Synthesis of Deuterium-Labeled Pyrrolylcarnosine.

The effect of temperature on the effectiveness of the incorporation of deuterium into pyrrolylcarnosine (PC) was studied. Deuterium gas and heavy water were used as a source of deuterium. Isotope exchange was carried out using solid-phase and liquid-phase methods. It was found that it is better to use isotope exchange with deuterated water to obtain preparative amounts of labeled pyrrolylcarnosine. When using y solid-phase method, the main label is in pyrrole. The incorporation of deuterium at a higher temperature occurs more evenly. In addition, the use of deuterated water made it possible to reduce the amount of unlabeled isotopomer to almost 0% and to obtain a product with a yield of 70% and a content of more than seven deuterium atoms. It was established that the content of deuterium in the compound can be increased by pretreating the reaction mixture with deuterium gas. This approach opens up additional opportunities for the synthesis of labeled compounds.

Assessment of the Cytotoxicity of Peptide Modifications of Doxorubicin on Tetrahymena pyriformis.

The cytotoxicity of doxorubicin (Dox) and its peptide modifications Z-Gly-Pro-Dox and Boc-Gly-Pro-Dox were studied. Tetrahymena pyriformis was used as a test system, which made it possible, due to the short life cycle and high reproduction rate of ciliates, to trace their response to the effects of toxicants over several generations. It was found that peptide modification of the Dox molecule markedly reduces its cytotoxic and cytostatic effect. The Z-Gly-Pro-Dox modification has less cytotoxic and cytostatic effect compared to Boc-Gly-Pro-Dox. When determining the ability of drugs (at a concentration of 100 µM) to prevent bacterial contamination of samples, it was shown that the smallest degree of overgrowth was recorded in the presence of Dox (OD600nm 81.1). Boc-Gly-Pro-Dox also had a bacteriostatic effect, though less pronounced (OD600nm 93.8). The degree of overgrowth in the presence of Z-Gly-Pro-Dox was close to that of distilled water. The results obtained on ciliates did not contradict the data obtained in similar studies on mice.

Efficiency of Penetration of Dopamine and Serotonin Peptide Derivatives through the Artificial Membranes.

A mass spectrometric method has been developed for determining the content of dopamine and serotonin derivatives, which allows evaluating the efficiency of their penetration through artificial membranes depending on the structure of their peptide fragment. In this case, the diffusion of dopamine and serotonin derivatives through the membrane occurred as a result of competitive interactions. It was shown which compounds in this mixture more easily penetrate through artificial membranes. It was found that the most promising in terms of overcoming the BBB are Boc-Pro-Srt and Boc-Pro-DOPA.

[Antegrade approach for cholangiolithiasis complicated by mechanical jaundice]. / Vozmozhnosti antegradnogo dostupa v lechenii kholangiolitiaza, oslozhnennogo sindromom mekhanicheskoi zheltukhi.

AIM: To improve an efficiency of surgical treatment of patients with cholelithiasis complicated by obstructive jaundice through antegrade interventional approach. MATERIAL AND METHODS: 166 patients aged from 23 to 92 years with cholangiolithiasis complicated by mechanical jaundice were enrolled. Patients were divided into 2 groups: group I (136) - retrograde endoscopic method, group II (30) - antegrade interventional approach. RESULTS: In the first group surgical efficacy was 79.4%. Morbidity and mortality were 13% and 2% respectively. In the second group these values were 96.7%, 10% and 3% respectively. CONCLUSION: Antegrade interventional approach for minimally invasive procedures is technically feasible, has the same effectiveness as the retrograde endoscopic method and also all advantages of minimally invasive techniques.

Horizontal Transfer of Tamoxifen Resistance in MCF-7 Cell Derivates: Proteome Study.

Using estrogen-dependent MCF-7 breast cancer cells and tamoxifen-resistant MCF-7/T subline we have shown that their co-cultivation lead to increase in tamoxifen resistance in the parent MCF-7 cells. The proteome analysis of MCF-7/T cells and new-generated resistant cells revealed 21 common proteins differently expressed in both the resistant cell lines, among them - 6 proteins were associated with the drug or hormonal resistance. Both resistant lines were characterized with suppression of estrogen receptor and activation of SNAIL1-signaling - mesenchymal pathway playing an important role in the down-regulation of estrogen receptor and maintaining of the estrogen-independent phenotype.

Is myelin basic protein a potential biomarker of brain cancer?

Myelin basic protein is a potential biomarker for the central nervous system diseases in which the myelin sheath is destroyed. Using pseudo-selected reaction monitoring and the method of standard additions, we have measured the myelin basic protein level in the cerebrospinal fluid of patients with neurotrauma (n = 6), chronic neurodegenerative diseases (n = 2) and brain cancer (n = 5). Myelin basic protein was detected only in four out of five cerebrospinal fluid samples of patients with brain cancer. The cerebrospinal fluid myelin basic protein level ranged from 3.7 to 8.8 ng ml-1. We suggest that monitoring of myelin basic protein in cerebrospinal fluid can serve as a diagnostic test for the brain cancer.

The penetration of 5-oxo-Pro-Arg-Pro into the brain and the major metabolic pathways of this peptide in the rat brain and blood at the intranasal and intravenous administration.

It was shown that the neuroactive peptide 5-oxo-Pro-Arg-Pro (5-oxo-PRP) is detected in the brain in the time interval of 5-120 min after it was intravenously or intranasally administered to rats; the maximum concentration of labeled tripeptide in these modes of administration was observed after 30 and 10 min, respectively. A significant difference in the concentrations of 5-oxo-PRP in the blood and brain (the latter was 50 times lower) during intravenous administration indicates a relatively low permeability of the peptide across the blood-brain barrier. Pharmacokinetic data analysis showed that, when administered intranasally, approximately 45% of the total number of 5-oxo-PRP detectable in the brain in the entire period of study enters via transport from the nasal cavity, and the rest of the peptide enters through the blood-brain barrier from the blood stream. It was found that 5-oxo-PRP in rats is rapidly metabolized forming proteolytic products, mainly amino acids, and degradation products, presumably oxidized these amino acids.

Synthesis of N-acyl derivatives of Pro-Gly-Pro-Leu peptide: Proteolytic stability in vitro and effects on mouse macrophage cells RAW264.7.

Acetyl, oleoyl, arachidonoyl, and docosahexaenoyl derivatives of the Pro-Gly-Pro-Leu peptide with a chemical purity of 99.8% were synthesized. The degradation kinetics of the Pro-Gly-Pro-Leu derivatives under the action of leucine aminopeptidase, nasal mucus, and microsomal fraction of the brain and blood of rats was studied. It was shown that the N-acyl derivatives of Pro-Gly-Pro-Leu proved to be more resistant to the action of leucine aminopeptidase and other enzyme systems. The study of the cytotoxic and anti-inflammatory activity of preparations on the mouse macrophage cell line RAW264.7 showed that acylation with oleic and arachidonic acid makes the peptide cytotoxic with LC50 in the range of 70-15 µM and gives it anti-inflammatory properties with EC50 of 32 and 36 µM, respectively.

[Polymorphisms in Genes of Cytokines and Matrix Metalloproteinases Associated With Ischemic Heart Disease in Patients With Type 2 Diabetes].

OBJECTIVE: To examine associations between ischemic heart disease (IHD) and polymorphisms in cytokine genes (IL-1B, IL-4, IL-6, IL-10, TNFA, VEGF) and matrix metalloproteinase genes (MMP2, MMP3, MMP9) in patients with type 2 diabetes. MATERIAL AND METHODS: We studied 232 Caucasian diabetic subjects (33 men and 199 women aged 50-70 years). In 93 patients IHD was verified by treadmill test and/or coronary angiography (86 subjects with stable angina, 19 with previous myocardial infarction). Thirteen polymorphisms localized in the promoters of IL-1B (rs1143627), IL-4 (rs2243250), IL-6 (rs1800795), IL-10 (rs1800872, rs1800896), TNFA (rs361525, rs1800629, rs1800630), VEGF (rs699947, rs3025039), MMP2 (rs243865), MMP3 (rs3025058) and MMP9 (rs3918242) were investigated. RESULTS: Prevalence of G-allele and GG-genotype at -308 position of TNFA (rs1800629), as well as C-allele and CC-genotype at position +936 of VEGF (rs3025039) was higher in patients with IHD as compared to patients without IHD (OR=2.0, OR=2.2, OR=2.1, OR=2.4, respectively, all p=0.02). In logistic regression analysis, TNFA -308 A/G and VEGF +936 C/T polymorphisms showed associations with IHD (both p=0.009). These polymorphisms along with age, body mass index, duration of diabetes, low density and high density lipoprotein cholesterol were associated with IHD in multivariate models (p=0.0002 and p=0.00008, respectively). Nine combinations of TNFA -308 GG-genotype and variants of other genes demonstrated associations with IHD (p≤0.002). CONCLUSION: The polymorphisms in promoter regions of TNFA (rs1800629) and VEGF (rs3025039) are associated with IHD in patients with type 2 diabetes.

Glutamate release and uptake processes are altered in a new mouse model of amyotrophic lateral sclerosis.

In this paper, we showed that in the cortex of mice expressing an abberant form of FUS protein that model amyotrophic lateral sclerosis (ALS), the processes of KCl-induced and basal [(3)H]glutamate release and uptake are altered at the presymptomatic stage as compared to the non-transgenic littermates. The change in these three parameters in transgenic animals causes excitotoxicity, which, in turn, may lead to massive loss of motor neurons and the onset of ALS symptoms.

Electron kinetics inferred from observations of microwave bursts during edge localized modes in the mega-amp spherical tokamak.

Recent measurements of microwave and x-ray emission during edge localized mode (ELM) activity in tokamak plasmas provide a fresh perspective on ELM physics. It is evident that electron kinetics, which are not incorporated in standard (fluid) models for the instability that drives ELMs, play a key role in the new observations. These effects should be included in future models for ELMs and the ELM cycle. The observed radiative effects paradoxically imply acceleration of electrons parallel to the magnetic field combined with rapid acquisition of perpendicular momentum. It is shown that this paradox can be resolved by the action of the anomalous Doppler instability which enables fast collective radiative relaxation, in the perpendicular direction, of electrons accelerated in the parallel direction by inductive electric fields generated by the initial ELM instability.

Proteolysis of His-Phe-Arg-Trp-Pro-Gly-Pro in the blood and brain of rats in vivo.

The kinetics of the content of His-Phe-Arg-Trp-Pro-Gly-Pro (ACTH (6-9)PGP) and its hydrolysis products in the blood and brain of rats in the case of intranasal administration and intravenous injection of tritiated ACTH(6-9)PGP was studied. The parameters of bioavailability of ACTH(6-9)PGP administered intranasally were higher, indicating certain prospects in the intranasal application in clinical practice. We also found that the factor that determines ACTH(6-9)PGP proteolysis in experiments both in vivo and in vitro is aminopeptidases. The main products of ACTH(6-9)PGP during its metabolism in rats are short peptides and amino acids.

Penetration of thyroliberin in the blood and brain regions at intranasal or intravenous administration.

The maximum amounts of the thyroliberin in the blood and brain of rats at intranasal and intravenous administration were determined. It is found that rat hippocampal, cortical, and cerebellar membranes contain two types of specific binding sites (high- and low-affinity) for the labeled ligand. It was shown that, at intranasal and intravenous administration, maximum amounts of the thyroliberin were detected in the cerebellum and then in the cortex and hippocampus. The degradation of the thyroliberin in the rat brain and its regions at intranasal and intravenous administration was studied. It is shown that the degree of degradation and the formation of proteolytic products of the thyroliberin is different in different regions of the rat brain.

High-resolution structure of a membrane protein transferred from amphipol to a lipidic mesophase.

Amphipols (APols) have become important tools for the stabilization, folding, and in vitro structural and functional studies of membrane proteins (MPs). Direct crystallization of MPs solubilized in APols would be of high importance for structural biology. However, despite considerable efforts, it is still not clear whether MP/APol complexes can form well-ordered crystals suitable for X-ray crystallography. In the present work, we show that an APol-trapped MP can be crystallized in meso. Bacteriorhodopsin (BR) trapped by APol A8-35 was mixed with a lipidic mesophase, and crystallization was induced by adding a precipitant. The crystals diffract beyond 2 Å. The structure of BR was solved to 2 Å and found to be indistinguishable from previous structures obtained after transfer from detergent solutions. We suggest the proposed protocol of in meso crystallization to be generally applicable to APol-trapped MPs.

Proteolysis of Pro-Gly-Pro-Leu in the hippocampus, cerebellum, and cerebral cortex in rats after intravenous injection.

We studied the dependence of the levels of Pro-Gly-Pro-Leu peptide and its metabolites (Gly-Pro-Leu, Pro-Gly-Pro, Gly-Pro and Pro-Gly) from the time of administration in the hippocampus, cerebellum, and cerebral cortex of rats. After intravenous injection of Pro-Gly-Pro-Leu, the maximum concentration of metabolites in the rat brain was found in 20-min sample (0.026% from the amount of introduced labeled tetrapeptide); it was by 2.6 times higher that after intranasal administration. The calculated ratios of the peptide content (AUC) in brain structures for Pro-Gly-Pro-Leu and its metabolites after intranasal and intravenous injection were in most cases >1; hence, the levels of these peptides in the cerebellum, hippocampus, and brain cortex after intravenous injection were slightly higher than after intranasal administration. From these data, content of Pro-Gly-Pro-Leu and its metabolites per 1 g of the cerebellum, hippocampus and cortex was calculated: the maximum concentrations of Pro-Gly-Pro-Leu in the cerebellum, hippocampus, and cortex were 15.63, 18.48, and 2.95 pmol/g, respectively.

Nonstoichiometry as a hidden aspect of TbAl3(BO3)4 optical properties.

TbAl3(BO3)4 should not be considered as a strictly stoichiometric compound. A variety of Tb1+xAl3-x(BO3)4 (x = -0.1-0.15) single phase compositions with the R32 space group were synthesized in the TbBO3-(Al2O3·B2O3) system. The K2Mo3O10-B2O3-Al2O3 flux was used to grow Tb1+xAl3-x(BO3)4 (x = 0.06 and 0.09) crystals. The orthoborates have typical luminescence in the green range which correlated with the 5D3 → 7F6 electron transition of Tb3+. However, deviating from stoichiometry can result in halving the quantum efficiency of luminescence. The SHG efficiency from 1064 nm radiation for Tb1+xAl3-x(BO3)4 crystals (x = 0.06 and 0.09) was found to be 1.82 and 1.53 times higher than that of KDP, respectively.

Induced emission of Alfvén waves in inhomogeneous streaming plasma: implications for solar corona heating and solar wind acceleration.

The results of a self-consistent kinetic model of heating the solar corona and accelerating the fast solar wind are presented for plasma flowing in a nonuniform magnetic field configuration of near-Sun conditions. The model is based on a scale separation between the large transit or inhomogeneity scales and the small dissipation scales. The macroscale instability of the marginally stable particle distribution function compliments the resonant frequency sweeping dissipation of transient Alfvén waves by their induced emission in inhomogeneous streaming plasma that provides enough energy for keeping the plasma temperature decaying not faster than r(-1) in close agreement with in situ heliospheric observations.

[Prostate cancer detection using mass-spectrometric profiling of low-molecular weight blood proteome].

Currently, there are no reliable biomarkers of blood plasma for early detection of prostate cancer (PC), a one of the most common malignancies in men. This study developed, universalized and tested a new standardized methodology of detection of prostate cancer biomarkers--profiling of low-molecular weight proteome of blood plasma (1-17 kDa). This approach includes three main components: a preliminary sample preparation, time-of-flight mass spectrometry with an matrix-assisted laser desorption/ionization ALDI-TOF-MS), and the processing of data using bioinformatics software package. The potentials and prospects of the approach developed for identification of potential PC markers are demonstrated. 46 samples of blood plasma of PC patients and 26 controls were screened. This evaluation identified peptides/polypeptides that have the potential to be used for the detection of disease.

[Functional state of the hemostasis system in patients, operated for pancreatic pseudocysts].

Operative treatment of pancreatic pseudocysts coincides with the indices of the blood coagulation system change, which are characterized as a hypercoagulative. These disorders are noted in 12-24 h postoperatively, their rate is reducing substantially while application of pentoxyphylline. Most significant lowering of intraoperative hypercoagulation was noted while application of endoscopic drainage with the pseudocysts cavity stenting.

[Experience of using rhythmic transcranial magnetic stimulation, extracorporeal shock wave therapy and botulinotherapy in individual motor recovery programs in patients with spastic paresis of the lower limb]. / Opyt ispol'zovaniya v individual'nykh programmakh dvigatel'nogo vosstanovleniya ritmicheskoi transkranial'noi magnitnoi stimulyatsii, ekstrakorporal'noi udarno-volnovoi terapii i botulinoterapii u patsientov so spasticheskim parezom nizhnei konechnosti.

Walking disorder is one of the most frequent consequences of stroke and traumatic brain injury, occurring in 80% of cases. Spastic paresis of the muscles of the lower extremity is the cause formed in 20-40% of patients within a few weeks after brain damage. In this case, a complex of symptoms occurs: motor deficiency (muscle paresis), increased muscle tone (spasticity), biomechanical changes in muscles, joints and surrounding tissues, contractures. Recovery of walking is a difficult task due to the peculiarities of its organization in the norm. At the same time, changes occurring in the muscles of the lower limb after a stroke, their modular reorganization, the formation of various pathological patterns, violation of the regulation of movements by the central nervous system, rapidly occurring changes in muscles, ligaments, complicate this process. Improving walking is one of the most important priorities of rehabilitation. Already at the second (stationary) stage of rehabilitation, patients have a lack of proper support on the lower limb, which inevitably leads to excessive load on the second limb, a change in the body scheme, incorrect foot placement, violation of the mechanics of walking (moving from heel to toe) due to plantar flexion / turn of the foot, etc. All this makes patients dependent on outside help, and walking unsafe, increases the risk of falls and complications (arthropathy, contracture, etc.). In this regard, it is important to timely diagnose the totality of changes in the lower limb and create optimal comprehensive rehabilitation programs using highly effective treatment methods aimed at reducing the severity of the motor defect, reducing spasticity and preventing complications. The article discusses the place of rhythmic transcranial magnetic stimulation, extracorporeal shock wave therapy and botulinum therapy during rehabilitation in patients with spastic paresis of the lower limb after a stroke. The results of the protocol of clinical approbation «Complex rehabilitation of patients with lower limb spasticity after focal brain damage at the second stage of medical rehabilitation¼ are presented.