Effect of temperature and cell concentration on dolphin (Tursiops truncatus) spermatozoa quality evaluated at different days of refrigeration.

The use of cryopreserved dolphin spermatozoa facilitates the exchange of genetic material between aquatic parks and makes spermatozoa accessible to laboratories for studies to further our understanding of marine mammal reproduction. Sperm cryopreservation in the bottlenose dolphin (Tursiops truncatus) has been developed for the exchange of gametes within the ex situ population. The aim of this study was to develop an effective method for refrigeration of bottlenose dolphin spermatozoa diluted in a commercial extender (BTS). In Experiment 1, the effect of temperature (5 compared with 15 °C) on sperm quality was evaluated during 7 days of storage at 100 × 106 spermatozoa/ml. In Experiment 2, the effect of the storage concentration (100 × 106 compared with 20 × 106 spermatozoa/ml) on sperm quality was assessed during 7 days of storage at 5 °C. In Experiment 1, total motility (including % of rapid sperm) was greater at 5 than 15 °C. When the effect of storage concentration was evaluated (Experiment 2), total motility and ALH were greater at the higher storage concentration (100 × 106 spermatozoa/ml). For both experiments, values for viability, acrosome integrity, and normal morphology variables were consistent throughout the 7 days of refrigeration. In Experiment 3, a microbiological study was performed to evaluate the effect of the refrigeration temperature and days of storage on bacterial growth. The results of microbiological analysis indicated there was Staphylococcus aureus in some samples, however, there was no effect of temperature or days of refrigeration. In conclusion, bottlenose dolphin semen can be refrigerated for a short to medium period of storage and there is maintenance of functionality of sperm when stored at 100 × 106 spermatozoa/ml at 5 °C.

Prevalencia, grado de control y tratamiento de la hipertensión arterial en al población de 30 a 74 años de la Comunidad de Madrid. estudio Predimerc / Prevalence, degree of control and treatment of hypertension in the adult population of Madrid, Spain

Fundamentos: La hipertensión arterial (HTA) es uno de los principales factores de riesgo cardiovascular modificables. El objetivo del estudio es estimar en la población de 30 a 74 años de la Comunidad de Madrid (CM) la prevalencia de hipertensión arterial, el nivel de conocimiento y tratamiento farmacológico, y grado de control. Métodos: Estudio transversal de una muestra poblacional en 2007 entre los residentes en la Comunidad Madrid de entre 30 y 74 años, obtenida por muestreo aleatorio por conglomerados bietápico. Se aplicó un cuestionario epidemiológico por vía telefónica. En el centro de salud se realizó la exploración física por un equipo de enfermeras entrenadas. Para las variables cuantitativas se han calculado las medias e intervalos de confianza del 95% (IC del 95%) y para las cualitativas el porcentaje y el IC del 95% . Para evaluar el control de la HTA según número de principios activos se calcularon las Odds ratio mediante regresión logística. Resultados: La muestra definitiva incluyó a 1.085 hombres y 1.183 mujeres, con una edad media de 48,3 años. La prevalencia de hipertensión fue de 29,3% (IC95%:27,3-31,5) (35,1% en hombres y 23,9% en mujeres (p<0,01). De las 665 personas hipertensas detectadas el 68,6% conocía su condición (62,4% los varones y 76,65 las mujeres, p<0,05) de los cuales el 54% estaba en tratamiento farmacológico (45,4% los varones y 64,5% las mujeres, p<0,05). De las 265 personas hipertensas en tratamiento tenían controlada la tensión arterial el 33% de los varones frente al 49% de las mujeres (p<0,05). Las personas tratadas con dos o más principios activos mejoraron su control frente a las tratadas con monoterapia [ORa: 1,89; (IC95%:1,09-33,27)]. Conclusiones: Nuestras cifras de prevalencia, tratamiento y control ocupan un lugar intermedio-bajo entre las publicadas en estudios previos sobre poblaciones españolas y europeas. Sin embargo la HTA es un problema de salud pública en la CM. Un porcentaje importante de personas hipertensas no están diagnosticadas. De los hipertensos que conocen su diagnóstico y reciben tratamiento sólo 1 de cada 3 varones y una de cada 2 mujeres están bien controlados(AU)

Background: Hypertension (HT) is a major modifiable cardiovascular risk factors. The objective of this study is to estimate the adult population of the Community of Madrid (30-74 years) prevalence of hypertension, knowledge, degree of control and drug treatment. Methods: We studied a random sample population in 2007 among residents of Madrid Community of between 30 and 74 years, obtained by cluster sampling. An epidemiological questionnaire was applied through telephone interview. Physical examination was conducted by a team of trained nurses in the health center. . ; To assess control of hypertension by number of active ingredients Odds were calculated through logistic regression models. Results: The final sample included 1,085 men and 1,183 women with a mean age of 48.3 years. The prevalence of hypertension was 29.3% (35.1% men and 23.9% in women (p <0.01). Of the 665 hypertensives identified 68.6% knew their condition (62.4 76.65% in males and females, p <0.05), of which 54% were in drug treatment (45.4% in men and 64.5% in women, p <0.05). Of the 265 treated hypertensive patients, blood pressure was controlled in 33% of males and 49% of women, p <0.05. Patients treated with two or more active improves your control (aOR: 1, 89). Conclusions: Our data on prevalence, treatment and control are at the lower-intermediate between those reported in previous studies on Spanish and European populations. However, the HA is a public health problem in the CM. Many hypertensive patients are not detected. Of those detected, only 1 in3 of men and 1 in 2 women are well controlled(AU)

SKP2 oncogene is a direct MYC target gene and MYC down-regulates p27(KIP1) through SKP2 in human leukemia cells.

SKP2 is the ubiquitin ligase subunit that targets p27(KIP1) (p27) for degradation. SKP2 is induced in the G(1)-S transit of the cell cycle, is frequently overexpressed in human cancer, and displays transformation activity in experimental models. Here we show that MYC induces SKP2 expression at the mRNA and protein levels in human myeloid leukemia K562 cells with conditional MYC expression. Importantly, in these systems, induction of MYC did not activate cell proliferation, ruling out SKP2 up-regulation as a consequence of cell cycle entry. MYC-dependent SKP2 expression was also detected in other cell types such as lymphoid, fibroblastic, and epithelial cell lines. MYC induced SKP2 mRNA expression in the absence of protein synthesis and activated the SKP2 promoter in luciferase reporter assays. With chromatin immunoprecipitation assays, MYC was detected bound to a region of human SKP2 gene promoter that includes E-boxes. The K562 cell line derives from human chronic myeloid leukemia. In a cohort of chronic myeloid leukemia bone marrow samples, we found a correlation between MYC and SKP2 mRNA levels. Analysis of cancer expression databases also indicated a correlation between MYC and SKP2 expression in lymphoma. Finally, MYC-induced SKP2 expression resulted in a decrease in p27 protein in K562 cells. Moreover, silencing of SKP2 abrogated the MYC-mediated down-regulation of p27. Our data show that SKP2 is a direct MYC target gene and that MYC-mediated SKP2 induction leads to reduced p27 levels. The results suggest the induction of SKP2 oncogene as a new mechanism for MYC-dependent transformation.

Voltammetric behaviour and square-wave voltammetric determination of the potent antioxidant and anticarcinogenic agent ellagic acid in foodstuffs.

The voltammetric behaviour of ellagic acid (EA) is investigated by cyclic, differential pulse and square-wave voltammetry (CV, DPV and SWV, respectively). Based on the anodic oxidation peak at approximately 0.42V in acetic/acetate buffer (pH 5.5) a robust and a highly reliable square-wave voltammetric method is presented for the determination of EA. The oxidation peak current was linearly dependent on the concentration of EA in the range of 1.0×10(-7)-1.5×10(-6)mol/L (r=0.9997), with a detection limit of 1.0×10(-8)mol/L (S/N=3) and a quantification limit of 3.4×10(-8)mol/L (S/N=10), good reproducibility and a satisfactory level of selectivity towards others polyphenols. The proposed method was applied to the determination of free and total EA in fruits, nuts and juices with good analytical results being obtained.

Bortezomib decreases Rb phosphorylation and induces caspase-dependent apoptosis in Imatinib-sensitive and -resistant Bcr-Abl1-expressing cells.

The use of c-abl-specific inhibitors such as Imatinib (IM) or Dasatinib has revolutionized the treatment of chronic myeloid leukemia (CML). However, a significant percentage of patients become resistant to IM. In this report, we have analyzed the possibility of using the proteasome as a molecular target in CML. Our results show that cells that express Bcr-Abl1 are more sensitive to the inhibition of the proteasome with Bortezomib (Btz) than control cells. This treatment reduces the proliferation of Bcr-Abl1-expressing cells, by inactivating NF-kappaB2 and decreasing the phosphorylation of Rb, eventually leading to an increase in caspase-dependent apoptosis. Furthermore, we show that Btz also induces cell-cycle arrest and apoptosis in cells expressing Bcr-Abl1 mutants that are resistant to IM. These results unravel a new molecular target of Btz, that is the Rb pathway, and open new possibilities in the treatment of CML especially for patients that become resistant to IM because of the presence of the T315I mutation.

Dynamic infrared imaging of cutaneous melanoma and normal skin in patients treated with BNCT.

We recently initiated a program aimed to investigate the suitability of dynamic infrared imaging for following-up nodular melanoma patients treated with BNCT. The reason that makes infrared imaging attractive is the fact that it constitutes a functional and non-invasive imaging method, providing information on the normal and abnormal physiologic response of the nervous and vascular systems, as well as the local metabolic rate and inflammatory processes that ultimately appear as differences in the skin temperature. An infrared camera, with a focal plane array of 320 x 240 uncooled ferroelectric detectors is employed, which provides a video stream of the infrared emission in the 7-14 microm wavelength band. A double blackbody is used as reference for absolute temperature calibration. After following a protocol for patient preparation and acclimatization, a basal study is performed. Subsequently, the anatomic region of interest is subjected to a provocation test (a cold stimulus), which induces an autonomic vasoconstriction reflex in normal structures, thus enhancing the thermal contrast due to the differences in the vasculature of the different skin regions. Radiation erythema reactions and melanoma nodules possess typically a faster temperature recovery than healthy, non-irradiated skin. However, some other non-pathological structures are also detectable by infrared imaging, (e.g. scars, vessels, arteriovenous anastomoses and injuries), thus requiring a multi-study comparison in order to discriminate the tumor signal. Besides the superficial nodules, which are readily noticeable by infrared imaging, we have detected thermal signals that are coincident with the location of non-palpable nodules, which are observable by CT and ultrasound. Diffuse regions of fast temperature recovery after a cold stimulus were observed between the third and sixth weeks post-BNCT, concurrent with the clinical manifestation of radiation erythema. The location of the erythematous visible and infrared regions is consistent with the 3D dosimetry calculations.

[Accumulated experience in the analysis of quality indicators in a haemodialysis unit]. / Experiencia acumulada en el análisis de indicadores de calidad en una unidad de hemodiálisis.

INTRODUCTION: The object of this work was to review our use of various indicators of haemodialysis within the guidelines of good clinical practice. MATERIALS AND METHODS: The study includes all patients from our haemodialysis program from June 2005 to February 2008. The indicators we evaluated included various areas: anaemia, bone-mineral metabolism, dialysis dose, cardiovascular risk, vascular access and morbidity/ mortality. The measurement intervals varied according to the parameter being evaluated. RESULTS: We gathered those indicators in which we found a difference between our results and the targets set. It is possible to reach a haemoglobin of > or = 11 g/dl in more than 85% of the patients, although more than 20% of them showed > 13 g/dl. We were able to stay on target with phosphorous (> 85%) but not with calcium (72.7%) or PTH (38.8%) although average values were improved. The incorporation of new patients to the haemodialysis programme, some previously unknown, limited our possibilities of achieving > or = 85% with a Kt/V > or = 1.3. Only 62.2% of the patients had a systolic blood pressure of < or = 140 mmHg. The percentage of patients dialysed by catheter (objective < 10%) was only achieved in five out of the eleven measures. The hospitalisation rate was between 0.49 and 0.71 episodes/patient/year. The patient survival rates coincide with those of the Comunidad Valenciana Register. CONCLUSION: the use of clinical performance measures has improved our results, whilst in some cases it has raised doubts over their definition and established targets. In general we feel that they should be revised and redefined where necessary in an attempt to avoid variability, iatrogenia, and increased costs. The use of only those indicators in which a clear scientific basis is evident, should be considered.

Application of a trazodone-selective electrode to pharmaceutical quality control and urine analyses.

A trazodone-selective electrode for application in pharmaceutical quality control and urine analysis was developed. The electrode is based on incorporation of a trazodone-tetraphenylborate ion exchanger in a poly(vinyl chloride) membrane. The electrode showed a fast, stable and Nernstian response over a wide trazodone concentration range (5 x 10(-5)-1 x 10(-2) M) with a mean slope of 59.3 +/- 0.9 mV/dec of concentration, a mean detection limit of 1.8 x 10(-5) +/- 2.2 x 10(-6) M, a wide working pH range (5-7.5) and a fast response time (less than 20 s). The electrode also showed good accuracy, repeatability, reproducibility and selectivity with respect to some inorganic and organic compounds, including the main trazodone metabolite. The electrode provided good analytical results in the determination of trazodone in pharmaceuticals and spiked urine samples; no extraction steps were necessary. Dissolution testing of trazodone tablets, in different conditions of pH and particle size, based on a direct potentiometric determination with the new selective electrode is presented as well.

Incidencia y prevalencia de cardiopatía isquémica y enfermedad cerebrovascular en España: revisión sistemática de la literatura / Incidence and prevalence of ischaemic heart disease and cerebrovascular disease in Spain: a systematic review of the literature

La morbilidad de las enfermedades cardiovasculares no es bienconocida en España. Esta carencia de información se debe a que noexiste una fuente de datos que sea estable, exhaustiva y fiable. Elobjetivo de este trabajo es describir la frecuencia poblacional de cardiopatíaisquémica y enfermedad cerebrovascular en España. Seidentificaron como fuentes de información los estudios científicospublicados en revistas médicas españolas o por autores españolescon fecha posterior al año 1990, mediante estrategias de búsquedabibliográfica en cuatro bases de datos distintas. Se seleccionaron losestudios derivados de registros poblacionales, encuestas de prevalenciay estudios de cohortes de los que fue posible extraer tasas deincidencia y/o prevalencia ajustadas por edad. Se identificaron 19trabajos publicados entre 1993 y 2005, con información válida sobreincidencia o prevalencia de cardiopatía isquémica o enfermedadcerebrovascular en España a nivel poblacional. Morbilidad por cardiopatíaisquémica: Las tasas de incidencia de infarto agudo de miocardiooscilaron entre 135-210 nuevos casos anuales por cada100.000 varones y entre 29-61 por cada 100.000 mujeres entre 25 y74 años. No existen datos de prevalencia de cardiopatía isquémica,solamente hay un estudio de prevalencia de angina en la que ésta seestima en el 7,3% en hombres y 7,5% en mujeres. Las tasas de incidenciade enfermedad cerebrovascular por 100.000 habitantes seestiman en 364 en hombres y 169 en mujeres. Si la incidencia semidiera en población mayor de 69 años las tasas se elevarían a 2.371en hombres y 1.493 en mujeres. La prevalencia de ictus en poblaciónmayor de 65 años en España podría estimarse en un 7% en hombresy un 6% en mujeres. La información disponible en la actualidad puedeservir como referencia inicial, pero sería necesario un esfuerzo deestandarización metodológica y de coordinación que excede la capacidadde grupos de investigación aislados

Cardiovascular disease morbidity is not well documented inSpain, due to the fact that there is no comprehensive and reliabledata source. This study sought to describe the population frequencyof ischaemic heart disease and cerebrovascular disease in Spain. Weused bibliographic search strategies in four different databases toidentify scientific studies published in Spanish medical journals orby Spanish authors after 1990. We then selected studies drawn frompopulation-based registers, prevalence surveys and cohort studies,from which age-adjusted incidence and/or prevalence rates could beextracted. We identified 19 papers published in the period 1993-2005, containing valid information on cardiovascular diseases populationincidence or prevalence in Spain. Acute myocardial infarctionincidence rates per 100,000 population ranged between 135-210 and29-61 new cases annually for men and women aged 25-74 years, respectively.Acute myocardial infarction prevalence rates were notavailable, but a study of prevalence of angina showed rates of 7,3%in men and 7,5% in women. Cerebrovascular disease incidence ratesper 100,000 population ranged between 364 for men and 169women, in aged 69 years and over rising to 2.371 in men and 1.493in women. Prevalence rates in population aged 65 years and overwould be estimated in 7% in men and 6% in women. Conclusions:Although currently available information may serve as an initialreference, monitoring of the incidence and prevalence of these diseaseswill nevertheless call for an effort which exceeds the capacity ofisolated research teams

[Vascular access outcome in patients aged over 75 years]. / Resultados del acceso vascular en mayores de 75 anos.

UNLABELLED: We report a retrospective study on the results of 132 native fistulas, 12 grafts and 27 tunneled catheters followed during 30 months in 144 patients on hemodialysis. The results were compared according to patient age: 75 years or over (n = 58, 80.3 +/- 3.5) vs below 75 years (n = 86, 59.5 +/- 13.3). Gender, presence of diabetes and type of fistula were also included in the analysis. RESULTS: There were no statistically significant differences between both groups in the use of tunneled catheters or grafts (8.6% vs 5.8% y 5.2% vs 10.5% respectively), primary failure of native fistulas (7.1% in those aged 75 years or over vs 25.5% in patients below 75 years), rate of thrombosis (0.03 vs 0.09/patient year at risk respectively) or number of percutaneous or surgical procedures in order to maintain the fistula patency (0.11 vs 0.16/patient year at risk respectively). At the same time no differences were seen in the primary, primary assisted and secondary patency of the native fistulas. The mean age of the patients when the first access fistula was created was different according to the area of surgery (74.9 +/- 9.3 for the elbow vs 64.9 +/- 16.2 years for the forearm, p < 0.005). Diabetes was an unfavourable factor for primary (HR Cox 2.08, p < 0.05) or secondary (Log Rank, p <0.05) patency. CONCLUSION: The vascular access for hemodialysis in elderly patients presents a similar evolution to that seen in younger populations if the access creation is based on an exhaustive study, including ecodoppler of the vascular map and the use of more proximal fistulas if necessary. Therefore the more frequent use of grafts or catheters in elderly patients is not justified.

[Prevalence of hyperhomocysteinemia and associated factors in primary health care]. / Prevalencia de la hiperhomocisteinemia y factores asociados en atención primaria.

BACKGROUND AND OBJECTIVE: To identify plasma homocysteine concentrations that could be taken as normal values in our population, and to measure hyperhomocysteinemia prevalence. SUBJECTS AND METHOD: Cross-sectional study performed in all subjects attending or working in a primary health care center for any reason. Information was collected about personal history, cardiovascular risk factors and socio-demografic variables, and plasma homocysteine levels and other biochemical parameters were measured. Distribution of homocysteine concentration was compared in individuals with a history of cardiovascular disease and in a gold-standard population (healthy subjects with normal serum concentrations of B-vitamins). The value of homocysteine concentration chosen as cut-off value was the one that offered an optimal sensitivity/specificity ratio in ROC curves derived from logistic regression models. RESULTS: 1,636 subjects (51% female and 49% male, mean age 45 [16.3]) were included in the study. Mean plasma homocysteine concentration was 10.7 (4.1) y 8.5 (2.9) micromol/L in men and women, respectively (p < 0.01). Homocysteine levels that best discriminated between cardiovascular disease and gold-standard populations were 10.85 micromol/L in men (sensitivity 58%, specificity 68%), and 9.57 micromol/L in women (sensitivity 50%, specificity 81%). 31.4% of the population (95% CI, 29.1-36.6) presented homocysteine values above these levels. CONCLUSIONS: Hyperhomocysteinemia is not a rare condition in our population. The predictive values obtained imply that measurement of serum homocysteine should be performed only in conjunction with measures of other cardiovascular risk factors. Further research should analyze if homocysteine adds predictive power in cardiovascular risk stratification.

Prevalencia de la hiperhomocisteinemia y factores asociados en atención primaria / Prevalence of hyperhomocysteinemia and associated factors in primary health care

Fundamento y objetivo: Determinar los valores de homocisteína plasmática que pueden considerarse normales en nuestra población y medir la prevalencia de hiperhomocisteinemia. Sujetos y método: Estudio transversal realizado entre todas las personas que acudían (por cualquier motivo) a un centro de salud de Madrid. Se recogió información sociodemográfica, de factores de riesgo, enfermedades y hábitos de vida, y se midieron los valores plasmáticos de homocisteína y otros parámetros bioquímicos. Se calculó el valor de homocisteína que mejor discriminaba a la población con antecedentes de enfermedad vascular de la población de referencia (sujetos sanos con valores plasmáticos normales de vitaminas), mediante análisis de curvas eficacia diagnóstica derivadas de modelos de regresión logística. Resultados: Participaron en el estudio 1.636 personas (un 49% varones y un 51% mujeres), con concentración plasmática media (desviación estándar) de homocisteína de 10,7 (4,1) y 8,5 (2,9) µmol/l, en varones y mujeres respectivamente (p < 0,01). La media de edad fue 45 (16,3) años. El valor de homocisteína que mejor discriminaba el riesgo de presentar enfermedad vascular fue de 10,85 µmol/l en varones (sensibilidad del 58% y especificidad del 68%), y de 9,57 µmol/l en mujeres (sensibilidad del 50% y especificidad del 81%). Un 31,4% de los sujetos (intervalo de confianza del 95%, 29,1-36,6) presentaron concentraciones de homocisteína superiores a estos valores. Conclusiones: La hiperhomocisteinemia es frecuente en nuestra población. El valor predictivo obtenido implica que la determinación de homocisteína sólo debería utilizarse en la valoración de riesgo cardiovascular junto con otros factores de riesgo. Se debe estudiar en el futuro si su determinación añade poder predictivo al estudio de los factores de riesgo clásicos

Background and objective: To identify plasma homocysteine concentrations that could be taken as normal values in our population, and to measure hyperhomocysteinemia prevalence. Subjects and method: Cross-sectional study performed in all subjects attending or working in a primary health care center for any reason. Information was collected about personal history, cardiovascular risk factors and socio-demografic variables, and plasma homocysteine levels and other biochemical parameters were measured. Distribution of homocysteine concentration was compared in individuals with a history of cardiovascular disease and in a gold-standard population (healthy subjects with normal serum concentrations of B-vitamins). The value of homocysteine concentration chosen as cut-off value was the one that offered an optimal sensitivity/specificity ratio in ROC curves derived from logistic regression models. Results: 1,636 subjects (51% female and 49% male, mean age 45 [16.3]) were included in the study. Mean plasma homocysteine concentration was 10.7 (4.1) y 8.5 (2.9) µmol/L in men and women, respectively (p < 0.01). Homocysteine levels that best discriminated between cardiovascular disease and gold-standard populations were 10.85 µmol/L in men (sensitivity 58%, specificity 68%), and 9.57 µmol/L in women (sensitivity 50%, specificity 81%). 31.4% of the population (95% CI, 29.1-36.6) presented homocysteine values above these levels. Conclusions: Hyperhomocysteinemia is not a rare condition in our population. The predictive values obtained imply that measurement of serum homocysteine should be performed only in conjunction with measures of other cardiovascular risk factors. Further research should analyze if homocysteine adds predictive power in cardiovascular risk stratification

[Enzyme replacement therapy in Fabry's disease]. / Terapia de sustitución enzimática en la enfermedad de Fabry: evaluación clínico-histológica tras 18 meses de terapia.

We report a 56-year-old man with history of chronic renal failure, who was diagnosed to have Fabry's disease after performing a percutaneous kidney biopsy. The diagnosis was confirmed by the deficient level of activity of alpha-galactosidase A and by the identification of the mutation. A enzime replacement therapy with alpha-galactosidase A was administered. After 18 months of treatment, a second kidney biopsy was performed showing renal deposits of globotriaosylceramide (we did not evaluate the percentage of histologic clearance of the deposits). Six months after the end of the therapy, a reduction in the impairment of renal function is observed, and the classic manifestations of the disease are absent.

Flow injection spectrophotometric determination of ofloxacin in pharmaceuticals and urine.

A sensitive and fast flow-injection spectrophotometric method for the determination of ofloxacin is proposed. The method is based on the formation of a yellow complex between ofloxacin and Fe (III), in sulphuric medium. The influence of FI and chemical variables were studied. The calibration graph resulting from measuring the absorbance at 420 nm is linear over the range 5.0 x 10(-6)8.0 x 10(-4)M (1.8-289 mg l(-1)) with a detection limit of 2.0 x 10(-6)M (0.72 mg l(-1)). The method is applied to the routine analysis of ofloxacin in pharmaceuticals and human urine.

BCR-ABL induces the expression of Skp2 through the PI3K pathway to promote p27Kip1 degradation and proliferation of chronic myelogenous leukemia cells.

Chronic myelogenous leukemia (CML) is characterized by the expression of the BCR-ABL tyrosine kinase, which results in increased cell proliferation and inhibition of apoptosis. In this study, we show in both BCR-ABL cells (Mo7e-p210 and BaF/3-p210) and primary CML CD34+ cells that STI571 inhibition of BCR-ABL tyrosine kinase activity results in a G(1) cell cycle arrest mediated by the PI3K pathway. This arrest is associated with a nuclear accumulation of p27(Kip1) and down-regulation of cyclins D and E. As a result, there is a reduction of the cyclin E/Cdk2 kinase activity and of the retinoblastoma protein phosphorylation. By quantitative reverse transcription-PCR we show that BCR-ABL/PI3K regulates the expression of p27(Kip1) at the level of transcription. We further show that BCR-ABL also regulates p27(Kip1) protein levels by increasing its degradation by the proteasome. This degradation depends on the ubiquitinylation of p27(Kip1) by Skp2-containing SFC complexes: silencing the expression of Skp2 with a small interfering RNA results in the accumulation of p27(Kip1). We also demonstrate that BCR-ABL cells show transcriptional up-regulation of Skp2. Finally, expression of a p27(Kip1) mutant unable of being recognized by Skp2 results in inhibition of proliferation of BCR-ABL cells, indicating that the degradation of p27(Kip1) contributes to the pathogenesis of CML. In conclusion, these results suggest that BCR-ABL regulates cell cycle in CML cells at least in part by inducing proteasome-mediated degradation of the cell cycle inhibitor p27(Kip1) and provide a rationale for the use of inhibitors of the proteasome in patients with BCR-ABL leukemias.

[Trends in mortality from ischemic heart disease in 50 Spanish provinces]. / Tendencias en la mortalidad por cardiopatía isquémica en 50 provincias españolas.

INTRODUCTION AND OBJECTIVES: Ischemic heart disease is the leading cause of cardiovascular mortality and the second most frequent cause in women in Spain. It is responsible for 12% and 10%, respectively, of all-cause mortality. Our aim was to identify those provinces where recent trends differ significantly from the overall national pattern of decreasing rates in recent years, and where intervention is probably needed. PATIENTS AND METHOD: We report mortality trends due to ischemic heart disease for the periods 1988-1991 and 1994-1997 for each province in persons aged 35 to 64 years, and mortality trends in Spain in the last 19 years for all age groups. Data for ischemic heart disease mortality were obtained from files supplied by the Spanish National Institute for Statistics. Mortality rates in the two 4-year periods were compared with log-linear Poisson regression models to estimate trends. Age-adjusted and mortality hazard-rates are presented with 95% CI, and time trends and geographical variations are also reported. RESULTS: The highest rates were seen at Andalucia (south), Levante (east) and the oversea provinces (Balearic [Mediterranean] and Canary Islands [Atlantic]). There were considerable geographical differences in mortality rates. Time trends showed a decrease in men in 27 provinces and a decrease in women in 12 provinces. Overall mortality trends decreased in the last 19 years in Spain. CONCLUSIONS: Although the general trend is toward a decrease, there are differences between provinces in Spain. The previously reported North-South gradient and regional discrepancies were confirmed.

Tendencias en la mortalidad por cardiopatía isquémica en 50 provincias españolas / Trends in mortality from ischemic heart disease in 50 spanish provinces

Introducción y objetivos. A pesar de la tendencia decreciente de los últimos 25 años, la cardiopatía isquémica continúa siendo la primera causa de muerte en varones y la segunda en mujeres, con un 12 y un 10 por ciento de la mortalidad total, respectivamente. Nuestro objetivo es comprobar si, en el ámbito nacional, el descenso de la mortalidad se mantiene en períodos recientes y en cada una de las provincias, e identificar aquellas donde existen desviaciones significativas del patrón general que aconsejan realizar intervenciones prioritarias. Pacientes y método. Se compara la mortalidad provincial por cardiopatía isquémica en dos cuatrienios (1994-1997 frente a 1988-1991) en la población de 35-64 años, así como la evolución de las tasas de mortalidad en España desde 1980 hasta 1998 para todos los grupos de edad, utilizando datos de defunciones del Instituto Nacional de Estadística. La comparación de la mortalidad entre ambos cuatrienios se ha realizado mediante modelos Poisson de regresión logarítmica lineales, obteniendo la razón de tasas ajustada por edad y sus intervalos de confianza del 95 por ciento. Resultados. Las áreas que presentaron tasas de mortalidad superiores corresponden al sur, levante y regiones insulares. Para todos los grupos de edad, la tendencia en los últimos 19 años es ligeramente decreciente en todo el territorio nacional. Se produce un descenso estadísticamente significativo en 27 provincias entre los varones y en 12 provincias entre las mujeres. En gran parte del territorio, la mortalidad por cardiopatía isquémica no desciende significativamente. Conclusiones. Aunque la tendencia general es decreciente, el descenso no es homogéneo, y se mantienen el patrón norte-sur y las desigualdades territoriales (AU)

Flow-through bulk optode for the fluorimetric determination of perchlorate.

A flow-through fluorescence bulk optode for the flow-injection determination of perchlorate is described. As the active constituents the optode incorporates the lipophilized pH indicator fluorescein octadecyl ester and methyl tridodecyl ammonium chloride, dissolved in a plasticized poly (vinyl) chloride membrane entrapped in a cellulose support. The optode is applied in conjunction with the flow injection technique for perchlorate determination at pH 4.5 (acetic-acetate). The sensor is readily regenerated with the pH 10.4 (TRIS) carrier solution. The analytical characteristics of this optode with respect to perchlorate response time, dynamic measurement range, reproducibility and selectivity are discussed. The proposed FI method is applied to the determination of perchlorate in waters from different sources.

Determination of zinc (II) in pharmaceuticals based on a flow-through bulk optode.

A method based on flow injection (FI), was applied for the determination of Zn (II) using a flow-through bulk optode membrane that incorporates 1-(2-pyridylazo)-2-naphthol in a plasticized poly (vinyl chloride) membrane entrapped in a cellulose support. The calibration graph plotting the reflectance at 562 nm versus [Zn (II)] was linear in the range 0.16-3.27 mgl(-1) (2.5 x 10(-6)-5 x 10(-5) M) with a detection limit of 0.10 mgl(-1). The variation coefficients of the sensor response for 0.33 mgl(-1) of Zn (II) were +/-0.11% for consecutive measurements (n=10), +/-0.19% between days (n=5) and +/-0.22% between different membranes (n=6). The sensor can be readily regenerated with the same acetic/acetate carrier of pH 3.9. The FI method proposed was applied to the determination of zinc (II) in pharmaceuticals.