An EcoRI RFLP downstream of the human c-myc gene.
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Asian People↗
Biomedical subjects
A Tojo
Publications and source records attributed to A Tojo.
A possible correlation between the type of bcr-abl hybrid messenger RNA and platelet count in Philadelphia-positive chronic myelogenous leukemia.
The Philadelphia (Ph1) chromosome, in which the hybrid bcr-abl gene is formed, is thought to be the initial event in chronic myelogenous leukemia (CML). The position of the breakpoint within the breakpoint cluster region (bcr) on Ph1 chromosome and the splicing pattern determine the species of the fused bcr-abl messenger RNA (mRNA). We tried to detect the two types of fused mRNAs in 57 chronic-phase cases of Ph1-positive CML using the polymerase chain reaction procedure (RT-PCR). The bcr exon 2/abl exon 2 fused mRNA (b2-a2) was detected in 17 patients, the bcr exon 3/abl exon 2 fused mRNA (b3-a2) was detected in 34 patients, and both types of mRNA were detected in six patients. The platelet counts of patients who expressed b3-a2 mRNA or both types were significantly higher than those of patients who expressed only b2-a2 (841.5 v 373.5 x 10(9)/L; P less than .015), although there was no significant difference in the white blood cell counts or hemoglobin. This finding suggests a possibility that the type of bcr-abl mRNA may affect the thrombopoietic activity in CML.
Base Sequence↗
Several new approaches to improvement of alpha interferon therapy in chronic myelogenous leukaemia.
Several basic experimental and clinical studies were carried out in an attempt to improve the efficacy of alpha interferon therapy for chronic myelogenous leukaemia (CML). First, the combined use of hydroxyurea (HU) and interferon (500-1000 mg daily) in interferon-resistant cases facilitated maintenance of reduced leucocyte production, or a reduction in the dose of interferon, although suppression of Philadelphia chromosome (Ph1)-positive clones was not observed in most cases. In order to try and decrease the rate of lymphoblastic crisis during the course of interferon therapy, we recently added methotrexate (MTX) (10-15 mg, weekly) to the treatment protocol. Since then, no lymphoblastic crisis has been observed. Second, the in vitro expression of alpha interferon-stimulated gene (ISG) mRNA was shown to be markedly decreased in granulocytes of one representative interferon-resistant case, compared to that in granulocytes of the three interferon-sensitive cases. Interestingly, it was found that the transcriptional activity in this case became almost normal when the blood granulocytes were controlled by the addition of HU. These findings suggest that the in vitro transcriptional assay of ISG mRNA may be clinically useful for predicting alpha interferon efficacy. Third, when genetically manipulated, alpha interferon-producing NIH/3T3 cells were co-transplanted using diffusion chambers into nude mice bearing a CML cell line, KU812, the CML tumour growth was shown to be markedly suppressed. This experimental model for alpha interferon replacement gene therapy suggests some directions for future studies on interferon therapy.
Animals↗
Unregulated expression of the erythropoietin receptor gene caused by insertion of spleen focus-forming virus long terminal repeat in a murine erythroleukemia cell line.
A murine erythroleukemia (MEL) cell line, F5-5, expressed 10,000 binding sites for erythropoietin (EPO) per cell, 10-fold more than was expressed by other murine erythroleukemia cell lines and normal erythroid progenitors. Northern (RNA) and Southern blot analyses revealed overexpression of mRNA for the EPO receptor (EPOR) and rearrangement of one of the EPOR gene alleles in F5-5 cells, respectively. Molecular cloning of F5-5-derived cDNA encoding EPOR revealed that the 5' noncoding region of the EPOR cDNA corresponds to the 3' long terminal repeat sequence of the polycythemic strain of Friend spleen focus-forming virus (F-SFFVP). The aberrant EPOR transcripts containing the 3' long terminal repeat sequence were mainly expressed in F5-5 cells. The same integration upstream of the EPOR gene was also observed in other subclones and the parent cell line. It is possible that overexpression of EPOR by viral promoter insertion will confer growth advantage to an F-SFFVP-infected erythroid progenitor cell, leading to positive clonal selection through further leukemogenic steps.
Amino Acid Sequence↗
Angiotensin II receptor subtypes in rat early proximal tubule.
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Angiotensin II↗
Renal arteriolar diameters in spontaneously hypertensive rats. Vascular cast study.
The relation between the arteriolar diameters and hypertensive glomerulosclerosis was studied by using microvascular casts and histological evaluation. Spontaneously hypertensive rats 4 weeks of age were divided into three groups: nontreated, captopril (40 mg/kg/day)-treated, and trichlormethiazide (1 mg/kg/day) with hydralazine (20 mg/kg/day)-treated. Wistar-Kyoto rats served as controls. At 6 weeks old, the captopril-treated rats showed a lower blood pressure and a larger afferent arteriolar diameter compared with the control rats. At 20 weeks old, the nontreated group exhibited hypertension and a lower arteriolar diameter ratio (afferent to efferent, 0.89 versus 1.22 in control group) because of afferent constriction and efferent dilatation, seen equally in the outer and inner cortexes. Glomerulosclerosis was accentuated only in the inner cortex of the nontreated group (score, 63 versus 29 in control group). In the two treated rat groups, the blood pressure was reduced and arteriolar diameter ratios were similar to those in the control group (1.18 and 1.26). The sclerosis score in the trichlormethiazide with hydralazine-treated rats (score, 26) was lower than in the nontreated rats but not the captopril-treated rats (score, 36). These results indicated that 1) in the hypertensive rats, despite a reduced diameter ratio, glomerulosclerosis was more severe in the inner cortex; 2) two therapies reduced blood pressure and reversed the arteriolar changes, but a decrease in glomerulosclerosis was seen only in the trichlormethiazide with hydralazine-treated rats; and 3) for development of glomerulosclerosis, factors other than hemodynamics may be important in addition to intraglomerular pressure.
Aging↗
Immunohistochemical localization of physiologic urinary proteins in Wistar rats.
Rat physiologic urinary proteins were immunohistochemically localized. In male Wistar rats, urinary protein antigens were present in both hepatic and epithelial cells of the salivary ducts, the coagulating gland, and the prostate gland. In female rats, urinary protein antigens were present in the same proportion of the salivary glands as in males and in the uterine glands, but to a lesser extent than in salivary glands. The results of this study indicate multiple origins of rat urinary proteins. It remains to be determined if female uterine glands contribute to urinary proteins.
Animals↗
[Molecular biology of erythropoietin].
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Amino Acid Sequence↗
Affinity purification of human granulocyte macrophage colony-stimulating factor receptor alpha-chain. Demonstration of binding by photoaffinity labeling.
The human granulocyte macrophage colony-stimulating factor (GM-CSF) receptor alpha-chain, a low affinity component of the receptor, was solubilized and affinity-purified from human placenta using biotinylated GM-CSF. Scatchard analysis of 125I-GM-CSF binding to the placental membrane extract disclosed that the GM-CSF receptor had a dissociation constant (Kd) of 0.5-0.8 nM, corresponding to the Kd value of the GM-CSF receptor alpha-chain on the intact placental membrane. Affinity labeling of the solubilized protein using a photoreactive cross-linking agent, N-hydroxysuccinimidyl-4-azidobenzoate (HSAB), demonstrated a single specific band of 70-95 kDa representing a ligand-receptor complex. Approximately 2 g of the placental membrane extract was subjected to a biotinylated GM-CSF-fixed streptavidin-agarose column, resulting in a single major band at 70 kDa on a silver-stained sodium dodecyl sulfate gel. The radioiodination for the purified material disclosed that the purified protein had an approximate molecular mass of 70 kDa and a pI of 6.6. Binding activity of the purified material was demonstrated by photoaffinity labeling using HSAB-125I-GM-CSF, producing a similar specific band at 70-95 kDa as was demonstrated for the crude protein.
Affinity Labels↗
Isolation of genes highly expressed in early and late stages of Friend virus-induced erythroleukemia in mice.
We isolated two genes that are highly expressed in Friend erythroleukemia. The first one, expressed at an extremely high level in a late-stage tumorigenic cell line F5-5, was found to be a species of intracisternal A-particle. Most of those genomes in F5-5 DNA were not amplified nor rearranged compared with those in the normal cell DNA, suggesting transcriptional activation. Another late-stage cell line T3K showed similar results. The second gene, highly expressed in early-stage cells but weaker in late-stage cells, was identified as one of the heme-synthesizing enzymes, uroporphyrinogen decarboxylase. Usefulness of these two genes for the analysis of erythroid transformation and differentiation will be discussed.
Animals↗
Morphometric analysis of arteriolar diameters in experimental nephropathies: application of microvascular casts.
Renal arteriolar diameters were measured, using microvascular resin casts, in two hyperfiltration models of rats: the remnants kidney of subtotal nephrectomy (NX) and streptozotocin-induced diabetic kidney (DM). In the NX, the blood pressure was elevated, urinary protein excretion was markedly increased and glomeruli were severely damaged. In the DM, although the blood pressure remained normal, urinary protein excretion was significantly increased and glomeruli were damaged but to a lesser extent than in the NX group. In the NX group, the afferent arteriole was dilated and the efferent arteriole was constricted. In the DM group, the afferent arteriole was dilated, while the efferent arteriole remained unchanged. The results showed that afferent arteriolar dilatation was seen in both the NX and DM groups, possibly leading to the glomerular damage. In the NX group, the systemic high blood pressure and efferent arteriolar constriction augmented glomerular damage significantly.
Animals↗
Variations in renal arteriolar diameter in deoxycorticosterone acetate-salt hypertensive rats. A microvascular cast study.
The relation between hypertensive glomerular damage and arteriolar diameter was examined in a microvascular cast study in deoxycorticosterone acetate (DOCA)-salt hypertensive rats. The blood pressure and urinary protein excretion increased progressively in the DOCA rats. In controls afferent arteriolar diameters increased during the course of the experiment, and efferent arteriolar diameters remained unchanged. In the DOCA rats, however, afferent arteriolar diameters did not change significantly, while efferent arteriolar diameters increased. Histological studies showed severe arteriolosclerosis and glomerulosclerosis in the DOCA rats. The results show that these arteriolar changes might contribute to the reduction of glomerular capillary pressure in the development of DOCA-salt hypertension. However, they are not sufficient to protect glomeruli from hypertensive damage.
Animals↗
Oligomerization of Friend spleen focus-forming virus (SFFV) env glycoproteins.
The glycoprotein gp55 and its processed form gp65, which are encoded by the env gene of Friend spleen focus-forming virus (SFFV), have been implicated in the initiation of the murine acute erythroleukemia induced by Friend virus (FV). Analyses of these glycoproteins by chemical crosslinking and nonreducing/reducing two-dimensional electrophoresis showed that both gp55 and gp65 exist as monomer and disulfide-bonded dimer and trimer. These oligomers could be detected in various FV-infected erythroleukemia cell lines, as well as in the spleen cells of FV-induced erythroleukemic mice, suggesting that oligomerization is an intrinsic feature of SFFV env glycoproteins.
Animals↗
Identification and cellular distribution of distinct proteins forming human GM-CSF receptor.
Two proteins forming the receptor for human granulocyte-macrophage colony-stimulating factor (GM-CSF)1 were identified and characterized. One with apparent Mr of about 80,000 was defined as alpha-chain and has Kd of 0.7-2.8 nM. The other binding molecule with apparent Mr of about 135,000 was defined as beta-chain and is related to the high-affinity binding with Kd of 10-40 pM. The binding kinetic studies confirmed that the 125I-GM-CSF associated slower to and dissociated more rapidly from the alpha-chain than the beta-chain. The alpha-chain is expressed not only on hemopoietic cells but also on full-term placental tissues, choriocarcinoma cells, and other solid tumor cells. In contrast, the distribution of the beta-chain is restricted on hemopoietic cells. The alpha-chain probably corresponds to the low-affinity GM-CSF receptor whose cDNA has been cloned and sequenced.
Granulocyte-Macrophage Colony-Stimulating Factor↗
Philadelphia chromosome positive B-cell type malignant lymphoma expressing an aberrant 190 kDa bcr-abl protein.
The Philadelphia (Ph) chromosome translocation, t(9:22) (q34;q11) is found in some acute lymphoid leukaemias (ALL) and acute myeloid leukaemias (AML). Although cytogenetically all pH chromosomes appear similar, the 22q11 breakpoints found in acute leukaemias are of two kinds, those within the major breakpoint cluster region (Mbcr-1) of the BCR gene as found in chronic myelogenous leukaemia (CML), and those within the first intron of this gene. In the former group the molecular events are the same as those found in CML, p210 bcr-abl, encoded by 8.5 kb mRNA; however, a new aberrant protein, p190 bcr-abl, is found in the latter group. Ph translocation is also found in a few cases with malignant lymphoma, but it has not been characterized at the molecular level. We describe here a non-Hodgkin's lymphoma case with primary splenic presentation, which showed a complex Ph translocation. Neoplastic cells were of a B-cell origin (HLA-DR+, sIgM+, sIg lambda +, CALLA-). Molecular studies revealed the expression of p190 bcr-abl with no Mbcr-1 rearrangement. Our case indicates that the same Ph translocation as seen in acute leukaemias can be found in haematologic disorders other than leukaemias, suggesting that a c-abl gene activating mechanism may be involved in the pathogenesis of wide spectrum of haematologic malignancies.
Adult↗
Effects of sairei-to on the relapse of steroid-dependent nephrotic syndrome.
Sairei-To (Chai-Ling-Tang) was administered to four patients with steroid-dependent relapsing nephrotic syndrome. It was associated with prednisolone and immunosuppressive agents. Histological diagnosis was minimal change in three patients and mild focal glomerulonephritis in one patient. After the start of Sairei-To administration the relapse was markedly suppressed in three patients but not at all in the other. Although we could consider Sairei-To effective for steroid-dependent nephrotic syndrome in the present study, a larger study is necessary to confirm its efficacy.
Adult↗
Effects of a Japanese medicinal plant on the rat subtotal nephrectomy model: evaluation of its effect by microvascular casts.
Effects of a Japanese medicinal plant named Sairei-To, were examined in a rat experimental renal disease. Thirteen weeks after subtotal nephrectomy, the blood pressures in rats given Sairei-To (Sairei-To rats) were lower than those without Sairei-To. The urinary protein excretions and glomerular sclerosis were markedly decreased in the Sairei-To treated rats. Arteriolar diameters were measured using microvascular casts. The afferent and efferent arterioles were both significantly dilatated. The efferent arterioles in Sairei-To rats were dilated to a greater extent than that of the afferent. These results indicated that Sairei-To lessened renal damages in the rat subtotal nephrectomy model, possibly through the blood pressure reduction and the efferent arteriolar dilatation.
Animals↗
Effects of atrial natriuretic peptide on renal arterioles: morphometric analysis using microvascular casts.
In normal rat kidneys, the effect of atrial natriuretic peptide (ANP) on the diameter of the arterioles was evaluated by scanning electron microscopy of vascular casts. Acryl resin was infused into rat kidneys during the administration of ANP, either alone or with norepinephrine (NE). ANP infusion constricted the proximal efferent arteriole in the superficial cortex. Although NE constricted the proximal and distal segments of the afferent arteriole in the superficial cortex, the addition of ANP reversed the constriction and further constricted the efferent arteriole. In the deep cortex, only the proximal segment of the afferent arteriole was dilated by ANP when infused with NE. In a separate set of experiments, ANP increased both the glomerular filtration rate (GFR) and urinary sodium excretion (UNaV), and NE decreased the renal blood flow (RBF). However, administration of ANP after NE recovered RBF and increased GFR as well as UNaV. Results indicate that ANP increases GFR and natriuresis by constricting the efferent arteriole. NE appears to decrease RBF by constricting the afferent arteriole. ANP antagonizes the renal effects of NE primarily by dilating afferent arterioles.
Animals↗