bluuebunny/crossref_metadata_2025 huggingface dataset:
Functional and molecular mapping of uncoupling between vascular permeability and loss of vascular maturation in ovarian carcinoma xenografts: The role of stroma cells in tumor angiogenesis
Authors: Gilad Assaf A. , Israely Tomer , Dafni Hagit , Meir Gila , Cohen Batya , Neeman Michal
Published: May 2005

Maintaining homogeneous perfusion in tissues undergoing remodeling and vascular expansion requires tight orchestration of the signals leading to endothelial sprouting and subsequent recruitment of perivascular contractile cells and vascular maturation. This regulation, however, is frequently disrupted in tumors. We previously demonstrated the role of tumor‐associated myofibroblasts in vascularization and exit from dormancy of human ovarian carcinoma xenografts in nude mice. The aim of this work was to determine the contribution of stroma‐ and tumor cell‐derived angiogenic growth factors to the heterogeneity of vascular permeability and maturation in MLS human ovarian carcinoma tumors. We show by RT‐PCR and by in situ hybridization that VEGF was expressed by the tumor cells, while angiopoietin‐1 and ‐2 were expressed only by the infiltrating host stroma cells. Vascular maturation was detected in vivo by vasoreactivity to hypercapnia, measured by BOLD contrast MRI and validated by immunostaining of histologic sections to α‐smooth muscle actin. Vascular permeability was measured in vivo by dynamic contrast‐enhanced MRI using albumin‐based contrast material and validated in histologic sections by fluorescent staining of the biotinylated contrast material. MRI as well as histologic correlation maps between vascular maturation and vascular permeability revealed a wide range of vascular phenotypes, in which the distribution of vascular maturation and vasoreactivity did not overlap spatially with reduced permeability. The large heterogeneity in the degree of vascular maturation and permeability is consistent with the differential expression pattern of VEGF and angiopoietins during tumor angiogenesis. © 2005 Wiley‐Liss, Inc.
Oxidative addition of the ethane CC bond to Pd. An ab initio benchmark and DFT validation study
Authors: De Jong G. Theodoor , Geerke Daan P. , Diefenbach Axel , Solà Miquel , Bickelhaupt F. Matthias
Published: May 2005

We have computed a state‐of‐the‐art benchmark potential energy surface (PES) for the archetypal oxidative addition of the ethane CC bond to the palladium atom and have used this to evaluate the performance of 24 popular density functionals, covering LDA, GGA, meta‐GGA, and hybrid density functionals, for describing this reaction. The ab initio benchmark is obtained by exploring the PES using a hierarchical series of ab initio methods [HF, MP2, CCSD, CCSD(T)] in combination with a hierarchical series of five Gaussian‐type basis sets, up to g polarization. Relativistic effects are taken into account either through a relativistic effective core potential for palladium or through a full four‐component all‐electron approach. Our best estimate of kinetic and thermodynamic parameters is −10.8 (−11.3) kcal/mol for the formation of the reactant complex, 19.4 (17.1) kcal/mol for the activation energy relative to the separate reactants, and −4.5 (−6.8) kcal/mol for the reaction energy (zero‐point vibrational energy‐corrected values in parentheses). Our work highlights the importance of sufficient higher angular momentum polarization functions for correctly describing metal‐d‐electron correlation. Best overall agreement with our ab initio benchmark is obtained by functionals from all three categories, GGA, meta‐GGA, and hybrid DFT, with mean absolute errors of 1.5 to 2.5 kcal/mol and errors in activation energies ranging from −0.2 to −3.2 kcal/mol. Interestingly, the well‐known BLYP functional compares very reasonably with a slight underestimation of the overall barrier by −0.9 kcal/mol. For comparison, with B3LYP we arrive at an overestimation of the overall barrier by 5.8 kcal/mol. On the other hand, B3LYP performs excellently for the central barrier (i.e., relative to the reactant complex) which it underestimates by only −0.1 kcal/mol. © 2005 Wiley Periodicals, Inc. J Comput Chem 26: 1006–1020, 2005
DFT study and Monte Carlo simulation on proton transfers of 2‐amino‐2‐oxazoline, 2‐amino‐2‐thiazoline, and 2‐amino‐2‐imidazoline in the gas phase and in water
Authors: Xue Ying , Kim Chan Kyung , Guo Yong , Xie Dai Qian , Yan Guo Sen
Published: May 2005

Density functional theory (DFT) and Monte Carlo (MC) simulation with free energy perturbation (FEP) techniques have been used to study the tautomeric proton transfer reaction of 2‐amino‐2‐oxazoline, 2‐amino‐2‐thiazoline, and 2‐amino‐2‐imidazoline in the gas phase and in water. Two reaction pathways were considered: the direct and water‐assisted transfers. The optimized structures and thermodynamic properties of stationary points for the title reaction system in the gas phase were calculated at the B3LYP/6‐311+G(d, p) level of theory. The potential energy profiles along the minimum energy path in the gas phase and in water were obtained. The study of the solvent effect of water on the proton transfer of 2‐amino‐2‐oxozoline, 2‐amino‐2‐thiazoline, and 2‐amino‐2‐imidazoline indicates that water as a solvent is favorable for the water‐assisted process and slows down the rate of the direct transfer pathway. © 2005 Wiley Periodicals, Inc. J Comput Chem 26: 994–1005, 2005
Clinical psychologists across the years: The division of clinical psychology from 1960 to 2003
Authors: Norcross John C. , Karpiak Christie P. , Santoro Shannon O.
Published: May 2005

For more than 40 years researchers have studied the members of the American Psychological Association's (APA) Division of Clinical Psychology to obtain information about their demographic characteristics, educational experiences, theoretical orientations, employment settings, professional activities, publication histories, and career satisfactions. We summarize the results from the most recent study (N = 694, 46% return rate) in the historical context of the previous findings, dating back to 1960. We provide both contemporary and historical portraits of American clinical psychology. Among the most notable trends are a steady increase in female psychologists, a decline in psychological assessment in general and projective testing in particular, the modal eclectic/integrative orientation being rivaled by the cognitive orientation, a pattern of high career satisfaction, and continued enthusiasm for the Boulder model. © 2005 Wiley Periodicals, Inc. J Clin Psychol 61: 1467–1483, 2005.
Disruption of protein–protein interactions: Towards new targets for chemotherapy
Authors: Loregian Arianna , Palù Giorgio
Published: May 2005

Protein–protein interactions play a key role in various mechanisms of cellular growth and differentiation, and in the replication of pathogen organisms in host cells. Thus, inhibition of these interactions is a promising novel approach for rational drug design against a wide number of cellular and microbial targets. In the past few years, attempts to inhibit protein–protein interactions using antibodies, peptides, and synthetic or natural small molecules have met with varying degrees of success, and these will be the focus of this review. © 2005 Wiley‐Liss, Inc.
Loss of E‐cadherin mediated cell–cell adhesion as an early trigger of apoptosis induced by photodynamic treatment
Authors: Galaz Sergio , Espada Jesús , Stockert Juan C. , Pacheco María , Sanz‐Rodríguez Francisco , Arranz Rocío , Rello Santiago , Cañete Magdalena , Villanueva Angeles , Esteller Manel , Juarranz Angeles
Published: May 2005

Photodynamic treatment with different photosensitizers (PSs) can result in the specific induction of apoptosis in many cell types. It is commonly accepted that this apoptotic response depends on the mitochondrial accumulation of the PS. Accumulation in other cellular organelles, such as lysosomes or the Golgi complex, and subsequent photodamage resulting in an apoptotic process has been also described. However, the role played by cell adhesion in apoptosis induced in epithelial cells after photodynamic treatment is not well characterized. Here, we have used a murine keratinocyte line, showing a strong dependence on E‐cadherin for cell–cell adhesion and survival, to analyze the relevance of this adhesion complex in the context of zinc(II)‐phthalocyanine (ZnPc) photodynamic treatment. We report that under apoptotic conditions, ZnPc phototreatment induces a rapid disorganization of the E‐cadherin mediated cell–cell adhesion, which largely preceded both the detachment of cells from the substrate, via β‐1 integrins and the induction of apoptotic mitochondrial markers. Therefore, the alteration in E‐cadherin, α‐ and β‐catenins adhesion proteins preceded the release of cytochrome c (cyt c) from mitochondria to the cytosol and the activation of caspase 3. In addition, blocking E‐cadherin function with a specific antibody (Decma‐1) induced apoptosis in this cell system. These results strongly suggest that the E‐cadherin adhesion complex could be the primary target of ZnPc phototreatment, and that loss of E‐cadherin mediated cell adhesion after early photodamage triggers an apoptotic response. © 2005 Wiley‐Liss, Inc.
Mdm2 and HIF‐1α interaction in tumor cells during hypoxia
Authors: Nieminen Anna‐Liisa , Qanungo Suparna , Schneider Elizabeth A. , Jiang Bing‐Hua , Agani Faton H.
Published: May 2005

The interaction between HIF‐1α, Mdm2, and p53 proteins during hypoxia has received recent attention. Here, we investigated the consequences of interaction between HIF‐1α and Mdm2 under hypoxic conditions. Endogenous HIF‐1α and Mdm2 proteins were co‐immunoprecipitated from lysates of hypoxic HCT116 p53WT and p53−/− cells, suggesting that association of these two proteins is a p53‐independent event. The cellular Mdm2 protein content was not significantly altered in hypoxic tumor cells. Overexpression of Mdm2 resulted in an increase in HIF‐1α protein content in hypoxic cells and increased hypoxia‐induced vascular endothelial growth factor (VEGF) transcriptional activation. These results point toward a novel and p53‐independent function of Mdm2 to promote tumor cell adaptations to hypoxia by interacting with and promoting HIF‐1 activation. © 2005 Wiley‐Liss, Inc.
Performance of H‐ZSM‐5‐supported bimetallic catalysts for the combustion of polluting volatile organic compounds in air
Authors: Abdullah Ahmad Zuhairi , Abu Bakar Mohamad Zailani , Bhatia Subhash
Published: May 2005

The performance of H‐ZSM‐5‐supported bimetallic catalysts with chromium as the base metal in the combustion of ethyl acetate and benzene is reported. A reactor operated from 100 to 500 °C at a gas hourly space velocity (GHSV) of 32 000 h−1 was used for study of the activity. A combination of 1.0 wt% chromium and 0.5 wt% copper yielded a catalyst (Cr1.0Cu0.5/Z) with improved conversion and carbon dioxide yield. Cr2O3 (Cr3+) and CuO (Cu2+) were the predominant metal species in the catalyst. In agreement with the Mars–van Krevelen model, improved reducibility of Cr3+ in the presence of Cu2+ led to an improvement in activity. The copper content in Cr1.0Cu0.5/Z also favored the formation of deep combustion products. Condensation and subsequent growth of coke precursors in the catalyst pores led to the formation of a softer and less aromatic coke fraction while dehydrogenation activity on acid sites formed a harder and more aromatic coke fraction. The use of Cr1.0Cu0.5/Z favored the formation of lower molecular weight intermediates, leading to reduction in formation of softer coke. Copyright © 2005 Society of Chemical Industry
Synthesis and characterization of a synthetic heme‐based oxygen carrier
Authors: Al‐Kassim Loola , Wang Z , Szabo Arthur G , Taylor Keith E , Mutus Bulent
Published: May 2005

We have prepared a novel oxygen carrier composed of Starburst dendrimer 3 hematoporphyrin iron(III) di‐1‐(3‐aminopropyl)imidazole di‐succinic acid (SDHAPI). This compound has a molecular weight of ∼12.7 kD. O2 and CO binding to SDHAPI was demonstrated by UV‐vis spectroscopy. Oxygenated SDHAPI had an estimated auto‐oxidation half‐life of 24 min. The oxygen stoichiometry (∼6 mol of O2 mol−1 SDHAPI) and the kobs(on+off) of 0.084 s−1 and kobs(off) of 0.009 s−1 were estimated with an oxygen electrode. Copyright © 2005 Society of Chemical Industry
Application of artificial neural networks to the analysis of two‐dimensional fluorescence spectra in recombinant E coli fermentation processes
Authors: Lee Kum‐Il , Yim Yong‐Sik , Chung Sang‐Wook , Wei Jiaqiu , Rhee Jong Il
Published: May 2005

A two‐dimensional (2D) spectrofluorometer was used to monitor various fermentation processes with recombinant E coli for the production of 5‐aminolevulinic acid (ALA). The whole fluorescence spectral data obtained during a process were analyzed using artificial neural networks, ie self‐organizing map (SOM) and feedforward backpropagation neural network (BPNN). The SOM‐based classification of the whole spectral data has made it possible to qualitatively associate some process parameters with the normalized weights and variances, and to select some useful combinations of excitation and emission wavelengths. Based on the classified fluorescence spectra a supervised BPNN algorithm was used to predict some of the process parameters. It was also shown that the BPNN models could elucidate some sections of the process's performance, eg forecasting the process's performance. Copyright © 2005 Society of Chemical Industry
Adsorption of selected environmentally important metals by poultry manure‐based granular activated carbons
Authors: Lima Isabel M , Marshall Wayne E
Published: May 2005

Water quality and public health impacts of mass produced poultry manure have prompted the need for viable conversion and reuse solutions. Conversion of poultry manure to value‐added granular activated carbons for environmental remediation can be such a solution. The objective of this study was to compare the effectiveness of poultry manure‐based carbons for the adsorption of selected metal ions (copper, cadmium, nickel, zinc) from solutions containing individual ions or the four metal ions present together. Adsorption properties for poultry manure‐based carbons were compared with those of two commercial carbons, PŪR RF and Minotaur, and carbons from three traditional precursors, coal, coconut and wood. Pelletized samples were pyrolyzed at 700 °C for 1 h followed by 800 °C steam activation at 1, 3 and 5 mL min−1 water flow rate, for 30 and 45 min, under nitrogen. The carbon's ability to adsorb the metals was influenced by the activation strategy, increasing for longer activation times and higher water flow rates, in the presence of a single metal solution. Saturation conditions were achieved at 20 mM and, at 5 mM, almost all metal ion in solution was adsorbed, except for nickel. Carbons showed similar affinity towards copper, cadmium and zinc, individually. However, when in competition, at 5 mM of each metal, manure‐based carbons showed a preference for Cu2+ followed by Zn2+ and Cd2+. Poultry manure‐based carbons outperformed all reference carbons and PŪR RF, and, except for Ni2+, Minotaur was also outperformed. Experiments showed that the highest removal rate in a competition situation was obtained for broiler‐cake carbon activated for 45 min at 3 mL min−1, with 93.1%, 50.9% and 85.2% for copper, cadmium and zinc ions, respectively. Published in 2005 for SCI by John Wiley & Sons, Ltd.
Oxygen transfer effects in β‐lactamase fermentation by Bacillus licheniformis in a glucose‐based defined medium
Authors: Çalık Pınar , Arifoǧlu Müge , Çalık Güzide
Published: May 2005

The effects of oxygen transfer on the β‐lactamase production by Bacillus licheniformis were investigated in a glucose‐based defined medium. The experiments were conducted in 3.0 dm3 batch bioreactor systems at three different air inlet (QO/VR = 0.2, 0.5 and 1.0 vvm) and agitation rates (N = 250, 500 and 750 min−1). During the fermentation, the concentrations of the cell, glucose, by‐products, ie organic and amino acids, oxygen transfer coefficients (KLa), yield coefficients, specific rates and oxygen uptake rates (OUR) were determined, in addition to β‐lactamase activities. The highest β‐lactamase activity was obtained at QO/VR = 0.5 vvm and N = 500 min−1 and at QO/VR = 0.2 vvm and N = 500 min−1 conditions, as ca A = 90 U cm−3. The highest cell concentration was obtained as CX = 0.67 kg m−3 at QO/VR = 0.5 vvm and N = 750 min−1 and at QO/VR = 0.2 vvm and N = 750 min−1 conditions. The values of KLa increased with increasing agitation and aeration rates and varied between 0.007 and 0.044 s−1, and the OUR varied between 0.4 and 1.6 mol m−3 s−1. With increasing QO/VR and/or N, the Damköhler number (ie the oxygen transfer limitation) decreased owing to the increase in mass transfer coefficients (KLa). The highest instantaneous yield of cell on substrate (YX/S) and yield of cell on oxygen (YX/O) values were respectively obtained at 0.5 vvm and 500 min−1 conditions at t = 2 h as YX/S = 0.72 kg kg−1 and YX/O = 1.49 kg kg−1. The highest instantaneous yield of substrate on oxygen (YS/O) was obtained at 0.5 vvm and 750 min−1 conditions at t = 20 h as YS/O = 8.07 kg kg−1. Copyright © 2005 Society of Chemical Industry
Catalytic hydrothermal treatment of pine wood biomass: effect of RbOH and CsOH on product distribution
Authors: Karagöz Selhan , Bhaskar Thallada , Muto Akinori , Sakata Yusaku
Published: May 2005

Low‐temperature hydrothermal treatment of pine wood biomass was performed in the presence of RbOH and CsOH catalysts (280 °C for 15 min). The effect of the catalysts on the distribution of products and the volatility distribution of oxygenated hydrocarbons was studied in detail. Oxygenated hydrocarbons were extracted from the liquid and solid portions and analysed individually by gas chromatography/mass spectrometry. Catalytic (RbOH and CsOH) hydrothermal treatment of wood biomass produced mainly phenolic compounds and benzenediol derivatives. The use of RbOH and CsOH catalysts hindered the formation of char and favoured the formation of oil products, as observed previously for various other base catalysts. The volatility distribution of hydrocarbons (ether extract) was characterised by carbon‐normal paraffin (C‐NP) gram and it was found that the oxygenated hydrocarbons from all runs, including thermal, were distributed in the boiling point region of n‐C6 to n‐C17. Copyright © 2005 Society of Chemical Industry
Simultaneous treatment of semiconductor wastewater and distillery slops by mixing and precipitation/coagulation
Authors: Navarro Ronald R , Navarro Rosvic C , Alfafara Catalino G , Demafelis Rex B , Tatsumi Kenji
Published: May 2005

Organic matter (chemical oxygen demand, COD) removal with decolorization in waste distillery slops and copper removal in a semiconductor industry wastewater were achieved in a single step mixing and precipitation/coagulation treatment system. The process utilized the complementary properties of the positively charged copper ions in semiconductor wastewater and net negative charge of melanoidin (organic chromophoric pollutant) in distillery slops to mutually neutralize each other. Copper ions served as coagulant for slops and melanoidin served as precipitant for copper. The volumetric ratio of the wastewater and pH were optimized to attain maximum removal of organic matter and copper. The optimum volumetric ratio for the evaluated semiconductor wastewater to distillery slops was found to be 2–3 with an equilibrium pH of around pH 6. At optimum conditions, average removals of COD and copper were 86% and 92%, respectively, in an actual and undiluted system. Decolorization efficiency using the diluted distillery slops was 89%. The process can be considered an effective pretreatment procedure for simultaneous gross removal of copper and color/COD, particularly in highly concentrated waste streams. Copyright © 2005 Society of Chemical Industry
Granulation of a mixture of suspended anaerobic and aerobic cultures under alternating anaerobic/microaerobic/aerobic conditions: a preliminary study
Authors: Ergüder Tuba H , Demirer Göksel N
Published: May 2005

This paper focuses on a new granulation study area where a mixture of suspended anaerobic and aerobic cultures was used as seed. Mixed suspended anaerobic and aerobic cultures exposed to alternating anaerobic/microaerobic/aerobic conditions developed granulates up to 2.5 mm in size. The granules developed displayed both oxygen uptake (3.83–8.75 mg dissolved oxygen g−1 volatile suspended solids h−1) and methanogenic activities (7.5 cm3 CH4 d−1) comparable to those of aerobic and anaerobic control cultures, respectively. Oxygen doses of 60, 100 and 120% of the total chemical oxygen demand added (23, 38 and 45 cm3) and ethanol (among ethanol, glucose, acetic acid, and glucose + acetic acid) resulted in the largest and the most stable granules. Copyright © 2005 Society of Chemical Industry
Role of unburnt carbon in adsorption of dyes on fly ash
Authors: Wang Shaobin , Boyjoo Yashveersingh , Choueib Ali , Ng Esther , Wu Hongwei , Zhu Zhonghua
Published: May 2005

Various fly ash samples with different unburnt carbon contents were collected, characterised and tested for adsorption of basic dyes, Methylene Blue and Crystal Violet, in aqueous solution. It was found that unburnt carbon plays a major role in dye adsorption. The mineral matter of fly ash has little adsorption capacity and most of the adsorption capacity of fly ash can be attributed to the unburnt carbon. The fly ash with higher unburnt carbon content will have higher adsorption capacity. For the carbon‐free fly ash, adsorption capacities for Methylene Blue and Crystal Violet are only 2 × 10−6 mol g−1 and 1.0 × 10−6 mol g−1, respectively, while the adsorption capacities for Methylene Blue and Crystal Violet on carbon‐enriched fly ash are 1.2 × 10−4 mol g−1 and 1.0 × 10−4 mol g−1, respectively. A two‐site Langmuir adsorption model best describes the adsorption isotherm. Copyright © 2005 Society of Chemical Industry
Pretreatment of Chromobacterium viscosum lipase with acetone increases its activity in sodium bis‐(2‐ethylhexyl) sulfosuccinate (AOT) reverse micelles
Authors: Talukder MMR , Zaman MM , Hayashi Y , Wu JC , Kawanishi T
Published: May 2005

The activity of Chromobacterium viscosum lipase for hydrolysis of olive oil in sodium bis‐(2‐ethylhexyl) sulfosuccinate (AOT) reverse micelles was increased by pretreatment with acetone. In contrast to the untreated lipase, no sharp fall in the activity of the treated lipase at higher W0 (water to AOT molar ratio) values was observed. The fluorescence emission intensity of the treated lipase in reverse micelles was higher than that of the untreated lipase but the maximal emission wavelength (λmax) was the same for both lipases. A kinetic model that considers the free substrate in equilibrium with the substrate adsorbed on the micellar surface was successfully used to better understand the activity enhancement. The Michaelis constant (Km) and substrate adsorption equilibrium constant (Kad) were reduced by lipase pretreatment with acetone whereas the maximum reaction rate (vmax) remained unaltered. Copyright © 2005 Society of Chemical Industry
Comparison of chemical wet scrubbers and biofiltration for control of volatile organic compounds using GC/MS techniques and kinetic analysis
Authors: Kastner James R , Das Keshav C
Published: May 2005

Increasing public concerns and EPA air regulations in non‐attainment zones necessitate the remediation of volatile organic compounds (VOCs) generated in the poultry‐rendering industry. Wet scrubbers using chlorine dioxide (ClO2) have low overall removal efficiencies due to lack of reactivity with aldehydes. Contrary to wet scrubbers, a biofilter system successfully treated the aldehyde fraction, based on GC/MS analysis of inlet and outlet streams. Total VOC removal efficiencies ranged from 40 to 100% for the biofilter, kinetic analysis indicated that the overall removal capacity approached 25 g m−3 h−1, and aldehyde removal efficiency was significantly higher compared with chemical wet scrubbers. Process temperatures monitored in critical unit operations upstream from the biofilter varied significantly during operation, rising as much as 30 °C within a few minutes. However, the outlet air temperature of a high intensity scrubber remained relatively constant at 40 °C, although the inlet air temperature fluctuated from 50 to 65 °C during monitoring. These data suggest a hybrid process combining a wet scrubber and biofilter in series could be used to improve overall VOC removal efficiencies and process stability. Copyright © 2005 Society of Chemical Industry
Effect of scale‐up on wall shear rates in circulating bubble columns
Authors: Al‐Masry Waheed A , Al‐Ahmed Malik I
Published: May 2005

Effective wall shear rates were investigated experimentally in an external loop circulating bubble column made from transparent acrylic resin. The riser (DR) and downcomer (DD) diameters were 0.19 m and 0.14 m, respectively. The column working volume (VR) was 170 dm3, with a scale‐up factor (AD/AR) of 0.54, and a dispersion height (HD) of 2.25 m. Polymer solutions of xanthan gum and carboxymethyl cellulose were used to simulate non‐Newtonian behavior of biological systems. Effective wall shear rates for the non‐Newtonian solutions were found by analogy with Newtonian glycerol solutions, employing downcomer liquid velocity as the measurable and comparable parameter. The experimental shear rate results were found to fit between those of the literature data. A new single correlation taking into account all the relevant data in the literature and the results of this work is proposed. The new correlation is an improvement over the other correlations because it includes two important design constants: scale‐up factor and dispersion height. Copyright © 2005 Society of Chemical Industry
A comprehensive study of optimal conditions for naked plasmid DNA transfer into skeletal muscle by electroporation
Authors: Wang Xue‐Dong , Tang Jian‐Guo , Xie Xiao‐lei , Yang Ji‐Chun , Li Shuai , Ji Jian‐Guo , Gu Jun
Published: May 2005

Efficient gene transfer is a key factor in gene therapy. Reducing the damage caused by gene transfer to muscle by electroporation is very important for its clinical application. Extensive investigation of optimal conditions for gene transfer by electroporation is required. The parameters used for electroporation, including plasmid concentration; injection volume; the plasmid dose of the injection; the concentration of saline media; the size of plasmid DNA; the age of the mice; the lag time between plasmid injection and electroporation; and the effect of repeated gene transfer by electroporation, were systematically investigated in the present study. The efficiencies of gene transfer by electroporation in normal and rodent models of diabetes were also evaluated. We found that electroporation used for non‐viral gene transfer could be repeated in the same place in the muscle, but the expression efficiency was closely related to the muscle damage. Increasing pulse times could enhance the efficiency of gene transfer with a lower strength of electric field. It was better to use a higher plasmid concentration than to use a larger dose of plasmid and repeated injection to achieve a high level of transgene expression. Optimal conditions varied in different animal models, being milder for diabetic mice than for normal mice, and it was also shown that the conditions that worked well on these small rodents were not necessarily suitable for larger animals. Our results provide a comprehensive view of the factors that affect the efficiency of gene transfer into skeletal muscle by electroporation. Copyright © 2005 John Wiley & Sons, Ltd.
Proliferation in the epidermis of chelonians and growth of the horny scutes
Authors: Alibardi Lorenzo
Published: May 2005

The proliferation of the epidermis in soft skin, claws, and scutes of the carapace and plastron in the tortoise (Testudo hermanni) and the turtle (Chrysemys picta) were studied using autoradiographic and immunocytochemical methods. During the growing season, a basal keratinocyte in the epidermis of soft skin and claws takes 5–9 days to migrate into the corneous layer. In the tortoise, during fall/winter (resting season) a few alpha‐keratin cells are produced in soft epidermis and hinge regions among scutes and occasional beta‐keratin cells in the outer scute surface. When growth is resumed in spring (growing season), cell proliferation is intense, mainly around hinge regions and tips of marginal scutes. No scute shedding occurs and numerous beta‐keratin cells are produced around the hinge regions, while alpha‐keratin cells disappear. Beta‐cells form a new thick corneous layer around the hinge regions, which constitute the growing rings of scutes. Beta‐keratin cells produced in more central parts of scutes maintain a homogeneous thickness of the corneous layer along the whole scute surface. In the turtle, a more complicated process of scute growth occurs than in the tortoise. At the end of the growing season (late fall) the last keratinocytes formed beneath the old stratum corneum of the outer scale surface and hinge regions produce more alpha‐ than beta‐keratin. These thin alpha‐keratin cells form a scission layer below the old stratum corneum, which extends from the hinge regions toward the center of scutes and the tip of marginal scutes. In the resting season (fall/winter) most cells remain within the germinative layer of the carapace and plastron and a few alpha‐cells move in 7–9 days into the corneous layer above hinge regions. In the following spring/summer (growing season) a new generation of beta‐keratin cells is produced beneath the scission layer from the hinge region and more central part of the scutes. The epidermis of the inner surface of scutes and hinge regions contains most of the cells incorporating thymidine and histidine, while the remaining outer scute surface is less active. It takes 5–9 days for a newly produced beta‐cell to migrate into the corneous layer. These cells form a new corneous layer that extends the whole scute surface underneath the maturing scission layer. The latter contains lipids and eventually flakes off, determining shedding of the above outer corneous layer in late spring or summer. J. Morphol. © 2005 Wiley‐Liss, Inc.
Postnatal ontogeny, population structure, and extinction of the giant moa Dinornis
Authors: Turvey Samuel T. , Holdaway Richard N.
Published: May 2005

Recent reinterpretation of the giant moa Dinornis as consisting of two sexually dimorphic allospecies permits thorough site‐by‐site investigation of the ontogeny and population biology of this genus. Analysis of subadult skeletal material from natural swamp sites in the North and South Islands of New Zealand forms the basis for recognition of growth series for each long bone element, characterized by sequential formation of fossulae in the femur and fusion of bones in the tibiotarsus and tarsometatarsus. Femora reached progressive developmental stages more rapidly than the other long bones, but all three elements reached maturity at about the same time. Patterns of bone fusion in Dinornis are more similar to those in Apteryx than in Struthio, and kiwi are recognized as a suitable developmental analog for interpreting moa ontogeny. Samples from Bell Hill Vineyard Swamp (South Island) and Makirikiri swamp (North Island) are interpreted as representing autochthonous moa populations; comparison with stages of kiwi long bone development suggests that Dinornis at these sites had high adult survivorship in strongly K‐selected populations, with 72.5–87.3% of individuals having achieved adult body mass, and 55.9–78.2% being sexually mature. The pattern of low fecundity and probable high longevity in both Dinornis species suggests that populations were vulnerable to loss of adults, primarily through hunting, rather than as a result of habitat destruction. J. Morphol. © 2005 Wiley‐Liss, Inc.
Statistical design of experiments as a tool in mass spectrometry
Authors: Riter Leah S. , Vitek Olga , Gooding Karen M. , Hodge Barry D. , Julian Randall K.
Published: May 2005

This Tutorial is an introduction to statistical design of experiments (DOE) with focus on demonstration of how DOE can be useful to the mass spectrometrist. In contrast with the commonly used one factor at a time approach, DOE methods address the issue of interaction of variables and are generally more efficient. The complex problem of optimizing data‐dependent acquisition parameters in a bottom‐up proteomics LC‐MS/MS analysis is used as an example of the power of the technique. Using DOE, a new data‐dependent method was developed that improved the quantity of confidently identified peptides from rat serum. Copyright © 2005 John Wiley & Sons, Ltd.
Protective effects of bone marrow stromal cell transplantation in injured rodent brain: Synthesis of neurotrophic factors
Authors: Chen Qin , Long Yan , Yuan Xiaoqing , Zou LingLong , Sun Jiao , Chen Shengdi , Perez‐Polo J. Regino , Yang Keyi
Published: May 2005

Several groups have suggested that transplantation of marrow stromal cells (MSCs) promotes functional recovery in animal models of brain trauma. Recent studies indicate that tissue replacement by this method may not be the main source of therapeutic benefit, as transplanted MSCs have only limited ability to replace injured central nervous system (CNS) tissue. To gain insight into the mechanisms responsible for such effects, we systematically investigated the therapeutic potential of MSCs for treatment of brain injury. Using in vitro studies, we detected the synthesis of various growth factors, including nerve growth factor (NGF), brain‐derived neurotrophic factor (BDNF), glial cell line‐derived neurotrophic factor (GDNF), and neurotrophin‐3 (NT‐3). Enzyme‐linked immunosorbent assay (ELISA) demonstrated that MSCs cultured in Dulbecco's modified Eagle medium (DMEM) produced substantial amounts of NGF for at least 7 weeks, whereas the levels of BDNF, GDNF and NT‐3 remained unchanged. In studies in mice, after intraventricular injection of MSCs, NGF levels were increased significantly in cerebrospinal fluid by ELISA, confirming our cell culture results. Further studies showed that treatment of traumatic brain injury with MSCs could attenuate the loss of cholinergic neuronal immunostaining in the medial septum of mice. These studies demonstrate for the first time that by increasing the brain concentration of NGF, intraventricularly transplanted MSCs might play an important role in the treatment of traumatic brain injury. © 2005 Wiley‐Liss, Inc.
Demonstration of kynurenine aminotransferases I and II and characterization of kynurenic acid synthesis in cultured cerebral cortical neurons
Authors: Rzeski Wojciech , Kocki Tomasz , Dybel Anna , Wejksza Katarzyna , Zdzisińska Barbara , Kandefer‐Szerszeń Martyna , Turski Waldemar A. , Okuno Etsuo , Albrecht Jan
Published: May 2005

The present study characterizes the synthesis of kynurenic acid (KYNA) from exogenously added kynurenine and its regulation by extrinsic factors, in cultured cerebral cortical neurons and, for comparison, in astrocytes incubated under identical conditions. The neuronal culture showed positive immunostaining for both kynurenic acid aminotransferase (KAT) isoforms I and II. Neurons synthesized KYNA at a rate about 2.3 times higher than astrocytes. Neuronal, but not astrocytic, KYNA synthesis was lowered ∼30% by ionotropic glutamate receptor agonists [(R,S)‐3‐hydroxy‐5‐methoxyloxasole‐4‐propionic acid (AMPA; 100 μM) and N‐methyl‐D‐aspartic acid (NMDA; 100 μM)] and depolarizing agents [KCl (50 mM) and 4‐aminopyridine (4‐AP; 10 μM)]. Neuronal and astrocytic synthesis alike were vulnerable to inhibition exerted by the aminotransferase inhibitor aminooxyacetic acid (AOAA), glutamate (IC50: 31 and 85 μM, respectively), substrates of the L‐amino transport system [leucine (Leu); IC50: 19 and 42 μM, respectively] and 2‐aminobicyclo[2,2,1]heptane‐2‐carboxylic acid (BCH; IC50: 19 and 28 μM, respectively). Glutamine (Gln), which is a metabolic precursor of glutamate in astrocytes and L‐system substrate in both cell types, inhibited KYNA synthesis both in neurons and in astrocytes (IC50: 268 and 318 μM, respectively). α‐Ketoisocaproic acid (KIC), a Leu transamination product that is produced mainly in astrocytes and shuttled to neurons to modulate intraneuronal concentration of glutamate, stimulated KYNA synthesis in neurons but did not affect the synthesis in astrocytes. In conclusion, this study is the first to demonstrate active, regulation‐prone KYNA synthesis in neurons. © 2005 Wiley‐Liss, Inc.
Albumin stimulates monocyte chemotactic protein‐1 expression in rat embryonic mixed brain cells
Authors: Calvo Charles‐Félix , Amigou Edwige , Tencé Martine , Yoshimura Teizo , Glowinski Jacques
Published: May 2005

Albumin, a blood protein absent from the adult brain in physiological situations, can be brought into contact with brain cells during development or, in adult, following breakdown of the blood–brain barrier occurring as a result of local inflammation. In the present study, we show that ovalbumin and albumin induce the release of monocyte chemotactic protein 1 (MCP‐1/CCL2) from rat embryonic mixed brain cells. A short‐term exposure to ovalbumin during the cell dissociation procedure is sufficient to generate MCP‐1 mRNA. A comparable effect is observed when the cells are incubated for 4 hr with ovalbumin or rat albumin, while MCP‐1 messengers are barely detectable following bovine albumin exposure. The amount of MCP‐1 protein measured in 4 hr‐supernatants of albumin‐treated cells followed the same albumin‐inducing pattern as that of MCP‐1 mRNA, while all albumins tested induced MCP‐1 protein after a 17 hr‐incubation period. The albumin‐induced MCP‐1 production is significantly inhibited in calphostin C‐treated cells, suggesting the implication of a protein kinase C‐dependent signaling pathway. This MCP‐1‐inducing activity is maintained after a lipid extraction procedure but abolished by proteinase K or trypsin treatments of albumin. The MCP‐1 secretion following albumin contact with nervous cells could thus interfere, by chemotactic gradient formation, with the brain infiltration program of blood‐derived cells during development or brain injury. © 2005 Wiley‐Liss, Inc.
A study of the effects of phototherapy dose interval on photobiomodulation of cell cultures
Authors: Brondon Philip , Stadler Istvan , Lanzafame Raymond J.
Published: May 2005

Background and Objectives

Dosimetry and treatment frequency are controversial phototherapy issues. Efficacy of dose fractionation on photobiomodulation was evaluated in vitro.

Study Design/Materials and Methods

Human HEP‐2 and murine L‐929 cell lines were cultured in complete DMEM media. Photoradiation (670 nm, 5 J/cm2/treatment, 50 J/cm2 total energy delivery), was performed varying treatments per 24 hour period: Group I (Controls)—0, Group II‐1/d, Group III‐2/d, Group IV‐4/d. Cell proliferation was measured using Cyquant® (fluorescent DNA content) and MTT (3‐(4, 5‐dimethylthiazol‐2‐yl)‐2, 5‐diphenyl tetrasolium bromide) assays for 240 hours post therapy. A proliferation index: PI = (#Cells Experimentalt ÷ #Cells Controlt) was computed.

Results

MTT assay results demonstrated maximal response in Group III (P < 0.05, n = 3). Cyquant® maxima occurred in HEP‐2 Groups II and III (P < 0.045) and L‐929 Group III (P < 0.091).

Conclusions

Cellular response to dose frequency varies. More frequent treatments (2/24 hours) increased metabolism and proliferation in both cell lines. Further investigation of dose fractionation in phototherapy is warranted. © 2005 Wiley‐Liss, Inc.
Solvent Vapor‐Induced Nanowire Formation in Poly(3‐hexylthiophene) Thin Films
Authors: Kim Do Hwan , Park Yeong Don , Jang Yunseok , Kim Sungsoo , Cho Kilwon
Published: May 2005

Summary: Nanowire lengths and length‐to‐width aspect ratios in regioregular poly(3‐hexylthiophene) (P3HT) were simply controlled through changes in the solvent vapor pressure during solidification. It is demonstrated that the nanowires grew by rod‐to‐rod association, in which the molecular long axis of the P3HT chains appeared to be well‐oriented parallel to the silicon substrate (Si/SiOx). The formation of the nanowires took place by one dimensional self‐assembly, governed by π‐π stacking of the P3HT units.

TEM high contrast images showing P3HT nanowires fabricated by spin‐coating under a solvent vapor pressure.

magnified image

TEM high contrast images showing P3HT nanowires fabricated by spin‐coating under a solvent vapor pressure.
Gait analysis detects early changes in transgenic SOD1(G93A) mice
Authors: Wooley Christine M. , Sher Roger B. , Kale Ajit , Frankel Wayne N. , Cox Gregory A. , Seburn Kevin L.
Published: May 2005

The effective treatment or cure of motoneuron disease will require understanding the disease processes that precede irreversible cell loss. To study these early stages, and to evaluate potential treatments in relevant animal models, requires a sensitive functional assay. To this end, we sought to determine whether the gait pattern of SOD1 transgenic mice changed prior to overt symptoms. Using a simplified video‐based approach we compared the treadmill gait of C57BL/6J and B6.SOD1 transgenic mice at 8 and 10 weeks of age. B6.SOD1 mice had significantly longer stride and stance times than controls by 8 weeks. Consistent with disease progression, hindpaw measures of B6.SOD1 mice showed larger changes than front paws. Differences between control and B6.SOD1 mice increased at 10 weeks, but only because repeat testing caused habituation in control mice to a greater extent than in B6.SOD1 mice. Together the results demonstrate that simplified gait analysis is sensitive to early processes of motor system disease in mice. Muscle Nerve, 2005
X‐linked Emery–Dreifuss muscular dystrophy and vacuoles: An immunohistochemical characterization
Authors: Paradas Carmen , Márquez Celedonio , Gallardo Eduard , de Luna Noemí , Chinchón Isidoro , Recan Dominique , Jiménez María Dolores , Illa Isabel
Published: May 2005

We report a striking abundance of rimmed vacuoles in two brothers with X‐linked Emery–Dreifuss muscular dystrophy (X‐EDMD) confirmed by the absence of emerin at the muscular nuclear envelope and by genetic analysis showing a new 2‐bp deletion in exon 6 of the STA gene at the Xq28 region. Immunohistochemical analysis of the vacuoles revealed expression of dystrophin but not of merosin in the sarcolemma of rimmed vacuoles and absence of amyloid and membrane attack complex (MAC) deposition either in vacuoles or muscle fibers. The presence of rimmed vacuoles can be a histopathological finding in X‐EDMD, and the diagnosis should not be excluded in clinically well‐defined EDMD patients because of this finding. Muscle Nerve, 2005
Intraneural microstimulation of motor axons in the study of human single motor units
Authors: McNulty Penelope A. , Macefield Vaughan G.
Published: May 2005

Single motor unit activity has been studied in depth since the first intramuscular electrodes were developed more than 70 years ago. Many techniques have been combined or used in isolation since then. Intraneural motor axon microstimulation allows the detailed study of single motor units in awake human subjects in a manner most analogous to that used in reduced animal preparations. A microelectrode, inserted percutaneously into a peripheral nerve, stimulates the axon of a single α‐motoneuron at a site remote from the contracting muscle, allowing detailed analyses of the contractile properties of a single motor unit in an otherwise quiescent muscle, that is, without interference of simultaneously active motor units or the presence of an electrode within the muscle. The methods and results obtained using this technique are described and compared to those of other studies of single motor units in human subjects. Differences have been found between human and animal motor units and between motor units of various muscles. Studying human and animal motor units using an analogous technique provides insight into the interpretation of human data when results differ from animal data, and when human motor units cannot be examined in the same way, or at a similar level of detail, as animal motor units. Muscle Nerve, 2005
Modifiability of abnormal isometric elbow and shoulder joint torque coupling after stroke
Authors: Ellis Michael D. , Holubar Bradley G. , Acosta Ana Maria , Beer Randall F. , Dewald Julius P. A.
Published: May 2005

Unlike individuals with mild stroke, individuals with severe stroke are constrained to stereotypical movement patterns attributed to abnormal coupling of shoulder abductors with elbow flexors, and shoulder adductors with elbow extensors. Whether abnormal muscle coactivation and associated joint torque patterns can be changed in this population is important to determine given that it bears on the development of effective rehabilitation interventions. Eight subjects participated in a protocol that was designed to reduce abnormal elbow/shoulder joint torque coupling by training them to generate combinations of isometric elbow and shoulder joint torques away from the constraining patterns. After training, subjects demonstrated a significant reduction in abnormal torque coupling and a subsequent significant increase in ability to generate torque patterns away from the abnormal pattern. We suggest the rapid time‐course of these changes reflects a residual capacity of the central nervous system to adapt to a novel behavioral training environment. Muscle Nerve, 2005
Effect of body mass index on ulnar nerve conduction velocity, ulnar neuropathy at the elbow, and carpal tunnel syndrome
Authors: Landau Mark E. , Barner Kristen C. , Campbell William W.
Published: May 2005

Both high and low body mass index (BMI) have been reported as risk factors for ulnar neuropathy at the elbow (UNE), and a high BMI as a risk factor for carpal tunnel syndrome (CTS). To determine whether the extremes of BMI are risk factors for UNE or CTS, and whether BMI affects calculation of median and ulnar motor nerve conduction velocity (NCV), we retrospectively analyzed the electrodiagnostic records of control patients, UNE patients, and CTS patients. The BMI was calculated for 50 patients with a sole diagnosis of UNE and compared to the BMI of 50 patients with CTS and 50 control subjects. The mean BMIs were 25.9 ± 4.4, 30.1 ± 5.5, and 28.3 ± 5.6 for the UNE, CTS, and controls, respectively. By one‐way analysis of variance, the difference in BMI between the UNE patients and the normal patients was significant (P < 0.01). In the control groups, increasing BMI directly correlated with increasing ulnar motor NCV across the elbow but not with forearm NCV. Across‐elbow (AE) ulnar motor NCV may be falsely increased in patients with a high BMI, probably due to distance measurement factors. Not only do relatively slender individuals have comparatively slower AE ulnar NCVs, they are also at increased risk for developing UNE. Patients with a high BMI are at increased risk for CTS. Muscle Nerve, 2005
Bounds for the eigenvalues of block 2 × 2 Hermitian positive‐definite matrices
Authors: Kolotilina L. Yu.
Published: May 2005

Lower and upper conditional bounds for the eigenvalues of a Hermitian positive‐definite block 2 × 2 matrix, describing their closest possible location, are derived. The notions of partially and totally optimal spectra are introduced, and several equivalent characterizations of matrices with partially and totally optimal spectra are presented. It is shown that the block 2 × 2 block Jacobi scaled matrices and the so‐called equilibrated matrices have totally optimal spectra. Copyright © 2005 John Wiley & Sons, Ltd.
A finite volume element method for a non‐linear elliptic problem
Authors: Chatzipantelidis P. , Ginting V. , Lazarov R. D.
Published: May 2005

We consider a finite volume discretization of second‐order non‐linear elliptic boundary value problems on polygonal domains. Using relatively standard assumptions we show the existence of the finite volume solution. Furthermore, for a sufficiently small data the uniqueness of the finite volume solution may also be deduced. We derive error estimates in H1‐, L2‐ and L∞‐norm for small data and convergence in H1‐norm for large data. In addition a Newton's method is analysed for the approximation of the finite volume solution and numerical experiments are presented. Copyright © 2005 John Wiley & Sons, Ltd.