Comparison of the physiological and biochemical indices of the phagocytosis of ram erythrocytes by macro- and microphages]

Morita and Perkins' method was applied to the study of the stage of ingestion and destruction of an antigen (sheep erythrocytes) in the macrophages of peritoneal exudate of rabbits and rats and in the microphages of rabbit pleural exudate. Ingestion and intracellular destruction of the antigen were accompanied by intensified respiration and glycolysis of phagocytes. Respiration of the three types of phagocytes at two stages of phagocytosis and also the digestive capacity of microphages proved to be sensitive to cyanide and colchicine. The latter failed to influence the ingestion of the antigen by the three types of phagocytes and its digestion by macrophages. The differences in the metabolism routes of macro- and microphages in intracellular destruction of the antigen was postulated. An intensification of the phagocytic activity after the immunization was characteristic of rabbit and rat macrophages.     

Complex surveillance of Streptococcus pyogenes V. Protective features complex surveillance of streptococcus of cell-mediated immunity against group a streptococci

Exoproducts of incubated white blood elements from persons hypersensitive to zymosan were found to promote the phagocytic activity of human microphages (and guinea pig macrophages) under conditions of a combined model of "MIF-opsonophagocytic test". The stimulation resulted in a bacteriostatic effect, was non-type-specific and improved microphage potential to inactivate streptococci opsonized with anti-M antibodies in low titre. The microphages pre-exposed to anti-M sera remained unaffected, the effect was non-species-specific and nontype-specific and the stimulation resulted from the action of albumin fraction exoproducts. This seems to suggest that human mediators (lymphokines) may promote the activity of phagocytes. The cell-mediated immunity appears thus to have protective features even in the case of streptococci which are the typical extracellular parasites. The drawbacks of the model used, which make it difficult to quantify the found mechanism under in vivo conditions, are discussed in detail.     

Diversity and abundance of single-stranded DNA viruses in human feces

In this study, we investigated the abundance and diversity of single-stranded DNA (ssDNA) viruses in fecal samples from five healthy individuals through a combination of serial filtration and CsCl gradient ultracentrifugation. Virus abundance ranged from 10⁸ to 10⁹ per gram of feces, and virus-to-bacterium ratios were much lower (less than 0.1) than those observed in aquatic environments (5 to 10). Viral DNA was extracted and randomly amplified using phi29 polymerase and analyzed through high-throughput 454 pyrosequencing. Among 400,133 sequences, an average of 86.2% viromes were previously uncharacterized in public databases. Among previously known viruses, double-stranded DNA podophages (52 to 74%), siphophages (11 to 30%), myophages (1 to 4%), and ssDNA microphages (3 to 9%) were major constituents of human fecal viromes. A phylogenetic analysis of 24 large contigs of microphages based on conserved capsid protein sequences revealed five distinct newly discovered evolutionary microphage groups that were distantly related to previously known microphages. Moreover, putative capsid protein sequences of five contigs were closely related to prophage-like sequences in the genomes of three Bacteroides and three Prevotella strains, suggesting that Bacteroides and Prevotella are the sources of infecting microphages in their hosts.     

Changes in the size and density of human high-density lipoproteins promoted by a plasma-conversion factor

Incubation of human high-density lipoprotein subfraction-3 (HDL3) with rabbit lipoprotein-depleted plasma resulted in marked changes in the density and size of the HDL. After 24 h of incubation at 37 degrees C, the original HDL3 were converted into populations of larger (less dense) and smaller (more dense) particles. The degree of conversion increased with increasing concentrations of lipoprotein-depleted plasma and increasing incubation time. Furthermore, lecithin:cholesterol acyltransferase, lipoprotein lipase and lipid-transfer protein were shown not to be involved in the process. It was therefore proposed that a separate factor, the HDL-conversion factor, was responsible for the observed changes. Conversion-factor activity was assessed in the lipoprotein-depleted plasma of several species and found to be greater in rabbits and rats than in pigs and human subjects. It was also established that the conversion factor was able to be precipitated from rabbit lipoprotein-depleted plasma between 40 and 50% saturation of (NH4)2SO4. This information was used to partially purify the factor from human plasma. The proteins of human plasma which precipitated between 35 and 55% saturation of (NH4)2SO4 were recovered and subjected to ultracentrifugation to isolate the fraction of density 1.21-1.25 g/ml. This fraction, which was rich in HDL-conversion activity, was further purified by cation-exchange chromatography. In conclusion, a factor which promotes the conversion of HDL to populations of larger and smaller particles has been found to exist at various levels of activity in the plasma of several species. Partial purification of the factor from human plasma has been achieved.     

Changes in the Asymmetry of Eeg Waves During Voluntary Word Memorization

Study of electroencephalographic (EEG) correlates of psychological processes is becoming an increasingly popular subject, and scientists are gradually abandoning the notion that EEG parameters contain no useful information about the activity in which the subject is engaged. The search for various EEG indices reflecting the characteristics of the work being performed by a subject is being pursued with increasing vigor. The hypothesis behind these studies is that as a person goes about performing some activity, specific changes in neurodynamics take place in various locations in the cerebral cortex and that these changes will be reflected in the corresponding EEG indices. Furthermore, these EEG changes will persist as long as the subject continues what he is doing, but will be especially intense at critical moments during that activity: during the first stages, at moments preceding or accompanying the discovery of an algorithm for solving the given problem, when wrong responses are given, etc.     

Design and Synthesis of Lipopolysaccharide-Binding Antimicrobial Peptides Based on Truncated Rabbit and Human CAP18 Peptides and Evaluation of Their Action Mechanism

Lipopolysaccharide (LPS) is a toxic and immunogenic agent for human. Additionally, LPS is a good target for some antimicrobial compounds, including antimicrobial peptides (AMPs). LPS-binding peptides (LBPs) can recognize and neutralize LPS. Rabbit and human cathelicidins are AMPs with LPS-binding activity. In this study, we designed and synthesized two new truncated LBPs from rabbit and human CAP18 peptides by in silico methods. After synthesis of peptides, the antimicrobial properties and LPS-binding activity of these peptides were evaluated. The parental rabbit and human CAP18 peptides were selected as positive controls. Next, the changes in the secondary structure of these peptides before and after treatment with LPS were measured by circular dichroism (CD). Human cytotoxicity of the peptides was evaluated by MTT and red blood cells (RBCs) hemolysis assays. Finally, field emission scanning electron microscopy (FE-SEM), confocal microscopy, and flow cytometry were performed to study the action mechanism of these peptides. Results indicated that the hCap18 and rCap18 had antibacterial activity (at a MIC of 4–128 μg/mL). The results of the quantitative LAL test demonstrated that LPS-binding activity of hCap18 peptide was better than rCap18, while rCap18 peptide had better antimicrobial properties. Furthermore, rCap18 had less cytotoxicity than hCap18. However, both peptides were nontoxic for normal human skin fibroblast cell in MIC range. In conclusion, rCap18 has good antibacterial properties, while hCap18 can be tested as a diagnostic molecule in our future studies.

     

Hyaluronic acid is an endogenous inducer of interleukin-1 production by human monocytes and rabbit macrophages

When human peripheral monocytes and rabbit peritoneal macrophages were incubated with hyaluronic acid, the media were found to contain interleukin-1 (IL-1) activity and to stimulate collagenase production by rabbit fibroblasts. A digestion of hyaluronic acid by testicular hyaluronidase decreased the IL-1 inducing activity. Polymixin B, an inhibitor of endotoxin, did not exert any effect towards the action of hyaluronic acid. Hyaluronic acid also stimulated human polymorphonuclear leucocytes to produce IL-1 like activity. These results indicate that hyaluronic acid is an endogenous IL-1 inducer and may play important roles in the pathological and/or physiological changes of connective tissues.     

Correlation of biophysical parameters of biologically active points and variations of heliogeophysical factors

The correlation between the parameters of biologically active points and the dynamics of solar activity was studied. Statistically significant correlations between electroconductivity of biologically active points and solar activity indices (sunspot number, Ap-index, solar radiowave flux) were revealed. The spectral analysis of the temporal organization of biologically active points revealed a set of periods present in the dynamics of solar activity. It was concluded that bioelectric system of the human organism is a complicated multilevel time-structure, which effectively reacts to changes in heliogeophysical dynamics. The bioelectric system of biologically active points is considered to be one of general information channels of electromagnetic nature, which realize the connection of organism with environment.     

Electron microscopical demonstration of acid phosphatase in blood platelets

Summary Ultrastructures of human and rabbit thrombocytes reveal specific subcellular organelles within these elements. Serotonin granules are demonstrated containing extremely electron opaque material in vesicles with an average diameter of 1,700 Å and a considerable number of large dense bodies (average size 4,000 Å in diameter) is seen. The latter are less electron dense as compared to the serotonin granules. The appearance of serotonin granules in the human thrombocyte is rare, while rabbit platelets show a higher number of these granular vesicles.Acid phosphatase activity in the large dense bodies of human and rabbit platelets has been demonstrated by means of electron microscopy. Present results together with currently available biochemical information are briefly discussed in relation to the lysosomal activity within the thrombocytes.     

Refractive indices of the collagen fibrils and extrafibrillar material of the corneal stroma.

Ultrastructural data from x-ray diffraction studies of the cornea were used to estimate the refractive indices of the collagen fibrils and extrafibrillar material of human, ox, trout, and rabbit corneas. X-ray diffraction measurements of the size and spacing of the collagen fibrils and the separation between the constituent molecules of the fibrils were taken from a previous species study. The tissue volume fractions occupied by the stromal components were estimated and their refractive indices were calculated using the Gladstone-Dale law of mixtures. For the fibrils and extrafibrillar material, the refractive indices in the human cornea were 1.411 and 1.365; for the ox 1.413 and 1.357; for the rabbit 1.416 and 1.357; and for the trout 1.418 and 1.364, respectively. An alternative estimate based on the physical properties and chemical composition of bovine cornea, accounting for interfibrillar type VI collagen and cellular water, produced similar estimates of 1.416 and 1.356 for the fibrils and extrafibrillar material, respectively.     

Comparative aspects of transferrin-reticulocyte interactions: membrane receptors and iron uptake

1. A comparative study was made of transferrin and iron uptake by rabbit, rat and human reticulocytes and chick embryo erythrocytes from rabbit, rat, human, chicken and porcine transferrins, human lactoferrin and chicken conalbumin. 2. Three methods were used, viz. direct and competitive uptake studies of transferrin and iron by the four species of cells, and competitive studies of transferrin binding by solubilized membrane receptors (rabbit reticulocytes only). 3. Methods were devised to analyse the data so as to obtain indices of relatedness or relative affinities of each type of heterologous transferrin in rates of iron uptake found with transferrin and cells from various species are largely due to variation in the affinity of cellular receptors for different transferrins. 5. It is concluded that the procedure used in this investigation allow the assessment of phylogenetic relationships and evolutionary trends obtained by structural studies of proteins.     

Isolation and characterization of human nuclear and cytosolic multisynthetase complexes and the intracellular distribution of p43/EMAPII

In this study, the human multienzyme aminoacyl-tRNA synthetase "core" complex has been isolated from the nuclear and cytosolic compartments of human cells and purified to near homogeneity. It is clear from the polypeptide compositions, stoichiometries, and three-dimensional structures that the cytosolic and nuclear particles are very similar to each other and to the particle obtained from rabbit reticulocytes. The most significant difference observed via aminoacylation activity assays and densitometric analysis of electrophoretic band patterns is a lower amount of glutaminyl-tRNA synthetase in the human particles. However, this is not enough to cause major changes in the three-dimensional structures calculated from samples negatively stained with either uranyl acetate or methylamine vanadate. Indeed, the latter samples produce volumes that are highly similar to an initial structure previously calculated from a frozen hydrated sample of the rabbit multisynthetase complex. New structures in this study reveal that the three major structural domains have discrete subsections. This information is an important step toward determination of specific protein interactions and arrangements within the multisynthetase core complex and understanding of the particle's cellular function(s). Finally, gel filtration and immunoblot analysis demonstrate that a major biological role for the cytokine precursor p43 is as an integral part of the multisynthetase complex.     

The Use of Actimetry to Assess Changes to the Rest–Activity Cycle

The endogenous circadian oscillator (the body clock) is slow to adjust to altered rest–activity patterns. As a result, several negative consequences arise during night work and after time‐zone transitions. The process of adjustment can be assessed by measurements of the sleep electroencephalogram (EEG), core temperature or melatonin secretion, for example, but these techniques are very difficult to apply in field studies, and make very great demands upon both experimenters and subjects. We have sought to establish if the activity record, measured conveniently and unobtrusively by a monitor attached to the wrist, can be treated in ways that enable estimates to be made of the disruption caused by changes to the rest–activity cycle, and the process of adjustment to them. In Part A, we describe the calculation and assessment of a series of “activity indices” that measure the overall activity pattern, activity when out of bed or in bed, or the activity in the hours adjacent to going to bed or getting up. The value of the indices was assessed by measuring changes to them in subjects undergoing night work or undergoing time‐zone transitions. In both cases, there is a large body of literature describing the changes that would be expected. First, night workers (working 2 to 4 successive night shifts) were investigated during rest days and night shifts. The indices indicated that night work was associated with lower activity when the subjects were out of bed and higher activity when in bed. Some indices also measured when subjects took an afternoon nap before starting a series of night shifts and gave information about the process of adjustment to night work and recovery from it. Second, in studies from travelers crossing six or more time zones to the east or west, the indices indicated that there were changes to the rest–activity cycle immediately after the flights, both in its overall profile and when activity of the subjects in bed or out of bed was considered, and that adjustment took place on subsequent days. By focusing on those indices describing the activity records during the last hour in bed (LHIB) and the first hour out of bed (FHOB), some evidence was found for incomplete adjustment of the body clock, and for differences between westward and eastward flights. In Part B, the battery of indices are applied to the activity records of long‐haul pilots, whose activity patterns showed a mixture of effects due to night work and time‐zone transitions. Actimetry was performed during the flights themselves and during the layover days (which were either rest or work days). The indices indicated that all pilots had disrupted rest–activity cycles caused by night flights, and that there were added problems for those who had also undergone time‐zone transitions. Rest days were valuable for normalizing the activity profile. For those pilots who flew to the west, adjustment was by delay, though not all aspects of the rest–activity cycle adjusted immediately; for those who flew to the east, some attempted to advance their rest–activity cycle while others maintained home‐based activity profiles. The indices indicated that the activity profile was disrupted more in those pilots who attempted to advance their rest–activity cycle. We conclude that objective estimates of the disruption caused to the rest–activity cycle and the circadian system can be obtained by suitable analysis of the activity record.     

The use of actimetry to assess changes to the rest-activity cycle

The endogenous circadian oscillator (the body clock) is slow to adjust to altered rest-activity patterns. As a result, several negative consequences arise during night work and after time-zone transitions. The process of adjustment can be assessed by measurements of the sleep electroencephalogram (EEG), core temperature or melatonin secretion, for example, but these techniques are very difficult to apply in field studies, and make very great demands upon both experimenters and subjects. We have sought to establish if the activity record, measured conveniently and unobtrusively by a monitor attached to the wrist, can be treated in ways that enable estimates to be made of the disruption caused by changes to the rest-activity cycle, and the process of adjustment to them. In Part A, we describe the calculation and assessment of a series of “activity indices” that measure the overall activity pattern, activity when out of bed or in bed, or the activity in the hours adjacent to going to bed or getting up. The value of the indices was assessed by measuring changes to them in subjects undergoing night work or undergoing time-zone transitions. In both cases, there is a large body of literature describing the changes that would be expected. First, night workers (working 2 to 4 successive night shifts) were investigated during rest days and night shifts. The indices indicated that night work was associated with lower activity when the subjects were out of bed and higher activity when in bed. Some indices also measured when subjects took an afternoon nap before starting a series of night shifts and gave information about the process of adjustment to night work and recovery from it. Second, in studies from travelers crossing six or more time zones to the east or west, the indices indicated that there were changes to the rest-activity cycle immediately after the flights, both in its overall profile and when activity of the subjects in bed or out of bed was considered, and that adjustment took place on subsequent days. By focusing on those indices describing the activity records during the last hour in bed (LHIB) and the first hour out of bed (FHOB), some evidence was found for incomplete adjustment of the body clock, and for differences between westward and eastward flights. In Part B, the battery of indices are applied to the activity records of long-haul pilots, whose activity patterns showed a mixture of effects due to night work and time-zone transitions. Actimetry was performed during the flights themselves and during the layover days (which were either rest or work days). The indices indicated that all pilots had disrupted rest-activity cycles caused by night flights, and that there were added problems for those who had also undergone time-zone transitions. Rest days were valuable for normalizing the activity profile. For those pilots who flew to the west, adjustment was by delay, though not all aspects of the rest-activity cycle adjusted immediately; for those who flew to the east, some attempted to advance their rest-activity cycle while others maintained home-based activity profiles. The indices indicated that the activity profile was disrupted more in those pilots who attempted to advance their rest-activity cycle. We conclude that objective estimates of the disruption caused to the rest-activity cycle and the circadian system can be obtained by suitable analysis of the activity record.     

Modification of bovine somatotropin (growth hormone) with plasmin

Although much work has been reported on modification of human somatotropin with plasmin and has revealed important information about structure-function relationships, plasmin modification of nonprimate somatotropins (which differ markedly in structure and biological actions from the human hormone) has been little studied. Therefore we investigated plasmin digestion of bovine somatotropin. Digestion was less rapid but more extensive than that of human somatotropin. Structural characterization of digestion products resolved by gel filtration suggested that a major product after 24 h was a disulfide-linked fragment comprising residues 1–133 plus 140–177. Further cleavage products were found in aggregated material and minor fractions. In radioimmunoassay for bovine somatotropin, activity was retained only by the unfractionated digest and aggregated material. In radioreceptor assay for somatotropin using receptors from livers of late-pregnant rabbit all preparations retained some activity. The results suggest that receptor- and antibody-binding sites in bovine somatotropin are not identical and that greater activity may result from specific association of fragments that are less active or inactive once separated.     

Comparative analysis of the locus control region of the rabbit beta-like gene cluster: HS3 increases transient expression of an embryonic epsilon-globin gene.

The rabbit homolog to the locus control region (LCR) of the human beta-like globin gene cluster was isolated, and long segments containing the DNase I hypersensitive sites (HS) were sequenced. The order and spacing of HS4, HS3, HS2 and HS1 are conserved between rabbit and human. Alignment of these sequences with their homologs from human, goat, and mouse shows that very long segments of DNA match between species, for over a thousand base pairs on either side of the previously identified functional cores, indicating that some important functions are found outside the cores. The activity of rabbit HS2 and HS3 was tested by attaching each to a novel reporter gene constructed by inserting the luciferase coding region into the rabbit epsilon-globin gene. In contrast to previous reports showing no effect of human or mouse HS3 on transient expression, both the rabbit HS2 and HS3 DNA fragments separately increased transient expression from the epsilon-luciferase hybrid gene and expression from stably integrated constructs in K562 erythroleukemia cells.     

Molecular stability of chicken and rabbit immunoglobulin G.

Molecular stability of chicken egg yolk immunoglobulin G (IgY) and that of rabbit IgG were compared by measuring antibody activities and conformational changes. Stability of rabbit IgG to acid denaturation was much higher than that of IgY. Conformation of the IgY molecule was readily changed in acidic conditions, resulting in a rapid loss of antibody activity. Much less stable natures of IgY to heat-treatment and guanidine-HCl denaturation than rabbit IgG were also observed. Differences in the structure between the two immunoglobulins that might participate in their different stability were inferred from their amino acid sequence data. Importance of the intramolecular disulfide linkage in the rabbit light chain and some other structural differences were suggested.     

Modifications in the activities of membrane-bound enzymes during in vivo ageing of human and rabbit erythrocytes

Erythrocyte plasma membranes were isolated from a homogeneous population of human or rabbit erythrocytes fractionated into classes representing young, middle-age and old age in vivo. Lipid analyses of human erythrocyte plasma membranes reveal a decrease of the cholesterol to phospholipid molar ratio, followed by a marked decrease in the activities of the membrane-bound enzymes (Na+,K+)-stimulated ATPase, acetylcholinesterase and NAD+ase from young to old age. Such changes were not observed between young and middle-age rabbit erythrocytes. Incubation of rabbit young erythrocytes with phosphatidylcholine vesicles (liposomes) to obtain partial depletion of their membrane cholesterol, indicated that cholesterol depletion causes a statistically significant decrease of the (Na+,K+)-stimulated ATPase and acetylcholinesterase activities, but the NAD+ase activity remained almost unchanged. The biological significance of these data are discussed in terms of the differences and modifications in the interaction of membrane-bound enzymes with membrane lipids during in vivo ageing of erythrocytes.     

The biological effects in animals in relation to the accident at the Chernobyl Atomic Electric Power Station. 7. The content of lipid peroxidation products and free fatty acids and the catalase activity in a number of organs and tissues of rats]

The content of malonic dialdehyde, dienic conjugates and free fatty acids, and catalase activity were determined in some organs and tissues of chronically irradiated rats. Certain regularities were found in lipid metabolism changes. The MIGI-K preparation was shown to normalize some indices under study.