Synthesis, Chemical and Biomedical Aspects of the Use of Sulfated Chitosan.

This work is devoted to the chemical synthesis of sulfated chitosan and its experimental verification in an animal model of early atherosclerosis. The method of chitosan quaternization with sulfate-containing ingredients resulted in a product with a high content of sulfate groups. Implantation of this product into the fascial-muscular sheath of the main limb artery along the leg and thigh in rabbits led to the extraction of cholesterol from the subintimal region. Simplified methods for the chemical synthesis of quaternized sulfated chitosan and the use of these products in a model of experimental atherosclerosis made it possible to perform a comparative morphological analysis of the vascular walls of the experimental and control limbs under conditions of a long-term high-cholesterol diet. The sulfated chitosan samples after implantation were shown to change the morphological pattern of the intimal and middle membranes of the experimental limb artery. The implantation led to the degradation of soft plaques within 30 days after surgical intervention, which significantly increased collateral blood flow. The implantation of sulfated chitosan into the local area of the atherosclerotic lesions in the artery can regulate the cholesterol content in the vascular wall and destroy soft plaques in the subintimal region.

[Variants for conducting situational studies and expertises in forensic medical practice]. / Varianty provedeniya situatsionnykh issledovanii i ekspertiz v sudebno-meditsinskoi praktike.

The aim of the study is to analyze the features of commissioning and conducting the forensic situational studies and expertises and to consider variants for their practical implementation. The importance of a situational analysis when conducting various types of forensic medical expertises is considered. The variants for conducting situational studies and situational expertises in practical work are proposed.

[Forensic, clinical and anatomical aspects of diagnosising diaphragmatic injuries in stab wounds]. / Sudebno-meditsinskie i kliniko-anatomicheskie aspekty diagnostiki koloto-rezanykh povrezhdenii diafragmy.

AIM: To improve the quality of clinical and forensic examination and diagnosis of diaphragmatic injury (DI) from stab wounds by defining the boundaries of the thoraco-abdominal region and the topographic and anatomical characteristics of thoracoabdominal stab wounds. MATERIAL AND METHODS: Based on the results of forensic medical examinations of 81 corpses with stab wounds, the clinical and anatomical characteristics of thoraco-abdominal stab wounds were noted; and in anatomical dissections of 90 cadavers, the boundaries of the intercostal spaces and costophrenic recess were modeled. RESULT: The boundaries of the thoraco-abdominal region, and the location of stab wounds that make the probability of DI particularly high (10-50%) were clarified. A method for diagnosing DI was developed. The method was used to identify DI in 81 out of 411 victims with penetrating stab wounds to the chest and abdomen. The results of the study prove the feasibility of using computed tomography in the examination of living persons with a wound in the thoracoabdominal region. This study can assist in determining the features of a wound channel, in providing a correct assessment of the damage, and in the identification of undiagnosed DI.

[Classification of medical-criminalistic situational examinations]. / Klassifikatsiia mediko-kriminalisticheskikh situatsionnykh ékspertiz.

The aim of this study was to develop a detailed applied classification of medical-criminalistic situational examinations. This paper proposes a classification of importance for practice and research of medical-criminalistic expertise on the reconstruction of events. The basic criteria which determine how the classification of situational examinations is performed are considered.

[The efficiency of the application of the modern computed technologies in the clinical practice and the prospects for the further use of the biomechanical 3D-models in forensic medicine]. / Éffektivnost' ispol'zovaniia sovremennykh komp'iuternykh tekhnologii v klinicheskoi praktike i perspektivy primeneniia biomekhanicheskikh 3D-modelei v sudebnoi meditsine.

To-day, the computer-assisted 3D-technologies for the mathematical simulation of the engineering facilities are extensively used for the purpose of technical calculations in all branches of industry and building. The positive experience gained with the application of the 3D-models finds wide application in the joined investigations on the topical problems of the prosthetic and surgical treatment of bones, teeth, joints, cardiac valves, blood vessels, etc. The objective of the present study was the analysis of the positive experience with the involvement of the specialists in the design and practical application of 3D-models for the solution of problems facing the medical prosthetics and the management of various pathological conditions. Another objective was to discuss the possible prospects for the interdisciplinary collaboration in these fields with a view to improving the quality of expert conclusions in the framework of forensic medical and criminalistics examinations. The data readily available from the official domestic and foreign Internet resources were used for the purpose of the study. The analysis of the published data has demonstrated the obvious advantages of the application of the mathematical 3D-models and the biomechanical studies for the solution of the concrete medico-biological problems. The currently available positive experience gained due to the participation of domestic specialists in biomechanics in the solution of the specific clinical problems gives hope that their collaboration between themselves and with the forensic medical experts will open up the promising prospects for the further investigations of the issues of common interest.

[The commission/comprehensive forensic medical expertise with the participation of the medical professionals]. / Sudebnaia komissionnaia/kompleksnaia ékspertiza s uchastiem vrachei.

The present article deals with the categorization of the forensic medical expertises with respect to their ranking as commission and/or comprehensive ones. The arguments for and against such an approach and interpretation are presented.

[Preimplantation Genetic Diagnosis by Blastocentesis: Problems and Perspectives].

The discovery of cell-free DNA in blastocoele fluid opens new perspectives for the development of preimplantation genetic diagnosis of human chromosomal and genetic diseases. In this review we analyzed the results of the first studies, which made it possible to evaluate the effectiveness of the application of a new source of biological material and showed a high degree of agreement between the results of molecular karyotyping with cell-free DNA and blastocyst cells. The results suggest the possibility of developing a noninvasive method of preimplantation genetic diagnosis, which may open a new round of progress in the field of assisted reproductive technologies and the genetics of early stages of human ontogenesis.

MOLECULAR KARYOTYPING BY USING CELL-FREE DNA FROM HUMAN BLASTOCOELE FLUID, EMBRYOBLAST AND TROPHOBLAST CELLS.

In the present study, we have carried out a comparative analysis of molecular karyotypes of cell free DNA from blastocoele fluid, trophectoderm and inner cell mass of human blastocysts at the preimplantation stages of development, using the comparative genomic hybridization. Different types of chromosomal abnormalities distribution between trophectoderm, inner cell mass and blastocoele fluid were identified. It was first shown that the source of cell free DNA in the blastocoele fluid may arise from the cells of inner cell mass in addition to the trophectoderm cells, which are traditionally used in preimplantation genetic diagnosis of chromosomal aberrations. These results support the promising of blastocentesis as a new technology of preimplantation genetic diagnosis, which provides the possibility of integral analysis of chromosome abnormalities in different cells of the blastocyst.

[Sperm DNA fragmentation: association with semen parameters in young men].

AIM: Abnormal sperm DNA integrity is an important risk factor for male infertility. The aim of this work was to examine sperm DNA fragmentation in a cohort of young male volunteers (n=111; age 21.0+/-0.2 years) from the general population and establish the association between the level of sperm DNA fragmentation and sperm functional parameters. MATERIALS AND METHODS: Sperm DNA fragmentation index (DFI) was determined by SCSA (sperm chromatin structure assay) using flow cytometry. Standard semen parameters (concentration, motility, and morphology) were evaluated according to the WHO guidelines (2010). RESULTS: and conclusions. In the study cohort, 79.0%, 12.4% and 8.6% of men had normal (DFI<15%), borderline (15 less or equal DFI<27%) and high (DFI more or equal 27%) levels of fragmentation, respectively. Men with impaired spermatogenesis had greater IDF values (14.53+/-1.43%) than men with normal semen parameters (8.88+/-0.77%, p<0.05). There was a statistically significant negative correlation between IFD and ejaculate concentration (r=-0.21, p<0.05), fractions of mobile (r=-0.41, p<0.05) and morphologically normal sperm (r=- 0.34, p<0.05). Testing sperm DNA fragmentation using SCSA technique can be employed in epidemiological studies of male fertility.

[THE OPTIMAL CONDITIONS OF STORAGE OF SPERMATOZOA FOR ANALYSIS OF DNA FRAGMENTATION].

The analysis of fragmentation of DNA of spermatozoons using technique of flow cytometry to evaluate male fertility more and more often begins to be applied in clinical diagnostic. However, development of optimal protocol of storage and preparation of spermatozoons for analysis still is at the stage of experimental elaboration. The studv was carried out to analyse effect of different conditions of preparation of ejaculate for adequate evaluation of index of fragmentation of DNA of spermatozoons using sperm chromatin structure assay technique. The sampling consisted of 20 patients of the Krasnoyarsk center of reproductive medicine. The sperm chromatin structure assay technique was applied to evaluate index of fragmentation of DNA of spermatozoons in fresh native ejaculate and after storage of spermatozoons under different temperature (37, 25 and 4 degrees C) and duration (1-2 and 1-3 days) and conditions of storage (-20 or -70 degrees C) of frozen spermatozoons (as native ejaculate or in TNE-buffer). It is demonstrated that index of fragmentation of DNA of spermatozoons has no significant alterations in ejaculate stored under 4 degrees C during 48 hours. In case of storage of ejaculate under 25 or 37 degrees C index of fragmentation of DNA of spermatozoons significantly increases already after first day of storage. The incubation of ejaculate under 37 degrees C results in increasing of index of fragmentation of DNA of spermatozoons already after first hour. The individual differences are established related to degree of increasing of index of fragmentation of DNA of spermatozoons because of impact of studied temperatures of ejaculate incubation. The storage of spermatozoons under temperature of - 20 and -70 degrees C in native ejaculate or in TNE-buffer has no effect of index of fragmentation of DNA of spermatozoons with measurement during 1-2 hours. Therefore, storage and transportation of native ejaculate under 4 degrees C during 1-2 days or in frozen condition under temperature of -20 degrees C or -70 degrees C can be recommended for adequate evaluation of level of fragmentation of DNA of spermatozoons.

[Molecular Karyotyping of Cell-Free DNA from Blastocoele Fluid as a Basis for Noninvasive Preimplantation Genetic Screening of Aneuploidy].

The discovery of DNA fragments in the blastocoele fluid is promising for the development of new noninvasive methods for the preimplantation genetic diagnosis of chromosomal diseases. However, to date there are no data confirming the concordance between the molecular karyotype of cell-free DNA from blastocoele fluid and the blastocyst cells per se. This paper reports on this concordance according to the results of molecular-cytogenetic analysis of the chromosomal set with the use of comparative genomic hybridization.

[The determination of the ballistics of a hunting rifle loaded with a Poleva-3 bullet].

Various approaches are considered to determine the shooting range of a hunting rifle loaded with a Poleva-3 bullet from the specific features of gunshot injuries inflicted by container-type Poleba-3 bullets for hunting rifles.

[Level of DNA fragmentation in human sperm cells in varicocele and prostatitis].

Varicocele and prostatitis are the most common andrological diseases, which may be accompanied by a decrease in the production of sperm cells, the deterioration of their quality and increased risk of infertility. This work was aimed to the evaluation of sperm DNA fragmentation index (DFI) and main indices of sperm fertility (concentration, motility and morphology), and the relationship between these parameters in the men of active reproductive age suffering from prostatitis or varicocele. Assessment of sperm DNA fragmentation was performed by SCSA (sperm chromatin structure assay) using flow cytometry; sperm parameters were evaluated according to WHO recommendations. It was shown that men with prostatitis (n = 9) and varicocele (n = 22) had significantly higher DFI compared with men in the control group (n = 22). Negative influence of these diseases on the concentration and the percentage of motile sperm cells in the ejaculate was revealed. These data suggest that the deterioration in the quality of semen in varicocele and prostatitis may be caused not only by pathospermia, but also, at least partially, by violation of the integrity of the sperm DNA. Evaluation of sperm DNA fragmentation can be recommended for use in laboratory diagnostics for prediction of fertility in infertile men.

[A method for the determination of spatial orientation of wound canals caused by gunshot bullets to be applied in corpse examination].

A new approach to the determination of spatial orientation of wound canals caused by gunshot bullets is proposed and substantiated; its application for the purpose of forensic medical expertise is described. The direction of penetrating wound canals in three dimensions was determined by measuring the distance of the input and output wound holes from the level of the soles, anterior and posterior midlines, and the posterior body surface; the results of the measurements were used to construct triangles. The parameters thus obtained allowed the spatial orientation of the wound canals to be determined in three dimensions for the subsequent judgment about the relative direction of the gun barrel with respect to the human body at the time of the shot and the objective expert conclusion concerning the circumstances of firearm-related injuries.

[Allocation of precursor germinal cells from the tissue of the human ovaries].

The article present the results of a study on the allocation of precursor germinal cells from tissue of human ovaries obtained during various gynecological operations. Two series of cell culture were isolated from tissue of tunica albuginea. The cell lines were identified and characterized by PCR012-method, immunocytochemical staining and histological examination. The study provided data reliably confirming the presence of precursor cells in the ovarian tissue of women in their reproductive years.

Mouse hepatocytes retain the expression of the main differentiation markers during culturing on collagen-chitosan matrices.

Mouse hepatocytes cultured on artificial 3D collagen-chitosan biopolymer matrices retained the expression of hepatocyte markers for 14 days.

Cytotoxic activity of endometrial immunocompetent cells determines the outcome of embryo implantation.

The data on the formation of blastocysts in culture during therapeutic cycles of extracorporeal fertilization and on cytotoxic index of the endometrium in women with different clinical pathogenetic variants of sterility are presented. Analysis of correlations of these parameters with the incidence of effective implantation and pregnancy revealed a strict correlation between cytotoxic activity of endometrial immunocompetent cells and human embryo implantation.

[The forensic medical assessment of the results of a study of laryngeal injuries in blunt trauma to the neck]. / Sudebno-meditsinskaia otsenka rezul'tatov issledovaniia povrezhdenii gortani pri tupoi travme shei.

Presents the methodological principles and succession of examination of the basic formations of the larynx in cases with blunt injuries of the neck. Describes variants of anatomic structure of the sublingual bone and laryngeal cartilages influencing the morphology of injuries thereof. Offers differential diagnostic criteria of various mechanisms of fractures of the basic formations and recommendations on medical criminological assessment of laryngeal injuries.