Electron microscopical proof of the presence of sugar beet yellows virus particles in the roots of sugar beet

The presence of beet yellows virus (BYV) particles was electron microscopically proved in the roots of sugar beet. Specimens for the electron microscopical examination of root sap were prepared by differential centrifugation. It was proved that, contrary to expectations, examinations in spring showed most virus particles in the basal part of the root. At the same time it was found by experiment that the diagnostical BYV antiserum, for which the antigen was prepared from sugar beet leaves, did not react with a purificate of BYV containing virus particles.     

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

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

Morphological Study of the Role of Calcium in the Assembly of the Rotavirus Outer Capsid Protein VP7

Maturation of rotavirus occurs in the endoplasmic reticulum (ER), a site of intracellular calcium storage. It was demonstrated previously that calcium plays an important role in the maturation of bovine rotavirus. We used protein A colloidal gold conjugated to an antibody to localize VP7, the outer capsid protein of the simian rotavius SA11, in permeabilized infected cells in the presence and absence of calcium in the culture medium. In medium containing calcium, VP7 was associated with nonenveloped double-shelled particles and membranous structures of the ER. In calcium-free medium, gold particles were not associated with the ER or with virus particles. Gold particles were distributed through the cytoplasm and were mainly associated with granular structures, but did not assemble onto virus particles. Our data suggest that in calcium-free medium, VP7 is synthesized, but does not remain incorporated, in the ER.     

Poly a blocks in the nuclear ribonucleoprotein complexes,containing pre-mRNA

Poly A was found in nuclear particles containing pre-mRNA. It was shown that during the isolation of 30S particles from rat liver or Ehrlich ascites carcinoma nuclei, all poly A is detached from the particles containing pre-mRNA and is found in the form of RNP with a sedimentation coefficient of about 14S. When RNP polyparticles are isolated in the presence of RNase inhibitor poly A is distributed among the particles of higher molecular weights.Since the sedimentation properties and buoyant density of the poly A-containing particles are different from the 30S particles it was suggested that the poly A fragments are bound not with informofers, but with another kind of protein.     

Analysis of the mechanism of intensification of fermentation process using yeast cells in a suspension of high-dispersed oxides

The differential microcalorimetry was used to explore an influence of particles of silicon dioxide, and also other high-dispersed oxides (0.05% of masses.) in water suspension of yeast cells on intensification of the process of their fermentation in endogenous metabolic conditions. It was shown that intensification of the processes of the vital activity of yeast microorganisms was observed in the specified interval of the concentration of silicon dioxide hydrosol particles. Mechanisms of interaction between SiO₂ particles and a surface of a cellular organism, as well as interaction between SiO₂ particles and one of metabolism products — carbon dioxide were studied. It was found out, that Al₂O₃, TiO₂ hydrosols also had a stimulating effect, but it is lower compared to that of SiO₂.     

Influence of the Chitosan Oligomer on the Phage Particles and Reproduction of Phage 1-97A in the Culture of Bacillus thuringiensis

The causes of bacteriophage 1-97A inactivation by the chitosan oligomer with a polymerization degree of 15 and the influence of the oligomer on the phage reproduction in the culture of Bacillus thuringiensissubsp. galleriae, strain 1-97, were studied. The study of the inactivation kinetics showed that, in 1 h, virtually all chitosan was bound to the phage particles, causing, as evidenced by electron microscopy, DNA release from the phage head, destruction of the phage particles, and agglutination of the phage particles or of their tails in the region of the basal plate. High-polymeric chitosan caused more pronounced destruction of the phage particles than the oligomer. It was established that chitosan prevented the production of complete phage particles. One of the mechanisms of such an influence may be the production in the presence of chitosan of phage particles devoid of DNA.     

Chondrocyte acting as phagocyte to internalize polyethylene wear particles and leads to the elevations of osteoarthritis associated NO and PGE2

It remains a mystery about the role of chondrocyte or cartilage on the co-existence of ultra-high molecular weight polyethylene (UHMWPE) wear particles from partial joint arthroplasty. An inverted co-culture system was performed to investigate the interactions between chondrocytes and UHMWPE wear particles. It was first time observed that chondrocytes can engulf UHMWPE particles and release osteoarthritis associated pro-inflammatory factors. TEM observation and flow cytometric analysis demonstrated the phagocytosis of particles by chondrocytes. It was found that polyethylene particles may reduce the viability of chondrocytes, and enhance the secretion of nitric oxide (NO) and PGE₂. In conclusion, all these phenomena may contribute to further cartilage degeneration after partial joint arthroplasty surgery. It is first identified in this study that the chondrocyte acts as phagocyte to internalize wear particles and leads to the elevations of precursor mediators of osteoarthritis.     

Changes in the size of reconstituted high density lipoproteins during incubation with cholesteryl ester transfer protein: the role of apolipoproteins.

It has been reported previously that the particle size distribution of discoidal, reconstituted HDL (r-HDL) changes dramatically during incubation in vitro with cholesteryl ester transfer protein (CETP). The present study was undertaken in order to determine whether these changes are influenced by the apolipoprotein composition of the r-HDL. Two preparations of r-HDL that contained egg phosphatidylcholine (egg PC) and unesterified cholesterol (UC) but differed in their apolipoprotein composition were used for the study. One preparation contained apolipoprotein (apo) A-I only (A-I w/o A-II r-HDL) while the other contained apoA-I and apoA-II (A-I w A-II r-HDL). The Stokes' radius of the major population of particles in the (A-I w/o A-II) and (A-I w A-II) r-HDL was, respectively, 4.8 and 4.9 nm. When the (A-I w/o A-II) r-HDL were incubated with CETP, most of the particles of radius 4.8 nm were converted to populations of smaller and larger particles. The smaller particles had Stokes' radii of 4.3 and 3.9 nm. The radii of the larger particles ranged from 8.2 to 13.7 nm. When the (A-I w A-II) r-HDL were incubated with CETP larger particles (Stokes' radii = 8.4-11.0 nm) appeared but there was minimal conversion to smaller particles. In addition, a significant proportion of the original (A-I w A-II) r-HDL of radius 4.9 nm was still present at the end of the incubation. These results are consistent with apoA-II inhibiting the conversion of r-HDL to small particles. It is concluded that the apolipoprotein content of r-HDL is an important determinant of the sizes of the particles that are formed during incubation with CETP.     

Determination of the amount of small messenger ribonucleic acid-containing particles free in liver cytoplasm.

To assess the contribution made by mRNA-containing particles to the heterogeneity previously observed among rat liver 40S ribonucleoprotein particles, the amount of poly(A)-containing RNA in subribosomal particles was determined. RNA was labelled with orotate in vivo for 24h and then for 50min. Poly(A)-containing RNA was trapped on filters impregnated with poly(U). Very little poly(A)-containing RNA was found in conventionally prepared ribonucleoprotein particles after fractionation in sucrose. However, after preparation of ribonucleoprotein particles by sedimentation through 1 M-sucrose in the presence of 0.15M-KCl or by precipitation with Mg2+ as described by Leitin & Lerman [(1969) Biokhimiya 34, 839-849], amounts of poly(A)-containing RNA were similar to amounts of mRNA found by other workers in total ribonucleoprotein particles. Even in such preparations, less than 5% of the total rapidly labelled RNA in native subribosomal-particle fractions was mRNA. It seems that mRNA-containing particles make up only a very small part of the population of subribosomal particles in liver.     

Characterization of lipoprotein particles isolated from the Golgi apparatus of rat liver

It has been proposed that particles within tubules and vesicles of the Golgi apparatus of liver cells are precursors of very low density lipoproteins in blood plasma. To characterize these particles we isolated a cell fraction rich in Golgi apparatus and associated particles from rat liver in quantities sufficient for analysis. Particles freed from the membranes of the Golgi apparatus and floated at d = 1.006 were studied by chemical analysis, immunodiffusion, and paper electrophoresis. The lipid composition of the Golgi particles was similar to that of very low density lipoproteins from the same rats. The protein content was about 10% of dry weight for both the Golgi particles and plasma very low density lipoproteins. The Golgi particles formed lines of identity with plasma very low density lipoproteins during immunodiffusion against antiserum to plasma very low density lipoproteins. On paper electrophoresis, however, many Golgi particles remained near the origin, with only a few migrating to the pre-beta position. It was concluded that the lipoproteins in the Golgi apparatus are the precursors of plasma very low density lipoproteins.     

Subcellular localization of alpha-melanocyte-stimulating hormone in the rat hypothalamus.

Homogenates of male rat hypothalami were fractionated by means of differential centrifugation, and α-melanocyte-stimulating hormone (α-MSH) in the various fractions was quantified by radioimmunoassay. Of the total quantity of α-MSH in the homogenate, 36% was recovered in the 11,500 g pellet and 31% sedimented between 11,500 and 105,000 g. α-MSH was not detected in the 105,000 g supernatant fluid. When the 900 g supernatant fluid was fractionated on continuous sucrose density gradients at non-equilibrium conditions, two populations of particles containing α-MSH were observed. When fractionated at equilibrium conditions, the two populations were recovered in a single band. These sedimentation characteristics indicate that the particles that contain α-MSH differ in size but are similar in density. After hypo-osmotic shock, the large particles containing α-MSH were not demonstrable, whereas the small particles appeared to be resistant to such treatment. In their sedimentation, the particles containing α-MSH were indistinguishable from particles containing thyrotropin releasing hormone (TRH) but were separable from those that contained luteinizing hormone releasing hormone (LHRH). It is suggested that the large particles containing α-MSH are synaptosomes.     

Radial electric field and rotation of the ensemble of plasma particles in tokamak

The velocity of macroscopic rotation of an ensemble of charged particles in a tokamak in the presence of an electric field has been calculated in a collisionless approximation. It is shown that the velocity of toroidal rotation does not reduce to a local velocity of electric drift and has opposite directions on the inner and outer sides of the torus. This result is supplemented by an analysis of the trajectories of motion of individual particles in the ensemble, which shows that the passing and trapped particles of the ensemble acquire in the electric field, on the average, different toroidal velocities. For the trapped particles, this velocity is equal to that of electric drift in the poloidal magnetic field, while the velocity of passing particles is significantly different. It is shown that, although the electric-field-induced shift of the boundaries between trapped and passing particles in the phase space depends on the particle mass and charge and is, in the general case, asymmetric, this does not lead to current generation.     

Changes in the immunogold electron-microscopic localization of calpain in bovine skeletal muscle induced by conditioning and high-pressure treatment

Changes in the localization of calpain in conditioned and pressurized bovine skeletal muscles were investigated by immunogold electron-microscopy. In the muscle immediately after thawing (control), the relative distributions of colloidal particles statistically calculated by counting the colloidal particles were about 65% and 35% in the I-band/Z-disk and A-band regions, respectively. In the muscle conditioned for 7 days, distribution of colloidal particles was more than two times greater in both the I-band/Z-disk and A-band regions than in the control muscle. Almost no change in either the absolute concentration or relative distribution of the colloidal particles was detectable during further storage. In the muscle exposed to a pressure of 100 MPa or 200 MPa, slightly more immunogold was detected in the sarcomere than in that of the control muscle. Increasing pressure up to 300 MPa enabled high-density particles to be seen throughout the sarcomere. Conversely, few particles were detected in the sarcomere of the muscle exposed to 400 MPa. These electron-microscopic observations were confirmed from the statistical analysis as with the conditioned muscles.It was clear from the results obtained that the pressure-induced changes in calpain localization were much more pronounced than those from conditioning.     

Formation of biologically active substances during cholesterol oxidation on the surface of fluorocarbon emulsions

Oxidative modification of cholesterol on the surface of fluorocarbon emulsions was studied. The oxidation yielded one primary product--7-peroxycholesterol. It was shown that the obtained cholesterol C7 derivatives possess a high biological activity. It was concluded that the possibility of oxidative modification of plasma substances on the surface of fluorocarbon emulsion particles with the formation of highly active compounds must be taken into account when using the fluorocarbon particles in medicine.     

A study of the in vitro and in vivo interaction of barium sulphate particles with gastro-intestinal mucus.

The in vivo interaction of various radiopaques with mucus was investigated by a determination of the pH-mobility curves of the treated particles. It was found that mucus covered the radiopaque particles both in vivo and in vitro. This interaction was not affected by the nature of the radiopaque or by the state of the mucous membrane. The changed state of the mucus was modified by the diet of the rat from which it was derived. The results are discussed with reference to the technique used in the administration of radiopaques.     

Inhibition of lipid peroxidation by ubiquinol in submitochondrial particles in the absence of vitamin E

The relationship between the antioxidant effects of reduced coenzyme Q10 (ubiquinol, UQH2) and vitamin E (alpha-tocopherol) was investigated in beef heart submitochondrial particles in which lipid peroxidation was initiated by incubation with ascorbate + ADP-Fe3+. These effects were examined after extraction of coenzyme Q10 (UQ-10) and vitamin E from the particles and reincorporation of the same components alone or in combination. The results show that UQH2 efficiently inhibits lipid peroxidation even when vitamin E is absent. It is concluded that UQH2 can inhibit lipid peroxidation directly, without the mediation of vitamin E.     

Flocculant interaction with native antibiotic solutions. An examination of the mechanism of the process

Some characteristics of the fermentation broth filtrates, i.e. concentration of the disperse particles and some of their parameters (projected length, area, distribution of the particles along the maximum chord) were determined and estimated after flocculation. It was shown that the floccules were mainly formed as fine colloid admixtures present not only in "turbid", but also in visually transparant solutions. It was found that the mechanism of flocculation was rather complicated and not always similar for different batches of the fermentation broth filtrate: it was, in particular, affected by the electrostatic interactions, the flocculant interaction with inorganic reagents, bridge-forming processes.     

Alignment of a restriction map with the genetic map of bacteriophage T4.

We reported previously that polycytidylate [poly(C)]-dependent RNA polymerase activity was a property of small spherical or triangular reovirus-specific particles which sedimented at 13 to 19S and were composed solely of the reovirus protein, sigma NS. Depending on the fraction of cellular extracts from which they were obtained, these particles exhibited marked differences in stability. Most 13 to 19S particles from a particular fraction repeatedly disaggregated into smaller 4 to 5S subunits with no enzymatic activity. Disruption of many particles could be prevented and polymerase activity retained after these particles had bound different single-stranded (ss) RNAs. Our previous results indicated that there was heterogeneity among the 13 to 19S particles in that possession of poly(C)-dependent RNA polymerase activity was a property of only some. Support for this heterogeneity was derived from the demonstration in this report that there were at least three types of binding sites present within particles in any purified preparation: (i) those binding only poly(C); (ii) those binding only reovirus ss RNAs; and (iii) those binding one or the other, but not both at the same time. It is suggested that only those particles able to bind either poly(C) or reovirus ss RNAs had poly(C)-dependent RNA polymerase activity, as reovirus ss RNAs markedly inhibited the polymerase activity. All three size classes of reovirus ss RNAs were equally effective in binding, but once bound, they were not copied. It is possible that heterogeneity in binding capacity of different particles comprised of only one protein, sigma NS, could result from the ability of subunits containing this protein to assemble into slightly different 13 to 19S particles with specificity of binding or polymerase activity conferred by the configuration of the assembled particles. The high capacity of sigma NS to bind many different nucleic acids with some specificity suggests that these particles may act during infection as condensing agents to bring together 10 reovirus ss RNA templates in preparation for double-stranded RNA synthesis.     

Motility in Echinosphaerium nucleofilum. I. An analysis of particle motions in the axopodia and a direct test of the involvement of the axoneme

The motion of particles in the axopodia of Echinosphaerium nucleofilum is saltatory. In the present study, photokymograph records of 123 motions from six axopodia have been analyzed. Particles followed rectilinear paths of from 1 to 15 mum while in continuous motion at an average velocity of 0.66 plus or minus 0.32 mum/s. The velocity of the particles was variable in 36% of the cases measured. Some motions were punctuated by pauses either before continuing in the same direction or reversing. Frequently, several particles were moving at the same velocity, but neighboring particles showed no motion or moved in the opposite direction. Two particles occasionally contacted one another and travelled as a unit for varying lengths of time but subsequently moved independently. These motions reflect the underlying mechanism of motive force production. Furthermore, a glass microneedle can be substituted for the microtubular axoneme in the axopodia. In these artificial axopodia, bidirectional particle motions occurred which were similar to those in normal axopodia. Colchicine, at the threshold dose for axonemal dissolution, had no affect on these particel motions. It is concluded that the microtubular axoneme is not responsible for particle motions and also that individual microtubules are unlikely candidates for motive force production in this system.     

Observations on transmembrane structures of surface immunoglobulin in the plasma membrane of B lymphocytes

We have investigated the possible role of intramembraneous particles as revealed by freeze-fracture electron microscopy in the plasma membrane of B lymphocytes from rabbits and mice as reflections of transmembrane structures of surface immunoglobulin receptor molecules. This was achieved by aggregation of the surface receptors using fluorochrome-conjugated antibodies, fixation and freezing of the cells in 35% glycerol. This procedure resulted in replicas of lymphocytes with well-preserved morphology (no ice-crystals), enabling the study of both protoplasmic and external fracture face in combination with surface receptor markers. It appeared that very small intramembraneous particles (3–6 nm diameter) were selectively clustered under patches of surface receptor label. This phenomenon was found on the external fracture face exclusively and not on the protoplasmic fracture face. ‘Classical’ intramembraneous particles (6–12 nm diameter) were not involved. We suggest that these small, clustered particles should be interpreted as transmembrane structures of surface immunoglobulin molecules.