Human buprenorphine N-dealkylation is catalyzed by cytochrome P450 3A4

Buprenorphine (BN) is a thebaine derivative with analgesic properties. To identify and characterize the cytochrome P450 (CYP) enzyme(s) involved in BN N-dealkylation, in vitro studies using human liver microsomes and recombinant human CYP enzymes were performed. Norbuprenorphine formation from BN was measured by a simple HPLC-UV assay method, without extraction. The BN N-dealkylation activities in 10 human liver microsomal preparations were strongly correlated with microsomal CYP3A-specific metabolic reactions, i.e. triazolam 1'-hydroxylation (r = 0.954), midazolam 1'-hydroxylation (r = 0.928), and testosterone 6beta-hydroxylation (r = 0.897). Among the eight recombinant CYP enzymes studied (CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A4), only CYP3A4 could catalyze BN N-dealkylation. The apparent KM value for recombinant CYP3A4 was similar to that for human liver microsomes (23.7 vs. 39.3 +/- 9.2 microM). The demonstration of BN N-dealkylation by recombinant CYP3A4 and the agreement in the affinities (apparent KM values) of human liver microsomes and recombinant CYP3A4 provide the most supportive evidence for BN N-dealkylation being catalyzed by CYP3A4.     

Involvement of neutrophil elastase in crescentic glomerulonephritis

To elucidate the role of neutrophils in the tissue damage of crescentic glomerulonephritis (GN), we examined neutrophils infiltrated in renal tissues and the localization of neutrophil elastase (NE), as a neutrophil-derived tissue destructive mediator, using an immunohistochemical technique with antibodies specific for neutrophils and neutrophil elastase; the enzyme histochemical technique (chloroesterase staining) also was used to detect neutrophils. In normal controls, neutrophil infiltration was scarce, and NE was localized in neutrophil cytoplasm. Neutrophils were abundant in crescentic GN and infiltrated in the glomerulus and interstitium; the infiltrating neutrophils were often aggregated. NE was localized in the cytoplasm of neutrophils and also appeared extracellularly (in granular or diffuse patterns) in glomerular necrotizing lesions, crescents, ruptured portions of Bowman's capsules, and in periglomerular and perivascular sites of the interstitium. Moreover, urinary concentration of NE measured by enzyme-linked immunosorbent assay (ELISA) in crescentic GN patients was significantly higher than in normals (93.6 +/- 13.3 v 1.4 +/- 0.5 microg/g x Cr, respectively; P < .001). These data suggest that NE plays a significant role in renal tissue damage, especially in the formation of glomerular necrotizing and crescentic lesions and in periglomerular interstitial lesions of crescentic GN.     

Comparison of biomechanical properties of the incisor periodontal ligament among different species

BACKGROUND: The aim of this study was to obtain a more precise understanding of the mechanical properties of the periodontal ligament in continuously erupting incisors by comparing the shear stress-strain relations among teeth from four closely related species. METHODS: Four species of experimental animals (mice, hamsters, rats, and rabbits) were used. Transverse sections of the left mandibular incisors were cut from the incisal, middle, and basal regions of each incisor. The tooth was pushed out of the alveolar bone in an extrusive direction at 5 mm/min using a materials testing machine. The maximum shear stress, maximum shear strain, tangent modulus, and failure strain energy density were estimated from the resulting stress-strain curve. Polarized light microscopic observations of collagen fibers were also made. RESULTS: All the biomechanical measures tended to decrease from the incisal toward the basal regions in all species. There were large species differences, especially in the incisal region, with the greatest maximum shear stress and failure strain energy density in hamsters. The greatest tangent modulus and the smallest maximum shear strain were observed in mice. The birefringent fiber architectures of the periodontal ligaments in the four species appeared to be similarly organized; the incisal periodontal ligament appeared to have more organized and thicker collagen fibres than did the middle and basal ligaments in the four species. CONCLUSIONS: These results suggest that the regional differences in the biomechanical properties of the periodontal ligament depend upon the developmental stages of the periodontal collagen fibers that may be related to the general arrangement, diameters, and densities of the collagen fiber bundles and the fiber insertions into the alveolar bone and cementum. The species differences in the biomechanical properties may be due to differences in the width of the periodontal ligament and the waviness as well as the strength and stiffness of the periodontal collagen fibers.     

Channel Thickness Dependence of the Magnetic Properties in (Ga,Mn)As FET Structures

We have fabricated a series of field-effect transistor structures with a thin (Ga,Mn)As channel with thickness t of 3.5, 4.0, 4.5, and 5.0 nm, and investigated the effect of electric-field E on their magnetic properties. The Curie temperature T _C showed a clear dependence on the magnitude of E, and its controllable range became larger with decreasing t and reached 15 K for the device with t=3.5 nm, which corresponded to 32% of T _C of the layer.     

Selection of manipulator system for multiple-goal task by evaluating task completion time and cost with computational time constraints

The focus of this study is on the problem of manipulator system selection for a multiple-goal task by evaluating task completion time and cost with computational time constraints. An approach integrating system selection, structural configuration design, layout design, motion planning, and relative cost calculation is proposed to solve this problem within a reasonable computational time. In the proposed approach, multiple-objective particle swarm optimization (MOPSO) is utilized to search for the appropriate manipulator system with appropriate structural configuration from a set of candidate systems. Particle swarm optimization (PSO) and the nearest neighborhood algorithm are employed in layout design and motion planning due to their high convergence speed. Three methods involving a random search algorithm are compared to the proposed approach through a simulation. The simulation is done with a set of tasks and the result shows the effectiveness of the proposed approach.     

Development of silicon wafer packaging technology for deep UV LED

This paper reports a deep‐ultraviolet LED (deep‐UV‐LED) package based on silicon MEMS process technology (Si‐PKG). The package consists of a cavity formed by silicon crystalline anisotropic etching, through‐silicon vias (TSVs) filled with electroplated Cu, bonding metals made of electroplated Ni/AuSn and a quartz lid for hermetic sealing. A deep‐UV LED die is directly mounted in the Si‐PKG by AuSn eutectic bonding without a submount. It has advantages in terms of size, heat dissipation, light utilization efficiency, productivity and cost over conventional AlN ceramic packages. We confirmed a light output of 30 mW and effective reflection on Si (111) cavity slopes in the Si‐PKG. Based on simulation, further improvement of the optical output is expected by optimizing DUV‐LED die mount condition.     

A histochemical method using a substrate of beta-glucuronidase for detection of genetically modified papaya

A histochemical assay for detecting genetically modified (GM) papaya (derived from Line 55-1) is described. GM papaya, currently undergoing a safety assessment in Japan, was developed using a construct that included a beta-glucuronidase (GUS) reporter gene linked to a virus coat protein (CP) gene. Histochemical assay was used to visualize the blue GUS reaction product from transgenic seed embryos. Twelve embryos per fruit were extracted from the papaya seeds using a surgical knife. The embryos were incubated with the substrate 5-bromo-4-chloro-3-indolyl-beta-D-glucuronide (X-Gluc) in a 96-well microtiter plate for 10-15 hours at 37 degrees C. Seventy-five percent of GM papaya embryos should turn blue theoretically. The histochemical assay results were completely consistent with those from a qualitative polymerase chain reaction (PCR) method developed by this laboratory. Furthermore, the method was validated in a five-laboratory study. The method for detection of GM papaya is rapid and simple, and does not require use of specialized equipment.