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© 2017, Springer Science+Business Media New York.Hepatoprotective properties of a new pyrimidine derivative — L-ascorbate 1-(2-hydroxyethyl)-4,6-dimethyl-1,2-dihydropyrimidine-2-one, synthesized on the basis Xymedon, were assessed in white rats exposed to CCl4. The compound under study administered prior to exposure to CCl4 reduced the deviation of biochemical parameters from reference values and severity of structural and morphological changes in liver, when compared to the control. Hepatoprotective properties of the studied compound were more pronounced than those of Xymedon.

© 2017 Khaibullin, Ivanova, Martynova, Cherepnev, Khabirov, Granatov, Rizvanov and Khaiboullina.Multiple sclerosis (MS) is an autoimmune neurodegenerative disease characterized by chronic brain inflammation. Leukocyte infiltration of brain tissue causes inflammation, demyelination, and the subsequent formation of sclerotic plaques, which are a hallmark of MS. Activation of proinflammatory cytokines is essential for regulation of lymphocyte migration across the blood-brain barrier. We demonstrate increased levels of many cytokines, including IL-2RA, CCL5, CCL11, MIF, CXCL1, CXCL10, IFNγ, SCF, and TRAIL, were upregulated in cerebrospinal fluid (CSF), whereas IL-17, CCL2, CCL3, CCL4, and IL-12(p40) were activated in MS serum. Interaction analysis of cytokines in CSF demonstrated a connection between IFNγ and CCL5 as well as MIF. Many cells can contribute to production of these cytokines including CD8 and Th1 lymphocytes and astrocytes. Therefore, we suggest that IFNγ released by Th1 lymphocytes can activate astrocytes, which then produce chemoattractants, including CCL5 and MIF. These chemokines promote an inflammatory milieu and interact with multiple chemokines including CCL27 and CXCL1. Of special note, upregulation of CCL27 was found in CSF of MS cases. This observation is the first to demonstrate CCL27 as a potential contributor of brain pathology in MS. Our data suggest that CCL27 may be involved in activation and migration of autoreactive encephalitogenic immune effectors in the brain. Further, our data support the role of Th1 lymphocytes in the pathogenesis of brain inflammation in MS, with several cytokines playing a central role.

© 2017, Pleiades Publishing, Ltd. Based on the active ingredient of the drug Ximedon (1,2-dihydro-4,6-dimethyl-1-N-(2-hydroxyethyl)pyrimidone-2, referred below to as pyrimidine (I), novel derivatives containing biogenic acids: succinic, L-ascorbic, para-aminobenzoic, nicotinic, and L-2-amino-4-(methylthio)butanoic (L-methionine) acids have been synthesized. The parameters of acute toxicity (LD 50 ) have been studied. The antitoxic effect of the compounds upon the injury by the hepatotropic poison carbon tetrachloride has been examined as the primary evaluation of their hepatoprotective properties. It has been found that, according to toxicological safety, the compounds synthesized belong to classes III and IV (moderately and little toxic compounds). The conjugates of pyrimidine (I) with ascorbic acid and methionine (LD 50 more than 5400 mg/kg) are least toxic. Pyrimidine (I) and its derivatives possess the antitoxic activity upon acute poisoning with carbon tetrachloride; the combined injection of carbon tetrachloride with pyrimidine (I) or its derivatives leads to an increase in the survival of animals and the normalization of the integral functional parameters, weight and body temperature, which decrease upon toxic injury. In addition, pyrimidine (I) and some of its derivatives (conjugates with L-ascorbic, succinic, para-aminobenzoic, and nicotinic acids) decrease the weight coefficients of the liver and kidneys (the organ-to-body-weight ratio) and the activity of transaminases, the markers of hepatic cytolysis, which increase upon toxic injury with carbon tetrachloride. The area of the pathological injury of the liver by steatosis and necrosis decreases by the action of pyrimidine (I) and its novel derivatives (conjugates with L-ascorbic, succinic, and nicotinic acids) two to three times. Advantages of pyrimidine (I) and its novel derivatives over the hepatoprotective drug Thiotriazolin have been revealed.

A comparative study of the physical adsorption of RNAse A and RNAse Bacillus pumilis onto a negatively charged surface of mica, a hydrophobic surface of pyrolytic graphite and a surface of lipid layers of dipalmitoylphosphatidylcholine (DPPC) was performed by atomic force microscopy. It was found that microbial RNAse, unlike RNAse A, 1) is adsorbed onto the negatively charged surface of mica in the form of monomers and dimers; 2) exhibits enhanced tropism to the hydrophobic surface of pyrolytic graphite; 3) modifies morphotopography and thickness of the lipid bilayer of DPPC. The detected surface-dependent differences in adsorption of RNAses are consistent with the features of their structure and cytotoxic properties.

© 2016 Khaiboullina, Abdulkhakov, Khalikova, Safina, Martynova, Davidyuk, Khuzin, Faizullina, Lombardi, Cherepnev and Rizvanov.Gastroduodenitis caused by H. pylori, often acquired in early childhood, is found in about 50% of the adult population. Although H. pylori infections can remain asymptomatic, its virulence factors usually trigger epithelial vacuolization and degeneration, loss of microvilli, disintegration of cytoplasm, and leukocyte accumulation. It is believed that leukocyte infiltration is driven by cytokines produced locally in infected tissue. However, so far little is known about changes in serum cytokines in juvenile patients infected with H. pylori. Serum cytokine profiles were analyzed in 62 juvenile patients diagnosed with gastroduodenitis using the Bio-Plex multiplex assay. H. pylori infection was confirmed in 32 patients, while 30 patients were H. pylori-free. Cytokines CXCL5 and CXCL6, potent neutrophil chemoattractants, were upregulated in all patients diagnosed with gastroduodenitis. Serum levels of IL8, a prototype neutrophil attractant, remained unchanged in subjects with gastroduodenitis relative to controls. Therefore, our data suggest that CXCL5 and CXCL6 play a role in directing neutrophil trafficking into inflamed gastroduodenal tissue. In addition, the CCL25/GM-CSF ratio differed significantly between H. pylori-positive and -negative juveniles. Further, study is needed to evaluate the role of CCL25 and GM-CSF in the pathogenesis of the different etiologies of gastroduodenitis.

Transmembrane potential of mitochondria is a sensitive biomarker of metabolic activity of cells. Here, we studied mitochondrial potential Ψm in Yarrowia lipolytica yeast cells treated with two fullerene C60 derivatives: bis-nitroxide methanofullerene and 3-phospho-pentafullerene acid. Transmembrane mitochondrial potential was measured by vital ratiometric cationic fluorochrome JC-1 using flow cytometry. The fullerene C60 derivatives tested in a concentration of 10 μg/ml developed cytoprotective effect in the yeast cells challenged either with non-ionic detergent tween-80, or Tris-buffer, pH 9.0. Treatment with bis-nitroxide methanofullerene resulted in a 6-fold increase in proportion of cells with high Ψm, while 3-phospho-pentafullerene acid evoked a 1,5-fold increase in this subset compared to the stressed cells. Hence, both fullerene derivatives counteract Ψm dissipation in challenged cells.

© 2016 Ekaterina Vasilyeva et al.Crohn's disease (CD) is a chronic inflammatory bowel disease that can be diagnosed at any age. There are two major patient groups based on diagnosis of this disease, before or after the age of 20 (juvenile/adolescent or adult), with disease progression in adults usually milder than in juvenile CD patients. Immune mechanisms have been suggested to play an important role in CD pathogenesis, with cytokines governing the development of the immune response. Upregulation of inflammatory cytokines in serum of juvenile and adult CD patients has been documented; still little is known about age-dependent differences in serum cytokine profiles of CD patients. We applied multiplex technology to analyze serum levels of 12 cytokines in juveniles and adults. We show that during the acute stage of the disease all CD patients have high serum levels of CXCL10, which remains upregulated during remission. Increased serum levels of TNF-α and IL-6 during the acute stage was characteristic of juvenile CD patients, whereas adult CD patients had upregulated levels of GM-CSF and IFN-γ. Taken together, these results demonstrate age-dependent differences in cytokine profiles, which may affect the pathogenesis of CD in patients at different ages of disease onset.

The exact mechanism by which cytotoxic ribonucleases reach the cytosol of tumor cells remains unclear. The interaction of ribonucleases with a lipid bilayer is involved in the translocation of ribonucleases across the endosomal membrane. Here, we aimed to study the hydropathy character of toxic antitumor ribonucleases (bovine seminal ribonuclease and binase) and two non-toxic ribonucleases (bovine pancreatic ribonuclease and human pancreatic ribonuclease) by sliding-window hydrophobicity analysis. Comparative hydropathy plot analysis of the non-toxic pancreatic ribonucleases and their toxic variants was also performed. The data obtained indicate that some cytotoxic ribonucleases have a hydrophobic segment, which is sterically available for the hydrophobic interaction with a tumor cell membrane and endosomal membrane. After dissociation, subunits of dimeric ribonucleases are probably capable of thermodynamically favorable interaction with the interfacial region of a lipid bilayer. Remarkably the hydrophobic segment is not identified in the amino acid sequences of non-toxic ribonucleases. The paper describes the hydrophobic properties of toxic RNases that are essential for both the model of a lipid-protein interaction and the cytotoxicity mechanism unraveling. © 2013 Elsevier Ltd.

The aim of our work was to assess long-term results of autologous peripheral blood hematopoietic stem cell application in patients with peripheral arterial diseases. Peripheral blood hematopoietic stem cells mobilized by granulocyte colony-stimulating factor were transplanted intramuscularly to 30 patients with peripheral arterial diseases (IIb stage by Pokrovsky). Standart tredmill test, anklebrachial index estimation and ankle-brachial index restoration time estimation after loading were performed on 0, 3, 6, 12 and 60 months after transplantation. Immunohistochemical study of injured gastrocnemius muscle biopsies taken before peripheral blood hematopoietic stem cells transplantation and 3 months after the procedure was performed. Peripheral blood hematopoietic stem cells transplantation increases capillary density (22,4%, p = 0,0005). Anklebrachial index increased by 18,1% on month 6 after transplantation without a tendency to change on month 12. 60 months after transplantation initial to hematopoietic stem cells transplantation ankle-brachial index rates were marked. Painless walking distance was increasing at all times of observation progressively, on month 60 no walking distance limitation was marked by most patients. Ankle-brachial index restoring time shows positive trend of the functional state of limb during the first year after transplantation, 60 months after transplantation it showed no «walking reserve» limitation in most patients. 5-years survival was 79%, death causes were stroke, cardiac pathology (3 cases), lung cancer. So, peripheral blood hematopoietic stem cells transplantation allows eliminating peripheral arterial diseases symptoms and preserving limb in long-term period. Autologous transplantation of peripheral blood hematopoietic stem cells has no complications and is safe for therapy of patients with peripheral arterial diseases II stage. © Human stem cells institute, 2013.

Effect of binase (RNAse of Bacillus intermedius) on phorbol myristate acetate-(PMA)-induced apoptosis of human peripheral blood granulocytes and monocytes was studied in vitro by flow cytometry. Both toxic (400 μg/ml) and nontoxic (40 μg/ml) binase concentrations were tested. The binase end-point effect was dependent on the target cell population and the binase concentration. In a granulocyte subset, the 400 μg/ml concentration resulted in strongly pronounced stimulation of PMA-induced apoptosis. In a monocyte subset, the 40 μg/ml concentration developed a protective effect as judged by an increase in a percantage of viable cell subset and by slowing-down cells transtion from an early to late PMA-induced apoptotic phase.

Einleitung: Da das Pankreaskarzinom mit einem deutlich erhöhten Thromboserisiko assoziiert ist, erfolgt unter stationären Bedingungen eine medikamentöse Antikoagulation. Allerdings ist ungeklärt, inwieweit eine regelmäßige Heparinisierung das Tumorwachstum beim Pankreaskarzinom[for full text, please go to the a.m. URL], 126. Kongress der Deutschen Gesellschaft für Chirurgie

Despite the success of antivirals in preventing clinically overt CMV disease in cardiac allograft recipients, sub-clinical active CMV infection remains a major concern because of its association with allograft rejection and vasculopathy. The measurement of CMV specific T-cell responses is a promising approach to assessing this situation. For simplicity, class-1 MHC/peptide-multimers staining CD8 T-cells directly are often used but this ignores a much wider range of responses including the whole CD4 T-cell compartment. CD4 T-cells, however, were recently shown to be critical to reducing CMV load early after transplantation. To determine how extensive T-cell responses to CMV are, the responses to two dominant CMV proteins, IE-1 and pp65, were dissected in detail accounting for T-cell lineage, frequencies, epitope recognition and changes over time in more than 25 heart transplant recipients. Cross-sectional results from over 30 healthy CMV-carriers were analyzed for comparison. Responses were unexpectedly complex, with considerable inter-individual variation in terms of dominance, breadth, and recognized epitopes. Whereas the use of MHC/peptide-multimers for clinical CD8 T-cell response monitoring alone can be justified in some situations, short term T-cell activation combined with intracellular cytokine staining was clearly found to be of more general usefulness. The performance of IFN-gamma, TNF-alpha, or IL-2 as single read-outs in identifying activated T-cells was examined and confirmed that the frequently used IFN-gamma was best suited. These results should be used to inform the design of clinically applicable and diagnostically useful approaches to monitoring CMV specific responses in heart transplant recipients.

The magnitude of transmembrane potential Δψ in cells of Escherichia coli K12 was determined by the method of flow cytofluorometry for different phases of growth. It was large in the log phase, whereas in the lag and stationary phases, the population was shown to consist of two subpopulations with low and large values of Δψ in cells. In the presence of 200 mg/l of 2,4,6-trinitrotoluene (TNT), this bimodal distribution of Δψ over the population was observed during the entire growth period until TNT was almost completely eliminated from the cultivation medium (to a concentration of 18-20 mg/l). The mean value of Δψ in cells of the population grown in the presence of TNT was substantially smaller than that in controls due to the larger fraction of the subpopulation with a low value of Δψ. Upon elimination of TNT, the distribution of Δψ in cells of the culture became unimodal and close to that in the control culture in the early log phase of growth. These findings are discussed from the standpoint that considers heterogeneity of the culture of Escherichia coli K12 as a mechanism of its adaptation to the presence of xenobiotics. © Nauka/Interperiodica 2007.

The hydroxystilbene trans-3,5,3,4-tetrahydroxystilbene (piceatannol) (1), isolated from the methanol extract of Euphorbia lagascae defatted seeds, was methylated to yield the derivatives trans-3,5,3,4'-tetramethoxystilbene (2), (trans-3,5-dihydroxy-3', 4'-dimethoxystilbene) (3) and trans-3,5,3'-trihydroxy4'-methoxystilbene (4). The structures of the compounds were assigned by spectroscopic methods (IR, H-1-NMR, C-13-NMR and MS). The ability of piceatannol (1) and the three methylated derivatives to modulate the transport activity of P-glycoprotein (P-gp) and apoptosis induction on the L5178 mouse lymphoma cell line containing the human MDR1 gene was studied by flow cytometry. The reversal of multidrug-resistance (MDR) was investigated by measuring the accumulation of rhodamine-123, a fluorescent substrate analog of doxorubicin, in cancer cells. Verapamil was applied as a positive control. For the evaluation of the compounds as apoptosis inducers, tumor cells were stained with FITC-labelled annexin-V and propidium iodide. The tetramethylated derivative (2) was found to be a powerful inhibitor of P-gp activity. Compounds I and 2 showed an increased apoptotic effect in the MDR subline, the most active being piceatannol (1). Furthermore, in the combination chemotherapy model, the interaction between doxorubicin and the resistance modifier 2 was studied in vitro. The results of checkerboard experiments indicated that the type of interaction was additive between doxorubicin and compound 2 on the human MDR1 gene-transfected mouse lymphoma cells. However, in the MCF7/dox human breast cancer cells, the interaction was non-additive. The degree of additive and non-additive interactions were close to the borderline of the FIX values corresponding to the two types of interactions.

An induction of apoptosis by RNase from Bacillus intermedius (binase) and its mutants characterized with low catalytic activity (Lys 26Ala and His 101Glu) in human myelogenic erythroleukemia K562 cells, human lung carcinoma A549 cells and human peripheral blood mononuclear cells was studied. For the first time selective apoptogenic effects of binase toward leukemic blood cells was determined. Neither antiproliferative nor apoptotic effects of binase were detected in normal human peripheral blood mononuclear cells. Formation of low molecular weight oligonucleosomal DNA fragments (less than 50 Kb) which are an early marks of apoptosis was registered in solid tumor cells treated by binase. Using mutant RNases it was shown that decrease of catalytic activity to 2.5% of wild type enzyme activity leads to the loss of apoptogenic properties of enzyme. Selective apoptogenicity of binase found towards malignant cells confirmed that antitumor agents based on bacterial RNases could be considered as an alternative to standard chemotherapeutic drugs.