Determination of sulfur compounds in biological desulfurization system by high performance liquid chromatography / 生物工程学报

Biological desulfurization is a process in which sulfur compounds are removed from gas and oil using microorganisms. It is a simple process that has mild operating conditions, high desulfurization efficiency, low energy consumption and less environmental pollution. However, there is still a lack of simple and efficient analytical methods for quantitatively analyzing the sulfur compounds in the biological desulfurization process. In order to solve this problem, the analytical method for the simultaneous determination of sulfite, thiosulfate and sulfide in biological desulfurization solutions by pre-column fluorescence derivation using high performance liquid chromatography (HPLC) was developed. The standard curves of sulfur species in this analytical method had good linear relationships with correlation coefficients of 0.999 5, 0.999 7, and 0.999 7 for sulfite, thiosulfate and sulfide, respectively. The detection limits of these sulfur compounds were 0.000 6, 0.000 7 and 0.001 1 μmol/L; the range of recovery rates were 98.17 to 101.9%, 100.9 to 102.6%, and 101.1 to 104.2%; which had good repeatability and stability. The analytical method was simple, efficient and accurate, and could be used to simultaneously determine the sulfur compounds in different biological desulfurization systems.

Mass Spectral Character of Fentanyl Analogues / 法医学杂志

Objective To provide the reference for the identification of unknown fentanyl analogues by studying the characteristic ions and main fragmentation pathways of fentanyl analogues in the modes of collision induced dissociation （CID） and electron ionization （EI）. Methods Nine fentanyl analogues （2, 2'-difluorofentanyl, acetyl fentanyl, fentanyl, butyl fentanyl, valeryl fentanyl, acryloyl fentanyl, furan fentanyl, 4-fluorine isobutyl fentanyl, carfentanyl） were selected and analyzed with ultra-high performance liquid chromatography-quadrupole time-of-flight-mass spectrometry （UHPLC-QTOF-MS） and gas chromatography-mass spectrometry （GC-MS）. The mass spectrum obtained was analyzed. The CID and EI fragmentation routes of fentanyl analogues were speculated. Results The CID and EI fragmentation pathways were highly similar. In the CID mode, characteristic ions were formed by the carbon-nitrogen bond cleavage between the piperidine ring and the N-phenyl-amide moiety, within the piperidine ring, and between the phenethyl and piperidine ring. While in the EI mode, dissociation of the piperidine ring, as well as cleavage between the piperidine ring and the phenethyl were the main fragmentation pathways. Conclusion This study summarizes the main fragmentation pathways and characteristic ions of fentanyl analogues in the CID and EI modes, which is useful for forensic laboratories to identify and structural analyze fentanyl type new psychoactive substance in practical work.

Seno preauricular, diferentes tecnicas quirúrgicas / Preauriculares Sinus, different surgical techniques

El seno preauricular es una malformación congénita que se presenta como un pequeño orificio en el oído externo, usualmente cercano al borde anterior de la rama ascendente del hélix. La mayoría son asintomáticos y no requieren tratamiento, sin embargo, una vez infectados se vuelven inflamaciones dolorosas, con secreción fétida que presentan exacerbaciones agudas recurrentes. Objetivo: brindar información actualizada sobre las ventajas de las diferentes técnicas quirúrgicas utilizadas para resolver en forma adecuada este problema congénito. Material y Métodos : el presente estudio se realizó mediante una búsqueda comprensiva de artículos con menos de 10 años de publicación en las bases de datos de MEDLINE/PubMed, Google Académico e HINARI. Se seleccionaron un total de 20 artículos, en español e inglés, entre ellos trabajos originales y metanálisis sobre el tema. Conclusión : la técnica suprauricular ha demostrado, en términos estadísticos, una tasa de recurrencia menor que la técnica clásica de sinectomía...(AU)

Evaluation of methods for the quantification of ether extract contents in forage and cattle feces

ABSTRACT The objective of this study was to compare the estimates of ether extract (EE) contents obtained by the Randall method and by the high-temperature method of the American Oil Chemist's Society (AOCS; Am 5-04) in forages (n = 20) and cattle feces (n = 15). The EE contents were quantified by using the Randall extraction or AOCS method and XT4 filter bags or cartridges made of qualitative filter paper (80 g/m²) as containers for the samples. It was also evaluated the loss of particles, and concentration of residual chlorophyll after extraction and the recovery of protein and minerals in the material subjected to extraction. Significant interaction was observed between extraction method and material for EE contents. The EE estimates using the AOCS method were higher, mainly in forages. No loss of particles was observed with different containers. The chlorophyll contents in the residues of cattle feces were not affected by the extraction method; however, residual chlorophyll was lower using the AOCS method in forages. There was complete recovery of the protein and ash after extraction. The results suggest that AOCS method produces higher estimates of EE contents in forages and cattle feces, possibly by providing greater extraction of non-fatty EE.

Antioxidant activity of bioactive extracts obtained from rhizomes of Cyperus digitatus Roxb / Actividad antioxidante de extractos obtenidos a partir de rizomas de Cyperus digitatus Roxb

Members of the family Cyperaceae such as Cyperus alopecuroides, Cyperus articulatus, Cyperus scariosus and Cyperus rotundus possess significant amount of studies about their antioxidant activities and other properties. Nevertheless, the plant Cyperus digitatus belonging to the genus Cyperus lacks of studied about any kind of intrinsic activity. Different extracts and fractions were obtained from the rhizomes of Cyperus digitatus, and a Phytochemical screening and the content of phenols and flavonoids and the antioxidant properties (FRAP, DPPH and beta-Carotene bleaching) were quantified in each of theextracts and fractions. Of all the extracts obtained, the BE and AqE extracts showed the best antioxidant potential, meanwhile, none of the fractions obtained from the EAE extract show a relevant activity.

Los miembros de la familia Cyperaceae, tales como Cyperus alopecuroides, Cyperus articulatus, Cyperus scariosus y Cyperus rotundus poseen una cantidad significativa de estudios sobre sus actividades antioxidantes y otras propiedades. Sin embargo, la planta Cyperus digitatus perteneciente al género Cyperus carece de estudio de cualquier tipo de actividad intrínseca. Razón por la cual se estudió sus propiedades antioxidantes (FRAP, DPPH y blanqueamiento del beta-caroteno), cuantificación de contenido fenolico y flavonoides totales en extractos y fracciones obtenidos de los rizomas de Cyperus digitatus, y un perfil fitoquímico. De todos los extractos obtenidos, BE y AqE mostraron el mejor potencial antioxidante, por otra parte ninguna de las fracciones obtenidas a partir del extracto EAE mostro una actividad relevante.

Development of a monoclonal antibody against deoxynivalenol for magnetic nanoparticle-based extraction and an enzyme-linked immunosorbent assay

Monoclonal antibody (mAb, NVRQS-DON) against deoxynivalenol (DON) was prepared. DON-Ag coated enzyme linked immunosorbent assay (ELISA) and DON-Ab coated ELISA were prepared by coating the DON-BSA and DON mAb. Quantitative DON calculation ranged from 50 to 4,000 ng/mL for DON-Ab coated ELISA and from 25 to 500 ng/mL for DON-Ag coated ELISA. 50% of inhibitory concentration values of DON, HT-2, 15-acetyl-DON, and nivalenol were 23.44, 22,545, 5,518 and 5,976 ng/mL based on the DON-Ab coated ELISA. Cross-reactivity levels of the mAb to HT-2, 15-acetyl-DON, and nivalenol were 0.1, 0.42, and 0.40%. The intra- and interassay precision coefficient variation (CV) were both <10%. In the mAb-coated ELISA, mean DON recovery rates in animal feed (0 to 1,000 microg/kg) ranged from 68.34 to 95.49% (CV; 4.10 to 13.38%). DON in a buffer solution (250, 500 and 1,000 ng/mL) was isolated using 300 microg of NVRQS-DON and 3 mg of magnetic nanoparticles (MNPs). The mean recovery rates of DON using this mAb-MNP system were 75.2, 96.9, and 88.1% in a buffer solution spiked with DON (250, 500, and 1,000 ng/mL). Conclusively we developed competitive ELISAs for detecting DON in animal feed and created a new tool for DON extraction using mAb-coupled MNPs.

Nanochips y nanosensores para eldiagnóstico temprano de cáncer oral: una revisión / Nanochips and nanosensors for harly oral cancer diagnosis: a review

Antecedentes: El cáncer oral es una de las enfermedades más agresivas y con mayor probabilidadde metástasis; en estadios iniciales es generalmente indetectable y de baja consulta,por lo que se dificulta realizar un tratamiento conservador. Existen diversas técnicas delaboratorio con base en microscopía (como inmunofluorescencia), inmunohistoquímica yotras que facilitan el diagnóstico temprano de la enfermedad. Adicionalmente, los avancesen nanomedicina brindan nuevas herramientas de detección a partir de cambios moleculares(biomarcadores) que presentan las células en proceso de malignización. Objetivo:Describir las características clínicas y moleculares de lesiones premalignas y cáncer oral ylos métodos de diagnóstico usando nanotecnología (nanochips, nanosensores, etc.), comoun método eficaz para la detección temprana del cáncer. Método: Se buscó literatura sobrenanotecnología y diagnóstico de cáncer oral en bases de datos como Science Direct yPubMed; la información de cada uno de los artículos se sintetizó con resúmenes individualespor los investigadores, se agrupó y redactó. Resultados: Se seleccionaron 46 artículos que,de acuerdo con su contenido, se agruparon según su temática principal; toda la informaciónse relacionó en una tabla por subtemas en Excel® 2007. Conclusiones: La literatura revisadasugiere que las herramientas nanotecnológicas pueden ser una alternativa útil y rápida,aunque por el momento costosa, para la detección puntual de biomarcadores presentes enestadios iniciales del cáncer oral. Cabe mencionar que su aplicación clínica en países comoColombia es limitada por factores como insuficientes recursos asignados a la investigacióny poca investigación en esta temática...

Background: Oral cancer is one of the most aggressive diseases with a high likelihood ofmetastases; it is undetectable during initial stages with low medical visits, making it difficultto provide conservative treatment. Several techniques such as microscopy (using for instanceimmunofluorescence) and immunohistochemistry make early diagnosis and cancerdetection easier. In addition, advances in nanomedicine offer new tools to detect molecularchanges (biological markers/biomarkers) in cells during a malignant process. Purpose:Describe molecular and clinic characteristics of premalignant lesions and oral cancer, aswell as techniques involving nanotechnology (lab-on-a-chip devices, nanosensors, etc.) aseffective methods for early cancer diagnosis. Methods: A literature review on nanotechnologyand oral cancer using databases such as Science Direct and PubMed was carried out;data from each article were summarized, grouped, and described. Results: 46 articles wereselected and grouped according to their main topic; data were compiled in an Excel 2007spreadsheet. Conclusions: The literature reviewed suggests that tools using nanotechnologycan be a useful and a quick alternative, though expensive, to detect specific biomarkers thatare present in early stages of oral cancer. It is important to point out that their clinical usein countries like Colombia is still limited by factors like the lack of resources allocated forresearch and the little research carried out on this subject...

Caracterização analítica de sucos e néctares de laranja adoçados com sacarose e edulcorantes / Analytical characterization of orange juices and nectars sweetened with sucrose and sweet flavoring agents

Objetivo: Caracterizar físico-quimicamente, sucos não adoçados e néctares de laranja adoçados com sacarose ou edulcorantes, quanto ao seu pH, acidez titulável (AT) e teor de sólidos solúveis totais (SST) e avaliar a correlação desta última propriedade com as demais. Método: Analisaram-se alíquotas de três lotes de dois sucos de laranja e de dois néctares com adição de sacarose ou dois com edulcorantes. Água mineral foi empregada como controle. O teor de SST foi determinado em refratômetro de Abbé. O pH foi registrado em peagômetro digital, enquanto a AT foi quantificada titulando-se amostras das bebidas com NaOH 0,1 M até o alcance dos pHs 5,5 e 7,0. Os dados foram submetidos ao teste de correlação de Pearson, análise de regressão, análise de variância e teste de Tukey (alfa = 0,05). Resultados: Os teores de SST apresentaram forte correlação com a AT, sendo a relação entre elas do tipo quadrática. Embora os valores de pH não sejam dependentes da presença de sacarose ou edulcorantes, se as bebidas são isentas dos mesmos, significativamente maior quantidade de base foi necessária para que se atingissem os pHs 5,5 e 7,0. Conclusão: Sucos e néctares de laranja apresentaram valores semelhantes de pH, os quais não se correlacionaram com a presença de sacarose ou edulcorantes nas bebidas. A acidez titulável foi maior para o suco e menor para os néctares, independentemente do fato de possuírem sacarose ou edulcorantes em sua composição. A elevação do teor de sólidos solúveis totais não implicou em redução da acidez titulável das bebidas.

Objective: To characterize physically and chemically, non-sweetened orange juices and orange nectars sweetened with sucrose or sweet flavoring agents, with respect to their pH, titratable acidity (TA) and total soluble solids content (TSSC), as well as to evaluate the correlation of the latter property with the others. Method: Aliquots of three lots of two orange juices and two orange nectars containing sucrose and two containing sweet flavoring agents were evaluated. Mineral water was used as a control. The TSSC was determined using an Abbe refractometer. The pH was recorded using a digital pH meter, while TA was quantified by titrating samples of the beverages with 0.1 M NaOH until reaching pHs 5.5 and 7.0. Data were subjected to Pearson's correlation test, regression analysis, analysis of variance and Tukey's test (alpha=0.05). Results: TSSC values presented a strong correlation with TA, and these properties exhibited a quadratic relationship. Although the pH values were not dependent on the presence of sucrose or sweet flavoring agents, a significantly greater amount of base was necessary to reach pHs 5.5 and 7.0 in the beverages without sucrose or flavoring agents. Conclusion: Orange juices and nectars presented similar pH values, which was not associated with the presence of sucrose or sweet flavoring agents in the beverages. Higher TA values were obtained for the juice and lower for the nectars, regardless of containing sucrose or sweet flavoring agents. The increase of TSSC did not implicate in decrease of TA in the beverages.

Impacto de la medición de creatinina en la estimación de la velocidad de filtración glomerular / Impact of creatinine measurement on glomerular filtration rate estimation / Impacto da medição de creatinina na estimativa da velocidade de filtração glomerular

Con el objeto de detectar precozmente la enfermedad renal crónica, muchas sociedades científicas han recomendado incorporar a los informes de laboratorio la velocidad de filtración glomerular (VFG) estimada por fórmulas asociadas a creatinina plasmática (Cr) como marcador de función renal. Frente a la variedad de metodologías disponibles en Argentina, los bioquímicos se encuentran frente al dilema de la elección del método a utilizar para cuantificar Cr. El objetivo de este trabajo es analizar si la VFG estimada por fórmulas a partir de Cr dosadas por diferentes métodos son comparables. Si bien no se observaron diferencias estadísticamente significativas entre las Cr cuantificadas por los métodos Jaffé cinético (JC), Jaffé cinético con compensación (JCCC) y enzimático (ENZ), no se obtuvo una correlación adecuada entre los mismos. Se observó que el método de Jaffé cinético sin compensación arrojó resultados de creatinina sérica, que al trasladarse a ecuaciones que determinan la VFG, dejaron un margen de error inaceptable (circunstancia que no se obser­vó de igual forma en el JCCC). Se concluye que la VFG estimada por fórmulas a partir de Cr dosadas por los métodos JC y JCCC, tomando como referencia el método enzimático, no son comparables.

In order to do early detection of chronic renal disease, many scientific societies have recommended to report a GFR estimate usinga prediction equation associated with serum creatinine measurement, as a marker of renal function. Faced with the wide variety of available creatinine measurement, as a marker of renal function. Faced with the wide variety of available creatinine methodologies in this country, professionals have a dilemma: the choice of which method must be used to quantify creatinine. The purpose of this study is to analyze whether the estimates of GFR usingcreatinine measures by different methods are comparable. Although no statistically significant differences between Cr quantified by Kinetic Jaffe method (JC), compensated kinetic Jaffe (JCCC) and enzymatic (ENZ) were observed, no good correlation was obtained between them. It was observed that the kinetic Jaffe method without compensation showed results that, beingincorporated in equations that determine the GFR, left an unacceptable margin of error (a circumstance that was not observed in the JCCC). Conclusion: GFRestimated with formulas usingcreatinine measured by JC and JCCC compared to ENZ are not comparable.

Visando detectar precocemente a doençã renal crônica, muitas sociedades científicas têm recomendado incorporar aos relatórios de laboratório a velocidade de filtração glomerular (VFG) estimada por fórmulas associadas a creatinina plasmática (Cr) como marcador de função renal. Frente à variedade de metodologias disponíveis na Argentina, os bioquímicos se encontram frente ao dilema da seleção do método a utilizar para quantificar Cr. O objetivo deste trabalho é analisar se a VFGestimada por fórmulas a partir de Cr dosadas por diferentes métodos são comparáveis. Embora não se observassem diferenças estatisticamente significativas entre as Cr quantificadas pelos métodos Jaffé cinético (JC), Jaffé cinético com compensação (JCCC) e enzimático (ENZ), não foi obtida uma correlação adequada entre os mesmos. Observouse que o método de Jaffe cinético sem compensação deuresultados de creatinina sérica, que ao se trasladarem para equações que determinam a VFG, deixaram uma margem de erro inaceitável (circunstância que não se observoude igual forma no JCCC). Conclusão: a VFGestimada por fórmulas a partir de Cr dosadas pelos métodos JC e JCCC, tomando como referência o método enzimático, não são comparáveis.

Intervalos de referencia de determinaciones bioquímicas en el laboratorio central del Hospital de Trelew / Reference intervals of biochemical determinations in the central laboratory in Trelew Hospital / Intervalos de referencia de determinagóes bioquímicas no laboratorio central do Hospital de Trelew

El objetivo del presente trabajo fue establecer los intervalos de referencia de las determinaciones: glucosa, urea, colesterol, proteínas totales, albúmina, ácido úrico, creatinina, hematocrito y hemoglobina en el Laboratorio Central del Hospital Zonal de Trelew. La población bajo estudio fueron pacientes mayores de 18 años atendidos por consultorio externo entre diciembre de 2008 y marzo de 2009. El estudio fue completado entre marzo y abril de 2011. Las determinaciones se realizaron con un autoanalizador de química clínica Metrolab 2300 plus y un contador hematológico Sysmex 2100. Se comprobó que los valores de las determinaciones se ajustaran a una distribución normal y se realizó el cálculo de los fractiles 0,025 y 0,975 para la obtención del intervalo de referencia (IR) del 95%. En un caso se utilizó transformación logarítmica de los datos y para dos categorías se aplicó el método no paramétrico. Para los valores de referencia inferior y superior se establecieron los intervalos con un 90% de confianza (IC). Los valores de referencia obtenidos fueron: glucosa de 0,74 a 1,07 g/L, urea de 0,19 a 0,51 g/L, colesterol de 1,21 a 2,43 g/L, proteínas totales de 6,45 a 7,99 g/dL, albúmina de 3,62 a 4,61 g/dL, creatinina de 0,57 a 1,10 mg/dL, ácido úrico para el sexo femenino de 18,69 a 51,93 mg/L y para el sexo masculino de 30,50 a 62,92 mg/L, hematocrito para sexo femenino de 37 a 45% y para el sexo masculino de 40 a 50%, hemoglobina para el sexo femenino de 11,70 a 15,17 g/dL y para el sexo masculino de 13,09 a 17,19 g/dL. Los valores de ácido úrico, hematocrito y hemoglobina se separaron por sexo para dar continuidad a la política del laboratorio y de los fabricantes de CAICYTtivos, que consiste en considerar las diferencias existentes entre ambos sexos. Dentro del rango dado por el fabricante se obtuvieron los resultados para glucosa, albúmina y hemoglobina; sobre el mismo, para urea, colesterol, proteínas y ácido úrico y por debajo del mismo para creatinina.

The aim of the present work was to establish the reference intervals of the following determinations, glucose, urea, cholesterol, total proteins, albumin, uric acid, creatinine, hematocrite and hemoglobin in the Central Laboratory of Trelew Zonal Hospital. The population under study was defined as 18-year-old or older patients that came to the laboratory from the ambulatory consulting room since December 2008 to March 2009, and then between March and April 2011. Blood samples were processed with a Metrolab 2300 plus clinical chemistry autoanalyser and a Sysmex 2100 hematological counter. After checking if the result distribution applied to a normal distribution, fractiles 0.025 and 0.975 were calculated to obtain a 95% Reference Interval (RI). In one case, a logarithmic transformation of the results was needed and for two categories a non-parametric method was used. For the upper and lower reference values, the intervals were calculated at 90% confidence (CI) The reference values obtained were; 0.74-1.07 g/L glucose, 0.19-0.51 g/L urea, 1.21-2.43 g/L cholesterol, 6.45-7.99g/dL total proteins, 3.62-4.61 g/dL albumin and 0.57-1.10 mg/dL creatinine, 18.69 - 51.93 mg/L uric acid in women, and 30.50 - 69.92 mg/L in men; 37 - 45% hematocrite in women and 40 - 50% in men; 11.70 - 15.17 g/dL hemoglobin in women and 13.09 - 17.19 g/dL hemoglobin in men. Uric acid, hematocrite and hemoglobin values were calculated according to sex in order to offer concordance with commercial kit and the laboratory policies which consist in considering the significant differences between both sexes. The results obtained for glucose, albumin and hemoglobin were within the reference interval given by the commercial kits; urea, cholesterol, total protein and uric acid were above it; and creatinine reference interval was lower than the reference interval from the commercial kits.

O objetivo do presente trabalho foi estabelecer os intervalos de referencia das determinagóes: glicose, ureia, colesterol, proteínas totais, albumina, ácido úrico, creatinina, hematocrito e hemoglobina no Laboratorio Central do Hospital Zonal de Trelew. A populagao sob estudo foram pacientes de mais de 18 anos atendidos através de consultorio externo entre dezembro do ano 2008 e margo de 2009. O estudo foi completado entre margo e abril de 2011. As determinagóes foram realizadas com um auto-analisador de química clínica Metrolab 2300 plus e um contador hematológico Sysmex 2100. Comprovouse que os valores das determinagóes se ajustaram a uma distribuigao normal e se realizou o cálculo dos quantis 0,025 e 0,975 para a obtengao do intervalo de referencia (IR) de 95%. Num caso foi utilizada transformagao logarítmica dos dados e para duas categorias se aplicou o método nao paramétrico. Para os valores de referencia inferior e superior foram estabelecidos os intervalos com 90% de confianga (IC). Os valores de referencia obtidos foram: glicose de 0,74 a 1,07 gr/L, ureia de 0,19 a 0,51 gr/L, colesterol de 1,21 a 2,43 g/L, proteínas totais de 6,45 a 7,99 gr/dL, albumina de 3,62 a 4,61 gr/dL, creatinina de 0,57 a 1,10 mg/dL, ácido úrico para o sexo feminino de 18,69 a 51,93 mg/L e para o sexo masculino de 30,50 a 62,92 mg/l, hematocrito para sexo feminino de 37 a 45% e para o sexo masculino de 40 a 50%, hemoglobina para o sexo feminino de 11,70 a 15,17 g/dL e para o sexo masculino de 13,09 a 17,19 g/dL. Os valores de ácido úrico, hematocrito e hemoglobina foram separados por sexo para dar continuidade a política do laboratorio e dos fabricantes de reagentes, que consiste em considerar as diferengas existentes entre ambos os sexos. Dentro do intervalo dado pelo fabricante foram obtidos os resultados para glicose, albumina e hemoglobina; superior ao mesmo para ureia, colesterol, proteínas e ácido úrico e inferior ao mesmo para creatinina.

Alternative method for quantification of alfa-amylase activity / Alternativa para quantificação de alfa-amilase

A modification of the sensitive agar diffusion method was developed for macro-scale determination of alfa-amylase. The proposed modifications lower costs with the utilisation of starch as substrate and agar as supporting medium. Thus, a standard curve was built using alfa-amylase solution from Aspergillus oryzae, with concentrations ranging from 2.4 to 7,500 U.mL-1. Clear radial diffusion zones were measured after 4 hours of incubation at 20 °C. A linear relationship between the logarithm of enzyme activities and the area of clear zones was obtained. The method was validated by testing α-amylase from barley at the concentrations of 2.4; 60; 300 and 1,500 U.mL-1. The proposed method turned out to be simpler, faster, less expensive and able to determine on a macro-scale α-amylase over a wide range (2.4 to 7,500 U.mL-1) in scientific investigation as well as in teaching laboratory activities.

Modificações foram propostas ao método sensível de difusão em ágar para a macrodeterminação de alfa-amilase. As modificações propostas diminuem os custos, com a utilização de amido como substrato e ágar como meio solidificante. Assim, foi construída uma curva padrão utilizando uma solução de alfa-amilase de Aspergillus oryzae com concentrações variando de 2,4 a 7.500 U.mL-1. Em seguida, as zonas claras de difusão radial foram mensuradas depois de 4 horas de incubação a 20 °C. Foi obtida uma relação linear entre o logaritmo da atividade enzimática e os diâmetros das zonas claras. O método foi validado utilizando-se soluções de alfa-amilase de cevada nas concentrações de 2,4; 60; 300 e 1.500 U.mL-1. O método tornou-se mais simples, rápido, com baixo custo e passível de ser utilizado para macrodeterminação de alfa-amilase em ampla faixa (2,4 a 7.500 U.mL-1) na investigação científica e para fins didáticos em aulas práticas.

Separación de poblaciones de glóbulos rojos senescentes por gradientes preformados de Percoll / Separation of populations of senescent red blood cells by Percoll performed gradients

Los mecanismos que determinan la senescencia de los eritrocitos han sido extensamente estudiados, sin embargo, no se han logrado conclusiones definitivas debido a la ausencia de una técnica que permita el aislamiento de grupos etáreos bien definidos. Los métodos más comúnmente empleados se basan en el aumento de densidad de los eritrocitos durante el envejecimiento. En este trabajo desarrollamos una técnica para la separación de glóbulos rojos de distintas edades empleando gradientes preformados de Percoll, un polímero sintético con propiedades fisicoquímicas adecuadas para trabajar con células vivas. En las suspensiones eritrocitarias obtenidas se realizaron determinaciones hematológicas, actividades de enzimas antioxidantes y el ensayo de eritrofagocitosis. Los valores de los parámetros hematológicos evaluados fueron significativamente mayores en las suspensiones de glóbulos rojos jóvenes. Las actividades enzimáticas mostraron una disminución de la capacidad antioxidante en las poblaciones de eritrocitos senescentes. Este proceso favorecería la interacción de los hematíes envejecidos con las células fagocíticas, demostrada median­te el ensayo de eritrofagocitosis. Los resultados obtenidos indican que el método de gradientes de Percoll permite una adecuada separación de las suspensiones eritrocitarias de distintas edades, con una eficiencia comparable a la observada en la técnica de centrifugación diferencial considerada de referencia.

The mechanisms that determine the senescence of the erythrocytes have been extensively studied; however. definitive conclusions have not been achieved mainly because of the lack of a technique that allows the isolation of well-defined etarian groups. The methods most commonly used for separating erythrocytes from different ages are based on the increase in density that these cells present during their aging. In the present work we have developed a technique for obtaining red blood cells from different ages using Percoll preformed gradients, a synthetic polymer with adequate physic-chemic properties to work with lives cells. In the erythrocytes suspensions we have made hematological determinations. activities of antioxidants enzymes and the essay of erythrophagocytosis. The values of the hematological parameters were significantly higher in the suspensions of young red blood cells. In the measurements of the enzymatic activity we observed a decrease of the antioxidant capacity in the populations of senescent erythrocytes. This process would promote the interaction between the old erythrocytes and the phagocyte cells, demonstrated by the erythrophagocytosis essay. The results obtained indicate that the method Percoll density gradients allows an appropriate separation of the erythrocytes suspensions of different ages with a comparable efficiency to that observed in the technique differential centrifugation, considered as reference.

Determinación de paraquat en orina utilizando un sistema de inyección en flujo continuo / Determination of paraquat in urine samples by flow-injection analysis

El paraguat es un herbicida que pertenece al grupo de los biperidilos. Su determinación cuantitativa en orina es muy importante para diagnosticar la supervivencia de pacientes intoxicados. Muchos centros hospitalarios utilizan pruebas semicuantitativas para la determinación de paraquat en muestras biológicas. Sin embargo, éstas suelen carecer de precisión y exactitud. Por tanto, el desarrollo de métodos alternativos simples, exactos, precisos y accesibles podría resultar muy útil en instituciones hospitalarias. Sobre la base de estas consideraciones, se propone un método de análisis por inyección en flujo y detección espectrofotométrica para la determinación cuantitativa de paraquat en muestras de orina. La determinación se basa en la formación de un producto coloreado (600 nm) posterior a la reducción de paraquat con glucosa en un medio alcalino, mediante un sistema en línea. Bajo las condiciones óptimas de operación, la ley de Beer se cumple en el intervalo 1-50 µg mL-1 de paraquat con un coeficiente de correlación >0,999. La frecuencia de análisis fue de 12 h-1 con una desviación estándar relativa del 2,8% para una solución muestra que contiene 10 µg mL-1 de paraquat (n=3). El estudio de recuperación osciló entre 97,9 y 102,1%. El método analítico fue aplicado satisfactoriamente al análisis de muestras de dos pacientes intoxicados con paraquat.

Paraguat is a herbicide beloging to the bipyridinium group.The quantitative determination of paraquat in urine of humans is very important to diagnose survival of intoxicated patients. Many hospitals use semi-quantitative tests for determining paraquat in biological samples. However, they often lack precision and accuracy. Therefore, the development of simple, precise, accurate and accessible alternative methods could be very useful in hospital institutions. Based on these considerations, a flow-injection spectrophotometric procedure is proposed for paraquat determination in urine samples. The determination is based on the formation of a coloured product (600 nm) after on-line reduction of paraquat with glucose in alkaline medium. Under optimal conditions of operation, Beer's law is obeyed in a concentration range of 1-50 µg mL-1 of paraquat with a correlation coefficient >0.999. The analytical frequency was 12 h-1 and the relative standard deviation was 2.8% for a sample solution containing 10 µg mL-1 paraquat (n=3). Recovery studies were between 97.9 and 102.1%. The analytical method was satisfactorily applied in the analyses of samples from two intoxicated patients.

Variabilidade biológica na concentração de lipídeos séricos / Biological variation in concentration of serum lipids

A variação biológica é definida como a variação natural, de ocorrência fisiológica, independente das variáveis pré-analíticas. A resposta individual aos diferentes estímulos a que o organismo é submetido resulta numa amplitude de variação biológica que oscila entre os indivíduos. Esta amplitude de variação deve ser quantificada a fim de auxiliar na interpretação dos resultados laboratoriais. O objetivo deste projeto foi verificar a variabilidade biológica na determinação de lipídeos (colesterol total, HDL colesterol, LDL colesterol e triglicerídeos) em indivíduos saudáveis. Foram convidados a participar do projeto 10 indivíduos saudáveis que foram submetidos a 10 coletas de sangue consecutivas. Após as coletas, o soro foi analisado em equipamento automatizado Cobas Mira" Marca Roche utilizando kits comerciais. As variabilidades biológicas individuais e totais encontradas foram semelhantes às descritas previamente na literatura. As menores variabilidades foram as das determinações de colesterol total (6,3%) e HDL-c (4,9%), enquanto que os triglicerídeos apresentaram o maior índice (25%). O coeficiente de variação biológica dos analitos foi superior à mediana descrita na literatura em alguns indivíduos. Nova amostra deve ser analisada quando existir discrepância entre as interpretações clínicas ou biológicas das duas amostras.

Biological variability is defined as a natural variation, with physiological occurence, independent of pre-analytical factors. The individual answer to the different stimulations to which the organism is subjected to results in degrees of biological variation that are change between subjects. This degree of variation must be quantified to help the laboratory results interpretation. The objective of this project was verify the biological variability in lipids determinations (total cholesterol, HDL cholesterol, LDL cholesterol, and triglycerides) in healthy subjects. Ten healthy individuals participated in this projetct. The samples were collected by venopunction, ten times, in three consecutive weeks. The serum obtained was analysed in automatized equipment Cobas Mira" (Roche) using commercial kits. Both individual and total variability coefficients found were similar to the literature. Total cholesterol (6.3%) and HDL-cholesterol (4.9%) showed the lowest variabilities, whereas triglycerides presented the biggest index (25%). In some patients the variation coefficient for any analyte was superior than the range previously described in the literature. New sample analyses must be performed when there is some discrepancy in clinical or biological interpretation between results.

Glicación in vitro de lipoproteínas de baja densidad y aterogenicidad en cultivos celulares / In vitro glycation of low-density lipoproteins and atherogenicity in cell culture

La hiperglucemia sostenida incrementa la glicación de proteínas. En particular, las modificaciones en las lipoproteínas de baja densidad (LDL) aumentan su potencial aterogénico. En este trabajo se comparan las modificaciones producidas por la glicación in vitro de LDL aisladas por dos métodos: precipitación selectiva (PS) y ultracentrifugación (UC). Para ello, se determinó el incremento de fructosamina, el consumo de los grupos e-amino de lisina, guanidinio de arginina y la disminución de residuos de triptofano. Para todos los analitos, los resultados cinéticos indicaron diferencias significativas con relación al basal (p<0,05), coincidentes para ambos métodos en el tiempo de aparición y en el porcentaje de variación. La aterogenicidad de las LDL glicadas separadas por PS fue estudiada en cultivos de macrófagos RAW 264.7 evaluando la formación de células espumosas y cuantificando la incorporación de LDL por tinción de los depósitos lipídicos con Oil Red. Los resultados indican que la captación de LDL modificadas aumentó con el tiempo de incubación, siendo mayor la aterogenicidad de las LDL glicadas respecto de las nativas (p<0,001, 1 h a 37 °C). El procedimiento de PS seleccionado, accesible al laboratorio bioquímico clínico, permite evaluar las modificaciones por glicación que sufren las LDL en pacientes diabéticos.

Long-term hyperglycemia increases protein glycation. In particular, modifications in the low-density lipoproteins (LDL) increase their atherogenic potential. In this study, the modifications caused by in vitro glycation of LDL separated by two methods: selective precipitation (SP) and ultracentrifugation (UC) were compared. Increase fructosamine level, decrease of e-amino group of lysine, guanidinio of arginine and triptophan fluorescence were determined. Results showed significant differences vs. basal (p<0.05) for all the tested parameters, with coincidence for the two separation methods both in time and grade of modifications. The atherogenicity of glycated LDL separated by SP was studied in macrophages RAW 264.7 in culture, through the formation of foam cells and the quantification of the dye taken up by the cellular storage lipids. Results show that the uptake of modified LDL by macrophages increased with the time of incubation, being the atherogenicity of glycated LDL greater than native LDL (p<0.001, 1 h at 37 °C). The selected SP procedure, within the facilities of routine biochemical laboratory, enables the evaluation of the modifications caused by glycation in the LDL of diabetic patients.

Comparación de las coloraciones de Giemsa y Grocott en el diagnóstico de la histoplasmosis / Comparison of Giemsa and Grocott stains in the diagnosis of histoplasmosis

El diagnóstico de histoplasmosis se realiza tradicionalmente mediante el reconocimiento de típicas levaduras intracelulares de Histoplasma capsulatum en preparaciones microscópicas teñidas con Giemsa. Se comparó la eficacia de una modificación rápida de la técnica de Grocott (MRG) y la tradicional de Giemsa para el diagnóstico de la histoplasmosis, a partir de la aplicación de ambas a 10 secreciones respiratorias, 8 escarificaciones de lesiones cutáneas y una biopsia ganglionar, pertenecientes todas a pacientes con sospecha clínica de esta micosis. En 15 de las 19 muestras no se encontraron diferencias significativas en la capacidad y rapidez para arribar al diagnóstico, mientras que en las 4 restantes, fueron reconocidas con la MRG estructuras que pasaron desapercibidas con la coloración de Giemsa. La modificación rápida permitió un reconocimiento más rápido del H. capsulatum en materiales donde este hongo se observó en escaso número y permitió además identificar con seguridad otros patógenos fúngicos diferentes de H. capsulatum, como Pneumocystis jiroveci, Paracoccidioides brasiliensis y Cryptococcus neoformans, difíciles de observar con la coloración de Giemsa. Se propone la técnica de Grocott o su modificación rápida para el diagnóstico de la histoplasmosis, especialmente cuando el empleo de la coloración de Giemsa da lugar a resultados negativos o dudosos.

The diagnosis of histoplasmosis is traditionally achieved by recognizing the typical intracellular yeasts of Histoplasma capsulatum, in smears stained with Giemsa stain. The usefulness of a rapid modification of Grocott and of traditional Giemsa stains for the diagnosis of histoplasmosis was compared applying both techniques in 10 respiratory secretions, 8 cutaneous lesions scrapings and 1 adenomegaly biopsy, all of them belonging to patients with clinically suspected histoplasmosis. In 15 out of the 19 evaluated samples, no significant differences were found in the ability or speed to reach the diagnosis with the applied techniques; while in the remaining 4 samples, structures that had not been observed with Giemsa stain were recognized with the rapid modification. The modification enabled quicker recognition of H. capsulatum than Giemsa stain in those clinical samples where the number of these fungal pathogens was scant. Additionally, the rapid modification also enabled the recognition of fungal pathogens other than H. capsulatum, as Pneumocystis jiroveci, Paracoccidioides brasiliensis and Cryptococcus neoformans, difficult to observe with the Giemsa stain. Use of Grocott technique or rapid modification stain is proposed for the diagnosis of histoplasmosis, when the result obtained with the Giemsa stain is doubtful or negative.

Studies on distribution of heavy metals in Ganga water and its bed sediments along the Patna stretch.

In the present study, the distribution of heavy metals in Ganga water and its bed sediments were studied at three sampling stations along the stretch of Patna. The samples were collected, preserved, digested with HNO3-HClO4 and analyzed by Atomic Absorption Spectrometer (AAS). The heavy metals were not detected in the dissolved form in Ganga water. The reason may be alkaline nature of water. These were detected in trace quantity in Ganga water and its bed sediments samples as total metals. The heavy metals exist in the aqueous phase due to their absorption on particulate. The sources may be attributed to geo-chemical transformation, weathering of soils and anthropogenic activities. The presence of heavy metals in the bed sediments and in the suspended phase indicates that Ganga riverine system acts as both carrier and sink for heavy metals in trace levels.

Hydrochemical investigations and correlation--analysis of ground water quality of Jaipur city, Rajasthan (India).

The objective was to investigate the quality of drinking water of Jaipur city during pre-monsoon session (April 2006 to June 2006). Physico-chemical parameters like pH, EC, TDS, Ca2+, Mg2+, Na+, K+, CO3(2-), HCO3(-), Cl(-), SO4(2-), NO3(-), F(-) and TH were analyzed by adopting the standard method of APHA. To assess the quality of ground water, each parameter was compared with the standard desirable limit of that parameter stipulated for drinking water as prescribed by BIS. A correlation analysis was conducted to determine the correlation coefficient (r) among the parameters. The highest correlation was found between EC and chloride (r = 0.986, p = < .0001). EC showed highly significant positive correlation with chloride, Mg++, Na+, TDS and TH while significant inverse correlations were found in four cases, i.e. between pH and bicarbonate, between carbonate and bicarbonate, between pH and TDS and between sulphate and pH, while potassium, nitrate and fluoride did not show any significant correlations with any other parameters studied.

TLC-spectrophometric separation and trace determination of monocrotophos and dichlorvos in enviromental and biological samples.

Organophosphorus insecticides, monocrotophos and dichlrovos are increasingly being used in agriculture to control insects on a wide range of crops. Their ready access has resulted in misuse in many instances of homicidal and suicidal poisoning cases. This paper describes about a chromogenic spray reagent for the detection/determination of monocrophos and dichlrovos in environmental and biological samples by TLC and spectrophotometric method. Monocrotophos and dichlorvos on alkaline hydrolysis yield N-methyl acetoacetamide and dichlroacetaldehyde respectively, which in turn react with diazotized p-amino acetophenone to give red-violet and red coloured compounds. Other organophosphorus insecticides do not give this reaction. Moreover, organochlorine and synthetic pyrethroid insecticides and constituents of viscera (amino acids, peptides, proteins etc), which are generally coextracted with the insecticides, do not interfere. However, phenolic compounds and hydrolysed product of carbamate insecticides may interfere and differentiate from monocrotophos and dichlrovos by Rf values. The lower limit of detection is 0.2 mg for monocrotophos and 0.1 mg for dichlorovos. The absorption maxima of the reddish-violet and red colour formed by monocrotophos and dichlrovos, are measured at 560 nm and 540 nm respectively. Beer's Law is obeyed over the concentration range of 1.2 to 6.8 mg and 6.2 to 35 mg in the final solution volume of 25 mL. The molar absorptivity and Sandell's sensitivity of monocrotophos and dichlrovos were found to be 7.1 x 10(5) (+100) 1 mole(-1) cm(-1) and 0.008 mg cm(-2), 1.2 x 10(5) 1 mole(-1) cm(-1) and 0.003 mg cm(-2) respectively. The standard deviation and relative standard deviation were found be +/- 0.005 and 2.05% +/- 0.007 and 2.02% respectively. The developed method has been successfully applied to the detection and determination of monocrotophos and dichlrovos in environmental and biological samples.

review of urinary stone analysis techniques

Knowledge of the chemical composition and structure of urinary stones is of great value in the choice of treatment and prevention of recurrence. This is a brief review and a comparative study of the principles and practical application of various chemical and physical techniques used for urinary stone analysis. The different methods of classifying and grouping urinary stones by results of chemical analytic techniques are, also, compared and evaluated. In addition to reviewing various techniques used for the in-vitro analysis of removed stone samples, the newly emerging physical and radiological techniques for the in-vivo intact-stone analysis are, also, evaluated. These in-vivo techniques, particularly the rapidly advancing unenhanced spiral CT scanning, represent an important step forward towards the notion of non- destructive analysis of urinary stones while still in situ before the choice of treatment modality