Lack of PPCA expression only partially coincides with lysosomal storage in galactosialidosis mice: indirect evidence for spatial requirement of the catalytic rather than the protective function of PPCA

Protective protein/cathepsin A (PPCA) is a pleiotropic lysosomal enzyme that complexes with beta-galactosidase and neuraminidase, and possesses serine carboxypeptidase activity. Its deficiency in man results in the neurodegenerative lysosomal storage disorder galactosialidosis (GS). The mouse model of this disease resembles the human early onset phenotype and results in severe nephropathy and ataxia. To understand better the pathophysiology of the disease, we compared the occurrence of lysosomal PPCA mRNA and protein in normal adult mouse tissues with the incidence of lysosomal storage in PPCA(-/-) mice. PPCA expression was markedly variable among different tissues. Most sites that produced both mRNA and protein at high levels in normal mice showed extensive and overt storage in the knockout mice. However, this correlation was not consistent as some cells that normally expressed high levels of PPCA were unaffected in their storage capability in the PPCA(-/-) mice. In addition, some normally low expressing cells accumulated large amounts of undegraded products in the GS mouse. This apparent discrepancy may reflect a requirement for the catalytic rather than the protective function of PPCA and/or the presence of cell-specific substrates in certain cell types. A detailed map showing the cellular distribution of PPCA in nomal mouse tissues as well as the sites of lysosomal storage in deficient mice is critical for accurate assessment of the effects of therapeutic interventions.     

Surface alterations of the bovine oocyte and its investments during and after maturation and fertilization in vitro

Surface characteristics of the bovine oocyte and its investments before, during, and after maturation, and fertilization in vitro were evaluated by scanning electron microscopy (SEM). Oocyte diameters were also measured during SEM analysis of the oocyte. The cumulus cells manifested a compact structure with minimal intercellular spaces among them in the immature oocytes. These became fully expanded with increased intercellular spaces after maturation in vitro, but contracted again after fertilization. The zona pellucida (ZP) showed a fibrous, open mesh-like structure in the maturing and matured oocytes. The size and number of meshes on the ZP decreased dramatically after fertilization. The vitelline surface of immature oocytes was characterized by distribution of tongue-shaped protrusions (TSPs) varying in density. After 10 and 22 hr of maturation incubation, oocyte surface microvilli (MV) increased to become the predominant surface structure, and TSPs decreased substantially. The vitelline surface of fertilized oocytes (at 6 and 20 hr) was similar to that of the matured oocytes, but unfertilized oocytes had less dense MV than did fertilized oocytes (at 20 hr). The diameter of the oocytes decreased from 99 to 80 microns during maturation and increased to 106 microns after insemination (P < 0.05). Membrane maturation was characterized by surface changes from a TSP-predominant pattern to a MV-predominant pattern. Thus, the bovine oocyte maturation process was found to involve the expansion of cumulus cells and the maturation of the ZP, which changes dramatically upon fertilization. Also, volumetric changes occurred in ooplasm processed for SEM following oocyte maturation and insemination.     

Properties of indium oxide/tin oxide multilayered films prepared by ion-beam sputtering

The preparation and characterization of indium oxide (InO _x )/tin oxide (SnO _y ) multilayered films deposited by ion-beam sputtering are described and compared with indium tin oxide (ITO) films. The structure and the optoelectrical properties of the films are studied in relation to the layered structures and the post-deposition annealing. Low-angle X-ray diffraction analysis showed that most films retained the regular layered structures even after annealing at 500° C for 16 h. As an example, we obtained a resistivity of 6×10^–4 cm and a transparency of about 85% in the visible range at a thickness of 110 nm in a multilayered film of InO _x (2.0 nm)/SnO _y (0.2 nm)×50 pairs when annealed at 300° C for 0.5 h in air. Hall coefficient measurements showed that this film had a mobility of 17 cm² V^–1 sec^–1 and a carrier concentration (electron density) of 5×10²⁰ cm^–3.     

Deformation of NiO single crystals below room temperature: dislocation configurations

NiO single crystals prepared by two crystal growth techniques (zone melting in an arc-image furnace and Verneuil crystallization have been deformed by compression along 001 at temperatures as low as 4.2 K, and the dislocation substructure observed by transmission electron microscopy. The Peierls mechanism has been suggested as the mechanism controlling the mechanical behaviour at the lower temperatures. The dislocations generated at cavities found in the zone-melted crystals may be responsible for the increase of the flow stress of these crystals compared with the Verneuil ones.     

Surface tension of liquid3He and4He near the liquid-gas critical point

The surface tension of liquid³He and⁴He was measured near the gas-liquid critical points in the reduced temperature range 3×10^–4–1, where t (T _c –T)/T _c . The critical exponents were found to be ₃=1.289±0.015 for³He and ₄=1.306±0.017 for⁴He. These values are very close to those for classical liquids, and are consistent with the value of 1.28 predicted by Widom, but are apparently different from the exponents previously obtained for liquid helium isotopes, which are near unity. The critical coefficients show good agreement with the quantum-corrected corresponding states theory for the Lennard-Jones 6–12 potential discussed by Young. The interface thickness is deduced from Widom's theory to bed=d ₀t^–v withd ₃₀=0.14±0.03nm and v₃=0.57±0.04 for³He, andd ₄₀=0.37±0.07 nm and v₄=0.58±0.01 for⁴He.     

NANO-MULTILAYERS WITH HIGH PERPENDICULAR ANISOTROPY FOR MAGNETIC RECORDING

(FePt/Ag)n nano-multilayers were deposited on MgO (100) single crystal with laser ablation and then subjected to annealing. FePt L1o grains with (001) texture and thus a large perpendicular magnetic anisotropy constant Ku of the order of 106 J/m3 were formed. A thick Ag layer is found to be favorable for decreasing the dispersion of the easy axis for magnetization. The measurement of time decay of magnetization gave rise to a small activation volume of the order of 10-25m3, showing the promising of being the recording medium for future high density perpendicular recording.     

Age-related accumulation of phosphatidylcholine hydroperoxide in cultured human diploid cells and its prevention by α-tocopherol

The levels of phosphatidylcholine hydroperoxide in serially cultured human fetal diploid fibroblasts at various population doubling levels were determined by high-performance liquid chromatography combined with chemiluminescence detections. This methodology utilizes a mixture of cytochromec and luminol as post-column hydroperoxide group specific luminescent reagents. The cellular hydroperoxide content increased with age from 0.34 to 27.72 pmol/10⁶ cells. At the end of the cells'in vitro lifespan (51st population doubling level), the hydroperoxide content per 10⁶ cells reached about 80 times the level found in cells of the 20th population doubling level. Supplementation of exogenous α-tocopherol to the culture medium prevented hydroperoxide accumulation, but did not extent the lifespanin vitro. The results indicate that substantial intracellular phospholipid hydroperoxide accumulation occurred in the course of aging of human fetal liploid fibroblasts.     

Different feeding and gustatory responses to ecdysone and 20-hydroxyecdysone by larvae of the silkworm,Bombyx mori

The feeding and gustatory responses to ecdysone and 20-hydroxyecdysone were investigated in the silkworm,Bombyx mori. 20-Hydroxyecdysone reduced feeding response strongly in fourth- and fifth-instar larvae, whereas ecdysone had no effect on feeding response. 20-Hydroxyecdysone stimulated the R receptor, the receptor to feeding deterrents, to a great degree. By contrast, ecdysone was much less effective for stimulating the R receptor. These results indicate that ecdysone and 20-hydroxyecdysone have different effects on feeding response due to different interactions with mouthpart chemoreceptors.