Shapes of curves of pH-dependence of reactions

A simple case is considered in which the rate of a two-step reaction depends on pH because the intermediate formed in the first step has to gain (or lose) a proton before it can react in the second step, and in which the rate-determining step therefore changes with pH. The curves of reaction rate against pH are shown to be symmetrical, and the sharpest peak possible has a width at half its height of 1.53pH units, i.e. of 2log(3+2 radical2). Any particular curve for this situation proves to be identical with a curve that could be generated for the pH-dependence of a single-step reaction in which the rate is proportional to the concentration of a particular ionic form of a reactant. Curves for the latter situation, however, can have forms impossible for the former case in which the rate-determining step changes, but only if the protonations that activate and deactivate the reactant are co-operative. The peak can then become even sharper, and its width at half its height can fall to 1.14pH units, i.e. to 2log(2+ radical3).     

An analysis of non-bonded energy of proteins

Non-bonded energy of 16 proteins was calculated using the atomic co-ordinates obtained by X-ray crystallography. The curve of total energy against the number of atoms in proteins is approximately linear with a slight concaved shape. According to a linear equation to fit the curve, the extrapolated length of a polypeptide chain of a globular shape is expected to be 18 residues, which corresponds conceivably to an approximate size of nucleus for a folding of the polypeptide chain. Contributions from short-range and medium-range energies are always much greater than those from long-range energy for all the proteins and there seems to exist a change of each contribution in a range from 1200 to 1700 atoms. The energies with a lag less than four residues are a major part of the total energy and the contribution of energy from main-chain atoms is considerably higher than that from side-chain atoms. Side-chain atoms of a residue have a tendency to interact more strongly with main-chain atoms of N-terminal, than with those of C-terminal side of the residue, indicating asymmetry of the interaction in a protein. Amino acid residues in proteins may be divided into three groups by the order of strength of average energy. The first group exhibiting strong interaction consists mainly of hydrophobic amino acids and the third group consists of hydrophilic ones corresponding to the location in a protein molecule. Cys, val, leu and met are important for medium-range and long-range energies; gly and ala for medium-range energy; ile, trp, phe, tyr and arg for long-range energy. One simple application of the average energy of amino acid residues is illustrated to estimate local energy of a segment of nine residues given by a protein sequence. There is a good correlation between the curve computed by the average energy and the experimental curve for myoglobin.     

Findings of the importance of dietetic fiber in the regulation of cholesterolemia

We studied the variations of some lipid indexes in cholesterolemia, lipidemia and triglycerides in relation to a diet rich in bulk. Seven adult male subjects in good health were tested for cholesterol, lipid and triglycerides before and after a two week period of a dialy diet of 40 g of whole wheat bran containing about 10g of fiber. At the some time the lipid, cholesterol and triglycerides limits were measured in 3 groups of rabbits. The first groups was subjected to a normal diet, the second group was subjected to a diet of bleached flour and vitamins, and the third group had a diet similar to the second group but with the addition of 40 g whole wheat bran. The results of our investigation demonstrated a statistically significant decrease of cholesterol lipid and triglycerides levels the special diets at the some time we did not see in increase in the third group of rabbit. Our study seems to indicate that dietetic fibers exercise a vital role in regulating the concentration of plasma lipids even if the subject continues to consume a diet rich in fats. From this we see to importance of the dietetic fibers as a protective factor in the prevention of ateriosclerotic.     

Isolation of domains of the plasma membrane of hepatocytes

Several recent studies have demonstrated the ability of techniques based on immunoadsorption to selectively isolate specialized subregions of membranes, termed domains, which are derived from a larger more complex parent membrane like the plasma membrane. The immunoadsorbent is directed against a specific antigen that resides exclusively or predominantly in the membrane domain to be isolated. Thus, a monospecific antibody to the domain-specific antigen is required. In the present study we developed a method employing a modified immunoblotting strategy which could utilize polyspecific antibodies to isolate membrane vesicles derived from a specific membrane domain of the hepatocyte plasma membrane. We also used specific cell surface labeling of the hepatocyte plasma membrane by lactoperoxidase-catalyzed iodination at 4 degrees C and preparation of different sized vesicles by sonication to facilitate isolation of the specific domain. For this study, polyspecific antisera were raised in goats against a membrane fraction, denoted N2u, which is enriched in bile canalicular proteins. This antiserum recognizes, among other antigens, a 110,000 Mr polypeptide previously shown to be localized in the bile canaliculus (J. Cook et al. (1983) J. Cell. Biol. 97, 1823-1833). A monospecific antiserum was raised in rabbits against the rat hepatocyte asialoglycoprotein receptor, a sinusoidal domain-specific set of glycoproteins whose major form has a Mr of 43,000. These antisera were each coupled indirectly to different pieces of nitrocellulose by the immunoblotting protocol and were used to isolate membrane vesicles from a crude extract of liver plasma membrane prepared by sonication. The ratio of iodinated asialoglycoprotein receptor to the 110,000 Mr polypeptide in vesicles isolated by the affinity nitrocellulose immunoadsorbent method indicate a 10- to 15-fold enrichment of sinusoidal-derived vesicles relative to bile canalicular-derived membrane vesicles. These results show that the affinity nitrocellulose immunoadsorbent method can be used to isolate domain-specific vesicles. Further, the affinity immunoadsorbent method described here for the isolation of domains of the plasma membrane is an integrative one allowing isolation of vesicles present in relatively small concentration in crude cell extracts and it requires minimal ultracentrifugation time.     

Stages of the cerebral mechanisms of deceptive responses

Cerebral mechanisms of perceiving and telling lies were studied by recording event-related potentials (ERPs) both after an actual deceptive response and during the time interval when the subject decided to tell a lie. Ten healthy volunteers participated in the study. The test consisted of their playing a game against a computer. The subjects could choose between deceptive and truthful answers so as to win the game. The subjects gave a deceptive answer intentionally, the structure of the test ensuring equal numbers of deceptive and truthful answers. The relaxation times in the cases of truthful and deceptive answers did not differ significantly from each other. The comparison of ERPs accompanying deceptive and truthful answers showed the existence of a negativity with a latent period of 90 ms in the regions of the right frontal, central, and right parietal derivations. This negativity indicated that the brain reacted to a deceptive answer even if this a priori “erroneous” act ensured reaching the goal and, in this sense, was subjectively relevant. In terms of the cerebral error detector mechanism, this phenomenon may be regarded as a special case of a general response of the brain to giving an incorrect (deceptive) answer, rather than a response to a lie per se. The interval of time when, presumably, the decision on a deceptive answer was being made was found to contain the late positive component P540, which is most likely to be involved in the preparation of the deceptive answer and the intention to tell a lie.     

Thermostability of Proteases and Lipases from a Number of Species of Psychrotrophic Bacteria of Dairy Origin

Proteases and lipases from a number of different species of psychrotrophic bacteria isolated from dairy products are resistant to heat treatments of 77°C for a holding time of 17 s and 140°C for a holding time of 5s. A further treatment combining the 77°C for 17 s heating process with a heat treatment involving a temperature of 55°C for 1 h was also examined. The proteases and, to a lesser extent, the lipases were also resistant to the treatments in combination. The resistance of proteases and lipases to inactivation by a treatment of 55°C for 1 h varied between and within species. Therefore, this treatment may not have a widespread practical application.     

Possible mechanisms of implementation of toxic potential of lipopolysaccharides of pathogenic bacteria

The significance of variability of biological properties of lipopolysaccharides (LPS) is discussed in the paper within the pathogenesis of infectious process. On the basis of an analysis of published data and of results of independent research of two microorganisms (Yersinia pestis and Francisella tularensis) a conclusion is made on that a biologically inert LPS form (with a weak cytokine-inducing ability, apirogenicity and non-toxicity etc.) is typical of highly pathogenic bacteria. It is suggested that the above phenomenon is biologically expedient. Presumably, the inert LPS transforms to the active form inside a sensitive host and, according to an infection stage, each of them being functionally significant. It is the inert status of LPS that enables the pathogens, at the initial stages, to surmount freely the humoral and cell barriers of host. As the infection progressively aggravates and the proliferation of bacteria modifies itself due to LPS micro- and macroorganisms, its chemical structure and biopolymer conformation change. Both modification mechanisms enhance the LPS toxic potential. In case of a sensitive host, such variations transform the biologically inert LPS into a toxically active form with its function of endotoxin being realized. There is no LPS modification in a host insensitive to such infection, which entails either recovery or prolonged persistence of the pathogen inside the microorganism.     

Application of the function of the number of states of dynamic system to investigation of electroencephalographic reaction to photostimulation

A new way of EEG analysis is developed, which makes it possible to reveal a reaction of the brain cortex to external stimulation at a low response level. The method is based on the creation of an image of the EEG curve in the discrete phase space and comparison of a current condition with the conditions from a certain basic set. The strength of reaction to external stimulation is defined by the incidence of conditions novel with respect to the basic ones.     

Tracer measurements of non-equilibrium volumes of distribution

According to conventional theory the product of the transport flowrate and the mean transit time of a tracer through a system yields the equilibrium volume of distribution for the tracer, regardless of tracer kinetics or space geometry. Experimental results do not support this notion. The influence of measurement time on the volume measured with a bolus technique is addressed using systems theory to analyze a tissue-impedance form of the Sangren-Sheppard model. Assymptotic solutions show that the volume estimates are governed by a time constant, tau, related to diffusion in the tissue, to tissue capacity, and to wall permeability, and by a dimensionless ratio, f, describing a relation of tau to vascular transport time. A third parameter, g, describing the relative contributions to overall resistance to diffusion of effective permeability and of limited diffusivity in the tissue, is shown to be of less importance. The derived tau is similar to but not equivalent to the often cited "characteristic time". The "equilibrium" volume of distribution is defined as that which would be measured if equilibrium were allowed to establish. The "non-equilibrium" volume of distribution is defined as that which would be measured given finite times and is shown to approach the "equilibrium" volume as such times increase. Tracer equilibration is not required to accurately measure the "equilibrium" volume. When there is no flow limitation (f much less than 1) a measurement time of tau (plus vascular transit time) would yield a "non-equilibrium" volume only 33% of the "equilibrium" volume; a time of 2 tau would yield 55%; a time of 10 tau would yield effectively the total equilibrium volume. Finite diffusivity in tissue and permeability restrictions can have significant effects on the proportion of the volume measured.     

The Effect of Conflicting Pressures on the Evolution of Division of Labor

Within nature, many groups exhibit division of labor. Individuals in these groups are under seemingly antagonistic pressures to perform the task most directly beneficial to themselves and to potentially perform a less desirable task to ensure the success of the group. Performing experiments to study how these pressures interact in an evolutionary context is challenging with organic systems because of long generation times and difficulties related to group propagation and fine-grained control of within-group and between-group pressures. Here, we use groups of digital organisms (i.e., self-replicating computer programs) to explore how populations respond to antagonistic multilevel selection pressures. Specifically, we impose a within-group pressure to perform a highly-rewarded role and a between-group pressure to perform a diverse suite of roles. Thus, individuals specializing on highly-rewarded roles will have a within-group advantage, but groups of such specialists have a between-group disadvantage. We find that digital groups could evolve to be either single-lineage or multi-lineage, depending on experimental parameters. These group compositions are reminiscent of different kinds of major evolutionary transitions that occur within nature, where either relatives divide labor (fraternal transitions) or multiple different organisms coordinate activities to form a higher-level individual (egalitarian transitions). Regardless of group composition, organisms embraced phenotypic plasticity as a means for genetically similar individuals to perform different roles. Additionally, in multi-lineage groups, organisms from lineages performing highly-rewarded roles also employed reproductive restraint to ensure successful coexistence with organisms from other lineages.     

Ultrastructure of bovine blastocysts following cryopreservation: effect of method of blastocyst production

The objective of this study was to describe the ultrastructure of blastocysts derived by in vivo and in vitro methods and to investigate how the morphology is affected by exposure to cryoprotectant (10% glycerol) or cryopreservation by conventional slow freezing. In vivo derived blastocysts were characterized by a narrow perivitelline space (PvS), a continuous cover of numerous stacked microvilli (MV) on the plasma membrane, a well-defined system of cell-to-cell coupling and a large population of round or elongated mitochondria with numerous transverse cristae. Exposure of these blastocysts to cryoprotectant was manifested by shrinkage of the blastocysts and swelling of the mitochondria. Cryopreservation resulted in further shrinkage, damage to the MV, and accumulation of cellular debris. In comparison, the in vitro matured (IVM)/in vitro fertilized (IVF) in vivo cultured blastocysts displayed a wider PvS; they appeared to possess less MV and all blastocysts displayed some cellular debris in their PvS. There was also a decrease in the number of junctional contacts between the trophoblastic cells. The reaction of these blastocysts to exposure to cryoprotectant was similar to that of the in vivo derived blastocysts. However, they appeared to be more susceptible to cryopreservation. The totally in vitro produced (IVP) blastocysts displayed a wider PvS, no stacking of the MV, increased numbers of lipid droplets and a further reduction in the junctional contacts between trophoblastic cells. The IVP blastocysts sustained breakage of the zona pellucida on exposure to cryoprotectant and were extremely sensitive to cryopreservation, losing all cell structure and organization. The findings of the present study indicate that in vivo derived blastocysts possess certain structural characteristics that confer a greater tolerance on them to exposure to cryoprotectant and cryopreservation.     

Mechanism of allosteric effects of ATP on the kinetics of P-type ATPases

The roles of allosteric effects of ATP and protein oligomerisation in the mechanisms of P-type ATPases belong to the most controversial and least well understood topics in the field. Recent crystal structural and kinetic data, however, now allow certain hypotheses to be definitely excluded and consistent hypotheses to be developed. The aim of this review is to critically discuss recent results and, in the light of them, to present a set of conclusions which could form the basis of future research. The major conclusions are: (1) at saturating ATP concentrations P-type ATPases function as monomeric enzymes, (2) the catalytic units of P-type ATPases only possess a single ATP binding site, (3) at non-saturating ATP concentrations P-type ATPases exist as diprotomeric (or higher oligomeric) complexes, (4) protein–protein interactions within a diprotomeric complex enhances the enzymes’ ATP binding affinity, (5) ATP binding to both protomers within a diprotomeric complex causes it to dissociate into two separate monomers. The physiological role of protein–protein interactions within a diprotomer may be to enhance ATP binding affinity so as to scavenge ATP and maximize the ion pumping rate under hypoxic or anoxic conditions. For the first time a structural basis for the well-known ATP allosteric acceleration of the E2 → E1 transition is presented. This is considered to be due to a minimization of steric hindrance between neighbouring protomers because of the ability of ATP to induce a compact conformation of the enzymes’ cytoplasmic domains.     

北半球500hPa高度场遥相关指数对我国草地螟成虫物候期与幼虫发生面积的影响

北半球500hPa高度场的遥相关指数(遥相关指数)是一个综合了气温,降水以及气压等众多气象因素的气候变化指数,很多生物学现象都与其有关;为了探究遥相关指数是否对我国的草地螟的发生危害有影响,作者分析了我国草地螟Loxostege sticticalisL.成虫物候期及幼虫发生面积的变化趋势及两者与遥相关指数的相关关系,结果表明:我国草地螟成虫始见期年际间有显著提前的趋势,平均每年提前0.40d,与1—5月极地-欧亚型遥相关指数均值(POL1-5)相关性最大;终见期有显著延后的趋势,平均每年延后1.31d,与6—10月东太平洋型遥相关指数均值(EP-NP6-10)相关性最大;全国草地螟幼虫发生面积年际间有显著增长的趋势,平均每年增长57.07khm2,与1—10月东大西洋型遥相关指数均值(EA1-10)相关性最大。     

Statistical fluctuations in processes of gene expression regulation: consideration of the problem from point of view of statistical mechanics

The regulation of gene expression is a basic problem of biology. In some cases, the gene activity is regulated by specific binding of regulatory proteins to DNA. In terms of statistical mechanics, this binding is described as the process of adsorption of ligands on the one-dimensional lattice and has a probability nature. As a random physical process, the adsorption of regulatory proteins on DNA introduces a noise to the regulation of gene activity. We derived equations, which make it possible to estimate this noise in the case of the binding of the lac repressor to the operator and showed that these estimates correspond to experimental data. Many ligands are able to bind nonspecifically to DNA. Nonspecific binding is characterized by a lesser equilibrium constant but a greater number of binding sites on the DNA, as compared with specific binding. Relations are presented, which enable one to estimate the probability of the binding of a ligand on a specific site and on nonspecific sites on DNA. The competition between specific and nonspecific binding of regulatory proteins plays a great role in the regulation of gene activity. Similar to the one-dimensional "lattice gas" of particles, ligands adsorbed on DNA produce "one-dimensional" pressure on proteins located at the termini of free regions of DNA. This pressure, an analog of osmotic pressure, may be of importance in processes leading to changes in chromatin structure and activation of gene expression.     

Certification of Probability of Sterilization of Liquid by Filtration

Four types of hydrosol filters, two reusable (diatomaceous cylinder and fritted-glass funnel) and two disposable (asbestos pad and membrane filter) were challenged with a heavy bacterial suspension to assess their ability to produce sterile filtrates. Two of the four diatomaceous earth filters, the four fritted-glass funnels, and all of the asbestos pads tested generally gave sterile filtrates. However, only one type of filter, one of the membranes in its manufacturer's own holder, consistently gave sterile filtrates. The two other types of membranes usually gave sterile filtrates if tested in one manufacturer's holder, but all types invariably gave contaminated filtrates when tested in another manufacturer's holder. Contaminated filtrates were generally attributed to a poor reusable filter or to a faulty holder used with a disposable filter. If a high degree of certainty is required for sterile heat-labile filtrate, it is suggested that the liquid be passed through two or more filters in a previously tested and proven system.     

Analysis of Protein Structural Motifs in Terms of Sets of Codes Representing Local Structures

An amino acid sequence pattern conserved among a family of proteins is called motif. It is usually related to the specific function of the family. On the other hand, functions of proteins are realized through their 3D structures. Specific local structures, called structural motifs, are considered as related to their functions. However, searching for common structural motifs in different proteins is much more difficult than for common sequence motifs. We are attempting in this study to convert the information about the structural motifs into a set of one-dimensional digital strings, i.e., a set of codes, to compare them more easily by computer and to investigate their relationship to functions more quantitatively. By applying the Delaunay tessellation to a 3D structure of a protein, we can assign each local structure to a unique code that is defined so as to reflect its structural feature. Since a structural motif is defined as a set of the local structures in this paper, the structural motif is represented by a set of the codes. In order to examine the ability of the set of the codes to distinguish differences among the sets of local structures with a given PROSITE pattern that contain both true and false positives, we clustered them by introducing a similarity measure among the set of the codes. The obtained clustering shows a good agreement with other results by direct structural comparison methods such as a superposition method. The structural motifs in homologous proteins are also properly clustered according to their sources. These results suggest that the structural motifs can be well characterized by these sets of the codes, and that the method can be utilized in comparing structural motifs and relating them with function.     

Modeling of time-dependent force response of fingertip to dynamic loading

An extended exposure to repeated loading on fingertip has been associated to many vascular, sensorineural, and musculoskeletal disorders in the fingers, such as carpal tunnel syndrome, hand-arm vibration syndrome, and flexor tenosynovitis. A better understanding of the pathomechanics of these sensorineural and vascular diseases in fingers requires a formulation of a biomechanical model of the fingertips and analyses to predict the mechanical responses of the soft tissues to dynamic loading. In the present study, a model based on finite element techniques has been developed to simulate the mechanical responses of the fingertips to dynamic loading. The proposed model is two-dimensional and incorporates the essential anatomical structures of a finger: skin, subcutaneous tissue, bone, and nail. The skin tissue is assumed to be hyperelastic and viscoelastic. The subcutaneous tissue was considered to be a nonlinear, biphasic material composed of a hyperelastic solid and an invicid fluid, while its hydraulic permeability was considered to be deformation dependent. Two series of numerical tests were performed using the proposed finger tip model to: (a) simulate the responses of the fingertip to repeated loading, where the contact plate was assumed to be fixed, and the bone within the fingertip was subjected to a prescribed sinusoidal displacement in vertical direction; (b) simulate the force response of the fingertip in a single keystroke, where the keyboard was composed of a hard plastic keycap, a rigid support block, and a nonlinear spring. The time-dependent behavior of the fingertip under dynamic loading was derived. The model predictions of the time-histories of force response of the fingertip and the phenomenon of fingertip separation from the contacting plate during cyclic loading agree well with the reported experimental observations.     

Comparison of models of insulin release

A model for insulin secretion with a storage and a labile compartment, as well as a provisionary factor, is combined with a signal model in which the signal can be the difference between an excitation and an inhibition, or the difference in concentrations inside and outside some cell components. The model, using a single set of values for the parameters, accounts in a semiquantitative manner for all of the regularly appearing features of the insulin secretion from thein vitro perfused pancreas to a wide range of patterns of glucose and tolbutamide stimulation. Among the features which can be accounted for are: early and late secretion of insulin as a function of glucose in terms of a single parameter; the apparent depletion and recovery during a pulsed pattern of stimulation by tolbutamide; the hypersecretion following a short period of rest during a prolonged stimulation by glucose; the negative spike which occurs when the concentration of glucose, which has been maintained for a period of time, is suddenly reduced to a lower level; and the appropriate responses to slow and fast ramp functions of glucose concentration.     

Discounting of sequences of delayed rewards of different amounts

The main purpose of this study was to determine whether the magnitude effect is present in cases where delayed sequences of rewards are discounted. The magnitude effect refers to the inverse relationship between the amount of a reward and the steepness of temporal discounting. This study was conducted with a computer program to estimate the indifference points, which served as indicators of the present subjective value of delayed sequences of small and large rewards. In the indifference point the subjective value of a single, immediate reward was equal to the subjective value of the delayed sequence (or to the value of a single delayed reward). As a control condition, we added an experimental task involving choices between single immediate and single delayed rewards. The experiment showed that the sequences of large rewards are discounted less steeply than are the sequences of small rewards. This finding suggests that the magnitude effect is present within the delayed sequences of rewards. In addition, when outcomes are relatively large, the results suggest that a single reward is discounted less steeply than the sequence of a total nominal value equal to this single reward. However, for relatively small rewards, the difference is not statistically significant. The less steep discounting of sequences of large rewards may explain the reward-bundling effect, which refers to less steep discounting of longer sequences than of shorter ones: longer sequences usually have greater overall nominal value. The present study was conducted on hypothetical rewards, and the results should be validated using real rewards.