RESUMO
PURPOSE: This study aimed to investigate the correlation between calculated globulin (CG, total protein level minus albumin level) and the gamma globulin fraction (Gamma), obtained from serum protein electrophoresis with serum IgG levels in adults (≥ 18 years). METHODS: Using linear regression models, analyses of CG and Gamma levels correlation with IgG levels in adults were performed. Receiver-operator curves were created to determine cutoff values and the respective sensitivity and specificity measures. RESULTS: A total of 886 samples were analyzed. CG and Gamma were positively and statistically correlated with IgG levels (r2 = 0.4628 for CG, and = 0.7941 for Gamma, p < 0.0001 for both analyses). For the detection of hypogammaglobulinemia, i.e., IgG level below the reference value (6 g/L), a CG cutoff value of 24 g/L showed a sensitivity of 86.2% (95% CI 69.4-94.5) and a specificity of 92% (90.0-93.6). A Gamma cutoff value of 7.15 g/L yielded a sensitivity of 100% (88.3-100) and a specificity of 96.8 (95.3-97.8). CONCLUSION: Both CG and Gamma levels determined by protein electrophoresis analysis may be used to screen for antibody deficiencies in adults, enabling earlier diagnosis of antibody deficiencies in a routine clinical setting.
Assuntos
Agamaglobulinemia , Doenças da Imunodeficiência Primária , Humanos , Adulto , Eletroforese , Globinas , Imunoglobulina GRESUMO
BACKGROUND: Common variable immunodeficiency (CVID) is a rare disease that affects children and adults and is often difficult to diagnose. Despite being one of the most frequent causes of immunodeficiency, involving gastrointestinal (GI), respiratory, and hematological systems, the disease onset can have heterogeneous and intermittent symptoms, frequently leading to diagnostic delay. GI symptoms are common and can include diarrhea, but the asymptomatic periods lead to overlooking the recurrent pattern. The same can occur with respiratory infections, thus delaying CVID suspicion. The starting point for CVID diagnosis is the decreased gamma globulin levels in serum protein electrophoresis (SPE), also observed through direct immunoglobulin's dosage. CASE PRESENTATION: The patient is a 38 years-old man who had intermittent diarrhea and recurrent airway infections for 19 years, but the CVID diagnosis was achieved only after SPE was carried out. At that time, he was already malnourished, and developed other complications related to CVID in a short period. CONCLUSIONS: SPE is readily available and inexpensive, but is not part of the laboratory approach in diarrhea. According to the case presented herein, it can be useful for patients with recurrent infections or other clues of the disease.
Assuntos
Imunodeficiência de Variável Comum , Diagnóstico Tardio , Adulto , Proteínas Sanguíneas , Imunodeficiência de Variável Comum/complicações , Imunodeficiência de Variável Comum/diagnóstico , Diagnóstico Tardio/efeitos adversos , Diarreia/etiologia , Eletroforese , Humanos , MasculinoRESUMO
RESUMEN Fundamento: La electroforesis de proteínas y las cadenas ligeras libres en suero son técnicas utilizadas en el diagnóstico del mieloma múltiple. Sin embargo, la utilidad diagnóstica de ambas pruebas puede variar según el método empleado y condiciones reales del medio donde se realicen. Objetivo: Determinar el valor diagnóstico de la electroforesis de proteínas y de las cadenas ligeras libres en suero en el mieloma múltiple. Metodología: Se realizó un estudio retrospectivo de los parámetros electroforesis de proteínas en suero y cadenas ligeras libres en suero a 43 pacientes con diagnóstico de mieloma múltiple por evaluación de la médula ósea. La electroforesis de proteínas se realizó por el método convencional de separación de proteínas sobre papel de acetato de celulosa y para las cadenas ligeras libres se aplicó un ensayo inmunoturbidimétrico en el que se usó un analizador químico (Cobas 311). Se calcularon 7 parámetros que evaluaron la exactitud diagnóstica. Resultados: Todos los parámetros que evaluaron la exactitud diagnóstica estuvieron dentro de los intervalos de confianza en ambas pruebas. Conclusiones: La electroforesis de proteínas y las cadenas ligeras libres en suero son ensayos de gran utilidad en el diagnóstico del mieloma múltiple y se deben utilizar en conjunto para la mayor captación posible de casos.
ABSTRACT Background: Protein electrophoresis and serum free light chains are techniques used in the diagnosis of multiple myeloma. However, the diagnostic utility of both tests may vary according to the method used and the actual conditions of the environment where they are performed. Objective: To determine the diagnostic value of protein electrophoresis and serum free light chains in multiple myeloma. Methodology: A retrospective study of serum protein electrophoresis parameters and serum free light chains was conducted in 43 patients diagnosed with multiple myeloma by bone marrow evaluation. Protein electrophoresis was completed by the conventional method of protein separation on cellulose acetate paper and for free light chains an immunoturbidimetric assay was applied in which a chemical analyzer (Cobas 311) was used. Seven parameters were calculated to evaluate diagnostic accuracy. Results: All parameters assessing diagnostic accuracy were within confidence intervals in both tests. Conclusions: Protein electrophoresis and serum free light chains are very useful assays in the diagnosis of multiple myeloma and should be used in conjunction for the highest possible approval of cases.
Assuntos
Eletroforese das Proteínas Sanguíneas , Cadeias kappa de Imunoglobulina , Eletroforese em Acetato de Celulose , Confiabilidade dos Dados , Mieloma Múltiplo/diagnósticoRESUMO
RESUMEN Introducción: lesión típica ocasionada por el Staphylococcus aureus es el furúnculo o cualquier otro absceso localizado. Objetivo: describir el comportamiento del proteinograma y los niveles de inmunoglobulinas séricas. Métodos: se realizó un estudio longitudinal prospectivo con 70 pacientes portadores de forúnculos infectados por Staphylococcus aureus, que acudieron a las consultas de Inmunología en varias localidades de Villa Clara. Resultados: se determinaron las fracciones proteicas por método de elusión; se cuantificaron las inmunoglobulinas séricas por inmunodifusión radial simple, según edad, sexo y color de la piel de los pacientes. Se contrastaron las variables bajo la prueba de Ji cuadrado, con una significación de confianza del 95 %. Predominaron los pacientes con el color de la piel blanca sobre los no blancos. En la electroforesis de proteínas se obtuvieron resultados normales para las proteínas totales y la fracción gamma. Para la albúmina, fracción alfa 1, alfa 2 y beta globulina se obtuvieron valores bajos, por encima del 95 % válido. Conclusiones: todas las inmunoglobulinas resultaron normales o altas, según los intervalos de referencia para cada grupo de edad. Al correlacionar los valores de las inmunoglobulinas con las fracciones de la electroforesis, fue llamativo el resultado obtenido en la correlación entre la IgA y la fracción beta. De manera general, los anticuerpos no mostraron variaciones significativas en sus correlaciones, lo cual evidenció un pobre papel en la infección. Se concluyó que los valores de alfa 1, alfa 2 y beta globulina pueden tener importancia en la enfermedad.
ABSTRACT Introduction: the typical lesion caused by Staphylococcus aureus is the furuncle or any other localized abscess. Objective: to describe the manifestation of the proteinogram and serum immunoglobulin levels. Methods: a prospective longitudinal study was carried out in 70 patients with furuncles infected by Staphylococcus aureus, who came to the Immunology consultations from various locations of Villa Clara. Results: protein fractions were determined by elution method; serum immunoglobulins were quantified according to age, gender and skin color of the patients by simple radial immunodiffusion. Variables were contrasted under the Chi-square test, with 95% confidence significance. White patients predominated over non-white ones. Normal results were obtained for total proteins and gamma fraction in protein electrophoresis. Low values were obtained for albumin, alpha 1, alpha 2 and beta globulin fraction, above 95% valid. Conclusions: all immunoglobulins were normal or high, according to the reference intervals for each age group. The result obtained in the correlation between IgA and the beta fraction was striking when correlating the immunoglobulin values with the electrophoresis fractions. In general, the antibodies did not show significant variations in their correlations, which evidenced a poor role in infection. We concluded that alpha 1, alpha 2 and beta globulin values may be important in the disease.
Assuntos
Furunculose , Staphylococcus aureus , Eletroforese das Proteínas Sanguíneas , Infecções Cutâneas EstafilocócicasRESUMO
Background: Patients with antibody deficiency may experience exceptionally long diagnostic delays, increasing the risk of life-threatening infections, end-organ damage, mortality, and health costs. Objective: This study aimed to analyze serum protein electrophoresis and verify the correlation between calculated globulin (CG, total protein minus albumin levels) or electrophoretically determined serum gamma globulin fraction (Gamma) with IgG levels in children and adolescents under 18 years old (yo). Methods: We analyzed serum protein electrophoresis (GC or Gamma) and IgG levels from 1215 children and adolescents under 18 yo, classified into 5 age groups. We verified the correlation between CG or Gamma with serum IgG levels. Results: Serum IgG levels varied according to age groups (from 4.3 ± 2.3 g/l in children under 6 months old to 11.4 ± 3.2 g/l in adolescents in the 10-<18 yo group). CG sensitivity and specificity to detect IgG below the reference range for all patients were 93.1% and 81.8%, respectively, and varied according to age group. Gamma sensitivity and specificity for all patients were 100% and 87.8%, respectively, and varied according to age group as well. We found serum IgG levels below the age reference level in 29 patients (2.4% of the cases) using CG or Gamma levels. Conclusion: Both CG and Gamma levels may be of utility as a screening tool for earlier diagnosis of antibody deficiency in children and adolescents under 18 yo.
Assuntos
Anticorpos/sangue , Eletroforese das Proteínas Sanguíneas , Disgamaglobulinemia/diagnóstico , Programas de Rastreamento/métodos , Adolescente , Fatores Etários , Área Sob a Curva , Brasil/epidemiologia , Criança , Pré-Escolar , Disgamaglobulinemia/sangue , Disgamaglobulinemia/epidemiologia , Disgamaglobulinemia/imunologia , Feminino , Humanos , Deficiência de IgA/sangue , Deficiência de IgA/diagnóstico , Deficiência de IgG/sangue , Deficiência de IgG/diagnóstico , Imunoglobulina M/sangue , Imunoglobulina M/deficiência , Lactente , Recém-Nascido , Masculino , Curva ROC , Soroglobulinas/análiseRESUMO
Here we described a case of an asymptomatic 73 years-old female patient in geriatric routine consultation, whose laboratory testing showed hyperproteinemia with accompanying hyperglobulinemia. A diagnosis of BGUS was made only after a correlation among SPEP, densitometry tracing and IFE results was established, evidencing a second peak, that was less evident and not reported at first. These biclonal conditions are of very low incidence in the clinical laboratory, requiring the laboratory professional to have particular skills for their identification. As far as is known, clinical findings in BGUS are similar to those found in MGUS. However, they remain not well understood. Therefore, for an accurate diagnosis of BGUS, the clinical laboratory technician must be trained and sensitized to detect a second M - protein as a band or peak; taking in mind the possible different scenarios in heavy and light chain typing.
Se describe el caso de paciente asintomática de 73 años de edad en consulta geriátrica de rutina, cuyos estudios de laboratorios muestran hiperproteinemia acompañada de hiperglobulinemia. Se estableció el diagnóstico de GBSI después de correlacionar entre resultados de electroforesis de proteínas, trazo de densitometría e inmunofijación en suero, los cuales evidenciaron un segundo pico monoclonal menos evidente y no reportado de primera instancia. Este tipo de condiciones biclonales son de muy baja incidencia en laboratorio clínico, lo cual requiere que profesional de laboratorio tenga ciertas habilidades para su identificación. Hasta donde se conoce, los hallazgos clínicos de GBSI son similares a aquellos encontrados en GMSI. Sin embargo, continúan sin ser bien comprendidas. Por tanto, a fin de un diagnóstico más preciso, el técnico de laboratorio debe estar entrenado y sensibilizado para encontrar una segunda proteína M como banda o pico, tomando en cuenta los diferentes posibles escenarios en la tipificación de cadenas pesadas y ligeras.
RESUMO
Hematological neoplasms are tumors of cells in different states of maturation and differentiation. Since monoclonal gammopathies (MG) refer to B mature lymphocyte neoplasms, lymphogenesis should be well known. We must keep in mind that the last stage of maturation of these lymphocytes is the plasma cell. This is how a MG could appear in the context of a plasma cell neoplasm, such as multiple myeloma or amyloidosis, but also in relation to a lymphoma. A monoclonal peak is produced by mature B lymphocytes or plasma cells that secrete a monoclonal protein (Immunoglobulin), and represents a MG. But it must be emphasized that, in the correct clinical context, a hypogammaglobulinemia can represent a MG as well. Another important point is the understanding and interpretation of requested tests, such as protein electrophoresis (PEP), immunofixation (IFx) or serum free light chains (sFLC). The current MG screening panel includes these three studies (PEF, IFx, sFLC), although a simpler panel measuring PEF and sFLC has also been proposed, but not yet formally validated. Therefore, screening done only with PEP is insufficient.
Assuntos
Humanos , Paraproteinemias/sangue , Paraproteínas/análise , Neoplasias de Plasmócitos/sangue , Paraproteinemias/diagnóstico , Eletroforese das Proteínas Sanguíneas/métodos , Linfócitos B/metabolismo , Neoplasias de Plasmócitos/diagnósticoRESUMO
Proteome analysis represents a promising approach for plant tissue culture since it is now possible to identify and quantify proteins on a large scale. Biomarker discovery and the study of the molecular events associated with in vitro plant morphogenesis are considered potential targets for application of proteomics technologies. This chapter describes a protocol for application in in vitro plant material using two proteomics approaches: 2-DE coupled to mass spectrometry and liquid chromatography-linked tandem mass spectrometry.
Assuntos
Desenvolvimento Vegetal/genética , Proteômica/métodos , Biologia Computacional , Eletroforese em Gel Bidimensional , Espectrometria de Massas , Proteínas de Plantas/isolamento & purificaçãoRESUMO
ABSTRACT We report the first case of hemoglobin SD-Punjab disease, a rare form of sickle-cell disease, in the state of Bahia. Detection was possible by a test for the identification of hemoglobin (Hb) variants with the high-resolution liquid chromatography technique. By means of the molecular study of chromosomal polymorphism with beta-globin S gene, the Bantu haplotype was observed. According to studies, there is strong association between the prevalence of Bantu haplotype and reduced levels of fetal Hb and Hb D-Punjab as a stimulating factor for S polymerization, what contributes to the hematological disorders of the disease and organ damage, as gallstones and aseptic necrosis of the femoral head.
RESUMO Reportamos o primeiro caso de hemoglobinopatia SD-Punjab, uma forma rara da doença falciforme, no estado da Bahia. A detecção ocorreu pelo exame para identificação de hemoglobinas (Hb) variantes com técnica de cromatografia líquida de alta resolução. Através do estudo molecular do polimorfismo no cromossomo com gene da betaglobina S, verificou-se a presença do haplótipo Bantu. Segundo estudos, existe forte associação de prevalência do haplótipo Bantu e níveis reduzidos da Hb fetal e da Hb D-Punjab como fator de estímulo à polimerização da S, o que contribui para os distúrbios hematológicos da doença e a lesão de órgãos, como cálculos biliares e necrose asséptica de cabeça de fêmur.
RESUMO
El proteinograma por electroforesis (PxE) sérico es solicitado para detectar modificaciones del perfil proteico. El objetivo del trabajo fue evaluar las alteraciones de la zona gammaglobulina y su correspondencia con distintos estados clínico-patológicos. Se incluyeron 7.259 pacientes (1-89 años) a los que en 2013 se les solicitó PxE. Según el trazado densitométrico, en la zona gammaglobulina se reconocieron diferentes grupos: hipogammaglobulinemia (<0,60 g/dL), hipergammaglobulinemia policlonal (≥1,80 g/dL), banda monoclonal (BM) y bandas oligoclonales. Prevaleció la hipergammaglobulinemia policlonal (4,2%), seguida por BM (1,4%) e hipogammaglobulinemia (0,8%). Hipergammaglobulinemia policlonal (>3 g/dL) se observó en: hepatitis autoinmune, cirrosis, síndrome de Sjögren, enfermedad mixta del tejido conectivo, HIV, hepatitis C y enfermedad de Castleman. El hallazgo de BM correspondió a 47% de pacientes con gammapatía monoclonal de significado incierto y 40% con mieloma múltiple; el 0,5% fueron casos nuevos. Con hipogammaglobulinemias, en adultos prevaleció la inmunosupresión terapéutica (55%), seguida por diabetes/síndrome metabólico/hipotiroidismo (23%); en niños, 22% por inmunosupresión y 78% con hipogammaglobulinemia no clasificada como inmunodeficiencia primaria. Se concluye que en 6,4% de los PxE se observó alteración de la zona gammaglobulina; prevaleció la hipergammaglobulinemia policlonal. En 1 de cada 200 PxE se pesquisó un paciente con BM. El hallazgo de hipergammaglobulinemia policlonal o BM se correspondió con distintos estados clínico-patológicos.
Serum protein electrophoresis (PEP) is requested to screen changes in the protein profile. The aim of this study was to evaluate alterations in the gamma globulin zone and correspondence with various clinical and pathological states. 7259 patients were included (1-89 years of age) who had been requested a PEP in 2013. According to the densitometric tracing, in the gamma globulin zone different groups were recognized: hypogammaglobulinemia (<0.60 g/dL), polyclonal hypergammaglobulinemia (≥1,80 g/dL), monoclonal band (MB) and oligoclonal band. The polyclonal hypergammaglobulinemia prevailed (4.2%), followed by MB (1.4%) and hypogammaglobulinemia (0.8%). Polyclonal hypergammaglobulinemia (>3 g/dL) was observed in autoimmune hepatitis, alcoholic cirrhosis, Sjögren's syndrome, mixed connective tissue disease, HIV, hepatitis C and Castleman's disease. The MB finding corresponded to a 47% of patients with monoclonal gammopathy of undetermined significance and 40% with multiple myeloma; 0.5% were new cases. In adults, hipogammaglobulinemias prevailed in therapeutic immunosuppression cases (55%), followed by patients with diabetes/ metabolic syndrome/ hypothyroidism (23%); in children, 22% with immunosuppression and 78% corresponded to hipogammaglobulinemias not classified as primary immunodeficiency. To conclude, an alteration in the gamma globulin zone was observed in 6.4% of PEP. In 1 out of 200 PEP MB was found. The finding of polyclonal hypergammaglobulinemia or MB corresponded to different clinicopathological states.
O proteinograma por eletroforese (PXE) sérico é solicitado para detectar modificações no perfil proteíco. O objetivo do trabalho foi avaliar as alterações da área gammaglobulina e sua correspondência com diversos estados clínico-patológicos. Incluíram-se 7259 pacientes (1-89 anos) aos quais, em 2013, foi solicitado um PxE. De acordo com o traçado densitométrico, na área gammaglobulina, diferente grupos foram reconhecidos: hipogammaglobulinemia (<0,60 g/dL), hipergammaglobulinemia policlonal (≥1,80 g/dL), banda monoclonal (BM) e bandas oligoclonais. Prevaleceu a hipergammaglobulinemia policlonal (4,2%), seguida por BM (1,4%) e hipogammaglobulinemia (0,8%). Hipergammaglobulinemia policlonal (>3 g/dL) foi observada em: Hepatite autoimune, cirrose, síndrome de Sjögren, doença mista do tecido conjuntivo, HIV, hepatite C e doença de Castleman. O achado de BM correspondeu a 47% de pacientes com gammapatia monoclonal de significado indeterminado e 40% com mieloma múltiplo; 0,5% eram casos novos. Com hipogammaglobulinemias em adultos prevaleceu a imunossupressão terapêutica (55%), seguida por diabete/síndrome metabólica/hipotireoidismo (23%); em crianças, 22% por imunossupressão e 78% com hipogammaglobulinemia não classificados como imunodeficiência primária. Conclui-se que em 6,4% dos PxE foi observada alteração da área gammaglobulina; prevaleceu a hipergammaglobulinemia policlonal. Em 1 de cada 200 PxE foi encontrado um paciente com BM. O achado de hipergammaglobulinemia policlonal ou BM se correspondeu com diferentes estados clínico-patológicos.
Assuntos
Humanos , Lactente , Pré-Escolar , Criança , Adolescente , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , gama-Globulinas/análise , Eletroforese/métodos , gama-Globulinas , Eletroforese em Gel de Ágar , Hipergamaglobulinemia/patologiaRESUMO
This case study presents a patient with multiple myeloma whose serum specimen exhibits 2 distinct bands in serum protein electrophoresis but only one band in immunofixation electrophoresis. This latter, single band corresponds to the M-spike. An investigation is presented to determine the identity of this disappearing or phantom band. Furthermore, this case is used as a teaching point to explain the criteria used for staging multiple myeloma, how a cell can become a myeloma propagating cell, methods that can be used to identify unexpected bands in serum protein electrophoresis, possible explanations for bands in the beta region, the usual treatment regimens in multiple myeloma and finally specimen collecting and handling procedures for serum protein electrophoresis.
Assuntos
Proteínas Sanguíneas/análise , Mieloma Múltiplo/sangue , Mieloma Múltiplo/diagnóstico , Densitometria , Eletroforese , Citometria de Fluxo , Humanos , Imunoeletroforese , Masculino , Pessoa de Meia-IdadeRESUMO
ABSTRACT Introduction: Radiation induces acute and late alterations in blood proteins. Objective: The present study aims at analyzing in time kinetics the electrophoretic profiles of expression bands of blood protein, in the molecular weight range greater than and equal to that of albumin, modulated by ionizing radiation in the cardiac territory, in animal model. Material and methods: Animals were exposed to a whole-body dose of 5 Gy ionizing radiation (Co-60). Serum samples were collected from isogenic Wistar rats (control and irradiated groups). At a time kinetics of 12, 24, 48, 72, 96 hours and 35 days post-irradiation, thoracolaparotomies were performed with anesthesia, and 0.3 ml of blood was collected between the left ventricle and the pulmonary artery. The samples were held in heparin and their components were separated by centrifugation. Protein bands with similar molecular weight were identified by vertical 10% electrophoresis, silver staining. Results: The findings indicate a systemic acute altered expression of proteins with molecular weight greater than or equal to that of albumin in acrylamide gel, presenting suppression and increased expression due to modulation of preexisting bands, identified in time kinetics. Conclusion: These findings point out to acute alterations of protein expression modulated in time, but also to a late modulation of gene expression.
RESUMO Introdução: A radiação de corpo inteiro induz alterações agudas e tardias no perfil proteico sanguíneo. Objetivo: O presente estudo tem como escopo analisar em cinética de tempo o perfil eletroforético de bandas de expressão das proteínas solúveis do sangue, na faixa de peso molecular superior e igual à albumina, moduladas por radiação ionizante no território cardíaco em modelo animal. Materiais e métodos: Animais foram expostos à radiação de Co-60, em dose de 5 Gy corpo inteiro. Foram coletadas amostras de soro sanguíneo em ratos Wistar isogênicos (no grupo-controle e no irradiado). Em uma cinética de tempo de 12, 24, 48, 72 e 96 horas e 35 dias pós-exposição, foram realizadas toracolaparotomias com anestesia profunda e, posteriormente, foi coletado 0,3 ml de sangue entre ventrículo esquerdo e artéria pulmonar. As amostras foram heparinizadas, sendo, em seguida, separados seus componentes por centrifugação. As bandas proteicas de peso molecular similar foram identificadas por eletroforese vertical a 10%, coradas com prata. Resultados: Os achados indicam alteração aguda sistêmica da expressão das proteínas de peso molecular superior ou igual à albumina em gel de acrilamida, representando supressão e aumento de expressões por meio da modulação de bandas preexistentes identificadas em cinética de tempo. Conclusão: Esses achados apontam tanto para uma alteração aguda modulada no tempo da expressão proteica quanto para uma alteração moderada tardia da expressão gênica.
RESUMO
A 37-year-old, female Chilean flamingo (Phoenicopterus chilensis) presented with severe facial angioedema, bilateral corneal and palpebral edema, nictitating membrane paralysis, bradycardia, bradypnea, hypothermia, and numerous stingers and remnants of eastern yellow jacket wasps (Vespula maculifrons) attached to the feathers of the head, palpebrae, and conjunctiva. Evaluation of 2 complete blood cell counts and results of plasma chemical analysis and serum protein electrophoresis revealed severe increases in creatinine phosphokinase and aspartate aminotransferase activity, electrolyte disturbances, and moderate increases in levels of α1, α2, ß1, and γ immunoglobulins when compared with reference interval values and conspecifics. Despite intensive treatment, the bird died 19 hours after presentation. Results of histologic evaluation of tissues were compatible with envenomation. Response to envenomation in avian species is not documented but should be considered in birds presenting with angioedema.
RESUMO
La destruccion del músculo esquelético en la condición patológica conocida como rabdomiolisis resulta en la liberación al torrente sanguíneo de elevadas concentraciones de la proteína mioglobina de 17 kDa, la cual filtra libremente a través del glomérulo sobrepasando frecuentemente la capacidad de reabsorción del túbulo proximal. Por lo tanto, la identificación de mioglobina en orina es una herramienta esencial que complementa otros parámetros bioquímicos en el diagnóstico de la enfermedad. En el presente trabajo, mediante la combinación de electroforesis en geles de agarosa e inmunofijación empleando anticuerpos específicos, se provee evidencia directa de la presencia de mioglobina intacta en la orina de un paciente con insuficiencia renal aguda asociada a rabdomiolisis desencadenada por efecto secundario de una terapia reductora de lípidos. Los datos electroforéticos e inmunoquímicos fueron corroborados mediante secuencia N-terminal de aminoácidos, immunoblot y espectrometría de masa. La simple combinación de electroforesis e inmunofijación provee una estrategia flexible que puede extenderse a la identificación de diversas proteínas involucradas en proteinurias de sobrecarga.(AU)
The destruction of skeletal muscle in a condition known as rhabdomyolysis results in the release into the blood stream of large quantities of the 17 kDa myoglobin that freely filtrates through the glomeruli, often overwhelming the capacity of the proximal tubule for reabsorption. As a result, identification of myoglobin in the urine becomes an essential tool to fully complement other biochemical parameters in the diagnosis of the disease. Using a combination of protein electrophoresis in agarose gels and immunofixation with specific antibodies, direct evidence is provided for the presence of intact myoglobin in the urine of a patient with acute kidney injury due to rhabdomyolysis triggered by undesirable side effects of specific medications. The electrophoretic/immunochemical data was further corroborated by amino acid sequence, Western blot, and mass spectrometry analysis. The simple combination of electrophoresis and immunofixation protocols provides a flexible approach that can be extended to the identification of the various proteins known to be involved in overflow proteinuria.(AU)
A destruiþÒo do músculo esquelético na condiþÒo patológica conhecida como rabdomiólise resulta na liberaþÒo ao torrente sanguíneo de elevadas concentraþ§es da proteína mioglobina de 17 kDa a qual filtra livremente através do glomérulo ultrapassando frequentemente a capacidade de reabsorþÒo do túbulo proximal. Portanto, a identificaþÒo de mioglobina em urina é uma ferramenta essencial que complementa outros parÔmetros bioquímicos no diagnóstico da doenþa. No presente trabalho, mediante a combinaþÒo de eletroforese em géis de agarose e imunofixaþÒo empregando anticorpos específicos, é fornecida evidÛncia direta da presenþa de mioglobina intacta na urina de um paciente com insuficiÛncia renal aguda associada a rabdomiólise desencadeada por efeito secundário de uma terapia redutora de lipídeos. Os dados eletroforéticos e imunoquímicos foram confirmados mediante sequÛncia N-terminal de aminoácidos, immunoblot e espectrometria de massa. A simples combinaþÒo de eletroforese e imunofixaþÒo fornece uma estratégia flexível que pode se estender O identificaþÒo de diversas proteínas envolvidas em proteinúrias de sobrecarga.(AU)
RESUMO
The purpose of this work was to evaluate biochemical and serological methods to characterize and identify Candida species from the oral cavity. The strains used were five Candida species previously identified: C. albicans, C. guilliermondii, C. parapsilosis, C. krusei, C. tropicalis, and Kluyveromyces marxianus, as a negative control. The analyses were conducted through the SDS-PAGE associated with statistical analysis using software, chromogenic medium, and CHROMagar Candida (CA), as a differential medium for the isolation and presumptive identification of clinically important yeasts and an enzyme-linked immunoabsorbent assay (ELISA), using antisera produced against antigens from two C. albicans strains. This method enabled the screening of the three Candida species: C. albicans, C. tropicalis, and C. Krusei, with 100% of specificity. The ELISA using purified immunoglobulin G showed a high level of cross-reaction against protein extracts of Candida species. The SDS-PAGE method allowed the clustering of species-specific isolates using the Simple Matching coefficient, S SM = 1.0. The protein profile analysis by SDS-PAGE increases what is known about the taxonomic relationships among oral yeasts. This methodology showed good reproducibility and allows collection of useful information for numerical analysis on information relevant to clinical application, and epidemiological and systematical studies.
Este trabalho teve o propósito de avaliar métodos bioquímicos e sorológicos para serem aplicados na caracterização e identificação de linhagens do gênero Candida isoladas da cavidade bucal. As cepas empregadas representam cinco espécies de Candida previamente identificadas: C. albicans, C. guilliermondii, C. parapsilosis, C. krusei e C. tropicalis, utilizando como controle negativo Kluyveromyces marxianus. Foram empregadas as técnicas de gel de poliacrilamida (SDS-PAGE) associado à análise estatística em software, CHROMagar Candida (CA), meio cromogênico diferencial descrito para o isolamento e identificação presuntiva de leveduras de importância clínica e um ensaio de imunoabsorção ligado a enzima (ELISA), utilizando antissoro produzido contra extratos protéicos de uma linhagem-padrão de Candida e um isolado de cavidade oral de C. albicans. O método mostrou-se adequado para a identificação presuntiva de C. albicans, C. tropicalis e C. krusei, com 100% de sensibilidade e especificidade, com base na coloração e textura das colônias. O método de ELISA utilizando imunoglobulinas G purificadas apresentou alto teor de reação cruzada com as outras espécies de Candida estudadas. A análise do perfil protéico por SDS-PAGE permitiu agrupar os isolados da cavidade oral por intermédio do coeficiente "Simple Matching", S SM = 1,0. Os perfis protéicos analisados por SDS-PAGE ampliam os conhecimentos sobre as relações taxonômicas de leveduras isoladas da cavidade oral. Esta metodologia demonstra boa reprodutibilidade e origina informações úteis para aplicação clínica e estudos que envolvem a sistemática e a epidemiologia.
RESUMO
The tropical eastern Pacific halfbeak previously considered conspecific with the western Atlantic Hyporhamphus unifasciatus (Ranzani 1842) is described as a new species, H. naos. It resembles H. meeki from the Atlantic and Gulf coasts of the United States in number of gill rakers on the first arch (usually 32-36, mean 33.6), more than in H. unifasciatus (usually 29-32, mean 30.6), but fewer than in other sympatric species of eastern Pacific Hyporhamphus. Results of a three-treatment ANCOVA (H. naos, H. meeki, and H. unifasciatus) show significant differences in slopes and means for all 14 morphometric characters examined, 9 of 14 characters comparing H. naos with H. unifasciatus, and 7 of 14 comparing H. naos with H. meeki. Protein electrophoretic patterns clearly distinguish all three species with a number of fixed allelic differences.
Assuntos
Animais , Beloniformes/classificação , Alelos , Análise de Variância , Beloniformes/genética , Eletroforese , Oceano Pacífico , Proteínas/análiseRESUMO
En este estudio se cuantificaron las inmunoglobulinas G, A y M, las proteínas de fase aguda: alfa 1 antitripsina, haptoglobina y ceruloplasmina, así como las zonas de la electroforesis de proteínas, en 153 niños atletas de alto rendimiento y en 140 niños supuestamente sanos con actividad física normal para su edad, en busca de variaciones en estos parámetros con carácter permanente por la práctica sistemática de intensos ejercicios físicos. De las inmunoglobulinas dosificadas sólo estuvo marcadamente elevada la IgM y disminuida la zona alfa 2 en los niños atletas; sin embargo, no se obtuvieron diferencias en las proteínas de fase aguda dosificadas para ambos grupos, así como en las proteínas totales cuantificadas.
In this study, G, A and M immunoglobulins, the acute phase proteins: alpha 1-antitrypsin, haptoglobin and ceruloplasmin, as well as the zones of protein electrophoresis were quantified in 153 sportschildren of high performance and in 140 apparently sound children with normal physical activity according to their age, in order to look for variations in these parameters with permanent character due to the systematic practice of intensive physical exercises. Of the dosified immunoglobulins only IgM was significantly elevated, whereas the alpha 2 zone was diminished among sportschildren. Differences were found neither in the actue phase proteins dosified for both groups nor in the quantified total proteins.