bluuebunny/crossref_metadata_2025 huggingface dataset:
Ba6Si6N10O2(CN2) – A Nitridosilicate with a NPO‐Zeolite Structure Type Containing Carbodiimide Ions
Authors: Pagano Sandro , Oeckler Oliver , Schröder Thorsten , Schnick Wolfgang
Published: May 2009

A new precursor approach leading to NPO‐zeolite analogous nitridosilicates with cavities containing carbodiimide ions is presented. The reaction of amorphous “Si(CN2)2” and barium in liquid sodium afforded Ba6Si6N10O2(CN2) as yellow crystals. The structure is a rare example of the NPO‐zeolite framework type and the first nitridosilicate incorporating carbodiimide ions. The partially ordered integration of carbodiimide moieties in the channels leads to the formation of a superstructure (P$\bar {6}$, no. 174, a = 16.255(2), c = 5.4690(11) Å, Z = 3, R1 = 0.0299, 2139 data, 100 parameters) and merohedral twinning. A comprehensive structure solution is presented, taking all possible ordering variants and twin laws into account.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
Synthesis and Characterisation of Low‐Coordinate Transition‐Metal Complexes Stabilised by Sterically Demanding Carbazolido Ligands
Authors: Ashley Andrew E. , Cowley Andrew R. , Green Jennifer C. , Johnston David R. , Watkin David J. , Kays Deborah L.
Published: May 2009

The synthesis and characterisation of the homoleptic transition‐metal complexes (1,8‐Ph2‐3,6‐Me2C12H4N)2M (M = Cr, 1; M = Mn, 2; M = Fe, 3; M = Co, 4) are described. Formally two‐coordinate, the solid‐state structures of 2 and 3 reveal near‐linear N–M–N geometries and significant bending of the flanking phenyl groups towards the metal centres. Solid‐state magnetic measurements reveal high spin metal centres in 1–4; computational calculations for the model complexes M(1,8‐Ph2C12H6N)2 (M = Cr–Co) confirm the presence of high‐spin configurations within these bis‐carbazolido compounds and an orbital ordering of δ < π < σ for these systems.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
Templated Synthesis and Site‐Selective Conversion of Completely Nonsymmetrical Bis‐Metallosalphen Complexes
Authors: Castilla Ana M. , Curreli Simona , Carretero Nina M. , Escudero‐Adán Eduardo C. , Benet‐Buchholz Jordi , Kleij Arjan W.
Published: May 2009

A metal‐templated, stepwise approach toward fully nonsymmetrical bis‐metallosalphen complexes is described. The synthesis comprises the selective monometalation of diimine precursors and subsequent introduction of the second metal ion by using various salicylaldehyde reagents. In the case of a bis‐Zn(salen) derivative, the presence of different peripheral substituents on the two metallosalen units gave rise to a large difference in kinetic stability, which was used to selectively demetalate the more labile site. This result shows promise for the preparation of heterobimetallic structures. The first X‐ray molecular structure of a completely nonsymmetrical bis‐salphen complex is also reported. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
Ternary Porphyrinato HfIV and ZrIV Polyoxometalate Complexes
Authors: Falber Alexander , Burton‐Pye Benjamin P. , Radivojevic Ivana , Todaro Louis , Saleh Raihan , Francesconi Lynn C. , Drain Charles Michael
Published: May 2009

We report a facile, high‐yield synthesis and characterization of discrete, ternary porphyrin–metal–polyoxometalate (por–M–POM) complexes where a group IV transition‐metal ion is bound both to the porphyrin core and to the lacunary site of a Keggin POM, PW11O39–7. The remarkably robust complexes exploit the fact that HfIV and ZrIV are 7–8 coordinate and reside outside the plane of the porphyrin macrocycle, thus enabling the simultaneous coordination to meso‐tetraphenylporphyrin (tpp) or meso‐tetra(4‐pyridyl)porphyrin (tpyp) and to the defect site in the Keggin framework. The physical properties of the (tpp)Hf(PW11O39)[tba]5, (tpyp)Hf(PW11O39)[tba]5, and (tpp)Zr(PW11O39)[tba]5 complexes are similar because the metal ions have similar oxidation states and coordination chemistry. This architecture couples the photonic properties of the porphyrin to the POM because the metal ion is incorporated into both frameworks. Thus, the ternary complexes can serve as a basis for the characterization of HfIV and ZrIV porphyrins bound to oxide surfaces through the group IV metal ions. The Hf(por) and Zr(por) bind strongly to TiO2 nanoparticles and indium tin oxide (ITO) surfaces, but significantly less binds to crystalline SiO2 or TiO2 surfaces. Together, the strong binding of the metalloporphyrins to the POM, nanoparticles, and the ITO surfaces, and paucity of binding to crystalline surfaces, suggests that the three‐to‐four open coordination sites on the Hf(por) and Zr(por) are predominantly bound at surface defect sites. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
Studies towards the Total Synthesis of (–)‐Caulerpenynol, a Toxic Sesquiterpenoid of the Green Seaweed Caulerpa taxifolia
Authors: Commeiras Laurent , Thibonnet Jérôme , Parrain Jean‐Luc
Published: May 2009

The first diastereoselective synthesis of the antimicrobial and cytotoxic agent (–)‐caulerpenynol (2) was achieved in relatively few steps from commercially available (S)‐malic acid. Highlights of this synthesis include the nonracemization of the sensitive α‐hydroxy ketone moiety and the proper choice of the protecting groups for critical last deprotection step. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
Synthesis of 2‐Azetidinones Incorporating Carbenechromium(0) Moieties and Their Use in the Preparation of Penicillin‐ and Cephalosporin‐Containing Peptides
Authors: Sierra Miguel A. , Rodríguez‐Fernández Mamen , Casarrubios Luis , Gómez‐Gallego Mar , Mancheño María José
Published: May 2009

A series of β‐lactams incorporating a carbenechromium(0) moiety in their structures have been prepared. These stable compounds are suitable precursors for the preparation of α‐amino esters, dipeptides and tripeptides tethered to a 2‐azetidinone ring. The methodology has also been extended to the synthesis of carbenechromium(0) complexes of penicillin G, cephalosporin and 6‐aminopenicillanic acid. All these new metalla‐β‐lactam derivatives are stable compounds and can be transformed into antibiotic structures with tripeptide side‐chains. The methodology relies on the compatibility of the β‐lactam ring with the carbenechromium(0) moiety and on the stability of the four‐membered ring during the photocarbonylation of the resulting chromium complex.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
An Easy and Multigram‐Scale Synthesis of Versatile AA‐ and AB‐Type m‐Terphenylenes as Building Blocks for Kinked Polyphenylenes
Authors: Kissel Patrick , Breitler Simon , Reinmüller Viktoria , Lanz Patrick , Federer Lukas , Schlüter A. Dieter , Sakamoto Junji
Published: May 2009

A set of m‐terphenylenes having a readily functionalizable hydroxy group as well as either symmetric AA‐type or unsymmetric AB‐type halide termini have been prepared on a several‐gram scale. The synthesis was carried out on the basis of the Suzuki–Miyaura cross‐coupling in combination with a TMS masking strategy. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
Long open tubular molecule imprinted polymer capillary columns with excellent separation efficiencies in chiral and non‐chiral separation by capillary electrochromatography
Authors: Zaidi Shabi Abbas , Cheong Won Jo
Published: May 2009

Long open tubular S‐ketoprofen molecule imprinted polymer capillary columns (1, 2, and 3 m) with the number of theoretical plates (N) over a million for both chrial and non‐chiral CEC separation have been prepared. Chiral separation of racemic ketoprofen and non‐chiral separation of other profen drugs have been achieved with excellent efficiency and good selectivity after optimization of the chromatographic conditions. The N value per column was not proportional to the effective column length although it was increased with the column length. This study has shown the outstanding prospect of the strategy to form a porous and rugged open tubular layer in a moderately long (1 m) capillary column in molecule imprinted polymer‐CEC studies.
Electrophoretic behaviors of human hepatoma HepG2 cells
Authors: Hsu Jyh‐Ping , Ku Ming‐Hong , Yang Li‐Chia , Lu Jui‐Nan , Young Tai‐Horng , Tseng Shiojenn
Published: May 2009

The electrophoretic mobility of HepG2 cells was measured and a charge‐regulated model was proposed to simulate the results obtained. Here, a cell was simulated by a rigid core and an ion‐penetrable membrane layer containing both acidic and basic functional groups. The influences of the key parameters, including the pH, the ionic strength, the thickness of the membrane layer of a cell, the density and the dissociation constant of the dissociable functional groups in the membrane layer, and the binding constant of divalent cations on the electrophoretic mobility of a cell were investigated. In particular, the role of the buffer used in the experiment was discussed; this effect was neglected in almost all the relevant theoretical analyses in the literature. We showed that the binding ability of divalent cations to the dissociated functional groups in the membrane layer of a cell ranks as Ca2+>Mg2+>hexamethonium.
Enhanced detection in capillary electrophoresis: Example determination of serum mycophenolic acid
Authors: Shihabi Zak K.
Published: May 2009

In order to overcome the poor absorbency detection limits in CE, two simple strategies were combined to increase the amount of the sample injected: a long capillary to hold extra sample while leaving adequate room for the separation and acetonitrile stacking, which concentrated the sample based on transient pseudo‐ITP. The combination of these two strategies yielded sensitivity comparable or better than that of the HPLC with good separation and with better theoretical plate number. The analysis of mycophenolic acid, a common immunosuppressant drug, in serum was used here as an example to illustrate this enhanced detection and its applicability to therapeutic drug monitoring. Acetonitrile was used to remove serum proteins followed by direct injection filling 5–21% of the capillary volume and separation in a borate buffer. The overall CE method compared well to an assay by HPLC as far as sample preparation, correlation coefficient, and especially sensitivity of detection.
Capillary electrophoretic study of small, highly sulfated, non‐sugar molecules interacting with antithrombin
Authors: Liang Aiye , Raghuraman Arjun , Desai Umesh R.
Published: May 2009

Affinity CE (ACE) was used to study interactions of small, highly sulfated, aromatic molecules with antithrombin (AT). The high charge density of the small molecules induces differential migration of the complex resulting in a versatile method of assessing binding affinities, nature of interactions and site of binding on the inhibitor. Scatchard analysis of the interaction of three tetrahydroisoquinoline‐based polysulfated molecules with AT results in monophasic profiles with affinities in the range of 40–60 μM in 20 mM sodium phosphate buffer, pH 7.4. For a pentasulfated molecule, a biphasic profile with affinities of 4.7 and 30 μM was observed. Measurement of KD as a function of ionic strength of the medium indicated that ionic and non‐ionic forces contribute 2.4 and 1.9 kcal/mol, respectively, at pH 7.4 and 100 mM NaCl. Competitive binding studies showed that the tetrahydroisoquinoline‐based molecules do not compete with a high‐affinity heparin pentasaccharide. In contrast, the affinity of these tetrahydroisoquinoline derivatives decreases dramatically in the presence of an extended heparin‐binding site ligand. Overall, ACE analysis of small, sulfated aromatic molecules interacting with AT is relatively easy and obviates the need for an external signal, e.g. fluorescence, for monitoring the interaction. In addition to affording biochemical knowledge, the small sample requirement and fast analysis time of ACE could be particularly advantageous for high‐throughput screening of potential anticoagulants.
Insulator‐based dielectrophoretic separation of small particles in a sawtooth channel
Authors: Chen Kang Ping , Pacheco Jose Rafael , Hayes Mark. A. , Staton Sarah J. R.
Published: May 2009

Insulator‐based dielectrophoretic separation of small particles in a sawtooth channel is studied in the limit of dilute concentration. Pathlines for the movements of infinitesimal particles are constructed and the geometric changes of these pathlines are used to establish the criterion for blocking and trapping particles with different physical properties. The sharp corners of the sawtooth channel create much stronger dielectrophoretic force than channels with smooth corners for blocking particle movements. Particle blocking and trapping depend on particle properties and the geometry of the device. It is shown that once the channel geometric aspect ratios are specified, the blocking criterion depends on only a single dimensionless parameter C defined in terms of the particle mobility ratio (dielectrophoretic versus electrokinetic), the applied voltage and the spacing between the teeth. Selective blocking and trapping of particles can be realized by varying the geometry of the channel progressively. High‐resolution separation can be achieved by tuning the differential in the parameter C to a desired level.
Proteome alteration of early‐stage differentiation of mouse embryonic stem cells into hepatocyte‐like cells
Authors: Li Yan , Kang Xiaonan , Guo Kun , Li Xuefei , Gao Dongmei , Cui Jiefeng , Sun Lu , Yang Pengyuan , Liu Yinkun
Published: May 2009

To explore the molecular basis of inducible differentiation of embryonic stem cells into hepatocyte‐like cells, a proteomic strategy was utilized to examine the global protein expression alterations after early‐stage differentiation of a mouse D3 embryonic stem (ES) cell line along hepatic lineage. The undifferentiated D3 cells were treated stepwise with combinations of defined chemicals and growth factors. The differentiated cells were identified by hepatocyte‐like morphology, expressed liver‐specific markers as well as the evidence of glycogen storage. The subsequent proteomic separation and identification were performed with 2‐DE followed by MALDI‐TOF‐MS/MS analysis. Of the 119 differentially displayed protein spots analyzed, 90 spots presenting 64 distinct proteins were finally identified. The interested protein expressions were validated by Western blotting such as albumin and cytokeratin‐8. Bioinformatic annotations indicated that this set of proteins was enriched with transcription, translation regulation and protein processing, energy/metabolism and chaperon functions. A part of them had been found to be involved in the differentiation of mouse ES cells. Interestingly, approximately 40% of these proteins had been previously reported as being dysregulated in hepatocellular carcinoma. It suggested that these changed proteins may be candidate regulators of ES cell differentiation, some of them may be specific to hepatic differentiation.
Analysing risk attitudes to time
Authors: Oliver Adam , Cookson Richard
Published: May 2009

The assumption of risk neutrality over discounted life years underlies the standard QALY model of individual preferences over health outcomes, and is thus implicitly assumed by NICE and other health technology advisory bodies worldwide. The primary objective of this article is to report a study to test the assumption in a convenience sample of 30 respondents with use of the probability equivalence version of the standard gamble. The results indicate considerable risk aversion over life years, and therefore call into question the standard assumption of risk neutrality in practical cost‐utility analyses (CUA). A secondary objective is to observe whether risk aversion can be reduced through the use of the lottery equivalents method, under the hypothesis that the gambling effect can be lessened with this instrument. In a separate convenience sample of 40 respondents, however, the observed level of risk aversion was at least that seen in the standard gamble. Further research is warranted to ascertain whether risk aversion over discounted life years is a generalisable concern. Copyright © 2009 John Wiley & Sons, Ltd.
Reuse of return flow from wetland paddy to supplement industrial water needs
Authors: Chang Yu‐Chuan , Lin Wei‐Taw , Hsu Nien‐Sheng , Kan Chun‐E.
Published: May 2009

In this paper a water reuse system in wetland paddy was simulated to supply water for industry. A two‐tank model was employed to represent the return flow of the wetland paddy. Historical data were used for parameter estimation and model verification. The simulation results show that the return flow from wetland paddy has potential benefit for water reuse. Under a series of water management strategies, about 87% of the total volume of return flow can supplement industrial water needs which decreases local water needs by about 22.4%. The strategies include increasing the volume of return flow through deepwater management practices, stabilizing the reuse of return flow in the purification unit through the regulated large‐capacity pond downstream, and harmonizing the supply‐and‐demand gap through a suitable cultivated paddy scale of the crop rotation pattern. Apart from meeting water needs at peak demand periods, the reuse of return flow is a quick‐response water supply solution. Accompanied by the series of water management strategies, water reliability and crop security can be achieved at the same time. Copyright © 2009 John Wiley & Sons, Ltd.
Effect of Filtered Cultures of Flagellate Ochromonas sp. on Colony Formation in Microcystis aeruginosa
Authors: Yang Zhen , Kong Fanxiang , Zhang Min , Yang Zhou , Yu Yang , Qian Shanqin
Published: May 2009

The possible effect of filtered cultures of flagellate Ochromonas sp. on colony formation in M. aeruginosa was investigated in this paper. The results show that filtered cultures of flagellates fed with M. aeruginosa could induce colony formation in M. aeruginosa. Furthermore, induction strength is clearly dependent on the concentration of flagellates and filtered cultures. However, no colonial M. aeruginosa was found in the treatments of filtered cultures of flagellates fed with Microcystis wesenbergii, filtered cultures of flagellate fed with Chlorella pyrenoidosa, and algae homogenates. This suggests that infochemicals released from flagellates fed with M. aeruginosa may be a trigger for colony formation in M. aeruginosa. The clearance rates of flagellates on algae were markedly decreased when they were cultivated with induced colonial M. aeruginosa. These indicate that colony formation in M. aeruginosa is a predator‐induced defense which could reduce predation risk from flagellates (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Characterization and tribology of PEG‐like coatings on UHMWPE for total hip replacements
Authors: Kane Sheryl R. , Ashby Paul D. , Pruitt Lisa A.
Published: May 2009

A crosslinked hydrogel coating similar to poly(ethylene glycol) (PEG) was covalently bonded to the surface of ultrahigh molecular weight polyethylene (UHMWPE) to improve the lubricity and wear resistance of the UHWMPE for use in total joint replacements. The chemistry, hydrophilicity, and protein adsorption resistance of the coatings were determined, and the wear behavior of the PEG‐like coating was examined by two methods: pin‐on‐disk tribometry to evaluate macroscale behavior, and atomic force microscopy (AFM) to simulate asperity wear. As expected, the coating was found to be highly PEG‐like, with ∼83% ether content by x‐ray photoelectron spectroscopy and more hydrophilic and resistant to protein adsorption than uncoated UHMWPE. Pin‐on‐disk testing showed that the PEG‐like coating could survive 3 MPa of contact pressure, comparable to that experienced by total hip replacements. AFM nanoscratching experiments uncovered three damage mechanisms for the coatings: adhesion/microfracture, pure adhesion, and delamination. The latter two mechanisms appear to correlate well with wear patterns induced by pin‐on‐disk testing and evaluated by attenuated total reflection Fourier transform infrared spectroscopy mapping. Understanding the mechanisms by which the PEG‐like coatings wear is critical for improving the behavior of subsequent generations of wear‐resistant hydrogel coatings. © 2009 Wiley Periodicals, Inc. J Biomed Mater Res 2010
Dendritic cell responses to self‐assembled monolayers of defined chemistries
Authors: Shankar Sucharita P. , Petrie Timothy A. , García Andrés J. , Babensee Julia E.
Published: May 2009

Biomaterial contact triggers dendritic cell (DC) maturation, to an extent depending on the biomaterial, ultimately enhancing an immune response toward associated antigens, implying a role for biomaterials as adjuvants. Self‐assembled monolayers (SAM) of alkanethiols on titanium/gold‐coated surfaces presenting different chemistries were used to study effects of biomaterial surface chemistry on DC maturation. Although DCs treated with OH, COOH, or NH2SAMs showed modest maturation, those treated with CH3SAMs were least mature, all based on cytospins, allostimulatory capacity, or maturation marker expression. Surprisingly, DCs treated with CH3SAMs secreted highest levels of proinflammatory tumor necrosis factor‐α (TNF‐α) or interleukin‐6 (IL‐6) but were least mature. Secretion of anti‐inflammatory mediators by DCs treated with CH3SAMs was not responsible for mitigating DC maturation under these conditions. Interestingly, elevated levels of apoptotic markers were measured associated with DCs and T cells upon CH3SAMs contact. Since phagocytosis of apoptotic DCs has strong immunosuppressive effects on DCs, more apoptotic DCs on CH3SAMs may account for lower DC maturation. Finally, higher expression of cytotoxic T lymphocyte associated antigen receptor‐4 (CTLA‐4) on T cells may imply a mechanism of T cell inhibition on CH3SAMs. © 2009 Wiley Periodicals, Inc. J Biomed Mater Res 2010
Microcomputed tomography and microfinite element modeling for evaluating polymer scaffolds architecture and their mechanical properties
Authors: Alberich‐Bayarri Angel , Moratal David , Ivirico Jorge L. Escobar , Hernández José C. Rodríguez , Vallés‐Lluch Ana , Martí‐Bonmatí Luis , Estellés Jorge Más , Mano Joao F. , Pradas Manuel Monleón , Ribelles José L. Gómez , Salmerón‐Sánchez Manuel
Published: May 2009

Detailed knowledge of the porous architecture of synthetic scaffolds for tissue engineering, their mechanical properties, and their interrelationship was obtained in a nondestructive manner. Image analysis of microcomputed tomography (μCT) sections of different scaffolds was done. The three‐dimensional (3D) reconstruction of the scaffold allows one to quantify scaffold porosity, including pore size, pore distribution, and struts' thickness. The porous morphology and porosity as calculated from μCT by image analysis agrees with that obtained experimentally by scanning electron microscopy and physically measured porosity, respectively. Furthermore, the mechanical properties of the scaffold were evaluated by making use of finite element modeling (FEM) in which the compression stress–strain test is simulated on the 3D structure reconstructed from the μCT sections. Elastic modulus as calculated from FEM is in agreement with those obtained from the stress–strain experimental test. The method was applied on qualitatively different porous structures (interconnected channels and spheres) with different chemical compositions (that lead to different elastic modulus of the base material) suitable for tissue regeneration. The elastic properties of the constructs are explained on the basis of the FEM model that supports the main mechanical conclusion of the experimental results: the elastic modulus does not depend on the geometric characteristics of the pore (pore size, interconnection throat size) but only on the total porosity of the scaffold. © 2009 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2009
Effect of black tea on histological and immunohistochemical changes in pancreatic tissues of normal and streptozotocin‐induced diabetic mice (Mus musculus)
Authors: Manikandan Ramar , Sundaram Ramalingam , Thiagarajan Raman , Sivakumar Mullaivanam Ramasamy , Meiyalagan Velayutham , Arumugam Munusamy
Published: May 2009

The aim of the present study is to evaluate the effect of hot water extract of black tea in regenerating β cells in streptozotocin‐induced diabetic mice. Light microscopic examination of pancreatic sections of streptozotocin‐induced diabetic mice showed the acinar cells to be small, shrunken, and with deteriorated β cells. The dose of streptozotocin not only altered the function of β cells but also damaged the acinar region. The changes in acinar cells were coarsening of endoplasmic reticulation suggesting alteration in their secretory function. The control pancreatic tissue showed well‐defined granulated islets and dark β cells when stained with chrome hematoxylin and phloxine. Interestingly, pancreatic sections of diabetic mice fed with black‐tea extract showed regeneration of β cells and acinar region appeared normal with increased numbers of β cells. To understand the probable mechanism of action of black‐tea extract, we analyzed inducible nitric oxide synthase (iNOS) expression by immunohistochemistry and the results showed an increased iNOS levels in streptozotocin‐induced diabetic pancreas, and such high iNOS levels were inhibited in black‐tea extract treated mice. According to histological results obtained, it can be concluded that the black‐tea extract helps in regeneration of damaged pancreas and protects pancreatic β cells by its antioxidant action against nitrosative stress in streptozotocin‐induced diabetes. Microsc. Res. Tech., 2009. © 2009 Wiley‐Liss, Inc.
Gene expression analysis of immunostained endothelial cells isolated from formaldehyde‐fixated paraffin embedded tumors using laser capture microdissection—A technical report
Authors: Kaneko Tomoatsu , Okiji Takashi , Kaneko Reika , Suda Hideaki , Nör Jacques E.
Published: May 2009

Laser capture microdissection (LCM) allows microscopic procurement of specific cell types from tissue sections that can then be used for gene expression analysis. In conventional LCM, frozen tissues stained with hematoxylin are normally used to the molecular analysis. Recent studies suggested that it is possible to carry out gene expression analysis of formaldehyde‐fixated paraffin embedded (FFPE) tissues that were stained with hematoxylin. However, it is still unclear if quantitative gene expression analyses can be performed from LCM cells from FFPE tissues that were subjected to immunostaining to enhance identification of target cells. In this proof‐of‐principle study, we analyzed by reverse transcription‐PCR (RT‐PCR) and real time PCR the expression of genes in factor VIII immunostained human endothelial cells that were dissected from FFPE tissues by LCM. We observed that immunostaining should be performed at 4°C to preserve the mRNA from the cells. The expression of Bcl‐2 in the endothelial cells was evaluated by RT‐PCR and by real time PCR. Glyceraldehyde‐3‐phosphate dehydrogenase and 18S were used as house keeping genes for RT‐PCR and real time PCR, respectively. This report unveils a method for quantitative gene expression analysis in cells that were identified by immunostaining and retrieved by LCM from FFPE tissues. This method is ideally suited for the analysis of relatively rare cell types within a tissue, and should improve on our ability to perform differential diagnosis of pathologies as compared to conventional LCM. Microsc. Res. Tech., 2009. © 2009 Wiley‐Liss, Inc.
Revitalization of a diastemal tooth primordium in Spry2 null mice results from increased proliferation and decreased apoptosis
Authors: Peterkova Renata , Churava Svatava , Lesot Herve , Rothova Michaela , Prochazka Jan , Peterka Miroslav , Klein Ophir D.
Published: May 2009

An understanding of the factors that promote or inhibit tooth development is essential for designing biological tooth replacements. The embryonic mouse dentition provides an ideal system for studying such factors because it consists of two types of tooth primordia. One type of primordium will go on to form a functional tooth, whereas the other initiates development but arrests at or before the bud stage. This developmental arrest contributes to the formation of the toothless mouse diastema. It is accompanied by the apoptosis of the rudimentary diastemal buds, which presumably results from the insufficient activity of anti‐apoptotic signals such as fibroblast growth factors (FGFs). We have previously shown that the arrest of a rudimentary tooth bud can be rescued by inactivating Spry2, an antagonist of FGF signaling. Here, we studied the role of the epithelial cell death and proliferation in this process by comparing the development of a rudimentary diastemal tooth bud (R2) and the first molar in the mandibles of Spry2−/− and wild‐type (WT) embryos using histological sections, image analysis and 3D reconstructions. In the WT R2 at embryonic day 13.5, significantly increased apoptosis and decreased proliferation were found compared with the first molar. In contrast, increased levels of FGF signaling in Spry2−/− embryos led to significantly decreased apoptosis and increased proliferation in the R2 bud. Consequently, the R2 was involved in the formation of a supernumerary tooth primordium. Studies of the revitalization of rudimentary tooth primordia in mutant mice can help to lay the foundation for tooth regeneration by enhancing our knowledge of mechanisms that regulate tooth formation. J. Exp. Zool. (Mol. Dev. Evol.) 312B:292–308, 2009. © 2009 Wiley‐Liss, Inc.
In vivo secretion of the mouse immunoglobulin G Fc fragment from rat submandibular glands
Authors: Racz Gabor Z. , Perez‐Riveros Paola , Adriaansen Janik , Zheng Changyu , Baum Bruce J.
Published: May 2009

Background

Salivary glands have been proposed as target organs for gene therapy. They secrete endogenous, as well as transgenic proteins, in a polarized manner. Transgene‐encoded regulated pathway proteins primarily follow the regulated pathway in rat salivary glands and are secreted into saliva in an exocrine manner. Conversely, constitutive pathway proteins generally are secreted more basolaterally and thus follow the endocrine route. In the present study, we studied in vivo the sorting of the mouse immunoglobulin G2b Fc fragment, which is physiologically secreted via the constitutive pathway.

Methods

Adenoviral vectors encoding the Fc fragment and human growth hormone were delivered into rat and mouse submandibular glands in vivo to compare their serum‐to‐saliva distribution. We also compared the intracellular localization of the Fc fragment and growth hormone by confocal microscopy.

Results

We found that the Fc fragment was secreted almost entirely into the bloodstream from rat and mouse submandibular glands via a constitutive or constitutive‐like pathway. This sorting behaviour is clearly different from that of transgenic human growth hormone, which is secreted in a regulated pathway, both in neuroendocrine cells and as a transgenic protein from salivary gland cells. We also found that simultaneously expressed human growth hormone and the mouse Fc fragment do not appear to influence each other's sorting behaviour. The Fc fragment showed a primarily basal localization, whereas growth hormone showed an apical localization, in rat submandibular gland acinar cells.

Conclusions

The results obtained in the present study indicate that the mouse Fc fragment is a useful model protein for examining the basolateral versus apical secretory pathways employed by transgenic secretory proteins in salivary glands. Copyright © 2009 John Wiley & Sons, Ltd.
Effects of nitric oxide and nitrogen on seedling emergence, ion accumulation, and seedling growth under salinity in the euhalophyte Suaeda salsa
Authors: Song Jie , Shi Gongwei , Xing Song , Chen Min , Wang Baoshan
Published: May 2009

Recently nitric oxide (NO) has emerged as a key signal molecule in plants. However, little is known about the role of NO in the salt tolerance of halophytes. Effects of the NO donors sodium nitroprusside (SNP) and nitrate (NO$ _3^- $) on growth and ion accumulation in the euhalophyte Suaeda salsa under salinity were investigated in the present study. The results showed that higher SNP supply increased seedling emergence, but SNP had no effect on shoot growth and the concentrations of Na+, K+, Cl–, and NO$ _3^- $. Higher NO$ _3^- $ had no effect on seedling emergence of the species. Shoot Cl– decreased, but NO3– increased markedly, with a higher NO$ _3^- $ supply. The decrease in the estimated contribution of Cl– to the osmotic potential was compensated for by an increase in that of NO$ _3^- $. It appears that NO$ _3^- $ plays an important osmotic role in S. salsa under high salinity with a higher NO$ _3^- $ supply, and this trait may increase salt tolerance of the species under high salinity.
Pore‐size distribution and particle arrangement in fragipan and nonfragipan horizons
Authors: Falsone Gloria , Bonifacio Eleonora
Published: May 2009

Fragipan is a widely distributed subsoil horizon that induces severe limitations to plant growth and land use, mainly because of its high bulk density. In this work, we evaluated the pore‐size distribution through the analysis of the cumulative curve of intruded mercury volume in some soils with fragipan horizons. This approach provides information also about the arrangement of particles, thus we compared the results obtained for fragipan and nonfragipan horizons to relate porosity and particle arrangement with the specific physical properties of fragipans. The total volume of intruded mercury did not allow to discriminate between fragipan and nonfragipan horizons. However, from the variation of the pore volume as a function of the radius, two modal classes of pores were found, coarse and fine, respectively. The fine‐pore class arose from the arrangement of clay particles, and its volume was correlated to clay contents (r = 0.787) and to clay packing density (r = –0.621). The clay fractions in fragipans were less densely packed than in the other B horizons, even if they had similar clay contents. The coarser‐pore modal class is known to arise from the interactions between clay, silt, and sand particles, and its volume was different among horizons. Fragipan had a low volume of this modal pore class. In addition, a packing density for the coarser phase of 0.74, corresponding to a rhombohedral packing, was found only in fragipans. Thus, the low permeability and high bulk density of fragipans are linked to specific arrangements of the particles: an open packing of the clay phase is associated to an extremely dense packing of silt and sand. This combination is not present in any other soil horizon.
Arsenic accumulation by rice grown in soil treated with roxarsone
Authors: Liu Chen‐Wuing , Lin Ching‐Chieh , Jang Cheng‐Shin , Sheu Guey‐Rong , Tsui Lo
Published: May 2009

Poultry litter is widely used as a fertilizer for lowland rice in Taiwan and China. However, the organic‐arsenic compound roxarsone (additive of poultry feed) in poultry litter can be absorbed by the plants and the resulting arsenic (As) contamination may pose a serious threat to human health. This study used various amounts of poultry litter contaminated with roxarsone in pot experiments to evaluate the effect of roxarsone on rice agronomic parameters and the bioaccumulation of total and inorganic As in rice‐plant tissues. Rice‐grain yield decreased significantly with increasing As content of the soil, and the critical threshold that killed rice was 200 mg roxarsone (kg soil)–1. The As concentrations in root, straw, leaf, husk, and grain increased with increasing soil As (p< 1%). At 100 mg roxarsone per kg of soil, the As concentration in the rice grain exceeded the statutory permissible limit of 1.0 mg As (kg dry weight)–1and at 25 mg roxarsone (kg soil)–1, the inorganic As concentrations in grains exceeded the statutory limit of 0.15 mg of inorganic As kg–1in China. For all treatments, the As concentrations in various plant tissues at maturity follow the order: root > stem > leaf > husk > grain. Arsenite was the predominant species in root, straw, and grain, while arsenate was the predominant species in leaf and husk. No significant difference existed between the amounts of arsenite and arsenate when various amounts of poultry litter were applied. This result illustrates that large amounts of added roxarsone are not only toxic to rice but also accumulate in grains in the inorganic As forms, potentially posing a threat to human healthviathe food chain.
An efficient combination of supercritical fluid extraction and high‐speed counter‐current chromatography to extract and purify homoisoflavonoids from Ophiopogon japonicus (Thunb.) Ker‐Gawler
Authors: Ma Chengjun , Li Gang , Zhang Juan , Zheng Qiusheng , Fan Xiao , Wang Zhenhua
Published: May 2009

Supercritical fluid extraction (SFE) was used to extract homoisoflavonoids from Ophiopogon japonicus (Thunb.) Ker‐Gawler. The optimization of parameters was carried out using an orthogonal test L9 (3)4 including pressure, temperature, dynamic extraction time and the amount of modifier. The process was then scaled up by 100 times with a preparative SFE system under the optimized conditions of 25 MPa, 55°C, 4.0 h and 25% methanol as a modifier. Then crude extracts were separated and purified by high‐speed counter‐current chromatography (HSCCC) with a two‐phase solvent system composed of n‐hexane/ethyl acetate/methanol/ACN/water (1.8:1.0:1.0:1.2:1.0 v/v). There three homoisoflavonoidal compounds including methylophiopogonanone A 6‐aldehydo‐isoophiopogonone A, and 6‐formyl‐isoophiopogonanone A, were successfully isolated and purified in one step. The collected fractions were analyzed by HPLC. In each operation, 140 mg crude extracts was separated and yielded 15.3 mg of methylophiopogonanone A (96.9% purity), 4.1 mg of 6‐aldehydo‐isoophiopogonone A (98.3% purity) and 13.5 mg of 6‐formyl‐isoophiopogonanone A (97.3% purity) respectively. The chemical structure of the three homoisoflavonoids are identified by means of ESI‐MS and NMR analysis.
The self‐presentation and self‐development of serious wildlife tourists
Authors: Curtin Susanna
Published: May 2009

The notion of self‐concept and choice of tourism products has yet to be applied to wildlife tourism. The question of how consumers perceive themselves is considered fundamental to understanding purchase and consumption behaviours. This ethnographic study is based upon ‘serious’ wildlife tourism participants, for whom studying fauna and flora is the primary motivation for travel. The findings demonstrate how the ‘culture’ of ‘serious’ wildlife tourism is made up of individuals who differentiate themselves from other tourists in terms of dress, behaviour, development of skills, equipment and intellectual capital, illustrated by their desire to scope, identify and photograph wildlife. Copyright © 2009 John Wiley & Sons, Ltd.
Changes in soil nitrogen availability due to stand development and management practices on semi‐arid sandy lands, in northern China
Authors: Chen F. S. , Zeng D. H. , Fahey T. J.
Published: May 2009

Soil nitrogen (N) availability is one of the limiting factors for plant growth on sandy lands. Little is known about impacts of afforestation on soil N availability and its components in southeastern Keerqin sandy lands, China. In this study, we measured N transformation under sandy Mongolian pine (Pinus sylvestris var. mongolica Litv.) plantations of different ages (grassland, young, middle‐aged, close‐to‐mature) and management practices (non‐grazing and free‐grazing) during the growing seasons using the ion exchange resin bag method. Results showed that, for all plots and growing season, soil NH‐N, NO‐N, mineral N, and relative nitrification index, varied from 0·18 to 1·54, 0·96 to 22·05, 1·23 to 23·58 µg d−1 g−1 dry resin, and 0·76 to 0·97, respectively, and NO‐N dominated the available N amount due to intense nitrification in these ecosystems. In general, the four indices significantly increased in the oldest plantation, with corresponding values in non‐grazing sites lower than those in free‐grazing sites (p < 0·05). Our studies indicated that it is a slow, extended process to achieve improvement in soil quality after afforestation of Mongolian pine in the study area. Copyright © 2009 John Wiley & Sons, Ltd.
The association between mutations in the lysosomal protein glucocerebrosidase and parkinsonism
Authors: DePaolo John , Goker‐Alpan Ozlem , Samaddar Ted , Lopez Grisel , Sidransky Ellen
Published: May 2009

A body of work has emerged over the past decade demonstrating a relationship between mutations in glucocerebrosidase gene (GBA), the gene implicated in Gaucher disease (GD), and the development of parkinsonism. Several different lines of research support this relationship. First, patients with GD who are homozygous for mutations in GBA have a higher than expected propensity to develop Parkinson's disease (PD). Furthermore, carriers of GBA mutations, particularly family members of patients with GD, have displayed an increased rate of parkinsonism. Subsequently, investigators from centers around the world screened cohorts of patients with parkinsonism for GBA mutations and found that overall, subjects with PD, as well as other Lewy body disorders, have at least a fivefold increase in the number of carriers of GBA mutations as compared to age‐matched controls. In addition, neuropathologic studies of subjects with parkinsonism carrying GBA mutations demonstrate Lewy bodies, depletion of neurons of the substantia nigra, and involvement of hippocampal layers CA2–4. Although the basis for this association has yet to be elucidated, evidence continues to support the role of GBA as a PD risk factor across different centers, synucleinopathies, and ethnicities. Further studies of the association between GD and parkinsonism will stimulate new insights into the pathophysiology of the two disorders and will prove crucial for both genetic counseling of patients and family members and the design of relevant therapeutic strategies for specific patients with parkinsonism. © 2009 Movement Disorder Society
Clinical and physiological assessments for elucidating falls risk in Parkinson's disease
Authors: Latt Mark D. , Lord Stephen R. , Morris John G.L. , Fung Victor S.C.
Published: May 2009

The study aims were to devise (1) a fall risk screen for people with PD using routine clinical measures and (2) an explanatory (physiological) fall risk assessment for guiding fall prevention interventions. One hundred thirteen people with PD (age 66 ± 95% CI 1.6 years) underwent clinical assessments and quantitative tests of sway, gait, strength, reaction time, and lower limb sensation. Participants were then followed up for 12 months to determine fall incidence. In the follow‐up year, 51 participants (45%) fell one or more times whereas 62 participants (55%) did not fall. Multivariate analyses of routine clinical measures revealed that a fall in the past year, abnormal axial posture, cognitive impairment, and freezing of gait were independent risk factors for falls and predicted 38/51 fallers (75%) and 45/62 non‐fallers (73%). A multivariate model combining clinical and physiological measures that elucidate the pathophysiology of falls identified abnormal posture, freezing of gait, frontal impairment, poor leaning balance, and leg weakness as independent risk factors. This model correctly classified 39/51 fallers (77%) and 51/62 non‐fallers (82%). Patients with PD at risk of falls can be identified accurately with routine clinical assessments and quantitative physiological tests. Many of the risk factors identified are amenable to targeted intervention. © 2009 Movement Disorder Society
Alzheimer's disease and corticobasal degeneration presenting as corticobasal syndrome
Authors: Hu William T. , Rippon Gregory W. , Boeve Bradley F. , Knopman David S. , Petersen Ronald C. , Parisi Joseph E. , Josephs Keith A.
Published: May 2009

The aim of this article is to compare patients with Alzheimer's disease (AD) pathology and corticobasal degeneration pathology (CBD) presenting as corticobasal syndrome (CBS). Clinicopathologic series was used. Five patients with AD and 11 patients with CBD were clinically diagnosed with CBS. Patients with AD pathology had an earlier age of onset than patients with CBD pathology (58 vs. 68 years, P = 0.004), but the two groups had similar disease duration and core features of CBS. Tremors were only present in CBD cases (73%, P = 0.026), but myoclonus was more common in AD than CBD (80 vs. 18%, P = 0.036). Neuropsychological testing showed similar degrees of memory impairment and attentional deficits. 99mTc‐HMPAO SPECT imaging demonstrated parietal hypoperfusion in AD patients and frontotemporal hypoperfusion in CBD patients. AD patients with clinical CBS have similar characteristics to CBD patients. Functional brain imaging may have greater utility than the clinical and neuropsychological features in differentiating AD presenting as CBS from CBD. © 2009 Movement Disorder Society
Evolution of stereotypies in adolescents and women with Rett syndrome
Authors: Vignoli Aglaia , La Briola Francesca , Canevini Maria Paola
Published: May 2009

Stereotypies in Rett syndrome (RTT) are a diagnostic hallmark present in all stages of the disease, but descriptions of movement disorders in adults are very scant. Among 30 patients with RTT followed‐up at San Paolo Hospital in Milan, we selected those aged ≥14 years and studied 12 patients (mean age 18. 6 years, range: 14–31) with MECP2 mutations. Mean age at stereotypies onset was 19.4 months; stereotypies at onset tend to be maintained during evolution, while new stereotyped movements can be detected in the follow‐up. All patients still present stereotypies involving separated or joined hands: most frequently mouthing, pill rolling, and twisting. We underline that stereotyped movements persist in older patients and can be useful to suspect RTT diagnosis in adult age in otherwise unclassified patients. © 2009 Movement Disorder Society
Bradykinesia, muscle weakness and reduced muscle power in Parkinson's disease
Authors: Allen Natalie E. , Canning Colleen G. , Sherrington Catherine , Fung Victor S.C.
Published: May 2009

Muscle power (force × velocity) could clarify the relationship between weakness and bradykinesia in Parkinson's disease (PD). The aims of this study were to determine if patients with PD were weaker and/or less powerful in their leg extensor muscles than a neurologically normal control group and to determine the relative contributions of force and movement velocity/bradykinesia to muscle power in PD. Forty patients with PD and 40 controls were assessed. Strength in Newtons (N) was measured as the heaviest load the participant could lift. Power in Watts (W) was measured by having the participant perform lifts as fast as possible. The PD group were 172 N weaker (95% CI 28–315) and 124 W less powerful at peak power (95% CI 32–216) than controls. However, velocity at maximal power was only reduced compared with controls when lifting light to medium loads. When lifting heavy loads bradykinesia was no longer apparent in the PD group. These results suggest that reduced muscle power in PD at lighter loads arises from weakness and bradykinesia combined, but at heavier loads arises only from weakness. The absence of bradykinesia in the PD group when lifting heavy loads warrants further investigation. © 2009 Movement Disorder Society