Performance of Fully-Automated High-Throughput Plasma Biomarker Assays for Alzheimer's Disease in Amnestic Mild Cognitive Impairment Subjects.

INTRODUCTION: Novel plasma biomarkers are promising for identifying Alzheimer's disease (AD) pathological processes in vivo, but most currently employed assays have limitations precluding widespread use. METHODS: CSF and plasma samples were collected from seventy amnestic mild cognitive impairment (aMCI) subjects, stratified as A+ and A-. CSF Aß40, Aß42, p-tau181 and t-tau and plasma Aß40, Aß42 and p-tau181 quantification were conducted using the Lumipulse G assays (Fujirebio), to evaluate the diagnostic performance of plasma biomarkers and assess their associations with CSF biomarkers. RESULTS: All plasma biomarkers except Aß40 showed a very good accuracy in distinguishing A+ aMCI from A- aMCI, Aß42/p-tau181 ratio being the most accurate (AUC 0.895, sensitivity 95.1%, specificity 82.8%). Plasma biomarkers levels were significantly associated with CSF biomarkers concentration. DISCUSSION: High-throughput and fully-automated plasma assays could be helpful in discriminating with high accuracy between aMCI in the AD continuum and aMCI unlikely due to AD in clinical settings.

Effect of calcium deficiency on acyl-CoA synthetase activity in microsomes from various rat tissues

The specific activity of long-chain acyl-CoA synthetase in microsomes from various tissues of control and calcium-deficient rats was determined. It was found that the saturated acids, palmitic and stearic, were preferential substrates compared to the non-saturated linoleic, alpha-linolenic and eicosa-8,11,14-trienoic acids. All of them showed similar Vm values with different affinity constants. After 60-day treatment on a calcium-deficient diet (0.5 g Ca/kg diet), a significant increase in the acyl-CoA synthetase activity was observed for all the tested fatty acids in liver and kidney microsomes. These changes were e voked without any modification in the substrate selectivities shown for the control microsomes, and they were well-correlated with calcium level in both tissues. Under the calcium deficient state an increase in Vm values was observed for palmitic and eicosatrienoic acids with no changes in the corresponding Km, suggesting an increment in the number of active enzyme molecules within the microsomal membrane.

11-deoxycorticosterone, an inducer of a factor stimulating delta 9 desaturase activity in liver microsomes.

The effect of 11-deoxycorticosterone on the oxidative desaturation of [14C]palmitic to palmitoleic acid by liver microsomes was studied in the rat. Twenty-four hours after the hormone injection (1 mumol/kg rat) a significant increase in delta 9 desaturase activity was observed in crude microsomes. A similar stimulation was produced in washed microsomes by addition of a factor present in the 110,000 x g supernatant soluble fraction obtained after extracting crude microsomes from deoxycorticosterone-treated rats with a low ionic strength solution. The deoxycorticosterone-induced factor was also present in the cytosol fraction (110,000 x g supernatant) obtained from livers of hormone-treated animals. Since 11-deoxycorticosterone does not have an 11-beta-OH function, this study demonstrates that such a hydroxyl group is not an absolute requirement for the induction of the soluble factor that regulates delta 9 desaturase activity in response to steroid hormones.

Effect of calcium deficiency on acyl-CoA synthetase activity in microsomes from various rat tissues.

The specific activity of long-chain acyl-CoA synthetase in microsomes from various tissues of control and calcium-deficient rats was determined. It was found that the saturated acids, palmitic and stearic, were preferential substrates compared to the non-saturated linoleic, alpha-linolenic and eicosa-8,11,14-trienoic acids. All of them showed similar Vm values with different affinity constants. After 60-day treatment on a calcium-deficient diet (0.5 g Ca/Kg diet), a significant increase in the acyl-CoA synthetase activity was observed for all the tested fatty acids in liver and kidney microsomes. These changes were evoked without any modification in the substrate selectivities shown for the control microsomes, and they were well-correlated with calcium level in both tissues. Under the calcium deficient state an increase in Vm values was observed for palmitic and eicosatrienoic acids with no changes in the corresponding Km, suggesting an increment in the number of active enzyme molecules within the microsomal membrane.

11-deoxycorticosterone, an inducer of a factor stimulating delta 9 desaturase activity in liver microsomes.

The effect of 11-deoxycorticosterone on the oxidative desaturation of [14C]palmitic to palmitoleic acid by liver microsomes was studied in the rat. Twenty-four hours after the hormone injection (1 mumol/kg rat) a significant increase in delta 9 desaturase activity was observed in crude microsomes. A similar stimulation was produced in washed microsomes by addition of a factor present in the 110,000 x g supernatant soluble fraction obtained after extracting crude microsomes from deoxycorticosterone-treated rats with a low ionic strength solution. The deoxycorticosterone-induced factor was also present in the cytosol fraction (110,000 x g supernatant) obtained from livers of hormone-treated animals. Since 11-deoxycorticosterone does not have an 11-beta-OH function, this study demonstrates that such a hydroxyl group is not an absolute requirement for the induction of the soluble factor that regulates delta 9 desaturase activity in response to steroid hormones.

Effect of calcium deficiency on acyl-CoA synthetase activity in microsomes from various rat tissues

The specific activity of long-chain acyl-CoA synthetase in microsomes from various tissues of control and calcium-deficient rats was determined. It was found that the saturated acids, palmitic and stearic, were preferential substrates compared to the non-saturated linoleic, alpha-linolenic and eicosa-8,11,14-trienoic acids. All of them showed similar Vm values with different affinity constants. After 60-day treatment on a calcium-deficient diet (0.5 g Ca/kg diet), a significant increase in the acyl-CoA synthetase activity was observed for all the tested fatty acids in liver and kidney microsomes. These changes were e voked without any modification in the substrate selectivities shown for the control microsomes, and they were well-correlated with calcium level in both tissues. Under the calcium deficient state an increase in Vm values was observed for palmitic and eicosatrienoic acids with no changes in the corresponding Km, suggesting an increment in the number of active enzyme molecules within the microsomal membrane. (AU)

Hormonal-induced changes on lipidi composition and DPH fluorescence anisotropy of erithrocyte ghost from pre-and postmenopausal women

Fluorescence anisotropy determination were performed on hypotonically lysed red blood cells from 12 healthy, free living, premenopausal and 10 postmenopausal women fed a self-selected diet. Fatty acid composition of phospholipids and cholesterol esters, lipid/lipid and lipid/protein ratios were also determined in erythrocyte ghosts along the menstrual cycle or during hormonal replacemente therapy. The postmenopausal women were treated with transdermal 17-beta estradiol (approx. delivery rate of 50 mug/day) for a 5-week period and with a combination of percutaneous administration of estradiol and orally administered medroxyprogesterone acetate (5 mg/day) for 5 weeks more. Premenopausal women were studied in the mid-follicular and mid-luteal phases of their menstrual cycles. Membrane fluidity in cells from the luteal phase was greater than that observed in the follicular phase. After estradiol treatment, postmenopausal women exhibited a significant raise in the fluorescence anisotropy. The association with progesterone prevented the estrogen-induced increase of the anisotropy values. Estrogen administration also produced an increment in the relative content of cholesterol and membrane proteins that was reverted by progesterone addition. Changes induced by estradiol treatment in the fatty acid pattern of phospholipids and cholesterol esters were characterized by an increment in the monoenoic/saturated fatty acid ratio and linoleic relative content, with a concomitant decrease in the proportion of arachidonic and docosatetraenoic acids. These results suggest that the modification of the desaturase activities may be involved in a mechanism by which these hormonal-induced changes take place. The present study demonstrated that human erythrocyte fluidity is under hormonal control subjected through changes in the proportion and quality of the membrane lipids.

Hormonal-induced changes on lipidi composition and DPH fluorescence anisotropy of erithrocyte ghost from pre-and postmenopausal women

Fluorescence anisotropy determination were performed on hypotonically lysed red blood cells from 12 healthy, free living, premenopausal and 10 postmenopausal women fed a self-selected diet. Fatty acid composition of phospholipids and cholesterol esters, lipid/lipid and lipid/protein ratios were also determined in erythrocyte ghosts along the menstrual cycle or during hormonal replacemente therapy. The postmenopausal women were treated with transdermal 17-beta estradiol (approx. delivery rate of 50 mug/day) for a 5-week period and with a combination of percutaneous administration of estradiol and orally administered medroxyprogesterone acetate (5 mg/day) for 5 weeks more. Premenopausal women were studied in the mid-follicular and mid-luteal phases of their menstrual cycles. Membrane fluidity in cells from the luteal phase was greater than that observed in the follicular phase. After estradiol treatment, postmenopausal women exhibited a significant raise in the fluorescence anisotropy. The association with progesterone prevented the estrogen-induced increase of the anisotropy values. Estrogen administration also produced an increment in the relative content of cholesterol and membrane proteins that was reverted by progesterone addition. Changes induced by estradiol treatment in the fatty acid pattern of phospholipids and cholesterol esters were characterized by an increment in the monoenoic/saturated fatty acid ratio and linoleic relative content, with a concomitant decrease in the proportion of arachidonic and docosatetraenoic acids. These results suggest that the modification of the desaturase activities may be involved in a mechanism by which these hormonal-induced changes take place. The present study demonstrated that human erythrocyte fluidity is under hormonal control subjected through changes in the proportion and quality of the membrane lipids. (AU)

La terapia hormonal sustitutiva modifica la composición en ácidos grasos de fosfolípidos de HDL / Hormone replacement therapy modifies the fatty-acid composition of HDL phospholipids

Las mujeres postmenospáusicas tienen un riesgo aumentado de padecer enfermedad coronaria en comparación con las premenospáusicas. Sin embargo, el riesgo disminuye cuando se realiza terapia hormonal sustitutiva. El objetivo de este estudio es investigar el posible efecto de la terapia con 17 ß Estradiol (E2) o de la combinación de 17 ß Estradiol y Acetato de Medroxiprogesterona (AMP), sobre las concentraciones plásmicas de colesterol total, colesterol HDL, colesterol LDL, colesterol VLDL y triglicéridos, fracciones de conocida participación en la aterogénesis. También se estudió la composición de ácidos grasos de fosfolípidos de la fracción no retenida obtenida por cromatografía de afinidad con Concanavalina A. Al cabo de 30 días de tratamiento con E2, el colesterol total disminuyó desde 226,0 ñ 54,4 mg/dl hasta 202,0 ñ 51,7 mg/dl; los niveles de triglicéridos descendieron desde 106,3 ñ 31,3 mg/dl hasta 80,6 ñ 13,9 mg/dl (p < 0,05), posiblemente a expensas de la fracción VLDL (21,3 ñ 6,2 mg/dl vs. 16,9 ñ 2,5 mg/dl); los fosfolípidos de la fracción no retenida de Concanavalina A mostraron una disminución de los ácidos grasos mirístico, palmítico y esteárico, y un aumento concomitante de los ácidos grasos oleico y linoleico. Los cambios observados con la administración de E2 tendieron a anularse cuando se agregó Acetato de Medroxiprogesterona (AU)

La terapia hormonal sustitutiva modifica la composición en ácidos grasos de fosfolípidos de HDL / Hormone replacement therapy modifies the fatty-acid composition of HDL phospholipids

Las mujeres postmenospáusicas tienen un riesgo aumentado de padecer enfermedad coronaria en comparación con las premenospáusicas. Sin embargo, el riesgo disminuye cuando se realiza terapia hormonal sustitutiva. El objetivo de este estudio es investigar el posible efecto de la terapia con 17 ß Estradiol (E2) o de la combinación de 17 ß Estradiol y Acetato de Medroxiprogesterona (AMP), sobre las concentraciones plásmicas de colesterol total, colesterol HDL, colesterol LDL, colesterol VLDL y triglicéridos, fracciones de conocida participación en la aterogénesis. También se estudió la composición de ácidos grasos de fosfolípidos de la fracción no retenida obtenida por cromatografía de afinidad con Concanavalina A. Al cabo de 30 días de tratamiento con E2, el colesterol total disminuyó desde 226,0 ñ 54,4 mg/dl hasta 202,0 ñ 51,7 mg/dl; los niveles de triglicéridos descendieron desde 106,3 ñ 31,3 mg/dl hasta 80,6 ñ 13,9 mg/dl (p < 0,05), posiblemente a expensas de la fracción VLDL (21,3 ñ 6,2 mg/dl vs. 16,9 ñ 2,5 mg/dl); los fosfolípidos de la fracción no retenida de Concanavalina A mostraron una disminución de los ácidos grasos mirístico, palmítico y esteárico, y un aumento concomitante de los ácidos grasos oleico y linoleico. Los cambios observados con la administración de E2 tendieron a anularse cuando se agregó Acetato de Medroxiprogesterona