Actinide Crystallization and Fission Reactions in Cooling White Dwarf Stars.

The first solids that form as a cooling white dwarf (WD) starts to crystallize are expected to be greatly enriched in actinides. This is because the melting points of WD matter scale as Z^{5/3} and actinides have the largest charge Z. We estimate that the solids may be so enriched in actinides that they could support a fission chain reaction. This reaction could ignite carbon burning and lead to the explosion of an isolated WD in a thermonuclear supernova (SN Ia). Our mechanism could potentially explain SN Ia with sub-Chandrasekhar ejecta masses and short delay times.

Elasticity of Nuclear Pasta.

The elastic properties of neutron star crusts are relevant for a variety of currently observable or near-future electromagnetic and gravitational wave phenomena. These phenomena may depend on the elastic properties of nuclear pasta found in the inner crust. We present large-scale classical molecular dynamics simulations where we deform nuclear pasta. We simulate idealized samples of nuclear pasta and describe their breaking mechanism. We also deform nuclear pasta that is arranged into many domains, similar to what is known for the ions in neutron star crusts. Our results show that nuclear pasta may be the strongest known material, perhaps with a shear modulus of 10^{30} ergs/cm^{3} and a breaking strain greater than 0.1.

Disordered nuclear pasta, magnetic field decay, and crust cooling in neutron stars.

Nuclear pasta, with nonspherical shapes, is expected near the base of the crust in neutron stars. Large-scale molecular dynamics simulations of pasta show long lived topological defects that could increase electron scattering and reduce both the thermal and electrical conductivities. We model a possible low-conductivity pasta layer by increasing an impurity parameter Q_{imp}. Predictions of light curves for the low-mass x-ray binary MXB 1659-29, assuming a large Q_{imp}, find continued late time cooling that is consistent with Chandra observations. The electrical and thermal conductivities are likely related. Therefore, observations of late time crust cooling can provide insight on the electrical conductivity and the possible decay of neutron star magnetic fields (assuming these are supported by currents in the crust).

Gastric H+/K(+)-ATPase: targeting signals in the regulation of physiologic function.

The H+/K(+)-ATPase of gastric parietal cells catalyzes acid secretion in the stomach. Regulation of this pump's function involves its targeting to an intracellular storage compartment, which fuses with the apical plasma membrane in response to secretagogue stimulation. The H+/K(+)-ATPase must also be returned to this intracellular storage compartment to inactivate acid secretion. Recent research suggests that a number of targeting signals and protein-protein interactions participate in governing this process.

Structure of multicomponent Coulomb crystals.

Coulomb plasmas crystallize in a number of physical systems, such as dusty plasmas, neutron star crusts, and white dwarf cores. The crystal structure of the one-component and binary plasma has received significant attention in the literature, though the less studied multicomponent plasma may be most relevant for many physical systems which contain a large range of particle charges. We report on molecular dynamics simulations of multicomponent plasmas near the melting temperature with mixtures taken to be realistic x-ray burst ash compositions. We quantify the structure of the crystal with the bond order parameters and radial distribution function. Consistent with past work, low charge particles form interstitial defects and we argue that they are in a quasiliquid state within the lattice. The lattice shows screening effects which preserves long-range order despite the large variance in particle charges, which may impact transport properties relevant to astrophysics.

An ion-transporting ATPase encodes multiple apical localization signals.

Epithelial cells accumulate distinct populations of membrane proteins at their two plasmalemmal domains. We have examined the molecular signals which specify the differential subcellular distributions of two closely related ion pumps. The Na,K-ATPase is normally restricted to the basolateral membranes of numerous epithelial cell types, whereas the H,K-ATPase is a component of the apical surfaces of the parietal cells of the gastric epithelium. We have expressed full length and chimeric H,K-ATPase/Na,K-ATPase cDNAs in polarized renal proximal tubular epithelial cells (LLC-PK1). We find that both the alpha and beta subunits of the H,K-ATPase encode independent signals that specify apical localization. Furthermore, the H,K-ATPase beta-subunit possesses a sequence which mediates its participation in the endocytic pathway. The interrelationship between epithelial sorting and endocytosis signals suggested by these studies supports the redefinition of apical and basolateral as functional, rather than simply topographic domains.

A transmembrane segment determines the steady-state localization of an ion-transporting adenosine triphosphatase.

The H,K-adenosine triphosphatase (ATPase) of gastric parietal cells is targeted to a regulated membrane compartment that fuses with the apical plasma membrane in response to secretagogue stimulation. Previous work has demonstrated that the alpha subunit of the H, K-ATPase encodes localization information responsible for this pump's apical distribution, whereas the beta subunit carries the signal responsible for the cessation of acid secretion through the retrieval of the pump from the surface to the regulated intracellular compartment. By analyzing the sorting behaviors of a number of chimeric pumps composed of complementary portions of the H, K-ATPase alpha subunit and the highly homologous Na,K-ATPase alpha subunit, we have identified a portion of the gastric H,K-ATPase, which is sufficient to redirect the normally basolateral Na,K-ATPase to the apical surface in transfected epithelial cells. This motif resides within the fourth of the H,K-ATPase alpha subunit's ten predicted transmembrane domains. Although interactions with glycosphingolipid-rich membrane domains have been proposed to play an important role in the targeting of several apical membrane proteins, the apically located chimeras are not found in detergent-insoluble complexes, which are typically enriched in glycosphingolipids. Furthermore, a chimera incorporating the Na, K-ATPase alpha subunit fourth transmembrane domain is apically targeted when both of its flanking sequences derive from H,K-ATPase sequence. These results provide the identification of a defined apical localization signal in a polytopic membrane transport protein, and suggest that this signal functions through conformational interactions between the fourth transmembrane spanning segment and its surrounding sequence domains.

Localization of Na+,K+-ATPase alpha-subunit to the sinusoidal and lateral but not canalicular membranes of rat hepatocytes.

Controversy has recently developed over the surface distribution of Na+,K+-ATPase in hepatic parenchymal cells. We have reexamined this issue using several independent techniques. A monoclonal antibody specific for the endodomain of alpha-subunit was used to examine Na+,K+-ATPase distribution at the light and electron microscope levels. When cryostat sections of rat liver were incubated with the monoclonal antibody, followed by either rhodamine or horseradish peroxidase-conjugated goat anti-mouse secondary, fluorescent staining or horseradish peroxidase reaction product was observed at the basolateral surfaces of hepatocytes from the space of Disse to the tight junctions bordering bile canaliculi. No labeling of the canalicular plasma membrane was detected. In contrast, when hepatocytes were dissociated by collagenase digestion, Na+,K+-ATPase alpha-subunit was localized to the entire plasma membrane. Na+,K+-ATPase was quantitated in isolated rat liver plasma membrane fractions by Western blots using a polyclonal antibody against Na+,K+-ATPase alpha-subunit. Plasma membranes from the basolateral domain of hepatocytes possessed essentially all of the cell's estimated Na+,K+-ATPase catalytic activity and contained a 96-kD alpha-subunit band. Canalicular plasma membrane fractions, defined by their enrichment in alkaline phosphatase, 5' nucleotidase, gamma-glutamyl transferase, and leucine aminopeptidase had no detectable Na+,K+-ATPase activity and no alpha-subunit band could be detected in Western blots of these fractions. We conclude that Na+,K+-ATPase is limited to the sinusoidal and lateral domains of hepatocyte plasma membrane in intact liver. This basolateral distribution is consistent with its topology in other ion-transporting epithelia.

Susceptibility to antibiotics of Bacillus anthracis strains isolated in Romania.

A number of 21 B. anthracis strains isolated from 16 pustules, 2 blood cultures and 3 cerebrospinal fluids during 2000-2004 were studied for their susceptibility to antibiotics. The antibiosusceptibility testing was performed by disk diffusion method, on Mueller-Hinton agar medium. Two of the studied strains exhibited resistance to penicillins, considered until not long ago as antimicrobial agents of choice for the treatment of anthrax infection. The penicillin resistance explained the difficulties encountered during the treatment of these two cases as well as the fatal evolution in one of them. Both penicillin-resistant strains were subsequently tested, by using "in agar" antibiotic dilution method, in order to determine the minimum inhibitory concentrations (MIC) of the respective strains to penicillin G by the help of a serial antibiotic dilution from 16 microg/ml to 0.0075 microg/ml. The MIC values were 0.5 microg/ml and 4 microg/ml respectively, whereas in case of the standard B. anthracis 34F2 Sterne strain was < 0.015 microg/ml. All the 21 B. anthracis tested strains exhibited resistance to the IIIrd generation cephalosporins, as well as to TMP/STX, but were sensitive to tetracyclines and fluoroquinolones, these sensitivity aspects coming into agreement with the literature data. The strains proved to be also susceptible as follows: 13 strains to macrolides, 15 to rifampicin, 16 to chloramphenicol and all 21 to gentamycin; the last antibiotic can be used in association with fluoroquinolones in the treatment of B. anthracis infections. Fluoroquinolones (i.e. ciprofloxacin) become drugs of choice for the treatment of B. anthracis infections if early administered (within the first 24 hrs), in advance of the germ invasion into the lymph system and septicemia, preventing in this way the bacterial multiplication and production of edemathogenic and lethal toxins.

Necrotizing enterocolitis comes in different forms: Historical perspectives and defining the disease.

The specific cause of what is commonly referred to as necrotizing enterocolitis (NEC) disease has been elusive largely because it is becoming clear that this entity represents more than one disease with multifactorial pathogenic mechanisms. Furthermore, finding clear and consistent diagnostic biomarkers will be difficult until the different subsets of what we are calling this disease are better delineated. In this introductory chapter, we discuss different disease entities that are frequently termed "NEC" in the newborn infant. We hope this will set the stage for more focused research and development of preventative measures for at least the most common forms of this disease.

The NH(2)-terminus of norepinephrine transporter contains a basolateral localization signal for epithelial cells.

When expressed in epithelial cells, dopamine transporter (DAT) was detected predominantly in the apical plasma membrane, whereas norepinephrine transporter (NET) was found in the basolateral membrane, despite 67% overall amino acid sequence identity. To identify possible localization signals responsible for this difference, DAT-NET chimeras were expressed in MDCK cells and localized by immunocytochemistry and transport assays. The results suggested that localization of these transporters in MDCK cells depends on their highly divergent NH(2)-terminal regions. Deletion of the first 58 amino acids of DAT (preceding TM1) did not change its apical localization. However, the replacement of that region with corresponding sequence from NET resulted in localization of the chimeric protein to the basolateral membrane, suggesting that the NH(2)-terminus of NET, which contains two dileucine motifs, contains a basolateral localization signal. Mutation of these leucines to alanines in the context of a basolaterally localized NET/DAT chimera restored transporter localization to the apical membrane, indicating that the dileucine motifs are critical to the basolateral localization signal embodied within the NET NH(2)-terminal region. However, the same mutation in the context of wild-type NET did not disrupt basolateral localization, indicating the presence of additional signals in NET directing its basolateral localization within the plasma membrane.

Hypoxia increases stimulus-induced PAF production and release from human umbilical vein endothelial cells.

Hypoxia alters endothelial cell function and metabolism. Since PAF is synthesized by endothelial cells and capable of modulating endothelial cell responses, we investigated the effect of hypoxia on synthesis and release of PAF from endothelial cells. We found: (1) Approx. 90% of the radylPAF derivative in stimulated endothelial cells is acylPAF. (2) Acute hypoxic (15 min-1 h) priming increased ionophore- and thrombin-induced radylPAF accumulation. (3) Long-term hypoxic exposure increased radylPAF accumulation at 24 and 48 h in the presence of ionophore. (4) Bioactive PAF was released into media and hypoxia and ionophore synergistically increased PAF release. (5) Hypoxia and ionophore stimulation increased phospholipase A2 activity and decreased acetylhydrolase activity in endothelial cells. We conclude that hypoxia and ionophore increase PAF synthesis and release from endothelial cells.

DNMT-dependent suppression of microRNA regulates the induction of GBM tumor-propagating phenotype by Oct4 and Sox2.

Cancer stem-like cells represent poorly differentiated multipotent tumor-propagating cells that contribute disproportionately to therapeutic resistance and tumor recurrence. Transcriptional mechanisms that control the phenotypic conversion of tumor cells lacking tumor-propagating potential to tumor-propagating stem-like cells remain obscure. Here we show that the reprogramming transcription factors Oct4 and Sox2 induce glioblastoma cells to become stem-like and tumor-propagating via a mechanism involving direct DNA methyl transferase (DNMT) promoter transactivation, resulting in global DNA methylation- and DNMT-dependent downregulation of multiple microRNAs (miRNAs). We show that one such downregulated miRNA, miRNA-148a, inhibits glioblastoma cell stem-like properties and tumor-propagating potential. This study identifies a novel and targetable molecular circuit by which glioma cell stemness and tumor-propagating capacity are regulated.

The cell biology of ion pumps: sorting and regulation.

The physiologic function of an ion pump is determined, in part, by its subcellular localization and by the cellular mechanisms that modulate its activity. The Na,K-ATPase and the gastric H,K-ATPase are two closely related members of the P-type family of ion transporting ATPases. Despite their homology, these pumps are sorted to different domains in polarized epithelial cells and their enzymatic activities are subject to distinct regulatory pathways. The molecular signals responsible for these properties have begun to be elucidated. It appears that a complex array of inter- and intra-molecular interactions govern these proteins' trafficking, distribution and catalytic capacity.

Fulminant multiple system atrophy in a young adult presenting as motor neuron disease.

A 34-year-old man demonstrated rapidly progressive motor neuron disease and, late in his 9-month clinical course, exhibited ophthalmoplegia and dysautonomic symptoms. Neuropathology showed spinal and bulbar motor neuron disease with severe involvement of extraocular motor nuclei, degeneration of spinal sympathetic and bulbar parasympathetic nuclei, striatonigral degeneration, and early olivopontocerebellar atrophy. This case underscores the diversity of multiple system atrophy and demonstrates an unusually rapid course in a young patient.

Polarized distribution of Na+,K+-ATPase in giant cells elicited in vivo and in vitro.

Giant cell formation was analyzed to determine whether it results in the high level of Na+,K+-ATPase expression that characterizes multinucleated cells such as osteoclasts. Giant cells and fusing alveolar macrophages were subjected to morphological, immunological, and biochemical studies. Both subunits of the Na+,K+-ATPase were found to be present on the plasma membrane of giant cells. Their localization was restricted to the non-adherent domain of the cell surface. Dynamic studies of giant cell differentiation demonstrated that on culture and/or multinucleation, an increase in sodium pump alpha-subunit synthesis occurred and led to a high level of expression of Na pumps. Conversely, the adherent plasma membrane of giant cells was enriched in a lysosomal membrane antigen. This study demonstrates that culture and/or multinucleation induces a significant increase in the expression of sodium pumps. The polarized distribution of these pumps and of a lysosomal component suggests that fusing macrophages undergo biochemical and morphological alterations which prepare them for a new and specialized function in chronic inflammatory reactions. Giant cells may offer a suitable model system to study the differentiation of other related multinucleated cells, such as osteoclasts.

Immunocytochemical localization of plasmalemmal proteins in semi-thin sections of epithelial monolayers.

We describe a technique for analysis by light microscopic immunocytochemistry of the distribution of plasmalemmal proteins in polarized epithelial cells. For this purpose, Madin Darby Canine Kidney (MDCK) cells were grown to confluency on Cytodex beads, the beads were fixed with formaldehyde, and semi-thin (0.5 micron) sections were cut at liquid nitrogen temperature on an ultracryomicrotome. The distribution of the basolaterally distributed plasmalemmal protein, Na,K-ATPase, was assessed by indirect immunofluorescence using a monospecific polyclonal antibody directed against the alpha-subunit of the Na pump. Such preparations enable epithelial monolayers to be evaluated in cross-section, thus permitting unambiguous topological assessment of apical and basolateral membrane proteins. Thus, the spatial uncertainties encountered in en face examination of membrane protein distribution in epithelia grown on solid supports are largely obviated. In addition, we describe a technique for removal of the bead matrix, which markedly reduces nonspecific background staining and improves access of reagents to the basal cell surface, thus permitting localization of basal lamina components.

The dimensions of the triangle of Koch in children.

We measured the dimensions of Koch's triangle in children with normal intracardiac anatomy to determine the relation between the size of the triangle of Koch and patient age, weight, height, and body surface area. We found that the dimensions of Koch's triangle varies significantly and directly with patient age and body habitus in this pediatric population.

Regulation of myocardial glucose uptake and transport during ischemia and energetic stress.

Myocardial glucose utilization increases in response to the energetic stress imposed on the heart by exercise, pressure overload, and myocardial ischemia. Recruitment of glucose transport proteins is the cellular mechanism by which the heart increases glucose transport for subsequent metabolism. Moderate regional ischemia leads to the translocation of both glucose transporters, GLUT4 and GLUT1, to the sarcolemma in vivo. Myocardial ischemia also stimulates 5'-adenosine monophosphate-activated protein kinase, which may be a fuel gauge in the heart and other tissues signaling the need to turn on energy-generating metabolic pathways. Pharmacologic stimulation of this kinase increases cardiac glucose uptake and transporter translocation, suggesting that it may play an important role in augmenting glucose entry in the setting of ischemic or energetic stress. Thus, recent work has provided insight into the cellular and molecular mechanisms responsible for glucose uptake during energetic stress, which may lead to new approaches to the treatment of patients with coronary artery disease.

Platelet-activating factor increases mucosal permeability in rat intestine via tyrosine phosphorylation of E-cadherin.

1. Platelet-activating factor (PAF), an inflammatory mediator, plays an important role in mediating intestinal injury. However, it remains unclear whether PAF has a function in the intestine. The production of PAF by normal intestine and by unstimulated intestinal epithelial cell lines suggests that PAF may have a regulatory function in the normal bowel. 2. In this study we investigated the role of PAF in modulating intestinal mucosal permeability in rats. Lumen-to-blood transit of FD-4 (dextran 4400), (an index of intestinal permeability), was assessed in sham-operated rats and rats injected with PAF (1.25 microg kg(-1), i.v., a dose insufficient to induce intestinal injury). 3. PAF-induced villus cytoskeletal changes were examined by staining the intestine for F-actin. The effect of PAF on tyrosine phosphorylation of the junctional protein E-cadherin was examined by immunoprecipitation. Some rats were pretreated with AG1288 (a tyrosine kinase inhibitor) before PAF injection, and mucosal permeability change was assessed. 4. To investigate the role of endogenous PAF upon mucosal permeability, we studied the effect of PAF antagonists on (intraluminal) glucose-induced increase in mucosal permeability. 5. We found that low dose PAF: (a) alters the cytoskeletal structure of intestinal epithelium, (b) causes the influx of FD4 from intestinal lumen to systemic circulation, (c) induces tyrosine phosphorylation of E-cadherin and cadherin-associated proteins. Glucose-induced mucosal permeability increase is abolished by using two structurally different PAF antagonists. 6. These results suggest that endogenous PAF modulates macromolecular movement across the intestinal mucosal barrier, probably via tyrosine phosphorylation of E-cadherin and cytoskeletal alteration of enterocytes.