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1.
Med. clín (Ed. impr.) ; 154(7): 275-278, abr. 2020. graf
Artigo em Espanhol | IBECS | ID: ibc-190912

RESUMO

INTRODUCCIÓN Y OBJETIVOS: La analbuminemia congénita (AAC) (MIM #616000) es una enfermedad autosómica recesiva (prevalencia <1/106) causada por defectos en el gen ALB que implican la ausencia o marcada disminución de la albuminemia. En este artículo, describimos un caso de AAC detectado en nuestro hospital. MATERIAL Y MÉTODOS: Mujer de 42 años con hipoproteinemia e hipoalbuminemia de causa no filiada. El estudio bioquímico se realizó siguiendo las técnicas y los controles de calidad habituales de nuestro laboratorio: albuminemia (colorimetría y nefelometría); electroforesis de proteínas (capilar y gel de agarosa) y análisis molecular del gen ALB (extracción de ADN y amplificación PCR de los 14 exones codificantes más regiones intrónicas adyacentes y secuenciación Sanger). RESULTADOS: Descartadas las causas más frecuentes de hipoalbuminemia, se confirmó la analbuminemia por electroforesis y nefelometría. El estudio molecular del gen ALB evidenció la presencia de la variante c.1289+1G>A (variante Guimarães) en homozigosis. CONCLUSIONES: Este es el primer caso confirmado mediante estudio molecular de AAC en España. La paciente presenta la variante Guimarães descrita previamente en otros 4 pacientes en el mundo


INTRODUCTION AND OBJECTIVES: Congenital analbuminaemia (CCA) (MIM #616000) is an autosomal recessive disorder (prevalence < 1/106) caused by defects in the ALB gene leading to absence or severe reduction of albuminaemia. This paper describes a case of CCA detected and diagnosed in our hospital. MATERIALS AND METHODS: A 42-year old woman showing hypoproteinaemia and hypoalbuminaemia of unknown aetiology. Biochemical study was performed according to routine quality controlled analytical procedures: Albuminaemia (colorimetric and nephelometric methods). Protein electrophoresis (capillary and agarose gel). Molecular study of the ALB gene: DNA extraction, PCR amplification of the 14 coding exons plus adjacent intron regions and Sanger sequencing. RESULTS: After discarding the most common causes of hypoalbuminaemia, the analbuminaemia was confirmed by nephelometry and protein electrophoresis. The proband was found to be homozygous for molecular defect in the ALB gene: variant c.1289+1G>A previously reported as Guimarães variant. CONCLUSIONS: This is the first case of CCA confirmed by molecular study in Spain. The proband shows the Guimarães variant previously described in 4 patients worldwide


Assuntos
Humanos , Feminino , Adulto , Doenças Genéticas Inatas/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Albumina Sérica/genética , Doenças Genéticas Inatas/genética , Eletroforese , Hipoalbuminemia/etiologia , Colorimetria/métodos , Nefelometria e Turbidimetria/métodos , Diagnóstico Diferencial
2.
Allergol. immunopatol ; 48(1): 26-33, ene.-feb. 2020. ilus, graf
Artigo em Inglês | IBECS | ID: ibc-186588

RESUMO

Introduction and Objectives: The production and consumption of oysters is increasing annually because it can provide essential nutrients and benefit for human health, leading to frequent occurrence of severe allergic reactions observed in sensitized individuals. The aim of the present study was to investigate the effects of acid and protease treatment on the conformation and IgE-binding capacity of recombinant Crassostrea gigas tropomyosin (Cra g 1). Results: Under acidic conditions, Cra g 1 did not undergo degradation, however, the changes obvious in the intensity of CD signal and ANS-binding fluorescence were observed, which was associated with a decrease in antibody reactivity. In simulated gastrointestinal fluid (SGF) and simulated intestinal fluid (SIF) digestion system, acid-treated Cra g 1 was relatively resistant to digestion, but the degradative patterns were very different. Moreover, owing to alterations of secondary structure and hydrophobic surface of the protein during digestive processing, antigenicity of acid-induced Cra g 1 reduced in SGF while it increased significantly in SIF. Conclusion: To our knowledge, this is the first study reporting that antigenicity of acid-treated oyster tropomyosin increased after SIF digestion. These results revealed that treatment with acid and pepsin, rather than trypsin, was an effective way of reducing IgE-binding capacity of tropomyosin from oyster


No disponible


Assuntos
Humanos , Imunoglobulina E/imunologia , Técnicas In Vitro/métodos , Alérgenos/química , Tropomiosina/química , Hipersensibilidade Alimentar/imunologia , Digestão , Alérgenos/imunologia , Tropomiosina/imunologia , Ostreidae/imunologia , Sistema Digestório/imunologia , Eletroforese/métodos , Análise Espectral/métodos , Ensaio de Imunoadsorção Enzimática
4.
Ars pharm ; 60(4): 219-225, oct.-dic. 2019. graf
Artigo em Espanhol | IBECS | ID: ibc-188485

RESUMO

Introducción: Uno de los fármacos de primera línea en el tratamiento de la esquistosomiasis es el Praziquantel. Numerosas son las formas farmacéuticas sólidas orales desarrolladas hasta la fecha, siendo éstas poco adecuadas para determinados grupos de población, ej. tercera edad y Pediatría, y Veterinaria. Este trabajo describe los primeros pasos en el desarrollo de un estudio de preformulación dirigido al diseño de una forma farmacéutica líquida de administración oral para este principio activo. Método: Se caracterizó la forma y tamaño de las partículas de Praziquantel con las que se pretendía preparar una suspensión acuosa, mediante microscopía electrónica de barrido. Además, se analizó el efecto que el pH y el tipo de electrolito y su concentración tenían sobre el comportamiento de las suspensiones formuladas, gracias a medidas de electroforesis (potencial zeta) y espectrofotometría ultravioleta-visible (turbidimetría en función del tiempo). Resultados: La población de partículas de fármaco se caracterizó por una forma acicular y un tamaño micrométrico, con una distribución de tamaños heterogénea. Se comprobó cómo controlando la composición del medio de dispersión, en términos de pH y electrolitos, podía definirse la carga eléctrica superficial de las partículas de fármaco y, así su proceso de sedimentación, obteniéndose el sistema más adecuado para la vía de administración oral (sistema floculado). Conclusiones: Se han definido las condiciones iniciales de formulación de suspensiones acuosas de Praziquantel destinadas a la vía oral. Un control adecuado de la composición de la fase externa resulta fundamental en el establecimiento del mejor sistema (floculado) para esta vía de administración


Introduction: One of the first-line drugs against schistosomiasis is Praziquantel. Up to now, numerous oral solid dosage forms have been developed, being they considered of little help to elder patients, pediatrics, and Veterinary. Initial steps in the development of preformulation studies aiming the design of a Praziquantel liquid pharmaceutical dosage form to be administered orally are described. Method: Size and shape of Praziquantel particles were characterized by scanning electron microscopy. Furthermore, it was investigated the effect of pH and type of electrolyte and its concentration in the aqueous dispersion media on the behaviour of the suspensions. To that aim, electrokinetic determinations (zeta potential) and ultraviolet-visible spectrophotometry measurements (turbidimetry as a function of time) were done. Results: Drug particles were characterized by an acicular shape and a micrometer size (heterogeneous size distribution). It was observed that controlling the composition of the aqueous dispersion media, in terms of pH and electrolytes, helped in defining the surface electrical charge of the drug particles and, thus the sedimentation profile, obtaining the more adequate system for the oral route of drug administration (flocculated system). Conclusions: Initial conditions to formulate aqueous Praziquantel suspensions for the oral route have been defined. An appropriate control of the composition of the external phase of the suspension is a key aspect when establishing the best liquid pharmaceutical system (flocculated) for this administration route


Assuntos
Humanos , Composição de Medicamentos/métodos , Praziquantel/farmacologia , Suspensão Aquosa/métodos , Estabilidade de Medicamentos , Praziquantel/síntese química , Praziquantel/uso terapêutico , Eletroforese/métodos , Sedimentação , Sedimentação/análise , Eletrólitos/farmacologia
5.
Eur. j. anat ; 23(6): 393-403, nov. 2019. graf, ilus, tab
Artigo em Inglês | IBECS | ID: ibc-185082

RESUMO

Hepatitis C is a widely distributed problem all over the world, especially Egypt. Chronically infected people develop serious liver disease and now it is the most common cause for liver transplantation. Recently, a new regimen, sofosbuvir (sovaldi), alone or with combinations as sovaldi-ribavirin, was approved for treating this disease. There are limited studies that explore the effects of these drugs on the reproductive organs, and hence affection of male fertility while using these drugs. This study aims to throw more light on whether sovaldi or sovaldi-ribavirin causes testicular damaging effects in the adult male albino rats. We investigated the effect of this regimen in a dose equivalent to that used in the human (41 mg/kg once daily orally for sovaldi and 41 mg/kg twice daily orally for ribavirin) for consecutive 5 and 10 days. There was highly significant decrease in testosterone hormone level and marked degenerative changes in the seminiferous tubules and the testicular interstitium, with increase in collagen deposits in sovaldi treated rats, and in a more extensive manner in sovaldi-ribavirin treated rats. There was a significant increase of deoxyribonucleic acid (DNA) fragmentation in the treated groups after 10 days. However, there was a non-significant difference in DNA fragmentation in the treated groups after 5 days when compared with control. Immuno-histochemistry detection of caspase-3 showed significant increase in its expression in the treated groups after either 5 or 10 days. This denoted the specificity of caspase-3 immunohistochemistry technique in the detection of early apoptotic changes. It was concluded that sovaldi and sovaldi ribavirin induced gonado toxic effects through induction of DNA fragmentation via up regulation of caspase-3, and that the resulting damaging effects increased with longer duration of drug in take


No disponible


Assuntos
Animais , Ratos , Sofosbuvir/administração & dosagem , Hepatite C/induzido quimicamente , Hepatite C/veterinária , Testículo/efeitos dos fármacos , Ribavirina/administração & dosagem , Disruptores Endócrinos/administração & dosagem , Sofosbuvir/toxicidade , Imuno-Histoquímica , Projetos de Pesquisa , Eletroforese/métodos
6.
Rev. esp. patol. torac ; 31(2): 144-152, jun. 2019. ilus, graf
Artigo em Espanhol | IBECS | ID: ibc-183656

RESUMO

El cáncer de pulmón (CP) es la primera causa de muerte por cáncer en el mundo. Su elevada mortalidad refleja, en parte, la limitada eficacia de las terapias actualmente disponibles. Dada la pobre supervivencia actual del CP, y la demostrada evidencia de que el diagnóstico precoz reduce la mortalidad, ha cobrado especial interés la búsqueda de biomarcadores. Recientemente, se ha atribuido a DYRK2 un papel relevante en el desarrollo y progresión tumoral relacionado con la inducción de la apoptosis en respuesta al estrés oncogénico. Así, se identificó a DYRK2 como el gen más frecuentemente sobre- expresado en el adenocarcinoma de pulmón, siendo además un factor pronóstico favorable en estos tumores. Asimismo, se ha demostrado la existencia de una regulación mutua entre DYRK2 y la ubiquitín-ligasa SIAH2, en el control de la respuesta a hipoxia y el daño genotóxico. La búsqueda de nuevas dianas y estrategias terapéuticas supone un paso clave para la lucha contra el cáncer de pulmón. El objetivo del trabajo fue investigar qué papel juegan las proteínas SIAH2-DYRK2 en la carcinogénesis pulmonar, así como determinar el impacto clínico-patológico de su expresión en el tejido neoplásico. La modulación de la ruta SIAH2-DYRK2 podría ser empleada como una terapia dirigida en pacientes con cáncer de pulmón


Lung cancer (LC) continues to be the leading cause of cancer-related mortality worldwide. The high mortality highlights the limited efficacy of available therapies for LC treatment. Given the poor survival rate of LC, and considering that early diagnosis reduces mortality, special interest exists nowadays in searching for lung cancer biomarkers. Recently, it has been suggested a possible role of DYRK2 in the development and progression of tumors, related to the induction of apoptosis in response oncogenic stress. In this regard, DYRK2 was identified as the most commonly up-regulated gene in lung adenocarcinomas, as well as a favourable prognostic factor in these tumors. Moreover, a mutual regulation between DYRK2 and the ubiquitin-ligase SIAH2 in response to hypoxia and DNA-damage signaling pathways has been demonstrated. It is of paramount importance to search for new targets and therapeutic strategies in lung cancer. The aim of the study was to analyse the role of SIAH2-DYRK2 in the development of lung cancer, and to assess the clinical and pathological effects of the expression of these proteins in lung cancer tissue. Modulation of the route SIAH2- DYRK2 might be used as a new targeted therapy in lung cancer patients


Assuntos
Humanos , Carcinogênese , Neoplasias Pulmonares/diagnóstico , Prognóstico , Diagnóstico Precoce , Carcinoma de Células Escamosas/diagnóstico , Neoplasias Pulmonares/genética , Proteínas Nucleares/genética , Transdução de Sinais , Estudos Prospectivos , Eletroforese/métodos , Imuno-Histoquímica , Adenocarcinoma/patologia , Transporte Biológico
10.
Rev. esp. med. nucl. imagen mol. (Ed. impr.) ; 36(6): 371-376, nov.-dic. 2017. graf, tab
Artigo em Inglês | IBECS | ID: ibc-167311

RESUMO

Objectives. The preparation of 131I-trazodone hydrochloride and its biological evaluation as a promising brain imaging radiopharmaceutical using two routes of administration. Material and methods. Trazodone (TZ) was radiolabelled with 131I using direct electrophilic substitution, and different factors affecting labelling yield were studied. Quality control of 131I-TZ was carried out using ascending paper chromatography, paper electrophoresis, and high pressure liquid chromatography (HPLC). In vivo biodistribution of 131I-TZ was evaluated in Swiss albino mice using 3 methods: intravenous 131I-TZ solution (IVS), intranasal 131I-TZ solution (INS), and intranasal 131I-TZ microemulsion (INME). Results. Optimum labelling yield of 91.23±2.12% was obtained with in vitro stability of 131I-TZ up to 6h at room temperature. The biodistribution results showed a notably higher and sustained brain uptake for INME compared to IVS and INS at all time intervals. In addition, heart and blood uptake levels for INME were lower than those for IV solution which, in turn, could decrease the systemic side effects of trazodone. Also, the 131I-trazodone INME brain uptake of 6.7±0.5%ID/g was higher than that of 99mTc-ECD and 99mTc-HMPAO (radiopharmaceuticals currently used for brain imaging). Conclusion. 131/123I-trazodone formulated as INME could be used as a promising radiopharmaceutical for brain imaging (AU)


Objetivos. Preparación del hidrocloruro de 131I-Trazodona y su evaluación biológica como radiofármaco prometedor de imagen cerebral utilizando dos vías de administración. Material y métodos. El marcaje de la Trazodona (TZ) con 131I se realizó mediante sustitución electrofílica directa y se estudiaron diferentes factores que pueden afectar al rendimiento del marcaje. El control de calidad de 131I-TZ se realizó usando cromatografía en papel ascendente, electroforesis en papel y HPLC. La biodistribución in vivo de 131I-TZ se evaluó en ratones albinos suizos de tres maneras: solución intravenosa de 131I-TZ (IV), solución intranasal de 131I-TZ (IN) y microemulsión intranasal 131I-TZ (MEIN). Resultados. Se obtuvo un rendimiento de marcaje óptimo de 91,23±2,12% con una estabilidad in vitro de 131I-TZ hasta 6 horas a temperatura ambiente. Los resultados de biodistribución mostraron una captación cerebral elevada y mayor para la MEIN en comparación con la solución IV e IN en todos los intervalos de tiempo. Además, la captación vascular y cardíaca para la MEIN fueron más bajos que los de la solución IV, lo que disminuiría los efectos secundarios sistémicos de la TZ. La captación cerebral de 131I-TZ MEIN fue de 6,7±0,5% ID/g, siendo mayor que la de 99mTc-ECD y 99mTc-HMPAO (radiofármacos utilizados actualmente para la imagen cerebral). Conclusión. El 131/123I-TZ formulado como MEIN podría ser utilizado como un radiofármaco prometedor para la imagen cerebral (AU)


Assuntos
Animais , Camundongos , Radioisótopos do Iodo/administração & dosagem , Compostos Radiofarmacêuticos/administração & dosagem , Técnicas In Vitro/métodos , Trazodona/administração & dosagem , Administração Intranasal , Eletroforese , Cromatografia , Cérebro/efeitos da radiação
11.
Rev. lab. clín ; 10(3): 129-138, jul.-sept. 2017. tab, ilus
Artigo em Espanhol | IBECS | ID: ibc-164880

RESUMO

El tracto genital femenino ejecuta un análisis de semen particular, identificando y seleccionando el mejor espermatozoide para la fecundación. Este análisis in vivo trata de asegurar que la descendencia reciba el material genético de mejor calidad. La selección artificial de espermatozoides implica eliminar las barreras naturales con las que se encuentra el gameto masculino en su largo recorrido. Las técnicas convencionales de selección de espermatozoides que se emplean en reproducción asistida (swim-up y gradientes de densidad específicos), se fundamentan en la recuperación o selección de los espermatozoides con mejor movilidad y morfología, pero no distinguen espermatozoides funcionalmente capaces de fecundar ni genéticamente normales. El objetivo de las técnicas avanzadas es seleccionar el espermatozoide con las mejores características funcionales para ICSI o FIV y conseguir una progenie sana. La introducción de nuevas técnicas en el Laboratorio de Andrología podría suponer un salto cualitativo en los resultados en reproducción (AU)


The female genital tract performs a particular sperm analysis, identifying and selecting the best sperm for fertilization. This in vivo analysis seeks to ensure that the offspring receives the best genetic material. Through artificial sperm selection, natural barriers along the female tract are skipped. Conventional selection techniques used in assisted reproduction are based on sperm motility and morphology, not distinguishing between fecundating ability or genetic normality. The object of advanced sperm selection techniques is to retrieve the spermatozoa with better functional characteristics for intracytoplasmic sperm injection or in vitro fertilization. The introduction of these techniques in the andrology laboratory might represent a qualitative leap in assisted reproduction results (AU)


Assuntos
Humanos , Masculino , Espermatozoides/fisiologia , Contagem de Espermatozoides/tendências , Capacitação Espermática/fisiologia , Motilidade Espermática/fisiologia , Técnicas de Reprodução Assistida/instrumentação , Injeções de Esperma Intracitoplásmicas/instrumentação , Birrefringência , Motilidade Espermática , Injeções de Esperma Intracitoplásmicas , Injeções de Esperma Intracitoplásmicas/métodos , Infertilidade Masculina/diagnóstico , Infertilidade Masculina/patologia , Fertilização , Análise Espectral Raman/instrumentação , Eletroforese/instrumentação , Citometria de Fluxo/instrumentação
12.
Rev. iberoam. micol ; 34(3): 165-170, jul.-sept. 2017. ilus, tab
Artigo em Espanhol | IBECS | ID: ibc-165195

RESUMO

Antecedentes. Candida parapsilosis constituye un complejo de especies formado por Candida parapsilosis sensu stricto, Candida orthopsilosis y Candida metapsilosis. Estudios a nivel mundial han descrito su epidemiología y sensibilidad a los antifúngicos. Objetivos. Los objetivos de este trabajo fueron identificar molecularmente aislamientos de sangre del complejo Candida parapsilosis y determinar su sensibilidad in vitro a los antifúngicos de uso sistémico. Métodos. Se estudiaron 86 aislamientos del complejo C. parapsilosis, provenientes de la Red de Vigilancia de Candidemia del Departamento de Micología del Instituto Nacional de Higiene Rafael Rangel, recogidos en el periodo 2008-2011. Se realizó la amplificación del gen de la deshidrogenasa alcohólica secundaria por reacción en cadena de la polimerasa y los productos fueron analizados mediante los polimorfismos de la longitud de sus fragmentos de restricción, mediante el uso de la enzima BanI. Las pruebas de sensibilidad se realizaron mediante Etest® según las instrucciones del fabricante, con modificaciones. Resultados. De los 86 aislamientos estudiados, 81 (94,2%) fueron C. parapsilosis sensu stricto, 4 (4,6%) C. orthopsilosis y uno (1,2%) C. metapsilosis. C. parapsilosis fue sensible a la anfotericina B y la caspofungina, y su porcentaje de resistencia al fluconazol y el voriconazol fue bajo. C. orthopsilosis y C. metapsilosis fueron sensibles a todos los antifúngicos probados. Conclusiones. Los resultados aportaron por primera vez en Venezuela información importante sobre la distribución del complejo C. parapsilosis en casos de candidemia, y apoyan la necesidad de continuar con los programas de vigilancia, incluyendo la discriminación molecular de las especies y las pruebas de sensibilidad a los antifúngicos, que permitirán orientar la terapia específica (AU)


Background. Candida parapsilosis is a species complex consisting of Candida parapsilosis sensu stricto, Candida orthopsilosis, and Candida metapsilosis. Studies worldwide have described its epidemiology and susceptibility to antifungal agents. Aims. The aims of this study were to carry out the molecular identification of blood isolates belonging to the Candida parapsilosis species complex, and to determine their in vitro susceptibility to antifungals of systemic use. Methods. A study of 86 strains of C. parapsilosis species complex collected in 2008-2011 and obtained from the Candidaemia Surveillance Network of Mycology Department of the Rafael Rangel National Institute of Hygiene, was made. Secondary alcohol-dehydrogenase gene amplification was performed using polymerase chain reaction, and the products were analysed by restriction fragments length polymorphisms using the enzyme BanI. Susceptibility tests were performed using Etest®, following the manufacturer's instructions with modifications. Results. Of the 86 isolates studied, 81 (94.2%) were C. parapsilosis sensu stricto, 4 (4.6%) C. orthopsilosis, and one (1.2%) C. metapsilosis. C. parapsilosis isolates were susceptible to amphotericin B and caspofungin, showing low rates of resistance to fluconazole and voriconazole. C. orthopsilosis and C. metapsilosis were susceptible to all the antifungals tested. Conclusions. The results obtained in Venezuela provide for the first time important information about the distribution of C. parapsilosis species complex in cases of candidaemia, and support the need for continuing surveillance programs, including molecular discrimination of species and antifungal susceptibility tests, which may guide specific therapy (AU)


Assuntos
Humanos , Candida/isolamento & purificação , Antifúngicos/administração & dosagem , Antifúngicos/isolamento & purificação , Antifúngicos/farmacologia , Fluconazol/análise , Voriconazol/análise , Testes de Sensibilidade Microbiana/instrumentação , Sensibilidade e Especificidade , Técnicas In Vitro/métodos , Candidemia/diagnóstico , Reação em Cadeia da Polimerase/métodos , Eletroforese
13.
Rev. lab. clín ; 10(2): 100-104, abr.-jun. 2017. tab, ilus, graf
Artigo em Espanhol | IBECS | ID: ibc-163002

RESUMO

El Mieloma Múltiple (MM) IgD es una entidad compleja que representa menos del 2% de los casos de pacientes con MM. En este estudio describimos las características clínicas y analíticas de una serie de cinco pacientes diagnosticados con MM IgD en nuestro área geográfica destacando la importancia de los hallazgos del laboratorio clínico en el estudio de estos pacientes. La serie de pacientes se caracterizó por una prevalencia del género masculino, con una edad comprendida entre 50 y 83 años y un predominio de la cadena ligera monoclonal lambda. Al diagnóstico, todos los pacientes presentaron fallo renal agudo y lesiones óseas características de la enfermedad (AU)


IgD Multiple Myeloma (MM) is a rare entity that affects less than 2% of patients with MM. The aim of this study is to describe the clinical and analytical findings in five patients diagnosed with IgD MM in our geographic area. Furthermore, the relevance of clinical laboratory findings in the diagnostic protocol for these patients is demonstrated. The majority of patients studied were males, with ages ranging from 50 to 83 years, and a predominance of lambda light chain. At diagnosis, all the patients were shown to have impaired renal function and bone lesions characteristic of the disease (AU)


Assuntos
Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Imunoglobulina D/administração & dosagem , Imunoglobulina D/análise , Mieloma Múltiplo/diagnóstico , Amiloidose/diagnóstico , Paraproteinemias/sangue , Paraproteinemias/diagnóstico , Sobrevivência , Protocolos/métodos , Técnicas de Laboratório Clínico , Cadeias lambda de Imunoglobulina/análise , Estudos Retrospectivos , Eletroforese
14.
Rev. iberoam. micol ; 34(1): 43-45, ene.-mar. 2017. ilus
Artigo em Inglês | IBECS | ID: ibc-160734

RESUMO

Background. No phenotypic methods are available to unequivocally differentiate species within the Candida glabrata complex. Aims. To develop a new multiplex PCR method to differentiate between the three species of the C. glabrata species complex, as well as using it to study a C. glabrata collection to discover strains of the newly described species. Methods. The method was developed based on the Internal Transcribed Spacer (ITS) sequence differences between the species. It was validated by using a blinded collection of strains and, finally, the new molecular method was used to study a collection of 192 C. glabrata species complex strains. The obtained results were compared with ITS sequencing. Results. The proposed method showed 100% concordance with ITS sequencing and proved to be effective for clinical and epidemiological applications. Two Candida bracarensis and three Candida nivariensis were found out of the 192 studied strains (0.93% and 1.40% prevalence, respectively). Conclusions. A fast, inexpensive, robust and highly reproducible multiplex PCR method is presented. Its usefulness is demonstrated by studying a large collection of C. glabrata sensu lato strains (AU)


Antecedentes. No hay métodos fenotípicos disponibles para diferenciar las especies del complejo Candida glabrata. Objetivos. Diseñar un método de PCR multiplex para diferenciar las tres especies del complejo C. glabrata y usarlo para estudiar una colección de cepas identificadas anteriormente como C. glabrata. Métodos. El método fue desarrollado con base en las diferencias de la secuencia internal transcribed spacer (ITS) entre las especies. El método se validó mediante el uso de una colección de cepas incógnitas y se utilizó posteriormente para estudiar una colección de 192 cepas. Los resultados se compararon con las secuencias ITS. Resultados. El método propuesto mostró 100% de concordancia con la secuenciación de las regiones ITS y demostró ser eficaz clínica y epidemiológicamente. Se identificaron dos aislamientos de Candida bracarensis y tres de Candida nivariensis dentro de las 192 cepas identificadas fenotípicamente como C. glabrata (prevalencia de 0,93% y 1,40%, respectivamente). Conclusiones. Presentamos un método de PCR múltiplex rápido, económico y fiable. La utilidad de la metodología queda demostrada con el estudio de una gran colección de cepas de C. glabrata sensu lato (AU)


Assuntos
Reação em Cadeia da Polimerase/instrumentação , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/tendências , Candida glabrata , Candida glabrata/isolamento & purificação , Candida glabrata/efeitos da radiação , Biologia Molecular/métodos , Candida/isolamento & purificação , Candida/efeitos da radiação , Eletroforese/classificação , Eletroforese/métodos , Eletroforese/tendências
16.
Rev. lab. clín ; 10(1): 14-20, ene.-mar. 2017. tab, ilus
Artigo em Espanhol | IBECS | ID: ibc-160832

RESUMO

Introducción. La terapia con anticuerpos monoclonales desarrollada en los últimos años se utiliza actualmente en numerosas patologías. Al ser similares estructuralmente a las inmunoglobulinas endógenas podrían detectarse como bandas monoclonales mediante técnicas de electroforesis e inmunofijación en el análisis de proteínas séricas resultando un falso positivo e informarse erróneamente como componente monoclonal. Objetivo. Reproducir in vitro la concentración sérica de rituximab, infliximab y tocilizumab y evaluar la interferencia producida por estos fármacos en la electroforesis e inmunofijación de proteínas en gel de agarosa. Material y métodos. Se parte de un pool de sueros de pacientes sanos a los que se añade la cantidad de fármaco necesaria para obtener concentraciones decrecientes de anticuerpos monoclonales. Se realiza la electroforesis y la inmunofijación a estas muestras con diluciones de fármacos en el equipo IINTERLAB G26 de Biometa. Resultados y conclusiones. En los tres fármacos el proteinograma resulta positivo para las concentraciones de fármaco más elevadas y la señal va disminuyendo hasta desaparecer según disminuye la concentración de fármaco. Así mismo la inmunofijación también resulta positiva observando una banda que corresponde a inmunoglobulina G kappa con la diferencia que rituximab y tocilizumab migran en una zona más próxima al cátodo que infliximab. Rituximab, infliximab y tocilizumab se detectan como bandas monoclonales en la electroforesis de proteínas e inmunofijaciones correspondientes siendo falsos positivos en la detección de componentes monoclonales. Es necesario que se conozcan los patrones del proteinograma y de la inmunofijación de los anticuerpos monoclonales utilizados en cada hospital con la finalidad de evitar informar falsos positivos (AU)


Introduction. Monoclonal antibody therapy is being used nowadays to treat many diseases. Due to the structural similarity to endogenous immunoglobulins, they might be detected while testing for serum proteins using immunifixation and electrophoresis techniques, giving as a result false positives and being reported as monoclonal components mistakenly. Objectives. To reproduce rituximab, infliximab and tocilizumab serum concentrations in vitro and to evaluate the interference of these drugs when performing protein electrophoresis and immunofixation with agarose gel electrophoresis. Methods and materials. The amounts of drug needed in order to achieve decreasing concentrations of monoclonal antibody were added to sera samples taken from healthy individuals. Both serum protein electrophoresis and inmunofixation were performed into those drug dilutions using IINTERLAB G26 as equipment. Results and conclusions. The electrophoresis was positive for high drug concentrations in the three monoclonal antibody groups, being the signals weaker with decreasing concentrations. Same result was obtained with immunofixation, noting a band corresponding to a kappa G immunoglobulin which migrates closer to the cathode for the cases of rituximab and tocilizumab than for infliximab. It has been proved that rituximab, infliximab and tocilizumab are detected by agarose gel electrophoresis and their corresponding immunofixations and may cause false positives while detecting monoclonal components. Professionals should know the electrophoresis and immunofixation patterns of the monoclonal antibody drug used in their workplaces in order to avoid reporting false positives (AU)


Assuntos
Anticorpos Monoclonais/análise , Anticorpos Monoclonais/isolamento & purificação , Eletroforese/instrumentação , Eletroforese/métodos , Proteínas Sanguíneas/análise , Proteínas Sanguíneas/imunologia , Proteínas Sanguíneas/isolamento & purificação , Técnicas In Vitro/métodos , Técnicas In Vitro , Reações Falso-Positivas , Eletroforese em Gel de Ágar/métodos , Eletroforese em Gel de Ágar , Proteína C-Reativa/análise , Rituximab/imunologia , Rituximab/farmacologia , Infliximab/análise
17.
Rev. lab. clín ; 10(1): 49-54, ene.-mar. 2017. tab, ilus, graf
Artigo em Espanhol | IBECS | ID: ibc-160836

RESUMO

La macroglobulinemia de Waldenström es un linfoma linfoplasmocítico caracterizado por una proliferación monoclonal de linfocitos B productores de inmunoglobulina M que infiltran la médula ósea. Las manifestaciones neurológicas asociadas a la macroglobulinemia de Waldenström suelen ser debidas al fenómeno de hiperviscosidad o a neuropatías desmielinizantes mediadas por inmunoglobulina M. Cuando la afectación neurológica es debida a la infiltración del sistema nervioso central por las células linfoplasmocitoides, se produce un síndrome denominado de Bing-Neel, con baja prevalencia y variedad de manifestaciones clínicas. Se presenta el caso clínico de una mujer de 76 años con antecedentes de macroglobulinemia de Waldenström, con un cuadro neurológico repentino de alteración del lenguaje y torpeza en la mano derecha. Cabe destacar la relevancia del laboratorio clínico en el diagnóstico del síndrome de Bing-Neel y en el seguimiento del tratamiento (AU)


Waldenström macroglobulinemia is a lymphoplasmacytic lymphoma defined by a monoclonal proliferation of bone marrow infiltrating immunoglobulin M producing B lymphocytes. Neurological simptoms of Waldenström macroglobulinemia are mainly dominated by signs of hyperviscosity and autoimmune neuropathies mediated by immunoglobulin M. Neurological involvement secondary to the infiltration of IgM producing B lymphocytes, is defined as a Bing-Neel syndrome. This syndrome has a low prevalence and the clinical manifestations are variable. The case described is about a 76 year-old female with a history of Waldenström macroglobulinemia, who presents sudden neurological signs such as alteration of spoken language and clumsiness of the right hand. The clinical laboratory has a primary role in the diagnosis of Bing-Neel syndrome and monitoring of the treatment (AU)


Assuntos
Humanos , Feminino , Idoso , Transtornos Linfoproliferativos/complicações , Transtornos Linfoproliferativos/diagnóstico , Transtornos Linfoproliferativos/patologia , Imunoglobulina M/análise , Técnicas de Diagnóstico Neurológico/instrumentação , Imagem por Ressonância Magnética/métodos , Espectroscopia de Ressonância Magnética/métodos , Eletroforese/métodos , Serviços de Laboratório Clínico/normas , Serviços de Laboratório Clínico , Eletroforese das Proteínas Sanguíneas/tendências , Carcinomatose Meníngea/diagnóstico , Carcinomatose Meníngea/patologia
19.
Clin. transl. oncol. (Print) ; 18(8): 749-759, ago. 2016. tab, ilus
Artigo em Inglês | IBECS | ID: ibc-154049

RESUMO

The primary cause of tumor-related death in breast cancer is still represented by distant metastasization. The dissemination of tumor cells from the primary tumor to distant sites through bloodstream cannot be early detected by standard imaging methods. Circulating tumor cells (CTCs) play a major role in the metastatic spread of breast cancer. Different analytical systems for CTCs isolation and detection have been developed and novel areas of research are directed towards developing assays for CTCs molecular characterization. This review describes the current state of art on CTCs detection techniques and the present and future clinical implications of CTCs enumeration and characterization (AU)


No disponible


Assuntos
Humanos , Feminino , Neoplasias da Mama/diagnóstico , Células Neoplásicas Circulantes , Células Neoplásicas Circulantes/patologia , Biomarcadores Tumorais/análise , Micropeneiramento/métodos , Eletroforese/instrumentação , Eletroforese , Imuno-Histoquímica/instrumentação , Imuno-Histoquímica/métodos , Imuno-Histoquímica , Citometria de Fluxo/métodos , Técnicas de Amplificação de Ácido Nucleico
20.
Clin. transl. oncol. (Print) ; 18(8): 782-791, ago. 2016.
Artigo em Inglês | IBECS | ID: ibc-154053

RESUMO

Purpose: ZFP36 ring finger protein (ZFP36) and the suppressor of cytokine signaling 3 (SOCS3) have been reported to, respectively, regulate NF-jB and STAT3 signaling pathways. To better understand the correlation of NF-jB and STAT3 negative regulates pathway, we have investigated the involvement of ZFP36 and SOCS3 expressions in human prostate cancer (PCa). Methods: In the present study, paired patient tissue microarrays were analyzed by immunohistochemistry, and the ZFP36 protein expression was quantitated as immunoreactive scores in patients with PCa. Associations between ZFP36/SOCS3 expression and various clinicopathological features and prognosis of PCa patients were statistically analyzed based on the Taylor database. Then, the functions of ZFP36 and SOCS3 in cancerous inflammation were determined using qPCR and immunohistochemistry in vitro and in vivo. Results: ZFP36 protein expression in PCa tissues was significantly lower than those in non-cancerous prostate tissues (P < 0.05). In mRNA level, ZFP36 and SOCS3 had a close correlation with each other (P < 0.01, Pearson r = 0.848), and its upregulation was both significantly associated with low Gleason score (P < 0.001 and P < 0.001, respectively), negative metastasis (P < 0.001 and P < 0.001, respectively), favorable overall survival (P < 0.001 and P < 0.05, respectively), and negative biochemical recurrence (P < 0.001 and P < 0.001, respectively). Functionally, LPS treatment could lead to the overexpression of ZFP36 and SOCS3 in vitro and vivo. Conclusions: Our data offer the convincing evidence for the first time that the aberrant expressions of ZFP36 and SOCS3 may be involved into the progression and patients’ prognosis of PCa, implying their potentials as candidate markers of this cancer


No disponible


Assuntos
Humanos , Masculino , Proteínas Nucleares/análise , Neoplasias da Próstata/diagnóstico , Tristetraprolina/análise , Proteínas Supressoras da Sinalização de Citocina/análise , Proteínas Supressoras da Sinalização de Citocina/isolamento & purificação , Prognóstico , Proteínas Nucleares/genética , Imuno-Histoquímica/instrumentação , Imuno-Histoquímica , Inflamação/complicações , Inflamação/diagnóstico , RNA/análise , Estimativa de Kaplan-Meier , Análise Multivariada , Eletroforese/métodos
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