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).     

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.     

Simulations of action of DNA topoisomerases to investigate boundaries and shapes of spaces of knots

The configuration space available to randomly cyclized polymers is divided into subspaces accessible to individual knot types. A phantom chain utilized in numerical simulations of polymers can explore all subspaces, whereas a real closed chain forming a figure-of-eight knot, for example, is confined to a subspace corresponding to this knot type only. One can conceptually compare the assembly of configuration spaces of various knot types to a complex foam where individual cells delimit the configuration space available to a given knot type. Neighboring cells in the foam harbor knots that can be converted into each other by just one intersegmental passage. Such a segment-segment passage occurring at the level of knotted configurations corresponds to a passage through the interface between neighboring cells in the foamy knot space. Using a DNA topoisomerase-inspired simulation approach we characterize here the effective interface area between neighboring knot spaces as well as the surface-to-volume ratio of individual knot spaces. These results provide a reference system required for better understanding mechanisms of action of various DNA topoisomerases.     

Importance of preservation of biophysical organization of isolated mitochondria for revealing physiological regulation of their functions

A method has been elaborated that preserves the mitochondrial-reticular network in lymphocytes in composition to the physiological one. Physiologicalby the immobilization of a blood smear on glass and its subsequent incubation in a medium closeresponses of respiration to excitation in the ition of early responses of ions. The recogn organism are well pronounced on these preparat mitochondria to pathogenic agents in the organism is a timely problem of basic and medicinal e- investigations since they play a leading role in the development of pathological states.     

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.     

A neurobiophysical interpretation of certain aspects of the problem of risks

A situation is considered in which an individual is given an opportunity to risk a certain amount of money or goods in order to gain a larger amount, providing the result of some uncertain event proves favorable. A neural mechanism is introduced in which the probability of success can be ordered according to its value. The response to a situation thus becomes dependent upon the probability. An expression is then derived for the amount that would be risked in terms of the probability and of the amount that would be gained in the event of a successful outcome. Similar expressions are obtained for the case of insurance against loss. Results of questionnaires indicate that individuals can be classified according to their pattern of behavior in the situation considered. The various types can be most easily recognized when a plot is made of the relative amount risked against the probability of success. In a general way, these types can be understood in terms of the equations derived.     

Consequences of postural changes and removal of vestibular inputs on the movement of air in and out of the lungs of conscious felines.

A variety of experimental approaches in human subjects and animal models established that the vestibular system contributes to regulation of respiration. In cats, the surgical elimination of labyrinthine signals produced changes in the spontaneous activity and posturally related responses of a number of respiratory muscles. However, these effects were complex and sometimes varied between muscle compartments, such that the physiological role of vestibulo-respiratory responses is unclear. The present study determined the functional significance of vestibulo-respiratory influences by examining the consequences of a bilateral labyrinthectomy on breathing rate and the pressure, volume, and flow rate of air exchanged during inspiration and expiration as body orientation with respect to gravity was altered. Data were collected from conscious adult cats acclimated to breathing through a facemask connected to a pneuomotach during 60 degrees head-up pitch and ear-down roll body rotations. Removal of vestibular inputs resulted in a 15% reduction in breathing rate, a 13% decrease in minute ventilation, a 16% decrease in maximal inspiratory airflow rate, and a 14% decrease in the maximal expiratory airflow rate measured when the animals were in the prone position. However, the lesions did not appreciably affect phasic changes in airflow parameters related to alterations in posture. These results suggest that the role of the vestibular system in the control of breathing is to modify baseline respiratory parameters in proportion to the general intensity of ongoing movements, and not to rapidly alter ventilation in accordance with body position.     

Features of the replicon of plasmid pAM10.6 of Pseudomonas fluorescens

We describe features of the basic replicon of the 10.6-kb medium-copy-number plasmid pAM10.6. pAM10.6 was able to replicate in various Pseudomonas strains but was maintained in Escherichia coli only after the p15A origin of replication was inserted. Deletion analysis suggests that the pAM10.6 origin of replication is located in a 0.5-kb region that includes inverted and direct repeats upstream of the repA gene. RepA (204 aa) has a clear homology to plasmid replication proteins of some other gram-negative bacteria. The pas (plasmid addiction system) (genes encoded in the region of 480-bp) stabilizes plasmid maintenance in P. putida cells under nonselective conditions for at least 200 generations. A 3.75-kb PstI fragment of pAM10.6 joined to a Km(r) gene was shown to be a minimal plasmid unit maintained in P. putida as a monomer. Further deletions of this 3.75-kb fragment caused a drive to form stable head-to-tail dimeric plasmids in P. putida.     

Effect of the Osmotic Pressure of the Growth Medium on fabB Mutants of Escherichia coli

Temperature-sensitive, unsaturated fatty acid (fabB) auxotrophs of Escherichia coli can grow at the restrictive temperature in the absence of unsaturated fatty acid in a medium with a high osmotic pressure. If a mutant culture was starved for unsaturated fatty acids and harvested just before the lysis started, the fatty acid composition of the cells was the same as that of cells grown until late log phase in a high-osmotic medium. Evidence is presented that the in vivo unsaturated fatty acid biosynthesis is significantly increased in a high osmotic medium. The increase is probably due to a partial activation of the temperature-sensitive fabB product. Besides the stimulation of the temperature-sensitive fabB product, a minimal osmotic pressure of the medium appeared to be necessary to allow growth of cells containing lipids with a changed fatty acid composition. fabA mutants are unable to grow in a high-osmotic medium in the absence of unsaturated fatty acids. No increase in the in vivo unsaturated fatty acid biosynthesis could be detected in the temperature-sensitive fabA mutants.     

The isolation of a dimer of gene 8 protein of bacteriophage fd

Gene 8 protein was isolated in a highly aggregated form in aqueous solution from purified fd bacteriophage. Gene 8 protein was dissociated to a stable dimeric form in the presence of 10 mm-sodium deoxycholate. The dimer obtained in the presence of the detergent was stable to further dissociation by sodium dodecyl sulfate, and was estimated to have a 51% helical content on the basis of circular dichroism data. When the deoxycholate was removed, the dimers were observed by electron microscopy to self-associate to complex string-like structures. The dimer obtained is thought to be a result of one of the interactions between adjacent monomers in the model proposed by Marvin & Wachtel (1975).     

Effect of conditioning polarization of soma of giant neurons of mollusks on the mechanism of action-potential generation

To ascertain the properties of an excitable membrane of the soma of giant neurons of mollusks, experiments were carried out to study the effect of conditioning shift of the membrane potential on the mechanism of action-potential generation. The effect of conditioning was assessed from changes in the action-potential curve and its first derivative, as well as from the curve of transmembrane currents under voltage clamp conditions. It was found that a change in membrane potential evokes at least two reactions which have opposite effects on the mechanism of generation of action potentials. These reactions evidently have different time characteristics. One of these does not differ notably from the reaction recorded for other excitable structures, and is manifested in the activation (with hyperpolarization) or inactivation (with depolarization) of the mechanism generating action potentials. The other reaction contributes either to an increase (with depolarization) or a decrease (with hyperpolarization) in the efficiency of this mechanism. Conditioning polarization also has a marked effect on the system responsible for repolarization of the membrane during generation of action potentials. This effect is manifested in a change in the reaction of this system to tetraethylammonium ions. The specific membrane systems sustaining excitability and reacting to changes in the strength of the membrane's electrical field were found to be very inert. After a shift in the potential to a given stable level a rearrangement, lasting sometimes tens of seconds, takes place in the membrane.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 2, No. 1, pp. 91–99, January–February, 1970.     

Kinetics of enzymatic degradation of cyanide

CYANIDASE(@) is a new enzyme preparation capable of degrading cyanide in industrial wastewaters to ammonia and formate in an apparently one-step reaction, down to very low concentrations. This enzyme has both a high selectivity and affinity toward cyanide. A granular form of the biocatalyst was used in a recirculation fixed bed reactor in order to characterize the new biocatalyst with respect to pH, ionic strength, common ions normally present in wastewaters, mass transfer effects, and temperature. Long term stability was investigated. The kinetics of the enzymatic degradation of cyanide were studied in a batch reactor using the powdered immobilized enzyme preparation and modeled using a simple Michaelis-Menten equation.     

Effects of varying periods of immobilization of a limb on the morphology of a peripheral nerve

Immobilization of a hind limb of the cat in the resting position was found to result in morphological changes in the nerve supplying the medial head of the gastrocnemius muscle. A reduction in the diameter of the larger myelinated fibres (greater than 12 micrometers) concomitant with a reduction in the mean fibre diameter of the nerve were observed. This reduction in the mean fibre diameter of the nerve was found to be directly proportional to the period of immobilization. Prolonged immobilization beyond 8 weeks, resulted in the splitting of the myelin lamellae, formation of myelin rings and figures in some of the larger axons in addition to an abundant increase in collagen. These structural changes indicate a definite neural involvement in the disuse atrophy of the muscle as a result of immobilization extending for a period of 8 weeks and above.     

Computer-based screening of functional conformers of proteins

A long-standing goal in biology is to establish the link between function, structure, and dynamics of proteins. Considering that protein function at the molecular level is understood by the ability of proteins to bind to other molecules, the limited structural data of proteins in association with other bio-molecules represents a major hurdle to understanding protein function at the structural level. Recent reports show that protein function can be linked to protein structure and dynamics through network centrality analysis, suggesting that the structures of proteins bound to natural ligands may be inferred computationally. In the present work, a new method is described to discriminate protein conformations relevant to the specific recognition of a ligand. The method relies on a scoring system that matches critical residues with central residues in different structures of a given protein. Central residues are the most traversed residues with the same frequency in networks derived from protein structures. We tested our method in a set of 24 different proteins and more than 260,000 structures of these in the absence of a ligand or bound to it. To illustrate the usefulness of our method in the study of the structure/dynamics/function relationship of proteins, we analyzed mutants of the yeast TATA-binding protein with impaired DNA binding. Our results indicate that critical residues for an interaction are preferentially found as central residues of protein structures in complex with a ligand. Thus, our scoring system effectively distinguishes protein conformations relevant to the function of interest.     

The Significance of Non-vertical Transmission of Phenotype for the Evolution of Altruism

My aim in this paper is to demonstrate that a very simple learning rule based on imitation can help to sustain altruism as a culturally transmitted pattern or behaviour among agents playing a standard prisoner’s dilemma game. The point of this demonstration is not to prove that imitation is single-handedly responsible for existing levels of altruism (a thesis that is false), nor is the point to show that imitation is an important factor in explanations for the evolution of altruism (a thesis already prominent in the existing literature). The point is to show that imitation contributes to the evolution of altruism in a particular way that is not always fairly represented by evolutionary game theory models. Specifically, the paper uses a simple model to illustrate that cultural transmission includes mechanisms that do not transmit phenotype vertically (i.e. from parent to related offspring) and that these mechanisms can promote altruism in the absence of any direct biological propensity favouring such behaviour. This is a noteworthy result because it shows that evolutionary models can be built to explicitly reflect the contribution of non-vertical transmission in our explanations for the evolution of altruism among humans and other social species.     

Effects of immobilization of a limb on the maturation of a peripheral nerve in kittens

Right or left hindlimbs of kittens were immobilized in a plaster cast, in the resting position. Three groups of animals belonged to the same age group, one to older, and another to younger. Kittens in the same age group were subjected to progressively increasing periods of immobilization, the older group to a shorter and the younger to the longest period. By virtue of this arrangement, it was found that the demonstrated changes in the peripheral nerve, which included a decrease in the myelinated nerve fibre population and a reduction in the mean nerve fibre diameter, have been proportionate among the animals in the same age group, more pronounced in younger, and less so in older ones, indicating that in younger animals longer periods of immobilization lead to a greater degree of harmful effects, although it was also evident that younger animals attempt to adapt to such adverse conditions.     

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.     

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.