Modern Approaches for Identification of Modified Nucleotides in RNA.

This review considers approaches for detection of modified monomers in the RNA structure of living organisms. Recently, some data on dynamic alterations in the pool of modifications of the key RNA species that depend on external factors affecting the cells and physiological conditions of the whole organism have been accumulated. The recent studies have presented experimental data on relationship between the mechanisms of formation of modified/minor nucleotides of RNA in mammalian cells and the development of various pathologies. The development of novel methods for detection of chemical modifications of RNA nucleotides in the cells of living organisms and accumulation of knowledge on the contribution of modified monomers to metabolism and functioning of individual RNA species establish the basis for creation of novel diagnostic and therapeutic approaches. This review includes a short description of routine methods for determination of modified nucleotides in RNA and considers in detail modern approaches that enable not only detection but also quantitative assessment of the modification level of various nucleotides in individual RNA species.

[Immunogenicity of recombinant analog of antitumor protein lactaptin].

Therapeutic monoclonal antibodies and recombinant proteins including cytokines are commonly used in the treatment of cancer and inflammatory diseases. In most cases, these protein-based drugs exhibit a high therapeutic efficacy, which is unfortunately frequently associated with a variety of side effects. We have investigated the in vitro and in vivo immunogenicity of recombinant antitumor protein lactaptin (RL2). Based on the qRT-PCR analysis, we have shown that, in MDA-MB-231 human breast adenocarcinoma cells, RL2 suppresses the NF-kB signaling cascade that regulates the reactions of innate immunity. RL2 inhibits the expression of the CXCL1 protein and apoptosis inhibitor A20 and enhances expression of IkB, NF-kB repressor. The ELISA method has been used to evaluate the antibody titer in the blood of mice, which received single and triple intravenous or intraperitoneal injections of RL2. The multiplex immunoassay of 23 cytokines in the mice blood has shown that the RL2 injections lead to a slight increase in the levels of systemic pro-inflammatory cytokine interleukin-5 (IL-5) and keratinocyte chemoattractant (KC), a homologue of human macrophage inflammatory protein-1 (MIP-1). These observations indicate the low immunogenicity of the recombinant lactaptin analog, which can be considered to be a potential molecular drug candidate for further clinical development.

[Laparoscopic transperitoneal adrenalectomy].

AIM: To analyze the results of laparoscopic adrenalectomy (LAE) in patients with adrenal gland tumors. MATERIALS AND METHODS: From 2011 to 2016, 24 patients (15 men, 9 women), mean age 49.6 +/- 8.2 years, underwent LAE. The right, left and bilateral LAE was performed in 12, 10 and 2 patients, respectively. Indications for surgery were mainly primary and metastatic adrenal tumors. The operations were performed using a transperitoneal 4-port approach. RESULTS: Twenty six LAE were successfully performed in 24 patients. There were no conversions. Blood transfusion was used only in 1 patient, who had a history of the laparoscopic left nephrectomy for the renal cell carcinoma 2 years earlier. She had bleeding due to a spleen injury during the left LAE. She also developed acute postoperative pancreatitis, resolved by conservative therapy. The mean intra- and postoperative blood loss was 160 (120-750) ml, the operative time was 135 (100-205) min, and the length of hospital stay was 4 (3-5) days. Histological examination revealed adenocarcinoma and adenoma in 20 (73.1%) and 6 (26.9%) removed adrenal glands, respectively. CONCLUSION: Laparoscopic adrenalectomy is an effective and safe surgical modality to treat patients with adrenal tumors.

[The influence of recombinant nucleophosmin 1 on artificial RNA internalization into human adenocarcinoma MCF-7 cells].

In this study we obtained and characterized the recombinant analogue of multifunctional nucleolar phosphoprotein nucleophosmin 1 (NPM1) involved in crucial cellular processes such as transcription, reparation and mitosis. The influence ofnucleophosmin 1 on extrcellular RNAs accumulation in human adenocarcinoma cells MCF-7 was analyzed. It was found that incubation of AluY RNA (n > 300 nt), U24 snoRNA analogues (n ~ 80 nt) with Npm1-His6 resulted in RNA-protein non-covalent complexes formation, but not in case of the short oligoribonucleotide (n = 22 nt). It was shown that interaction of AluY RNA analogue with Npm1-His6 significantly increases transfection efficacy of the RNA into MCF-7 human cells. Altogether, these data allow us to conclude, that nucleophosmin 1 not only binds RNA with complex secondary structure, but also promotes uptake and internalization of such RNA by human cells.

[Renal revascularization and long-term cardiovascular survival of patients with atherosclerotic renovascular disease].

The prospective investigation of 185 patients (118 men and 67 women in the age of 35-79 years (on the average 60+/-9 years) with atherosclerotic renovascular disease was carried out. The disease was detected by one- (62%) and two- (38%) hemodynamic value (>50%) of renal artery stenosis. Renal function, arterial pressure, cardiovascular death cases were estimated in 4 groups of patients during the period of 2,5-126 months (on the average 33 months). The patients of the first group (n=103) were undergone the renal revascularization (51 open and 80 endovascular). The patients of the second group (n=20) were after combined open renal revascularization (25) and after performed reconstruction of abdominal aorta and its branches and lower extremities arteries concerning aneurysmatic and occlusive lesions. The patients of the third group (n=19) had only reconstruction of lower extremities arteries concerning their ischemia without renal revascularization. The patients of the fourth group (n=43) haven't been operated because of the refusal of intervention. The lowest rate of survival was in the fourth group, who used the conservative therapy. Long-term survival didn't differ after open and endovascular therapy. The Koks regression analysis showed, that the improvement of long-term survival of the patients was associated with renal revascularization [Exp(B)=0,25, p<0,001] and didn't depend on the method of renal arteries reconstruction. A strategy of careful selection of patients with the application of interdisciplinary approach should be the base of clinical practice in renovascular disease.

[The analysis of biochemical markers of MCF-7 cells apoptosis induced by recombinant analog of lactaptin].

Earlier we isolated and characterized human milk pro-apoptotic peptide - lactaptin and generated its engineered analog - RL2. It was shown that both lactaptin and RL2 are capable to induce apoptotic death of MCF-7 cells. In this report we have analyzed biochemical markers of RL2 induced MCF-7 apoptosis. The activation of initiator and effector caspases as well as mitochondrial membrane potential and cytoplasm membrane changes were analyzed using flow cytometry and Western-blot methods. We have found that RL2 induced apoptotic death of MCF-7 cells was accompanied by PS exposure on the plasma membrane surface. It also was shown that RL2 has induced dissipation of mitochondrial membrane potential and resulted in activation of initiator caspases 8, 9 and effector caspase 7.

[Results of combined reconstructions of the renal arteries, abdominal aorta and its other branches in patients with atherosclerosis].

A comparative analysis was made of clinical indices and long-term survival in 20 patients (13 men and 7 women of mean age 62 +/- 2 years), having multifocal atherosclerosis of the renal arteries (RA), abdominal aorta and arteries of lower extremities (LE) (11 aneurysms, 7 occlusive lesions of the aorta and 3 of its unpaired branches) who underwent combined reconstructions, with patients having similar lesions, but not operated (12) or operated only on RA or on the arteries of LE (45). The groups of patients had no great differences except more pronounced renal dysfunction (RD) in the first of them. In spite of high postoperative lethality (20%) mainly due to acute renal failure (3), long-term survival proved to be significantly higher (14 from 16), than in not operated (3 from 12) (p = 0.00001) and had better tendency than in those operated on the same basin (35 from 45). In addition, the patients after combined operations had substantially decreased systolic and diastolic arterial pressure (159 +/- 13 vs. 132 +/- 9, p = 0.00001; 91 +/- 5 vs. 83 +/- 7, p = 0.010) and also stabilized renal function.

[The effect of the use of the drug recognan (citicoline) on the state of higher mental functions in patients with mild cognitive impairment]. / Vliyanie preparata Rekognan na sostoyanie vysshikh psikhicheskikh funktsii u patsientov s legkimi kognitivnymi narusheniyami.

OBJECTIVE: The aim of the observational program was to study the effect of the use of the drug recognan (citicoline) on the state of higher mental functions (memory, attention, visual-motor coordination, dynamic praxis,verbal thinking and imagination) in patients with mild cognitive impairment. MATERIAL AND METHODS: A survey of 54 subjects (16 of them male and 38 female) aged 18-50 years (average age 28.5±10.5 years) was conducted with a diagnosis of «Mild cognitive impairment¼ («F06.7¼). The group was randomized into 2 subgroups: the main subgroup (26 people) received oral therapy with the drug Recognan, for 30 days, with the daily dosage of the drug being 500 mg. In the control group (28 people), nootropic drug therapy was not performed. Standard psychometric techniques were used to study higher mental functions. All subjects were examined three times (initially, in the middle of the study - on day 15, at the end of the study - on day 30). RESULTS AND CONCLUSION: After 2 weeks of treatment with recognan, there was an improvement in concentration in 81.9%, memory in 50% (p=0.008), verbal imagination productivity in 68.2% (p=0.015), counting functions in 60% (p=0.015), visual-motor coordination and dynamic praxis - in 86.4% (p=0.003), increased speed and efficiency of mental work (p=0.001). After a 30-day course of treatment with recognan, there was an improvement in memory in 58.3% of patients (p=0.007), an increase in concentration in 64%, an improvement in counting functions in 64.3% (p=0.011), verbal imagination productivity in 63.3%, visual-motor coordination and dynamic praxis in 86.4% (p=0.007), speed and efficiency of mental work (p=0.006), which indicates a complex positive effect of recognan on higher mental functions in patients with mild cognitive impairment cognitive impairment.

[The influence of the drug recognan (citicoline) on neurodynamic characteristics of mental activity in patients with mild cognitive impairment]. / Vliyanie preparata Rekognan (tsitikolin) na neirodinamicheskie kharakteristiki psikhicheskoi deyatel'nosti u patsientov s legkimi kognitivnymi narusheniyami.

OBJECTIVE: The aim of the study was to study the effect of the drug recognan (citicoline) on the neurodynamic characteristics of mental activity in patients with mild cognitive impairment. MATERIAL AND METHODS: A survey of 58 subjects (17 of them male and 41 female) aged 18-45 years (average age 27.2±12.5 years) was conducted. Clinical diagnosis according to ICD-10 «Mild cognitive impairment¼ (F06.7). The main subgroup included29 people received oral recognan therapy (in solution, 100 mg in 1 ml) for 30 days, with a daily dosage of 500 mg (5 ml of solution). In the control group (29 people) drug therapy was not performed. Tests were used: «Graphic sample¼, the sample on the reciprocal coordination of the hands (Ozeretsky test), test for the compression of fingers, the test «number series¼. The follow-up period was 30 days. All subjects were examined three times (initially, in the middle of the study - on day 15, at the end of the study - on day 30). RESULTS AND CONCLUSION: The results obtained showed that the use of the drug recognan (citicoline) has a positive effect on the indicators of visual-motor coordination and spatial representations, neurodynamic characteristics of movement, and cognitive functions. After a 2-week treatment with recognan improved graphical sample, 84% of patients, with reliable improvement according to the Wilcoxon (p=0.0002), the sample in the compression of the fingers 60%, as well as coordination 60%, the accounting functions in 44%. After a month (30 day) treatment course recomanem there was an increase in indices of samples for the compression of fingers in 71.4% of patients, with significant improvement (p=0.0499) and run the graphical samples of 71.4%, coordination at 46.4%, counting functions - 48% patients.

[Novel method for activation of the locomotor circuitry in human].

We examine the possibility for activation of the involuntary locomotion of the lower limbs by spinal electromagnetic stimulation (ES). The subject laid on the left side. The legs are supported in a gravity-neutral position by special mounting that to provide horizontal rotation in the hip, knee and ankle. ES (3 Hz and 1.56 Tesla) at the T11,-T12 vertebrae induced involuntary locomotor-like movements in the legs. The latency from the initiation of ES to the first EMG burst compoused 0.68 +/- 1.0 s and it shortened at increasing of the frequency ES from 3 Hz to 20 Hz. Thus, the spinal ES can unduce the activation of the locomotor movements in human.

[Rationale for using essential phospholipids in chronic diseases of the liver: the dynamics of electric and viscoelastic parameters of erythrocytes].

Under supervision there were 38 men (age from 36 till 57 years) with diffuse hepatic diseases (DHD) in stadia of hepatitis, 27--in dynamics of treatment of essential phospholipids. Is experimentally established: structure-functional erythrocyte characteristics by methods of dielectrophoresis, thin-layer chromatography correlate with lipid and phospholipid composition of red cell membranes. The low level of phospholipids in erythrocyte membranes and direction of changes in their fractional composition in the DHD is the basis for the use of essential phospholipids. Against the background of therapy noted a significant increase in amplitude of deformation, polarizability, capacity, speed of movement of red blood cells to the electrodes and lower levels of generalized indicators of viscosity, rigidity, electrical conductivity, index of aggregation and destruction.

[Surgical treatment of fibromuscular dysplasia of renal arteries in adults].

The authors have analyzed an experience with treatment of 16 patients. All the patients had renovascular hypertension (RVH) and 3 of them had renal dysfunction. Revascularization of the kidneys (RK) was fulfilled in 11 patients by shunting (9) or prosthesis (2) of the renal arteries. In 7 patients percutaneous endovascular angioplasty (PEA) was used. There were neither complications nor lethal outcomes. Arterial pressure normalized in 7 patients and in 9 RVH was better. In the long-term period (at the average within 21 months) 14 patients were followed-up. In 3 patients having perimedial and intimal fibroplasty of RA after PEA restenoses were diagnosed. They had repeated RK. Renal function became better in 1 patient, in 2 patients there was no substantial dynamics. All shunts were passable. The analysis of the data has shown that liquidation of RVH in patients with FMD of RA has inverse dependence on its duration, and decision on the method of RK depends on the type of RA, experience of the specialists, the patients desire included.

[Features of positive personality phenomena in patients with mild cognitive impairment and asthenic syndrome treated with recognan (citicoline)]. / Vliianie primeneniia preparata Rekognan (tsitikolin) na proiavleniia pozitivnykh lichnostnykh kachestv u patsientov s astenicheskim sindromom i legkimi kognitivnymi narusheniiami.

AIM: To assess the formation of positive personality phenomena in patients with mild cognitive impairment and asthenic syndrome during the treatment with recognan (citicoline). MATERIAL AND METHODS: Thirty-eight patients (17 men and 21 women), aged 18 to 45 years (mean age 27.8±12.1 years), with asthenic syndrome with mild cognitive impairment (ICD-10 F06.7) were examined. Patients were divided into two groups: 20 people in the main group and 18 people in the comparison group. The main group received recognan (orally, in solution, 100 mg in 1 ml) for 30 days, the daily dosage of the drug was 0.5 g (5 ml solution). The comparison group did not receive any medications. Adapted methods of positive personality psychology were used: the Fordyce Emotions Questionnaire, the Subjective Happiness Scale (SHS), the Adult Hope Scale (AHS), the Satisfaction with Life Scale (SWLS), M. Atkinson's Scale of Emotional Maturity, the projective technique 'Map of experiences'. The follow-up period was 30 days. All subjects were examined three times (at baseline, 15 and 30 days after treatment). RESULTS AND CONCLUSION: After a month of treatment with recognan, there was an improvement of positive personality traits and a significant decrease in negative experiences, indicating the positive impact of the drug on the formation of positive personality manifestations and compensation for emotional disorders in patients with mild cognitive impairment and asthenic syndrome.

[Percutaneous angioplasty and stenting in treatment of chronic occlusions of renal arteries].

The article presents results of examination and treatment of 14 patients aged from 46 through 79 years with chronic occlusion of one of the renal arteries in each of them and subocclusion of the contralateral renal artery in 13 patients due to atherosclerosis. At first percutaneous intraluminal angioplasty and stenting were performed in 13 out of 14 patients with stenosis of the renal artery. Then at once only percutaneous intraluminal angioplasty of the occlused renal artery was performed in 10 out of 14 patients and within 36 hours on average the implantation of stents in case of patency of the renal artery proper and its branches, according to the findings of control angiography, in these arteries with the optimal result. The percutaneous intraluminal angioplasty and stenting are relatively safe and effective methods of reestablishment of patency of the renal arteries after their chronic occlusion.

[Current treatment options for autonomic, cognitive and emotional disorders in patients with asthenic syndrome treated with recognan (citicoline)]. / Vozmozhnosti lecheniia vegetativnykh, kognitivnykh i émotsional'nykh rasstroistv u patsientov s astenicheskim sindromom pri ispol'zovanii Rekognan (Tsitikolin).

AIM: To study the efficacy of recognan (citicoline) in the treatment of cognitive, emotional, autonomic, asthenic disorders in patients with asthenic syndrome. MATERIAL AND METHODS: Thirty-eight subjects (17 men and 41 women), mean age 27.75±12.05 years, with asthenic syndrome (ICD-10 F48.0 - Neurasthenia) were studied. The sample was randomized into 2 subgroups: the main group (n=20) received oral therapy with recognan (100 mg in 1 ml) for 30 days, while the daily dosage was 500 mg (5 ml). In the control group (n=18), no drug therapy was performed. Patient's state was assessed with a large battery of psychological tests and psychometric scales. The follow-up period was 30 days. All participants were examined three times (initially, in the middle of the study (day 15), in the end of the study (day 30)). RESULTS AND CONCLUSION: Asthenic syndrome was detected in 100% of the patients (total asthenia was noted in 70%, decreased activity in 70%, decreased motivation in 40%, physical asthenia in 45%, mental asthenia in 50%). Recognan (citicoline), used for 2 weeks or one month, has a positive effect on the compensation of autonomic, asthenic cognitive and emotional disorders and increases stress resilience of the patients.

Statistical properties of dynamic speckles formed by a deflecting laser beam.

We analyze statistical properties of dynamic speckles formed when an optically rough surface is illuminated by a fast-deflecting laser beam. The modified space-time correlation function of the light-intensity fluctuations has been introduced to estimate the correlation parameters of a dynamic speckle pattern. Dynamic speckles are considered in their application to range sensing using evaluation of the light-power-modulation frequency of a signal obtained from the integrating photodetector after spatial filtering of the scattered light. Multichannel configuration is suggested to improve the system accuracy. Conditions that should be fulfilled to get uncorrelated responses of photodiode are found. Proper averaging of the multichannel data allows designing a non-interferometric range sensor capable for measuring distance with accuracy of 1 microm during as short time as 1 ms.

[Cognitive and emotional disorders in university students and teachers: the possibility of treatment with recognan (citicoline)]. / Kognitivnye i psikhoémotsional'nye narusheniia u uchashchikhsia i prepodavatelei vuzov: vozmozhnosti korrektsii s pomoshch'iu preparata rekognan (tsitikolin).

AIM: To study the efficacy of recognan (citicoline) in treatment of cognitive, emotional, autonomic, asthenic disorders in university students and teachers. MATERIAL AND METHODS: Fifty-eight subjects (17 men and 41 women), mean age 27.75±12.05 years, including who 42 students and 16 teachers were studied. The sample was randomized into 2 groups (29 each): the main sub-group received oral therapy with recognan for 30 days, while the daily dosage was 500 mg (5 ml). In the control group, no drug therapy was performed. Patient's state was assessed with a large battery of psychological tests and psychometric scales. The follow-up period was 30 days. All participants were examined three times (initially, in the middle of the study (day 15), in the end of the study (day 30)). RESULTS AND CONCLUSION: Asthenic state was detected in 69%, autonomic dysfunction in 82.7%, sleep disorders in 75.8% of subjects. A high level of state anxiety was revealed in 22.2%, trait anxiety in 29.8%, a high level of stress was determined in 6,9%. It is shown that the use of recognan (citicoline) has a positive effect on compensation of autonomic and asthenic disorders, improves cognitive status, corrects psycho-emotional disorders, when used as a short (2 week) and month course.

[The comparative aspects of spatial ecology of lizards exemplified by the toad-headed lizards (Reptilia, Agamidae, Phrynocephalus)].

The possibility of analysis of phylogenetic parameters of the spatial distribution of populations is discussed by an example of the agamid toad-headed lizards (Phrynocephalus). Summarizing both original and published data on the individual home ranges and the relocation of individuals of 30 populations from 12 species showed that differentiation of the type of spatial distribution is weak in toad-headed lizards. This observation confirms the idea that this clade of agamids is phylogenetically young and relatively recently radiated. At the interspecific level, positive correlation between home range size and body size was observed in the studied group. Such spatial parameters, shared by all toad-headed lizards, as relatively large size and weakly structured individual home ranges can be explained by the peculiarities of their reproduction features and their foraging mode. The individual type of space-usage in toad-headed does not fit the traditional scheme dividing all the lizards into the territorial Iguania and the nonterritorial Autarchoglossa.

[Effect of exogenous DNA on the growth of transplantable tumors].

Using transplantable Ehrlich ascites tumor, hepatoma HA-1 and Lewis carcinoma it was shown, that preparations of fragmented genomic DNA can more or less effectively inhibit such tumors as well as the growth of their metastases. Such effects were produced by DNA preparations derived from tissues of mice, both syngeneic or allogeneic to tumor-bearer, as well as from human tissues.