Purification of neuraminidase from influenza viruses by affinity chromatography. The neuraminidase (acylneuraminyl hydrolase, EC 3.2.1.18) of the influenza virus recombinant strain (HON2) was solubilized with detergents and isolated by affinity chromatography. The neuraminidase could be purified to a single high molecular weight glycoprotein when assayed under non-reducing conditions on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme showed an increase in specific activity from 2.46 to 189 micronM N-acetylneuraminic acid released per min per mg protein and the recovery represented 123% of the activity of intact virus particles. The enzyme could be purified from laboratory preparations of virus or from outdated influenza virus vaccine. Viral neuraminidases purified by this technique were stable at pH 6.0 for several hours.


The interaction of p-nitrophenyl carbamate with urease. 1. p-Nitrophenyl carbamate and thiourea have been shown to be substrates for urease (urea amidohydrolase, EC 3.5.1.5) 2. Urease has been shown to have a lower Km, 0.67 mM, with p-nitrophenyl carbamate than with urea, 2.0 mM. 3. The V of urease for the hydrolysis of urea, p-nitrophenyl carbamate and thiourea has been shown to be the same, indicating a common rate-limiting step. 4. A mechanism has been proposed for urease where the initial rate-limiting step is the release of a molecule of ammonia from the substrates.


Mechanistic studies on the biosynthesis of 6-ketoprostaglandin F1alpha. The isolation, structure and biosynthesis of a novel metabolite of arachidonic acid formed during incubations of rat stomach homogenates is reported. This product was formed from unlabeled, tritium-labeled and deuterium-labeled arachidonic acid and the prostaglandin endoperoxides, G2 and H2, demonstrating its formation via the "prostaglandin endoperoxide synthetase" pathway. Its structure, 6-ketoprostaglandin F1alpha, was based on mass spectral interpretation of several derivatives (undeuterated and deuterated) and confirmed through chemical synthesis. 6-Ketoprostaglandin F1alpha is unique in being formed from a substrate of the "2" series of prostaglandins, i.e. arachidonic acid, yet belonging to the "1" series. A mechanism for its biosynthesis is proposed involving a specific cyclising enzyme which we term, 6(9)-oxycyclase.


The role of diacylglycerol-carrying lipoprotein I in lipid transport during insect vitellogenesis. A diacylglycerol-carrying lipoprotein was isolated from mature eggs of the silkworm, Philosamia cynthia and compared, for physiochemical properties, with the major diacylglycerol-carrying lipoprotein I (LP-I) of hemolymph. The two molecules are identical an electrophoretic mobility, structural configuration as revealed by electron microscopy, and amino acid composition. In addition mannose was detected in the lipid-free protein moiety, thus enabling classification of the molecules as glycoproteins. The molecules differ in lipid content with egg-LP-I containing only 3.6% of the diacylglycerol content of hemolymph LP-I and the phospholipid and cholesterol components also showing a marked reduction. Analysis of the LP-I and vitellogenin (another diacylglycerol-carrying lipoprotein; LP-II) concentrations of mature eggs indicates that the amounts of the two glycolipoproteins in eggs are insufficient to account for the total acylglycerol content of the eggs. The results suggest that LP-I functions as a true carrier-protein serving to transport diacylglycerol from the fat body to the ovary. This proposal is supported by the observation that LP-I isolated from the egg retains the physiological capacity of hemolymph-LPi to take up diacylglycerol from fat body. Thus it is suggested that LP-I is the major source of lipid for vitellogenesis, whereas vitellogenin is the primary source of protein.


The primary structure of phospholipase A2 from porcine pancreas. A reinvestigation. The primary structure of porcine pancreatic phospholipase A2 (EC 3.1.1.4) has been reinvestigated. A number of modifications have been introduced including the addition of a 7th disulfide bridge. The structure which is presented here shows a high degree of homology with the amino acid sequence of snake venom and horse pancreas phospholipase A2.


[Determination of the primary structure of alfalfa mosaic virus (strain S) coat protein. II. Complete sequence of the protein (author's transl)]. The alfalfa mosaic virus protein was submitted to the action of cyanogen bromide. Four peptides were isolated. Study of these peptides allowed us to determine the order. Then protein was submitted, after S-carboxymethylation or S-aminoethylation, to the action of different proteolytic enzymes: trypsin, chymotrypsin, thermolysin and papain. The peptides issued from these different hydrolysis were separated on Dowex 50 X4 and Dowex 1 X2, and their amino acid composition was determined. The use of classical methods of sequence determination, of mass spectrometry and for one case the use of a sequencer, lead to the obtention of the primary structure of all the tryptic peptides. The studies of chymotryptic, thermolytic and papainic hydrolysates, and of cyanogen bromide rupture, allowed us to isolate the overlapping peptides which were necessary for the reconstitution of the complete proteic chain.


Properties of carbohydrate-stripped thyroglobulin. III. Solubility characteristics of thyroglobulin. In studying the solubility characteristics of thyroglobulin, it was found that the removal of sialic acid residues from either human or hog thyroglobulin by treatment with neuraminidase markedly decreased the solubility in salt solutions. On the other hand, naturally sialic acid-rich thyroglobulin obtained from human diffuse goiter was slightly more soluble than thyroglobulin in which sialic acid content was within the normal range. However, the desialized preparations prepared from both normal and sialic acid-rich thyroglobulin showed markedly lower solubility and further, showed no significant difference in solubility between them. These results indicate that the surface charge of thyroglobulin which influenced the salting-out property depends largely on the number of sialic acid residues in a thyroglobulin molecule, not on the degree of iodination of thyroglobulin.


Heterogeneity of purified cholera toxin. The heterogeneity of Vibrio cholerae toxin, obtained from culture filtrates in homogeneous form by gel filtration and preparative disc gel electrophoresis has been studied. By means of disc electrophoresis on polyacrylamide gel cholera toxin was separated into three forms designated I (5%), II (15%) and III (80%). The toxic activity, amino acid content and molecular weight of the three forms were similar. The difference so far observed between the various electrophoretic fractions is a difference in net charge. Incubation of either cholera toxin II or cholera toxin III at relatively high pH leads to the formation of the more acidic forms. These forms, generated in vitro by deamidation of asparagine and/or glutamine residues, are indistinguishable from the toxins of similar electrophoretic mobilities isolated from crude culture filtrates.


Reactions of gold(III) ions with ribonuclease A and methionine derivatives in aqueous solution. 1. Addition of Au(III) (as chloroaurate) to folded ribonuclease A causes the formation of aggregates, as shown by chromatography and extreme broadening of the 270 MHZ 1H NMR spectrum. 2. When RNAase is partially unfolded Au(III) causes oxidation of methionines to the sulphoxide, and leads to almost complete unfolding (at molar equivalent ratios). 3. Reactions between model methionine derivatives and Au(III) show that the oxidation involves the production of Au(I)-methionine species. The stability of these complexes is dependent on the availability of free NH2 groups which catalyse their disproportionation.


Studies of disulfide bond location in ovine lutropin beta subunit. By combinations of selective chemical cleavage (cyanogen bromide), selective enzymatic cleavage (trypsin, thermolysin), and random cleavage (partial acid hydrolysis) a series of disulfide-containing peptides have been isolated from ovine lutropin beta subunit. These peptides suggest six disulfide linkages between half-cystine residues in positions 23-72, 26-110, 93-100, 34-88, 9-90, and 38-57. The latter pair was placed by elimination of other possibilities. The first three pairs are in agreement with a report by Chung, D., Sairam, M. R. and Li, C. H. (1975) Int. J. Peptide Protein Res. 7, 487-493; the pair 93-100 has also been detected by Reeve, J. R., Cheng, K. W. and Pierce, J. G. (1975) Biochem. Biophys. Res. Commun. 67, 149-155, using partial reduction and alkylation. In an attempt to improve the efficiency of enzymatic attack, a preliminary partial reduction as per Reeve et al. [16] was done. In this instance a peptide suggesting an additional disulfide linkage between half-cystines 23-26 was obtained as well as peptides consistent with the 23-72 and 26-110 placements. This was interpreted as an artifactual opening and recombining during partial reduction-reoxidation to produce the 23-26 linkage. The placement of three disulfide bonds (34-88, 9-90, and 38-57) is in disagreement with the pairings Chung et al. [15] suggest for these six half-cystine residues. Six reasons for uncertainty in the placement of disulfide bonds are discussed. It is concluded the definitive placement of the disputed three disulfide bonds in ovine lutropin beta subunit remains an open question.


Electronmicroscopic investigation of the flexibility of F-actin. The contour lenghts and the end-to-end distances of a large number of F-actin filaments were measured in electronmicrographs. Preparation of F-actin for electron-microscopy was made at three different temperatures. The flexibility parameter or the elastic modulus for bending of F-actin was determined from the relation between the contour length and the end-to-end distance after statistical treatment. The value of the flexibility parameter obtained here showed good agreement with that obtained by the quasielastic light scattering at all temperatures examined. The flexibility of F-actin increased with raising temperature and with the binding of heavy meromyosin.


Isolation and characterization of a low-molecular weight seed protein from Ricinus communis. A seed protein with an apparent molecular weight of 5000 has been isolated and purified from Ricinus communis. This protein has a very high content of glutamine and an unusual ultraviolet absorption spectrum. The amino-terminal sequence of 22 residues determined by automatic Edman degradation is: NH2-Pro-Ser-Gln-Gln-Gly-Cys-Cys-Gly-Gln-Ile-Gln-Glu-Gln-Gln-Asn-Leu-Arg-Gln-Cys-Gln-Glu-Tyr-.


Conformational transitions of alpha-elastin. The alpha-elastin, the soluble product obtained from elastin by oxalic acid hydrolysis, has been studied from a conformational point of view in different conditions using circular dichroism spectroscopy. The spectral variations of the alpha-elastin dissolved in water and in different organic solvents/water mixtures have been examined in order to establish the effect of the solvents on the structural properties of the protein. The effect of calcium ions has also been studied on alpha-elastin dissolved either in water or in organic solvents. Finally, the effect of pH and temperature on the conformational properties of the alpha-elastin has been investigated.


Some observations on the ageing in vitro of reprecipitated collagen fibres. The changes in solubility and amounts of reducible cross-links have been studied during "ageing" in vitro of reprecipitated rat skin collagen fibres by incubation at 37 degrees C. Fibres from pre-reduced collagen devoid of aldehyde precursors became insoluble at the same rate as that of normal fibres during "ageing". Insolubilization occurred at a much faster rate in the presence of oxygen than in air and was almost completely inhibited when oxygen was excluded. The rate of decline of the reducible cross-links was, however, unaffected by oxygen tension. The results indicate that, during "ageing" in vitro, conversion of the lysine-derived cross-links to a non-reducible form is not associated with solubility changes. The relationship of these in vitro changes to those ocurring in vivo is unknown.


Structural and functional studies of hemoglobin J Cala-bria: beta64 (E8) Gly leads to Asp. A hemoglobin of high electrophoretic mobility was detected in a French male suffering from an acute leukemia; this hemoglobin was also present in his family. The variant was unstable and possessed an abnormal beta chain, in which a glycyl residue in position 64 (E8) was replaced by an aspartyl residue. This variant constitutes a new case of Hb J Calabria. Since the substituted E8 residue is in close spatial contact with that at B6, it was of interest to compare the properties of Hb J Calabria with those of other hemoglobins bearing substitutions at the same site.


Comparative effects of native insulin and insulin-dextran complexes on the metabolism of adipose tissue. Insulin-dextran complexes have been prepared and their biological activities compared to that of native insulin. Complexes referred to as T-70 had a molecular weight of about 450 000, T-2000 being more than 2 million. Insulin-dextran T-70 was stable and no release of free insulin from the complex was observed upon its incubation with adipose tissue. Insulin-dextran T-70 mimicked the effects of native insulin upon adipose tissue metabolism. It also lowered blood sugar. These effects necessitated, on an insulin molar basis, concentrations of the complex that were 10-times greater than those needed for native insulin. Maximal concentrations of insulin complex T-70 or native insulin elicited similar quantitative effects. This suggested that when the concentration of insulin-dextran T-70 was high enough, the complex occupied a sufficient number of receptor sites to produce maximal stimulation of the tissue. In contrast, insulin-dextran T-2000 was barely effective, indicating that, probably due to its size, it was unable to reach receptor sites. The size, stability and metabolic effects of insulin-dextran T-70 observed in this study give additional support to the concept that insulin action is probably mediated via a series of events initiated at the level of the plasma membrane of adipocytes.


Isolation and characterization of the sulfated glycosaminoglycans of the vitreous body. Ester sulfate containing glycosaminoglycans comprising approx. 3% of the total glycosaminoglycan content, have been isolated from protease-digested bovine vitreous body by stepwise fractionation on AG-1X2(Cl-) and gel filtration on Bio-Gel P-300. Two heparan sulfate and two chondroitin-4-sulfate fractions were isolated in nearly pure form. The heparan sulfate fractions were undersulfated and contained the same relative proportions of N- and O-sulfate (1:2), although the total sulfate content differed by approx. 100%. No chondroitin-6-sulfate was present in the isolates, based on evidence obtained from chondroitin ABC lyase experiments.


Dissociation of aggregated ferroheme complexes and protoporphyrin IX by water-soluble polymers. The ferroheme-pyridine complex, ferroheme and protoporphyrin IX form the aggregates by the hydrophobic interaction in aqueous solutions. We found by spectrophotometric and fluorometric measurements that the aggregates dissociated into the monomers by the addition of water-soluble polymers, such as, poly(ethyleneoxide), poly(vinylalcohol), poly(vinylpyrrolidone) and poly(styrene sulfonate). The dissociation by the polymers proceeded as their hydrophobicities increased. The aggregated ferroheme was effectively dissociated by the copolymers of 4-vinylpyridine which were water-soluble polymer-ligands.


Studies on bacterial cell wall inhibitors. III. 3-amino-3-deoxy-D-glucose, an inhibitor of bacterial cell wall synthesis. It was shown that 3-amino-3-deoxy-D-glucose, one of the constituents of the kanamycin molecule and a metabolite of Bacillus sp., inhibits the bacterial synthesis of cell wall. The antibiotic (100 microgram/ml) significantly inhibits the growth of Staphylococcus aureus FDA 209P as well as the incorporation of DL-[14C]alanine into the acid-insoluble macromolecular fraction of its growing cells in the presence of chloramphenicol (100 microgram/ml). In contrast, the antibiotic does not affect the incorporation of [3H]thymidine, [3H]uridine and L-[14C]leucine. The other constituents of kanamycin, 6-amino-6-deoxy-D-glucose and deoxystreptamine do not inhibit the synthesis of bacterial cell wall peptidoglycan.


Obligatory free radical intermediate in the oxidative activation of the carcinogen N-hydroxy-2-acetylaminofluorene. We have demonstrated that the nitroxyl free radical form of the carcinogen N-hydroxy-2-acetylaminofluorene (OH-AAF) is an obligatory intermediate in the cumene hydroperoxide-hematin-induced oxidative activation of this carcinogen into 2-nitrosofluorene and N-acetoxy-2-acetylaminofluorene. Both the rate of N-OH-acetylaminofluorene oxidation and the amount of its nitroxyl free radical were experimentally observed as a function of reaction time. Rate equations were derived for a model in which the nitroxyl free radical form of OH-AAF was an obligatory intermediate in the reaction. Using this theory it was possible to compute one experimental variable, the rate of OH-AAF oxidation, utilizing the other experimental variable, the amount of nitroxyl free radical present at any time during the reaction. The theory also predicts a linear relationship between the rate of OH-AAF oxidation and the square of the free radical content; and this was found to be true experimentally. The dismutation rate constant of the nitroxyl free radical of OH-AAF was found to be 2.7-10(5) M-1-s-1.


Release of O-sulfate groups under mild acid hydrolysis conditions used for estimation of N-sulfate content. The treatment of chondroitin sulfate isolated from cultured B16 mouse melanoma cells with 0.04 M HCl at 100 degrees C for 90 min released up to 45% of O-sulfate residues as free inorganic sulfate. In addition to the release of inorganic sulfate, extensive degradation of this polysaccharide as well as of cartilage chondroitin sulfate, pig rib cartilage proteoglycan, heparin and hyaluronic acid was also evident under these conditions. The above hydrolysis conditions are used for characterizing 35S-labeled heparan sulfates synthesized by cultured cells and to calculate ratio of N- and O-sulfates in these molecules. Our results suggest that caution is necessary in interpreting the results of mild acid hydrolysis of glycosaminoglycans.


Melittin interactions with adenylate cyclase. Melittin, a basic polypeptide from bee venom, inhibits basal and thyrotropin-stimulated adenylate cyclase of beef thyroid membranes with a Ki approximately 10 micron. Although this property resides in the basic C-terminal and not the N-terminal portion of the molecule, inhibition is due primarily to its detergent-like nature rather than charge effects. There is also a small enhancing effect of both basal and thyrotropin-stimulated adenylate cyclase of 0.3-3 micron melittin.


The submicrosomal distribution in rat and boar testis of some enzymes involved in androgen and 16-androstene biosynthesis. The submicrosomal localization of enzymes involved in androgen and 16-androstene biosynthesis in rat and boar testis tissue has been studied. 16-androstene production was not evident in the rat testis but two enzymes concerned with androgen generation (17 alpha-hydroxy C21 steroid C-17,20 lyase; 17 beta-hydroxysteroid dehydrogenase, were found predominantly in agranular microsomes. In the boar testis the C-17,10 lyase had a similar distribution to the rat enzyme but the 17 beta-hydroxysteroid dehydrogenase was found to be evenly distributed between the two microsomal types. The enzyme system, "andien-betha synthetase", involved in the conversion of pregnenolone to 5,16-androstadien-3 beta-ol, was found mainly (66%) in the agranular microsomes but had a lower specific activity than those of the two enzymes of androgen biosynthesis that have been studied.


Synthesis in vitro of glycoprotein in regenerating rat liver. The metabolism of glucosamine in regenerating rat liver was studied in liver slices. [1-14C] Glucosamine was incorporated into acid-soluble fraction, rapidly converted to UDP-N-acetylhexosamine and transferred to acid-insoluble fraction. Electrophoretic analysis revealed that most of the radioactive macromolecules released from the slices to the incubation medium were plasma glycoproteins. The incorporation of [1-14C] glucosamine into UDP-N-acetylhexosamine significantly increased from 6 h to 48 h after partial hepatectomy. On the contrary, the incorporation into acid-insoluble fractions of slice and medium decreased to about 50% of the control values. The rate of transfer of N-acetylhexosamine from UDP-N-acetylhexosamine to acid-soluble fractions also decreased at 12 h and 48 h respectively. This indicates that the transfer of N-acetylhexosamine to glycoproteins decreases during 48 h of liver regeneration. The enhancement of [1-14C] glucosamine incorporation into UDP-N-acetylhexosamine is due to an accumulation of the label in the larger pool of this compound. Evidently, some control mechanism may operate on the transfer of N-acetylhexosamine to glycoproteins in regenerating rat liver.


Studies of nicotinic acetylcholine receptor protein from rat brain. II. Partial purification. The pharmacological specificity of the binding of 125I-labeled alpha-bungarotoxin to a 1% Emulphogene BC-720 extract of a rat brain particulate fraction has been investigated. The extract contains a component which possesses the binding characteristics of a nicotinic acetylcholine receptor protein. The crude soluble acetylcholine receptor protein was purified by affinity chromatography utilizing the alpha-neurotoxin of Naja naja siamensis as ligand and 1.0 M carbamylcholine chloride as eluant. A single, batch-wise, affinity chromatography procedure yields an average purification of 510-fold. When this purified material is treated a second time by affinity chromatography, purification as high as 12600-fold has been obtained. Binding of 125I-labeled alpha-bungarotoxin to this purified acetylcholine receptor protein is saturable with a Kd of 1 - 10(-8) M. Nicotine and acetylcholine iodide at concentrations of 10(-5) M inhibit 125I-labeled toxin-acetylcholine receptor protein complex formation by 41 and 61% respectively. At 10(-4) M, carbamylcholine chloride and (+)-tubocurarine chloride give respectively 52 and 82% inhibition. Eserine sulfate and atropine sulfate have no effect on complex formation at a concentration of 10(-4) M. These data support the isolation of a partially purified nicotinic acetylcholine receptor protein.


The micro-assay of the disaccharide isomers of chondroitin sulphate. Connective tissue extracts containing the 4- and 6-sulphated isomers of chondroitin sulphate can be measured at the microgram level (approximately 10 microgram) using colorimetric assays. The chondroitin sulphates are depolymerised to disaccharides using chondroitin ABC lyase (EC 4.2.2.4). The 4- and 6-sulphated disaccharides after treatment with acid periodate are determined using a thiobarbituric acid procedure. An alkaline borate p-dimethylaminobenzaldehyde reaction is used to measure 6-sulphated disaccharide, the 4-sulphated disaccharide not forming a chromogen. Unsulphated disaccharide causes interference in both assays and if present is separated prior to assay using paper electrophoresis fractionation.


High-pressure liquid chromatography and field desorption mass spectrometry of heme alpha, heme alpha dimethyl ester and acetyl heme alpha dimethyl ester. Field desorption mass spectrometry was shown to be a valid technique for determining the molecular weights of hemins and hemin esters, as well as of porphyrins. The observed base peaks of ligand-free protoheme IX, protoheme IX dimethyl ester, and protoporphyrin IX dimethyl ester correspond well to the molecular weights of these compounds and the base peak for hematoporphyrin corresponds to the molecular weight of this porphyrin minus two molecules of water. The technique was employed to confirm the molecular weights of heme alpha, heme alpha dimethyl ester, and acetyl heme alpha dimethyl ester. Heme alpha dimethyl ester was prepared by reaction of heme alpha with trimethyloxonium tetrafluoroborate and purified by high-pressure liquid chromatography. The isolated product was converted to the acetylated derivative by reaction with acetic anhydride and was subsequently purified by high-pressure liquid chromatography. A field desorption spectrum of heme alpha shows a base peak at 582 which is in agreement with previous deductions of the structure of this prosthetic group. Base peaks of the heme alpha ester and its acetylated derivative demonstrate that the two carboxyl groups have been methylated and the single hydroxyl group has been acetylated without further alteration of the molecule.


Axial pertubations on the electronic and magnetic properties of ferric porphyrins. II. Solvent effects on the proton NMR spectra of low-spin cyano complexes. The proton NMR spectra of a series of low-spin bis-cyano ferric complexes of tetraarylporphyrins and octaethylporphyrin in a variety of solvents have been recorded and analyzed. The hyperfine shifts are shown to be very sensitive to the solvent, experiencing an overall downfield bias as the solvent hydrogen-bonding donor strength increased. The characteristic pattern of the contact and dipolar shifts for the meso-aryl group in tetraarylporphyrin complexes are shown to permit a quantitative separation of the dipolar and contact contributions to the hyperfine shift. The separated components indicate that increased solvent hydrogen bonding strength significantly decreases the magnetic anisotropy of the iron and diminishes porphyrin replaced by iron pi bonding. The changes in anisotropy with solvent are shown to be consistent with the coordinated cyanide acting as a proton acceptor. Although similar solvent effects are found to be absent in bis-imidazole complexes, a downfield bias of half the magnitude of the bis-cyano complexes is observed in mixed cyano/imidazole complexes. Hence, the heme hyperfine shifts in cyano-metmyoglobins and -hemoglobins may serve as probes for the protonation of the distal histidyl imidazole.


Different action of hormonal stimulation on the biosynthesis of three pancreatic enzymes. The rates of biosynthesis of amylase, lipase and chymotrypsinogen were followed in rat pancreas in vivo after intravenous injection of the hormone cholecystokinin-pancreozymin (8 IDU/kg) associated with secretin (5 CU/kg). This pancreatic stimulation resulted in non-parallel variations of the rates of biosynthesis of the three studied enzymes, suggesting independent regulation of synthesis. The result of one stimulation was calculated in terms of quantities of enzymes synthesized by the pancreas in comparison to control. It was found that chymotrypsinogen, amylase and lipase productions were increased by 63, 26 and 10%, respectively, indicating that repeated cholecystokinin-pancreozymin plus secretin stimulations could induce "adaptation-like" modifications of the pancreatic enzyme content.


Oxygen binding to cobalt(II) proto-, deutero- and meso-porphyrin IX dimethyl ester complexes in organic solvents. The binding of oxygen to cobalt(II) meso, deutero- and proto-porphyrin IX dimethyl esters complexed with pyridine or 2-methylimidazole was investigated at -10 degress - -60 degrees C in toluene or DMF solution, and the thermodynamic data related with the binding were presented. The oxygen affinity of cobalt meso-porphyrin complex was larger by the factor of 2.0-1.4 than those of the other complexes where oxygen affinities were not explained by a simple electron-withdrawing capability of 2,4-substituents of the porphyrin ring. The oxygen binding property was, generally, dependent on the solvent, suggesting that the solvation affects appreciably the oxygen binding to the complexes. The oxygen affinities of cobalt porphyrin complexes in various organic solvents were compared with those of their apomyoglobin complexes. The differences of oxygen affinities between both systems decreased with increasing the size of 2,4-substituents, and it was in the following order on 2,4-substituted porphyrins: Deutero greater than Proto greater than Meso. It was suggested that the 2,4-substitutent effect on the oxygen affinity of cobalt myoglobin complexes was not only caused by the direct electronic effect on the central cobalt atom, but also controlled by the stereochemical interaction between apomyoglobin and the porphyrin.


Interactions of platinum complexes, peptides, methionine and dehydrogenases. Eight platinum-methionine complexes have been investigated as inhibitors of the alcohol (EC 1.1.1.1) and lactate (EC 1.1.1.27) dehydrogenase systems. While only one complex, Pt(Met)Cl2, had two halogen ligands on the platinum, all the complexes were able to inhibit both enzyme systems. Of the various methionine and histidine containing peptides evaluated, the dipeptide Met-Met gave the best protection against inhibition by N-alkyl-substituted ethylenediamine-platinum complexes. The histidine containing peptides gave a slight protection against the inhibition by beta-[Pt(Met)(NH3)Cl]Cl. Thus it appears that in the enzyme systems studied, the methionine acts as a strong ligand for platinum.


Kinetics of the substitution reaction of thiocyanate with tetrakis (3-N-methylphyridyl)porphinecobalt(III). The equilibrium constant for the first thiocyanate addition to Co(III)-tetra(3-N-methylpyridyl)porphine is a factor of two smaller than for the less basic 4-N-methylporphine isomer. This effect is primarily due to a larger Co-SCN dissociation rate constant for the 3-isomer.


Perfusion of intact skin by a saline solution of bis(glycinato) copper(II). The perfusion of intact cat skin by a saline solution of bis(glycinato) copper(II), labelled with Cu64, has been studied in a diffusion cell. It has been shown by counting the activity that the copper(II) complex perfuses the skin and over a period of 6-7 hr reaches the isotonic saline solution underneath. After this inaugural delay period a steady rate of penetration was maintained and after 24 hr about 1 mg of the complex perfused the skin. Skin samples, taken before and after, were fixed in buffered formalin and embedded in paraffin. Five micron sections were taken, stained with rubeanic acid and counterstained with 1% Eosin Y solution. Histological examination showed that copper was present in all layers of the perfused skin. This work is relevant to the solubility of metallic copper in human sweat and to the possible therapeutic value of the "copper bracelet".


Essential fatty acid metabolism in cultured astroblasts. The fetal calf serum on which two astroblast cell lines were grown was shown to be deficient in essential fatty acids. The fatty acid profiles of lipids of these two cell lines showed very low amounts of polyunsaturated fatty acids. In order to know if this low level of unsaturation was due to the lack of essential fatty acids in serum, or to a lack of desaturase activities in the cells, we have investigated the modifications of the cell lipid fatty acid patterns when serum was enriched in essential fatty acids. Linoleic acid was incorporated in rather high amounts in the cell lipids, while linolenic acid was very poorly incorporated. These two essential fatty acids were converted into polyunsaturated fatty acids only when they were added alone to the serum. Both cell clones exhibited a lack in the delta 4 desaturase activity. No morphological changes of the cells occured after nine days of culture with modified serum.


Lipolytic activities against trioctanoin and monoolein in rat intestinal mucosa. The hydrolysis of trioctanoin and of monoolein by rat intestinal homogenates, catalyzed by the previously described "trioctanoin hydrolase" and "monoglyceride lipase" [Ref. 6 and 11], was shown to be entirely due to the action of the intestinal glycerol-ester hydrolase [Ref. 1 and 2] and to contaminating pancreatic lipase firmly bound to the intestinal cells. These conclusions are supported by experiments based on the chromatographic behaviour of the different lipolytic activities, the use of an immunoserum directed against rat pancreatic juice and the relief of bile salt inhibitions by pancreatic colipase. The results are in favour of the existence of a single lipolytic enzyme in the rat intestinal mucosa, i.e. glycerol-ester hydrolase, active against short- and medium chain mono-, di- and triglycerides and against long chain monoglycerides.


Twenty-four-hour patterns in human performance, subjective and physiological variables and differences between morning and evening active subjects. A study was undertaken to investigate differences between habitually morning active and evening active subjects as regards 24-hr patterns in physiological and subjective arousal and performance efficiency under conditions of 72 hr of sleep deprivation. The results were that very consistent 24-hr rhythmic were present in some of the performance variables, subjective alertness, oral temperature and urinary adrenaline excretion. No significant differences in 24-hr patterns could be detected between a group of habitually morning active and a group of evening active subjects. Significant correlations were obtained between oral temperature and some of the performance measures, while adrenaline excretion was not significantly correlated with performance. There were rather high correlations between subjective alertness ratings and some of the performances.


Event-related brain potentials in selective response. Event-related potentials were recorded during performance of a simple reaction task and two selective response tasks differing in the intensity of the non-key stimulus. Known differences in reaction between these tasks were confirmed, but there were no corresponding differences in amplitude of the slow potential shift which develops in the EEG in anticipation of the imperative signal. Since this anticipatory potential is not affected by event uncertainty, it seems to be related to motor preparation rather than to expectancy. Performance differences between simple and selective response tasks appear to depend not upon differential preparation, but upon selective processing of the imperative signal, which is reflected in the N120 component of the visual evoked response to that signal.


Attention and cardiac activity: heart rate responses during a variable foreperiod, disjunctive reaction time task. A variable foreperiod disjunctive reaction time task was used to evaluate the effects of stimulus discriminability on cardiac activity. In two sessions, 20 male undergraduates received easy or difficult auditory discrimination trials in blocked and random series. In the random session only, when the warning stimulus provided information about the difficulty of the subsequent discrimination, difficult trials were associated with greater foreperiod deceleration. Cardiac accelerations also tended to be greater in the foreperiods of difficult trials. There was an effect of both warning and imperative stimuli on the heart rate of the cycle in which the stimuli occurred which depended on the difficulty of the trial. The results are discussed in terms of the psychological significance of the different heart rate responses.


Feedback and instructional effects in the voluntary control of digital pulse amplitude. The effects of verbal instructions and these instructions plus visual analogue feedback on the voluntary control of digital amplitude were examined in 2 groups of 8 subjects over 8 training sessions. Control of pulse amplitude was demonstrated but feedback did not add consistently or reliably to the effect of verbal instructions. Control did not improve over the period of training nor did it become more specific, i.e. the effect did not become localised at the feedback site and pulse interval, respiration rate and forearm muscle activity were implicated in the response throughout. The results are interpreted as providing little support for the theory that learning to control a visceral response is analogous to learning a motor skill.


Circular dichroism of insulin A chain in water: trifluoroethanol mixtures. Use of linear and nonlinear least squares analysis. Circular dichroic spectra obtained for S-carboxymethylated insulin A chain in water: trifluoroethanol mixtures show that a marked conformational transition occurs as the concentration of trifluoroethanol is increased to (v/v) 83% to give a polypeptide containing about 43% of the residues in the alph-helical conformation. Several proposed methods of analysis, including both linear and two nonlinear lest squares methods, were unable to quantitate the amount of beta-structure present in the polypeptide in 83% trifluoroethanol. Examination of the methods of analysis lead to the conclusion that the current models for far UV CD analysis are not adequare for the data obtained in this study and shows that nonlinear least squares procedures may lead to erroneous conclusions.


On conformations of the superhelix structure. The slight deformation of a helical macromolecule leading to the superhelical structure is considered. General equations which connect "internal" stereochemical parameters of the backbone of a helical macromolecule with "external" parameters of the superhelix are obtained; they are analogous to those of Shimanouchi and Mizushima. The case when the radius of the major helix is much greater than the radius of the minor helix is treated. Assuming that all conformational changes are due to small distortions of the rotation angles (bond angles and bond lengths are kept constant) the general equations  reduce to a set of nonhomogeneous linear algebraic equations. Its solution (in the case of the DNA double-helix in B-form) shows that the DNA backbone can form a coiled-coil with parameters close to those estimated from experimental data on DNP in chromatine from nuclei of cells.


Resolvability of Adair constants from oxygenation curves measured at low hemoglobin concentration. Analyses of low concentration oxygenation curves for apparent Adair constants in which the effects of dimers is ignored have been explored using recently determined values of the overall energy coupling parameters. For high affinity systems and favorable energy distributions, it is found that the errors in estimated binding free energies may be less than one kcal provided the measurement errors are strictly random and of small magnitude. These errors are nevertheless quite substantial as compared with the differences between values for the successive binding steps.


Studies on trimethylsilyl derivatives of 1-alk-1enyl-2-acyl glycerols by gas-liquid chromatography mass spectrometry. Trimethylsilyl derivatives of 1-hexadec-1-enyl-2-decanoyl and -2-octadecenoylglycerols and 1-octadec-1-enyl-2-decanoyl and -2-octadecenoylglycerols were subjected to analysis by a gas chromatograph mass spectrometer system. The characteristic ions are as follows: (1) ions at [M-15]+ and [M-90]+- are observed, 2 or 16 mass units less than those of the corresponding 1-alkyl-2-acyl or 1,2-diacyl derivatives; (2) the base peak is m/e 129; (3) the ion at [M-R1CH=CHO]+ or [R2COO+130]+ is present in high abundance; (4) ions at m/e 311 and 339, characteristic of trimethylsilyl derivatives of 1-hexadec-1-enyl and 1-octadec-1-enylglycerols, are due to [R1CH-CHO-1)+73]+, also confirmed from studies on the perdeuteriotrimethylsilyl derivatives; (5) several ions based on one acyl group, [R2CO]+, [R2CO+74]+ and [R2COO+74]+ are also observed.


Determination of mass spectrometric sensitivity of different metalloporphyrin esters relative to porphyrin ester. Quantitative determination of metalloporphyrin contamination in preparations of biologically important porphyrins was achieved mass spectrometrically by application of the integrated ion current technique. For this purpose, the relative molecular ion sensitivities of the contaminating metal complexes were determined from the ratios of the integrated molecular ion currents of a series of calibration samples containing a porphyrin ester and one of its metal complexes in known molar ratio. Complexes formed with divalent ions of Cu, Zn, Fe, Co and Ni of copro- as well as uro-porphyrin permethylester were all found to have the same molecular ion sensitivities as their metal-free porphyrin ester. The relative metalloporphyrin ester content in a sample of porphyrin ester was thus obtained directly as the integrated ion current ratios of the normalized molecular ions. The preparation of calibration samples, including complex formation and the mass spectrometric methodology, is described and discussed.


Variance analysis of error in selected ion monitoring assays using various internal standards. A practical study case. This study contains a practical application of variance analysis for the evaluation of precision in selected ion monitoring using stable isotope variants or chemically related compounds as internal standards. Variance analysis is applied to determine the relative contributions of the errors due to long-term and short-term effects of instrument stability. The technique allows the resolution of variance due to sample manipulation from that introduced by the gas chromatography mass spectrometry instrumentation. This provides an objective measure of potential internal standards. Evaluation of the dependency of these errors upon concentration indicates appropriate weighing factors to be applied for regression analysis.


Identification of dihydrothiazoles in urine of male mice. The compounds 2-isopropyl-4,5-dihydrothiazole and 2-sec-butyl-4,5-dihydrothiazole were identified in urine of male mice. Their excretion in mouse urine is sex-dependent. In the urine of female mice the two dihydrothiazoles are either absent or present only in trace amounts. The analytical procedure includes the adsorption of the volatile mouse urine constituents on Tenax GC, their gas chromatographic separation and the detection of the sulfur compounds by the sulfur-specific detector. The identified compounds were synthesized. Natural and synthetic compounds had the same mass spectrometric properties and retention data.


Structural analysis by mass spectrometry of the major mammalian retinal ganglioside, a sialyl-sialyl-dihexosyl-ceramide. The major mammalian retinal ganglioside was analysed by mass spectrometry. Three types of derivatives were used, the fully methylated, the methylated and reduced and the methylated, reduced and trimethylsilylated compound. The mass spectra unambiguously showed the ganglioside to be a sialyl-sialyl-dihexosyl-ceramide. This is the first conclusive proof that the two N-acetylneuraminic acids are bound to each other constituting the terminal disaccharide. The major species contained sphingosine and stearic acid.


Precise measurement of total body water using trace quantities of deuterium oxide. This study was undertaken to investigate the possibility of measuring total body water in human subjects to better than +/-0.5%. Accurate serial estimates of total body water were required to complement densitometric and anthropometric measurements used to monitor body compositional changes in obese patients undergoing dietary or surgical weight reduction therapy. The method required the oral administration of 1-2 g of deuterium oxide and the analysis of pre-dose and respective equilibrated samples of urine, plasma or saliva. The sample size required for analysis was 5 microliter and the conversion of gaseous phase was accomplished using a uranium reduction furnace. Isotopic enrichment of samples was measured using a mass spectrometer incorporating several features designed to cope with problems inherent in H2/H2H isotopic analysis. Reproducibility of sample preparation and accuracy of the mass spectrometer were tested using international standards and shown to give an overall sensitivity of 2 parts in 10(7) for the determination of deuterium in H2O/H2HO mixtures. This precision has enabled us to demonstrate that isotopic fractionation of deuterium with respect to hydrogen occurs within the body and expands the potential use of this isotope for quantitative biochemical studies in the human subject.


Cyclic alkylboronates as derivatives for the characterization of cannabinolic acids by combined gas chromatography and mass spectrometry. Stable cyclic methyl- and n-butylboronates were prepared from the cannabinolic acids and used as derivatives for the characterization of these compounds by combined gas chromatography mass spectrometry. Retention times of the methylboronates were comparable with those of the trimethylsilyl derivatives. The mass spectra of these derivatives were more characteristic of the cannabinoid skeleton than were the spectra of the trimethylsilyl derivatives where fragmentation was associated with the trimethylsilyl groups. Molecular weights were lower and abundant molecular ions were obtained. These derivatives also offered the possibility of distinguishing between isomeric cannabinolic acids where only isomers possessing adjacent phenolic and carboxylic functions form cyclic boronates.


The measurements of propantheline ion in biological fluids after administering propantheline bromide to man. The measurement of propantheline ion has been accomplished in urine and plasma following administration of propantheline bromide to man. Trideuteropropantheline bromide is added to the biological fluid to act as a carrier and internal standard to quantify the propantheline ion using multiple ion monitoring. Methane was used as reactant gas but following the discovery of exchange of the trideuteromethyl group when using methane, ammonia was used for later analyses. The determination of propantheline ion in urine and plasma after administration of propantheline bromide to man is described.


Ionic channels of nerve: questions for theoretical chemists. Ionic channels are the voltage-dependent, ion-selective pores of nerve membranes. They are probably macromolecules which change their conformation as the electric field is changed. One conformation makes the membrane permeable to ions. Physiologists need the help of physical chemists and theoretical chemists to understand channels better.


Kinetic dielectric decrements and conducting liquids. We have calculated influence of ion migration on the measured dielectric constant of an electrolyte solution. The solvent is assumed to be a viscous, incompressible, polarizable liquid continuum having a finite dielectric relaxation time, while the ion is a rigid, impenetrable body. The decrement in the static permitivity of the solution with respect to the pure solvent is shown to be proportional to the product of solvent dielectric relaxation time and conductivity of the solution, with an additional contribution arising from viscous relaxation in the solvent.


Variational principles in nonequilibrium thermodynamics. The variational principles introduced by Onsager into nonequilibrium thermodynamics are discussed. Several new variational formulations of the linear laws are introduced including both a Hamiltonian and Lagrangian form. Also an Onsager-Machlup type variational description of fluctuations is shown to apply  at steady states even far from equilibrium.


Oscillatory chemical reactions: the Tomita-Kitahara model. Biological clocks share with excitable membrane (nerve, muscle) the requirement that the chemical systems that underlie them have unstable steady states, and hence are capable of limit-cycle oscillations. A particularly simple model of a chemical oscillator is discussed, especially with reference to the reverse reaction, which tends to damp the oscillations. For thermodynamic reasons (entropy production) reversibility can not, however, be neglected. The steady state, even when stable, does not minimize the dissipation function. Computer studies show the response to sinusoidal variation of the rate constant governing the autocatalytic step.


A model for the packing of cells.

A model for the packing of cells. A model is presented for the statistical and geometrical aspects of the packing of cells. The model is closely related to lattice polymer models and the Ising model of ferromagnetism. Fundamental questions of existence and analyticity in the infinite volume limit are discussed, and the results of some initial numerical studies are given.


Corynecin (chloramphenicol analogs) fermentation studies: selective production of Corynecin I by Corynebacterium hydrocarboclastus grown on acetate. Corynebacterium hydrocarboclastus KY 8835 grew in the acetate medium and accumulated 28mM Corynecins which was the highest production yield among the processes using various carbon sources. Selective production of Corynecin I (over 90% of all Corynecins), which had been desired for increase of the product yield, was achieved in this acetate medium. To keep the concentration of acetate, ammonium, and phosphate ions in the optimum range throughout the fermentation, a solution containing CH2COOH (50%), CH3COONH4 (9%), and KH2PO4 (0.2%) was fed continuously to the culture medium as the pH controlling agent. The addition of KCl (1%) and NaCl (1%) to the medium at 12 hr after inoculation stimulated the production of Corynecins.


Characteristics of unbuffered gel-immobilized urease particles. I. Internal pH. The overall rate of reaction of a gel-immobilized urease particle necessarily depends upon the hydrogen ion concentrations within the particle. When the particle is unbuffered, the internal hydrogen ion concentrations are a consequence of the local rates of reaction and the rate of egress of the products of hydrolysis. A simple apparatus has been devised which allows a fairly rapid determination of the hydrogen ion concentration in the center of a particle for any given size, enzyme concentration, substrate concentration, and external pH. The products of urea hydrolysis are self-buffering in the region of pH 8.83 and for an external pH less than the self-buffering pH, the pH within the particle is increased because of the reaction. When the external pH is greater than the self-buffering pH, the converse occurs. The pH at the center of the particle approaches the self-buffering pH with an increase in particle size and enzyme concentration. The external increase in the external substrate concentration has a limited effect, simply rendering the local rates of reaction to be of zero order. The center-line pH and therefore all internal hydrogen ion concentrations depend upon the parameter L square root pe and the external pH. Differences between the external and center-line pH values of the order of units are unexceptional. The implications of the internal pH profiles on the local and overall rates of reaction are explored.


Characteristics of unbuffered gel-immobilized urease particles. II. Overall rate of reaction. The overall rates of reaction of unbuffered gel-immobilized urease particles have been investigated with the aid of a packed-bed differential recycle reactor. Both substrate and enzyme concentrations have received attention. Cylindrical gel particles contained within impermeable tubelets were used to provide the physical strength necessary for the packed-bed arrangement and a one dimensional diffusion path to aid understanding of the complex interactions between substrate and product diffusion, and their effect on the reactions taking place. The experimental data have been interpreted with the aid of an enzyme rate equation (ERE) which relates the free solution characteristics of the enzyme to the conditions within a diffusion limited particle. The internal hydrogen ion profiles have been accommodated by a lumped parameter, the apparent pH (pHapp). Two methods have been suggested for the calculation of pHapp and the loss of activity on particle preparation, these methods are based on the use of the ERE in conjunction with experimental data.


Competition for mixed substrates by microbial populations. A model for the growth of an organism on multiple substrates was developed, assuming that each substrate has a competitive inhibition effect on the uptake of other substrates. The model was extended to examine mixed substrates, showing that the coexistence of several species at steady state in continuous cultures is possible, even when all the organisms all strongly prefer the one substrate. The diversity of nutrient sources in a real system may be a key factor in supporting a heterogeneous microbial population.


Comparison of existing classifications for coding congenital malformation and genetic syndromes. One thousand consecutive new registrations at the B.C. Health Surveillance Registry were coded by means of ICD (8th Edition), Cardiff Classification, and SNOMED systems. The Cardiff system uses the basic ICD number with important fifth and sixth digit modifiers, which improve discrimination. In certain conditions, however, the basic three-digit number differs from that in ICD and hence comparability is not always possible. The SNOMED system has six subcategories, followed by a five-digit code. These subcategories deal with Function, Disease, Topography, Etiology, Morphology, and Procedure. For our particular needs, the SNOMED system was not entirely satisfactory as it has not expanded sufficiently for many of the malformation syndromes. For hospital use, though, the SNOMED system may have numerous advantages over other existing systems.


[Generators of pathologically enhanced excitation as determinant structures in the spinal cord]. Using tetanus toxin as a tool to disturb the inhibition, enhanced excitation generators were created in the anterior horns of lumbar segments of the left and right sides of albino rat spinal cord. The generators worked at different regimens: the "left-side" generator originating under conditions of a more prolonged toxin action, on being activated by trigger stimulation, produced first tonic and then intermittent activity or periodic spontaneous discharges; the "right-side" generator produced only tonic activity. After the inhibition of one of the generators by glycine the other continued working in its regimen. Activation of one of the generators was followed by inhibition of the effect of the other. In case of separate activation of one of the generators all pools of the spinal and supraspinal motor neurons reproduced the activity pattern of the generator working at that moment. Thus, the latter played the role of the hyperactive structure determining the system behaviour, i.e. the determinant. The evidence obtained is analyzed from the point of view of the general concept on the role of the determinant structures in the nervous system activity and the theory of generator mechanisms of the neuropathological syndromes characterized by the system hyperactivity.


[Changes in the sensitivity of aortic baroreceptors under the influence of the sympathetic nervous system]. A dependence of the activity of the aortic baroceptors on the aortic pressure in stimulation of a branch of the stellate ganglion innervating the aortic baroceptor zone was studied in experiments on cats. The sympathetic stimulation failed to change the threshold of baroceptors (80 mm Hg) and their activity within the pressure range of 80-140 mm Hg. With higher aortic pressure the sympathetic stimulation decreased the baroceptor activity. As a result, an increase of the baroceptor activity with pressure was less and maximal activity was attained at higher aortic pressure (greater by an average of 32 mm Hg). The action mechanism of the influence of the sympathetic innervation on the baroceptor sensitivity, and physiological significance of this phenomenon are discussed.


[Effect of increased calcium ion concentration on the electrical and contractile activity of guinea pig ureter smooth muscle]. Experiments were performed on the smooth muscle cells of the guinea pig ureter by means of the double sucrose-gap method. An increase of Ca2+ concentration (22 mM/1) in Ringer-Locke solution causes hyperpolarization and a slight increase of the membrane resistance. The amplitude of the first spike potential and the plateau decreases, the amplitude of oscillations increases. In sodium-free Ringer-Locke solution when the action potential in the smooth muscle cells of the ureter turns into a simple spike potential, an increase of Ca2+ concentration causes a marked increase of the action potential amplitude.


[Influence of adaptation to altitude hypoxia and subsequent de-adaptation on the retention of a temporary connection]. Adaptation to simulated altitude hypoxia was followed by a significant increase of the degree of retention of active avoidance of the conditioned reflex. Twenty four hours after the reflex elaboration the adapted rats required only half of conditioned and unconditioned stimulus combinations in the reflex retention test than control ones. This effect was reproduced 1 and 5 days after the cessation of hypoxic training. The obtained increase in the degree of the conditioned reflex retention disappeared during deadaptation 10 and 27 days after the cessation of hypoxic training.


[Effect of conditioning to altitude hypoxia on mesenteric microcirculation in rats]. A long-term training of rats to high-altitude hypoxia (in an altitude chamber at the height of 6500 m, 6 hours daily, for 30 to 40 days) results in a significant hyperemia of their intestinal wall and mesentery. The number of functioning capillaries of trained rats exceeded that of control animals several times, but the blood flow was slowed down in the dilated vessels and their structure was altered. Besides an obvious increase in blood rachemia in the arteries and veins, its rarefaction is observed in the capillaries. Definite hemorheological changes, probably associated with polycythemia, and an increased hematocrite index were noted.


[Influence of latent tetanus poisoning on nervous system excitability in aseptic inflammation]. Experiments on rabbits demonstrated phasic changes in the nervous system excitability in aseptic inflammation against the background of latent tetanus intoxication (according to the results of investigation of the neuro-muscular chronaxia and the entire electroexcitability curve ("power-duration"). Apparently this is of definite significance in the changes of interrelations between the reactivity and the resistance of the organism and in the action on the phlogogenic agent.


[Growth and heat resistance of rats repeatedly exposed to heat during early ontogenesis]. The influence of a 10-fold heat training from the 1st to the 10th to the 20th, and from the 20th to the 30th days after birth on further growth and resistance of the animals to the high temperature was investigated. The growth and survival of rats was found to depend on the regimen of heat training. The animals aged 10 to 20 days proved to be most sensitive to the action of high temperature. Heat resistance of these rats turned out to be maximally increased at the period of primary sexual maturation, and the growth rate was retarded in comparison with the animals subjected to heat training at other age periods.


[Multi-component system of proteins binding estradiol in rat liver cytosol. Relationship to sex steroids]. Marked sex differences in estradiol-binding capacity ratio of components with Stokes radii (a) 7.0 and 2.5 nm in the liver cytosol of mature intact rats were found to be diminished, but not fully eliminated by gonadectomy. Prolonged administration of estradiol (50 microgram/animal for 8 days) to gonadectomized rats led to inhibition of the estradiol-binding activity of all the components in the liver cytosol of male and female animals. Administration of testosterone-propionate (2 mg/animal for 8 days) to gonadectomized animals stimulated selectivity the peculair estrogen-binding protein with a 2.5 nm not only in male, in which this protein was normally found, but also in female rats. It is supposed that sex differences in the system of estradiol-binding proteins in rat liver cytosol are determined by sex differentiation of the system at the early stages of ontogenesis, on the one hand, and by the active regulating influence of estrogens and androgens at the late stages of ontogenesis, on the other hand.


[Release of DNA from the nuclei of hepatocytes of rats following their treatment with heparin]. The influence of heparin on the protein fractions and DNA of the isolated nuclei of hepatocytes was studied by biochemical and cytospectrophotometrical methods on Wistar rats. It was found that 0.05% sodium heparinate solution blocked 95% of the histones, not influencing the non-histone proteins of the cell nucleus. Histone blocking was connected with the DNA release into the incubation medium in the amounts proportional to the incubation period.


[Resistance of different forms of rat liver cytochrome P-450 to influence destabilizing microsomal membranes]. The authors studied the effect of actions destabilizing the membrane of hepatic microsomes on the spectral properties of cytochrome P-450 (P-448) in rats--intact and those given phenobarbital (PB) or 3-methylcholantrene (MC). There was revealed a marked resistance of microsomes induced with PB and MC to the enzymatic and nonenzymatic peroxidation of polyunsaturated fatty acids of membrane phospholipids. There was a distinct difference in the sensitivity of cytochrome P-448 and cytochromes P-450 in rats--intact and those given PB--to desoxycholate-Na treatment in vitro. The results obtained pointed to structural changes of the microsomal membranes in the case of induction with PB and MC representing two different types of hepatic monooxygenases inductors.


[Space-time organization of systems of membrane hydrolysis and transport in rat small intestine]. Glucose transport by the concentration gradient with the incubation for 90 min in 0.2% glucose and soluble starch solutions was studied in Wistar rats in 5 segments of the small intestine by the "sac turned inside out" method. Serous fluid was completely replaced by a new portion of Ringer's solution every 15 or 30 min. Substrate load synchronized the enterocyte population and stabilized the transport systems. The changes of glucose absorption during the period of about an hour proved to differ in the 5 segments against the background of continuous and interrupted substrate load. These differences were due to the properties of the transported systems autocontrol and the reactivity level of the given enterocyte population. Areas with different reactivity were found to alternate along the intestine. Between the 8th and 16th hour (rats were sacrificed every 2 hours) starch glucose transport fell sharply in the proximal, and, to a lesser extent, in the middle segments. On the contrary, absorption between the 8th and the 12th hour was considerably intensified in the distal segments. The changes of the strach glucose transport during the period of about an hour along the intestine differed. The data obtained are discussed with consideration to the possible role of the undulating processes in the individual enterocyte population and in the small intestine as an integral system.


[Influence of thyroxine on lipid peroxidation in mitochondrial membranes]. In concentrations above 5.10(-7) M thyroxin brought about a delay in chemifluorescence development and decreased the slope of semilogarythmic plot of chemifluorescence curve at the initial step of the fluorescence slow flash. The antioxidant activity of thyroxin was found to be insignificantly lower than that of the well-known antioxidant alpha-tocopherol. In lower concentrations (about 1.10(-8) M) the hormone can act as prooxidant, the effect being the highest in the case of intact mitochondria incubated in the phosphate-containing medium in the presence of Mg2+ ions, and much less in the mitochondria treated in hypotonic medium containing Ca2+ ions.


[Uptake of gamma-aminobutyric acid by 3H-glial cells and synaptosomes of rat cerebral cortex under the influence of psychotropic substances]. Chlorpromazine was shown to inhibit non-competitively the 3H-GABA uptake both by the glial cells and synaptosomes; synaptosomal uptake was more sensitive to the inhibitor. Only the low-affinity GABA uptake in the glial cells was competitively inhibited by beta-alanine. On the whole, there was a correlation between the inhibition of GABA uptake by psychotropic drugs in the glial cells and in the synaptosomes. It is assumed that there existed two different systems of GABA uptake (of high and low affinity) in the nerve endings and glial cells.


[Effect of hydrocortisone on DNA synthesis by urinary bladder epithelial cells and renal cortical substance and on the concentration of glucocorticoids in the blood]. Single administration of hydrocortisone (10 mg per 100 g of body weight) led at first to increase, and then to statistically significant reduction of the peripheral blood plasma corticosteroid level in rats. An increase of glucocorticosteroids was accompanied by a reduction in number of DNA-synthesizing cells of the urinary bladder epithelium leading later to the fall in the number of mitoses, whereas reduction of the hormone--to increase of the cell change from G1 to S period. At the same time inhibition of the proliferative processes in the kidney epithelium lasted during the whole experiment (48 hours). Possible causes of different sensitivity of the epithelial tissues to hydrocortisone are discussed.


[Metabolism of immunoglobulin receptors by lymphocytes maintained in mouse spleen at the peak of the primary response]. Splenocytes obtained from mice on the 5th day after the injection of sheep red blood cells (SRBC) lost irreversibly approximately 50% of their surface immunoglobulins in the course of four-hour in vitro incubation. On the contrary, there were no changes in the total amount of surface immunoglobulins following in vitro incubation of splenocytes obtained on the 9th day after the antigen injection. Metabolism of antigen-binding receptors of immune splenocytes was studied by the method of rosette-formation. Incubation for 20 hours of splenocytes obtained on the 5th day after the SRBC injection demonstrated a loss of 70% of rosette-forming (RFC). The remaining RFC belonged to the theta-positive lymphocytes with the half-life of their antigen-binding receptors of approximately 4 hrs. Similar half-life was found for the RFC receptors of mouse spleen on the 9th day after the antigenic stimulation. No reduction of the RFC count occurred during their in vitro incubation for 20 hrs. It is supposed that a reduction in the number of the RFC obtained at the peak of the primary immune response could result from the inability of immune lymphocytes to synthesize new receptors or from blocking the newly-formed receptors by a soluble factor produced by immune lymphocytes following their in vitro incubation.


[Development of tumors in Unio pictorum bivalve mollusks under the influence of N-nitroso compounds]. Diethyl- and dimethylnitrosamines dissolved in tank water in doses of 200-400 ppm induced neoplasms of the digestive gland (basophilic cell neoplasms) in 16 of 95 and in 6 of 17 molusco which survived to the time of appearance of the first tumour (38-39 days), and also neoplasms of the hemopoietic system (lymphatic leukemia) in 7 and 1 molluscs, chiefly in combination with tumours of the digestive gland. Methylnitrosoguanidine induced only inflammatory changes at the site of injections. It is of expedience to use the molluscs as a biological indicator of the hydrosphere pollution with chemical carcinogens.


[Effect of estrogens on the concentration of carcinoembryonal antigen in the serum of rats with intestinal neoplasms and in the presence of non-specific injuries to its mucous membrane]. Carcinoembryonal antigen (CEA) was shown to be present in 70% of rats with tumours of the large intestine induced with 1,2-dimethyl-hydrazine and in rats with posttraumatic mucosa regeneration of the same organ. After treatment with estrogen--diethyl stilbestrol propionate (0.57 microgram/day for 6 days) the incidence of CEA finding in the serum of these rats was increased. In rats with an injured mucosa of the large intestine estrogen treatment prevented the natural decrease of CEA during the decline of regeneration.


[Effect of x-irradiation on DNA-synthesis in uterine gland epithelium and the relationship of that effect to the phase of the mitotic cycle]. The autoradiographic method with the use of thymidone-3H was applied to the study of X-irradiation influence on the DNA synthesis in the epithelium of the endometrial glands of ovariectomized mice stimulated with synestrol, and of the dependence of this action on the mitotic cycle phase. It appeared that in local irradiation of the uterus with a dose of 400 r reduction of the labeled nuclei index (LNI) in the cells of the endometrial glands differed and depended on the mitotic cycle phase of the majority of the cells.


[Protein synthesis during the period of mitotic recovery following exposure to colchicine and colcemide]. Autoradiographic study with leucine-3H showed the restoration of normal mitosis course after the action of colchicine on the culture of Chinese hamster cells to be accompanied by a marked activation of the protein synthesis exceeding the initial level. This intensification of protein synthesis precedes complete normalization of the mitotic activity in the cell culture. The course of normalization under the influence of colcemide is characterized only by the stabilization of the protein synthesis at the initial control level.


[Repression and derepression of lactate dehydrogenase loci during mouse development]. The ultramicroelectrophoretic method was applied to the study of the lactic dehydrogenase (LDH) spectrum alterations during the developmental phases of mice: ovulated ova--zygote--blastocyte--embryo--oocyte--ovulated ova. Only H-subunits were found in the embryo cells before the 5th day of development. After this M-subunits appeared indicating derepression of LDH-A locus. On the 8th day of the embryonal development deres pression of the LDH-B locus was observed to disappear during the oogenesis, being the result of progressive repression of locus LDH-A. Dictiotena of meios prophase is characterised by active H-subunit synthesis and a gradual decrease of the M-subunit synthesis. During the whole dictiotena phase the LDH-spectrum of the follicular cells was of the M-type.


[Morphofunctional characteristics of pacemaker formations situated in the atrioventricular valves of the heart]. By the microelectrode technique and the method of electron microscopy multicellular formations responsible for generation of the automatic pacemaker activity were found in the cusps of the atrioventricular valves in the rabbit heart. Cells of different morphological structure were revealed: P-cells, cells of the Purkinje type and transitional cells. These formations have rich innervation, mainly of cholinergic nature. A certain correlation was established between the morphological and electrophysiological characteristics of these cells.


[Electron-autoradiographic analysis of projections of somatosensory cortex regions I and II to the posterior ventral nucleus of the thalamus]. To reveal the differences in the nucleus ventralis postero-lateralis (VPL) of the thalamus of the corticofugal fibers originating from the first (S1) and the second (S2) somatosensory areas a combined morphological study was undertaken using autoradiography and electron microscopy of the degenerating terminals. Within the range of VPL distribution of the cortico-fugal fibers of the corresponding zones of two cortical somatosensory regions differed from one another. The terminals of both types of the fibers established synaptic contacts, chiefly with the distal dendrites of the relay VPL cells, and much more rarely with the dendrites of the interneurons of the Golgi II type. No direct convergence of the fibers originating from two somatosensory regions on single VPL elements was observed.


[Ultrastructural mechanisms of serotonin demyelination]. The effect of serotonin on dog white matter ultrastructure in the CNS was investigated. The intracysternal injection of this amine (6 microgram in 0.1 ml of physiological saline) induced significant lesions in the myelin and glia of the spinal cord white matter areas adjacent to the cerebrospinal fluid tracts. A loss of the regular structure and stratification of the myelin lamellae, as well as rupture and lysis of the myelin membranes, and their demyelination occurred. There was vacuolar dystrophy of oligodendrocytes; the astrocytes remained practically unchanged. After local intracranial injection of this amine (2 microgram in 0.01 ml of physiological saline) similar lesions were observed in the white matter of the cerebral hemisphere; in addition signs of inflammatory reaction were revealed at the late period of investigation. In control animals injected with physiological saline the changes were observed later and in gliocytes only. A conclusion was drawn that serotonin was capable of injuring the myelin and glia.


[Reactions of cardiomyocytes of the right half of the heart to trauma to the left]. Myocardial infarction of the left ventricle was induced in adult rats weighing 120-160 g by ligation of the left coronary artery; the left atrium was injured in the second group of rats; the third group of the animals was subjected to the sham operation (only the pericardium was removed). On the 5th postoperative day the dividing myocytes were found in the right auricle (mitotic index varied from 0.7 to 8.8%) and in the subepicardial region of the right ventricle (mitotic index from 0.8 to 2.9%). On the third postoperative day the myocytes of the myocardium of senescent rats weighing 300-430 g proliferated in the left auricular appendage (mitotic index--from 1 to 5.1%) and in the right auricula appendage (in one of 8 cases, mitotic index 4.2%). Only rare mitoses were found in the subepicardial region of the left ventricle.


[Autoradiographic study of DNA synthesis in rats with experimental myocardial infarct]. Rats with experimental myocardial infarction were injected with 3H-thymidine; DNA-synthesizing nuclei were counted in different parts of the heart. Myocardial infarction enhanced insignificantly the DNA synthesis in the nuclei of myocytes located at the periphery of the injured zone. However the label localization in the myocyte nuclei was rarely undoubtful. There was no increase in the number of labeled myocyte nuclei when a higher dose of 3H-thymidine was used or its injection was repeated thrice. Proliferation of connective tissue cells was activated in all the parts of the heart. The number of the connective tissue DNA-synthesizing nuclei rose within 24 hours, reached its peak in 48 hours and remained at a higher level, as compared to control throughout the experimental period. On the first day the highest count of labeled stromal nuclei was found at the periphery of the infarction and on the third day -- within the injured zone. This indicated the growth of granulation tissue from the periphery to the center of the infarction focus.


[Histochemical and morphometric changes in the adrenal cortex under conditions of acute vascular insufficiency]. Morphofunctional changes in the adrenal glands of 50 male cats under experimental vascular insufficiency were studied by means of morphometric, histochemical, and biochemical methods, with a preliminary investigation of the histoenzymatological profile of the adrenal glands in intact animals. A decrease in the adrenal function was revealed five hours after induced hypotony resulting from intravenous injection of arfonade. Adrenal insufficiency, particularly that of the glucocorticosteroid function, under conditions of developing vascular failure can be considered as a peculiar adaptation of the organism to preserve its viability.


[Microspectrophotometric characteristics of renal enzymes under conditions of acute vascular insufficiency]. The activity of optic density of preparations of cat kidneys by studying lactic, succinic-, glycoso-6-phosphoric-beta-oxybutyrate, NAD.H2-, NADP.H2-tetrazolium-reductases, alkaline and acid phosphatases, was investigated in intact animals, those under hexenal anesthesia, under conditions of a 5-hour hypotension caused by administration of a ganglioblocker (arphonade), 24 hours after the termination of artificial hypotension and in profuse blood letting. Analysis of the optic density distribution curves showed that under conditions of acute vascular insufficiency all the nephron parts reacted by different changes of the enzymatic activity. These changes clearly demonstrated a sharp fall of the energy importance of the main oxidation way (Krebs cycle) and of the prevalence of glycolytic way and pentose shunt under conditions of circulatory hypoxia.


[Changes in phagocytizing and lymphoid elements in obstructive pneumonia]. A comparative cytochemical study of the state of phagocytizing and lymphoid elements was carried out during the action on them of a foreign body, under septic and aseptic conditions. Local and systemic changes of the phagocytozing and lymphoid elements ratio, permeability disturbances of the lysosomal membranes in them, an increase in destroyed macrophage, neutrophil, and lymphocyte count, serving as one of the main mechanisms of the developing inflammatory-destructive processes, were regard as indices of cell regression.


[Ultrastructural localization of thyrocyte acid phosphatase in the presence of thyrocyte hyperfunction in rats]. Acid phosphatase was found by electron microscopy in the lysosomes which appeared in great numbers in the follicular cells of rat hyperplastic thyroid gland. The other types of granules (mature secretory granules and lipids) whose amounts were also greatly increased in cases of functional thyrocyte strain were also nonreactive. The lysosomes were subdivided into three main groups according to distribution of the reaction product: the lysosomes with dense homogeneous deposit and with deposit in the form of densely or loosely packed dark round granules. The lysosome heterogeneity was apparently connected with their different functions also found within the colloid droplets in the form of inclusions of rarely located dark granules. The authors believe such granules to be the result of the merging of the colloid droplets and lysosomes. The acid phosphatase of the latter participated in the hydrolysis of the product of cell secretion with the formation of active substances.


[Relationship between the mosaicism of myocardial damage and the metabolic heterogeneity of myocardiocytes]. Uridine-3H (2 muCi/g of body weight) was injected intraperitoneally to albino mice; this was followed (in one hour) by isopropylnoradrenaline (0.1 mg/g of body weight). The animals were sacrified in 10 minutes. Autoradiographic analysis and polarisation-microscopic study of the myocardial section showed that myokardiocytes with a lower level of the RNA synthesis were to the contracture lesions.


[Macrophage-fibroblast interaction and its possible role in regulating collagen metabolism during wound healing]. The role of macrophages in the resorption of denatured collagen in the wound tissue was demonstrated by transmission and scanning electron microscopy investigation. Tight cell contacts between the macrophages and fibroblasts in the granulation tissue preceded active collagen synthesis and fibrillogenesis. A hypothesis was suggested according to which these contacts played an important role in the transfer of specific signals for the initiation of collagen biosynthesis; collagen decomposition products were considered as the actual signals, thus connecting the collagen catabolism and biosynthesis. Such a feed-back mechanism probably served as an integral part of the general hemostasis regulating the growth of connective tissue.


[Morphometry of the adrenals].

[Morphometry of the adrenals]. The authors report on the method of determination of the weight indices of the adrenyl gland glomerular, testicular-reticular and medullar zones with a spheroid shape; it is substantiated by mathematical analysis of a plasticine model of the adrenal gland, whose characteristics approached the actual ones. The method was particularly accurate in determination of the weight of the fascicular-reticular and glomerular zones, and less--in determination of the weight of the medullary layer, the method's error being 0.6-0.9% in the first case, 2.7-3.5% in the second and 5.3-6.4 in the last.


[Method of resistography].

[Method of resistography]. To eliminate the error of the resistograph connected with the animal's arterial pressure fluctuation an automatic stabilizer of the entrance pressure provided with an electromagnetic valve is proposed. To switch off the resistograph pump automatically when the blood flow into the apparatus is either hindered or stopped the following device is proposed: the pressure stabilizer and the servo system are controlled by electro-contact Hg manometers connected with the admission channel of the resistograph.


[Afferent projections of the aortic nerve onto different portions of the hypothalamic region]. Reactions of the neurons of the anterior, middle, and the posterior hypothalamus in response to electrical stimulation of the aortic nerve were studied. Two groups of neurons were distinguished in the pressor zones of the hypothalamic region. Changes of the neuron pulse activity preceded the reduction of the arterial pressure arising in response to the aortic nerve stimulation. Changes in the activity of the neurons of another group followed the arterial pressure reduction. A conclusion was drawn that excitations caused by stimulation of the aortic depressor nerves and the afferent influences, coursing during the depressor reaction from the baroreceptors of the vascular bed, spread to the neurons of the supraoptic, paraventricular, and the ventro-medial nuclei of the hypothalamus, the anterior and the lateral hypothalamic field, and the supramamillary region.


[Effect of micropolarization of focal and extrafocal brain structures on experimental epilepsy]. The influence of micropolarization (MCP, 0.05-0.75 muA) was studied in chronic experiments on rabbits with epileptogenic foci provoked by penicillin (doses of 500 and 1000 U) injection into the motor cortex. A single MCP of extrafocal structure (the corpus calossum, nucleus caudatus) and the site of the focus inhibited the development of seizure reactions. Numerous MCP of the focus facilitated the seizure reactions. These effects may be explained not only by the specific role of the structures studied in the processes of the formation and propagation of the excitation, their initial functional state, but also by the dependence of the activity of these structures on the MCP regimen.


[Influence of convulsive and pre-convulsive components of an audiogenic seizure on the process of consolidation of temporary connections]. In comparing the seizures of different severity in rats an amnestic effect of the convulsive phases of the audiogenic attacks on the short-term (in difference from the long-term) memory and the process of consolidation of temporary associations was revealed. Without behavioristic seizures the audiogenic attacks almost failed to derange the short-term memory. The results obtained confirmed the authors' results obtained earlier on different nature of the convulsive and preconvulsive phases of the motor activity during the audiogenic attack in rats. The preconvulsive phase is regarded only as a peculiar motor aura of the audiogenic attack without any convulsive components.


[Mechanism of disorders of inhibition of electrogenesis in spinal alpha-motor neurons in experimental local botulin poisoning]. Disorders of postsynaptic inhibition and of the spinal cord alpha-motoneurons were studied in cats with experimental local botulinum intoxication. A significant decrease of the reciprocal, and, to a lesser extent, of polysynaptic inhibitory postsynaptic potential (IPSP) was noted. With the appearance of total paralysis of the muscles in the poisoned extremity there proved to be an even greater depression of the reciprocal and polysynaptic IPSP; however, they never disappeared or turned into depolarization potentials. Synaptic permeability of motor neurons as a rule decreased during the IPSP development, this indirectly indicating a reduction of ion transport.


[State of neuromuscular transmission in rats with experimental hypoparathyroidism]. Experiments on albino rats with hypoparathyrosis demonstrated disturbances of the neuro-muscular transmission characterized by a decrease of the liminal strength of irritation in case of indirect stimulation, reduced amplitude of the muscle action potential, diminished latent period of response and absolute and relative refracterity phases and the character of muscular retention as well in indirect rhythmic stimulation. The investigated parameters displayed the same indices as in the control experiments following intravenous injection of proserine. Disturbances of the neuro-muscular transmission proved to be related to the presynaptic defect and could play a definite role in the development of the motor failure under conditions of hypoparathyrosis.


[Reserve production of renin by glomerular mesangial cells following experimental decrease in renal circulation]. The aorta was partially constricted between the origins of the two renal arteries in rats. The renin activity was studied separately in the capillary parts of the isolated glomeruli and in their arterioles. The renin activity was found in the capillary parts of the glomeruli 3 to 4 weeks after the constriction. The rough and smooth endoplasmic reticulum and the Golgi complex of the mesangial cells were activated at this period. The authors consider these results as a confirmation of the hypothesis on the reserve renin production by mesangial cells under prolonged decrease of renal circulation.


[Biosynthesis of renal prostaglandins in spontaneously hypertensive rats]. Biosynthesis of prostaglandins E2 and F2alpha from the exogenously added 14C-arachidonic acid in the renal medulla of spontaneously hypertensive (SHR) and normotensive Wistar rats (1 1/2- and 3 1/2-months-old) was studied. Investigations carried out showed that the formation of PgE2 from the arachidonic acid by the kidneys of SHR was significantly lower than in Wistar rats, whereas biosynthesis of PgF2alpha was practically normal. No difference in the prostaglandin-synthetase activity was noted in the kidneys of SHR of different age, whereas Wistar rats displayed a reduction of the PgE2 synthesis by the age of 3 1/2 months.


[Functional deficiency of erythrocyte thromboplastin factor]. A study of human erythrocytes in healthy and sick persons showed changes in the erythrocytes procoagulant activity, namely a reduction of the whole cell thromboplastin activity despite a sufficient procoagulant factor content. A term "functional insufficiency of thromboplastin factor of erythrocytes" or briefly "erythrocytic thrombopathy" is suggested for the above-mentioned changes of erythrocytes, similar with the changes of platelet procoagulant properties in one of the thrombopathy forms.


[Mechanisms of compensating the evacuatory function of the stomach after exclusion of the pyloric sphincter]. The rate of solid food evacuation from the stomach was investigated in chronic experiments on dogs with the stomach and the duodenum (its first part) fistulae. It was found that in exclusion of the pyloric sphincter (operations of pyloroplastics or gastroduodenostomy) the stomach evacuation was mainly compensated. However, judging by the free passage of rubber balls (3 mm in diameter) into the duodenum no structures replacing the pyloric sphincter formed in the stomach. The compensation of the stomach evacuatory function under these conditions was provided by a significant increase of the enterogastric reflex.


[Transformation of mitochondrial monoaminoxidases types A and B]. Chlorgyline, a selective monoamine oxidase (MAO) of the A type inhibitor, contrary to deprenil--MAO of the B type inhibitor, prevented the appearance in the rat liver mitochondrial membrane fragments incubated under aerobic conditions of qualitatively new properties to deaminate histamine or AMP. Apparently the MAO of A type but not of B type underwent a qualitative alteration (transformation) of catalytic properties under conditions favourable for partial oxidation of SH-groups.


[Latent activity of liposomal creatine phosphokinase]. Quantitative regimen for the entrapping of creatine phosphokinase (CPK) under different experimental conditions was investigated for the preparation of liposomes with a high internal protein content. As revealed, introduction of 20% of positively-charged lipid (stearyl amine) into the lipid mixture and subsequent one-minute sonication resulted in maximal levels of latent enzyme activity. Maximal entrapment (6.1%) was yielded when 300 mM glucose solution (pH 7.0) AND 0.5 MG OF PROTEIN PER ML WERE USED. Liposome preparations when washed twice at 205 000 g with physiological saline showed minimal external enzyme activity. During storage of liposomes for 7 days under argon protection the internal protein content has not diminished, nor the external activity has increased.


[Effect of sub-extreme and extreme factors on liver lysosomes]. Functional state of the lysosomal liver system in rats under subextreme and extreme influences (an intense muscle tension, fasting and combination of the both factors) was studied. An elevation of the acid hydrolases activity and labilization of the lysosomal membranes could be observed in the course of the body overstraining. Activation of lysosomal system correlated well with the extent of overstraining and reflected an adaptive reconstruction of the cellular structure and metabolism.


[2 possible mechanisms of action of thyroxine on mitochondrial swelling]. The effect of thyroxin on different types of mitochondrial swelling (induced by calcium, detergents, valinomycine, or lipid peroxidation) was investigated. Under conditions favourable for phospholipase-induced mitochondrial swelling on account of phospholipase activation (low Mg2+ concentration, and Ca2+ present in the incubation medium) the hormone proved to accelerate the swelling of the organellae. When phospholipase-induced mechanism of mitochondrial swelling was inhibited (high Mg2+ concentration) thyroxin depressed considerably the swelling of organellae induced by lipid peroxidation due to its antioxidant action.


[Effect of morphine and azidomorphine on the neuronal activity of the cerebral cortex]. Acute experiments were conducted on unanesthetized curare-treated cats and rats. Analgetic doses of morphine and azidomorphine proved to depress the spontaneous and bradykinin-induced neuronal activity in the sensory-motor zone of the cerebral cortex. The inhibiting effect of the drugs was eliminated by nalorphine. It is supposed that the depressive action of morphine and azidomorphine may be connected with their direct influence on the cerebral cortex.


[Changes in the large granular vesicles in sympathetic nerve endings under the influence of several pharmacologic agents]. Electron microscopic study of large dense-core vesicles of the adrenergic nerve endings in the middle cerebral arteries of some vertebrates showed substantial changes of the vesicles studied (in comparison with control) 60 to 70 minutes after rejections of ipraside (3 mg/kg) and dopamine (40 mg/kg). A statistically significant increase of their quantity was also revealed. Large dense-core vesicles are supposed to take part in the process of accumulation and transformation of noradrenaline and/or its precursors.


[Effect of aldosterone, vasopressin and natriuretic factor on the process of absoprtion in the rat small intestine]. Experiments were conducted on rat jejunum pouches turned inside-out. On addition of blood plasma containing the natriuretic factor there was a reverse relationship between the natriuretic factor concentration and the processes of absorption in the jejunum. No such effect was noted under the effect of aldosterone and vasopressin.


[Immune lymphocyte inhibition of activation of DNA synthesis in mixed cultures of normal lymphocytes in vitro]. Substitution of normal lymphocytes by immune ones as responding cells in the "one-way" mixed lymphocyte culture (MLC) results in inhibition of the DNA synthesis. This phenomenon is reproduced irrespective of the mouse strains used, immunization method (in vivo or in vitro) and various MLC techniques (macro- or microcultures, the presence of mercaptoethanol in the medium, control variants). Unlike the cytotoxic effect of immune lymphocytes showing a sharp peak on the 8th day after in vivo immunization, inhibition of the DNA synthesis activation in MLC occurred as the second day, and persisted for several weeks after immunization; this inhibition was reproduced on addition of a small fraction (5 to 10%) of immune lymphocytes to normal ones.


[Transplancental effect of ortho-aminoazotoluene on organ cultures of embryonal liver from mouse lines C57BL and CBA]. Transplacental influence of ortho-aminoazotoluol (OAAT) on the organ cultures of embryonal liver of highly-hepatomous mice CBA, and low-cancerous mice C57BL was the subject of this research. Transplacental influence of 12 mg OAAT caused an increase of embryonal liver organ cultures viability. Under the transplacental influence of 24 mg OAAT on CBA mice the toxic effect noted at the early cultivation period was replaced by the 20-25th cultivation days by the growth-stimulating effect. The results of in vivo experiments corresponded to the results of in vitro observations.


[Changes in the processes of bone marrow cell proliferation and differentiation in white mice under the influence of heparin]. In experiments carried out on albino mice daily heparin injections (250 units/kg) for two weeks resulted in 30% increase of leukocyte count in the venous blood. The femur bone marrow kariocyte count remained unchanged, while the leukoerythroblastic ratio rose from 4.1 to 7.6. Autoradiography demonstrated that the duration of the mitotic cycle and its phases in myeloid cells underwent no change, constituting: T--13 hrs, ts--8 hrs, tG2--1 hour, t(G1+M)--4 hrs. In heparinized animals the curve of labeled mitoses was nearly ideal. It is suggested that there occurred a change in the stem cells differentiation towards the myeloid type in the bone marrow under the effect of heparin.


[Division of stellate reticulocytes in rat liver following vagotomy]. The authors studied the 24-hour variations of mitoses in the stellate reticuloendothelial intralobular blood capillaries of rats under normal conditions and after bilateral subphrenic vagotomy. The greatest number of mitoses was noted in all the animals in the morning hours--3 to 7 a.m. A gradual decrease in the number of mitoses occurred in vagotomized and laparotomized animals by 10 p.m. After vagotomy the number of mitoses in the stellate reticuloendotheliocytes and in hepatocytes was double that in control; the character of the curve reflecting the 24-hour periodicity of mitoses in denervated liver was unchanged in comparison with control.


[Effect of sigetin on anovulation in rats]. The effect of sygethin on anovulatory cycle was studied in mature female rats. Anovulation was experimentally induced in rats on the one hand by stress effect (keeping rats in crowded conditions), and on the other hand by removing one ovary (hemicastration). Sygethin therapy (10 mg/kg for 4-5 days) proved to restore normal ovulation. When higher sygethin doses were used, and when it was administered for longer periods (for 2 to 3 weeks) no such effect proved to ensue. The results of these experiments suggest that small doses of sygethin stimulated luteinizing hormone secretion.


[Dynamics of changes in anomalous human cells during prolonged cultivation in the stationary phase. Trisomy 7 cells]. Ontogenetic changes of normal diploid cells and cells with chromosomal aberration (strain LHC-162; 47, XY, +7) were studied in prolonged cultivation in stationary phase. Cultivated cells were human found to possess their specific type of ontogenetic changes for each culture; the dynamics of these changes depended on the density of cellular population. Two morphologically different cellular subpopulations differing by the degree of cell nucleus heterochromatinization (normal strains) and three different cellular subpopulations in the aberrant strain (47, XY, +7) were discovered in studying the cell nucleus.


[Autoradiographic analysis of RNA synthesis by sympathetic neurocytes from a population depleted of nerve cells]. Autoradiographic analysis of the changes of the nuclear and cytoplasmic RNA in the sympathetic neurocytes of presenile mice subjected to partial desympathization during the first days after birth by guanetidine injections was carried out. As compared with the control mice of the same age, neurocytes of the populations with a sharply reduced number of cells were characterized by a slower migration rate of the newly-synthesizing RNA from the nucleus to the cytoplasm. The neurocytes of partially desympathized infantile, young and presenile mice were not larger than those of the control mice.


[Nucleic acid concentration in the brains of inbred rats of different strains]. The changes of nucleic acids content in the formation of conditioned skin-defence reflex was studied on inbred rats of various strains. In WGA rats the reflex was formed on the average aight stimulations. An increased RNA content was discovered by cytophotometric determination of the nucleic acids content in the neurons and perineural glia of the cerebral cortex in August rats only. It is assumed that an increased RNA synthesis in the animals of this strain can serve as the basis for their better training.


[Electron-microscopic autoradiography of RNA synthesis in the myocardium after damage to it]. Thermic burn of the wall of the left cardiac ventricle was inflicted to new born rats. Twenty-four hours after the injury the RNA synthesis of the myocardial cells remote from the site of burn were investigated by electron-microscopic autoradiography. Tissue samples were fixed 2 and 6 hours after the 3H-uridine injections. As compared with the control, experimental animals displayed a reduction of silver grains density over the nucleus and the cytoplasm of cardiomyocytes.