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1.
Cell Death Dis ; 5: e1132, 2014 Mar 20.
Article in English | MEDLINE | ID: mdl-24651435

ABSTRACT

Expression of apoptotic protease activating factor-1 (Apaf-1) gradually decreases during brain development, and this decrease is likely responsible for the decreased sensitivity of brain tissue to apoptosis. However, the mechanism by which Apaf-1 expression is decreased remains elusive. In the present study, we found that four microRNAs (miR-23a/b and miR-27a/b) of miR-23a-27a-24 and miR-23b-27b-24 clusters play key roles in modulating the expression of Apaf-1. First, we found that miR-23a/b and miR-27a/b suppressed the expression of Apaf-1 in vitro. Interestingly, the expression of the miR-23-27-24 clusters in the mouse cortex gradually increased in a manner that was inversely correlated with the pattern of Apaf-1 expression. Second, hypoxic injuries during fetal distress caused reduced expression of the miR-23b and miR-27b that was inversely correlated with an elevation of Apaf-1 expression during neuronal apoptosis. Third, we made neuronal-specific transgenic mice and found that overexpressing the miR-23b and miR-27b in mouse neurons inhibited the neuronal apoptosis induced by intrauterine hypoxia. In conclusion, our results demonstrate, in central neural system, that miR-23a/b and miR-27a/b are endogenous inhibitory factors of Apaf-1 expression and regulate the sensitivity of neurons to apoptosis. Our findings may also have implications for the potential target role of microRNAs in the treatment of neuronal apoptosis-related diseases.


Subject(s)
Apoptosis , Apoptotic Protease-Activating Factor 1/metabolism , Cerebral Cortex/metabolism , Fetal Hypoxia/metabolism , MicroRNAs/metabolism , Neurons/metabolism , Animals , Animals, Newborn , Apoptotic Protease-Activating Factor 1/genetics , Cell Hypoxia , Cerebral Cortex/embryology , Cerebral Cortex/pathology , Disease Models, Animal , Female , Fetal Hypoxia/genetics , Fetal Hypoxia/pathology , Gene Expression Regulation, Developmental , Gestational Age , HEK293 Cells , Humans , Mice , Mice, Inbred C57BL , Mice, Transgenic , MicroRNAs/genetics , Neurons/pathology , Pregnancy , Signal Transduction , Time Factors , Transfection
2.
Oncogene ; 28(32): 2910-8, 2009 Aug 13.
Article in English | MEDLINE | ID: mdl-19503097

ABSTRACT

The partition-defective 3 (PAR-3) protein is implicated in the formation of tight junctions at epithelial cell-cell contacts. We investigated DNA copy number aberrations in human esophageal squamous cell carcinoma (ESCC) cell lines using a high-density oligonucleotide microarray and found a homozygous deletion of PARD3 (the gene encoding PAR-3). Exogenous expression of PARD3 in ESCC cells lacking this gene enhanced the recruitment of zonula occludens 1 (ZO-1), a marker of tight junctions, to sites of cell-cell contact. Conversely, knockdown of PARD3 in ESCC cells expressing this gene caused a disruption of ZO-1 localization at cell-cell borders. A copy number loss of PARD3 was observed in 15% of primary ESCC cells. Expression of PARD3 was significantly reduced in primary ESCC tumors compared with their nontumorous counterparts, and this reduced expression was associated with positive lymph node metastasis and poor differentiation. Our results suggest that deletion and reduced expression of PARD3 may be a novel mechanism that drives the progression of ESCC.


Subject(s)
Carcinoma, Squamous Cell/genetics , Cell Cycle Proteins/genetics , Esophageal Neoplasms/genetics , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Membrane Proteins/genetics , Adaptor Proteins, Signal Transducing , Aged , Carcinoma, Squamous Cell/metabolism , Carcinoma, Squamous Cell/pathology , Cell Cycle Proteins/metabolism , Cell Line, Tumor , Cell Movement , Esophageal Neoplasms/metabolism , Esophageal Neoplasms/pathology , Gene Deletion , Gene Dosage , Homozygote , Humans , Immunoblotting , Infant , Intercellular Junctions/metabolism , Male , Membrane Proteins/metabolism , Microscopy, Confocal , Microscopy, Fluorescence , Middle Aged , Oligonucleotide Array Sequence Analysis , Phosphoproteins/metabolism , RNA Interference , Reverse Transcriptase Polymerase Chain Reaction , Zonula Occludens-1 Protein
3.
Thorax ; 64(4): 365-7, 2009 Apr.
Article in English | MEDLINE | ID: mdl-19329736

ABSTRACT

A case of pulmonary alveolar microlithiasis occurring in an inbred family is presented. A genome-wide analysis of the patient's genomic DNA using a high-density single nucleotide polymorphism (SNP) array revealed a small intragenetic mutation at SLC34A2. The results suggest that the high-density SNP array has the power to identify a recessive disease gene(s) even in the analysis of only a single inbred patient.


Subject(s)
Calculi/genetics , Lung Diseases/genetics , Polymorphism, Single Nucleotide/genetics , Pulmonary Alveoli , Sequence Deletion/genetics , Female , Genome, Human , Humans , Middle Aged
4.
Oncogene ; 28(10): 1385-92, 2009 Mar 12.
Article in English | MEDLINE | ID: mdl-19137007

ABSTRACT

Dysregulated expression of microRNAs (miRNAs) is associated with a variety of diseases, including colorectal cancer. By comparing more than 200 miRNAs in 13 pairs of matched colorectal cancer and normal adjacent tissue samples through qRT-PCR and microarray analysis, we found a widespread disruption of miRNA expression during colorectal tumorigenesis. In particular, among a panel of presumed targets generated by in silico analysis that may interact with these aberrantly expressed miRNAs, KRAS oncogene has been further experimentally validated as the target of miR-143. First, an inverse correlation between KRAS protein and miR-143 in vivo was found. Second, KRAS expression in Lovo cells was significantly abolished by treatment with miR-143 mimic, whereas miR-143 inhibitor increased KRAS protein level. Third, luciferase reporter assay confirmed that miR-143 directly recognize the 3'-untranslated region of KRAS transcripts. Four, Lovo cells treated with miR-143 inhibitor showed a stimulated cell proliferation, whereas miR-143 overexpression had an opposite effect. Finally, inhibition of KRAS expression by miR-143 inhibits constitutive phosphorylation of ERK1/2. Taken together, the present study provides the first evidences that miR-143 is significant in suppressing colorectal cancer cell growth through inhibition of KRAS translation.


Subject(s)
Colorectal Neoplasms/prevention & control , MicroRNAs/physiology , Proto-Oncogene Proteins/antagonists & inhibitors , ras Proteins/antagonists & inhibitors , Colorectal Neoplasms/genetics , Genes, Tumor Suppressor , Humans , MicroRNAs/analysis , Oligonucleotide Array Sequence Analysis , Proto-Oncogene Proteins p21(ras) , Reverse Transcriptase Polymerase Chain Reaction
7.
Bone Marrow Transplant ; 36(7): 639-47, 2005 Oct.
Article in English | MEDLINE | ID: mdl-16044136

ABSTRACT

Chronic graft-versus-host disease (cGVHD) occurs in approximately 60-80% of those who survive over 100 days after allogeneic hematopoietic stem cell transplantation (allo-HSCT). However, the pathophysiology of cGVHD is poorly understood. To gain more insight into the immunological mechanism of cGVHD, we examine cytokine production of peripheral blood T cells from 19 patients in the chronic phase of allo-HSCT. The percentage of IFN-gamma-producing CD8(+) T cells among CD8(+) T cells was significantly higher in patients with or without cGVHD than in normal control subjects (P<0.001). On the other hand, the percentage of IL-4-producing CD8(+) T cells among CD8(+) T cells was significantly higher in patients with cGVHD (mean 3.3%; range 1.3-8.2%) than in patients without cGVHD (mean 1.2%; range 0.8-1.7%) and normal control subjects (mean 1.1%; range 0.1-1.6%) (both P<0.001). By contrast, the percentage of IL-4-producing CD4(+) T cells was not different among patients with and without cGVHD and normal controls. These findings suggest that IL-4-producing CD8(+) T cells may be an immunological marker of cGVHD.


Subject(s)
CD8-Positive T-Lymphocytes/metabolism , Graft vs Host Disease/immunology , Interleukin-4/biosynthesis , Adolescent , Adult , Case-Control Studies , Chronic Disease , Cytokines/biosynthesis , Cytokines/metabolism , Female , Flow Cytometry , Hematopoietic Stem Cell Transplantation/methods , Humans , Interferon-gamma/metabolism , Interleukin-4/metabolism , Male , Middle Aged , Recurrence , Time Factors , Transplantation, Homologous/methods
8.
Bone Marrow Transplant ; 33(12): 1231-3, 2004 Jun.
Article in English | MEDLINE | ID: mdl-15094752

ABSTRACT

We treated three patients with steroid-refractory acute graft-versus-host disease (aGVHD) with intra-arterial steroid-injection therapy (IAST). Two patients with gut aGVHD received IAST into both superior and inferior mesenteric arteries, while one patient with liver aGVHD received IAST into the proper hepatic artery. The volume of stools and the bilirubin level improved soon after IAST. Angiography of the superior and inferior mesenteric arteries was performed in the two patients with steroid-refractory gut aGVHD, and identical abnormal findings were obtained. IAST might be an earlier option for steroid-refractory aGVHD.


Subject(s)
Drug Resistance , Graft vs Host Disease/drug therapy , Steroids/administration & dosage , Acute Disease , Adult , Angiography , Female , Hematologic Diseases/complications , Hematologic Diseases/therapy , Hematopoietic Stem Cell Transplantation/adverse effects , Humans , Injections, Intra-Arterial , Intestinal Diseases/drug therapy , Liver Diseases/drug therapy , Male , Middle Aged
9.
Bone Marrow Transplant ; 33(10): 989-96, 2004 May.
Article in English | MEDLINE | ID: mdl-15064686

ABSTRACT

The recipients of allogeneic hematopoietic stem cell transplantation (allo-HSCT) often develop acute graft-versus-host disease (aGVHD), which is closely related to morbidity and mortality. However, the essential part of the immune responses elicited in aGVHD remains largely unknown. We attempt to determine if peripheral blood dendritic cells (PBDCs) are altered in aGVHD, and find that the number of PBDCs (both myeloid and lymphoid DCs) is significantly decreased. Immunohistochemical staining of the biopsied skin from patients with aGVHD demonstrates that a number of fascin(+) cells with dendritic projections infiltrate the dermis of the skin. Based on these findings, we hypothesize that the PBDCs are recruited to the affected tissues and may thus play important roles in immune responses elicited in aGVHD.


Subject(s)
Dendritic Cells/metabolism , Graft vs Host Disease/blood , Leukemia/therapy , Acute Disease , Adolescent , Adult , Aged , Biopsy , Carrier Proteins/metabolism , Culture Media/pharmacology , Female , Hematopoietic Stem Cell Transplantation/methods , Humans , Immunohistochemistry , Leukocytes, Mononuclear/metabolism , Lymphocytes/metabolism , Male , Microfilament Proteins/metabolism , Middle Aged , Myeloid Cells/metabolism , Skin/metabolism , Time Factors
10.
Ann Nucl Med ; 15(3): 271-6, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11545201

ABSTRACT

A 73-year-old man with aortic regurgitation was examined by 123I-alpha-methyl-p-iodophenylpentadecanoic acid (BMIPP) myocardial single photon emission computed tomography (SPECT) in 1995. Myocardial accumulation was not evident on either the early or the delayed image obtained 15 minutes and 3 hours, respectively, after injecting 123I-BMIPP. Flow cytometric analysis of CD36 expression in monocytes and platelets identified a type I CD36 deficiency. The patient was hospitalized for severe heart failure in 1999. Upon admission, the cardiothoracic ratio on chest X-rays was 73%, and the left ventricular end-diastolic diameter on echocardiograms was enlarged to 77 mm. On the second day, we performed 123I-BMIPP myocardial SPECT. Myocardial accumulation was evident in the delayed, but not in the early image. We repeated 123I-BMIPP myocardial SPECT on the 10th day after admission. Myocardial accumulation was evident on both early and delayed images. 99mTc-tetrofosmin myocardial SPECT was immediately performed after 123I-BMIPP myocardial SPECT to distinguish myocardial from pooling images in the left ventricle, but, because the images from both 99Tc-tetrofosmin and 123I-BMIPP myocardial SPECT were idential, we considered that the 123I-BMIPP myocardial SPECT images reflected the actual myocardial condition. The CD36 molecule transports long-chain fatty acid (LCFA) on the myocardial membrane, but 123I-BMIPP scintigraphy does not show any myocardial accumulation in patients with type I CD36 deficiency, indicating that myocardial LCFA uptake occurs through CD36 on the human myocardial membrane. Even though our patient had type I CD36 deficiency, BMIPP was uptaken by the myocardium during heart failure, suggesting a variant pathway on the human myocardial membrane for LCFA uptake.


Subject(s)
Aortic Valve Insufficiency/diagnostic imaging , CD36 Antigens/genetics , Fatty Acids/pharmacokinetics , Heart/diagnostic imaging , Iodine Radioisotopes/pharmacokinetics , Iodobenzenes/pharmacokinetics , Aged , Antigens, CD/blood , Antigens, CD/genetics , CD36 Antigens/blood , Follow-Up Studies , Heart Failure/diagnostic imaging , Humans , Male , Time Factors , Tissue Distribution , Tomography, Emission-Computed, Single-Photon
11.
Biochemistry ; 40(35): 10491-9, 2001 Sep 04.
Article in English | MEDLINE | ID: mdl-11523990

ABSTRACT

Helices IV and V in the lactose permease of Escherichia coli contain the major determinants for substrate binding [Glu126 (helix IV), Arg144 (helix V), and Cys148 (helix V)]. Structural and dynamic features of this region were studied by using site-directed sulfhydryl modification of 48 single-Cys replacement mutants with N-[(14)C]ethylmaleimide (NEM) in the absence or presence of ligand. In right-side-out membrane vesicles, Cys residues in the cytoplasmic halves of both helices react with NEM in the absence of ligand, while Cys residues in the periplasmic halves do not. Five Cys replacement mutants at the periplasmic end of helix V and one at the cytoplasmic end of helix V label only in the presence of ligand. Interestingly, in addition to native Cys148, a known binding-site residue, labeling of mutant Ala122 --> Cys, which is located in helix IV across from Cys148, is markedly attenuated by ligand. Furthermore, alkylation of the Ala122 --> Cys mutant blocks transport, and protection is afforded by substrate, indicating that Ala122 is also a component of the sugar binding site. Methanethiosulfonate ethylsulfonate, an impermeant thiol reagent shown clearly in this paper to be impermeant in E. coli spheroplasts, was used to identify substituted Cys side chains exposed to water and accessible from the periplasmic side. Most of the Cys mutants in the cytoplasmic halves of helices IV and V, as well as two residues in the intervening loop, are accessible to the aqueous phase from the periplasmic face of the membrane. The findings indicate that the cytoplasmic halves of helices IV and V are more reactive/accessible to thiol reagents and more exposed to solvent than the periplasmic half. Furthermore, positions that exhibit ligand-induced changes are located for the most part in the vicinity of the residues directly involved in substrate binding, as well as the cytoplasmic loop between helices IV and V.


Subject(s)
Escherichia coli Proteins , Escherichia coli/enzymology , Membrane Transport Proteins/chemistry , Monosaccharide Transport Proteins , Symporters , Affinity Labels , Amino Acid Sequence , Binding Sites , Cytoplasm/enzymology , Escherichia coli/genetics , Membrane Transport Proteins/genetics , Membrane Transport Proteins/metabolism , Models, Molecular , Molecular Sequence Data , Point Mutation , Protein Conformation , Substrate Specificity , Sulfhydryl Compounds
13.
J Cardiol ; 37(4): 215-22, 2001 Apr.
Article in Japanese | MEDLINE | ID: mdl-11337931

ABSTRACT

Iodine-123 15-(p-iodophenyl)-3-R,S-methylpentadecanoic acid myocardial single photon emission computed tomography(iodine-123 BMIPP myocardial SPECT) is useful for the diagnosis and evaluation of ischemic myocardial disease or cardiomyopathy. Changes in fatty acid metabolism in the post-myocardial ischemic state were evaluated using iodine-123 BMIPP myocardial SPECT. A 77-year-old woman was hospitalized for treatment of acute myocardial infarction. Emergency coronary angiography showed severe stenosis with delayed filling of contrast medium in the middle portion of the left anterior descending artery, so primary percutaneous transluminal coronary angioplasty was performed successfully. On the second day, iodine-123 BMIPP uptake was decreased slightly on the early imaging in the apico-anterior region, and increased slightly on the delayed imaging. On the seventh day, iodine-123 BMIPP uptake was decreased moderately or markedly in the apico-anterior region on the early imaging, and decreased markedly on the delayed imaging. Iodine-123 BMIPP myocardial SPECT subsequently became almost normalized. These unusual dynamic changes In iodine-123 BMIPP myocardial SPECT imaging may reflect metabolic changes of fatty acids in the ischemic state, the size of the triacylglycerol pool, and the degree of turnover in the triacylglycerol pool.


Subject(s)
Fatty Acids , Iodine Radioisotopes , Iodobenzenes , Myocardial Infarction/diagnostic imaging , Radiopharmaceuticals , Tomography, Emission-Computed, Single-Photon , Aged , Female , Humans
14.
Ann Nucl Med ; 14(4): 303-9, 2000 Aug.
Article in English | MEDLINE | ID: mdl-11023032

ABSTRACT

A 55-yr-old man underwent surgery. Soon after the procedure was finished, the patient complained of chest pain, and the electrocardiogram showed increase in the ST-segment in some leads. Emergency angiography showed normal coronary arteries, but there was asynergy in the left ventricle, and delayed filling of contrast medium was observed in the LCA. An intracoronary infusion of isosorbide dinitrate did not improve the delayed filling of contrast medium or ST segment increase in the electrocardiogram. Soon after nicorandil was injected into the LCA, the patient's symptoms, electrocardiogram, and delayed filling of contrast medium dramatically improved. On the second day, initial imaging by 123I-BMIPP myocardial SPECT showed a moderate increase in tracer uptake in the apico-anteroseptal region and a moderate decrease in tracer uptake in the lateral region, in which the first left ventriculography showed akinesis, and delayed imaging revealed a moderate increase in tracer uptake in the apical region and a high washout of 123I-BMIPP in the anteroseptal and lateral regions. On the sixth day, initial imaging by 123I-BMIPP myocardial SPECT showed a moderate decrease in tracer uptake in the apical and lateral regions and a mild decrease in tracer uptake in the anteroseptal region, and delayed imaging revealed a moderate increase in tracer uptake in the apical region and a high washout of 123I-BMIPP in the anteroseptal and lateral regions. By the 30th day, 123I-BMIPP myocardial SPECT had normalized. We consider that these dynamic changes in 123I-BMIPP myocardial SPECT imaging may reflect metabolic changes in fatty acids in the ischemic state, the size of the triacylglycerol pool, and the degree of turnover in the triacylglycerol pool.


Subject(s)
Coronary Vasospasm/complications , Fatty Acids , Iodine Radioisotopes , Iodobenzenes , Myocardial Stunning/diagnostic imaging , Myocardial Stunning/etiology , Tomography, Emission-Computed, Single-Photon/methods , Electrocardiography , Humans , Male , Middle Aged , Myocardial Stunning/metabolism , Organophosphorus Compounds , Organotechnetium Compounds , Technetium Tc 99m Pyrophosphate , Triglycerides/metabolism
15.
Kaku Igaku ; 37(4): 327-32, 2000 Jul.
Article in Japanese | MEDLINE | ID: mdl-10965653

ABSTRACT

This study was designed to evaluate the clinical usefulness of 123I-BMIPP myocardial SPECT for detecting cardiac involvement in patients with collagen disease. We studied 12 patients with systemic lupus erythematosus (SLE), 8 with progressive systemic sclerosis (PSS), 6 with polymyositis/dermatomyositis (PM/DM) and 3 with allergic granulomatosis and angiitis (AGA). A 111 MBq of 123I-BMIPP was intravenously injected at rest, and SPECT images were obtained at 15 min after the injection. Seven of 12 SLE, 6 of 8 PSS, 3 of 6 PM/DM and all 3 AGA patients showed an abnormal tracer uptake. The left ventricular ejection fraction was inversely correlated with a BMIPP abnormality. The regional wall motion abnormality was reduced in regions with reduced tracer uptake. These findings suggest that 123I-BMIPP imaging could be useful for assessment of cardiac involvement in patients with collagen disease.


Subject(s)
Collagen Diseases/diagnostic imaging , Fatty Acids , Heart/diagnostic imaging , Iodine Radioisotopes , Iodobenzenes , Tomography, Emission-Computed, Single-Photon/methods , Aged , Female , Humans , Male , Middle Aged
16.
Kaku Igaku ; 37(3): 209-15, 2000 May.
Article in Japanese | MEDLINE | ID: mdl-10860351

ABSTRACT

This study was designed to evaluate the clinical usefulness of delayed exercise images in 99mTc-tetrofosmin (TF) myocardial SPECT in the diagnosis of vasospastic angina pectoris. We studied 30 patients with vasospastic angina, 10 of 30 patients (group A) had both effort and rest angina, 20 of 30 patients (group B) had rest angina. A 370 MBq of TF was intravenously injected at peak exercise, and initial (EX-I) and delayed exercise (EX-D) images were obtained at 30 min and 180 min after the injection. An additional 740 MBq of TF was intravenously reinjected after EX-D image acquisition, and rest images were obtained 30 min after the reinjection. The left ventricular wall was divided into 9 segments. Regional myocardial uptakes of TF were scored by 4-point defect score (0 = normal, 1 = mildly reduced, 2 = moderately reduced, and 3 = severely reduced). Total defect score (TDS) was calculated from the sum of defect scores in 9 segments. Reverse redistribution (RR) was defined as increase of more than 2 in TDS on EX-D images. In group A, 4 of 10 cases (40%) showed decreased uptake on EX-I images, 6 of 10 cases (60%) revealed RR on EX-D images, and none of the patients showed decreased uptake on rest images. In group B, no one showed decreased uptake on EX-I and rest images, 11 of 20 cases (55%) revealed RR on EX-D images. The mean +/- SD of TDS were 2.9 +/- 3.4, 5.1 +/- 4.5, 0.5 +/- 0.5 on EX-I, EX-D, rest images in group A, and serially 0.4 +/- 0.5, 3.3 +/- 3.6, 0.4 +/- 0.5 in group B. Regional wall motion abnormality was reduced in regions with RR. RR on EX-D images may reflect ischemic damaged but viable myocardium in vasospastic angina. The clinical usefulness of exercise-rest TF imaging in detection of organic coronary artery disease has been well established. Therefore, exercise-rest TF imaging with additional delayed exercise image could evaluate not only organic coronary artery disease but also coronary artery vasospasm.


Subject(s)
Angina Pectoris/diagnostic imaging , Coronary Vasospasm/diagnostic imaging , Heart/diagnostic imaging , Organophosphorus Compounds , Organotechnetium Compounds , Radiopharmaceuticals , Aged , Angina Pectoris/etiology , Coronary Vasospasm/complications , Exercise Test , Female , Humans , Male , Middle Aged , Time Factors , Tomography, Emission-Computed, Single-Photon
17.
Kaku Igaku ; 37(2): 115-20, 2000 Mar.
Article in Japanese | MEDLINE | ID: mdl-10783570

ABSTRACT

Case 1 involved a 62-year-old woman with complete atrioventricular block and case 2 involved a 70-year-old woman with sick sinus syndrome. In both cases, echocardiography, 123I-15-(p-iodophenyl)-3-R,S-methyl pentadecanoic acid (BMIPP) and 99mTc-tetrofosmin myocardial single photon emission computed tomographic (SPECT) findings were normal. Coronary arteriography and left ventriculography also revealed normal in both cases. After pacemaker implantation (DDD-type in case 1 and VVI-type in case 2), both patient's electrocardiograms revealed left bundle branch block-type conduction disturbance, and echocardiography showed asynchronous hypokinesia in the septal region. One year later, although both patient's ATP loading tetrofosmin SPECT were normal, BMIPP SPECT showed reduced uptake and increased washout in the septal region. These findings suggest that these changes of BMIPP SPECT might be caused by left bundle branch block-type conduction disturbance after pacemaker implantation.


Subject(s)
Bundle-Branch Block/etiology , Fatty Acids , Heart/diagnostic imaging , Iodine Radioisotopes , Iodobenzenes , Pacemaker, Artificial/adverse effects , Radiopharmaceuticals , Tomography, Emission-Computed, Single-Photon , Aged , Bundle-Branch Block/diagnostic imaging , Female , Heart Block/diagnostic imaging , Heart Block/therapy , Humans , Middle Aged , Sick Sinus Syndrome/diagnostic imaging , Sick Sinus Syndrome/therapy
18.
Rinsho Ketsueki ; 41(1): 25-31, 2000 Jan.
Article in Japanese | MEDLINE | ID: mdl-10695395

ABSTRACT

In October 1996, a 26-year-old woman was given a diagnosis of acute myeloblastic leukemia, FAB subtype M1. Treatment with combined chemotherapy achieved a complete remission (CR). In May 1997, the patient received an allogenic bone marrow transplant (BMT) from an HLA-identical sibling donor. Cyclosporine (CsA) and short-term methotrexate were given for graft-versus-host disease (GVHD) prophylaxis. Successful engraftment was obtained and signs of acute or chronic GVHD never developed. Five months after BMT, the patient experienced low-grade fever and blurred vision. Retinal examination demonstrated intraretinal hemorrhages, cotton-wool spots, and retinal detachments, which were presumably attributable to multiple thrombosis of retinal microvessels. The patient also exhibited hemolytic anemia with red cell fragmentation, thrombocytopenia, elevated lactate dehydrogenase, and renal impairment, and was thus given a diagnosis of BMT-associated thrombotic microangiopathy (BMT-TM). Discontinuation of CsA and administration of ticlopidine and prednisolone induced successful recovery from BMT-TM. Three months after the onset of BMT-TM, however, the patient experienced generalized clonic-tonic seizures with consciousness loss. Single-photon-emission computed tomography revealed blood-flow disturbances in the brain, suggesting the recurrence of microthrombosis. Accordingly, multiple transfusions of fresh frozen plasma were administered together with dipyridamole and aspirin. The patient gradually recovered and remained asymptomatic through the following 13 months. Currently, early diagnosis of BMT-TM is considered to be difficult. We suggest that careful examination of the ocular base may be useful for the early detection of BMT-TM.


Subject(s)
Bone Marrow Transplantation/adverse effects , Retinal Vessels , Thrombosis/etiology , Vision Disorders/etiology , Adult , Aspirin/therapeutic use , Dipyridamole/therapeutic use , Female , Humans , Leukemia, Myeloid, Acute/therapy , Microcirculation , Plasma , Prednisolone/therapeutic use , Recurrence , Thrombosis/diagnosis , Thrombosis/drug therapy , Treatment Outcome , Vision Disorders/drug therapy
19.
Insect Biochem Mol Biol ; 30(2): 107-17, 2000 Feb.
Article in English | MEDLINE | ID: mdl-10696586

ABSTRACT

Knowledge-based protein modeling and substrate docking experiments as well as structural and sequence comparisons were performed to identify potential active-site residues in chitinase, a molting enzyme from the tobacco hornworm, Munduca sexta. We report here the identification of an active-site amino acid residue, W145. Several mutated forms of the gene encoding this protein were generated by site-directed mutagenesis, expressed in a baculovirus-insect cell-line system, and the corresponding mutant proteins were purified and characterized for their catalytic and substrate-binding properties. W145, which is present in the presumptive catalytic site, was selected for mutation to phenylalanine (F) and glycine (G), and the resulting mutant enzymes were characterized to evaluate the mechanistic role of this residue. The wild-type and W145F mutant proteins exhibited similar hydrolytic activities towards a tri-GlcNAc oligosaccharide substrate, but the former was approximately twofold more active towards a polymeric chitin-modified substrate. The W145G mutant protein was inactive towards both substrates, although it still retained its ability to bind chitin. Therefore, W145 is required for optimal catalytic activity but is not essential for binding to chitin. Measurement of kinetic constants of the wild-type and mutant proteins suggests that W145 increases the affinity of the enzyme for the polymeric substrate and also extends the alkaline pH range in which the enzyme is active.


Subject(s)
Chitinases/chemistry , Chitinases/metabolism , Manduca/enzymology , Amino Acid Sequence , Animals , Base Sequence , Catalysis , Catalytic Domain , Chitinases/genetics , DNA Primers/genetics , Hydrogen-Ion Concentration , Manduca/genetics , Models, Molecular , Mutagenesis, Site-Directed , Oligosaccharides/chemistry , Protein Conformation , Protein Structure, Tertiary , Substrate Specificity , Tryptophan/chemistry
20.
Kaku Igaku ; 37(1): 23-8, 2000 Jan.
Article in Japanese | MEDLINE | ID: mdl-10714064

ABSTRACT

Case 1 involved a 52-year-old man with angina chest pain at rest and case 2 involved a 63-year-old woman with chest oppression at rest. An electrocardiogram (ECG) showed negative T wave in III and aVF leads in case 1, and complete atrioventricular block and ST segment depression in II, III, aVF, and V5-6 leads in case 2. In both cases, 99mTc-tetrofosmin myocardial SPECT showed reduced uptake in the inferior and posterior wall. Although bath patients' left coronary arteriographies were normal, right coronary arteriographies revealed severely delayed filling of contrast medium without significant narrowing of epicardial coronary arteries, suggesting microembolism or microvascular vasospasm. An intracoronary infusion of isosorbide dinitrate did not improve the delayed filling of contrast medium or ST segment depression on ECG. Soon after intracoronary infusion of diltiazem in case 1 and nicorandil in case 2, coronary arterial flows were normalized, chest symptoms disappeared, and ECG findings were normalized. The next day, both patients' 99mTc-tetrofosmin myocardial SPECT showed normal uptake. These findings suggest that myocardial ischemia in these cases might be explained as having been caused by microvascular spasm.


Subject(s)
Heart/diagnostic imaging , Microvascular Angina/complications , Myocardial Ischemia/diagnostic imaging , Organophosphorus Compounds , Organotechnetium Compounds , Radiopharmaceuticals , Tomography, Emission-Computed, Single-Photon , Calcium Channel Blockers/therapeutic use , Diltiazem/therapeutic use , Female , Humans , Male , Middle Aged , Myocardial Ischemia/drug therapy , Myocardial Ischemia/etiology , Nicorandil/therapeutic use , Treatment Outcome , Vasodilator Agents/therapeutic use
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