Evaluation of the five-year Bloomberg Philanthropies Global Road Safety Program in the Russian Federation.

Road traffic injuries are a leading cause of morbidity and mortality in the world. In Russia, a road safety program was implemented in Lipetskaya and Ivanovskaya oblasts (regions) as part of a 10-country effort funded by Bloomberg Philanthropies. The program was focused on increasing seat belt and child restraint use and reducing speeding. The primary goals of this monitoring and evaluation study are to assess trends in seat belt use, child restraint use, and speed compliance in the two oblasts over the 5 years and to explore the overall impact of the program on road traffic injury and death rates. Primary data via roadside observations and interviews, and secondary data from official government sources were collected and analyzed for this study. Our results indicate significant improvements in seat belt wearing and child seat use rates and in prevalence of speeding in both intervention oblasts. The observations were consistent with the results from the roadside interviews. In Lipetskaya, restraint use by all occupants increased from 52.4% (baseline, October 2010) to 77.4% (final round, October 2014) and child restraint use increased from 20.9% to 54.1% during the same period. In Ivanovskaya, restraint use by all occupants increased from 48% (baseline, April 2012) to 88.7% (final round, October 2014) and child restraint use increased from 20.6% to 89.4% during the same period. In Lipetskaya, the overall prevalence of speeding (vehicles driving above speed limit) declined from 47.0% (baseline, July 2011) to 30.4% (final round, October 2014) and a similar pattern was observed in Ivanovskaya where the prevalence of speeding decreased from 54.6% (baseline, March 2012) to 46.6% (final round, October 2014). Through 2010-2014, the road traffic crash and injury rates per 100,000 population decreased in Lipetskaya oblast (191.5 and 246.9 in 2010 and 170.4 and 208.6 in 2014, respectively) and slightly increased in Ivanovskaya oblast (184.4 and 236.0 in 2010 and 186.7 and 243.4 in 2014, respectively). These road safety improvements are associated with the program that enabled a combined focus on policy reform, legislation, enforcement, advocacy, education, and data collection and use. However, the existence of other road safety efforts, lack of data from comparable regions, and unavailability of risk factor-specific data make it difficult to attribute these changes to the program.

[Isolation and purification of human blood plasma proteins able to form potassium channels in artificial bilayer lipid membrane].

Protein fraction able to induce K(+)-selective transport across bilayer lipid membrane was isolated from human blood plasma with the use of the detergent and proteolytic enzyme-free method developed at our laboratory. After addition of the studied sample to the artificial membrane in the presence of 100 mM KCl, a discrete current change was observed. No channel activity was recorded in the presence of calcium and sodium ions. Channel forming activity of fraction was observed only in the presence of K+. Using a threefold gradient of KCl in the presence of studied proteins the potassium-selective potential balanced by voltage of -29 mV was registered. This value is very close to the theoretical Nernst potential in this case. This means that the examined ion channel is cation-selective. According to data obtained with MS-MALDI-TOF/TOF and database NCBI three protein components were identified in isolated researched sample.

Activity of mitochondrial ATP-dependent potassium channel in animals with different resistance to hypoxia before and after the course of hypoxic training.

Activity of mitochondrial ATP-dependent potassium channel in rats with high genetically determined resistance to hypoxia was higher than in sensitive animals. Adaptation of low resistant rats to hypoxia was accompanied by activation of the channel, facilitation of potassium recycling in mitochondria, and a decrease in the rate of H2O2 formation. Our results indicate that mitochondrial ATP-dependent potassium channel plays an important role in the delayed mechanisms of animal's adaptation to hypoxia.

[Localization in a cell of a protein forming the ATP-dependent potassium-selective channels in the bilayer lipid membrane. An ultrastructural study].

The localization in the cell of the protein forming the ATP-dependent potassium-selective channels in the bilayer lipid membrane has been studied. The electron microscope investigation of rat liver and heart tissue sections after their incubation with Abs against this protein and the visualization of the protein with secondary Abs conjugated with colloid gold were carried out. Colloid gold particles were observed both in mitochondrial membranes and in membranes of endoplasmic and sarcoplasmic reticulum. In heart mitochondria, these particles were significantly greater than in liver mitochondria. The localization of the channel protein both in mitochondria and reticulum, as well as the structural similarity between the mitochondrial channel and the precursor of calreticulin suggest that the channel protein belongs to the family of calreticulins. The possible function of the protein as a channel subunit of the mitochondrial ATP-dependent potassium channel is discussed.

[Purification of the mitochondrial calcium uniporter from the beef heart and characterization of its properties].

A low-molecular-weight component (LMC) inducing selective transport of calcium across the bilayer lipid membrane has been isolated from mitochondria of the bovine heart by the method developed in our laboratory, which excludes the use of detergents and proteolytic enzymes. It was shown that, in the presence of 10 mM CaCl2, LMC forms conduction channels in the membrane multiples of 5 pS. The specific inhibitor of mitochondrial calcium uniporter, ruthenium red, closes Ca2(+)-induced channels formed in the membrane by LMC. In the absence of calcium or in the presence of potassium ions only, the component is incapable of forming channels of conduction. It was shown using nuclear magnetic resonance that LMC is a complex consisting of lipids, amino acids, and sugars with a molecular weight of 1-2 kDa.

[Effect of hypoxenum on bioenergetic processes in mitochondria and the activity of ATP-sensitive potassium channel].

The effect of hypoxenum on bioenergetic processes in heart and liver mitochondria of rats, connected with respiration, the generation of hydrogen peroxide, and the activity of ATP-sensitive K-channel ((mitoK)ATP) has been studied. It was shown that hypoxenum in the concentration range of 0.05-10 microg/ml stimulates respiration, increases the coupling in the respiratory chain, and enhances the formation of H2O2 and energy-dependent swelling associated with potassium transport in mitochondria. Hypoxenum removes the inhibitory effect of ATP on the energy-dependent swelling of mitochondria and partially reduces the accumulation of H2O2 in the presence of ATP. The role of antihypoxic and antioxidant action of hypoxenum associated with the activation of (mitoK)ATP is discussed.

[Age-related changes in the functioning of the mitochondrial potassium-transporting system].

Changes in the rate of respiration and functioning of the ATP-dependent potassium channel the liver and heart mitochondria of one-, three-, eight-, and twenty four-month-old Wistar male rats have been investigated. It was shown that the activity of the channel in the mitochondria of both tissues in 24-months-old animals decreases more than three times, and the content of potassium, 1.5-2 times compared with young one-month-old rats. The changes occur against the background of age-related decrease of energy supply in mitochondria, the respiratory complex-I underganig the greatest changes upon aging. The decrease of channel activity may be the result of changes in channel sensitivity to modulators and a decrease in the expression of mitochondrial K(+)-transporting channel-protein with a molecular mass of 5.5 kDa upon aging found in this work. As a result, the functioning of not only the mitoK(ATP) but also the whole mitochondrial potassium cycle is impaired.

Effect of several flavonoid-containing plant preparations on activity of mitochondrial ATP-dependent potassium channel.

Flavonoid-containing plant preparations (water soluble extracts of Pentaphylloides fruticosa [Extralife], Emblica officinalis Gaerth [Amla], and Bergenia crassifolia [Bergenia]) produced a dose-dependent and tissue-specific effect on activity of mitochondrial ATP-dependent potassium channel. The effect of these preparations was biphasic (activation and inhibition). The activating effect of Extralife was one order of magnitude higher than that of Amla and Bergenia and was observed in a wider concentration range. The activating effect of preparations was abolished by inhibitors of the mitochondrial ATP-dependent potassium channel, which attested to specificity of their influence on mitochondrial channel. Under in vivo conditions, the antihypoxic effect of Extralife was partially abolished by mitochondrial ATP-dependent potassium channel inhibitor 5-hydroxydecanoate.

[Changes in macro- and micro element composition in the organs of irradiated animals]. / Izmenenie makro- i mikroélementnogo sostava v organakh obluchennykh zhivotnykh.

We studied the content of Al, Cd, Co, Cu, Fe, Mg, Mn, P, Pb, S, and Zn in the liver, kidneys, lungs, and spleen of mice with total radiation dose of 7.5 Gy using atomic emission spectral analysis with an inductively coupled argon plasma. The qualitative content of macro- and microelements and coordination between their concentrations statistically determined from coefficients of linear correlation differ between the tissues of the irradiated and control animals. Radiation damage decreases phosphorus content in all studied organisms and is a marker of disturbed conjugated oxidative phosphorylation. The most significant radiation-induced disturbance of macro- and microelement balance was detected in the spleen; it features decreased content of phosphorus, magnesium, and cobalt, as well as increased content of zinc and aluminum and considerably increased iron content. The revealed macro- and microelements disbalance in spleen, liver, kidneys, and lungs can be considered as a test for primary biological response to radiation damage.

Palmitic and stearic acids bind Ca2+ with high affinity and form nonspecific channels in black-lipid membranes. Possible relation to Ca2+-activated mitochondrial pores.

A mitochondrial hydrophobic component that forms Ca2+-induced nonspecific ion channels in black-lipid membranes (Mironova et al., 1997) has been purified and its nature elucidated. It consists of long-chain saturated fatty acids--mainly palmitic and stearic. These fatty acids, similar to the mitochondrial hydrophobic component, bind Ca2+ with high affinity in comparison with unsaturated fatty acids, saturated fatty acids with shorter aliphatic chains, phospholipids, and other lipids. Ca2+-binding is inhibited by Mg2+ but not by K+. For palmitic acid, the Kd for Ca2+ was 5 microM at pH 8.5 and 15 microM at pH 7.5, with the Bmax of 0.48 +/- 0.08 mmol/g. This corresponds to one Ca2+ ion for eight palmitic acid molecules. The data of IR spectroscopy confirm that Ca2+ does not form ionic bonds with palmitic and stearic acids under hydrophobic conditions. It has been found that in the presence of Ca2+, palmitic and stearic acids, but not unsaturated FFA induce a nonspecific permeability in black-lipid membranes. Addition of Ca2+ in order to induce the permeability transition, increases the extractable amount of palmitic and stearic acids, the effect being prevented by a phospholipase A2 inhibitor. The possible involvement of palmitic and stearic acids in the mitochondrial nonspecific permeability is discussed.

[On vitamin B1 metabolism in avitaminosis and its correction with thiamine and taurine]. / K voprosu metabolizma vitamina B1 v usloviiakh avitaminoza i ego korrektsii tiaminom i taurinom.

The levels of phosphate esters and the activities of thiamine biotransformation enzymes in the blood and tissues of albino rats were studied during oxythiamine-induced B1 deficiency and after metabolic correction with thiamine and taurine. Among thiamine phosphates, the most informative indicators of thiamine deficiency were shown to be triphosphate esters and free thiamine diphosphate. The biosynthetic enzymes thiamine kinase and thiamine diphosphate kinase played a decisive role in maintaining the initial rate and in recovering the physiologically active forms of vitamin B1. The activation of hydrolytic enzymes of thiamine phosphate esters occurred by producing abundant free thiamine diphosphate and thiamine triphosphate. Within the first hours, taurine favoured the acceleration of phosphoester biosynthesis and, accumulating in the tissues, inhibited vitamin phosphorylation reactions.

[Metabolism of vitamin B1 and its di- and triphosphate esters in experimental allergic encephalomyelitis]. / Metabolizm vitamina B1 i ego di- i trifosfornogo efirov pri éksperimental'nom allergicheskom éntsefalomielite.

Biological significance of thiamin in development of experimental allergic encephalomyelitis has been elucidated. It has been shown that at the late preclinical stage of the disease thiamine metabolism is predominantly directed towards the maintaining of cellular metabolic homeostasis, whereas at the stage of clinical symptoms the anabolic process gives way to catabolic decomposition. Among tested thiamine phosphates the triphosphate ester is the most informative parameter in demyelinizing processes. Thiamine injections to immunized animals accelerate the vitamin phosphorylation depleting the reducing and energy potentials of the cell. Such thiamine antagonist as oxythiamine inhibits phosphatase reactions.

[Metabolism of vitamins B1 and PP and their use in oncological practice]. / Obmen vitaminov B1 i PP i ikh primenenie v onkologicheskoi praktike.

Blood of patients with gastric tumor was studied after their admission to the hospital and after the chemotherapeutic course. Formation of the tumor was accompanied by development of hypovitaminoses B1 and PP. The vitamin deficiency was more distinct after treatment of the patients with cyclophosphan: content of thiamine diphosphate (TDP) was decreased by 40%; NAD+NADP, by 30% and NADH+NADPH, by 20%. In mice with Ehrlich ascites carcinoma, activity of transketolase in erythrocytes was decreased by 48%, content of TDP, by 61% and that of NADPH, by 27%. The administration of cyclophosphan increased further thiamine deficiency in the tumor-bearing mice. Simultaneous administration of thiamine and cyclophosphan abolished the cytostatic toxic effect but did not affect their antitumoral properties. Under these conditions treatment with vitamins B1 and PP complex was undesirable due to malignization. The vitamins B1 and PP did not stimulate the tumor growth, partially restored impaired metabolism of the vitamins and may be included separately into combined multidrug oncotherapeutics.

[Fermentation micromethod for the quantitative determination of thiamine diphosphate in biological fluids]. / Fermentativnyi mikrometod kolichestvennogo opredeleniia tiamindifosfata v viologicheskikh zhidkostiakh.

An enzymatic micromethod is proposed for quantification of thiamine biphosphate (TBP) at concentrations from 0.5 ng in 0.1-0.2 ml samples of blood or other biological liquids. The dynamics of TBP degradation in blood was studied depending on the time and conditions of storage. A high efficient complex of alcohol dehydrogenase and apopyruvate decarboxylase was isolated from baker's yeasts that can be successfully used for quantitative detection of TBP. The complex was stabilized for further application to biochemical kits for diagnosis of B1-deficiency.

[Metabolism of thiamine and its di- and triphosphorus esters in the blood of patients with neurological symptoms of osteochondrosis]. / Metabolizm tiamina, ego di- i tri-fosfornogo éfirov v krovi bol'nykh s nevrologicheskimi proiavleniiami osteokhondroza.

Concentrations of phosphate esters in blood of patients with vertebral osteochondrosis and activities of enzymes involved in thiamine metabolism have been investigated. The pathological processes were characterized by a decrease in content of thiamine triphosphate while concentration of coenzyme, thiamine diphosphate, was relatively constant. The enzymes involved in the initial and final steps of biosynthesis and degradation of thiamine phosphorylated derivatives play the main role in maintaining vitamin B1 metabolic homeostasis.

[Several aspects of early ventricular repolarization syndrome]. / Nekotorye aspekty sindroma rannei repoliarizatsii zheludochkov.

An epidemiological study was undertaken to examine the prevalence of the early ventricular repolarization syndrome (EVRS) among 3,105 subjects. An analysis was made of 796 ECG showing EVRS. The genesis of EVRS was interpreted in terms of vector-scalar and chronotopographic analyses of myocardial de- and repolarization. The syndrome was also characterized from clinical and electrophysiological points of view.

[Thiamine triphosphate content of the liver of albino rats with different levels of thiamine]. / Soderzhanie tiamintrifosfata v pecheni belykh krys pri pazlichnoi obespechennosti tiaminom.

Content of thiamine triphosphate (TTP) was studied in rat liver tissue under conditions of normal state, thiamine deficiency and after loading with thiamine. The concentration of TTP in hepatocytes of control animals was found to be 3.2-3.6 micrograms/g, corresponding to 40% of the total thiamine pool in the cells. The TTP appears to serve as a reserve of vitamin B1 deposited in mitochondria of the cells in which the biosynthesis of thiamine pyrophosphate of their own did not take place. Administration of thiamine did not induce the TTP overproduction in hepatocytes; the content of TTP was maintained in the mitochondria of the hepatocytes at the constant level.