Variation in lipoprotein(a) concentrations among individuals with the same apolipoprotein (a) isoform is determined by the rate of lipoprotein(a) production

Lipoprotein(a) [Lp(a)] is an atherogenic lipoprotein which is similar in structure to, but metabolically distinct from, LDL. Factors regulating plasma concentrations of Lp(a) are poorly understood. Apo(a), the protein that distinguishes Lp(a) from LDL, is highly polymorphic, and apo(a) size is inversely correlated with plasma Lp(a) level. Even within the same apo(a) isoform class, however, plasma Lp(a) concentrations vary widely. A series of in vivo kinetic studies were performed using purified radiolabeled Lp(a) in individuals with the same apo(a) isoform but different Lp(a) levels. In a group of seven subjects with a single S4-apo(a) isoform and Lp(a) levels ranging from 1 to 13.2 mg/dl, the fractional catabolic rate (FCR) of 131I-labeled S2-Lp(a) (mean 0.328 day-1) was not correlated with the plasma Lp(a) level (r = -0.346, P = 0.45). In two S4-apo(a) subjects with a 10-fold difference in Lp(a) level, the FCR's of 125I-labeled S4-Lp(a) were very similar in both subjects and not substantially different from the FCRs of 131I-S2-Lp(a) in the same subjects. In four subjects with a single S2-apo(a) isoform and Lp(a) levels ranging from 9.4 to 91 mg/dl, Lp(a) concentration was highly correlated with Lp(a) production rate (r = 0.993, P = 0.007), but poorly correlated with Lp(a) FCR (mean 0.304 day-1). Analysis of Lp(a) kinetic parameters in all 11 subjects revealed no significant correlation of Lp(a) level with Lp(a) FCR (r = -0.53, P = 0.09) and a strong correlation with Lp(a) production rate (r = 0.99, P < 0.0001). We conclude that the substantial variation in Lp(a) levels among individuals with the same apo(a) phenotype is caused primarily by differences in Lp(a) production rate.     

A Psi of relief.

The American Psychological Association (APA) introduced a new logo symbol for the association in 1991. The symbol attempted to modernize the representation of the organization, melding the clinical and experimental arms of the discipline through the use of organic arms attached to a central unifying pole. To determine what this symbol represented to a naive audience, the authors undertook a questionnaire-based investigation with a total of 176 undergraduates. The most common images evoked were a sunrise, a disguised version of the AT&T logo, a window, an organizational logo (not APA), a plant, a human face, a bird, or a tool or implement. The authors discuss these interpretations as symbols for the future goals of APA. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Apolipoprotein (a)

Apo (a) consists of multiple tandem repeat of kringle 4, which resembles a counterpart of plasminogen. Plasma Lp (a) levels are genetically determined primarily by alleles at the apo (a) gene. Apo (a) shows size heterogeneity on the analysis of the protein and the mRNA. Pulse field gel electrophoresis revealed that size heterogeneity is largely due to different numbers of kringle 4-encoding sequences in the apo (a) gene. The cloning of apo (a) gene and identification of the 5'-flanking promoter region provide tools to study the regulation of apo (a) gene. The development of transgenic mice expressing human apo (a) offered a good model for understanding of atherosclerosis associated with elevated plasma levels of Lp (a).     

Interactions between the structural domains of the RNA replication proteins of plant-infecting RNA viruses

Brome mosaic virus (BMV), a positive-strand RNA virus, encodes two replication proteins: the 2a protein, which contains polymerase-like sequences, and the 1a protein, with N-terminal putative capping and C-terminal helicase-like sequences. These two proteins are part of a multisubunit complex which is necessary for viral RNA replication. We have previously shown that the yeast two-hybrid assay consistently duplicated results obtained from in vivo RNA replication assays and biochemical assays of protein-protein interaction, thus permitting the identification of additional interacting domains. We now map an interaction found to take place between two 1a proteins. Using previously characterized 1a mutants, a perfect correlation was found between the in vivo phenotypes of these mutants and their abilities to interact with wild-type 1a (wt1a) and each other. Western blot analysis revealed that the stabilities of many of the noninteracting mutant proteins were similar to that of wt1a. Deletion analysis of 1a revealed that the N-terminal 515 residues of the 1a protein are required and sufficient for 1a-1a interaction. This intermolecular interaction between the putative capping domain and itself was detected in another tripartite RNA virus, cucumber mosaic virus (CMV), suggesting that the 1a-1a interaction is a feature necessary for the replication of tripartite RNA viruses. The boundaries for various activities are placed in the context of the predicted secondary structures of several 1a-like proteins of members of the alphavirus-like superfamily. Additionally, we found a novel interaction between the putative capping and helicase-like portions of the BMV and CMV 1a proteins. Our cumulative data suggest a working model for the assembly of the BMV RNA replicase.     

Environment-dependent tight-binding potential model

The yellow-pigmented bacterium isolated from a ditch was a gram negative rod with a G+C content of 63 mol%, and was classified in the genus Sphingomonas. Electron microscopy revealed that the bacterial cell surface was covered with many large plaits. When grown in a medium containing a polysaccharide as an essential nutrient, a pit of 0.02-0.1 micrometers in diameter was formed on the cell surface, and a thin section showed the rearrangement of the plaits and the presence of a region where the cell membrane sinks into the cytosol. The dependence of the pit formation on the presence of macromolecule may predict the existence of a direct uptake mechanism for macromolecules through a mouth-like pit, possibly in endocytosis fashion. The confirmation of the pit structure is the first such finding in the history of microbiology and may provide a new insight into the cell morphology and biochemistry of macromolecule transport in microbial cell system.     

Effects of TCDD on Ah receptor, ARNT, EGF, and TGF-alpha expression in embryonic mouse urinary tract

Lipoprotein(a) [Lp(a)] biogenesis was examined in primary cultures of hepatocytes isolated from mice transgenic for both human apolipoprotein(a) [apo(a)] and human apoB. Steady-state and pulse-chase labeling experiments demonstrated that newly synthesized human apo(a) had a prolonged residence time (approximately 60 min) in the endoplasmic reticulum (ER) before maturation and secretion. Apo(a) was inefficiently secreted by the hepatocytes and a large portion of the protein was retained and degraded intracellularly. Apo(a) exhibited a prolonged and complex folding pathway in the ER, which included incorporation of apo(a) into high molecular weight, disulfide-linked aggregates. These folding characteristics could account for long ER residence time and inefficient secretion of apo(a). Mature apo(a) bound via its kringle domains to the hepatocyte cell surface before appearing in the culture medium. Apo(a) could be released from the cell surface by apoB-containing lipoproteins. These studies are consistent with a model in which the efficiency of post-translational processing of apo(a) strongly influences human plasma Lp(a) levels, and suggest that cell surface assembly may be one pathway of human Lp(a) production in vivo. Transgenic mouse hepatocytes thus provide a valuable model system with which to study factors regulating human Lp(a) biogenesis.     

Imprinting, learning, and memory.

If a visually naive chick is exposed to one of a wide range of conspicuous objects, the chick may learn its characteristics. A series of biochemical studies has implicated a restricted part of the forebrain in this process of imprinting; a specific region (IMHV) has been identified which may be a site of information storage. Changes in the morphology of synapses occur in this region as a consequence of training. The left and right IMHV regions play different roles in the imprinting process. Exposure to a simple artificial object, a rotating red box, has different neural consequences from those associated with exposure to a complex object, a rotating stuffed jungle fowl, which resembles a conspecific. These differences may be related to the differences in complexity of the two training objects. Another possibility is that two neural systems are implicated in imprinting: (a) a system that underlies a predisposition to approach objects resembling conspecifics and (b) a learning system, of which IMHV is a crucial component, that is engaged by particular objects and that in "natural" circumstances also allows the chick to learn the characteristics of its mother. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Amyloid beta-peptide spin trapping. II: Evidence for decomposition of the PBN spin adduct

We investigated the effect of lipoprotein(a) (Lp(a)) on proliferation of human arterial smooth muscle cells (SMCs) and its mechanisms of action. Low density lipoprotein (LDL), Lp(a) and apolipoprotein(a) (apo(a)) significantly stimulated the proliferation of SMCs. Lp(a) and apo(a) reduced the amount of active transforming growth factor-beta (TGF-beta) with the mink lung epithelial cell bioassay, however LDL had no effect. Lp(a), but not apo(a), significantly stimulated the proliferation of SMCs even in the presence of a neutralizing antibody for TGF-beta. Our results suggest that Lp(a) stimulates the proliferation of SMCs via apo(a)-induced inhibition of TGF-beta activation and stimulation of SMCs by the LDL-particle of Lp(a).     

Concept-Based Method for Extracting Valid Subsets from an EXPRESS Schema

An EXPRESS schema is a data schema defined in EXPRESS, an international standard language for defining product data schemas. This technical paper proposes and formally defines a set of conditions for generating a minimum valid subset of an EXPRESS schema corresponding to a concept, where a concept is a general idea and a subset is a partial model of a data schema. We introduce a notion of “minimal set” to define the relationships between a subset and other subsets, and also between a subset and concepts. A minimal set is the smallest complete subset of a schema that corresponds to a concept. Using IFC, an international standard data model for the architecture, engineering, and construction industry, the proposed conditions have been implemented in a software application developed for extracting subsets from the IFC schema matching the concepts. A number of examples are demonstrated.     

Dynamics of mechanical action on materials. X. Hydraulic loading of a system

The movement of an open mechanical system with one degree of freedom modelling uniaxial tension of a viscoelastic strain hardenable body in a machine with a hydraulic drive that generates a variable force and is simultaneously a damping element of the system is analyzed. It is demonstrated that transient movement of the system in question corresponds to a fourth order non-autonomous dynamic system that has stable periodic and aperiodic solutions in relation to its control parameters. In view of a marked decrease in viscous resistance of the system caused by the presence of a hydraulic drive aperiodic damping is typical for the transient process. It is shown that a system with a hydraulic element differs markedly from a system that does not contain such an element.     

Dynamic interaction between a coherent precipitate and an edge dislocation

Dynamic interaction between a coherent precipitate and an edge dislocation is analyzed by means of a discrete atom method, which is based on classical statistical mechanics and linear elasticity. Precipitates having a dilatational misfit strain and elastic constants different from those of the matrix phase are treated in anisotropic elastic systems under a plane strain condition. A coherent interface transforms into a semicoherent one by nucleating dipolar dislocations at a stress concentration in a coherent precipitate. One of the dipolar dislocations glides along the precipitate-matrix interface to become a misfit dislocation, and the other slips into the matrix phase to become a lattice dislocation. In accordance with continuum elasticity, a coherent particle with a positive misfit strain migrates to the tension side of an edge dislocation, whereas a particle with a negative misfit diffuses to the region of compression. Morphological change is, however, caused by the dislocation as the particle tries to capitalize on the dislocation stress field, and the particle shape depends on its stiffness and elastic anisotropy. Under an applied shear, a hard coherent particle with a positive misfit strain is sheared along the shear direction, but a soft particle responds in the opposite direction. Elastic interaction between a coherent particle and an edge dislocation can be so strong that the particle-dislocation complex remains coupled even at a high shear strain applied to the system. Some composite applied stresses can cause an edge dislocation to split into two partials. One of the partials, a glissile component, is found to engage actively in the morphological evolution of a particle during a diffusional relaxation. This article is based on a presentation made in the symposium “Kinetically Determined Particle Shapes and the Dynamics of Solid:Solid Interfaces,” presented at the October 1996 Fall meeting of TMS/ASM in Cincinnati, Ohio, under the auspices of the ASM Phase Transformations Committee.     

Oxidation of apolipoprotein(a) inhibits kringle-associated lysine binding: the loss of intrinsic protein fluorescence suggests a role for tryptophan residues in the lysine binding site

Lipoprotein(a) [Lp(a)] is a low-density lipoprotein complex consisting of apolipoprotein(a) [apo(a)] disulfide-linked to apolipoprotein B-100. Lp(a) has been implicated in atherogenesis and thrombosis through the lysine binding site (LBS) affinity of its kringle domains. We have examined the oxidative effect of 2,2'-azobis-(amidinopropane) HCl (AAPH), a mild hydrophilic free radical initiator, upon the ability of Lp(a) and recombinant apo(a), r-apo(a), to bind through their LBS domains. AAPH treatment caused a time-dependent decrease in the number of functional Lp(a) or r-apo(a) molecules capable of binding to fibrin or lysine-Sepharose and in the intrinsic protein fluorescence of both Lp(a) and r-apo(a). The presence of a lysine analogue during the reaction prevented the loss of lysine binding and provided a partial protection from the loss of tryptophan fluorescence. The partial protection of fluorescence by lysine analogues was observed in other kringle-containing proteins, but not in proteins lacking kringles. No significant aggregation, fragmentation, or change in conformation of Lp(a) or r-apo(a) was observed as assessed by native or SDS-PAGE, light scattering, retention of antigenicity, and protein fluorescence emission spectra. Our results suggest that AAPH destroys amino acids in the kringles of apo(a) that are essential for lysine binding, including one or more tryptophan residues. The present study, therefore, raises the possibility that the biological roles of Lp(a) may be mediated by its state of oxidation, especially in light of our previous study showing that the reductive properties of sulfhydryl-containing compounds increase the LBS affinity of Lp(a) for fibrin.     

C3a is a chemotaxin for human eosinophils but not for neutrophils. I. C3a stimulation of neutrophils is secondary to eosinophil activation

Inflammatory action of the potent chemotaxin C5a has been well characterized on a variety of human cell types, including neutrophils, monocytes, basophils, and eosinophils. The cellular effects of C3a are less well defined. Contradictory reports have been published for C3a activation of neutrophils. Recent reports that C3a activates both basophils and eosinophils prompted us to reinvestigate the effects of C3a stimulation on eosinophils. We hypothesized that C3a activation of eosinophils, cells that are present in most neutrophil preparations, might lead to neutrophil activation. Using neutrophils of 98% purity, we observed no evidence of cellular activation after stimulation with either C3a, recombinant human C3a (rhC3a), or the synthetic C3a analogue C3a 57-77, Y57. Eosinophils purified to > 98% purity displayed concentration-dependent polarization, chemotaxis, and enzyme release by stimulation with C3a, rhC3a, and the synthetic C3a analogue. An inactive form of C3a, C3adesArg, failed to stimulate either eosinophils or neutrophils. Using neutrophil preparations containing 5-9% eosinophils, up to 20% of neutrophils became polarized after exposure to C3a. Likewise, we demonstrated that supernatant from C3a-stimulated eosinophils promotes neutrophil chemotaxis. Eosinophil polarization experiments were repeated in the presence of antibody to the C5a receptor (C5aR) to show that C3a and C5a interact with different receptors. C3a activates eosinophils in the presence of anti-C5aR antibody at concentrations that fully block C5a activation. We conclude that eosinophils are directly activated by either C3a or C5a, whereas C3a failed to activate neutrophils. C3a acts on eosinophils via a receptor that is distinct from C5aR. Since neutrophils are indirectly stimulated by C3a, eosinophils contaminating neutrophil preparations may explain earlier reports that C3a activates human neutrophils.     

Postprocessing the Hybrid Method for Addressing Uncertainty in Risk Assessments

In a previous paper in this Journal, a “hybrid method” was proposed for the joint propagation of probability distributions (expressing variability) and possibility distributions (i.e., fuzzy numbers, expressing imprecision or partial ignorance) in the computation of risk. In order to compare the results of the hybrid computation (a random fuzzy set) to a tolerance threshold (a tolerable level of risk), a postprocessing method was proposed. Recent work has highlighted a shortcoming of this postprocessing step which yields overly conservative results. A postprocessing method based on Shafer’s theory of evidence provides a rigorous answer to the problem of comparing a random fuzzy set with a threshold. The principles behind the new postprocessing scheme are presented and illustrated with a synthetic example.     

Inhibition of N-linked glycosylation results in retention of intracellular apo[a] in hepatoma cells, although nonglycosylated and immature forms of apolipoprotein[a] are competent to associate with apolipoprotein B-100 in vitro

Apolipoprotein[a] (apo[a]) is a highly polymorphic glycoprotein that forms a covalent complex with apolipoprotein B-100 (apoB-100), producing a lipoprotein species referred to as lipoprotein[a] (Lp[a]). We have studied the effects of alterations in glycosylation of apo[a] on its intracellular processing and secretion as well as its ability to associate with low density lipoprotein (LDL) apoB-100. HepG2 cells transfected with a 6 kringle IV (6 K-IV) apo[a] minigene were treated with tunicamycin, an inhibitor of N-linked glycosylation, which eliminated apo[a]-B-100 complexes from the media. Tunicamycin treatment also reduced secretion of the 6 K-IV apo[a] protein from transfected McA-RH7777 cells by approximately 50%, but completely eliminated secretion of apo[a] species containing 9 and 17 K-IV repeats. Mixing experiments, performed with radiolabeled media (+/-tunicamycin) from transfected McA-RH7777 cells, demonstrated no alteration in the extent of association of apo[a] with human LDL. Similar mixing experiments using culture media from glycosylation-defective mutant chinese hamster ovary (CHO) cells transfected with the same apo[a] minigene showed identical results. Apo[a] secretion was demonstrated in all mutant cell lines in the absence of either N- or O-linked (or both) glycosylation. The mechanisms underlying the reduced secretion of apo[a] from transfected hepatoma cells were examined by pulse-chase radiolabeling and apo[a] immunoprecipitation. Tunicamycin treatment altered the efficiency of precursor apo[a] processing from the ER by increasing its ER retention time. The increased accumulation of precursor apo[a] in the ER was associated with alterations in the kinetics of association with two resident endoplasmic reticulum (ER) chaperone proteins, calnexin and BiP. These findings suggest that the glycosylation state and size of apo[a] appear to play a role in regulating its efficient exit from the endoplasmic reticulum. However, neither N- nor O-linked glycosylation of apo[a] exerts a major regulatory role in its covalent association with apoB-100.     

Utility and feasibility of assessing myocardial mechanics after bypass

The authors present the case of a 34-year-old man struck over the left mastoid region by a hockey puck, who suffered a fatal rupture of a left vertebral artery berry aneurysm. He became apneic within seconds of the injury and had no brainstem reflex within minutes. The postmortem examination showed massive subarachnoid hemorrhage in the posterior fossa and the remnants of a berry aneurysm near the intradural origin of the left vertebral artery, 11 mm proximal to the posterior inferior cerebellar artery. Rupture of a saccular aneurysm as a result of head trauma is rare. This is the first reported case of a posterior circulation aneurysm rupture as a result of head trauma.     

Transition to Self-Similarity of Diffusion of Tracer in Turbulent Patch

A mixing of a passive tracer inside a turbulent patch generated by a localized short-time perturbation is studied numerically and analytically. Two kinds of an initial distribution of a tracer are considered: two-layer and continuous with constant gradient. For the turbulent patch shaped as a layer, it is shown that, regardless of details of initial distributions of a turbulent energy and dissipation, a tracer concentration evolves to self-similar regimes as time elapses. Analytical self-similar solutions to turbulent diffusion equations are found for three symmetric shapes of a turbulent patch: layer, cylinder, and sphere. Distributions of the concentration inside a patch are found to be substantially nonuniform, with a typical ratio of a concentration gradient in the middle of a patch to its initial value of about 0.5.     

Neomycin inhibits secretion of apolipoprotein[a] by increasing retention on the hepatocyte cell surface

Neomycin therapy reduces plasma levels of low density lipoprotein and lipoprotein[a] (Lp[a]). To determine whether neomycin directly alters the biogenesis of Lp[a], we have examined the effect of neomycin on apolipoprotein[a] (apo[a]) synthesis and secretion in primary cultures of baboon hepatocytes. Using this system, we have previously shown that apo[a] is synthesized as a lower molecular weight precursor that upon maturation becomes associated with the cell surface before release into the culture medium. Treatment of hepatocytes with 10 mM neomycin reduced levels of apo[a] in the culture medium by as much as 12-fold. Although a portion of the reduced secretion could be accounted for by a reduction in total protein synthesis, the greatest effect of neomycin on apo[a] secretion was to decrease the release of mature apo[a] from the hepatocyte cell surface into the culture medium. Treatment of hepatocyte cultures with trypsin confirmed that mature apo[a] in neomycin-treated cells was still transported to the cell surface. Examination of related antibiotics demonstrated that inhibition of apo[a] secretion is a general property shared by the deoxystreptamine antibiotics. The mechanism by which neomycin affects the apo[a]-cell surface interaction is not known, but neomycin is known to perturb cell surface membranes, inhibit the interaction of some ligands with their cell surface receptors, and inhibit the metabolism of phosphatidylinositol 4,5 biphosphate. These studies suggest that cell surface association of apo[a] may play a role in Lp[a] biogenesis in vivo.