Aflatoxin-selective molecularly-imprinted polymer membranes based on acrylate-polyurethane semi-interpenetrating polymer networks

Synthetic analogs of biological receptors able to group-selective recognition of aflatoxins were obtained using the combination of the technique of molecular imprinting with the method of computer modeling. The synthetic receptors were obtained in a form of thin and porous membranes based on semi-interpenetrating polymer networks. The selection of functional monomers able to noncovalent interactions with aflatoxins B1, B2, and G2 was based on the data of computer modeling. Allylamine, diethylaminoethylmethacrylate, and N,N'-methylenebisacrylamide, providing high binding energies with aflatoxins B1, B2, and G2 were selected as functional monomers for the formation of aflatoxin B1-imprinted polymer membranes. It was shown that aflatoxin-B1-imprinted polymeric membranes synthesized using N,N'-methylenebisacrylamide as a functional monomer were characterized with good physico-mechanical properties as well as good adsorbtion capability towards aflatoxin B1. Neglidgible levels of aflatoxin B1 adsorbtion on the surface of blank membranes were observed. High adsorbtion capability of the MIP membranes towards mycotoxins affiliated to the group of aflatoxins was demonstrated, while negligible adsorbtion of ochratoxin A was observed. Therefore, synthetic analogs of biological receptors able to group-selective recognition of aflatoxins in the range 1-1000 ppb were developed.     

Environmentally induced changes in mitochondria and endoplasmic reticulum of Saccharomyces carlsbergensis yeast

The effects of culture environment on the volume density and surface density of mitochondria and endoplasmic reticulum in a facultative yeast were studied. When compared with cells grown aerobically on a nonrepressive substrate, cells grown in the absence of oxygen showed a sharp reduction in both volume density of mitochondria and surface density of the inner mitochondrial membrane (imm) in the remaining mitochondrial profiles. Use of fermentable (repressive) substrates under aerobic conditions restricted the volume density of mitochondria to a much greater extent than the surface density of imm. The range of mitochondrial volume densities in these experiments was 4-11%. Surface density of endoplasmic reticulum (ER) was sensitive to growth rate and in particular to changes in oxygen tension, showing large fluctuations during both anaerobic and aerobic adaptation. These fluctuations in ER are discussed in relation to the known role of this organelle in lipid metabolism.     

Sensor system based on molecular-imprinted polymer membranes for the selective recognition of aflatoxin B1

lymer membranes, synthesized using acrylamide as a functional monomer, were characterized by sufficient mechanical stability and high adsorbtion capability towards aflatoxin B1. The molecularly-imprinted polymer membranes were characterized by the pronounced imprinting effect as well as by insignificant adsorbtion of aflatoxins B2 and G2. Therefore, the synthetic analogues of bioreceptors able to individual recognition of aflatoxin B1 were obtained and used as a basis for the optical sensor system for aflatoxin B1 detection in a concentration range 1-500 ng/ml.     

Water uptake profile response of corn to soil moisture depletion

The effects of soil moisture distribution on water uptake of drip‐irrigated corn were investigated by simultaneously monitoring the diurnal evolution of sap flow rate in stems, of leaf water potential, and of soil moisture, during intervals between successive irrigations. The results invalidate the steady‐state resistive flow model for the continuum. High hydraulic capacitance of wet soil and low hydraulic conductivity of dry soil surrounding the roots damped significantly diurnal fluctuations of water flow from bulk soil to root surface. By contrast, sap flow responded directly to the large diurnal variation of leaf water potential. In wet soil, the relation between the diurnal courses of uptake rates and leaf water potential was linear. Water potential at the root surface remained nearly constant and uniformly distributed. The slope of the lines allowed calculating the resistance of the hydraulic path in the plant. Resistances increased in inverse relation with root length density. Soil desiccation induced a diurnal variation of water potential at the root surface, the minimum occurring in the late afternoon. The increase of root surface water potential with depth was directly linked to the soil desiccation profile. The development of a water potential gradient at the root surface implies the presence of a significant axial resistance in the root hydraulic path that explains why the desiccation of the soil upper layer induces an absolute increase of water uptake rates from the deeper wet layers.     

Plasma density fluctuations measurements by heavy ion beam probe with a multiple cell array detector on the tokamak ISTTOK

It is recognized that the broad-band fluctuations observed in plasmas of thermonuclear devices are the cause of anomalously large energy and particle transport. Strong progress has been achieved on different plasma devices using a heavy ion beam probe (HIBP) diagnostic measuring the plasma potential and density fluctuations. Better understanding of turbulence properties can be obtained with a multiple cell array detector (MCAD) as plasma density fluctuations are measured simultaneously at a number of sample volumes. In this paper, the capabilities of the HIBP with MCAD for plasma density fluctuations measurements on the tokamak ISTTOK are analyzed (including restrictions due to sample volume size and path effects) and compared with preliminary experimental data. The upgrade of ISTTOK HIBP for better diagnostic performance is also discussed.     

Fluctuations in membrane current driven by intracellular calcium in cardiac Purkinje fibers

Spontaneous oscillatory fluctuations in membrane potential are often observed in heart cells, but their basis remains controversial. Such activity is enhanced in cardiac Purkinje fibers by exposure to digitalis or K-free solutions. Under these conditions, we find that voltage noise is generated by current fluctuations that persist when membrane potential is voltage clamped. Power spectra of current signals are not made up of single time-constant components, as expected from gating of independent channels, but are dominated by resonant characteristics between 0.5 and 2 HZ. Our evidence suggests that the periodicity arises from oscillatory variations in intracellular free Ca that control ion movements across the surface membrane. The current fluctuations are strongly cross-correlated with oscillatory fluctuations in contractile force, and are inhibited by removing extracellular Ca or exposure to D600. Chelating intracellular Ca with injected EGTA also abolishes the current fluctuations. The oscillatory mechanism may involve cycles of Ca (or Sr) movement between sarcoplasmic reticulum and myoplasm, as previously suggested for skinned cardiac preparations. Our experiments in intact cells indicate that changes in surface membrane potential can modulate cytoplasmic Ca oscillations in frequency and perhaps amplitude as well. A two-way interaction between surface membrane potential and intracellular Ca stores may be a common feature of heart, neuron, and other cell types.     

Displacement of the electron cyclotron resonance heating region and time evolution of the characteristics of short-wavelength turbulence in the 3D magnetic configuration of the L-2M stellarator

Reflection of the heating extraordinary microwave incident obliquely onto the surface of the electron cyclotron resonance (ECR) at the second harmonic of the electron gyrofrequency in the 3D magnetic configuration of the L-2M stellarator was studied experimentally. The plasma was heated using two gyrotrons with a total power of 600–700 kW, the specific heating power being 2.4–2.8 MW/m³. The displacement of the ECR region in the course of heating was monitored by measuring the phase of the reflected extraordinary wave. It is found that the growth of the plasma density is accompanied by the displacement of the ECR heating region from the center of the plasma column toward its periphery. The coefficient of reflection of the heating microwave beam from the ECR region was measured. The spectra of short-wavelength (k _s ≈ 30 cm^?1) plasma density fluctuations were explored by analyzing backscattered microwave radiation. A tenfold increase in the energy of short-wavelength density fluctuations and the growth of the spectral density of fluctuations in the frequency range of 0.3–1.5 MHz were observed.     

Effect of Urey-Bradley-Shimanouchi force field on the harmonic dynamics of proteins.

A normal mode analysis of bovine pancreatic trypsin inhibitor is carried out by using a Urey-Bradley-Shimanouchi potential energy function. The density of vibrational states, the magnitudes, and time scales of the atomic fluctuations are compared with experimental and theoretical results obtained by the more commonly used potential energy functions. The atomic fluctuations of Lys-15 are subject to extensive considerations as this residue is buried in the trypsin specificity pocket. It is found that Arg-17 is likely to be of importance in order to understand the way BPTI binds on trypsin.     

Spectral and statistical analysis of fluctuations in the SOL and diverted plasmas of the Uragan-3M torsatron

In the l = 3/m = 9 Uragan-3M (U3-M) torsatron (R ₀ = 1 m, ā ≈ 0.12 m, ι(ā)/2π ～ 0.3) with an open helical divertor and a plasma produced and heated by RF fields (ω ? ω_ci), studies of low frequency (5–100 kHz) density and potential fluctuations in the SOL plasma and in the diverted plasma flows (DPFs), have been carried out. It is shown, that in the SOL to more (less) distantly located points relative to the last closed magnetic surface, higher (lower) frequency fluctuations are inherent. Such a spectral splitting in two sub-ranges occurs in the DPFs too. A peculiarity of the spatial distribution of DPF fluctuation spectra is that lower frequency fluctuations dominate on the ion toroidal B × ?B drift side. During L-H-like transition in U-3M simultaneously with strong E _r shear formation, a suppression of lower frequency fluctuations and a decrease of local radial turbulent particle flux take place. Results are presented of investigation of plasma density fluctuations in SOL with the use of probability distribution function (PDF) analysis. Evaluations of skewness and kurtosis of fluctuations have been made. The analysis of I _s fluctuations in DPF have been carried out in a similar way.     

Influence of macroinvertebrates and macrophytes on waterfowl utilization of wetlands in the Prairie Pothole Region of northwestern Wisconsin

Waterfowl and limnological data were monitored on Waterfowl Production Area (WPA) wetlands in northwestern Wisconsin over a 6-yr period (1983–88) to determine the impact of macroinvertebrates and macrophytes on waterfowl utilization. Interrelationships between limnological conditions and Waterfowl Breeding Pair Densities (BPDs reported as pairs/ha water surface) were analyzed using correlation and general linear model analysis techniques.Annual changes in waterfowl BPDs differed between wetlands according to differences in the structure of macrophyte communities and basin morphometry. The strength of associations differed between the two dominant waterfowl species. In a wetland dominated by dense stands of submersed vegetation, annual fluctuations in blue-winged teal (Anas discors) BPDs corresponded directly with changes in macrophyte biomass, but not with changes in macroinvertebrate density. In a nearby less densely vegetated wetland of similar water chemistry and trophic status, fluctuations in teal BPDs corresponded directly with changes in macroinvertebrate density, but not with changes in macrophyte biomass. These associations occurred despite a significant positive correlation between macroinvertebrates and macrophyte biomass in the latter habitat. Annual fluctuations in mallard (Anas platyrhynchos) BPDs were not correlated significantly with either macrophyte biomass or macroinvertebrate density in either wetland.     

Anomalous permeability and stability characteristics of erythrocytes in non-electrolyte media

The permeability characteristics of the erythrocyte membrane were critically evaluated in electrolyte and non-electrolyte (sucrose) media by ion-selective electrodes and radioactive polyol fluxes as well as by the novel technique of osmometry. K+ efflux demonstrated a linear osmotic susceptibility distinct from Na+ influx upon incubation in NaCl media of various tonicities. In non-electrolyte media, acidification of the medium, large fluxes of K+, sucrose and even haemoglobin (as manifest by hypertonic disruption) were consistent with enhanced porosity of the bilayer due to the field created by surface charge density leading to density fluctuations in the bilayer.     

Endogenous oscillator controls surface density of mating receptors in Chlamydomonas eugametos

Summary We describe a circadian rhythm in the surface density of receptors that play a dominant role in the mating process of the unicellular green alga Chlamydomonas eugametos.These receptors — called agglutinins — are large glycoproteins extrinsically bound to the membrane of gamete flagella. We found circadian fluctuations in their density. Since inhibition of protein synthesis affected the agglutinin density without a lag period at any time,we conclude that the density was dependent on de novo synthesis and that the fluctuations in density are caused by circadian oscillations in the rate of agglutinin synthesis. This phenomenon evidently underlies the pronounced endogenous rhythm in mating competence that we described previously (Demets et al. 1987). Finally, we speculate on the nature of the time keeping mechanism that is generating these rhythmic events.     

Large-scale quantitative analysis of sources of variation in the actin polymerization-based movement of Listeria monocytogenes

During the actin polymerization-based movement of Listeria monocytogenes, individual bacteria are rapidly propelled through the host cell cytoplasm by the growth of a filamentous actin tail. The rate of propulsion varies significantly among individuals and over time. To study this variation, we used a high-throughput tracking technique to record the movement of a large number (approximately 7900) of bacteria in Xenopus frog egg extract. Most bacteria (70%) appeared to maintain an individual characteristic speed over several minutes, suggesting that the major source of variation in average speed is intrinsic to the bacterium. Thirty percent of bacteria had significant changes in speed over time spans of a few minutes, including 17% that appeared to collide with obstacles and 13% that moved with a significant periodic component. For the latter, the peak frequency was proportional to speed, suggesting a mechanism with a fixed spatial scale of approximately 0.6 bacterial length. Near the rear of the bacterium, temporal fluctuations in actin density were positively correlated with fluctuations in speed, whereas near the front the correlation was negative. A comparison of the performance of linear models that predict motion given actin density suggests that the mechanism has a history of 5-10 s, and that fluctuations in actin density near the front of the bacteria contain more predictive information than the rear. Our results are consistent with physical models where bacterial speed is governed by the rate of dissociation of bonds between the bacterial surface and the actin tail, and individual variation is determined by long-lived intrinsic variability in bacterial surface properties.     

Structure of nuclear pre-mRNA. IX. Hybridization of double-stranded portions of pre-mRNA with excess mRNA

The hybridization of double-stranded regions of pre-mRNA from mouse Ehrlich ascites carcinoma cells, rabbit bone marrow cells and primary culture of rabbit kidney cells with an excess of total poly(A)+-mRNA of mouse or rabbit globin mRNA respectively was studied. The hybrids were detected as RNAase-stable acid precipitable material or by adsorbtion of the hybrid complexes of poly(U)-sepharose. The sizes of the hybrid complementary sequences and their thermal stability were estimated.     

Correlated conformational fluctuations during enzymatic catalysis: Implications for catalytic rate enhancement

Correlated enzymatic conformational fluctuations are shown to contribute to the rate of enhancement achieved during catalysis. Cytidine deaminase serves as a model system. Crystallographic temperature factor data for this enzyme complexed with substrate analog, transition-state analog, and product are available, thereby establishing a measure of atomic scale conformational fluctuations along the (approximate) reaction coordinate. First, a neural network-based algorithm is used to visualize the decreased conformational fluctuations at the transition state. Second, a dynamic diffusion equation along the reaction coordinate is solved and shows that the flux velocity through the associated enzymatic conformation space is greatest at the transition state. These results suggest (1) that there are both dynamic and energetic restrictions to conformational fluctuations at the transition state, (2) that enzymatic catalysis occurs on a fluctuating potential energy surface, and (3) a form for the potential energy. The Michaelis-Menten equations are modified to describe catalysis on this fluctuating potential energy profile, leading to enhanced catalytic rates when fluctuations along the reaction coordinate are appropriately correlated. This represents a dynamic tuning of the enzyme for maximally effective transformation of the ES complex into EP.     

Structure and sorption properties of regenerated hemosorbent SCN

Stepwise desorption of protein components from the hemosorbent SCN after hemoperfusion was studied, and their molecular weights were determined. It was found that hard alkaline regeneration and oxidation with nitric acid do not affect the hemosorbent structure, composition and potentiometric characteristics. The regenerated hemosorbent can be successfully used for adsorbtion of extraneous low- and medium-molecular weight components to clarify microbiological liquids.     

How Nature Modulates Inherent Fluctuations for Biological Self-Organization – The Case of Membrane Fusion

Biological systems in nano-scale, due to the weak electrostatic interactions and structural connectivity therein, are flexible so that they undergo conformational transition subject to thermal fluctuations and external noises. In the presence of barriers, nature utilizes the fluctuations to give rise to self-organization, typically accompanied by conformational transitions. In two opposing membranes with like-charges, the cooperative coupling between the undulation and charge fluctuations give rise to a dynamic instability to spontaneous growth of the in-phase membrane undulation, and thus a great reduction of the energy barrier to fusion. The multivalent counter-ions, the Ca²⁺ for example, enhance the necessary charge density fluctuation leading to surface charge inversion and overcondensation.     

田鼠种群波动的原因和时间

本文总结了橙腹田鼠（Microtus ochrogaster）和草原田鼠（M．permsylvanicus）25年的种群统计学研究结果和结论。探讨了田鼠种群波动周期性、诱发种群波动以及导致波动期间峰值变异的因素。并对种群存活值和繁殖活动的作用进行了分析和评价。根据两种田鼠种群波动周期性、波动峰值出现的时间以及特定年份峰值的高度等特征,证明两物种波动均具有不稳定性。两种田鼠存活值的变化是由特定年份是否发生波动以及波动峰值出现的时间决定。增加初始阶段的种群密度及时间长度是造成两种动物种群波动峰值不同的主要原因。橙腹田鼠种群停止增长的原因是存活值降低,而草原田鼠则是繁殖活动减少。据推测,与种群波动初始密度相关的种群死亡率的差异是由捕食者的净效应（Net effect）决定的,调控两种群密度的因素均为非密度的其它生态学因子。由于特定年份田鼠种群捕食压力的不确定性,导致了橙腹田鼠和草原田鼠种群波动的不稳定性。     

Protein dynamics in a family of laboratory evolved thermophilic enzymes

Molecular dynamics simulations were employed to study how protein solution structure and dynamics are affected by adaptation to high temperature. Simulations were carried out on a para-nitrobenzyl esterase (484 residues) and two thermostable variants that were generated by laboratory evolution. Although these variants display much higher melting temperatures than wild-type (up to 18 degrees C higher) they are both >97% identical in sequence to the wild-type. In simulations at 300 K the thermostable variants remain closer to their crystal structures than wild-type. However, they also display increased fluctuations about their time-averaged structures. Additionally, both variants show a small but significant increase in radius of gyration relative to wild-type. The vibrational density of states was calculated for each of the esterases. While the density of states profiles are similar overall, both thermostable mutants show increased populations of the very lowest frequency modes (<10 cm(-1)), with the more stable mutant showing the larger increase. This indicates that the thermally stable variants experience increased concerted motions relative to wild-type. Taken together, these data suggest that adaptation for high temperature stability has resulted in a restriction of large deviations from the native state and a corresponding increase in smaller scale fluctuations about the native state. These fluctuations contribute to entropy and hence to the stability of the native state. The largest changes in localized dynamics occur in surface loops, while other regions, particularly the active site residues, remain essentially unchanged. Several mutations, most notably L313F and H322Y in variant 8G8, are in the region showing the largest increase in fluctuations, suggesting that these mutations confer more flexibility to the loops. As a validation of our simulations, the fluctuations of Trp102 were examined in detail, and compared with Trp102 phosphorescence lifetimes that were previously measured. Consistent with expectations from the theory of phosphorescence, an inverse correlation between out-of-plane fluctuations on the picosecond time scale and phosphorescence lifetime was observed.