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Goufmana E.I.,Scientific Research Institute of Physical Chemical Medicine | Tikhonov N.B.,Russian Academy of Medical Sciences | Lokshind A.E.,University of Pittsburgh
Cancer Biomarkers | Year: 2015

BACKGROUND: The investigation of autoantibodies which may play a role in the processes of angiogenesis and tumorogenesis is important in the early diagnostis of cancer. OBJECTIVE: This study aimed to investigate the levels of autoantibodies to Glu-plasminogen (Pg) in plasma of patients with tumors. METHODS: Plasma samples from healthy volunteers were compared with samples from patients with prostate cancer using 2D electrophoresis and MALDI-TOF mass spectrometry. Plasma samples from 25 patients with prostate cancer, 15 patients with benign prostatic hyperplasia (BPH), 29 patients with breast cancer, and 43 healthy volunteers were tested using ELISA to anti-Pg IgG autoantibodies. Affinity chromatography on Pg-sepharoses was used to assess the quantity of anti-Pg IgG in control plasma and plasma of prostate cancer patients. ATTESTAT program was used for nonparametric analysis. RESULTS: Using 2D electrophoresis, marker spots below 50 kD were detected in prostate cancer samples. These spots were identified as fragments of Pg and IgG. Using affinity chromatography on Pg-sepharose, the quantity of IgG bound to Pg versus total IgG was determined to be 9% in control and 27% in prostate cancer samples. The frequency of occurence of elevated levels of anti-Pg IgG was 84% in prostate cancer samples, 69% in breast cancer samples, 40% in BPH samples, and 11% in healthy plasma. CONCLUSIONS: Autoantibodies to Pg may be involved in tumorogenesis and elevated levels of anti-Pg IgG antibodies may be a risk factor for tumor development. © 2015 - IOS Press and the authors. All rights reserved.

Petrova E.K.,Moscow State University | Nikitin N.A.,Moscow State University | Trifonova E.A.,Moscow State University | Protopopova A.D.,Scientific Research Institute of Physical Chemical Medicine | And 2 more authors.
Biochimie | Year: 2015

Filamentous helical Potato virus X (PVX) can be regarded as one of the well-studied viruses. Nevertheless, some aspects of the PVX assembly remained obscure. Previously, we have shown that the presence of a cap structure at the 5′ end of PVX RNA is indispensable for assembly of viral ribonucleoprotein (vRNP) particles varying in length. Here, most significantly, removal of the cap structure from previously capped PVX RNA did not affect the efficiency of decapped RNA molecules to be assembled into vRNP. This result provided evidence that the cap structure by itself does not act as a signal for initiation of vRNP assembly. These observations allowed to presume that the capping triggers some spatial changes in the 5'-proximal site of PVX RNA creating a "conformational encapsidation signal for vRNP assembly", which is capable of triggering vRNP assembly in the absence of cap structure. Apparently, during capping the 5'-proximal segment of PVX RNA acquires a unique conformation which is stable to be retained even after cap removal. © 2015 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

Dobretsov G.E.,Scientific Research Institute of Physical Chemical Medicine
Biophysics (Russian Federation) | Year: 2013

A brief history of the development of the method of fluorescent probes and examples of its application are presented. Works done at the 2nd Moscow medical institute and institute of physical chemical medicine in collaboration with other institutes on: (1) detection of T- and B-lymphocytes in immune pathology; (2) investigation of the structure and clinical estimation of lipoproteins from blood plasma or serum in relation to the assessment of risk factors for the development of cardiovascular diseases; (3) detection of changes in album molecule in a series of pathological processes improving the prognosis of the development of such diseases as peritonitis, pancreatitis, poisoning with psychotropic preparations etc.; (4) intravital measurement of the potentials in electric fields in leukocytes and changes of these fields in the course of immunological diseases are described. With these approaches it is possible to study molecular events in the course of pathogenesis and also obtain diagnostically significant information on physical chemical aspect of these events. This information is not a conventional method used in the clinical laboratory. © 2013 Pleiades Publishing, Inc.

Balabushevich N.G.,Moscow State University | Pechenkin M.A.,Moscow State University | Shibanova E.D.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Volodkin D.V.,Fraunhofer Institute for Biomedical Engineering | Mikhalchik E.V.,Scientific Research Institute of Physical Chemical Medicine
Macromolecular Bioscience | Year: 2013

Multicomponent insulin-containing microparticles are prepared by layer-by-layer assembly of dextran sulfate and chitosan on the core of protein-polyanion complex with or without protease inhibitors. Oral bioavailability of the encapsulated insulin is improved due to the cumulative effect of each component. A physico-chemical study shows that the particle design allows adjustment of the pH-dependent profile of the insulin release, as well as mucoadhesive properties and Ca2+ binding ability of the microparticles. Supplementing the microparticles with 2-3% protease inhibitors fully prevents proteolysis of human insulin. The pharmacological effect of microencapsulated insulin in doses 50-100 IU kg-1 is demonstrated in chronic experiments after oral administration to diabetic rats fed ad libitum. Insulin is encapsulated in multicomponent polymer microparticles by layer-by-layer assembly. Systematic studies of particle physicochemical properties at certain biological conditions demonstrate an improvement of oral insulin bioavailability. The pharmacological effect of microencapsulated insulin for per os in doses 50-100 IU kg-1 is shown in chronic experiments on diabetic rats fed ad libitum. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Protopopova A.D.,Scientific Research Institute of Physical Chemical Medicine | Barinov N.A.,Scientific Research Institute of Physical Chemical Medicine | Zavyalova E.G.,Moscow State University | Kopylov A.M.,Moscow State University | And 2 more authors.
Journal of Thrombosis and Haemostasis | Year: 2015

Background: Fibrinogen has been intensively studied with transmission electron microscopy and x-ray diffraction. But until now, a complete 3D structure of the molecule has not yet been available because the two highly flexible αC regions could not be resolved in fibrinogen crystals. This study was aimed at determining whether the αC regions can be visualized by high-resolution atomic force microscopy. Methods: Atomic force microscopy with super high resolution was used to image single molecules of fibrinogen and fibrin associates. The key approach was to use a graphite surface modified with the monolayer of amphiphilic carbohydrate-glycine molecules and unique supersharp cantilevers with 1 nm tip diameter. Results: Fibrinogen αC regions were visualized along with the complete domain structure of the protein. In almost all molecules at pH 7.4 the D domain regions had one or two protrusions of average height 0.4 ± 0.1 nm and length 21 ± 6 nm. The complex, formed between thrombin and fibrinogen, was also visualized. Images of growing fibrin fibers with clearly visible αC regions have been obtained. Conclusions: Fibrin αC regions were visible in protofibrils and large fibers; αC regions intertwined near a branchpoint and looked like a zipper. These results support the idea that αC regions are involved in the thickening of fibrin fibers. In addition, new details were revealed about the behavior of individual fibrin molecules during formation of the fibrin network. Under the diluted condition, the positioning of the αC regions could suggest their involvement in long-range interactions between fibrin but not fibrinogen molecules. © 2014 International Society on Thrombosis and Haemostasis.

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