Toshikazu Matsui

The HOX complex neighbored by the EVX gene, as well as two other homeobox-containing genes, the GBX-class and the EN-class, are located on the same chromosomes 2 and 7 in humans

Two newly identified human homeobox‐containing genes, GBX1 and GBX2, are closely related genes, as are members of the other homeobox genes, EN‐1 and EN‐2. GBX1 and EN‐2 have been mapped to chromosome 7q36. The present study shows that GBX2 was mapped to chromosome 2q37. EN‐1 was mapped to chromosome 2q14. Moreover, two HOX complexes neighbored by the EVX gene, HOXA and HOXD, are located at chromosome 7p15‐p14 and 2q31‐q37, respectively. Thus, it is possible that these homeobox genes were linked to each other on an ancestral genome and that the ancestral chromosome segment was duplicated during evolution.

Expression of a novel human homeobox‐containing gene that maps to chromosome 7q36.1 in hematopoietic cells

A homeobox is a DNA sequence of 180 base pairs that encodes a DNA‐binding domain known as a homeodomain. The polymerase chain reaction (PCR) has been used to prepare probes of homeobox‐containing genes. We cloned and sequenced the amplified products of PCR that was performed with human genomic DNA and two primers that correspond to well‐conserved regions in homeoboxes. Fifteen kinds of homeobox gene were identified and 13 of them were assigned to HOX genes that have already been reported. Two others represented novel homeobox genes and one of them, GBX1, was mapped to chromosome 7q36.1 by fluorescence in situ hybridization. Northern hybridization of mRNA for various kinds of hematopoietic cell showed that the newly identified GBX1 gene is expressed in K562 cells and Daudi cells.

Proliferative effects of several hematopoietic growth factors on acute myelogenous leukemia cells and correlation with treatment outcome

The response of human acute myelogenous leukemia (AML) cells to four different hematopoietic growth factors (granulocyte–macrophage colony-stimulating factor (GM-CSF), interleukin-1 β (IL-1β), interleukin-3 (IL-3), and stem cell factor (SCF)) and the relationship of the proliferative response of the AML cells to treatment outcome were studied. Proliferative responses were analyzed in 79 patients with de novo AML and 19 patients with AML arising from myelodysplastic syndrome (MDS). In de novo AML, a positive proliferative response (stimulation index >2) was seen in 65 to 75% of cases. AML cells arising from MDS had a much higher incidence of proliferative response to each growth factor (79 to 90%) and a much higher level of 3H-TdR incorporation. The relationship to treatment outcome was evaluated in 79 patients with de novo AML. The patients whose leukemic cells had a positive proliferative response to any growth factor, especially IL-3 and SCF, had a poorer outcome, ie a lower complete remission (CR) rate, shorter CR duration, and shorter survival. The outcome was particularly poor in patients whose leukemic cells had proliferative responses to all four or any of the growth factors, compared to patients whose leukemic cells had no response. This increased response may be a marker of poor prognosis in patients with AML.

Alternating non‐cross‐resistant chemotherapy for non‐Hodgkin's lymphoma of intermediate‐grade and high‐grade malignancy. A pilot study

Thirty‐two patients with advanced non‐Hodgkins lymphoma (NHL) with aggressive histologic findings were treated with cyclophosphamide, doxorubicin, methotrexate with leucovorin rescue, bleomycin, vincristine, etoposide, ifosfamide, and prednisolone (CAMBO‐VIP), in which presumably non‐cross‐resistant myelosuppressive and nonmyelosuppressive agents were administered during alternate weeks for 12 weeks. To ensure the high‐dose intensity of the protocol, dose reduction and delay in treatment were minimized. Three patients were treated inadequately. Twenty‐six (89.7%) of 29 evaluable patients had a complete response, and three had a good partial response. Relapse occurred in four patients, with a median follow‐up of 29 months. The actuarial overall survival and disease‐free survival were estimated to be 87.6% and 75.9%, respectively. The CAMBO‐VIP treatment was well tolerated; myelosuppression was severe but transient and caused no serious infections. Side effects that affected dose intensity were oral ulceration, occurring in 28 patients, and blister formation under the thickened skin of palms and/or soles, followed by desquamation (5 patients). Hepatic toxicity was generally mild to moderate; it was severe in one patient. A 12‐week regimen of CAMBO‐VIP was effective for advanced NHL with aggressive histologic findings.

Interference of a methotrexate derivative with urinary oncopterin [N2-(3-aminopropyl)biopterin] measurement by high-performance liquid chromatography with fluorimetric detection

We previously reported a HPLC assay method using fluorimetric detection for the simultaneous determination of urinary N2-(3-aminopropyl)biopterin (oncopterin, a natural pteridine newly found in urine from cancer patients), biopterin and neopterin. We now have observed that an unknown substance, which may be derived from methotrexate, in urine from a patient with stomach cancer interfered with the assay of oncopterin and demonstrated that oncopterin could be completely separated from the unidentified substance by HPLC using a Nucleosil 100-5SA strong cation-exchange column. Furthermore, oncopterin was not detectable by this HPLC-fluorimetric method in urine samples from patients with stomach cancer who were not treated with methotrexate. The content of urinary oncopterin from cancer patients is supposed to be very low, with less than 1 mumol/mol creatinine. The present results indicate that the peak found with elution from the C18 column was a methotrexate-derived compound and co-eluted with the analyte oncopterin.

Interleukin-1β (IL-1β) and acute leukemia: In vitro proliferative response to IL-1β, IL-1β content of leukemic cells and treatment outcome

We evaluated the in vitro proliferative response to exogenous IL-1β in terms of tritiated thymidine (3H-TdR) incorporation in leukemic cells obtained from 119 patients with various types of acute leukemia. The content of IL-1β in leukemic cells was measured by enzyme-amplified sensitivity immunoassay. We observed a significant proliferative response to exogenous IL-1β in leukemic cells from 2766 patients with de novo AML, 129 patients with ALL, 23 patients with AUL, 812 patients with AML arising from MDS, 47 patients with myeloid crisis of CML, and 04 patients with lymphoid crisis of CML. Proliferation was marked in myeloid leukemic cells of a more premature stem cell origin. There were no significant differences in proliferative responses among the different FAB classes of de novo AML. The IL-1β content of leukemic cells was low in patients with lymphoid leukemia, but there was no significant difference among the various types of myeloid leukemia. There was no correlation between the proliferative response to exogenous IL-1β and the IL-1β content of leukemic cells. When we correlated the proliferative response to exogenous IL-1β with treatment outcome in patients with de novo AML, we found the rate of complete remission (CR) to be lower in those with a high proliferative response. We noted a longer duration of CR (p = 0.07) and of survival (p < 0.05) in patients with a low proliferative response. Thus, a high proliferative response to IL-1β in the cells of AML patients may indicate a poor prognosis.

Large-scale team-based learning for interprofessional education in medical and health sciences

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Intensive sequential post-induction therapy for adults with acute myelogenous leukemia in first remission: Long-term follow-up and results

We designed a post-induction therapy including intensive sequential therapy with non-cross-resistant drugs in an effort to prolong disease-free survival (DFS) for adults with acute myelogenous leukemia. Forty-five patients entered this study and 33 of 35 patients entering complete remission received the post-induction therapy. With a median follow-up for survivors of 3.5 years from complete remission, the actuarial 5-year DFS was 46% +/- 19% (95% confidence interval). The five-year DFS for patients over 45 years of age was comparable to that for patients under 45 years of age (50% +/- 26% vs 47% +/- 28%). Furthermore, the actuarial 5-year DFS for patients who required two courses of induction therapy was comparable to that for patients who required only one course of induction therapy (45% +/- 29% vs 50% +/- 25%). The toxicity of post-induction therapy was tolerable and no patients died during complete remission.

Lysis of autologous tumor cells by large granular lymphocytes in patients with acute leukemia in complete remission: Correlation between lytic activity and clinical outcome

In order to evaluate the effect of specific immune response on prognosis in acute leukemia, we investigated the correlation between the lysis of autologous tumor cells (ATC) by lymphocytes and prognosis. Peripheral mononuclear cells (PMC) from most patients with acute leukemia in complete remission (CR) do not exhibit cytotoxic activity against fresh-frozen ATC, although they have adequate cytotoxic activity against K562 cells. When the large granular lymphocyte (LGL) fraction was used in this study, we observed lysis of ATC in 17 (43.6%) of 39 patients with acute leukemia (12 (42.9%) of the 28 patients with acute myelogenous leukemia (AML) and 5 (45.5%) of the 11 patients with acute lymphocytic leukemia (ALL)). With regard to prognosis, the lytic activity of the LGL fraction did not reflect the duration of CR. The median CR duration in AML patients was 13 months for the lysis-positive group and 11 months for the lysis-negative group. No significant correlation was also found between lytic activity of the LGL fraction and overall survival in each patient. However, the lysis-positive group tended to have a longer survival, the median overall survival being 48 months for the lysis-positive group vs 12 months for the lysis-negative group. The prolonging of overall survival in the lysis-positive group was attributed to a high rate of induction of second remissions in this group. Long-term patient survival in the two groups did not differ.