Determinación de compuestos cianogénicos amigdalina y prunasina en semillas de almendras (prunus dulcis l) utilizando cromatografía liquida de alta resolución / Determination of cyanogenic compound amygdalin and prunasin in almond kernels (prunus dulcis l) by using liquid chromatography / Determinação de compostos cianogênicos amigdalina e prunasina em sementes e amêndoa (prunus dulcis miller) cromatografia líquida de alta resolução

El objetivo del presente trabajo fue aplicar un técnica para determinar y cuantificar por separado los compuestos cianogénicos que pueden estar presentes en la semilla de almendra madura (Prunus dulcis). Entre los métodos encontrados, se seleccionó la cromatografía de líquidos de alta resolución (HPLC), que permite la cuantificación de los glucósidos cianogénicos amigdalina y prunasina por separado, adecuando diferentes procedimientos de extracción como el tamaño de partículas que influye en el proceso de liofilización, donde a menor superficie mayor área de contacto para la sublimación. Se ensayaron muestras sin grasa y con grasa, utilizando los resultados con muestras con grasa, dados los resultados obtenidos. Se utilizó metanol 100% como extractante de los glucósidos cianogénicos, resultando una concentración de amigdalina máxima a partir un tiempo de extracción de 12 horas y como fase móvil acetonitrilo/agua (20:80), se obtiene amigdalina, con una concentración de 9,8 mg/100g de muestra seca. Los cromatogramas obtenidos presentan tiempo de retención (Tr), Amigdalina: 3,4 y Prunasina, 5,7, dos picos con excelente resolución, por lo tanto las condiciones anteriores se pueden utilizar para la identificación y cuantificación de amigdalina y prunasina.

The aim of this study was to apply a technique to identify and quantify separately also cyanogenic compounds that may be present in the mature seed almond (Prunus dulcis). Among the methods selected the chromatography of liquids of high resolution (HPLC), that permit the quantification of the glycosides for the separation process of Freeze Dry where there is less surface there is more contact to sublimation without fat samples, looking at the obtain results and supported by other investigations, the use of 100 % methanol extract as a mobile phase acetonitrile-water (80:20) the results obtained of the glycosides cyanogenics resulting in a concentration of maximum amygdalin from the time of extraction of twelve hours, amygdalin is obtained, with a concentration of 9,8 mg / 100 g of dry sample. The chromatograms obtained a time of retention (Tr), amygdalin 3,4 and prunasin 5,7 two peaks with excellent resolution, to the above conditions can be used for analysis by HPLC, identification and quantification of amygdalin and prunasina.

Neste trabalho, a técnica é aplicada para determinar e também para quantificar separadamente compostos cianogénicos que podem estar presentes na semente madura amêndoa (Prunus dulcis). Métodos encontrados é seleccionado de cromatografia líquida de alta eficiência (HPLC), que permite a quantificação dos glicosídeos separar adaptar diferentes técnicas de extracção, tais como o tamanho de partícula influencia o processo de liofilização, onde a área de superfície maior menor sublimação contacto com desengradas amostras de gordura e usando os resultados com amostras desengorduradas, Tendo em vista os resultados obtidos, e suportados por outras pesquisas metanol a 100 % foi usado como o agente de extracção e como fase móvel acetonitrilo/água (80:20) de glicósidos cianogénicos, resultando numa concentração elevada de amigdalina a partir de um tempo de extracção de 12 horas. Amigdalina é obtido, com uma concentração de 9,8 mg / 100 g de amostra seca. Os cromatogramas apresentados tempo de retenção (Tr), Amygdalin: 3,4 e prunasina 5,7 dois picos com excelente resolução, com as condições acima podem ser utilizados para a análise por HPLC. identificação e quantificação de amigdalina e prunasina.

Determinación d compuestos cianogénicos amigdaina y prunasina en semillas de almendras (prunus dulcis l) utilizando cromatografía liquida de alta resolución / Determination of cyanogenic compound amygdalin and prunasin in alond kernels (prrunus dulcis l) by using liquid chromatography / Detrminação de ccompostos cianogênicos amigdalina eprunasina em seementes e amêndoa (prunus dulcis miller) cromatografia líquida de alta resolução

El objetivo del presente trabajo fue aplicar un técnica para determinar y cuantificar por separado los compuestos cianogénicos que pueden estar presentes en la semilla de almendra madura (Prunus dulcis). Entre los métodos encontrados, se seleccionó la cromatografía de líquidos de alta resolución (HPLC), que permite la cuantificación de los glucósidos cianogénicos amigdalina y prunasina por separado, adecuando diferentes procedimientos de extracción como el tamaño de partículas que influye en el proceso de liofilización, donde a menor superficie mayor área de contacto para la sublimación. Se ensayaron muestras sin grasa y con grasa, utilizando los resultados con muestras con grasa, dados los resultados obtenidos. Se utilizó metanol 100% como extractante de los glucósidos cianogénicos, resultando una concentración de amigdalina máxima a partir un tiempo de extracción de 12 horas y como fase móvil acetonitrilo/agua (20:80), se obtiene amigdalina, con una concentración de 9,8 mg/100g de muestra seca. Los cromatogramas obtenidos presentan tiempo de retención (Tr), Amigdalina: 3,4 y Prunasina, 5,7, dos picos con excelente resolución, por lo tanto las condiciones anteriores se pueden utilizar para la identificación y cuantificación de amigdalina y prunasina.

The aim of this study was to apply a technique to identify and quantify separately also cyanogenic compounds that may be present in the mature seed almond (Prunus dulcis). Among the methods selected the chromatography of liquids of high resolution (HPLC), that permit the quantification of the glycosides for the separation process of Freeze Dry where there is less surface there is more contact to sublimation without fat samples, looking at the obtain results and supported by other investigations, the use of 100 % methanol extract as a mobile phase acetonitrile-water (80:20) the results obtained of the glycosides cyanogenics resulting in a concentration of maximum amygdalin from the time of extraction of twelve hours, amygdalin is obtained, with a concentration of 9,8 mg / 100 g of dry sample. The chromatograms obtained a time of retention (Tr), amygdalin 3,4 and prunasin 5,7 two peaks with excellent resolution, to the above conditions can be used for analysis by HPLC, identification and quantification of amygdalin and prunasina.

Neste trabalho, a técnica é aplicada para determinar e também para quantificar separadamente compostos cianogénicos que podem estar presentes na semente madura amêndoa (Prunus dulcis). Métodos encontrados é seleccionado de cromatografia líquida de alta eficiência (HPLC), que permite a quantificação dos glicosídeos separar adaptar diferentes técnicas de extracção, tais como o tamanho de partícula influencia o processo de liofilização, onde a área de superfície maior menor sublimação contacto com desengradas amostras de gordura e usando os resultados com amostras desengorduradas, Tendo em vista os resultados obtidos, e suportados por outras pesquisas metanol a 100 % foi usado como o agente de extracção e como fase móvel acetonitrilo/água (80:20) de glicósidos cianogénicos, resultando numa concentração elevada de amigdalina a partir de um tempo de extracção de 12 horas. Amigdalina é obtido, com uma concentração de 9,8 mg / 100 g de amostra seca. Os cromatogramas apresentados tempo de retenção (Tr), Amygdalin: 3,4 e prunasina 5,7 dois picos com excelente resolução, com as condições acima podem ser utilizados para a análise por HPLC. identificação e quantificação de amigdalina e prunasina.

Prevención de la hemorragia posparto en una gestante afectada de síndrome de Hermansky-Pudlack / Prevention of postpartum hemorrhage in a patient with Hermansky-Pudlak syndrome

El síndrome de Hermansky-Pudlak es una enfermedad multisistémica de herencia autosómica recesiva. Se caracteriza principalmente por albinismo óculo-cutáneo y alteración de la agregación plaquetaria. Se presentan el seguimiento y la finalización de la gestación de una paciente de 30 años afectada de dicho síndrome. Se describen las medidas profilácticas realizadas durante el trabajo de parto con el objetivo de evitar complicaciones hemorrágicas debidas a la disfunción plaquetaria. La finalización de la gestación tuvo lugar a las 38,2 semanas mediante un parto eutócico sin anestesia peridural y con buena evolución materno-fetal(AU)

Hermansky-Pudlak syndrome is a multisystemic disease with autosomal recessive inheritance, mainly characterized by oculo-cutaneous albinism and impaired platelet aggregation. We describe the follow–up and end of pregnancy in a 30-year-old woman with this syndrome, as well as the measures carried out during labor to avoid bleeding complications due to platelet dysfunction. The pregnancy ended at 38.2 weeks through vaginal delivery, without epidural anesthesia and good maternal and fetal outcome(AU)

Heparinized solution vs. saline solution in the maintenance of arterial catheters: a double blind randomized clinical trial.

OBJECTIVES: The objectives were to analyze the effectiveness of heparinized solution vs. saline solution for the maintenance of arterial catheters and to detect changes in the activated partial thromboplastin time (aPTT) and platelet count in the samples extracted from both groups of arterial catheters. DESIGN: Randomized, double blind, placebo-controlled clinical trial. SETTING: Intensive Care Unit of a third-level hospital in Terrassa, Barcelona, Spain. PATIENTS: One hundred and thirty-three patients were included in the trial. The selection criteria were: adults, informed consent, not receiving either full-dose anticoagulant or fibrinolytic treatment, and no thrombocytopenia. INTERVENTIONS: Sixty-five patients received heparinized solution (1 IU/ml) and 68 received saline solution. MEASUREMENTS: Arterial catheter functionality was compared in the groups every 8 h and at catheter removal. Patency, reliability of arterial pressure, and curve quality were used to evaluate the functionality of the catheters. Blood was drawn, discarding 7.5 ml, from the arterial catheter and from the venouscatheter simultaneously for coagulation tests. RESULTS: The median duration of catheters being in place was 5.1 days (IQR = 8.1) in the heparin group, and 5.4 (IQR = 7.3) in the saline group (p = 0.7). Kaplan-Meier curves showed no differences between groups (p = 0.6). The number of manipulations required to maintain the patency of the arterial catheters was 35% vs. 40% (p = 0.5). The heparin group had a significantly longer aPTT (2.1 +/- .3 vs. 1.25 +/- 0.3, p = 0.001). CONCLUSIONS: The use of heparinized solution for arterial catheter maintenance doesnot appear to be justified. It did not increase the duration of the catheters, nor did it improve their functionality significantly. On the other hand, heparin Na altered aPTT significantly.

Highly active antiretroviral therapy and incidence of cervical squamous intraepithelial lesions among HIV-infected women with normal cytology and CD4 counts above 350 cells/mm3.

OBJECTIVES: To provide evidence for the long-term effect of highly active antiretroviral therapy (HAART) on the incidence of cervical squamous intraepithelial lesions (SILs) among HIV-positive women with normal cytology test and CD4 count above 350 cells/mm(3). PATIENTS AND METHODS: A retrospective cohort study was carried out in HIV-positive women with two consecutive normal cervical cytological tests (Papanicolaou test) and at least one subsequent test, without previous cervical history of SIL or cancer diagnosis, and with an immunological status >350 CD4 cells/mm(3). The patients were divided into two groups: treated with HAART (HAART group) or not treated with HAART (NO-HAART group), during the period of time between cytology tests included in the survival analysis and time until SIL. RESULTS: Between January 1997 and December 2006, 127 women were included: 90 in the HAART group and 37 in the NO-HAART group. Both groups of patients were similar with respect to demographic data, except for HIV viral load and previous HAART inclusion (P < 0.001). SIL was diagnosed in 27 of 90 (30%) patients in the HAART group and in 7 of 37 (19%) patients in the NO-HAART group (OR = 1.84, 95% CI: 0.72-4.69, P = 0.202). The actuarial probability of remaining free of SIL at 3 years was 70% in the HAART group and 78% in the NO-HAART group. No variable was associated with an increased risk of developing SILs. CONCLUSIONS: These results suggest that when the patients' immunological status is above 350 CD4 cells/mm(3), the HIV-infected women treated with HAART present a similar cervical SIL incidence to women not on HAART.

Evolution of cervical cytologic changes among HIV-infected women with normal cytology in the HAART era.

The influence of HAART on the evolution to cervical squamous intraepithelial lesions (SIL) among HIV(+) women with a normal cytological test in the HAART era was studied. A retrospective cohort study (1997-2005) of HIV-infected women treated with HAART was conducted. Those with a normal cervical cytology (Papanicolaou test) and at least one subsequent test were included. Survival (time until diagnosis of SIL), univariate, and multivariate analyses were performed. A total of 133 HIV-infected patients treated with HAART were included. The incidence of SIL was 35% (47 patients). SIL was diagnosed in 36 of 110 (33%) patients with a baseline and final immunological status of >200 CD4 cells/microl and in 6 of 9 (67%) patients with a baseline and final immunological status of < or =200 CD4 (OR: 0.24, 95% CI: 0.06-1.03, p = 0.041). SIL was diagnosed in 10 of 60 (17%) patients with an undetectable baseline and final HIV viral load and in 36 of 70 (51%) patients with a detectable HIV viral load (OR: 0.19, 95% CI: 0.07-0.46, p < 0.001). A high incidence of SIL (cancer precursor lesions) was observed among HIV(+) women without a background of cervical pathology. The effect of HAART on the control of HIV replication and of immunological status (>200 CD4) through the follow-up was associated with a reduction of SIL.

Microwave-assisted headspace single-drop microextration of chlorobenzenes from water samples.

A one-step and in-situ sample preparation method used for quantifying chlorobenzene compounds in water samples has been developed, coupling microwave and headspace single-drop microextraction (MW-HS-SDME). The chlorobenzenes in water samples were extracted directly onto an ionic liquid single-drop in headspace mode under the aid of microwave radiation. For optimization, a Plackett-Burman screening design was initially used, followed by a mixed-level factorial design. The factors considered were: drop volume, aqueous sample volume, stirring speed, ionic strength, extraction time, ionic liquid type, microwave power and length of the Y-shaped glass-tube. The optimum experimental conditions found from this statistical evaluation were: a 5 microL microdrop of 1-hexyl-3-methylimidazolium hexafluorophosphate exposed for 20 min to the headspace of a 30 mL aqueous sample, irradiated by microwaves at 200 W and placed in a 50 mL spherical flask connected to a 25 cm Y-shaped glass-tube. Under the optimised experimental conditions, the response of a high performance liquid chromatographic system was found to be linear over the range studied and with correlation coefficients ranging between 0.9995 and 0.9999. The method showed a good level of repeatability, with relative standard deviations varying between 2.3 and 8.3% (n=5). Detection limits were found in the low microg L(-1) range varying between 0.016 and 0.039 microg L(-1). Overall, the performance of the proposed method demonstrated the favourable effect of microwave sample irradiation upon HS-SDME. Finally, recovery studies from different types of environmental water samples revealed that matrix had little effect upon extraction.

An ionic liquid as a solvent for headspace single drop microextraction of chlorobenzenes from water samples.

A headspace single-drop microextraction (HS-SDME) procedure using room temperature ionic liquid and coupled to high-performance liquid chromatography capable of quantifying trace amounts of chlorobenzenes in environmental water samples is proposed. A Plackett-Burman design for screening was carried out in order to determine the significant experimental conditions affecting the HS-SDME process (namely drop volume, aqueous sample volume, stirring speed, ionic strength, extraction time and temperature), and then a central composite design was used to optimize the significant conditions. The optimum experimental conditions found from this statistical evaluation were: a 5 microL microdrop of 1-butyl-3-methylimidazolium hexafluorophosphate, exposed for 37 min to the headspace of a 10 mL aqueous sample placed in a 15 mL vial, stirred at 1580 rpm at room temperature and containing 30% (w/v) NaCl. The calculated calibration curves gave a high level of linearity for all target analytes with correlation coefficients ranging between 0.9981 and 0.9997. The repeatability of the proposed method, expressed as relative standard deviation, varied between 1.6 and 5.1% (n=5). The limits of detection ranged between 0.102 and 0.203 microg L(-1). Matrix effects upon extraction were evaluated by analysing spiked tap and river water as well as effluent water samples originating from a municipal wastewater treatment plant.

[Contribution of human papillomavirus second-generation hybrid capture test for the diagnosis of cervical pathology in HIV-infected outpatients]. / Contribución de la captura de híbridos de segunda generación del virus del papiloma humano en el cribado de afección cervical en mujeres con infección por el virus de la inmunodeficiencia humana.

BACKGROUND AND OBJECTIVE: The causal relationship between human papillomavirus (HPV) and cervical cancer is well established. The initial diagnosis of HPV-related cervical infection is currently performed by HPV-associated changes in cervical cytology. We aimed to study the accuracy and concordance between HPV ADN detection by second-generation hybrid capture (HC-2) and cervical cytological changes for the diagnosis of HPV cervical infection in human immunodeficiency virus (HIV+) outpatients. PATIENTS AND METHOD: From March 1999 to August 2002, 139 HIV+ patients were included. HPV infection was determined by cytology and HC-2. The accuracy and level of concordance between both techniques was analyzed. RESULTS: The applicability of the HC-2 test was 96%. Sixty-eight (49%) patients were diagnosed with HPV infection by HC-2. High-oncogenetic-risk HPV genotypes were detected in 64 (46%) patients. The sensitivity, specificity and positive and negative predictive values of HC-2 in HPV detection were 78%, 69%, 61% and 83%, respectively. The concordance was K = 0.44 (95% confidence interval, 0.29-0.60); p < 0.001. CONCLUSIONS: The HC-2 diagnostic technique for HPV-related cervical infection in HIV+ patients is a sensitive and specific test. The combined use of both tests might increase the diagnostic efficacy, and hence have positive repercussions on cervical pathology screening on an outpatient basis.

Contribución de la captura de híbridos de segunda generación del virus del papiloma humano en el cribado de afección cervical en mujeres con infección por el virus de la inmunodeficiencia humana / Contribution of human papillomavirus second-generation hybrid capture test for the diagnosis of cervical pathology in HIV-infected outpatients

Fundamento y objetivo: El papel etiológico del virus del papiloma humano (VPH) en las neoplasias invasivas de cérvix uterino está demostrado. Aunque primero se produce la infección por el VPH y después los cambios histopatológicos consecuencia de la infección, en la práctica clínica el cribado de la enfermedad neoplásica cervical se basa inicialmente en los cambios citopatológicos. El objetivo de este estudio ha sido estudiar la precisión de la técnica de detección del ADN del VPH por captura de híbridos de segunda generación (CH-2) respecto a la citología cervical y la concordancia entre ambas técnicas en el cribado de la infección por el VPH en mujeres infectadas por el virus de la inmunodeficiencia humana (VIH) controladas ambulatoriamente. Pacientes y método: Durante el período comprendido entre marzo de 1999 y agosto de 2002 se incluyó de forma prospectiva a 139 pacientes infectadas por el VIH que acudieron a la visita ambulatoria. El cribado de la infección por el VPH se exploró mediante citología cervical y CH-2. Se estudió la precisión de la CH-2 respecto a la citología cervical y el grado de concordancia entre ambas técnicas. Resultados: Se aplicó la CH-2 en 135 pacientes (96%), de las cuales 68 (49%) tuvieron un resultado positivo para el VPH. Se detectó infección por genotipos de alto riesgo oncogénico en 64 pacientes (46%). La sensibilidad, especificidad, el valor predictivo positivo y negativo de la CH-2 en la detección del VPH fueron del 78, el 69, el 61 y el 83%, respectivamente. Ambas técnicas presentaron una concordancia de kappa de 0,44 (intervalo de confianza del 95%: 0,29-0,60; p < 0,001). Conclusiones: La CH-2 es un método sensible y específico en el cribado de la infección por el VPH en pacientes infectadas por el VIH. La combinación de citología cervical y captura de híbridos en el cribado de la neoplasia cervical puede ayudar a reducir la incidencia de cáncer de cuello uterino en las pacientes con infección por el VIH

Background and objective: The causal relationship between human papillomavirus (HPV) and cervical cancer is well established. The initial diagnosis of HPV-related cervical infection is currently performed by HPV-associated changes in cervical cytology. We aimed to study the accuracy and concordance between HPV ADN detection by second-generation hybrid capture (HC-2) and cervical cytological changes for the diagnosis of HPV cervical infection in human immunodeficiency virus (HIV+) outpatients. Patients and method: From March 1999 to August 2002, 139 HIV+ patients were included. HPV infection was determined by cytology and HC-2. The accuracy and level of concordance between both techniques was analyzed. Results: The applicability of the HC-2 test was 96%. Sixty-eight (49%) patients were diagnosed with HPV infection by HC-2. High-oncogenetic- risk HPV genotypes were detected in 64 (46%) patients. The sensitivity, specificity and positive and negative predictive values of HC-2 in HPV detection were 78%, 69%, 61% and 83%, respectively. The concordance was K = 0.44 (95% confidence interval, 0.29-0.60); p < 0.001. Conclusions: The HC-2 diagnostic technique for HPV-related cervical infection in HIV+ patients is a sensitive and specific test. The combined use of both tests might increase the diagnostic efficacy, and hence have positive repercussions on cervical pathology screening on an outpatient basis