Expression and transcriptional regulation of the PD-Ialpha/autotaxin gene in neuroblastoma.

Autotaxin (ATX) is a newly found autocrine tumor cell motility-stimulating factor. ATX is a member of the ecto-phosphodiesterase I (PD-I)/ nucleotide pyrophosphatase family. PD-Ialpha was found as a brain-type ecto-phosphodiesterase I/nucleotide pyrophosphatase. ATX and PD-Ialpha are alternative splicing products from one gene. ATX stimulates motility of A2058 melanoma cells in vitro; however, it has not been known if PD-Ialpha/ATX is expressed in naturally occurred human tumors. In this study, we examined the expression of the human PD-Ialpha/ATX gene in human neuroblastoma tumor tissues and the motility stimulating activity of recombinant ATX on neuroblastoma cells and investigated its transcriptional regulatory mechanism in a human neuroblastoma cell line. The PD-Ialpha/ATX gene was expressed in the primary tumor tissues from neuroblastoma patients to varying degrees. This gene is also expressed in the SMS-KAN neuroblastoma cell line. We identified both isoforms, PD-Ialpha and ATX, in these tumor tissues and SMS-KAN cells. The recombinant ATX stimulated the motility of SMS-KAN cells at low nanomolar concentration. We situated the promoter region, which is essential for its transcription in SMS-KAN cells, at -287 to -254 nucleotides by the promoter activity assay. The gel-shift assay revealed that there exists a nuclear protein in SMS-KAN cells that binds this region. These new insights about autocrine tumor cell motility-stimulating protein will help us to understand the metastatic mechanism of human neuroblastoma.

Targeted down-regulation of MLL-AF9 with antisense oligodeoxyribonucleotide reduces the expression of the HOXA7 and -A10 genes and induces apoptosis in a human leukemia cell line, THP-1.

The MLL gene is frequently rearranged in leukemias, and MLL chimeric proteins generated by chromosomal translocations play crucial roles in leukemogenesis. Targets of murine Mll include HOX proteins that regulate body pattern formation and hematopoiesis. However, it is not known whether or not the MLL chimeric proteins regulate the HOX gene expression in human leukemia. To address this issue, THP-1 cells, a human leukemia cell line expressing MLL-AF9, were treated with antisense oligodeoxyribonucleotide (ODN) complementary to the coding sequence of the MLL-AF9 junction. Down-regulation of the MLL-AF9 transcript was accompanied by the reduced expression of the HOXA7 and -A10 genes, but not of the HOXA2, -A4, -A5, and -A9 genes. The number of viable cells cultured with 20 microM antisense ODN for 5 days was 10-fold lower than that of the sense ODN-treated control. And the number of the annexin V-/propidium iodide- apoptotic cells in the antisense ODN-treated cells after 3 days of culture was two-fold higher than that in the control. Staining of the antisense ODN-treated cells with Hoechst 33258 showed the morphology characteristic to apoptosis. These results indicate that MLL-AF9 regulates the expression of the selected HOX genes as well as prevents the leukemic cells from apoptosis.

Expression of HOX genes, HOX cofactors, and MLL in phenotypically and functionally defined subpopulations of leukemic and normal human hematopoietic cells.

To explore the possibility that deregulated HOX gene expression might commonly occur during leukemic hematopoiesis, we compared the relative levels of expression of these and related genes in phenotypically and functionally defined subpopulations of AML blasts and normal hematopoietic cells. Initially, a semi-quantitative RT-PCR technique was used to amplify total cDNA from total leukemic blast cell populations from 20 AML patients and light density cells from four normal bone marrows. Expression of HOX genes (A9, A10, B3 and B4), MEIS1 and MLL was easily detected in the majority of AML samples with the exception of two samples from patients with AML subtype M3 (which expressed only MLL). Low levels of HOXA9 and A10 but not B3 or B4 were seen in normal marrow while MLL was easily detected. PBX1a was difficult to detect in any AML sample but was seen in three of four normal marrows. Cells from nine AML patients and five normal bone marrows were FACS-sorted into CD34+CD38-, CD34+CD38+ and CD34-subpopulations, analyzed for their functional properties in long-term culture (LTC) and colony assays, and for gene expression using RT-PCR. 93 +/- 14% of AML LTC-initiating cells, 92 +/- 14% AML colony-forming cells, and >99% of normal LTC-IC and CFC were CD34+. The relative level of expression of the four HOX genes in amplified cDNA from CD34- as compared to CD34+CD38- normal cells was reduced >10-fold. However, in AML samples this down-regulation in HOX expression in CD34- as compared to CD34+CD38- cells was not seen (P < 0.05 for comparison between AML and normal). A similar difference between normal and AML subpopulations was seen when the relative levels of expression of MEIS1, and to a lesser extent MLL, were compared in CD34+ and CD34- cells (P < 0.05). In contrast, while some evidence of down-regulation of PBX1a was found in comparing CD34- to CD34+ normal cells it was difficult to detect expression of this gene in any subpopulation from most AML samples. Thus, the down-regulation of HOX, MEIS1 and to some extent MLL which occurs with normal hematopoietic differentiation is not seen in AML cells with similar functional and phenotypic properties.

A new nitrosourea derivative for the treatment of chronic myelogenous leukemia.

A new water-soluble nitrosourea derivative, methyl 6-[3-(2-chloroethyl)-3-nitrosoureido]-6-deoxy-alpha-D-glucopyranoside (MCNU), was found to be useful for the treatment of chronic myelogenous leukemia (CML) in the chronic phase. To compare the efficacy of MCNU with that of busulfan, patients were randomized. In the 40 patients administered MCNU, the median time to the achievement of a complete remission (CR) was 50 days. This value was shorter than that observed in 37 patients administered busulfan (126 days, p less than 0.05). There were no differences in the rate of CR achieved, mortality, median time to the onset of blast crisis (BC), BC rate, or survival rate during the observation period. The overall incidence of side effects was higher for MCNU (31%) than for busulfan (15%), but the symptoms were mild, transient and tolerable for most patients. These results suggest that MCNU is a safe and valuable addition to the therapeutic repertoire for the control of CML.

Efficacy of sulbactam/cefoperazone for the treatment of infections in patients with hematologic diseases.

The efficacy and safety of sulbactam/cefoperazone (SBT/CPZ) was studied in 94 patients with severe infections and concomitant hematologic diseases. All of the study patients were included in the evaluation for safety, and 76 cases were evaluable for efficacy. Clinical efficacy was excellent in 13 cases (17.1%), good in 27 cases (35.5%), fair in seven cases (9.2%), and poor in 29 cases (38.2%). The bacteriologic eradication was 66.7% for Gram-negative bacilli and 50.0% for Gram-positive bacteria. The efficacy rate for neutropenic patients with counts less than 50 mm3 and 100 mm3 were 47.5 and 42.9%, respectively. Efficacy in patients for whom other antibiotic therapy before treatment with SBT/CPZ had been ineffective was 46.2%. Side effects were reported in one case (1.1%), and abnormal serum liver tests in five cases (5.3%); both returned to normal after discontinuation of the study medication. SBT/CPZ was an effective antibiotic for the treatment of severe infections in the presence of concurrent hematologic diseases.

The serum level of manganese superoxide-dismutase in patients with ovarian epithelial malignancy.

The serum level of a scavenging enzyme, manganese superoxide dismutase (MnSOD) was examined in 95 patients with ovarian epithelial malignancy. The MnSOD values were variable according to the differences of surgical stage, histologic type and histological grade. Factors affecting the MnSOD level were statistically analyzed. The stage and histologic type were less effective than the histological grade in determining the serum MnSOD value. The level increased significantly in accordance with the elevation of grade. Although the mechanism remains unclear, the MnSOD determination may be useful in predicting the histological grade of ovarian carcinoma.

Leiomyosarcoma of the small intestine presenting as a pelvic mass.

We present a challenging case of differential diagnosis of leiomyosarcoma of the small intestine in a patient presented with a pelvic mass. This 43-year-old Japanese woman complained of hypermenorrhea and was diagnosed as myoma uteri. She underwent partial resection of the ileum with a primary end-to-end anastomosis, omentectomy, and appendectomy, as well as a simple hysterectomy and bilateral salpingo-oophorectomy. CT and MRI indicated an intestinal tumor at the gaseous site. The histological diagnosis was leiomyosarcoma of the small intestine and leiomyoma of the uterus. Although such leiomyosarcomas are rare, they can appear as pelvic masses and must be differentiated from gynecologic disease. Preoperative CT and MRI of the abdomen were useful in obtaining the diagnosis.

Leukemia developing after 131I treatment for thyroid cancer in a patient with Werner's syndrome: molecular and cytogenetic studies.

A 40-year-old female patient with Werner's syndrome (WS) suffering from thyroid cancer and myelodysplastic syndrome (MDS) is reported. She had been diagnosed as having WS complicated with thyroid cancer seven years previously. Total thyroidectomy and radioactive iodine (131I, 100 mCi/year) therapy for seven years had slowed the progression of thyroid cancer. She suffered a sudden onset of MDS at the age of 40 years. After six months she died from overt leukemia. We found an additional chromosome aberration of chromosome 10 in the progression of leukemia from MDS.

Death education in home hospice care in Japan.

In the practice of home hospice care, death education for both patient and family is extremely important, although little information on its usefulness is available. In this study, the effects of death education under home hospice care were analyzed for 16 patients who died at home. Death education for the patient and his/her family was given at least once in each phase of care, and at least four times in total. The acceptance of death by the patients was judged according to the way they spent their remaining time, to their attitudes, and to their hope for a life after death. Fourteen of 15 patients appeared to accept their own death. An autopsy was performed in five of the 16 cases. In one case, the doctor recommended an autopsy to the family; in the other cases, it was performed in accordance with the patient's or family's wish. As the goal of death education in home hospice care is the acceptance of death by both patient and family, our methods of death education appear to be effective.

[Actual conditions of bacterial infection associated with hematopoietic disorders--changes in 10 years. The Hanshin Study Group of Hematopoietic Disorders and Infections].

This study showed several accumulated data through ten years from our experience in hematopoietic disorders and associated infections, which has been analyzed by the Hanshin Study Group of Hematopoietic Disorders and Infections. Since 1979 to 1988, our group had evaluated the sorts of causative organisms and the efficacy of various antibiotics therapy in 2119 cases of infectious diseases associated with hematopoietic disorders. On behalf of evaluating the changes of disease profile for ten years, we divided the accumulated data into three phases; former phase the first three years, middle phase the second three years and late phase the last four years. There was no significant difference in the frequency of various hematopoietic disorders among the three phases. Each leukemia patients occupied 77% of all cases. Sepsis suspected is the most frequent infectious disease accounting for 68.8%. The other infectious diseases were 8.4% of the sepsis, 14.8% of the respiratory infections and 3.1% of the urinary tract infections. Comparing the frequency of infections among the three phases, the respiratory and urinary tract infections inclined to decrease. Of the 532 strains isolated from 2119 cases and identified as causative organisms, gram-negative bacilli occupied 62.8% and gram-positive bacteria 36.5%. In comparing the percentage of gram-negative bacilli among the three phases, it showed a decreasing tendency in order former phase 63.6%, middle phase 76.4% and late phase 43.8%. Pseudomonas, however, had been isolated at almost constant ratio through ten years. On the other hand, the ratio of gram-positive bacteria isolated were 34.5% in former phase, 23.6% in middle phase and 56.3% in late phase, showing increasing a tendency through the period. Twenty-three kinds of antibiotics were administered by intravenous drip infusion. The efficacy rate was 43.9% to 67.2%. In particular, effectiveness of antibiotic therapy often depends on the change of peripheral neutrophil counts from the onset and during the therapy. The efficacy rate, however, was 36% even neutrophil counts have not shown the tendency of increase from less than 100/microliters.

[Efficacy and safety of cefbuperazone in severe infections complicating hematologic diseases Hanshin Infection Study Group].

Cefbuperazone (CBPZ) was administered to patients with severe infections complicating hematologic diseases to assess its efficacy and safety under such clinical conditions. Primary diseases in this series of 78 cases included; acute leukemia in 41 cases, chronic leukemia in 6 cases, other leukemia in 9 cases, malignant lymphoma in 13 cases, multiple myeloma in 3 cases, aplastic anemia in 5 cases and 1 other case. Types of infection included sepsis; proven or suspected, in 59 cases, pulmonary infection in 8 cases, upper respiratory infection in 5 cases, and other cases. CBPZ was infused by an intravenous drip method at a dosage of 4-8 g daily. Patients' ages ranged from 14 to 85 years. Clinical response to the CBPZ regimen was excellent in 24 cases, good in 22 cases, fair in 2 cases, and poor in 30 cases. Thus the overall efficacy rate (percentage of cases showing an excellent or good response) was 59.0%. Efficacy rates for individual types of infection were: documented sepsis 16.7%, suspected sepsis 58.5%, lower respiratory infection 62.5%, and upper respiratory infection 100%. CBPZ also proved to be effective in 61.0% of cases with a neutrophil count of less than 500/mm3 prior to therapy. Side effects encountered were diarrhea in 1 case, gastric discomfort in 1 case and hepatic dysfunction in 5 cases. These side effects, however, were not dose-related, and none were serious. These results indicate that CBPZ has a high therapeutic efficacy even in patient with compromised immunodefenses.

[Therapeutic effects of cefuzonam against severe infections in patients with hematopoietic disorders. Hanshin Infection Study Group].

Cefuzonam (CZON) was used to treat severe infections in 151 patients with hematopoietic disorders, and its efficacy and safety were assessed. The drug was given in doses of 2.0 to 6.0 g a day, divided into 2 or 3, intravenously by injection or infusion. The clinical effects were excellent in 34 cases, good in 40 cases, fair in 5 cases, and poor in 57 cases. Therefore, the results were excellent or good in 54.4% of the patients treated. The efficacy rates were 43.8 and 35.9% for groups of patients whose neutrophil counts were 500/microliters or less and 100/microliters or less, respectively. It was excellent or good in 70.6% of patients in whom causative agents were identified, and in 66.7 and 80.0% of patients infected with Gram-negative and -positive bacilli, respectively. The efficacy rate for patients infected with unidentified agents was 52.1%. The rate for patients who had received other antibiotics previously was 41.5%. The rate for patients having received only one antibiotic for the preceding treatment was 50.0%. Six (3.9%) of the treated patients experienced adverse effects including changes in laboratory test results observed in 4.

[Therapeutic effects of meropenem against severe infections in patients with hematopoietic disorders. Hanshin Study Group of Hematopoietic Disorders and Infection].

The efficacy and the safety of meropenem (MEPM), a newly developed carbapenem antibiotic, were investigated in 150 patients with severe infections complicated with hematopoietic disorders. 1. Clinical responses in 132 patients who were evaluable for effectiveness were excellent in 33 patients, good in 45, fair in 10 and poor in 44, with an efficacy rate of 59.1%. 2. The efficacy rate in patients who had previously been treated with the other antibiotics was 51.2%, while that in patients who had not been thus treated was 62.9%. 3. The efficacy rate in patients whose neutrophil counts increased during the therapy with MEPM was higher than that in patients whose neutrophil counts did not increase. The efficacy rate in patients whose neutrophil counts during the therapy were below 100/mm3 was 48.1%. 4. Out of 150 cases, side effects were observed in 4 patients, 3 with eruption and 1 with jaundice. Abnormalities in laboratory test results on liver functions were noted in 8 patients. These results indicate that MEPM is an effective and safe antibiotic for the treatment of severe infections in patients with hematopoietic disorders.

[Therapeutic effect of cefotaxime against severe infections in patients with hematopoietic diseases].

Seventy-five patients with severe infection accompanying hematologic disorder, including leukemia and malignant lymphoma, were treated with cefotaxime (CTX). CTX was administered by intravenous drip infusion at a daily dose ranging from 4 to 16 g for terms of 3 to 21 days. The total doses were ranged from 12 to 226 g. The results obtained were as follows: Clinical effects: Excellent in 20 cases, good in 21 cases, fair in 7 cases and poor in 27 cases. The efficacy rate was 54.7% (41/75). Clinical effectiveness on isolated organisms (27 cases): In single infection (21 cases), the efficacy rates were 80% for Gram-positive cocci, including S. aureus and 63.6% for Gram-negative bacilli other than P. aeruginosa. In mixed infection (6 cases), the rate was 50.0%. There were no significant differences in the efficacy rates for those patients who were grouped by the initial number of neutrophil (less than 100, 101--500 and over 501/mm3). There were no significant difference in the efficacy rates for those patients who were grouped by the initial number of lymphocyte (less than 500 and over 501/mm3). Side effects and abnormal laboratory findings: One case of skin rash and 2 cases of elevated GOT and GPT were observed. CTX was therefore considered as a clinically useful antibiotic for the severe infections even in neutropenic state in patients suffering from malignant hematological diseases.

[Therapeutic effectiveness of ceftizoxime on severe infections associated with hematologic disorders].

One hundred patients with severe infections associated with hematologic disorders, including leukemia and lymphoma, were treated with ceftizoxime (CZX) in daily doses of 4 approximately 9 g for an average of 8.9 days. In the 84 patients who completed the trial, response was excellent in 27 (32.1%) and moderate in 25 (29.8%). The rate of effectiveness was 61.9%. The only side effect seen during the treatment was skin rash in 3 patients. Hepatic disorders were observed in 5 patients. The relation between CZX and these abnormal findings was not established. These results indicate that CZX is a therapeutically effective and safe antibiotic for the treatment of severe infections in patients with underlying hematologic disorders.

[Therapeutic effectiveness of aztreonam on severe infections associated with hematologic disorders. Hanshin Infection Study Group].

One hundred and nine patients with infection accompanying hematologic disorders including leukemia and lymphoma were treated with aztreonam (AZT). Of the 90 patients in whom the efficacy could be evaluated, 17 (18.9%) responded markedly and 29 (32.2%) moderately, the effective rate being 51.1%. The efficacy rate classified according to infections was 25% in septicemia, 46.3% in suspected septicemia, 57.1% in pneumonia and 100% in urinary tract infection. The efficacy rate to the Gram-negative bacteria was 78.9% and to the Gram-positive bacteria was 20.0%. In 4 (66.7%) out of 6 patients in whom P. aeruginosa was the causative organism, AZT was effective. The efficacy rate was 52.2% in the 23 patients whose causative organisms were identified and 50.7% in 67 patients whose causative organisms were not identified. There was no significant difference in the efficacy rate between the patients who failed to respond to prior antibiotic therapy (53.6%) and those treated with AZT from the beginning. The initial neutrophil count did not affect the efficacy rate. Side effects which might have been caused by AZT were eruption and fever in 4 patients. Hepatic disorders and eosinophilia were observed in 7 patients. However, the relationship between AZT and these abnormal findings was not established. These results indicate that AZT is an effective and safe antibiotic for the treatment of infections accompanying hematologic disorders.

[Effects of cefoxitin in the treatment of severe infections in patients with hematopoietic disorders].

Cefoxitin (CFX) at a daily dose of 3 to 12 grams was administered to patients who had hematopoietic disorders as underlying diseases and having severe infections. Efficacy and safety of the drug were evaluated. The underlying diseases in the 64 patients included in the evaluation of efficacy were acute myelocytic leukemia (30 cases), acute lymphocytic leukemia (9), acute promyelocytic leukemia (3), acute monocytic leukemia (2), chronic myelocytic leukemia-blastic crisis (10), erythroleukemia (2), malignant lymphoma (2), aplastic anemia (2), and others (4). The infections were septicemia in 3 patients, suspected septicemia in 47, respiratory tract infections in 7, oral infections in 3, urinary tract infections in 2, and others in 2. The clinical efficacy of CFX was 'excellent' in 13 patients, 'good' in 26, 'fair' in 6, 'poor' in 19 for an efficacy rate of 60.9%. The efficacy rate classified according to infections was 66.7% in septicemia, 66.0% in suspected septicemia, 42.9% in respiratory tract infections and 66.7% in oral infection. The organisms isolated from the patients with septicemia were E. coli in 2 patients and B. cereus in 1. B. cereus was not susceptible to CFX. The efficacy rate was 60.0% in the 10 patients whose causative organisms were identified and 61.1% in the 54 patients whose causative organisms were not identified. There was no significant difference in the efficacy rate between the patients who had failed to respond to prior antibiotic therapy and those treated with CFX from the beginning. The efficacy rates for the former group (23 patients) and for the latter group (41 patients) were 56.5% and 63.4%, respectively. The efficacy rate in patients with an initial neutrophil count less than 500/mm3 (35 cases) and from 501 to 1,000/mm3 (13 cases) were 57.1% and 76.9%, respectively. Side effects which might have been caused by CFX were skin eruptions in 2 patients (2.6%) and transient elevation of GOT and GPT in 1 patient (1.3%) among 76 patients who were evaluated for safety. CFX was considered to be a markedly useful and safe drug in the treatment of patients with hematopoietic disorders who developed severe infections.

[Cooperative studies on clinical effect of cefotetan for severe infections combined with hematologic disorders].

Seventy-six patients with severe infection accompanying hematologic disorders including leukemia and lymphoma were treated with intravenous drip infusion of cefotetan (CTT). Of the 66 cases in whom the efficacy could be evaluated, 22 cases responded markedly and 16 cases moderately, the effective rate being 57.6%. It is impressed that more cases responded markedly to CTT than to any other antibiotics previously studied. None of the cases revealed serious side effects attributable to CTT. These results indicate that CTT is an effective and safe antibiotic for the treatment of severe infection accompanying hematologic disorders. As CTT was administered to special cases under marked decrease of neutrocyte, the importance of neutrocyte in the treatment of infection was also indicated.

[Therapeutic effect of ticarcillin against severe infection in patients with hematologic malignancy (author's transl)].

Seventy patients with severe infection accompanying hematologic disorders including leukemia and lymphoma were treated with ticarcillin in combination with aminoglycosides and/or cephem derivatives. Of the 58 patients in whom the efficacy could be evaluated, 17 (29%) responded markedly and 22 (38%) moderately, the effective rate being 67%. Side effects attributable to the treatment were noted: rash in 2, fever and rash in 1, vascular pain in 2 (5 and 7 years of age). Renal and hepatic functions were abnormal in 4 each. However, these abnormal findings were not attributed to ticarcillin. These results indicate that ticarcillin is an effective and safe antibiotic for the treatment of severe infection accompanying hematologic malignancy when used in combination with aminoglycosides and/or cephem derivatives.

[Clinical evaluation of high dose intravenous injection of fosfomycin on the severe infections associated with the treatment of haematological disorders].

Fosfomycin (FOM) was administered intravenously to 65 cases of severe infections complicated with 62 cases of several haematological disorders. Out of 65 cases, 45 were treated with high doses FOM, i.e., 8 g per day or more. Another 20 cases were treated in usual doses of 4--6 g per day. Causative organisms were isolated from 52 cases of which 32 cases were Gram-negative bacilli and 19 cases were Gram-positive cocci. The effective rate of FOM was 57.8% in the high-dose treatment group (26/45) and 45.0% in the usual-dose treatment group (9/20), but the significant difference was not defined. Among 32 cases with Gram-negative bacilli infections, including Pseudomonas aeruginosa or Serratia marcescens, 15 cases were effective (47%). On the contrary, 14 out of 19 cases with Gram-positive cocci infections were effective (74%). Thirteen cases (50.0%) were effective even in which neutrophils were less than 500/cmm before FOM administration. Severe side effects were not observed, without 2 cases. One was skin eruption due to drug allergy and the other was suspected to be interstitial pneumonitis, but not confirmed pathologically. These data suggest that high dose treatment of FOM were useful for the severe infections even in neutropenic state in haematological disorders.