Serum osteopontin and CD44 levels in lymphoreticular malignancies in children.

OBJECTIVE: Osteopontin (OPN) is an adhesive glycoprotein that interacts with a variety of cell surface receptors, including several integrins and CD44. OPN is expressed and secreted by numerous human malignancies. CD44 play an important role in tumor growth and metastasis. We aimed to evaluate serum levels of osteopontin and CD44 in patients with lymphorethicular malignancies in childhood. METHODS: We studied serum levels of CD44 and OPN levels of 54 patients (26, 18 and 10 patients with non-Hodgkin's lymphoma (NHL), Hodgkin's lymphoma (HL) and acute lymphoblastic leukemia (ALL), respectively) at the diagnosis. RESULTS: The mean levels of OPN were significantly higher in patients (5.42±8.24 ng/ml) than in controls (3.89 ±1.96 ng/ml). The mean levels of CD44 levels were also significantly higher in patients (3.82±2.31 ng/ml) than in controls (1.96±0.62 ng/ml), and significantly higher in the advanced stages than in early stages. The mean levels of the CD44 in NHL, HL and ALL were 3.49±2.00, 3.56±1.74, and 5.15±3.50 respectively. OPN and CD44 levels were found to be increased in parallel (p=0.003). A more advanced disease and/or poor prognostic factors were seen in 9 patients who had both serum CD44 and OPN levels higher than 2SD of the control. CONCLUSION: Elevated levels of both CD44 and OPN at the diagnosis may predict an unfavorable outcome in childhood leukemias and lymphomas (Tab. 2, Fig. 3, Ref. 44).

Unusually high frequency of a 69-bp deletion within the carboxy terminus of the LMP-1 oncogene of Epstein-Barr virus detected in Burkitt's lymphoma of Turkish children.

Burkitt's lymphoma (BL) in Turkish children is commonly associated with Epstein-Barr virus (EBV) infection. The C-terminus of the latent membrane protein 1 (LMP-1) of EBV is essential for transformation and the 30-bp deletion detected in this region has been implicated to be associated with a more aggressive malignant phenotype. To understand the molecular mechanisms underlying EBV pathogenesis in BL of Turkish children, we analyzed 30-bp deletion and 33-bp variable repeat regions of the LMP-1 gene from paraffin-embedded tumor tissues of 30 BL patients (mean age 5.9 years). Primer pairs spanning the 30-bp deletion and 33-bp repeat regions were designed for amplification by polymerase chain reaction (PCR). The PCR-amplified products were analyzed by gel electrophoresis, Southern blot hybridization, and DNA sequencing. Twenty-eight (93%) of 30 BL biopsy samples were EBV positive as determined by PCR. Variable copy numbers (ranging from 4.5 to 7) of the 33-bp repeat of LMP-1 gene were detected in these EBV-containing tumor samples. To determine the frequency of the 30-bp deletion of the LMP-1 gene, we sequenced the amplimers encompassing this region. Analyses of DNA sequence of 28 Turkish BLs have disclosed four patterns: the first (32% (9/28)) is identical to B95-8 with no deletion, the second (11% (3/28)) is identical to Asian NPC CAO strain with 30-bp deletion, the third (46% (13/28)) is prevalent in Turkish BLs with a longer deletion (69 bp), and the fourth (11% (3/28)) consists of a mixture of 30-bp and 69-bp deletion. The occurrence of high frequency of the 69-bp deletion appears to have no correlation with the disease site. Mutations found in the CAO strain were also detected in the Turkish BL clustering at the amino acids 322, 334, 338 and 342; whereas mutations specific for Turkish BL were clustered at amino acids 326, 352 and 361. To assess the EBV genotype with the changes in C-terminus of LMP-1 gene, we performed genotyping by PCR to differentiate type A and B strain. All 28 patients were infected by type A EBV. Such a high frequency of the larger size (69 bp) deletion has never been reported. Ascertaining the role of this deletion in BL pathogenesis will require further study.

Effect of nutrition on serum zinc concentration during pregnancy in Turkish women.

The serum zinc concentration in 101 pregnant women and 40 control subjects were determined by using atomic absorption spectrophotometer. The zinc levels were significantly low in 30 poorly nourished pregnant women compared to controls (P less than 0.01), whereas there was no statistical difference between the normal mean zinc values and that of well-nourished pregnant women (P greater than 0.05). This study revealed that nutrition is an important factor during pregnancy among Turkish women. In view of severe teratogenic effects of maternal zinc depletion in experimental animals as well as epidemiological evidence that maternal zinc deficiency could be a cause of severe congenital malformations of the central nervous system in humans, more extensive studies are definitely needed in Turkey.

The importance of cell size and surface marker analysis in childhood acute myeloblastic leukemia.

In order to evaluate the prognostic significance of cell size and surface marker expression, we evaluated 33 children with newly diagnosed acute myeloblastic leukemia by flow cytometry. We determined: the percentage of small, middle and large cells; large to small cell ratios (LS); large plus middle to small cell ratios (LMS); the percentage of surface markers expressed by each group of cell; the ratios of surface marker percentages expressed by the large blasts to that expressed by small blasts (LS for surface markers); and large plus middle blasts to that by small blasts (LMS for surface markers). For 'early prognosis', patients who could and could not achieve remission (n = 23 and 10) and for late prognosis, the patients who deceased or relapsed within the first 12 months of the treatment (n = 24) and who survived for more than 12 months (n = 9) were compared, in two classifications. CD3 percentages of the small cells of alive patients were significantly higher than that of dead or relapsed patients. LMS for CD3 and CD20 and LS for CD20 were higher in dead relapsed patients than that of alive patients. The total percentage of CD14 was significantly higher in dead relapsed patients than it was in the alive patients and CD3 was significantly higher in the group of patients who achieved remission than that of the patients who could not achieve remission. It was striking that, expression of CD3, CD7, CD22, CD33, CD14, CD15, CD34 increased or decreased as to cell size, whatever the prognosis. CD10, CD20 and CD13 were expressed on the large cells of the patients who could not achieve remission or died relapsed. We showed that, the blast cell size, individually does not have any prognostic significance in childhood AML and the prognostic significance of surface markers not only depends on their presence or absence but also on their relative configuration of expression by the blasts with different size.

Once daily ceftriaxone for meningococcemia and meningococcal meningitis.

Twenty children with meningococcal disease (15 with meningococcal meningitis and 5 with meningococcemia without meningitis) were treated with ceftriaxone, 80 to 100 mg/kg/day for 4 days. An additional 22 patients with meningococcal disease (13 with meningitis, 9 with meningococcemia without meningitis) were treated with penicillin G. On the basis of the Damrosch-Stiehm scoring system, 19 patients were classified in the poor prognostic group and were treated with antishock therapy. Clinical recovery time and normalization of CSF were compared in two groups. When the complications were compared, necrotic skin lesions were more frequently seen in the penicillin G group than in those who received ceftriaxone. Ceftriaxone is an effective and safe drug and offers the advantage of once daily administration for treatment of meningococcal disease in pediatric patients.

Soluble L-selectin increases in the cerebrospinal fluid prior to meningeal involvement in children with acute lymphoblastic leukemia.

Soluble L-selectin was determined in the CSF samples of 20 children with CNS leukemia at the time they had blasts in CSF and/or clinical findings of CNS involvement; 17 CSF fluid samples were obtained from 17 of these 20 children, 29-91 days before the appearance of CSF cytological and/or clinical findings of CNS involvement; while 15 CSF samples were withdrawn from among the same group of children, after treatment of meningeal leukemia. In addition, CSF sL-selectin was also assayed in 17 children with ALL, who remained in complete remission at least for a year and, as controls, in 12 children without malignant or meningeal disorders. There was no significant difference in CSF sL-selectin levels between the children with ALL without evidence of meningeal involvement and the controls (1.34 +/- 0.21 ng/ml, 1.46 +/- 0.18 ng/ml respectively, p > 0.05). However, in children with CNS leukemia, not only at the time CNS involvement was diagnosed, but also 29-91 days before the diagnosis of CNS leukemia, the concentrations of the CSF sL-selectin (12.41 +/- 2.14 ng/ml, 7.70 +/- 1.60 ng/ml respectively) were significantly higher than those in controls (p < 0.001 and p < 0.01 respectively). After treatment and disappearance of the blasts in CSF, sL-selectin was found to be decreased and even normalized in the majority of children who had meningeal involvement (2.87 +/- 2.14 ng/ml). In 5 children, the CSF sL-selectin remained high, after the blasts in CSF had disappeared and CNS leukemia recurred within 3 months in 4 of these 5 children. In conclusion, assay of sL-selectin in CSF seems to be a good diagnostic tool in the detection of CNS involvement in children with ALL. This method may also be used as an indicator, in prediction of the CNS leukemia, which is going to develop.

High prevelance of chronic magnesium deficiency in T cell lymphoblastic leukemia and chronic zinc deficiency in children with acute lymphoblastic leukemia and malignant lymphoma.

Magnesium and zinc are the elements having essential roles in regulation of cell growth, division and differentiation. There have been some studies in the literature suggesting an association between the deficiency of these elements and the development of malignant disorders. In this study hair and serum zinc and magnesium levels were investigated in children with acute lymphoblastic leukemia (ALL) and malignant lymphoma (ML) at the time of initial diagnosis. Ten children with T-cell ALL, 10 children with B-precursor ALL, 5 children with Burkitt's Lymphoma (BL), 11 children with Hodgkin's lymphoma (HL), 10 children with non-Burkitt non-Hodgkin's lymphoma (NBNHL) and 12 age and sex matched healthy children as a control group were included in the study. Mean hair magnesium levels in all of the groups of the patients were lower than the levels in the control group but the difference was statistically significant only in the children with T cell ALL comparable to the controls (28.9+/-3.9 microg/g and 87.6+/-18.5 microg/g respectiveley, p<0,05). Mean serum magnesium levels in all the cohorts were not significantly different than those in controls (p>0.05 in each comparison). Mean hair zinc levels in the patients with T-cell, B-precursor ALL, BL, HL, NBNHL were 103.4+/-14.6 microg/g, 100.9+/-7.8 microg/g, 91.1+/-19 microg/g, 72.5+/-9.1 microg/g, 103.2+/-12.2 microg/g respectively. Each of these levels were significantly lower than the mean hair zinc levels of the control group (141.2+/-9.6 microg/g, p<0.05 in each comparison). Although mean serum zinc levels in all of the groups were also decreased, the differences were statistically significant only in the groups with B-precursor ALL, HL and NBNHL (75.9+/-5.29 microg/dl, 68.6+/-7.3 microg/dl, 85.7+/-5.5 microg/dl respectively) when compared with controls (105.1+/-9.9 microg/dl, p<0.05 in each comparison). Hair magnesium and zinc levels showed a positive correlation with each other in all the groups (r congruent with 0.5). No significant difference was found in the mean hair/serum magnesium and zinc levels between malnourished and nonmalnourished patients. In conclusion, regarding the results of our study and previous data in the literature chronic magnesium and zinc deficiency seems to be associated with the development of ALL and malignant lymphoma in a group of patients.

Coexistence of life threatening chemotherapy related leukoencephalopathy, saggital sinus thrombosis and multiple organ failure in a child with acute lymphoblastic leukemia: an unusual case with clinical recovery.

A nine-year old girl with T cell acute lymphoblastic leukemia (ALL) had acute severe neurologic complications at the end of the remission-induction chemotherapy course. Thirty-six hours following triple intrathecal (IT) therapy and intravenous (IV) administration of L-asparaginase (L-asp), tetraplegia developed and she became unconscious. She had bouts of hypertension and persistent tachycardia unresponsive to digitalis therapy. Magnetic resonance imaging (MRI) showed multiple brain white matter hyperintensities and filling defects in the saggital sinus, suggesting thrombosis. Over the 40 days, in addition to her neurologic compromise she also had transient diabetes mellitus, severe hyperlipidemia, hypoproteinemia and edema, liver and heart failure and staphylococcus aureus sepsis with prolonged bone marrow depression. Despite, coexistence of all these chemotherapy related complications, her neurologic functions and multiple organ failure improved gradually. After a 70 days' period of interruption, chemotherapy was resumed and continued without any further complications. Although, the etiology of her extensive sensitivity to some drugs remains unclear, we believe that it is important to document these unusual events in this child.

Burkitt's lymphoma in Turkish children: clinical, viral [EBV] and molecular studies.

Eighty-one Turkish children with Burkitt's lymphoma (BL) were observed during a period of 24 years (1968-1992). The diagnosis was established histologically according to WHO criteria. BL represented 48.5% of NHL in this series. The median age of patients was 5 years with a sex (M/F) ratio of 2.3/1. The most common primary site of tumor involvement at initial presentation was the abdomen (70.4%), which was followed by facial tumors, in particular the jaw and orbit (45.7%). The majority of the patients (84.0%) were in advanced stages (C and D) at initial diagnosis. Facial tumors observed in Turkish children with BL were more similar to African Burkitt's lymphoma than American or European cases. High titers of antibodies against VCA and EA of EBV were also observed in 32 recent cases of BL. Preliminary molecular and immunologic studies revealed EBV-DNA (type I) and T cell deficiency. The clinical presentation, median age, and association with EBV revealed that BL appears to be inbetween African and non-African types in Turkish children. This will be further elucidated in the future by direct examination of tumor cells for EBV and investigation of the molecular characteristics in these cases.

Erythroblastopenia and leukopenia in the patient with severe herpes zoster treated with intravenous acyclovir.

A seven-year-old girl with transient leukopenia and erythroblastopenia which developed after the administration of acyclovir is presented. Acyclovir infusion was given in a dose of 5 mg/kg/every eight hours. On the third day of therapy the hemoglobin level fell to 9.9 g/dl, the hematocrit was 26%, the white blood cell count 4000 percent/mm3, red blood cells 3.2 million percent/mm3. Bone marrow aspiration showed a decrease in the number of erythoblasts and a relative increase in the number of promyelocytes and myelocytes. Therapy was discontinued on the fifth day, and on the seventh day the findings were normal including the bone marrow aspiration. We could not find any other reason which would cause transient erythroblastopenia and leukopenia in our patient.

Relationship between high leukocyte count and cell size in childhood acute myeloblastic leukemia.

In order to determine the significance of cell size together with high leukocyte count (>30x10(9)/L) in acute myeloblastic leukemia (AML), we evaluated the percentages of small, medium and large cells in 33 children with AML. All of the 10 patients with a high leukocyte count and 14 of the 23 patients with a low leukocyte count (<30x10(9)/L) died or experienced a relapse within the first year. The mean small cell percentage of patients with high leukocyte counts was significantly lower than that of patients with low leukocyte counts (p<0.05). The percentages of small, medium and large cells of patients with high leukocyte counts and of patients with low leukocyte counts who died or experienced a relapse within the first year were similar. The percentage of medium cells of patients with high leukocyte counts was significantly higher than that of surviving patients with low leukocyte counts (p<0.05). The mean percentages of small, medium and large cells were similar in patients who died or experienced a relapse and surviving patients with low leukocyte count. We conclude that cell size has prognostic significance when the leukocyte count at admission is over 30x10(9)/L, although confirmation seems necessary with a larger population of patients.

Biphenotypic characteristics, cell size and prognosis in childhood acute myeloblastic leukemia.

In order to determine the prognostic significance of cell size together with expression of biphenotypic markers in childhood acute myeloblastic leukemia (AML), we evaluated the cell size of children with AML, 12 with and 21 without biphenotypic markers. The patients were followed up for at least 12 months. The cells which were stained with FITC conjugated surface marker antibodies were divided into small, middle or large cell groups according to their mean channel number of forward scatter by flow cytometry. Nine of 12 biphenotypic and 15 of 21 non-biphenotypic children either died or relapsed within the first 12 months. The percentages of the small, middle and large cells were similar in children and in deceased patients, regardless of whether or not they expressed biphenotypic markers. We believe that biphenotypic marker expression is a poor prognostic factor regardless of cell size.

Aggressive Kaposi's sarcoma in children: a case report.

A three-year-old-boy with generalized Kaposi's sarcoma (KS) is presented. The child died of progressive pulmonary insufficiency on the eighteenth day of the course of his illness, the tenth hospital day. On postmortem examination diffuse KS infiltration was observed in the respiratory and gastrointestinal tracts, lymph nodes, liver, spleen and thymus. The patient was considered to be a case of KS unrelated to AIDS because of his negative HTLV-III antibody and epidemiologic characteristics, and therefore was believed to have primary aggressive KS.

Production of granulocyte colony stimulating factor, granulocyte macrophage colony stimulating factor, and tumor necrosis factor alpha during remission and infections in patients with acute leukemia.

We measured circulating serum levels of granulocyte colony stimulating factor (G-CSF), granulocyte macrophage colony stimulating comparable to the levels of these factors in 12 children with acute febrile infections without malignancy or hematological disorders and 15 age matched healthy controls. There were significantly elevated levels of G-CSF, GM-CSF and TNF alpha, in 12 children with infections without leukemia, as compared with controls. Also in 18 leukemic children with infections serum G-CSF and TNF alpha levels were significantly higher than those in the leukemic children without infection and healthy controls, whereas no significant difference was noted in the GM-CSF levels in these groups. Although elevation in TNF alpha levels in response to infections were similar in the children with and without leukemia, in the G-CSF levels lower elevation was noted in the leukemic children with infections as compared to the children with infections without leukemia. Despite leukopenia enhanced the production of G-CSF, even in leukopenic children with leukemia and infections, serum G-CSF levels were still lower than those for the children with infections without leukemia. We concluded that, the production of G-CSF and GM-CSF as a response to infection was deficient in the patients with acute leukemia in remission, probably due to the maintenance and reinforcement chemotherapy. Therefore, the use of recombinant G-CSF may be recommended in the infections of these patients.

Zinc status and cellular immunity in pediatric Hodgkin's disease.

In the present study we investigated serum, plasma, RBC and hair zinc concentrations in pediatric Hodgkin's cases at initial presentation and in remission (measured by atomic absorption spectrophotometry). In addition, immune parameters such as total lymphocyte counts (TLC), E-rosette test (E-R), lymphocyte proliferative (LP) responses to PHA, and skin tests to four antigens were simultaneously investigated in some cases. Although all zinc levels were found to be significantly low before treatment, suggesting chronic zinc deficiency, serum and plasma zinc levels returned to normal during remission. A positive correlation could be detected only between serum zinc levels and lymphocyte blastogenic response to PHA. Hodgkin's patients with complete anergia also showed significantly lower serum zinc levels and LP responses to PHA than the patients without anergia.

Myocardial performance index combining systolic and diastolic myocardial performance in doxorubicin-treated patients and its correlation to conventional echo/Doppler indices.

This study was designed to evaluate the utility of myocardial performance index (MPI) in anthracycline cardiotoxicity. The MPI measures the ratio of total time spent in isovolumic activity (isovolumetric contraction time and isovolumetric relaxation time) to the ejection time, thus giving a global index combining systolic and diastolic myocardial performance. In this study, MPI was measured in 35 doxorubicin-treated children (aged 108.5+/-55.31 months, 23 males and 12 females) in sinus rhythm and 32 age-matched controls, and it was compared with conventional Doppler echocardiographic parameters. The isovolumetric contraction time was prolonged (38.37+/-24.43 vs 26.37+/-15.53, p <0.02) and ejection time was shortened (231.91 +/- 28.87 vs 256.21+/-19.55, p<0.001) in doxorubicin-treated patients compared to that in normal children. The isovolumetric relaxation time did not show significant difference between patients and control group (60.11+/-10.92 vs 61.06+/-12.12, p>0.05). MPI was significantly increased in doxorubicin-treated patients compared with that in control groups (0.42+/-0.07 vs 0.34+/-0.06, p<0.001), and significant correlation was observed between MPI and fractional shortening, ejection fraction, and left ventricular end diastolic and end systolic diameters (respectively, r = -0.508, p <0.002; r = -0.532, p<0.001; r = 0.467 p<0.005; r=0.606, p<0.001). Also, a weak correlation was found between MPI and duration of the disease and patient ages (r = 0.393, p < 0.02; r = 0.379; p < 0.02). However, there was no correlation between MPI and cumulative doxorubicin dose (r = 0.311, p > 0.05) and diastolic Doppler parameters in doxorubicin-treated patients. We think that MPI may be a useful parameter in monitoring left ventricular dysfunction in anthracyline-treated patients.

Zinc and anergy in pediatric Hodgkin's disease in Turkey.

Blood (serum, erythrocytes) and hair zinc levels were determined in 60 biopsy-proven pediatric Hodgkin's disease cases at diagnosis. Cellular immunity also was assessed through total lymphocyte counts, E-rosette formation, lymphoproliferative response (LP), and delayed cutaneous hypersensitivity tests to dinitrochlorobenzene, streptokinase-streptodornase, purified protein derivative and phytohemagglutinin (PHA) in some of these patients. Interestingly, anergic patients unresponsive to four antigens showed significantly more depressed serum zinc levels as well as decreased lymphoproliferative response to mitogen (PHA). A positive correlation could be shown between serum zinc level, cutaneous anergy and LP. A possible contributing role of zinc deficiency in defective cell mediated immunity in Hodgkin's disease was proposed, and administration of oral zinc, as a natural immunostimulant is considered in this lymphoma.

High risk subgroup of acute myelomonocytic leukemia (AMML) with orbito-ocular granulocytic sarcoma (OOGS) in Turkish children. Retrospective analysis of clinical, hematological, ultrastructural and therapeutical findings of thirty-three OOGS.

Thirty-three patients presenting with orbito-ocular granulocytic sarcoma (OOGS) and acute myelomonocytic leukemia (AMML) were diagnosed in Turkish children from 1963 to 1983. OOGS, characterized by exophthalmos, chemosis and orbital masses, was observed in 33 (27.2%) of 121 AML patients compared with 41 children of AMML without ophthalmic tumors during the same period. Eye tumor and bone marrow aspirates were also studied under light and electron microscopies. The comparison of the hematological parameters did not indicate any statistical difference between the groups. Despite similar chemotherapy regimens administered to all patients, the mean survival time was 8.7 months in the OOGS group, which is significantly shorter compared to those without OOGS (28.6 months) (p less than 0.01). These cases may be classified as a "high risk" subgroup of childhood AMML.

Hodgkin's disease in 82 Turkish children diagnosed over a 10-year period: epidemiological, clinical, and histopathologic features and prognosis with prolonged chemotherapy.

In this study, 82 Turkish children with Hodgkin's disease (HD) between 1 and 14 years of age and diagnosed over a 10-year period were evaluated retrospectively. More than half of the patients (54%) presented with advanced stages of HD. Mixed cellularity (MC) was the most frequent (56.1%) histopathologic type, which was followed by nodular sclerosing (NS, 18.3%) in frequency. None of the patients received radiotherapy as initial treatment. In 67 children the COPP regimen alone and in 15 the ABVD regimen alternating with COPP were started, to be given as a total of 12 courses. In the patients who presented with stage I-II HD the overall survival (OAS) rate and 5-year event free survival (EFS) rate were 92.3% and 77.8%, respectively. In the patients with advanced disease (stage III-IV) OAS and 5-year EFS were estimated to be 89.5% and 67.4%, respectively. No serious toxicity of chemotherapy was detected during the follow-up. In this group, clinical, epidemiological and histopathologic features of the disease showed a special pattern close to the type I pattern of HD. Regarding the survival rules and occurrence of low toxicity in our patients, results of prolonged chemotherapy alone seem to be encouraging in most of the children with HD. However, the follow-up duration is not yet sufficient to declare a clear conclusion related to the late complications.

Zinc levels of serum, plasma, erythrocytes and hair in Turkish women with anencephalic babies.

The present study revealed a significantly low zinc concentration, measured by atomic absorption spectrophotometry, in serum, plasma and erythrocytes of mothers with anencephalic babies as compared to two control groups of women. We feel that Turkey should be added to the list of geographic regions where zinc deficiency has been found together with a high rate of CNS malformations.