Pathological Changes in the Gastrocnemius Muscle throughout Hind Limb Ischemia Modeling in Rats.

We present a two-stage model for the study of chronic hind limb ischemia in rats. In the area of ischemia, sclerotic changes with atrophic rhabdomyocytes and reduced vascularization were revealed. CD31 expression in the endothelium increased proportionally to the number of vessels in the ischemic zone, and at the same time, focal expression of ßIII-tubulin was detected in the newly formed nerve fibers. These histological features are equivalent to the development of peripheral arterial disease in humans, which allows using our model in the search for new therapeutic strategies.

Aza-Heterocyclic Building Blocks with In-Ring CF2 -Fragment.

Modern organic chemistry is a titan supporting and reinforcing pharmaceutical, agricultural, food and material science products. Over the past decades, the organic compounds market has been evolving to meet all the research demands. In this regard, medicinal chemistry is especially dependent on available chemical space as subtle tuning of the molecule structure is required to create a drug with relevant physicochemical properties and a remarkable activity profile. The recent rapid evolution of synthetic methodology to deploy fluorine has brought fluorinated compounds to the spotlight of MedChem community. And now unique properties of fluorine still keep fascinating more and more as its justified installation into a molecular framework has a beneficial impact on membrane permeability, lipophilicity, metabolic stability, pharmacokinetic properties, conformation, pKa , etc. The backward influence of medicinal chemistry on organic synthesis has also changed the landscape of the latter towards new fluorinated topologies as well. Such complex relationships create a flexible and ever-changing ecosystem. Given that MedChem investigations strongly lean on the ability to reach suitable building blocks and the existence of reliable synthetic methods in this review we collected advances in the chemistry of respectful, but still enigmatic gem-difluorinated aza-heterocyclic building blocks.

Momentum Entanglement for Atom Interferometry.

Compared to light interferometers, the flux in cold-atom interferometers is low and the associated shot noise is large. Sensitivities beyond these limitations require the preparation of entangled atoms in different momentum modes. Here, we demonstrate a source of entangled atoms that is compatible with state-of-the-art interferometers. Entanglement is transferred from the spin degree of freedom of a Bose-Einstein condensate to well-separated momentum modes, witnessed by a squeezing parameter of -3.1(8) dB. Entanglement-enhanced atom interferometers promise unprecedented sensitivities for quantum gradiometers or gravitational wave detectors.

POSTOPERATIVE HEMORRHAGE AS A COMPLICATION OF A PARTIAL NEPHRECTOMY: FREQUENCY, FEATURES AND MANAGEMENT.

Objective - to study the character of possible postoperative complications and to define the reason and frequency of postoperative hemorrhage as a complication of partial nephrectomy. From January 2008 to December 2019 were performed 175 partial nephrectomy (PN) by a single surgeon in a high volume center. 41 operations were laparoscopic partial nephrectomy (LPN), 134 - open partial nephrectomy (OPN). In 152 cases kidney cancer was detected. Physical status, tumor volume, R.E.N.A.L. score, operative access, warm ischemia time (WIT), postoperative bleeding and its severity and treatment options were assessed in both groups of patients. Based on our study, R.E.N.A.L score may be a good tool in prognosis of a delay postoperative bleeding after nephron sparing surgery and this is statistically significant. On the other hand, single R.E.N.A.L score characteristics can't be reliable predictors of a delay bleeding. It is possible that a lack of cases with a significant postoperative bleeding in current study (6 of 175 cases) have some statistical restrictions. From our point of view, for better prognosis of delay bleeding, aside from hephrometric system it is important to take into account a proximity of a segmental arteries to a resection border, presens of any type of a coagulopathy and a preoperative antithrombotic therapy, obesity. High R.E.N.A.L score index is connected with a risk of significant postoperative bleeding, but this type of bleeding is rare after any nephron sparing surgery. Postoperative selective angioembolization is a method of choice and, in most cases, effective to stop kidney bleeding and nephron preservation.

The Cardio- and Neuroprotective Effects of Corvitin and 2-Oxoglutarate in Rats with Pituitrin-Isoproterenol-Induced Myocardial Damage.

Heart diseases, especially acute coronary syndrome (ACS), are among the most severe illnesses that often lead to death. Despite significant advances in the prevention and treatment of ACS, the incidence of the disease and its complications are very serious. The imbalance between pro- and antioxidant systems, the formation of active carbonyl compounds, and the end products of glycation in the blood and tissues are the key moments in the development of heart and neurological disorders leading to a change of behavioral responses. So, the search for antioxidants with cardio- and neuroprotective effects is an urgent task. This study was aimed at evaluating the effects of Corvitin and 2-oxoglutarate on physiological parameters, heart histology, and markers of carbonyl/oxidative stress of rats with pituitrin-isoproterenol-induced myocardial damage (PIMD). Increased sweating, tachycardia, significantly decreased locomotor and exploratory activity, changes of ECG, heart histology, and biochemical changes were observed in the PIMD-group. The administration of Corvitin or 2-OG led to the recovery of locomotor and cognitive activities of the rats, improvement in heart histology, a decrease in the levels of thiobarbituric acid reactive substances, advanced glycated end products, and various changes in the activity of the antioxidant enzymes, 6 days after PIMD. So, Corvitin and exogenous 2-OG show cardio- and neuroprotective effects through the decrease of carbonyl/oxidative stress and regulation of the activity of the antioxidant system.

[Differentiation of tortoises of the genera Testudo and Agrionemys (Testudinidae) based on the polymorphism of nuclear and mitochondrial markers].

Based on polymorphism of the 12S rRNA gene and RAPD markers, differentiation of 122 tortoise individuals belonging to the three species of genus Testudo (T. kleinmanni, T. marginata, and T. graeca), six subspecies of T. graeca (T. g. nikolskii, T. g. pallasi, T. g. armeniaca, T. g. zarudnyi, T. g. terrestris, T. g. ibera), and two subspecies of the Central Asian tortoise Agrionenemys horsfieldii (A. h. horsfieldii, A. h. kazakhstanica) was performed. For comparison, 32 known sequences of 12S rRNA gene (392 bp) from tortoises of the two genera inhabiting the territories of Europe, Asia, and Africa were used. In the populations of A. horsfieldii, a total of six haplotypes; including three newly described variants, were identified. In the examined tortoises of the genus Testudo, eleven 12S rRNA haplotypes were identified. One new haplotype was detected in T. kleinmanni. Among the eight subspecies of T. graeca, eight haplotypes were identified, with four newly described ones. The reported RAPD markers generally supported the reconstructions obtained with the use of the mitochondrial marker. Similarly to the 12S rRNA-based reconstructions, two independent clusters included representatives of the two genera, Agrionemys and Testudio. Among the latter, representatives of T. marginata and T. kleinmanni, as well as T. graeca, with high statistical support values, formed two reciprocally monophyletic groups. Compared to the mitochondrial markers, RAPDs more statisticallysignificantly discriminated the sample of T. g. terrestris and the four subspecies, T. g. ibera, T. g. armeniaca, T. g. pallasi, and T. g. nikolskii. In almost all cases except the representatives of T. g. ibera, the representatives of each of four subspecies formed individual subclusters. The geographical haplotype distribution patterns and possible evolutionary scenario of the origin and dispersal of tortoises of the two genera are discussed.

[Polymorphism of the 12S rRNA gene and phylogeography of the Central Asian tortoises Agrionemys horsfieldii gray, 1844].

Based on intraspecific polymorphism of 12S rRNA gene, genetic variation of isolated populations of the Central Asian tortoise, Agrionemys horsfieldii, was for the first time investigated on a large part of the species distribution range, encompassing Uzbekistan, southern Kazakhstan, and northern and eastern Iran. In 59 tortoises, four haplotypes were discovered, including two (AH1 and AH2), described earlier. Haplotype AH1 was detected in 52 tortoises, inhabiting southern Kazakhstan and Uzbekistan. Haplotype AH2 was found in four tortoises from the border territory between Uzbekistan, Tajikistan, and Afghanistan. Two novel haplotypes, AH3 and AH4, were detected in the three tortoises from Iran. Based on nucleotide substitutions in the 12S rDNA sequence, the possible divergence time between the tortoises from different parts of the range was estimated. Possible pathways of the formation of modern intraspecific groups of A. horsfieldii are discussed.

Topography of the active site of the Saccharomyces cerevisiae plasmalemmal dicarboxylate transporter studied using lipophilic derivatives of its substrates.

2-Alkylmalonates and O-acyl-L-malates have been found to competitively inhibit the dicarboxylate transporter of Saccharomyces cerevisiae cells, and the substrate derivatives chosen did not penetrate across the plasmalemma under the experiment conditions. Probing of the active site of this transporter has revealed a large lipophilic area stretching between the 0.72 to 2.5 nm from the substrate-binding site. Itaconate inhibited the transport fivefold more effectively than L-malate. This suggests the existence of a hydrophobic region immediately near the dicarboxylate-binding site (to 0.72 nm). The yeast plasmalemmal transporter was different from the rat liver mitochondrial dicarboxylate transporter. An area with variable lipophilicity adjoining the substrate-binding site has been revealed in the latter by a similar method. This area is mainly hydrophobic at distances up to 1.76 nm from the binding site and is separated by a hydrophilic region from 0.38 to 0.88 nm. Fumarate but not maleate competitively inhibited succinate transport into the S. cerevisiae cells. It is suggested that the plasmalemmal transporter binds the substrate in the trans-conformation. The prospects of the proposed approach for scanning lipophilic profiles of channels of different transporters are discussed.

Properties of yeast Saccharomyces cerevisiae plasma membrane dicarboxylate transporter.

Transport of succinate into Saccharomyces cerevisiae cells was determined using the endogenous coupled mitochondrial succinate oxidase system. The dependence of succinate oxidation rate on the substrate concentration was a curve with saturation. At neutral pH the K(m) value of the mitochondrial "succinate oxidase" was fivefold less than that of the cellular "succinate oxidase". O-Palmitoyl-L-malate, not penetrating across the plasma membrane, completely inhibited cell respiration in the presence of succinate but not glucose or pyruvate. The linear inhibition in Dixon plots indicates that the rate of succinate oxidation is limited by its transport across the plasmalemma. O-Palmitoyl-L-malate and L-malate were competitive inhibitors (the K(i) values were 6.6 +/- 1.3 microM and 17.5 +/- 1.1 mM, respectively). The rate of succinate transport was also competitively inhibited by the malonate derivative 2-undecyl malonate (K(i) = 7.8 +/- 1.2 microM) but not phosphate. Succinate transport across the plasma membrane of S. cerevisiae is not coupled with proton transport, but sodium ions are necessary. The plasma membrane of S. cerevisiae is established to have a carrier catalyzing the transport of dicarboxylates (succinate and possibly L-malate and malonate).

Study of active site topography of rat liver mitochondrial dicarboxylate transporter using lipophilic substrate derivatives.

Earlier it has been demonstrated that the active site (substrate-binding site + active site channel) of rat liver mitochondrial dicarboxylate transporter is characterized by rather complex topography. Probing the active site with 2-monoalkylmalonates revealed the existence of internal and external lipophilic areas separated by a polar region. A two substrate-binding site model of the transporter has been supposed. The correctness of this model has been evaluated by probing the active site with O-acyl-L-malates differing from 2-monoalkylmalonates by 0.23 nm longer distance from the anion groups to the aliphatic chain. Changes in the polar group of the probe did not prevent its binding and showed the same variable lipophilicity pattern for the transporter channel. Probing with alpha,omega-alkylene dimalonates did not reveal the second substrate-binding site at the active site. The substrate-binding site did not show any differences in affinity to O-acyl-derivatives of L-malate and D-malate, except L-malate binds more effectively than D-malate. This suggests involvement of the L-malate hydroxyl group in substrate binding and stereospecific behavior of the transporter substrate-binding site. A modified one substrate-binding site model of the dicarboxylate transporter is discussed.

Permeation of dicarboxylic acids with different terminal position of two carboxylic groups through planar bilayer lipid membranes.

Electrically silent hydrogen ion fluxes across a planar bilayer lipid membrane (BLM) induced by an addition of dicarboxylic (DC) acids at one side of BLM are monitored by measuring pH changes in the unstirred layers near the BLM surface via recording protonophore-dependent potentials. Two groups of DC acids are studied: (1) 2-n-alkylmalonic acids with an alkyl chain of different length which carry both carboxylic groups at one terminus of the hydrocarbon chain (alpha,alpha-DC acids); and (2) dicarboxylic acids of different linear chain length having carboxylic groups at the opposite ends of the hydrocarbon chain (alpha,omega-DC acids). It is shown that the pH optimum of hydrogen ion fluxes for the DC acids is shifted considerably to acidic pH values compared to monocarboxylic acids and is located near pH 5. For both types of DC acids at pH&z. Lt;5, the total transport is limited by diffusion of the anionic forms of the acids across the unstirred layers, while at pH&z.Gt;5 the transport is limited by diffusion of the neutral form across the membrane. The fluxes of alpha,alpha-DC acids are similar to those of alpha,omega-DC acids provided that the acids have the similar number of carbon atoms, the fluxes grow with the increase in the chain length of the alkyl radical.

The anion-carrier mediated uncoupling effect of dicarboxylic fatty acids in liver mitochondria depends on the position of the second carboxyl group.

Effects of dicarboxylic fatty acids with varying positions of the carboxyl groups on respiration and membrane potential of liver mitochondria were studied. Tetradecylmalonic acid (a fatty acid with two carboxyl groups in the alpha-position) efficiently uncoupled oxidative phosphorylation similarly to palmitic acid with the same number of carbon atoms. Similarly to the uncoupling by palmitic acid, the coupling effects of carboxyatractylate and glutamate changed reciprocally with changes in pH of the incubation medium: on increasing the pH from 7.0 to 7.8, the coupling effect of carboxyatractylate increased and that of glutamate decreased. A dicarboxylic fatty acid with the second carboxyl at the end of the alkyl chain in the omega-position (alpha, omega-tetradecyldicarboxylic acid) stimulated respiration of the mitochondria at a significantly higher concentration than myristic acid with the same number of carbon atoms, but unlike the latter nearly failed to decrease the transmembrane potential DeltaPsi. Neither carboxyatractylate nor glutamate inhibited the respiration stimulated by this dicarboxylic fatty acid.

[A method of determining the specific volume of liposomes with Turnbull blue]. / Metod opredeleniia udel'nogo ob''ema liposom s pomoshch'iu Turnbulevoisini.

A method for measuring the specific volume of liposomes by using ferricyanide as a maker is proposed. Ferricyanide is determined by the formation of Turnbull's blue. Advantages of the use of this reaction are an eightfold increase in the sensitivity and the possibility to determine the marker compound in the spectral region (730 nm) free of any considerable interference of natural compounds. Results of the use of this method for measuring the specific volume of liposomes prepared by ultrasonic treatment of a lipid suspension are reported.

[Uncoupling effect of dicarbonic analogs of stearic and palmitic acids in heart mitochondria]. / Razobshchaiushchee deistvie dikarbonovykh analogov stearinovoi i pal'mitinovoi kislot v mitokhondriiakh serdtsa.

The uncoupling effect of 2-hexadecylmalonic acid (HDMA) and 2-tetradecylmalonic acid (TDMA), the dicarboxylic derivatives of stearic and palmitic acids, have been studied in heart mitochondria. The effects of these compounds and related fatty acids on the induced by ascorbate oxidation with PMS (or TMPD) in the presence of oligomycin, phosphate and rotenone have been compared. It was found that both HDMA and TDMA uncoupled mitochondria with a similar efficiency as did the corresponding fatty acids, while the uncoupling effect of low concentrations of HDMA and TDMA were almost completely suppressed by carboxyatractylate.

Probing the active center of the mitochondrial dicarboxylate transporter.

2-n-Alkylmalonates with various length of the alkyl residue have been used to study the topography of the active center of the dicarboxylate transporter in intact rat liver mitochondria. Measurements of the Ki values of these competitive inhibitors suggest that in the transporter there is a large hydrophobic region at least 1.7 nm in size, containing a polar domain (ca. 0.5 nm) and situated close to a substrate-binding site. These zones are assumed to be involved in the mechanism of dicarboxylate transport.

[Characteristics of the interaction between 2-alkylmalonates and the substrate-binding site of the dicarboxylate carrier of rat liver mitochondria]. / Osobennosti vzaimodeistviia 2-alkilmalonatov s tsentrom sviazyvaniia substratov dikarboksilatnogo perenoschika mitokhondrii pecheni krys.

The effect of thirteen synthetic 2-n-alkyl derivatives of malonic acid on the rate of the mitochondrial succinate oxidase reaction controlled by the dicarboxylate carrier, was studied. These compounds were shown to act as competitive inhibitors of succinate transport and could interact with the carrier according to the 1:1 stoichiometry. The affinity of the inhibitor for the carrier increased in parallel with the increase in the alkyl residue length. This dependence was impaired only in the case of pentyl-, hexyl- and heptylmalonates having close values of inhibition constants. The data obtained suggest that the polar site and two hydrophobic regions are located in the vicinity of the carrier active site. The possible organization of the carrier substrate-binding site is discussed.