Przewlekła białaczka szpikowa jako drugi nowotwór mieloproliferacyjny u pacjentki z wieloletnim przebiegiem pierwotnej mielofibrozy

Chronic myeloproliferative neoplasms (MPN) can be categorised into Philadelphia-negative(Ph–) disorders and Philadelphia-positive (Ph+) chronic myeloid leukaemia (CML). Each ofthese neoplasms presents specific clinical and laboratory symptoms however occasionally thesediseases may coexist together, thus making classification difficult when such syndromes overlap.A case study is presented of just such an occurrence. This was a 44-year-old woman withprimary myelofibrosis (PMF) that had been recognised in 2004 according to obligatory WHOcriteria. After four years of follow-up the Ph chromosome appeared. Despite still having bonemarrow morphology characteristic of PMF, CML was found as shown by an atypical bonemarrow histology. A ten-month treatment with imatinib proved ineffective with complicationsarising of profound anaemia thereby requiring dose reduction, withdrawing the drug and redblood cell transfusion. It was only by administering nilotinib that the Ph+ clone was eradicated,however at the same time some abnormal Ph- clones were observed showing numerousmutations indicative of genetic instability. The clinical course and molecular biology resultsthen enabled recognition of two MPNs, namely PMF and CML, which had probably evolvedfrom two independent cell clones. This case therefore demonstrates that diagnosing MPN canbe challenging. The coexistence of two neoplasms should be taken into consideration in casespresenting an unusual clinical course and overlapping molecular abnormalities. Cytogeneticand molecular monitoring is thus important in such clinical cases. It allows not only a diagnosisMPN, but also new cell clones can be identified that lead to another MPN emerging as shownin this case report.Przewlekłe nowotwory mieloproliferacyjne (MPN) dzieli się na choroby bez obecności chromosomuFiladelfia (Ph) i przewlekłą białaczkę szpikową (CML) z obecnością chromosomu Ph.Chociaż każdy z tych nowotworów cechuje się określonymi objawami klinicznymi i laboratoryjnymi,to niekiedy choroby te mogą współistnieć, prezentując klinicznie i trudne do klasyfikacjizespoły nakładania. Przedstawiono przypadek 44-letniej pacjentki, u której w 2004 rokurozpoznano, zgodnie z obowiązującymi kryteriami Światowej Organizacji Zdrowia, pierwotnąmielofibrozę (PMF). Po 4 latach obserwacji u chorej pojawił się chromosom Ph, ale obrazmorfologiczny szpiku pozostawał taki, jak w PMF. Rozpoznano wówczas CML o nietypowejprezentacji histologicznej szpiku. Leczenie imatynibem, trwające 10 miesięcy, było nieskutecznei powikłane głęboką niedokrwistością, wymagającą zmniejszania dawki, odstawiania lekui przetaczania koncentratów krwinek czerwonych. Dopiero włączenie nilotynibu spowodowałoeradykację klonu Ph(+), ale jednocześnie ujawniło kilka nieprawidłowych klonów Ph(–)z licznymi mutacjami, świadczącymi o dużej niestabilności genetycznej. Przebieg klinicznyi wyniki badań molekularnych pozwoliły na rozpoznanie dwóch nowotworów PMF i CML,które najprawdopodobniej rozwinęły się z dwóch niezależnych klonów komórkowych. Prezentowanyprzypadek dowodzi, że diagnostyka MPN nie zawsze jest łatwa, a w sytuacjach o nietypowymprzebiegu klinicznym i nakładających się aberracjach molekularnych należy brać poduwagę współistnienie dwóch nowotworów. Duże znaczenie w takich przypadkach ma monitorowaniecytogenetyczne i molekularne, które nie tylko umożliwia dokonanie właściwego rozpoznaniaMPN, ale również, co przedstawiono w niniejszej pracy, pozwala na identyfikacjęnowych klonów komórkowych prowadzących do rozwoju kolejnego nowotworu

Secondary acute myeloid leukemia with monosomy 7 in Ph-negative cells in a patient with chronic myeloid leukemia in chronic phase

Cytogenetyczne zmiany klonalne w populacji komórek bez chromosomu Filadelfia (CCA/Ph–) są spotykane niezwykle rzadko w momencie rozpoznania przewlekłej białaczki szpikowej (CML). W przebiegu leczenia inhibitorami kinazy tyrozynowej (TKI) BCR-ABL1 stwierdza się je u 4–8% pacjentów. Zarówno patomechanizm i czynniki predysponujące ich wystąpienie, jak i znaczenie kliniczne nie są do końca jasne. Część z nich, w szczególności monosomia chromosomu 7, stanowi istotny czynnik ryzyka rozwoju zespołu mielodysplastycznego lub ostrej białaczki szpikowej (AML) o niekorzystnym rokowaniu. W pracy przestawiono przypadek 56-letniego chorego z CML, rozpoznaną w fazie przewlekłej w 2001 roku, początkowo leczonego hydroksykarbamidem, następnie interferonem alfa, a w dalszej kolejności trzema TKI, w tym imatynibem, nilotynibem i dazatynibem. W trakcie leczenia imatynibem u pacjenta stwierdzono monosomię chromosomu 7 w populacji komórek Ph–. Po 32 miesiącach od wykrycia CCA/Ph–, w trakcie leczenia dazatynibem, doszło do rozwoju wtórnej AML z obecnością złożonego kariotypu, przy współistniejącej CML w całkowitej remisji cytogenetycznej. Mimo zastosowanego leczenia, w tym chemioterapii indukującej opartej na daunorubicynie i arabinozydzie cytozyny (schemat 3 + 7) oraz jednego cyklu azacytydyny, nie uzyskano remisji AML. Pacjent zmarł z powodu powikłań infekcyjnych 4 miesiące po rozpoznaniu wtórnej białaczki. Opisany przypadek ilustruje złożony problem diagnostyczny i leczniczy wtórnej AML w trakcie długotrwałego leczenia CML. Hematologia 2011; 2, 3: 294–301Clonal cytogenetic abnormalities in Philadelphia chromosome-negative (CCA/Ph–) bone marrow cells are very rarely seen when chronic myeloid leukemia (CML) is diagnosed and have only been observed in 4–8% of patients undergoing treatment with BCR-ALB1 tyrosine kinase inhibitors (TKI). Moreover, the aetiology, clinical significance and potential risk factors of CCA/Ph– occurring remain unresolved. Some of these constitute significant risk factors, especially monosomy of chromosome 7, for the development of myelodysplastic syndrome or acute myeloid leukaemia (AML) associated with a poor prognosis. In this study the case of a 56-year old man in whom CML was diagnosed in 2001 is reported. Therapy was started with hydroxyurea followed by treatment with interferon alpha and TKI, including imatinib, nilotinib, and dasatinib. Monosomy of chromosome 7 in Ph– cells was detected during the imatinib therapy. The development of secondary AML, in the presence of a complex karyotype, occurred 32 months after CCA/Ph– was detected when a complete cytogenic response had been achieved during treatment with dasatinib. Despite receiving daunorubicine and arabinoside in a 3 + 7 induction regimen and one cycle of azacititdine, the patient responded poorly. Indeed, after 4 months from when AML had been diagnosed the patient died due to infectious complications. This case demonstrates the problems in making a diagnosis and of applying appropriate therapy during prolonged exposure to TKI in a patient with CML and a secondary AML. Hematologia 2011; 2, 3: 294–30

Genomic landscape of human erythroleukemia K562 cell line, as determined by next-generation sequencing and cytogenetics

We have performed detailed analysis of the genomic landscape of commercially available K562 cells, employing targeted enrichment of nearly 1300 cancer-related genes followed by next-generation sequencing (NGS) and also classical cytogenetics. Deep sequencing revealed 88 variants of potentially biological significance. Among them we have detected alterations in genes already known to be mutated in K562, such as TP53 but also in several other genes, which are implicated in tumorigenesis and drug resistance, such as MLH1, ASXL1 and BRCA1 as the most prominent examples. Fluorescence in situ hybridization (FISH) of interphases of K562 cells revealed multiplication of the BCR and ABL1 gene copies, as well as the amplification of the BCR-ABL1 fusion gene. Our results may help to better understand genomic instability of the blastic phase of CML represented by the K562 cell line and can help researchers who want to employ this cell line in various experimental settings

Excavations at Selib 2 in 2012

Selib 2 is a Meroitic settlement site of the 1st–4th century situated on the right bank of the Nile, 9 km upstream from the Christian pilgrimage site of Banganarti. Archaeological excavation in 2011/2012 was focused on two (of three recorded) Meroitic houses located in the northeastern part of the site. The mud-brick architecture was preserved at foundation level. Large amounts of pottery, stone finds and bone fragments were collected for further analysis

Possibilities of nanodiamonds application – biological and medical aspects

This topical review briefly discusses selected highlights of research on diamond nanoparticles obtained by different methods and their potential applications in biology and medicine. In recent years nanotechnology has aroused interest of large number of scientists who endeavor to obtain nanoparticles (which differ in size and structure of surface) using different methods, in order to determine their physical, chemical and biological properties that are in relation to the methods used in the process of their production. The knowledge developed in this way will be beneficial in an attempt to use nanoparticles more reasonably in various branches of science. The distinguishing features of carbon nanoparticles are their biocompatibility, photostability and easily chemically modified surface that result in high usefulness for intarcellular studies. What is more, low toxicity of nanoparticles with many cell lines is at the center of scientific interest. This, in turn, leads to a large number of biomedical applications. The property that nanodiamonds are able to penetrate into cells through endocytosis, allows to deliver the drug connected with nanoparticles into cancer cells. These features of nanoparticles have given many promising leads in nanooncology, in particular in drug delivery, diagnosis, imaging and therapy. This paper presents a summary of different classes of nanodiamond particles, the ways of their uptake into cells, an overview of the possible application of nanoparticles as nanocarriers and as a clinical theranostic platform, as well as advantages and disadvantages of using nanodiamonds in biomedicine

The Impact of O-Glycosylation on Cyanidin Interaction with RBCs and HMEC-1 Cells—Structure–Activity Relationships

With the aim of contributing to the knowledge about their potential therapeutic activity, we determined the biological activities of cyanidin and its selected O-glycosides in relation to erythrocytes (RBCs) and human dermal vascular endothelial cells (HMEC-1). Furthermore, on the basis of changes in the physical/functional properties of the cells, the structure–activity relationships of the compounds were determined. Concerning erythrocytes, we analyzed the antioxidant activity of the compounds and their impact on the RBCs’ shape and transmembrane potential. The compounds’ cytotoxic activity, ability to modulate apoptosis, cell cycle, and intracellular ROS generation, as well as inhibitory activity against AAPH-inducted oxidative stress, were determined in relation to HMEC-1 cells. We demonstrated that biological activity of cyanidin and its O-glycosides strongly depends on the number and type of sugar substituents, and varies depending on the extracellular environment and type of cells. The compounds are practically non-cytotoxic, and do not induce apoptosis or disturb the progression of the cell cycle. Additionally, the compounds alter the shape of RBCs, but they do not affect their transmembrane potential. They effectively protect erythrocytes against free radicals and affect intracellular reactive oxygen spices (ROS) generation under physiological and AAPH-induced oxidative stress conditions. Our results suggest a potential beneficial effect of cyanidin on the cardiovascular system

Protection of Erythrocytes and Microvascular Endothelial Cells against Oxidative Damage by <i>Fragaria vesca</i> L. and <i>Rubus idaeus</i> L. Leaves Extracts—The Mechanism of Action

The aim of this work is to determine the biological activity of ellagitannins rich extracts from leaves of raspberry (Rubus idaeus L.) and wild strawberry (Fragaria vesca L.) in relation to cells and cell membranes. Detailed qualitative and quantitative analysis of phenolic compounds of the extract was made using chromatographic methods. Cytotoxic and antioxidant activities of tested extracts in relation to erythrocytes and human vascular endothelial cells (HMEC-1) were determined by using fluorimetric and spectrophotometric methods. In order to establish the influence of the extracts on the physical properties of the membrane, such as osmotic resistance and erythrocytes shapes, mobility and/or hydration of polar heads and fluidity of hydrocarbon chains of membrane lipids, microscopic and spectroscopic methods were used. The results showed that the extracts are non-toxic for erythrocytes and HMEC-1 cells (up to concentration of 50 µg/mL), but they effectively protect cells and their membranes against oxidative damage. The increase in osmotic resistance of erythrocytes, formation of echinocytes and changes only in the polar part of the membrane caused by the extracts demonstrate their location mainly in the hydrophilic part of the membrane. The results indicate that tested extracts have high biological activities and may be potentially used in delaying the ageing process of organisms and prevention of many diseases, especially those associated with oxidative stress

Trudności w leczeniu chorego na ostrą białaczkę szpikową z mutacją FLT3-ITD i wysokim stosunkiem allelicznym — oporność na standardową chemioterapię indukującą w połączeniu z midostauryną

The development of targeted therapies in AML patients enabling treatment individualization, such as new FLT3 tyrosine kinase inhibitors, is a promising option for improving treatment outcomes and prolonging patient survival. However, the treatment of patients with a high FLT3-ITD allelic ratio (FLT3-ITDhigh) associated with an extremely unfavourable prognosis remains a major clinical problem. The study presents a case of a 20-year-old patient with FLT3-ITDhigh extramedullary AML at diagnosis. Individualized chemotherapy according to the DA ‘3 + 7’ regimen combined with midostaurin was administered. After the induction treatment, complete remission (CR) was not achieved. After second induction chemotherapy, CR1 was achieved with the presence of residua disease. One cycle of consolidation chemotherapy was then administered, and after myeloablative conditioning allogeneic hematopoietic stem cell transplantation from an unrelated donor was performed. The patient has remained in CR with no residual disease for 18 months.  Rozwój terapii celowanych u chorych na AML, umożliwiających indywidualizację leczenia, w tym zastosowanie nowych inhibitorów kinaz tyrozynowych FLT3, jest obiecującą perspektywą dla poprawy wyników leczenia i wydłużenia czasu przeżycia chorych. Jednakże problemem pozostaje leczenie chorych z obecnością wysokiego stosunku allelicznego FLT3 — FLT3-ITDhigh — związanego z wyjątkowo niekorzystnym rokowaniem. W pracy przedstawiono przypadek 20-letniego chorego na AML z obecnością FLT3-ITDhigh z lokalizacjami pozaszpikowymi AML przy rozpoznaniu. Leczenie chorego zindywidualizowano, stosując chemioterapię indukującą skojarzoną z midostauryną. Po zastosowaniu leczenia indukującego według schematu DA „3 + 7” nie osiągnięto u chorego całkowitej remisji (CR). Po włączeniu drugiej chemioterapii indukującej uzyskano CR z obecnością choroby resztkowej. Następnie podano jeden cykl chemioterapii konsolidującej oraz, po zastosowaniu kondycjonowania mieloablacyjnego, przeprowadzono przeszczepienie allogenicznych krwiotwórczych komórek macierzystych od dawcy niespokrewnionego. Pacjent pozostaje w CR bez obecności choroby resztkowej od 18 miesięcy