RESUMEN
The pathogenesis of cyclophosphamide (CY)-induced cardiotoxicity remains unknown, and methods for its prevention have not been established. To elucidate the acute structural changes that take place in myocardial cells and the pathways leading to myocardial damage under high-dose CY treatments, we performed detailed pathological analyses of myocardial tissue obtained from C57BL/6J mice subjected to a high-dose CY treatment. Additionally, we analysed the genome-wide cardiomyocyte expression profiles of mice subjected to the high-dose CY treatment. Treatment with CY (400 mg/kg/day intraperitoneally for two days) caused marked ultrastructural aberrations, as observed using electron microscopy, although these aberrations could not be observed using optical microscopy. The expansion of the transverse tubule and sarcoplasmic reticulum, turbulence in myocardial fibre travel, and a low contractile protein density were observed in cardiomyocytes. The high-dose CY treatment altered the cardiomyocyte expression of 1210 genes (with 675 genes upregulated and 535 genes downregulated) associated with cell-cell junctions, inflammatory responses, cardiomyopathy, and cardiac muscle function, as determined using microarray analysis (|Z-score| > 2.0). The expression of functionally important genes related to myocardial contraction and the regulation of calcium ion levels was validated using real-time polymerase chain reaction analysis. The results of the gene expression profiling, functional annotation clustering, and Kyoto Encyclopedia of Genes and Genomes pathway functional-classification analysis suggest that CY-induced cardiotoxicity is associated with the disruption of the Ca2+ signalling pathway.
RESUMEN
This study examined the prognostic impact of erythroblast predominance (EP) in 61 patients with myelodysplastic syndromes (MDS) (n = 51) or acute myeloid leukemia (n = 10) treated with azacitidine. Median age was 78 years. EP, defined as > 40% erythroblasts and M/E < 1.0, was found in 21 patients, including 9 complex karyotypes (CK). In the 24 CK of the entire cohort, 5 were hyperdiploid and 15 were monosomal karyotype with -5/5q-, and 10 had immunophenotypically CD41/cyCD41 positive blasts (cyCD41+). The complete response (CR) rate was 32.8%. Median follow-up was 14 months, and median overall survival (OS) was 17 months. Although all patients with EP achieved high CR rates (61.9%) and extended OS (28 M, P = 0.056), patients with EP and cyCD41+ blasts had shorter OS (8 M, P = 0.002). EP (HR 0.39, P = 0.009) and cyCD41+ (HR 3.49, P = 0.018) were identified as prognostic factors in multivariate analysis. All patients with cyCD41+ had hyperdiploid or CK with -5/5q-. In conclusion, we divided patients into three risk categories: high (cyCD41+), low (EP without cyCD41+), and intermediate (non-CD41+ and non-EP), and median OS in these categories was 34, 17 and 8 months, respectively (P < 0.001).
Asunto(s)
Leucemia Mieloide Aguda , Síndromes Mielodisplásicos , Cariotipo Anormal , Anciano , Antimetabolitos Antineoplásicos/uso terapéutico , Azacitidina/uso terapéutico , Eritroblastos , Humanos , Leucemia Mieloide Aguda/tratamiento farmacológico , Leucemia Mieloide Aguda/genética , Síndromes Mielodisplásicos/tratamiento farmacológico , PronósticoRESUMEN
Hermansky-Pudlak syndrome type 2 (HPS2) is an extremely rare autosomal recessive inherited disease characterized by partial oculocutaneous albinism (OCA), bleeding diathesis due to a storage pool deficiency and immunodeficiency. The disorder is caused by disruption of the adapter protein 3 complex, which is involved in impaired intracellular vesicle transport. Here, we report the first case of a 1-year-old girl with HPS2 in Asia. She had no specific symptoms other than OCA and neutropenia. We analyzed her platelet function using transmission electron microscopy and a platelet aggregation test, cytotoxic degranulation assay of her natural killer (NK) cells and bleeding time, the results of which led to the diagnosis of HPS2. Although her NK-cell cytotoxic degranulation was impaired, she had not developed signs of hemophagocytic lymphohistiocytosis (HLH) or fibrosing lung disease. Molecular genetic analyses showed novel heterozygous mutations (c.188T>A [p.M63K] and c.2546>A [p.L849X]) in AP3B1. When examining patients with OCA, blood tests should be performed to confirm neutrophil count, bleeding time and platelet agglutination. When HPS2 is suspected, detailed immunological tests should be considered, and attention should be paid to HLH and pulmonary lesions immediately and over the long term.
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Complejo 3 de Proteína Adaptadora/genética , Subunidades beta de Complejo de Proteína Adaptadora/genética , Síndrome de Hermanski-Pudlak/genética , Plaquetas/patología , Plaquetas/ultraestructura , Análisis Mutacional de ADN , Femenino , Cabello/patología , Cabello/ultraestructura , Síndrome de Hermanski-Pudlak/sangre , Síndrome de Hermanski-Pudlak/diagnóstico , Síndrome de Hermanski-Pudlak/patología , Heterocigoto , Humanos , Lactante , Japón , Microscopía Electrónica de Transmisión , MutaciónRESUMEN
The RUNX1-EVI1 gene generated by the t(3;21) translocation encodes a chimeric transcription factor and is a causative gene in the development of de novo acute megakaryoblastic leukemia and leukemic transformation of hematopoietic stem cell tumors. Heterozygous RUNX1-EVI1 knock-in mice die in utero due to hemorrhage in the central nervous system and spinal cord and complete abolishment of definitive hematopoiesis in the fetal liver. On the other hand, the chimeric knock-in mouse develops acute megakaryoblastic leukemia. We created another mouse model of RUNX1-EVI1 using transplantation of retrovirus-infected bone marrow cells. Some mice transplanted with RUNX1-EVI1-expressing bone marrow cells developed acute megakaryoblastic leukemia within eight months, and the other non-leukemic mice showed thrombocytosis at around a year. In the non-leukemic mice, dysplastic megakaryocytes proliferated in the bone marrow and frequently infiltrated into the spleen, which was not associated with marrow fibrosis. In the leukemic mice, their tumor cells were positive for c-kit and CD41, and negative for TER119. Although they were negative for platelet peroxidase in the electron microscopic analysis, they had multiple centrioles in the cytoplasm, which are characteristic of megakaryocytes that undergo endomitosis. The leukemic cells were serially transplantable, and gene-expression analyses using quantitative RT-PCR arrays revealed that they showed significantly elevated expression of stem cell, primitive hematopoietic cell and endothelial cell-related genes compared with normal bone marrow cells. All these data suggested that RUNX1-EVI1 caused dysplastic hematopoiesis or leukemia of the megakaryocytic lineage and endowed gene expression profiles distinctive of immature hematopoietic cells.
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Subunidad alfa 2 del Factor de Unión al Sitio Principal/biosíntesis , Hematopoyesis , Leucemia/metabolismo , Proteína del Locus del Complejo MDS1 y EV11/biosíntesis , Megacariocitos/metabolismo , Proteínas de Fusión Oncogénica/biosíntesis , Enfermedad Aguda , Aloinjertos , Animales , Trasplante de Médula Ósea , Subunidad alfa 2 del Factor de Unión al Sitio Principal/genética , Leucemia/genética , Leucemia/patología , Proteína del Locus del Complejo MDS1 y EV11/genética , Megacariocitos/patología , Ratones , Ratones Transgénicos , Proteínas de Fusión Oncogénica/genéticaAsunto(s)
Aberraciones Cromosómicas , Cromosomas Humanos Par 3 , Leucemia Basofílica Aguda/genética , Células de la Médula Ósea/patología , Células de la Médula Ósea/ultraestructura , Gránulos Citoplasmáticos/patología , Gránulos Citoplasmáticos/ultraestructura , Diagnóstico Diferencial , Eritema/etiología , Fiebre/etiología , Humanos , Leucemia Basofílica Aguda/diagnóstico , Leucemia Basofílica Aguda/patología , Leucemia Basofílica Aguda/fisiopatología , Masculino , Microscopía Electrónica de Transmisión , Persona de Mediana EdadRESUMEN
To maintain a trusting relationship between clinical and hospital laboratory staff, highly reliable reporting based on precise quality control of the test results is required. Testing work is divided into 3 steps: pre-testing, testing, and post-testing. Quality control (QC) of laboratory testing has been performed to improve the precision and accuracy of measurements after sample collection, mainly in the testing step. However, various factors influencing the measurement results are present in the process from requests for testing to the reporting of the test results, and it is necessary to make efforts to minimize these factors. The characteristics of POCT devices and reagents are their simple operation method, compact size, and use at sites other than laboratories, and most users are physicians and nurses. Sample measurement rooms have opened at sites other than medical institutions, and testing using POCT-compatible devices and reagents has been rapidly spreading. It is very important to clarify factors leading to false high and low values in the pre-testing step. The results of investigating reasons for predicted events were presented, and the necessity of quality management in the pre-testing step was clarified. If the pre-testing step is not properly performed, accuracy cannot be assured even though quality management of the testing and post-testing steps is optimal.
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Ciencia del Laboratorio Clínico , Pruebas en el Punto de Atención , Garantía de la Calidad de Atención de Salud , Control de Calidad , Recolección de Muestras de Sangre/métodos , Cromatografía de Afinidad , Humanos , Ciencia del Laboratorio Clínico/métodos , Manejo de EspecímenesRESUMEN
Devices for self monitoring of blood glucose (SMBG) are used by diabetic patients themselves. Evaluation of the environmental temperature during use by patients showed the importance of the temperature environment. When measurement is performed at a temperature outside the measurement temperature range, a function to display errors or does not display measurement values is indispensable. Measures to reduce puncture pain in SMBG include use of a thin needle, shallow puncturing, and the selection of the palm or forearm instead of the fingertip as a puncture site. Using these measures to reduce puncture pain, the blood volume required for measurement is often difficult to collect. Therefore, the puncture site is massaged or squeezed. However, these actions were reported to have serious influences on measurement values. Changes in the blood glucose and Ht levels and the hemolysis phenomenon were confirmed as the influences of the massaging or squeezing method. The correct blood collection method free of such influences is blood sampling from the fingertip using the pushing out method.
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Automonitorización de la Glucosa Sanguínea/métodos , Recolección de Muestras de Sangre/métodos , Humanos , TemperaturaRESUMEN
The Great East Japan Earthquake caused a tragic tsunami and resulted in serious damage to north region of Japan on March 11, 2011. The Japanese Society of Laboratory Medicine, JSLM launched an ad hoc Committee to support Laboratory Medicine affairs in the affected area. We expected that laboratory testing demands would increase during the weeks following the disaster. We decided to support the use of Point-of-Care Testing. Many POCT devices use battery-powered analyzers. This is definite advantage for their use in areas with limited access to power and water supplies. We contacted many companies about the possibility of providing POCT devices, IVD reagents and/or any laboratory supplies including disposable materials. Finally, forty companies agreed to support this project and we received list of reagents materials for more than one hundred IVD tests. We entered this information on our web site and continued to update it as additional support was received. Once a request of support was received, communication were made to confirm the amount of material, the method of shipping/receipt and if any specific training that would be required for its use at the testing site. Also, we dispatched volunteer Medical Technologists for eight weeks to assist in the laboratory work. Some of the crucial points in recruiting volunteer laboratory professions are expenses and accommodations. We prepared not only accommodations but also transportation methods and covered all expenses including insurance and meals. Our relief activities have shown that Laboratory Medicine and Medical Technologists are useful in disaster-affected area.
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Planificación en Desastres/organización & administración , Terremotos , Ciencia del Laboratorio Clínico/organización & administración , Sociedades Científicas/organización & administración , Humanos , Japón , Personal de Laboratorio Clínico , Sistemas de Atención de PuntoRESUMEN
The AML1/EVI1 chimeric gene is created by the t(3;21)(q26;q22) chromosomal translocation seen in patients with leukemic transformation of myelodysplastic syndrome or blastic crisis of chronic myelogenous leukemia. We knocked-in the AML1/EVI1 chimeric gene into mouse Aml1 genomic locus to explore its effect in developmental hematopoiesis in vivo. AML1/EVI1/+ embryo showed defective hematopoiesis in the fetal liver and died around embryonic day 13.5 (E13.5) as a result of hemorrhage in the central nervous system. The peripheral blood had yolk-sac-derived nucleated erythroblasts but lacked erythrocytes of the definitive origin. Although E12.5 fetal liver contained progenitors for macrophage only, E13.5 fetal liver contained multilineage progenitors capable of differentiating into dysplastic myelocyte and megakaryocyte. No erythroid progenitor was detected in E12.5 or E13.5 fetal liver. Hematopoietic progenitors from E13.5 AML1/EVI1/+ fetal liver were highly capable of self-renewal compared with those from wild-type liver. Maintained expression of PU.1 gene and decreased expression of LMO2 and SCL genes may explain the aberrant hematopoiesis in AML1/EVI1/+ fetal liver.
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Hematopoyesis/genética , Proteínas de Fusión Oncogénica/fisiología , Proteínas Adaptadoras Transductoras de Señales , Animales , Subunidad alfa 2 del Factor de Unión al Sitio Principal , Proteínas de Unión al ADN/genética , Embrión de Mamíferos , Eritroblastos , Células Madre Hematopoyéticas/citología , Hemorragias Intracraneales/etiología , Proteínas con Dominio LIM , Hígado/citología , Hígado/embriología , Proteína del Locus del Complejo MDS1 y EV11 , Metaloproteínas/genética , Ratones , Ratones Transgénicos , Células Madre Multipotentes , Proteínas de Fusión Oncogénica/genética , Proteínas Proto-Oncogénicas/genética , Proto-Oncogenes/genética , Transactivadores/genética , Factores de Transcripción/genéticaRESUMEN
Embryonic development of multilineage hematopoiesis requires the precisely regulated expression of lineage-specific transcription factors, including AML-1 (encoded by Runx1; also known as CBFA-2 or PEBP-2alphaB). In vitro studies and findings in human diseases, including leukemias, myelodysplastic syndromes and familial platelet disorder with predisposition to acute myeloid leukemia (AML), suggest that AML-1 has a pivotal role in adult hematopoiesis. However, this role has not been fully uncovered in vivo because of the embryonic lethality of Runx1 knockout in mice. Here we assess the requirement of AML-1/Runx1 in adult hematopoiesis using an inducible gene-targeting method. In the absence of AML-1, hematopoietic progenitors were fully maintained with normal myeloid cell development. However, AML-1-deficient bone marrow showed inhibition of megakaryocytic maturation, increased hematopoietic progenitor cells and defective T- and B-lymphocyte development. AML-1 is thus required for maturation of megakaryocytes and differentiation of T and B cells, but not for maintenance of hematopoietic stem cells (HSCs) in adult hematopoiesis.
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Diferenciación Celular/fisiología , Proteínas de Unión al ADN/metabolismo , Hematopoyesis/fisiología , Células Madre Hematopoyéticas/fisiología , Linfocitos/fisiología , Megacariocitos/fisiología , Proteínas Proto-Oncogénicas/metabolismo , Factores de Transcripción/metabolismo , Adulto , Animales , Células de la Médula Ósea/citología , Células de la Médula Ósea/metabolismo , Linaje de la Célula , Tamaño de la Célula , Subunidad alfa 2 del Factor de Unión al Sitio Principal , Proteínas de Unión al ADN/genética , Marcación de Gen/métodos , Células Madre Hematopoyéticas/citología , Humanos , Linfocitos/citología , Megacariocitos/citología , Ratones , Ratones Endogámicos , Ratones Transgénicos , Proteínas Proto-Oncogénicas/genética , Factores de Transcripción/genéticaRESUMEN
Frequent development of renal cell carcinomas in hybrids between Japanese toads (Bufo japonicus) and imported Chinese toads (Bufo raddei) was first reported by 2 of our authors in 1987. Such renal tumors of toads had never been observed previously in the laboratory. To confirm the observation and to establish a new animal model system, hybrids between female Japanese and male Chinese toads were newly generated from 3 pairs of parents and pathological changes in their kidneys were examined sequentially over 6 years. In hybrids from 2 of the 3 pairs, bilateral polycystic kidney developed at a high frequency from 3 months after fertilization, this being associated with the emergence of atypical, premalignant-appearing cells in proximal tubules. Papillary lesions developed after 12 months and renal cell carcinomas after 48 months. Such pathological changes were never seen in non-hybrid Chinese or Japanese toads. Electron microscopy showed no evidence of any viral participation. This unique toad model may prove useful for investigation of the underlying mechanisms of genetically determined renal cell carcinogenesis.
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Bufonidae , Carcinoma de Células Renales/veterinaria , Quimera , Neoplasias Renales/veterinaria , Enfermedades Renales Poliquísticas/veterinaria , Animales , Carcinoma de Células Renales/patología , China , Cruzamientos Genéticos , Modelos Animales de Enfermedad , Femenino , Japón , Neoplasias Renales/patología , Lectinas/química , Masculino , Enfermedades Renales Poliquísticas/patologíaRESUMEN
BACKGROUND: Abnormal iron deposition may cause oxidant-induced damage in various organs. We have previously reported that continuous administration of angiotensin II to rats results in an overt iron deposition in the renal tubular epithelial cells, which may have a role in angiotensin II-induced renal damage. In the present study, we investigated the role of iron in the development of cardiac injury induced by angiotensin II. METHODS AND RESULTS: Angiotensin II was continuously infused to rats at a dose of 0.7 mg/kg per day for 7 consecutive days. No iron deposits were observed in the hearts of untreated rats, whereas iron deposition was seen in the cells in the subepicardial and granulation regions after angiotensin II infusion. Concomitant administration of deferoxamine, an iron chelator, significantly reduced the extent of cardiac fibrosis, which suggests that iron deposition aggravates the cardiac fibrosis induced by angiotensin II. Iron overload caused by the administration of iron-dextran resulted in an augmentation of cardiac fibrosis and the generation of neointimal cells in the coronary artery in angiotensin II-infused rats. By contrast, neointima was not formed in the cardiac vessels in norepinephrine-infused rats with iron overload. CONCLUSIONS: Cardiac iron deposition may be involved in the development of cardiac fibrosis induced by angiotensin II. In addition, iron overload may enhance the formation of neointima under conditions of increased circulating angiotensin II but not catecholamines.
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Angiotensina II , Fibrosis/inducido químicamente , Corazón/efectos de los fármacos , Sobrecarga de Hierro/inducido químicamente , Miocardio/patología , Túnica Íntima/efectos de los fármacos , Antagonistas de Receptores de Angiotensina , Animales , Presión Sanguínea/efectos de los fármacos , Western Blotting , Modelos Animales de Enfermedad , Sinergismo Farmacológico , Fibrosis/patología , Fibrosis/prevención & control , Hemo Oxigenasa (Desciclizante)/biosíntesis , Hemo-Oxigenasa 1 , Hipertensión/inducido químicamente , Hipertensión/complicaciones , Inmunohistoquímica , Quelantes del Hierro/farmacología , Sobrecarga de Hierro/patología , Complejo Hierro-Dextran , Losartán/farmacología , Masculino , Miocardio/ultraestructura , Norepinefrina , Ratas , Ratas Sprague-Dawley , Receptor de Angiotensina Tipo 1 , Túnica Íntima/patología , Vasodilatadores/farmacologíaRESUMEN
Acute experimental iron loading causes iron to accumulate in the renal tissue. The accumulation of iron may play a role in enhancing oxidant-induced tubular injury by producing increased amounts of reactive oxygen species. From findings in cells from heme oxygenase-1 (HO-1)-deficient mice, HO-1 is postulated to prevent abnormal intracellular iron accumulation. Recently, it has been reported that HO-1 is induced in the renal tubular epithelial cells, in which iron is deposited after iron loading, and that this HO-1 induction may be involved in ameliorating iron-induced renal toxicity. We previously showed that chronic administration of angiotensin II to rats induces HO-1 expression in the tubular epithelial cells. These observations led us to investigate whether there is a link between iron deposition and HO-1 induction in renal tubular cells in rats undergoing angiotensin II infusion. In the present study, rats were given angiotensin II for continuously 7 days. Prussian blue staining revealed the distinct deposits of iron in the proximal tubular epithelial cells after angiotensin II administration. Electron microscopy demonstrated that iron particles were present in the lysosomes of these cells. Histologic and immunohistochemical analyses showed that stainable iron and immunoreactive ferritin and HO-1 were colocalized in the tubular epithelial cells. Treatment of angiotensin II-infused rats with an iron chelator, deferoxamine, blocked the abnormal iron deposition in kidneys and also the induced expression of HO-1 and ferritin expression. Furthermore, deferoxamine treatment suppressed the angiotensin II-induced increase in the urinary excretion of protein and N-acetyl-beta-D-glucosaminidase, a marker of tubular injury; however, deferoxamine did not affect the angiotensin II-induced decrease in glomerular filtration rate. These results suggest that angiotensin II causes renal injury, in part, by inducing the deposition of iron in the kidney.