Apolipoprotein C-III protein concentrations and gene polymorphisms in Type 1 diabetes:associations with microvascular disease complications in the DCCT/EDIC cohort

We investigated the associations of apolipoprotein C-III (apoCIII) protein and apoCIII gene variation with microvascular disease complications in Type 1 diabetes.

Apolipoprotein C-III protein concentrations and gene polymorphisms in type 1 diabetes:associations with lipoprotein subclasses

Serum apolipoprotein C-III (apoCIII) concentration and apoCIII gene polymorphisms have been shown to be a risk factor for cardiovascular disease; however, the underlying mechanisms remain unclear. In addition, no studies have been performed that address these issues in type 1 diabetes. The current study investigated apoCIII protein and apoCIII gene variation in a normotriglyceridemic (82 +/- 57 mg/dL) population of patients with type 1 diabetes, the Diabetes Control and Complications Trial/Epidemiology of Diabetes Intervention and Complications (DCCT/EDIC) cohort. Blood samples were obtained in 409 patients after an overnight fast. Serum apoCIII concentration was highly correlated with multiple changes in lipids and lipoproteins that resulted in an adverse cardiovascular disease risk profile. Higher apoCIII concentrations were associated (P <.0001) with increased triglycerides (r = 0.78), total (r = 0.61) and low-density lipoprotein (LDL) (r = 0.40) cholesterol, apoA-I (r = 0.26), and apoB (r = 0.50), and these relationships persisted after controlling for age, gender, body mass index (BMI), and hemoglobin A1c (HbA1c). Nuclear magnetic resonance (NMR) lipoprotein subclass analyses demonstrated that apoCIII was correlated with an increase in very-low-density lipoprotein (VLDL) subclasses (P = .0001). There also was a highly significant positive relationship between serum apoCIII concentration and the LDL particle concentration in both men (r = 0.49, P = .001) and women (r = 0.40, P = .001), and a highly significant negative relationship between serum apoCIII levels and average LDL particle size in both men (r = -0.37, P = .001) and women (r = -0.22, P = .001) due primarily to an augmentation in the small L1 subclass (r = 0.42, P = .0001). Neither the T(-455) --> C polymorphism affecting an insulin response element in the apoCIII gene promoter nor a SacI polymorphism in the 3'UTR were associated with any alterations in circulating apoCIII concentrations, serum lipids, apolipoprotein concentrations, lipoprotein composition, or parameters measured by NMR lipoprotein subclass analyses. In summary, elevated apoCIII concentration was associated with risk factors for cardiovascular disease in normolipidemic type 1 diabetic patients through associated changes in lipoprotein subfraction distributions, which were independent of apoCIII genotype.

Greater enrichment of triacylglycerol-rich lipoproteins with apolipoproteins E and C-III after meals rich in saturated fatty acids than after meals rich in unsaturated fatty acids

Background: Although there is considerable interest in the postprandial events involved in the absorption of dietary fats and the subsequent metabolism of diet-derived triacylglycerol-rich lipoproteins, little is known about the effects of meal fatty acids on the composition of these particles. Objective: We examined the effect of meal fatty acids on the lipid and apolipoprotein contents of triacylglycerol-rich lipoproteins. Design: Ten normolipidemic men received in random order a mixed meal containing 50 L, of a mixture of palm oil and cocoa butter [rich in saturated fatty acids (SFAs)], safflower oil [n-6 polyunsaturated fatty acids (PUFAs)]. or olive oil [monounsaturated fatty acids (MUFAs)] on 3 occasions. Fasting and postprandial apolipoproteins B-48. B-100, E. C-II, and C-III and lipids (triacylglycerol and cholesterol) were measured in plasma fractions with Svedberg flotation rates (S-f) >400 S-f 60-400, and S-f 20 - 60. Results: Calculation of the composition of the triacylglycerol-rich lipoproteins (expressed per mole of apolipoprotein B) showed notable differences in the lipid and apolipoprotein contents of the SFA-enriched particles in the S-f > 400 and S-f 60-400 fractions. After the SFA meal, triacylglycerol-rich lipoproteins in these fractions showed significantly greater amounts of triacylglycerol and of apolipoproteins C-II (Sf 60-400 fraction only), C-III, and E than were found after the MUFA meal (P < 0.02) and more cholesterol, apolipoprotein C-III (Sf > 400 fraction only), and apolipoprotein E than after the PUFA meal (P < 0.02). Conclusions: Differences in the composition of S-f > 400 and S-f 60-400 triacylglycerol-rich lipoproteins formed after saturated compared with unsaturated fatty acid-rich meals may explain differences in the metabolic handling of dietary fats.

Energy levels, radiative rates and electron impact excitation rates for transitions in C III

We report energy levels, radiative rates (A-values) and lifetimes for the astrophysically important Be-like ion C III. For the calculations, 166 levels belonging to the n ≤ 5 configurations are considered and the GRASP (General-purpose Relativistic Atomic Structure Package) is adopted. Einstein A-coefficients are provided for all E1, E2, M1 and M2 transitions, while lifetimes are compared with available measurements as well as theoretical results, and no large discrepancies noted. Our energy levels are assessed to be accurate to better than 1 per cent for a majority of levels, and A-values to better than 20 per cent for most transitions. Collision strengths are also calculated, for which the Dirac Atomic R-matrix Code (DARC) is used. A wide energy range, up to 21 Ryd, is considered and resonances resolved in a fine energy mesh in the thresholds region. The collision strengths are subsequently averaged over a Maxwellian velocity distribution to determine effective collision strengths up to a temperature of 8.0 ×10[5]K, sufficient for most astrophysical applications. Our data are compared with the recent R-matrix calculations of Fernández-Menchero et al., and significant differences (up to over an order of magnitude) are noted for several transitions over the complete temperature range of the results.

The works of Martin Doyle [pseud.] containing I. Hints to small farmers on land, fences, cottages, potatoes, mangel wurzel, turnips, manures, &c. II. Hints on road-work, ventilation, health, dress, temperance, education, &c. III. Hints on planting, cattle, fowls, fisheries, agricultural implements, flax, &c. IV. Irish cottagers.

Each work has also special t-p. and titles vary from those on main t.-p. I. Hints originally intended for small farmers of the county of Wexford ... 7th ed., rev. and enl. 1831. II. Hints addressed to the small holders and peasantry of Ireland on road-making and on ventilation, &c. &c. 1830. III. Hints to small holders on planting and on cattle. &c &c. 1830. IV. Irish cottagers ... 1830.

A compendious system of anatomy : In six parts. Part I. Osteology. II. Of the muscles, &c. III. Of the abdomen. Part IV. Of the thorax. V. Of the brain and nerves. VI. Of the senses. From the Encyclopaedia. Illustrated with twelve large copperplates.

Austin

Iconografía de la moneda griega de Sicilia (siglos VI a. C. - III a. C.)

Esta tesis versa sobre el estudio y análisis de los tipos iconográficos que aparecen en las monedas sicilianas acuñadas durante la etapa de presencia griega en la isla (siglos VI a.C. – III a.C.) hasta la inclusión total de la isla bajo dominio de Roma. Este estudio y análisis de sus diferentes tipos iconográficos abarca todas sus cecas, así como también estudia las acuñaciones de la población de Reghio, por tratarse de una zona de influencia directa de la moneda griega de Sicilia. Esta tesis no se trata de un catálogo nuevo o revisión de los antiguos, sino que trata sobre la relación histórica así como la influencia cultural y religiosa que pueden ser observadas a través de las imágenes acuñadas en las monedas griegas. Por lo tanto, además de realizar un estudio iconográfico, trato de encontrar la relación por la cual ese motivo en concreto es elegido así como sus posibles influencias según el área geográfica en el que se acuña o según la población a la que representa (principalmente griega, élima, sícula o cartaginesa). Además, utilizo los datos aportados por las excavaciones de los yacimientos arqueológicos para poder observar y determinar las posibles relaciones entre los cultos allí realizados y la moneda local, tratando de hacer hincapié en la presencia de santuarios y cultos adscritos según las fuentes antiguas y la arqueología. Hay que añadir que, además de tratar los yacimientos como fuente documental, también analizo la presencia de ocultamientos o tesoros monetarios encontrados en Sicilia para hacer una relación de éstos con su contexto histórico, así como también analizo la metrología de las monedas sicilianas a lo largo de este periodo histórico, para comprobar si corresponde el área de influencia ponderal con el área de influencia iconográfico. La estructura de la tesis se compone de dos partes fundamentalmente, un catálogo revisado de todas las cecas sicilianas durante los siglos VI a.C. – III a.C. y un análisis iconográfico de los elementos que aparecen representados en ellas...

Experimental and computational studies on lipoprotein lipolysis at acidic pH

Atherosclerosis is a disease of the arteries; its characteristic features include chronic inflammation, extra- and intracellular lipid accumulation, extracellular matrix remodeling, and an increase in extracellular matrix volume. The underlying mechanisms in the pathogenesis of advanced atherosclerotic plaques, that involve local acidity of the extracellular fluid, are still incompletely understood. In this thesis project, my co-workers and I studied the different mechanisms by which local extracellular acidity could promote accumulation of the atherogenic apolipoprotein B-100 (apoB-100)-containing plasma lipoprotein particles in the inner layer of the arterial wall, the intima. We found that lipolysis of atherogenic apoB-100-containing plasma lipoprotein particles (LDL, IDL, and sVLDL) by the secretory phospholipase A2 group V (sPLA2-V) enzyme, was increased at acidic pH. Also, the binding of apoB-100-containing plasma lipoprotein particles to human aortic proteoglycans was dramatically enhanced at acidic pH. Additionally, lipolysis by sPLA2-V enzyme further increased this binding. Using proteoglycan-affinity chromatography, we found that sVLDL lipoprotein particles consist of populations, differing in their affinities toward proteoglycans. These populations also contained different amounts of apolipoprotein E (apoE) and apolipoprotein C-III (apoC-III); the amounts of apoC-III and apoE per particle were highest in the population with the lowest affinity toward proteoglycans. Since PLA2-modification of LDL particles has been shown to change their aggregation behavior, we also studied the effect of acidic pH on the monolayer structure covering lipoprotein particles after PLA2-induced hydrolysis. Using molecular dynamics simulations, we found that, in acidity, the monolayer is more tightly packed laterally; moreover, its spontaneous curvature is negative, suggesting that acidity may promote lipoprotein particles fusion. In addition to extracellular lipid accumulation, the apoB-100-containing plasma lipoprotein particles can be taken up by inflammatory cells, namely macrophages. Using radiolabeled lipoprotein particles and cell cultures, we showed that sPLA2-V-modification of LDL, IDL, and sVLDL lipoproteins particles, at neutral or acidic pH, increased their uptake by human monocyte-derived macrophages.

Hepatic denervation impairs the assembly and secretion of VLDL-TAG

VLDL secretion is a regulated process that depends on the availability of lipids, apoB and MTP. Our aim was to investigate the effect of liver denervation upon the secretion of VLDL and the expression of proteins involved in this process. Denervation was achieved by applying a 85% phenol solution onto the portal tract, while control animals were treated with 9% NaCl. VLDL secretion was evaluated by the Tyloxapol method. The hepatic concentration of TAG and cholesterol, and the plasma concentration of TAG, cholesterol, VLDL-TAG, VLDL-cholesterol and HDL-cholesterol were measured, as well as mRNA expression of proteins involved in the process of VLDL assembly. Hepatic acinar distribution of MTP and apoB was evaluated by immunohistochemistry. Denervation increased plasma concentration of cholesterol (125.3 +/- 10.1 vs. 67.1 +/- 4.9 mg dL(-1)) and VLDL-cholesterol (61.6 +/- 5.6 vs. 29.4 +/- 3.3 mg dL(-1)), but HDL-cholesterol was unchanged (45.5 +/- 6.1 vs. 36.9 +/- 3.9 mg dL(-1)). Secretion of VLDL-TAG (47.5 +/- 23.8 vs. 148.5 +/- 27.4 mg dL h(-1)) and mRNA expression of CPT I and apoB were reduced (p < 0.01) in the denervated animals. MTP and apoB acinar distribution was not altered in the denervated animals, but the intensity of the reaction was reduced in relation to controls. Copyright (C) 2008 John Wiley & Sons, Ltd.

A proteome signature for intrauterine growth restriction derived from multifactorial analysis of mass spectrometry-based cord blood serum profiling

Intrauterine growth restriction (IUGR) is defined as a condition in which the fetus does not reach its genetically given growth potential, resulting in low birth weight. IUGR is an important cause of perinatal morbidity and mortality, thus contributing substantially to medically indicated preterm birth in order to prevent fetal death. We subjected umbilical cord blood serum samples either belonging to the IUGR group (n = 15) or to the control group (n = 15) to fractionation by affinity chromatography using a bead system with hydrophobic interaction capabilities. So prepared protein mixtures were analyzed by MALDI-TOF mass spectrometric profiling. The six best differentiating ion signals at m/z 8205, m/z 8766, m/z 13 945, m/z 15 129, m/z 15 308, and m/z 16 001 were collectively assigned as IUGR proteome signature. Separation confidence of our IUGR proteome signature reached a sensitivity of 0.87 and a specificity of 0.93. Assignment of ion signals in the mass spectra to specific proteins was substantiated by SDS-PAGE in conjunction with peptide mass fingerprint analysis of cord blood serum proteins. One constituent of this proteome signature, apolipoprotein C-III(0) , a derivative lacking glycosylation, has been found more abundant in the IUGR cord blood serum samples, irrespective of gestational age. Hence, we suggest apolipoprotein C-III(0) as potential key-marker of the here proposed IUGR proteome signature, as it is a very low-density lipoprotein (VLDL) and high-density lipoprotein (HDL) member and as such involved in triglyceride metabolism that itself is discussed as being of importance in IUGR pathogenesis. Our results indicate that subtle alterations in protein glycosylation need to be considered for improving our understanding of the pathomechanisms in IUGR.

Low concentrations of diacylglycerol promote the binding of apolipophorin III to a phospholipid bilayer: a surface plasmon resonance spectroscopy study.

The binding of the exchangeable apolipoprotein apolipophorin III (apoLp-III) to an egg phosphatidylcholine bilayer as a function of the concentration of diacylglycerol (DG) in the bilayer was studied by surface plasmon resonance spectroscopy. At a DG concentration of 2 mol % in the bilayer, the binding of apoLp-III reached saturation. Under saturating conditions, apoLp-III forms a closely packed monolayer approximately 55 A thick, in which each molecule of protein occupies approximately 500 A2 at the membrane surface. These dimensions are consistent with the molecular size of the apoLp-III molecule determined by x-ray crystallography, if apoLp-III binds to the bilayer with the long axis of the apoLp-III normal to the membrane surface. In the absence of protein, the overall structure of the lipid bilayer was not significantly changed up to 2.5 mol% DG. However, at 4 and 6 mol % DG, the presence of nonbilayer structures was observed. The addition of apoLp-III to a membrane containing 6 mol % DG promoted the formation of large lipid-protein complexes. These data support a two-step sequential binding mechanism for binding of apoLp-III to a lipid surface. The first step is a recognition process, consisting of the adsorption of apoLp-III to a nascent hydrophobic defect in the phospholipid bilayer caused by the presence of DG. This recognition process might depend on the presence of a hydrophobic sensor located at one of the ends of the long axis of the apoLp-III molecule but would be consolidated through H-bond and electrostatic interactions. Once primary binding is achieved, subsequent enlargement of the hydrophobic defect in the lipid surface would trigger the unfolding of the apolipoprotein and binding via the amphipathic alpha-helices. This two-step sequential binding mechanism could be a general mechanism for all exchangeable apolipoproteins. A possible physiological role of the ability of apoLp-III to bind to lipid structures in two orientations is also proposed.

Basic equations for suspension flows with interphase mass-transfer

In the case of suspension flows, the rate of interphase momentum transfer M(k) and that of interphase energy transfer E(k), which were expressed as a sum of infinite discontinuities by Ishii, have been reduced to the sum of several terms which have concise physical significance. M(k) is composed of the following terms: (i) the momentum carried by the interphase mass transfer; (ii) the interphase drag force due to the relative motion between phases; (iii) the interphase force produced by the concentration gradient of the dispersed phase in a pressure field. And E(k) is composed of the following four terms, that is, the energy carried by the interphase mass transfer, the work produced by the interphase forces of the second and third parts above, and the heat transfer between phases. It is concluded from the results that (i) the term, (-alpha-k-nabla-p), which is related to the pressure gradient in the momentum equation, can be derived from the basic conservation laws without introducing the "shared-pressure presumption"; (ii) the mean velocity of the action point of the interphase drag is the mean velocity of the interface displacement, upsilonBAR-i. It is approximately equal to the mean velocity of the dispersed phase, upsilonBAR-d. Hence the work terms produced by the drag forces are f(dc) . upsilonBAR-d, and f(cd) . upsilonBAR-d, respectively, with upsilonBAR-i not being replaced by the mean velocity of the continuous phase, upsilonBAR-c; (iii) by analogy, the terms of the momentum transfer due to phase change are upsilonBAR-d-GAMMA-c, and upsilonBAR-d-GAMMA-d, respectively; (iv) since the transformation between explicit heat and latent heat occurs in the process of phase change, the algebraic sum of the heat transfer between phases is not equal to zero. Q(ic) and Q(id) are composed of the explicit heat and latent heat, so that the sum Q(ic) + Q(id)) is equal to zero.

Proteómica dirigida en la cirugía del cáncer gástrico

240 p. + anexos

在辉光放电中八氟环丁烷聚合的研究

I 理论部分：对未知结构聚合物的ESCA谱进行表征，一是利用已知结构小分子化合物的ESCA谱中结合能值进行标定，二是运用电荷电位模型经量子化学计算对谱峰进行归属。由于所处化学环境不同，用小分子化合物的ESCA谱峰位对象合物标定不够准确，量子化学计算又太复杂。本工作提出了电负性经验公式，可计算ESCA谱中C_(1S)结合能位移值，与实测值作对照即可判断聚合物结构，方法简便，准确可靠。电负性经验公式表达如下：聚合物分子链上某碳原子1S电子结合能位移ΔEc可看作该碳原子与相邻基团间电负性差值和的函数，即ΔEc = K_c ∑ from i=1 to 4 1/(n_i) (X_i - 2.5) + b。聚合物分子链上各碳原子的实际电负性为X_c = (∑ form j=1 to 4 1/(n_j) X_j/∑ form j=1 to 4 1/(n_j) + 2.5)/2运用电负性经验公式计算了九个模型化合物中C_(1S)的ΔEc，推导出F作为α、β、γ位碳上的取代基的对α位碳上1s电子结合能位移影响产生的效应值，即，对于ΔEc = aα + bβ + cγ，有α = 3.018 (±0.007)，β = 0.2513 (±0.0007) γ = 0.0176 (±0.003)。此结果与ESCA谱实测值相符。本工作设计了十六种化合物模型，模型中具有含氟等离子体聚合物中可能存在的大部分基团，通过电负性经验公式计算求得化学环境不同的C_(1S)结合能位移值，把ESCA谱的实测值和计算值进行对照，可对象合物的求知结构作出大致判断。II 在辉光放电中八氟环丁烷的聚合与刻蚀反应：在碳氧化合物的辉光放电中同时存在聚合与刻蚀两种相反的过程，聚合与刻蚀竞争的结果对聚合速率，产物结构都有很大影响。本工作通过测定聚合速率、IR谱、ESCA谱研究了反应条件对八氟环丁烷等离子体聚合与刻蚀的影响，得到了一致的和半定量的结果。实验证实，随着功率的增大或压力的减小，刻蚀有增强的趋势，不同的等离子气体对聚合与刻蚀的竞争过程有不同的影响，刻蚀强弱的顺序为Ar>N_2>H_2>He，增加等离子气体对单体的比率，引起刻蚀增加。聚合速率不随功率的改变而变化的状态是由聚合与刻蚀间的平衡引起的。随着功率增大或压力减少，聚合产物支化程度增加，线性程度减小，F/C比相应减小。同样随等离子气体种类不同，聚合物支化程度大小的顺序是Ar>N_2>H_2>He，F/C比相应变化。总之，聚合产物支化程度的变化和聚合与刻蚀的竞争结果相一致。对等离子体聚合与刻蚀竞争过程的定量描述未见文献报导。III 八氟环丁烷等离子体聚合物的结构和表征：用IR和ESR谱测得OFCB等离子体聚合物中存在五种元素：C、F、O、N、Si，并对可能存在于结构中的基因作了分析。通过NaHSO_3的亲核双键加成反应，用IR和ESR谱证实了等离子体聚合物中存在的双键及其类型。类似工作未见文献报导。用CuSO_4·5H_2作标样，测出PPOFCB中的自由基浓度为10~(19)自旋数/克。在80、100、120、140 ℃考察PPOFCB中俘获自由基的衰减过程，证实此过程涉及两类不同的反应，一是自由基的复合反应，二是自由基的氧气的终止反应。对上述两类反应进行动力学处理，求出了自由基复合反应在不同温度时的速度常数，并得到反应治化能为374焦耳/摩尔或8.869千卡/摩尔。氧和自由基的终止反应中，氧气的扩散过程是决定反应速度的步骤，反应速度常数就是扩散速度常数，K = 5.3 * 10~(-23)自旋数~(-1)、克分~(-1)。根据VPO测出的PPOFCB的分子量和自由基浓度，算出每4.5个分子中有一个自由基。类似工作未见文献报导。PPOFCB的分子量随反应条件不同而变化，用VPO测出，在80W和40W功率，在Ar存在下制备的OFCB等离子体聚合物的分子量分别为2736和2003。用~(19)F-NMR谱对PPOFCB中的基团进行定量计算，得出在一个90W，He气中制备的聚合物分子中，CF_3-CF_2为6.1，CF_2为13.5，CF_3-CF为7.9，CF为17.5，CF_3-C为6.1，C为13.5。在用上述方法进行结构表征的基础上，提出了OFCB等离子体聚合物的结构模型。IV 八氟环丁烷等离子体聚合的反应历程：以Ar、N_2、H_2、He为等离子气体，分别对OFCB等离子体聚合的气体冷凝物进行GC-Mg、~(19)F-NMR和ESR测试，观察到在气相中，OFCB经历了等离子态反应。单体被打断的程度，和高能电子浓度有关。体系中电子浓度依赖于气体的第一电离势和气相内的化学反应类型。初级粒子二氟卡宾：CF_2和中间体六氟丙烯是气相中的主要产物。~(19)F-NMR证实气相中存在支化产物，表明聚合物在气相中发生了重排。ESR谱测得，在气相产物中存在高浓度的由二种以上组分混合而成的自由基，自由基间的复合反应是生成聚合物的主要增长过程，气相产物的分子量随放电功率的升高而增大。根据聚合物中存在的高含量CF_3基团，分析了CF_3基团对聚合过程的影响。CF_3基团产生了屏蔽效应，它既保护了聚合物不致被降解，也限制了生成高分子量产物。实验证实，CF_3基团的生长来自F的重排，同时未自支化气相产物的非均相增长。OFCB等离子体聚合物中俘获自由基主要是由高能电子轰击聚合物表面产生的，同时来自被固相吸附的带有自由基的气相产物。在碳氟化合物的等离子体聚合中，吸附是和聚合同样重要的过程，气相产物被吸附后，可继续再引发、增长、刻蚀等反应过程、直到成为固相内层分子为止。ESR谱证实，高能粒子的轰击在辉光放电中起主导作用，紫外辐射对含氟聚合物引发和脱氟作用微弱。在气相和表面反应历程讨论的基础上，提出了OFCB等离子体聚合的反应历程模型，解释了碳氢和碳氟两个等离子体象合体系遵循不同反应历程的原因。本部分工作的实验和反应历程解释基本未见文献报导。