A role of methionines in the functioning of oxidatively modified fibrinogen.

Posttranslational modifications in fibrinogen resulting from induced oxidation or oxidative stress in the organism can have deleterious influence on optimal functioning of fibrinogen, causing a disturbance in assembly and properties of fibrin. The protective mechanism supporting the ability of fibrinogen to function in ROS-generating environment remains completely unexplored. The effects of very low and moderately low HOCl/-OCl concentrations on fibrinogen oxidative modifications, the fibrin network structure as well as the kinetics of both fibrinogen-to-fibrin conversion and fibrin hydrolysis have been explored in the current study. As opposed to 25 Μm, HOCl/-OCl, 10 µM HOCl/-OCl did not affect the functional activity of fibrinogen. It is shown for the first time that a number of Met residues, AαMet476, AαMet517, AαMet584, BßMet367, γMet264, and γMet94, identified in 10 µM HOCl/-OCl fibrinogen by the HPLC-MS/MS method, operate as ROS scavengers, performing an important antioxidant function. In turn, this indicates that the fibrinogen structure is adapted to the detrimental action of ROS. The results obtained in our study provide evidence for a protective mechanism responsible for maintaining the structure and functioning of fibrinogen molecules in the bloodstream under conditions of mild and moderate oxidative stress.

Peroxide-Induced Damage to Plasminogen Molecules.

Plasminogen is a zymogenic form of plasmin, an enzyme that plays a fundamental role in the dissolution of fibrin clots as well as in many other physiological processes. For the first time, by the method of gas chromatography-mass spectrometry, post-translational modifications in the primary structure of plasminogen treated with physiologically relevant amounts of hydrogen peroxide were identified. It was found that methionine and tryptophan residues located in different structural regions of plasminogen served as targets of the oxidant. Plasminogen oxidation caused a dose-dependent effect in decreasing the fibrinogenolytic activity of plasmin evidenced by the formation of fibrinogen degradation products. The possible antioxidant role of methionines in the oxidative modification of plasminogen is discussed.

The Nature of Resistance of the Coagulation Factor XIII Structure to Hypochlorite-Induced Oxidation.

The damage to blood coagulation factor XIII (FXIII) at different stages of its enzymatic activation under the action of various physiological amounts of hypochlorite ion was studied. The results obtained by HPLC-MS/MS, SDS-PAGE, and colorimetry showed that, during the conversion of FXIII to FXIIIa, the vulnerability of FXIII to hypochlorite-induced oxidation increased. FXIII oxidized with 150 µM hypochlorite completely retained its enzymatic activity inherent to the intact protein, whereas FXIIIa treated with 50 µM hypochlorite showed sharply reduced enzymatic activity. It was shown that a number of methionine and cysteine residues on the catalytic subunit can perform antioxidant function; additionally, the regulatory subunits of FXIII-B contribute to the antioxidant protection of the catalytic center of the FXIII-A subunit, which, together with the tight packing of the tetrameric structure of the FXIII proenzyme, are the three factors that provide high protein resistance to the oxidizing agent.

[Forum of military physicians of Asia Pacific region].

Forum of military physicians of Asia Pacific region. Authors proviae a report about the 3rd Military medical Asia Pacific congress, which was held in August 2016 in Saint Petersburg under the authority of the International military medical committee. Within the frame of the congress was held regional workshop, in the course of which was heard a report of the Thailand representative, requests of observer states for the entry. The country, which will hold the 4th Military medical Asia Pacific congress, is Iran. On the last day of the congress delegates visited the finals of the Military-medical race of the military physicians profession skill competition within <.

[Legal regulation of clinical drug trials for the use in military-medical organisations].

The authors identified problematic issues of legal regulation of clinical drug trials for medical use, and proposed possible solutions. It has been established that the conduction of clinical trials, of medicinal products is based on the norms of various branches of law embodied in the Constitution of the Russian Federation, the norms of international law, the Civil Code of the Russian Federation and federal laws and subordinate legislations regulating health and pharmaceutical activity. According to the authors, the norms of bioethics can be attributed to the sources of legal doctrine. It is proposed to oblige executives of clinical trials to make a report about effectiveness and safety of drugs and pass the results to the customer, in his/her turn the customer is obliged to accept the results of these trials and pay for them.

The radioisotope complex project "RIC-80" at the Petersburg Nuclear Physics Institute.

The high current cyclotron C-80 capable of producing 40-80 MeV proton beams with a current of up to 200 µA has been constructed at Petersburg Nuclear Physics Institute. One of the main goals of the C-80 is the production of a wide spectrum of medical radionuclides for diagnostics and therapy. The project development of the radioisotope complex RIC-80 (radioisotopes at the cyclotron C-80) at the beam of C-80 has been completed. The RIC-80 complex is briefly discussed in this paper. The combination of the mass-separator with the target-ion source device, available at one of the new target stations for on-line or semi on-line production of a high purity separated radioisotopes, is explored in greater detail. The results of target and ion source tests for a mass-separator method for the production of high purity radioisotopes (82)Sr and (223,224)Ra are also presented.

Binding energies of the ground triplet state a(3) Σ(u)(+) of Rb2 and Cs2 in terms of the generalized Le Roy-Bernstein near-dissociation expansion.

Formulae of Le Roy-Bernstein near-dissociation theory are derived in a general isotope-invariant form, applicable to any term in the rotational expansion of a diatomic ro-vibrational term value. It is proposed to use the generalized Le Roy-Bernstein expansion to describe the binding energies (ro-vibrational term values) of the ground triplet state a(3) Σ(u)(+) of alkali metal dimers. The parameters of this description are determined for Rb2 and Cs2 molecules. This approach gives a recipe to calculate the whole variety of the binding energies with characteristic accuracies from ∼1 × 10(-3) to 1 × 10(-2) cm(-1) using a relatively simple algebraic equation.

Updated potential energy function of the Rb2 a(3)Σu(+) state in the attractive and repulsive regions determined from its joint analysis with the 2(3)Π0g state.

We report new experimental data for the Rb2 a(3)Σu(+) and 2(3)Π0g states obtained using the Perturbation Facilitated Infrared-Infrared Double Resonance (PFIIDR) technique. The results include ro-vibrational term values of the 2(3)Π0g state and resolved fluorescence spectra of the 2(3)Π0g→a(3)Σu(+) transitions for a wide range of rotational and vibrational quantum numbers. An analysis of these data confirms the initial assignment of the transitions to the a(3)Σu(+) state reported in our earlier work [B. Beser, V. B. Sovkov, J. Bai, E. H. Ahmed, C. C. Tsai, F. Xie, L. Li, V. S. Ivanov, and A. M. Lyyra, J. Chem. Phys. 131, 094505 (2009)]. The potential energy functions of the Rb2 a(3)Σu(+) and 2(3)Π0g states are derived from a simultaneous fit of the available experimental data. The improved potential function of the Rb2 a(3)Σu(+) state spans both the attractive and repulsive regions starting with internuclear distance R ∼ 4.5 Å.

New laser setup for the selective isotope production and investigation in a laser ion source at the IRIS (Investigation of Radioactive Isotopes on Synchrocyclotron) facility.

New laser installation for the resonance ionization spectroscopy in a laser ion source and for rare isotope production has been recently put into operation at the IRIS (Investigation of Radioactive Isotopes on Synchrocyclotron) facility (Petersburg Nuclear Physics Institute, Gatchina). This is a significant improvement of a previous target-laser ion source device of the IRIS mass-separator, working on-line with 1 GeV proton beam of PNPI (Petersburg Nuclear Physics Institute) synchrocyclotron. It makes possible for us to get the isobarically clean radioactive isotope beams of a great number of chemical elements. New laser setup provides the two- or three-resonance step ionization in the range of wavelength of 265-850 nm. The first results obtained at the laser setup for Tl isotopes are presented.

Joint analysis of the Cs2 a3Σu+ and 1g (3(3)Π1g) states.

Sets of experimental data on the Cs(2) a(3)Σ(u)(+) and 1(g) (3(3)Π(1g)) states, including the bound-bound and bound-free fluorescence spectra, are analyzed simultaneously to produce the potential energy curves of both states in the form of the Morse long range multiparameter function. The attractive branch of the a(3)Σ(u)(+) state potential is improved relative to the one reported in our earlier work [F. Xie, V. B. Sovkov, A. M. Lyyra, D. Li, S. Ingram, J. Bai, V. S. Ivanov, S. Magnier, and L. Li, J. Chem. Phys. 130, 051102 (2009)], in which the data on this state alone were analyzed. Besides, the new potential of this state also includes the repulsive branch in the range spanned by the bound-free fluorescence spectra. We have not found experimental evidence of the double minimum character of the 3(3)Π(1g) state potential, predicted by ab initio calculations, at least up to v = 8. This fact testifies that the upper state observed is better described by the Hund coupling case (c), in which the case (a) electronic basis states are intermixed by the strong spin-orbit interaction.

[Plasma lipoproteins as drug carriers. Effect of phospholipid formulations].

The extensive development of nanotechnologies in the last two decades has brought about new understanding of plasma lipoproteins (LP) as natural drug nanocarriers that escape interaction with immune and reticuloendothelial systems. Drugs bound to LP (especially LDL) can more actively penetrate into cells of many cancer and inflammation tissues with enhanced expression or/and dysregulation of B,E receptors or possibly scavenger SR-BI receptors. Relevant studies are focused on the development of new dosage forms by conjugating lipophilic drugs either with isolated plasma LP or with their model formulations, such as nanoemulsions, mimetics, lipid nanospheres, etc. Some authors include in these particles serum or recombinant apoproteins, peptides, and modified polymer products. As shown recently, protein-free lipid nanoemulsions in plasma take up free apoA and apoE. Complexes with various LP also form after direct administration of lypophilic drugs into blood especially those enclosed in phospholipid formulations, e.g. liposomes. Results of evaluation of some lipophilic dugs (mainly cytostatics, amphotericin B, cyclosporine A, etc.) are discussed. Original data are presented on the influence of phospholipid formulations on the distribution of doxorubicin and indomethacin between LP classes after in vitro incubation in plasma. On the whole, the review illustrates the importance of research on LP and phospholi pid forms as drug nanocarriers to be used to enhance effect of therapy.

Experimental investigation of the 85Rb2 a 3Sigma(u)+ triplet ground state: multiparameter Morse long range potential analysis.

Using perturbation facilitated infrared-infrared double resonance excitation of the (85)Rb(2) molecule, we have observed spectrally resolved fluorescence to the a (3)Sigma(u)(+) state. We have analyzed the rovibrational energy level structure of the (85)Rb(2) a (3)Sigma(u)(+) state and derived a multiparameter Morse Long Range (MLR) potential and molecular constants for this state, which can be used to predict term values without needing to solve the radial Schrödinger equation.

Experimental investigation of the Cs2 a 3Sigma(u)+ triplet ground state: multiparameter Morse long range potential analysis and molecular constants.

We have observed the vibrational levels v(") = 0-40 of the Cs(2) a (3)Sigma(u)(+) state by perturbation facilitated infrared-infrared double resonance excitation and spectrally resolved fluorescence measurements, and derived a multiparameter Morse long range potential and molecular constants based on these data.

[Effects of (22S,23S)-3beta-hydroxy-22,23-oxido-5alpha-ergost-8(14)-en-15-one and (22R,23R)-3beta-hydroxy-22,23-oxido-5alpha-ergost-8(14)-en-15-one on lipid biosynthesis and exogeneous cholesterol metabolism in hep G2 cells].

Novel synthetic oxysterols (22S,23S)-3beta-hydroxy-22,23-oxido-5alpha-ergost-8(14)-en-15-one (I) and (22R,23R)-33-hydroxy-22,23-oxido-5alpha-ergost-8(14)-en-15-one (II) efficiently inhibited cholesterol biosynthesis in human hepatoma Hep G2 cell line at a short time incubation in a serum free medium (IC50 = 1.9 +/- 0.2 and 0.6 +/- 0.2 microM, respectively). Cultivation of Hep G2 cells in the presence of compound 5 microM concentration of both (1) and (II), led to significant depression of cholesterol biosynthesis (52% and 57% from control), and remarkable changes in fatty acids, triglycerides, and cholesteryl esters biosynthesis. Compounds (I) and (II) stimulated transformation of exogeneous cholesterol to polar products secreted into the culture medium (156% and 175% from control), that was shown in experiments in Hep G2 cells prelabeled with [3H]cholesterol.

The K2 2(3)Pi(g) state: new observations and analysis.

Totally 3045 transitions into the 2(3)Pi(g) v = 0-42, J = 0-103, Omega = 0, 1, 2 rovibrational levels have been observed by infrared-infrared double resonance fluorescence excitation and two-photon spectroscopy. Molecular constants including the spin-orbit interaction parameters are obtained. Although the K2 2(3)Pi(g) state dissociates to the 4s + 3d atomic limit, it is strongly mixed with the 3P ionic states in the range of the potential well. This mixing results in a relatively large equilibrium internuclear distance Re = 5.254 A and a larger spin-orbit constant A0 approximately 14.17 cm(-1) than that of the atomic limit -2.33 cm(-1). Strong perturbations of the 2(3)Pi(g) levels observed are attributed to the spin-orbit coupling with the 4(1)Sigma(g)+ state.

Experimental study of the 39K2 2 3Pi(g) state by perturbation facilitated infrared-infrared double resonance and two-photon excitation spectroscopy.

The 39K2 2 3Pi(g) state has been observed by perturbation facilitated infrared-infrared double resonance and two-photon excitations. The vibrational numbering of the 2 3Pi(g) levels was determined by resolved fluorescence into the bound levels as well as to the continuum of the a 3Sigma(u)+ state. The rotational assignment of the 2 3Pi(g) levels excited by two-photon transitions was determined from excitation frequencies and resolved fluorescence into the bound levels of the a 3Sigma(u) + and b 3Pi(u) states. Molecular constants obtained from these observed levels agree with theoretical constants.

Tumor cell cytolysis mediated by valorphin, an opioid-like fragment of hemoglobin beta-chain.

Valorphin, an endogenous opioid-like hemoglobin fragment, is cytotoxic for L929 and K562 tumor cells in 10(-7)-10(-13) M concentration range. Because cytolytic effects induced by valorphin in K562 cells are inhibited by naloxone, opioid receptors should be involved in induction of valorphin-mediated tumor cell death. Three distinct cytolytic processes, differing in the onset time and the development time, take place with K562 cells within 10-18 h of incubation with valorphin. All three processes are not associated with apoptotic mechanism of cell death.

[Identification of immunoreactive epitopes in proteins coded by gag, env, and pol genes of the type I human T-lymphotropic virus (HTLV-I) using synthetic peptides]. / Identifikatsiia immunoreaktivnykh épitopov v belkakah, kodiruemykh genami gag, env i pol T-limfotropnogo virusa cheloveka tipa I (HTLV-I) s ispol'zovaniem sinteticheskikh peptidov.

Reactivity of 26 synthetic peptides that comprise 12 to 26 amino acid residues corresponding to segments of the gag p19, env gp46, and pol proteins of human T-lymphotropic virus type I toward 31 positive sera was studied using enzyme-linked immunosorbent assay. Specific reactivity with high titers of antibodies (presented in reciprocal dilution values) was detected for the synthetic peptides corresponding to fragments 110-130 and 100-130 (titers up to 4050) of p19, 174-197 (up to 800), 186-201 (up to 4050), 191-215 (up to 1350), 242-257 (up to 800), and 272-292 (up to 450) of gp46. Immunoreactivity of seven peptides, fragments of pol-proteins, was weak. New linear epitopes in the regions 145-158, 272-277, and 292-300 of gp46 were detected. In addition, location of the known linear epitopes in p19 and gp46 was refined on the basis of comparative study of overlapping peptides from these proteins.

Peptides of a major histocompatibility complex class I (Kb) molecule cause prolongation of skin graft survival and induce specific down-regulatory T cells demonstrable in the mixed lymphocyte reaction.

Six individual peptides of the major histocompatibility complex (MHC) class I molecule H-2Kb were synthesized. Intravenous injection of peptide 6 into mice prolonged the survival of Kb (BL/6 or B10.MBR) skin grafts on allogeneic R101 and B10.AKM mice, respectively. This was specific, as control skin grafts from Kk (B10.BR) or Kd (DBA/2) donors, respectively, were rejected at the same time in both control and peptide-treated mice. The optimal doses for peptide 6, which is from the alpha 2 domain, were defined. The test system was the inhibition of proliferation in vitro of naive lymph node cells by syngeneic mitomycin c-treated spleen cells from R101 mice preimmunized with irradiated stimulator splenocytes of Kb (BL/6) origin. Down-regulation was specific, as proliferation in response to third-party allogeneic stimulator Kk (B10.BR) splenocytes was not inhibited. Of the six peptides of H-2Kb tested, potent down-regulatory cells were induced by peptides 2 (alpha 1 domain) and 5 and 6 (alpha 2 domain). The greatest down-regulatory activity was obtained by giving peptide 2 to mice that had already been immunized against H-2Kb by injecting EL4 cells. Under the same conditions, injecting peptide 2 did not induce any cytotoxic T cells. In contrast, specific cytotoxic lymphocytes (CTL) were induced when cells from primed mice were incubated for 4 days with heated stimulator cells from BL/6 mice. The data suggest that peptides from MHC class I molecules activate precursors of down-regulatory T cells, but not of CTL, and this may explain their ability to prolong skin allograft survival.