Cell specific quantitative iron mapping on brain slices by immuno-µPIXE in healthy elderly and Parkinson's disease.

Iron is essential for neurons and glial cells, playing key roles in neurotransmitter synthesis, energy production and myelination. In contrast, high concentrations of free iron can be detrimental and contribute to neurodegeneration, through promotion of oxidative stress. Particularly in Parkinson's disease (PD) changes in iron concentrations in the substantia nigra (SN) was suggested to play a key role in degeneration of dopaminergic neurons in nigrosome 1. However, the cellular iron pathways and the mechanisms of the pathogenic role of iron in PD are not well understood, mainly due to the lack of quantitative analytical techniques for iron quantification with subcellular resolution. Here, we quantified cellular iron concentrations and subcellular iron distributions in dopaminergic neurons and different types of glial cells in the SN both in brains of PD patients and in non-neurodegenerative control brains (Co). To this end, we combined spatially resolved quantitative element mapping using micro particle induced X-ray emission (µPIXE) with nickel-enhanced immunocytochemical detection of cell type-specific antigens allowing to allocate element-related signals to specific cell types. Distinct patterns of iron accumulation were observed across different cell populations. In the control (Co) SNc, oligodendroglial and astroglial cells hold the highest cellular iron concentration whereas in PD, the iron concentration was increased in most cell types in the substantia nigra except for astroglial cells and ferritin-positive oligodendroglial cells. While iron levels in astroglial cells remain unchanged, ferritin in oligodendroglial cells seems to be depleted by almost half in PD. The highest cellular iron levels in neurons were located in the cytoplasm, which might increase the source of non-chelated Fe3+, implicating a critical increase in the labile iron pool. Indeed, neuromelanin is characterised by a significantly higher loading of iron including most probable the occupancy of low-affinity iron binding sites. Quantitative trace element analysis is essential to characterise iron in oxidative processes in PD. The quantification of iron provides deeper insights into changes of cellular iron levels in PD and may contribute to the research in iron-chelating disease-modifying drugs.

Regulation of the peritoneal macrophage functional activity by the MP-5 and MP-6 myelopeptides under stress.

In mice, two-hour immobilization stress inhibited zymosan-induced production by macrophages of the oxygen radicals and cytokine IL-1ß. After myelopeptides MP-5 and MP-6 were administered into mice, the stress-induced inhibition of the reactive oxygen species (ROS) and IL-1ß was abrogated. MP-5 peptide stimulated spontaneous ROS production by macrophages and reduced IL-10 production under stress. Thus, under in vivo conditions and under stress, the effect of MP-5 and MP-6 myelopeptides modulates the peritoneal macrophage activity.

[Effects of myelopeptide-1 on functional activity of murine phagocytes treated with cyclophosphan].

The effects of bone marrow derived myelopeptide-1 (MP-1) on functional activities of normal murine phagocytes and phagocytes obtained from mice treated with cytostatic agent cyclophosphan (CY) were studied in vivo and in vitro. We demonstrated immunocorrecting effect of MP-1 on functional activity of bone marrow and peripheral blood phagocytes. An optimal protocol of MP-1 injections during cytostatic therapy was developed to obtain maximal efficacy in immunocorrection. The best effect on functional activity of phagocytes of different localization (the highest effect on peripheral blood neutrophils) was found when MP-1 was injected before CY. The results demonstrated that MP-1 can be considered as a substance able to protect peripheral blood phagocytes from CY damage during cytostatic treatment.

[Especially in the hemostatic system in geriatric patients with ischemic heart disease, complicated by persistent atrial fibrillation].

The article presents the results of the state of the vascular-platelet, coagulation hemostasis and fibrinolysis system in the elderly with coronary heart disease complicated with persistent atrial fibrillation.

[Synthesis and anti-tumor properties of myelopeptide MP-1].

We have studied anti-tumor properties of bone marrow derived peptide Phe-Leu-Gly-Phe-Pro-Thr (MP-1) synthesized by classical methods. It was shown that MP-1 enhanced the effect ofcytostatic therapy of lymphatic leukemia P388 and increased latent growth period of P388 tumors implanted in irradiated mice. MP-1 also decreased metastasis of mouse breast adenocarcinoma Ca-755 after surgery.

Current CONtrolled Transmit And Receive Coil Elements (CONTAR) for Parallel Acquisition and Parallel Excitation Techniques at High-Field MRI.

A novel intrinsically decoupled transmit and receive radio-frequency coil element is presented for applications in parallel imaging and parallel excitation techniques in high-field magnetic resonance imaging. Decoupling is achieved by a twofold strategy: during transmission elements are driven by current sources, while during signal reception resonant elements are switched to a high input impedance preamplifier. To avoid B(0) distortions by magnetic impurities or DC currents a resonant transmission line is used to relocate electronic components from the vicinity of the imaged object. The performance of a four-element array for 3 T magnetic resonance tomograph is analyzed by means of simulation, measurements of electromagnetic fields and bench experiments. The feasibility of parallel acquisition and parallel excitation is demonstrated and compared to that of a conventional power source-driven array of equivalent geometry. Due to their intrinsic decoupling the current-controlled elements are ideal basic building blocks for multi-element transmit and receive arrays of flexible geometry.

[Synthesis and properties of the myelopeptides with differentiating activity].

The bone marrow myelopeptides Phe-Arg-Pro-Arg-Ile-Met-Thr-Pro (MP-4) and Val-Asp-Pro-Pro (MP-6) have been synthesised by a classical method and by a solid phase synthesis. The differentiating activity of MP-4 and MP-6 in human leukemia cells HL-60 and K-562 has been studied. Both peptides induce terminal differentiation of these cell lines but the mechanism of action of peptides MP-4 and MP-6 is distinguished.

Myelopeptide-5 abolishes virus-induced immunosuppression by restoring T cell functions.

Myelopeptide-5 (MP-5; synthetic analog of endogenous low-molecular-weight peptide Val-Val-Tyr-Pro-Asp) neutralized the immunosuppressive effects of antiviral vaccines (influenza, measles, and measles/parotitis) and stimulated their immunogenicity by restoring functional activity of T cells, suppressed by the viruses. Specific binding of MP-5 to CD4(+) lymphocyte (its target cell) was studied using [(3)H]MP-5 (dissociation constant 2.03 x 10(-7) M).

[Fluorescently labeled differentiating myelopeptide-4: specific binding to and penetration into target cells].

Myelopeptide-4 (MP-4) (Phe-Arg-Pro-Arg-Ile-Met-Thr-Pro), inducing the terminal differentiation of HL-60 leukemia cells, was labeled with fluorescein isothiocyanate. The specific binding of this modified peptide to the surface of HL-60 cells and its ability to penetrate into the cells were studied. It was shown by cytometry and confocal microscopy to be bound on the HL-60 cell surface, to penetrate into their cytoplasm, and finally to concentrate around the cell nucleus. These phenomena are probably necessary for the exhibition of MP-4 differentiating activity.

On the possible manifestation of harmonic-anharmonic dynamical transition in glassy media in electron paramagnetic resonance of nitroxide spin probes.

Continuous wave (cw) electron paramagnetic resonance (EPR) and echo-detected (ED) EPR were applied to study molecular motions of nitroxide spin probes in glassy glycerol and o-terphenyl. A linear decrease with increasing temperature of the total splitting in the cw EPR line shape was observed at low temperatures in both solvents. Above some temperature points the temperature dependencies become sharper. Within the model of molecular librations, this behavior is in qualitative and quantitative agreement with the numerical data on neutron scattering and Mossbauer absorption for molecular glasses and biomolecules, where temperature dependence of the mean-squared amplitude of the vibrational motion was obtained. In analogy with these data the departure from linear temperature dependence in cw EPR may be ascribed to the transition from harmonic to anharmonic motion (this transition is called dynamical transition). ED EPR spectra were found to change drastically above 195 K in glycerol and above 245 K in o-terphenyl, indicating the appearance of anisotropic transverse spin relaxation. This appearance may also be attributed to the dynamical transition as an estimation shows the anisotropic relaxation rates for harmonic and anharmonic librational motions and because these temperature points correspond well to those known from neutron scattering for these solvents. The low sensitivity of ED EPR to harmonic motion and its high sensitivity to the anharmonic one suggests that ED EPR may serve as a sensitive tool to detect dynamical transition in glasses and biomolecules.

Restricted orientational motion of nitroxides in molecular glasses: direct estimation of the motional time scale basing on the comparative study of primary and stimulated electron spin echo decays.

A comparative study of anisotropic relaxation in two-pulse primary and three-pulse stimulated electron spin echo decays provides a direct way to distinguish fast (correlation time tau(c)<10(-6) s) and slow (tau(c)>10(-6) s) motions. Anisotropic relaxation is detected as a difference of the decay rates for different resonance field positions in anisotropic electron paramagnetic resonance spectra. For fast motion anisotropic relaxation influences the primary echo decay and does not influence the stimulated echo decay. For slow motion it is seen in both two-pulse echo and three-pulse stimulated echo decays. For nitroxide spin probes dissolved in glassy glycerol only fast motion was found below 200 K. Increase of temperature above 200 K results in the appearance of slow motion. Its amplitude increases rapidly with temperature increase. While in glycerol glass slow motion appears above glass transition temperature T(g), in ethanol glass it is observable below T(g). The scenario of motional dynamics in glasses is proposed which involves the broadening of the correlation time distribution with increasing temperature.

Induction of differentiation in leukemic cell strains with myelopeptide-4.

We studied the capacity of myelopeptide-4 (regulatory peptide of the bone marrow origin) to induce terminal differentiation of HL-60 and K-562 leukemic cells. Myelopeptide-4 increased the expression of CD14 and CD38 differentiation antigens on the surface of HL-60 cells and of CD44 antigen on K-562 cells, induced the appearance of mature monocyte/macrophages in HL-60 culture and hemoglobin-producing cells in K-562 cell culture, and stimulated phagocytic activity of THP-1 leukemic cells. Myelopeptide-4 is an endogenous factor of cell differentiation, a prospective agent for antileukemic therapy.

Effect of myelopeptide-2 on the development of spontaneous and urethane-induced tumors in mice.

We studied the effect of endogenous immunoregulatory myelopeptide-2 on the development of spontaneous and urethane-induced lung adenomas. Long-term treatment with myelopeptide-2 (25 injections) in parallel with urethane poisoning decreased animal mortality caused by this carcinogen. Short-term treatment with myelopeptide-2 decreased the number of spontaneous and urethane-induced lung tumors in mice. Myelopeptide-2 delayed the appearance of urethane-induced tumors irrespective of the number of injections.

Immunoregulatory properties of hexapeptide isolated from porcine bone marrow cell culture.

Myelopeptide 1 (MP-1) is hexapeptide originally isolated from porcine bone marrow cell culture. It was synthesized and its immunoregulatory properties were studied. MP-1 caused a 1.5-2-fold dose-dependent increase of antibody production in the culture of mouse immune lymph node cells. It abolished Con A induction of T suppressors in the suspension of mouse spleen cells and counteracted the inhibitory effect of T suppressors on antibody production. The inoculation of MP-1 (1 x 10(-9) g/mouse) to mice two weeks after their gamma-irradiation (2 Gy) resulted in an increase of antibody production up to 80.2 +/- 15.5% as compared to that in the irradiated control 37.6 +/- 12.0%. Immunofluorescent analysis revealed the specific binding of MP-1 with receptors on the target cells in the suspension of mouse spleen cells. It is supposed that MP-1 participates in the immunoregulatory processes in the living organism.

Myelopeptide-1 blocks the T-suppressor activity.

The immunoregulatory properties of two myelopeptides (MPs) [Phe-Leu-Gly-Phe-Pro-Thr (MP-1) and Leu-Val-Val-Tyr-Pro-Trp (MP-2)] were studied. The antibody production-stimulating activity was determined in the culture of lymph node cells (LNC) obtained from mice on the 4th day of the secondary immune response to sheep red blood cells (SRBC). T suppressors were induced in mouse spleen cell (SC) culture by incubation with concanavalin A (Con A) for 48 h. Immune LNC were cultured alone or with T suppressors in the presence or absence of MPs. After cultivation, the immune response was estimated by enumeration of indirect antibody forming cells (AFC). MP-1, but not MP-2, increased the number of AFC and abolished Con A induction of T suppressors in SC suspensions. MP-1 also fully blocked and MP-2 only partly decreased the inhibitory effect of T suppressors in the culture of immune LNC. Immunoregulatory peptide MP-1 may participate in regulation of the antibody response and in development of some states of immunological tolerance.

[Prevention and correction of stress-induced immunodeficiency by atrial electroacupuncture]. / Profilaktika i korrektsiia vyzvannogo stressom immunodefitsitnogo sostoianiia metodom aurikuliarnoi élektroakupunktury.

Secondary immunodeficiency characterized by a sharp decrease in the number of antibody-forming cells in the lymph organs developed in mice under conditions of an immunosuppressive effect of swim stress and hypobaric hypoxia. Auricular electroacupuncture (AEAP) preceding swim stress blocks its immunosuppressive effect completely. AEAP possesses a high preventive action and also produces a marked corrective effect in development of stress-induced immunodeficiency. For instance, AEAP applied in the productive phase of the immune response restores effectively immunocompetence of the body subjected to hypobaric stress. In applying AEAP, the coefficients of antibody-production stimulation in the cells of the lymph nodes, calculated in relation to the hypoxic control, are 3.5-4.0, which do not yield to the values recorded with the use of the most effective pharmacological immunomodulators. AEAP may be considered a promising measure for the prevention and correction of immunodeficiency caused by stress.