As. Varukha K.,   MD, Prof. Babkina O.

              National Medical University O.O. Bogomolets  Kyiv, Ukraine

 

 THE QUESTION OF DETERMINING THE ORIGIN OF MENSTRUAL BLOOD PROSTAGLANDIN F2A THE CONTENTS

 

Introduction: In recent years, actively studied microparticles, isolated tissue cells of the body, spots like the blood that remains as overlays on the body, clothes in the genital area affected with sexual crimes. Sometimes on physical evidence can detect large-size pieces of fabric, often out microscopic particles and even isolated single cells. Along with determining the presence of blood in the wake of evidence, determining its species, group and gender, regional origin of blood, and quite often the question arises whether the menstrual blood. To date, the contemporary forensic practice is not developed enough dostavirnyh methods of determining the origin of menstrual blood stains, and therefore the question that often is of fundamental importance for the investigation usually remains unanswered. Therefore, the definition of the regional origin of the individual components of mixed stains (blood) is an important and at the same time a challenge. To address issues of regional origin components mixed stains (blood) proposed to define different criteria: the presence of vaginal epithelial cells, columnar epithelium of the uterus, as well as a large number of bacteria - cocci, bacilli; the presence of particular substances - menotoksina; fibrinolytic activity, especially the macro - and trace element composition, etc. [1-6]. For example, for a comparative study were subjected to blood stains at nemenstrualniy bleeding from the genital tract of women and bleeding from other sources [5,6]. However, research presented by different authors do not go beyond scientific inquiry, and therefore modern forensic practice is not any reliable method of determining menstrual blood origin.

Objective: The aim of the study was to develop criteria for the menstrual blood, according to the Prostoglandin origin, namely prostaglandin F2alfa.

Material and Methods: The object of the study was forced on gauze menstrual blood taken from 20 women aged 18 to 40 years with normal menstrual cycle, which is the time the study was stored in dry form under normal conditions. In order to identify the origin of menstrual blood was applied our method preparative separation and systematic course of analysis Prostoglandin obtained by biosynthesis proposed RV Bobylev et al [4]. Research object filled with alcohol 960 C in an amount of 25 ml and extracted for 24 hours. In a test tube with 10 ml of extract fill up the buffer of pH 7,0 and intensively shaken. Alcohol evaporated in a vacuum at 400 C, then acidified 3% Murashova acid to pH 4,3. Three times extracted with chloroform 3 ml, intensely shaken and centrifuged. The combined chloroform extracts were evaporated in a vacuum at 400 C. The balance contributed 0.5 ml of ethanol and stirred. Qualitative analysis sulfuric fraction of prostaglandins performed by chromatography on plates Silufol RUV 254 in system chloroform - methanol - acetic acid - water in the ratio 90: 8: 1: 0.8. Upon reaching the front end of the plate, it was removed from the chamber and dried in air after placed in Filling amount of iodine vapor spots for display of prostaglandins. Control standard solution was F2alfa prostaglandin (Enzaprost F).

Relationship with academic programs, plans, themes.

Article is part of a comprehensive research on the topic: "Forensic installation menstrual blood origin", which is performed at the Department of Forensic Medicine of the National Medical University. OO Bogomolets and has state registration number 0114U007149.

Results: In the course of our research when deciding on the definition of origin menstrual blood are important research Khimich regulation of the menstrual cycle hormonal and menstrual blood composition origin. It is well known that the intensity of menstruation determined not only by the structure of the endometrium until its rejection and platelet aggregation, but also the contractile activity of the myometrium and arterioles. These processes are closely related with the degree of synthesis and degradation of prostaglandins. Prostaglandins represent a special class of biologically active compounds (unsaturated hydroxylated fatty acids), which are found in almost all body tissues. Prostaglandins are synthesized inside the cell and released the same cells to which they apply. Because cell called prostaglandins hormones. The formation of prostaglandins in the endometrium is controlled by estrogen and progesterone, which provides braking action. In connection with the fact that the mechanism of inhibitory effect of progesterone on prostaglandin synthesis is not fully understood, we believe that an important role in this process is the inhibition of phospholipase A2 and phospholipase C, which provide release from phospholipids arachidonic acid, the availability of which is considered to be a limiting factor in biosynthesis of prostaglandins. It is known that under the influence of progesterone decreases prostaglandin synthetase activity and increased activity of 15-hydroxy-prostaglandin dehydrogenase, which converts prostaglandin into inactive metabolites. Estradiol, on the contrary, stimulates the secretion of prostaglandin E2 F2alfa and prostaglandin in the endometrium. One of the main mechanisms for increasing the formation of prostaglandins is influenced by estrogen stimulation of phospholipase A2 activity which significantly increased in the endometrium during the proliferative phase of the cycle in parallel with the increase in secretion of estradiol. In addition, in vitro studies showed that estradiol enhances prostaglandin synthetase activity in human endometrium. The combined action of estradiol and progesterone during the menstrual cycle causes the complex picture of prostaglandin synthesis depends on changing concentrations of both hormones and their relationship. Revealed that estradiol stimulates the secretion of prostaglandin E2 F2alfa and prostaglandin in the endometrium, elongated in the proliferative phase of the cycle. By itself, progesterone inhibits prostaglandin synthesis, but increases the sensitivity of the endometrium to the action of estradiol. Falling levels of progesterone in lyuteolyzi probably reduces its inhibitory effect on phospholipase activity and prostaglandin - synthetase and leads to stimulation of prostaglandin secretion before menstruation In the course of our research we took as a basis the availability in all the sites of prostaglandin F2alfa as an indicator of the presence of menstrual blood. As a result of our studies five objects prostaglandin F2alfa standard solution (Enzaprost F) in the chromatogram spots were detected absorption purple with five factions height 40 mm, 43mm, 48mm, 51mm and 53 mm. The length of the Front (start lifting up the chromatographic eluent layer by capillary forces) was 100 mm. Upon reaching the front line height sufficient for analysis, we investigated plate removed and dried. Distance, which is the front line, counted from the starting line, which is sufficient for further identification of separated substances on the surface of the plate. Separation of mixture components improved with the increase of the distance, but she was the best value that for conventional plate size was 120-150 mm. This is because the capillary forces acting on the front lines of wetting and mobile phase as we move on layer chromatographic everything feels less impact capillary forces through balancing their gravity. In carrying out chromatography to obtain optimal length Front promotion period ranged from 45 minutes to 2.5 h. depending on the viscosity of the eluent. In the case of high-performance thin-layer chromatography, where wrapping layer chromatographic particles denser optimum distance was about 60 mm, and elution took 15 to 50 minutes. Then it was found that Rf1 is 0.40; Rf2 is 0.43; Rf3 is 0.48; Rf4 is 0.51; Rf5 equal to 0.53, where the value of Rf is the ratio of the path traveled by the center of the zone investigated component of the starting line to the path traversed eluent, reflecting the position of chromatographic zones in the chromatogram. As a result of 20 research facilities dried menstrual blood we found that 98% (18 cases) in the chromatogram formed purple spots absorption of fractions of takeoff from 41mm to 53 mm, which corresponded to standard indicators prostaglandin F2alfa within 40-55mm , whose presence in the investigated Properties we considered the main criterion menstrual blood.

As a result of studies, we found that the division in TLC influenced by several factors - the composition and properties of the eluent, nature, dispersion and sorbent porosity, temperature, humidity, size and thickness of the sorbent, the size of the camera. Therefore, to obtain reproducible results we were thoroughly soaked standardized research conditions. Reliability of information also confirmed relative values ​​Rs (Rf investigational compounds related to standard connection Rf), which in our study coincide with the literature data.

Conclusions: Thus, summarizing the results of the studies relating to the establishment of the regional origin of blood, including menstrual, it should be noted that in addressing this issue are important research chemical regulation of the menstrual cycle hormonal and menstrual blood composition origin, including prostaglandin F2alfa. It should be noted that the detection of a test object (dried on gauze menstrual blood) prostaglandin F2a with factions of takeoff from 41mm to 53 mm, which are in line with those of the standard prostaglandin F2alfa within 40-55mm, we can conclude that this object ' Object is menstrual blood and the presence of prostaglandin F2a the chromatogram is reliable sign of menstrual blood origin.

Literature:

1.     Abel M.N. The effect of 17 beta-estradiol and progesterone on prostaglandin production by human endometrium maintained in organ culture / M.N.  Abel // Endocrinology. - 1980. - V. 106. – Р. 1599-1606.

2.     Schatz F. Ea. Prostaglandin output by human endometrium under superfusion and organ culture conditions /  F. Ea. Schatz // Steroid Biochem. - 1985. - V. 22. – P. 231-235.

3.     Wilson T. The effect of progesterone on the release of arachidonic acid from human endometrial cells stimulated by histamine / T. Wilson // Prostaglandins - 1986. -  V. 31. – Р.  343-360.

4.     Бобылев Р.В.Технология лекарственных форм: Учебник в 2 томах / Р.В. Бобылев, Г.П. Грядунова, Л.А.Иванова [и др.]; под ред. Л.А. Ивановой. – М.: Медицина, 1980. – Т.2. – С. 64-93.

5.     Загрядская А.П. Современное состояние и перспективы развития судебно-медицинских цитологических исследований / А.П. Загрядская // Судебно-медицинская экспертиза.- 1985.-№ 2.-С.40-41.

6.     Загрядская А.П. Состояние и задачи дальнейшего совершенствования методического уровня судебно-цитологических исследований вещественных доказательств / А.П. Загрядская // Судебно-медицинская экспертиза.- 1980.-№ 1.-С. 8-10.