RESUMEN
BACKGROUND: Serum and urinary protein electrophoresis play an important role in the identification of monoclonal gammopathy. Recently, capillary electrophoresis (CE) has been adapted in many clinical laboratories because of several advantages such as short turnaround time, automation, and high reproducibility. However, there have been unsolved concerns for the concordance between conventional gel and automated capillary electrophoresis methods for protein separation in clinical specimens. In this study, we investigated the diagnostic performance of both methods for detecting monoclonal (M) protein. METHODS: From February 2012 to August 2015, a total of 3,013 CE tests were performed in our hospital. Among these cases, we reconfirmed results of CE (Capillary 2, Sebia, Lysse, France) with those of conventional agarose gel electrophoresis (GE) (Hydragel 4IF, Sebia, Lisses, France) in 28 specimens from 24 patients with newly diagnosed monoclonal gammopathy (group 1). In addition, 22 cases from 15 patients with previously diagnosed monoclonal gammopathy presenting indeterminate or suspicious results on CE (group 2) were also reconfirmed with GE. RESULTS: We compared the results between the two electrophoresis methods in two different groups of patients with newly diagnosed discrete monoclonal peaks vs. pre-existing monoclonal gammopathy with obscure results in follow-up courses. In group 1, agreement rate was 100% (28/28) and there was no discrepant result between these two electrophoresis methods. In contrast, group 2 showed 86.4% (19/22) agreement rate and 0.67 Cohen's kappa value (95% confidence interval, 0.51 - 1.02). CONCLUSIONS: According to our results, both electrophoresis methods can be used with the same level of assurance at the time of initial diagnosis for monoclonal gammopathy. However, in patients with previously diagnosed monoclonal gammopathy in follow-up course after appropriate treatments, discordant results can be observed due to the reduced amount of M proteins. Therefore, we suggest that some ambiguous cases with very small amounts of M components require a combination of both CE and GE methods for accurate interpretation to confirm the presence of M proteins.
Asunto(s)
Servicios de Laboratorio Clínico/normas , Electroforesis en Gel de Agar/métodos , Electroforesis Capilar/métodos , Paraproteinemias/diagnóstico , Electroforesis en Gel de Agar/estadística & datos numéricos , Electroforesis Capilar/estadística & datos numéricos , Humanos , Proteínas de Mieloma/análisis , Paraproteinemias/sangre , Paraproteinemias/orina , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
To investigate the sequence types (STs) and fluoroquinolone resistance related mutations among ciprofloxacin (CIP)-non-susceptible extended-spectrum ß-lactamase (ESBL)-producing E. coli isolated from Korean patients from 2006-2008. The prevalence of fluoroquinolone resistance-determining region (QRDR) mutations in gyrA, gyrB, parC, and parE and plasmid-mediated quinolone resistance (PMQR) genes were also studied. Multilocus sequence typing (MLST) was performed to identify STs. The most common ST was ST131 (33/51, 64.7%). All isolates, except one isolate, showed three mutations at codons 83 (S83L) and 87 (S87N) in gyrA and 80 (S80I) in parC The prevalence of ST131 in our hospital was much higher than reported in other Asian studies during a similar time period. The mutations found in ST131 were concordant with other studies.
Asunto(s)
Ciprofloxacina/farmacología , Farmacorresistencia Bacteriana/genética , Infecciones por Escherichia coli/epidemiología , Escherichia coli/aislamiento & purificación , Fluoroquinolonas/farmacología , Genes Bacterianos , Tipificación de Secuencias Multilocus/métodos , Antibacterianos/farmacología , Estudios Epidemiológicos , Escherichia coli/genética , Infecciones por Escherichia coli/genética , Infecciones por Escherichia coli/microbiología , Hospitales Universitarios , Humanos , Pruebas de Sensibilidad Microbiana , Mutación , Pronóstico , República de Corea/epidemiología , Atención Terciaria de Salud , beta-Lactamasas/metabolismoAsunto(s)
Sistema del Grupo Sanguíneo ABO/genética , Tipificación y Pruebas Cruzadas Sanguíneas , Quimerismo , Electroforesis en Gel de Poliacrilamida , Adulto , Femenino , Genotipo , Heterocigoto , Humanos , Repeticiones de Microsatélite , Fenotipo , Análisis de Secuencia de ADN , Eliminación de SecuenciaRESUMEN
The inhibin-α gene was identified as a tumor suppressor gene in the gonads and adrenal glands by functional studies using knockout mice. Methylation of CpG sites within the regulatory regions of tumor suppressor gene is frequently associated with their transcriptional silencing. We investigated epigenetic modifications, changes in loss of heterozygosity (LOH), and mutation of the inhibin-α gene, and regulation of transcriptional expression in response to inhibitors of DNA methylation (5-aza-2'-deoxycytidine, 5-AzaC) in human lymphoid (Jurkat, Molt-4, Raji, and IM-9) and myeloid (HL-60, Kasumi-1, and K562) leukemia cells. The inhibin-α promoter was hypermethylated in lymphoid (Molt-4 and Raji) and myeloid (HL-60 and Kasumi-1) leukemia cells. Inhibin-α gene mutations differed significantly between lymphoid (heterozygote) and myeloid (homozygote) leukemia cells. LOH in the inhibin-α gene was detected in lymphoid and myeloid leukemia cells, with the exception of Jurkat cells. Treatment with 5-AzaC, a demethylating agent, resulted in increased inhibin-α mRNA and protein levels in most of the cell lines. Also, 5-AzaC treatment inhibited cell proliferation and induced apoptosis. Taken together, our results reveal that the inhibin-α gene is transcriptionally silenced in human leukemia cells and that reactivation is suppressed by a demethylating agent. In addition, mutations in, and expression levels of, the inhibin-α gene differed between human lymphoid and myeloid leukemia cells.
Asunto(s)
Metilación de ADN , Epigénesis Genética , Regulación Neoplásica de la Expresión Génica , Inhibinas/genética , Leucemia Linfoide/genética , Leucemia Mieloide/genética , Apoptosis/efectos de los fármacos , Azacitidina/análogos & derivados , Azacitidina/farmacología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Islas de CpG/genética , Metilación de ADN/efectos de los fármacos , Decitabina , Inhibidores Enzimáticos/farmacología , Silenciador del Gen , Humanos , Pérdida de Heterocigocidad , Mutación , Regiones Promotoras Genéticas , ARN Mensajero/metabolismoRESUMEN
BACKGROUND: We investigated mutations in the quinolone resistance-determining region (QRDR) of ciprofloxacinnonsusceptible extended-spectrum ï¢-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae by a statistical analysis. METHODS: We collected 97 clinical isolates of ciprofloxacin-nonsusceptible ESBL-producing E. coli (55 strains) and K. pneumoniae (42 strains) from a tertiary-care university hospital in Seoul, Republic of Korea, between 2006 and 2008. The QRDR of the gyrA, gyrB, parC, and parE genes were amplified by PCR and sequenced. RESULTS: Most E. coli isolates (53/55; 96.4%) with a minimum inhibitory concentration of ≥ 64 mg/L against ciprofloxacin had double mutations in gyrA (Ser-83ï®Leu and Asp-87ï®Asn) and at least one mutation in parC (Ser-80 ï®Ile or Glu-84ï®Val), with or without one in parE. Fifty E. coli (90.9%) isolates had a mutation in parE, of which Ile-529ï®Leu (70.9%) was the most frequent. However, we could not find statistically significant variables in increasing ciprofloxacin resistance in E. coli isolates. Thirty-six K. pneumoniae isolates (36/42; 85.7%) had at least one mutation in gyrA, gyrB, or parC, and the mutation in gyrA might have been associated with plasmid-mediated quinolone-resistance (PMQR). Ser-80ï®Ile in parC and aac(6')-Ib-cr in the K. pneumoniae isolates were significantly associated with an increased MIC of ciprofloxacin by ordinal logistic regression analysis. CONCLUSIONS: The Ser-80ï®Ile in parC and aac(6')-Ib-cr in K. pneumoniae are supposed to play an important role in increased ciprofloxacin resistance, but statistically significant variables could not be found in E. coli isolates in the present study.
Asunto(s)
Escherichia coli , Klebsiella pneumoniae , Mutación , Antibacterianos , Ciprofloxacina , Girasa de ADN , Topoisomerasa de ADN IV , Farmacorresistencia Bacteriana , Humanos , Pruebas de Sensibilidad Microbiana , República de CoreaAsunto(s)
Linfoma Folicular , Electroforesis Capilar , Humanos , Inmunoglobulina M , ParaproteinemiasAsunto(s)
Cirrosis Hepática Alcohólica , Asia , Electroforesis Capilar , Humanos , Isoformas de Proteínas , TransferrinaAsunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Cromosomas Humanos Par 12 , Cromosomas Humanos Par 3 , Fusión Génica , Reordenamiento Génico , Leucemia Mieloide Aguda/genética , Proteína del Locus del Complejo MDS1 y EV11/genética , Proteínas Proto-Oncogénicas c-ets/genética , Proteínas Represoras/genética , Translocación Genética , Deleción Cromosómica , Cromosomas Humanos Par 7/genética , Predisposición Genética a la Enfermedad , Humanos , Cariotipificación , Leucemia Mieloide Aguda/inducido químicamente , Leucemia Mieloide Aguda/diagnóstico , Leucemia Mieloide Aguda/tratamiento farmacológico , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa Multiplex , Fenotipo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteína ETS de Variante de Translocación 6Asunto(s)
Síndromes Mielodisplásicos/diagnóstico , Complicaciones Hematológicas del Embarazo/diagnóstico , Complicaciones Neoplásicas del Embarazo/diagnóstico , Adulto , Femenino , Humanos , Síndromes Mielodisplásicos/sangre , Síndromes Mielodisplásicos/genética , Regresión Neoplásica Espontánea , Parto , Valor Predictivo de las Pruebas , Embarazo , Complicaciones Hematológicas del Embarazo/sangre , Complicaciones Hematológicas del Embarazo/genética , Complicaciones Neoplásicas del Embarazo/sangre , Complicaciones Neoplásicas del Embarazo/genética , Recurrencia , Factores de RiesgoAsunto(s)
Infecciones por Bacteroides/diagnóstico , Bacteroides/genética , Chaperonina 60/genética , Girasa de ADN/genética , ADN Bacteriano/genética , ARN Ribosómico 16S/genética , Ribotipificación/métodos , Análisis de Secuencia de ADN , Antibacterianos/uso terapéutico , Bacteroides/clasificación , Bacteroides/efectos de los fármacos , Infecciones por Bacteroides/sangre , Infecciones por Bacteroides/tratamiento farmacológico , Infecciones por Bacteroides/microbiología , Infecciones por Bacteroides/transmisión , Femenino , Humanos , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Resultado del TratamientoAsunto(s)
Aerococcaceae/genética , Bacteriemia/microbiología , Infecciones por Bacterias Grampositivas/microbiología , ARN Ribosómico 16S/genética , Aerococcaceae/aislamiento & purificación , Anciano de 80 o más Años , ADN Bacteriano/análisis , ADN Bacteriano/aislamiento & purificación , Femenino , Humanos , República de Corea , Análisis de Secuencia de ADN/métodosAsunto(s)
Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Vancomicina , Antibacterianos/uso terapéutico , Humanos , Linezolid , Meticilina , Resistencia a la Meticilina , Oxazolidinonas/uso terapéutico , Neumonía Estafilocócica/tratamiento farmacológico , Staphylococcus aureus/efectos de los fármacosAsunto(s)
Albúminas/metabolismo , Albuminuria/diagnóstico , Trastornos de las Proteínas Sanguíneas/diagnóstico , Electroforesis en Gel de Agar/métodos , Electroforesis Capilar/métodos , Albúmina Sérica/metabolismo , Albuminuria/metabolismo , Trastornos de las Proteínas Sanguíneas/metabolismo , HumanosAsunto(s)
Bacteriemia/diagnóstico , Proteínas Bacterianas/genética , Corynebacterium/genética , ARN Ribosómico 16S/genética , Antibacterianos/uso terapéutico , Bacteriemia/tratamiento farmacológico , Bacteriemia/microbiología , Proteínas Bacterianas/metabolismo , Proteína C-Reactiva/análisis , Catéteres Venosos Centrales , Corynebacterium/aislamiento & purificación , Farmacorresistencia Bacteriana Múltiple , Humanos , Masculino , Persona de Mediana Edad , ARN Ribosómico 16S/metabolismoAsunto(s)
Transfusión de Eritrocitos , Sistema del Grupo Sanguíneo Rh-Hr/genética , Globulina Inmune rho(D)/sangre , Adenocarcinoma/sangre , Adenocarcinoma/cirugía , Pueblo Asiatico/genética , Donantes de Sangre , Eliminación de Gen , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Fenotipo , Neoplasias de la Próstata/sangre , Neoplasias de la Próstata/cirugía , Población BlancaRESUMEN
AIM: Although the etiology of plasma cell dyscrasia is poorly understood, there is evidence for immune dysregulation or sustained immune stimulation playing a pivotal role in the pathogenesis of these diseases, including chronic infection and autoimmune disorders. In this study, we report four autoimmune disease cases where monoclonal gammopathy (MG) was incidentally found during follow-up. METHODS: We retrospectively reviewed the medical charts and laboratory test results in the following four cases: neuromyelitis optica, Kikuchi disease, Sjögren syndrome and ankylosing spondylosis. RESULTS: The four patients were older than 55 years and the male-to-female ratio was 2 : 2. The autoimmune disease in each case developed differently because two patients had coincidental detection of MG, whereas MG was detected 2 years and 10 years after diagnosis in the other two patients. The amount of M-components in the blood for two cases was ≤ 1 g/dL. For the other two subjects, M-components were ≥ 3 g/dL. CONCLUSION: A high prevalence of MG of undetermined significance (MGUS) has been noted in a series of patients with immune disorders, suggesting a possible association with MG. Further studies should focus on determining how MG relates to various clinical information and laboratory parameters, such as disease duration, disease activity and higher sedimentation rate. In the future, we also need to identify which stimuli, such as cytokine types and levels, can induce lymphocyte clonal transformation and the production of monoclonal antibodies.