La citometría de flujo aumenta la proporción de muestras valorables en líquido cefalorraquídeo con recuento celular normal en hemopatías malignas / Flow cytometry increases the proportion of valuable samples in cerebrospinal fluid with normal cell count in malignant blood diseases

INTRODUCCIÓN: El compromiso del líquido cefalorraquídeo (LCR) en hemopatías malignas es un marcador de mal pronóstico y es habitualmente estudiado por citometría de flujo o citología. Ocasionalmente, las muestras de LCR oligocelulares (≤ 5 céls/dL) pueden ser consideradas como no aptas para diagnóstico por la baja cantidad de eventos. Objetivo: Evaluar la proporción de muestras reportadas como valorables para diagnóstico obtenidas por citometría y citología en muestras de LCR oligocelular. Material y Métodos: Se seleccionaron 169 muestras de LCR oligocelular correspondientes a 115 pacientes con hemopatías malignas. Las muestras fueron obtenidas mediante punción lumbar en tubos acondicionados con EDTA y preservante celular (Transfix®). El inmunofenotipo se realizó con panel de 8 colores, 55 (32%) de las cuales se hizo con panel para pequeñas muestras (SST). En todos los casos se incluyó CD14 para identificación de monocitos y CD3 para linfocitos T. La adquisición se realizó en citómetro FACSCantoII® y el análisis en software Infinicyt®. Resultados: La proporción de muestras valorables fue mayor en citometría en comparación con la citología (98% vs 61%, p < 0,000). En la mayoría se identificaron linfocitos T (98%) y/o monocitos (90%). En las muestras con SST, la cantidad de eventos obtenida fue menor en muestras con < de 1 mL (140 vs 556, p < 0,001) y se logró identificar una mediana de 3 poblaciones celulares. Conclusión: La citometría proporciona una mayor cantidad de muestras valorables en los LCR paucicelulares en relación con la citología en muestras de LCR enviadas para estudio de compromiso de LCR por hemopatías malignas.

BACKGROUND: The alteration of cerebrospinal fluid (CSF) in hematologic neoplasms is a poor prognostic marker. The characteristics of CSF are usually analyzed by flow cytometry or cytology. However, paucicellular CSF samples (≤5 cells/dL) can sometimes be considered unsuitable for analysis due to the low number of events. Objective: To evaluate the proportion of samples reported as suitable for analysis obtained by cytometry (FCM) and cytology in paucicellular CSF samples. Material and Methods: 169 samples ofpaucicellular CSF corresponding to 115 patients with hematologic neoplasms were selected. The samples were obtained by lumbar puncture in tubes conditioned with EDTA and Transfix®. We characterized the immunophenotype ofCSF samples with an 8-color panel, and 55 samples (32%) were in a small sample tube (SST). In all cases, monocytes were identified by CD14 labeling and T lymphocytes by CD3 labeling. The acquisition was carried out in a FACSCantoII® cytometer, and the analysis was performed using Infinicyt® software. Results: The proportion of samples suitable for analysis was higher in FCM compared to cytology (98% vs 61%, p < 0.000). We identified the presence of T lymphocytes and/or monocytes in most samples (98% and 90%, respectively). In the SST samples, the number of events recorded in low-volume samples (< 1 mL) was lower than in samples with higher volume (140 vs 556, p < 0.001), with a median of identification of 3 cell populations. Conclusion: FCM allows the analysis of a higher proportion ofpaucicellular CSF samples than cytology in hematologic neoplasms study.

Use of cerebrospinal fluid CXCL13 concentration for diagnosis and monitoring of neurosyphilis: a three-case report

Introduction: Previous retrospective studies have demonstrated that the concentration of chemokine ligand CXCL13 in cerebrospinal fluid (CSF-CXCL13) is a promising biomarker in the diagnosis of neurosyphilis and, additionally, in the monitoring of therapeutic efficacy. Objective: To describe three cases of patients with neurosyphilis (NS) treated at Hospital Universitário Gaffrée e Guinle, in Rio de Janeiro, Brazil, with suspected active syphilis with neurological symptoms. Case report: Three patients from Rio de Janeiro, Brazil, were investigated for symptomatic NS. The concentration of CSF-CXCL13 was prospectively performed by enzyme-linked immunosorbent assay (ELISA) in all participants at baseline and in follow-up visits at 3 months after therapy. CSF-CXCL13 concentrations were significantly higher in all three patients with established NS. The CSF-CXCL13 concentrations decreased after 3 months of therapy compared to baseline in all cases reported. The added high concentration of CSF-CXCL13 plus CSF-TPHA reactivity above 1:40 titer agreed with the diagnosis of NS in 100% of the cases. Conclusion: In this case series, we present three cases of NS diagnosed using CXCL13 in CSF as a complementary test. These case series suggest that the clinical use of CSF-CXCL13 is useful as a supplementary biomarker for NS and for monitoring the effectiveness of NS therapy, especially in patients with nonreactive CSF-VDRL, excluding other neurologic diseases

[Flow cytometry increases the proportion of valuable samples in cerebrospinal fluid with normal cell count in malignant blood diseases]. / La citometría de flujo aumenta la proporción de muestras valorables en líquido cefalorraquídeo con recuento celular normal en hemopatías malignas.

BACKGROUND: The alteration of cerebrospinal fluid (CSF) in hematologic neoplasms is a poor prognostic marker. The characteristics of CSF are usually analyzed by flow cytometry or cytology. However, paucicellular CSF samples (≤5 cells/dL) can sometimes be considered unsuitable for analysis due to the low number of events. OBJECTIVE: To evaluate the proportion of samples reported as suitable for analysis obtained by cytometry (FCM) and cytology in paucicellular CSF samples. MATERIAL AND METHODS: 169 samples ofpaucicellular CSF corresponding to 115 patients with hematologic neoplasms were selected. The samples were obtained by lumbar puncture in tubes conditioned with EDTA and Transfix®. We characterized the immunophenotype ofCSF samples with an 8-color panel, and 55 samples (32%) were in a small sample tube (SST). In all cases, monocytes were identified by CD14 labeling and T lymphocytes by CD3 labeling. The acquisition was carried out in a FACSCantoII® cytometer, and the analysis was performed using Infinicyt® software. RESULTS: The proportion of samples suitable for analysis was higher in FCM compared to cytology (98% vs 61%, p < 0.000). We identified the presence of T lymphocytes and/or monocytes in most samples (98% and 90%, respectively). In the SST samples, the number of events recorded in low-volume samples (< 1 mL) was lower than in samples with higher volume (140 vs 556, p < 0.001), with a median of identification of 3 cell populations. CONCLUSION: FCM allows the analysis of a higher proportion ofpaucicellular CSF samples than cytology in hematologic neoplasms study.

[Values of cerebrospinal fluid cytochemistry in pediatric patients with ventriculoperitoneal shunt]. / Valores del análisis citoquímico de líquido cefalorraquídeo en pacientes con derivación ventrículo-peritoneal.

BACKGROUND: The ventriculo-peritoneal shunt (VPS) is the treatment for hydrocephalus, the cerebrospinal fluid (CSF) is evaluated for the management of its complications; however, information on the values of the cytochemistry in this population is insufficient. AIM: To describe the characteristics of the CSF cytochemistry of children in VPS management. METHODS: Descriptive observational study, developed in Bogotá (Colombia), from 2008 to 2016. VPS and related procedures records were reviewed. Patients between 6 months and 18 years were included. RESULTS: A total of 285 records were reviewed, 31 samples were entered. The CSF values were, respectively, for the median and 90% percentile: total leukocytes: 0 and 7 cells/mm3, neutrophils: 0 and 6.8 cells/mm3, lymphocytes: 0 and 2 cells/mm3, proteins: 13.4 and 67.2 mg/dL, glucose: 59 and 27.4 mg/dL. DISCUSSION: Glucose values evinced a normal rank towards the widest inferior limit with protein values exceeding the values expected. Cellularity is the variable with the lowest variation. CONCLUSIONS: The values of the CSF cytochemistry in patients with VPS are not comparable to those of the healthy population and should be interpreted according to the characteristics of this population.

Análise do líquido cérebro-espinhal em cães e gatos com afecções neurológicas / Cerebrospinal fluid analysis in dogs and cats with neurological diseases

Background: Cerebrospinal fluid (CSF) analysis is of great importance for evaluation of patients with central nervoussystem problems, because it helps to confirm or exclude many diseases when evaluated together with other relevant clinical and ancillary laboratory tests. As there are few studies in Brazil describing results of CSF analysis in dogs and cats,the objective of the present study was to evaluate results of CSF analysis in a period of ten years, grouping the changesobserved, mainly total and differential cell count and total protein concentration, according to the following etiologicalcategories of diseases: inflammatory, degenerative, traumatic, neoplastic and vascular disorders.Materials, Methods & Results: Medical records of dogs and cats with neurological diseases that had CSF collected andanalyzed at a Veterinary Teaching Hospital between 2008 and 2017 were retrospectively reviewed. Patient data includedbreed, sex, body weight, age at presentation, findings of clinical and neurological examination, results of CSF analysis,diagnostic imaging and other laboratory tests results including PCR and serologies for infectious diseases. A total of 245CSF exams were studied (240 dogs and five cats). The established normal range for protein concentration was less than25 mg/dL for cisternal collection and less than 45 mg/dL for lumbar puncture. Total nucleated cell count was considerednormal when less than 5 cells/µL, with a predominance of mononuclear cells. Both pleocytosis and increased concentrationof the protein level were classified as discrete, moderate and marked, and the pleocytosis was further classified as mononuclear, neutrophilic and mixed cell pleocytosis. The information obtained was analyzed in relation to the neurologicalsyndromes and etiological categories of neurological...(AU)

Valores del análisis citoquímico de líquido cefalorraquídeo en pacientes con derivación ventrículo-peritoneal / Values of cerebrospinal fluid cytochemistry in pediatric patients with ventriculoperitoneal shunt

Resumen Introducción: La derivación ventrículo-peritoneal (DVP) es el tratamiento para la hidrocefalia. El líquido cefalorraquídeo (LCR) se evalúa para el manejo de sus complicaciones; sin embargo, la información de los valores del citoquímico en esta población es insuficiente. Objetivo: Describir las características del citoquímico del LCR de niños en manejo con DVP. Materiales y Métodos: Estudio de tipo observacional descriptivo, desarrollado en Bogotá (Colombia), entre el año 2008 y 2016. Se revisaron los registros de procedimientos de DVP y relacionados. Se incluyeron pacientes entre 6 meses y 18 años de edad. Resultados: Se revisaron 285 registros e ingresaron 31 muestras. Los valores de LCR fueron, respectivamente, para la mediana y al percentil 90%: leucocitos totales: 0 y 7 céls/mm3, neutrófilos: 0 y 6,8 céls/mm3, linfocitos: 0 y 2 céls/mm3, proteínas: 13,4 y 67,2 mg/dL, glucosa: 59 y 27,4 mg/dL. Discusión: Los valores de glucosa presentan un rango normal hacia el extremo inferior más amplio, con valores de proteínas mayores a los valores esperados. El rango de celularidad es la variable que presenta menor variación. Conclusiones: Los valores del citoquímico de LCR en paciente con DVP no son equiparables a los de la población sana y deben interpretarse según las características propias de esta población.

Background: The ventriculo-peritoneal shunt (VPS) is the treatment for hydrocephalus, the cerebrospinal fluid (CSF) is evaluated for the management of its complications; however, information on the values of the cytochemistry in this population is insufficient. Aim: To describe the characteristics of the CSF cytochemistry of children in VPS management. Methods: Descriptive observational study, developed in Bogotá (Colombia), from 2008 to 2016. VPS and related procedures records were reviewed. Patients between 6 months and 18 years were included. Results: A total of 285 records were reviewed, 31 samples were entered. The CSF values were, respectively, for the median and 90% percentile: total leukocytes: 0 and 7 cells/mm3, neutrophils: 0 and 6.8 cells/mm3, lymphocytes: 0 and 2 cells/mm3, proteins: 13.4 and 67.2 mg/dL, glucose: 59 and 27.4 mg/dL. Discussion: Glucose values evinced a normal rank towards the widest inferior limit with protein values exceeding the values expected. Cellularity is the variable with the lowest variation. Conclusions: The values of the CSF cytochemistry in patients with VPS are not comparable to those of the healthy population and should be interpreted according to the characteristics of this population.

Análise do líquido cérebro-espinhal em cães e gatos com afecções neurológicas / Cerebrospinal fluid analysis in dogs and cats with neurological diseases

Background: Cerebrospinal fluid (CSF) analysis is of great importance for evaluation of patients with central nervoussystem problems, because it helps to confirm or exclude many diseases when evaluated together with other relevant clinical and ancillary laboratory tests. As there are few studies in Brazil describing results of CSF analysis in dogs and cats,the objective of the present study was to evaluate results of CSF analysis in a period of ten years, grouping the changesobserved, mainly total and differential cell count and total protein concentration, according to the following etiologicalcategories of diseases: inflammatory, degenerative, traumatic, neoplastic and vascular disorders.Materials, Methods & Results: Medical records of dogs and cats with neurological diseases that had CSF collected andanalyzed at a Veterinary Teaching Hospital between 2008 and 2017 were retrospectively reviewed. Patient data includedbreed, sex, body weight, age at presentation, findings of clinical and neurological examination, results of CSF analysis,diagnostic imaging and other laboratory tests results including PCR and serologies for infectious diseases. A total of 245CSF exams were studied (240 dogs and five cats). The established normal range for protein concentration was less than25 mg/dL for cisternal collection and less than 45 mg/dL for lumbar puncture. Total nucleated cell count was considerednormal when less than 5 cells/µL, with a predominance of mononuclear cells. Both pleocytosis and increased concentrationof the protein level were classified as discrete, moderate and marked, and the pleocytosis was further classified as mononuclear, neutrophilic and mixed cell pleocytosis. The information obtained was analyzed in relation to the neurologicalsyndromes and etiological categories of neurological...

Acute phase proteins in serum and cerebrospinal fluid in healthy cattle: possible use for assessment of neurological diseases / Proteínas de fase aguda no soro e no líquido cefalorraquidiano de bovinos hígidos: possíveis usos na avaliação de doenças neurológicas

Use of acute-phase proteins (APPs) for assessment of health and disease in animals has increased greatly within the last decade. The objective was to determine the normal concentration of APPs in the serum and cerebrospinal fluid (CSF) of healthy cattle by polyacrylamide gel electrophoresis. Fifty crossbred animals (350±70kg of BW and 18±1.2 months of age), 25 heifers and 25 steers were used. CSF samples were collected from atlanto-occipital (AO) site and blood samples were obtained from the jugular vein. CSF and serum protein electrophoresis were performed by means of sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Thirty-seven proteins with molecular weights ranging from 7 and 37kDa were identified in CSF of all animals. These eight were nominally identified with immunoglobulin A and G, celuloplasmin, transferrin, albumin, α1-antitripsin, acidic glycoprotein, and haptoglobin. All protein fractions in CSF did not differ between heifers and steers. In sera, 34 proteins with molecular weights between 7 and 244kDa were identified in heifers and steers. Similar proteins were nominally identified in the sera, but only the CSF presented α1-antitripsin. The serum values of acidic glycoprotein and immunoglobulin G were significantly higher in steers compared with heifers. In conclusion, measurement of CSF acute phase protein concentrations can be useful in diagnosing and monitoring the progression of bovine neurological diseases, perhaps even to guide therapeutic procedures. The CSF electrophoretic profile of healthy cattle does not change depending on gender.(AU)

O uso de proteínas de fase aguda (PFAs) para a avaliação da saúde e da doença em animais de produção tem aumentado consideravelmente na última década. O objetivo deste estudo foi determinar a concentração normal de PAFs no soro e no líquido cefalorraquidiano (LCR) de bovinos sadios por meio da eletroforese em gel de poliacrilamida. Foram avaliados cinquenta animais mestiços (350±70kg de PV e 18±1,2 meses de idade), 25 novilhas e 25 novilhos. As amostras de LCR foram colhidas no espaço atlanto-occipital (AO) e as amostras de sangue obtidas da veia jugular. As PAFs do soro e do LCR foram determinadas através da eletroforese em gel poliacrilamida. Trinta e sete proteínas com pesos moleculares que variaram entre 7 e 37kDa foram identificadas no LCR de todos os animais, independente do sexo. Estas oito proteínas foram nominalmente identificadas como imunoglobulina A e G, ceruloplasmina, transferrina, albumina, α1-antitripsina, glicoproteína ácida, e haptoglobina. As frações de proteínas presentes no LCR não diferiram entre novilhas e novilhos. No soro de machos e fêmeas, 34 proteínas com pesos moleculares entre 7 e 244 kDa foram identificadas. As proteínas do soro foram similarmente identificadas, entretanto a α1-antitripsina foi identificada somente no LCR. Os valores séricos de glicoproteína ácida e imunoglobulina G foram significativamente mais elevados nas novilhas em comparação aos novilhos. Em conclusão, a determinação das concentrações de proteínas de fase aguda presentes do LCR pode ser útil no diagnóstico e monitoramento da progressão de doenças neurológicas bovinas, talvez possa ainda direcionar procedimentos terapêuticos. O perfil eletroforético do LCR de bovinos hígidos não se altera em função do sexo.(AU)

Acute phase proteins in serum and cerebrospinal fluid in healthy cattle: possible use for assessment of neurological diseases / Proteínas de fase aguda no soro e no líquido cefalorraquidiano de bovinos hígidos: possíveis usos na avaliação de doenças neurológicas

Use of acute-phase proteins (APPs) for assessment of health and disease in animals has increased greatly within the last decade. The objective was to determine the normal concentration of APPs in the serum and cerebrospinal fluid (CSF) of healthy cattle by polyacrylamide gel electrophoresis. Fifty crossbred animals (350±70kg of BW and 18±1.2 months of age), 25 heifers and 25 steers were used. CSF samples were collected from atlanto-occipital (AO) site and blood samples were obtained from the jugular vein. CSF and serum protein electrophoresis were performed by means of sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Thirty-seven proteins with molecular weights ranging from 7 and 37kDa were identified in CSF of all animals. These eight were nominally identified with immunoglobulin A and G, celuloplasmin, transferrin, albumin, α1-antitripsin, acidic glycoprotein, and haptoglobin. All protein fractions in CSF did not differ between heifers and steers. In sera, 34 proteins with molecular weights between 7 and 244kDa were identified in heifers and steers. Similar proteins were nominally identified in the sera, but only the CSF presented α1-antitripsin. The serum values of acidic glycoprotein and immunoglobulin G were significantly higher in steers compared with heifers. In conclusion, measurement of CSF acute phase protein concentrations can be useful in diagnosing and monitoring the progression of bovine neurological diseases, perhaps even to guide therapeutic procedures. The CSF electrophoretic profile of healthy cattle does not change depending on gender.(AU)

O uso de proteínas de fase aguda (PFAs) para a avaliação da saúde e da doença em animais de produção tem aumentado consideravelmente na última década. O objetivo deste estudo foi determinar a concentração normal de PAFs no soro e no líquido cefalorraquidiano (LCR) de bovinos sadios por meio da eletroforese em gel de poliacrilamida. Foram avaliados cinquenta animais mestiços (350±70kg de PV e 18±1,2 meses de idade), 25 novilhas e 25 novilhos. As amostras de LCR foram colhidas no espaço atlanto-occipital (AO) e as amostras de sangue obtidas da veia jugular. As PAFs do soro e do LCR foram determinadas através da eletroforese em gel poliacrilamida. Trinta e sete proteínas com pesos moleculares que variaram entre 7 e 37kDa foram identificadas no LCR de todos os animais, independente do sexo. Estas oito proteínas foram nominalmente identificadas como imunoglobulina A e G, ceruloplasmina, transferrina, albumina, α1-antitripsina, glicoproteína ácida, e haptoglobina. As frações de proteínas presentes no LCR não diferiram entre novilhas e novilhos. No soro de machos e fêmeas, 34 proteínas com pesos moleculares entre 7 e 244 kDa foram identificadas. As proteínas do soro foram similarmente identificadas, entretanto a α1-antitripsina foi identificada somente no LCR. Os valores séricos de glicoproteína ácida e imunoglobulina G foram significativamente mais elevados nas novilhas em comparação aos novilhos. Em conclusão, a determinação das concentrações de proteínas de fase aguda presentes do LCR pode ser útil no diagnóstico e monitoramento da progressão de doenças neurológicas bovinas, talvez possa ainda direcionar procedimentos terapêuticos. O perfil eletroforético do LCR de bovinos hígidos não se altera em função do sexo.(AU)

Fluid and imaging biomarkers for Alzheimer's disease: Where we stand and where to head to.

There is increasing evidence that a number of potentially informative biomarkers for Alzheimer disease (AD) can improve the accuracy of diagnosing this form of dementia, especially when used as a panel of diagnostic assays and interpreted in the context of neuroimaging and clinical data. Moreover, by combining the power of CSF biomarkers with neuroimaging techniques to visualize Aß deposits (or neurodegenerative lesions), it might be possible to better identify individuals at greatest risk for developing MCI and converting to AD. The objective of this article was to review recent progress in selected imaging and chemical biomarkers for prediction, early diagnosis and progression of AD. We present our view point of a scenario that places CSF and imaging markers on the verge of general utility based on accuracy levels that already match (or even surpass) current clinical precision.

Método de aglutinación en látex para el diagnóstico rápido del síndrome de Guillain-Barré / Latex agglutination method for fast Guillain-Barré syndrome`s diagnosis

El síndrome de Guillain-Barré es una polirradiculoneuropatía aguda, en la cual está involucrado un componente autoinmunitario, posterior a un proceso infeccioso en pacientes no vacunados o vacunados. A partir de la aparición del virus del dengue y del Zika en el continente americano es de esperar que exista una mayor probabilidad de encontrar pacientes con este síndrome post-infeccioso. Por tanto, poder contar con un método diagnóstico rápido pudiera ser de utilidad en los centros de asistencia que reciben casos de urgencias. Se procede a modificar un método para cuantificar albuminuria por aglutinación con partículas de látex sensibilizados, para la detección de este analito en el líquido cefalorraquídeo de aquellos pacientes con sospecha de esta enfermedad. Se comprueba que el método puede ser utilizado como método de diagnóstico rápido del síndrome de Guillain-Barré(AU)

Guillain-Barré Syndrome is an acute poliradiculoneuropathy with an autoimmune component, subsequent to an infectious process in vaccinated or non-vaccinated patients. From the spreading of dengue and Zika infections in the American continent it is expected a greater probability of finding patients with this post-infectious syndrome. Therefore, a rapid diagnostic test could be useful in emergency assistance centers. A quantitative latex agglutination test to detect albuminuria was modified to be used in cerebrospinal fluid of patients with presumptive diagnosis of this disease. It has been proved that the developed test can be used for diagnostic rapid of Guillain-Barré syndrome(AU)

Parâmetros citológicos e bioquímicos do líquido cefalorraquidiano coletado de bovinos sadios em dois momentos, com intervalo de 96 horas / Cerebrospinal fluid cytological and biochemical values of healthy cattle in two sampling times with an interval of 96 hours

A análise do líquido cefalorraquidiano (LCR) é uma importante ferramenta no diagnóstico das doenças neurológicas dos bovinos. A coleta do LCR em diferentes momentos facilita o monitoramento do quadro clínico e/ou a avaliação da eficácia de tratamentos estabelecidos. Todavia, os efeitos de coletas consecutivas sobre os parâmetros citológicos e bioquímicos do LCR de bovinos saudáveis ainda não foram analisados. Dessa forma, o objetivo do presente estudo foi avaliar potenciais diferenças citológicas e bioquímicas no LCR de bovinos sadios, obtidos em dois momentos, com intervalo de 96 horas. O LCR foi submetido à análise citológica e à determinação do teor de proteína total. Na segunda coleta, verificou-se aumento significativo da celularidade, porém ainda dentro do intervalo de referência para a espécie, diminuição de linfócitos e elevação de células mononucleares em relação à primeira coleta. No entanto, os números de hemácias (/µL), células nucleadas (/µL), neutrófilos (%), eosinófilos (%) e a concentração da proteína total (mg/dL) continuaram dentro dos valores de referência 96 horas após a realização da primeira punção. Conclui-se que os procedimentos de coleta de LCR consecutivos não provocaram inflamação local detectável 96 horas após a primeira punção, o que sugere que, quando necessário, coletas de LCR seriadas, por si só, não interfeririam nos resultados do exame.(AU)

The analysis of cerebrospinal fluid (CSF) is important for the diagnosis of neurological diseases in cattle. However, the effect of consecutive collections on the cytological and biochemical parameters of the CSF has not been evaluated yet. The objective of the present study was to evaluate potential differences on the cytological and biochemical analysis of CSF obtained from healthy cattle at two sampling times with 96 hours of interval in between. Total and differential cell counts, and total protein concentration were analyzed in the CSF. Increased cellularity and significant reduction of lymphocyte and elevated numbers of mononuclear cells were observed on the second collection. However, erythrocyte count (/µl), total leukocyte count (/µl), neutrophils (%), eosinophils (%) and total protein concentration (mg/dl) remained within the reference range 96 hours after the completion of the first puncture. In conclusion, consecutive CSF collections cause no detectable local inflammation 96 hours after the first puncture suggesting that repeated collections do not interfere in the CSF results.(AU)

Parâmetros citológicos e bioquímicos do líquido cefalorraquidiano coletado de bovinos sadios em dois momentos, com intervalo de 96 horas / Cerebrospinal fluid cytological and biochemical values of healthy cattle in two sampling times with an interval of 96 hours

A análise do líquido cefalorraquidiano (LCR) é uma importante ferramenta no diagnóstico das doenças neurológicas dos bovinos. A coleta do LCR em diferentes momentos facilita o monitoramento do quadro clínico e/ou a avaliação da eficácia de tratamentos estabelecidos. Todavia, os efeitos de coletas consecutivas sobre os parâmetros citológicos e bioquímicos do LCR de bovinos saudáveis ainda não foram analisados. Dessa forma, o objetivo do presente estudo foi avaliar potenciais diferenças citológicas e bioquímicas no LCR de bovinos sadios, obtidos em dois momentos, com intervalo de 96 horas. O LCR foi submetido à análise citológica e à determinação do teor de proteína total. Na segunda coleta, verificou-se aumento significativo da celularidade, porém ainda dentro do intervalo de referência para a espécie, diminuição de linfócitos e elevação de células mononucleares em relação à primeira coleta. No entanto, os números de hemácias (/µL), células nucleadas (/µL), neutrófilos (%), eosinófilos (%) e a concentração da proteína total (mg/dL) continuaram dentro dos valores de referência 96 horas após a realização da primeira punção. Conclui-se que os procedimentos de coleta de LCR consecutivos não provocaram inflamação local detectável 96 horas após a primeira punção, o que sugere que, quando necessário, coletas de LCR seriadas, por si só, não interfeririam nos resultados do exame.(AU)

The analysis of cerebrospinal fluid (CSF) is important for the diagnosis of neurological diseases in cattle. However, the effect of consecutive collections on the cytological and biochemical parameters of the CSF has not been evaluated yet. The objective of the present study was to evaluate potential differences on the cytological and biochemical analysis of CSF obtained from healthy cattle at two sampling times with 96 hours of interval in between. Total and differential cell counts, and total protein concentration were analyzed in the CSF. Increased cellularity and significant reduction of lymphocyte and elevated numbers of mononuclear cells were observed on the second collection. However, erythrocyte count (/µl), total leukocyte count (/µl), neutrophils (%), eosinophils (%) and total protein concentration (mg/dl) remained within the reference range 96 hours after the completion of the first puncture. In conclusion, consecutive CSF collections cause no detectable local inflammation 96 hours after the first puncture suggesting that repeated collections do not interfere in the CSF results.(AU)

Utilização de escore e dosagem de lactato no líquor para diagnóstico diferencial entre meningite bacteriana e meningite asséptica / Use of score and cerebrospinal fluid lactate dosage in differential diagnosis of bacterial and aseptic meningitis

RESUMO Objetivo: Avaliar o Escore para Meningite Bacteriana (EMB) isolado e associado ao valor do lactato no líquor para diferenciar meningite bacteriana (MB) e meningite asséptica (MA). Métodos: Foram selecionadas crianças com meningite atendidas em hospital terciário privado entre janeiro de 2011 e dezembro de 2014. Os dados foram obtidos na admissão. Utilizou-se o EMB com: coloração de Gram no líquor (2 pontos); neutrófilos no líquor ≥1.000 células/mm3 (1 ponto); proteína no líquor ≥80 mg/dL (1 ponto); neutrófilos no sangue periférico ≥10.000 células/mm3 (1 ponto); e convulsão durante/antes da chegada (1 ponto). Analisou-se também o lactato no líquor (elevado: ≥30 mg/dL). Avaliaram-se sensibilidade, especificidade e valor preditivo negativo de diversos valores de corte do EMB e do EMB associado ao lactato elevado para prever MB. Resultados: Dos 439 pacientes elegíveis, 94 não tinham todos os dados necessários para o escore, sendo 345 pacientes selecionados: 7 no grupo de MB e 338 no de MA. Como preditivos de MB, o EMB ≥1 mostrou sensibilidade de 100% (intervalo de confiança de 95% - IC95% 47,3-100), especificidade de 64,2% (58,8-100) e valor preditivo negativo de 100% (97,5-100), enquanto o EMB ≥2 ou EMB ≥1 associado a lactato liquórico ≥30 mg/dL mostrou sensibilidade de 100% (47,3-100), especificidade de 98,5% (96,6-99,5) e valor preditivo negativo de 100% (98,3-100). Conclusões: O EMB com 2 pontos associado à dosagem de lactato no líquor manteve a sensibilidade e o valor preditivo negativo, ao passo que aumentou a especificidade para identificar meningites bacterianas em relação à utilização do EMB com 1 ponto.

ABSTRACT Objective: To evaluate Bacterial Meningitis Score (BMS) on its own and in association with Cerebrospinal Fluid (CSF) lactate dosage in order to distinguish bacterial from aseptic meningitis. Methods: Children diagnosed with meningitis at a tertiary hospital between January/2011 and December/2014 were selected. All data were obtained upon admission. BMS was applied and included: CSF Gram staining (2 points); CSF neutrophil count ≥1,000 cells/mm3 (1 point); CSF protein ≥80 mg/dL (1 point); peripheral blood neutrophil count ≥10,000 cells/mm3 (1 point) and seizures upon/before arrival (1 point). Cutoff value for CSF lactate was ≥30 mg/dL. Sensitivity, specificity and negative predictive value of several BMS cutoffs and BMS associated with high CSF lactate were evaluated for prediction of bacterial meningitis. Results: Among 439 eligible patients, 94 did not have all data available to complete the score, and 345 patients were included: 7 in bacterial meningitis group and 338 in aseptic meningitis group. As predictive factors of bacterial meningitis, BMS ≥1 had 100% sensitivity (95%CI 47.3-100), 64.2% specificity (58.8-100) and 100% negative predictive value (97.5-100); BMS ≥2 or BMS ≥1 associated with high CSF lactate also showed 100% sensitivity (47.3-100); but 98.5% specificity (96.6-99.5) and 100% negative predictive value (98.3-100). Conclusions: 2 point BMS in association with CSF lactate dosage had the same sensitivity and negative predictive value, with increased specificity for diagnosis of bacterial meningitis when compared with 1-point BMS.

USE OF SCORE AND CEREBROSPINAL FLUID LACTATE DOSAGE IN DIFFERENTIAL DIAGNOSIS OF BACTERIAL AND ASEPTIC MENINGITIS. / UTILIZAÇÃO DE ESCORE E DOSAGEM DE LACTATO NO LÍQUOR PARA DIAGNÓSTICO DIFERENCIAL ENTRE MENINGITE BACTERIANA E MENINGITE ASSÉPTICA.

OBJECTIVE: To evaluate Bacterial Meningitis Score (BMS) on its own and in association with Cerebrospinal Fluid (CSF) lactate dosage in order to distinguish bacterial from aseptic meningitis. METHODS: Children diagnosed with meningitis at a tertiary hospital between January/2011 and December/2014 were selected. All data were obtained upon admission. BMS was applied and included: CSF Gram staining (2 points); CSF neutrophil count ≥1,000 cells/mm3 (1 point); CSF protein ≥80 mg/dL (1 point); peripheral blood neutrophil count ≥10,000 cells/mm3 (1 point) and seizures upon/before arrival (1 point). Cutoff value for CSF lactate was ≥30 mg/dL. Sensitivity, specificity and negative predictive value of several BMS cutoffs and BMS associated with high CSF lactate were evaluated for prediction of bacterial meningitis. RESULTS: Among 439 eligible patients, 94 did not have all data available to complete the score, and 345 patients were included: 7 in bacterial meningitis group and 338 in aseptic meningitis group. As predictive factors of bacterial meningitis, BMS ≥1 had 100% sensitivity (95%CI 47.3-100), 64.2% specificity (58.8-100) and 100% negative predictive value (97.5-100); BMS ≥2 or BMS ≥1 associated with high CSF lactate also showed 100% sensitivity (47.3-100); but 98.5% specificity (96.6-99.5) and 100% negative predictive value (98.3-100). CONCLUSIONS: 2 point BMS in association with CSF lactate dosage had the same sensitivity and negative predictive value, with increased specificity for diagnosis of bacterial meningitis when compared with 1-point BMS.

OBJETIVO: Avaliar o Escore para Meningite Bacteriana (EMB) isolado e associado ao valor do lactato no líquor para diferenciar meningite bacteriana (MB) e meningite asséptica (MA). MÉTODOS: Foram selecionadas crianças com meningite atendidas em hospital terciário privado entre janeiro de 2011 e dezembro de 2014. Os dados foram obtidos na admissão. Utilizou-se o EMB com: coloração de Gram no líquor (2 pontos); neutrófilos no líquor ≥1.000 células/mm3 (1 ponto); proteína no líquor ≥80 mg/dL (1 ponto); neutrófilos no sangue periférico ≥10.000 células/mm3 (1 ponto); e convulsão durante/antes da chegada (1 ponto). Analisou-se também o lactato no líquor (elevado: ≥30 mg/dL). Avaliaram-se sensibilidade, especificidade e valor preditivo negativo de diversos valores de corte do EMB e do EMB associado ao lactato elevado para prever MB. RESULTADOS: Dos 439 pacientes elegíveis, 94 não tinham todos os dados necessários para o escore, sendo 345 pacientes selecionados: 7 no grupo de MB e 338 no de MA. Como preditivos de MB, o EMB ≥1 mostrou sensibilidade de 100% (intervalo de confiança de 95% - IC95% 47,3-100), especificidade de 64,2% (58,8-100) e valor preditivo negativo de 100% (97,5-100), enquanto o EMB ≥2 ou EMB ≥1 associado a lactato liquórico ≥30 mg/dL mostrou sensibilidade de 100% (47,3-100), especificidade de 98,5% (96,6-99,5) e valor preditivo negativo de 100% (98,3-100). CONCLUSÕES: O EMB com 2 pontos associado à dosagem de lactato no líquor manteve a sensibilidade e o valor preditivo negativo, ao passo que aumentou a especificidade para identificar meningites bacterianas em relação à utilização do EMB com 1 ponto.

Polyspecific antibodies without persisting antigen in multiple sclerosis, neurolupus and Guillain-Barré syndrome: immune network connectivity in chronic diseases.

The polyspecific antibody synthesis in multiple sclerosis (MS) gained diagnostic relevance with the frequent combination of measles-, rubella- and varicella zoster antibodies (MRZ-antibody reaction) but their pathophysiological role remains unknown. This review connects the data for intrathecal polyspecific antibody synthesis in MS and neurolupus with observations in the blood of patients with Guillain-Barré syndrome (GBS). Simultaneously increased antibody and autoantibody titers in GBS blood samples indicate that the polyspecific antibodies are based on a general property of an immune network, supported by the deterministic day-to-day concentration variation of antibodies in normal blood. Strongly correlated measles- and rubella- antibody variations point to a particular connectivity between the MRZ antibodies. The immune network, which provides serological memory in the absence of an antigen, implements the continuous change of the MRZ pattern in blood, not followed by the earlier immigrated B cells without corresponding connectivity in the brain. This may explain the different antibody patterns in cerebrospinal fluid, aqueous humor and blood of the individual MS patient. A complexity approach must implement a different view on causation in chronic diseases and causal therapies.

Polyspecific antibodies without persisting antigen in multiple sclerosis, neurolupus and Guillain-Barré syndrome: immune network connectivity in chronic diseases / Anticorpos poliespecíficos sem antígeno persistente em esclerose múltipla, neurolupus e síndrome de Guillain-Barré: conectividade imunológica em doenças crônicas

ABSTRACT The polyspecific antibody synthesis in multiple sclerosis (MS) gained diagnostic relevance with the frequent combination of measles-, rubella- and varicella zoster antibodies (MRZ-antibody reaction) but their pathophysiological role remains unknown. This review connects the data for intrathecal polyspecific antibody synthesis in MS and neurolupus with observations in the blood of patients with Guillain-Barré syndrome (GBS). Simultaneously increased antibody and autoantibody titers in GBS blood samples indicate that the polyspecific antibodies are based on a general property of an immune network, supported by the deterministic day-to-day concentration variation of antibodies in normal blood. Strongly correlated measles- and rubella- antibody variations point to a particular connectivity between the MRZ antibodies. The immune network, which provides serological memory in the absence of an antigen, implements the continuous change of the MRZ pattern in blood, not followed by the earlier immigrated B cells without corresponding connectivity in the brain. This may explain the different antibody patterns in cerebrospinal fluid, aqueous humor and blood of the individual MS patient. A complexity approach must implement a different view on causation in chronic diseases and causal therapies.

RESUMO A síntese de anticorpos poliespecíficos em esclerose múltipla (EM) ganhou relevância diagnóstica com a combinação frequente de anticorpos contra sarampo, rubéola e varicela-zoster (reação de anticorpos MRZ), mas seu papel fisiopatológico permanece desconhecido. Esta revisão relaciona os dados da síntese intratecal de anticorpos poliespecíficos em EM e Neurolupus com observações no sangue de pacientes com síndrome de Guillain Barré (SGB). Simultaneamente, os títulos aumentados de anticorpos e autoanticorpos em amostras de sangue de SGB indicam que os anticorpos poliespecíficos se baseiam numa propriedade geral de uma rede imunitária, suportada pela variação determinística da concentração diária de anticorpos no sangue normal. As variações fortemente correlacionadas de anticorpos contra sarampo e rubéola apontam para uma conectividade particular entre os anticorpos MRZ. A rede imunitária, que fornece memória sorológica na ausência de um antígeno, implementa a mudança contínua do padrão MRZ no sangue, não seguida pelas células B que imigraram anteriormente sem conectividade no cérebro. Isto pode explicar os diferentes padrões de anticorpos no LCR, humor aquoso e sangue do paciente individual de EM. Uma abordagem complexa deve implementar uma visão diferente sobre a causalidade em doenças crônicas e terapias causais.

Association between Clinical Outcomes and Hospital Guidelines for Cerebrospinal Fluid Testing in Febrile Infants Aged 29-56 Days.

OBJECTIVE: To describe the association between clinical outcomes and clinical practice guidelines (CPGs) recommending universal cerebrospinal fluid (CSF) testing in the emergency department for febrile infants aged 29-56 days. STUDY DESIGN: Using 2007-2013 administrative data from 32 US children's hospitals, we performed a difference-in-differences analysis comparing 7 hospitals with CPGs recommending universal CSF testing for older febrile infants aged 29-56 days (CPG group) with 25 hospitals without such CPGs (control group). We compared differences in clinical outcomes between older febrile infants with the corresponding differences among younger febrile infants aged 7-28 days. The primary outcome was the occurrence of an adverse event, defined as a delayed diagnosis of bacterial meningitis, mechanical ventilation, placement of a central venous catheter, extracorporeal membrane oxygenation, or in-hospital mortality. Analyses were adjusted for race/ethnicity, sex, median annual household income by zip code, primary insurance source, discharge season, and discharge year. RESULTS: The proportion of older febrile infants undergoing CSF testing was higher (P < .001) in the CPG group (64.8%) than the control group (47.8%). CPGs recommending universal CSF testing for older febrile infants were not associated with significant differences in adverse events (difference-in-differences: +0.31 percentage points, 95% CI -0.18 to 0.85; P = .22). CONCLUSIONS: Hospital CPGs recommending universal CSF testing for febrile infants aged 29-56 days were not associated with significant differences in clinical outcomes.

Chemical and cytological analysis of cerebral spinal fluid after intrathecal injection of hypodense fluorescein / Análise química e citológica do líquor após injeção intratecal de solução hipodensa de fluoresceína

ABSTRACT INTRODUCTION: Intrathecal fluorescein has been effective for topographic diagnosis of rhinoliquorrhea. Nonetheless, there are no reports on the study of cerebral spinal fluid (CSF) after use of intrathecal fluorescein. OBJECTIVE: A prospective study attempting to evaluate CSF through chemical and cytological analysis, after injection of fluorescein. METHODS: Prospective analysis of 24 samples of CSF after intrathecal injection of fluorescein for topographic diagnosis of CSF fistulae, collected at the time of puncture and after 24 and 48 h, divided by cellularity: Group 1, up to five cells, and Group 2, with more than five cells. RESULTS: The yellow-greenish color of CSF remained after 48 h in 36%, evidencing permanence of fluorescein. No changes in protein and glucose levels were observed between 0-24 h and 0-48 h. In group 2, an increase in cell count was observed between 24 h and 48 h (p = 0.019). In both groups, there was an increase of neutrophils between 0 and 48 h (p = 0.048) and a decrease between 24 and 48 h (p = 0.05). CONCLUSION: Intrathecal fluorescein provoked discreet meningeal reactions, such as an increase of cells between 24 and 48 h and an increase of neutrophils at 24 h, with a subsequent decrease at 48 h with no correlation with symptomatology.

RESUMO Introdução: A fluoresceína intratecal tem sido efetiva no diagnóstico topográfico da rinoliquorréia. Entretanto, não há estudos no líquor após o uso de fluoresceína intratecal. Objetivo: Estudo prospectivo visando avaliar o líquor, através de análise química e citológica, após injeção de fluoresceína. Método: Análise prospectiva de 24 punções após injeção intratecal de fluoresceína para diagnóstico topográfico de fístula liquórica, coletado no momento da punção, 24 e 48 horas, divididos pela celularidade: grupo 1, com até 5 células e grupo 2 com mais de 5 células. Resultado: A coloração amarelo-esverdeada do líquor permaneceu após 48 horas em 36%, evidenciando permanência de fluoresceína. Observou-se ausência de mudanças no nível de proteína e glicose entre 0-24 horas e 0-48 horas. No grupo 2, um aumento na contagem celular foi observado entre 24 e 48 horas (p = 0,019). No dois grupos juntos, observou-se um aumento de neutrófilos entre 0 e 48 horas (p = 0,048) e uma diminuição entre 24 e 28 horas (p = 0,05). Conclusão: Fluoresceína intratecal provocou discretas reações meníngeas, como o aumento de células entre 24 e 48 horas e aumento dos dos neutrófilos em 24 horas com uma subsequente dimi nuição em 48 horas sem correlação com sintomas.

Chemical and cytological analysis of cerebral spinal fluid after intrathecal injection of hypodense fluorescein.

INTRODUCTION: Intrathecal fluorescein has been effective for topographic diagnosis of rhinoliquorrhea. Nonetheless, there are no reports on the study of cerebral spinal fluid (CSF) after use of intrathecal fluorescein. OBJECTIVE: A prospective study attempting to evaluate CSF through chemical and cytological analysis, after injection of fluorescein. METHODS: Prospective analysis of 24 samples of CSF after intrathecal injection of fluorescein for topographic diagnosis of CSF fistulae, collected at the time of puncture and after 24 and 48h, divided by cellularity: Group 1, up to five cells, and Group 2, with more than five cells. RESULTS: The yellow-greenish color of CSF remained after 48h in 36%, evidencing permanence of fluorescein. No changes in protein and glucose levels were observed between 0-24h and 0-48h. In group 2, an increase in cell count was observed between 24h and 48h (p=0.019). In both groups, there was an increase of neutrophils between 0 and 48h (p=0.048) and a decrease between 24 and 48h (p=0.05). CONCLUSION: Intrathecal fluorescein provoked discreet meningeal reactions, such as an increase of cells between 24 and 48h and an increase of neutrophils at 24h, with a subsequent decrease at 48h with no correlation with symptomatology.