Binding of β-endorphin and its fragments to the nonopiod receptor of murine peritoneal macrophages

The tritium-labeled selective agonist of the nonopioid β-endorphin receptor the decapeptide immunorphin ([³H]SLTCLVKGFY) with a specific activity of 24 Ci/mmol was prepared. It was shown that [³H]immunorphin binds with a high affinity to the non-opioid β-endorphin receptor of mouse peritoneal macrophages (K _d 2.4 ± 0.1 nM). The specific binding of [³H]immunorphin to macrophages was inhibited by unlabeled β-endorphin (K _i of the [³H]immunorphin-receptor complex 2.9 ± 0.2 nM) and was not inhibited by unlabeled naloxone, α-endorphin, γ-endorphin, and [Met⁵]enkephalin (K _i > 10 μM). Thirty fragments of β-endorphin were synthesized, and their ability to inhibit the specific binding of [³H]immunorphin to macrophages was studied. It was found that the shortest peptide having practically the same inhibitory activity as β-endorphin is its fragment 12–19 (K _i 3.1 ± 0.3 nM).     

Synthetic peptide KKRR corresponding to the human ACTH fragment 15-18 is an antagonist of the ACTH receptor

Tritium-labeled synthetic fragments of human adrenocorticotropic hormone (ACTH) [3H]ACTH (11-24) and [3H]ACTH (15-18) with a specific activity of 22 and 26 Ci/mmol, respectively, were obtained. It was found that [3H]ACTH (11-24) binds to membranes of the rat adrenal cortex with high affinity and high specificity (Kd 1.8 +/- 0.1 nM). Twenty nine fragments of ACTH (11-24) were synthesized, and their ability to inhibit the specific binding of [3H]ACTH (11-24) to adrenocortical membranes was investigated. The shortest active peptide was found to be an ACTH fragment (15-18) (KKRR) (Ki 2.3 +/- 0.2 nM), whose [3H] labeled derivative binds to rat adrenocortical membranes (Kd 2.1 +/- 0.1 nM) with a high affinity. The specific binding of [3H]ACTH-(15-18) was inhibited by 100% by unlabeled ACTH (11-24) (Ki 2.0 +/- 0.1 nM). ACTH (15-18) in the concentration range of 1-1000 nM did not affect the adenylate cyclase activity of adrenocortical membranes and, therefore, is an antagonist of the ACTH receptor.     

Binding of beta-endorphin and its fragments to the non-opiod receptor of murine peritoneal macrophages

The tritium-labeled selective agonist of the nonopioid beta-endorphin receptor the decapeptide immunorphin ([3H]SLTCLVKGFY) with a specific activity of 24 Ci/mmol was prepared. It was shown that [3H]immunorphin binds with a high affinity to the non-opioid beta-endorphin receptor of mouse peritoneal macrophages (Kd 2.4 +/- 0.1 nM). The specific binding of [3H]immunorphin to macrophages was inhibited by unlabeled beta-endorphin (Ki of the [3H]immunorphin-receptor complex 2.9 +/- 0.2 nM) and was not inhibited by unlabeled naloxone, alpha-endorphin, gamma-endorphin, and [Met5]enkephalin (Ki > 10 microM). Thirty fragments of beta-endorphin were synthesized, and their ability to inhibit the specific binding of [3H]immunorphin to macrophages was studied. It was found that the shortest peptide having practically the same inhibitory activity as beta-endorphin is its fragment 12-19 (Ki 3.1 +/- 0.3 nM).     

An ACTH-Like Peptide Immunocortin: Effect on the Activity of Cells from the Rat Adrenal Cortex

The effect of immunocortin, an ACTH-like decapeptide VKKPGSSVKV corresponding to the 11–20 sequence of the variable part of the human IgG1 heavy chain on the content of 11-hydroxycorticosteroids (CS) in rat adrenal glands and blood serum in vivo was studied. An intramuscular injection of immunocortin at a dose of 10 g/kg was found in an hour to induce a twofold decrease in CS content in the adrenal glands and a 1.8-fold increase in the blood serum CS content. At the same time, an immunocortin dose of 100 g/kg exerted practically no effect on the CS content and its dose of 1000 g/kg increased the CS content both in adrenal glands and in blood serum by 1.6 and 2.2 times, respectively. Four hours after the injection of any of the three doses of immunocortin, the CS content in adrenal glands did not differ from the control value, and after 24 h the content decreased threefold. Immunocortin was shown to be bound by the ACTH receptors in the membranes of the rat adrenal cortex with a high affinity and specificity (inhibiting the specific binding of ¹²⁵I-labeled ACTH-(11–24) peptide with K _i of 1.2 nM).     

Interaction of Synthetic Dipeptide Bestim with Mouse Thymocytes and Macrophages

Tritium-labeled dipeptide bestim (γ-D-Glu-L-Trp) with a specific activity of 45 Ci/mmol was obtained by the high-temperature solid-state catalytic isotope exchange (HSCIE) reaction. [³H]bestim was found to bind with high affinity to mouse peritoneal macrophages (K _d 2.1 ± 0.1 nM) and thymocytes (K _d 3.1 ± 0.2 nM) and also plasma membranes isolated from these cells (K _d 18.6 ± 0.2 and 16.7 ± 0.3 nM respectively). The specific bonding of [³H]bestim with macrophages and thymocytes was inhibited by unlabeled dipeptide thymogen (L-Glu-L-Trp) (K _i 0.9 ± 0.1 and 1.1 ± 0.1 nM respectively). Treatment of the macrophages and thymocytes with trypsin led to their loss of capacity to bind [³H]bestim. Bestim at concentrations range of 0.1–1000 nМ reduced the adenylate cyclase activity in macrophage and thymocyte membranes.     

Simulation of the competition for light in forest stands of the same age

A model simulating the growth and interaction of trees during their competition for light has been proposed and verified. Qualitative analysis of the model behavior at various values of parameters determining the tree growth rate has been performed. The model verification shows a sufficiently high accuracy of the model approximation of empirical tree growth data. To study interspecific interactions in forest communities, some computing experiments on the development of mixed forest stands, represented by the main forest-forming wood species of the Far East, have been carried out.     

Self-similar properties of the spatial structure of intertidal macro- and microbenthic communities

Spatial distribution (SD) of White Sea intertidal soft-bottom communities was studied at scales from decimetres to dozens of kilometres on the basis of an extensive dataset (464 samples of macrofauna, 349 samples of ciliates, and 333 samples of diatoms). We used the information index of structural heterogeneity D(I) (Azovsky et al., 2000 // Mar. Biol. 136 (3): 581-590) to characterize spatial variability in the species composition of the communities at different extent (total area surveyed) and grain (finest spatial resolution). The type of distribution was determined via the relation between D(I) and parameters of the spatial scale (extent and grain). At small scale (in terms of extent), all the communities were distributed randomly (mosaic SD). At larger scales, the estimated spatial variability depended neither on extent nor grain, exclusively on their ratio, i.e., was scale-invariant. This means that at some scale the spatial patterns of communities display self-similar properties (fractal SD). Such SD was found at a rather wide range of scales scales: 10(1)-10(4) m for the macrofauna, 10(0)-10(3) m for the ciliates, and 10(-1)-10(2) m for the diatoms. At still greater scales, patchy or gradient patters were observed. Thus, the ranges of fractal distribution were proportional to the average size of the organisms (approximately 10(4)-10(7) times the body size). We suppose that such spatial pattern reflects community self-organization in a relatively homogeneous environment and may be the most efficient way to realize the highest structural diversity on the basis of pre-formed complexes of predominant species. We also suppose that fractal-like patterns may be a general feature of the spatial organization of communities.     

Annual cyclic changes and self-organization processes in the marine sea-bottom organisms

The dynamics of marine microbenthos species structure shows clear annual cycle. From late winter till early autumn community changes towards increasing complexity and orderliness. The sharp increase in available energy (light and heat) in the beginning of the winter acts as trigger of these processes. The further development of the community is connected with system fluctuations arising as a result of interactions or organisms between each other and their environment (feeding, competition, predation). In the end of summer these processes are terminated by the state of high species diversity, maximum species coordination, expansion of inhabited zone within the sediments, and the distinct segregation of space and nutrient resources among species (i.e., achievement of ecological complementarity). Decrease in light and temperature causes the reverse process--weakening of organism coordination and significant simplification of the community structure. In general, these changes correspond to the theory of self-organization in nonequilibrium systems (Prigogine, Stengers, 2001). The most important distinction of observed processes from the classical self-organization is their cyclic dynamics, i.e. the annual return of community to its most simple state at the end of winter.