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Objective. The Philadelphia chromosome (Ph), due to t(9 ; 22)(q34 ; q 11.2), is the cytogenetic hallmark of chronic myeloid leukemia (CML) and is observed in more than 90% of the cases. Five to ten percent of patients with Ph positive CML have variant translocations involving chromosomes other than 9 and 22. Methods. We analysed 12 cases with variant Ph positive CML between 1994. and 2006. by conventional and molecular cytogenetic analysis. Conventional cytogenetic analysis was perfonned on bone marrow specimens after 24-48 h of culturing. Fluorescent in situ hibridization (FISH) was perfonned on slides of culture or freshly prepared slides. Results. Among the 164 patients with Ph 12 (7.2%) had variant translocations, involving one (n=11) or two (n=1) additional translocation partner chromosome. The median age was 46 years (range 18-69 years). Eight patients (66.6%) were in chronic phase, and four (33.3%) in accelerated phase, three with clonal evolution. The distribution of the breakpoints on sites other than 9q34 and 22q 11 has shown involvement of the following chromosomes: 6p23q23, 11q15 two patients, 12q24, 14p12, 15p13, 16p13, 17pl1, 18p12, 19p13, 21q22 and 22q13 two patients. Cytogenetic features of clonal evolution disease were combinations of double Ph. The BCR-ABL rearrangement were con finned by FISH. Conclusion. The clinical course and impact of these variants on long-tenn outcome is not well known, and conclusions are conflicting. Some studies have suggested that patients with variant Ph trans locations may have an adverse prognosis, while others have suggested that these translocationes have no impact on prognosis.

To evaluate chromosome aberration and fluorescent in situ hybridization (FISH) assays as a method to estimate of health risk, we monitored 9 male subjects occupationally exposed to low doses of both ionizing radiation and ultrasound during a period of over 3 years. Sampling was performed at 6-month intervals during a three-year period. First we used conventional chromosomal aberrations analysis. When the aberration frequency for a particular subject reached the background, we measured translocations in the final sample, using fluorescence in situ hybridization. Chromosome painting probes for chromosomes 1, 2, and 4 were used simultaneously. Dicentric and ring chromosomes were eliminated within a year. Translocations persisted and deviated from control values in all examinees. Translocations were detected long after unstable aberrations decreased to the background level. Fluorescence in situ hybridization-based translocation detection was a reliable method for monitoring chronic occupational clastogen exposure. Chromosome aberration assay correlated with translocation frequency. Stable chromosomal aberrations reflected cumulative genome damage during job exposure.

Aim of the study: The detection of chromosomal abnormalities characteristic of lymphomas, is important in the diagnostic workup of aggressive lymphomas given its impact on treatment strategies and prognosis. Recently this has been accomplished using FISH. In conformation with other methods for collecting samples makes the fine needle aspiration (FNA) attractive for diagnosis. Patients and Methods: We report the cytogenetic investigation in series of 59 patients with lymphoma (28 women and 31 men, median age 40, ranged 3-90 years), comprising 42 non-Hodgkin lymphomas (NHL) (32 abnormal) and 4 Hodgkin disease (HD) (2 abnormal)..

Over the last two decades the micronucleus assay has become an importante method in the estimation of genome damage in humans. The advantage of this method is its potential for detection of both clastogenic and aneugenic mechanisms. Micronuclei are mostly analysed in binucleated lymphocytes. The extension of micronuclesu count to mononuclear lymphocytes could provide new data for biomonitoirng purposes and therefore our study was done on both types of lymphocytes simultaneoulsy in subjects occupationally exposed to genotoxic agents. In order to evaluate this approach our study included 20 subjects exposed to gamma radiation and 25 subjects exposed to vinyl chloride monomer (VCM). The analysis was peroformed on 72 h lymphocyte cell culture with cytochalasin B as cytokinesis blocking agent. The mean value of micronucleus frequency of binucleated lymphocytes in subjects exposed to VCM was 7, 4% and in mononucleated lymphocytes 1, 1 %. In the population exposed to gamma radiation mena value in binucleated cells was 4, 6 % and in mononucleated lymphocytes 1, 4%. In both types of exposures micrnuclei frequency was significantly higher than in controls. The results show negative correlation between micronuclei frquency in monoclueated and binucleated lymphocytes for both agents. Although the model should be evaluated on a larger population, the observed results confirmed its value as additional information in risk assessment.

T-non-Hodgkin limfomi su izrazito maligne bolesti ; šire se vrlo brzo i ako se ne liječe bolesnici umiru. Sve do nedavno su rezultati liječenja bili loši, dok danas zahvaljujući vrlo agresinoj kemoterapiji ponekad kombiniranoj s radioterapijom i kirurškim zahvatom postiže se kompletna remisija u izrazito visokom postotku. Mogućnost izlječenja također je visoka od 85-90%. Materijal i metode. Na Zavodu za hematologiju i onkologiju Klinike za pedijatriju KBC Zagreb Referentnom centru Ministarstva zdravstva RH zs dječju hematologiju i onkologiju od 1990-2000 godine liječeno je 12 djece s T-non-Hodgkin limfomom (5 djevojčica i 7 dječaka ; prosječne dobi 7, 5 god) s protokolom NHL-BFM (vrlo sličnim protokolu za liječenje akutne limfatične leukemije). Svi su bolesnici ušli u 1. kompletnu remisiju u kojoj se i dalje nalaze. Zaključak: Premda je broj bolesnika relativno malen, postignuti rezultati liječenja vrlo su dobri i ne razlikuju se od rezultata koje postižu slični evropski centri za dječju hematologiju i onkologiju.

The aim was to improve the outcome of childhood acute lymphoblastic leukemia in Croatia. The achieved results are similar to the results achieved by other European Centres.

Na Zavodu za hematologiju i onkologiju Linike za pedijatriju KBC Zagreb referentnom ventru Ministarstva zdravstva RH za dječju hematologiju i onkologiju od 1990-2000 godine liječeno je 12 djece s T-NHL s protokolom NHL-BFM. Postignuti rezultati liječenja su vrlo dobri i ne razlikuju se od rezultata koje postižu slični europski centri za dječju hematologiju i onkologiju.

Alkalna fosfataza u leukocitima u tijeku terapije kronične granulocitne leukemije Glivecom

We detected an interstitial deletion of chromosome 2 [del(2)(p11.1p12)]in a lymphocyte culture of a patient. Karyotyping of the parents showed that the father had the same deletion. However, apart from this interstitial deletion he also had a small ring shaped Supernumerary Marker Chromosome(SMC), karyotype: 47, XY, del(2)(p11.1p12), + mar. FISH was performed on metaphase chromosomes using satellite DNA probe specific for chromosome 2 and whole chromosome painting specific probe wcp2. Hybridization was performed according to the protocol provided by the manufacturer. Other members of the family had normal karyotypes. FISH analyses revealed that the proximal break was right through the centromere of 2, spliting the chromosome 2 specific alpha satellite centromeric fragments into two smaller units, thus creating the functional centromere of the marker chromosome. The origin of marker choromosome was confirmed by FISH using wcp2: 47, XY, +mar. ish der(2)(wcp2+, D2Z1+). Marker chromosomes are structurally abnormal chromosomes in which no part can be identified. They represent a heterogeneous group of structural anomalies with different phenotypic expression depending on the size and genetic content of the marker and the level of mosaicism. Most of SMC have been studied using centromere specific DNA probes, which allow identification of chromosomal origin, and the chromosome specific libraries, which provide better insight into the structural composition and genetic content of the marker. The present case is a new case of supernumerary ring marker chromosome 2, identified by FISH, consisting of the proximal region of short arm of chromosome 2 and a part of its centromere.

We are presenting a family form of balanced translocation t(7 ; 14) found in mother and two sons. Mother had aortic cardiac murmur, without hemodinamical repercusions, children had almost identical clinical findings, significant supravalvular aortic stenosis, left ventricle intracavitary stenosis and multiple peripheral pulmonary stenosis but with no other clinical manifestatation of Williams-Beuren syndrome, except, perhaps, deep, metalic voice. The conventional chromosome analysis unexpectedly revealed a balanced translocation beetwen chromosomes 7 and 14, the same translocation was found in the mother and both of the children. Subsequent FISH analysis with WSCR probe showed that the break point has split the elastin region in proband, mother and both of children. The proband karyotype was interpreted, according to ISCN as 46, XY, t(7 ; 14) (q11.23 ; p12). ish t(7 ; 14) (D7Z1+, ELNsp ; D14Z1/D22Z1+, ELNsp+) mat, in other words, translocation had disrupted elastin region and may contribute to developmental defects in Williams-Beuren syndrome. Going through publications, we found that such a result was not yet published in genetic examination of supravalvular aortic stenosis and Williams- Beuren syndrome or some other conditions which can not be placed in any of these two terminal cathegories because of various phenotype characteristics.

The current diagnostics of haematological neoplasms along with morphological analysis, immunophenotyping and molecular analysis inevitably includes cytogenetic analysis. In this work the possibility of cytomorphological subclassification of haematological neoplasms from lymph node fine needle aspirates was examined without depending upon the referential histological diagnosis and cytogenetic analysis. In addition, the feasibility of cytogenetic analysis of the material obtained by lymph node fine needle aspiration (FNA) was examined. By analysing the findings of cytogenetic analysis and DNA image cytometry, it was decided to examine the possibility of comparing the findings and supplementing diagnostic possibilities of these methods. In 15 cases cytological diagnoses and cytogenetic analysis of haematological neoplasms were performed on the material obtained by lymph node FNA. In 12 of 15 cases histological diagnosis was made separately. A good cytohistological correlation was available in 9 of 12 cases (75%). Cytomorphological diagnoses in 10 of 15 cases (76%) were confirmed by the finding of a specific chromosomal translocation. In two cases cytological diagnosis did not correlate with the histological diagnosis and was confirmed only with specific chromosomal translocations. The lymphocytes obtained by lymph node FNA were adequate material for cytogenetic analysis - in 15 of 18 (83%) cases mitoses in cell cultures were obtained. In 13 of 15 (87%) cases clonal chromosomal abnormalities were detected, whereas in 2 of 15 (13%) cases a normal karyotype was found. DNA image cytometry was performed on nine samples, whereas in six samples the material was not sufficient. Although a small number of samples was analysed in the cases with identical cytomorphological diagnoses, the analysed histograms regarding the DNA index values showed heterogeneity. In conclusion, a cell culture sampled by FNA of lymph nodes is an adequate method for the chromosomal analysis. The specific cytogenetic abnormality associated with cytological diagnosis provides an opportunity to make a definitive diagnosis and provides a powerful approach when reference diagnosis on biopsy material cannot be obtained.

Cilj ove studije je bio potvrditi rezultate liječenja BFM-NHL studije te naći odgovarajuće mjere prevencije često po život opasnih situacija koje često karakteriziraju rani početak liječenja B-NHL bolesti u djece.Postignuti rezultati vrlo su dobri i bitno se ne razlikuju od zapadnoeuropskih centara.

U ovom radu iznose se rezultati liječenja nacionalne grupe za dječju hematologiju djece oboljele od akutne limfatičke leukemije u Hrvatskoj. Postignuti rezultati su gotovo isti kao i u vodećim europskim centrima za liječenje djece oboljele od akutne limfatičke leukemije.

The World Health Organisation classification of the hematologic neoplasms is based on morphology, immunophenotype, genetic analysis and clinical features. Aims: (1) To analyse the quality of the lymphocytic cell cultures which were sampled by the fine needle aspiration of the lymph nodes ; (2) to compare the cytogenetic abnormalities with the cytological diagnosis of neoplastic changes in the lymph nodes ; (3) to quantify the DNA content by the static DNA cytometry accompanied by the automatic picture analyser ; (4) to compare cytogenetic abnormalities with the changes in the DNA content measured by the static DNA cytometry. Methods: We have analysed 15 fine needle aspirates of the lymph nodes: 4 samples Burkitt lymphoma, 2 samples Non-Hodgkin peripheral T-cell type (2/2), 4 samples folicular cell lymphoma, I sample Non- Hodgkin lymphoma B-Iymphoblastic type, 2 samples acute myelogenus leukemia (M3 and M4) and I sample chronic myelogenus leukemia in dedifferentiation. The results have shown that the material was adequate in 85% samples. The cytogenetic abnormalities were analysed by the classical G- bending and FISH method. For the static DNA cytometry, the automatic picture analyser have been used comprising microscope, video camera and computer with SFORM software. Results: The normal karyotype was found in 33% (5/15) of analysed samples: Non-Hodgkin lymphoma peripheral T-cell type (2/2), Non-Hodgkin lymphoma B-lymphoblastic type (1/1) and Burkitt lymphoma (1/4). The specific chromosomal abnormalities common for the particular hematologic neoplasm which corresponded to the cytological diagnosis were found in 66% (10/15) of cases. The secondary complex cytogenetic abnormalities were found as well. In the five samples, 33% (5/15) cytogenetic abnormalities were not found. The results obtained by the static DNA cytometry have shown that the heterogeneity in the DNA amount exists. The> 30% cell percentage in the S + G2M phase or > 30% cell in the> 4 N indicates the rapid disease relapse, short survival or possibly correlates with complex chromosomal abnormalities. The cell culture from the fine needle aspirates of the lymph nodes has proved to be an adequate method for the chromosomal analysis that provides a powerful approach for diagnosing and subclassifying into subgroups with prognostic implication.

The study of chromosome translocations in relation with chromosome DNA-content after occupationla exposure to gamma radiation during non-destructive testing in industry. Eight subjects occupationally exposed to gamma radiation were analysed using fluorescent in situ hybridization (FISH) for chromosomes 1, 2 and 4. Distribution of translocations showed that chromosome 4 exhibited significantly the higher frequency of tranlsocations while chromosome 2 was significantly underrepresented respecting thier DNA content. Distribution of translocations in chromosome 1,2 and 4 are not in correlation with DNA content after occupational exposure to gamma radiation. Results of the study are in correlation with similar in vivo and in vitro studies. Non-random involvement of chromosomes point the necessity for standardization of chromosome sets in biodosimetry based on calculation of the genome equivalent

The conventional chromosome analysis of two brother and mother by high resolition banding unexpectedly revealed a balanced translocation between chromosome 7 and 14. Subsequent FISH analysis with WSCR and centromeric 7 probes showed that the break point has split the elastin region. The mother and her two sons carry the same translocation. The karyotype of brothers is: 46, XY, t(7 ; 14)(q11.23 ; p12).ish t(7, 14)(D7Z1, ELNsp)mat.

The rate of elimination, persistance and the accumulation of genetic damage are of great importance in epidemiologic studies of occupationally exposed population in industry as the conditions of induction of such damages are different from those described in medicine or nuclear plants. We investigated twenty subjects occupationally exposed to 192Ir or 137 Cs by chromosome aberrations assay and eight of them by fluorescent in situ hybridization (FISH)over a period of two years. Physical dosimetry showed that the annual doses kept below 50 mSv. Repeated analysis showed that the earlier found bicentric and ring chromosomes were eliminated during the period of one year, which confirms in the same time that detected aberrations were not constitutive. Our results show that frequency of translocations (0,022 translocations/cell on average) deviates from control values and that it does not correlated with years of employment.There is no possibility to make extrapolations or predict accumulation of trasnlocations by measuirng unstable aberrations as detected genome damage shows unequal distribution as a consequence of differences in individual activity at work place. In conclusion is that the evaluation relying on chromosome aberration assay is likely to underestimate the actual health risk of long term exposure to low doses of ionizing radiation in industrial radiography.

The development of new biomonitoring methods for evaluation of health risk due to environmental contamination or parental exposire redefines the child's position. The child is no longer a small human being but a unique entity with specific response to environment especially in first few years of life. Small blood sample and the quickness of the procedure, as it does not rquire cell culture, makes in vivo micronucleus assay elegant method for biomonitoring of children. The in vivo micronucleus assay is preferentially applied in genotoxicological experiments with rodents (Hayashi et al 1997). The results of biomonitoring of human population using this method are limited. We analysed 30 chiildren aged from 0-7 years. the results show that the average frequency of micronuclei in peripheral blood is 0,21%. There was no significant differences in micronuclei frequency between girls and boys. These agrees with published data for adult population (Xue et al., 1992). Small interindividual differences, low cost of method and quick results mark this method as suitable for biomonitoring of genome damage in children after exposure to clastogens and aneugens via environmental pollution. Small blood samples amkes the method applicable even in early postnatal period. Further research should give data on sensitivity of method and stability of this biomarker.

From NOvember20, 1996 to June 1, 2001 we have performed 906 molecular cytogenetic studies in 288 samples of peripheral blood, 49 amniotic fluids, 4 VCS, 4 chordocentesis and 10 tissues, 548 bone marrows and only 3 solid tumours. A variety of probes were used with numerous applications such as wcp, alpha satellite and specific region probes. Hybridised signals were scored under a fluorescence microscope using appropriate absorption and excitation filters. Microdeletion syndromes, rearrangements and marker chromosomes have been detected or chracterized by those techniques. Examples of chromosome markers include 29 small markers derived from chromosome 15, 22, 21, X and Y, a small marker 22 in case that also had a translocation (X;17). Several cases with "de novo" rearrangements were recognized. We found that an add(9)(p14;24) was a der(9)t(4;9) with monosomy 9p and partial trisomy 4p,der(13) was inversion duplication of 13(q22),cx translocation t(7;4;14) with insertion hence a partial trisomy of der 4, derivate chromosome 1 with centromere 15 on the terminal part of 1(p36) and that Elastin locus was disrupted by translocation t(7;14)(q11.23;p12). Molecular cytogenetic analysis of 12 workers exposed to the constant level of vinyl-chloride monomer were peroformed for detection of stable genome damage in order to estimate the genome risk. FISH studies in bone marrow and solid tumours characterised the origin of multiplechromosome rearrangements and helped in the interpretation of the diagnosis and/or prognosis. FISH was used for both, initial detection of specific abnormality with major prognostic and biologic impact and to the follow-up of treated patients by detection of MRD and identification of the origin of bone marrows cells following stem cell transplantation. I-FISH permits the evaluation of a large number of cells in detection of numerical changes and specific translocations. Hybridised signals were scored in about 100-700 interphase nuclei for each patient, with increased precision, since hundereds of cells(including nondividing cells) can be examined in a short time. Metaphase FISH was useful in detection of cryptic rearrangements and identification of marker chromosomes. The subtelomeres has been investigated with PNA probes in children leukemia. We concluded that the availability of new probes and the use of specialised techniques, such as FISH, M-FISH have improved significantlly the services offered by clinical cytogenetic laboratory in our hospital.

A subject occupationally exposed to 192Iridium during a period of 15 months who had shown a significant increase in chromosome aberrations with the annual dose below 50 mSv was analysed by in situ hybridisation in order to evaluate frequency of stable translocations and related health risk. Lymphocytes from the subject were scored using chromosome aberration assay and fluorescent in situ hybridization. Blood samples were taken on four occasions over a period of 2 years. The frequency of translocations measured by fluorescent in situ hybridization (0, 026 per cell) was significantly elevated over published control (0, 003 translocations per cell). Fluorescent in situ hybridization confirmed that chromosome damage was not the consequence of accidental overexposure it is concluded that the subject expresses radiation hypersensitivity.

From 1988 to 1998 one hundred forty eight patients with acute lymphoblastic leukemia (ALL) were enrolled into prospective study. The aim of the study was to evaluate efficacy of common treatment for acute lymphoblastic leukemia and to determine risk factor for outcome. Out of 148 patients, 31 (21%) had acute nondifferentiated leukemia (according to the FAB criteria) with expression of lymphoid markers. Those patients were treated in the same way as patients with ALL. According to the type of consolidation therapy patients were divided into three groups. Chemotherapy alone received 38 (38%) patients, and 30 (31%) patients received autologous stem cell transplantation (SCT) or allogeneic STC. There were no statistical difference in sex, age, FAB subtype, number of WBC and platelets in the time of diagnosis and incidence of cytogenetic abnormalities between these three groups. Median follow-up for chemotherapy group was 68 (range 12-98) months, for allogeneic SCT 102 (range 12-120) and for autologous SCT 99 (range 12-105) months. As a condition regimen the majority of patients treated with SCT received total body irradiation followed by cyclophosphamide. There was statistically significant difference in the source of stem cell for transplantation: in 20% of autologous SCT peripheral blood was used as source of stem cells. Relapse rate was significantly higher in patients receiving chemotherapy alone than in patients receiving either autologous or allogeneic SCT (83 vs. 45 vs. 27%, p

Na Zavodu za hematologiju i onkologiju Klinike za pedijatriju Šalata liječeno je 38 djece s akutnom limfatičkom leukemijom protokolom ALL-BFM 95 u periodu od 1.01.1995. do 31.12.1998.godine. Prva kompletna remisija postignuta je u 38 bolesnika (100% -33.dan ; u 32/84% -15.dan). Prvi relaps nastao je u 6 (15% ) bolenika (3 hematološki relaps, 1 meningelani relaps, 2 hematološki i meningealni relaps). 30.06.1999. u prvoj kompletnoj remisiji je i dalje 32 (85%) bolesnika ; 3 (8%) bolesnika je umrlo tijekom relapsa, a 3 je u drugoj remisiji. U grupi standardog rizika u prvoj kompletnij remisiji je 90%, u grupi sredjeg rizika 86%, a u grupi visokog rizika 40% bolesnika. Postignuti preliminarni rezultati potvrđuju vrijednost protokola ALL-BFM 95 u liječenju djece s akutnom limfatičkom leukemijom.

Hodgkinov limfom uz non-Hodgkin limfome nalazi se na trećem mjestu po učestalosti javljanja malignih bolesti u djece ; većina bolesnika danas, može se liječiti. Ukoliko dođe do relapsa što je rijetko, brzo se postiže druga, obično trajna remisija. Zadnjih 20-ak godina došlo je do značajnih promjena u strategiji liječenja Hodgkin limfoma: uloga radioterapije i kirurškog zahvata zančajno je smanjena dok dominantno mjesto zauzima citostatska terapija sa ili bez ( u dijela bolesnika koji uspješno odgovore na citostatsku terapiju) radioterapije. Danas se najveži napori uzažu u pronalaženje uspješne terapije praćene sa što manjim postotkom ranih i kasnih komplikacija liječenja specijalno pojave sekundarnog malignog oboljenja. U radu se iznose principi kao i rezultati suvremenog, vrlo uspješnog liječenja Hodgkin limfoma u djece.

Na Zavodu za hematologiju i onkologiju Klinike za pedijatriju Šalata liječeno je 38 djece s akutnom limfatičkom leukemijom protokolom ALL-BFM 95 u periodu od 1.1.1995. do 31.12.1998. Prva kompletna remisija postignuta je u 38 bolesnika (100%-33.dan ; u 32/84%-15.dan). Prvi relaps nastao je u 6(15%) bolesnika (3 hematološki relaps, 1 meningealni relaps, 2 hematološki i meningealni relaps). 30.6.1999. u prvoj kompletnoj remisiji je i dalje 32(85%) bolesnika ; 3(8%) bolesnika je umrlo tijekom relapsa, a w je u drugoj remisiji. U grupi standardnog rizika u prvoj kompletnoj remisiji je 90%, u grupi srednjeg rizika 86%, a u grupi visokog rizika 40% bolesnika. Postignuti preliminarni rezultati potvrđuju vrijednost protokola ALL-BFM 95 u liječenju djece s akutnom limfatičkom leukemijom.

Premda je imunofenotipizacija akutnih leukemija već dugo sastavni dio rutinskog dijagnostičkog postupka, još uvijek postoji neslaganje glede učestalosti i prognostičkog utjecaja imunofenotipskih kategorija, a posebno tzv. miješanih ili bifenotipskih akutnih leukemija (BAL). U ovom radu analizirana je učestalost citomorfoloških (FAB) i imunofenotipskih kategorija, kao i citogenetika 59 akutnih leukemija (AL) u djece liječene u Klinici za pedijatriju Šalata u razdoblju od kolovoza 1995. do prosinca 1998. godine. Od ukupno 59 AL, citomorfološka dijagnoza akutne limfatične leukemije (ALL) postavljena je u 42 (71%) bolesnika, akutne mijeloične leukemije (AML) u 16 (27%) bolesnika, dok su u jednog bolesnika nađene dvije leukemijske linije. Prema imunofenotipu, sve su AL razvrstane u tri skupine: AL bez koekspresije biljega - 40/59 (67.8%), AL s koekspresijom limfoidnih ili mijeloidnih biljega - 11/59 (18.6%), te bifenotipske AL (BAL) - 8/59 (13.6%). U skupini AL bez koekspresije biljega bilo je (prema FAB klasifikacji) 30 ALL i 10 AML, u skupini s koekspresijom biljega bilo je 8 ALL i 3 AML, dok je podjednak udio ALL i AML (4:3) nađen u skupini BAL (uz nalaz jedne biklonske AL). Pri tome, citogentski nalaz nije pokazao značajnu povezanost tzv. nepovoljnih promjena kariograma s imunofenotipski neuobičajenim kategorijama AL. Premda je u ovom istraživanju broj AL s koekspresijom biljega i BAL-a relativno malen u tijeku je dodatna obrada kliničkih parametara kako bi se ispitala njihova povezanost s laboratorijskim nalazima.

Here we report 4-year-old boy with per inv (9) mat who developed acute lymphoblastic leukemia, Ph+, after Langerhans cell histiocytosis. Stem cell disease may be responsible for such malignant manifestation in our case.

Uvod Dijagnostika akutnih leukemija u djece osniva se, kao i dijagnostika leukemija u odraslih, na FAB-klasifikaciji, imunološkoj tipizaciji te citogenetskoj analizi. Analizom naših rezultata željeli smo utvrditi korelaciju FAB subklasifikacije akutnih leukemija djece s imunološkim podtipovima te kromosomskim promjenama. Bolesnici i metode Analizirali smo 79 akutnih leukemija djece prema FAB-klasifikaciji, imunološkom fenotipu te citogenetskom nalazu. Bolesnici su bili mlađi od 15 godina. Rezultati U našoj analiziranoj skupini djece najčešće su nađene akutne limfatične leukemije (84,7%), dok su AML činile svega 15,3% akutnih leukemija u ovoj skupini. Od podtipova ALL najveći postotak činile su L2 leukemije (75,4%), L1 je nađena u 19,5% slučajeva, a najrjeđe se radilo o L3 subtipu. Od AML nađeni su svi podtipovi (od M1 do M8) osim podtipa M4. Koekspresija mijeloidnih biljega u citomorfološki dijagnosticiranih ALL nađena je u 14/57 (25 %) leukemija. U većini leukemija te skupine 12/14 (85%) dijagnosticiran je L2-podtip. U blastima svih leukemija citokemijski određena MPO bila je negativna. Ekspresija CD 34 antigena nađena je u visokom postotku i u L1 i u L2. Citogenetska analiza učinjena je u 57/78 (73,1%) obrađenih leukemija.U 9/57 (16 %) nalaz je bio uredan, u 42/57 (73 %) patološki, a u 6/57 (11 %) kultura nije uspjela. Zaključak U dječjoj dobi nalaze se svi oblici akutnih leukemija, samo što je ALL izrazito češća (AML:ALL=15,3%:84,7%). Od ALL u našem ispitivanju citomorfološki najčešće se nalazi podtip L2. U ALL našli smo visoku učestalost ekspresije mijeloidnog antigena u L2 podtipu uz imunofenotip "common". Korelacija morfoloških osobina blasta u ALL djece u usporedbi s imunološkim fenotipom pokazuje samo jasnu korelaciju morfološkog L3-podtipa i B-imunofenotipa zbog karakterističnih morfoloških osobina blasta u L3.

itogenetička analiza kromosoma pruganjem u stanicama koštane srži bolesnika sa akutnom mijeloičnom leukemijom otkrila je kompleksne promjene kariotipa. Mlađi muškarac sa obostranom limfadenopatijom vrata, splenomegalijom te povećanim medijastinumom imao je leukocitozu od 63.6 x 10/9/L sa 61% nezrelih stanica mijeloidne morfologije (FAB M-2) te trombocitopeniju. Koštana srž je bila infiltrirana istom staničnom populacijom Biopsija limfnog čvora pokazala je infiltraciju MPO+ blastima te nekrozu. Klasični pregled kromosoma dopunjen je hibridizacijom metafaza sa kromosom-specifičnim bojama (paints) za kromosome 1, 2, 4, i 13 (CytocellŽ) što je otkrilo dvije klonalne populacije: 45, xy, der(1)p(12)+6, del(6)(p21), -13, add(17)(p11)-20[25] i 46, idem, +der(?)t(1 ; ?)(q11 ; ?)[4]. Fenotipski (FACS) stanice KS/PK izražavaju mijeloidne biljege (MPO+, CD13+, HLA/DR+, CD34+, CD33+, CD15-, CD14-) uz 66% CD7+ blasta. Imunohistokemijski je obrađena ekspresija adhezijskih molekula L-selectina, ICAM te c-kit. Rijetki kariotipski nalaz u ovom mijeloidnom tumoru sa afinitetom za limfatična tkiva ukazuje na moguću povezanost izrazitih promjena u genetičkom materijala kormosoma 1 sa biološkim i kliničkim karakteristikama ovog tumora.

A younger male patient presented with acute leukemia of myeloid morphology (M2) with unusual clinical presentation of affection of lymphatic nodes, mediastinum and spleen. Immunophenotypicaly same antigen expression on blasts was found in BM, PB and biopsized lymph node. Cytogentics detected 46, xy, der (1) hsr(1 ; 1)(p11 ; ?)+6, del(6))p21=, -13, add(17p). Highly unusal and extensive involvment of chromosome 1 material in this complex karyotype suggest that some of the abberant genetic information related to Chromosome 1 in this patients might be responible for unusual hematological behaviour of the tumour

Opisana su tri genetska sindroma kod djece s akutnom limfoblastičnom leukemijom.

Blood lymphocytes of 12 radiographers working with Cs-137 source (37GBq) and 4 maintenance technicians were analysed by chromosome aberration assay and G-banding.The significant elevation of chromatid breaks confirms that the effect of mutagenic chemicals used in radiography should be considered as an additional parameter in final cytogenetic damage.

Umbilical cord blood is increasingly used as a source of hematopoietic progenitor cells for allotransplantation. Donor-derived buffy coat cells are considered optimal treatment for leukemia relapsles after transplantation of allogeneic bone marrow.

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We report on a month-old infant with dysmorphic face and several anomalies known to be associated with trisomy 13. Fluorescence in situ hybridization studies performed on metaphase cells allowed us to identify an extra material on the short arm of the chromosome 13 as a duplication of 13q22-qter

According to recent findings of DNA adducts formed after exposure to chemical mutagens it is suggested that certain parts of the DNA molecule, due to its structure, express higher fragility. In our study this speculation was investigated with well known mutagen and carcinogen, vinyl chloride (VCM). In a population occupationally exposed to VCM with elevated frequencies of chromosome aberrations the genome was analysed by G-banding. Clustering of chromosomes, chromatid braeks and deletions confirmed the non-random distribution of DNA damages. The most frequent damages were detected on bands: 1p13, 1q21, 1q23, 1q32, 3p21, 3q21, 5q31, 7p13, 9p21, 10q24, 11q23, 13q32. Very often the detected breakpoints coinceded with those detected in lymphoproliferative disorders and showed correlation with marked relative lymphocytosis detected in examinees. Methods of clinical cytogenetics and toxogenetics together offer potent tool in evaluating results that can be used for early detection of possible health risk and selecting biomarkers for biomonitoring of population exposed to known or suspected mutagens.

Children *86( with acute lymphatic leukemia ALL were treated with protocol ALL BFM 90 at the Centre for the Treatment of Leukaemia in Children, Department of Hematology and Oncology, Pediatric Clinic, {alata, University Hospital in yagreb, from 1st June 1992 to first June 1997. The results of treatment were analysed. The first complete remission was achieved in 86 100% / 33 day, in 71 83% / 15 daz. The first relapse occured in 12 14% patients / 9 hematologic relapse, 1 meningeal relapse, 2 hematologic et meningeal relapse. After a median observation time of 22 months, the overal probability for eventfree survival p/EFS is 81 =2%. 14 patients /16% died / 12 during relapse and 2 for other reasons. At this time p/EFS for each strategic group is 87 = 3% for SRG, 86 = 2% for MRG and 41 = 5% for HRG. These preliminary results indicate that for 89% of ALL patients risk adapted intensive therapy can provide a high chance for cure, Interestingly patients with more than 20% blasts in the bone marrow at day 15 suffer twice as many relapses as those with 5/20%, and four times more than those with less than 5% blasts.

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treatment of chronic myeloid leukemia with cord blood transplantion

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Acute promyelocytic leukemia - AML - M3...

Seven patients with acute promyelocytic leukemia (APL)treated with all-trans retinoic acid (ATRA) for remission (CR) induction and with chemotherapy (CT) for consolidation of CR underwent residual disease monitoring through reverse transcription polymerase chain reaction (PCR) for PML/retinoic acid receptor alpha (PML/RAR alpha) fusion transcript. In six patients PCR was positive immediately after ATRA induced CR and/or after the first CT course.Two patients became PCR negative following repeated courses of CT and they are in remission 7 and 14 months, respectively. Four other patients relapsed. This pilot study indicates that PCR for PML/RAR alpha is very sensitive assay for detection of residual disease in APL.

Aim. To evaluate the type and incidence of chromosomal abnormalities in 271 patients with acute leukemia and 71 patients with myelodysplasia. Methods. Banded chromosome analyses were performed on direct, 24- or 48-hour cultured and, in some cases, synchronized preparations of fresh bone marrow samples. Chromosome abnormalities were designated using the International System for Human Cytogenetic Nomenclature (ISCN, 1985). Results. Clonal chromosomal abnormalities were found in 119 (64%) patients with acute myelogenous leukemia. Fifty (75%) patients with acute lymphoblastic leukemia had clonal chromosomal abnormalities, half of which translocations. Clonal chromosomal abnormalities were found in 13 (57%) patients with acute unclassified leukemia and in 39 (55%) patients with myelodysplastic syndrome. Monosomies and deletions were predominantly revealed. Conclusions. Chromosomal abnormalities were found in 65% of patients with acute leukemia and in 55% with myelodysplasia. Most of them highly correlated with the specific F AB subtype of acute leukemia.

Prikaz sindroma i 2 obitelji koje su u cjelini citogenetski obrađene. Riječ je nakon Downova sindroma o najčešćoj genetičkoj bolesti povezanom s mentalnom retardacijom.