Purification of acetylcholinesterase from horse erythrocytes using affinity chromatography

A commercial preparation of water-soluble acetylcholinesterase from horse red cells has been purified to a specific activity of 2380 U/mg of protein (a 1660-fold purification) by a twofold affinity chromatography on the known sorbent of Sepharose-p-[NH-(CH2)5-C(O)NH(CH2)5C(O)NH-]-C6H4-N+(CH3)3 X Br- at pH 7.5. A selective elution of the enzyme was carried out from 10 mM of the phosphate buffer solution which contains 0.2% of triton X-100. Subsequent desorption of the enzyme proceeded with 5 mM of phenyltrimethylammonium bromide introduced into the buffer. Such effective preparations of acetylcholinesterase have not been previously produced. Effectiveness of the affinity sorbents considerably depends on the nature of the ligand which is covalent-linked with a Sepharose matrix and on the length of the attachment spacer arm ("insert") between them. A reversible inhibitory effect of certain ligands (tetramethylammonium, phenyltrimethylammonium) and their derivatives on acetylcholinesterase is estimated in comparison.     

Evolution of the genetic code: partial optimization of a random code for robustness to translation error in a rugged fitness landscape

Background  

The standard genetic code table has a distinctly non-random structure, with similar amino acids often encoded by codons series that differ by a single nucleotide substitution, typically, in the third or the first position of the codon. It has been repeatedly argued that this structure of the code results from selective optimization for robustness to translation errors such that translational misreading has the minimal adverse effect. Indeed, it has been shown in several studies that the standard code is more robust than a substantial majority of random codes. However, it remains unclear how much evolution the standard code underwent, what is the level of optimization, and what is the likely starting point.     

Generalized quasispecies model on finite metric spaces: isometry groups and spectral properties of evolutionary matrices

The quasispecies model introduced by Eigen in 1971 has close connections with the isometry group of the space of binary sequences relative to the Hamming distance metric. Generalizing this observation we introduce an abstract quasispecies model on a finite metric space X together with a group of isometries \(\Gamma \) acting transitively on X. We show that if the domain of the fitness function has a natural decomposition into the union of tG-orbits, G being a subgroup of \(\Gamma \), then the dominant eigenvalue of the evolutionary matrix satisfies an algebraic equation of degree at most \(t\cdot \mathrm{rk}_{\mathbf {Z}} R\), where R is the orbital ring that is defined in the text. The general theory is illustrated by three detailed examples. In the first two of them the space X is taken to be the metric space of vertices of a regular polytope with the natural “edge” metric, these are the cases of a regular m-gon and of a hyperoctahedron; the final example takes as X the quotient rings \(\mathbf {Z}/p^n\mathbf {Z}\) with p-adic metric.

     

Mathematical modeling of tumor therapy with oncolytic viruses: Regimes with complete tumor elimination within the framework of deterministic models

Background  

Oncolytic viruses that specifically target tumor cells are promising anti-cancer therapeutic agents. The interaction between an oncolytic virus and tumor cells is amenable to mathematical modeling using adaptations of techniques employed previously for modeling other types of virus-cell interaction.     

Dynamics of hematologic parameters and of the erythrocyte deformability index at the juvenal period of rats and guinea pigs

In experiments on Wistar rats and guinea pigs, dynamics of blood hematological parameters and of erythrocyte deformability index (DI) was studied from birth to the 3-month age. In rats, significant changes were revealed in the picture of blood for the first month of postnatal ontogenesis. The number of erythrocytes and leukocytes sharply increased, the number of reticulocytes decreased, their ratio in the degree of maturity was changed. The mean erythrocyte volume and their acidic resistance also decreased. Prior to the 4-week age, DI increased, which was accompanied by an increase in the hemoglobin hydration and flattening of the erythrocyte shape. Then DI decreased and erythrocytes acquired the more spherical shape. After the critical period of 3–4 weeks the studied parameters reached plateau. In guinea pigs, at this period of ontogenesis the hematologic parameters had sufficiently stable values, except for a DI decrease from birth to 1.5 months, then it rose without reaching the initial values. Like in the case with rats, DI had a close correlation with the erythrocyte parameter of shape (O_min).     

Dynamics of hematological blood parameters of white rats in postnatal ontogenesis

In experiments on Wistar rats, dynamics of blood hematologic and rheological parameters was studied from birth to the 3-month age. Results of this study indicate an intensive activity of the red bone marrow anlagen for the first 3 weeks of postnatal ontogenesis. This is manifested as an increase of the number of cells in the erythron system, a change of the erythrocyte acidic resistance, and a shift in the leukocytic formula. The boundary between the 2nd and 3rd weeks of ontogenesis is a critical period: there occur a sharp deceleration of erythropoiesis and a change of the age-related erythrocyte composition and of the blood leukocyte ratio towards the parameters characteristic of adult rats. The rise in the number of leukocytes and erythrocytes does not stop after this period. Until the 4-week age, the deformability index increases to be accompanied by an increase of hemoglobin hydration and improvement of the erythrocyte toroid shape. Subsequently, inversion of these processes occurs.     

Myxomycete biodiversity of the Colorado Plateau

A rapid biodiversity assessment for myxomycetes (plasmodial slime moulds) was carried out during a two-week field trip to the Colorado Plateau (western U.S.A.). Due to the very arid climate of the region, the moist chamber culture technique formed a major part of the survey. A total of 1165 records belonging to 93 species and 1 variety from 27 genera was collected from the 433 moist chamber cultures prepared with the bark surface of living plants, litter and weathered dung of herbivorous animals. Only 31 specimens of 16 wood-inhabiting species were collected in the field, mainly at higher elevations. Subsequent moist chamber cultures produced numerous bark-inhabiting species that are usually considered to be rare, including Echinostelium coelocephalum, Protophysarum phloiogenum, and Macbrideola declinata. The most common litter-inhabiting species were Didymium mexicanum and Badhamia melanospora (mainly on decaying parts of Opuntia spp. and Agave spp.); the most abundant coprophilous species were Badhamia cf. apiculospora, Fuligo cinerea, Perichaena liceoides, and Licea tenera. Both, species richness and diversity, increase from sagebrush desert to pine-juniper open woodland and pine-oak woodland. The myxomycete biota of the Colorado Plateau displays a high level of similarity to those of other arid regions of the world (mean coefficient of community C_s = 0.67), differing considerably from temperate (mean C_s = 0.34), boreal (mean C_s = 0.49) and tropical biotas (mean C_s = 0.42).     

Population models with singular equilibrium

A class of models of biological population and communities with a singular equilibrium at the origin is analyzed; it is shown that these models can possess a dynamical regime of deterministic extinction, which is crucially important from the biological standpoint. This regime corresponds to the presence of a family of homoclinics to the origin, so-called elliptic sector. The complete analysis of possible topological structures in a neighborhood of the origin, as well as asymptotics to orbits tending to this point, is given. An algorithmic approach to analyze system behavior with parameter changes is presented. The developed methods and algorithm are applied to existing mathematical models of biological systems. In particular, we analyze a model of anticancer treatment with oncolytic viruses, a parasite-host interaction model, and a model of Chagas' disease.     

Changes of physiological and biochemical characteristics of rat erythrocytes after blood loss

In experiments of Wistar male rats, changes are studied of erythrocyte hematological, biochemical (activitities of transport ATPases), and rheological properties (capability for aggregation and deformability) 7 days after bloodletting of 12-15 % of the total blood mass. It has been shown that alongside with an elevation of erythrocyte volume and of the number of immature cells - reticulocytes, there was a statistically significant increase of Na,K-ATPase and Ca-ATPase activities in the whole erythrocytes and their membrane preparations - ghosts, the increment of activity in the case of Na,K-ATPase being essentially higher in the whole cells. This indicates the appearance of an enzyme activator inside the erythrocytes. There are also revealed a decrease of firmness of erythrocyte aggregates, a deceleration of spontaneous aggregation, and an increase of index of erythrocyte deformability. The conclusion is made that changes of erythrocyte rheological properties are interconnected with changes of the Na,K-ATPase activity and are directed to optimization of blood circulation in large vessels and capillary network.