Chromosomal abnormalities and DNA image cytometry of haematological neoplasms in fine needle aspirates of lymph nodes.

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.

Treatment of chronic myeloid leukemia in relapse after umbilical cord blood transplantation.

Umbilical cord blood (UCB) is increasingly used as a source of hematopoietic progenitor cells for allotransplantation. Donor-derived buffy coat cells are considered optimal treatment for leukemia relapses after transplantation of allogeneic bone marrow. Experience with relapses after UCB transplants are sparse. Here we report a girl who received an UCB transplant for chronic myeloid leukemia, relapsed after three years, failed to respond to donor buffy coat cells, but achieved a complete hematologic, cytogenetic, and molecular remission on interferon-alpha.

Partial trisomy 13 in an infant with a mild phenotype: application of fluorescence in situ hybridization in cytogenetic syndromes.

We report on a month-old infant with dysmorphic face and several anomalies known to be associated with trisomy 13. Fluorescence in situ hybridization (FISH) 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.

Risk estimates for balanced reciprocal translocation carriers--prenatal diagnosis experience.

An analysis was performed on 40 families at risk for an unbalanced rearrangement in the fetus because one of the parents is a reciprocal translocation carrier. The overall risk at second trimester prenatal diagnosis was 14% (8/57). The individual risk for unbalanced offspring at second trimester prenatal diagnoses and at birth were estimated using empirical data by Stengel-Rutkowski et al. (1988). The risks at birth ranged from 0%-21.6%. Most reciprocal translocations (22 or 55%) were at low risk. Without risk (7 or 17.5%), medium risk (6 or 15%) and high risk (5 or 12.5%) translocations were about equally represented and relatively infrequent. The analysis shows that the mode of ascertainment as well as the measurement of lengths of observed or probable imbalances cannot serve as a reliable risk predictor in individual counselling. In the translocations ascertained through spontaneous abortions the risk is frequently small or nonexistent, but remarkable exceptions to this rule are observed. Translocations discovered through unbalanced offspring were found to belong to different risk groups with the exception of the no risk group. Individual risk estimates have to be performed as a basis of genetic counselling before or during pregnancy so that parents with reciprocal translocations can make their choices regarding the available options.

Donor buffy-coat infusion and chemotherapy for leukemia in relapse after marrow transplantation.

A patient relapsing with blastic lymphoid transformation of chronic myeloid leukemia after bone marrow transplantation received donor buffy-coat infusion. Low-dose chemotherapy was added because of a rapid WBC increase. Complete hematologic and cytogenetic remission was obtained. The patient remained in complete hematologic and cytogenetic remission for four months until he died in an accident. Two patients with acute leukemia failed to respond to a similar treatment.

Molecular monitoring of minimal residual disease in acute promyelocytic leukemia by the polymerase chain reaction assay for the PML/RAR alpha (retinoic acid receptor-alpha) fusion transcript in patients treated with all-trans retinoic acid followed by chemotherapy.

Five acute promyelocytic leukemia (APL) patients who achieved a complete remission (CR) with all-trans retinoic acid (ATRA) underwent residual disease monitoring through reverse transcription polymerase chain reaction (PCR) for PML/retinoic acid receptor-alpha (PML/RAR alpha) fusion transcript. All received consolidation chemotherapy in CR, one in the form of autologous bone marrow transplantation (ABMT). In four of the patients PCR was positive for the PML/RAR alpha transcript immediately after ATRA treatment and/or after the first consolidation chemotherapy course. In the patient treated with ABMT, positivity was still detected six months after ABMT. One patient given five repeated courses of chemotherapy was PCR negative for PML/RAR alpha after 14 months in CR. Our pilot study confirmed that ATRA is a highly efficient induction therapy for APL in various stages of the disease, but ATRA alone cannot cure the disease. PCR should be considered a fundamental assay for assessing minimal residual disease in CR that will influence further treatment strategies and permit evaluation of treatment results.

Relationship between locations of chromosome breaks induced by vinyl chloride monomer and lymphocytosis.

The distribution of vinyl chloride monomer (VCM)-induced chromosome breaks was studied in cultured lymphocytes of subjects occupationally exposed to this gas. In the examined subjects, the mean group value of chromosome aberrations is 6.5% and for sister chromatid exchange (SCE) frequencies, the mean value per cell is 7.9. These values are significantly higher than in the control population. Occupational exposure to VCM caused lymphocytosis together with disturbances of mitogenic activity in lymphocytes stimulated by phytohaemagglutinin. The results of G-banding showed that sites of chromosome breakpoints caused by VCM can be related to the lymphatic tissue disorders.

[Trichorhinophalangeal syndrome]. / Triho-rino-falangalni sindrom.

A girl with tricho-rhino-phalangeal syndrome is described. Besides clinical and radiologic symptoms of type I of this syndrome (short stature, pear-shaped nose, prominent and elongated philtrum, small carious teeth, thin and sparse hair, badly shaped laterally protruding ears, deep voice, cone-shaped phalangeal epiphyses as well as shortening of metacarpal and metatarsal bones) the patient exhibits a group of abnormalities characteristic for type II (broad nasal bridge, thicker septum, laterally postured nostrils, heavier eyebrows, mild deafness and epilepsy). The girl also has the symptoms of primary hypothyroidism because of the ectopic thyroid gland, the first recorded instance up to now, so it is probably a coincidence.

Correlation of morphological FAB classification and immunophenotyping: value in recognition of morphological, cytochemical and immunological characteristics of mixed leukaemias.

Correlation between the FAB classification and immunophenotype was studied in 169 consecutive adult patients with acute leukaemia (AL). The lineage of leukaemic cells could be determined in the majority of cases, whereas 3 patients (1.8%) remained unclassified. In 22 out of 71 patients (31%) with acute myeloid leukaemia (AML) FAB M1 and M2 types, and in 5 out of 16 patients (31%) with chronic myeloid leukaemia (CML) in myeloid blast crisis, leukaemic cells did not express myeloid lineage-related markers, indicating asynchronous expression of cell markers in a substantial proportion of patients. Flow cytometric two-colour immunofluorescence revealed mixed AL immunophenotype in 6 out of 169 patients (3.4%). This group included five CD2+AML (5% of AML tested) and one undifferentiated AL expressing CD10(CALLA), CDw65(VIM-2). The former group included FAB M1, M2, M3 and M4 forms of AML with a single cell population, and an AML M2 patient with both cytochemically and immunologically two separate populations of leukaemic cells. This further illustrates the heterogeneity of the target cell(s) for leukaemogenesis and the level of differentiation of AML cells. However, there was no difference in the treatment response and the remission duration between AML patients and patients with mixed phenotype AML.

[The fragile X (Martin-Bell) syndrome]. / Fragilni X (Martin-Bellov) sindrom.

Fra (X) or Martin-Bell syndrome is the most common X-linked mental retardation with an incidence of 1/1000-2000 newborns. Chromatid break, double chromatid break or total loss of distal part of X chromosome (which occurs most often inside the C positive band q 27.3) is demonstrated in most male hemizygotes as mental retardation and specific phenotypic features. Fra (X) syndrome is proved in the cultured lymphocytes or fibroblasts with special cytogenetic methods. The prenatal diagnosis is possible by examining of amniotic fluid or the lymphocytes from the umbilical cord. We report two families with fra (X) syndrome. In the first one, 6 year- and 9-month-old boy with mental retardation and characteristic phenotypic features has been recognized as the carrier of fra (X) syndrome and after that his 4-year-old brother with similar symptoms. In the second family, there is a severe mentally retarded 3-year-old boy with fra (X) syndrome who besides typical phenotipic changes also exhibits symptoms of autism. The percentage of the cells with fra (X) chromosome in our patients (30%, 28%, 18%) is not correlated with the degree of their mental retardation. The mothers of our patients are the heterozygous carriers of the syndrome (3% and 1.5% fra (X) chromosome).

[Chronic autoimmune thyroiditis in patients with Turner's syndrome]. / Kronicni autoimuni tiroiditis u bolesnica s Turnerovim sindromom.

Studies concerning adult female patients with Turner's syndrome demonstrated that the impairment of the thyroid function is more often found in these patients than in the general female population. This is primarily related to a greater frequency of autoimmune thyroid diseases. Thyroid function was studied in a group of 23 female patients with Turner's syndrome, aged 7 to 24 years. Mean age was 15.6 years. Twelve patients had the karyotype 45X, three had mosaics 45X/46XX, one had the karyotype 45X/47XXX, while 7 had different structural X chromosome anomalies. They were all clinically euthyroid, except one which had plasma thyroxine (T4) values somewhat lower, and three which had plasma thyroid-stimulating hormone (TSH) concentrations somewhat higher than normal levels which could correspond to the so-called "subclinical" or "compensated" hypothyroidism. Elevated thyroid autoantibodies (TAA) were found in nine patients, whereas only two of these patients had goiter, simultaneously. It may perhaps be concluded that in female patients with Turner's syndrome, the atrophic form of autoimmune thyroiditis is more frequent than that associated with goiter (Hashimoto's thyroiditis), the latter being more prevalent in younger age groups in the general population. In view to the possible progression to hypothyreosis and the need for substitution therapy, the necessity of regular thyroid function follow-up especially in patients with positive TAA titers is emphasized. This is particularly important in female patients of younger age who are still in their growth and development period and in whom low thyroid hormone concentrations may lead to even greater growth retardation which is already compromised with the principal disease.

Four siblings with Robert's syndrome.

A Yugoslav family with one female and three male children with Robert's syndrome (RS) is described. To our knowledge, there is no other family with four siblings presenting this genopathy. Most of the symptoms of the fourth sibling were identical to those found in the SC syndrome, suggesting the same genetic origin. The same chromosomal changes which were discovered in the third sibling were also found in the fourth sibling. These chromosomal abnormalities could prove useful for antenatal diagnosis.