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German Science Herald № 3/2016

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Deutscher Wissenschaftsherold • German Science Herald, N 3/2016

REDAKTIONSKOLLEGIUM / INTERNATIONAL EDITORIAL BOARD:

Jurga Bernatoniene, Dr., Prof. Physics Lithuania [email protected] Arvaidas Galdikas, Dr. habil., professor Physics Lithuania, [email protected] Kristina Ramanauskienė, Ph.dr., Prof. Pharmacy, Lithuania [email protected] Khpaliuk Alexander, Dr. med. habil., Prof. Pharmakologie, Belorus [email protected] Arnold M. Gegechkori, Dr., full Prof. Biology, Georgia [email protected] Omari Mukbaniani, Prof., DSc. Chemistry, Georgia [email protected] Teimuraz Lezhava, Prof. Genetics, Georgia [email protected] Shota A. Samsoniya, Prof. Chemistry, Georgia [email protected] Mdzinarashvili Tamaz, DSc., Prof. Biophysics, Georgia [email protected] Aliaksandr V.Prokharau, MD, PhD, MSc Prof. Oncology, Belarus [email protected] Pyrochkin V., MD, PhD, MSc Prof. Theraphy, Belarus [email protected] Golubev A.P., BD, Prof. Ecology, Belarus [email protected] Makarevich A., MD, PhD, Prof. Theraphy, Belarus [email protected] Kanunnincova N., BD, Prof. Physiology, Belarus [email protected] Giedrius Vanagas, Prof. Internal Medicine, Lithuania [email protected] Armuntas Baginskas, Prof. Neurofiziologija, Lithuania [email protected] Ricardas Radisauskas, MD., Ph.D., Prof. Cardiology, Lithuania [email protected]

Edmundas Kadusevicius, MD, PharmD, PhD, Prof. Pharmacology, Lithuania [email protected] Ivo Grabchev, Prof., PhD. Chemistry, Bulgaria [email protected] [email protected] Mariyana Ivanova Lyubenova, Prof., PhD. Ecology, Bulgaria [email protected] [email protected] Tsvetanka Tsankova Marinova, MD, PhD, DMedSci, Biology, Bulgaria [email protected] Evgueni D. Ananiev, Prof PhD, Biology. Bulgaria [email protected] Plamen G. Mitov, Prof., PhD. Biology, Bulgaria [email protected] Atanas Dimov Arnaudov, Ph.D. Physiology, Bulgaria [email protected] Iliana Georgieva Velcheva, PhD, Ecology, Bulgaria [email protected] Osman Demirhan, Prof. Biology, Turkey [email protected] Jharna Ray, M. Sc., PhD, Prof. Neurogenetics, India [email protected] Marián Halás doc. RNDr, Ph.D. Human geography, Сzech [email protected] Ayfer Pazarbasi Prof.Dr. Biology, Turkey [email protected] Tusharkanti Ghosh Prof. Physiology, India [email protected] Khudaverdi Gambarov Gambarov, Prof. Microbiology, Azerbaijan [email protected] Rovshan Ibrahimkhalil Khalilov, Prof. Biophysics, Azerbaijan [email protected] Svitlana Antonyuk, Dr.phil., Stony Brook University, USA Linguistics Samuel M.Johnson, Prof.Dr.phil., Wells, Maine, USA Theology [email protected]

Lists of references are given according to the author’s country standards



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Dear subscribers! This issue of our magazine is devoted to cooperation of our editorial board with

Ukrainian medical scientists. We present you the results of their researches.

UDC 611.835.8:618.29

Khmara T.V., Komar T.V.

Higher State Educational Institution of Ukraine “Bukovinian State Medical University”, N.H. Turkevych Department of human anatomy, Chernivtsi, Ukraine, [email protected]

INDIVIDUAL ANATOMICAL VARIABILITY IN THE INNERVATION OF THIGH AND CALF MUSCLES IN HUMAN FETUSES

Abstract. The study of features of the topography of the sciatic nerve and its branches has been conducted on 7 human fetuses aged 6 months. We found a high division of the sciatic nerve into the tibial and common peroneal nerves in two fetuses. We described the involvement of the sciatic nerve in the innervation of the quadrate muscle of thigh, piriform and gluteus maximus muscles, as well as the part of the tibial nerve in the innervation of the long head of the biceps femoris, semitendinosus and semimembranosus muscles. We established the individual anatomical variability in the distribution of intramuscular nerves in the posterior group of thigh muscles, back, lateral and anterior groups of calf muscles. Key words: sciatic nervе, tibial nerve, superficial peroneal nerve, deep peroneal nerve, fetus, human.

Introduction. The symptoms of compression of the sciatic nerve in the infrapiriform foramen develop on the basis of its topographical and anatomical relationship with the surrounding structures. Severe lesions of the sciatic nerve are usually accompanied by severe paresis or paralysis of the back lower limb muscles [1-2]. Establishing accurate data about the features of the topography of the sciatic nerve and its branches, as well as the awareness of the general laws of the intramuscular nerve distribution in each muscle of the thigh and calf in different periods of human ontogenesis is necessary to carry out diagnostic procedures and surgical intervention [3].

Objective: to establish the features of the intramuscular distribution of nerves in the muscles of the thigh and calf in fetuses aged 6 months.

Materials and methods. The study of topographic and anatomical features of innervation of the lower limb muscles was performed on 7 human fetuses with 186,0-230,0 mm of crown-rump length (CRL) using the

methods of fine preparation, vascular injection and morphometry.

Results and discussion. While studying the features of the topography and formation of the terminal branches of the sciatic nerve in six-month-old fetuses it was established that the right sciatic nerve was divided into the tibial and common peroneal nerves on the level of the back lower third of the thigh (42.85% of cases) or the upper corner of the popliteal fossa (42.85 %); one case (14.3%) showed a high division of the sciatic nerve into its terminal branches. We found the following levels while studying the bifurcation of the left sciatic nerve: between the middle and lower third of the back of the thigh (28.6%), on the rear surface of the lower third of the thigh (42.8%), upper corner of the popliteal fossa (14.3%), a high division of the sciatic nerve was found in one fetus (14.3%).

We found high branching of the left sciatic nerve trunk into the tibial and common peroneal nerves, namely 11.0 mm below the infrapiriform foramen in the fetus with 193,0 mm of CRL. The

© Khmara T.V., Komar T.V., 2016


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extrapelvic part of the sciatic nerve is 14,0 mm long, the tibial nerve is 51,0 mm and the common peroneal nerve is 38.0 mm. The sciatic nerve gives muscular branches that innervate the piriform muscle, inferomedian part of the gluteus maximus, the quadrate muscle of thigh, the proximal part of the semitendinosus and semimembranosus muscles, as well as the upper part of the long head of the biceps muscle of thigh. After branching from the sciatic nerve, the tibial nerve goes to the popliteal fossa along the posteromedial thigh, innervating the muscles of the posterior group of the thigh.

The right sciatic nerve of a fetus with 200.0 mm of CRL leaves the pelvic cavity through the infrapiriform foramen along with the posterior cutaneous nerve of thigh, two inferior gluteal nerves, inferior gluteal vessels and sexual neurovascular bundle. The sciatic nerve runs 5.6 mm laterally to the ischial tuberosity, lying under the gluteus maximus muscle. The right sciatic nerve in the gluteal region is 9.0 mm long. At a distance of 7.0 mm below the infrapiriform foramen the sciatic nerve branches into the tibial and common peroneal nerves. The tibial nerve is 42,0 мм long from the place where it branches from the sciatic nerve and to the popliteal fossa and its total length is 87.0 mm. The common peroneal nerve is 41.0 mm long and it is located laterally to the tibial nerve. The tibial nerve branches into 4 muscular branches that approach the long head of the biceps femoris muscle from its front surface, one branch from the common peroneal nerve goes to its short head. Most of intramuscular nerve trunks in the thickness of the long and short heads of the biceps femoris go downwards. However, none of the nerve trunks penetrates into the thickness of the neighboring head. Two muscular branches which are 4.0 mm and 4.4 mm long go from the tibial nerve to the semimembranosus muscle at an acute angle and the gate for their entrance is located along the outer edge of the upper third of the muscle (Fig. 1).

In the depth of the muscle belly of the semimembranosus muscle the direction of nerve trunks coincides with that of the main arterial trunks. Three muscular branches, whose entrance lies in the areas of the proximal and

Fig. 1. Right gluteal region of a fetus with 200,0 mm of CRL. Rear view. Gross specimen. Magn. 2,3х: 1 –

infrapiriform foramen; 2 – piriform muscle; 3 – sciatic nerve; 4 – inferior gluteal nerves; 5 – tibial nerve; 6 – common peroneal nerve; 7 – muscular

branches from the tibial nerve; 8 – a muscular branch from the common peroneal nerve; 9 – long head of the biceps femoris muscle; 10 – short head of the biceps femoris muscle; 11 – semitendinous

muscle; 12 – semimembranous muscle; 13 – gluteus maximus muscle; 14 – gluteus medius muscle.

distal parts of the muscle, go at an acute angle from the tibial nerve to the semitendinosus muscle.

The common peroneal nerve 3.5 mm above the lateral epicondyle of the femur divides into superficial and deep fibular nerves. The superficial fibular nerve passes under the long peroneal muscle, then passes to the medial surface of the short peroneal muscle and in the lower third of the leg branches into the dorsal cutaneous nerves of the foot. The entrance for two branches in the long peroneal muscle, which extend at an acute angle from the superficial tibial nerve, is located in the upper third of the muscle belly 4.0-5.0 mm below the fibular bone head (Fig. 2). Two branches (anterior and posterior ones) go into the depth of the short tibial muscle on the side of the


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respective surfaces of the upper third of the muscle belly, at the level of the pass of the proximal tendon into the muscle belly.

The deep fibular nerve pierces the thickness of the initial parts of the long peroneal muscle and the extensor digitorum longus, and runs from the lateral side of the anterior tibial vessels. It should be noted that the deep fibular nerve in the upper portion of the calf is located between the extensor digitorum longus and the anterior tibial muscle whereas in the middle and lower portions it lies between the anterior tibial muscle and long radial extensor muscle of foot innervating these muscles.

Three branches enter the upper third of the muscle belly of the extensor hallucis longus

Fig. 2. Nerves of the back part of the right lower

limb in a fetus with 200,0 mm of CRL. Gross specimen. Magn. 1,8х: 1 – tibial nerve; 2 – common

peroneal nerve; 3 superficial peroneal nerve; 4 – lateral cutaneous nerve of calf; 5 –muscular

branches from the tibial nerve; 6 – semitendinous muscle; 7 – semimembranous muscle; 8 – long head of the biceps femoris muscle; 9 – short head of the

biceps femoris muscle;10 – medial head of the gastrocnemius; 11 – long peroneal muscle.

muscle at an acute angle, dividing by a mixed type; two branches from the deep peroneal nerve penetrate into the upper and middle third of the muscle belly of the anterior tibial muscle at an acute angle. Two nerve trunks in the anterior tibial muscle go downwards: they branch by the main type. In this type the branches of the anterior tibial artery in the muscle of the same name have the transverse direction. In the initial section of the anterior tibial muscle nerve trunks are located in front of the arteries, and then they change direction, the arteries intersect with the nerves and lie in front of them. The entrance for the nerve trunk from the deep peroneal nerve into the extensor hallucis longus is placed in the upper third of its muscle belly. Then the nerve trunk in the thickness of the muscle belly of the extensor hallucis longus, in turn, divides into the anterior and posterior branches, which pass in front of the arteries and cross them. The common peroneal nerve also gives the lateral cutaneous nerve of calf, being 51.0 mm long, which branches in the skin of the lateral surface of the calf.

In the popliteal fossa the tibial nerve is located superficially and laterally to the popliteal vein, which, in turn, passes laterally and posteriorly to the popliteal artery. The tibial nerve gives four short, from 1.8 mm to 2.4 mm long, muscular branches at an acute angle to the popliteal muscle. The places of entrance for the latter ones are located at the border between the middle and the lower thirds of the muscle belly, with three branches going to the middle third of the popliteal muscle belly. At the level of the lower angle of the popliteal fossa the tibial nerve divides into two branches: the anterior and posterior ones. The anterior branch of the tibial nerve spreads in the thickness of the lateral and medial parts of the soleus muscle, and the posterior branch of the tibial nerve penetrates into the thickness of the back surface of the muscle.

The entrance for the muscular branches of the tibial nerve in the medial head of the gastrocnemius muscle is located 4.0 mm above the entrance for nerves of the lateral head. The direction of intramuscular nerve trunks does not coincide with the direction of the muscle bundles. It should be noted that the main nerve trunks divide by scatter type, while the


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branches, leaving these trunks divide by main type. It should be emphasized that there are much more intramuscular nerve connections in the lateral head of the gastrocnemius muscle than in the medial head of the muscle. From the popliteal fossa the tibial nerve goes to the cruropopliteal canal whose foramen is limited anteriorly by the popliteal muscle and posteriorly by the poorly expressed tendinous arch of the soleus muscle. In the cruropopliteal canal the tibial nerve is accompanied by the posterior tibial artery and veins. Muscular branches of the tibial nerve enter the thickness of the posterior tibial muscle from its dorsal surface in the upper third of the muscle belly. Direction of the intramuscular nerve trunks does not coincide with that of the intramuscular arteries. The tibial nerve gives two branches, going at an acute angle to the flexor pollicis longus muscle from the rear surface of the muscle belly. The entrance for the branches that go from the tibial nerve to the flexor hallucis longus muscle, is located in the upper third of the muscle belly. However, in the thickness of the muscle belly the direction of the intramuscular nerves does not coincide with that of the intramuscular arteries. Then, the tibial nerve goes between the lateral edge of the long flexor pollicis longus muscle and the medial edge of the flexor hallucis longus muscle, reaches the back surface of the medial malleolus, where it divides into two terminal branches: the medial and lateral plantar nerves. The medial plantar nerve is located in the medial plantar sulcus along with the homonymous artery supplying the flexor digitorum brevis and the hallucis longus muscles, with the exception of the muscle which brings the big toe and the lateral part of the flexor hallucis brevis muscle, which are innervated by the lateral plantar nerve. Terminal branches of the medial plantar nerve are the proper plantar digital nerves. The lateral plantar nerve, which is accompanied by the homonymous artery and innervates the muscles of the little toe of the foot, the plantar quadrate muscle and three plantar interossei muscles runs in the lateral plantar sulcus.

Conclusions: 1. The sciatic nerve and its brunches are a source of innervation of the back thigh muscle group and the muscles of the back,

front and lateral groups of the calf. Muscular branches of the sciatic nerve are involved in the innervation of the muscles of the quadrate muscle of thigh, piriform and gluteus maximus muscles. In case of a high division of the sciatic nerve into the terminal branches, the muscles of the back group of thigh are innervated by the branches of the tibial and common peroneal nerves.

2. The nerves enter the thickness of the majority of the muscles of thigh and calf at an acute angle to the long axis of the muscle. The entrances for the nerves are more often located in the upper third of the muscle belly, sometimes in its middle third and, as an exception (popliteal muscle), in the lower third. The entrances for the nerves may be located both on the anterior and the posterior part of the muscle belly. In some muscles (soleus, peroneal) the entrances for nerves are located on two opposite sides of the muscle. In the semitendinosus muscle, preserving merism in its structure, separateness in the innervation is expressed.

3. In the depth of thigh and lower leg muscles the nerves are distributed unevenly. We can distinguish between loose, main and mixed forms of branching in the picture of intramuscular nerve distribution. The direction of intramuscular nerves and type of their branching can be different in the same muscle.

Prospects of further research. The study of individual anatomical variability of the distribution of the nerves and the arteries in the thigh and calf muscles in fetuses of different age is of interest.

References: 1. Ахмедова Г.М. Подгрушевидная

седалищная нейропатия: клинические варианты и алгоритм терапии / Г.М. Ахмедова, Т.В. Зимакова // Практическая медицина. – 2012. – № 57. – С. 129-131.

2. Макаров А.И. Анатомия седалищного нерва и варианты ветвления / А.И. Макаров, Н.Ю. Модянов // Бюллетень северного государственного медицинского университета. – 2013. – № 1. – C. 104-105.

3. Vasilchishina A.V. Opzioni topografia entro nervo sciatico zona gluteo nel periodo perinatale di ontogenesi umana / A.V. Vasilchishina, T.V. Khmara // Italian Science Review. – 2016. – Issue 1 (34). – PP. 134-137.


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UDC: 616.688: 537. 531:615.37

Sharapova E.N. Dnipropetrovsk State Medical Academy, Department of Urology, Operative Surgery and Topographic Anatomy,

Dnipropetrovsk, [email protected]

MORPHOLOGICAL AND FUNCTIONAL CHANGES OF RATS TESTES AFTER THE

ACTION OF ELECTROMAGNETIC FIELDS OF HIGH VOLTAGE WITH LOW FREQUENCY Abstract. In this article the author established the structural and functional changes in rats testes irradiated by an electromagnetic field with high voltage and low frequency. It is determined that the electromagnetic field causes an increase in specific volume of convoluted seminiferous tubules and a reduction in stromal components of testicular tissue of animals. Also, an analysis of the processes occurring in the testis of irradiated animals, the mechanism of the immune response of the testis to the action of damaging factor was conducted. The source of immune complexes in autoimmune processes in the testis, their distribution and the response of testicular tissue to their appearance was traced. The author proved the influence of immune complexes on the testicular tissue, where they have an effect on autoimmune processes. Key words: testis, convoluted seminiferous tubule, electromagnetic field, immunity.

Introduction. Individual variability and the complexity of the human organism in the conditions of aggressive external environment directly depends on the influence of harmful factors. One of them is electromagnetic field. The development of industry in Ukraine is of interest of both native and foreign researchers in order to prevent the damage of industrial electromagnetic fields on the body of people who work on the manufacture. At home, sharing household devices that emit electromagnetic waves, also have a negative impact on the human body. In particular the most vulnerable organs in humans body are cardiovascular system, neural, visual and reproductive system. One of the most significant factors of the impact of the electromagnetic field is the state of genetic apparatus and of the regenerative function and the effect of long-term factor may be manifested in subsequent generations. Clinical and experimental studies show the development of effects of electromagnetic fields – embryotoxic, gonadotoxic and teratogenic effects. Despite the differences of researches results about the influence of electromagnetic field, it can be assumed that the electromagnetic field is a bioactive factor which negatively affects the reproductive function, embryogenesis, and heredity of living organisms.

Objective: to definition of morpho-functional changes of rats testes after their exposure to electromagnetic field voltage of 750 kV and frequency 50 Hz.

Materials and methods. The study was conducted on the gonads of 30 Mature male albino rats of Wistar population. Control group of rats was in the basement of morphological building under concrete slabs. Experimental animals were exposed by power lines, radiating electromagnetic field, a voltage of 750 kV and frequency 50 Hz five days a week for 30 minutes each day. The control group consisted of 6 rats. The animals underwent euthanasia at 14, 30 and 45 days from the beginning of the experiment by overdose of ether anesthesia. Histological sections of testes were stained with hematoxylin and eosin. The specific content of convoluted seminiferous tubules and stromal component of testicular tissue with the subsequent statistical processing of the results was calculated.

Results and discussion. It is known that the wall of convoluted seminiferous tubule in the norm consists of 3 layers: the basal, muscular and fibrous. It is lined with spermatogenic epithelium inside and consists of 2 cell types - support (sustantivites and spermatogenic with varying degrees of maturity (spermatogonia, spermatocytes I, II, spermatids). Sustantivity form a microenvironment for the development of maturing sex cells, isolate their toxins and antigens, synthesize biologically active substances, which regulate the process of spermatogenesis, and phagocytose defective sex cells.

On the 14th day after irradiation by an electromagnetic field in the seminiferous tubules of the testes of animals the disruption and destruction of seminiferous epithelium,

© Sharapova E.N., 2016


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swelling of the membranes of convoluted and straight tubules, its deformation was detected. We have calculated the relative amount of convoluted seminiferous tubules of testes and it decreased to 79,11±3,66%, and the proportion of stromal component increased to 20,89±1,15% (p<0.05). The number of tubules with spermatozoon decreased, tubules devastated. At 30 days after exposure to electromagnetic field it was observed a further decrease in the volume fraction of the seminiferous tubules to 76,65±2,36% and increasing of the relative amount of stromal component to 23.35±1,51% (p<0.05). The number of tubules with spermatozoon decreased to 52.5±2,5% (p>0.05). The processes of destruction in elements of hepatolenticular barrier and spermatogenic epithelium of the testes took place. At the 45th day after irradiation the animals to the electromagnetic field an extensive area of sclerosis in the testes was observed. Desquamation of seminiferous epithelium, vacuolization and lysis of the cytoplasm of spermatozoon was determined. The increasing of the number of fibrous structures in the adventitia shell of the efferent tubules of the testis was defined. Vacuolization of the cytoplasm of the epithelium of the efferent tubules of the testis was observed. The relative volume of seminiferous tubules decreased to 76,21±1.15% and the volume fraction of stroma increased to 23,79±2,05% (p<0.05). The relative volume of tubules with spermatozoon was 40.3±2,2% (p<0.05).

Conclusion. During the experiment, after irradiation of the electromagnetic field with high voltage and low frequency the number of convoluted seminiferous tubules of testicular tissue decreased by 3.7% (p<0.05). Atrophy and degeneration of germinal epithelium, respectively, increasing of proportion of stromal component was determined. Fragments of the destroyed cells of seminiferous epithelium and sustentocites got into the bloodstream and become autoantigens, the appearance of which the body reacted to the activation of cellular and humoral immunity. Part of the antigens fall into the thymus, where they increased the proliferation of T-lymphocytes, which were sent to the paracortical area of lymph nodes and periarterial zone of lymph follicles of the spleen. In these areas there are specialized macrophages - Interdigitated cells, capable of translating corpuscular antigen in the molecular

form and fix on the surface complexes of the antibody+antigen. With the accumulation the molecular form of antigen was antigen-dependent propagation of T lymphocytes in these areas, which have gained the specialization to this type of antigen - fragments of seminiferous epithelium. Next, T-lymphocytes, with a sensitivity of spermatogenic epithelium layer, through hepatolenticular barrier got in the spermatogenic epithelium layer and intensified destructive processes in cells through enhancing the phagocytosis of spermatogenic epithelium. The result was a growing number of autoantigens, which led to a further increase in the number of T-lymphocytes. Under the action of antigens and T-helper cells were stimulated by the formation of plasma cells and the synthesis of autoantibodies. In many cases, the process included both cellular and humoral immunity, leading to autosensibilization of seminal gland tissue and enhance the autoimmune process in the testicle. Autoimmune reactions increased degenerative processes in the testes of irradiated rats.

Prospects of further research. In further studiesit is planned to determine the degree of damage of seminiferous epithelium of rat testes of immature, mature and senile age, irradiated by electromagnetic field.

References: 1. Гребняк М.П. Екопедіатрія / М.П.

Гребняк, С.А. Щудро, О.Б. Єрмаченко, С.В. Грищенко. — Дніпропетровськ: Пороги, 2011. – 300 с.

2. Сердюк А.М. Генофонд і здоров'я: іонізуюча радіація / А.М.Сердюк. – К.: Здоров’я, 2015. – 191 с.

3. Экологически безопасные кислородсодержащие окислители и их роль в защите человека от техногенных и биологических загрязнений / В.П. Стусь, А.В. Кравченко, В.С. Кублановский, А.Б. Величенко. – Дніпропетровськ: ООО «Акцент ПП», 2015. – 331 с.

4. Laboratory long term studies on animals subjected to an electric 50Hr field / H. Le Bars, H. Brugere, F. Pupin [et al.] // L'Energia Eleсtrica. – 2010. – N 7-8. – P. 343-361.

5. Lambrozo J. Contribution de l'expérimentation animale al'étude des effets des champ électriques et magnétique (50/60 Hr) / J. Lambrozo, H. Brugere // Energies-Sante. – 2014. – Vol. 8, N 3. – P. 365-378.


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UDC 616.311/.32-008.87-053.31(477.84)

Romaniuk L.B., Kravets N.Y.,

Pokryshko O.V., Borak V.P.,

Oliinyk N.М. SHEE “I. Horbachevsky Ternopil State Medical University of Ministry of Health of Ukraine”, Ternopil, Ukraine,

[email protected]

GUT MICROBIOTA IN INFANTS WITH PERINATAL PATHOLOGY OF THE CENTRAL

NERVOUS SYSTEM Abstract. Colonization of an infant’s body by microorganisms occurs as early as during its birth. Opportunistic microorganisms play an important role in the formation of the microbial landscape of the intestine. The study showed that infants with complicated course of early neonatal period had transient intestinal dysbiosis prevaled by opportunistic flora, namely fungi of the genus Candida, S. epidermidis and E. coli. Key words: infants, opportunistic flora, dysbiosis.

Introduction. Intestinal microflora is one of the factors that ensure the resistance of the infant’s body, the proper formation of its digestive system, the formation of its normal metabolism and immune status.

Formation of the intestinal microbiota starts on the first days of the infant’s life, and is determined, besides its status, by such factors as microbial environment, getting colostrum, the time of breast-feeding, health of the mother, the course of labor. The classic type of the formation of the intestinal microflora in healthy full-term baby is as follows: 1-3 days of life - time prevalance of aerobic microorganisms over bifidobacteria; starting with the 4th-5th days – predominance of bifidobacteria in microbiocenosis, which constitute more than 98% of all microorganisms sown [1,2].

The positive effect of bifidobacteria, which do not form spores, on the physical functions of the infant’s body is due to the fact that, producing acetic (60%) and milk (30-40%) acids, they create an acidic environment in the intestines and thus prevent the growth of pathogenic and gas-forming putrefactive flora. That is why the intestinal discharge of a healthy infant with an optimal level of bifidoflora in the gut have a sour smell, with no signs of putrefactive processes or gas-formation.

The joint stay of mother and child in the maternity institution influences positively the formation of microbiocenosis in all infants’ biotypes: colonization by bifidobacteria and lactoflora is accelerated from the first days of life, the correct relationship between aerobic and anaerobic flora gets established. In a delayed mode of the joint stay, the infant’s separation from mother in the first hours of life creates favorable conditions for colonization of the infant by opportunistic microorganisms [2,3].

Objective: to analyze the degree of dominance of a microorganism in the group of the intestinal microbiota based on a retrospective analysis of case histories of the children who were treated in the intensive care unit (ICU) of the Regional Perinatal Center in Ternopil. For this purpose, we used the constancy index C (%) and the incidence rate (Рі).

Materials and methods. The material for bacteriological study, taken with the consent of the parents were the infants’ intestinal discharges. They were immediately placed into a sterile tube or into a tube with transport medium and sent to the laboratory, then sowed on sterile vivifying media (blood IPA with 5% sheep erythrocytes, yolk-salt agar, medium

© Romaniuk L.B., Kravets N.Y., Pokryshko O.V., Borak V.P., Oliinyk N.М., 2016


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Endo, Saburo medium). Innoculations were incubated at the optimum temperature for 24-72 hours. The microorganisms were identified by the classification of Bergey. Statistical analysis of the results was performed using standard software package on a personal computer.

Results and discussion. The study involved 32 infants, among whom there were 15 (46.9%) girls and 17 (53.1%) boys. All children were treated in the ICU for perinatal CNS lesions of various origins, combined with varying degrees of prematurity – 29 (72, 5%), and intrauterine infections - 13 (32.5%).

43 strains of microorganisms were isolated from the experimental material of 27 children examined. There was no growth in 5 samples (15.6%). While studying the intestinal microflora we found the dominance of coccal flora (15-37,5%), fungi of the genus Candida were isolated in 13 (32.5%) infants, E. coli - 10 (25.0%). The degree of dominance of a microorganism in a grouping was determined by the frequency of its discovery (Table). For this purpose, the constancy index C was used, which is expressed in the share of the product of the number of samples containing the study type (p) multiplied by100% of the taken samples (P). Species, which were the most common (50%) were considered to be constant, those within 20-50% were secondary, those that do not occur frequently from 1 to 19%, those that are rare - less than 1%.

Table The degree of microorganism dominance in

the intestinal contents of infants

№ Name of the microorganism

Number of samples (n=32)

С (%)

1 S. epidermidis 11 34,4

2 S. saprophyticus 4 12,5

3 E.coli 10 31,3

4 E.cloacae 1 3,1

5 Candida spp. 13 40,6

6 E. aerogenes 2 6,3

As an infant’s gut gets colonized by microflora

gradually, there were no constant kinds among the isolated microorganisms, but S. epidermidis, E. coli and Candida spp., were referred to minor

species and the rest of the selected strains are recognized as those which occur frequently. There was no aerobic flora among five samples taken from infants with complicated course of early neonatal period. As the material was taken on the first days of life, it may not necessarily contain aerobic microorganisms.

Frequent discharge of fungi of the genus Candida and E. coli out of the stool, which indicates transient dysbiosis arouses some concern. Such changes normally can last up to 6 days of life, so it would be appropriate to repeat bacteriological examination at the end of the first week, provided a long-term stay in the perinatal center.

To assess the the incidence of the populations of different microorganisms in the bowels we used the incidence index Pi, which is the ratio of the number of strains of this species to the total number of strains (Figure).

Figure. Variability of the incidence index (Pi) of microorganisms in the intestinal microbiota of

infants with perinatal damage of the CNS

These values of incidence index Pi indicate a

high prevalence of Candida fungi in the intestines of infants, which may be due to the peculiarities of feeding due to immaturity (prematurity) and abnormalities in the regulatory function of the central nervous system on the one hand and to a separate stay of the mother and baby in the hospital while having such diseases on the other hand.

Conclusions. Infants with complicated course of early neonatal period have transient intestinal dysbiosis prevailed by opportunistic pathogens, namely fungi of the genus Candida, S. epidermidis and E. coli. Since all the children

0

0.1

0.2

0.3

0.4

0.5


11

were treated at the intensive care unit of the Regional Perinatal Center of Ternopil, not with their mothers, it can be argued that it affects the amount of saprophyte flora of this biotype and requires a further study in dynamics, on intestinal colonization by resident flora and learning its properties, depending on the type of feeding and on the technique of the latter, because some of the children were premature.

Prospects of further research. The species composition of the intestinal microflora will determine the future landscape of this habitat in infants and young children and therefore will affect the functioning of the digestive system and the formation of immunity in neonatal period. Therefore, the study of the properties of aerobic and facultative anaerobic pathogenic flora will help prevent the development of dysbiotic changes in the future, to diagnose

increasing the number of pre-clinically significant concentrations and transforming virulent properties of representatives of transient microflora in these areas.

References: 1. Дещекина М.Ф. Изучение формирования

микрофлоры кишечника у новорожденных детей при совмесном и раздельном пребывании с матерью. / М.Ф. Дещекина // Педиатрия. – 1990. – №1. – С. 13-18.

2. Filoche S. Oral Biofilms: Emerging Concepts in Microbial Ecology. / S.Filoche, L.Wong, C.H. Sissons // Journal of Dental Research. – 2010. – 89. – P.8-18.

3. Robinson C.J. From Structure to Functoin: the Ecology of HostAssociated Mscrobial Communities/ C.J.Robinson, B.J.M. Bohannan, V.B. Young // Microbiology and Molecular Biology Reviews. – 2010. – № 74. – P. 453-476.


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UDC: 616.36-002.2-071-0361:616.155.348

Balaniuk І.V. Higher State Educational Institution of Ukraine “Bukovinian State Medical University”, Department of infectious

diseases and epidemiology, Chernivtsi, Ukraine, [email protected]

REACTIVE RESPONSE OF NEUTROPHILIC GRANULOCYTES OF THE PERIPHERAL

BLOOD IN PATIENTS WITH CHRONIC HEPATITIS C Abstract. The study of reactive response of neutrophilic granulocytes in 31 patients with chronic hepatitis C showed that the reactive response of neutrophils increases by 61, 22%, and is confirmed by a growth of neutrophils shift index by 56.25% and the index of relative number of neutrophils and monocytes by 16.0%. The reactive response of neutrophilic granulocytes in patients with chronic hepatitis C increased too, which contributes to an increase of immunological reactivity by 30.07%. Key words. hepatitis C, neutrophilic granulocytes.

Introduction. Hepatitis C virus (HCV) establishes numerous connections with the immune system, causing a large number of diseases mainly autoimmune diseases such as mixed cryoglobulinemia, glomerulonephritis, arthritis, thyroiditis, and others. On the other hand, in the course of hepatitis C there is a large number of autoantibodies in the blood circulation. In our opinion, it is due to the fact that, in addition to hepatocytes, blood mononuclear cells (monocytes, macrophages), B-lymphocytes and polymorphonuclear leukocytosis are susceptible to HCV too. Infection of these immunocompetent cells determines numerous immunological disorders, occurring in more than half of patients with chronic hepatitis C [1, 2].

Objective: to establish a reactive response of neutrophilic polymorphonuclear granulocytes of the peripheral blood in patients with chronic hepatitis C.

Materials and methods. During 2014-2015 at the infectious department of Chernivtsi Regional Hospital we conducted the clinical and laboratory examination of 31 patients with chronic hepatitis C involving paraclinical methods. Patients of both sexes (21 male patients and 10 female ones), aged 26-63 years, who were to be treated in hospital were included in the experiment. The average age of the patients was 44,55 ± 1.14 years. Among them, 15 (48.39%) were older than the average

age, and 16 (51.61%) younger than the average age. The control group consisted of 30 healthy individuals (21 (70%) male patients and 9 women), aged 46,81 ± 2.41years. All the patients were taken the whole peripheral venous blood that was mixed in a clean sterile tube with an anticoagulant EDTA-K2 × 2H2O. The counting of the absolute and relative number of major populations of immune cells was carried out according to the recommendations set out in the "User's Guide" on the operation hematology analyzer [3, 4].

The statistical analysis was performed on the computer IBM Pentium-IV, the package Microsoft Excel Professional for Windows XP and the software Stat Plus Professional 2009.

Results and discussion. If there was an insufficiency we had a negative pressure and the value with "+" indicated an increased function of reactivity of neutrophilic granulocytes. The value of results that was within 1-33%, corresponded to the first degree of disorders in reactive response of neutrophilic granulocytes (NG), 34-66,7% to the II degree, more than 66,7% - III degree of disorders in reactive response of NG [5, 6].

To establish the reactive response of NG the values of absolute and relative number of major populations of immune cells in peripheral blood of patients with chronic hepatitis C (CHC) are required.

Patients with chronic hepatitis C had reduced

© Balaniuk І.V., 2016


13

Table 1 The absolute and relative number of major populations of immune cells in peripheral blood of

patients with chronic hepatitis C

№ The population of immune cells

Units of measure-

ment

Patients with chronic hepatitis

С (n=31) M±m

Virtually healthy people (n=30)

M±m

The degree of immune disorders

Р

1 agranulocytes % 29,91±0,31 32,36±1,18 -I <0,05

х.109/l 1,23±0,07 1,58±0,17 -I >0,05

2 Lymphocytes % 26,35±0,25 27,50±0,91 -I >0,05

х.109/l 1,08±0,03 1,34±0,17 -I >0,05

3 Monocytes % 3,55±0,05 4,86±0,45 -I <0,05

х.109/l 0,15±0,01 0,24±0,02 -II <0,05

4 Granulocytes % 70,09±0,73 64,86±1,81 +I <0,05

5 Neutrophilic granulocytes

% 68,19±0,67 62,63±1,61 +I <0,05

6 segmented neutrophils

% 64,45±0,61 59,53±2,89 +I >0,05

7 stab neutrophils

% 3,74±0,11 3,10±0,52 +I >0,05

8 Eosinophilic granulocytes

% 1,90±0,10 2,23±0,17 -I >0,05

9 Leucocytes х.109/l 4,11±0,27 4,88±0,97 -I >0,05

10 platelets х.109/l 168,17±10,09 202,37±2,02 -I <0,05

11 red (blood) cells

х.1012/l 4,34±0,45 4,73±1,01 -I >0,05

12 Haemoglobin (Hb)

g/l 137,58±11,17 121,75±2,17 +I >0,05

13 Increased erythrocyte sedimentation

mm/h 10,52±0,77 6,08±0,31 +III < 0,01

14 Average age of the patients

years 44,55±1,14 46,21±2,41 - > 0,05

by 8.19% number of agranulocytes due to a decrease by 36.90% of relative number of monocytes, the monocyte absolute number decreased by 60.0% and the absolute number of platelets decreased by 20.34%. At the same time the relative number of granulocytes increased

by 8.06% due to an increase of neutrophilic granulocytes by 8,88%. Such changes of absolute and relative number of immunocompetent cells confirm the presence of inflammation, as evidenced by the growth of ESR by 73.03%.


14

A significant prevalence of the absolute and relative number of granulocytes over such values as agranulocytes by 2.34 and 2.34 times (in contrast by 2.01 and 2.00 times) indicates the activation of immunocompetent cells that are involved in nonspecific immune defense of the body.

Determining the non-specific reactivity is important in competent evaluation of the immune status. Neutrophilic agranulocytes, as the largest population of immunocompetent cells, form the first line of nonspecific protection against the penetration of microorganisms into the body [7]. They contribute to the synthesis of biologically active substances (lysozyme, interferon and other cytokines) and to formation of the immune response. Based on the above, we studied the reactive response of

the peripheral blood neutrophilic granulocytes in patients with CHC [8,9].

It is shown that the index of reactive response of neutrophilic granulocytes in patients with chronic hepatitis C increases by 61.22%. Due to an increase of neutrophils shift index by 56.25%, the index of neutrophilic granulocytes and monocytes increases by 16.0%. The established index of reactive response of neutrophils indicates the compensation of endogenous intoxication, which developed in the body of patients with chronic hepatitis C, and the growth in the index of neutrophils and monocytes ratio confirms the predominance of the microphage system components over the macrophage function in the reactive response. The decrease of the absolute number of leukocytes and ESR ratio by 61,54% confirms the body’s intoxication

Table 2 Reactive response of neutrophilic granulocytes in patients with chronic hepatitis C

Immuno-hematology parameters

Units of measurement

Patients with chronic hepatitis

С (n=31) M±m

Virtually healthy people

(n=30) M±m

The degree of immune disorders

Р

Index of reactive response of the neutrophilic granulocytes

S.U. 4,24±0,31 2,63±0,25 +II <0,05

Neutrophil-lymphocyte ratio

S.U. 2,59±0,18 2,38±0,17 +I >0,05

Index of neutrophil shift

S.U. 0,058±0,006 0,032±0,002 +III <0,05

Index of leukocyte shift

S.U. 2,34±0,17 2,08±0,16 +I >0,05

Granulocyte-lymphocytic index

S.U. 3,76±0,19 4,09±0,17 -II >0,05

neutrophils and monocytes ratio

S.U. 19,21±0,27 16,56±0,42 +I <0,01

Leukocytes and ESR ratio

S.U. 0,39±0,04 0,63±0,04 -II <0,05

leukocyte index S.U. 1,66±0,10 1,47±0,07 +I >0,05

Lymphocytic index

S.U. 0,39±0,04 0,39±0,04 - >0,05

non-specific reactivity index

S.U. 40,90±0,43 43,30±069 -I <0,01

immunological reactivity index

S.U. 7,96±031 6,12±0,15 +I <0,01


15

due to the autoimmune processes, but with a predominance of infection.

An increased reactivity of neutrophilic granulocytes, involved in the formation of the immune response at all stages improves immunological reactivity of patients with chronic hepatitis C by 30.07%, which is a further positive sign of the disease.

Conclusions. 1.The reactive response of neutrophilic granulocytes increases by 61,22 % in patients with chronic hepatitis C which is confirmed by an increase of neutrophils shift index by 56.25% and the index of neutrophilic granulocytes and monocytes by 16,0 %.

2. The nonspecific immune defense in patients with chronic hepatitis C involves mononuclear and polynuclear phagocytes, but the immune response is dominated by polynuclear neutrophilic leukocytes confirming the prevalence of microphage system in the nonspecific defence of the body of patients with chronic hepatitis C.

3. Growth of the reactive response of neutrophilic granulocytes in patients with chronic hepatitis C is due to an increase of immunological reactivity by 30,07 %.

Prospects for further research. The results are the basis for establishing a general immune reactivity of patients with chronic hepatitis C.

References: 1. Гаркави Л.Х. Адаптационные реакции и

резистентность организма / Л.Х. Гаркави, Я.Б. Квакина, М.А. Уколова // Ростов н/Д: Изд. Ростовского университета, 1997. – 119 с.

2. Голубовская О.А. Вирусный гепатит – это диагноз, а не приговор / О.А. Голубовская // Зеркало недели. Украина. – 2013. – № 26. – С. 3-4.

3. Показатели крови и лейкоцитарного индекса интоксикации в оценке тяжести и определения прогноза при воспалительных, гнойных и гнойно-деструктивных заболеваниях / В.К. Островский, А.В. Маценко, Д.В. Янголенко, С.В. Макаров // Клин. лаб. диагност. – 2006. – № 6. – С. 50-53.

4. Сперанский И.И. Общий анализ крови – все ли его возможности исчерпаны? Интегральные индексы интоксикации как критерии оценки тяжести течения эндогенной интоксикации, ее осложнений и эффективности проводимого лечения / И.И. Сперанский, Г.Е. Самойленко, М.В. Лобачева // Здоровье Украины. – 2009. – № 6 (19). – С. 51-57.

5. Heim M.H., Thimme R. Innate and adaptive immune responses in HCV infections / M.H. Heim, R. Thimme // J. Hepatol. – 2014. – Vol. 61, (Suppl. 1). – P.14-25.

6. Hepatitis C virus in the new era: perspectives in epidemiology, prevention, diagnostics and predictors of response to therapy / F. Ansaldi, A. Orsi, L. Sticchi, B. Bruzzone, G. Icardi // World J. Gastroenterol. – 2014. – Vol. 7, № 20 (29). – P. 9633-9652.

7. Lavanchy D. The global burden of hepatitis C / D. Lavanchy // Liver Int. – 2009. – Suppl. 1. – P. 74-81.

8. Madaliński K. Epidemiology of HCV infection in Central and Eastern Europe / K. Madaliński, K. Zakrzewska, A. Kołakowska // Godzik Przegl Epidemiol. – 2015. – Vol. 69 (3). – P. 459-464, 581-584.

9. Occult HCV Infection: The Current State of Knowledge / M. S. Rezaee-Zavareh, R. Hadi, H. Karimi-Sari, H. M. Khosravi, R. Ajudani [et al] // Iran Red. Crescent. Med. J. – 2015. – Vol. 17 (11). – P. 174-189.


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UDC: 611.36+616-092.9+616.379-008.64

Bodnarchuk Yu.V. HSEI “Ivano-Frankivsk National Medical University”, Department of Human Anatomy, Operative Surgery and

Topographical Anatomy Ivano-Frankivsk, Ukraine, [email protected]

MORPHOLOGICAL AND FUNCTIONAL CHANGES AND CLUSTER CHARACTERISTICS

OF HEPATOCYTES IN IMMATURE RATS WITH STREPTOZOTOCIN-INDUCED DIABETES

Abstract. We have studied the cluster distribution of liver hepatocytes in immature rats and their morphological alterations on the 14th and 28th days of development and progress of diabetes mellitus (DM). The study was conducted on 3-month-old male Wistar rats. Diabetes was simulated by a single intraperitoneal administration of streptozotocin made by the firm "Sigma" (USA) at a dose of 7 mg per 100 g dissolved in 0.1 M citrate buffer (pH 4.5). It was established that in the early stages of development (14th and 28th days) of streptozotocin-induced diabetes there is a statistically significant decrease in the area of all cluster hepatocytes due to glycogenolysis, energy supply and inhibition of their protein synthesis function. Key words: morphological alteration, hepatocyte, cluster analysis, streptozotocin-induced diabetes.

Introduction. The liver is known to play a major role in both detoxification and regulation of metabolism of many substances in the human body. The liver is also involved in the synthesis of protein, vitamin A, cholesterol, and regulates glycogen synthesis from the blood glucose and, conversely, when depositing [2, 11, 12].

Diabetes mellitus (DM) is known to be a metabolic disorder and is characterized by impaired insulin secretion and action, and it results in hyperglycemia [7, 8, 11]. The latter causes disturbances in various organs, including the liver, dysfunction of which is observed in diabetic patients, especially in those patients that are characterized by poor control of blood glucose [9, 10].

The available literature does not use cluster analysis in the morphometric study of the liver.

Objective: to examine morphological and functional changes in hepatocytes of immature rats in the dynamics of a course of experimental diabetes mellitus (EDM) taking into account the results of morphometric and cluster analysis.

Materials and methods. The study was conducted on 20 three-month old male Wistar rats. The animals were divided into 2 groups: the control one (5 animals in each experimental period) and the experimental one (5 animals on the 14th and 28th days of EDM). DM simulating

was carried out by a single intraperitoneal administration of streptozotocin made by the firm "Sigma" (USA) at a dose of 7 mg per 100 g of body weight, dissolved in 0.1 M citrate buffer (pH 4.5) [4], and the control group was administered an equivalent dose of 0, 1 M citrate buffer (pH 4.5). The animals were decapitated under thiopental anesthesia and we took their blood and liver for research on the 14th and 28th days of the experiment. To confirm the development of diabetes in rats and to control hyperglycemia throughout the experiment we determined the daily blood glucose in a drop of blood taken from the caudal vein using test strips for blood glucose meter by the company "Accu-Chek" (Germany).

We used histological (hematoxylin-eosin staining) and electron microscope methods. Determination of inclusions of glycogen in hepatocytes was performed on semifine sections stained with polychrome dye Humphrey Ch.D. as modified by Eroshenko H.A. [5] and on histological sections stained by Shabadash.

Morphometry was performed on the photomaterial using the software NIH USA "Image J" in manual mode, considering the magnification. We determined the performance of the profile field area of hepatocytes and their nuclei, nuclear-cytoplasmic ratio (N/C) – the ratio of the area of the profile field of the nucleus to

© Bodnarchuk Yu.V., 2016


17

the area of the cytoplasm. Computerized data processing was performed

using statistical analysis package STATISTICA (StatSoft, Inc. (2011), STATISTICA (data analysis software system), version 10. www.statsoft.com.). We applied Student test (T-tests) for testing hypotheses about the equality of averages, whenever p <0,05, and claim the difference between averages.

Results and discussion. It should be noted that many papers deal with structural and functional changes in the liver in diabetic mature rats [6, 9, 10], and the papers that describe changes in immature animals are rare. It should be noted that some authors point out different areas of hepatocytes in different zones of the hepatic lobule in the study of pathological conditions [2, 3], which shows their morphological heterogeneity. Other researchers take into account the zonal distribution of hepatocytes in the classic liver lobule, but do not note any morphological difference [8, 10]. The majority of the findings in scientific sources are based on comparing average values of the size of hepatocytes in the hepatic lobule, without specifying the differences of morphological structure in different areas of the liver lobule [3, 6]. It should be also noted that we did not find any description of the changes of the hepatic lobule in immature rats using cluster distribution (analysis) of hepatocytes into different groups based on their morphological

characteristics. Based on the above, we decided to perform

the morphometry of mononuclear hepatocytes of the hepatic plate and conduct morphometric, histological and ultrastructural analysis of the liver hepatocytes in different zones of the hepatic lobule in immature rats. While studying the histological preparations of the liver of 3 month-old rats in the control group it was established that a polygonal shape lobule is the structural and functional unit, where we distinguish between the following zones: central (zone III) - hepatocytes of which are around the central vein, peripheral (zone I) - near the portal triad and intermediate (II zone) which is located between the peripheral and central areas. Two rows of hepatocytes form the hepatic plates that go radially from the central vein to the portal triad. It should be noted that the ultrastructural and histological features of the liver structure in the control group of immature animals are not different from the norm. [1] After analyzing the morphometric findings, all the cells were divided into 3 clusters (C). For instance, the hepatocytes C1 have the largest area of both the cells and the nuclei and small N/C (Table). Hepatocytes C2 have a medium area of cells, a large area of the nucleus and large N/C, while C3 is characterized by the smallest cell area, small nucleus and small N/C (see. Table).

In order to compare the average values of standardized characteristics between the

Table Cluster characteristic of hepatocytes of 3 month-old rats with EDM

Cluster type

Experiment duration

Hepatocyte area mcm2

Hepatocyte nuclear area mcm2

N/C

С1 Control 14 d 236,48±27,46 50,39±5,45 0,28±0,04

DM 14 d 227,58±33,10* 51,33±7,42 0,30±0,06*

Control 28 d 254,17±34,56 51,11±7,18 0,25±0,05

DM 28 d 233,91±36,55* 44,69±6,72*# 0,24±0,03*#

С2 Control 14 d 189,05±25,48 51,68±6,75 0,38±0,07

DM 14 d 159,88±27,52* 47,23±8,75* 0,42±0,06*

Control 28 d 191,19±33,73 49,60±5,99 0,35±0,08

DM 28 d 173,03±23,81* 41,03±4,91* 0,32±0,05

С3 Control 14 d 167,29±28,80 35,42±4,93 0,27±0,04

DM 14 d 151,03±25,59 33,25±5,46 0,29±0,05

Control 28 d 186,86±25,68 36,60±5,99 0,24±0,05

DM 28 d 153,53±27,03* 33,45±3,98 0,28±0,03*

Notes: 1) *compared to the control, р<0,05; 2) # compared to the previous duration of the experiment, р<0,05.


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clusters in each period of the experiment and in different areas of the hepatic lobule, we used t-Student test for independent samples. Pairwise correlation showed that clusters reliably differ (p <0.05) in the area of their hepatocytes and their nuclei. Considering the zones in which measurements of the cells were carried out, we analyzed the content of their clusters, checking the hypothesis of the links by using chi-square test of independence. Whenever p-value <0,05, we argue about the connection. Pearson's Chi-squared test (p <0,05) shows that there is a connection between the cells belonging to the certain cluster and its localization in the hepatic lobule. For instance, the hepatocytes of the control group of animals C1 are mainly localized in the peripheral zone, those with C3 - in the center, with C2 - in the intermediate zone.

After 14 days of experimental diabetes there was a statistically reliable decrease of hepatocytes area in the liver due to the breakdown of glycogen, it is especially true for C1 and C2 (see. Table). The area of hepatocyte nuclei of these clusters does not change and it increases N/C, while the morpho-metric analysis of hepatocytes C3 indicates an unreliable reduction in their area. We noticed a significant decrease in glycogen granules in periportal and intermediate zones of the liver lobule at the light optical level. Therefore, we believe that reducing the area of hepatocytes C1 and C2 is due to the activation of glycogenolysis. Our data are also confirmed by those of some authors, indicating that hepatocytes of the periportal areas perform glycogen synthesizing and glycogen accumulating functions [2, 8], while hepatocytes of the central zone (to our knowledge this is C3) carry out primarily antitoxic function.

We could observe some light and dark hepatocytes at the ultrastructural level on the 14th day of EDM. The first ones are characterized by the cytoplasm with moderate electronic-optical density and by a small number of organelles, which, in turn, have antireflective matrix, in particular, they are mitochondria and the nucleus. Dark hepatocytes have the cytoplasm with increased density, which is filled with organelles. It should be noted, that both dark and light hepatocytes can be found in one field of view of the electronic microscope. Their nuclei are mostly rounded, located in the center of

the cell, chromatin granules are evenly dispersed throughout the area of the nucleus. In some places there are some hepatocyte nuclei, where chromatin is redistributed and condenses along the inner surface of the nuclear membrane. Mitochondria are swollen, sometimes vacuolated. The number of free ribosomes and those associated with tanks of granular endoplasmic reticulum (GER) decreases somewhat. The tanks of GER are dilated, with some areas of destruction. The membranes of the Golgi complex lose their parallelism, the number of vesicles decreases. Compared to the control group of animals, biliary and vascular plasmolemma of hepatocytes have fewer microvilli.

After 28 days of EDM course the area of hepatocytes of all clusters continues reducing (see. Table). At the same time the area of the nuclei reduces as well (see Table). Reducing the sizes of the nuclei in many hepatocytes when they were exposed to other toxic compounds was observed by several authors [2], and they associated these changes with reduced functional activity of hepatocytes. There was no glycogen in specimens stained by Shabadash and in semifine sections, stained with polychrome dye, which is confirmed by the findings of electron microscopic studies. At the ultrastructural level, the most distinctive feature of EDM is "desolation" of hepatocytes or so-called "empty hepatocytes". In these hepatocytes the perinuclear cytoplasm is empty, while in the control animals we noted the accumulation of rosette-like glycogen granules in the areas around the nucleus. The light hepatocytes were in the state of granular and vacuole dystrophy here and there, and the dark ones contained apoptotic cells. Different populations of hepatocytes had: shallow invaginations of the nuclear membrane, the reduction of GER and Golgi complex, expanding perinuclear space, antireflection of mitochondria matrix and destruction of their crests.

The appearance of dark and light hepatocytes with the prevalence of the latter is also observed with other effects on the liver and may indicate the depletion of the cells themselves [6]. Some data on such cells in diabetes are also found in the papers of other researchers [7]. Our findings are consistent with the results of other researchers who noted a reduction in the area


19

of hepatocytes, especially in the peripheral area, due to the impaired depositing of glycogen by hepatocytes and its excess release [2, 7, 11, 12]. Derangements in energy supply with the change in the number and destruction and lysis of mitochondrial membranes in immature animals are also noted by other scientists [10].

Conclusions. In the early stages of streptozotocin-induced diabetes (14th and 28th days) there is a reduction in the area of hepatocytes and their nuclei, which is associated with the expressed glycogenolysis process, and disturbances in detoxification function, which was confirmed by histological and ultrastructural studies. An increase in destructively-modified mitochondria with lysis of their crests indicates a derangement in energy supply of hepatocytes. Electron-transparent nuclei, reduced number of ribosomes, the expansion of GER and perinuclear space is one of the signs of inhibition of protein synthesis hepatocyte function.

Prospects for further research. The findings of our research could be used to further study the impact of various factors and pathological action of a drug substance in the liver hepatocytes, in both the treatment of diabetes and other diseases.

References: 1. Боднарчук Ю. В. Морфологічна

характеристика часточки печінки нестатевозрілих щурів з використанням кластерного аналізу / Ю. В. Боднарчук, О. Я. Жураківська // Актуальні питання медичної науки та практики. — 2015. — Т. 2, № 82. — С. 364-372.

2. Вплив різнометалічного комплексу [Cu(dmen)2][Fe(CN)5(NO)] (dmen=N,N-диметилетилендіамін) на морфофункціональний стан печінки / Н. О. Карпезо, I. В. Бєлiнська, Т. В. Рибальченко [та ін.] // Доп. НАН України. — 2011. — № 3. С. 158-163.

3. Дєльцова О. І. Морфометричний аналіз гепатоцитів при корекції токсичного впливу пестициду 2,4-Д на печінку внутрішньоочеревинним введенням глутаргіну / О. І. Дєльцова, С. Б. Геращенко, Г. Б. Кулинич // Вісник Сумського державного університету. Серія Медицина. — 2012. — № 1. — С. 23-28.

4. Пат. № 62966. Україна, МПК 51 А 61 В 10/00. Спосіб моделювання цукрового діабету 1-го типу у тварин різного віку / В.А. Левицький, О. Я. Жураківська, В.А. Міськів, Л.М. Заяць, Р.Б. Петрів, Ю.М. Якимів, Б.М. Кіщук, Р.З. Гнатюк; заявка № u 201101566; заявл. 11.02.2011; опубл. 20.09.2011, Бюл. № 18. — 6 с.

5. Поліхромний спосіб забарвлення гістологічних препаратів / О. С. Якушко, В. І. Шепітько, Г. А. Єрошенко [та ін.] // Світ медицини та біології. — 2013. — № 3. — С. 61—64.

6. Петришен О. І. Морфологічна перебудова печінки за умов хронічної алюмінієво-свинцевої інтоксикації, іммобілізаційного стресу та гіпофункції шишкоподібної залози / О. І. Петришен, Н. О. Мельник // Буковинський медичний вісник. – 2006. – Т.10, № 4. – С. 129-131.

7. Согуйко Ю. Р. Ультраструктурні особливості печінки щура в нормі та при експериментальному цукровому діабеті на ранніх етапах його перебігу в динаміці / Ю. Р. Согуйко // Експериментальна та клінічна фізіологія і біохімія. — 2010. — № 4. — С. 12-19.

8. Early stereological changes in liver of Sprague-Dawley rats after streptozotocin injection/ A. Noorafshan, B. Esmail-Zadeh, S. Bahmanpour [et al.]/ Indian Journal of Gastroenterology. — 2005. — Vol. 24. — Р. 104-107.

9. Histological and Biochemical Effects of Azadirachta indica and Melatonin in

Streptozotocin-induced Diabetic Wistar rats / E. T. Godam, M. O. Samaila, A. O. Ibegbu [et al.] //Annals of Experimental Biology. — 2014. — Vol. 2 — № 2. — P. 9-22.

10. Lukivskaya O. Protective effect of of ursodeoxycholic acid on liver mitochondrial function in rats with alloxan- induced diabetes: Link with oxidative stress / O. Lukivskaya, E. Patsenker, V. Buko // Life Sciences. — 2007. — Vol. 80. — № 26. — P. 2394-2402.

11. Salih N. D. Histological Liver Changes in Streptozotocin induced Diabetic Mice / N. D. Salih, R. K. Muslih, S. R.Hamoodi // International Medical Journal Malaysia — 2009. — Vol. 8. — P. 10-16.

12. Streptozotocin-induced diabetes mellitus affects lysosomal enzymes in rat liver / G. B. Peres, M. A. Juliano, J. A. Aguiar [et al.] // Brazilian Journal of Medical and Biological Research. — 2014. — Vol. 47. — № 6. — С. 452-460.


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UDC: 615.275:616.61-008.64-072.74:612.465

Goroshko O.M. Higher State Educational Institution of Ukraine «Bukovinian State Medical University», Department of Pharmacy,

Chernivtsi, Ukraine Zamorskii I.I., Drachuk V.M.

Higher State Educational Institution of Ukraine «Bukovinian State Medical University», Department of Pharmacy Department of Pharmacology, Chernivtsi, Ukraine, [email protected]

INFLUENCE OF ANTIOXIDANTS ON THE MORPHOLOGICAL CHANGES IN KIDNEYS AT

EXPERIMENTAL ACUTE INJURY Abstract. Morphological changes in kidneys of white rats after the use of liposomal quercetin lipoflavon and lipin at the simulation of the myoglobinuric acute renal injury and gentamicin nephropathy have been studied. Expressed nephroprotective efficacy of lipoflavon both at one-time and repeated injection is revealed. At the same time lipin demonstrates its nephroprotective efficacy just in the case of its injection during 6 days. Key words: myoglobinuric acute renal injury, gentamicin nephropathy, liposomal quercetin lipoflavon, lipin, morphological changes, rats.

Introduction. There is a very wide range of diseases, which can be complicated by the occurrence of acute renal injury (ARI). Toxic effect of a number of substances on skeletal muscles [12] and their traumatic compression, thus developing rhabdomyolysis and myoglobinuria, is one of the leaders. Medicinal forms of ARI occur quite often due to nephrotoxic action of a number of drugs (about 60% of the cases are caused by the use of antibiotics, particularly aminoglycosides) [10].

It is well known, that renal form of ARI develops is a result of damage of the renal parenchyma, accompanied by a decrease in the intensity of tubular reabsorption, and renal hemodynamics, which occurs secondarily, leads to a drop in glomerular filtration rate [5]. According to the literature data, aminoglycoside antibiotic gentamicin, which accumulates in the cortical layer of the kidney, causes structural and functional disorders of the kidney tubules, mostly proximal, with partial preservation of functions of glomerular apparatus. Most researchers associate kidney injury with gentamicin with the formation of reactive oxygen species and the weakening of antioxidant protection [7].

It was shown that the introduction of antioxidants has a positive influence on the

development of experimental ARI [3]. This makes it possible to suggest that drugs and substances with antioxidant and membrane-stabilizing properties can be used to treat various forms of ARI [6, 11]. Particularly, it was proved experimentally, that antioxidants enhance the functional activity of the kidneys at the ARI, bring creatinine excretion to the control parameters and increase glomerular filtration rate significantly [1, 2].

Furthermore, it is still not quite clear how the improvement of renal function with antioxidants at ARI corresponds to morphological changes in the kidneys. Thus, the aim of our study is to study the morphological changes in the kidneys of rats with ARI at their correction with drugs with antioxidant action of lipoflavon and lipin.

Objective: to study of the morphological changes in the kidney tissues at the correction with lipoflavon and lipin of rhabdomyolic and gentamicin ARI.

Materials and methods. Study was performed with 24 white pubertal male outbred rats with the body weight of 120-180 g. Rhabdomyolic ARI was caused by the intramuscular injection of 50% glycerol solution at a dose of 10 mg/kg [8], gentamicin nephropathy was caused by the intramuscular injection of 80 mg/kg gentamicin

© Goroshko O.M., Zamorskii I.I., Drachuk V.M., 2016


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sulfate once a day for 6 days [9]. Lipoflavon (liposomal quercetin drug) was once injected intraperitoneally at a dose of 8 mg/kg [4] in 40 min after the injection of glycerol. Lipin, which is the basis for the lipoflavon, was injected similarly at a dose, which was equal to its content in lipoflavon (32 mg/kg) [4]. At the gentamicin nephropathy the drugs were injected similarly, but during all 6 days of the disease simulation.

Kidneys tissues for histological study were fixed in 10% solution of neutral buffered formalin for 48 hours, dehydrated in ascending alcohols, embedded in paraffin at 64 °C with following production of histological sections with thickness of 5 microns, stained with hematoxylin and eosin. For objectification and in order to improve the reproducibility of the results of quantitative research, computerized morphometry of objects in histological preparations was performed. Digital copies of optical images of areas of microscopic samples were obtained with digital camera Olympus (model C740UZ) and microscope LYUMAM-P8, bank of digital micrographs was created, and micrographs were further analyzed by the software "VideoTest - Size 5.0" (Ltd. "VideoTest", Russia).

Results and discussion. Morphological study of myoglobinuric simulated pathology confirms the displays of rhabdomyolysis, which leads not only to degenerative changes but also to coagulation necrosis in the kidneys.

Thus, in samples of animals with simulated pathology there are dilated vessels full of blood, sometimes there is perivascular hemorrhage, renal tubular epithelium is in a state of granular dystrophy (94 ± 1,0%), epithelial nuclei are not visible clearly, lumen is almost absent, and 2 ± 0,1% of epithelial cells are in state of coagulation necrosis, which is manifested as cytoplasm induration and karyopyknosis (Fig. 1, 2). There is an obstruction with myoglobin cylinders of 28 ± 0,8% of convoluted tubules of cortex and 61 ± 1,2% of excretory tubules of medulla. Myoglobin cylinders mostly expanded their lumens in their localization much.

Under the correction of ARI with lipoflavon the renal morphological state becomes better: obstruction of the lumen convoluted tubules with myoglobin cylinders is irregular and covers

Fig. 1. Kidney cortex of an intact rat. Coloration with

hematoxylin and eosin. Magnification: Objective×40, eyeglass×10.

Fig. 2. Kidney cortex of a rat at the glycerol acute

renal insufficiency on the 24th hour of the experiment

in average only 8 ± 0,5% of tubules (Figure 3). The cylinders are of various dimensions, but most of them are rather small. Degenerative processes cover 82 ± 1,3% of epithelial cells, and necrosis covers 1 ± 0,1% of them.

When using lipin the obstruction of the lumen of convoluted tubules with myoglobin cylinders is irregular and in average covers 31 ±

Fig. 3. Kidney cortex of a rat after the injection of

lipoflavon. Coloration with hematoxylin and eosin. Magnification: Objective×40, eyeglass×10.


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0,6% of tubules (Figure 4). The cylinders are small and obstruct the lumens slightly. Degenerative processes and necrotic effects are similar to ARI, particularly the dystrophy covers 93 ± 0,9 % of epithelial cells, and necrosis covers 2 ± 0,3 % of them.

In contrast to the intact animals (Fig. 5), in the kidneys of animals with gentamicin nephropathy

Fig. 4. Kidney cortex of a rat after the injection of

lipin at the glycerol acute renal insufficiency.

Fig. 5. Kidney cortex of an intact rat. Coloration with

hematoxylin and eosin. Magnification: Objective×40, eyeglass×10.

(Fig. 6) granular dystrophy of epithelial tubules (84 ± 1,4%) is observed on the 6th day of the experiment, also small areas of necrosis of tubular epithelium are observed (34 ± 0,15%). In some cases, glomeruli seem to be atrophy. At the correction with lipoflavon (Fig. 7) and lipin (Fig. 8) the dystrophy of convoluted tubule epithelium decreases to 58 ± 1,2% and 52 ± 1,6% respectively. There are no atrophied glomeruli, the signs of necrosis are observed only in isolated epithelial cells mostly after application of lipin.

Fig. 6. Kidney cortex of a rat at the gentamicin nephropathy on the 6th day of the experiment.

Coloration with hematoxylin and eosin. Magnification: Objective×40, eyeglass×10.

Fig. 7. Kidney cortex after injection of lipoflavon of a rat at the gentamicin nephropathy. Coloration with hematoxylin and eosin. Objective 40, eyeglass 10.

Fig. 8. Kidney cortex after injection of lipin of a rat

at the gentamicin nephropathy. Coloration with hematoxylin and eosin.

Objective 40, eyeglass 10.

Consequently, at the use of liposomal

quercetin – lipoflavon – Its nephroprotective efficacy is confirmed both for myoglobinuric ARI and gentamicin nephropathy, accompanied by normalization of morphological state of kidney. However, the use of lipin causes almost no morphological changes in the kidney at the


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myoglobinuric simulated pathology. At the same time, at the gentamicin nephropathy the nephroprotective efficacy of lipin is higher, perhaps because of the long-term (6 days) of the drug injection.

Conclusions. 1. According to the morphological study quercetin drug lipoflavon helps to protect kidney cells from the effects of pathogenic factors of renal damage, which is manifested in reducing of degenerative changes of nephrothelial.

2. Lipin almost does not change morphological state of kidneys in animals with acute renal injury after its one-time injection, but improves renal morphology at its long use.

Perspectives of further investigations. Obtained results give grounds to conclude that the investigated quercetin drug is effective nephroprotective agent at ARI, which allows recommending it for further clinical studies in order to implement it in clinical practice. The use of lipin may be one of the pharmacological approaches to the treatment of nephropathy.

References: 1. Горошко О. М. Лікувальні властивості

ліпофлавону і корвітину при експериментальній гострій нирковій недостатності: автореф. дис. … канд. фарм. наук / О. М. Горошко. – Х., 2009. – 20 с.

2. Горошко О. М. Нефропротекторні властивості препаратів кверцетину (корвітин та ліпофлавон) на моделі гентаміцинової нефропатії / О. М. Горошко, І. І. Заморський, О. В. Геруш // Клін. та експерим. патологія. – 2009. – Т. 8, № 3 (29). – С. 19–22.

3. Зограб’ян Р. О. Застосування антиоксидантів для лікування хронічної дисфункції ниркового алотрансплантата / Р. О. Зограб’ян // Укр. журн. нефрології та діалізу. – 2014. – № 1 (41). – С. 37–41

4. Зупанець І.А. Дослідження гострої токсичності та середньо ефективних доз кверцетину при парантеральному введенні в умовах розвитку ниркової недостатності у щурів / І.А. Зупанець, С.К. Шебеко, Д.С.

Харченко // Фармакол. та лікар. токсикологія. – 2009, № 1 (8). – С. 28–32.

5. Кондаков І. І. Вплив гліцеролу на функціонально-морфологічні показники нирок при моделюванні гострої та хронічної ниркової недостатності у щурів / І. І. Кондаков, І. І. Топчій, О. М. Кірієнко // Укр. журн. нефрології та діалізу. – 2013. – № 3 (39). – С. 14–20.

6. Новые перспективы нефропротекции / C. Ю. Штрыголь, О. В. Товчига, О. О. Койро [и др.] // Бук. мед. вісник. – 2012. – Т. 16, № 3 (63), ч. 2. – С. 35–37.

7. Balakumar P. Gentamicin-induced nephrotoxicity: Do we have a promising therapeutic approach to blunt it? / P. Balakumar, A. Rohilla, A. Thangathirupathi // Pharmacol. Res. – 2010. – Vol. 62, N 3. – P. 179–186.

8. Chander V. Reversal of experimental myoglobimtrie acute renal failure in rats by quercetin, a bioflareonaid / V. Chander, D. Singh, K. Chopra // Phamakologi. – 2004. Vol. 2773, N 1. – P. 49–56.

9. Curcumin has a palliative action on gentamicin-induced nephrotoxicity in rats / B. H. Ali, N. Al-Wabel, O. Mahmoud [et al.] // Fund. Clin. Pharmacol. – 2005. – N 4. – P. 473–477.

10. Experimental gentamicin nephrotoxicity and agents that modify it: a mini-review of recent research / B. H. Ali, M. Al Za'abi, G. Blunden [et al.] // Basic Clin. Pharmacol. Toxicol. – 2011. – Vol. 109, N 4. – P. 225–232.

11. Mitochondria-targeted antioxidant peptide SS31 prevents hypoxia/reoxygenation-induced apoptosis by down-regulating p66Shc in renal tubular epithelial cells / W. Y. Zhao, S. Han, L. Zhang [et al.] // Cell. Physiol. Biochem. – 2013. – Vol. 32, N 3. – P. 591–600.

12. Rhabdomyolysis and acute myoglobinuric renal failure in a patient with bilateral pheochromocytoma following open pyelolithotomy // I. Anaforoglu, M. E. Ertorer, F. E. Haydardedeoglu [et al.] // South Med. J. – 2008. – Vol. 101, N 4. – P. 425–427.


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UDC: 612.67:576.36:577.1

Mazur O.O. M.I. Pyrohov Vinnytsia National Medical University, Department of Anatomy and Biology, Vinnytsia, Ukraine,

[email protected]

ROLE OF ACTIVE FORMS OF OXYGEN IN THE AGING PROCESS (literature review)

Abstract. At oxidative stress the resistance of the organism is achieved by the work of many functional systems. During prolonged stress load the intersystem relations are violated due to unequal functional capabilities of various systems. This leads to disorders in the regulation of the components of functional systems and, consequently, to the emergence of pathological processes. Antioxidant defense system counteracts damaging effects of reactive oxygen to tissues. Inferiority of mobilization mechanisms of antioxidant potential provokes ability of cells to over-production of AFO, which, in general, promotes mutations, tumor growth, their invasion, damage of healthy cells and imbalance between cell proliferation and apoptosis. Key words: oxygen, oxidative stress, aging process.

It is well known that oxidation processes involving activated oxygen metabolites are essential for the existence of living organisms. They act as inter- and intracellular messengers, modulators and inductors in the regulation of biochemical and metabolic processes. They are the first and the most mobile element in the adaptive restructuring of the body under extreme conditions.

It is marked that the pathological effects occur at the excessive accumulation of AFO, peroxides and their secondary products, more specifically, at the imbalance of pro-oxidant-antioxidant system. This condition is commonly called oxidative stress. There are various factors that cause oxidative stress, and all of them as a result cause oxidative modification of macromolecules. The main of them are the excess of O2, a significant expression of inflammation with activation of neutrophils and macrophages, ionizing and ultraviolet radiation, smoking, xenobiotics, etc.

It is known that each stressful reaction is normally accompanied by a short increase in the amount of AFO. It is caused by the body adaptation to extreme conditions under which the AFO act as secondary messengers participating in signal transduction and activation of transcription factors and related genes, including those encoding antioxidant enzymes. In normal conditions AFO concentration in tissues is low

Н2О2 - 10-8 Мol, О2·- - 10-11 Мol, ОН- < 10-11 Мol.

Intensification of the processes of free radical oxidation under AFO leads to increased lipid peroxidation (LPO), oxidative modification of proteins (OMP), destruction of nucleic acids and carbohydrates, which causes structural and metabolic disorders in cells. In most cases OH-, which is able to take hydrogen atom from organic compounds with the formation of free organic radicals, is the initiator of this process:

RH + ОН → R·+ H2O. Lipid peroxidation is supposed to be one of

the main causes of damage and death of the cell as a result of AFO action. Also in this way fatty acids are oxidized that can cause violations of integrity and properties of biological membranes. The most significant biomarkers of oxidation of polyunsaturated fatty acids (UFA) are short-alkanes and alkenes, and alkanals, 2,4-alkadienals, alkatrienals, hydroxyalkenal, 4- hydroxyalkenals and their peroxides, malonic dialdehyde, normal aliphatic ketones and izoprostanes.

Lipid peroxidation is a chain reaction that is initiated by the hydroxyl radical, singlet oxygen and is catalyzed by ions of transition metals. The basic reactions of LPO are the following:

UFA–Н + R• → UFA• + RH (1) UFA• + О2 → UFA–ОО• (2)

UFA–ОО• + UFA–Н → UFA–ООН + UFA• (3) Fe2+ + UFA–ООН → Fe3+ + UFA–О• + ОН– (4) Fe3+ + UFA–ООН → Fe2+ + UFA–ОО•+ Н+ (5).

© Mazur O.O., 2016


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Thus, LPO starts with knocking out of the hydrogen atom from methylene group of unsaturated fatty acid with radicals (hydroxyl radical, radical of fatty acid or another one) (reaction 1). Produced fatty acid radical reacts with a molecule of oxygen, producing a peroxide radical (reaction 2). Peroxide radicals can take hydrogen atom from another molecules of fatty acid and to reduce to hydroperoxide (reaction 3). Produced radical reacts and there is a chain reaction that can proceed without initiating agents. The ions of iron and other transition metals are involved in spreading of chain reaction (reactions 4,5). Produced lipid peroxides are better soluble in a liquid than polyunsaturated fatty acids, thus they can be easily washed out of the membrane, which promotes self-renewal of membrane structures. Peroxides of fatty acids are unstable, and as a result of disconnection they decompose with the production of highly toxic aldehydes that damage biomolecules of the cell. In contrast to free radicals, aldehydes are stable compounds that can diffuse over large distances. The property of endogenous radicals to cause covalent modification of macromolecules and, consequently, to change the structure of biological membranes, to inhibit the activity of glycolysis and oxidative phosphorylation, the synthesis of proteins and nucleic acids is the base of their damaging effect on the cell.

LPO of endoplasmic and sarcoplasmic reticulum can cause uncontrolled outlet of Ca2 + in the cytoplasm, resulting in violated intracellular conducting of signals, changing of work of enzyme systems, etc. However, LPO is not only destructive process. LPO is significant for renewal of biological membranes, rotation of their protein and lipid components, regulation of physical and chemical properties of the membranes of cells and subcellular structures. Lipids peroxides and low molecular weight products of degradation of oxidized lipids may be involved in signal transduction, which determines the possibility of cell survival or death in stress situations.

Protein molecules are also targets for attack of AFO, which leads to changes in their secondary and tertiary structure, aggregation

and fragmentation. Due to the features of the chemical structure and the structural organization of proteins the POM is complex, which is caused by producing of a great amount of oxidized products of radical and non-radical nature. The first stage of POM is eliminating of hydrogen atom from the ά-carbon atom of the polypeptide chain with the producing of carbonyl radical, which rapidly reacts with O2 with the formation of alkylperoxyradical intermediate compound. This compound can transform into alkoxyradical, which transforms into hydroxyderivative of protein. It is believed that free radical damage of proteins has the same chain nature as the lipid oxidation. The following mechanism of protein (Pr) hydroperoxides at the action of AFO is proposed:

Pr + HO·→ Pr·+ H2O; Pr·+ O2 → PrOO·;

PrOO·+ e– → PrOO–; PrOO– +H·→ PrOOH;

PrOO + PrH → PrOOH + Pr·. The biological role of protein radicals is still

not clear, but their ability to acidify other biomolecules confirms their significant role in oxidative damage of biological systems and their oxidative effect. On the other hand, the ability of proteins to react with radical compounds is considered as a manifestation of their antioxidant properties. It was found out, that protein radicals are significant intermediaries at oxidative damage of low-density lipoproteins. It is shown that produced protein radicals can be neutralized with low molecular weight components of the antioxidant defense system, including urates and carotenoids.

The most important aspect of AFO action, due to the biological effect, is their interaction with DNA. About 100 variants of DNA damage by free radicals and modifications of pentose and the nitrogenous bases are identified. It is shown that mitochondrial DNA is damaged in 104 times more frequently than nuclear one. Obviously, this is caused by the fact that most of the AFO in the cell is formed in mitochondria.

Thus, high concentrations of AFO and lipid hydroperoxides inhibit DNA synthesis and cell division and can activate apoptosis


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(programmed cell death), which is useful for the organism, because the death of several cells can prevent progression of malignant processes and death of the whole organism. POM, caused by AFO, not just changes the amino acid residues of protein, but also violates its tertiary structure and even causes its aggregation and denaturation. As a result various functional activities of protein (enzymatic, regulatory, participation in matrix synthesis, transport of ions and lipids) decrease or disappear, and some of them promote mutations or become autoantigens.

Increasing of concentrations of AFO, due to the displacement of equilibrium between the processes of their producing and detoxification, is a precondition for violation of functional activity of cells and the development of pathological processes and aging.

Protection is carried out in two fundamentally different ways:

1) a decrease in the producing of the first AFO - by reducing of O2 content in the cell or its faster use by respiratory chain;

2) functioning of antioxidant system (AOS). Hence, the failure of antioxidant protection

due to any external influence (negative factors of the environment) intensifies the free radical oxidation. This is accompanied by changes in the conformation of lipids, which leads to disruption of structural and functional properties of biomembranes, increasing their lability and permeability, imbalance of enzyme membrane systems, disorder of electric transport chains of mitochondria, which leads to more systemic violations of the tissues and organs of the body.

Conclusions. 1. Reactions involving reactive oxygen are the most significant processes in mechanisms of self-regulation in the body.

2. The free radical oxidation (FRO) at the action of AFO is manifested in cellular metabolism both at normal state and at pathology. Intensifying of FRO processes may be the evidence of intoxication, leading to oxidative stress, which is a key part of most pathological processes.

3. With age there is hyperactivation of free radical processes in organs and tissues, that leads to irreversible damage of the membrane

structures, change of their permeability and cell death, that affects the functioning of tissues, organs and whole organism.

4. The factors that stimulate the formation of reactive oxygen are the interaction of cells with foreign materials (with viruses, infection, abnormal proteins, xenobiotics), their interaction with free radicals of fatty acids, radioactive radiation, membrane active compounds and stress.

5. At oxidative stress the resistance of the organism is achieved by the work of many functional systems. During prolonged stress load the intersystem relations are violated due to unequal functional capabilities of various systems. This leads to disorders in the regulation of the components of functional systems and, consequently, to the emergence of pathological processes

6. Antioxidant defense system counteracts damaging effects of reactive oxygen to tissues. Inferiority of mobilization mechanisms of antioxidant potential provokes ability of cells to over-production of AFO, which, in general, promotes mutations, tumor growth, their invasion, damage of healthy cells and imbalance between cell proliferation and apoptosis.

7. The state of antioxidant system depends on the course and metabolic activity of activity of endogenous antioxidants. Consistent and continuous functioning of these mechanisms enhances protective functions of the body, prolongs life and prevents aging of the organism.

8. It is established that aging is accompanied, on the one hand, by changes in production of FRO or AFO, and on the other hand - by significant changes in the AOP system, including decreased activity of its enzyme chain.

9. The use of antioxidant drugs can promote the normalization of extreme changes in the producing and utilization of active forms of oxygen, modulating the activity of antioxidant enzymes in order to maintain the adaptive capacity and detoxification properties of the organism.

10. Since FRO involving active oxygen radicals is one of the leading universal mechanisms of cell damage, the evaluation of the state of this process is recommended to be included as one


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of the methods for monitoring of health, including during treatment and preventive measures.

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UDC 616.839.2-02:[616.98:578.828

Vozna H.I.

Department of Infectious Diseases and Epidemiology of the Higher State Educational Institution of Ukraine “Bukovinian State Medical University”, Chernivtsi, Ukraine, [email protected]

THE STATE OF THE VEGETATIVE PROVISION OF CARDIAC PERFORMANCE IN

PATIENTS WITH HIV-INFECTION Abstract. Investigation of the heart rate variability (HRV) is one of the available and noninvasive method of stratification of the cardio-vascular risk, however, the rhythm and strength of cardiac contractions react to any stress influences very keenly.

A decrease of the heart rhythm variability, mainly at the expense of a vagus modulation decrease of the cardiac rhythm, to a less degree – sympathetic one, which becomes dominating, is observed in HIV-infected patients already on early stages of infection. A decrease of the functional state of the vegetative nervous system in HIV-infected patients is combined with the development of the vegetative imbalance in bicsympathicotonia. The degree of the vegetative disorders correlates with the extent of immunodepression and the level of viral loading. Such changes testify to an increased degree of the risk development of unfavorable cardiovascular events in HIV-infected individuals, for indicate to the reduction of activity of the parasympathetic part of VNS and heart “defense” from the rise of life threatening arrhythmias. Key words: HIV-infection, vegetative provision, cardiac performance.

Introduction. HIV-infected patients belong to the group of increased risk of cardio-vascular diseases, in particular, the development of asymptomatic ischemic heart disease [1.2]. This dictates the necessity to find prognostic and diagnostic methods for early verification of the cardio-vascular system involvement.

Investigation of the heart rate variability (HRV) is one of the available and noninvasive method of stratification of the cardio-vascular risk, however, the rhythm and strength of cardiac contractions react to any stress influences very keenly [3]. In this connection, cardiac rhythm change is a universal body reaction in response to different media influences what represent the result of numerous regulatory effects on cardiovascular system. The heart rhythm, at such approach, is considered to be as not only a pacemaker activity index of the sinus node, but to the greater degree, as integrated index of interaction of three factors regulating the cardiac rhythm: sympathetic, parasympathatic and humoral-metabolic [4]. Estimation of the state of the organism regulatory systems gives valuable information about a degree of their

intensity in regulating the heart rhythm, and also allows forecasting the risk of the development of pathological conditions. This provides a significant interest to practical use of the assessment methodology of VCR in HIV-infected patients.

Objective: to study the state of the vegetative support of cardiac activity in patients with HIV infection.

Materials and methods. 136 patients with HIV-infection and staying under conditions of the outpatient observation during 2013-2014 years in the regional centre of prophylaxis and combating Aids in the town of Chernivtsi have been examined. Among the indicated patients there were 66 (48.5%) men and 70 (51.5%) women aged 19-42 years. The middle age of the patients was (28.4±0.4) years.

Diagnosis of HIV-infection was made up on the basis of epidemiological anamnesis, clinical data and confirmed by detection of specific antibodies to HIV in reaction of immune-enzyme analysis and in the reaction of immunoblotting.

Among the examined patients there were 44(32.4%) patients with clinical stage I, 48(35.3%) – stage II, 23(16.9%) – stage III,

© Vozna H.I. 2016


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21(15.4%) – clinical stage IV. Clinical-epidemiological data and the results

of the laboratory methods of investigation: serological and immunological were taken into consideration making the diagnosis (including the content determination of CD4 – lymphocytes). The level of CD4 – lymphocytes was investigated after the disappearance of the symptoms of concomitant acute infectious disease (not less than in 4 weeks).

HIV-infection duration was counted from the moment of registering in a dispensary. 86(63.2%) patients were HIV-infected during not more than 5 years at the moment of the 1st inspection, 50(36.8%) patients – over 5 years.

All examined persons with clinical stages III-IV of HIV infection were receiving schemes of highly active antiretroviral therapy of the 1st line, recommended in Ukraine.

The group of comparison consisted of 30 healthy persons who, as to sex and age, were completely compared with the representatives of the groups under study.

All patients were examined according to the Order of MPH of Ukraine 55 dated 12.07.2010 “Clinical report of antiretroviral therapy of HIV-infection in adults and teenagers” [5].

The condition of the vegetative regulation of the heart rhythm was determined by means of computer cardiointervalometry using monitoring of the arterial pressure and electrocardiosygnals of daily SDM23. Investigations of the heart rhythm variability were conducted in morning hours, fasting, following 15 minutes of the patient’s staying in clinostasis. ECG was recorded during 5 minutes. HRV evaluation was carried out according to the standard repots, calculating the time and spectral parameters as to the International dimension standards, physiological interpretation of the clinical use, developed by the working group of the European society of cardiologists and Northern American society of cardiac stimulation and electrophysiology [6].

Standard complex of time and spectral indices was estimated among HRV parameters, physiological nature of which is similar, however, there are some distinctions. SDNN and TP are integral HRV indices depending upon the

activity of both parts of the vegetative nervous system (VNS) and characterizing the state of vegetative regulation in general. Change of these indices is indicative of the vegetative displacement into the preference side of one part of the VNS. Specifically, a decrease shows the involvement of all functional reserves of the organism under the influence of the central regulation of hypothalamohypophysial level, an increase – to activation of autonomous control levels situated below. LF/HF (the ratio of low-frequency and high-frequency constituents of the spectrum) also reveals the balance of the sympathetic and parasympathetic VNS parts. rMSDD, pNN50 and HF represent the influence of the parasympathetic part of VNS and the evidence of sinus arrhythmia. LF represents mainly the influence of the sympathoadrenal system on cardiac activity. VLF characterizes the influence of higher vegetative centers upon the cardiovascular subcortical center; represents the state of neurohumoral and metabolic levels of regulation [3, 6-8].

Results and discussion. Frequency of cardiac contraction in a state of rest was (74.10±0.69) per minute in all HIV-infected patients under study, what reliably exceeded indices in the group of practically healthy persons (68.5o±0.81) per minute (p<0.05), and accordingly statistically lower RRNN indices among patients in comparison with the control group. Specifically, this index in HIV-infected patients was (879.60±7.27) mc, what is reliably less of norm – (879.80±10.10) mc (p<0.050). This may be connected with a decrease of parasympathetic modulation of the cardiac rhythm and/or with the highest sympathetic tonus of the vegetative nervous system (VNS) in HIV-positive persons.

The data of our investigation indicate to a decrease of the heart rhythm variability, mainly at the expense of lowering the cardiac rhythm vagal simulation, in a less degree – sympathetic, which become the dominant one. The activity balance of the sympathetic and parasympathetic parts of the vegetative nervous system shifted to the side of the low-frequency component of the spectrum, that is, sympathicotonia is observed in the examined


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cohort of patients. The indicated changes are the most significant in HIV-positive patients with deep immunodeficiency. Thus, all HRV time parameters under investigation of patients with I-II clinical stages of HIV infection statistically considerably differed from the corresponding indices of the patients with III-IV clinical stages of illness – SDNN parameter of the patients of group I was (96.18±1.22) mc while in group II – (85.14±1.44) mc (p<0.05), rMSSD – (34.26±0.57) mc against (27.66±0.88) mc (p<0.05), pNN50 - (18.43±0.66) % and (13.36±1.01) % according to the corresponding stages (p<0.05). Comparative analysis of the spectral HRV indices in patients of the groups I-II has shown, that deepening the immunodepression degree is accompanied by more significant changes of the functional state of the vegetative nervous system at the expense of further depression force of the general spectrum of vegetative regulation (TP) – (2192,0±53.9) mc in HIV –infected persons with I-II clinical stages against (1874.3±37.4) mc in persons with III-IV clinical stages (p<0.05) and essential decrease of contribution of both low-frequency (LF) – with (773.3±22.6) mc in patients of group I till (617.6±28.8) mc in patients of group II (p<0.05), and in highly frequent fluctuations (HF) – from (504.4±22.3) mc till (367.5±25.6) mc (p< 0.05), as well as waves of a very low frequency(VLF) – from (742.5±21.8) mc in HIV-positive persons of group I to (607.1±and28.7) mc in patients of group II (p<0.05). VLF amplitude is closely connected with psychoemotional tension and functional state of the cerebral cortex. Power of VLF fluctuations of HRV is a sensitive indicator of the metabolic processes’ control and their low level reflects energy deficiency states. Constant psychoemotional tension is observed in HIV-infected patients, particularly with III-IV clinical stages, what may deepen energy-deficient state because of emaciation of the hypothalamus- hypophyseal-adrenal system.

Shifting of the sympathovagal balance to sympathicotonia is observed in the examined patients and indicative of it are statistically reliable increase of LF norm index from (58.52±0.51) n.o. at I-II clinical stages of illness to (62.14±0.59) n.o. at III-IV clinical stages

(p<0.05), HF norm decrease to (41.48±0.51) n.o. in group I of patients to (37.86±0.59) n.o. in group II (p<0.05) and, accordingly, increase of the vegetative balance index (LF/HF) from (1.45±0.06) in HIV-positive persons with I-II clinical stages to (1.68±0.09) in patients with III-IV stages of the disease (p<0.05).Taking into consideration the aforesaid, obviously, the risk of the development of the life threatening ventricular arrhythmias ( ventricular tachycardia and ventrical fibrillation) increases if there is a greater degree of immunodepression.

Reliable correlations between the parameters of HRV and the level CD4 and RRNN (r=+0.39 … + 0.44, p<0.05), CD4 and SDNN (r=+ 0.36… + 0.40, p< 0.05); CD4 and rMSSD (r=+0.38… + 0.43, p<0.05); CD4 and TP (r=+ 0.34… + 0.37, p< 0.05) confirm the role of changes of vegetative regulation in HIV-infection progress.

Investigation of the relation between viral loading and HRV parameters gave the possibility to establish statistically significant poor negative correlation with RRNN index (r= -0.31… - 0.32, p<0.05) and TP (r= -0.27…-0.29. p<.0.05).

None of the statistically reliable relation between the duration of HIV-infection and indices of the functional state of the vegetative nervous system in patients with HIV-infection was detected.

Not high values of correlation coefficient in the examined patients may be explained by a small per cent of patients– (15.4%) with the terminal stage of HIV-infection.

The data of our research agree with the researches of foreign scientists, devoted to the study and estimation of the vegetative disorders in HIV-infected patients using the analysis of HRV [7.9-12]. They also show the imbalance of the central subsegmental and autonomous segmental outlines regulation of the cardiac rhythm in HIV-infected persons. Thus, the American investigators at the head of Dominic C. Chow [7] examined 57 persons, from whom 31 persons were HIV-positive patients including 22 patients with the low level of viral loading and 26 patients made up the control group. Despite the fact that this research was limited by a small scale of selection, it showed essential decrease


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of time parameters (SDNN, rMSSD and pNN 50) in HIV-infected persons in comparison with the control group in uncorrected model. And these differences remained significant when making functional tests – at deep breathing (parasympathetic activity modulation) and orthostatic test (sympathetic activity modulation), even after correction of age and sex. No essential differences in indices of time domain among HIV-infected participants of the research with different level of virological depression were observed. Analyzing spectral indices, the investigators revealed a tendency to power lowering of high frequency waves [HF] and, at the same time, to an increase of the vegetative index (LF/HF) in HIV-infected patients in comparison with healthy persons. So, changes of HRV indices in the examined patients were connected with significant decrease of parasympathetic and simultaneous increase of sympathetic influence upon rhythm. A weak negative correlation between the parameters of time HRV analysis and the level of viral loading was observed. The same relation was found with the indices of the parasympathetic modulation of the cardiac rhythm during a trial with the fixed rate of breathing. To counterbalance, in case of active orthostatic trial, a positive relation with vegetative balance coefficient was fixed. The authors reached a conclusion concerning possible relation between the vegetative degeneration and HIV-viremia.

Investigation, carried out by Compostella C. and co-authors has shown a significant decrease of all parameters of HRV and greater HCF in HIV-positive patients in comparison with healthy individuals. It should be indicated that more considerable sympathetic-parasympathetic imbalance was observed in patients with the level CD4 – lymphocytes lower 200cells/mcl. Poor degree of correlation has been revealed between the parameters of heart rate variability and immune status of patients.

Dominic C. Chow’s investigation of the vegetative dysfunction in the patients with HIV-associated lipodystrophy [12], showed the evident disorders of the sympathetic-vagal heart rhythm modulation in the indicated cohort of patients. Thus, reliably lower indices of the heart

rate variability and apparent sympathicotonia in comparison with the group of HIV-infected individuals without the signs of lipodystrophy and healthy persons have been fixed in HIV-positive patients with lipodystrophy signs. This indicates to a considerable cardiovascular risk in such patients.

Somewhat earlier a number of Indian scientists [11] also carried out spectral analysis of ECG records and revealed a decrease of all HRV parameters including essential depression of index of the spectrum of general power of HIV seropositive individuals already on early stages without clinical manifestations of vegetative dysfunction. Such finding suggested an idea, that HRV changes might be early marker of coming sympatho-vagal imbalance and threat of the development of unfavorable cardiovascular events.

Conclusions. A decrease of the heart rhythm variability, mainly at the expense of a vagus modulation decrease of the cardiac rhythm, to a less degree – sympathetic one, which becomes dominating, is observed in HIV-infected patients already on early stages of infection. A decrease of the functional state of the vegetative nervous system in HIV-infected patients is combined with the development of the vegetative imbalance in bicsympathicotonia. The degree of the vegetative disorders correlates with the extent of immunodepression and the level of viral loading. Such changes testify to an increased degree of the risk development of unfavorable cardiovascular events in HIV-infected individuals, for indicate to the reduction of activity of the parasympathetic part of VNS and heart “defense” from the rise of life threatening arrhythmias.

Perspectives of further investigations. Subsequent investigations will consist of studying changes of indices of the functional state of the vegetative nervous system in HIV-infected patients depending upon the nozological forms of some of the most frequent attendant diseases.

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UDC 616.153:599.323.4:591.436

Nekrut D.О. M.I. Pyrohov Vinnytsia National Medical University, Department of clinical pharmacology and clinical pharmacy,

Vinnytsia, [email protected]

HYPERHOMOCYSTEINEMIA AS A FACTOR CONTRIBUTING TO THE DEVELOPMENT OF NON-ALCOHOLIC FATTY LIVER DISEASE

Abstract. Hyperhomocysteinemia is one of the pathogenetic factors of occurrence and progression of non-alcoholic fatty liver disease. Hypomethylation and activation of oxidative stress are biochemical mechanisms of hyperhomocysteinemia steatogene action. Keeping rats on a high fat diet for a long time leads to steatosis which is proved by a significant increase in cholesterol and triglycerides. Homocysteine is a factor of activation of oxidative stress in the formation of non-alcoholic fatty liver disease.This is evidenced by an increased activity of NADPH-oxidase, SOD and levels of carbonyl groups of proteins in liver homogenate in rats. Key words: physical activities of various intensity, hypercholesterolemia, endothelial dysfunction, femoral artery.

Introduction. Non-alcoholic hepatic steatosis (non-alcoholic fatty liver disease (NAFLD), fatty degeneration of liver) is the primary disease characterized by excessive accumulation of fat (mostly triglycerides) in the liver, where the proportion of hepatocytes, which histologically determine the presence of drops of fat is more than 5 % (histologically).

NAFLD covers 10-40% of the human population and therefore is considered to be one of the most common chronic diseases in the world. Due to the fact that approximately 30% of patients with steatosis develop nonalcoholic steatohepatitis (NASH), which in 10% of cases can be transformed into liver cirrhosis (LC) NAFLD is also one of the most urgent health problems. In developed countries NAFLD is registered in 20-35% of adults, and in in 75% of women who have reached the postmenopausal age [1]. The number of patients with NAFLD is growing. At present, NAFLD is considered to be one of the most frequent causes of rising transaminases in the serum of patients in Europe and the US [2, 3]. Fatty degeneration of liver is more common in individuals with obesity, type 2 diabetes, metabolic syndrome and cardiovascular diseases.

It is believed that one of the causes of NAFLD is hyperhomocysteinemia (HHC), which is a quite widespread phenomenon. For example, in

Ukraine HHC is found in 10% of healthy population and up to 43% of patients with cardiovascular disorders [4]. HHC is observed in cardiovascular diseases, in patients with renal failure, psoriasis, osteoporosis, diabetes, liver disease, miscarriages, some neuropsychiatric diseases, defects in development, carcinogenesis, etc., and there is a tendency to a permanent extension of the list [5-7]. A lot of mechanisms of pathogenic action of HHC have been described, among them the main are believed to be inhibition processes of methylation, activation of oxidative stress and protein homocysteinemia that trigger other pathological processes – destabilizing the genome (due to the decrease in DNA methylation), dysregulation of some redox-sensitive genes, reducing the synthesis of hydrogen sulphide, thrombophilia, etc. At the time there are practically no studies about the influence of HHC on the development of liver steatosis. Obviously, the research of the processes of oxidative stress, antioxidant and steatogenesis will allow to study the pro-steatogene effect of HHC.

Therefore the objective of the paper is to study chronic hyperhomocysteinemia due to the development of hepatic steatosis in rats.

Objective: to study chronic hyperhomocysteinemia due to the development

© Nekrut D.О., 2016


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of hepatic steatosis in rats. Materials and methods. The study was

conducted over 60 days at 40 nonlinear white rats weighing approximately 260 grams, which received standard starch and casein diet with saline (groups 1 and 2) or high (groups 3 and 4) fat. All the animals had free access to drinking water.

Animals of groups 1 and 2 were kept on a standard diet containing 180 g of casein, 660 g of starch and 100 g fat (50 g of lard and 50 g sunflower oil) for 1000 g of dry mixture.

The composition of the diet included a mixture of vitamins, which was prepared in accordance with physiological needs. Water-soluble vitamins were included into the the diet as their mixtures , produced on glucose. Fat-soluble vitamins were added to sunflower oil. Salt mixture and cellulose (20 g to 1000 g of dry food) were also added to the diet [8].

The animals of groups 3 and 4 were kept on a high fat diet in which 180 grams of casein accounted for 310 g of starch and 250 g fat (lard 125 g and 125 g sunflower oil).

In fact, in high fat diet the fat quota was increased up to 50 % of the total calorie content by reducing carbohydrates by 26 %.

The animals were divided into 4 groups, 10 animals in each one.

Group 1 was the control one where the rats were kept on a standard diet;

Group 2 – the rats were kept on a standard diet plus chronic HC thiolactone administering thiolactone on 1% starch solution at the rate of 100 mg / kg intragastric once a day . The control group of rats received intragastric an equivalent amount of 1% starch solution instead of thiolactone during the experiment.

Group 3 – the rats were kept on a high fat diet plus injection of 1% starch solution as in group 1of animals.

Group 4 – the rats were kept on a high fat diet plus HC thiolactone injection as in group 2 of animals.

The animals were examined daily and the amount of food eaten by them was calculated. Every week the animals were weighed.

At the end of the experiment the animals were withdrawn from the study by decapitation

under thiopental anesthesia in accordance with the European Convention for the protection of experimental animals 86/609 EEC and the Law of Ukraine № 3447-IV of 21.02.2006 "On protection of animals from cruelty." We also found biometric parameters, such as mass of the animals, body length, relative and absolute weight of the liver.

Biochemical studies. We found the activity of ALT in the serum samples (EC 2.6.1.2), AST (EC 2.6.1.1), total cholesterol (TC), triglycerides (TG), high density lipoprotein cholesterol (HDL), low density lipoprotein cholesterol (LDL), very low density lipoprotein cholesterol (VLDL). All these parameters were determined at Beskman Coulter AU analyzer 480 OLYMPUS.

To evaluate the intensity of oxidative stress, we measured the activity of NADPH-oxidase enzymes (EC 1.6.3.1), the content of malondialdehyde (MDA) and protein carbonyl groups, as well as values of antioxidant - glutathione peroxidase enzyme activity, (EC 1.11.1.9), thioredoxin reductase (EC 1.6.4.5) and reduced glutathione and superoxide dismutase (SOD) (EC 1.15.1.1) in homogenate of the liver, MDA in the homogenate and in the serum.

The level of overall HC in the serum was determined by ELISA set AXIS®.

In liver lipid extract we determined the total content of phospholipids with fero thiocyanate reagent. Malondialdehyde in the serum and liver homogenate was determined by a uniform method with thiobarbituric acid. Carbonyl groups of proteins were determined by the reaction with 2,4-dinitrophenylhydrazine. Determination of superoxide dismutase in homogenate was performed by means of quercetin by using a set of "SOD-test" production NTPK "X-Analysis" (Republic of Belarus). Reduced glutathione was determined by a standardized method with reagent of Elman. Glutathione peroxidase activity was determined by the concentration of reduced glutathione in the presence of hydrogen peroxide.

The statistical significance of differences between groups was calculated by the nonparametric method of White as the distribution of values in groups mostly did not


36

meet the normal law. The resulting values in the experimental animals are shown in the tables as M ± m, where M - arithmetic mean, m - the

average deviation from the mean. Results and discussion. It was established

(Table. 1) that during the experiment in group 4 Table 1

Values of weight in experimental animals

Values Groups of animals (explained in the text)

1 2 3 4

Rats’ weight before the experiment, g 261±15 263±19 262±20 265±32

Rats’ weight after the experiment, g 360,5±22,6 374,0±28,8 380,2±34,8 400,5±33,5*

The increase in body weight, g 99,5±15,5 111,0±18,8 118,2±21,8 135,5±21,4*

Absolute liver weight, g 9,40±0,61 11,37±1,49* 11,76±1,5* 13,83±1,27*#&

Relative liver weight,% 2,90±0,25 3,05±0,30 3,10±0,26 3,46±0,26*#&

Note: (*) - statistical significance of differences relative to group 1 exceeds 95%. (#) - Statistical significance of differences relative to group 2 exceeds 95%. (&) - Statistical significance of differences relative to group 3 exceeds 95%.

of rats which received a high fat diet in combination with the additional introduction of HC thiolactone the body weight increased reliably compared with the control group. Perhaps this is due to inhibition of thyroid function observed at HHC. Some, but not statistically reliable increase in body weight was also observed in rats of group 2. We noticed a reliable increase in absolute liver weight in all experimental groups relatively to absolute liver weight in intact animals and between the liver mass of group 4 animals relatively to absolute liver weight of animals in groups 2 and 3. However, due to the increased body weight, the relative weight of the liver only was reliably increased in the rats of group 4.

Changes of biochemical parameters (including lipid metabolism) in experimental animals are shown in Table 2. It was established that introduction of HC thiolactone to rats at a dose of 100 mg / kg for 2 months led to a pronounced HHC. For instance, HC content in the blood plasma of rats in group 2 increased by more than 1.5 times compared to the control group. It is noted that in rats of group 4 HC rate is statistically higher (almost by twice) as compared with both group 1 and group 2 and 3, which indicates the potential effect of high fat diet on HC level.

It is seen that under the influence of HHC the animals in group 2 had reliably increased levels of total cholesterol (TC), LDL cholesterol, and

atherogenic index (AI), indicating the involvementof HHC in atherogenesis. In Group 3 of the rats which received a high fat diet TG rate alone was increased.

In Group 4 rats, most values, except for HDL cholesterol levels were statistically higher than almost all other groups. Attention is drawn to the fact that the content of atherogenic LDL cholesterol and ALT were elevated as compared with group 2 (HHC) and 3 (high fat diet).

The ratio of the AST / ALT remained virtually unchanged in all groups, which can be interpreted as a lack of transition from the pathological process into chronic one.

Changes in indices of oxidative stress and antioxidant are shown in Table 3.

We found a reliable increase in MDA levels in serum and liver homogenate, NADPH-oxidase and protein carbonyl groups in liver homogenate of rats in group 2 compared to group 1. Group 3 animals that were on high fat diet increased reliably the level of all values of oxidative stress compared with group 1 except the levels of carbonyl groups of proteins. Most values of oxidative stress in rats of group 4 were higher compared with the previous groups, indicating that progression of oxidative stress in the liver tissue of animals that are kept on a high fat diet against the background of HHC.

At the same time we noted a significant reduction in the activity of antioxidant enzymes. For instance, studying antioxidant levels we


37

Table 2. Biochemical values in the serum of the experimental animals


1 2 3 4

Homocysteine, micromole/l 5,8±0,43 9,6±1,11* 7,4±1,22 11,3±0,96*#&

TCL, micromole/l 1,25±0,23 1,59±0,16* 1,35±0,22 1,76±0,28*

TG, micromole/l 0,69±0,28 0,56±0,15 0,83±0,25# 1,03±0,25*#

HDL, micromole/l 0,70±0,10 0,75±0,12 0,76±0,14 0,80±0,18

LDL, micromole/l 0,20±0,09 0,58±0,17* 0,23±0,11 0,45±0,25 *&

VLDL, micromole/l 0,32±0,13 0,26±0,08 0,33±0,11 0,46±0,10*#&

ІА, s.u. 0,77±0,19 1,20±0,38* 0,81±0,15 1,25±0,42*&

АЛТ, u/l 52,0±9,7 53,28±14,16 48,4±10,8 68,1±20,0 *#&

ACT, u/l 213,6±53,4 240,5±50,1 194,6±24,8 322,7±138,6*&

АSТ/АLТ 4,21±0,83 4,86±1,25 4,33±1,14 4,60±1,25

Note: (*) - Statistical significance of differences relative to group 1 exceeds 95%. (#) - Statistical significance of differences relative to group 2 exceeds 95%. (&) - Statistical significance of differences relative to group 3 exceeds 95%.

Table 3. Values in the system of lipid peroxidation and antioxidant defense in the liver homogenate and

serum of rats


1 2 3 4

liver homogenate

МDА, micromole/g 0,46±0,03 0,63±0,03* 0,59±0,05*# 0,91±0,08*#&

Thioredoxin reductase nmole/min*mg of protein 5,85±0,43 5.02±0.62 4,55±0,62* 4.23±0.39*

NADPH-oxidase, nmole/min on 1 mg of protein 1,25±0,14 1.47±0.09* 1,52±0,15* 1.59±0.18*

Reduced glutathione, micromole/g of protein 76,2±11,2 55,4±6,4* 68,5±3,8*# 38,4±4,3*#&

Carbonyl groups, nmole/mg of proteins 2,15±0,34 2,82±0,32* 2.45±0.22 3.25±0.33*&

Glutathione peroxidase, nmole/min*mg of protein 12,3±2,6 10,5±1,5 11,2±2,2 8,5±1,0*

SOD, u/mg of protein 1,40±0,18 1,18±0,19 1,05±0,12* 0,96±0,18*

Serum

МDА, micromole/l 3,24±0,30 4,07±0,36* 3,62±0,41 4,59±0,42*


found a reliable decrease in levels of reduced glutathione in the liver homogenate in group 2 animals compared to the control group of animals. The rats kept on high fat diet showed a significant decrease in activity of thioredoxin reductase and SOD in liver homogenate. Animals of group 4 decreased reliably (by twice) the level of reduced glutathione compared with groups 1,

2 and 3. We also noticed a reliable reduction in other values of the antioxidant protection as compared to the control group. The results indicate a decrease in functional abilities of the antioxidant system in the activation of lipid peroxidation.

Comparison of the values in hepatic steatosis is presented in Table 4.


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Table 4. Markers of hepatic steatosis in rats’ liver homogenate


1 2 3 4

liver homogenate

Phospholipids, u.opt.g. 0,176±0,008 0,146±0,020* 0,160±0,025 0,137±0,019*

Triglycerides, mmol / g 20,2±3,1 29,2±4,9* 34,6±4,2* 72,2±8,1*#&

Cholesterol, mmol / g 7,14 ±0,64 8,66±0,93* 8,31±0,82* 9,45±0,94*


The animals of groups 2 and 3 showed a

reliable increase in levels of triglycerides and cholesterol in the liver homogenate compared with animals of the control group, indicating the formation of hepatic steatosis. In group 4 of rats we noted a significant increase in all values of steatosis compared to almost all previous groups. However it should be noted, that the level of phospholipids was significantly reduced in the rats of groups 2 and 4 compared with the control group, indicating the severity of the hypomethylation process.

Conclusions. HHC is one of the pathogenetic factors of occurrence and progression of NAFLD. Hypomethylation and activation of oxidative stress are biochemical mechanisms of HHC steatogene action.

Keeping rats on a high fat diet for a long time leads to steatosis which is proved by a significant increase in cholesterol and triglycerides.

Homocysteine is a factor of activation of oxidative stress in the formation of NAFLD.This is evidenced by an increased activity of NADPH-oxidase, SOD and levels of carbonyl groups of proteins in liver homogenate in rats.

Prospects for further research. Thus, hyperhomocysteinemia accelerates the development of NAFLD significantly and knowledge of this fact will make a contribution to the development of methods to prevent accelerated development of NAFLD.

References: 1. Звенигородская Л.А. Атеросклероз и

органы пищеварения. – М.: Медпрактика, 2011.- 312 с.

2. Хворостинка В.Н. Особенности

нарушения микро- и макроэлементарного спектра сыворотки крови прижировой дистрофии печени у больных СД 2 типа /В.Н. Хвростинка, О.В. Лахно, О.И. Цивенко // Междун. эндокрин. ж. – 2007. – № 3 (9). – С. 23-26.

3. Драпкина О.М. Патогенез, лечение и эпидемиология НАЖБП – что нового? Эпидемиология НАЖБП в России. / О.М. Драпкина, В.И. Смирин, В.Т. Ивашкин // РМЖ. – 2011. – № 28. – С. 1717-1721.

4. Андрушко І.І. Рівні гомоцистеїну, цистеїну та аргініну у практично здорових осіб: вікові та статеві особливості / І.І. Андрушко // Укр. Кардіологічний журнал. – 2008. – № 5. – С. 89-95.

5. Синдром гіпергомоцистеїнемії: причини виникнення, способи профілактики та лікування /М.Б. Луцюк, Н.В. Заічко, Г.С. Григор’єва [та ін.] // Рациональная фармакотерапия. – 2013. – Т. 29, № 4. – С. 55-60.

6. Hyperhomocysteinemia and Neurologic Disorders: a Review/R.Ansari, A.Mahta, E.Mallack, J.J.Luo // J. Clin. Neurol. – 2014. – Vol. 10, № 4. – P. 281–288.

7. Faloon W. AS WE SEE IT. Newly Identified Risks Of Excess Homocysteine / W. Faloon // Life Extension Magazine. – 2015. – № 5. [Електронний ресурс http://www.lifeextension.com/Magazine/2015/5/Newly-Identified-Risks-Of-Excess-Homocysteine/Page-01].

8. Экспериментальная витаминология : Справ. руководство / Юрий Михайлович Островский . – Минск : Наука и техника, 1979 . – 552 с.


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UDC 616.6

Pelin R.H., Havrylenko O.O.

M.I. Pyrohov Vinnytsia National Medical University, Department of Radiology, Radiotherapy and Oncology, Vinnytsya, Ukraine, [email protected]

SPIRAL COMPUTED TOMOGRAPHY IN THE DIAGNOSIS OF NEPHROLITHIASIS Abstract. The analysis of SCT-examinations of 100 patients with nephrolithiasis is presented in the work. The results are generalized and indicate the grater informativeness of SCT diagnostic comparatively to other methods, and SCT diagnostic is of great practical significance for choice of the optimal treatment strategy. Key words: nephrolithiasis, calculus.

Introduction. Nephrolithiasis is the most common urological disease, associated with the formation of stones (calculuses) by crystallization of salts in urine in kidneys. According to statistics, nephrolithiasis incidence in the world ranges from 0.5% to 5.3% among the adult population and 1/3 of all diseases of the urinary tract.

For the diagnosis of nephrolithiasis, the following radiation techniques are used: ultrasound diagnostics (USD), excretory urography, spiral computed tomography (SCT). It is noteworthy that SCT can be considered as the most accurate method of radiation diagnosis of nephrolithiasis, as it is SCT that can detect almost 100% of calculuses with the size of 2 mm, allows evaluation of their structure heterogeneity. In a complex use with other methods it provides a picture of anatomical and physiological changes of pyelocaliceal system (PCS), ureters and bladder. SCT has several advantages in comparison with other radiation methods. Among the various methods of obtaining diagnostic radiographs the method of SCT without contrast is mostly used. Its application does not require special preparation of the patient and intravenous injection of contrast agents, which is contraindicated in patients with history of allergy. Informational content of this technique does not depend on the chemical composition of calculuses, which is an advantage in comparison with radiographic techniques, and on localization of calculuses, which is an advantage compared with ultrasound examination. The density of calculuses is described by the arbitrary

Hounsfield scale (HU). The density of the kidney parenchyma is from 30 to 70 HU, depending on the degree of hydration. The density of calculus is higher starting from 100 HU, but more often there are calculuses with the density varying from 500 to 1000 HU and even up to 1500 HU.

Objective: to study the SCT informativeness among patients with nephrolithiasis, to study the dependence of the nephrolithiasis incedence on the gender and age of the patients; frequency of calculuses formation in the right kidney, left kidney or both of them; localization of PCS destruction, its deformation and enlargement; number of calculuses; sizes of calculuses; calculuses shapes; state of perirenal fatty tissue; state of kidney fascia.

Materials and methods. In order to establish the nature of the pathological processes in the abdominal organs results of SCT studies in 100 patients (age, sex and numbers in %) were analyzed. The diagnostic SCT data were obtained in the clinical site research center of VNMU. SCT of the kidney were carried out using spiral computed tomography «SeleCT SP» «Elscint» (Israel), the standard method for visualization of organs of abdominal cavity and retroperitoneal space with tomographic slice thickness of 0.5 mm. Statistical analysis was performed using Microsoft Excel.

Results and discussion. The age range of 100 patients with nephrolithiasis is 23-84 years. Men suffer from nephrolithiasis more often (64 patients) than women (36 patients). The average age of male patients is – 44-62 years; the one of female patients is 45-58 years. The calculuses are found in the right kidney in 32

© Pelin R.H., Havrylenko O.O., 2016


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cases, in the left kidney - in 37 cases, and in both kidneys - in 31 cases.

Regarding PCS: calculuses are localized in the upper part in17 cases, in the middle part - in 34 cases, in the lower part - in 39 cases, and in several parts - in 10 cases. Regarding the numbers of calculuses: one calculus was found in 20 cases, less than 3 isolated calculuses - in 67 cases, numerous calculuses (more than 3) - in 13 cases. Regarding the sizes of calculuses: small ones (up to 3.0 mm) are found in 47 cases, the ones of middle size (from 3.0 to 10.0 mm) - in 30 cases, large ones (more than 10.0) - in 17 cases, numerous calculuses with various sizes - in 6 cases. Regarding the shape of calculuses: round ones are in 31 case, oval ones - in 23 cases, the ones of boulder-like shape - in 15 cases, the ones of irregular shape - in 18 cases, and numerous calculuses of various shapes - in 13 cases. The presence of PCS enlargement (hydronephrosis) is diagnosed in 12 cases, the PCS deformation is detected in 27 cases. Enhanced image of the connective tissue - bands and infiltration of perirenal fatty tissue are detected in 43 cases. Induration of renal fascias (Gerota`s fascias) is detected in 48 cases.

Conclusion. The results of nephrolithiasis studies indicate a high incidence of this disease in patients of the second mature age - 44-62 years. Nephrolithiasis is diagnosed in men more often (64%) than in women (36%). The average age of incidence is 44-62 years for men, and 45-58 years for women. The left kidney is suffered more often (37%), with isolated calculuses (67%), of small size (47%), round-shaped (31%), which are localized in the lower part of PCS (39%). PCS enlargement, which may indicate hydronephrosis, is detected in 12% of patients. Bands and infiltration of perirenal fatty tissue and Gerota`s fascias induration, which may indicate chronic inflammation, are diagnosed in 43% and 48% of cases respectively. The results are of great practical importance for choice of the optimal treatment strategy.

Prospects for further research. For further studies it is planned to take into account the anthropometric data and individual features of patients, to perform correlation analysis of accordance of constitutional features to incidence of nephrolithiasis.

References. 1. Прокоп М. Спиральная и многослойная

компьютерная томография: в 2-х т. / М. Прокоп, М. Галански ; под. ред. А.В. Зубарева, Ш.Ш. Шотемора ; пер. с англ. – 3-е изд. – М. : МЕДпресс-информ, 2007. – 710 с.

2. Хофер М. Компьютерная томография : базовое руководство / М. Хофер; пер. англ. – 3-е изд., перераб. и доп. – М.: Медицинская литература, 2011. – 232 с.

3. Петросян С. Л., Фролов М. В., Шатов А. В. Технология лучевой диагностики урогенитальных заболеваний в диагностическом госпитале // Современная диагностика в практике здравоохранения. Самара, 1995. -С.33 – 34

4. Терновой, С.К. Компьютерная томография : учебное пособие для системы послевузовского профессионального образования врачей / С.К. Терновой, А.Б. Абдураимов, И.С. Федотенков ; ред. С.К. Терновой. – М. : ГЭОТАР-Медиа, 2008. – 176 с.

5. Левковский С.Н. Мочекаменная болезнь. Прогнозирование течения и метафилактика. — СПб.: Издательство «Береста», 2010. – 136 с.

6. Рентгенологический атлас заболеваний и повреждений мочевых органов: руководство для врачей / Г.Е. Труфанов, С.Б. Петров, В.В. Рязанов и др. - СПб.: Элби-СПб, 2007. - 336 с

7. Лопаткин Н.А. Урология. – М.: Медицина, 1995. – 512 с.

8. Тіктинський О.Л. Уролітіаз. – М., 1980. – 192 с.

9. Возіанов О.Ф., Люлько О.В. Урологія.- Підручник. — 2-ге вид., доп. та перераб. — Дніпропетровськ: Дніпро-VAL, 2002. — 830 с.

10. Leder R. A., Nelson R. C. Three-dimensional CT of genitourinary tract // J. Endourol. 2001. – Vol. 15. – N 2. – P. 37 – 46.

11. Plain abdominal X-ray versus computerized tomography screening: sensitivity for stone localisation after nonenhanced spiral computerized tomography / S.V. Jackman, S.R.Potter, F.Regan [et al.] // J. Urol. – 2000. – Vol. 164.-N8.-P. 308-310.

12. Spiral computed tomography for staghorn calculi / S.N. Liberman, E.J. Halpern, K. Sullivan [et al.] // Urology. 1997. – Vol. 50. – N 10. – P. 519-524.


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UDC 351 .77:616.314–05З.2+ 577 .118

Klitynska O.V. SHEI «Uzhhorod National University», Department of Childhood Stomatology, Uzhhorod, Ukraine,

[email protected]

ANALYSIS OF THE DEPENDENCE OF STATE OF LOCAL IMMUNITY AND MICROBIAL COENOSIS IN ORAL CAVITY ON THE ACTIVITY OF CARIES IN CHILDREN RESIDING PERMANENTLY IN CONDITIONS OF BIOGEOCHEMICAL FLUORINE AND IODINE

DEFICIENCY Abstract. Aim of the study: to find out the dependence of local immunity state and microbial coenosis of oral cavity in children of different ethnic groups residing in conditions of biogeochemical fluorine and iodine deficiency on the level of activity of dental caries. Oral microbial coenosis and content of secretory immunoglobulin A in 146 children of different ethnic groups have been determined, correlations between the parameters with the level of caries activity have been revealed. When evaluating the state of local immunity in children with different levels of caries activity among the various ethnic groups it was found out that contents of secretory IgA in 100% of examined children are higher than normal; the direct correlation between the level of caries activity and sIgA concentration in the oral fluid has been revealed, moreover sIgA concentration increases with the complications of caries activity in direct proportion compared to normal healthy children (286.9 ± 6.7; 376.2 ± 7.8; 458. 2 ± 9.3; 75.5 ± 2.7; p <0.05, p <0.001). In children of the Roma ethnic group it was found out a significant increase in sIgA content in the group of children with compensated caries activity compared to the group of healthy children (135.6 ± 4.5; 59.2 ± 1.2; p <0.05), although the indicators are in the normal range (57-260 mg / ml). Key words: children, caries, oral microbial coenosis, secretory immunoglobulin A, correlation dependencies, biochemical deficiency of fluorine and iodine. .

Introduction. The study is a part of scientific theme of the department of childhood stomatology of dental faculty of SHEI "Uzhhorod National University" "Improving the dental care to children residing in conditions of biogeochemichal deficiency of fluorine and iodine" (state registration number 0114U004123), which was performed on the base of agreements on scientific and technical cooperation with the University of Presov (Slovakia, 2006), P.L. Shupyk Institute of dentistry NMAPE (2009), clinic of maxillofacial surgery of P.J. Safarik University (Slovakia, 2010), University of Debrecen (Hungary, 2010) .

In the available sources of scientific literature much attention is paid to the study of oral microbial coenosis of children and adolescents in order to establish the prevalence of certain kinds of microorganisms as etiological factor of origin and progression of major dental diseases

[7, 11-15]. However, it was found out that the main role in causing the disease is played by the local immunity and physical health of the body, especially in childhood.

The structure and antigenic properties of sIgA fundamentally differ from the ones of serum sIgA, which is a complex of three molecules of different origin and structure. The structure of sIgA includes two (or 4) sIgA molecules with a molecular weight of 160 kDa each, J-chain (16 kDa), which connects them, and secretory component that joins during the transepithelial transport of polymer sIgA. sIgA plays an important role in local immunity of the body, preventing attachment and penetration of bacteria and viruses through the epithelium, binding the pathogens that get in the body fluids and neutralizing viruses that have penetrated into epithelial cells. Several parameters of body resistance to infections correlate with the

© Klitynska O.V., 2016


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content of sIgA-antibodies in the extravascular fluids [9-12].

It was established that it is ethnic determinancy, which plays a significant role in causing the caries and its progression [1, 2, 8, 10]. Among the factors of local immunity determining by secretory immunoglobulin A [3 - 6], special attention should be paid to determining correlation dependencies of the concentration of microbial landscape and the level of activity of the dental caries in children of different ethnic groups.

Objective: to find out the dependence of local immunity and microbial coenosis of oral cavity in children of different ethnic groups residing in conditions of biogeochemical fluorine and iodine deficiency on the level of caries activity.

Materials and methods. In order to justify clinically and find out the reliability of the assumption the study of the state of oral biological coenosis and immunological state of 146 children aged 3-8 years of different ethnic groups with different levels of caries activity residing permanently in conditions of biochemical deficits was performed. The children examination was carried out after informed consent of parents or guardians in LLC "Dental clinic" of Dental Faculty of SHEI "Uzhhorod National University".

Determination of secretory IgA content was carried out by "sandwich" ELISA with the measurement of optical density value of content of plate wells with vertical scanning photometer at 450 nm.

Microbiological and immunological studies were performed in the microbiological laboratory of Transcarpathian branch of S.M. Vynogradskyi Society of Microbiologists of Ukraine (Chairman of the Society - doctor of medical sciences, professor Boiko N.V.).

Statistical analysis of the data was carried out using methods of parametric and nonparametric statistics. Consent of statistical distribution of empirical parameters with the theoretical Gaussian normal distribution was evaluated using the Kolmogorov-Smirnov (K.-S.) and Shapiro-Wilks (W) criteria, p is a reliability indicator.

At small volumes of samples the distribution was assumed to be close to normal, the arithmetic mean and the median are supposed to be nearly the same, maxima relatively to the mean values are symmetric, skewness and kurtosis do not significantly exceed 2 by their absolute value. The following methods of descriptive statistics were used: estimation of the arithmetic mean (m), confidence interval (p), median (M), minimum (min) and maximum (max), standard deviation (σ2), skewness and kurtosis. Pearson`s paired and partial correlations (r) with the confidence interval (p) were also analyzed based on the absolute data. The method of nonparametric statistics, namely the Spearman rank correlation (rs), was used too. Correlation coefficients were marked as Sk. An analysis of factor loadings, cluster analysis - hierarchical classification were performed. Allowable confidence level (p) of null statistical hypothesis (absence of significant heterogeneity or factor influence) was set equal to 0.05.

All the calculations were performed on a personal computer using licensed software "MS Excel 7" for «Windows XP» operating system.

Results and discussion. Among the 146 examined children 37 children are of the Roma ethnic group, which is 25.3%, 109 children are of other nationalities - 74.7%. The children were divided into groups due to the activity of caries: 35 ones with compensated caries (24.0%), 26 children with sub-compensated caries (17.8%), 35 children with decompensated caries (24.0%) and 50 healthy children: 25 ones of Roma ethnic group and 25 ones of other ethnic groups, which are the control group.

There are significant differences in the concentration between the children of the Roma ethnic group, the ones of mixed ethnic groups and a control group of healthy children without caries.

Immunological studies of the content of secretory IgA in the oral fluid of examined children have found out that in children with caries the sIgA content is significantly higher compared with that of healthy children (p<0.05) and higher than normal levels. In 100% of children of mixed ethnic groups contents of secretory immunoglobulin A are higher than


43

normal. Concentration of sIgA in the children of mixed

ethnic groups increased in direct proportion to the complexity of caries activity (286.9 ± 6.7; 376.2 ± 7.8; 458.2 ± 9.3; 75.5 ± 2.7; p <0.05, p <0.001). That is, there is a direct correlation between the level of caries activity in children of mixed ethnic groups and concentration of secretory immunoglobulin A in saliva, which is a marker of violations of oral microbial coenosis and indicates the further progression of lesions of dental hard tissues.

In children of the Roma ethnic group there is a significant increase of sIgA content in the group of children with compensated caries compared to the group of healthy children (135.6 ± 4.5; 59.2 ± 1.2; p <0.05), although the indicators are within normal limits ( 57 - 260 mg / ml).

When evaluating the state of local immunity in children with different levels of caries activity among the various ethnic groups it was found out that contents of secretory IgA in 100% of examined children are higher than normal; the direct correlation between the level of caries

activity and sIgA concentration in the oral fluid has been revealed, moreover sIgA concentration increases with the complications of caries activity in direct proportion compared to normal healthy children (286.9 ± 6.7; 376.2 ± 7.8; 458. 2 ± 9.3; 75.5 ± 2.7; p <0.05, p <0.001).

In children of the Roma ethnic group it was found out a significant increase in sIgA content in the group of children with compensated caries activity compared to the group of healthy children (135.6 ± 4.5; 59.2 ± 1.2; p <0.05), although the indicators are in the normal range (57-260 mg / ml).

Statistically significant correlations have been revealed between the following parameters: Streptococcus sobrinus and sIgA with r = 0.57, p = 0.01; Streptococcus sobrinus and Enterococcus faecalis and Proteus vulgaris with sIgA with r = -0.48, p = 0.07; with correlation coefficients r = (- 0.57), p = 0.01.

Table 1 shows the microorganisms having statistically significant correlative relationships one between another and with content of immunoglobulin A and the level of caries activity.

Table 1 Pearson correlation coefficients "r" and the level of reliability "p" for different pairs of

microorganisms

Microorganisms TMC Str. sobrinus

Psevdo-monas aeruginosa

Entero-coccus faecalis

Proteus vulgaris

S. aureus

TMC 1.0000 0.1004 0.4509 –0.2190 0.4482 0.7332

p= – p=0.598 p=0.012 p=0.245 p=0.013 p=0.000

Str. sobrinus 0.1004 1.0000 –0.0313 0.3673 –0.1683 –0.1974

p=0.598 p= – p=0.870 p=0.046 p=0.374 p=0.296

Psevdomonas aeruginosa

0.4509 –0.0313 1.0000 –0.0929 –0.1782 0.4562

p=0.012 p=0.870 p= – p=0.625 p=0.346 p=0.011

Proteus vulgaris 0.4482 –0.1683 –0.1782 –0.3729 1.0000 0.3852

p=0.013 p=0.374 p=0.346 p=0.042 p= – p=0.036

S. aureus 0.7332 –0.1974 0.4562 –0.2361 0.3852 1.0000

p=0.000 p=0.296 p=0.011 p=0.209 p=0.036 p= –

According to Table 1 concentration of

Streptococcus sobrinus significantly correlates with the concentration of Enterococcus faecalis in the oral fluid with the Pearson coefficient r = 0.37, p = 0.046. In its turn the concentration of Enterococcus faecalis significantly correlates

with the content of Proteus vulgaris in oral fluid with the Pearson coefficient r = -0.37, p = 0.42, though in the opposite direction.

Thus, there is an inverse dependence of the concentration of microorganisms in the oral fluid, i.e., the more the first micro-organism


44

content in the oral cavity is, the less that of the other microorganism is, in the opposite case they have antagonistic nature of the interaction in the environment.

Reliable correlations have been revealed between: concentration of Str. sobrinus and the amount of sIgA (coefficient r = 0.57, p = 0.01); concentration of Str. sobrinus and the amount of alkaline phosphatase (r = 0.40, p = 0.29); Enterococcus faecalis content and the amount of alkaline phosphatase (r = -0.49, p = 0.06), and the concentration of Enterococcus faecalis and amount of sIgA - r = -0.48, p = 0.07. The

concentration of Proteus vulgaris demonstrates counteraction to pH factors, alkaline phosphatase, Ca, P with correlation coefficient r = -0.57, p = 0.01.

As the calculation of Pearson correlation dependencies indicates no significant correlation between the factors and caries activity, the correlation dependence has been analyzed by Spearman (Sk, confidence interval was set to be p <0.05).

Table 2 shows the Spearman correlation parameters for factors that demonstrate reliable correlation with the incidence of caries.

Table 2 Spearman rank correlation

Microorganisms Streptococcus

sobrinus Enterococcus

faecalis sIgA

Streptococcus sobrinus 1.000000 0.336344 0.537879

Psevdomonas aeruginosa –0.133978 –0.101252 –0.072316

Enterococcus faecalis 0.336344 1.000000 0.446572

Lactobacillus salivarius 0.121825 0.046287 0.461528

Lactobacillus acidophilus 0.340715 –0.077074 0.096279

sIgA 0.537879 0.446572 1.000000

Caries intensity 0.575502 0.556694 0.620255 Note. * MD removed in pairs. Correlations at p <0.05 have been detected.

Thus, the Spearman correlation have been

revealed between: caries intensity and concentration of Streptococcus sobrinus (Sk = 0.575502); content of Enterococcus faecalis in oral fluid (Sk = 0.556694); content of sIgA (Sk = 0.575502); sIgA amount in its turn correlates with the content of Streptococcus sobrinus (Sk = 0.537879) and content of Enterococcus faecalis in oral fluid (Sk = 0.446572).

Amount of Streptococcus sobrinus has the relationship with the contents of Enterococcus faecalis and Lactobacillus acidophilus in oral fluid (Sk = 0.336344 - 0.340715 Sk = respectively).

Conclusions. When evaluating the state of local immunity in children with different levels of caries activity among the various ethnic groups it was found out that contents of secretory IgA in 100% of examined children are higher than normal; the direct correlation between the level of caries activity and sIgA concentration in the oral fluid has been revealed, moreover sIgA concentration increases with the complications of caries

activity in direct proportion compared to normal healthy children (286.9 ± 6.7; 376.2 ± 7.8; 458. 2 ± 9.3; 75.5 ± 2.7; p <0.05, p <0.001).

In children of the Roma ethnic group it was found out a significant increase in sIgA content in the group of children with compensated caries activity compared to the group of healthy children (135.6 ± 4.5; 59.2 ± 1.2; p <0.05), although the indicators are in the normal range (57-260 mg / ml).

An increase in the concentration of secretory immunoglobulin A is a marker of violations of oral microbial coenosis and indicates the further progression of lesions of dental hard tissues.

The Spearman correlation coefficients (Sk) has been found out between the following parameters: the caries intensity and content of Streptococcus sobrinus (Sk = 0.575502); content of Enterococcus faecalis (Sk = 0.556694); amount of sIgA (Sk = 0.575502). Concentration of sIgA in its turn correlates with amount of Streptococcus sobrinus (Sk = 0.537879) and Enterococcus faecalis (Sk = 0.446572); contents of


45

Enterococcus faecalis and Lactobacillus acidophilus have relationship with amount of Streptococcus sobrinus (Sk = 0.336344 and S k = 0.340715, respectively; confidence interval is set to be p <0.05).

References: 1. Клітинська О.В. Особливості

мікробного складу порожнини рота у дошкільнят з декомпенсованою формою множинного карієсу, які постійно проживають в умовах біогеохімічного дефіциту фтору та йоду / О.В.Клітинська // Молодий вчений. – 2015. – №5(20). – С. 27-30.

2. Клітинська О.В. Сучасні погляди на вплив окремих представників мікрофлори на розвиток стоматологічних захворювань та уражень шлунково-кишкового тракту / О.В. Клітинська, Ю.О. Мочалов, Н.В. Пупена // Молодий вчений – 2014. – №11 (14). – С. 217-220.

3. Леус П.А. Биофильм на поверхности зуба и кариес / П.А. Леус. – М.: STBOOK, 2008. – 88с.

4. Мартынова Е.А. Полость рта как локальная экологическая система / Е.А. Мартынова // Стоматология. – 2008. – №3. – С.68-75.

5. Ajami B. Evaluation of Salivary Streptococcus mutans and Dental Caries in Children with Heart Diseases / B. Ajami, G. Abolfathi, E. Mahmoudi, Z. Mohammadzadeh // J Dent Res Dent Clin Dent Prospects. – 2015. – Vol. 9(2). – P. 105-108.

6. Arora N. Microbial role in periodontitis: Have we reached the top? Some unsung bacteria other than red complex / N. Arora, N. Chugh, A. Mishra // J Indian Soc Periodontol. – 2014. – Vol. 18(1). – Р. 9-13.

7. Belstrom D. Bacterial profiles of saliva in relation to diet, lifestyle factors, and socioeconomic status / D. Belstrom, P. Holmstrup, C. H. Nielsen, N. Kirkby, S. Twetman, B L. Heitmann, V. Klepac-Ceraj, B J. Paster, N.E. Fieh // J Oral Microbiol. – 2014. – Vol. 6. – P. 236-239.

8. Klitynska O.V. Analysis of the main activity cariesgenic microbes by assessing the antibiotic activity / Klitynska O.V.// Intermedical journal. – 2015. – № 1., (Vol. III). – Р. 15-20.

9. Hajishengallis G. Role of complement in host-microbe homeostasis of the periodontium / G. Hajishengallis, T. Abea, T. Maekawa, E. Hajishengallis, J.D. Lambris // Semin Immunol. – 2013. – Vol. 25(1). – Р.65-72.

10. Hyde E.R. Metagenomic Analysis of Nitrate-Reducing Bacteria in the Oral Cavity: Implications for Nitric Oxide Homeostasis / E.R. Hyde, F. Andrade, Z. Vaksman, K. Parthasarathy, H. Jiang // PLoS ONE. – 2014. – Vol. 9(3). – Р. 11-13.

11. Langfeldt D. Composition of Microbial Oral Biofilms during Maturation in Young Healthy Adults / D. Langfeldt, S.C. Neulinger, W. Heuer, I. Staufenbiel, S. Kunzel // PLoS ONE. – 2014. – Vol. 9(2). – Р. 874 – 889.

12. Ma S. Antagonistic effect of protein extracts from Streptococcus sanguinis on pathogenic bacteria and fungi of the oral cavity / S. Ma., H. Li, C.Yan, D. Wand, H. Li, X. Xia, X. Dong, Y. Zhao, T. Sun, P. Hu, W. Guam // Experimental and therapeutic medicine. – 2014. – Vol.7. – Р. 1486-1494.

13. Mahajan A. Interspecies communication and periodontal disease /A. Mahajan, B. Singh, D. Kashyap, A. Kumar, P. Mahajan // The Scientific World Journal. – 2013. – Vol. 13. – P. 2–8.

14. Shi Y. WHO pathfinder caries survey in Beijing extended with data for prevalence of mutans streptococci /Y. Shi, D. Barmes, D. Bratthall //Brit. Dent J. – 2012. – Vol . 42. – P . 31-36.

15. Singh R.P. Salivary mutans streptococci and lactobacilli modulations in young children on consump– tion of probiotic ice–cream containing Bifidobacterium lactis Bb12 and Lactobacillus acidophilus La5 / R.P. Singh, S.G. Damle, A. Chawla // Acta Odontol Scand. – 2011. – Vol. 69 (6). – P. 389-394.


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UDC 611.846.4.013

Bambuliak A.V. Higher State Educational Institution of Ukraine “Bukovinian State Medical University”, Department of Surgical and

Pediatric Dentistry, Chernivtsi, Ukraine, [email protected]

Boichuk O.M. Higher State Educational Institution of Ukraine “Bukovinian State Medical University”, M.H. Turkevych Department of

Human Anatomy, Chernivtsi, Ukraine, [email protected]

Honcharenko V.A. Higher State Educational Institution of Ukraine “Bukovinian State Medical University”, Department of Surgical and

Pediatric Dentistry, Chernivtsi, Ukraine

TOPOGRAPHIC AND ANATOMICAL FEATURES OF NASOLACRIMAL DUCT Abstract. The study was performed on 42 specimens of nasal area of people of various ages and series of histological sections. The formation and syntopy features of the nasolacrimal duct with the adjacent structures has been studied. It has been found out that the anlage and formation of the nasolacrimal duct begins in embryo of 12.0-14.0 mm parietal-coccygeal length, the shape and topographic location of the nasal duct orifice changes with age. Keywords: nasolacrimal duct, human, ontogenesis, anatomy.

Morphological studies of nasolacrimal duct are aimed at the disclosure of mechanisms and pathogenesis of diseases and the search for effective treatments. The study of the formation and morphology of nasolacrimal duct is of great practical value, because there are variations of its structure and defects that require treatment and surgical correction [1].

According to the World Health Organization, today there are more than 160 million people with serious visual impairment in the world. In most cases the visual impairment begin in childhood and is the result of congenitally-hereditary diseases, including the ones caused by complications of the prenatal period, fetal disorders or hereditary factors [2]. Dacryocystitis of newborns (congenital malformation of lacrimal paths) is a consequence of underdevelopment of lacrimal paths or delays in reverse development of gelatinous membrane that covers the hole of nasolacrimal duct during the prenatal period. About 35% of newborns are born with this membrane and if it is not pushed at the first respiratory movements the dacryocystitis of newborns appears. Also the nasolacrimal canal patency can be affected at the presence of multiple folds, diverticula, thickening of mucosa in the inferior turbinate, lacrimal ducts

strictures, due to irregular teeth anlage, at low altitude of nasal cavity, at the face damage during the childbirth with forceps and at hereditary predisposition to the disease [3,4]. From 5 to 10% of children are born with an inability of the lacrimal paths [5]. Despite the apparent simplicity of treatment frequency of relapses and complications according to different authors varies from 12 to 26% [6,7].

Now there is a need for a detailed study of the anatomical variability in the normal range in order at the presence of individual anatomical structure features or location of the organ surgeons can be based no longer on intuition but on scientific data [4, 7, 8].

Objective: to study the formation and topographic and anatomical interrelations of nasolacrimal duct with the adjacent structures during human ontogenesis.

Materials and methods. The formation and syntopy of nasolacrimal duct has been studied on 42 specimens of corpses of people of various ages and series of histological sections. The study was performed on corpses of people who died from causes unrelated to the ENT disorders in strict accordance with the Helsinki Declaration of the World Medical Association "Ethical Principles for Medical Research Involving Human Subjects" (1964-2000). The

© Bambuliak A.V., Boichuk O.M., Honcharenko V.A., 2016


47

study of the nasolacrimal duct was carried out in Chernivtsi regional forensic medical bureau and M.H. Turkevych department of human anatomy of SHEI of Ukraine "Bukovinian State Medical University". The side walls of the nose and formations adjacent to them were dissected consistently with surgical instruments, starting from their posterior.

Results and discussion. In embryos of 8.0-10.0 mm parietal-coccygeal length (PCL) in the area of the face the lateral nasal and maxillary processes limit the nasolacrimal groove that connects the beginnings of the eye and nasal chambers. The ectoderm lining the groove is thickened and deepened in the subjacent undifferentiated mesenchyme. In embryos of 12.0-14.0 mm PCL the lateral nasal processes are partly connected to the maxillary ones, resulting the nasolacrimal groove turns into the epithelial cord. It starts from the medial corner of the eye slit, immerses in the mesenchymal cells downwards and inwards, directing to the primary nasal cavity.

At the end of the pre-fetal stage the epithelial cord, which is lined with the multilayered epithelium, elongates toward the nasal cavity and gradually turns into the hollow channel, the wall of which is already the two-layer epithelium. On the 6-7th month of embryo development the nasolacrimal duct approaches the lower nasal passage, which ends blindly. They are separated with a small layer of connective tissue. In horizontal sections the duct has a rounded shape with a diameter of 1.4 mm. Its lumen is filled with gelatin-like substance. The nasolacrimal duct is almost straight and is topographically determined slightly ahead of the upper turbinate, it crosses the front edge of the middle turbinate, and crosses the inferior turbinate at a distance of 3.0-5.0 mm from the front edge. The duct is separated from the front wall of the maxillary sinus with a layer of connective tissue of small thickness. There are connections of the duct with the nasal cavity in 6 cases.

The nasolacrimal duct of newborns has a round shape with a diameter not exceeding 2.0 mm and length of 10.5-12.8 mm. In 6 specimens it forms a small bend to the entrance to the

nasal cavity. The part of the duct that bulges mostly, is located at the front edge of the middle turbinate. In other cases the duct has a straight course, it is situated at a small distance from the front edge of the upper turbinate, crosses the front edge of the middle turbinate and crosses the inferior turbinate at a distance of 6.0-7.5 mm inwards from its front edge. The maxillary sinus is located in 0.3 mm from the back surface of the duct. The front cells of ethmoidal labyrinth are separated from it with a layer of connective tissue. In 8 study cases the lateral folds of the mucosa are detected (4 – near the mouth, 2 – In the upper third and 2 – In the transition of the lacrimal sac to the duct). It should be noted that all folds are located at its side wall. The duct orifice has slit-like shape and is elongated in the anterior-posterior direction. When the duct orifice is in the lower end of the nasolacrimal canal, it is round or oval. In 4 specimens the nasal orifice is closed with the connective tissue membrane.

While studying the lower nasal passage a small round-shaped bladder with small diameter was detected. After its cut a small amount of clear liquid appears. The revealed membranes are the anomalies of the development of the area. It may be a cause of the dacryocystitis of newborns [3]. Therefore, the ENT examination is recommended for the diagnosis of the dacryocystitis in babies at the first month of life.

During the first years of life, structure and syntopy of the nasolacrimal duct barely changes. Only the front wall of the maxillary sinus is already near the duct. At the end of children age the length of the nasolacrimal duct increases almost twice. Its lumen diameter increases by 0.2-0.3 mm. The nasal duct orifice is located under the base of the inferior turbinate. In adulthood, depending on the shape of the skull the length of the nasolacrimal duct is from 14.0 to 26.0 mm. Its diameter increases almost twice. The nasal orifice in 5 of 10 cases has a slit-like shape, and in 6 cases it has almost round shape. In the medial wall of the maxillary sinus in the region of its anterosuperior angle is the longitudinal elevation of the nasolacrimal canal, which is directed down, laterally and back. Its length is equal to 14.0 ± 0.4 mm, its width is


48

equal to 8.0 ± 0.2 mm, the thickness of the bone in the elevation is 1.0 ± 0.02 mm. In elderly and senile age topographic and anatomical interrelations of nasolacrimal duct do not change. Its lumen increases slightly due to the mucosal atrophy.

Conclusions. 1. The anlage and formation of the nasolacrimal duct begins in the embryo of 12.0-14.0 mm PCL.

2. The shape and topographic location of nasal duct orifice changes with age.

Prospects for further research. It is planned to perform further studies of blood supply, innervation and lymph flow from these structures.

References: 1. Патент на корисну модель № 83418

(Україна), МПК (2013.01) A61B 5/00, Спосіб визначення морфологічних параметрів носо-сльозової протоки у плодів та новонароджених / Кузняк Н.Б., Бойчук О.М., Сикирицька Т.Б.; заявник і патентовл. Буковинський держ. мед. ун-тет. № заявки u 2013 02943; заявл. 11.03.2013; опубл. 10.09.2013, Бюл. № 17. – 4 с.

2. Логвінова О.В. Фізична реабілітація хворих на короткозорість / О.В. Логвинова, Г.В. Шищук, І.В. Нехаєнко // Актуальні питання теоретичної та клінічної медицини: Міжнар. наук.-практ. конф.

(Суми, 10-12 квітня 2013 р.): матер. конференції. – Суми: СумДУ, 2013. – С. 213-214.

3. Черкунов Б.Ф. Болезни слезных органов / Черкунов Б.Ф. – Самара: Перспектива, 2001. –296с.

4. Chaudhary M. Bacteriology and antimicrobial susceptibility of adult chronic dacryocystitis/ M. Chaudhary, А. Bhattarai, S.K. Adhikari // Nepal. J. Ophthalmol. 2010. -Vol. 4.- Р. 105–113.

5. Тейлор Д. Слезные органы Детская офтальмология / Д. Тейлор, К. Хойт; пер.с англ. - М.: «Издательство БИНОМ», 2007. – С. 99-103.

6. Сайдашева Э.И. Инфекционные заболевания: Избранные лекции по неонатальной офтальмологии / Э.И.Сайдашева, Е.Е.Сомов, Н.В. Фомина – СПб.: Издательство «Нестор-История», 2006. – С. 188-201.

7. Razavi M.E Bacteriological evaluation of adult dacryocystitis in Iran / M.E. Razavi, Ansari–M.R. Astaneh, M. farzadnia // Orbit.- 2010.- Vol. 29.- P. 286–290.

8. Shah C.P comparative bacteriological profile and antibiogram of dacryocystitis / C.P. Shah, D.A. Santani // Nepal. J. Ophthalmol. – 2011. – Vol. 6. – P. 134–139.


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UDC 616-073.756.8

Kravchuk S.Yu., Higher State Educational Institution of Ukraine “Bukovinian State Medical University”, Department of Oncology and

Radiology, Chernivtsi, Ukraine, [email protected]

Kuftiak V.V. Chernivtsi Regional Clinical Oncology Center, Chernivtsi, Ukraine

OPTIMIZATION OF USING OF COMPUTED TOMOGRAPHY METHODS

Abstract. Analysis of methods of computed tomography with intravenous contrast and without contrast according to the radiation exposure, diagnostic value, invasiveness and cost was performed. According to three of the four diagnostic criteria method of computed tomography without i/v contrast prevails. Key words: computed tomography, methods.

Introduction. There are two main methods of computed tomography (CT): CT with intravenous contrast, or CT with contrast image enhancement, and CT without contrast, or CT native. An undeniable advantage of CT with contrast image enhancement has been the dominant view for a long time and it is still so. Meanwhile, with the advent of new diagnostic techniques, including not only radiation, it is important to periodically analyze and rethink pros and cons of these methods of CT, especially taking into account their ionizing effect on the subject.

Objective: To аnalysis of the advantages and disadvantages of the two main methods of computer tomography.

Materials and methods. 237 computer-tomographic inspections, carried out in 2015 with ambulatory and hospitalized patients of Chernivtsi Oncology Center, among which there was 108 inspections by CT with intravenous contrast, have been studied. Besides of computed tomography other radiation (radiography of the chest cavity, ultrasound diagnostics of organs of the abdomen and pelvis), laboratory and instrumental (fibergastroduodenoskopy, bronchoscopy, colonoscopy) studies, followed by biopsy and histological examination of biopsy material, were performed with patients.

Results and discussion. The comparative analysis of the computed tomography both with and without intravenous contrast has been carried out. When choosing a method of

radiation diagnostic the following criteria are commonly used: 1) radiation safety; 2) informativeness; 3) the invasiveness and onerousness for the patient; 4) cost and availability.

Today the concept ALARA (as low as reasonably achievable) is globally applied, which means that for each radiological procedures, including CT, the lowest possible dose must be used [2, 5]. This leads to constant looking for opportunities of reducing radiation exposure to the patient. Thus, resource Eurosafe imaging created by the European Association of Radiologists (ESR) informs patients and radiologists about the risks of radiological procedures and ways to reduce the radiation exposure during the CT.

Earlier, at the computed tomography the amount of radiation received by the patient was evaluated by the tables, contained information on approximate amount of absorbed radiation when examining different parts of the body [1]. Nowadays, the amount of radiation received by the patient during the procedure, is mathematically calculated more accurately due to the presence of function Patient Protocol in modern CT scanners.

We compared the parameters of absorbed radiation dose during the whole study of DLP (mGy×cm) in menu Patient Protocol on 20-slice CT device Siemens Somatom Definition AS, which give an indication of the absorbed dose by each patient individually. The effective dose E (mSv) is equivalent to the absorbed dose and is

© Kravchuk S.Yu., Kuftiak V.V., 2016


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calculated by the formula Е=DLP×ЕDLP where ЕDLP is equal to 0,015 for abdominal and 0,017 for the chest cavity, according to the "European Guidelines on Quality Criteria for Computed Tomography" [3].

During the native study of organs of the chest and abdomen the dose absorbed by most patients is about 300-600 mGy × cm, that corresponds to an effective equivalent dose of 5.10 mSv, depending on the patient's weight and size of the survey area.

At the intravenous contrasting the dose increases significantly to an 800-2000 mHr×cm in average at a summation of all doses at phases of contrasting corresponding to the effective equivalent dose of 14-23 mSv and can be even higher when using postponed phase contrasting. Thus, at CT with i/v contrasting the radiation exposure to the patient increases in 2-3 times (Fig.1).

Fig.1. Protocol of patient O., for who the CT of

organs of the chest and abdomen with intravenous contrast was performed.

Of course, the diagnostic value of CT with

contrast image enhancement is higher than the one of native CT. The tumors and metastases, the structure of the organs and vessels are visualized better. That is why for wider implementation of CT with i/v contrasting at early years of this method the directive requirement of leading radiologists to perform only CT with i/v contrasting appeared. This requirement exists today. But even as at CT with i/v contrasting all the morphological and pathological structures are more visible, most of these structures, including the ones of blood vessels, can be seen based on the sufficiently fundamental anatomical knowledge. Often CT

with intravenous contrast bears no additional information necessary to form the correct diagnosis (Fig. 2).

Fig. 2. CT scan of the patient O. with cancer of

cardia of stomach with metastases to the liver and retroperitoneal lymph nodes.

Often, even when detecting pathological

focus using CT with contrast image enhancement, is difficult to the specialist to distinguish malignant tumor from a chronic disease process with absolute certainty. This forces to apply additional instrumental, laboratory and radiation methods, including methods of interventional radiology, particularly puncture biopsy.

The pathomorphologic conclusion in oncology practice is a prerequisite for further successful patient treatment, including radiotherapy and chemotherapy. Therefore, if further histological study of a pathological focus with defined localization, it is needed just to


51

detect the presence of pathological focus, that can be easily performed by modern CT without contrast enhancement in most cases.

With the advent of modern multislice CT scanners, allowing to analyze slices of 1mm thick, highly qualified and experienced specialist in most cases can identify pathology based on direct and indirect signs even without contrast enhancement, and in doubtful cases, can perform further histological study of the material by patient biopsy of pathological focus. It must be remembered that radiologist needs for a comprehensive approach in his conclusions about the nature of the disease, including data of such non-radiation modern diagnostic methods as endoscopy and determination of tumor markers. These methods often help to detect malignant tumors much earlier, not only by CT with the contrast image enhancement, but by other radiation methods of diagnosis, even positron emission tomography, which reduces the role of CT in defining the nature of the process. It should be added that in many cases in the oncology practice the patients are performed CT for the first time in so late stages that diagnosis and prevalence of the process can be carried out without problems even without i/v contrasting.

Also, it should be noted, that having learnt to analyze high quality images of CT with contrast image enhancement the CT specialists do not improve their skills in analyzing the morphological and indirect signs of the disease, forgetting that previously radiologists were able to make correct conclusions by the analysis of naught information of X-ray images only due to the high level of professionalism, synthesis and analysis of clinical and laboratory data, even in the absence of modern methods of diagnosis. Radiation native CT images of organs of chest and abdomen are barely analyzed in textbooks and scientific journals on radiology.

Regarding the invasiveness and onerousness for the patient, the possibility of allergic reactions and even anaphylactic shock during intravenous administration of iodinated drugs, long list of contraindications, including renal failure, severe diabetes, pregnancy, severe general condition of the patient, thyroid disease,

allergy to iodine compounds, etc., this criterion indicates the disadvantage of the use of CT with i/v contrasting compared to the native CT. Moreover, according to the latest data, during the use of iodinated contrast agents at CT enhances radiation DNA damage [4].

Accessibility for people of native CT is higher than the one of CT with contrast image enhancement, because not all CT scanners are equipped with intravenous injectors. Besides, in order to save the disposable catheters for injectors, CT with intravenous contrast is often performed in treatment and diagnostic institutions only on selected days of the week.

The cost of the CT with contrast image enhancement is higher than the one of native CT. The medical management requires taking into account the cost of not only the contrast, but other consumables (disposable syringes, tubes for pumps and patients), not forgetting that the increasing number of CT scans reduces the resource of X-ray tube, which in this case uses its resource faster, because at CT with i/v contrasting the same area of the body is scanned for several times.

Regarding the current situation in medical institutions of Ukraine, such service as CT of different parts of the body without contrasting is available in all the private diagnostic institutions, and its price is lower than the one of CT with contrast image enhancement in several times, and these data are available Internet for everyone. Native CT is widely used in the practice of public health institutions of Ukraine, including ChRCOC. In the early stages of the CT the requirement to implement it in the study of internal organs only with i/v contrasting was probably justified from the standpoint of the broader implementation of this method in practice. However, the existence and widespread use of native CT in medical practice force us to take into account the needs of practical medicine by wide informing general practice doctors about the advantages and disadvantages of different methods of radiation diagnosis, such as native CT and CT with contrast image enhancement. For example, PET-CT has even greater diagnostic value in detecting tumors than CT with i/v contrasting, but it does


52

not give us the right to object to the latest diagnostic methods in oncology practice. For example, PET-CT has even greater diagnostic value in detecting tumors than CT with i/v contrasting, but it does not mean it needs to object to the latest diagnostic methods in oncology practice.

Thus, a comparative analysis of native CT and CT with i/v contrasting indicates the advantage of native CT in three of the four major diagnostic criteria. The task of radiologist is to more fully and fairly inform other doctors about the pros and cons of each method of radiation diagnosis, including computed tomography, thus expanding their choices. Often radiologists narrow the choices of patients and other doctors who appoint CT to these patients by their categorical necessity in CT with contrast image enhancement in order to analyze images with better quality with more diagnostic information.

Due to the fact that a good doctor should choose the method and amount of diagnostic procedures taking into account the views of the patient, explaining to him all the advantages and disadvantages of the method, there is no assurance that after bringing relevant information to the patient and inviting him to make a choice, he chooses CT with i/v contrasting, especially because of its radiation exposure, which in several times higher than the one of native CT. Moreover, in the absence of a system of health insurance and a decrease in living standards in Ukraine, the patient often chooses a cheaper method of diagnosis.

In addition, in terms of oncology clinic where a significant number of patients receive radiation therapy the radiation dose excess may cause to radiation reactions in some patients such as leukopenia, anemia, etc. In many cases, the advantages of the diagnostic capabilities of CT with contrast image enhancement are offset by the possibility of using the whole range of other non-ionizing research methods, including laboratory and instrumental ones, most of which should be appointed to the patient by the doctor according to the diagnostic protocol of

making the pathomorphologic diagnosis. Also, we should not forget that in doubtful cases it is always possible to conduct CT with contrast image enhancement even after native CT, as the fold difference in radiation exposure of these methods leads to the radiation exposure of CT with contrast enhancement is not significantly different from the one of complex of both CT methods.

Conclusions. 1. CT with intravenous contrast compared to CT without contrast has greater diagnostic value, but is more onerous procedure, has a higher cost, lower availability and has more radiation exposure for the patient. 2. In each case the complex approach to choose the method of computed tomography is necessary, taking into account the current trends in modern methods of diagnosis and the mentioned above selection criteria. 3. CT without i/v contrast is widely used in clinical practice in Ukrainian medical institutions, which leads to the need of developing of radiation semiotics of this radiation diagnosis method for training specialists in computed tomography.

References: 1. Гацкевич Г.В. Проблемы контроля

дозовых нагрузок на пациентов при рентгеновской компьютерной томографии / Г.В. Гацкевич, С.А. Хоружик // Украинский радиологический журнал. – 2010. – № 2. – С. 166-168.

2. Дёмина Э.А. Лучевая диагностика и канцерогенный риск (взгляд радиобиолога) / Э.А. Дёмина // Променева діагностика, променева терапія. – 2015. – № 1. – С. 72-77.

3. European Guidelines on quality criteria for Computed Tomography. Report EUR 16262, Luxemburg (1999). [Електронний ресурс] Режим доступу: http://www.drs.dk/ guidelines/ct/quality/download/EU_html.zip.

4. Intravenous Iodinated Contrast Agents Amplify DNA Radiation Damage at CT./ Piechowiak E.I., Peter J.F., Kleb B. [et al.] // Radiology. – 2015, Jun. - 275(3). – P. 692-697.

5. White paper on radiation protection by the European Society of Radiology // Insights Imaging. - 2011. - Vol. 2. - P. 357–362.

http://www.drs.dk/


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UDC 611.819.2:611.813.8]-053.15

Kryvetskyy V.V., Marchuk F.D.

Higher State Educational Institution of Ukraine “Bukovinian State Medical University”, M.H. Turkevych Department of Human Anatomy, Chernivtsi, Ukraine, [email protected]

Kryvetska I.I. Higher State Educational Institution of Ukraine “Bukovinian State Medical University”, S.M. Savenko Department of

Neurology, Psychiatry And Medical Psychology Chernivtsi, Ukraine

Sapunkov O.D. Higher State Educational Institution of Ukraine “Bukovinian State Medical University”, Department of Pediatric Surgery

and Otolaryngology, Chernivtsi, Ukraine

MORPHOGENESIS OF CHOROID PLEXUSES OF LATERAL VENTRICLES OF THE BRAIN IN PREFETAL PERIOD OF HUMAN ONTOGENESIS

Abstract. Correlation between the size of the lateral ventricles and shape of choroid plexus was established. The increase in size of choroid plexuses in prefetal period is determined by more intensive development of the lateral ventricles. Key words: embryo, prefetus, lateral ventricles, choroid plexuses, human.

Introduction. The analysis of the scientific literature shows that the problem of morphogenesis of the lateral ventricles of the brain throughout the prenatal period of human ontogenesis and their morphological characteristics is not properly studied.

Objective: to determine the dynamic changes of shape and size of the lateral ventricles and choroid plexuses of the brain in prefetal period of human ontogenesis.

Materials and methods. The study was conducted on 9 consecutive series of histological sections of embryos and human prefetuses with 5,0- 80,0 mm parietal-coccygeal length (PCL) with the use of microscopy and morphometry methods.

Results and discussion. It was determined that in the fourth week of development (embryos of 5.0 mm PCL) the brain is represented by three brain vesicles. Thus, in this stage, ventricular system is being formed as cavities of the brain vesicles.

Lateral ventricles, as cavities of the forebrain, appear at the beginning of the 5th week of intrauterine development (6.0 embryos PCL) and start as the formation of lateral protrusion of the anterior brain vesicle - telencephalic vesicles, which give a further development to forebrain hemispheres. At this early stage cavities of telencephalic vesicles have

semispheric shape. Their medial sides communicate with each

other and with the dorsal part of the cavity of anterior cerebral vesicle, which later becomes the III ventricle, cavity of diencephalon. With this moment, gradual transformation of this extremely simple structure into a complex system – the definitive lateral ventricle begins. The development of cavities of the brain and lateral ventricles in particular, is closely associated with the development of the relevant parts of the brain. Thus, the accelerated development of telencephalon and hindbrain, causes a quick differentiation of the parts of the brain (formation of five brain vesicles) and their cavities; and a slow growth of the midbrain area maintains its primary tubular shape (Fig. 1).

Dependence on two processes of development of the hemispheres and differentiation of their internal structures is clearly apparent in establishing of the shape of lateral ventricles. Thus, cranio-dorsal increase of vesicles is observed in the end of embryonic period and beginning of prefetal period of development (6 - 7 weeks), and starting from 10 weeks - posterio-inferior direction of growth leads to formation of elongated, and later arcuate shape of lateral ventricles cavities. The median area of the cavity becomes a central part, and anterior and posterior extremities

© Kryvetskyy V.V., Marchuk F.D., Kryvetska I.I., Sapunkov O.D., 2016


54

Fig. 1. Sagittal section of embryo 6,0 mm TKD,

Stained with hematoxylin and eosin. Specimens. Ob. 4,0 Ok. 5,0. 1 – anterior cerebral vesicle; 2 – middle

cerebral vesicle; 3 – posterior cerebral vesicle.

form the front and lower horns, which are located in the frontal and temporal lobes.

The emergence and increase of occipital lobes in the end of prefetal period (prefetuses with 61.0 - 80.0 mm PCL) gives rise to the appearance of posterior horns - a branch of the lateral ventricle's area, located at the transition of the central part into the inferior horn. So, at the end of period prefoetal period, general configuration of ventricle reaches its definitive form. Formation of structure of the walls that bound ventricular cavity, is the second component that influences on their formation.

Primary semispheric ventricular shape is bound by three walls - superior, lateral and inferior, which are formed by structures of anterior cerebral vesicle, walls are smoothly continues with each other. Medially ventricular cavities communicate with each other. But in embryo with 9.0 mm PCL a protrusion of corpus striatum bud on inferior (ventral) wall appears, which provides a spherical shape to the cavity.

In the beginning of prefetal period, the specified enlargement of ventral wall becomes more definite and is seen as a protrusion of both ventral and part of lateral wall. The upper and medial walls of lateral ventricles, in the 8th week of intrauterine development, are formed by pallium. The anterior part of the region, which is

the caudate nucleus' germ, invaginates into anterior part of the cavity of the ventricle, as a rounded ridge, due to its intensive increase in 7 weeks old prefetuses. This formation causes the narrowing of the mentioned area of the ventricle, and establishment of its crescent configuration. Described shape of lateral ventricle persists during 8th week of intrauterine development.

On 10th week hemispheres begin to grow in anterior and dorso-inferior direction, that leads to formation of germs of the frontal and temporal lobes and cavities of the lateral ventricle in these lobes with complete allocation of the lateral ventricle into central, front (anterior horn) and bottom (inferior horn); structures limiting their walls also change. Thus, medial, superior and lateral walls are formed by pallium, i.e. medulla, that forms the mass of the forebrain. The components, that form inferior (ventral) wall, compose lateral part of corpus striatum, which completely transforms into the caudate nucleus, due to uneven growth of hemispheres and their slight rotation, and its lateral inferior position according the medial part. There are insignificant changes in the walls of the anterior horn. The presence of choroid plexus in the anterior horn is insignificant. The surface texture is uneven, it forms many processes of different widths and lengths, which are represented by many vessels of different caliber, located in the loose connective tissue (Fig. 2).

Inferior horn, which gradually becomes conical in shape, bound superiorly with caudate nucleus' tail, medially – by formed gyrus of the hippocampus, and laterally and inferiorly - by medullary substance of temporal lobes.

The germ of posterior horn, which emerged in the late prefetal period, is surrounded by medullary substance of occipital lobes. During the prefetal period the structures that bound lateral ventricles, gradually become similar to the definitive.

Choroid plexus – a part of the lateral ventricles – occurs as an invagination of medial ventricular wall in the end of embryonic period as a simple fold. During the 7th week this fold is located dorsaly to interventricular opening and is connected to the wall with a wide root. On the


55

Fig. 2. Horizontal section of the head of the prefetus TKD 26,0 mm. Stained with hematoxylin and eosin. Ob. 4,0 Ok. 5,0. 1 – cavity of anterior horns of the lateral ventricles; 2 – cavity of third ventricle; 3 –

cavity of posterior horns of the lateral ventricles; 4 – vascular plexus.

periphery there are few protrusions of folds, which are the origin of further branchy structure of plexus. During 8th week significant changes, determined by great amount of protrusions along the whole length of the ventricle in the structure of plexus are observed. Within 9 weeks in medial portion of the temporal bone germ, a fissure is formed, its bottom thins and sags towards the cavity of the temporal lobe. In the specified fissure a fold of vascular mater penetrates, it is covered with ependymal layer of the ventricular cavity wall and initiates the formation of a second, posterior part of plexus. Anterior part of the plexus of the lateral ventricle increases in size. Multiple processes facing different directions сhange the surface, and protrusions, typical to the previous stage of the anterior part of the plexus development, are observed. In the end of the prefetal period the vascular plexus of the lateral ventricles is quite a complex formation. Its plot, located in the central part, fills almost the entire cavity in both longitudinal and transverse directions. The back of it contacts with the plexus of inferior horn of the lateral ventricle. Connection of these plexuses in transition site of central part into inferior horn is determined at the end of prefetal period.

Conclusions. 1. In prefetal period the intensive development of frontal, parietal,

temporal and occipital lobes of cerebral hemispheres, which is accompanied by formation of lateral ventricles and their choroid plexuses occurs.

2. At the end of the prefetal period no clear distinction between the parts of the lateral ventricles was found, internal relief of their walls is mostly smooth. Choroid plexuses are mainly formed in the central part and in the inferior horns of the ventricles, forming numerous processes of medium height and width.

Prospects for further research It is advisable to study the development of vascular plexus and lateral ventricles of the brain in fetal period of human ontogenesis.

References: 1. Дорошкевич Е Ю. Пренатальная

динамика линейных параметров боковых желудочков головного мозга человека / Е.Ю. Дорошкевич, С.В. Дорошкевич // Совр. асп. гистогенеза и вопр. преподавания гистол. в ВУЗе: матер, науч.- практ. конф., посв. 100-летию со дня рожд. проф. Л.И.Фалина // Морфол. – 2007. – Т. 131, № 3. – С. 66.

2. Коржевский Д.Э. Структурная организация сосудистого сплетения конечного мозга эмбриона человека 8-й недели развития / Д.Э. Коржевский, Е.С. Петрова // Морфол. – 2000. – Т. 117, N° 1. – С. 23-28.

3. Решетілова Н.Б. Особливості будови і форми третього шлуночка головного мозку у плодів четвертого місяця внутрішньоутробного розвитку / Н.Б. Решетілова, Т.І. Туліка, Л. І Ковальчук // Таврич. мед.- биол. вестн. – 2006. – Т. 9, № 3 (ч. 3). – С. 153-155.

4. Самоделкина А. А. Структурно-временная организация хориоэпителио-цитов сосудистого сплетения боковых желудочков головного мозга новорожден-ных крыс / А. А. Самоделкина, Л. Г. Сентюрова, В. А. Шаталин // Астраханский медицинский журнал. – 2012. – № 4. – С. 225–227.

5. Халатурник Г.М. Анатомічні особливості IV шлуночка головного мозку та окремих його структур у плодів та новонароджених / Г.М. Халатурник // Бук. мед. вісн. 2003. – Т. 7, № 3. – С. 135-136.

UDC 616.61.- 091.8:616.441- 089.87


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UDC 616.61.- 091.8:616.441- 089.87

Kuzmenko Y.Y.

O.O. Bohomolets National Medical University, Department of Human Anatomy, Kyiv, Ukraine, [email protected]

STRUCTURAL FEATURES OF A RATS’ KIDNEY NEPHRON AFTER THYROIDECTOMY AND PHARMACOLOGICAL CORRECTION

Abstract. We have studied the structural features of a rats’ kidney nephron after total thyroidectomy without hypothyroidism correction and after the L-thyroxine single-drug therapy. The results showed that 100 days after thyroidectomy in the kidney of the rats, which were administered the replacement therapy with L-thyroxine, there was an incomplete resumption of the ultrastructure and some symptoms of degenerative and destructive processes that indicates that the replacement single-drug therapy does not prevent the development of destructive changes in all components of a kidney nephron, but significantly slows their progress. Key words: nephron, hypothyroidism, L- thyroxin, rats, thyreoidectomy.

Introduction. The relevance of hypothyroidism problems in clinical practice of doctors of various specialties is due to the fact that the deficiency of thyroid hormones necessary for normal functioning of virtually each cell, severe disturbances in all the organs and systems develop, and the prevalence of this disease is about 2%, and in some age groups (elderly women) can reach 6-8% [1, 2]. Therefore, the problem of preventing the development of pathological changes in hypothyroidism acquires an increasing importance. The replacement pharmacotherapy based on a treatment with L-thyroxine, does not provide a full quality of life of these patients [3,4]. Experimental studies in recent years have shown that a treatment of thyroidectomized rats with L-thyroxine does not prevent the formation of morphological and functional changes in the heart, ovaries, brain, and only prolongs the process [5,6]. There are not any data in the literature about the structural changes in the kidney in hypothyroidism and after its correction with L-thyroxine.

Objective: to study the ultrastructure of the kidney nephron after total thyroidectomy in the rats that did not receive replacement therapy and after the single-drug therapy with L-thyroxine.

Materials and methods. The study was conducted on 20 white outbred female rats weighing 180-200 g. The experimental animals were kept and used in accordance with "common ethical principles of animal experiments." The animals were simulated a condition of overt hypothyroidism through the total thyroidectomy [7]. The thyroidectomized

animals were administered L-thyroxine (Pharmak, Ukraine), at a dose of 10 mg / kg per os. Their hypothyroidism was controlled by a determination of free thyroxine rate in the blood plasma of animals using ELISA. The animals were decapitated 100 days after the operation under light ether anesthesia according to the requirements for the removal of animals from an experiment.

Portions of the renal cortex, fixed in 2.5% glutaraldehyde solution buffered with phosphate, with final fixation in 1% OsO4 solution and treated in accordance with the conventional technique served as a material for electron microscopic studies. The morphometric analyze was performed using a semi-automated device for processing graphic images. The resulting digital data were processed by the method of variation statistics using Student's t test. The results were considered reliable at P <0.05.

Results and discussion. A prolonged deficiency of thyroid hormones causes changes in all structural components of the nephron: there are only small and medium-size renal corpuscles, as a result, the average area of a cut of the renal corpuscles in the choroid glomus and the space between the visceral and parietal layers of the capsule is smaller than in the controls (Table 1). Excess of the parameters in volume and quantitative capillary density over the control figures against the background of a significant decrease in renal corpuscle sizes, is only possible with the proportional reduction of the volume and the number of capillaries themselves. The area of the capillaries is also statistically lower (Table 1).

© Kuzmenko Y.Y., 2016


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Table 1 Morphometric parameters that characterize the changes in the glomerular apparatus of the kidneys of rats 100 days after thyroidectomy (I) and single-drug therapy with L- thyroxine (II)

Area of the RC * 102 micron2

Area of the СG, * 102 micron2

Area of the space,

* 102 micron2

Volume density of

capillaries in the RC, %

Quantitive density of

capillaries in the RC* 10-6 micron3

Area of capillaries * 102micron2

Shape factor in

capillaries

control 598,5638,28 524,0341,14 74,526,8 16,90,90 13,691,66 3,150,17 0,730,01 I 367,2042,42 * 325,1939,37 * 42,005,8 * 22,023,06* 39,352,95* 1,840,16 * 0,630,01 II 432,5937,00* 329,2232,46* 103,3711,4* 22,371,22* 23,613,55* 2,450,43* 0,680,01

The worn areas of endothelial cells with

diaphragmed and non-diaphragmed fenestra alternate with the protrusions of the cytoplasm, which frequently take the form of microclasmatose appendixes. The electron-dark material is stored in the the electron-dense cytoplasm. The basal membrane generally maintains a uniform thickness, and in places of capillary bifurcation where the mesangial cells are located in the control group, it is thickened. The cytoplasm of the mesangial cells is electron-dense and the nuclei are rare.

The shape and contents of the podocytes processes change. Most cytotrabecula acquire an increased electron density, the number of their organelles reduces; mitochondria are usually swollen, with visualised matrix and crista. Myosin-positive deposits increase in the cytoplasm, which is typical for cytopodies. The quantitative parameters indicate substantial restructuring in podocytes too: the average area of cytotrabecula and cytopodies reduces in comparison with the control, they become more elongated, as evidenced by the shape factor

(Table 2). The number of cytotrabecula and the volume that they occupy in the volume unit of the glomerulus, are lower than in the control group (Table 2). The number of cytopodies and their volume in the volume unit of the glomerulus are not different from the control values (Table 2).

But considering the fact that the average glomerular area values decrease in this period of observation, we can talk about a significant decrease in the total number of cytotrabecula and cytopodies. The prevalence of degenerated podocytes and changing their quantitative parameters are indicative of the death of some of these cells. It is confirmed by the fact that the number of glomerular capillaries decreases in the same period of observation.

Most of the proximal tubules have fragmented and desquamated microvilli. Almost all the epithelial cells that line the tubules, have lost their inherent form; basal labyrinths do not have a parallel orientation; mitochondria are randomly located in the cytoplasm and their number, as well as that of the biosynthetic

Table 2 Changes in quantitative indices of rats’ podocytes 100 days after thyroidectomy (I) and single-

drug therapy with L-thyroxine (II) Volume density of

cytotrabecula in the RC, %

Quantitative density of cytotrabecula in the RC, * 10-2/microns3

Area of cytotrabecula microns2

Shape factor in cytotrabecula

control 10,211,22 1,140,19 6,291,05 0,480,01 I 7,660,87 0,580,20* 4,731,02* 0,350,01* II 10,392,42 1,060,57 5,701,03 0,550,01* Volume density of

cytopodies in the RC, %

Quantitative density of cytopodies in the RC, * 10-2/ microns3

Area of cytopodies, microns2

Shape factor in cytopodies

control 3,400,23 24,35,6 0,150,01 0,470,01 I 3, 260,16 25,222,90 0,110,006* 0,420,01

II 3,470,53 25,595,04 0,150,008 0,530,01


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organelles reduce. Dark and cleared lysosomes are common in the cells, whereas other structures involved in the transcellular reabsorption of proteins are observed in a small number. There are some tubules, where epithelial cells are in different stages of apoptosis up to the formation of apoptotic bodies. The basement membrane varies in thickness: thinned sections alternate with sections where it is thickened and sometimes destructively changed.

In the distal tubules the epithelial cells also change the ultrastructure, but the severity and extent of these changes is less pronounced than in the proximal ones. The changes are manifested primarily by vacuolization of cytoplasm, by penetration of basal plasmalemma invaginations deep into the cytoplasm and expansion of these invaginations so that the cell seems to be split into pieces. It should be noted that the damaged cells both of the distal tubules and the proximal ones are often located in a cluster way.

100 days after thyroidectomy the rats receiving L-replacement therapy with thyroxine had an average area of the renal corpuscles statistically higher than in the animals without corrective therapy, but it did not reach the control values (Table 1). The choroid glomus area remains statistically the same type with the value in rats treated with L-replacement therapy with thyroxine, whereas the space area exceeds the same figure in the controls (Table 1). Thus, an increase in the area of the renal corpuscles is due to the increase in the size of the space between the parietal and visceral layers of the capsule. Like the area of the renal corpuscle, the average capillary area value did not differ in the animals either (Table 1). An excess in the value of quantitive density of capillaries in the animals treated with replacement therapy with L-thyroxine, was most likely due to differences in renal corpuscles areas in the two groups, but not with changes in the total number of capillaries.

The lumen of most glomerular capillaries is open, the coagulated plasma is practically not seen. There is a cytoplasm protrusion in the worn peripheral areas of endothelial cells. The fenestra are unevenly placed in the cells: continuous endothelium is alternated with areas with a large number of fenestrae. The changes in the glomerular basement membrane are not

visualized. Mesangial cells do not have any significant damage. The cytoplasm is of a moderate electron density, preservation of organelles is more pronounced, and the number of myosin-positive matter is less in comparison with the previous experimental group in the body of the podocytes and their cytotrabecula. The size and the quantitative density of cytotrabecula do not differ from the control values, whereas the form factor indicates their differences both from the control animals, and from those with hypothyroidism (Table 2). They are not only less flattened than in the latter ones but more oval than in the control group. It is due to the fact that some cytotrabecula with clear cytoplasm protrude into the urinary space and seem to contain some edematous fluid. It should also be noted that adjacent podocytes are located rather close to each other and, as a result, the urinary space is narrowed. There are no epithelial cells in the proximal tubules in the final stages of apoptosis. The cells contact closely between each other via the slit and tight contacts. The microvilli which are in contact with endocytic structures of various shape, size and density are well preserved in them. Further, in the basal direction lysosomes are located. Basal labyrinths do not keep the parallelism in all the cells, in some places they are oriented along the basement membrane, so are the mitochondria. The latter are located throughout the cytoplasm, they vary in size, have a moderate electron density matrix, clearly structured outer membrane and cristae. The number of the latter is slightly decreased compared to the control (Figure). In the distal tubules the epithelial cells have signs of active biosynthetic processes: they contain a rounded nucleus with evenly distributed chromatin, a significant number of mitochondria, Golgi tubules, secretory granules. The latter ones, unlike those in the hypothyroid rats are not increased in size (Figure B). Dense contacts are mostly located close to the apical surface and have a great extent in some places. The difference between these cells and those in the control is thei small amount and sometimes complete disappearance of microvilli as well as a disorientation of basal protrusions.

Some areas of thickening, destruction, edema of the basal membrane in the tubules of the nephron, as well as cellular detritus in the interstitial space are indicative of an incomplete


59

А

B

Figure. The kidney of a thyroidectomized rat, treated with L-thyroxine. А – A fragment of the

proximal tubule. B – A frafment of the distal tubule. Electron microscopy micrographs. Magn.: А. –

24000, B. – 6000.

resumption of the ultrastructure and the presence of dystrophic and destructive processes in the kidney after monotherapy.

Conclusions. 100 days after thyroidectomy in rats which received replacement therapy with L-thyroxine, in the kidney there is an incomplete resumption of the ultrastructure and the presence of dystrophic and destructive processes. The results reveal some of the mechanisms of functional disorders in the

kidney with hypothyroidism even against replacement monotherapy and dictate the need for new treatment regimens.

Prospects for further research. In the future, it is advisable to study the ultrastructure of the kidney nephron after total thyroidectomy in rats which received integrated treatment with L-thyroxine and calcitonin.

References: 1. Фадеев В.В. Современные концепции

диагностики и лечения гипотиреоза у взрослых / В.В. Фадеев // Проблемы эндокринологии. – 2004. - Т. 50, № 2. - С. 47.

2. Карлович Н.В. Гипотиреоз: современные представления о коррекции и мониторинге / Н.В. Карлович, Т.В. Мохорт // Медицинские новости. - № 11. - 2004. – С. 32-34.

3. Моргунова Т.Б. Заместительная терапия гипотиреоза препаратами тиреоидных гормонов – один гормон или два?: Обзор / Т.Б. Моргунова, В.В. Фадеев // Проблемы эндокринологии. - 2005. - Т.51, №1. - С. 53-56.

4. Фадеев В.В. Проблемы заместительной терапии гипотиреоза: современность и перспективы / В.В. Фадеев // Клиническая и экспериментальная тиреоидология. – 2012. – Т. 8, № 3. – С. 17-29.

5. Preservation of renal function by thyroid hormone replacement therapy in chronic kidney disease patients with subclinical hypothyroidism / D.H. Shin, M.J. Lee, S.J. Kim [et al.] // J. Clin. Endocrinol. Metab. – 2012. – Vol. 97. – Р. 2732-2740.

6. Петренко В.А. Морфофункціональний стан передсердних кардіоміоцитів щурів при корекції гіпотиреозу L-тироксином та в поєднанні його з кальцитоніном / В.А. Петренко // Український науково-медичний молодіжний журнал. – 2007. – № 3. – С. 16-20.

7. Патент №27821, Україна, МПК G09B23/28(2006.01) Спосіб моделювання гіпотиреозу у щурів // Стеченко Л.О., Петренко В.А., Бик П.Л., Кузян В.Р., Куфтирева Т.П.; Національний медичний університет ім. О.О. Богомольця-№u200708689; Заявл. 30.07.2007.


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UDC 616:89-008.44-092]-053.2

Burbela Е.І., Volianska L.А.,

Kubei І.V., Vorontsova T.O.,

Synytska V.O. SHEE “I. Horbachevsky Ternopil State Medical University of Ministry of Health of Ukraine”, Ternopil, Ukraine,

[email protected]

ASSESSMENT OF ANXIETY IN PERSONALITY STRUCTURE OF CHILDREN SUFFERING FROM BRONCHIAL ASTHMA

Abstract. Design of the study included 121 children with asthma and 226 healthy school children aged 8 to 17, who were assessed for anxiety by Spielberger-Khanin test(1976) and anxiety at school by Philips questionnaire . The aim of the study was to evaluate the personality and psychological components of broncho-obstructive symptoms in patients with asthma. The contingent under study showed disorders in personal anxiety in 97.52% of children and situational one in 78.51%. The indices of the reactive anxiety in the experimental group were slightly lower than of the personal one, but reliably higher than in the control group. The results indicate that children with asthma aged 8-10 years are the most sensitive to stressful school situations and factors. There was a tendency to reduce the importance of the school environment factors with pupils’ growing up. Senior pupils and those from graduation classes are better adapted to the school environment. Obviously, the human factor is important in the severity of school anxiety, as peers and teachers, considering the serious illness of children with asthma, treat them more indulgently and are less demanding towards them as evidenced by medium and low scores on scales of school anxiety. Key words: anxiety, bronchial asthma, children

Introduction. Bronchial asthma (BA) and allergic diseases have become today "a global challenge of modern times" due to considerable prevalence among children and adult population of the planet. From 5 to 10% of children suffer from this disease every year and this figure is growing, urging an ongoing scientific interest in studying etiopathogenetical aspects of asthma [1, 2, 3].

Respiration is the only spontaneous manifestation of a newborn baby independence. Physiological characteristics of breathing are directly connected to its emotional state, and this relationship persists throughout the life. Emotional factors alone can not create sufficient conditions for a disease, but they activate the broncho-obstructive process in biologically prone person [4]. Broncho-obstructive (asthmatic) attack is a spasm of the bronchioles, which can be caused both by allergen exposure and by emotional factors [5]. Asthma is characterized by an action pattern-

bated breath , which is formed by nature, as a defense mechanism in response to an episode that caused fear, to an unexpected event or a premonition of the unknown.

Objective: to study the personal and psychological components of broncho-obstructive symptoms in the schoolchildren with asthma.

Materials and methods. We have examined 121 children with asthma who had been hospitalized in pulmo-allergy department of Ternopil childrens' hospital during 2012-2015. The control group consisted of 226 adolescents, 76,55% (n = 173) of whom were virtually healthy at the time of examination, secondary school students from Ternopil and Terebovlya, and 23,45% (n = 53) – from the villages of Ternopil and Terebovlia districts. The average age of patients was (12.98 + 2.80) years, children in the control group (12.36 ± 2.80) years. Among the patients with asthma in all age groups the number of boys prevailed over that of girls by

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about half, and in the 3rd group the girls were only ¼ part. The criterion for inclusion in the study was possibility and willing to answer questions of the proposed questionnaires. The criterion for exclusion among the children in the control group was identifying background diseases and comorbidity among pediatric patients.

The clinical status of children with asthma was assessed on the basis of anamnesis, physical examination findings, spirometry (peakflowmetry). Asthma severity assessment

was conducted in accordance with the recommendations of the international Global Initiative for the diagnosis and treatment of asthma GINA-2015. The level of BA controllability was evaluated every three months of treatment in accordance with the step therapy. The majority of the observed cohorts, 76,03% (n = 92) was diagnosed with partially controlled disease. Not controlled character was found in 10,74% (n = 13) and the remaining 13,23% (n = 16) in the controlled ones (Figure).

Figure. Level of BA control in the experimental groups, where 1 – controlled, 2 – uncontrolled, 3 – partially controlled.

The majority 77,68% (n = 94) of the observed children were diagnosed with atopic form of the disease, 2,47% (n = 3) with nonatopic and the remaining 19,83% (n = 24) with a mixed form. The dominant course of BA was persistent one in 92,56% (n = 112), including the mild form in 14,05% (n = 17), moderate one in 75,21% (n = 91) and a severe course in 3, 31% (n = 4). Intermittent asthma was diagnosed in 7,44% (n = 9).

All the children under observation were evaluated as to their emotional status by psychological tests and surveys:

- Assessment of anxiety in the structure of personality by Spielberger-Khanin test (1976) [7];

- Assessment of anxiety at school by Philips questionnaire [7].

Results and discussion. Even at the dawn of medicine as a science of healthy and sick people doctors noticed a connection between the features of the individual, their psychological lability and presence of a pathology. Classical psychosomatic diseases today include seven nosologies. Bronchial asthma is one of the "Chicago seven" by F. Aleksander. For all that, there is no single interpretation of the pathogenesis of asthma today. Several modern studies consider etiopathogenetical chain of chronic diseases in general as follows: stress factors – personality – psychological type – somatotype – system-organ deficiency and joining another sixth unit – resistant mechanism of pathological condition (by N.P. Bekhtiereva ), which


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close it in a "pathological circuit" of self sustained inertial system with secondary inactive compensatory pathological structures at each level [8]. The most common implementation of somatic symptom disorder through the respiratory system organs is a system hyperventilation syndrome. The psychogenic factor (anxiety) disrupt breathing, causing hyperventilation. An increased lung, alveolar ventilation leads to persistent biochemical shifts, excessive removal of СО2 from the body, hypocapnia with decreased partial pressure of СО2 in alveolar air and oxygen in arterial blood, as well as respiratory alkalosis. These changes contribute to the formation of pathological symptoms such as muscle-tonic,

sensitive and other disorders that enhance mental disorders and form a pathological circuit. [9] The growth of respiratory and heart rates, an increase of the minute volume of blood circulation, an increased blood pressure, disorders in acid-base balance, an increase in the overall excitability, reduced sensitivity threshold are all physiological symptoms of anxiety [10]. With the duration of the anxiety, the first physiological manifestations in case of recurrency gain constant nature, i.e. the changes in organs and systems gain some signs of the disease. Anxiety becomes somatic. Analysis of the study of personal anxiety in adolescent patients with asthma found no disorders in only 2,48% (n = 3) (Table)

Table Comparative characteristics of anxiety at the time of examination in children with asthma and

healthy pupils.

Personal anxiety

groups 1st , n=31 2nd , n=46 3rd , n= 44

anxiety high n=23

moderate n=7

low n=1

high n=26 moderate n=19

Low n=1 high n=22

Moderate n=20

low n=2

Children with BA,

%

74,19* 22,58 3,22* 56,52* 41,31 2,17* 50,00* 45,45 4,55*

groups 1st , n=72 2nd , n=82 3rd , n=72

high n=4

moderate n=26

low n=42


low n=44

high n=4

moderate n=31

low n=37

% 5,56 36,11 58,33 1,22 45,12 53,66 5,55 43,06 51,39

Situational anxiety

groups 1st , n=31 2nd , n=46 3rd , n= 44

anxiety high n=13

moderate n=10

low n=8


low n=17

high n=6

moderate n=20

low n=18

Children with BA,

%

41,94* 32,26 25,80* 23,91* 39,13 54,84 13,64* 45,45* 40,91*

groups 1st , n=72 2nd , n=82 3rd , n=72

high n=1

moderate n=21

low n=49


low n=59

high а n=0

moderate n=18

low n=54

% 1,39 29,16 68,05 3,66 24,39 71,95 - 25,00 75,00 *The difference is reliable compared to the control, p< ,005

The resulting performance of anxiety in our

survey group of the hospital showed mostly a high level of personal anxiety in 58.68%

(N = 71) of surveyed children. It was moderate in the remaining 38,84% (n = 47). Slightly more than half of respondents (n = 78 - 64.46%) showed situational anxiety: a high degree was observed in 23,97% (n = 29), moderate - in 40,49% (n = 49). These figures

were slightly lower than personal anxiety, but much higher compared with the control group. Our findings about the high level of personal anxiety in children with asthma coincide with those of the research by Trzciaska H, Przybylski G, Kozaowski B, Derdowski (2012). [11], which showed the dependence of the level of asthma control on anxiety and depression. The relationship between the severity of


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manifestations of allergic diseases and the presence of anxiety and / or depression were also shown in the study by Noriega N.H. and several other authors [12, 13, 14, 15].

The school environment is one of the major sources of anxiety for the children involved in our study. Fears in relationships between classmates, teachers, a fear not meet the expectations of others, knowledge tests, self-expression, frustration in a need to succeed, the overall school anxiety are crucial at this age. Analyzing the data from questioning our children with asthma by the method of Phillips, we found symptoms of general anxiety in school in all age groups surveyed. In the first group (age - 8-10 years) there was an increase in the average parameters according to almost all scales. Compared to other age groups the overall school anxiety 17,4 ± 3,1 points, fear of self-express 4,52 ± 0,8 points, fear of tests - 4,48 ± 0,8 points, and fear not to meet expectations of others - 4,68 ± 0,9 points were the most pronounced. These figures were in the range of high values for each of the scales. In group 2 the highest rates were recorded in the following three scales - Frustration in a need to succeed 8,76 ± 1,7 points, low physiological stress resistance – 3,69 ± 0,8 points, and fears in the relationship with teachers - 4 24 ± 1,9 points. However, these figures were in an average range of values for each of the above components of school anxiety and personal characteristics of a pupil. It should be noted that in the 3rd group, that is the oldest age group, the highest rates that corresponded to high values were observed on a scale of social stress - 8,89 (± 1,2) points.

Conclusions. 1. The results of a study in anxiety in adolescent patients with asthma found a disorder of personal anxiety in 97.52% of children and situational one in 78.51% of children.

2. The indices of reactive anxiety in the experimental group were slightly lower than of the personal anxiety, but significantly higher than the in the control group.

3. 58,68% (n = 71) of patients with BA were mainly diagnosed with high level of personal anxiety.

4.The results indicate that children with

asthma aged 8-10 years are the most sensitive to stressful situations and school factors.

5.There is a trend to decrease the importance of school environment factors with age.

Senior pupils are better adapted to the school environment. The human factor is important in the severity of school anxiety, as peers and teachers, considering the serious illness of children with asthma, treat them more indulgently and are less demanding towards them as evidenced by medium and low scores on scales of school anxiety.

References: 1. Brozek M, Zejda JE, Kowalska M, Gebus M,

Kepa K, Igielski M. Opposite trends of allergic disorders and respiratory symptoms in children over a period of large-scale ambient air pollution decline. Pol J Environ Stud 2010; 19(6): 1133–1138.

2. Fedortsiv O, Brozek GM, Luchyshyn N, Kubey I, Lawson JA, Rennie DC, Zejda JE. Prevalence of childhood asthma, rhinitis, and eczema in the Ternopil region of Ukraine – results of BUPAS study. Adv Med Sci 2012; 57(2): 282-289.

3. Asher MI, Stewart AW, Mallol J, Montefort S, Lai CKW, At-Khaled N, Odhiambo J, The ISAAC Phase One Study Group. Which population level environmental factors are associated with asthma, rhinoconjunctivitis and eczema? Review of the ecological analyses of ISAAC Phase One. Respir Res 2010, 11:8. Available at http:// respiratory-research.com/content/11/1/8. Acessed May, 25, 2014.

4. Малкина-Пых И. Г. Психосоматика: Справочник практического психолога. – М.: Изд-во Эксмо, 2005. - 992 с.

5. Горшков О.В. Психоэмоциональные и психосоциальные нарушения у детей, больных бронхиальной астмой //Український вісник психоневрології / О.В.Горшков, А.А.Старикова. – 2002. – Т.10, Вип.1. – С.172-173.

6. Райгородский Д.Я. Практическая психодиагностика. Методики и тесты. Учебное пособые. – Самара: Издательский дом «БАХРАХ-М», 2002. – 672 с.

7. Шевченко Ю. С. Концепция комплексной многоуровневой терапии детей и


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подростков (на модели психогенных психосоматических расстройств) / Ю.С. Шевченко // Вопросы психического здоровья детей и подростков. — 2010. — № 1. — C. 19-29.

8. Филатова Е. Г. Гипервентиляционный синдром: этиопатогенез, диагностика и лечение / Е. Г. Филатова // Фарматека. — 2006. — № 7. — C. 1-4.

9. С.С.Каунова, Е.О.Набокова. Особенности проявления тревоги и тревожности у подростков // Интер-медикал – №3. – 2014. – P. 68-72.

10. Analysis of the relation between level of asthma control and depression and anxiety / H.Trzciaska G. Przybylski, B. Kozaowski [et al.] // S. Med Sci Monit. – 2012. – N. 18. – P. 190-194.

11. Phenome-transcriptome correlation

unravels anxiety and depression related pathways / P. Gormanns, N.S. Mueller, C. Ditzen [et al.] // J Psychiatr Res. – 2011. – N. 45. – P. 973-979.

12. Role of immune-inflammatory and oxidative and nitrosative stress pathways in the etiology of depression: therapeutic implications / G.Anderson, M.Berk, O.Dean [et al.] // CNS Drugs. – 2014. – N. 28. – P. 1-10.

13. Atopic disorders and depression: findings from a large, population-based study / L. Sanna, A.L. Stuart, J.A. Pasco, [et al.] // J Affect Disord. – 2014. – N. 155. – P. 261-265.

14. Rhinitis as a risk factor for depressive mood in pre-adolescents: a new approach to this relationship / P. Audino, S. La Grutta, F. Cibella [et al.] // Pediatr Allergy Immunol. – 2014. – N. 25. –P. 360-365.


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UDC 616.329+611.018.82+616.8

Yashchyshyn Z.M., Zaiats L.М.,

Svystak О.D., Yurkiv І.Y.,

Vodoslavska N.Y., HSEI “Ivano-Frankivsk National Medical University”, Department of Pathophysiology, Ivano-Frankivsk, Ukraine,

[email protected]

ROLE OF THE VAGUS INJURY IN THE DEVELOPMENT OF THE CARDIAC ESOPHAGUS DYSFUNCTION

Abstract. During the experiment with a dissected cervical part of the right vagosympathetic trunk on 23 adult cats, we actually simulated partial cardiospasm, a trigger of which, in our opinion, is the loss of preganglionic chain of parasympathetic reflex arch and an expressed autonomy of innervation as well as independence on the central nervous effects of the cardiac sphincter of the esophagus. The results must be considered when performing a surgery on the organs of the thoracic cavity with the aim of preventing disorders of motor activity of the esophagus (especially its abdominal region). Key words: esophagus, neuron, myenteric plexus, ganglia.

Introduction. The esophagus is known to belong to the organs that are under a significant influence of parasympathetic innervation - the vagus nerve [1, 2, 3].

Mechanical compression of the nerve, a trauma or its involvement in the inflammatory process is observed in many diseases of the chest cavity and mediastinum and cause significant changes in the esophagus [4], which are often accompanied by the cardiac sphincter spasm [3, 5, 6].

Based on the above, there is no doubt of the relevance and validity of studying the processes that occur in intramural esophageal interlacement in case of its innervation dissorders.

Objective: to study the changes in the structural elements of the esophageal myenteric plexus (EPMP) and their effects on the cardiac sphincter function after a high dissection of the vagosympathetic trunk.

Materials and methods. The experiment was performed on 23 adult cats of about the same age and weight, which underwent a partial denervation of the esophagus under an ether anesthesia in sterile conditions by dissecting the right vagosympathetic trunk in the cranial department. Time of the experiment was 1, 3, 7, 15, 30, 45, 60 and 90 days.

To study the nervous system and the esophageal bloodstream we used the following methods: 1) Bilshovsky- Gross impregnation of neural elements with silver nitrate; 2) injection with finely ground paints (Paris blue and black

ink) into the esophageal bloodstream; 3) a combined detection of blood vessels and nerve elements of the esophagus with previous injection of blood vessels; 4) morphometric method and correlation analysis.

Results and discussions. On the 1st-3rd day after the vagus dissection the myenteric plexus neurocytes exhibit different tinctorial properties. Some of them become argentophil ones, others – argentophobic ones. It is difficult to identify nerofibrils in their cytoplasm, their nuclei become smaller or bigger, often being located eccentrically, on the move, or some varicose thickenings occur at the ends of the nerve cell processes.

Along with the above-described phenomena in some neurons of the cranial and middle esophagus there is often an acute swelling, and total chromatolysis and karyolysis, hydropic degeneration and vacuolization of cytoplasm, whose nucleus is difficult to distinguish . These changes lead to the death of a large number of neurons. For instance, in the cranial department of EPMP their number decreases by 1.99, in the middle part by 2.45 and in the cardiac department by 1,97 times. The nerve fibers take irregular contours, sometimes they become considerably thinner or thicker. Sometimes we observe a myelin fragmentation and a disintegration of axial cylinders.

The number of glial cells around each of the neurocites remains virtually unchanged at this term of the experiment, although the surface of

© Yashchyshyn Z.M., Zaiats L.М., Svystak О.D., Yurkiv І.Y., Vodoslavska N.Y., 2016


66

the neuron area, which accounts for one gliocyte, is slightly reduced and is in the cranial esophagus 181,5-243,9 microns², in the middle part - 169,4-240 7 microns² and in the cardiac one- 192,7-247,2 microns².

In the bloodstream of EPMP ganglia the changes only start after 3 days and result in uneven filling of blood vessels with injection masses, watering the bloodstream, increasing the size of capillary loops, a decrease in cross-sectional area of blood vessels, which accounts for a conventional unit of the area of EPMP ganglia. It should be noted that these changes in EPMP angioarchitectonics are mainly found in the cranial and middle esophagus, while its bloodstream cardiac segment remains more or less intact.

7-15 days after the operation in all parts of the EPMP there is a significant number of neurons, changes in which indicate the beginning of a degenerative process. Some neurocites impregnate intensively, their outlines are not clear or deformed, the nuclei are not seen, the processes only get coloured at the beginning of their branching or do not contrast at all. On the other hand some nerve cells are poorly impregnated, their nucleus shrinks, deforms, becomes hypo and hyperchromic, it is often shifted to the periphery of the cell. There are "shadow cells" when instead of a nerve cell we can see its residues with blurred contours formed by the products of granular lumpy disintegration of neurofibrils and lysis of nuclei.

The specific gravity of small neurons decreases in the cranial department by 1.33, in the middle one by 1.12 and in in the cardiac one by 1.07 times. The specific gravity of medium neurocytes increases, and large neurons in the cranial department disappear completely.

At this time of the experiment there is a significant proliferative response of glia. The number of glial cells in an environment of neurons reliably increased in all parts of the esophagus just around neurocytes with a transverse diameter of 16-20 microns and 21-25 microns. The surface area of neurons to one gliocyte decreases in all departments of EPMP more than in previous periods. Changes in the microvasculature of the nerve ganglia of EPMP of the cranial and middle esophagus amplified. They are complemented by plethora of the venous link of MCS. There are some disorders in the cardia: the bloodstream is poorly filled with injection masses, we observe the arteriolar

tortuosity and gaps in capillary loops links, the venous part of the blood flow ways is slightly dilated. The percentage of small vessels with a diameter of 10 microns increases due to a decrease of vessels with larger caliber, but these changes in the cardia are minor. Due to this the cross-sectional area of blood vessels, which accounts for a conventional unit of ganglion area decreases by 2,00-2,15 times (in the cardia - only by1.18 times). The bloodstream capacity, which accounts for one neurocyte, decreases in the cranial and middle parts by1.78 and 1.72 times, respectively, but increases in the cardia by 1.59 times. The area of capillary-neuro-cellular contacts reduces.

This period of the experiment is characterized by a conical transition of the extended esophagus in the cranial and middle parts to the constricted one in the cardiac department, accompanied by dysfunction of the cardiac sphincter by cardiospasm pattern.

The process of degeneration of nerve cells and nerve fibers ends by 30 days. There are much fewer degenerative modified structures in ganglia and their decay products and glia proliferative processes continue. Adverse changes in the bloodstream of EPMP continue to grow, as evidenced by the sharp drop in the area of the capillary-neurocellular contacts in the cranial department by 1,37-1,84, in the middle part by1,43-2,08 and in the cardia by 1, 65-1,79 times. On the 45th day of the experiment degenerative changes in ganglia of EPMP stop, regenerative hypertrophy begins instead, continuing to 90 days. It is accompanied by the appearence of hypertrophied, uniformly impregnated neurons with distinct contours, a distinct nucleus and neurofibrils in the structure of ganglia. Processes of such neurons have a uniform thickness and form numerous collaterals.

At this time of the experiment the proliferative response of glial cells subsides and the number of glial cells around each of the neurocytes does not virtually differ from the norm. The index of the neuron area, which accounts for one gliocyte is not significantly different from the norm either.

On the 60-90th day of the experiment the microcirculatory bed of intramural nerve ganglia of the esophagus become normal too. Arterioles and venules that are around them, take their usual appearence. Their contours turn into smooth lines, there are not any deformations in


67

the vessel walls. The caliber of component parts of the venous link of MCS approaches to normal parameters. Morphometric data in this period also indicate a normalization of the blood supply of the components of EPMP and restoration of the capillary-neurocellular relationships.

Functional status of the cardiac sphincter comes to normal starting with the 45th day of the experiment and its complete recovery occurs on the 60-90th day.

Conclusions: 1. We believe that two points are essential in the pathogenesis of cardiac dysfunction of the esophagus: a) loss of preganglionic link of the parasympathetic reflex arc as a result of a damage to the right vagus nerve; b) expressed innervation autonomy and independence on the central nervous influences of the cardiac esophagus, which is confirmed by morphometry in our studies.

2. The distinctive conical transition from the dilated part of the esophagus to the narrowed one is due to the gradual transition from the area with transneurona ganglia denervation of EPMP (departments dependent on the central nervous influences) to the areas where the Dogel type I neurocytes remain due to the autonomous innervation of the department.

Prospects for further research. The etiology of primary disorders of the esophageal motility and in particular, which is the primary factor in the vicious circle of gastroesophageal reflux remains unclear. According to Edwards they are due to the damage of postganglionic neurocytes [8]. We think that lesions in EPMP preganglionic nerve fibers cause them. The question remains arguable and requires more detailed research.

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функциональное значение адренергических волокон блуждающего нерва у кошки / А.Х. Коде // Физиол. ж. – 1989. – Т.35, №6. – С.61-66.

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5. Мосійчук Л.М. Роль гістамінергічної ланки регуляції в морфогенезі уражень стравоходу та шлунка при виразковій хворобі дванадцятипалої кишки, сполученій з гастроезофагеальною рефлексною хворобою / Л.М. Мосійчук // Проблеми військової охорони здоров’я: Зб. наук. праць Укр.військ.-мед.акад.-К., 2006. – Вип.15. – С.286-294.

6. Морфо-функціональні зміни при стенозах стравоходу у дітей та основні принципи їх корекції / О.Г. Дубровін // Зб. наук. праць співробітників КМАПО ім. П.Л.Шупика. – К., 2001. – Вип.10, кн..1. – С.14-21.

7. Стан секреторної та моторно-евакуаторної функції шлунка у хворих на виразкову хворобу дванадцятипалої кишки і вибір методу лікування / О.М.Кіт, А.О.Боб, І.С.Вардинець [та ін.] // Шпитальна хірургія. – 1998. – №1. – С.42-46.

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10. Кузнецов А.В. Моторная функция пищевода при кардиоспазме по данным рентгенокинематографии / А.В. Кузнецов // Клин.хирургия. – 1981. – №10. – С.32-37.

11. Волобуев Н.Н. Избранные главы клинической эзофагологии / Волобуев Н.Н. – Симферополь. 1996. – 36с.

12. Пути улучшения дигностики и лечения заболеваний пищевода / Стручков В.И., Луцев Э.В., Белов И.Н. [и др.] // Грудная хірургія. – 1982. – №5. – С.69-74.

13. Федорова О.Д. Кардиоспазм. – М.: Медицина, 1973. – 148 с.

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UDC: 616.711-003.8-007.17-085.382.018.52/54

Kholodkova Y.L.,

Tsiurupa А.V. Department of Human Anatomy of Odessa National Medical University, Odessa, Ukraine, [email protected]

COMPARISON OF PLATELET-RICH PLASMA AND NONSTEROIDAL

ANTIINFLAMATORY DRUGS EFFECTIVENESS FORCORRECTION OF DEGENERATIVE DYSTROPHIC CHANGES IN THE VERTEBRA AND INTERVERTEBRAL DISCS

Abstract. The paper gives a comparative analysis of the efficiency of platelet-rich plasma and diclofenac sodium, as frequently used non-steroidal anti-inflammatory drug for correcting degenerative dystrophic lesions of the spine. The introduction of diclofenac sodium in short courses did not produce any effect. It has been proved that the introduction of platelet-rich plasma inhibits the development of degenerative processes and promotes the spinal tissue regeneration. Key words. degenerative dystrophic changes of the spine; non-steroidal anti-inflammatory drugs; platelet-rich plasma; correction; intervertebral disc.

Introduction. Degenerative-dystrophic changes of the spine (DDCS) remain one of the most urgent medical problems today. According to WHO the incidence of dorsopathies in the developed countries is pandemic [1], resulting in temporary or permanent disability. Every year around a million patients in Ukraine seek medical attention in connection with DDCS of varying severity and over 16 thousand of them become disabled [2].

DDCS treatment is always comprehensive and includes measures aimed at reducing the load on the spine, physiotherapy, reflex therapy, manual and medication therapies. [3] As to the medication, non-steroidal anti-inflammatory drugs (NSAIDs) are often used in acute periods for the relief of pain attacks and relieving tissue edema. However, among the classic arsenal there are no effective drugs that improve the supply of nutrients to damaged tissue and stimulate tissue regeneration and revascularization.

In recent years, more and more attention of clinicians have been paid to using the platelet-rich plasma (PRP). This method is based on the use of a number of growth factors contained in platelets and involved in neoangiogenesis processes, proliferation and tissue regeneration [4].

Objective: to study of the effectiveness of using the platelet-rich plasma and its

comparison with NSAIDs (diclofenac sodium) for DDCS correction in experimental animals.

Materials and methods. The experiment included 60 mature Wistar, divided into 3 groups: group I - the animals with DDCS; group II - animals with DDCS and a correction with NSAIDs; group III - animals with DDCS and a correction with PRP. A separate group of animals served as an untreated control.

Simulating DDCS pathology in experimental animals was carried out by forming a permanent asymmetric static compression-distension of the caudal vertebral column [5] according to general ethical principles of animal experiments (I National Congress on Bioethics, of 20.09.2001, Kyiv), the Law of Ukraine «Protection of animals against cruel treating» № 27, Art. 230 of 2006, as amended in accordance with the Law N 1759-VI (1759-17) of 15.12.2009, VVR, 2010, N 9, Article 76., as well as the Code of Ethics of the scientists of Ukraine (Ukrainian National Academy of Sciences, 2009).

The animals were taken out of the experiment by euthanasia (inhalation overdose of ether). The results were evaluated 60 days after the beginning of the pathology simulation.

The platelet-rich plasma was produced by its separation from the whole blood on the unit SmartPrep (manufacturer Harvester Corp, USA), the state registration certificate №10179 / 2011 in accordance with the Decree of the Ministry of

© Kholodkova Y.L., Tsiurupa А.V., 2016



69

Health of Ukraine of 08 February 20011, №69. As NSAIDs diclofenac sodium was used (R.P. number UA / 10237/01/01) at the rate of 1 mg / kg for 5 days. NSAIDs and PRP were injected into the base of the tail.

We used the material of the vertebrae and intervertebral discs (IVD) for pathological studies according to standard procedures [6].

Results and discussions. In histological specimens of group I we observed a significant asymmetry of IVD structures and expressed pathological changes in them. On the side of the compression the fibrous ring is significantly thinner compared to that in the intact animals. A significant shift of the vertebral pulp towards distension is observed. Maximum signs of degenerative and dystrophic lesions of the fibrous ring were marked on the compression side. The collagen plates of the fibrous ring looked stratified and flattened, with areas of disorganization and a variety of polymorphic slots. In these areas we observed multiple fiber necrosis foci which sometimes reached the epiphysis. Pathological changes of the vertebral pulp were presented with a decrease in its volume; on the side of its compression we marked impressions of the collagen fibers in the cavity of the vertebral pulp; in some places there was a pronounced vacuolization of notochordial cells and necrosis foci. The height of the epiphyses on the compression side was slightly higher, there were also chondronecrosis foci.

The animals of group II after a correction with NSAIDs also had degenerative-dystrophic changes in the tissues of the vertebrae and the IVD. The degree of asymmetry of the structures was unchanged in comparison with the animals in group I. There was a little lesser degree of collagen fibers stratification and a little larger thickness of the fibrous ring on the compression side, respectively. The number of necrosis foci remained unchanged (Fig 1).

After a correction with the PRP in animals of group III there was a significant decrease in the number of fibronecrosis foci, large thickness of the fibrous ring on the side of the compression, smaller degree of collagen fibers stratification. There was a slight reduction in tissue asymmetry between compression and distension sides. A

slight decrease in the number of foci of notochordial cells necrosis and a change in the height of the epiphysis of the vertebral bodies were also observed. The asymmetry of the IVD vertebral pulp location remained (Figure 2).

Fig. 1. An area of IVD of a rat from group II stained

with hematoxylin and eosin. Magn. х 40.

Fig. 2. An area of IVD of a rat from group III stained

with hematoxylin and eosin. Magn. х 40.

The findings suggest that NSAIDs, without

possessing a regenerative potential are not able to adequately counteract the pathological factors and do not produce significant anti-inflammatory effect in the DDCS. Using the PRP, which contains a great number of growth factors [7], stimulates the processes of regeneration and neoangiogenesis, reduces the severity of DDCS. A comparative evaluation of the effectiveness of NSAIDs and PRP correction


70

showed that the use of PRP reduces the severity of DDCS effectively, stimulating regenerative processes, reduces the degree of ischemic damage and promotes the recovery of the tissue structure.

Conclusions. Using the PRP for DDCS correction has a strong positive effect, while the use of NSAIDs has no significant impact on the course of the pathological process.

Prospects for further research. We are going to carry out more research on the PRP positive effect duration, to elaborate introducing schemes of PRP in the DDCS zone, to study more in details of the formation of vascular network in the DDCS zone. We need to find correction techniques for the location of the IVD vertebral pulp.

References: 1. Bogduk N / Medical management of acute

and chronic low back pain. / Bogduk N, Mc Guirk B // Amsterdam: Elsevler, 2002.

2. Аль-Замиль М.Х. Анализ корреляции морфологических и анамнестических данных при остеохондрозе пояснично-крестцового отдела позвоночника / М.Х. Аль-Замиль, А. В. Мосейчук // Клиническая

неврология. – 2008. – № 4. – С. 7-9. 3. Восстановительное лечение

остеохондроза позвоночника / И.Л. Пшеетаковский, И.П. Шмакова. – Одесса: Астропринт, 2009. – 112 с. – Библиогр.: с. 111 – рус.

4. Robert E. Mars / Platelet-Rich Plasma (PRP): What is PRP and What is not PRP? / Robert E. Mars // Implant dentistry. – 2001. Vol. 10. – P. 225-228.

5. Пат. № 106085 / Спосіб моделювання дегенеративно-дистрофічного ураження хвостового відділу хребта у щурів / Заявл. 18.11.15/ Опуб 11. 04. 16/ Бюл. № 7 / О.Л. Холодкова., О.В. Цюрупа, І.Ю.Бадьїн.

6. Методики морфологічних досліджень : монографія / М54 Багрій М.М., Діброва В.А., Попадинець О.Г., Грищук М.І. ; за ред.. М. М. Багрія, В.А. Діборови. – Вінниця : Нова книга, 2016. – 328 с. ISBN 978-978-966-382-594-6

7. Platelet content and growth factor release in platelet-rich plasma: a comparison of four different systems / G. C. Leitner, R. Gruber, J. Neumüller [et al.] // Vox Sang. – 2006. – Vol. 91. – P. 135 - 139.


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CONTENT:

Khmara T.V., Komar T.V. Individual anatomical variability in the innervation of thigh and calf muscles in human fetuses 3

Sharapova E.N. Morphological and functional changes of rats testes after the action of electromagnetic fields of high voltage with low frequency 7

Romaniuk L.B., Kravets N.Y., Pokryshko O.V., Borak V.P., Oliinyk N.М. Gut microbiota in infants with perinatal pathology of the central nervous system 9

Balaniuk І.V. Reactive response of neutrophilic granulocytes of the peripheral blood in patients with chronic hepatitis C 12

Bodnarchuk Yu.V. Morphological and functional changes and cluster characteristics of hepatocytes in immature rats with streptozotocin-induced diabetes 16

Goroshko O.M., Zamorskii I.I., Drachuk V.M. Influence of antioxidants on the morphological changes in kidneys at experimental acute injury 20

Mazur O.O. role of active forms of oxygen in the aging process (literature review) 24

Vozna H.I. The state of the vegetative provision of cardiac performance in patients with hiv-infection 29

Nekrut D.О. Hyperhomocysteinemia as a factor contributing to the development of non-alcoholic fatty liver disease 34

Pelin R.H., Havrylenko O.O. Spiral computed tomography in the diagnosis of nephrolithiasis 39

Klitynska O.V. Analysis of the dependence of state of local immunity and microbial coenosis in oral cavity on the activity of caries in children residing permanently in conditions of biogeochemical fluorine and iodine deficiency

41

Bambuliak A.V., Boichuk O.M., Honcharenko V.A. Topographic and anatomical features of nasolacrimal duct 46

Kravchuk S.Yu., Kuftiak V.V. Optimization of using of computed tomography methods 49

Kryvetskyy V.V., Marchuk F.D., Kryvetska I.I., Sapunkov O.D. Morphogenesis of choroid plexuses of lateral ventricles of the brain in prefetal period of human ontogenesis 53

Kuzmenko Y.Y. Structural features of a rats’ kidney nephron after thyroidectomy and pharmacological correction 56

Burbela Е.І., Volianska L.А., Kubei І.V., Vorontsova T.O., Synytska V.O. Assessment of anxiety in personality structure of children suffering from bronchial asthma 60

Yashchyshyn Z.M., Zaiats L.М., Svystak О.D., Yurkiv І.Y., Vodoslavska N.Y. Role of the vagus injury in the development of the cardiac esophagus dysfunction 65

Kholodkova Y.L., Tsiurupa А.V. Comparison of platelet-rich plasma and nonsteroidal antiinflamatory drugs effectiveness forcorrection of degenerative dystrophic changes in the vertebra and intervertebral discs

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