[L-Asparginase effect with 6000U/m2 on lipid profile in children with acute lymphoblastic leukemia]

One of the major etiologies of lipid disorders in malignancies is the use of chemotherapy drugs, the most important of which is L-Asparginase. Studies in different centers with high dosages of L-Asparginase demonstrated different results. The aim of this study is to evaluate the effect of L-Asparginase at a dose of 6000 U/m2 on lipid profile in Iranian children with newly diagnosed acute lymphoblastic leukemia in Mofid Children Hospital of Tehran. We performed a nonrandomized trial in which all children with newly diagnosed acute lymphoblastic leukemia [ALL] aged <15 years participated. Every case serves as his/her own control. The cholesterol, triglyceride [TG], HDL, LDL, VLDL, APOA, APOB and LPa were evaluated in three stages: before, during and 2 months after treatment with L-Asparginase after which the results were compared. In our study, 82 newly ALL diagnosed patients with the mean age of 6 years [within the age range of 0.12-14 and SD of 3.5] were evaluated. Mean seum level of TG in pretreatment stage was 163.9 mg% and during treatment with L-Asparginase 123 mg% demonstrating significant decline in TG serum level after administration of L-Asparginase [p=0.002]. Mean serum level of cholesterol before administration of L-Asparginase was 151mg% and during treatment 140 showing no statistically significant difference [p= 0.061]. LPa level in pretreatment phase was 24mg% and during treatment 14 showing a statistically significant difference [p= 0.0001]. L-Aspar resulted in decline in TG serum level and increase in HDL though with no significant difference in cholesterol level. Overall, L-Asparginase even at a dose of 6000U/M2 does not raise blood level of triglyceride and cholesterol

Vincristine induced neurotoxicity: Study of 75 cases

Concern for side-effects of therapy related to treatment of childhood malignancies is becoming an increasingly important topic. In this study, we evaluated extent of vincristine [VCR] induced neurotoxicity in a group of children who underwent chemotherapy, with VCR being part of the regimen. In this investigation, for 75 children [54% boys, 46% girls], aged between 1 to 14 [mean 6.5 +/- 4.3] years, serial weekly neurological examinations were performed; of the 75, 70 had acute lymphoblastic leukemia and 5 Wilm's tumor. All patients were on a chemotherapy protocol of at least 4 consecutive VCR [1.5 mg/m[2]] injections. Decreased deep tendons reflexes were seen in the Achilles reflex in 78%, and the patellar reflex in 53% of patients. Muscle weakness was found in 70% of patients, being mild in 76% of them. Four percent of patients showed severe weakness. Petosis, jaw pain, hoarseness, abdominal pain and constipation were seen in 15%, 6%, 12%, 12% and 12% respectively. Paresthesia was observed in 32 of 52 patients, over 4 years old. No cases of foot drop, urinary retention or facial nerve palsy were seen in this patient group. Children on usual doses of vincristine regimen may have neuropathic side effects but most of these side effects are mild and not troublesome

Griscelli Syndrome; A case report and review of the literature

Griscelli syndrome [GS] is a rare disease first described in 1978. It is inherited in autosomal recessive pattern. This disease is characterized by partial albinism, pigmentation dilution, cellular immunodeficiency, neurological involvement and uncontrolled phases of macrophage and lymphocyte activation. We report a 5 months Old Iranian girl presenting with silver-gray hair, eyelashes and eyebrows, hepatosplenomegaly, pancytopenia, hemophagocytosis and progressive neurologic deterioration. Griscelli syndrome can be suggested according to her symptoms. The chemotherapy was not effective for her and she died due to multi organ failure

Use of immunoglobulin heavy chain and kappa light chain gene rearrangements by PCR for molecular diagnosis of minimal residual disease in Iranian children suffering from beta-precursor acute lymphoblastic leukemia

Diversity of IgH and IgK molecules is generated during B and T Lymphocyte differentiation through the rearrangement of variable, diversity, junction and constant gene segments. Additionally, random insertion and deletions of nucleotides between gene segments make unique sequences which are cell or clone specific. Similar IgH and IgK genes rearranged in normal cells of lymphoid leukemia cases can be used as a marker of clonality and for evaluation of minimal residual disease [MRD]. The purpose of this study is to evaluate the pattern of IgH chain and IgK gene rearrangements using polymerase chain reaction [PCR] in beta-precursor acute lymphoblastic leukemias [ALL] to follow the MRD at day 14, day 28 [end of remission induction], week 10, 3-6 months and 6-12. month after the initiation of treatment. In our prospective study bone marrow aspirates of 183 children at the mean age of 63.6 months with diagnosis of acute leukemia were collected at admission before any chemotherapy. After reviewing cytomorphology and immunophenotyping, only 140 cases with diagnosis of beta-precursor ALLs were selected for study. Mononuclear cells including leukemic blasts were isolated by density gradient. After DNA extraction, IgH and IgK [V[K] I-IV / Kde] were amplified by consensus primers using PCR. PCR products were analyzed after heteroduplex analysis and polyacrylamide gel electrophoresis [silver stain]. The DNA sequences were compared and aligned with the sequences homologous for IgH and IgK published by Gene Bank. The follow up specimens were collected at day 14, day 28 [end of remission induction], day 45-month 3, and 3-6 months and 6-12 months after initiation of treatment. After routine cytomorphologic analysis, similar PCR was done on follow up extracted DNAs in parallel with diagnosis DNA. MRD was considered to be approved positive if bands similar to those at the time of diagnosis were present. Statistical analysis using SPSS software [version 11.5] was performed. 90.5% of patients had clonal IgH gene rearrangements. Monoclonal, biclonal and oligoclonal patterns were observed in 57.8%, 34.9% and 5.5% of patients with IgH [CDR III] rearrangement, respectively. Clonal patterns of IgK-Kde were detected in 59 [67%; n: 88] of BP-ALLs. According to cytomorphology about 92% of patients were in complete remission. MRD positivity decreased from more than 90% to 20% using different gene rearrangements in defined time points. Four patients who relapsed during follow up were MRD positive using 1-3 rearrangements and all except one were in clinical remission. Clonal rearrangement of IgH had a pattern similar to other populations. IgK was slightly more frequent than previously reported and the VKI [25%] was the most common type. These differences can be explained by different techniques, DNAs and clonality markers. According to the results, these clonal markers can be used in diagnosis and follow up of MRD

[Prevalence of Helicobacter pylori infection in children with chronic immune [idiopathic] thrombocytopenic purpura and evaluation of Helicobacter pylori eradication on platelet count]

Recently, the high prevalence of Helicobacter pylori infection has been reported in adult patients with chronic immune [idiopathic] thrombocytopenic purpura. Furthermore, after Helicobacter pylori eradication therapy in such patients, their platelet counts have been observed to increase, suggesting that Helicobacter pylori may be a causative agent of adults' chronic idiopathic thrombocytopenic purpura. However, there have been only a few reports of this subject in children with chronic thrombocytopenic purpura. The purpose of this study is to determine prevalence of Helicobacter pylori infection in Iranian children with chronic thrombocytopenic purpura and role of Helicobacter pylori eradication in rising platelet count of these patients. This descriptive-clinical trial study was performed in 31 children under 14 years old with chronic thrombocytopenic purpura who attended hematology ward of Mofid paediatric hospital. After determining platelet count, and filling the results patients referred to gastrointestinal ward of the hospital to perform urea breath test for evaluation of Helicobacter pylori infection, then Helicobacter pylori-infected patients who were diagnosed by this test, received eradication therapy using triple therapy regimen [containing Omeperasole, Amoxicillin and Clarithromycin] for 2 weeks and their platelet counts were recorded during the follow up period. Mean age of the patients was 8.9 +/- 3.2 years old ranging from 3.5 to 14 years old. They were 17 [54.9%] girls and 14 [45.1%] boys. Mean platelet count of the patients was 51.4 +/- 34.3x10[9] / L ranging form 125x10[9]/ L to 8x10[9] / L. Mean duration of disease in the patients was 27.7 +/- 20.2 months ranging from 7 to 96 months. Helicobacter pylori infection was found in only 4 children [12.9%] and Helicobacter pylori eradication therapy was not effective in rising platelet count to achieve complete or partial remission. Comparing Helicobacter pylori-positive and negative patients, there were no significant differences regarding their age, platelet count and duration of disease. This study shows that prevalence of Helicobacter pylori infection in children with chronic immune thrombocytopenic purpura is less than that is in adults. Furthermore, we have found that platelet count in Helicobacter pylori-positive children have not been risen after eradication therapy. We suggest that more studies in different gender groups and different zones in the world with more number of samples should be performed, especially in children in order to determine both the exact role of Helicobacter pylori's pathogenesis in developing the chronic idiopathic thrombocytopenic purpura and the effectiveness of eradication therapy in rising platelet count in these patients

[Molecular diagnosis of clonality in B-precursor acute lymphoblastic leukemia of children using immunoglobulin heavy and kappa light chain gene rearrangements]

Enormous diversities of heavy chain immunoglobulin [IgH] and IgK molecules are generated during B- and T-lymphocyte differentiation through the rearrangement of gene segments. Additionally, random insertion and deletion of nucleotides between gene segments make unique sequences which are cell or clone specific. IgH and IgK gene rearrangements are the most and relatively common reported rearrangements in B-precursor acute lymphoblastic leukemia, respectively. The purpose of this study is to evaluate the pattern of IgH and IgK gene rearrangements using polymerase chain reaction [PCR] in BP-ALL in Iranian children. For this prospective study, bone marrow aspirates of 183 patients with the diagnosis of acute leukemia were collected at admission before any chemotherapy. Having reviewed cytomorphology [L[1]:44%, L[2]:41%] and immunophenotyping, only 140 cases with the diagnosis of B-precursor ALL were selected. Mononuclear cells including leukemic blasts were isolated by density gradient. Having DNA extracted, hyper-variable regions of IgH and IgK were amplified by consensus primers using PCR. PCR products were analyzed after heteroduplex analysis and polyacrylamide gel electrophoresis [silver stain]. The DNA sequences were compared and aligned to the sequences homologous for IgH and IgK published by Gene Bank. IgH gene rearrangements were found in 114 [90.4%] of patients using consensus primers for CDR-III and CDR-I regions [monoclonal 57.8% biclonal 34.9% oligoclonal 5.5%]. Four of nine patients with T-ALL had clonal rearrangements of IgH. Clonal pattern of Ig?-Kde were present in 59 [67%] of cases [biclonal 10%]. VKI [25%] and VK? [22.7%] were the most common type of rearrangements. Clonal rearrangement pattern of IgH gene was similar to other populations. Using FRI and FRIII primers in multiplex PCR increased the rate of detection and reducing turnaround time. IgK was slightly more frequent than previous reports while VKI [25%] was the most common type

pattern of cross lineage T-cell receptor delta/gamma gene rearrangements in B-precursor acute lympho-blastic leukemia of Iranian children using polymerase chain reaction

Diversity in heavy chain immunoglobulin [IgH] and T-cell receptor [TCR] molecules occures during B- and T-lymphocyte differentiation through the rearrangement of variable [V], diversity [D], junction [J] and constant [C] gene segments. Lymphoid leukemia cells are similar to normal precursors and have rearranged IgH, IgK and TCR [cross-lineage rearrangement] genes which can be used as a marker of clonality and evaluation of minimal residual disease [MRD]. The purpose of this study is to evaluate the pattern of TCR- delta/gamma gene rearrangements using Polymerase Chain Reaction [PCR] in B-precursor acute lymphoblastic leukemia [ALL] in Iranian children. In our prospective study, bone marrow aspirates of 183 children with early diagnosis of acute leukemia were collected at admission before any chemotherapy. After reviewing cytomorphology and immunophenotyping, only 140 subjects with diagnosis of B-precursor ALLs were selected for study. Sixteen were excluded from our study due to various reasons including cellular degeneration. Mononuclear cells including leukemic blasts were isolated by density gradient. After DNA extraction, hyper-variable regions TCR-delta [V delta2-D delta3 and D delta2-D delta3] and TCR-gamma [V gamma; V gamma I and V gamma II] were amplified by consensus primers using PCR. PCR products were analyzed after heteroduplex analysis and polyacrylamide gel electrophoresis [silver stain]. The DNA sequences were then compared and aligned to the homologous sequences of Gene Bank for confirmation. T-test, Mann whitney, Fisher exact test and Chi-square were used for data analysis. Clonal rearrangement of TCR-gamma [V gamma] and V gamma l/Il were present in 79.3% and 64.9% of patients respectively and only 5% of cases showed biclonal pattern. The V gamma ll rearrangement was the most common [46.8%] type in TCR-gamma. 47 [45.2%] and 11 [16.6%] of patients had V delta2- D delta3 and D delta2-D delta3 partial gene rearrangements, respectively. Biclonal/oligoclonal patterns were present respectively in 27.7% and 4.3% of cases with Vdelta2-D delta3 rearrangement. Only one patient had biclonal D delta2 D delta3 rearrangement. Clonal rearrangement of TCR-delta [Vdelta2-Ddelta3 and D delta2-D delta3] genes had a pattern similar to other populations. Frequency of TCR- gamma [V gamma I and V gamma II] rearrangements was slightly higher than previous reports, and in contrary to others except for Brazilian report the V gamma II rearrangement was the most common type. We found no significant correlation between presence of different types of rearrangements and quantitative variables. The only significant point was the reduction of Vdelta2Ddelta3 with increase in age. According to preliminary results, these clonal markers can be used in diagnosis and follow up of MRD

Recurrent bilateral spontaneous pneumothorax in early infancy: a case of langerhans cell histiocytosis

Langerhans cell histiocytosis [LCH] is a rare disorder characterized by infiltration of either single or multiple organs by S100 and CD1a positive cells. Patients with pulmonary LCH are predisposed to pneumothorax due to destructive changes in the lung parenchyma. Here, we report a case of multisystem LCH who presented at 2 months of age with simultaneous bilateral spontaneous pneumothorax

Evaluation of bi/oligoclonal rearrangement of immunoglobulin and T-cell receptor genes in Iranian children suffering B-precursor acute lymphoblastic leukemia

Clonal gene rearrangement of immunoglobulin and T cell receptor may have mono, bi or oligoclonal pattern. Significance of these patterns were studied at diagnosis and follow up of MRD in many countries, however, similar studies have not been conducted among Iranian patients. We investigated the bi/oligoclonal pattern and their association with quantitative and qualitative parameters especially MRD in Iranian children suffering from B-precursor acute lymphoblastic leukemia. In our prospective study, bone marrow aspirates of 140 patients with B-precursor ALLs were selected. Mononuclear cells including leukemic blasts isolated by density gradient. Having DNA extracted, hypervariable regions of IgH, IgK, TCR-delta [D delta 2-D delta 3, V delta 2-D delta 3] and TCR-lambda [V lambda, V lambda I, V lambda II] were amplified by consensus primers using PCR. PCR products were analyzed after heteroduplex analysis and polyacrylamide gel electrophoresis [silver stain]. The DNA sequences were compared and aligned to the sequences homologous for IgH and IgK published by Gene Bank. Bone marrow aspirates of days 14, 28 and 45, as well as months 3 and 6 were treated similarly. IgH gene rearrangements were reported in 114 [90.5%] patients using consensus primers for CDR-III and CDR-I regions [monoclonal: 57.8%, biclonal:34.9% and oligoclonal:5.5%]. Clonal pattern of IgK-Kde were present in 59 cases [67%] [biclonal:10%] Clonal rearrangement of TCR-lambda [V lambda] and V lambda I/II were present in 79.3% and 64.9% of patients, respectively, however, only 5% of cases showed biclonal pattern. The V lambda II rearrangement was the most common [46.8%] type in TCR-lambda. 47 [45.2%] and 11 [16.6%] patients had V delta 2-D delta 3 and D delta 2-D delta 3 partial gene rearrangements, respectively. Biclonal/oligoclonal pattern were present in 13 [27.7%] and 2 [4.3%] cases with V delta 2-D delta 3 rearrangement. Only one patient had biclonal D delta 2-D delta 3 rearrangement. No significant difference regarding the quantitative and qualitative parameters and MRD was observed between the two groups. Bi/oligoclonal rearrangement of IgH, IgK, TCR-delta [D delta 2-D delta 3, V delta 2-Ddelta 3] and TCR-lambda [V lambda, V lambda I, Vlambda II] genes had comparable pattern to other populations. Results of MRD study showed no significant differences between the two groups

[Prevalence of prognostic factors in children with Acute Lymphoblastic Leukemia admitted to Mofid Children's Hospital from 1997-2000]

Acute leukemia is the most common malignancy in children and acute lymphoblastic leukemia [ALL] accounts for 75% of acute leukemia cases. New treatment protocols have resulted in complete remission rates up to nearly 100% in children with acute lymphoblastic leukemia. Today, one of the most important prognostic factors in acute lymphoblastic leukemia is intensity of the treatment. Risk stratification is accomplished based on clinical, morphological, immunophenotypic and cytogenetic findings. The aim of this study was to determine some prognostic factors in children with acute lymphoblastic leukemia. In this retrospective study information about age at onset of acute leukemia, sex, initial white blood cell count, FAB-subtype, immunophenotype, and clinical course of newly diagnosed acute lymphoblastic leukemia were extracted from medical records of children admitted to pediatric oncology department of Mofid Children's Hospital from 1997 to 2000. There were 81 [58.3%] male and 58 [41.7%] female. 3.6% of patients were under 1 year old, 18.7% were above 10 years and 77.7% were 1-10 years old. Initial WBCs more than 50,000/ mm3 were observed in 24.5% of patients. 91.4% of patients showed FAB-subtype of L1, 7.9% of L2 and L3 was detected just in 0.7% of cases. Lymphoblasts were of B-cell lineage in 92.7% of patients, with pre-B cell in 74.8%, early pre-B cell in 17.1% and mature B cell in 0.8%. As a whole, complete remission was observed in 79.3% of the patients. 12.9% of patients had a relapse, second remission was achieved in 2.6% of relapsed cases. In this study, FAB-subtype L1 and pre-B cell immunophenotype was more common than previous studies. Other results were the same as reported in older studies