A cytosolic activity distinct from crm1 mediates nuclear export of protein kinase inhibitor in permeabilized cells

The leucine-rich nuclear export signal (NES) is used by a variety of proteins to facilitate their delivery from the nucleus to the cytoplasm. One of the best-studied examples, protein kinase inhibitor (PKI), binds to the catalytic subunit of protein kinase A in the nucleus and mediates its rapid export to the cytoplasm. We developed a permeabilized cell assay that reconstitutes nuclear export mediated by PKI, and we used it to characterize the cytosolic factors required for this process. The two-step assay involves an import phase and an export phase, and quantitation is achieved by digital fluorescence microscopy. During the import phase, a fluorescent derivative of streptavidin is imported into the nuclei of digitonin-permeabilized HeLa cells. During the export phase, biotinylated PKI diffuses into the nucleus, binds to fluorescent streptavidin, and mediates export of the complex to the cytoplasm. Nuclear export of the PKI complex is cytosol dependent and can be stimulated by addition of the purified NES receptor, Crm1. HeLa cell cytosol treated with N-ethylmaleimide (NEM) or phenyl-Sepharose to inactivate or deplete Crm1, respectively, is still fully active in the PKI export assay. Significantly, the export activity can be depleted from cytosol by preadsorption with a protein conjugate that contains a functional NES. These data indicate that cytosol contains an export activity that is distinct from Crm1 and is likely to correspond to an NES receptor.     

增加折叠电流保护的电路

对许多需要数安培供电电流的应用而言，三端输出可调线性稳压器有易用．低成本和完全片上过载保护等特点，如美国国家半导体公司的LM317。增加数个元器件可以使三端稳压器具备一种高速度的短路限流能力，从而提高可靠性。限流器可将最大输出电流限制在一个安全的恒定水平IMAX上，避免稳压器的损坏。当出现一个故障状况时，传输晶体管上的功耗近似于VIN×IMAX。要让设计的稳压器能承受过载，就需要谨慎地选择元器件的额定值（经常是留了过大的余地），除非可以在故障发生时降低或折叠（foldback）输出电流。     

Asymptotic Bias and Variance of Conventional Bispectrum Estimates for 2-D Signals

In this paper, we derive the asymptotic bias and variance of conventional bispectrum estimates of 2-D signals. Two methods have been selected for the estimation: the first one – the indirect method – is the Fourier Transform of the weighted third order moment, while the second one – the direct method – is the expectation of the Fourier component product. Most of the developments are known for 1-D signals and the first contribution of this paper is the rigorous extension of the results to 2-D signals. The calculation of the bias of the direct method is a totally original contribution. Nevertheless, we did all calculations (bias and variance) for both method in order to be able to compare the results. The second contribution of this paper consists of the comparison of the theoretical bispectrum estimate bias and variance with the measured bias and variance for two 2-D signals. The first studied signal is the output of a non-minimal phase linear system driven by a non-symmetric noise. The second signal is the output of a non-linear system with Gaussian input data. In order to assess the results, we performed the comparison for both methods with different sets of parameters. We show that the maximum bias coefficient is the one of the 1-D case multiplied by the dimensionality of the signal for both methods. We also show that the estimate variance coefficient is the 1-D case coefficient with a power equal to the signal dimensionality.Received October 21, 2002; Revised December 2003; Accepted March 25, 2004; First Online Version published in December 2004     

Accelerated study on lysozyme deposition on poly(HEMA) contact lenses

A technique was developed to accelerate lysozyme deposition on poly(HEMA) contact lenses and measure the amounts of the deposited lysozyme. This technique was for evaluation of bendazac lysine solution, a contact lens cleaning and wetting solution. Effect of temperature on lysozyme deposition on poly(HEMA) contact lenses was examined. Five temperatures ranging from 25 degrees C to 90 degrees C were chosen to examine the temperature effect. The amounts of lysozyme deposited on poly(HEMA) contact lenses at 25 C and 60 C were 0.27 microg/lens and 0.61 microg/lens, respectively. The amount increased sharply to 23 microg/lens at 70 degrees C with the maximum of 31 microg/lens at 90 degrees C. Kinetics of lysozyme deposition on poly(HEMA) contact lenses was examined at 80 degrees C. Lysozyme deposition increased sharply during the first 2 h and reached a plateau after 2 h. Effectiveness of various cleaning procedures was examined using bendazac lysine solution. When the contact lenses were washed without rubbing with fingers, the bendazac lysine reduced the amount of deposited lysozyme by more than 40% from 18.3 microg/lens to 10.6 microg/lens. The effect of bendazac lysine was most prominent when the contact lenses were shaken during storage in the presence of lysozyme in solution. If the contact lenses were cleaned by rubbing with fingers, the effect of bendazac lysine solution on the prevention of lysozyme deposition was negligible.     

Subdiaphragmatic and intrathoracic paraspinal malignant peripheral nerve sheath tumors: a clinicopathologic study of 25 patients and 26 tumors

BACKGROUND: To determine the effects of anatomic site on the presentation and diagnosis of malignant peripheral nerve sheath tumors (MPNSTs) and on the treatment and outcomes of the patients, the authors initiated a study of these tumors at different sites. An earlier report described MPNSTs of the buttock and lower extremity, and the current series analyzes those presenting at intrathoracic (IT) and subdiaphragmatic (SD) paraspinal sites. METHODS: The authors reviewed data on patients with paraspinal MPNSTs who were seen at Memorial Hospital during the period 1960-1995 and for whom histologic slides were available. Various clinicopathologic parameters and their effects on patient outcomes were examined. RESULTS: Twenty-five patients with 26 tumors were evaluated. Seven tumors were IT and 19 were SD; 60% of the patients had neurofibromatosis type 1 (NF1). Most patients presented with pain, and a diagnostic delay (of 3 months to 2 years) was often noted. Mean tumor sizes for SD and IT tumors were 14.3 cm and 6.6 cm, respectively. Most MPNSTs were composed of spindle cells in fascicles. Twenty-seven percent exhibited divergent differentiation. Twenty-four tumors were high grade, and a low grade component was identified in 8 tumors. Surgical resection was attempted for 23 tumors (88%), but complete resection was achieved in only 6 cases (23%). Eighty percent of the patients died of their tumors, 2-year and 5-year survival rates were 35% and 16%, and median survival was 8.5 months. Significant prognostic factors were tumor size <5 cm, the presence of a low grade component, and complete tumor resection. CONCLUSIONS: Paraspinal MPNSTs have more aggressive behavior than peripherally located tumors, mainly because of the difficulty encountered in resecting them completely. Prognoses of patients with MPNST at this site appear to be affected by resection status, tumor size, and tumor grade.     

Response of two populations of holm oak (Quercus rotundifolia Lam.) to sulfur dioxide

Experiments were carried out with seedlings of Quercus rotundifolia Lam., an evergreen schlerophyllous tree typical of the Spanish Mediterranean climate environments. Fruits were collected in two distant (800 km) populations located in the center (southern Spain) and northern border (northern Spain) of the area of distribution of the species. One-month-old potted plants were grown for 130 days in an enriched atmosphere of SO2 (0.23 ppm, 14 h/day) in controlled (growth chamber) conditions. Both northern and southern plants underwent a significant decrease in growth rate as a consequence of the treatment. Even so, plants appear to be quite resistant to SO2 compared with either more temperate or more productive species. The southern population was more sensitive to the treatment, as reflected by the bigger decrease in both growth and photosynthetic rates. Differences in resistance appear to be related to the biogeographic origin of the populations studied, which underlines the importance of biogeographic aspects in studies of resistance to air pollutants.     

Environmental health implications of heavy metal pollution from car tires

OBJECTIVE: To determine the effect of retinoic acid on wound healing of laser burns to explants of porcine retinal pigment epithelium (RPE). DESIGN: With the help of a mirrored attachment to a Coherent argon laser, laser burns (spot size 100 mu, power 500 mW, duration 0.2 seconds) were performed in 12 explants. The explants were then cultured in the presence (six explants) or absence (six explants) of 1 microM retinoic acid. Two explants in either group were recovered 4, 8 and 12 days after treatment for histologic study and scanning electron microscopy. OUTCOME MEASURE: Mean burn area remaining after culture. RESULTS: The area of wound remaining unhealed in the presence of retinoic acid was significantly larger than the area of wound remaining unhealed in the absence of retinoic acid at each time point. CONCLUSIONS: Retinoic acid inhibited wound healing of laser burns to porcine RPE. This effect may illustrate a role for retinoic acid in the pharmacotherapy of proliferative disorders.