Haruhiko Eguchi

Enhanced cytotoxic effect of Ara-C by low intensity ultrasound to HL-60 cells.

A clonogenic assay was tested in order to determine the effects of low intensity ultrasound on HL-60 cells in the presence of cytosine arabinoside (Ara-C). HL-60 cells were exposed to ultrasound at an intensity of 0.3 W/cm2 (48 kHz). Cells were then cultured for 8 days and the number of colonies was statistically analyzed (ANOVA). Ultrasound exposure alone for 120 s resulted in no significant decrease of colonies compared to non-treated cells (P0 = 0.1426). Significant differences (P0 < 0.005) were obtained between ultrasound treated and untreated cells in the presence of various concentrations of Ara-C (2 x 10(-9), 1 x 10(-8), 2 x 10(-8), 5 x 10(-8), 1 x 10(-7) M). Morphological evaluation of ultrasound irradiated cells with scanning electron microscopy showed minor disruption of cell surface and disappearance of microvilli. These observations suggests that low intensity ultrasound altered the cell membrane thus resulting in change in Ara-C uptake into HL-60 cells.

11p15 translocations involving the NUP98 gene in childhood therapy-related acute myeloid leukemia/myelodysplastic syndrome

In a survey of childhood therapy‐related acute myeloid leukemia/myelodysplastic syndrome (t‐AML/MDS) in Japan, we found 11p15 translocations in 5 (6%) of 81 children with t‐AML/MDS. t(11;17)(p15;q21), t(11;12)(p15;q13), t(7;11)(p15;p15), inv(11)(p15q22), and add(11)(p15) were each found in one patient. Southern blotting and/or RT‐PCR analyses revealed rearrangements of the NUP98 gene in tumor samples of all five patients. Rearrangements of DDX10 were detected in t‐AML/MDS cells with inv(11), and rearrangements of HOXA9 were detected in t‐AML cells with t(7;11). The 17q21 breakpoint of t(11;17) and the 12q13 breakpoint of t(11;12)(p15;q13) coincided with the loci of the HOXB and HOXC gene families, respectively. Therefore, it is reasonable to speculate that one of the HOXB genes and one of the HOXC genes were fused to NUP98 by t(11;17) and t(11;12), respectively, in t‐AML/MDS cells. We propose that NUP98 may be a target gene for t‐AML/MDS, and that t‐AML/MDS with a fusion of NUP98 and HOX or DDX10 genes may be more frequent in children than in patients of other age groups. Genes Chromosomes Cancer 26:215–220, 1999.

Chromosome abnormalities and MLL rearrangements in acute myeloid leukemia of infants.

Of 29 infants with acute myeloid leukemia (AML), 14 (48%) had various 11q23 translocations. MLL rearrangements were examined in 21 of the 29 patients, and 11 (52%) showed the rearrangements. 11q23 translocations and/or MLL rearrangements were found in 17 (58%) of the 29 patients. While all but one of the 17 patients with 11q23/MLLrearrangements had M4 or M5 type of the FAB classification, the 12 patients without such rearrangements had various FAB types, including M2, M4, M4EO, M6 and M7. Of the 12 patients with other chromosome abnormalities or normal karyotypes, two had inv(16) or t(16;16), one had t(1;22)(p13;q13), and two had a novel translocation, t(7;12)(q32;p13). The breakpoint on 12p of the t(7;12) was assigned to intron 1 or the region just upstream of exon 1 of the TEL/ETV6 gene by fluorescence in situ hybridization. The event-free survival at 5 years for the 17 patients with 11q23/MLL rearrangements was 42.2%, and that for the 12 patients without such rearrangements was 31.3% (P = 0.5544). 11q23/MLLrearrangements have been frequently reported and a poor prognosis in infant acute lymphoblastic leukemia implied. Our study showed that while 11q23/MLL rearrangements were also common in infant AML, AML infants with such rearrangements had a clinical outcome similar to that of AML infants without such rearrangements.

Sequential Evaluation of Left Ventricular Myocardial Performance in Children After Anthracycline Therapy

This study prospectively assessed subclinical cardiotoxicity in patients undergoing chemotherapy by using the Tei index combining systolic and diastolic time intervals. A significant difference in the Tei index was observed between patients who received a low dose and those who received a moderate to high dose of anthracycline antibiotic drugs. The Tei index is a sensitive, accurate, and easy approach for detecting subclinical anthracycline cardiotoxicity.

Enhancement of cell killing of HL-60 cells by ultrasound in the presence of the photosensitizing drug Photofrin II

Enhancement of acute cell killing of HL-60 cells by low-level ultrasound in the presence of the photosensitizing drug Photofrin II was examined. HL-60 cells were exposed to ultrasound (270 kHz) at intensities of 150, 300 and 450 mW/cm2 for 60 s in the presence of Photofrin II (200 micrograms/ml). Cell survival after treatment was 49.6 +/- 5.1%, 34.5 +/- 3.1% and 27.4 +/- 3.9%, respectively. Ultrasound exposure alone at 150, 300 and 450 mW/cm2 resulted in a decrease in cell survival to 92.9 +/- 1.5%, 82.3 +/- 2.2% and 77.9 +/- 7.2%, respectively. The cell survival rate after the addition of Photofrin II alone showed no significant cell killing. These results indicate that Photofrin II was sensitized by low-level, non-thermal ultrasound to enhance the cell killing of HL-60 cells.

Sweet syndrome in a child with aplastic anemia receiving recombinant granulocyte colony-stimulating factor

Purpose: To elucidate the pathogenesis of Sweet syndrome, one patient with aplastic anemia was evaluated. Patient and Methods: A 15-year-old girl presented with intermittent fever and progressive pallor for 3 months after non-A, non-B, non-C hepatitis. Aplastic anemia was diagnosed and therapy was begun with recombinant granulocyte colony-stimulating factor (G-CSF), methylprednisolone pulse therapy, antilymphocyte globulin and cyclosporin A. There was only an increase in the neutrophil counts. We continued G-CSF therapy of 300/μg/m2 on alternate days for 7 months. At this time the white blood cell count was 10,000/μl, and the patient developed high-grade fever and a painful, erythematous, tender plaque (3×3 cm) on the left thigh. We diagnosed the lesion as a skin infection and stopped G-CSF therapy and started antibiotics. Cultures were negative. The lesion slowly resolved, G-CSF was restarted after 2 months, and 1 month later disseminated lesions occurred. Antibiotic therapy was not effective. Results: Biopsy of the lesion demonstrated infiltration of the dermis by sheets of neutrophils. We stopped G-CSF and began corticosteroid therapy. The skin lesions resolved rapidly. Conclusion: We postulated that Sweet syndrome was induced by G-CSF treatment.

Clonal and non-clonal karyotypically abnormal cells in haemophagocytic lymphohistiocytosis

We studied chromosomes in bone marrow (BM) or peripheral blood cells of nine patients with haemophagocytic lymphohistiocytosis (HLH); three of them had a family history of HLH and four others underwent concurrent Epstein‐Barr virus (EBV) infection. In addition to a large population of normal mitotic cells, karyotypically abnormal clonal cells were found in two patients, abnormal clonal cells and a nonclonal (single) abnormal cell in one, and nonclonal abnormal cells in three. All the six patients with chromosome abnormalities died of progressive disease; one of them also had EBV infection and EBV‐associated clonal proliferation. Two of three patients with EBV infection and only normal mitotic cells in BM completely recovered from the disease.

Frequent Increase of DNA Copy Number in the 2q24 Chromosomal Region and Its Association with a Poor Clinical Outcome in Hepatoblastoma: Cytogenetic and Comparative Genomic Hybridization Analysis

In a cytogenetic and comparative genomic hybridization (CGH) study of 38 hepatoblastomas, we found gain of Iq in 17 tumors (44.7%), that of 2/2q in 14 (36.8%), that of 20/20q in 9 (23.7%) and that of 8/8q in 8 (21.0%), loss of 4q in 4 (10.5%) and no DNA copy changes with normal karyotype or no mitotic cells in 11 (28.9%). Eleven tumors with 2/2q gain detected by CGH had a total chromosome 2 gain, a partial 2q gain, or a total chromosome 2 gain with an augmented partial 2q region; the common region for DNA copy gain was 2q24. Two‐color fluorescence in situ hybridization (FISH) analyses using probes covering the centromere of chromosome 2 or HOXD13 (2q31) confirmed the CGH findings, and showed that the common region for gain in 2q was centromeric to HOXD13. Event‐free survival (EFS)±standard error (SE) at 5 years was lowest in patients with 2q gain [37±15%], highest in those with no DNA copy changes [82±12%], and intermediate in those with DNA copy changes other than 2q gain [74±13%] (P=0.0549). Multivariate analysis showed that 2q gain was an independent factor predicting a poor outcome. These findings suggest the presence of a growth‐promoting gene or an oncogene in the 2q24 chromosome band, and a tumor suppressor gene in terminal 4q, which have important roles in the development and progression of hepatoblastoma.

Inversion of chromosome 11, inv(11)(p15q22), as a recurring chromosomal aberration associated with de novo and secondary myeloid malignancies: Identification of a P1 clone spanning the 11q22 breakpoint

We studied four patients with inv(11)(p15q22) associated with malignant myeloid diseases by using fluorescence in situ hybridization (FISH) with phage and cosmid probes mapped and ordered on 11q22‐24. Two of the four patients had non‐Hodgkins lymphoma or acute lymphoblastic leukemia as the primary malignancy and had received cytotoxic chemotherapy, including topoisomerase II inhibitors. The other two had de novo acute myeloid leukemia or myelodysplastic syndrome. FISH analysis showed that all 11q breakpoints were located centromeric to the MLL gene and between cosmids CN2900 and CN1323. We identified a yeast artificial chromosome (YAC) clone that spanned the inv(11) breakpoints on 11q. From this YAC, we identified a P1 clone, which included the breakpoints in at least three of the four patients. It is highly likely that the same gene on the P1 clone is rearranged in leukemic cells of each patient. This gene may be one of the targets for topoisomerase II inhibitors. Genes Chromosom. Cancer 19:150–155, 1997.

Consensus guideline for diagnosis and treatment of childhood idiopathic thrombocytopenic purpura.

A practice guideline aimed at standardizing the treatment for childhood idiopathic thrombocytopenic purpura (ITP) is presented. This consensus guideline is based on a survey carried out via a questionnaire prepared by the ITP Committee of the Japanese Society of Pediatric Hematology and sent to society members. The survey questionnaire included questions on the diagnosis of ITP submitted for the purpose of revising the ITP diagnostic guideline prepared in 1990 by the Research Group for Intractable Hematopoietic Disorders; a revised diagnostic guideline also is presented.