Picoliter volume glass tube array fabricated by Si electrochemical etching process

Fabrication process for picoliter volume SiO₂ glass tube array partially embedded in Si wafer was developed. As a template for the glass tubes, macropore array was formed at the surface of n-Si(1 0 0) wafer by photo-assisted electrochemical etching process. The area-selective formation of the array was achieved by applying Au/Cr micropatterns formed at the back-side surface of the substrate as the shade mask, which controls the illumination condition to optimize the etching reaction conditions. Subsequently, surface of the macropores was wet-thermally oxidized to form glass layer, and the bulk Si region was removed by alkaline etching, remaining the “glass tubes”. As a result of complete removal of the bulk Si, released glass tubes were obtained. By partial removal of the bulk Si part, the glass tubes were exposed, fixed in the remaining Si substrate in the form of well-ordered array. It was confirmed that the depth, the exposed region and the wall thickness of each glass tube were controllable by adjusting the parameters such as the duration of the Si electrochemical etching, the alkaline etching and the wet-thermal oxidation, respectively. In order to demonstrate microreaction in the glass tube, aqueous rhodamine B solution was injected into the tubes and excitation light was irradiated to them. As a result, the fluorescence of rhodamine B was clearly detected, confirming the applicability of the glass tubes for various kinds of devices and systems such as microreactors.     

Engineering of alpha1-antitrypsin variants selective for subtilisin-like proprotein convertases PACE4 and PC6: importance of the P2' residue in stable complex formation of the serpin with proprotein convertase

Furin and PACE4, members of the subtilisin-like proprotein convertase (SPC) family, have been implicated in the metastatic progression of certain tumors in addition to the activation of viral coat proteins and bacterial toxins, indicating that these enzymes are potential targets for therapeutic agents. Alpha1-Antitrypsin Portland is an engineered alpha1-antitrypsin designed as a furin-specific inhibitor and has been used as a tool in the functional analysis of furin. In this work, we engineered rat alpha1-antitrypsin to create a PACE4-specific inhibitor. Substituting Arg-Arg-Arg-Arg for Ala-Val-Pro-Met(352) at P4-P1 and Ala for Leu(354) at P2' created a potent PACE4- and PC6-specific inhibitor. This variant (RRRRSA) formed an SDS- and heat-stable serpin/proteinase complex with PACE4 or PC6 and inhibited both enzyme activities. The RRRRSA variant was efficiently cleaved by furin without formation of the stable complex. This is the first report of a highly selective protein-based inhibitor of PACE4 and PC6. This inhibitor will be useful in delineating the roles of PACE4 and PC6 localized in the extracellular matrix.     

Detection of irradiated meat, fish and their products by measuring 2-alkylcyclobutanones levels after frozen storage

2-Alkylcyclobutanones, such as 2-dodecylcyclobutanone and 2-tetradecylcyclobutanone, were analyzed to assess the irradiation history of irradiated meats or fish, and cooked foods with irradiated ingredients, which had been stored frozen for up to one year. The purpose of the study was to show that irradiated meats could be detected even after having been stored in the distribution system. 2-Alkylcyclobutanones showed a small decrease in irradiated raw meats that had been stored frozen for one year. Cooked foods, such as pancake and fried chicken made with irradiated eggs and chicken, respectively, contained detectable levels of 2-alkylcyclobutanones after storage frozen for one year. The 2-alkylcyclobutanones became undetectable in highly dried samples, such as feed for lab animals, during the same period.     

Fabrication of magnetic nanodot arrays for patterned magnetic recording media

Fabrication processes of arrayed magnetic nanodots for the use of patterned magnetic recording media were reviewed. One candidate for the patterned media is ordered assemble of magnetic nanoparticles, and the other is patterned magnetic thin films fabricated using various micro/nano scale machining processes. For the formation of patterned masks and molds, lithography processes as well as self-organized pattern formation are utilized. For the deposition processes of magnetic dots, electrochemical deposition processes were widely used. These fabrication processes are reviewed mainly from recent reports. The recording systems for the patterned media including probe-type-recording are also overviewed.     

Possible Roles of Calreticulin in Uterine Decidualization and Receptivity in Rats and Humans

Previous in vitro studies have suggested that calreticulin (CALR), which is responsible for the folding and quality control of glycoproteins, may be associated with decidualization. However, its precise role in regulating decidualization has not been explored in vivo. Here, we used pregnant rat models to examine endometrial CALR expression during the peri-implantation period. We also examined whether polypectomy, a procedure that could ameliorate infertility, alters the endometrial expression levels of CALR and several implantation factors in women diagnosed as infertile. In rats, uterine CALR was expressed at a high level at the implantation site, and a marked increase in CALR expression was observed in decidual cells of normal pregnancy. In addition, endometrial CALR expression was enhanced by either administration of estradiol-17β in the delayed implantation rat model or the artificial induction of decidualization in the pseudopregnant rat. In cultured stromal cells, siRNA-mediated silencing of CALR inhibited the decidual stimulus-induced expression of prolactin, decidual/trophoblast prolactin-related protein, and connexin 43. In humans, the endometrial expression levels of the mRNAs encoding CALR and the implantation-related factor insulin-like growth factor binding protein (IGFBP)-7 tended to increase after polypectomy. The strongest positive correlation between expression levels before polypectomy was observed for IGFBP-7 and CALR, and the strength of this correlation increased after the surgery. Thus, endometrial CALR may play a role in the formation of decidua, and the polypectomy of infertile patients may result in the co-operative expression of endometrial factors, including CALR, that could enhance endometrial receptivity.     

Constrained neural network learning algorithm for estimating advertising effectiveness

This paper proposes a neural network algorithm for estimating advertising effectiveness. The basic premise under this algorithm is that the relationship between input and output monotonically increases. The algorithm is useful for domains such as estimation of advertising effectiveness, for which only deficient data are available. The experimental simulation shows that the algorithm builds a reasonable model for estimating advertising effectiveness. © 1998 Scripta Techinca, Electr Eng Jpn, 126(1): 34–41, 1999     

Preparation and mechanical properties of polystyrene–clay hybrids

Polystyrene–clay hybrids (PSCHs) were prepared by melt blending a styrene vinyloxazoline copolymer with organophilic clay. In the PSCHs, the silicate layers of the clay were delaminated and dispersed homogeneously to the nanometer level. The moduli of the PSCHs were higher than that of the PS copolymer. For example, the tensile modulus of the PSCH with 5 wt % clay was 1.4 times higher compared to that of the PS copolymer. © 1999 John Wiley & Sons, Inc. J Appl Polym Sci 74: 3359–3364, 1999     

Development of a quantitative real-time PCR method to enumerate total bacterial counts in ready-to-eat fruits and vegetables

A newly developed real-time PCR assay rapidly quantifies the total bacterial numbers in contaminated ready-to-eat vegetables and fruits compared with the standard plate count method. Primers targeting the rpoB gene, which encodes for the beta subunit of the bacterial RNA polymerase and which is common to most bacterial species, was used instead of the 16S rRNA gene, which has multiple copies and varies among bacterial species. A primer pair specific for rpoB was confirmed to amplify rpoB in a wide range of bacterial species after we assessed 49 strains isolated from five kinds of fruits and vegetables. We purchased fruits and vegetables from retail shops and enumerated the bacteria associated with them by use of real-time PCR and compared this to the number found by the culture method. We found a high correlation between the threshold PCR cycle number when compared with the plate count culture number. The real-time PCR assay developed in this study can enumerate the dominant bacterial species in ready-to-eat fruits and vegetables.     

Physician-received scatter radiation with angiography systems used for interventional radiology: comparison among many X-ray systems

Radiation protection for interventional radiology (IR) physicians is very important. Current IR X-ray systems tend to use flat-panel detectors (FPDs) rather than image intensifiers (IIs). The purpose of this study is to test the hypothesis that there is no difference in physician-received scatter radiation (PRSR) between FPD systems and II systems. This study examined 20 X-ray systems in 15 cardiac catheterisation laboratories (11 used a FPD and 9 used an II). The PRSR with digital cineangiography and fluoroscopy were compared among the 20 X-ray systems using a phantom and a solid-state-detector electronic pocket dosemeter. The maximum PRSR exceeded the minimum PRSR by ~12-fold for cineangiography and ~9-fold for fluoroscopy. For both fluoroscopy and digital cineangiography, the PRSR had a statistically significant positive correlation with the entrance surface dose (fluoroscopy, r = 0.87; cineangiography, r = 0.86). There was no statistically significant difference between the average PRSR of FPDs and IIs during either digital cineangiography or fluoroscopy. There is a wide range of PRSR among the radiography systems evaluated. The PRSR correlated well with the entrance surface dose of the phantom in 20 X-ray units used for IR. Hence, decreasing the dose to the patient will also decrease the dose to staff.     

Distributions of density and fission products in the reaction product between irradiated MOX fuel and molten zircaloy-2

Two- and three-dimensional images were obtained by X-ray CT in the reaction product between zircaloy-2 cladding tube and MOX fuel. The gamma-ray intensity distributions in the same specimen were also obtained by gamma-ray measurements of two fission products (Cs-137 and Eu-154) and one neutron-activated nuclide (Co-60). The average values of the fuel density (about 10.5 g/cm³) and the cladding density (about 6.55 g/cm³) were obtained in the metallic phase region by evaluation of the density distributions on two-dimensional X-ray CT images. The distributions of the crushed fuel pellet and the pores were also clearly observed in the three-dimensional X-ray CT images. The following results were found from the gamma-ray measurement. First, Cs-137 was observed in the unreacted fuel region and the pore region in the metallic phase region. Second, Eu-154 was widely distributed to all regions. Finally, Co-60 was confirmed only in the metallic phase region.