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.

[Study of stability of proline-containing derivatives of dopamine and serotonin in the biological media in vitro experiments]. / Issledovanie ustoichivosti prolinsoderzhashchikh proizvodnykh dofamina i serotonina v biologicheskikh sredakh v éksperimentakh in vitro.

Boc-Gly-Pro-DP, Z-Gly-Pro-DP, LA-Gly-Pro-DP, Boc-Gly-Pro-Srt, Z-Gly-Pro-Srt were synthesized for the first time. The stability of these compounds in the presence of leucine aminopeptidase, carboxypeptidase Y, carboxypeptidase B and proline endopeptidase (PEP) was determined. It turned out that the compounds are stable in the presence of aminopeptidases and carboxypeptidases. In the presence of PEP, dopamine (DP) and serotonin (Srt) are cleaved from the synthesized preparations. Thus, new proline-containing Srt and DP derivatives were obtained, Srt and DP could be gradually released from them. This suggest the possibility of a prolonged action of these biologically active compounds on the vital activity of cells and, consequently, of the whole organism.

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.

[Stability of prolin-containing peptides in biological media]. / Ustoichivost' prolinsoderzhashchikh peptidov v biologicheskikh sredakh.

New data on peptide drugs have been summarized; their high stability is due to both the introduction of Pro-Gly-Pro in various amino acid sequences and the modification of the glyproline fragment itself. Pro-Gly-Pro-Leu, ACTH(6-9)Pro-Gly-Pro, 5-oxo-Pro-Arg-Pro and 5-oxo-Pro-His-Pro-NH2 were used as proline-containing peptides. Tritiated peptides were obtained: Pro-Gly-Pro-Leu with specific radioactivity of 135 Ci/mmol, ACTH(6-9)Pro-Gly-Pro - 26 Ci/mmol, 5-oxo-Pro-Arg-Pro - 60 Ci/mmol and 5-oxo-Pro-His-Pro-NH2 - 75 Ci/mmol. The concentration of Pro-Gly-Pro-Leu, ACTH(6-9)Pro-Gly-Pro, 5-oxo-Pro-Arg-Pro and 5-oxo-Pro-His-Pro-NH2 in the blood was found to be about 200 times more than in the brain for intranasal administration, and in average 600 times more for intravenous administration. The stability of proline-containing peptides in vitro experiments was determined using different commercially available peptidases (leucine aminopeptidases, dipeptidases, carboxypeptidases B and Y), and using nasal mucus, microsomal fraction of the rat brain (IMPC) and rat blood plasma. During peptidase hydrolysis of Pro-Gly-Pro-Leu, the main metabolites were Gly-Pro-Leu, Pro-Gly-Pro, Gly-Pro and Pro-Gly. For ACTH(6-9)Pro-Gly-Pro, the main metabolites were Phe-Arg-Trp-Pro-Gly-Pro and Trp-Pro-Gly-Pro. In peptidase hydrolysis of 5-oxo-Pro-His-Pro-NH2, the major metabolite was 5-oxo-Pro-His-Pro. It was shown that with different methods of peptides administration the composition of the metabolites formed is different. Based on the data obtained, resistance to enzymatic cleavage of peptides and their metabolic pathways were evaluated. Thus, these new data have shown that the above approaches can be used to prolong the action of glyprolines in living objects. In this case, the degradation of proline-containing peptides occurs mainly not due to the action of proteases, but due to other ways of degradation. In general, the data presented in the review indicate the promise of intranasal way of introducing biologically active peptides into the brain of living organisms.

[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.

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.

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.

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.

[BLOCKING SENSORIMOTOR DISORDERS IN STRIATIC DOPAMINERGIC DEFICIENT MICE BY THE EXTRACTS OF "REMEDY BEETLE" ALPHITOBIUS DIAPERINUS].

Five groups of C57Bl/6JSto mice (groups 1-5) were tested on a vertical pole 2 weeks since subcutaneous administration of proneurotoxin MPTP (40 mg per kg b. w.) to groups 2-5. Group 1 served as control. It is known that this MPTP dosage forms within 2 weeks sensorimotor disturbances similar to the initial stage of Parkinson's disease. One week before MPTP injection and during all the period after it (2 weeks) animals of groups 3-5 got one of the three extracts of biomass homogenate of the «remedy beetle¼ Alphitobius diaperinus as an additive to their food (8 g per 1 kg of food). The extracts were as follows: water (N 1), water-methanol (N 2) and water-methanol one after solid-phase extraction (N 3). All extracts were immobilized on a phytocarrier. Animals of groups 1 & 2 used ordinary food. Sensorimotor disturbances which could be clearly seen in the 2nd group of mice (MPTP) were absent in group 1 (control) as well as groups 3-5 which received an «antidote¼ with their food as one of the extracts of homogenate of the beetle biomass. The extract N 3 demonstrated maximal efficiency of all.

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.

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.

[Criteria of antibacterials choice for prophylaxis and therapy of implant-associated infections in prosthetic joints].

Prosthetic joints infection is a serious medical, social and financial problem. Significant achievements in prophylaxis of such infections are well known. In cases of primary endoprosthetics of large joints, the frequency of infection in the leading world clinics is 0.5-2%. However, the approaches to the treatment of such infections are different especially with respect to the antibacterial therapy: choice of drugs, administration routes, treatment course and discontinuation. Under conditions of global growth of resistance in nosocomial pathogens, the choice of antimicrobials requires consideration not only of the formal susceptibility criteria, but also of the pharmacokinetic and pharmacodynamic indices.

Interaction of holothurian triterpene glycoside with biomembranes of mouse immune cells.

The in vitro interactions between triterpene glycoside, cucumarioside A(2)-2, isolated from the Far-Eastern holothurian Cucumaria japonica, and mouse splenocyte and peritoneal macrophage biomembranes were studied. Multiple experimental approaches were employed, including determination of biomembrane microviscosity, membrane potential and Ca(2+) signaling, and radioligand binding assays. Cucumarioside A(2)-2 exhibited strong cytotoxic effect in the micromolar range of concentrations and showed pronounced immunomodulatory activity in the nanomolar concentration range. It was established that the cucumarioside A(2)-2 effectively interacted with immune cells and increased the cellular biomembrane microviscosity. This interaction led to a dose-dependent reversible shift in cellular membrane potential and temporary biomembrane depolarization; and an increase in [Ca(2+)](i) in the cytoplasm. It is suggested that there are at least two binding sites for [(3)H]-cucumarioside A(2)-2 on cellular membranes corresponding to different biomembrane components: a low affinity site match to membrane cholesterol that is responsible for the cytotoxic properties, and a high affinity site corresponding to a hypothetical receptor that is responsible for immunostimulation.

[The role of splenectomy in the treatment of idiopatic thrombocytopenic purpura].

Based on the earlier published data this article describes the role of the spleen in the development of some hematological diseases, the main of which is idiopathic thrombocytopenic purpura (ITP). The authors emphasize the role of splenectomy in the treatment of this disease. At 178 splenectomies performed at the clinic, 23,6 % of operations were carried out for hematological diseases, and among them 42,8 %--for ITP. The authors share their immediate and remote results of the treatment.