The anemia of the newborn induces erythropoietin expression in the developing mouse retina.

The hematopoietic hormone erythropoietin (Epo), regularly produced by the kidneys and the liver, is also expressed in neuronal tissue, where it has been found to mediate paracrine neuroprotective effects. In most studies exploring the rescue effects of Epo, apoptosis was exogenously induced by different cell death stimuli. Herein, we set out to study the expression and function of Epo in physiologically occurring apoptosis in a model of retinal development. We made use of an organotypic retinal wholemount culture system that resembles the physiological in vivo situation with cell connections still retained. Epo mRNA expression in the retina, liver, and kidney showed a significant increase during early development, coinciding with the anemia of the newborn. In the retina of Epo-green fluorescent protein transgenic mice, Epo-expressing cells were identified and found to be distributed in the retinal ganglion cell layer. Treatment of retinal wholemount cultures with recombinant Epo resulted in a significant decrease of apoptotic ganglion cells as well as photoreceptor cells throughout retinal development. Moreover, transforming growth factor-beta-induced apoptosis was completely antagonized by Epo when both factors were simultaneously applied. Investigations on the signaling pathway revealed a decrease in Bax mRNA levels in Epo-treated retinal cells. We conclude that Epo exerts wide and prolonged neuroprotective activity in physiologically occurring apoptosis and thus contributes to proper retinal development.

Caffeine and taurine enhance endurance performance.

Caffeine enhances endurance performance; however, its effect on accumulated lactate remains unclear. Conversely, taurine, which also enhances endurance performance, decreases accumulated lactate. In this study, the effect of combination of caffeine and taurine on endurance performance was assessed. Mice ran on a treadmill, and the accumulated lactate was measured. In addition, muscle fibers from the gastrocnemius muscle of the mice were stained with ATPase and analyzed. The use of caffeine and taurine over a 2 week period enhanced endurance performance. Moreover, taurine significantly decreased the accumulated concentration of lactate over long running distances. However, the diameter of the cross-sections and ratios of Types I, IIA, and IIB muscle fibers were not affected.

Characterization of stage progression in chronic myeloid leukemia by DNA microarray with purified hematopoietic stem cells.

Chronic myeloid leukemia (CML) is characterized by the clonal expansion of hematopoietic stem cells (HSCs). Without effective treatment, individuals in the indolent, chronic phase (CP) of CML undergo blast crisis (BC), the prognosis for which is poor. It is therefore important to clarify the mechanism underlying stage progression in CML. DNA microarray is a versatile tool for such a purpose. However, simple comparison of bone marrow mononuclear cells from individuals at different disease stages is likely to result in the identification of pseudo-positive genes whose change in expression only reflects the different proportions of leukemic blasts in bone marrow. We have therefore compared with DNA microarray the expression profiles of 3456 genes in the purified HSC-like fractions that had been isolated from 13 CML patients and healthy volunteers. Interestingly, expression of the gene for PIASy, a potential inhibitor of STAT (signal transducer and activator of transcription) proteins, was down-regulated in association with stage progression in CML. Furthermore, forced expression of PIASy has induced apoptosis in a CML cell line. These data suggest that microarray analysis with background-matched samples is an efficient approach to identify molecular events underlying the stage progression in CML.

Morphological changes in human gastric tumours after eradication therapy of Helicobacter pylori in a short-term follow-up.

BACKGROUND: It is controversial as to whether the development of gastric cancer is influenced by Helicobacter pylori eradication. If eradication itself influences the tumour morphology, this may affect the tumour discovery rate. AIM: To investigate the morphological changes in the gastric neoplasm after H. pylori eradication. METHODS: We studied 37 patients with eradication therapy. After a 1-month follow-up, endoscopic re-evaluation was performed and the appearance was compared with first image. All lesions were resected endoscopically, and were subjected to histological assessment and to immunohistochemistry. Serum gastrin levels were determined before and after eradication. RESULTS: Twenty-nine of 37 patients underwent successful eradication. The appearance of 11 lesions (33% of 33 lesions) became indistinct after successful eradication. All lesions were of the superficial-elevated type and the height of the lesions decreased. We detected normal columnar epithelium over the neoplasm in eight of the lesions. Higher expression of single-stranded deoxyribonucleic acid in the deep area was characteristic in tumours with an indistinct appearance. These changes did not correlate with the serum gastrin levels. CONCLUSIONS: The morphology of the gastric neoplasm change after eradication in the short-term. This may contribute to the decreased tumour discovery rate.

Retinoic acid up-regulates erythropoietin production in hepatoma cells and in vitamin A-depleted rats.

Retinoic acid (RA) stimulated the production of erythropoietin (Epo) in a human hepatoma cell line, HepG2 cells. The stimulation was due to the accumulation of Epo mRNA. The Epo production in HepG2 cells was also dependent on O2 tension for cell culture but the enhancement of Epo production by RA was independent of O2 tension, indicating that RA exerts its effect through a pathway different from O2. Oral administration of RA to the vitamin A-depleted rats elevated the concentration of Epo in serum. These results suggest that RA up-regulates EPO production in vivo as well as in vitro.

Purification and characterization of monomeric isocitrate dehydrogenase with NADP(+)-specificity from Vibrio parahaemolyticus Y-4.

NADP(+)-dependent isocitrate dehydrogenase [IDH: EC 1.1.1.42] was purified to electrophoretic homogeneity from Vibrio parahaemolyticus Y-4, and shown to be a monomeric protein of molecular weight 80,000 with a pI of 5.0. The amino acid composition and partial sequence at the N-terminus resembled those reported for other bacterial monomeric IDHs. Immunotitration with antisera to the monomeric and dimeric enzymes (antisera to IDH-II and -I of Vibrio ABE-1) showed an immunochemical distinction between the monomeric and dimeric IDHs, but there is similarity within the IDHs of each group. The circular dichroism spectra of the native and heat-denatured enzyme are also similar to those of monomeric IDH (IDH-II of Vibrio ABE-1). These monomeric IDHs are proteins comprising 17-22% helix and 25-35% beta-pleated sheet in the native state.

Isozymes of isocitrate dehydrogenase from an obligately psychrophilic bacterium, Vibrio sp. strain ABE-1: purification, and modulation of activities by growth conditions.

Each of the two isozymes, which are different in thermostability and quaternary structure, of isocitrate dehydrogenase (NADP+) [IDH: EC 1.1.1.42] was purified to an electrophoretically homogeneous state from an obligately psychrophilic marine bacterium, Vibrio sp. strain ABE-1. Hydrophobic chromatography was an efficient procedure to separate the two isozymes from each other. The isoelectric points of isozyme I (IDH-I; a dimer, Mr 88,100) and isozyme II (IDH-II; a monomer, Mr 80,500) were found to be pH 4.9 and 5.2, respectively. The two isozymes were similar in amino acid compositions, though there were slight differences in the contents of nonpolar and hydroxyl amino acids. However, their NH2-terminal amino acid sequences and immunochemical properties were clearly different from each other. The NH2-terminal amino acid sequence analysis also indicated that the subunits of IDH-I are chemically identical or highly homologous. Non-immuno-crossreactivity between the isozymes enabled us to measure the intracellular contents of the isozymes. IDH-I and -II were found to be differentially regulated in vivo by various growth conditions. IDH-I was induced by acetate, while IDH-II remained almost unchanged.

Erythropoietin gene expression by hydrogen peroxide.

We examined the effect of H2O2 on the regulation of the human erythropoietin (Epo) gene through the GATA sequence in the Epo promoter in Hep3B cells. The addition of exogenous H2O2 in Hep3B cells inhibited hypoxia-induced Epo production of mRNA as assessed by competitive polymerase chain reaction (PCR) and protein by EIA. Likewise, menadione, which increased production of endogenous H2O2, inhibited hypoxia-induced Epo production in Hep3B cells. Furthermore, the expression of the human GATA (hGATA) transcription factor was increased dose-dependently by the addition of H2O2 or menadione in Hep3B cells as assessed by gel mobility shift assay. We concluded that H2O2 inhibits Epo gene expression, because it enhanced the expression of the hGATA transcription factor. Our findings support the hypothesis that GATA is a transcription factor for the Epo gene acting through the oxygen sensor.

Philadelphia chromosome-negative cells with trisomy 8 after busulfan and interferon treatment of Ph1-positive chronic myelogenous leukemia.

A 48-year-old Japanese man with Philadelphia chromosome (Ph1)-positive chronic myelogenous leukemia (CML) was treated with busulfan followed by interferon-alpha (IFN-alpha). Ten months after IFN-alpha treatment, Ph1(-) cells with trisomy 8 were detected by the conventional banding technique and fluorescence in situ hybridization (FISH) analysis. Add(Y)(q12) was also found in Ph1(-) cells with trisomy 8. Although Ph1(+) cells disappeared after the treatment with IFN-alpha, Ph1(-) cells with trisomy 8 did not. We summarize four previous case reports of Ph1(+) CML developing Ph1(-) cells with trisomy 8. All four patients had received busulfan and IFN-alpha. These drugs may be related to the ontogenesis of Ph1(-) cells with trisomy 8, but the significance of Ph1(-) cells with trisomy 8 is not known, and further observation is needed.

All-trans-retinoic acid treatment for chemotherapy-resistant acute adult T-cell leukemia.

We report a case in which treatment with all-trans-retinoic acid (ATRA) improved the clinical features of a 47-year-old female patient with acute adult T-cell leukemia (ATL). The patient was first treated several times with combination chemotherapy. but the number of ATL cells increased and other clinical manifestations progressed. ATRA 60 mg was then administered daily. ATRA treatment dramatically improved the patient's clinical features. In vitro examination revealed that ATRA inhibited the growth of ATL cells from the patient. These findings suggest that ATRA may be a useful treatment for patients with chemotherapy-resistant acute ATL.

Successful treatment of multiple myeloma--associated amyloidosis by interferon-alpha, dimethyl sulfoxide, and VAD (vincristine, adriamycin, and dexamethasone).

A 45-year-old Japanese man was admitted to our hospital because of a mass in the submandibular region, abnormal hematologic findings, and proteinuria. A diagnosis of multiple myeloma was made based on the results of bone marrow analysis and M-protein in hematologic tests, and a diagnosis of amyloidosis was made on the basis of deposition of amyloid in the rectal submucosal and lip tissues and the mass in the submandibular region. Combination therapy of interferon (IFN)-alpha at 1-day intervals and daily oral dimethyl sulfoxide (DMSO) and vincristine, adriamycin, and dexamethasone (VAD) resulted in a marked decrease in the size of the mass and hypertrophy of the back, as well as a decrease in the levels of plasma cells in bone marrow and of M-protein and immunoglobulin G in serum. The results of this case indicate that long-term administration of IFN-alpha and DMSO with VAD is effective in treating amyloidosis with multiple myeloma.

Therapy-related leukemia with a novel 21q22 rearrangement.

We present a case of a 59-year-old Japanese man with therapy-related acute myeloblastic leukemia (AML) after the chemotherapy for non-Hodgkin's lymphoma (NHL). Accumulated doses of cyclophosphamide, procarbazine, doxorubicin, mitoxantrone, and etoposide were 18,300 mg, 3000 mg, 580 mg, 100 mg, and 4150 mg, respectively, which had been administered for the treatment of NHL. Myeloblasts in the peripheral blood increased 43 months after the onset of NHL. He was diagnosed as having AML (M2; FAB classification). The karyotype of the bone marrow cells in the present case contained the following abnormalities: t(2;21)(q21;q22), t(8;21)(q22;q22), and add(13)(q34). In the present case, 645 base pairs of chimeric mRNA were detected by reverse transcription-polymerase chain reaction, indicating the presence of AML1/MTG8 rearrangement. Translocation (2;21)(q21;q22) has not been described previously to our knowledge. It is interesting that the breakpoint of 21q22 existed both in t(2;21) and t(8;21). The disrupted AML1 gene resulting from two 21q22 rearrangements may be involved in the pathogenesis of AML in the present case. The clinical importance of therapy-related AML having the 21q22 rearrangement remains to be examined.

Autologous peripheral blood stem cell transplantation for adults with B-lineage acute lymphoblastic leukemia: a pilot study.

We conducted a pilot study on autologous peripheral blood stem cell transplantation (PBSCT) for 11 adults with B-lineage acute lymphoblastic leukemia (ALL) in first complete remission (CR) or even in those with more advanced stages. All patients achieved CR by induction therapy, of whom 10 were treated with anthracycline, vincristine and prednisolone-based regimens. After consolidation therapy, all patients except one received high-dose cytarabine followed by granulocyte colony-stimulating factor (G-CSF) administration to collect PBSCs. Ten patients received busulfan 4 mg/kg for 4 days, etoposide 20 mg/kg for 3 days and ranimustine 200 mg/m2 for 2 days as a conditioning regimen. One received a regimen consisting of etoposide, cyclophosphamide and total body irradiation. From day 1, G-CSF was given intravenously, and no additional chemotherapy was administered. At the median follow-up time of 30.8 months, four of six patients with standard-risk B-lineage ALL survived within the range of 19.7 to 85.4 months without relapse. In contrast, only one of five with high-risk B-lineage ALL survived for 36.3 months without relapse. Autologous PBSCT as post-remission therapy may prolong CR in adults with standard-risk B-lineage ALL.

B-cell lymphoma terminating in acute monoblastic leukemia.

A case of CD11b-, CD14- and CD36-positive B-cell lymphoma terminating in acute monoblastic leukemia is presented. The patient was initially suspected as having angiotrophic lymphoma due to proliferation of lymphoma cells within the sinusoid of the liver and clinical signs. Following chemotherapy, the lymphoma cells were converted into CD11b- and CD36-positive monoblasts. Lineage switching of B-cell lymphoma to acute monoblastic leukemia may have occurred in this case.

[Staphylococcus aureus sepsis in patients with hematological malignancies: increase in MRSA sepsis].

From January 1978 to August 1990, Staphylococcus aureus bacteremia (SAB) were identified in 31 patients with hematological malignancies at Jichi Medical School hospital. Mortality due to SAB was 48.4% (15/31). Of the variables analyzed, four factors were significantly associated with a poor prognosis; elderly age (p = 0.015), high granulocyte count (more than 500/microliters) (p = 0.015), presence of DIC (p = 0.011) and presence of pneumonia (p = 0.023). The incidence of methicillin-resistant SAB was 32.3% (10/31) and the first patient developed in 1985. Although not statistically significant, there was a trend of higher mortality for methicillin-resistant SAB (70%) than for methicillin-sensitive SAB (38.1%). Most strains of methicillin-resistant Staphylococcus aureus were sensitive to minocycline, chloramphenicol and vancomycin.

[Multiple myeloma with hemolytic anemia and thrombocytopenia].

We report a 59 year old female patient who was diagnosed as having IgG kappa myeloma with hemolytic anemia and thrombocytopenia simultaneously. Although M-protein was suspected to contribute to the hemolysis, the IgG purified from the patient's serum did not bind to red blood cells. Therefore, massive non-specific binding of M-protein to blood cells might contribute to high levels of red blood cell-associated IgG and platelet-associated IgG in the patient.

[A case of acute promyelocytic leukemia (APL) with myeloblastoma in the oral cavity developing after receiving all-trans retinoic acid (ATRA)].

A 44-year-old woman was diagnosed as having acute promyelocytic leukemia (APL) in April 1988. On her first admission, chromosomal translocation (15; 17), +8, and +12 was detected. When she was readmitted to our hospital with the second relapse in May 1990, t(3; 13) and +8 was detected, instead of t(15;17). Complete remission was re-achieved with VP-16, MIT, and BHAC, but the third relapse occurred in September 1990. After obtaining informed consent, she was given etretinate 40 mg per day orally for 17 days, without any effect on leukemia. She was then given all-trans retinoic acid (ATRA) 60 mg per day orally for 29 days. Although a mild granulocytic recovery was observed, no sufficient hematological recovery was obtained (minor response). Besides common side effects of ATRA, such as dry skin and hypertriglycedemia, she had a myeloblastoma in the oral cavity, but it is unknown whether the symptom was a complication of ATRA therapy or not.

[Systemic lupus erythematosus presenting as fulminant thrombotic thrombocytopenic purpura].

A 20-year-old woman visited a nearby hospital because of sudden, severe, and unusual genital bleeding. She also exhibited severe anemia and thrombocytopenia. In transit to our hospital, the patient suddenly suffered cardiac arrest and died soon thereafter despite immediate blood transfusion and therapeutic intubation. Thrombotic thrombocytopenic purpura (TTP) was initially diagnosed at autopsy due to the observation of numerous fragmented erythrocytes in peripheral blood, evidence of hemolysis, and thrombotic microangiopathy in multiple organs. In addition, histopathologic and serologic findings disclosed an association with systemic lupus erythematosus (SLE). Test for anticardiolipin antibody was positive, and hemophagocytic findings were detected in lymph node specimens. Reports of TTP in association with SLE have been increasing in recent years. However, the mechanisms correlating these two illnesses have not been identified. We speculated that the rapid clinical course in this case was attributable to TTP that had been provoked by endothelial microangiopathy due to SLE, and moreover, the fact that the patient's general condition had been seriously complicated by excessive menstrual bleeding.

[Acute undifferentiated leukemia from the viewpoints of diagnosis and therapy].

Myeloperoxidase (MPO)- and Sudan Black B-not more than 3%-positive, esterase staining-negative, lymphoid, megakaryocyte lineage and erythroid surface marker-negative and electron microscopic platelet peroxidase-negative acute leukemia (AL) was diagnosed as acute undifferentiated leukemia (AUL), and myeloid marker (CD13, CD33), electron microscopic MPO (EMMPO), and DNA analysis of immunoglobulin heavy chain and T cell receptor as well as chemotherapy and its reactivity were examined. Of 239 cases of AL, 10 (4.2%) were AUL, and of these 10 cases, 9 were CD13 or CD33-positive AML-MO (MO) cases. Of 9 cases examined for EMMPO, 4 (44%) were positive, and of 3 cases of MO subjected to DNA analysis, 1 and 1 showed rearrangements of immunoglobulin heavy chain and T cell receptor beta chain, respectively. Of 6 cases of MO on myeloid induction therapy, 1 and 1 showed complete remission (CR) and partial remission (PR), respectively, each having lymphoid genotype, and 4 showed no remission (NR), being 3 of them EMMPO-positive. Of 2 cases on lymphoid induction therapy, 1 and 1 showed CR and NR, respectively, the former being EMMPO-positive MO. BHAC-EM therapy with behenoyl cytosine arabinoside, VP-16 and mitoxantrone performed on 2 cases refractory to any one of both these myeloid and lymphoid induction therapies led to CR in all these 2 cases.

[Correlation of risk area and reverse redistribution of 99mTc-sestamibi SPECT in acute myocardial infarction following direct PTCA].

Redistribution of 99mTc-sestamibi is negligible in usual circumstances, but recent reports demonstrated reverse redistribution is detectable in acute myocardial patients. Correlation of risk area, observed in 99mTc-sestamibi "freezed" SPECT image at onset, and delayed images at 5-25 days after onset (post-PTCA image) is evaluated in 19 acute myocardial infarction patients treated with direct PTCA. Reverse redistribution was observed in 85% of reperfused area. Linear relationship of % uptake in each SPECT segment between onset and post-PTCA images (taken at 0.5, 4, and 6 hours after injection) is evaluated and the relationship improves over time course. The correlation coefficient between onset and 6 hours-delayed image is 0.88, and the visual concordance shows 77% of score matching. Delayed 99mTc-sestamibi SPECT image on reperfused AMI seems to represent risk area with some underestimation. It may be useful to estimate both risk and salvaged areas on early and delayed SPECT with a single 99mTc-sestamibi injection.