The relationship between electropermeabilization and cell cycle and cell size of Saccharomyces cerevisiae

Yeast cells (Sacchromyces cerevisiae) in 0.9% NaCl solution containing phloxine B (dye) were treated by an application of a rectangular electric pulse. We input microscopic images of the yeast suspensions after the application into a computer, and measured whether each cell dyes or not, the phase in the cell cycle, and each cell size, using the software we had developed. After those measurements, we discussed the relationship between the yield of electropermeabilization (the ratio of dyed cells to the total cell number) and the phase in the cell cycle, and cell size. From the results, it was found that the yeast cells from S-phase to M-phase (S-M phase) in the cell cycle tend to be more permeated than G1-phase yeast cells, and in both phases the yield decreases with the increase in cell size.     

MgATP-induced conformational changes in a single myosin molecule observed by atomic force microscopy: periodicity of substructures in myosin rods

This paper discusses the conformational changes in a single myosin molecule directly observed using atomic force microscopy (AFM). The myosin molecules were pretreated in rigor solutions without MgATP or in relaxed solutions with various concentrations of MgATP. The images of these molecules were obtained using a tapping mode AFM. The results indicate that the orientation of the myosin's heads and tail strongly depend on the MgATP concentration. Without using MgATP, almost all of the myosin molecules are in the extended form; however, when MgATP is used, the molecules bend according to the level of MgATP concentration. The mean-square end-to-end distance of the myosin molecules is significantly shorter with p[MgATP] = 4 than with p[MgATP] = 6. The rod region did not show the same level of intensity along their length in the extended form. The rods exhibited clusters of discontinuity, which were identified as substructures. The size of these substructures change at intervals that are multiples of 14.3-14.5 nm, which reflects the periodicity of the alpha-helical coiled coils. The substructure clusters also correspond to the myosin crossbridge spacing in muscles (14.3 or 43 nm). These results suggest that the myosin's head bends in conjunction with the bending or tilting in the helical substructures. Conformational changes of the myosin molecule induced by MgATP seem to mimic the molecular motions in a muscle's force generation process.     

A novel anionic surfactant templating route for synthesizing mesoporous silica with unique structure

Anionic surfactants are used in greater volume than any other surfactants because of their highly potent detergency and low cost of manufacture. However, they have not been used as templates for synthesizing mesoporous silica. Here we show a templating route for preparing mesoporous silicas based on self-assembly of anionic surfactants and inorganic precursors. We use aminosilane or quaternized aminosilane as co-structure-directing agent (CSDA), which is different from previous pathways. The alkoxysilane site of CSDA is co-condensed with inorganic precursors; the ammonium site of CSDA, attached to silicon atoms incorporated into the wall, electrostatically interacts with the anionic surfactants to produce well-ordered anionic-surfactant-templated mesoporous silicas (AMS). These have new structures with periodic modulations as well as two-dimensional hexagonal and lamellar phases. The periodic modulations may be caused by the coexistence of micelles that differ in size or curvature, possibly owing to local chirality. These mesoporous silicas provide a new family of mesoporous materials as well as shedding light on the structural behaviour of anionic surfactants.     

Superfluid-Normal Interface of 4He Near a Wall

We observed a profile of nonequilibrium superfluid-normal (SN) interface of ⁴He near a vertical wall. A glass, brass or copper wall was used. The SN interface was produced by cooling liquid ⁴He in a bath from the bottom, where liquid ⁴He was pumped through a flow impedance in order to cool down the liquid. Superfluid (Normal fluid) occupied the lower (upper) part of the bath. The SN interface was visualized by three methods: simple visualization, shadowgraphy and schlieren method. The interface touched a vertical glass wall at almost 90°. A large hollow was observed near a brass wall which had intermediate thermal conductivity. Downward flow was observed on a copper wall due to the very good thermal conductivity of the wall. Various types of interface profile were observed depending on the thermal conductivity of the wall used.      

A multi-spectral non-local method for retrieval of boundary layer cloud properties from optical remote sensing data

A multi-spectral non-local (MSN) method is developed for advanced retrieval of boundary layer cloud properties from remote sensing data, as an alternative to the independent pixel approximation (IPA) method. The non-local method uses data at both the target pixel and neighboring pixels to retrieve cloud properties such as pixel-averaged cloud optical thickness and effective droplet radius. Radiance data to be observed from space were simulated by a three-dimensional (3D) radiation model and a stochastic boundary layer cloud model with two-dimensional (horizontal and vertical) variability in cloud liquid water and effective radius. An adiabatic assumption is used for each cloud column to model the geometrical thickness and vertical profiles of cloud liquid water content and effective droplet radius, neglecting drizzle and cloud brokenness for simplicity. The dependence of radiative smoothing and roughening on horizontal scale, optical thickness and single scattering albedo are investigated. Then, retrieval methods using 250-m horizontal resolution data onboard new generation satellites are discussed. The regression model for the MSN method was trained based on datasets from numerical simulations. The training was performed with respect to various domain averages of optical thickness and effective radius, because smoothing and roughening effects are strongly dependent on the two variables. Retrieval accuracy is discussed here with datasets independent of those used in the training, towards assessing the generality of the technique. It is demonstrated that retrieval accuracy of cloud optical thickness, which is often retrieved from single-spectral visible-wavelength data, is improved the most using neighboring pixel data and secondly using multi-spectral data, and ideally with both. When the IPA retrieval method is applied to optical thickness and effective radius, the root-mean-square relative errors can be 15-90%, depending on solar and view directions. In contrast, the MSN method has errors of 4-10%, which is smaller than IPA by a factor of 2-10. It is also suggested that the accuracy of the MSN method is insensitive to some assumptions in the inhomogeneous cloud input data used to train the regression model.     

Ion-beam processing of single crystal diamond using SOG mask

Various ion-beam etching characteristics of diamond and selectivity between diamond and spin-on-glass (SOG) were examined. The maximum selectivity of diamond and SOG was 12.7 in oxygen reactive ion-beam etching process at 100 V acceleration voltage. Using this etching condition and dot-shaped SOG mask, conical diamond field electron emitter arrays with 30 nm curvature radius, 2.58 μm base radius and 5.86 μm height were fabricated.     

Angiographic evaluation of culprit lesions in acute coronary syndrome: relation to the original site on previous coronary angiography

Recent studies suggest that release of cytokines during inflammatory states such as septic shock leads to hypocholesterolemia. To examine whether tumor necrosis factor alpha (TNF), which is the major cytokine in inflammatory disease, causes hypocholesterolemia, we measured serum levels of total (bioactive and receptor-bound) TNF, cholesterol, Apo B, and Apo A1 in seven patients with septic shock over a period of 8 days. Since elevated serum TNF levels are accompanied by the release of soluble TNF receptors, levels of TNF receptors p55 and p75 were also measured. Patients with septic shock had significantly higher serum TNF and TNF receptor levels compared with healthy controls. Increased cytokine levels were accompanied by a significant decline in total serum cholesterol apolipoprotein A1 and B. In vitro studies with cultured human skin fibroblasts, human umbilical vein endothelial cells, and HepG2 hepatoma cells showed that TNF increased the degradation of 125I-labeled low-density lipoprotein in all the cell lines tested. Recombinant soluble TNF receptors inhibited the TNF-induced stimulation of low-density lipoprotein receptor in a concentration-dependent manner. However, the calculated ratio of TNF receptors to total TNF measured in serum of these patients was not able to counteract the stimulatory effect of TNF, possibly due to the higher molar excess of TNF receptors required to achieve this effect in vitro. Our data strengthen the hypothesis that serum values of total TNF determine the extent of hypocholesterolemia during sepsis and septic shock despite the presence of a high concentration of TNF receptors. Studies with recombinant TNF also confirm the role of TNF in hypocholesterolemia in inflammation.     

Effects of Al3+ and Be2+ ions combined with NaF on ciliary process adenylyl cyclase activity and aqueous humor dynamics in the rabbit eye

PURPOSE: The activity of Al3+, Ga3+, and Be2+ ions in the presence of NaF to directly activate G-proteins was investigated by their potentiative effect on forskolin (FSK)-activated adenylyl cyclase in rabbit ciliary process membranes and their effects on aqueous humor dynamics in vivo. METHODS: Adenylyl cyclase (AC) was determined by radiometric conversion of ATP to cAMP by the particulate fraction of rabbit ciliary processes. Intravitreal injections of sterile solutions of analytical grade salts were made into the center of the vitreous in a volume of 20 microliters. Intraocular pressure, aqueous humor flow, and uveoscleral outflow measurements were made by pneumatonometry, fluorophotometry, and fluorescein-dextran method, respectively. Outflow facility was determined by tonography in the intact eyes and by two-level constant pressure perfusion in cannulated eyes. RESULTS: Both Al3+ (EC50, 40 mumol/l) and Be2+ (EC50, 11 mumol/l) in the presence of 0.5-2 mM NaF activated the stimulatory G-protein Gs. Ga3+ was ineffective and did not antagonize the activation by Al3+. Intravitreal injections of Al3+ (1 mumol/eye) or Be2+ (0.5 or 1 mumol/eye) had no significant intraocular pressure (IOP) effect, nor did 1.5 or 3 mumol/eye of NaF, but when either cation was injected together with NaF, IOP decreased by up to 40% for up to 140 hr. At the time of maximum IOP effect (72 hr) aqueous humor flow determined by fluorophotometry was decreased in BeCl2+ NaF-treated eyes by 40% relative to BeCl2-treated eyes; however, tonographic facility of outflow was unaffected. Uveoscleral flow was also decreased by 38% in BeCl2+ NaF treated eyes. CONCLUSIONS: These findings support the hypothesis that Gs activation of ciliary process adenylyl cyclase decreases aqueous humor formation rate in rabbit eyes, and that activation of G-proteins mediates contraction of ciliary muscles causing a decrease of aqueous humor outflow via the uveoscleral route. The results suggest that G-proteins putatively involved in trabecular facility changes are less sensitive to activation by BeF3- than are other parameters of aqueous humor dynamics.     

Models for circular data from time series spectra

Circular data are those for which the natural support is the unit circle and its toroidal extensions. Numerous constructions have been proposed which can be used to generate models for such data. We propose a new, very general, one based on the normalization of the spectra of complex-valued stationary processes. As illustrations of the new construction's application, we study models for univariate circular data obtained from the spectra of autoregressive moving average models and relate them to existing models in the literature. We also propose and investigate multivariate circular models obtained from the high-order spectra of stationary stochastic processes generated using linear filtering with an autoregressive moving average response function. A new family of distributions for a Markov process on the circle is also introduced. Results for asymptotically optimal inference for dependent observations on the circle are presented which provide a new paradigm for inference with circular models. The application of one of the new families of spectra-generated models is illustrated in an analysis of wind direction data.     

Statistical analysis for multiplicatively modulated nonlinear autoregressive model and its applications to electrophysiological signal analysis in humans

Modulating the dynamics of a nonlinear autoregressive model with a radial basis function (RBF) of exogenous variables is known to reduce the prediction error. Here, RBF is a function that decays to zero exponentially if the deviation between the exogenous variables and a center location becomes large. This paper introduces a class of RBF-based multiplicatively modulated nonlinear autoregressive (mmNAR) models. First, we establish the local asymptotic normality (LAN) for vector conditional heteroscedastic autoregressive nonlinear (CHARN) models, which include the mmNAR and many other well-known time-series models as special cases. Asymptotic optimality for estimation and testing is described in terms of LAN properties. The mmNAR model indicates goodness-of-fit for surface electromyograms (EMG) using electrocorticograms (ECoG) as the exogenous variables. Concretely, it is found that the negative potential of the motor cortex forces change in the frequency of EMG, which is reasonable from a physiological point of view. The proposed mmNAR model fitting is both useful and efficient as a signal-processing technique for extracting information on the action potential, which is associated with the postsynaptic potential.