5.3 S RNA is a discrete cleavage product from the 5′-terminus of 18 S RNA of rat liver ribosomes

A 5.3 S RNA species observed in urea-gel electrophoretic analysis of the RNA of the small ribosomal subunit of rat liver has been identified from its sequence as the 5′-terminal 133–134 base fragment of 18 S RNA. Presumably it is cleaved by an endogenous endonuclease when the ribosomal subunits are dissociated, because it usually is not observed in 18 S RNA obtained by direct extraction of cells or tissues.     

Adhesion,phagocytosis and cell surface energy. The binding of fixed human erythrocytes to rat macrophages and polymethylpentene

Fixed human erythrocytes were used as model particles for the study of adhesion and phagocytosis by rat peritoneal macrophages. Erythrocytes were fixed with various concentrations of glutaraldehyde or tannic acid, or were treated with neuraminidase. Adhesion and phagocytosis of these cells were measured. In addition, the surface energy of these erythrocytes and macrophages was estimated by the contact angle technique. Free energies of adhesion, based on the cell surface energies, were correlated with both adhesion and phagocytosis.     

Metallobiochemistry of ultratrace levels of bismuth in the rat II. Interaction of 205+206Bi3+ with tissue,intracellular and molecular components

BackgroundKnowledge on Bi metabolism in laboratory animals refers to studies at “extreme” exposures, i.e. pharmacologically relevant high-doses (mg kg⁻¹ b.w.) in relation to its medical use, or infinitesimal doses (pg kg⁻¹b.w.) concerning radiobiology protection and radiotherapeutic purposes. There are no specific studies on metabolic patterns of environmental exposure doses (ultratrace level, μg kg⁻¹ b.w.), becoming in this context Bi a “heavy metal fallen into oblivion”. We previously reported the results of the metabolic fate of ultratrace levels of Bi in the blood of rats [1]. In reference to the same study here we report the results of the retention and tissue binding of Bi with intracellular and molecular components.MethodsAnimals were intraperitoneally injected with 0.8 μg Bi kg⁻¹ b.w. as ^205+206Bi(NO)₃, alone or in combination with ⁵⁹Fe for the radiolabeling of iron proteins. The use of ^205+206Bi radiotracer allowed the determination of Bi down to pg fg⁻¹ in biological fluids, tissues, subcellular fractions, and biochemical components isolated by differential centrifugation, size exclusion chromatography, solvent extraction, precipitation, immunoprecipitation and dialysis.Main findingsAt 24 h post injection the kidney contained by far the highest Bi concentration (10 ng g⁻¹ wt.w.) followed by the thymus, spleen, liver, thyroid, trachea, femur, lung, adrenal gland, stomach, duodenum and pancreas (0.1 to 1.3 ng g⁻¹ wt.w.). Brain and testis showed smaller but consistently significant concentrations of the element (0.03 ng g⁻¹ wt.w). Urine was the predominant route of excretion. Intracellularly, liver, kidney, spleen, testis, and brain cytosols displayed the highest percentages (35%–58%) of Bi of homogenates. Liver and testis nuclei were the organelles with the highest Bi content (24 % and 27 %). However, when the recovered Bi of the liver was recorded as percent of total recovered Bi divided by percent of total recovered protein the lysosomes showed the highest relative specific activity than in other fractions. In the brain subcellular fractions Bi was incorporated by neuro-structures with the protein and not lipidic fraction of the myelin retaining 18 % of Bi of the total homogenate. After the liver intra-subcellular fractionation: (i) 65 % of the nuclear Bi was associated with the protein fraction of the nuclear membranes and 35 % with the bulk chromatin bound to non-histone and DNA fractions; (ii) about 50 % of the mitochondrial Bi was associated with inner and outer membranes being the other half recovered in the intramitochondrial matrix; (iii) in microsomes Bi showed a high affinity (close to 90 %) for the membranous components (rough and smooth membranes); (iv) In the liver cytosol three pools of Bi-binding proteins (molecular size > 300 kDa, 70 kDa and 10 kDa) were observed with ferritin and metallothionein–like protein identified as Bi-binding biomolecules. Three similar protein pools were also observed in the kidney cytosol. However, the amount of Bi, calculated in percent of the total cytosolic Bi, were significantly different compared to the corresponding pools of the liver cytosol.ConclusionsAt the best of our knowledge the present paper represents the first in vivo study, on the basis of an environmental toxicology approach, aiming at describing retention and binding of Bi in the rat at tissue, intracellular and molecular levels.     

The effect of phenobarbital and 3′-methyl-N,N-dimethyl-4-aminoazobenzene administration on catalytic,binding and immunological properties of ligandin subunits in rat liver

Following administration of phenobarbital to rats, liver ligandin content, bilirubin binding, glutathione-S-transferase and steriod isomerase activities by 150% and the 22 000-dalton subunit was selectively increased. Following adminstration of 3′-methyl-N,N-dimethyl-4-aminoazobenzene, rat liver ligandin content and steroid isomerase decrased by 65%, glutathione-S-transferase incrased by 100%, bilirubin binding was abolished, and the relative proportion of the 22 000- and 25 000-dalton subunits remained unchanged.     

Multi-scale modeling of complex neuronal networks: a view towards striatal cholinergic pattern formations

The phenomena related to brain function occur as the interplay of various modules at different spatial and temporal scales. Particularly, the integration of the dynamical behavior of cells within the complex brain topology reveals a heterogeneous multi-scale problem, which has, to date, mainly been addressed by methods of statistical physics such as mean-field approximations. In contrast, the present study introduces an abstract mathematical model of a deterministic nature that provides a robust integral transformation of the microscopic activities into macroscopic spatiotemporal patterns. The existence of the transformation operator is guaranteed by the convergence of a repetitive patching of the network domain with its fundamental domains that express the local topologies of the tissue. Depending on the choice of the local connectivity function, this framework represents a computationally efficient generalization of the classical Kirchhoff’s, Hebbian, and Hopfield’s approaches. The capabilities of this multi-scale method have been evaluated within the structure of the dorsal striatum of rats, a brain region with major involvement in motor and cognitive information processing. Numerical simulations suggest the formation of characteristic spatiotemporal patterns due to the activation of cholinergic interneurons.     

Nippostrongylus brasiliensis: Peripheral blood leucocyte response of rats,with special reference to basophils

Changes in peripheral blood leucocytes were followed in male August rats given one or two infections with the parasitic nematode, Nippostrongylus brasiliensis. During the initial infection, there was a biphasic increase in total numbers of leucocytes, lymphocytes, neutrophils, large mononuclear cells, and eosinophils. All except eosinophils fell rapidly to normal levels as the parasites were expelled, but eosinophils were elevated much longer. All these cell types increased in number to a single peak 5 days after reinfection. Basophils were detected at very low levels in uninfected rats (0.06% or $\text{1}\text{1600}$ leucocytes) and increased in number to a peak 13 days after initial infection, at which time they represented about 4.5% of total leucocytes, an 80-fold increase compared with the number in normal rats. In reinfected rats, the basophilia occurred more rapidly than in a primary infection, suggesting that the appearance of these cells in the circulation is probably an immunologically mediated event.     

The involvement of parenchymal,Kupffer and endothelial liver cells in the hepatic uptake of intravenously injected liposomes effects of lanthanum and gadolinium salts

¹²⁵I-labeled albumin or poly(vinyl pyrrolidone) encapsulated in intermediate size multilamellar or unilamellar liposomes with 30–40% of cholesterol were injected intravenously into rats. In other experiments liposomes containing phosphatidyl[Me-¹⁴C]choline were injected. 1 h after injection parenchymal or non-parenchymal cells were isolated. Non-parenchymal cells were separated by elutriation centrifugation into a Kupffer cell fraction and an endothelial cell fraction. From the measurements of radioactivities in the various cell fractions it was concluded that the liposomes are almost exclusively taken up by the Kupffer cells. Endothelial cells did not contribute at all and hepatocytes only to a very low extent to total hepatic uptake of the ¹²⁵I-labels. Of the ¹⁴C-label, which orginates from the phosphatidylcholine moiety of the liposomes, much larger proportions were recovered in the hepatocytes. A time-dependence study suggested that besides the involvement of phosphatidylcholine exchange between liposomes and high density lipoprotein, a process of intercellular transfer of lipid label from Kupffer cells to the hepatocytes may be involved in this phenomenon. Lanthanum or gadolinium salts, which effectively block Kupffer cell activity, failed to accomplish an increase in the fraction of liposomal material recovered in the parenchymal cells. This is compatible with the notion that liposomes of the type used in these experiments have no, or at most very limited, access to the liver parenchyma following their intravenous administration to rats.     

Myometrial connexin 43 trafficking and gap junction assembly at term and in preterm labor.

Modulation of connexin 43 (cx43) in the myometrium of timed pregnant rats was studied using enzyme-linked immunosorbent assay (ELISA), immunocytochemical localization, and immunoblot. These techniques utilized site-specific antibodies directed against a portion of the carboxyl tail of cx43. We found that cx43 is synthesized several days prior to labor but accumulates within the cytoplasm until parturition, when it is rapidly transported to the plasma membrane and assembled into gap junction plaques at the cell surface. These cx43-positive gap junctions begin to disappear from the plasma membrane within hours of delivery of the last pup and are completely absent within 24 hr following delivery. These structures are apparently internalized and degraded within the cytoplasm. ELISA documents a reduction of total cellular cx43 to baseline levels within 5 days following parturition. While the timing of synthesis, cytoplasmic storage, concentration in apparent Golgi vesicles, and transport to and assembly in the plasma membrane are accelerated in three models of preterm labor, the sequence of these events and the correlation of parturition with the formation of gap junctions are identical to those documented in normal labor. These results support the hypothesis that effective labor requires the synthesis and assembly of cx43 into functional gap junctions at the myometrial cell surface.     

Dietary supplementation of N-3 fatty acids and hydroperoxide levels in rat retinas

Docosahexaenoic acid (DHA) plays an important role in visual and neural development in mammals. In the present study, effect of dietary supplementation with n-3 fatty acids, primarily docosahexaenoic acid (DHA) with high purity, on the fatty acid composition of photoreceptor cells of young rats (fed from 4 weeks) was investigated. DHA in rod outer segment (ROS) membranes was significantly increased in the group of high DHA feeding (9.69% total energy). Other n-3 fatty acids (α-linolenic acid (ALA) and eicosapentaenoic acid (EPA)) included in the diets with DHA (0.95%～5.63% total energy) also significantly increased the proportion of DHA compared with the linoleic acid diet groups. However, the proportions of arachidonic acid (ARA) and other long chain n-6 fatty acids (22:4n6 and 22:5n6) were suppressed in these n-3 fatty acids-fed groups. Phospholipid hydroperoxides in ROS membranes were determined using a highly sensitive analytical technique, chemiluminescence-high performance liquid chromatography (CL-HPLC). There was no increasing tendency in the hydroperoxide levels of ROS membranes containing high content of DHA, and phosphatidylethanolamine hydroperoxide (PEOOH) was much lower than phosphatidylcholine hydroperoxide (PCOOH) under normal light conditions, which implies that DHA supplementation does not much affect the peroxidizability of ROS membranes in vivo. But UV irradiation on separated ROS membranes accelerated the formation of phospholipid hydroperoxides in high DHA feeding rats, and PEOOH was produced more efficiently than PCOOH in vitro.     

Intracellular Ca2+ levels in rat ventricle cells exposed to extremely low frequency magnetic field

Objective: Electromagnetic fields can affect intracellular Ca²⁺ levels. The aim of this study was to determine the changes intracellular Ca²⁺ concentration in cardiac ventricle cells of rats exposed to 0.25 mT (2.5 Gauss) magnetic field.

Methods: Forty-five male rats were introduced to this study. The rats were divided into three groups: control, sham, and experiment. The experimental group was exposed to 0.25 mT extremely low frequency (ELF) magnetic field for 14 days, 3 h/day. The sham group was treated like the experimental group, except for elf-magnetic field exposure. The control group was not subjected to anything and differed from the experimental group and sham group. In the end of experiment, rats were sacrificed, cardiac tissue was removed, and these were fixed in 10% neutral formalin. Then, ventricular cells were stained by Alizarin red staining method.

Results: In the light microscopic examinations of control groups, in myofibril structures between groups, changes were not observed. In myofibril regions of the experimental group compared to other groups, increased heterogen Ca²⁺ accumulations were found.

Conclusion: ELF magnetic fields are used in daily life. The results of this study show that intracellular Ca²⁺ accumulation in cardiac ventricles can increase in rats exposed to ELF magnetic field.