Francisco Blanco-vaca - Academia.edu (original) (raw)

Papers by Francisco Blanco-vaca

Medicina Clínica, 2005

La aproximación bioquímica clásica en el diagnóstico y, sobre todo, el control del cáncer se basa... more La aproximación bioquímica clásica en el diagnóstico y, sobre todo, el control del cáncer se basa en la determinación de la concentración en suero de determinadas proteínas producidas por el tejido tumoral, denominadas marcadores tumorales. Los métodos utilizados requieren el uso de anticuerpos específicos, por lo que suelen englobarse bajo el término genérico de inmunoanálisis. Estas técnicas detectan proteínas individuales (p. ej., antígeno prostático específico [PSA]) en concentraciones muy bajas (µg/l). El uso de marcadores tumorales para el diagnóstico de cáncer presenta conocidas limitaciones de sensibilidad y especificidad. Así, las enfermedades benignas relacionadas con el tejido u órgano en estudio o con el órgano de eliminación del marcador (patología hepática o renal, principalmente) aumentan su concentración en el suero y disminuyen la especificidad diagnóstica. La sensibilidad diagnóstica de los marcadores tumorales tampoco es del 100% y está relacionada con el estadio del tumor: es baja en los estadios iniciales y aumenta con la progresión tumoral. Estas limitaciones hacen que, en muchos casos, la utilidad de los marcadores tumorales durante el proceso de diagnóstico del cáncer sea limitada y que, en cambio, su mayor utilidad se encuentre en el seguimiento de la enfermedad y en la valoración de la respuesta al tratamiento.

L'estudi del metabolisme de les HDL (lipoproteïnes de densitat alta) i la seva relació amb l&... more L'estudi del metabolisme de les HDL (lipoproteïnes de densitat alta) i la seva relació amb l'arteriosclerosi és un tema complex, en el qual, recentment, s'han fet avenços importants. Primer, s'ha demostrat que les HDL tenen una acció antiaterogènica. Segon, s'ha establert que aquesta acció és deguda a fraccions que tenen apoA-I però no apoA-II. Tercer, s'han definit alguns mecanismes clau

Clinica Chimica Acta, 2016

The adequate absorption of lipids is essential for all mammalian species due to their inability t... more The adequate absorption of lipids is essential for all mammalian species due to their inability to synthesize some essential fatty acids and fat-soluble vitamins. Chylomicrons (CMs) are large, triglyceride-rich lipoproteins that are produced in intestinal enterocytes in response to fat ingestion, which function to transport the ingested lipids to different tissues. In addition to the contribution of CMs to postprandial lipemia, their remnants, the degradation products following lipolysis by lipoprotein lipase, are linked to cardiovascular disease. In this review, we will focus on the structure-function and metabolism of CMs. Second, we will analyze the impact of gene defects reported to affect CM metabolism and, also, the role of CMs in other pathologies, such as atherothrombotic cardiovascular disease and diabetes mellitus. Third, we will provide an overview of the laboratory tests currently used to study CM disorders, and, finally, we will highlight current treatments in diseases affecting CMs.

Haematologica

Recent studies suggest that mild hyperhomocysteinemia may be a risk factor for venous thromboembo... more Recent studies suggest that mild hyperhomocysteinemia may be a risk factor for venous thromboembolic disease (VTED). In this work we evaluated the prevalence of moderate hyperhomocysteinemia in patients with VTED in our area. We found hyperhomocysteinemia in 23.4% of 64 patients studied compared with 7.35% of 68 healthy controls (p=0.014). Our results suggest that moderate hyperhomocysteinemia is one of the most prevalent abnormalities associated with VTED.

Methods in Molecular Biology, 2015

Promotion of reverse cholesterol transport (RCT) is thought to be a major HDL-mediated mechanism ... more Promotion of reverse cholesterol transport (RCT) is thought to be a major HDL-mediated mechanism for protecting against atherosclerosis. Preclinical studies support the concept that increasing cholesterol efflux from macrophages may confer atheroprotective benefits independently of the plasma HDL-cholesterol concentration. The application of the macrophage-to-feces RCT method in genetically engineered mice has provided evidence that this major HDL property correlates closely with changes in atherosclerosis susceptibility. This chapter provides details on the methodologies currently used to measure in vitro cholesterol efflux from macrophages or in vivo macrophage-specific RCT. The general principles and techniques described herein may be applied to measure the in vitro cholesterol efflux capacity of human serum in macrophage cultures and to evaluate the effect of different experimental pathophysiological conditions or the efficacy of different therapeutic strategies on the modulation of in vivo macrophage-RCT in mice.

The Journal of Lipid Research

Familial combined hyperlipidemia (FCHL) is a common inherited hyperlipidemia and a major risk fac... more Familial combined hyperlipidemia (FCHL) is a common inherited hyperlipidemia and a major risk factor for atherothrombotic cardiovascular disease. The cause(s) leading to FCHL are largely unknown, but the existence of unidentified "major" genes that would increase VLDL production and of "modifier" genes that would influence the phenotype of the disease has been proposed. Expression of apolipoprotein A-II (apoA-II), a high density lipoprotein (HDL) of unknown function, in transgenic mice produced increased concentration of apoB-containing lipoproteins and decreased HDL. Here we show that expression of human apoA-II in apoE-deficient mice induces a dose-dependent increase in VLDL, resulting in plasma triglyceride elevations of up to 24-fold in a mouse line that has 2-fold the concentration of human apoA-II of normolipidemic humans, as well as other well-known characteristics of FCHL: increased concentrations of cholesterol, triglyceride, and apoB in very low density...

Journal of lipid research, 2001

Apolipoprotein (apo)A-II is a major high density lipoprotein (HDL) protein; however, its role in ... more Apolipoprotein (apo)A-II is a major high density lipoprotein (HDL) protein; however, its role in lipoprotein metabolism is largely unknown. Transgenic (Tg) mice that overexpress human apoA-II present functional lecithin: cholesterol acyltransferase deficiency, HDL deficiency, hypertriglyceridemia and, when fed an atherogenic diet, increased non-HDL cholesterol and increased susceptibility to atherosclerosis. In contrast to humans, mice do not present cholesteryl ester transfer protein (CETP) activity in plasma. To study the in vivo interaction of these two proteins, we crossbred human apoA-II and CETP-Tg mice. CETP x apoA-II-Tg mice fed an atherogenic diet, compared with CETP-Tg mice presented a 2-fold decrease in HDL cholesterol and a quantitatively similar increase in total plasma cholesterol and percentage of free cholesterol, non-HDL cholesterol, and free fatty acids, together with a remarkable 112-fold increase in plasma triglycerides. Plasma triglycerides in CETP x apoA-II-Tg ...

Clinical chemistry, 2000

LDL-cholesterol (LDLc) is the main lipid marker in cardiovascular risk estimation and the princip... more LDL-cholesterol (LDLc) is the main lipid marker in cardiovascular risk estimation and the principal therapeutic target in both diabetic and nondiabetic subjects (1, 2 ). The designated comparison method for the determination of LDLc, using ultracentrifugation and precipitation, known as "␤-quantification" (3 ), is cumbersome and time-consuming and requires expensive instrumentation and trained personnel. The Friedewald equation (4 ) {LDLc ϭ total cholesterol Ϫ HDLc Ϫ [triglycerides (in mmol/L)/ 2.17 or triglycerides (in mg/dL)/5]}, the most frequently used method for the calculation of LDLc, assumes that VLDL particles maintain a nearly constant cholesterol: triglyceride ratio. However, this assumption is invalid in the presence of chylomicronemia and increased VLDL or intermediate-density lipoprotein particles (4 -7 ).

Journal of lipid research, 1995

Fish-eye disease (FED) and familial lecithin:cholesterol acyltransferase (LCAT) deficiency (FLD) ... more Fish-eye disease (FED) and familial lecithin:cholesterol acyltransferase (LCAT) deficiency (FLD) are rare disorders of lipid metabolism linked to mutations in the LCAT gene. Eleven LCAT cDNA constructs associated with FED and FLD were prepared by site-directed mutagenesis and expressed in COS-6 cells. Analysis of total RNA from wild-type, FED, and FLD transfectants revealed that all contained LCAT-specific mRNA. Western blot analysis demonstrated that all LCAT transfectants synthesized LCAT. Mean LCAT secretion by FED transfectants was slightly lower than secretion by wild-type transfectants, whereas secretion by FLD transfectants was much lower. The specific activities of FED and FLD LCAT against model high density lipoproteins were 6% and 11%, respectively, of wild-type activity. The ratios of the LCAT activities against low density lipoproteins to those against model high density lipoproteins decreased in the order FED mutants > FLD mutants approximately wild type. FED and FLD...

Journal of lipid research, 2001

Our understanding of apolipoprotein A-II (apoA-II) physiology is much more limited than that of a... more Our understanding of apolipoprotein A-II (apoA-II) physiology is much more limited than that of apoA-I. However, important and rather surprising advances have been produced, mainly through analysis of genetically modified mice. These results reveal a positive association of apoA-II with FFA and VLDL triglyceride plasma concentrations; however, whether this is due to increased VLDL synthesis or to decreased VLDL catabolism remains a matter of controversy. As apoA-II-deficient mice present a phenotype of insulin hypersensitivity, a function of apoA-II in regulating FFA metabolism seems likely. Studies of human beings have shown the apoA-II locus to be a determinant of FFA plasma levels, and several genome-wide searches of different populations with type 2 diabetes have found linkage to an apoA-II intragenic marker, making apoA-II an attractive candidate gene for this disease. The increased concentration of apoB-containing lipoproteins present in apoA-II transgenic mice explains, in pa...

The Journal of Lipid Research

The density distribution of electronegative LDL [LDL(-)], a cytotoxic and inflammatory fraction o... more The density distribution of electronegative LDL [LDL(-)], a cytotoxic and inflammatory fraction of LDL present in plasma, was studied in 10 normolipemic (NL), 6 FH, and 11 hypertriglyceridemic (HTG) subjects. Six LDL subclasses of increased density (LDL1 to LDL6) were isolated by density-gradient ultracentrifugation (DGU). NL and FH subjects showed prevalence of light LDL, whereas HTG subjects showed prevalence of dense LDL. LDL(-) proportion was determined from total LDL or LDL-density subclasses by anion-exchange chromatography. LDL from FH patients had increased LDL(-) (35.1 +/- 9.9%) compared with LDL from NL and HTG subjects (9.4 +/- 2.3% and 12.3 +/- 4.3%, respectively). Most LDL(-) was contained in dense subclasses in NL (LDL4-6, 67.7 +/- 3.1%) whereas most of LDL(-) from FH patients were contained in light LDL subclasses (LDL1-3) (86.2 +/- 1.6%). In these subjects, simvastatin therapy decreased LDL(-) to 28.2 +/- 6.7% and 21.2 +/- 5.6% at 3 and 6 months of treatment, respect...

International journal of biomedical science : IJBS, 2009

A method to evaluate the antioxidant capacity of high-density lipoprotein (HDL) was developed and... more A method to evaluate the antioxidant capacity of high-density lipoprotein (HDL) was developed and standardized. This method measure conjugated diene (CD) formation and electrophoretic mobility of low-density lipoprotein (LDL) in agarose gels in the presence and absence of HDL. HDL was isolated from 1 mL of plasma within 24 hours and oxidation assays were performed within 6 hours. Oxidation was induced by adding CuSO4. The lag phase increase in CD kinetics and the inhibition of electrophoretic mobility were defined as the HDL antioxidant capacity. The optimal conditions for the CD assay were 2.5 μM CuSO4, LDL at 0.1 g apoB/L, HDL at 0.1 g apoA-I/L, at 37°C and for 3h 50 min. Agarose electrophoresis at 100 V, at 4°C for 50 min was then performed immediately. CD formation variability was 21.1% for inter-assay CV and 12.7% for intra-assay CV. Electrophoretic mobility was 26.5% for inter-assay CV and 2.4% for intra-assay CV. Correlation analysis showed a significant association between t...

Clínica e Investigación en Arteriosclerosis, 2013

Recibido el 7 de mayo de 2013; aceptado el 17 de mayo de 2013 Disponible en Internet el 12 de jun... more Recibido el 7 de mayo de 2013; aceptado el 17 de mayo de 2013 Disponible en Internet el 12 de junio de 2013

Journal of lipid research, 2002

Apolipoprotein (apo)A-I is the major protein component of HDL and the cofactor for LCAT. We descr... more Apolipoprotein (apo)A-I is the major protein component of HDL and the cofactor for LCAT. We describe a large Spanish kindred, living in the Mediterranean Island of Mallorca, that presents a dominant form of hypoalphalipoproteinemia. The lipid profile of this family was studied because the proband, a 40-year-old male presenting signs of coronary atherosclerosis, showed severe HDL deficiency. However, none of the other family members had a known history of cardiovascular disease. Sequence analysis of the apoA-I gene in affected members identified a 33-base pair deletion, corresponding to residues 165-175 of the mature protein, eliminating the first 11 amino acids of the internal repeat 7. ApoA-I(MALLORCA) is associated with HDL-cholesterol deficiency (concentration ranging from 8-48% of the value in non-carriers), and a 2- to 3-fold decrease in plasma concentrations of apoA-I and apoA-II and endogenous LCAT activity, concomitant with a slight decrease in serum cholesterol efflux capab...

Clínica e Investigación en Arteriosclerosis, 2010

Recibido el 19 de mayo de 2010; aceptado el 4 de junio de 2010 Disponible en Internet el 18 de se... more Recibido el 19 de mayo de 2010; aceptado el 4 de junio de 2010 Disponible en Internet el 18 de septiembre de 2010 PALABRAS CLAVE ApoA-II; ApoC; HDL; Lipoproteína lipasa; Triglicéridos

Nutrition journal, Jan 15, 2004

Although the consumption of wine in particular has been associated with a lower risk of atherothr... more Although the consumption of wine in particular has been associated with a lower risk of atherothrombotic cardiovascular disease, systematic reviews differ as to the relative protective effect of beer, wine and spirits. Two previous studies showed that red wine reduces fatty streak formation (early atherosclerosis) but not mature atherosclerosis in apolipoprotein (apo) E-deficient (apoE-/-) mice. To determine whether a moderate beer intake would affect early and mature atherosclerotic lesion formation using control C57BL/6 and apoE-/- mice, respectively, as models. Control C57BL/6 and apoE-/- mice were randomized to receive either water, ethanol, mild beer, dark beer or ethanol-free beer. The level of beer was designed to approximate the alcohol intake currently believed to be beneficial in reducing human vascular risk. Control C57BL/6 mice were fed a Western diet for 24 weeks, and apoE-/- mice a chow diet for 12 weeks. At the end of the trial period, mice were euthanized and atheros...

Molecular Nutrition & Food Research, 2013

Scope: High plasma homocysteine concentrations have been associated with increased risk of cardio... more Scope: High plasma homocysteine concentrations have been associated with increased risk of cardiovascular disease both in humans and experimental animal models, whereas plasma HDL-cholesterol concentration is inversely correlated with such disorders. This work aimed to study the impact of methionine-induced hyperhomocysteinemia (HHcy) on two major antiatherogenic functions of HDL, namely their capacity to prevent LDL oxidation and induce in vivo macrophage-specific reverse cholesterol transport. Methods and results: Methionine-induced HHcy in mice resulted in an approximately 20% decreased concentration of HDL-cholesterol and HDL main protein component, apolipoprotein A-I. The HDL potential to resist oxidation as well as to prevent LDL oxidative modification was impaired in hyperhomocysteinemic mice. Activities of paraoxonase-1 and platelet activation factor acetylhydrolase, two of the main HDL-associated enzymes with antioxidant activity, were reduced. The ability of HDL to efflux cholesterol from macrophages was decreased in hyperhomocysteinemic mice; however, the in vivo macrophage-specific reverse cholesterol transport measured as the output of labeled cholesterol into feces did not significantly differ between groups. Conclusion: Our data indicate that the HDL from methionine-induced hyperhomocysteinemic mice was more prone to oxidation and displayed lower capacity to protect LDL against oxidative modification than that of control mice, highlighting a mechanism by which a diet-induced HHcy may facilitate progression of atherosclerosis.

Revista Clínica Española

In this paper we present the results of a study about the diagnostic utility of Adenosine Deamina... more In this paper we present the results of a study about the diagnostic utility of Adenosine Deaminase (ADA) determination for diagnosing the tuberculous pleural effusion. We carried out this study in 71 patients who came to our Hospital and were diagnosed for pleural effusion. The ADA determination was made in sera and pleural effusion of all these patients. Also we determined how much of this ADA activity was linked to microsomes or not (soluble ADA), by ultracentrifugation as a separative method. Taking as a base the data obtained and using 43 U/l as cutoff point, the Diagnostic Sensitivity of ADA determined in pleural liquid calculated was 1. This means that all patients with tuberculous pleural effusions had ADA activities higher than 43 U/l in their pleural liquid. On the other hand, Diagnostic Specificity was of 0.83. This was due to the existence of 10 false positives: 2 malignancies, 3 empyemas, 1 postpneumonia, 1 secondary to thromboembolic pulmonary disease, 1 secondary to rheumatoid arthritis and 2 of unknown origin. Results obtained show the utility, with some limitations, of ADA determination in pleural liquid in order to diagnose tuberculous pleural effusions. Neither ADA determination in sera nor quantity of ADA linked to microsomes or solubles ADA in pleural liquid seem to have great clinical interest.

Medicina Clínica, 2005

La aproximación bioquímica clásica en el diagnóstico y, sobre todo, el control del cáncer se basa... more La aproximación bioquímica clásica en el diagnóstico y, sobre todo, el control del cáncer se basa en la determinación de la concentración en suero de determinadas proteínas producidas por el tejido tumoral, denominadas marcadores tumorales. Los métodos utilizados requieren el uso de anticuerpos específicos, por lo que suelen englobarse bajo el término genérico de inmunoanálisis. Estas técnicas detectan proteínas individuales (p. ej., antígeno prostático específico [PSA]) en concentraciones muy bajas (µg/l). El uso de marcadores tumorales para el diagnóstico de cáncer presenta conocidas limitaciones de sensibilidad y especificidad. Así, las enfermedades benignas relacionadas con el tejido u órgano en estudio o con el órgano de eliminación del marcador (patología hepática o renal, principalmente) aumentan su concentración en el suero y disminuyen la especificidad diagnóstica. La sensibilidad diagnóstica de los marcadores tumorales tampoco es del 100% y está relacionada con el estadio del tumor: es baja en los estadios iniciales y aumenta con la progresión tumoral. Estas limitaciones hacen que, en muchos casos, la utilidad de los marcadores tumorales durante el proceso de diagnóstico del cáncer sea limitada y que, en cambio, su mayor utilidad se encuentre en el seguimiento de la enfermedad y en la valoración de la respuesta al tratamiento.

L'estudi del metabolisme de les HDL (lipoproteïnes de densitat alta) i la seva relació amb l&... more L'estudi del metabolisme de les HDL (lipoproteïnes de densitat alta) i la seva relació amb l'arteriosclerosi és un tema complex, en el qual, recentment, s'han fet avenços importants. Primer, s'ha demostrat que les HDL tenen una acció antiaterogènica. Segon, s'ha establert que aquesta acció és deguda a fraccions que tenen apoA-I però no apoA-II. Tercer, s'han definit alguns mecanismes clau

Clinica Chimica Acta, 2016

The adequate absorption of lipids is essential for all mammalian species due to their inability t... more The adequate absorption of lipids is essential for all mammalian species due to their inability to synthesize some essential fatty acids and fat-soluble vitamins. Chylomicrons (CMs) are large, triglyceride-rich lipoproteins that are produced in intestinal enterocytes in response to fat ingestion, which function to transport the ingested lipids to different tissues. In addition to the contribution of CMs to postprandial lipemia, their remnants, the degradation products following lipolysis by lipoprotein lipase, are linked to cardiovascular disease. In this review, we will focus on the structure-function and metabolism of CMs. Second, we will analyze the impact of gene defects reported to affect CM metabolism and, also, the role of CMs in other pathologies, such as atherothrombotic cardiovascular disease and diabetes mellitus. Third, we will provide an overview of the laboratory tests currently used to study CM disorders, and, finally, we will highlight current treatments in diseases affecting CMs.

Haematologica

Recent studies suggest that mild hyperhomocysteinemia may be a risk factor for venous thromboembo... more Recent studies suggest that mild hyperhomocysteinemia may be a risk factor for venous thromboembolic disease (VTED). In this work we evaluated the prevalence of moderate hyperhomocysteinemia in patients with VTED in our area. We found hyperhomocysteinemia in 23.4% of 64 patients studied compared with 7.35% of 68 healthy controls (p=0.014). Our results suggest that moderate hyperhomocysteinemia is one of the most prevalent abnormalities associated with VTED.

Methods in Molecular Biology, 2015

Promotion of reverse cholesterol transport (RCT) is thought to be a major HDL-mediated mechanism ... more Promotion of reverse cholesterol transport (RCT) is thought to be a major HDL-mediated mechanism for protecting against atherosclerosis. Preclinical studies support the concept that increasing cholesterol efflux from macrophages may confer atheroprotective benefits independently of the plasma HDL-cholesterol concentration. The application of the macrophage-to-feces RCT method in genetically engineered mice has provided evidence that this major HDL property correlates closely with changes in atherosclerosis susceptibility. This chapter provides details on the methodologies currently used to measure in vitro cholesterol efflux from macrophages or in vivo macrophage-specific RCT. The general principles and techniques described herein may be applied to measure the in vitro cholesterol efflux capacity of human serum in macrophage cultures and to evaluate the effect of different experimental pathophysiological conditions or the efficacy of different therapeutic strategies on the modulation of in vivo macrophage-RCT in mice.

The Journal of Lipid Research

Familial combined hyperlipidemia (FCHL) is a common inherited hyperlipidemia and a major risk fac... more Familial combined hyperlipidemia (FCHL) is a common inherited hyperlipidemia and a major risk factor for atherothrombotic cardiovascular disease. The cause(s) leading to FCHL are largely unknown, but the existence of unidentified "major" genes that would increase VLDL production and of "modifier" genes that would influence the phenotype of the disease has been proposed. Expression of apolipoprotein A-II (apoA-II), a high density lipoprotein (HDL) of unknown function, in transgenic mice produced increased concentration of apoB-containing lipoproteins and decreased HDL. Here we show that expression of human apoA-II in apoE-deficient mice induces a dose-dependent increase in VLDL, resulting in plasma triglyceride elevations of up to 24-fold in a mouse line that has 2-fold the concentration of human apoA-II of normolipidemic humans, as well as other well-known characteristics of FCHL: increased concentrations of cholesterol, triglyceride, and apoB in very low density...

Journal of lipid research, 2001

Apolipoprotein (apo)A-II is a major high density lipoprotein (HDL) protein; however, its role in ... more Apolipoprotein (apo)A-II is a major high density lipoprotein (HDL) protein; however, its role in lipoprotein metabolism is largely unknown. Transgenic (Tg) mice that overexpress human apoA-II present functional lecithin: cholesterol acyltransferase deficiency, HDL deficiency, hypertriglyceridemia and, when fed an atherogenic diet, increased non-HDL cholesterol and increased susceptibility to atherosclerosis. In contrast to humans, mice do not present cholesteryl ester transfer protein (CETP) activity in plasma. To study the in vivo interaction of these two proteins, we crossbred human apoA-II and CETP-Tg mice. CETP x apoA-II-Tg mice fed an atherogenic diet, compared with CETP-Tg mice presented a 2-fold decrease in HDL cholesterol and a quantitatively similar increase in total plasma cholesterol and percentage of free cholesterol, non-HDL cholesterol, and free fatty acids, together with a remarkable 112-fold increase in plasma triglycerides. Plasma triglycerides in CETP x apoA-II-Tg ...

Clinical chemistry, 2000

LDL-cholesterol (LDLc) is the main lipid marker in cardiovascular risk estimation and the princip... more LDL-cholesterol (LDLc) is the main lipid marker in cardiovascular risk estimation and the principal therapeutic target in both diabetic and nondiabetic subjects (1, 2 ). The designated comparison method for the determination of LDLc, using ultracentrifugation and precipitation, known as "␤-quantification" (3 ), is cumbersome and time-consuming and requires expensive instrumentation and trained personnel. The Friedewald equation (4 ) {LDLc ϭ total cholesterol Ϫ HDLc Ϫ [triglycerides (in mmol/L)/ 2.17 or triglycerides (in mg/dL)/5]}, the most frequently used method for the calculation of LDLc, assumes that VLDL particles maintain a nearly constant cholesterol: triglyceride ratio. However, this assumption is invalid in the presence of chylomicronemia and increased VLDL or intermediate-density lipoprotein particles (4 -7 ).

Journal of lipid research, 1995

Fish-eye disease (FED) and familial lecithin:cholesterol acyltransferase (LCAT) deficiency (FLD) ... more Fish-eye disease (FED) and familial lecithin:cholesterol acyltransferase (LCAT) deficiency (FLD) are rare disorders of lipid metabolism linked to mutations in the LCAT gene. Eleven LCAT cDNA constructs associated with FED and FLD were prepared by site-directed mutagenesis and expressed in COS-6 cells. Analysis of total RNA from wild-type, FED, and FLD transfectants revealed that all contained LCAT-specific mRNA. Western blot analysis demonstrated that all LCAT transfectants synthesized LCAT. Mean LCAT secretion by FED transfectants was slightly lower than secretion by wild-type transfectants, whereas secretion by FLD transfectants was much lower. The specific activities of FED and FLD LCAT against model high density lipoproteins were 6% and 11%, respectively, of wild-type activity. The ratios of the LCAT activities against low density lipoproteins to those against model high density lipoproteins decreased in the order FED mutants > FLD mutants approximately wild type. FED and FLD...

Journal of lipid research, 2001

Our understanding of apolipoprotein A-II (apoA-II) physiology is much more limited than that of a... more Our understanding of apolipoprotein A-II (apoA-II) physiology is much more limited than that of apoA-I. However, important and rather surprising advances have been produced, mainly through analysis of genetically modified mice. These results reveal a positive association of apoA-II with FFA and VLDL triglyceride plasma concentrations; however, whether this is due to increased VLDL synthesis or to decreased VLDL catabolism remains a matter of controversy. As apoA-II-deficient mice present a phenotype of insulin hypersensitivity, a function of apoA-II in regulating FFA metabolism seems likely. Studies of human beings have shown the apoA-II locus to be a determinant of FFA plasma levels, and several genome-wide searches of different populations with type 2 diabetes have found linkage to an apoA-II intragenic marker, making apoA-II an attractive candidate gene for this disease. The increased concentration of apoB-containing lipoproteins present in apoA-II transgenic mice explains, in pa...

The Journal of Lipid Research

The density distribution of electronegative LDL [LDL(-)], a cytotoxic and inflammatory fraction o... more The density distribution of electronegative LDL [LDL(-)], a cytotoxic and inflammatory fraction of LDL present in plasma, was studied in 10 normolipemic (NL), 6 FH, and 11 hypertriglyceridemic (HTG) subjects. Six LDL subclasses of increased density (LDL1 to LDL6) were isolated by density-gradient ultracentrifugation (DGU). NL and FH subjects showed prevalence of light LDL, whereas HTG subjects showed prevalence of dense LDL. LDL(-) proportion was determined from total LDL or LDL-density subclasses by anion-exchange chromatography. LDL from FH patients had increased LDL(-) (35.1 +/- 9.9%) compared with LDL from NL and HTG subjects (9.4 +/- 2.3% and 12.3 +/- 4.3%, respectively). Most LDL(-) was contained in dense subclasses in NL (LDL4-6, 67.7 +/- 3.1%) whereas most of LDL(-) from FH patients were contained in light LDL subclasses (LDL1-3) (86.2 +/- 1.6%). In these subjects, simvastatin therapy decreased LDL(-) to 28.2 +/- 6.7% and 21.2 +/- 5.6% at 3 and 6 months of treatment, respect...

International journal of biomedical science : IJBS, 2009

A method to evaluate the antioxidant capacity of high-density lipoprotein (HDL) was developed and... more A method to evaluate the antioxidant capacity of high-density lipoprotein (HDL) was developed and standardized. This method measure conjugated diene (CD) formation and electrophoretic mobility of low-density lipoprotein (LDL) in agarose gels in the presence and absence of HDL. HDL was isolated from 1 mL of plasma within 24 hours and oxidation assays were performed within 6 hours. Oxidation was induced by adding CuSO4. The lag phase increase in CD kinetics and the inhibition of electrophoretic mobility were defined as the HDL antioxidant capacity. The optimal conditions for the CD assay were 2.5 μM CuSO4, LDL at 0.1 g apoB/L, HDL at 0.1 g apoA-I/L, at 37°C and for 3h 50 min. Agarose electrophoresis at 100 V, at 4°C for 50 min was then performed immediately. CD formation variability was 21.1% for inter-assay CV and 12.7% for intra-assay CV. Electrophoretic mobility was 26.5% for inter-assay CV and 2.4% for intra-assay CV. Correlation analysis showed a significant association between t...

Clínica e Investigación en Arteriosclerosis, 2013

Recibido el 7 de mayo de 2013; aceptado el 17 de mayo de 2013 Disponible en Internet el 12 de jun... more Recibido el 7 de mayo de 2013; aceptado el 17 de mayo de 2013 Disponible en Internet el 12 de junio de 2013

Journal of lipid research, 2002

Apolipoprotein (apo)A-I is the major protein component of HDL and the cofactor for LCAT. We descr... more Apolipoprotein (apo)A-I is the major protein component of HDL and the cofactor for LCAT. We describe a large Spanish kindred, living in the Mediterranean Island of Mallorca, that presents a dominant form of hypoalphalipoproteinemia. The lipid profile of this family was studied because the proband, a 40-year-old male presenting signs of coronary atherosclerosis, showed severe HDL deficiency. However, none of the other family members had a known history of cardiovascular disease. Sequence analysis of the apoA-I gene in affected members identified a 33-base pair deletion, corresponding to residues 165-175 of the mature protein, eliminating the first 11 amino acids of the internal repeat 7. ApoA-I(MALLORCA) is associated with HDL-cholesterol deficiency (concentration ranging from 8-48% of the value in non-carriers), and a 2- to 3-fold decrease in plasma concentrations of apoA-I and apoA-II and endogenous LCAT activity, concomitant with a slight decrease in serum cholesterol efflux capab...

Clínica e Investigación en Arteriosclerosis, 2010

Recibido el 19 de mayo de 2010; aceptado el 4 de junio de 2010 Disponible en Internet el 18 de se... more Recibido el 19 de mayo de 2010; aceptado el 4 de junio de 2010 Disponible en Internet el 18 de septiembre de 2010 PALABRAS CLAVE ApoA-II; ApoC; HDL; Lipoproteína lipasa; Triglicéridos

Nutrition journal, Jan 15, 2004

Although the consumption of wine in particular has been associated with a lower risk of atherothr... more Although the consumption of wine in particular has been associated with a lower risk of atherothrombotic cardiovascular disease, systematic reviews differ as to the relative protective effect of beer, wine and spirits. Two previous studies showed that red wine reduces fatty streak formation (early atherosclerosis) but not mature atherosclerosis in apolipoprotein (apo) E-deficient (apoE-/-) mice. To determine whether a moderate beer intake would affect early and mature atherosclerotic lesion formation using control C57BL/6 and apoE-/- mice, respectively, as models. Control C57BL/6 and apoE-/- mice were randomized to receive either water, ethanol, mild beer, dark beer or ethanol-free beer. The level of beer was designed to approximate the alcohol intake currently believed to be beneficial in reducing human vascular risk. Control C57BL/6 mice were fed a Western diet for 24 weeks, and apoE-/- mice a chow diet for 12 weeks. At the end of the trial period, mice were euthanized and atheros...

Molecular Nutrition & Food Research, 2013

Scope: High plasma homocysteine concentrations have been associated with increased risk of cardio... more Scope: High plasma homocysteine concentrations have been associated with increased risk of cardiovascular disease both in humans and experimental animal models, whereas plasma HDL-cholesterol concentration is inversely correlated with such disorders. This work aimed to study the impact of methionine-induced hyperhomocysteinemia (HHcy) on two major antiatherogenic functions of HDL, namely their capacity to prevent LDL oxidation and induce in vivo macrophage-specific reverse cholesterol transport. Methods and results: Methionine-induced HHcy in mice resulted in an approximately 20% decreased concentration of HDL-cholesterol and HDL main protein component, apolipoprotein A-I. The HDL potential to resist oxidation as well as to prevent LDL oxidative modification was impaired in hyperhomocysteinemic mice. Activities of paraoxonase-1 and platelet activation factor acetylhydrolase, two of the main HDL-associated enzymes with antioxidant activity, were reduced. The ability of HDL to efflux cholesterol from macrophages was decreased in hyperhomocysteinemic mice; however, the in vivo macrophage-specific reverse cholesterol transport measured as the output of labeled cholesterol into feces did not significantly differ between groups. Conclusion: Our data indicate that the HDL from methionine-induced hyperhomocysteinemic mice was more prone to oxidation and displayed lower capacity to protect LDL against oxidative modification than that of control mice, highlighting a mechanism by which a diet-induced HHcy may facilitate progression of atherosclerosis.

Revista Clínica Española

In this paper we present the results of a study about the diagnostic utility of Adenosine Deamina... more In this paper we present the results of a study about the diagnostic utility of Adenosine Deaminase (ADA) determination for diagnosing the tuberculous pleural effusion. We carried out this study in 71 patients who came to our Hospital and were diagnosed for pleural effusion. The ADA determination was made in sera and pleural effusion of all these patients. Also we determined how much of this ADA activity was linked to microsomes or not (soluble ADA), by ultracentrifugation as a separative method. Taking as a base the data obtained and using 43 U/l as cutoff point, the Diagnostic Sensitivity of ADA determined in pleural liquid calculated was 1. This means that all patients with tuberculous pleural effusions had ADA activities higher than 43 U/l in their pleural liquid. On the other hand, Diagnostic Specificity was of 0.83. This was due to the existence of 10 false positives: 2 malignancies, 3 empyemas, 1 postpneumonia, 1 secondary to thromboembolic pulmonary disease, 1 secondary to rheumatoid arthritis and 2 of unknown origin. Results obtained show the utility, with some limitations, of ADA determination in pleural liquid in order to diagnose tuberculous pleural effusions. Neither ADA determination in sera nor quantity of ADA linked to microsomes or solubles ADA in pleural liquid seem to have great clinical interest.