[Non-transecting anastomotic urethroplasty for strictures of the bulbar urethra. Systematic review].

The results of using non-transecting anastomotic urethroplasty in men with bulbous urethral strictures are presented in the review. A total of 25 original publications were found, including 20 foreign and 5 Russian articles. The studies included from 1 to 358 patients who underwent anastomotic urethroplasty without transection of the corpus spongiosum (average number of patients in a study was 54). Etiological factors were indicated in 17 articles. Most studies (10 out of 17) indicated idiopathic etiology as the predominant one. There was no correlation between the results of the procedure and the etiology of urethral stricture. The mean length of urethral stricture in the vast majority of studies was less than 2 cm, and only in a few studies it was larger, with a maximum mean value of 3.9 cm. Postoperative complication rates were reported in 20 studies and ranged from 0% to 23.9% within one study (median 8.4%). In general, mild complications occurred, corresponding to category I-II according to the Clavien-Dindo classification. The incidence of erectile dysfunction was evaluated in 18 studies and ranged from 0% to 23% (average value of 6.5%). The success of non-transecting anastomotic urethroplasty averaged 94.7% (82-100%) with a median postoperative follow-up of 24.5 months (3-150 months). In 9 out of 25 studies, an additional comparison with transecting technique was done. In 6 studies, the superiority of the non-transecting technique in terms of treatment success and preservation of sexual function was found. The obtained results showed the high efficiency and safety of non-transecting anastomotic urethroplasty in case of short strictures of the bulbous urethra.

[Giant prostate stones: case report].

Small prostate stones are often found incidentally during clinical and radiological examination. Large stones, however, also may develop, replacing completely the prostate tissue and causing various symptoms. Such a large stones are commonly formed due to chronic urine reflux. There are 20 publications in the literature devoted to the patients with giant prostate stones. Open as well as endoscopic procedures can be performed. In our clinical case both approaches were done simultaneously. Such tactic was chosen in order to carry out a single-stage intervention, immediately solving two problems, i.e., urethral stricture and a giant prostate stone.

[Prospects of Genetically Encoded Flim Indicators for the Quantitative Assessment of Intracellular Parameters].

A significant number of genetically encoded indicators based on fluorescent proteins that allow detecting changes in various parameters: membrane potential shift, pH, concentrations of hydrogen peroxide, lactate, pyruvate, NAD+/NADH, ATP, calcium cations, etc. have been created. Some of them (for example, indicators of calcium cations and hydrogen peroxide) are successfully used by numerous groups of researchers in experiments in vivo. The fundamental limitation of most fluorescent indicators is their ability to only qualitatively assess the change in the parameter of interest in the cell. The nature of this limitation is that it is difficult to compare the absolute value of the measured parameter and the detected fluorescent signal for various reasons. Quantitative measurements impose an additional requirement on the indicator signal: it should not depend on the technical features of the experiment (such as the level of protein expression, the type and power of the excitation source, a set of fluorescent filters, etc.). The fulfillment of this requirement may be associated with the detection of the indicator signal in the time domain, where the main characteristic of the signal is the fluorescence lifetime. The fluorescence lifetime is an absolute value, insensitive to changes in the intensity of the excitation source, detection settings, losses in the optical path or sample, changes in the concentration of the fluorophore in the sample, photobleaching. Thus, when calibrating an indicator in the time domain, a specific value of the measured parameter is associated with a specific value of the fluorescence lifetime. The time-resolved imaging mode imposes its own limitations on the properties of the indicator's fluorescent core. Thus, the lifetime of its fluorescence should change in proportion to the value of the measured parameter, which is not characteristic of any fluorophore. In this work, we make a brief review of the principles of detection of the fluorescent signal of genetically encoded indicators, focusing on current approaches related to the analysis of time-resolved data.

Monitoring of the heavy-ion beam distribution using poly- and monochromatic x-ray fluorescence imaging.

In this work, the first proof of the principal of an in situ diagnostics of the heavy-ion beam intensity distribution in irradiation of solid targets is proposed. In this scheme, x-ray fluorescence that occurs in the interaction of heavy-ions with target atoms is used for imaging purposes. The x-ray conversion to optical radiation and a transport-system was developed, and its first test was performed in experiments at the Universal Linear Accelerator in Darmstadt, Germany. The Au-beam intensity distribution on thin foils and Cu-mesh targets was imaged using multiple x-ray pinholes (polychromatic imaging) and 2D monochromatic imaging of Cu Kα radiation by using a toroidally bent silicon crystal. The presented results are of importance for application in experiments on the investigation of the equation of states of high energy density matter using high intensity GeV/u heavy-ion beams of ≥1010 particles/100 ns.

[Thioredoxin in fibroblasts of human dermis in the process of aging.]

The aim of this work was to examine the content of thioredoxin in fibroblasts of human dermis from the development until deep aging (from 20 weeks of pregnancy until 85 years old), and defining of a role of thioredoxin in age-dependent changes in the number of fibroblasts in the dermis. Thioredoxin, proliferating cells nuclear antigen (PCNA), marker of fibroblasts vimentin were detected with indirect immunohistochemical technique. Results showed that portion of fibroblasts with positive staining for thioredoxin in the dermis is increased from 20 weeks of pregnancy until 85 years old. Most expressed age related increase in portion of thioredoxin positive dermal fibroblasts (more than 9 times) is present form birth until 20 years as compared to antenatal period. General number and percent of PCNA positive fibroblasts in dermis are decreased with age with more expressed changes until 40 years old. Correlation analysis showed that age dependent decrease in the number of fibroblasts and their proliferative activity is significantly associated with increase in thioredoxin positive fibroblasts in dermis. Results allow to suggest that thioredoxin play a role in age dependent decrease in the number of fibroblasts and their proliferation in human dermis.

[Perineal urethrostomy: results, clinical, statistical and terminological aspects].

The analysis of the results of perineal urethrostomy for urethral stricture in men for the period from 2000 to 2020 is presented in the article. We identified 29 original articles devoted to this technique, including 4 Russian and 25 foreign publications. In two studies, there were more than 100 patients, four studies included from 51 to 77 men, and in remaining studies less than 50 patients were analyzed. Complete data on all pre- and postoperative aspects of perineal urethrostomy were provided only in a few studies. In the overwhelm of cases, perineal urethrostomy was performed after one or several unsuccessful reconstructive procedures and only in few patients it was done prior to various types of urethroplasty. It was not possible to establish any association between etiological factors and localization of urethral stricture among these patients. The incidence of pan-urethral stricture ranged from 28.6% to 100%. Only in one study the average stricture length was 2 cm, while in all other studies it was 5 cm or more. Postoperative follow-up was on average 14 to 62 months. Treatment success with perineal urethrostomy ranged from 0% to 100%, but in most studies it was 75% or more. The most common perineal urethrostomy technique used included Blandy technique, and Johanson and 7-flap techniques.

Orientation Distribution of Molecules in a Smectic Liquid Crystal with a Distorted Director Geometry.

Quantitative determination of the molecular orientation distribution function in samples of liquid crystals with a complex director geometry was performed using the numerical simulation of electron paramagnetic resonance (EPR) spectra of the spin probes in a liquid-crystalline medium. To achieve the quantitative agreement of experimental and simulated EPR spectra, the hierarchy of the orientation order was explicitly taken into account, namely, the local ordering of liquid crystal molecules by the mean-field potential of surrounding molecules, and the partial disordering of local directors within the sample. The samples under study are planar and twist LC cells with liquid crystal 8CB cooled from the nematic into the smectic A phase in the magnetic field. The presence of the magnetic field perpendicular to the cell director leads to distortion of the orientation of the liquid crystal in the cell. The spin probe technique was successfully employed for the reliable measurement of orientation distribution functions of the low nonorthorhombic symmetry. Orientation order parameters up to 12th rank were measured, including nonaxial and nonorthorhombic order parameters. It is shown that the presence of several contradicting aligning forces leads to the tilt of the preferential director toward the direction, which is a compromise between the orienting forces.

Molecular Tools for Targeted Control of Nerve Cell Electrical Activity. Part I.

In modern life sciences, the issue of a specific, exogenously directed manipulation of a cell's biochemistry is a highly topical one. In the case of electrically excitable cells, the aim of the manipulation is to control the cells' electrical activity, with the result being either excitation with subsequent generation of an action potential or inhibition and suppression of the excitatory currents. The techniques of electrical activity stimulation are of particular significance in tackling the most challenging basic problem: figuring out how the nervous system of higher multicellular organisms functions. At this juncture, when neuroscience is gradually abandoning the reductionist approach in favor of the direct investigation of complex neuronal systems, minimally invasive methods for brain tissue stimulation are becoming the basic element in the toolbox of those involved in the field. In this review, we describe three approaches that are based on the delivery of exogenous, genetically encoded molecules sensitive to external stimuli into the nervous tissue. These approaches include optogenetics (Part I) as well as chemogenetics and thermogenetics (Part II), which are significantly different not only in the nature of the stimuli and structure of the appropriate effector proteins, but also in the details of experimental applications. The latter circumstance is an indication that these are rather complementary than competing techniques.

Molecular Tools for Targeted Control of Nerve Cell Electrical Activity. Part II.

In modern life sciences, the issue of a specific, exogenously directed manipulation of a cell's biochemistry is a highly topical one. In the case of electrically excitable cells, the aim of the manipulation is to control the cells' electrical activity, with the result being either excitation with subsequent generation of an action potential or inhibition and suppression of the excitatory currents. The techniques of electrical activity stimulation are of particular significance in tackling the most challenging basic problem: figuring out how the nervous system of higher multicellular organisms functions. At this juncture, when neuroscience is gradually abandoning the reductionist approach in favor of the direct investigation of complex neuronal systems, minimally invasive methods for brain tissue stimulation are becoming the basic element in the toolbox of those involved in the field. In this review, we describe three approaches that are based on the delivery of exogenous, genetically encoded molecules sensitive to external stimuli into the nervous tissue. These approaches include optogenetics (overviewed in Part I), as well as chemogenetics and thermogenetics (described here, in Part II), which is significantly different not only in the nature of the stimuli and structure of the appropriate effector proteins, but also in the details of experimental applications. The latter circumstance is an indication that these are rather complementary than competing techniques.

Rotational Dynamics of Nitroxide Biradical in Room-Temperature Ionic Liquids Measured by Quantitative Simulation of EPR Spectra.

Temperature dependences of electron paramagnetic resonance (EPR) spectra of an imidazoline nitroxide biradical spin probe in a series of room-temperature ionic liquids in the temperature range 124-390 K have been quantitatively simulated. The unusual asymmetric EPR spectrum shape previously observed in these systems [Kokorin et al., Appl. Magn. Res. 48 (2016) 287] is shown to originate from anisotropic rotational diffusion of the probe molecule. All experimental spectra were quantitatively reproduced in simulation using a unified set of geometrical and magnetic parameters of the spin probe, which were found to be fully consistent with the biradical geometry obtained from density functional theory calculations. Temperature dependences of rotation diffusion coefficient of the probe characterize the molecular mobility of the ionic liquid, whereas the temperature dependences of the spin-exchange integral J and of the isotropic hyperfine interaction constant, aN, are shown to reflect the intramolecular conformation motions of the biradical probe.

Molecular Principles of Insect Chemoreception.

Chemoreception, an ability to perceive specific chemical stimuli, is one of the most evolutionarily ancient forms of interaction between living organisms and their environment. Chemoreception systems are found in organisms belonging to all biological kingdoms. In higher multicellular animals, chemoreception (along with photo- and mechanoreception) underlies the functioning of five traditional senses. Insects have developed a peculiar and one of the most sophisticated chemoreception systems, which exploits at least three receptor superfamilies providing perception of smell and taste, as well as chemical communication in these animals. The enormous diversity of physiologically relevant compounds in the environment has given rise to a wide-ranging repertoire of chemoreceptors of various specificities. Thus, in insects, they are represented by several structurally and functionally distinct protein classes and are encoded by hundreds of genes. In the current review, we briefly characterize the insect chemoreception system by describing the main groups of receptors that constitute it and putting emphasis on the peculiar architecture and mechanisms of functioning possessed by these molecules.

Apoptotic Bodies of Cardiomyocytes and Fibroblasts - Regulators of Directed Differentiation of Heart Stem Cells.

We studied the effects of apoptotic bodies of cardiomyocytes (ApBc) and fibroblasts (ApBf) on myocardial regeneration and contractility in rats and the dynamics of RNA concentrations in cardiomyocytes and fibroblasts at different stages of apoptosis. ApBc increase the contractility of rat myocardium, while ApBf reduce it. ApBc stimulate the development of clones of cardiomyocyte precursors in the myocardium, while ApBf stimulate the formation of endothelial precursor clones. In doxorubicin cardiomyopathy, ApBc, similar to the reference drug (ACE inhibitor) improve animal survival, while ApBf produce no such effect. RNA concentrations in cardiomyocytes and fibroblasts before apoptosis and at the beginning of cell death significantly differed, while in apoptotic bodies of these cells, it was practically the same. It has been hypothesized that RNA complex present in ApBc and ApBf represents an "epigenetic code" of directed differentiation of cardiac stem cells.

[Oncological safety of nerve-sparing radical prostatectomy: evaluation of histopathological outcomes and recurrence-free survival].

OBJECTIVE: To evaluate histopathological outcomes and biochemical recurrence (BCR) free survival in patients after nerve-sparing radical prostatectomy (nsRP). MATERIALS AND METHODS: Prospective study group comprised 313 patients who underwent uni- or bilateral nsRP from 2014 to 2018; control group included 592 patients with clinically localized prostate cancer who underwent non-nsRP from 2014 to 2018. Mann-Whitney U-test was performed to assess continuous variables; chi-squared test was used for comparative analysis of categorical data. BCR free survival was evaluated with Kaplan-Meier method, log-rank test was used to compare survival outcomes. RESULTS: Adverse histopathological findings were lower in the study group: extracapsular extension was found in 9,4% and 18,75% (p<0,001), grade group upgrade in 23% and 29,3% (p=0,04), positive surgical margins (PSM) in 15% and 22,1% (p=0,01). Subanalysis according to cancer risk groups showed lower PSM rates in high-risk patients (15,6% and 30,3%, p=0,017) and tendency for higher PSM rates in low-risk patients in the study group with no significant difference (12,6% and 7%, p=0,16). BCR free survival after 12 months was 100% and 88,2%, after 20 months - 92,3% and 86,4%, p=0,04. PSM ratesin the study group were notsignificantly different depending on the type of surgery: 13,9% in robotic-assisted approach and 15,4% in retropubic approach, p=0,75. CONCLUSIONS: Relatively favorable histopathological outcomes and BCR free survival can be achieved after nsRP. However, obtained results could not be considered optimal and clearly indicate the need for further improvement of preoperative planning and intraoperative quality control of surgical treatment.

[Variability of Fragments of Nuclear Brca1 Gene, Exon 11, and Mitochondrial Cox1 Gene in House Mice Mus musculus].

To clarify genetic differences between subspecies of the house mouse Mus musculus, their distribution, and hybridization, we first conducted a comparative analysis of variability of nucleotide sequences of fragments of the nuclear gene Brca1, exon 11 (2331 bp), and mitochondrial gene Cox1 (1260 bp) in 40 house mice from West and East Europe, Transcaucasia, Siberia, and Central and South Asia. Brca1 genotypes were divided into five main groups, which differed in a number of fixed substitutions. Genotypes of each group are characteristic for the certain geographical region and the following subspecies: M. m. musculus, M. m. domesticus, M. m. castaneus, and M. m. wagneri together with M. m. gansuensis; a fifth group corresponds to an unidentified subspecies or a distinct genetic form of M. musculus from India (Sikkim State). Besides the homozygous specimens, we revealed mice, which were heterozygous for all diagnostic loci simultaneously; these specimens were determined as hybrid. Hybrid mice were mainly found in the zones of contact of subspecies, but in some cases, quite far from one of the parent subspecies (possibly, due to transportation). In two hybrid mice (from Bakhtiari Province of Iran and Transbaikalia of Russia), unique Brca1 haplotypes were detected. It cannot be ruled out that, at least partly, they may be characteristic of the M. m. bactrianus and M. m. gansuensis subspecies, respectively. Thus, the results of the study showed that the nuclear Brca1 gene is a promising molecular genetic marker for the analysis of variability, differentiation, and hybridization of house mice as well for subspecific identification of M. musculus specimens. Despite more rapid evolution of the Cox1 gene, it is not well suited for discrimination of M. m. musculus, M. m. wagneri, M. m. gansuensis specimens and Transcaucasian representatives of M. m. domesticus due to introgression and long-term maintenance of foreign mitochondrial DNA in populations. However, Cox1 gene analysis (along with the diagnostics of animals by nuclear DNA) may be useful for estimation of population differences in M. m. castaneus and M. m. domesticus subspecies.

[Iron deficiency anemia in elderly and senile patients.]

Anemic syndrome is common in 1/3 of the population, including iron deficiency anemia - in 1,5 billion people. Geriatric patients are one of the main risk group for anemia. Iron deficiency and iron deficiency anemia lead to a decrease in quality of life, an increase in morbidity and mortality, what requires timely diagnosis and treatment. The diagnostic algorithm includes the analysis of iron metabolism, inflammation markers and instrumental tests to verify the cause of anemia. Modern oral and parenteral iron preparations are used for treatment under control of blood indexes and iron metabolism parameters.

Allosteric regulation of the ribosomal A site revealed by molecular dynamics simulations.

The A site of the ribosome, which determines binding and orientation of a new amino acid residue for the peptidyl transferase reaction, was found to occupy two different conformational states upon the molecular dynamics (MD) simulations study of the 70S E. coli ribosome. One of the states, defined as "inactive", appeared in trajectories with E-tRNA, mutations A2531U and UU2492-3C, which are known to decrease the A site affinity to the tRNA. This conformational transition was found to be allosterically connected with conformational alterations in different sites of the macromolecular complex, including the E site of the ribosome and intersubunit bridge B7a located near the E site. The MD simulations of the ribosomes with the A2531U and UU2492-3C mutations known to decrease the A-tRNA retention in the ribosome, demonstrated partial switching of the 16S and 23S rRNA conformation towards its characteristic one in the P/P, E/E state.

[Non-transecting anastomotic urethroplasty].

The review analyzes the results of using non-transecting anastomotic urethroplasty in men with urethral strictures. Identified 14 original studies using this technique: 13 foreign and 1 Russian. In total, this technique was applied in 704 patients. This technique was used in 85% of cases with bulbar urethral stricture, in 15% - with posterior stricture.The average length of the urethral stricture in all studies was less than 2 cm, except for two works, where the average stricture length was 2.3 cm and 3.9 cm, respectively. This technique is equally successfully applied in all etiological variants of urethral stricture. The incidence of postoperative complications in all studies using non-transecting anastomotic urethroplastyaveraged 13.7%.Postoperative complications were mostly mild and corresponded to G1 according to Clavien-Dindo classification.According to 10 out of 14 studies that evaluated the effect of surgical treatment on the occurrence of erectile dysfunction (ED), the incidence of ED de novo was on average 13.4%. The success of treatment with the use of non-transecting anastomotic urethroplastyaveraged 95% (82-100%) with a median postoperative follow-up of 27 months (6-64 months). In 4 out of 14 studies, an additional comparison was transecting versus non-transecting techniques. Non-transecting technique was not inferior to transecting technique by any criterion, but, on the contrary, exceeded it in a number of indicators, including the dynamics of sexual function after surgery.On the basis of available studies on the use of non-transecting anastomotic urethroplasty, this technique should be recognized as a highly efficient and reasonably safe method for treating short urethral strictures and recommended for widespread clinical use.

Sensors for Proteolytic Activity Visualization and Their Application in Animal Models of Human Diseases.

Various sensors designed for optical and photo(opto)acoustic imaging in living systems are becoming essential components of basic and applied biomedical research. Some of them including those developed for determining enzyme activity in vivo are becoming commercially available. These sensors can be used for various fluorescent signal detection methods: from whole body tomography to endoscopy with miniature cameras. Sensor molecules including enzyme-cleavable macromolecules carrying multiple quenched near-infrared fluorophores are able to deliver their payload in vivo and have long circulation time in bloodstream enabling detection of enzyme activity for extended periods of time at low doses of these sensors. In the future, more effective "activated" probes are expected to become available with optimized sensitivity to enzymatic activity, spectral characteristics suitable for intraoperative imaging of surgical field, biocompatibility and lack of immunogenicity and toxicity. New in vivo optical imaging methods such as the fluorescence lifetime and photo(opto)acoustic imaging will contribute to early diagnosis of human diseases. The use of sensors for in vivo optical imaging will include more extensive preclinical applications of experimental therapies. At the same time, the ongoing development and improvement of optical signal detectors as well as the availability of biologically inert and highly specific fluorescent probes will further contribute to the introduction of fluorescence imaging into the clinic.

Spin Probe Determination of Molecular Orientation Distribution and Rotational Mobility in Liquid Crystals: Model-Free Approach.

A model-free approach for simulation of EPR spectra of nitroxide spin probes in liquid-crystalline materials was suggested and used to obtain parameters of molecular orientation and rotational mobility. The developed method is based on experimental recording and numerical simulation of the angular dependence of EPR spectra, which is shown to be much more informative in comparison with a single EPR spectrum. Quantitative spectral simulations considering both local orientational ordering and distribution of local directors in the sample were used for discrimination of models of rotational mobility and orientational alignment. The method was applied for detailed quantitative characterization of axial, orthorhombic, and low-symmetry non-orthorhombic molecular orientation distributions. It is shown that the ordinarily used model of rotational diffusion in a mean-field potential is suitable for the description of molecular mobility and orientational ordering only for relatively low sample temperatures and low-mobility probe molecules with large sizes. In cases of high molecular mobility, the more realistic jump mechanism of molecular moves can be approximately described as quasi-librations. For ordered liquid crystals it was found that mostly the order parameters up to rank 12-14 are essential and easily determined. When well-aligned materials are described, the order parameters up to 18th rank or even higher become meaningful. Both molecular and sample biaxiality is analyzed and quantitatively characterized. The local molecular ordering and sample orientational alignment are quantitatively characterized separately.

[Characteristics of the Renin-Angiotensin-Aldosterone System in Carriers of Polymorphic Variants of the VDR Gene With Arterial Hypertension and Obesity].

AIM: to study the renin-angiotensin-aldosterone-system (RAAS) in patients with polymorphic variants of the vitamin D receptor gene (VDR), arterial hypertension (AH) and obesity. MATERIALS AND METHODS: We included in this study 98 patients with stage II-III AH and obesity (33 men, 65 women, mean age 61.1±9.9 years and BMI 43.3±5.0 kg/m2). The VDR gene study included genotypes TaqI, BsmI, FokI. After identification of TaqI, BsmI, FokI VDR gene sites we carried out measurements of levels of RAAS markers. RESULTS: We revealed an associative relationship between the presence of allele G of the BsmI site, C/C homozygosity of FokI site of VDR gene and presence of AH in patients with obesity. An increase of renin level was noted only in the homozygous T/T genotype of FokI, no differences in the concentration of aldosterone and angiotensin were detected between any of the studied genotypes. CONCLUSION: Polymorphic alleles and genotypes of the VDR gene can be used as predictors of AH development in obese patients with subsequent characterization of the state of the RAAS system.