bluuebunny/crossref_metadata_2025_split huggingface dataset:
In vivo expression of human ATP:cob(I)alamin adenosyltransferase (ATR) using recombinant adeno‐associated virus (rAAV) serotypes 2 and 8
Authors: Erger Kirsten E. , Conlon Thomas J. , Leal Nicole A. , Zori Robert , Bobik Thomas A. , Flotte Terence R. 
Published: May 2007

AbstractBackgroundMethylmalonic aciduria (MMA) is an autosomal recessive disease with symptoms that include ketoacidosis, lethargy, recurrent vomiting, dehydration, respiratory distress, muscular hypotonia and death due to methylmalonic acid levels that are up to 1000‐fold greater than normal. CblB MMA, a subset of the mutations leading to MMA, is caused by a deficiency in the enzyme cob(I)alamin adenosyltransferase (ATR). No animal model currently exists for this disease. ATR functions within the mitochondria matrix in the final conversion of cobalamin into coenzyme B12, adenosylcobalamin (AdoCbl). AdoCbl is a required coenzyme for the mitochondrial enzyme methylmalonyl‐CoA mutase (MCM).MethodsThe human ATR cDNA was cloned into a recombinant adeno‐associated virus (rAAV) vector and packaged into AAV 2 or 8 capsids and delivered by portal vein injection to C57/Bl6 mice at a dose of 1 × 1010 and 1 × 1011 particles. Eight weeks post‐injection RNA, genomic DNA and protein were then extracted and analyzed.ResultsUsing primer pairs specific to the cytomegalovirus (CMV) enhancer/chicken β‐actin (CBAT) promoter within the rAAV vectors, genome copy numbers were found to be 0.03, 2.03 and 0.10 per cell in liver for the rAAV8 low dose, rAAV8 high dose and rAAV2 high dose, respectively. Western blotting performed on mitochondrial protein extracts demonstrated protein levels were comparable to control levels in the rAAV8 low dose and rAAV2 high dose animals and 3‐ to 5‐fold higher than control levels were observed in high dose animals. Immunostaining demonstrated enhanced transduction efficiency of hepatocytes to over 40% in the rAAV8 high dose animals, compared to 9% and 5% transduction in rAAV2 high dose and rAAV8 low dose animals, respectively.ConclusionsThese data demonstrate the feasibility of efficient ATR gene transfer to the liver as a prelude to future gene therapy experiments. Copyright © 2007 John Wiley & Sons, Ltd.
Switching on transgene expression by correcting aberrant splicing using multi‐targeting steric‐blocking oligonucleotides
Authors: Resina Sarah , Kole Ryszard , Travo Adrian , Lebleu Bernard , Thierry Alain R. 
Published: May 2007

AbstractBackgroundMutations leading to aberrant splicing are found as a cause of numerous pathologies. Splice‐switching oligonucleotides (SSOs), which modify aberrant expression patterns of alternatively spliced mRNAs, are a novel means of potentially controlling such diseases.MethodsWe used an experimental model in which a mutated β‐globin intron, carrying an aberrant splice site at nucleotide 705, interrupts the coding region of the luciferase reporter gene inserted in HeLa pLuc/705 cells. We have optimized delivery of splice correcting, steric‐blocking 2′‐O‐methyl SSOs targeting the 705 mutated region (2′‐O‐Me SSO705) with DLS (DLS: delivery liposomal system) lipoplexes.ResultsOptimal luciferase activity for DLS/2′‐O‐Me SSO705 was achieved at 100 nM and was detectable at concentrations as low as 10 nM in serum‐containing culture medium, confirming the potential of DLS lipoplex‐mediated nuclear SSO delivery as observed in cellular uptake studies. We confirmed by cytofluorometry and epifluorescence microscopy the high potential of the DLS lipoplex for cellular and nuclear oligonucleotide uptake. The DLS lipoplex was then used to directly compare the intracellular efficacy of various SSO chemistries and sequences in correction of aberrant splicing. 2′‐O‐Methoxyethyl‐oligodeoxyribonucleoside phosphorothioates had a greater activity than 2′‐O‐methyl phosphodiester or 2′‐O‐methyl‐phosphorothioate oligoribonucleotides. Targeting the splicing enhancer 623 region upstream was as efficient as targeting the 705 splice site, and, remarkably, simultaneous targeting of both sites was more efficient than treatment of the cells either with 2′‐O‐Me SSO705 or 2′‐O‐Me SSO623 alone.ConclusionsWe demonstrated that SSOs can switch on luciferase activity in HeLa cells previously transfected with the pLuc/705 plasmid via the same DLS vector and provides a novel approach to modulate the expression of a transgene. Copyright © 2007 John Wiley & Sons, Ltd.
Human osteocalcin: a strong promoter for nitric oxide synthase gene therapy, with specificity for hormone refractory prostate cancer
Authors: McCarthy Helen O. , Coulter Jonathan A. , Worthington Jenny , Robson Tracy , Hirst David G. 
Published: May 2007

AbstractBackgroundGene therapy has been identified as a promising treatment strategy for hormone refractory prostate cancer (HRPC). We report, for the first time, the use of the human osteocalcin (hOC) promoter to control inducible nitric oxide synthase (iNOS) transgene expression in HRPC.MethodsHuman prostate carcinoma cells (PC3, DU145, LNCaP), colon cancer cells (HT29) and human microvascular endothelial cells (HMEC‐1) were transfected in vitro with constitutively driven CMV/iNOS or hOC/iNOS plasmid DNA by cationic lipid vector. End points of these experiments were Western blotting, NO· generation using the Greiss test to measure accumulated nitrite, and clonogenic assay.ResultsTransfection of the hOC/iNOS plasmid increased iNOS protein and total nitrite levels in PC3 and DU145 cells, but not LNCaP or HT29. Transfection with CMV/iNOS or hOC/iNOS resulted in no additional cytotoxicity in androgen‐dependent LNCaP cells or in the non‐prostate cell lines. However, transfection with either construct resulted in a greatly reduced cell survival (to 10–20%) in the androgen‐independent PC3 and DU145 cell lines.ConclusionsUtilising the tumour‐type specific properties of the hOC promoter in tandem with the iNOS gene, we have demonstrated target cell specificity, and transgene activation, in the androgen‐independent prostate cancer cell lines (PC3 and DU145), an effect absent in normal and androgen‐dependent cells. Furthermore, the levels of NO· generated are comparable with those seen generated with constitutively (CMV)‐driven iNOS. The data obtained from this study provide a basis for future development of hOC/iNOS gene therapy. Copyright © 2007 John Wiley & Sons, Ltd.
Comparison of cDNA and genomic forms of tyrosine hydroxylase gene therapy of the brain with Trojan horse liposomes
Authors: Xia Chun‐Fang , Chu Chun , Li Jianyi , Wang Yuntao , Zhang Yun , Boado Ruben J. , Pardridge William M. 
Published: May 2007

AbstractBackgroundThe present study examines whether chromosomal derived forms of therapeutic genes can be delivered to brain following intravenous administration. The brain expression of a rat tyrosine hydroxylase (TH) cDNA is compared to the brain expression of a plasmid DNA encoding the 18 kb rat TH gene.MethodsTH gene expression is measured in cell culture and in vivo in brain in experimental Parkinson's disease (PD). A total of four eukaryotic expression plasmids encoding rat TH were engineered wherein the size of the TH expression cassette ranged from 1.5 kb, in the case of the cDNA form of the gene, to 17.5 kb, in the case of the largest size genomic construct. The TH expression plasmids were delivered to either cultured cells or to rat brain in vivo with Trojan horse liposomes (THLs), which target the non‐viral plasmid DNA to cells via cell membrane receptors.ResultsThe pattern of TH gene expression in cell culture and in vivo was similar: the cDNA form of the TH gene was fast‐acting with short duration of action, and the genomic form of the TH gene was slow‐acting with longer duration of action. The most sustained replacement of striatal TH enzyme activity in experimental PD was produced by combination gene therapy where both the cDNA and the genomic forms of the TH gene were administered simultaneously.ConclusionsEukaryotic expression plasmids encoding genomic forms of therapeutic genes, as large as 18 kb, can be successfully incorporated in THLs and delivered to brain following intravenous administration. Copyright © 2007 John Wiley & Sons, Ltd.
The application of electrospray ionization coupled to ultrahigh resolution mass spectrometry for the molecular characterization of natural organic matter
Authors: Sleighter Rachel L. , Hatcher Patrick G. 
Published: May 2007

AbstractMass spectrometry has recently played a key role in the understanding of natural organic matter (NOM) by providing molecular‐level details about its composition. NOM, a complex assemblage of organic molecules present in natural waters and soils/sediments, has the ability to bind and transport anthropogenic materials. An improved understanding of its composition is crucial in order to understand how pollutants interact with NOM and how NOM cycles through global carbon cycles. In the past, low‐resolution (>3000) mass analyzers have offered some insights into the structure of NOM, but emerging ultrahigh resolution (>200 000) techniques such as electrospray ionization (ESI) coupled to Fourier transform ion cyclotron resonance mass spectrometry (FT‐ICR‐MS) have significantly advanced our knowledge of NOM chemistry. Here, a review of the recent literature on the advancements of NOM characterization and the applications of mass spectrometry to this central task is presented. Various methods for the analysis and display of the extremely complex mass spectra, such as the van Krevelen diagram and Kendrick mass defect analysis, are discussed. We also review tandem mass spectrometry techniques employed to gain structural information about NOM components. Finally, we show how ESI‐FT‐ICR‐MS has been applied to examine specific issues that are important to the NOM scientific community, such as NOM reactivity, transport and fate, degradation, and existence of components, which are indicators of NOM origin. In general, ultrahigh resolution provided by FT‐ICR‐MS is essential for the complete separation of the thousands of peaks present in the complex NOM mixture and will clearly lead to additional future advancements in the areas of aquatic, soil, and analytical chemistry. Copyright © 2007 John Wiley & Sons, Ltd.
Gastrojejunostomy versus stent placement in patients with malignant gastric outlet obstruction: a comparison in 95 patients
Authors: Jeurnink S.M. , Steyerberg E.W. , Hof G. van 'T , van Eijck C.H.J. , Kuipers E.J. , Siersema P.D. 
Published: May 2007

AbstractAimGastrojejunostomy (GJJ) and duodenal stent placement are the most commonly used palliative treatment modalities for gastric outlet obstruction (GOO). In this retrospective study, we compared GJJ and stent placement with regard to medical effects.MethodsMedical records of 95 patients who had undergone palliative treatment between 1994 and 2006 in a Dutch university hospital, were reviewed. Study outcomes were improvement of food intake, complications, persistent and recurrent symptoms, re‐interventions, hospital stay, and survival.ResultsFifty‐three patients were referred for duodenal stent placement and 42 patients underwent GJJ. There were no differences in technical and clinical success and the incidence of minor and early major complications and survival. Food intake improved more rapidly after stent placement than GJJ (P = 0.01). The time to late major complications, recurrent obstructive symptoms and re‐intervention was significantly shorter after stent placement than GJJ (P = 0.004, 0.002, and 0.004, respectively). Hospital stay was also shorter after stent placement than GJJ (P < 0.001).ConclusionThese findings suggest that stent placement is associated with better short‐term outcomes and GJJ with better long‐term outcomes. A large randomized controlled trial is however needed to systematically compare stent placement with GJJ with regard to medical effects, quality of life and costs. J. Surg. Oncol. 2007;96:389–396. © 2007 Wiley‐Liss, Inc.
Uniqueness Theorems for the Frankl‐Morawetz Problem in R2 for a Class of Nonlinear Equations of Mixed Type
Authors: Aziz A. K. , Schneider M. 
Published: May 2007

We consider the question of uniqueness for the solutions of the boundary value problem
urn:x-wiley:0025584X:mana19861250115:equation:mana19861250115-math-0001 where sgn K(y) = sgn y and Γ0 and Γ1 are parts of the boundary of a bounded simply connected region G in R2. G is bounded for y > 0 by a piecewise smooth curve Γ0 which intersects the line y = 0 at A (– 1, 0) and B(0, 0). For y < 0 G is bounded by a piecewise smooth curve Γ1 through A, which meets the characteristic of (1) issued from B at point C, and by the curve Γ2 which consists of the portion CB of the characteristic through B.Using energy‐integral considerations, we give sufficient conditions for the uniqueness of solutions to boundary value problem (1).
On Some Inequalities for Poisson Networks
Authors: Mecke J. 
Published: May 2007

AbstractA stationary Poisson hyperplane process in Rd induces a random network of (d‐2)‐flats, each of which is the intersection of two hyperplanes of the process. It is known that the intensity of the induced (d‐2)‐flat process divided by the square of the intensity of the original hyperplane process is maximal in the isotropic case. An integral‐geometric formula for elliptic spaces is presented, from which the mentioned extremum property and related inequalities for superpositions of stationary Poisson hyperplane processes are derived.
Spinoren in statischen Raum‐Zeiten
Authors: Illge Reinhard 
Published: May 2007

AbstractIn this paper, a spinor algebra and analysis adapted to static space‐times is presented. Suitable SU(2)‐bases are choosen in spinor space and it is shown, how these bases determine orthogonal systems in (three‐dimensional) space. Some theorems on the curvature spinors of static space‐times are proved by the help of the calculus of the connection spinors. The internal structure of the WEYL spinor as well as its connection with the RICCI tensor of the underlying (three‐dimensional) space are examined. The presented calculus allows the computation of the NEWMAN‐PENROSE spin coefficients and the canonical normal 1‐spinors of the WEYL spinor with a relatively small expense, which is demonstrated on a sequence of examples.
Lösungsverzweigung bei analytischen Gleichungen mit Fredholm‐Operator von Index Null
Authors: Heinz Günter 
Published: May 2007

AbstractThe bifurcation of a solution of the equationf(x, λ) = 0 at the point (x0, λ0) is investigated. In the case thatB: = –fx(x0, λ0) is a FREDHOLM operator by the method of LJAPUNOV/SCHMIDT the original equation is equivalent to a system consisting of a locally uniquely solvable equation and an equation in a finit dimensional subspace, the so‐called bifurcation equation. For analytical/recursion formulas are deduced to determine the locally unique solution. In the case of FREDHOLM operatorsBwith index zero practicable criteria are given for the applicability of a theorem of IZE being a generalization of a well known theorem of KRASNOSELSKIJ.
A Sheaf Property for Excessive Functions of Right Processes
Authors: Steffens J. 
Published: May 2007

AbstractOn a state space of a right process a facial topology depending upon some RAY compactification is introduced. With respect to this facial topology the right process is shown to exhibit a nice path behaviour. Furthermore, conditions are given which permit a local characterization of excessive functions with respect to the facial topology. This result gives a partial answer to an open question within the potential theory of H‐cones.
Totally Incomparable Banach Spaces and Three‐Space Banach Space Ideals
Authors: Alvarez Teresa , Gonzalez Manuel , Onieva Victor M. 
Published: May 2007

AbstractIn this paper we present four methods to generate three‐space BANACH space ideals. They are based on the concept of total incomparability of H. P. ROSENTHAL and on a dual concept, total coincomparability, which is here introduced. We use the assertion that the sum of two totally incomparable closed subspaces of a BANACH space is norm‐closed, which is shown by means of an easier and more natural proof than that of ROSENTHAL [10], and an analogous property about the total coincomparability. Several well‐known ideals are obtained with the above methods, and so they are three‐space ideals.
Polyhedron Theorems for Non‐Smooth Cell Complexes
Authors: Zähle M. 
Published: May 2007

AbstractResults like EULER'S polyhedron theorem or relations for the EULER characteristic are extended to signed curvature measures of p‐dimensional topological polyhedra in Rd. The cells are assumed to be finite unions of compact manifolds with boundary and positive reach.By means of generalized unit normal bundles and generalized principal curvatures provided with a certain set additive index function the global problems can be reduced to local variants.
The Extended Probability Generating Functional, with Application to Mixing Properties of Cluster Point Processes
Authors: Daley D. J. , Vere‐Jones D. 
Published: May 2007

AbstractAn extended probability generating functional (p. g. fl.)\documentclass{article}\pagestyle{empty}\begin{document}$ \bar{G}[h]\; = \;E\;\left({\exp \;\int\limits_x {\log } \;h(x)\; \times \;N(dx)} \right) $\end{document}is well‐defined for any point processNon the complete separable metric space χ over the space V̄0of measurable functionsh: χ → (0, 1) such that infxcHh(x) > 0. The distribution ofNis determined uniquely by the p.g.fl.G[h] ≡ Ḡ[h] over the smaller spaceV0of functionshε V̄0for which 1 —hhas bounded support. Continuity results for Ḡ[·] involving pointwise convergent sequences {hn}V0or V̄0or V̄ ≡ {measurableh: χ → [0, 1]} orV= {hε V̄: 1 —hhas bounded support} are reviewed, and used in furnishing a complete p. g. fl. proof of the mixing property of certain stationary cluster processes.
Microwave‐assisted proteomics
Authors: Lill Jennie R. , Ingle Elizabeth S. , Liu Peter S. , Pham Victoria , Sandoval Wendy N. 
Published: May 2007

AbstractState‐of‐the‐art proteomic analysis has recently undergone a rapid evolution; with more high‐throughput analytical instrumentation and informatic tools available, sample preparation is becoming one of the rate‐limiting steps in protein characterization workflows. Recently several protocols have appeared in the literature that employ microwave irradiation as a tool for the preparation of biological samples for subsequent mass spectrometric characterization. Techniques for microwave‐assisted bio‐catalyzed reactions (including sample reduction and alkylation, enzymatic and chemical digestion, removal and analysis of post‐translational modifications and characterization of enzymes and protein‐interaction sites) are described. This review summarizes the various approaches undertaken, instrumentation employed, and reduction in overall experimental time observed when microwave assistance is applied. © 2007 Wiley Periodicals, Inc., Mass Spec Rev 26:657–671, 2007
Reprogramming events of mammalian somatic cells induced by Xenopus laevis egg extracts
Authors: Miyamoto Kei , Furusawa Tadashi , Ohnuki Mari , Goel Sandeep , Tokunaga Tomoyuki , Minami Naojiro , Yamada Masayasu , Ohsumi Keita , Imai Hiroshi 
Published: May 2007

AbstractIt is known that differentiated cells can be reprogrammed to an undifferentiated state in oocyte cytoplasm after nuclear transfer. Recently, some reports suggested that Xenopus egg extracts have the ability to reprogram mammalian somatic cells. Reprogramming events of mammalian cells after Xenopus egg extract treatment and after cell culture of extract‐treated cells have not been elucidated. In this experiment, we examined reprogramming events in reversibly permeabilized or nonpermeabilized porcine fibroblast cells after Xenopus egg extract treatment. The Xenopus egg‐specific histone B4 was assembled on porcine chromatin and nuclear lamin LIII was incorporated into nuclei. Deacetylation of histone H3 at lysine 9 in extract‐treated cells was detected in nonpermeabilized cells, suggesting that a part of reprogramming may be induced even in nonpermeabilized cells. Following culture of extract‐treated cells, the cells began to express the pluripotent marker genes such as POU5F1 (OCT4) and SOX2 and to form colonies. Reactivation of the OCT4 gene in extract‐treated cells was also confirmed in bovine fibroblasts transformed with an OCT4‐EGFP construct. These results suggest that nuclei of mammalian cells can be partially reprogrammed to an embryonic state by Xenopus egg extracts and the remodeled cells partly dedifferentiate after cell culture. A system using egg extracts may be useful for understanding the mechanisms and processes of dedifferentiation and reprogramming of mammalian somatic cells after nuclear transfer. Mol. Reprod. Dev. 74: 1268–1277, 2007. © 2007 Wiley‐Liss, Inc.
Differential but beneficial effect of phytohemagglutinin on efficiency of in vitro porcine embryo production by somatic cell nuclear transfer or in vitro fertilization
Authors: Gupta Mukesh Kumar , Uhm Sang Jun , Lee Hoon Taek 
Published: May 2007

AbstractIn vitro production of porcine embryos has become routine in most laboratories but the yield and quality of the resultant blastocysts remains sub‐optimal. Phytohemagglutinin (PHA) is an N‐acetylgalactosamine/galactose sugar‐specific lectin with a wide variety of biological activities including mitogenesis, mediation of cell recognition and agglutination of cells. This study was therefore, designed to investigate the effect of PHA on the preimplantation development and quality of parthenogenetic, somatic cell nuclear transferred (SCNT) and in vitro fertilized (IVF) porcine embryos cultured in the absence or presence of PHA. Analysis showed that, supplementation of PHA significantly improved the blastocyst rate of parthenogenetic (70.6 ± 0.2 vs. 51.4 ± 0.6%) and SCNT (27.7 ± 1.7 vs. 12.5 ± 0.3%) embryos but not IVF embryos (25.0 ± 14.3 vs. 20.1 ± 12.7%). Nonetheless, PHA‐treated blastocysts had higher hatching ability and contained higher cell number than control blastocysts in all the groups (P < 0.05). TUNEL labeling revealed that blastocysts cultured in the presence of PHA were less predisposed to biochemical apoptosis and showed lower indices of TUNEL, fragmentation and total apoptosis than those cultured in the absence of PHA (P < 0.05). Real time qRT‐PCR analysis of parthenogenetic blastocysts revealed that PHA decreased the expression ratio of BAX:BCL‐xL transcripts. Therefore, our study suggests that PHA improves the blastocyst yield and quality by enhancing blastocyst expansion, hatching and total cell number and decreasing the apoptosis. However, PHA has a differential effect on development rate of IVF derived embryos. These results may represent an approach towards achieving better preimplantation development of porcine embryos in vitro. Mol. Reprod. Dev. 74: 1557–1567, 2007. © 2007 Wiley‐Liss, Inc.