Structural and functional analysis of the respiratory burst in the phagocytosing macrophage

The interrelation between structural changes and oxygen consumption by the phagocyting macrophage was studied. The mean number of phagocyted particles was estimated by the method of stereological transformation. It is found that the uptake of yeast particles and CN- -nonsensitive oxygen consumption is related to the concentration of yeast cells in the incubation medium. A positive correlation was established between the oxygen consumption and the mean number of phagocyted particles. The results obtained may suggest that the "respiration burst" takes place in the contact area of the macrophage and the phagocyted material, and its extent probably depends on the surface of that contact area.     

Positive selection is a general phenomenon in the evolution of abalone sperm lysin

Lysin is a 16kDa acrosomal protein used by abalone sperm to create a hole in the egg vitelline envelope (VE). The interaction of lysin with the VE is species-selective and is one step in the multistep fertilization process that restricts heterospecific (cross-species) fertilization. For this reason, the evolution of lysin could play a role in establishing prezygotic reproductive isolation between species. Previously, we sequenced sperm lysin cDNAs from seven California abalone species and showed that positive Darwinian selection promotes their divergence. In this paper an additional 13 lysin sequences are presented representing species from Japan, Taiwan, Australia, New Zealand, South Africa, and Europe. The total of 20 sequences represents the most extensive analysis of a fertilization protein to date. The phylogenetic analysis divides the sequences into two major clades, one composed of species from the northern Pacific (California and Japan) and the other composed of species from other parts of the world. Analysis of nucleotide substitution demonstrates that positive selection is a general process in the evolution of this fertilization protein. Analysis of nucleotide and codon usage bias shows that neither parameter can account for the robust data supporting positive selection. The selection pressure responsible for the positive selection on lysin remains unknown.      

Galactomannan from Ambiguous Crazyweed (Oxytropis ambigua(Pall.) DC)

A galactomannan with a molecular weight of 735 kDa was first isolated and purified from seeds of ambiguous crazyweed Oxytropis ambigua(Pall) DC (family Leguminosae) with a yield of 3.6%. Its aqueous solutions displayed an optical activity ([] _D = 73.32°) and high viscosity ([] = 644 ml/g). Chemical analysis and ¹³C-NMR spectroscopy revealed the presence of D-mannopyranose and D-glucopyranose in the heteropolysaccharide at a molar ratio of 1.39 : 1. The linear backbone of its macromolecule consists of 1,4--D-mannopyranose residues. Single -D-galactose residues substitute 72% of the mannoses to form branches.     

Solid phase synthesis of diamides as potential bone resorption inhibitors

Unsymmetrical diamide libraries have been prepared by a general and versatile solid phase route, using diacid templates in combination with aromatic and aliphatic amines chosen with the help of statistical experimental design. The compounds were tested as potential inhibitors of osteoclast vacuolar ATPase.     

Study of galactoglucomannans from the seed of Cercis canadensis L

Galactoglucomannans were isolated by selective precipitation from aqueous and alkaline extracts of endosperm and hulls of Cercis canadensis, a member of the family Fabaceae. Their monosaccharide composition (Man: Gal: Glu) was as follows: 10.4: 0.9: 1 (polysaccharide from the endosperm) and 4.5: 0.9: 1 (polysaccharide from the hulls). The identity of IR spectra was indicative of the commonness of their structure. Analysis of the structure of the galactoglucomannan from endosperm by 13C NMR spectroscopy showed that its main chain consisted of 1,4-beta-D-manno- and 1,4-beta-D-glucopyranose. Part of mannose residues in the chain were substituted at C6 with single residues of alpha-D-galactopyranose. Galactoglucomannans are located in different part of the seed and implement different functions.     

Study of galactoglucomannans from the seed of <Emphasis Type="Italic">Cercis canadensis</Emphasis> L.

Galactoglucomannans were isolated by selective precipitation from aqueous and alkaline extracts of endosperm and hulls of Cercis canadensis, a member of the family Fabaceae. Their monosaccharide composition (Man : Gal : Glu) was as follows: 10.4 : 0.9 : 1 (polysaccharide from the endosperm) and 4.5 : 0.9 : 1 (polysaccharide from the hulls). The identity of IR spectra was indicative of the similarity of their structure. Analysis of the structure of the galactoglucomannan from endosperm by ¹³C NMR spectroscopy showed that its main chain consisted of 1,4--D-manno- and 1,4--D-glucopyranose. Part of the mannose residues in the chain were substituted at C6 with single residues of -D-galactopyranose. Galactoglucomannans are located in different parts of the seed and implement different functions.__________Translated from Prikladnaya Biokhimiya i Mikrobiologiya, Vol. 41, No. 3, 2005, pp. 324–329.Original Russian Text Copyright © 2005 by Mestechkina, Egorov, Anulov, Shcherbukhin.     

Redistribution of subpopulations of peritoneal macrophages with Fc gamma receptors under the effect of an immunomodulator

The heterogeneity of resident peritoneal macrophages and peritoneal macrophages was studied in different periods following oral administration of sodium nucleinate according to their ability to bind and phagocytize sheep erythrocytes opsonized by means of specific rabbit IgG. Using a mathematical method developed earlier, it has been possible to demonstrate that resident peritoneal macrophages can be divided into two subpopulations--actively and poorly binding macrophages but, after activation by sodium nucleinate--into three subpopulations. Fractionation of the macrophages according to their ability to adhesion within a temperature gradient has shown that the same peaks are traced in the fractions as in the overall pool of cells, but in different quantitative ratios. It has also been demonstrated that phagocytic activity is reduced in macrophages capable of adhesion to plastic at lower temperatures.     

Anticoagulant activity of low-molecular-weight sulfated derivatives of galactomannan from Cyamopsis tetragonoloba (L.) seeds

Galactomannan from seeds of Cyamopsis tetragonoloba (L.) Taub. (guar) was depolymerized using immobilized enzymatic preparation celloviridin. A set of fragments whose molecular weights varied from 12.6 to 245.6 kDa was obtained. Sulfated derivatives of components of all fractions were synthesized, in which the content of HSO3(-) groups was 48.05% +/- 2.31. All preparations exhibited anticoagulant activity, which was recorded in vitro in two tests--aIIa and aXa. The antithrombin activity (aIIa) was high (up to 65-87 U/mg) and did not depend on the molecular weight of a sulfated derivative; in the second test (aXa), the effect of molecular weight was observed. Biospecific electrophoresis allowed us to detect the ability of galactomannan sulfates to form complexes with protamine sulfate, a classic antidote to heparin.     

Determination of the Primary and Fine Structures of a Galactomannan from the Seed of Gleditsia triacanthos f. inermis L.

Galactomannan, a polysaccharide with a molecular weight of 660 kDa, was isolated for the first time from the seed of Gleditsia triacanthos f. inermis (yield, 15.4%). Its aqueous solutions were optically active ([] _D = +31.0°) and highly viscous ([] = 578 ml/g). Analysis of this heteropolysaccharide using chemical, enzymatic, and chromatographic procedures, as well as IR and ¹³C NMR spectroscopy, showed that it consists of D-mannopyranose and D-galactopyranose residues (molar ratio, 2.42 : 1). The main chain of this galactomannan comprises 1,4--D-mannopyranose residues, 41% of which are substituted at C6 with single residues of -D-galactopyranose. The probability of occurrence in the chain of mannobiose units substituted otherwise, determined experimentally, was 0.16 for the Man–Man unit, 0.50 for the Gal(Man–Man) and (Man–Man)Gal units, and 0.34 for the disubstituted Gal(Man–Man)Gal unit.     

Mitochondrial DNA and bindin gene sequence evolution among allopatric species of the sea urchin genus Arbacia

Sea urchins of the genus Arbacia (order Stirodonta) have discontinuous allopatric distributions ranging over thousands of kilometers. Mitochondrial DNA (mtDNA) sequences were used to reconstruct phylogenetic relationships of four Arbacia species and their geographic populations. There is little evidence of genetic structuring of populations within species, except in two cases at range extremes. The mtDNA sequence differentiation between species suggests that divergence occurred about 4-9 MYA. Gene sequences encoding the sperm protein bindin and its intron were obtained and compared with the mtDNA phylogeny. Sea urchins among the well-studied echinoid order Camarodonta, with degrees of mtDNA divergence similar to those of Arbacia species, are known to have remarkable variation in bindin. However, in Arbacia, little variation in deduced amino acid sequences of bindin was found, indicating that purifying selection acts on the protein. In contrast, bindin intron sequences showed much differentiation, including numerous insertion/deletions. Fertilization experiments performed between a divergent pair of Arbacia species from the Atlantic and Pacific Oceans revealed no evidence of blocks to gamete recognition. In Arbacia, fertilization specificities may have evolved relatively slowly as a result of extensive gene flow within species, greater functional constraint on the bindin polypeptide, or reduced selective pressure for species recognition in singly occurring species.