IGHV3-21 gene expression in patients with b-cell chronic lymphocytic leukemia in Ukraine

The aim of the study was to evaluate the frequency of IGHV3-21 gene usage and its clinical significance for patients with B-cell chronic lymphocytic leukemia (CLL) in Ukraine. Patients and Methods: Immunoglobulin variable heavy chain (IGHV) gene repertoire was studied in 189 CLL patients using reverse transcribed polymerase chain reaction and direct sequence of amplified products. Results: IGHV3-21 gene expression was found in 11 cases (5.8%), and its frequency was intermediate between Scandinavian (11.7%) and Mediterranean CLL (2.9%) cohorts. The most of cases (9 of 11) belonged to subset with heterogeneous HCDR3 (heteroHCDR3 subset), and only 2 cases – to subset with classical short ARDANGMDV motif (homHCDR3 subset). Six IGHV3-21 cases were mutated and 5 cases were unmutated. All unmutated cases (all were from heteroHCDR3 subset) had similarity of their HCDR3s with previously published sequences. The differences in overall (OS), progression-free (PFS) and treatment-free survival (TFS) for IGHV3-21 positive patients in comparison with CLL patients expressing the other IGHV genes were statistically insignificant. These survival parameters were comparable also for CLL patients with mutated IGHV3-21 gene usage and expression the others mutated IGHV genes. But remarkable feature of IGHV3-21 expressing patients was high incidence of solid tumors. They have developed in 4 IGHV3-21 positive cases (36.4%) and in 10 cases with expression of the others IGHV genes (5.6%, p = 0.0002). Furthermore, in small group of 6 patients with mutated IGHV3-21 gene expression, 3 patients had solid tumors and one underwent Richter transformation. Unmutated IGHV3-21 gene expressed patients had worse OS and PFS in comparison with CLL patients that expressed the others unmutated IGHV genes. Conclusion: Presented data are in agrement with the opinion about negative prognostic significance of IGHV3-21 gene expression regardless its mutation status. IGHV3-21 expression was associated with development of secondary solid tumors. Revealed high level of homology in heteroHDR3s subset might suggest about possible antigenic influence also, in addition to homHCDR3 subset that was proposed earlier.Цель: оценить частоту использования гена IGHV3-21 и его клиническое значение для больных B-клеточным хроническим лимфолейкозом (ХЛЛ) в Украине. Больные и методы исследования: репертуар генов вариабельных участков тяжелых цепей иммуноглобулинов (IGHV) изучали у 189 больных с ХЛЛ с помощью полимеразной цепной реакции на базе обратной транскрипции и прямого сиквенса амплифицированных продуктов. Результаты: экспрессия гена IGHV3-21 выявлена у 11 пациентов (5,8%), что занимает промежуточное положение между Скандинавской (11,7%) и Средиземноморской (2,9%) когортами. Большинство случаев (9 из 11) относились к подгруппе с гетерогенным третьим комплементарнымрегионом (heteroHCDR3 подгуппа) и только 2 случая — к подгруппе с коротким классическим ARDANGMDV мотивом (homHCDR уппа). есть IGHV3-21-позитивных случаев были мутированными и 5 — немутированными. Все немутированные случаи (все из heteroHCDR уппы) имели сходство HCDR3 с анее описанными последовательностями. Различия в общей выживаемости (OS), длительности периода до прогрессии заболевания (PFS) и начала лечения (TFS) для IGHV3-21-позитивных больных были статистически незначимы по сравнению с пациентами с ХЛЛ с экспрессией других IGHV-генов. Указанные параметры также сравнивали между больными ХЛЛ с экспрессией мутированных IGHV3-21- и других IGHV-генов. Отличительной чертой пациентов с экспрессией IGHV3-гена была высокая встречаемость солидных опухолей. Они развились в 4 IGHV3-21-позитивных случаях (36,4%) и в 10 случаях с экспрессией других IGHV-генов (5,6%, p = 0,0002). Кроме того, в небольшой группе больных (6) с экспрессией мутированного IGHV3-21-гена у 3 возникли солидные опухоли и 1 пациента — синдром Рихтера. У больных с экспрессией немутированного IGHV3-21-гена определяли худшие показатели OS и PFS по савнению с пациентами с экспрессией других немутированных IGHV-генов. Выводы: представленные данные согласуются с мнением о самостоятельном негативном прогностическом значении для больных с ХЛЛ экспрессии IGHV3-21-гена вне зависимости от его мутационного статуса. IGHV3-21-экспрессия была ассоциирована с развитием вторичных солидных опухолей. Выявленный высокий уровень гомологии в heteroHDR3s-подгруппе может свидетельствовать о возможном антигенном влиянии в дополнение к антигенному влиянию в homHCDR уппе, что было установлено ране

Новые публикации по судебной экспертизе

 This section presents translated abstracts of selected papers that appeared in the following periodicals: Forensic Science International  [www.elsevier.com/locate/forsciint], Forensic Science International: Digital Investigation [www.elsevier.com/locate/j.fsidi], Journal of Forensic Sciences [www.onlinelibrary.wiley.com] and Science & Justice [www.elsevier.com/locate/ scijus].  Представлены переводы рефератов избранных статей, опубликованных в зарубежных периодических изданиях: Forensic Science International [www.elsevier.com/locate/forsciint], Forensic Science International: Digital Investigation [www.elsevier.com/locate/j.fsidi], Journal of Forensic Sciences [www.onlinelibrary.wiley.com] и Science & Justice  [www.elsevier.com/locate/scijus].

РАДИОЛОГИЧЕСКИЕ И МЕДИЦИНСКИЕ ПОСЛЕДСТВИЯ ЧЕРНОБЫЛЬСКОЙ КАТАСТРОФЫ

From the position of a 25-years’ experience to overcome the health effects of Chernobyl the dynamics of the radiation environment, the first summarizing at the international level (1988), the results of completed research and practical monitoring are analyzed. Cohort of acute radiation syndrome (ARS) survivors under medical observation at the S.I. "Research Center for Radiation Medicine of the National Academy of Medical Sciences of Ukraine" is the largest. Within the 25 years the functional state of the major organs and body systems, and metabolic homeostasis for this category of persons were studied, a comprehensive assessment of their health, mental and physical performance were given, and risk factors and peculiarities of stochastic and non-stochastic pathology courses were identified, as well as a system of rehabilitation patients after ARS was developed. ARS survivors are suffering from chronic diseases of internal organs and systems (from 5-7 to 10-12 diagnoses at the same time). A correlation between acute radiation effects and specific HLA phenotypes were revealed. The dynamics of the immune system recovery after irradiation was studied. The role and prognostic value of telomere length and programmed cell death of lymphocytes in the formation of the cellular effects of ionizing radiation were determined for the first time. Differences between spontaneous and radiation-induced acute myeloid leukemias were found. Dose-dependent neuropsychiatric, neurophysiological, neuropsychological and neuroimaging deviations were identified after irradiation at doses above 0.3 Sv. It was shown that the lymphocytes of Chernobyl clean-up workers with doses 350 – 690 mGy can induce "the bystander effect" in the non-irradiated cells even after 19 years after exposure. The rates of cancer incidence and mortality of victims, the lessons and key problems to be solved in the third decade after the Chernobyl accident are considered.С позиций 25-летнего опыта преодоления медицинских последствий аварии на Чернобыльской АЭС проанализирована динамика радиационной обстановки, первые обобщения на международном уровне (1988 г.), итоги выполненных научных исследований и практического мониторинга. Когорта больных, перенесших ОЛБ и находящихся под диспансерным наблюдением в ГУ «Научный центр радиационной медицины НАМН Украины», в Украине остается самой многочисленной. За 25 лет изучено функциональное состояние основных органов и систем организма, метаболических процессов и гомеостаза у данной категории лиц; дана комплексная оценка состояния их здоровья, умственной и физической работоспособности, а также определены факторы риска развития и особенности течения стохастической и нестохастической патологии, разработана система реабилитации больных, перенесших острую лучевую болезнь (ОЛБ). Перенесшие ОЛБ и оставшиеся живыми страдают хроническими заболеваниями внутренних органов и систем (от 5–7 до 10–12 диагнозов одновременно). Выявлены коррелятивные связи между острым радиационным эффектом и определенными HLAфенотипами. Исследована динамика восстановления иммунной системы после облучения. Впервые установлены роль и прогностическое значение длины теломер и запрограммированной клеточной гибели лимфоцитов в формировании клеточных эффектов ионизирующего облучения. Найдены различия между спонтанными и радиационно-индуцированными острыми миелоидными лейкемиями. Дозозависимые нейропсихиатрические, нейрофизиологические, нейропсихологические и нейровизуализационные отклонения выявлены после облучения в дозах свыше 0,3 Зв. Показано, что лимфоциты участников ликвидации последствий аварии на ЧАЭС с дозами облучения 350–690 мГр способны индуцировать «эффект свидетеля» в необлученных клетках даже через 19 лет после воздействия радиации. Рассмотрены показатели онкологической заболеваемости, смертности пострадавших, уроки и основные проблемы для решения в третьем десятилетии после Чернобыльской катастрофы

RADIOLOGICAL AND MEDICAL CONSEQUENCES OF THE CHERNOBYL ACCIDENT

From the position of a 25-years’ experience to overcome the health effects of Chernobyl the dynamics of the radiation environment, the first summarizing at the international level (1988), the results of completed research and practical monitoring are analyzed. Cohort of acute radiation syndrome (ARS) survivors under medical observation at the S.I. "Research Center for Radiation Medicine of the National Academy of Medical Sciences of Ukraine" is the largest. Within the 25 years the functional state of the major organs and body systems, and metabolic homeostasis for this category of persons were studied, a comprehensive assessment of their health, mental and physical performance were given, and risk factors and peculiarities of stochastic and non-stochastic pathology courses were identified, as well as a system of rehabilitation patients after ARS was developed. ARS survivors are suffering from chronic diseases of internal organs and systems (from 5-7 to 10-12 diagnoses at the same time). A correlation between acute radiation effects and specific HLA phenotypes were revealed. The dynamics of the immune system recovery after irradiation was studied. The role and prognostic value of telomere length and programmed cell death of lymphocytes in the formation of the cellular effects of ionizing radiation were determined for the first time. Differences between spontaneous and radiation-induced acute myeloid leukemias were found. Dose-dependent neuropsychiatric, neurophysiological, neuropsychological and neuroimaging deviations were identified after irradiation at doses above 0.3 Sv. It was shown that the lymphocytes of Chernobyl clean-up workers with doses 350 – 690 mGy can induce "the bystander effect" in the non-irradiated cells even after 19 years after exposure. The rates of cancer incidence and mortality of victims, the lessons and key problems to be solved in the third decade after the Chernobyl accident are considered

Experimental Validation of a Methodology for Determining Soil pH and Specific Electrical Conductance in Samples of Geological and Soil Evidence in Forensic Environmental Investigations in the Absence of Standard Samples

The paper describes an experiment in validation of a forensic methodology for determining the hydrogen potential (pH) and specific conductance (SC) in samples of geological and soil evidence for the purposes of forensic environmental investigation. Validation was aimed at standardizing the conditions of aqueous extract preparation, since the conditions prescribed by corresponding regulations varied significantly. Given the absence of adequate standard samples at the time of the experiment, control samples had to be selected, and reference values of pH and SC in these control samples had to be determined through average values of the overall set of measurements. The experiment consisted of 5 operators independently conducting six parallel analyses of three control samples, each operator working at a different time and using their own assay kits. Both pH and SC values were measured in two different dilutions, each after 5 minutes, 1 hour, and 24 hours of holding time. Statistical calculations of the obtained set of results yielded reference values for target parameters in three control samples for different aqueous extract preparation conditions. It was demonstrated that when the extract is diluted to 1 : 2.5 soil/water ratio, measurements are on average 1.8 times higher for SC, and 0.20 pts lower for pH, compared to 1 : 5 soil/water dilution. Since 1 : 5 is the standard dilution for aqueous extracts, 1 : 2.5 dilutions call for a corresponding adjustment. Acceptable holding time between dilution and measurement has been established to vary between 5 minutes and 1 hour. When re-validated a year later, the obtained mean values of pH and SC in three control samples of soil fit within the uncertainty interval for adopted reference values.The outcomes demonstrate the stability of control sample properties and reliability of the applied methodology

Allelic Imbalances in Radiation—Associated Acute Myeloid Leukemia

Acute myeloid leukemia (AML) can develop as a secondary malignancy following radiotherapy, but also following low-dose environmental or occupational radiation exposure. Therapy-related AML frequently carries deletions of chromosome 5q and/or 7, but for low-dose exposure associated AML this has not been described. For the present study we performed genome-wide screens for loss-of-heterozygosity (LOH) in a set of 19 AML cases that developed after radiation-exposure following the Chernobyl accident. Using Affymetrix SNP arrays we found large regions of LOH in 16 of the cases. Eight cases (42%) demonstrated LOH at 5q and/or 7, which is a known marker of complex karyotypic changes and poor prognosis. We could show here for the first time that exposure to low-dose ionizing radiation induces AML with molecular alterations similar to those seen in therapy-related cases

Практика валидации методики определения pH и удельной электропроводности в объектах почвенно-геологического происхождения для производства судебно-экологической экспертизы при отсутствии стандартных образцов

The paper describes an experiment in validation of a forensic methodology for determining the hydrogen potential (pH) and specific conductance (SC) in samples of geological and soil evidence for the purposes of forensic environmental investigation. Validation was aimed at standardizing the conditions of aqueous extract preparation, since the conditions prescribed by corresponding regulations varied significantly. Given the absence of adequate standard samples at the time of the experiment, control samples had to be selected, and reference values of pH and SC in these control samples had to be determined through average values of the overall set of measurements. The experiment consisted of 5 operators independently conducting six parallel analyses of three control samples, each operator working at a different time and using their own assay kits. Both pH and SC values were measured in two different dilutions, each after 5 minutes, 1 hour, and 24 hours of holding time. Statistical calculations of the obtained set of results yielded reference values for target parameters in three control samples for different aqueous extract preparation conditions. It was demonstrated that when the extract is diluted to 1 : 2.5 soil/water ratio, measurements are on average 1.8 times higher for SC, and 0.20 pts lower for pH, compared to 1 : 5 soil/water dilution. Since 1 : 5 is the standard dilution for aqueous extracts, 1 : 2.5 dilutions call for a corresponding adjustment. Acceptable holding time between dilution and measurement has been established to vary between 5 minutes and 1 hour. When re-validated a year later, the obtained mean values of pH and SC in three control samples of soil fit within the uncertainty interval for adopted reference values.The outcomes demonstrate the stability of control sample properties and reliability of the applied methodology.Представлено описание эксперимента по валидации методики определения водородного показателя (pH) и удельной электропроводности (УЭП) в объектах почвенно-геологического происхождения для производства судебно-экологической экспертизы. Задачей валидации была унификация условий приготовления водной вытяжки из исследуемых объектов, поскольку в соответствующих регламентирующих нормативных документах условия существенно различались. В связи с отсутствием на текущий момент времени необходимых стандартных образцов возникла необходимость подбора контрольных образцов и оценки в них опорных значений pH и УЭП как средних значений совокупности результатов измерений. В эксперименте участвовали пять операторов, которые независимо друг от друга проводили по шесть параллельных анализов трех контрольных образцов, каждый оператор - в разное время, со своим набором реактивов и оборудования. Измерения pH и УЭП выполняли для двух разведений, каждое через 5 мин, 1 час и 24 часа. На основании статистических расчетов полученной совокупности результатов приняты опорные значения определяемых показателей в трех контрольных образцах при разных условиях приготовления водной вытяжки. Показано, что при разведении вытяжки в соотношении почва/вода 1 :2,5 в среднем результаты определения УЭП выше в 1,8 раза, а pH ниже на 0,20 ед. в сравнении с разведением почва/вода 1 : 5. Поскольку для водной вытяжки общепринятым является разведение 1 : 5, необходимо вводить поправку при разведении 1 : 2,5. В отношении времени выдерживания вытяжки до измерения установлено, что оно может варьироваться от 5 мин до 1 часа. При повторной валидации через год найденные средние значения pH и УЭП в трех контрольных образцах почвенных объектов находились в пределах интервала неопределенности принятых опорных значений. Полученные результаты свидетельствуют об устойчивости свойств контрольных образцов и надежности использованной методики

13q Deletions detected by fluorescence in situ hybridization for diagnosis and prognosis of chronic lymphoproliferative neoplasms

Aim. Determination of deletion of the long arm of chromosome 13 in the patients with chronic lymphocytic leukemia, diffuse large B-cell lymphoma and multiple myeloma to provide prognostic assessments of Chronic Lymphoproliferative Neoplasms (CLPN) sub-variants progression, and early detection of therapy resistant cases and relapses of CLPN. Methods. Preparations of bone marrow cells from all patients (n = 115) with CLPN were studied. Fluorescence in situ hybridization was performed using commercial test Vysis LSI D13S319 (13q14.3) Spectrum Orange/ Vysis LSI 13q34 Spectrum Green FISH probe kit (Abbott Molecular, USA). Results. The molecular cytogenetic investigations have revealed deletions of 13q in 38 % of the patients with CLPN. We also present a clinical case where the deletion of 13q is detected along with other cytogenetic aberrations that significantly impair a disease prognosis. Conclusion. The analysis of deletions of the long arm of chromosome 13 is an important diagnostic and prognostic criterion, which assists to optimizes the treatment of the patients with CLPN