Purification and partial characterization of 76 kDa transglutaminase in the egg envelope (chorion) of rainbow trout, Oncorhynchus mykiss

Transglutaminase (TGase), responsible for crosslinking between proteins, is known to be localized exclusively in the egg envelope (chorion) of rainbow trout, Oncorhynchus mykiss, and probably participates in the post-fertilization chorion hardening. We purified the TGase from unfertilized egg chorions by sequential chromatography using SP-Sepharose, Q-Sepharose, and TSK-gel G3000SWXL columns. The purified enzyme was a monomeric protein having the molecular mass of 76 kDa. It promoted incorporation of monodansyl-cadaverine into chorion protein and catalyzed the polymerization of chorion subunit proteins. The effect of various reagents suggested that the chorion TGase is a Ca2+-dependent SH-enzyme similar to the well-characterized TGases of various animals. The highest activity was observed at pH 6.0. The amines examined in the present study inhibited the TGase activity of the purified enzyme. However, they did not necessarily cause effective inhibition of its activity. These properties of the chorion TGase were essentially consistent with our previous observations on polymerization of chorion proteins, resulting in chorion hardening. We compared the amino acid composition of the purified TGase with those of the previously characterized TGases of fishes, such as chum salmon and red sea bream. The results suggest that the chorion 76 kDa TGase is not homologous with those liver TGases in terms of amino acid composition.     

ERCP and CT findings of ectopic drainage of the common bile duct into the duodenal bulb

OBJECTIVE: The purpose of this study was to evaluate ERCP and CT findings of ectopic drainage of the common bile duct into the duodenal bulb. CONCLUSION: Although rare, the diagnosis of ectopic drainage of the common bile duct into the duodenal bulb is important to prevent inadvertent damage during biliary tract or gastric surgery and to clarify the cause of chronic peptic ulcers.     

Substrate-induced conformational change in a trimeric ornithine transcarbamoylase

The crystal structure of Escherichia coli ornithine transcarbamoylase (OTCase, EC 2.1.3.3) complexed with the bisubstrate analog N-(phosphonacetyl)-L-ornithine (PALO) has been determined at 2.8-A resolution. This research on the structure of a transcarbamoylase catalytic trimer with a substrate analog bound provides new insights into the linkages between substrate binding, protein-protein interactions, and conformational change. The structure was solved by molecular replacement with the Pseudomonas aeruginosa catabolic OTCase catalytic trimer (Villeret, V., Tricot, C., Stalon, V. & Dideberg, O. (1995) Proc. Natl. Acad. Sci. USA 92, 10762-10766; Protein Data Bank reference pdb 1otc) as the model and refined to a crystallographic R value of 21.3%. Each polypeptide chain folds into two domains, a carbamoyl phosphate binding domain and an L-ornithine binding domain. The bound inhibitor interacts with the side chains and/or backbone atoms of Lys-53, Ser-55, Thr-56, Arg-57, Thr-58, Arg-106, His-133, Asn-167, Asp-231, Met-236, Leu-274, Arg-319 as well as Gln-82 and Lys-86 from an adjacent chain. Comparison with the unligated P. aeruginosa catabolic OTCase structure indicates that binding of the substrate analog results in closure of the two domains of each chain. As in E. coli aspartate transcarbamoylase, the 240s loop undergoes the largest conformational change upon substrate binding. The clinical implications for human OTCase deficiency are discussed.     

Crystallinity variations in kaolinite induced by grinding and pressure treatments

Kaolinite samples treated with uniaxial pressures higher than 0.1 GPa or grinding times greater than 0.3 h show changes in their crystallinity. These changes are easily detected by the X-ray diffraction (XRD) technique and can be quantified from Hinckley (HI), Lietard (R₂) and also reference intensity ratio (RIR) index studies. The sensitivity of these indices to the Crystallinity changes is; HI>RIR>R₂. In the range of pressures and grinding times considered, these values can be diminished by 50%. The variations in these indices with either the pressure or grinding time follow logarithmic laws whose correlation coefficients are closed to unity. Infrared (IR) spectroscopy and thermal analysis studies show low sensitivity to the changes introduced by grinding or pressure. Nevertheless, thermal gravimetric curves confirm the decrease in the dehydroxylation starting point of about 100°C for the pressed and ground samples. Transmission electron microscopy (TEM) reveals important changes in kaolinite particle morphologies after pressure and grinding treatments. The samples compressed at 0.1, 0.32, 0.85, 1.0 and 2.0 GPa show a large number of defects (fractures, bending, deformations and rolling of layers, glide and rotation of the shell). The ground samples show grain boundaries, dislocations, twins and rounded voids. These defects are responsible for the decrease in crystallinity of the kaolinite samples shown in the XRD and IR studies.     

Fast acquisition clock and data recovery circuit with low jitter

This paper presents a half-rate clock and data recovery circuit (CDR)that combines the fast acquisition of a phase selection (PS) delay-locked loop (DLL) with the low jitter of a phase-locked loop (PLL). The PLL acquisition time improves considerably with use of a phase frequency magnitude detector(PFMD) that feeds back an estimate of the magnitude of the frequency difference in addition to the sign. Measurements in 0.5/spl mu/m CMOS technology show operation up to 700 Mb/s, a 7% acquisition range, an initial acquisition time of 8 bit times with jitter of 30% bit time, and jitter of 16 ps after the PLL acquires lock. With a phase frequency detector (PFD), the PLL locks in about 700 ns from an initial frequency difference of 7%. Measurements using a PFMD show the 700 ns PLL acquisition time is reduced on average by about a factor of 5 to 140 ns from an initial 7% frequency difference. The power dissipation is 300mW.     

Corrosion Condition Evaluation at Muddy Creek Bridge

Corrosion rates were measured at the exposed spots of rebars near three corners of Muddy Creek Bridge located in northern West Virginia using a 3LP device. Also, chloride contents of the concrete samples taken from the vicinity of the spots were analyzed using a wet chemical method that had been developed in this study. The average corrosion rate over the three spots was 4.66 mA/m2, which is in the range of corrosion damage possible in 10–15 years. The average chloride content of the concrete over the three spots was 703 parts per million (ppm), which is well above the threshold limit value of 260 ppm. Although the corrosion rate measurement method using a 3LP device is by manual operation, it is found to be reliable and effective, and, thus, recommended for future studies of this kind of research. The wet chemical method developed in this study is found to be effective.     

Floating-patch MEMS antennas on HRS substrate for millimetre-wave applications

Novel floating-patch micro-electro-mechanical system (MEMS) antennas are proposed for millimetre-wave applications. The floating-patch MEMS antennas are fabricated on a high resistivity silicon (HRS) substrate using surface micromachining technology. Simulation and experimental results for reflection coefficients and radiation patterns are presented.     

Integrated Personal Mobility Architecture: A Complete Personal Mobility Solution

The high expectations and demand for users to access the Internet from anywhere at anytime has made user mobility an important part of the design and development of the next generation mobile communications and computing. Traditionally user mobility has been divided into two areas: Terminal Mobility and Personal Mobility. In recent years terminal mobility has focused on the movement of the terminal and developed extensions to IP protocols such as Mobile IP. In contrast, personal mobility has only received limited attention, and is somewhat lagging behind. This research has either focussed on personal mobility in communications or personalisation of operating environments. As a result, to date no framework for providing true personal mobility has emerged. In this paper, we introduce a new personal mobility framework called IPMoA (Integrated Personal Mobility Architecture), which integrates both aspects of personal mobility to provide a complete personal mobility solution, and illustrate the viability of this approach through a proof-of-concept implementation.