The antimicrobial activity and cytotoxicity of novel 1,3-bis(alkyl)-6-methyluracil derivatives containing 1,2,3- and 1,2,4-triazolium fragments in alkyl chains have been studied. The compounds have been tested for the antimicrobial activity toward some gram-positive and gram-negative bacteria and fungal cultures. The cytotoxic action has been estimated toward mammalian cells. It has been found that the basic structural factor that affects the antimicrobial activity is the nature of alkyl radicals at triazole fragments.
A series of semicarbazones, thiocarbazones, 1,3,4-oxadiazoles, and 1,3,4-thiadiazoles bearing coumarin and pyrazole moiety have been synthesized. The new synthesized compounds were screened in vitro for their antimicrobial and antioxidant activities. Preliminary studies showed that among the synthesized new compounds, chloro-substituted thiosemicarbazone showed excellent activities against all tested organisms; at the same time, methyl substituted thiosemicarbazone showed greater activity against E. coli. Chloro-substituted 1,3,4-oxadiazole and 1,3,4-thiadiazole demonstrated greater DPPH and hydroxyl radical scavenging abilities. Molecular docking studies indicate that 1,3,4-oxadiazoles and 1,3,4-thiadiazoles manifest better interaction with CAT (catalase) and GPx (glutathione peroxidase) than that with SOD (superoxide dismutase). Studies on the antimicrobial and antioxidant activities of the synthesized compounds compared with those of their starting compounds are discussed.
Some novel bicyclic thiazolopyrimidine derivatives bearing various substituents have been synthesized through one-pot three-component method. Structures of the target compounds were confirmed by elemental analysis and spectral data. Some selected members of the newly synthesized compounds were investigated for their analgesic and anti-inflammatory activities and revealed pronounced anti-inflammatory activity greater than that of indomethacin (reference drug).
A proteomic analysis of the venom of males and females of the Naja kaouthia monocled cobra specimens kept in captivity was carried out. Using the amino acid sequences of proteins of the taxonomic group Serpentes from the UniProt KB database, 875 proteins were identified in the venom samples and the relative content of about 190 of them was determined in each venom sample by the method of label-free quantitative proteomics. The total content of 35 major protein components of the venom of each individual was shown to comprise about 98% of the total amount of venom proteins. Analysis of relative content of the venom proteins in males and females showed a significantly (p < 0.05) higher content of nerve growth factor and natriuretic peptide in the venom of females. Analysis of the profile of posttranslational modifications of N. kaouthia toxins revealed previously unknown sites for phosphorylation, acetylation, and formylation.
Polypeptide SE-33 (SETRPVLNRLFDKIRQVIRKFEKGIKEKSKRFF), which is a retro analog of natural antimicrobial protein cathelicidin LL-37 (LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES), was synthesized by the method of solid-phase peptide synthesis. Similar to the natural peptide, polypeptide SE-33 forms an amphipathic alpha helix but has an inverted amino acid sequence compared to cathelicidin. It has been shown the physicochemical properties of polypeptide SE-33 are similar to those of the natural compound. In vitro experiments have shown that polypeptide SE-33 exerts a bactericidal effect on the cells of Staphylococcus aureus Wood 46, which is comparable with the effect of cathelicidin LL-37, as well as pronounced antifungal activity against the clinical isolates of Candida albicans, Cryptococcus neoformans, Rhodotorula mucilaginosa, Trichosporon cutaneum, Geotrichum sp. The MICs of polypeptide SE-33 for different fungal strains were in the range of 31.2 to 1024 μg/mL. Polypeptide SE-33 demonstrated high activity in vivo in the model of vulvovaginal candidiasis (VVC) in mice comparable with that of pimafucin. In the absence of side effects and signs of pathology, polypeptide SE-33 in all doses tested (1, 10 and 50 mg/mL) statistically reduced the vaginal load of the mice compared to the placebo group. The pronounced antibacterial and antifungal activity of polypeptide SE-33, as well as the absence of a toxic effect in the VVC model in mice, suggest polypeptide SE-33 as a promising antimicrobial agent.
Different methods of covalent lysozyme immobilization have been compared to choose an optimal approach to the development of a material suitable for medical applications in extracorporeal therapy. A novel method for lysozyme immobilization on a polymeric agarose matrix has been proposed, which provides the effective action of lysozyme on bacterial cells and eliminates the leakage of the enzyme from the material. The resultant immobilized lysozyme exhibits bacteriolytic activity toward Gram-positive bacteria Micrococcus luteus and Gram-negative bacteria Escherichia coli. During the immobilization of the enzyme, a broadening of the pH optimum of its activity occurs. The compatibility of immobilized lysozyme with human whole blood has been shown.
MicroRNAs (miRNAs) are a family of ∼22-nucleotide, non-coding, single-stranded RNA molecules, which are considered as key regulators of gene expression and regulate various biological processes including gonad development, differentiation and dimorphism. Although most miRNAs studied have been identified in animals, relatively limited knowledge also exist on amphibian species. Identification of the full repertoire of miRNAs in testes of Andrias davidianus would significantly increase our knowledge on miRNA physiological functions of miRNA in gonads. In this study, a small RNA library was constructed from adult A. davidianus testis tissue and sequenced using the deep sequencing. As a result, 26 401 474 clean reads was obtained, and 1340 known conserved miRNAs were identified. Additionally, expression of 12 randomly selected miRNAs was confirmed by stem-loop qRT-PCR from 5 different tissues of A. davidianus. Of them, aca-miR-99b-5p, xtr-miR-10c, and cfa-miR-363 were highly expressed in testes. Although the physiological functions of these expressed miRNAs were still not revealed, their expression patterns indicate that they are likely to play a role in growth and development of A. davidianus testes, which is needed to be further study for confirmation. The results of this study will help in gaining understanding of the role of miRNAs in the regulation of A. davidianus reproductive system.
The role of nAChRs in functioning of central and peripheral nervous systems is widely known. Recently, expression of nAChRs in immune cells and tissues of epithelial origin has been shown. These non-neuronal nAChRs are involved in regulation of inflammation, cell proliferation, differentiation, migration, and apoptosis. Non-neuronal nAChRs are also involved in occurrence and development of several types of nicotine-induced cancers. Study of these receptors in epithelial and cancer cells made it possible to discover new ways of nAChRs regulation by endogenous ligands and to propose new mechanisms of signal transduction that are not related to the ion current through the receptor channel. This review presents current information on the function of human non-neuronal nAChRs.
The early period of postnatal development of premature infants is often complicated by bacterial infections, in particular respiratory infections, which in adverse course can be transformed into generalized forms (sepsis), leading to life-threatening conditions. The development of these complications may be associated with delayed diagnosis and delayed start of targeted therapy. Thus, the development of new approaches to the diagnosis of inflammatory pathological processes, especially in the first hours of life, is an important area of research in modern neonatology. In this work the profiling of blood plasma lipids from newborn premature babies was performed in order to identify potential markers of inflammatory processes. The study involved three groups of patients who were diagnosed with pneumonia, pneumonia complicated by sepsis, or respiratory distress syndrome in the early postnatal period. As a result of HPLC-MS analysis of blood plasma lipid extracts, characteristic molecular profiles were obtained, which revealed significant differences between the groups under study. Analysis of the molecular composition showed differential representation of lipid classes such as phosphatidylcholines, phosphatidylethanolamines, di- and triglycerides, sphingolipids and lysophospholipids. Our results may indicate that the inflammatory processes at the local and systemic levels affect lipid metabolism and their composition in blood plasma. Moreover, each pathology—sepsis, pneumonia or RDS—has its own specific lipid profile, which may be useful not only to detect inflammation but also to differentiate inflammation-associated diseases from each other.
A radioligand-based method was proposed for quantifying the binding activity of [beta]1-adrenergic receptors (ARs) on the surface of human T lymphocytes. Using the transfected HEK293 cell lines expressing [beta]1-AR (ADL-7A) and [beta]2-AR (A2R9) as a model, the conditions for [[.sup.125]I]cyanopindolol reliable detection of specific ligand binding to [beta]1-adrenergic receptors at the level of 11.5 fmol per 1 million cells were selected. The real possibility of performing this analysis for clinical studies was shown by the example of human T lymphocytes.
Diketone DNA derivatives have been proposed to modify the guanidine group of Arg in proteins. The [beta]-diketo group at the C2' atom of the sugar phosphate moiety has been introduced in DNA by acylation of oligonucleotide precursors, i.e., DNA fragments containing 2'-amino-2'-deoxyuridine, which have been synthesized by the chemical automatic synthesis. Water-soluble N-[3-(dimethylamino)propyl]-N -ethylcarbodiimide (EDC) and 4,6-dioxoheptanoic acid have been used in the reaction. The ability of oligodeoxyribonucleotides containing the 2'-[beta]-diketo group to react with guanidine, N.sup.[alpha]-Boc-L-arginine, and N.sup.[alpha]-Dns-L-arginine has been demonstrated. The introduction of this modification into one of the strands of the 15-base pair DNA duplex has been shown to lead to its destabilization. The conjugate formation of MutS and MutL proteins from the E. coli mismatch repair system with 17-base pair DNA duplexes containing the 2'-deoxy-2'-(4,6-dioxoheptylamido)uridine residue has been detected for the first time. To increase the selectivity of the DNA ligands containing the [beta]-diketo group in the reaction with the Arg residues of proteins, we have proposed to treat the reaction mixture with hydroxylamine. This treatment leads to the cleavage of Schiff bases, which are formed with the involvement of lysine residues. (4) 0000 0001 0482 5067, grid.34980.36, Department of Biochemistry, Indian Institute of Science, 560012, Bangalore, India Corresponding author: phone: +7 (495) 939-54-11 fax: +7 (495) 939-31-81 e-mail: firstname.lastname@example.org
Diketone DNA derivatives have been proposed to modify the guanidine group of Arg in proteins. The β-diketo group at the C2' atom of the sugar phosphate moiety has been introduced in DNA by acylation of oligonucleotide precursors, i.e., DNA fragments containing 2'-amino-2'-deoxyuridine, which have been synthesized by the chemical automatic synthesis. Water-soluble N-[3-(dimethylamino)propyl]-N′-ethylcarbodiimide (EDC) and 4,6-dioxoheptanoic acid have been used in the reaction. The ability of oligodeoxyribonucleotides containing the 2'-β-diketo group to react with guanidine, Nα-Boc-L-arginine, and Nα-Dns-L-arginine has been demonstrated. The introduction of this modification into one of the strands of the 15-base pair DNA duplex has been shown to lead to its destabilization. The conjugate formation of MutS and MutL proteins from the E. coli mismatch repair system with 17-base pair DNA duplexes containing the 2'-deoxy-2'-(4,6-dioxoheptylamido)uridine residue has been detected for the first time. To increase the selectivity of the DNA ligands containing the β-diketo group in the reaction with the Arg residues of proteins, we have proposed to treat the reaction mixture with hydroxylamine. This treatment leads to the cleavage of Schiff bases, which are formed with the involvement of lysine residues.
One of the means for regulating the plant innate immune system is activating the synthesis of various defense peptides having diverse structural organization. Some of them possess antimicrobial activity. In plants, defense peptides have various localization; they are often synthesized as multidomain precursor proteins or produced by limited proteolysis, as well as through other degradation pathways. In addition to antimicrobial activity, some of these peptides also display an insecticidal effect, inhibit endo- and exogenous proteases, as well as α-amylases, participate in the transfer of building and signaling hydrophobic molecules, and affect the functioning of ion channels. As a result, peptides of plant innate immune systems not only protect plants against viruses, bacteria, fungi, and insects, but also reinforce their resistance to various types of abiotic stresses and participate in the regulation of plant growth and development. In addition, plant defense peptides can suppress the growth of some human pathogens, possess antitumor activity, exhibit properties of food, inhalation, and latex allergens, and can be used in various areas of medicine. This review summarizes data on biosynthesis, biological functions, and possible practical application of the peptides of the plant innate immune system.
A radioligand-based method was proposed for quantifying the binding activity of β1-adrenergic receptors (ARs) on the surface of human T lymphocytes. Using the transfected HEK293 cell lines expressing β1-AR (ADL-7A) and β2-AR (A2R9) as a model, the conditions for [125I]cyanopindolol reliable detection of specific ligand binding to β1-adrenergic receptors at the level of 1–1.5 fmol per 1 million cells were selected. The real possibility of performing this analysis for clinical studies was shown by the example of human T lymphocytes.
Current studies of fungal bioluminescent systems, including the purification of luciferase, require large quantities of the synthetic substrate luciferin available. The existing synthetic method for the luciferin produces low yield of the target molecule. In the current work, the synthesis of fungal luciferin was scaled up to gram quantities and an effective method for its final purification was found. The total yield in comparison with the previous method was increased by 3.5 times; the overall cost of synthesis fell by 3 times.
Lasting contact of medical personnel in tuberculosis departments with the pathogen of Mycobacterium tuberculosis causes changes in their immune system, which is aimed at adapting the body to a constant antigenic load. Changes in the blood of medical staff indicate the participation of certain cells in the formation of protective immunity. Therefore, the observed changes are likely to be specific and reflect the formation of the protection of a healthy person’s organism from the development of tuberculosis. The question of the relevancy of the use of medical personnel blood parameters as a control over the health status of chronic tuberculosis patients is discussed. The most pronounced changes were observed in populations of monocytes and CD4+ T cells, which are directly involved in the protection of the human body against tuberculosis. The number of CD4+ T cells capable of producing interferon-gamma (IFN-γ) in response to stimulation by M. tuberculosis antigens was analyzed in the blood of medical personnel. The number of these cells in the blood of the medical staff varied depending on the duration of their work in clinic. Low level of CD4+ (IFN-γ)+ activated T cells in the blood of employees after 5–7 years of work can be a prognostic factor for low resistance to tuberculosis.
The insulin receptor-related receptor (IRR) is a cellular sensor of a weakly alkaline medium. Its spatial structure and the mechanism of activation have not been yet established. In the present work, a system of heterologous expression of full-length IRR tyrosine kinase has been created. A plasmid construct encoding the cDNA of the full-length human IRR fused at the C-terminus with the green fluorescent protein (GFP) has been generated, and the hybrid protein has been expressed in HEK293 line cells. It has been shown that the receptor incorporated in the hybrid protein is expressed on the cytoplasmic membrane and retains its functional activity. The resulting protein can be used in further studies of the structure and function of IRR.
The review is devoted to promising directions of the search for chemical compounds which can be used as the basis for the development of new drugs for migraine treatment. It considers promising biological targets, whose effects are capable of eliminating the manifestations of acute migraine attacks, and also indicates the main existing developments in the form of chemical compounds and drug prototypes interacting with these targets, including those that will soon enter or are at the stage of clinical trials. In particular, preclinical and clinical data on compounds whose effects are determined by the influence on the calcitonin gene-related peptide (CGRP) receptors, serotonin 5-HT.sub.2A and 5-HT.sub.1F receptors, NO-synthase, orexin and glutamate receptors, are discussed. The review focuses on the description of 5-HT.sub.2A receptors as a promising target for the development of novel antimigraine drugs. Information about the structural features of this serotonin receptor subtype, the localization and functions is given also it involves the pharmacological capabilities of new substances affecting 5-HT.sub.2A receptors. The final part of the review is devoted to new trends in the search for 5-HT.sub.2A antagonists among compounds synthesized based on a serotonin bioisostere, benzimidazole.
Conjugates of 3,4,6-tri-О-acetyl-N-acetylglucosamine and tetraacetyl glucopyranose with alkyl phosphates were synthesized. The dependence of their antibacterial and antituberculosis activities on the length of the alkyl substituent at the phosphate group was found. The conjugates with a decyl substituent exhibited in vitro the highest antituberculosis activity against Mycobacterium tuberculosis H37Rv (MIC 3 µg/mL) but the weakest effect towards Streptococcus aureus and Bacillus cereus (≤MIC 125 µg/mL). Vice versa, the conjugates with a cetyl substituent demonstrated the highest antibacterial activity in vitro towards S. aureus and B. cereus (MIC 16 µg/mL) but showed the lowest antituberculosis activity (MIC 12 µg/mL) among the compounds under study.