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UJR 2004, vol XII, # 4

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2004, vol 12, № 4, page 363

Y. А. Іvаnіv

Capabilities of investigation of right ventricle diastolic function in aortic stenosis using pulsed tissue Doppler ultrasound

Annotation

Objective: To study the changes in longitudinal kinetics of the right ventricle (RV) in response to diastolic dysfunction of the left ventricle and secondary pulmonary hypertension in severe aortic stenosis (AS) using tissue pulsed Doppler ultrasound study (TPD).

Material and Methods: The study involved 14 AS patients (mean age 62.0 ±6.7 years). Ultrasound study of the heart was performed using TPD indices, which characterize both systolic and diastolic kinetics of the RV.

Results: Severe AS with the area of the opening measuring 0.7 ± 0.04 cm2, systolic pressure gradient 67.0 ± 2.8 mm Hg and left ventricle mass index 174.0 ± 7.2 g/m2. The analysis of longitudinal kinetics of the RV revealed considerable changes in its diastolic function, i.e. the velocity of atrial movement was higher when compared with the controls (0.26 ± 0.012 m/s), low Em/Am ratio (0.69 ± 0.052), increased time of early diastolic deceleration (129.0 ± 8.5 m/s), increased is volumetric relaxation (72.0 ± 7.4 m/s). Correlation analysis showed that of all indices of AS severity, only LV mass index correlates reversely with E /A (r = -0.68;p<0.01).

Conclusion: TPD is highly sensitive in evaluation of RV relaxation disorders due to pathology of the left portion of the heart. In patients with severe AS deep changes of diastolic function of the RV myocardium occur, their severity depends on the mass of the left ventricle.

Key words: tissue pulsed Doppler ultrasound study, aortic stenosis, diastolic function, right ventricle.

2004, vol 12, № 4, page 367

Y. О. Vynnyk, М. Y. Nеffа, О. V. Kаzmyruk, І. V. Pyrogovа

The use of diagnostic capabilities of ultrasound and MRI in multimodality treatment for ovarian cancer

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Objective: To study the capability of MRI in determining the degree of dissemination of the tumor process before surgery, and to develop on the basis of the given method a diagnostic algorithm for definition of indications to combined operative treatment in patients with ovarian cancer.

Material and Methods: 146 patients with ovarian cancer were divided into 2 groups: group 1 - 49 patients, in which diagnostic algorithm «clinical examination + ultrasonography + MRI» was used and group 2-97 patients, which diagnostic algorithm «clinical examination + ultrasonography» was used. At estimation of the informativity of diagnostic methods the following parameters were used: sensitivity, specificity, and diagnostic efficiency.

Results: The analysis of findings of sonography and MRI showed, that the maximal inf ormativity at anatomical and differential diagnosis of the degree of dissemination of the tumor process in the patients with ovarian cancer was reached at application of algorithm «clinical examination + ultrasonography + MRI», its diagnostic efficiency was 11 % higher than application of algorithm «clinical examination + ultrasonography».

Conclusion: The application of MRI in patients with ovarian cancer allows to establish the degree of dissemination of the tumor process, which, in turn, promotes planning of the most adequate size of the surgical intervention, in particular, of combined operations.

Key words: cancer of ovaries, diagnostic algorithm, ultrasonography, MRI, combined operations.

2004, vol 12, № 4, page 371

І. P. Vаkulеnko, Т. P. Jabluchаnskа

Comparative characteristic of linear velocity of early diastolic left ventricle filling at permanent atrial fibrillation and sinus rhythm

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Objective: To compare statistical values of linear velocity of left ventricle ( LV ) early diastolic filling at constant atrial fibrillation (CAF) and sinus rhythm (SR).

Material and Methods: The study involved two comparable groups of patients with CAF and SR. CAF group included 17 patients, of them 5 women and 12 men, aged 61 ± 14. CAF duration ranged from several months to 25 years. SR group consisted of 15 patients, of them 6 women and 9 men, aged 62 ± 11. Doppler echocardiography was performed using SIM 5000 plus scanner ( Italy ); linear velocity of LV early diastolic filling was measured. Mean (M), its standard deviation (sd), maximum (max) and minimum (min) values, moda and excess (ex) were calculated. Significance was evaluated using Student's test.

Results: Constant atrial fibrillation differed from SR in statistically higher velocity of early diastolic filling and greater variability of the values. The existing differences reflected faster and more considerable filling of the LV in the phase of passive filling in CAF group when compared with SR.

Conclusion: In CAF, statistical values of linear velocity of early diastolic filling of the left ventricle are of the same order as in SR. In this disease, Doppler echocardiography can be performed in compliance with SR protocol.

Key words: Doppler echocardiography, atrial fibrillation, sinus rhythm.

2004, vol 12, № 4, page 374

L. О. Shkondіn

Complex radiodiagnosis of common cysts of the renal sinus

Annotation

Objective: To define more precisely the capabilities of ultrasound tomography (UST), intravenous urography (IVU), computed tomography (CT), helical CT (HCT) and magnetic resonance imaging (MRI) in detection and differential diagnosis of common cysts of the renal sinus.

Material and Methods: The findings of examination of 103 patients aged 40-77 (31 men and 72 women) with solitary and multiple uni- and bilateral common cysts of the renal sinus (in one or both kidneys) were analyzed. Complex clinical laboratory examination, traditional UST, IVU, MRI, CT and HCT were performed.

Results: It was established that complex radiodiagnosis of common cysts of the renal sinus depended on the number of the cysts, their size, localization, the volume of radiodiagnosis and accuracy of the analysis. Optimal screening technique was UST, IVU being the second stage. If the patient was examined using MRI for the first time, pulsed sequence MSSE of T1 weighted image, RARE T2 weighted image and MR urography should be used. Primary diagnosis of the processes with CT and HCT required contrast enhancement (CE).

Conclusion: Common cysts of the renal sinus are revealed mainly at the age over 50 (86.4%), more frequently in women (70.9%). They can be revealed by chance, without clinical manifestations (65%) or be accompanied by the signs of chronic pyelonephritis, arterial hypertension, pain. An effective diagnostic technique in common cysts of the renal sinus is UST (sensitivity and specificity are close to 100%). Typical for US picture is visualization of asymmetrical in size and location cysts in the sinus. They can have partitions, cannot be joined into a united cavitary system of the kidney and do not respond to diuretics administration. Small cysts are located in the hiatus and mimic hydronephrosis being the cause of diagnostic pitfalls. In case of multiple cysts revealed by UST, which exceed 2- 3 cm on IVU, typical picture of multiple voluminous formations is seen in the sinus of a working kidney, which allows to exclude hydronephrosis (efficacy parameters are close to 100%). But in solitary cysts < 2- 3 cm they are not always revealed with IVU (sensitivity 25%). MRI diagnosis of solitary common cysts in the sinus is an effective method (parameters are close to 100%), because the picture of the lesion is typical and does not depend on the size and localization. In case of multiple cysts, errors are possible (sensitivity 89.9%) because exclusion of RARE T2 weighted pulsed sequence from the investigation and inattentive analysis of MR urography findings can cause erroneous diagnosis of hydronephrosis. CT or HCT without CE cannot be the techniques of final diagnosis of common cysts of the renal sinus because other lesions have similar signs. Only the use of CT and HCT with CE allows to make a final diagnosis of cysts in the renal sinus, exclude their relation to the collecting system (parameter are close to 100%).

Key words: common cysts of renal sinus, diagnostic imaging.

2004, vol 12, № 4, page 382

І. М. Ponomаrov

The efficacy of multimodality therapy for breast cancer in patients of reproductive age depending on the volume of surgical intervention

Annotation

Objective: To study the efficacy of conservative treatment for breast cancer ( ВС ) in patients of reproductive age.

Material and Methods: The study involved 120 ВС patients, of them 64 underwent breast-conserving surgery and 56 Patey's mastectomy. After the surgery, supraclavicular, subclavicular, and parasternal nodes were irradiated at a mean focal dose of 40 Gy. In the patients with breast-conserving surgery, the whole breast was irradiated at a mean focal dose of 55-60 Gy. When axillary nodes were involved, the patients were administered 6 courses of adjuvant CMF polychemotherapy and endocrine therapy.

Results: The lowest values of the studied parameters were observed in patients with the tumor in the inf erointernal quadrant. At breast-conserving surgery, mean duration of relapse-free and metastasis-free periods decreased with the increase in the size of the tumor. The values of 5-year relapse-free survival were somewhat lower at breast-conserving surgery in patients of reproductive age except for the cases when the tumor size did not exceed 2 cm , but this difference was significant only for T2N0M0 tumors. Five-year relapse-free and metastasis-free survival was significantly higher at the initial stages of ВС (T1N0M0) in patients of group 1 and did not differ from that in T2N0M0 and T2N1M0 disease in patients of group 2. These tended to decrease in T1N1M0 tumor.

Conclusion: In patients of reproductive age with stage I-II breast cancer, the indices of survival do not differ between the groups of patients with breast-conserving surgery and mastectomy. The percentage of relapses of stage I-II ВС is higher at breast-conserving surgery in all cases except for T1N0M0 disease.

Key words: breast cancer, breast-conserving surgery, reproductive age.

2004, vol 12, № 4, page 388

L. V. Bаtyuk

The analysis of acid hemolysis parameters and erythrocyte thermal resistance at radiation therapy of malignant tumors

Annotation

Objective: To evaluate the changes in hemolytic and thermal resistance of erythrocytes in cancer patients undergoing radiation therapy (RT).

Material and Methods: Erythrocytes from donor blood and cancer patients (breast cancer, lung cancer, uterine body cancer) obtained before RT and in early terms after it were used. The parameters characterizing hemolytic resistance of erythrocytes, velocity constant and energy of thermal hemolysis activation were determined.

Results: Changes in acid resistance of erythrocytes of cancer patients were observed before RT when compared with donor blood, which was determined by percentage of cell destruction, terms of beginning and ending of hemolysis. Analysis of thermal hemolysis results in cancer patients showed that temperature-induced transition was observed in the erythrocyte membrane at 42-46 ° С.

Conclusion: RT reduces erythrocyte resistance to acid hemolysis, which can be associated with appearance of new population of erythrocytes with renewed membranes. Structural reconstruction of lipid biological layer is observed at temperature below 45 ° С in cancer patients before and after RT. RT causes the changes of protein-lipid interactions in the membranes, which provides approaching of activation energy parameters of cancer patients to those of donors.

Key words: erythrocyte, membrane, acid resistance, thermal hemolysis, activation energy.

2004, vol 12, № 4, page 394

V. Мylаshеnе, G. Prаsmіckеnе, А. Burnеckіs, V. Norkеnе

Correction of the immune and antioxidant systems in patients with stage III rectal cancer during radiation therapy

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Objective: To evaluate the impact of honey and pollen on the immune and antioxidant systems in patients with stage III rectal cancer during radiation therapy.

Material and Methods: This study involved 35 patients with histologically confirmed stage III colorectal cancer. Twenty patients used honey and pollen during radiation therapy (RT), 15 patients did not use bee products. In order to investigate RT-induced attenuation of cell immune response, total leucocyte, lymphocyte, count CD3, CD4, CD8, CD16, CD20 subsets and lymp-hoblast transformation induced by phytohemagglutinin (PHA) were measured in vitro. Percentage count of CD3, CD4, CD8, CD16, CD20 lymphocyte subpopulations were investigated using immuno-fluorescence method.

The level of lipid peroxidation product, malonic dialdehyde (MDA), and activity of antioxidant protection enzyme, catalase (CAT), were studied in patients with rectal cancer. MDA cocent-ration was determined with tiobarbituric acid test using spect-rophometry. CAT activity was evaluated according to formation of colour complex of hydrogen peroxide with ammonium molyb-date.

Venous blood of cancer patients was tested 2 days before radiation therapy (analysis 1), 7 days after beginning of radiotherapy (analysis 2) and 14 days after beginning of radiotherapy (analysis 3) and after the end of radiotherapy (analysis 4).

Results: The immune response was impaired in patients with malignancy, and radiation therapy could exacerbate cancer induced attenuation of immune response. The findings of our investigations showed that during radiation therapy the amount of leucocytes significantly decreased (by 22 %). Percentage and absolute number of total lymphocytes decreased by 24 % and 40 %, respectively. The amount of lymphocytes subsets CD3+ decreased by 14 % , 44 % and CD4+ - by 12 % , 60 % , respectively, in the peripheral blood after 7 days of radiation therapy. Not only the lymphocyte count but also the in vitro lymphocyte stimulation response to mitogen PHA decreased by 25 %.

Lipid peroxidation (MDA) was observed to be higher in the blood serum of patients during RT. The changes of catalase activity were insignificant. However, RT did not change significantly the number of lymphocytes and its subsets in the peripheral blood of the patients who used honey and pollen. Lipid peroxide level (MDA) decreased in the blood serum of colorectal cancer patients applying honey and pollen during RT in comparison with MDA level obtained in patients treated only by RT.

Conclusion: Using honey and pollen positive by influences the immune and antioxidant systems of the patients with stage III rectal cancer during radiation therapy.

Key words: radiation therapy, immune and antioxidant systems, rectal cancer, honey and pollen.

 

2004, vol 12, № 4, page 399

L. І. Symonovа, L. V. Bіlogurovа, C.M. Pushkаr, E. B. Rаdzіshеvskаy

The state of coagulation system in patients with breast cancer after pre-operative radiotherapy

Annotation

Objective: To study the character of changes in the system of coagulation hemostasis in 55 patients with breast cancer before and after pre-operative radiotherapy (RT).

Material and Methods: The study was done in two stages, i.e. before and after the course of pre-operative RT. The state of hemostasis system was studied using electrocoagulography with the account of the most important structural chronometric parameters of electrocoagulogram. Total f ibrinolytic blood activity and paracoagulation processes were determined with biochemical methods.

Results: The analysis of the obtained findings showed that stage 1 of the study was characterized by marked disturbances of hemostasis system. In 50 % of the patients there was hypercoagulation (2 times reduced) fibrin clot formation with high occurence (71.4 % ), which suggested the processes of paracoagulation against a background of increased fibrinolytic activity at sufficiently moderate (33—40 %) incidence of solible fibrin-monomer complexes prior to the surgery. Pre-operative RT provided normalizing coagulation hemostasis both in patients with increased and decreased hemostatic potential. But the signs of disseminated intravascular coagulation were present during the study.

Conclusion: Primary study reveals functional disturbances of coagulation hemostasis in all ВС patients (in 50 % hypercoagulation changes, in the rest hypocoagulation ones); pre-operative RT provides normalizing hemostasis both in patients with increased and dectreased hemostatic potential; the presence of disseminated intravascular coagulation at various stages of the study suggests the danger of thorombo-hemorrhagic complications and requires adminitration of correcting therapy to all cancer patients.

Key words: breast cancer, coagulation hemostasis, fibrinolytic activity, hypercoagulation, disseminated intravascular coagulation syndrome.

2004, vol 12, № 4, page 404

V. А. Vіnnіkov, B. C. Маznyk, A. V. Shеgolkov, О. Е. Іrhа

Protocol for cytogenetic analysis results registration in human lymphocyte long-term cultures after radiation exposure

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Objective: To elaborate an optimized protocol for registration and initial statistical treatment of cytogenetic analysis results obtained at different culture time in human blood lymphocytes after radiation exposure.

Material and Methods: Conventional cytogenetic analysis was carried out in FPG-stained preparations made from 50- and 76-hr cultures of peripheral blood lymphocytes sampled from 15 patients with uterine cancer (stages I—II) after external beam or combined radiation therapy; the programme was created using Rapid Application Development Environment Inprise Borland C++ Builder 6 and database core Borland Database Engine.

Results: The original detailed protocol and its computerised version are proposed for the full-scale analysis of the cytogenetic damage spectrum, which could be observed in long-term cultures of human lymphocytes after radiation exposure. The protocol is based on the evaluation of 30 parameters (various types of chromosomal rearrangements and genomic damage) and allows to calculate yields and to construct per-cell distributions of particular cytogenetic abnormalities or their arbitrary combinations. A special attention was paid to the chromosome fragments (accompanying the exchanges and replicated status). The possibility for choosing the way of data analysis was foreseen, that were either differentially within the particular mitisis or within the combined heterogeneous cell population at the particular culture time.

The practical possibilities of the protocol were demonstrated using the empirical example, which represented the analysis of cytogenetic indices in vitro dynamics in lymphocytes of uterine cancer patients after radiation treatment.

Conclusion: The proposed protocol and its computerized version allow to enhance significantly the efficacy of cytogenetic assay for evaluating the changes of aberrant cell spectrum and studying the chromosome damage transmission through several post-radiation mitoses in the cell population inhomogeneously exposed to ionizing radiation. It also promotes the elaboration of the appropriate criteria for investigating the late genetic instability on the chromosomal and sub-cellular levels.

Key words: chromosome aberrations, genomic damage, lymphocytes, long-term cultivation, radiation exposure.

2004, vol 12, № 4, page 415

N. О. Маznyk, V. А. Vіnnіkov

Calibration dose-response relationships for cytogenetic biodosimetry of recent and past exposure to low dose gamma-radiation

Annotation

Objective: To generate the basic calibration curves for chromosomal biodosimetry of recent and past exposure to low dose y-irradiation using conventional cytogenetic analysis and fluorescence in situ hybridization (FISH) technique.

Material and Methods: Human blood samples were acutely irradiated in vitro to y-rays within dose range 0-1.0 Gy. Cytogenetic analysis was carried in 48 h lymphocyte cultures using conventional method and FISH highlighting chromosome combinations 1, 2 and 4 and 6, 9, 15 and 21 and centromeres of all chromosomes. Stable aberrations were recorded with modified conventional-PAINT descriptive nomenclature, accounting each complete chromosome exchange as an entity. The dose response curve fitting was performed by iteratively reweighted least squares method.

Results: The dose response for both conventionally and FISH-measured chromosome aberration yields showed a good fit to the classic linear-quadratic model. Calibration curve for the full genome dicentrics plus centric rings yield estimated on FISH coincided with that of estimated by conventional analysis, both displaying a linear term about Зх10-2хGy-1 and quadratic term about 8х10-2хGy-2. Excluding incomplete translocations in cells with actual or assumed partial lack of chromosomal material from data treatment provided a dose response for truly radiation-induced translocations, which was very close to that of for dicentrics. The calibration curve for translocations plus insertions in 'stable' cells had a linear coefficient about 1.40х10-2хGy-1 that was 3 times higher than a respective parameter for complete translocations alone. The calibration data showed that the minimum dose, which can be statistically detected by conventional dicentric method with 1000 cells scored, was about 0.14 Gy of acute and 0.20 Gy of chronic y-exposure. The relative sensitivity of FISH biodosimetry with 1000 genome equivalents analyzed varied from 0.22 to 0.31 Gy of acute and from 0.50 to 0.85 Gy of chronic irradiation according to the increase of spontaneous yield of stable chromosome exchanges with persons' age within 20-60-year interval.

Conclusion: For the calibration purposes, both chromosome cocktails used in our survey appeared to be suitable for detecting the true level of aberrations in full genome. The coincidence of the dose response curves for dicentrics and truly radiation-induced translocations confirmed the hypothesis about identical mechanisms of balanced and non-balanced interchromosomal exchange formation in irradiated human lymphocytes. The calibration curve constructed for truly radiation-induced chromosome exchanges in «stable» cells seems to be more appropriate in practical retrospective biodosimetry, in compare with complete translocations alone, especially for chronic exposure conditions.

Key words: dicentrics and centric rings, translocations, fluorescence in situ hybridization, lymphocytes, calibration curves, biological dosimetry.

2004, vol 12, № 4, page 426

B. V. Lеontev, V. V. Оvchаrеnko,

О. І. Romаnov, V. V. Sulіmа

Frequency-contrast characteristics of x-ray visualization systems

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Objective: Theoretical substantiation and experimental study of methods of analytical calculation and experimental determining statistical frequency-contrast characteristics of the system for x-ray visualization as a main quality criterion for revealing low-contrast formations.

Material and Methods: An approach based on the methods of mathematical statistics was used for theoretical calculation of frequency-contrast characteristics of the system for x-ray visualization.

Results: The ratios which allowed to calculate statistical frequency-contrast characteristics of the system for x-ray visualization as a function of values of the parameters of an x-ray unit and its compounds were obtained. The graphs demonstrating the findings of calculations and illustrating the dependence of the quality of observation of low-contrast formations on the exposure dose, the size of the uneven formation, and main parameters of visualization system are presented.

Using a statistical approach, a method of experimental determining frequency-contrast characteristics of the system based on simple measurements followed by their computer processing was worked out.

The correctness of the suggested methods was proved by the results of experimental measurement performed with the use of MADIS mammography unit.

Conclusion: The study allowed to work out and test experimentally the methods of analytical and experimental determining statistical frequency-contrast characteristics of the system for x-ray visualization. Due to the common approach the suggested methods can be used at various types of x-ray visualization. The developed theoretical apparatus can be used for optimizing the systems of x-ray visualization at the stage of a project, the technique for measurement statistical frequency-contrast characteristics allows to include qualitative parameters for comparison and analysis of such systems.

Key words: frequency-contrast characteristics, x-ray visualization, quantum noise, observability, low contrast, spatial frequency, statistical data.

2004, vol 12, № 4, page 431

V. G. Knіgаvko, О. P. Меshеryakovа, E. B. Rаdzіshеvskаy

Mathematical simulation of influence of irradiated cell reparative system saturation on cell survival. Communication 1. Simulation of survival curves in proKaryotes

Annotation

Objective: To build mathematical models of processes of survival curve forming for prokaryotes at exposure to x- or gamma-rays.

Material and Methods: Mathematical simulation, algorithm, programming.

Results: Three models which corresponded to these model suggestions were built and analyzed. Absolute lethal unrepairable radiation lesions (NRL) did not form, but dangerous repairable radiation lesions (DRRL) of one type appeared (model 1); NRL did not form, DRRL of two types appeared (model 2); DRRL of one type and NRL developed (model 3). The models were based on the ideas about saturation of the systems of cell reparation at the highest radiation doses.

Conclusion: Mathematical models of the processes of forming survival curves for prokaryotes which are based on the idea about the possibility of saturation of radiation lesion reparation systems of DNA of the irradiated cells at the dose increase were worked out. The models suggest accessibility of any dangerous radiation lesion for any reparative enzyme complex. These suggestion is true for prokaryotic cells. For the simplest of the discussed models the authors discuss the question about the methods of evaluation of the model parameters.

Key words: mathematical simulation, radiation lesion reparation, survival curves.

2004, vol 12, № 4, page 438

V. G. Knіgаvko, О. P. Меshеryakovа, E. B. Rаdzіshеvskаy

Mathematical simulation of influence of cell reparative system saturation on the survival of these cells.

Communication 2. Simulation of survival curves in euKaryotes

Annotation

Objective: To build mathematical models of the process of forming survival curves for irradiated eukaryote cells.

Material and Methods: Mathematical simulation, algorithm, programming.

Results: Three models which take into consideration saturation of reparation systems in the irradiated cells at large doses were build and analyzed. The models differ in the number of reparable radiation damages they consider and whether they take into account formation of irreparable radiation lesions.

Conclusion: Mathematical models of the processes determining survival of irradiated cells which are based on the idea about saturation of repairing systems of irradiated cells at large doses and suggest limitation of the area of cellular chromatin served by one enzyme repairing complex were built. Qualitative correspondence of the calculated curves of survival and experimental data was shown. Possible perspectives of improvement of the mathematical models of the processes determining cell survival are discussed.

Key words: mathematical simulation, radiation lesion reparation, survival curves.

2004, vol 12, № 4, page 444

V. А. Potаpov, D. А. Hаshаchyh, S. V. Gеro

Influence of the age, lactation duration and breast volume on its electric conductivity

Annotation

Objective: To study the influence of age, breast (B) volume, lactation duration on breast electric conductivity in women determined with electromagnetic method to validate a new method of diagnosis.

Material and Methods: The study involved 52 women aged 14—60 with varions В volume and lactation duration without В pathology according to ultrasonography and mammography. The examination was done with electromagnetic (eddy current) method using a portable unit BC-5A.

Results: Dependence of В electric conductivity on the age, В volume, and lactation duration was revealed. It was established that В tissue electric conductivity increased with the age, by 35-40 years of age it reached its maximum, and decreased later. This was due to reduction in the water content of the tissues and reduced cell functional activity, which influenced polarization process in them. Electric conductivity increased with В enlargement due to increased polarization processes in the large tissue volume. Increase in lactation durution caused reduction in В functional capabilities, which resulted in electric conductivity reduction.

Conclusion: The study of В electric conductivity in women of various age with different В volume and lactation duration using electromagnetic (eddy current) method revealed dependence of these factors and the studied parameter. В electric conductivity reduces with the age and lactation duration and increases with В volume. These findings obtained using electomagnetic (eddy current) method are necessary for working out techiques of В disease diagnosis in women.

Key words: electric conductivity, breast, age, breast volume, lactation.

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