Membrane topology of the Rickettsia prowazekii ATP/ADP translocase revealed by novel dual pho-lac reporters

Here, we report the construction and characterization of dual reporters, consisting of both an Escherichia coli alkaline phosphatase (AP) gene and an alpha-fragment of the beta-galactosidase (BG) gene, for studying membrane protein topology by the gene fusion approach. Each of the reporters, when fused to periplasmic domains of polytopic proteins, produces fusions with high AP activity and, when fused to cytoplasmic domains, produces fusions with high BG activity in E. coli strains capable of alpha-complementation. The dual nature of these reporters simplifies interpretation of data obtained with poorly expressed fusions and allows one to evaluate the reliability of topological data. Deleterious effects resulting from the cell's attempt to export the full-length BG are eliminated in this approach. We describe dual indicator plates that allow for discrimination between colonies bearing cytoplasmic fusions, periplasmic fusions, and no fusions. We have generated a set of fusions to the topologically well-studied lactose permease of E. coli and demonstrated that topological information generated by these new reporters is in good agreement with the existing model. We used this new methodology for the determination of membrane topology of the Rickettsia prowazekii ATP/ADP translocase (Tlc). Our results were in agreement with the proposed in silico topological model in which Tlc traverses the cytoplasmic membrane of E. coli 12 times with its N and C termini facing the cytoplasm.     

Design of micro-retainer to counter-balance spinning seismic proof mass

An innovative magnetic module which is concurrently capable of performing as an actuator and a sensor is proposed and analyzed. The magnetic module is basically similar to a micro-scale Linear Variable Differential Transformer (LVDT) which is employed to actively adjust the height of a rotating seismic disc used in a gyroscope so that the pitch rate induced by Coriolis effect, due to exerted angular excitation, can be accurately measured. That is, the micro-magnetic module acts like an actuator for height regulation on the seismic proof mass by applying appropriate electric current to the primary winding of the LVDT unit. On the other hand, the secondary winding pair of the LVDT acts like a sensor which can detect the gap change, between the seismic rotating disc and the micro-magnetic LVDT module, and the rotation speed (i.e., frequency) by the induced current.     

钢坯连续加热炉计算机在线控制数学模型

在离线模型的基础上.开发了加热炉在线计算机控制数学模型;这种模型系统不仅能够最大程度地节能,而且保证了出钢温度的准确控制,结果表明:出炉钢坯温度与目标出炉温度偏差小于15℃.     

Efficient finite-difference time-domain scheme for light scattering by dielectric particles: application to aerosols

We have examined the Maxwell-Garnett, inverted Maxwell-Garnett, and Bruggeman rules for evaluation of the mean permittivity involving partially empty cells at particle surface in conjunction with the finite-difference time-domain (FDTD) computation. Sensitivity studies show that the inverted Maxwell-Garnett rule is the most effective in reducing the staircasing effect. The discontinuity of permittivity at the interface of free space and the particle medium can be minimized by use of an effective permittivity at the cell edges determined by the average of the permittivity values associated with adjacent cells. The efficiency of the FDTD computational program is further improved by use of a perfectly matched layer absorbing boundary condition and the appropriate coding technique. The accuracy of the FDTD method is assessed on the basis of a comparison of the FDTD and the Mie calculations for ice spheres. This program is then applied to light scattering by convex and concave aerosol particles. Comparisons of the scattering phase function for these types of aerosol with those for spheres and spheroids show substantial differences in backscattering directions. Finally, we illustrate that the FDTD method is robust and flexible in computing the scattering properties of particles with complex morphological configurations.     

The determination of modal damping ratios and natural frequencies from bispectrum modeling

We present a new signal processing and testing technique by using a higher statistical moment, the bispectrum, to determine the damping ratio and natural frequency of offshore structures excited by both unexpected Gaussian forces and known non-Gaussian driving forces. Due to unexpected exciting forces, such as turbulence, in the ocean, environment, the transfer functions of offshore structures are not determined through operating a known driving force and measuring its response. In order to overcome this problem, some of the existing techniques try to model the unexpected forces as white Gaussian forces or almost white Gaussian forces and determine the modal parameters from the response only. Others try to average the input and output to suppress unexpected parts. Our method uses third-order moments to keep the influence of the unexpected Gaussian forces away from the determination of the transfer function of the structure which has linear properties. We model the third-order moment property of the response function with a bispectral model. The modal parameters can be calculated from the estimated model's coefficients. The method has been proven by a number of simulations.     

Risk factors for high blood lead levels among the general population in Taiwan

BACKGROUND: Tacrolimus (FK506) may represent a major advance in the management of allograft rejection after solid organ transplantation. In August 1994 a European heart transplantation pilot study was initiated to assess the efficacy and safety of tacrolimus when administered exclusively through an oral route. METHODS: Eighty-two heart transplant recipients were randomized to treatment (2:1 ratio) with either tacrolimus- (n=54) or cyclosporine-based therapy (n=28). RESULTS: No significant differences were evident between the two treatment groups in either rejection or survival rates at 1 year. Kaplan-Meier estimates of the freedom from rejection were 26.3% and 18.5%, respectively, for the tacrolimus and cyclosporine treatment groups (p=.444). Survival rates were 79.6% and 92.9% (p=.125). At 3 of the 5 centers, patients received antithymocyte globulin during the immediate postoperative period and fared better than those who did not (with acute rejection-free rates of 49.2% and 26.7% for tacrolimus and cyclosporine, respectively [p=.080], as opposed to 7.1% and 8.3% [p=.965]; patient survival rates of 84.6% and 93.3% [p=.382] vs 75.0% and 92.3% [p=.243]). The overall rates of infection, impaired renal function (31.5% vs 21.4%), and glucose intolerance (7.0% vs 4.3%) did not differ significantly between the tacrolimus and cyclosporine treatment groups. Tacrolimus seemed to possess an advantage with regard to a reduced requirement for antihypertensive therapy (59.5% vs 87.5%, p=.025). CONCLUSIONS: Immunosuppression with oral tacrolimus provides a viable alternative to treatment with cyclosporine, particularly when administered in conjunction with antibody therapy. Further studies are warranted to optimize the administration of tacrolimus in this indication.     

Contribution of phospholipase C-beta3 phosphorylation to the rapid attenuation of opioid-activated phosphoinositide response

Our objective was: (1) to determine the appropriate dose of new ultrasmall superparamagnetic iron oxide particles for magnetic resonance angiography (MRA). This agent comprised of a single iron oxide crystal stabilized with a carbohydrate-polyethylene glycol coat (PEG-Ferron/NC 100150 injection); (2) to determine the proper flip angle for PEG-Ferron-enhanced 3 D time-of-flight (TOF) MRA sequence; and (3) to compare the enhancement of peripheral vessels following PEG-Ferron and GdDTPA-BMA. MRA parameters were: TR/TE = 50/2.1 ms, NEX = 1, FOV = 30 x 30 x 1.8 cm, and matrix = 256 x 128 x 64. In anesthetized beagle dogs (n = 10), the effects of PEG-Ferron and GdDTPA-BMA on regional signal were monitored for 45 min and compared. The lowest dose of PEG-Ferron (0.05 mmol/kg) produced the best enhancement of primary, secondary and tertiary vessels. The flip angle 60 degrees provided better enhancement than 20 degrees on contrast enhanced images. Unlike GdDTPA-BMA, PEG-Ferron allowed prolonged delineation (> 45 min) of the pelvis and lower extremities circulation. PEG-Ferron provided greater Contrast-to-noise ratio CNR (80.2 +/- 6.2, P < 0.05) than GdDTPA-BMA (63.5 +/- 2.5). It may be possible for blood pool contrast-enhanced 3 D TOF MRA to provide valuable information for visualization of vascular tree including guiding interventions.     

Regulation of periodontal ligament cell functions by interleukin-1beta

Periodontal ligament (PDL) cells maintain the attachment of the tooth to alveolar bone. These cells reside at a site in which they are challenged frequently by bacterial products and proinflammatory cytokines, such as interleukin-1beta (IL-1beta), during infections. In our initial studies we observed that IL-1beta down-regulates the osteoblast-like characteristics of PDL cells in vitro. Therefore, we examined the functional significance of the loss of the PDL cell's osteoblast-like characteristics during inflammation. In this report we show that, during inflammation, IL-1beta can modulate the phenotypic characteristics of PDL cells to a more functionally significant lipopolysaccharide (LPS)-responsive phenotype. In a healthy periodontium PDL cells exhibit an osteoblast-like phenotype and are unresponsive to gram-negative bacterial LPS. Treatment of PDL cells with IL-1beta inhibits the expression of their osteoblast-like characteristics, as assessed by the failure to express transforming growth factor beta1 (TGF-beta1) and proteins associated with mineralization, such as alkaline phosphatase and osteocalcin. As a consequence of this IL-1beta-induced phenotypic change, PDL cells become responsive to LPS and synthesize proinflammatory cytokines. The IL-1beta-induced phenotypic changes in PDL cells were transient, as removal of IL-1beta from PDL cell cultures resulted in reacquisition of their osteoblast-like characteristics and lack of LPS responsiveness. The IL-1beta-induced phenotypic changes occurred at concentrations that are frequently observed in tissue exudates during periodontal inflammation (0.05 to 5 ng/ml). The results suggest that, during inflammation in vivo, IL-1beta may modulate PDL cell functions, allowing PDL cells to participate directly in the disease process by assuming LPS responsiveness at the expense of their normal structural properties and functions.