Biochemistry (Moscow), Supplement Series B: Biomedical Chemistry (v.8, #1)
Mass spectrometry analysis of blood low-molecular fraction as a method for unification of therapeutic drug monitoring by P. G. Lokhov; D. L. Maslov; O. P. Trifonova; E. E. Balashova; A. I. Archakov (1-10).
The review describes a new method of therapeutic drug monitoring (TDM) based on direct infusion of low-molecular fraction of blood into an electrospray ionization source of mass spectrometer. This technique allows performing TDM of almost all drugs used in clinical practice. Universality and a high-throughput mode of the method significantly simplify wide application of this method. Moreover, the possibility of method application in most cases of drug therapy has been argued as a tool for control of drug doses, rationality of drug therapy, and quality of drugs used. In conclusion, prospects for application of the method as primary means of improving the quality and personalization of drug therapy have been discussed.
Keywords: mass spectrometry; therapeutic drug monitoring (TDM); capillary blood; personalized drug therapy; direct mass spectrometry
Defensins are natural peptide antibiotics of higher eukaryotes by D. V. Grishin; N. N. Sokolov (11-18).
The goal of this review is to characterize defensins representing an evolutionary the most ancient family of antimicrobial peptides. It gives general information on functional and structural features of defensins as the main components of the first-line defense of higher eukaryote organisms against infectious agents. The review not only considers current situation in the defensin research but also perspectives of creation of recombinant antimicrobial peptides of biomedical application.
Keywords: antimicrobial peptides; defensins; mechanism of bactericidal action of defensins; recombinant antimicrobial peptides
Prospects for the use of peptides and their derivatives, structurally corresponding to the g protein-coupled receptors, in medicine by A. O. Shpakov; E. A. Shpakova (19-26).
The regulation of signaling pathways involved in the control of many physiological functions is carried out via the heterotrimeric G protein-coupled receptors (GPCR). The search of effective and selective regulators of GPCR and intracellular signaling cascades coupled with them is one of the important problems of modern fundamental and clinical medicine. Recent data suggest that synthetic peptides and their derivatives, structurally corresponding to the intracellular and transmembrane regions of GPCR, can interact with high efficiency and selectivity with homologous receptors and influence the functional activity of intracellular signaling cascades and fundamental cellular processes controlled by them. GPCR-derived peptides are active in both in vitro and in vivo. They regulate hematopoiesis, angiogenesis and cell proliferation, inhibit tumor growth and metastasis, and prevent the inflammatory diseases and septic shock. These data show great prospects in the development of the new generations of drugs based on GPCR-derived peptides, which can regulate the important functions of the organism.
Keywords: angiogenesis; metastasis; tumor; pepducin; peptide; sepsis; G protein-coupled receptor; transmembrane region; thrombosis; intracellular loop
Potentiation of YC-1 activation of soluble guanylate cyclase by NO donors and the increase of the synergistic effect of YC-1 on the NO-dependent activation of the enzyme by 1,2,3-triazolyl-1,2,5-oxadiazole derivatives by I. S. Severina; N. V. Pyatakova; A. Yu. Shchegolev; V. Yu. Rozhkov; L. V. Batog; N. N. Makhova (27-33).
The influence of (1H-1,2,3-triazol-1-yl)-1,2,5-oxadiazole derivatives: 4-amino-3-(5-methyl-4-ethoxycarbonyl-(1H-1,2,3-triazol-1-yl)-1,2,5-oxadiazole (TF4CH3) and 4,4′-bis(5-methyl-4-ethoxycarbo-nyl-1H-1,2,3-triazol-1-yl)-3,3′-azo-1,2,5-oxadiazole (2TF4CH3) on stimulation of human platelet soluble guanylate cyclase by YC-1, NO donors (sodium nitroprusside, SNP, and spermine NONO) and on a synergistic increase of NO-dependent activation of the enzyme in the presence of YC-1 has been investigated. Both compounds increased guanylate cyclase activation by YC-1, potentiated guanylate cyclase stimulation by NO donors and increased the synergistic effect of YC-1 on the NO-dependent activation of soluble guanylate cyclase. The similarity in the properties of the examined 1,2,3-triazol-1-yl-1,2,5-oxadiazole derivatives with that of YC-1 and a possible mechanism underlying the recognized properties of compounds used are discussed.
Keywords: soluble guanylate cyclase; nitric oxide; YC-1; (1H-1,2,3-triazol-1-yl)-1,2,5-oxadiazole derivatives
Differences of antioxidant systems in the cerebellum and hippocampus by E. A. Kosenko; E. E. Beloushko; Y. G. Kaminsky (34-36).
The effect of portacaval shunting on the antioxidant status of the cerebellum and hippocampus has been studied in rats using standard methods of enzymatic analysis. Endogenous ammonia levels and activities of eight antioxidant enzymes were shown to be unequal in these two brain regions and to respond differently to intrahepatic portosystemic shunt surgery.
Keywords: portacaval shunt; antioxidant enzymes; hippocampus; cerebellum
The study of the neuroprotective activity of the apolipoprotein E peptide mimetic Cog1410 in transgenic strains of Drosophila melanogaster by E. M. Latypova; S. I. Timoshenko; G. A. Kislik; M. P. Vitek; A. L. Schwarzman; S. V. Sarantseva (37-42).
The neuroprotective activity of peptide mimetic of apolipoprotein E (apoE) Cog1410, containing the amino acid sequence of the apoE receptor-binding domain, has been investigated in transgenic Drosophila melanogaster strains expressing human APP and beta-secretase genes. Expression of double transgenes caused neuropathological processes typical for Alzheimer’s disease (AD): neurodegeneration, cognitive impairments, and amyloid deposition in the brain. It was shown that Cog1410 reduces neurodegeneration in the brain of transgenic flies and improves cognitive functions (odor recognition). These data suggest that Cog1410 is a potential neuroprotector that can be used in AD treatment.
Keywords: ApoE; Cog1410; Alzheimer disease; Drosophila melanogaster
The effect of docosahexaenoic acid moiety on the cytotoxic activity of 1,2,4-thiadiazole derivatives by M. G. Akimov; N. M. Gretskaya; V. A. Karnoukhova; I. V. Serkov; A. N. Proshin; V. Yu. Shtratnikova; V. V. Bezuglov (43-46).
Among 3-(2-aminopropyl)-1,2,4-thiadiazole derivatives containing substitutable secondary amino group and exhibiting cytotoxic activity towards rat C6 glioma cells three compounds with LD50 values ranged from 6 to 48 μM have been chosen. For these compounds amides with docosahexaenoic acid were synthesized and their cytotoxic activity was studied. It was shown that, although docosahexaenoic acid itself was not toxic for C6 glioma cells, its addition to the amino derivatives of 1,2,4-thiadiazole increased or decreased their cytotoxicity. These results demonstrate that acylation of cytotoxic compounds with docosahexaenoic acid does not necessarily lead to the increase of their activity and sometimes can inactivate a parent compound. This fact should be taken into consideration because of possibility of biological attachment of docosahexaenoic acid to the amino group of anti-cancer drugs.
Keywords: docosahexaenoic acid; 1,2,4-thiadiazoles; rat C6 glioma
Monoclonal antibodies to tumor necrosis factor-alpha influence metabolic activity in lymphocytes under conditions of chronic ethanol consumption by M. V. Haretskaya; V. M. Sheibak (47-52).
In Wistar rats consuming ethanol for 10 weeks (the Lieber-De Carli liquid diet), dose-dependent effects of infliximab (monoclonal antibody to tumor necrosis factor alpha, TNFα) have been demonstrated. The 10-day course of intraperitoneal injections of infliximab (1 or 10 mg/kg body weight) to rats exposed to chronic ethanol consumption caused changes in the concentrations of free amino acids and their nitrogencontaining metabolites, and also promoted the development of amino acid imbalance in liver lymphocytes. Treatment of ethanol consuming animals with infliximab increased in a dose-dependent manner the pool of free amino acids and the amount of their nitrogen-containing metabolites in lymphocytes. Adaptive changes in the system of the transport of amino acids and other nutrients in the liver lymphocytes are one of the possible mechanisms associated with TNFα inactivation.
Keywords: rat; amino acids; lymphocytes; ethanol; antibodies to TNFα
Prediction of the action of ligands of steroid hormone receptors by I. V. Fedyushkina; I. V. Romero Reyes; A. A. Lagunin; V. S. Skvortsov (53-58).
Several predictive models have been created using two main calculation methods, PASS and SIMCA. These models predict a type of a physiological response of steroid compounds induced by their binding to nuclear receptors of steroid hormones. Ten models of agonists and antagonists of estrogen, progesterone, androgen, glucocorticoid and mineralocorticoid receptors have been considered. Two variants have been used in the case of SIMCA (the Dragon descriptors and indices of similarity); these included molecular and constitutive features and indices of similarity, respectively. It has been shown that using discriminant analysis and a structural formula of the compound of interest it is possible to predict a spectrum of its actions with an average accuracy of 80–85%.
Keywords: steroid hormone receptors; predictive model; discriminant analysis; SIMCA
Noninvasive diagnostics of bladder cancer based on analysis of telomerase activity and expression hTERT and hTR coding its subunits by A. I. Glukhov; Y. E. Grigorieva; S. A. Gordeev; A. Z. Vinarov; N. V. Potoldykova (59-68).
In order to develop noninvasive diagnostics of bladder cancer (BC), telomerase activity has been examined by means the TRAP method (telomerase repeat amplification protocol) in tumor tissue and urine pellet samples taken from patients with bladder cancer. The levels of relative expression of genes encoding telomerase catalytic subunit (hTERT) and its RNA subunit (hTR) were evaluated by RT-PCR. Telomerase activity and expression of genes encoding its subunits were detected in both tumor tissues and in the urine cell pellet from each BC patient. Results of our study demonstrate possibility of noninvasive BC diagnostics using combination of these methods with sensitivity of 96% and specificity of 100% in the case of telomerase detection and with sensitivity of 80% and specificity of 100% in the case of hTERT detection in urine pellet samples.
Keywords: bladder cancer (BC); telomerase; telomerase catalytic subunit (hTERT); telomerase RNA-subunit (hTR); diagnositics; noninvasive diagnostics
Highly active fractions of the medicinal leech recombinant destabilase-lysozyme by Yu. I. Fadeeva; N. V. Antipova; I. P. Baskova; L. L. Zavalova (69-72).
Fractionation of the highly purified but low active recombinant protein destabilase-lysozyme (Dest-Lys) by cation exchange chromatography on a TSK CM 3-SW chromatography the non-active fraction (IV) containing 90% of total protein has been separated. Fractions I, II, and III contained proteins with lysozyme and isopeptidase activities and their lysozyme activity correlated with the activity of native Dest-Lys. However, the ratio of lysozyme and isopeptidase activities differed in these fractions; maximal lysozyme activity was found in fraction III, while maximal isopeptidase activity was associated with fraction I. Possible regulation of different functions of Dest-Lys is discussed in the context of formation of its various complexes.
Keywords: medicinal leech; destabilase-lysozyme; lysozyme and isopeptidase functions; forms of a recombinant enzyme
Oxidative stress in the alcoholic liver disease by L. F. Panchenko; B. V. Davydov; N. N. Terebilina; V. Yu. Baronets; A. S. Zhuravleva (73-76).
Parameters reflecting oxidative stress (OS) have been studied in 37 patients with alcoholic liver disease (ALD) during admission to the hospital and 2 weeks after the beginning of therapy. The patients were divided into 3 groups: alcoholic hepatitis (AH), alcoholic cirrhosis with hepatic insufficiency (the group C by the Child-Paquet scale) and terminal stage patients (subsequently died). All patients were characterized by a significant increase in plasma products of lipid peroxidation (conjugated diene and malondialdehyde) and a decrease of the ceruloplasmin level. The coefficient C OS significantly exceeded normal values both on admission and after the 2-week course of traditional therapy. This suggests an important role of the OS with ALD.
Keywords: reactive oxygen species; oxidative stress; alcoholic liver disease; lipid peroxidation; antioxidant system
The effect of doxorubicin incorporated into a phospholipid delivery nanosystem on HepG2 cells proteome by K. G. Kuznetzova; E. V. Kazlas; T. I. Torkhovskaya; P. A. Karalkin; I. V. Vachrushev; T. S. Zakharova; M. A. Sanzhakov; S. A. Moshkovskii; O. M. Ipatova (77-83).
A phospholipid drug delivery nanosystem with particle size up to 30 nm elaborated at the Orekhovich Institute of Biomedical Chemistry (Russian Academy of Medical Sciences) has been used earlier for incorporation of doxorubicin (Doxolip). This system demonstrated higher antitumor effect in vivo as compared with free doxorubicin. In this study the effect of this nanosystem containing doxorubicin on HepG2 cell proteome has been investigated. Cells were incubated in a medium containing phospholipid nanoparticles (0.5 μg/mL doxorubicin, 10 μg/mL phosphatidylcholine). After incubation for 48 h their survival represented 10% as compared with untreated cells. Cell proteins were analyzed by quantitative two-dimensional gel electrophoresis followed by identification of differentially expressed proteins with MALDI-TOF mass spectrometry. The phospholipid transport nanosystem itself insignificantly influenced the cell proteome thus confirming previous data on its safety. Doxorubicin, as both free substance and Doxolip (i.e., included into phospholipid nanoparticles) induced changes in expression of 28 proteins. Among these proteins only four of them demonstrated different in response to the effect of the free drug substance and Doxolip. Doxolip exhibited a more pronounced effect on expression of certain proteins; the latter indirectly implies increased penetration of the drug substance (included into nanoparticles) into the tumor cells. Increased antitumor activity of doxorubicin included into phospholipid nanoparticles may be associated with more active increase of specific protein expression.
Keywords: phospholipid nanoparticles; proteome; doxorubicin; HepG2 cells; MALDI-TOF mass spectrometry