RESUMEN
Valproic acid (VPA) is a widely used drug for epilepsy. However, precise molecular mechanisms relevant to VPA's side effects remain elusive. This study identifies a VPA-sensitive mutant strain ( vas21 ) in fission yeast with a missense mutation (T256I) in the nucleotide sugar-binding motif of the GDP-mannose transporter Vrg4 . This mutation impairs protein glycosylation, as evidenced by altered acid phosphatase mobility. We also found that Vrg4 overexpression deteriorates cell growth. Our results highlight the role of Vrg4 in glycosylation and implicate impaired glycosylation as a potential mechanism underlying VPA sensitivity. The new allele of vrg4 will be useful in glycobiology and pharmacology.
RESUMEN
Laminin is a family of basement membrane proteins, whose selective and spatiotemporal expression profiles are linked to their various functions in development, maintenance, and functional regulation of different tissues. In the liver, α1-and α5-containing laminin isoforms have been documented to be critically involved in the developmental process of the epithelial tissue of the bile duct. However, possible roles of other laminin isoforms in bile duct formation and function remain elusive. Here, we evaluated public single-cell RNA sequencing databases on human liver cells to reveal expression landscape of laminin genes, and found that genes for laminin-332 subunits were conjointly expressed in the EPCAM+ biliary epithelial cell population. Expression of the ß3 and γ2 subunit genes was restricted to biliary epithelial cells in the liver and, remarkably, showed apparent heterogeneity among them. We confirmed the heterogeneous nature of the laminin-ß3 expression in murine livers, which was firmly related to morphological substructures in the biliary epithelium. Finally, we generated the liver epithelial tissue-specific laminin- ß3 knockout mice and found that this laminin subunit was dispensable under physiological conditions. Together, our present findings have identified the ß3 subunit and the related laminin-332 isoform as useful markers and potentially important regulatory molecules for future understanding of pathophysiology in the hepatobiliary system.
Asunto(s)
Moléculas de Adhesión Celular/análisis , Hígado/metabolismo , Animales , Conductos Biliares/metabolismo , Moléculas de Adhesión Celular/genética , Células Epiteliales/metabolismo , Femenino , Expresión Génica , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , KalininaRESUMEN
Resin glycosides are well known as the purgative ingredients, which are characteristic of convolvulaceous plants. Calystegia hederacea Wall. is a perennial herbaceous vine that is widespread throughout India and East Asia. All parts of this plant are used for the treatment of menoxenia, gonorrhea, etc. Alkaline hydrolysis of the crude resin glycoside fraction of the whole plants of C. hederacea yielded four new glycosidic acids, calyhedic acids A, B, C, and D, along with two known glycosidic acids, calysolic acids A and C, and three known organic acids, 2S-methylbutyric, tiglic, and 2R,3R-nilic acids. Their structures were characterized on the basis of spectroscopic data and chemical evidence. Calyhedic acids A, B, and D were penta-, hexa-, and hepta-glycosides of 12S-hydroxyhexadecanoic acid, respectively, and cayhedic acid C was an isomer of calyhedic acid D, in which the 12S-hydroxyhexadecanoyl residue of calyhedic acid D was replaced by a 11S-hydroxyhexadecanoyl residue. Additionally, cytotoxic activity toward HL-60 human promyelocytic leukemia cells of the crude resin glycoside fraction, the glycosidic acid fraction, calyhedic acid A, and calysolic acid A from C. hederacea was evaluated. Furthermore, to clarify the structure-activity relationship of resin glycosides, the activities of six genuine resin glycosides with calysolic acid A or calysolic acid C as the glycosidic acid, which were isolated from C. soldanella, were examined. Among them, the crude resin glycoside fraction and five genuine resin glycosides with macrolactone structures demonstrated clear cytotoxic activities, while the glycosidic acid fraction, calyhedric acid A, calysolic acid A, and a genuine non-macrolactone-type resin glycoside were either inactive or exhibited weaker activity than the tested macrolactone-type resin glycosides.
Asunto(s)
Calystegia/química , Glicósidos/química , Resinas de Plantas/química , Humanos , Hidrólisis , Estructura Molecular , Oligosacáridos , Plantas Medicinales/química , Relación Estructura-ActividadRESUMEN
The present study aimed to investigate the effects of the combination of Marukome Nenrin miso, which has natriuretic effects, and Marukome MK-34-1 miso, which has potent angiotensin converting enzyme inhibitory effects, on blood pressure (BP) in humans. A total of 40 subjects aged 40-69 years with high-normal BP or stage I hypertension were randomly assigned to two groups: 1) the miso group (32 g 2:1 w/w Nenrin and MK-34-1 with 3.8 g salt/day) or 2) the control soy food group (14.4 g soy food with 0.2 g salt/day). The levels of major nutrients were equal in the miso and control food servings, except for the fiber and Na levels, which were higher in the miso food serving. Daytime and nighttime BP were measured with an automated BP monitor. Compared with the soy food intake, miso intake for 8 weeks did not affect daytime clinical BP but significantly decreased nighttime BP without affecting pulse rate (PR). Moreover, miso shifted the nighttime BP profile to lower levels than those at baseline. Soy food intake did not change the nighttime BP profile after 8 weeks. Miso intake also tended to reduce nighttime BP in a subgroup with stage 1 hypertension compared with the results of the soy food group participants and shifted the nighttime BP profile toward lower levels than those recorded at baseline. Miso intake did not influence lipid or glucose metabolism. In conclusion, this is the first report showing that miso reduces nighttime BP in humans. Miso may do so by shrinking the fluid spaces in the body and/or deactivating the adrenergic nervous system.
Asunto(s)
Inhibidores de la Enzima Convertidora de Angiotensina/administración & dosificación , Hipertensión/dietoterapia , Prehipertensión/dietoterapia , Alimentos de Soja , Adulto , Anciano , Presión Sanguínea , Ritmo Circadiano , Método Doble Ciego , Femenino , Frecuencia Cardíaca , Humanos , Hipertensión/prevención & control , Masculino , Persona de Mediana EdadRESUMEN
We newly manufactured miso rich in angiotensin-converting enzyme (ACE) inhibitory activity (Marukome MK-34-1, shinki miso) and investigated its antihypertensive properties in rat models of genetic hypertension. ACE inhibitory activity was tenfold higher in shinki miso than in commercially available Marukome Nenrin miso (nenrin miso). The inhibitory activity of shinki miso was confined to <3 kDa fractions and was detected in several fractions with high polarity by C18 high-performance liquid chromatography. Systolic blood pressure (SBP) increased age-dependently in stroke-prone spontaneously hypertensive rats (SHRSP/Izm) given a 0.6% (w/v) NaCl solution (salt solution group) that matched the salt content of the miso solutions. This SBP increase was attenuated in both the 5% nenrin and 5% shinki miso solution groups compared to the salt solution group. The reduction in SBP was greater in rats fed shinki than in rats fed nenrin miso. Similarly, in a salt-induced hypertension model with Dahl rats, the 5% nenrin miso solution attenuated the rising SBP observed in the salt solution group. Moreover, combining 5% nenrin miso with 5% shinki miso (2:1, v/v) (awase miso group) significantly decreased the SBP per gram salt intake by 8% compared with the nenrin miso treatment. However, there were no differences in urinary Na excretion between the nenrin and awase miso groups. In conclusion, we produced a new miso with potent ACE inhibitory activity that reduced spontaneous and salt-induced hypertension. These results suggest that salt sensitivity is decreased by the addition of shinki miso to nenrin miso.
Asunto(s)
Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Antihipertensivos/farmacología , Presión Sanguínea/efectos de los fármacos , Hipertensión/tratamiento farmacológico , Hipertensión/genética , Alimentos de Soja , Envejecimiento , Animales , Peso Corporal , Cromatografía Líquida de Alta Presión , Masculino , Tamaño de los Órganos , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Endogámicas Dahl , Ratas Endogámicas SHR , Sodio/orina , Alimentos de Soja/análisis , Accidente Cerebrovascular/genéticaRESUMEN
Under various conditions of liver injury, the intrahepatic biliary epithelium undergoes dynamic tissue expansion and remodeling, a process known as ductular reaction. Mouse models defective in inducing such a tissue-remodeling process are more susceptible to liver injury, suggesting a crucial role of this process in liver regeneration. However, the molecular mechanisms regulating the biliary epithelial cell (BEC) dynamics in the ductular reaction remain largely unclear. Here, we demonstrate that the transcription factor Krüppel-like factor 5 (Klf5) is highly enriched in mouse liver BECs and plays a key role in regulating the ductular reaction, specifically under cholestatic injury conditions. Although mice lacking Klf5 in the entire liver epithelium, including both hepatocytes and BECs (Klf5-LKO (liver epithelial-specific knockout) mice), did not exhibit any apparent phenotype in the hepatobiliary system under normal conditions, they exhibited significant defects in biliary epithelial tissue remodeling upon 3,5-diethoxycarbonyl-1,4-dihydrocollidine-induced cholangitis, concomitantly with exacerbated cholestasis and reduced survival rate. In contrast, mice lacking Klf5 solely in hepatocytes did not exhibit any such phenotypes, confirming Klf5's specific role in BECs. RNA-sequencing analyses of BECs isolated from the Klf5-LKO mouse livers revealed that the Klf5 deficiency primarily affected expression of cell cycle-related genes. Moreover, immunostaining analysis with the proliferation marker Ki67 disclosed that the Klf5-LKO mice had significantly reduced BEC proliferation levels upon injury. These results indicate that Klf5 plays a critical role in the ductular reaction and biliary epithelial tissue expansion and remodeling by inducing BEC proliferation and thereby contributing to liver regeneration.
Asunto(s)
Conductos Biliares Intrahepáticos/metabolismo , Colestasis/metabolismo , Células Epiteliales/metabolismo , Factores de Transcripción de Tipo Kruppel/biosíntesis , Regeneración Hepática , Hígado/metabolismo , Animales , Conductos Biliares Intrahepáticos/patología , Ciclo Celular/efectos de los fármacos , Colestasis/inducido químicamente , Colestasis/genética , Colestasis/patología , Células Epiteliales/patología , Regulación de la Expresión Génica/efectos de los fármacos , Hepatocitos/metabolismo , Hepatocitos/patología , Factores de Transcripción de Tipo Kruppel/genética , Hígado/lesiones , Hígado/patología , Ratones , Ratones Noqueados , Piridinas/toxicidadRESUMEN
Environmental pollution by potentially toxic elements (PTEs) has become a serious problem with increasing industrialization and the disturbance of natural biogeochemical cycles. Jatropha is an oilseed-bearing shrub with high potential for biodiesel production in arid regions. In this study, we examined the physiological responses of this plant to five representative PTEs (Cd, Cr, Cu, Ni, and Zn) in a hydroponic culture. Application of higher concentrations of Cd and Zn led to severe leaf chlorosis, and Cd, Cu, and Ni treatments resulted in significant growth retardation. Higher enrichment of the applied PTEs in the shoots was observed for Zn- and Cd-treated plants, with the latter reaching 24-fold enrichment in plants exposed to 10 µM Cd, suggesting that Jatropha can cope with relatively higher internal concentrations of toxic Cd. Although Cd stress led to the disturbance of essential mineral homeostasis and photosynthesis, this induced an increase in thiol compounds in the roots, suggesting defensive responses of Jatropha to PTEs. This study showed that Jatropha exhibits distinct sensitivities and physiological responses to different PTEs. This study also provides basic knowledge for diagnosing the physiological status of Jatropha trees for potential dual use in afforestation and as a sustainable energy supply.
Asunto(s)
Contaminantes Ambientales/toxicidad , Jatropha/efectos de los fármacos , Jatropha/fisiología , Metales Pesados/toxicidad , Enfermedades de las Plantas/inducido químicamente , Estrés Fisiológico , HidroponíaRESUMEN
Cellular differentiation is associated with dynamic chromatin remodeling in establishing a cell-type-specific epigenomic landscape. Here, we find that mouse testis-specific and replication-dependent histone H3 variant H3t is essential for very early stages of spermatogenesis. H3t gene deficiency leads to azoospermia because of the loss of haploid germ cells. When differentiating spermatogonia emerge in normal spermatogenesis, H3t appears and replaces the canonical H3 proteins. Structural and biochemical analyses reveal that H3t-containing nucleosomes are more flexible than the canonical nucleosomes. Thus, by incorporating H3t into the genome during spermatogonial differentiation, male germ cells are able to enter meiosis and beyond.
Asunto(s)
Histonas/genética , Espermatogonias/metabolismo , Testículo/metabolismo , Proteínas Adaptadoras Transductoras de Señales/genética , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Secuencia de Aminoácidos , Animales , Azoospermia/etiología , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Diferenciación Celular , Células Cultivadas , Cromatina/química , Cromatina/metabolismo , Células Germinativas/citología , Células Germinativas/metabolismo , Haploidia , Histonas/química , Histonas/deficiencia , Masculino , Meiosis , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Nucleosomas/química , Nucleosomas/metabolismo , Espermatogénesis , Testículo/patologíaRESUMEN
The molecular mechanisms of neuronal morphology and synaptic vesicle transport have been largely elusive, and only a few of the molecules involved in these processes have been identified. Here, we developed a novel morphology-based gene trap method, which is theoretically applicable to all cell lines, to easily and rapidly identify the responsible genes. Using this method, we selected several gene-trapped clones of rat pheochromocytoma PC12 cells, which displayed abnormal morphology and distribution of synaptic vesicle-like microvesicles (SLMVs). We identified several genes responsible for the phenotypes and analyzed three genes in more detail. The first gene was BTB/POZ domain-containing protein 9 (Btbd9), which is associated with restless legs syndrome. The second gene was cytokine receptor-like factor 3 (Crlf3), whose involvement in the nervous system remains unknown. The third gene was single-stranded DNA-binding protein 3 (Ssbp3), a gene known to regulate head morphogenesis. These results suggest that Btbd9, Crlf3, and Ssbp3 regulate neuronal morphology and the biogenesis/transport of synaptic vesicles. Because our novel morphology-based gene trap method is generally applicable, this method is promising for uncovering novel genes involved in the function of interest in any cell lines.
Asunto(s)
Regulación de la Expresión Génica/fisiología , Mutagénesis Insercional/métodos , Neuronas/citología , Neuronas/metabolismo , Animales , Toxinas Bacterianas , Southern Blotting , Clonación Molecular , Técnicas de Silenciamiento del Gen , Vectores Genéticos , Cariotipo , Células PC12 , Proteínas Citotóxicas Formadoras de Poros , ARN Interferente Pequeño , Ratas , Retroviridae , Factores de TranscripciónAsunto(s)
Inmunoglobulina G/metabolismo , Linfoma Inmunoblástico de Células Grandes/diagnóstico , Linfoma Inmunoblástico de Células Grandes/metabolismo , Neoplasias de Células Plasmáticas/diagnóstico , Neoplasias de Células Plasmáticas/metabolismo , Anciano de 80 o más Años , Femenino , Fluorodesoxiglucosa F18 , Humanos , Inmunofenotipificación , Imagen Multimodal , Tomografía de Emisión de Positrones , Tomografía Computarizada por Rayos XRESUMEN
Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+)ALL) is one of the highest-risk ALL groups. Whenever possible, patients with Ph(+)ALL should undergo allogeneic hematopoietic stem cell transplantation (HSCT) after induction of remission. Although unrelated cord blood transplantation (CBT) has become a common treatment in adult patients who lack a sibling donor, data on the efficacy of CBT for Ph(+)ALL are limited. We analyzed the clinical outcomes of 20 Ph(+)ALL patients who underwent CBT (n = 8) or unrelated bone marrow transplantation (BMT) (n = 12). The median age was 41 years (range, 17-55 years). All but one of the patients were treated with an imatinib-based regimen before HSCT, and 19 patients were in first complete remission (CR) and 1 patient was in second CR at the time of HSCT. Seventeen patients received a myeloablative conditioning regimen containing 12 Gy of total-body irradiation, and 3 received a reduced-intensity conditioning regimen. After a median of 26 months of follow-up, estimated 3-year overall and leukemia-free survival rates were 100% and 85%, respectively, after CBT, and 49% and 38%, respectively, after unrelated BMT. The CBT group had significantly better overall survival than the BMT group (P = .02). Although BCR-ABL transcript was detected in 4 of 8 CBT patients at transplantation, 7 patients remained in molecular CR. Our findings suggest that CBT may be a viable option as postinduction therapy for Ph(+)ALL in patients lacking a sibling donor.
Asunto(s)
Trasplante de Células Madre de Sangre del Cordón Umbilical , Leucemia-Linfoma Linfoblástico de Células Precursoras/cirugía , Donantes de Tejidos , Trasplante Homólogo/métodos , Adolescente , Adulto , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Biomarcadores de Tumor/sangre , Terapia Combinada , Trasplante de Células Madre de Sangre del Cordón Umbilical/estadística & datos numéricos , Femenino , Proteínas de Fusión bcr-abl/sangre , Enfermedad Injerto contra Huésped/epidemiología , Humanos , Estimación de Kaplan-Meier , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Agonistas Mieloablativos/uso terapéutico , Neoplasia Residual , Leucemia-Linfoma Linfoblástico de Células Precursoras/sangre , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamiento farmacológico , Leucemia-Linfoma Linfoblástico de Células Precursoras/patología , Inducción de Remisión , Estudios Retrospectivos , Acondicionamiento Pretrasplante/métodos , Resultado del Tratamiento , Irradiación Corporal Total , Adulto JovenRESUMEN
The prognosis of advanced extranodal NK/T cell lymphoma (ENKTL) is poor. Allogeneic hematopoietic stem cell transplant (allo-HSCT) has been suggested to be a promising treatment for this disease, but its utility has yet to be established. Here we retrospectively analyzed five cases of ENKTL treated with allo-HSCT in our institute. After induction chemotherapy, disease status at allo-HSCT was second CR in three patients and refractory in two patients. All patients received a myeloablative conditioning regimen, and GVHD prophylaxis consisted of tacrolimus or cyclosporine with short-term methotrexate. Only one patient who received conventional induction chemotherapy developed severe complications, which needed long-term treatment, while others who received chemotherapy containing l-asparaginase did not have severe complications. There were no cases of treatment-related mortality, and all patients survived without disease for a median follow-up period of 1911 days. These results suggested that allo-HSCT following l-asparaginase-containing induction chemotherapy might improve the outcome of advanced ENKTL.
Asunto(s)
Asparaginasa/uso terapéutico , Resistencia a Antineoplásicos , Enfermedad Injerto contra Huésped/prevención & control , Trasplante de Células Madre Hematopoyéticas , Linfoma de Células T/terapia , Células T Asesinas Naturales/patología , Neoplasias Nasales/terapia , Adulto , Antineoplásicos/uso terapéutico , Terapia Combinada , Ciclosporina/uso terapéutico , Femenino , Humanos , Inmunosupresores/uso terapéutico , Linfoma de Células T/patología , Masculino , Metotrexato/uso terapéutico , Recurrencia Local de Neoplasia/patología , Recurrencia Local de Neoplasia/terapia , Neoplasias Nasales/patología , Inducción de Remisión , Estudios Retrospectivos , Terapia Recuperativa , Tasa de Supervivencia , Trasplante Homólogo , Resultado del Tratamiento , Adulto JovenRESUMEN
A case of severe autoimmune hemolytic anemia after ABO matched living donor liver transplantation. We present here a case of severe autoimmune hemolytic anemia after ABO matched living donor liver transplantation. The patient is 56 y.o male. He received living donor liver transplantation from his ABO matched son in May 2007. In July, he was suffered from a progressive anemia, and diagnosed as autoimmune hemolytic anemia by the laboratory examinations. Intensive treatment including predonisolone, azathiopurine, rituximab, plasma exchange, was given, however, the disease was resistant to the treatment. By the administration of cyclophosphamide combined with rituximab, remission was finally achieved. To date, immune mediated hemolytic anemia after ABO matched living donor liver transplantation has not been reported, although several cases of ABO mismatched living donor liver transplantation have been reported. His severe but transient clinical course of anemia suggests that the transient emergence of donor lymphocytes may be responsible for onset of hemolytic anemia.
Asunto(s)
Anemia Hemolítica Autoinmune/etiología , Trasplante de Hígado/efectos adversos , Donadores Vivos , Sistema del Grupo Sanguíneo ABO , Anemia Hemolítica Autoinmune/terapia , Incompatibilidad de Grupos Sanguíneos , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Acquired hemophilia A is a rare and potentially fatal condition of coagulopathy caused by autoantibodies against clotting factor VIII (factor VIII inhibitor). We report a case of a 63-year-old woman, who presented with a sudden onset of severe hemorrhagic tendency with exclusively prolonged activated partial thromboplastin time (APTT). She was diagnosed with acquired hemophilia A due to a decrease in factor VIII activity and a high titer of factor VIII inhibitor. Hemorrhage was well controlled by recombinant activated factor VII. Although the level of factor VIII inhibitor did not decline with prednisolone and cyclophosphamide, it became undetectable with rituximab. In parallel with controlling hemorrhage, malignancy, which may cause acquired hemophilia A, was searched for and sigmoid colon cancer was found. After the eradication of factor VIII inhibitor, surgical resection was performed uneventfully. Thereafter, acquired hemophilia A has been in complete remission without any additional therapy. The present case suggests the efficacy of rituximab for refractory acquired hemophilia A and the importance of the identification of underlying diseases that can cause acquired hemophilia A.
Asunto(s)
Anticuerpos Monoclonales/administración & dosificación , Antineoplásicos/administración & dosificación , Hemofilia A/tratamiento farmacológico , Hemofilia A/etiología , Neoplasias del Colon Sigmoide/complicaciones , Neoplasias del Colon Sigmoide/terapia , Anticuerpos Monoclonales de Origen Murino , Inhibidores de Factor de Coagulación Sanguínea/sangre , Colon Sigmoide/cirugía , Factor VIII/análisis , Femenino , Hemofilia A/sangre , Humanos , Persona de Mediana Edad , Tiempo de Tromboplastina Parcial , Rituximab , Neoplasias del Colon Sigmoide/sangreRESUMEN
Bortezomib is approved for the treatment of patients with relapsed or refractory multiple myeloma (MM), but only a few clinical studies for Japanese patients who were treated with bortezomib have been reported. We retrospectively analyzed 40 patients with relapsed or refractory MM who have received bortezomib at three collaborating centers in Miyagi prefecture in Japan. All the patients have been received bortezomib in combination with dexamethasone. Responses were determined using International Myeloma Working Group uniform response criteria. The overall response was observed in 30 patients (75%), including very good partial response in 8 patients (20%), and partial response in 22 patients (55%). The median time to disease progression was 8.7 months, and the median overall survival has not been reached. The factors affecting time to disease progression were International Staging System stage, serum beta2-microglobulin level, and number of treatment cycles. The most common grade 3 and 4 adverse events were thrombocytopenia (50%), peripheral neuropathy (25%), leukopenia (25%), and herpes zoster infection (25%). Thus, bortezomib is well tolerated and effective for Japanese patients with relapsed or refractory MM. Our results suggest that serum beta2-microglobulin level may be a marker of prognosis on bortezomib therapy for patients with relapsed or refractory MM although further studies are needed.
Asunto(s)
Antineoplásicos/uso terapéutico , Ácidos Borónicos/uso terapéutico , Mieloma Múltiple/tratamiento farmacológico , Mieloma Múltiple/metabolismo , Pirazinas/uso terapéutico , Microglobulina beta-2/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Antineoplásicos/efectos adversos , Antineoplásicos/farmacología , Ácidos Borónicos/efectos adversos , Ácidos Borónicos/farmacología , Bortezomib , Progresión de la Enfermedad , Femenino , Humanos , Japón , Masculino , Persona de Mediana Edad , Mieloma Múltiple/patología , Pirazinas/efectos adversos , Pirazinas/farmacología , Recurrencia , Estudios Retrospectivos , Factores de Tiempo , Resultado del TratamientoRESUMEN
Real-time quantitative polymerase chain reaction (RQ-PCR) has been accepted as integral part of the management of patients with hematologic malignancies. Whereas standardization efforts of RQ-PCR, initiated by Europe Against Cancer (EAC) group, have been gradually widespread in the world, Japanese laboratories use their individual protocol for RQ-PCR analysis. Therefore, we assessed the variability of quantitative results obtained from 4 different laboratories in Japan, including 3 companies and Tohoku University Hospital, using identical peripheral blood or bone marrow samples of patients in chronic myeloid leukemia (CML; n = 11) and acute myeloid leukemia (AML; n = 2). RQ-PCR was designed to quantify the copy numbers of disease-specific fusion chimeras; BCR-ABL (CML) and AML1-ETO (AML). In 5 out of 13 samples, the quantitative results from 4 laboratories varied more than 10 times (up to 712 times). Thus, we next sought to determine factors affecting the variability of RQ-PCR results across laboratories, by sending back RNA and cDNA samples from each company to Tohoku University, and they were further proceed to yield quantitative data. The main difference between companies and Tohoku University was probably due to the difference of blood separation method (Blood lysis or Ficoll-Hypaque). On the other hand, the variability among 4 laboratories was the most noticeable in the PCR step, mainly attributable to the difference of primer/probe sequence among laboratories. In conclusion, our analyses indicate the importance to limit both preanalytical (sample processing) and analytical (RQ-PCR) interlaboratory variability for RQ-PCR protocol, and the need of further efforts on standardization program in Japan.
Asunto(s)
Leucemia/diagnóstico , Leucemia/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/normas , Humanos , Japón , Neoplasia Residual/diagnóstico , Neoplasia Residual/genética , Estándares de Referencia , Reproducibilidad de los ResultadosRESUMEN
Dual surface immunoglobulin light-chain expression in B-cell malignant neoplasm is a rare event, and has been predominantly reported in chronic lymphocytic leukemia. Herein, we report a case of aggressive B-cell lymphoma with kappa/lambda-dual surface immunoglobulin light-chain expression of a 69-year-old woman. The lymphoma cells were positive for CD5, CD19, CD20, HLA-DR, Ig kappa and Ig lambda. Southern blot analysis confirmed rearranged bands for both light chains with a monoclonal heavy chain rearrangement. She was treated with a combination of rituximab and CHOP regimen, but died of the progressive disease. To our knowledge, this is the first case of aggressive B-cell lymphoma showing dual kappa/lambda expression; the possible mechanisms of abnormal light chain expression are discussed.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Cadenas Ligeras de Inmunoglobulina/biosíntesis , Linfoma de Células B/tratamiento farmacológico , Linfoma de Células B/inmunología , Anciano , Anticuerpos Monoclonales/administración & dosificación , Anticuerpos Monoclonales de Origen Murino , Ciclofosfamida/administración & dosificación , Progresión de la Enfermedad , Doxorrubicina/administración & dosificación , Resultado Fatal , Femenino , Humanos , Linfoma de Células B/fisiopatología , Prednisolona/administración & dosificación , Rituximab , Vincristina/administración & dosificaciónRESUMEN
Nasal natural killer (NK)/T cell lymphoma is a rare entity of non-Hodgkin's lymphoma which mostly occurs in East Asian countries. The advanced disease above clinical stage III is often refractory to the radiation and chemotherapies, remission is transient even if achieved, and median survival is about 12 months. Thus the prognosis of advanced NK/T cell lymphoma is generally poor, however, the promising results of allogeneic hematopoietic stem cell transplantation for advanced NK/T cell lymphoma have been recently reported. In most of these cases, stem cell sources were human leukocyte antigen (HLA) matched donors and alternative sources were seldom used. We report here a case of a 36-year-old woman who was diagnosed as having an extranodal NK/T cell lymphoma, nasal type. The patient achieved a complete remission after 2 cycles of chemotherapy including Carboplatin, Etoposide, Ifosfamide, and Dexamethasone, but 3-months later relapsed during the search for HLA-matched unrelated donors. She received unrelated cord blood transplantation (CBT) in the second remission achieved by a regimen containing L-asparaginase. The conditioning regimen was 12 Gy of total body irradiation, high-dose cytarabin and cyclophosphamide. FK506 and methotrexate were used for graft-versus-host disease (GVHD) prophylaxis. GVHD involving the intestine and the oral mucosa was observed, but improved without additional immunosuppressive therapies. The patient remains in remission 33 months after CBT. Cord blood thus could be an appropriate stem cell source for patients with advanced NK/T lymphoma who have no HLA matched donors.
Asunto(s)
Trasplante de Células Madre de Sangre del Cordón Umbilical , Linfoma de Células T/terapia , Adulto , Femenino , Prueba de Histocompatibilidad , Humanos , Células Asesinas Naturales/patología , Linfoma de Células T/patología , Neoplasias Nasales/patología , Neoplasias Nasales/terapia , Donantes de TejidosRESUMEN
Chronic myelogenous leukemia (CML) is a hematological malignancy that is characterized by the chromosome anomaly, t(9;22)(q34;q11). By this chromosomal translocation, a novel activated tyrosine kinase, BCR-ABL chimeric protein, is generated, and the protein is causative of the disease. Recently, Imatinib mesylate targeting to a BCR-ABL chimeric protein has been developed, and shown to achieve complete remission at a high rate. Patients are currently required to receive a fixed dose, 400 mg daily; however, it is possible that some of patients can maintain their remission with reduced doses of imatinib. In this study, we determined levels of BCR-ABL transcript in CML patients by real-time quantitative polymerase chain reaction analysis, and explored the possibility of individualization of therapeutic doses of imatinib. Thirty-five CML patients, including 17 newly diagnosed patients, 16 patients pre-treated with interferon-alpha, and 2 relapsed patients after allogeneic transplantation, were treated with imatinib. Complete cytogenetic response was achieved in 31 (89%) patients. Major molecular response (MMR) was achieved in 21 (60%). Complete molecular response (CMR) was achieved in 7 (20%). Imatinib was discontinued in 2 patients, one patient with MMR due to noncompliance and other patient sustaining CMR, but both patients relapsed 7 and 13 months later, respectively. The doses of imatinib were reduced in 7 patients due to its side effects, but 4 out of the 7 patients have sustained MMR, and 2 of them have sustained CMR for more than 23 months. These results indicate that some patients are able to maintain MMR with low-dose imatinib.
Asunto(s)
Antineoplásicos/administración & dosificación , Proteínas de Fusión bcr-abl/sangre , Leucemia Mielógena Crónica BCR-ABL Positiva/tratamiento farmacológico , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Recurrencia Local de Neoplasia/tratamiento farmacológico , Piperazinas/administración & dosificación , Pirimidinas/administración & dosificación , Adulto , Anciano , Benzamidas , Biomarcadores de Tumor/análisis , Relación Dosis-Respuesta a Droga , Esquema de Medicación , Femenino , Humanos , Mesilato de Imatinib , Leucemia Mielógena Crónica BCR-ABL Positiva/sangre , Masculino , Persona de Mediana Edad , Planificación de Atención al Paciente , Reacción en Cadena de la Polimerasa , Inducción de Remisión , Resultado del TratamientoRESUMEN
Nucleolar spindle-associated protein (NuSAP), a recently characterised microtubule-associated protein, appears to participate in cell cycle regulation. It has been demonstrated that NuSAP is expressed preferentially in the erythroid lineage in haematopoietic cells. To characterise its role in erythropoiesis, we examined the expression profile of the NuSAP gene. In fractionated murine erythroblasts, NuSAP mRNA was remarkably more abundant in the subset corresponding to immature erythroblasts (TER119(+)CD71(high)) than mature erythroblasts (TER119(+)CD71(low)), and it was significantly increased in TER119(+) cells from in vivo phlebotomised mice compared with control mice. Furthermore, during erythroid maturation of mouse erythroleukaemia (MEL) cells by dimethylsulfoxide, NuSAP mRNA was increased at 24-72 h. These results suggested that the NuSAP gene might contribute to the expansion of immature erythroblast pool. The regulatory mechanism of NuSAP gene was investigated using MEL cells. Sequence analysis revealed that NuSAP promoter has four CCAAT boxes, an Sp1 element, a GATA-like element, a CACCC element, a Myb element and lacks a TATA box. Promoter analyses demonstrated that duplicated CCAAT boxes located at -81/-85 and -30/-34 were essential for promoter activity. Furthermore, the promoter was trans-activated by NF-YA through these elements. These results suggest that NuSAP might play an important role in erythroid proliferation under the control of NF-Y.