A novel serine protease inhibitor from Bungarus fasciatus venom

By Sephadex G-50 gel filtration, cation-exchange CM-Sephadex C-25 chromatography and reversed phase high-performance liquid chromatography (HPLC), a novel serine protease inhibitor named bungaruskunin was purified and characterized from venom of Bungarus fasciatus. Its cDNA was also cloned from the cDNA library of B. fasciatus venomous glands. The predicted precursor is composed of 83 amino acid (aa) residues including a 24-aa signal peptide and a 59-aa mature bungaruskunin. Bungaruskunin showed maximal similarity (64%) with the predicted serine protease inhibitor blackelin deduced from the cDNA sequence of the red-bellied black snake Pseudechis porphyriacus. Bungaruskunin is a Kunitz protease inhibitor with a conserved Kunitz domain and could exert inhibitory activity against trypsin, chymotrypsin, and elastase. By screening the cDNA library, two new B chains of beta-bungarotoxin are also identified. The overall structures of bungaruskunin and beta -bungarotoxin B chains are similar; especially they have highly conserved signal peptide sequences. These findings strongly suggest that snake Kunitz/BPTI protease inhibitors and neurotoxic homologs may have originated from a common ancestor. (c) 2007 Elsevier Inc. All rights reserved.

Properties of the proton-evoked currents and their modulation by Ca2+ and Zn2+ in the acutely dissociated hippocampus CA1 neurons

The characterization of acid-sensing ion channel (ASIC)-like currents has been reported in hippocampal neurons in primary culture. However, it is suggested that the profile of expression of ASICs changes in culture. In this study, we investigated the properties of proton-activated current and its modulation by extracellular Ca2+ and Zn2+ in neurons acutely dissociated from the rat hippocampal CA1 using conventional whole-cell patch-clamp recording. A rapidly decaying inward current and membrane depolarization was induced by exogenous application of acidic solution. The current was sensitive to the extracellular proton with a response threshold of pH 7.0-6.8 and the pH(50) Of 6.1, the reversal potential close to the Na+ equilibrium potential. It had a characteristic of acid-sensing ion channels (ASICs) as demonstrated by its sensitivity to amiloride (IC50 = 19.6 +/- 2.1 muM). Either low [Ca2+](0) or high [Zn2+](0) increased the amplitude of the current. All these characteristics are consistent with a current mediated through a mixture of homomeric ASIC1a and heteromeric ASIC1a + 2a channels and closely replicate many of the characteristics that have been previously reported for hippocampal neurons cultured for a week or more, indicating that culture artifacts do not necessarily flaw the properties of ASICs. Interestingly, we found that high [Zn2+] (>10(-4) M) slowed the decay time constant of the ASIC-like current significantly in both acutely dissociated and cultured hippocampal neurons. In addition, the facilitating effects of low [Ca2+](0) and high [Zn2+](0) on the ASIC-like current were not additive. Since tissue acidosis, extracellular Zn elevation and/or Ca2+ reduction occur concurrently under some physiological and/or pathological conditions, the present observations suggest that hippocampal ASICs may offer a novel pharmacological target for therapeutic invention. (C) 2004 Elsevier B.V. All rights reserved.

HORMONAL-REGULATION OF TISSUE-TYPE PLASMINOGEN-ACTIVATOR AND PLASMINOGEN-ACTIVATOR INHIBITOR TYPE-1 IN CULTURED MONKEY SERTOLI CELLS

Sertoli cells play a central role in the control and maintenance of spermatogenesis. Isolated Sertoli cells of mouse and rat testes have been shown to secrete plasminogen activator (PA) and a plasminogen activator inhibitor type-1 (PAI-1) in culture. In this study, we have investigated the hormonal regulation of PA and PAI-1 activities in cultured monkey Sertoli cells. Sertoli cells (5x10(5) cells/well) isolated from infant rhesus monkey testes were preincubated at 35 degrees C for 16 h in 24-well plates precoated with poly(D-lysine) (5 mu g/cm(2)) in 0.5 mi McCoy's 5a medium containing 5% of fetal calf serum and further incubated for 48 h in 0.5 mi serum-free medium with or without various hormones or other compounds, PA as well as PAI-1 activities in the conditioned media were assayed by fibrin overlay and reverse fibrin autography techniques respectively. The Sertoli cells in vitro secreted only tissue-type PA (tPA), no detectable amount of urokinase-type PA (uPA) could be observed, Monkey Sertoli cells were also capable of secreting PAI-1, Immunocytochemical studies indicated that both tPA and PAI-1 positive staining localized in the Sertoli cells, spermatids and residual bodies of the seminiferous epithelium; Northern blot analysis further confirmed the presence of both tPA and PAI-1 mRNA in monkey Sertoli cells. Addition of follicle-stimulating hormone (FSH) or cyclic adenosine monophosphate (cAMP) derivatives or cAMP-generating agents and gonadotrophin-releasing hormone (GnRH) agonist or phorbol ester (PMA) to the cell culture significantly increased tPA activity. PAI-1 activity in the culture was also enhanced by these reagents except 8-bromo-dibutyryl-cAMP, forskolin and 3-isobutyl-1-methylxanthin (MIX) which greatly stimulated tPA activity, whereas decreased PAI-1 activity, implying that neutralization of PAI-1 activity by tile high level of tPA in the conditioned media may occur. These data suggest that increased intracellular signals which activate protein kinase A (PKA), or protein kinase C (PKC) can modulate Sertoli cell tPA and PAI-1 activities, The concomitant induction of PA and PAI-1 by the same reagents in the Sertoli cells may reflect a finely tuned regulatory mechanism in which PAI-1 could limit the excession of the proteolysis.

Purification, characterization and primary structure of a chymotrypsin inhibitor from Naja atra venom

A chymotrypsin inhibitor, designated NA-CI, was isolated from the venom of the Chinese cobra Naja atra by three-step chromatography. It inhibited bovine (x-chymotrypsin with a K-i of 25 nM. The molecular mass of NA-CI was determined to be 6403.8 Da by matrix-assisted laser-desorption ionization time-of-flight (MALDI-TOF) analysis. The complete amino acid sequence was determined after digestion of S-carboxymethylated inhibitor with Staphylococcus aureus V8 protease and porcine trypsin. NA-CI was a single polypeptide chain composed of 57 amino acid residues. The main contact site with the protease (PI) has a Phe, showing the specificity of the inhibitor. NA-CI shared great similarity with the chymotrypsin inhibitor from Naja naja venom (identities = 89.5%) and other snake venom protease inhibitors. (C) 2003 Elsevier Inc. All rights reserved.

Optimization of thermal material in a flux pump system with high temperature superconductor

Superconductors are known for the ability to trap magnetic field. A thermally actuated magnetization (TAM) flux pump is a system that utilizes the thermal material to generate multiple small magnetic pulses resulting in a high magnetization accumulated in the superconductor. Ferrites are a good thermal material candidate for the future TAM flux pumps because the relative permeability of ferrite changes significantly with temperature, particularly around the Curie temperature. Several soft ferrites have been specially synthesized to reduce the cost and improve the efficiency of the TAM flux pump. Various ferrite compositions have been tested under a temperature variation ranging from 77K to 300K. The experimental results of the synthesized soft ferrites-Cu 0.3 Zn 0.7Ti 0.04Fe 1.96O 4, including the Curie temperature, magnetic relative permeability and the volume magnetization (emu/cm3), are presented in this paper. The results are compared with original thermal material, gadolinium, used in the TAM flux pump system.-Cu 0.3 Zn 0.7Ti 0.04 Fe 1.96O 4 holds superior characteristics and is believed to be a suitable material for next generation TAM flux pump. © 2011 IEEE.

UV photoconductivity characteristics of ZnO nanowire field effect transistor treated by proton irradiation

We investigated the UV photoconductivity characteristics of ZnO nanowire field effect transistors (FETs) irradiated by proton beams. After proton beam irradiation (using a beam energy of 10 MeV and a fluence of 10 12 cm -2), the drain current and carrier density in the ZnO nanowire FETs decreased, and the threshold voltage shifted to the positive gate bias direction due to the creation of interface traps at the SiO 2/ZnO nanowire interface by the proton beam. The interface traps produced a higher surface barrier potential and a larger depletion region at the ZnO nanowire surface, affecting the photoconductivity and its decay time. The UV photoconductivity of the proton-irradiated ZnO nanowire FETs was higher and more prolonged than that of the pristine ZnO nanowire FETs. The results extend our understanding of the UV photoconductivity characteristics of ZnO nanowire devices and other materials when irradiated with highly energetic particles. © 2012 Elsevier B.V. All rights reserved.

Risk based lifetime costs assessment of a ground source heat pump (GSHP) system design: Methodology and case study

Space heating accounts for a large portion of the world's carbon dioxide emissions. Ground Source Heat Pumps (GSHPs) are a technology which can reduce carbon emissions from heating and cooling. GSHP system performance is however highly sensitive to deviation from design values of the actual annual energy extraction/rejection rates from/to the ground. In order to prevent failure and/or performance deterioration of GSHP systems it is possible to incorporate a safety factor in the design of the GSHP by over-sizing the ground heat exchanger (GHE). A methodology to evaluate the financial risk involved in over-sizing the GHE is proposed is this paper. A probability based approach is used to evaluate the economic feasibility of a hypothetical full-size GSHP system as compared to four alternative Heating Ventilation and Air Conditioning (HVAC) system configurations. The model of the GSHP system is developed in the TRNSYS energy simulation platform and calibrated with data from an actual hybrid GSHP system installed in the Department of Earth Science, University of Oxford, UK. Results of the analysis show that potential savings from a full-size GSHP system largely depend on projected HVAC system efficiencies and gas and electricity prices. Results of the risk analysis also suggest that a full-size GSHP with auxiliary back up is potentially the most economical system configuration. © 2012 Elsevier Ltd.

Design consideration of a circular type magnetic flux pump device

As a variation of the thermally actuated flux pump and the linear type magnetic flux pump (LTMFP), the circular type magnetic flux pump (CTMFP) device is proposed to magnetize a circular shape type-II superconducting thin film and bulk. The basic concept is the same as the thermally actuated flux pump: a circularly symmetric traveling magnetic field is generated below a circular shape superconductor to increase its trapping field. However, this traveling field is created by the three phase windings instead of heating gadolinium block. Apart from the LTMFP, the three phase windings are wound concentrically instead of linearly. The speed of the traveling field is controlled by the AC frequency and the magnitude of the field is controlled by the magnitudes of AC currents. In addition, a coil with DC current is wound around the three phase windings to provide a background field. The concept design is presented in this paper. The magnetic waveforms are analysed numerically by the COMSOL 3.5a software. The impedances of the three phase windings are calculated and a corresponding circuit design is presented. This rig can be used as an advanced tool to study the flux pump behavior of a circular shape superconductor. © 2002-2011 IEEE.

A finite element model of magnetization of superconducting bulks using a solid-state flux pump

Superconductors have a bright future; they are able to carry very high current densities, switch rapidly in electronic circuits, detect extremely small perturbations in magnetic fields, and sustain very high magnetic fields. Of most interest to large-scale electrical engineering applications are the ability to carry large currents and to provide large magnetic fields. There are many projects that use the first property, and these have concentrated on power generation, transmission, and utilization; however, there are relatively few, which are currently exploiting the ability to sustain high magnetic fields. The main reason for this is that high field wound magnets can and have been made from both BSCCO and YBCO, but currently, their cost is much higher than the alternative provided by low-Tc materials such as Nb3Sn and NbTi. An alternative form of the material is the bulk form, which can be magnetized to high fields. This paper explains the mechanism, which allows superconductors to be magnetized without the need for high field magnets to perform magnetization. A finite-element model is presented, which is based on the E-J current law. Results from this model show how magnetization of the superconductor builds up cycle upon cycle when a traveling magnetic wave is induced above the superconductor. © 2011 IEEE.