate-analog trinudeotide aswell as the binding to dinucleotide substrates and RNA. Enhancementson binding to non-substrate ribonucleotides by the mutationhave been observed with free energy changes ranging from –2.2 to – 3 .9 kJ/mol. These changes are in good agreementwith that of substrate binding, –2.3 kJ/mol, which iscalculated from Michaelis constants obtained from kinetic studies.It is shown, by comparing the observed and calculated changesin binding free energy with differences in the observed transferfree energy changes of the amino acid side chains from organicsolvents to water, that the enhancement observed on guaninebinding comes from the difference in the hydrophobic effectsof the side chains of tyrosine and tryptophan. Furthermore,a linear relationship between nucleolytic activities and hydrophobicityof the residues (Ala, Phe, Tyr, Trp) at position 45 is observed.The mutation could not change substantially the base specificityof RNase Tl, which exhibits a prime requirement for guaninebases of substrates.