Thermal Effects on DNA Synthesis in Bone Marrow Cells of Patients with the Acute Leukemia.

Author(s):  
Hideki Kudo ◽  
Shinobu Sakamoto ◽  
Satoe Suzuki
Author(s):  
Валентина Викторовна Дмитриева ◽  
Николай Николаевич Тупицын ◽  
Евгений Валерьевич Поляков ◽  
Софья Сергеевна Денисюк

Применение методов и средств цифровой обработки изображений при распознавании типов клеток крови и костного мозга для повышения качества диагностики острых лейкозов является актуальной научно-технической задачей, отвечающей стратегии развития технологий искусственного интеллекта в медицине. В работе предложен подход к мультиклассификации клеток костного мозга при диагностике острых лейкозов и минимальной остаточной болезни. Для проведения экспериментальных исследований сформирована выборка из 3284 изображений клеток, представленных Лабораторией гемопоэза Национального медицинского исследовательского центра онкологии им. Н.Н. Блохина. Предложенный подход к мультиклассификации клеток костного мозга основан на бинарной модели классификации для каждого из исследуемых классов относительно остальных. В рассматриваемой работе бинарная классификация выполняется методом опорных векторов. Метод мультиклассификации был программно реализован с применением интерпретатора Python 3.6.9. Входными данными программы служат файлы формата *.csv с таблицами морфологических, цветовых, текстурных признаков для каждой из клеток используемой выборки. В выборке представлено девять типов клеток костного мозга. Выходными данными программы мультиклассификации являются значения точности классификации на тестовой выборке, которые отражают совпадение прогнозируемого класса клетки с фактическим (верифицированным) классом клетки. “Эксперимент показал следующие результаты: точность мультиклассификации рассматриваемых типов клеток в среднем составила: 87% на тестовом наборе, 88% на обучающем наборе данных. Проведенное исследование является предварительным. В дальнейшем планируется увеличить число классов клеток, объем выборок различных типов клеток и с уточнением результатов мультиклассификации The use of methods and means of digital image processing in the recognition of types of blood cells and bone marrow to improve the quality of diagnosis of acute leukemia is an urgent scientific and technical task that meets the strategy for the development of artificial intelligence technologies in medicine. The paper proposes an approach to the multiclassification of bone marrow cells in the diagnosis of acute leukemia and minimal residual disease. For experimental studies, a sample of 3284 images of cells was formed, submitted by the Hematopoiesis Laboratory of the National Medical Research Center of Oncology named after V.I. N.N. Blokhin. The proposed approach to the multiclassification of bone marrow cells is based on a binary classification model for each of the studied classes relative to the others. In the work under consideration, binary classification is performed by the support vector machine. The multiclassification method was implemented programmatically using the Python 3.6.9 interpreter. The input data of the program are * .csv files with tables of morphological, color, texture features for each of the cells of the sample used. The sample contains nine types of bone marrow cells. The output data of the multiclassification program are the classification accuracy values on the test sample, which reflect the coincidence of the predicted cell class with the actual (verified) cell class. “The experiment showed the following results: the accuracy of multiclassification of the considered types of cells on average was: 87% on the test set, 88% on the training data set. This study is preliminary. In the future, it is planned to increase the number of classes of cells, the volume of samples of various types of cells and with the refinement of the results of multiclassification


Author(s):  
Ольга Александровна Медведева ◽  
Святослав Николаевич Простаков ◽  
Николай Николаевич Тупицын ◽  
Александра Дмитриевна Палладина

В статье представлено описание факторов, влияющих на качество формирования базы эталонных изображений клеток костного мозга для диагностики острых лейкозов с применением методов компьютерной микроскопии. Отмечена важность контроля качества подготовки препаратов и микроскопа для применения в автоматизированных системах анализа изображений. Рассмотрены особенности регистрации цифровых микроскопических изображений клеток костного мозга в системах компьютерной микроскопии. Исследовано влияние фокусировки оптической системы микроскопа и уровня освещения препарата на формирование цифровых изображений клеток костного мозга. Установлены требования к условиям регистрации цифровых изображений, используемых в автоматизированных системах микроскопического анализа препаратов костного мозга. Предложена концептуальная модель базы эталонных изображений костного мозга, являющаяся основой для разработки инструментов эффективного распознавания клеток костного мозга в системах компьютерной микроскопии. Следование указанным требованиям к регистрации изображений призвано обеспечить надлежащее качество эталонной базы, что имеет непосредственной влияние на повышение точности и достоверности медицинской диагностики с применением методов компьютерной микроскопии. Результаты работы могут быть использованы в системах поддержки принятия врачебных решений при диагностике острых лейкозов The article describes the factors affecting the quality of the formation of a database of reference images of bone marrow cells for the diagnosis of acute leukemia using computer microscopy methods. The importance of quality control of specimen and microscope preparation for use in automated image analysis systems is noted. The features of registration of digital microscopic images of bone marrow cells in computer microscopy systems are considered. The effect of focusing of the optical system of the microscope and the level of illumination of the specimen on the formation of digital images of bone marrow cells is investigated. The requirements for the conditions of registration of digital images used in automated systems of microscopic analysis of bone marrow preparations have been established. A conceptual model of the base of reference images of bone marrow is proposed. It is the basis for the development of tools for effective recognition of bone marrow cells in computer microscopy systems. Following the specified requirements for image registration is designed to ensure the proper quality of the reference base of images, which has a direct impact on improving the accuracy and reliability of medical diagnostics using computer microscopy. The results of the work can be used in medical decision support systems for the diagnosis of acute leukemia


Blood ◽  
1980 ◽  
Vol 55 (2) ◽  
pp. 311-316
Author(s):  
R Lewensohn ◽  
U Ringborg

Unscheduled DNA synthesis (UDS) induced by melphalan, nitrogen mustard and ultra-violet irradiation was studied in bone marrow cells from myeloma patients. In a previous study, normal bone marrow cells in various stages of maturation were found to display a gradual decrease in UDS parallel with the process of maturation. Myeloma cells showed a similar pattern. Poorly differentiated myeloma cells exhibited a similar level of UDS to myeloblasts and erythroblasts. Irrespective of which repair-inducing agent was used, the relationship between the levels of UDS in the various cell types was constant. This indicates that the differences in the level of UDS in the various cell types was not due to differences in the uptake of the repair-inducing agent.


Blood ◽  
1979 ◽  
Vol 54 (6) ◽  
pp. 1320-1329
Author(s):  
R Lewensohn ◽  
U Ringborg

A technique has been developed for handling human bone marrow cells intended for the examination of DNA repair synthesis. DNA-repair synthesis, induced by melphalan and nitrogen mustard, was measured as the incorporation of 3H-thymidine, registered by autoradiography as unscheduled DNA synthesis (UDS). Comparison of various cell populations disclosed considerable differences in their UDS level, this generally being greatest for the blast populations. During maturation of both myelopoietic and erythropoietic cells, there was a decrease in the UDS level, which was lowest for the end-cell stage. The lymphocytes and monocytes differed considerably in their capacity for UDS. The developed technique would appear to offer an opportunity for determining the capacity for DNA-repair synthesis in malignant bone marrow cells, a factor that has been linked to sensitivity to alkylating agents.


Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1272-1272
Author(s):  
Jeff L. Yates ◽  
Hartmut Geiger ◽  
Gary Van Zant

Abstract DNA repair efficiency has been postulated to play a role in aging-associated phenotypes as well as in the generation of a variety of cancers. This is especially pertinent in highly proliferative tissues such as the lymphohematopoietic system, since the stem and progenitor compartments are responsible for maintaining proliferative demands within a restricted range for the lifetime of an individual. Hydroxyurea (HU) is a chemotherapeutic drug that targets DNA synthesis by inhibiting the synthesis of the nucleotide substrate resulting in stalled replication forks and single- and double-stranded breaks (DSBs) in the DNA. Recently, our lab has mapped a locus on mouse chromosome 7 that is involved in both organismal lifespan determination and HU sensitivity of bone marrow stem and progenitor cells in the HU-sensitive (25.9% killing), short-lived (540 days) DBA/2J (D2) and HU-insensitive (11.8% killing), long-lived (816 days) C57Bl/6J (B6) strains of mouse. To confirm that this locus is responsible for hydroxyurea sensitivity we generated congenic mice where the locus-containing interval was moved from B6 to D2 (D2. B6 chr. 7) and vice versa (B6. D2 chr. 7). When these animals were treated with HU it was found that the D2 locus imparts a high killing phenotype (38.0%) and the B6 locus confers a low killing phenotype (−4.2%). Using a flow cytometry-based in vivo Bromodeoxyuridine (BrdU) incorporation assay, we measured the recovery of DNA synthesis in the bone marrow in D2 and B6 mice after IP injection of HU (2mg/g). We first determined that DNA synthesis was completely inhibited within 15 minutes of injection and persisted for at least 3 hours in both mouse strains. At 4 hours, bone marrow cells of both strains began to incorporate BrdU, with B6 recovery more rapid than D2, 2.9+/−.5 vs. 7.9+/−3.9 percent BrdU positive cells (p=.01), respectively. Because HU has been used in the past to synchronize cells in G0/G1 and to measure cells in S phase, it was expected that BrdU incorporation would re-initiate within the G0/G1 compartment of cells. Indeed, bone marrow cells from D2 mice incorporated BrdU exclusively within the G0/G1 population. Surprisingly it was found that cells from B6 mice that had an S phase content of DNA prior to HU survived the insult and began to synthesize DNA. It was concluded that B6 bone marrow might have a more robust DNA damage response than that of D2. To study the DNA damage response in the bone marrow we treated mice with HU followed by BrdU and stained the bone marrow cells with an anti-BrdU antibody and an antibody to gamma-H2AX (gH2AX), a histone variant that becomes phosphorylated in the vicinity of DNA DSBs. In both D2 and B6 bone marrow cells it was shown that maximal gH2AX phosphorylation occurred within 1 hour and only occurred in the BrdU+ fraction of the bone marrow cells. Thus it can be concluded that HU causes DNA damage and these two strains of mouse differ in their response due in part to a locus on chromosome 7. Current studies are aimed at identifying the gene(s) of interest in the congenic interval, which include Tfpt and Prkcc.


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