We have inactivated the endogenous apolipoprotein E (apoE) gene by using targeting in mouse embryonic stem (ES) cells. Two plasmids were used, pJPB63 and pNMC109, both containing a neomycin-resistance that replaces part of apoE disrupts its structure. ES cell colonies targeted after electroporation with plasmid identified polymerase chain reaction (PCR) followed genomic Southern analysis. Of 648 G418-resistant analyzed, 9 gave positive signal PCR amplification, 5 them confirmed as blot The...

The vascular endothelium mediates the ability of blood vessels to alter their architecture in response hemodynamic changes; however, specific endothelial-derived factors that are responsible for remodeling poorly understood. Here we show nitric oxide (NO) is a major mediator controlling remodeling. In external carotid artery ligation, mice with targeted disruption endothelial synthase gene (eNOS) did not remodel ipsilateral common arteries whereas wild-type did. Rather, eNOS mutant displayed...

Apolipoprotein E-deficient mice have spontaneous elevations of total plasma cholesterol and triglycerides reduced high-density lipoprotein. The develop arterial lesions in a time-dependent manner. Lesional distribution was centered the aortic sinus young mice, were widely distributed throughout tree at 8 to 9 months age. In subendothelial foam cell deposits present adjacent valve-attachment sites. By 5 age, deposits, free cholesterol, admixed smooth muscle cells composed developing...

Variants of the human angiotensinogen gene have been linked in some studies to increased circulating levels and essential hypertension. To test for direct causality between genotypes at locus blood pressures, we studied mice carrying zero, one, two, three, or four functional copies murine wild-type (Agt) its normal chromosomal location. Plasma increase progressively, although not linearly, from zero zero-copy animals 145% four-copy animals. Mice all are birth, but most die before weaning....

The renin-angiotensin system plays a critical role in sodium and fluid homeostasis. Genetic or acquired alterations the expression of components this are strongly implicated pathogenesis hypertension. To specifically examine physiological genetic functions type 1A receptor for angiotensin II, we have disrupted mouse gene encoding embryonic stem cells by targeting. Agtr1A(-/-) mice were born expected numbers, histomorphology their kidneys, heart, vasculature was normal. AT1 receptor-specific...

Studies by various investigators have indicated that elevated levels of plasma homocyst(e)ine are strongly associated with the occurrence occlusive vascular diseases. With eventual aim determining whether or not concentrations directly causative cardiovascular diseases, we generated mice moderately and severely homocyst(e)inemic. Homologous recombination in mouse embryonic stem cells was used to inactivate cystathionine beta-synthase [L-serine hydrolyase (adding homocysteine), EC 4.2.1.22]...

In recent years there has been an accumulation of evidence for the existence nanobubbles on hydrophobic surfaces in water, despite predictions that such small bubbles should rapidly dissolve because high internal pressure associated with interfacial curvature and resulting increase gas solubility. Nanobubbles are interest among surface scientists their potential importance long-range attraction, microfluidics, adsorption at surfaces. Here we employ recently developed techniques designed to...

Apolipoprotein E (apo E)-deficient mice are severely hypercholesterolemic and develop advanced atheromas independent of diet. The C57BL/6 strain differs from most inbred strains by having lower HDL concentrations a high risk developing early atherosclerotic lesions when fed an atherogenic relative deficiency atherosclerosis susceptibility the corrected with expression human apolipoprotein AI AI) transgene in this genetic background. To examine if increases apo also effective minimizing...

Interfacial nanobubbles (INBs) on a solid surface in contact with water have drawn widespread research interest. Although several theoretical models been proposed to explain their apparent long lifetimes, the underlying mechanism still remains dispute. In this work, morphological evolution of INBs was examined air-equilibrated and partially degassed use atomic force microscopy (AFM). Our results show that (1) shrank while they grew slightly water, (2) three-phase boundary pinned during INBs....

The adhesion and friction of smooth polymer surfaces were studied below the glass transition temperature by use a surface forces apparatus. force crosslinked was orders magnitude less than that an uncrosslinked polymer. In contrast, after chain scission outermost layers, hysteresis increase substantially. These results show polymer-polymer depend on dynamic rearrangement segments at shearing interfaces, both as is made from to with long chains quasi-free ends. suggest new ways for...

We describe a general way of introducing transgenes into the mouse germ line for comparing different sequences without complications variation in copy number and insertion site. The method uses homologous recombination embryonic stem (ES) cells to generate mice having single transgene integrated chosen location genome. To test method, murine bcl-2 cDNA driven by either chicken beta-actin promoter or human has been inserted immediately 5' X-linked hypoxanthine phosphoribosyltransferase locus...

We have used a "plug and socket" targeting technique to generate mouse model of beta 0-thalassemia in which both the b1 b2 adult globin genes been deleted. Mice homozygous for this deletion (Hbbth-3/Hbbth-3) die perinatally, similar most severe form Cooley anemia humans. heterozygous appear normal, but their hematologic indices show characteristics typical thalassemia, including dramatically decreased hematocrit, hemoglobin, red blood cell counts, mean corpuscular volume, hemoglobin...

With the aim of establishing whether a genetically reduced capability producing apolipoprotein E (apo E) can affect atherogenesis, we have compared consequences dietary stress on normal mice and heterozygous or homozygous for disrupted apo gene. A dramatically accelerated development lesions occurred in vasculature mutants as result feeding an atherogenic diet 12 wk, extensive deposition lipid-filled macrophages was found outside cardiovascular system. In nine heterozygotes fed amount their...

Embryonic stem cells (derived from 129/Ola mice) containing a mutant hypoxanthine phosphoribosyltransferase gene that had been corrected in vitro planned manner by homologous recombination were injected into blastocysts obtained C57BL/6J mice. The introduced pseudopregnant female mice to complete their development. Eleven surviving pups obtained. Nine chimeras: six males and three females. Two of the transmitted embryonic cell genome alteration offspring at high frequencies. These...

The hypoxanthine-guanine phosphoribosyltransferase (Hprt) gene has been mutated in mouse blastocyst-derived embryonic stem cells by site-directed homologous recombination. Embryonic were electroporated the presence of a targeting DNA fragment containing two specific features: (i) contained promoterless neomycin phosphotransferase (neo) that, when located within endogenous Hprt locus, could be transcribed from promoter target locus. (ii) had short regions homology with gene: one, 132 base...

The consequences of the lack apolipoprotein A-I (apoA-I) were evaluated in mice made to apoA-I by gene targeting. Inbred strain 129 homozygous for inactive Apoa1 and maintained on regular mouse chow had markedly reduced total cholesterol (26% normal) high-density lipoprotein (HDL) (25% levels their plasma. Their plasma lipoproteins lacked all other apolipoproteins but apoE. ApoE comprises about one third protein HDL particles homozygotes, whereas it is present only trace amounts normal HDL....

Previous studies have identified lipid-poor high density lipoproteins with electrophoretic pre-beta mobility as the initial acceptors of cell-derived cholesterol in human plasma. These contain apolipoprotein A-I (apo A-I) their sole apolipoprotein. In present study, incubation plasma [3H]cholesterol-laden skin fibroblasts has led to identification another lipoprotein that serves a potent acceptor cholesterol. This lipoprotein, which we term gamma-LpE, exhibits gamma on agarose gel...

Thirty-three kilobase pairs (kb) of human DNA containing the haptoglobin (Hp) and haptoglobin-related (Hpr) gene pair were cloned, nucleotide sequence 21-kb was determined. The two genes are closely linked, with Hpr being 2.2 kb downstream Hp. Six hundred nucleotides occur between that not found either 5' to Hp or 3' gene. After duplication event, first intron acquired a 9-kb insert consisting mainly retrovirus-like element potential primer-binding site homologous mouse isoleucine tRNA....

The two major components of greenhouse gases, CO2 and water, are indispensable for sustaining life on Earth. Water vapor is the most significant gas that has provided earth with an "atmospheric blanket" prevented surface from freezing. However, contemporary climate models largely consider influence water as a factor within positive feedback loops, while possibility direct anthropogenic emissions primary drivers global warming remains underexplored. In particular, common assumption been...

Apolipoprotein E (apoE)-deficient mice develop marked hyperlipidemia as well atherosclerosis and thus are an excellent animal model for evaluating the potential gene therapy in human genetic dyslipoproteinemias. Recombinant adenovirus containing either apoE (rAdv.apoE) or reporter luciferase (rAdv.luc) were generated infused intravenously apoE-deficient with preinfusion plasma total cholesterol of 644 +/- 149 mg/dl rich VLDL/IDL. After a single infusion rAdv.apoE, concentrations ranging from...

Generation of oxidative products by phagocytic cells is known to be an important host defense mechanism directed toward killing invading microorganisms. The importance two major oxidant-producing enzymes, myeloperoxidase (MPO) and NADPH-oxidase, in vivo fungicidal action was directly compared genetically engineered mice. Both MPO-deficient (MPO−/−) NADPH-oxidase-deficient (X-linked chronic granulomatous disease [X-CGD]) mice showed increased susceptibility pulmonary infections with Candida...

The formation and disappearance of liquid bridges between two surfaces can occur either through equilibrium or nonequilibrium processes. In the first instance, bridge molecules are in thermodynamic with surrounding vapor medium. second, chemical potential gradients result material transfer; mechanical instabilities, because van der Waals force jumps on approach a Rayleigh instability rapid separation, may trigger irreversible film coalescence snapping. We have studied growth mechanisms...

The cDNA for the 1,25-dihydroxyvitamin D3 receptor has been isolated by screening a rat kidney lambda gt11 expression library with mixture of three previously identified anti-receptor monoclonal antibodies. Protein produced from this reacts each antibodies but does not react an antibody specific porcine or irrelevant antibody. In addition, expressed protein specifically binds bind estradiol, progesterone, testosterone, cortisol, aldosterone. nucleotide sequence determined and used to deduce...

Dietary administration of probucol (0.5%, wt/wt) efficiently reduced total plasma cholesterol levels in apolipoprotein E-deficient mice (apoE-/-) by 40%, with decreases high density lipoprotein (HDL) and apoAI 70 50%, respectively. Paradoxically, however, aortic atherosclerotic plaques the probucol-treated apoE-/- formed more rapidly than untreated mice, lesions were two to four times larger mature regardless sex, age, genetic background (P < 10(-)6). Histologically, contained increased...