bluuebunny/crossref_metadata_2025 huggingface dataset:
Enhanced cartilage tissue engineering by sequential exposure of chondrocytes to FGF‐2 during 2D expansion and BMP‐2 during 3D cultivation
Authors: Martin Ivan , Suetterlin R. , Baschong W. , Heberer M. , Vunjak‐Novakovic G. , Freed L.E.
Published: August 2002

Bovine calf articular chondrocytes, either primary or expanded in monolayers (2D) with or without 5 ng/ml fibroblast growth factor‐2 (FGF‐2), were cultured on three‐dimensional (3D) biodegradable polyglycolic acid (PGA) scaffolds with or without 10 ng/ml bone morphogenetic protein‐2 (BMP‐2). Chondrocytes expanded without FGF‐2 exhibited high intensity immunostaining for smooth muscle α‐actin (SMA) and collagen type I and induced shrinkage of the PGA scaffold, thus resembling contractile fibroblasts. Chondrocytes expanded in the presence of FGF‐2 and cultured 6 weeks on PGA scaffolds yielded engineered cartilage with 3.7‐fold higher cell number, 4.2‐fold higher wet weight, and 2.8‐fold higher wet weight glycosaminoglycan (GAG) fraction than chondrocytes expanded without FGF‐2. Chondrocytes expanded with FGF‐2 and cultured on PGA scaffolds in the presence of BMP‐2 for 6 weeks yielded engineered cartilage with similar cellularity and size, 1.5‐fold higher wet weight GAG fraction, and more homogenous GAG distribution than the corresponding engineered cartilage cultured without BMP‐2. The presence of BMP‐2 during 3D culture had no apparent effect on primary chondrocytes or those expanded without FGF‐2. In summary, the presence of FGF‐2 during 2D expansion reduced chondrocyte expression of fibroblastic molecules and induced responsiveness to BMP‐2 during 3D cultivation on PGA scaffolds. © 2001 Wiley‐Liss, Inc.
Comparative study of relativistic density functional methods applied to actinide species AcO22+ and AcF6 for Ac = U, Np
Authors: García‐Hernández Maite , Lauterbach Christa , Krüger Sven , Matveev Alexei , Rösch Notker
Published: August 2002

A two‐component relativistic density functional method based on the Douglas‐Kroll‐Hess transformation has been applied to the actinyls and hexafluorides of U and Np. All‐electron scalar relativistic calculations as well as calculations including spin‐orbit interaction have been compared to results obtained with a pseudopotential approach. In addition, several exchange‐correlation potentials have been applied to examine their performance for the bond lengths and vibrational frequencies of the title compounds. The calculations confirm the well‐known accuracy of the LDA approach for geometries and frequencies, which is corroborated for the hexafluorides where gas phase experimental data are available. Comparison with results of accurate wave function based methods provides further confirmation of this finding. Gradient‐corrected functionals tend to overestimate bond lengths and underestimate frequencies also for actinide compounds. The results obtained with Stoll‐Preuss (small core) effective core potentials agree very well with those of all‐electron calculations, while calculations with Hay‐Martin large core pseudopotentials are somewhat less accurate. For all molecules and properties considered, spin‐orbit effects have been found negligible concomitant with the closed‐shell electronic structure of the U(VI) compounds and the open‐shell situation of the Np(VI) compounds with a single valence f electron. © 2002 Wiley Periodicals, Inc. J Comput Chem 23: 834–846, 2002
Multiple grid methods for classical molecular dynamics
Authors: Skeel Robert D. , Tezcan Ismail , Hardy David J.
Published: August 2002

Presented in the context of classical molecular mechanics and dynamics are multilevel summation methods for the fast calculation of energies/forces for pairwise interactions, which are based on the hierarchical interpolation of interaction potentials on multiple grids. The concepts and details underlying multigrid interpolation are described. For integration of molecular dynamics the use of different time steps for different interactions allows longer time steps for many of the interactions, and this can be combined with multiple grids in space. Comparison is made to the fast multipole method, and evidence is presented suggesting that for molecular simulations multigrid methods may be superior to the fast multipole method and other tree methods. © 2002 Wiley Periodicals, Inc. J Comput Chem 23: 673–684, 2002
Computation of the physio‐chemical properties and data mining of large molecular collections
Authors: Cheng Ailan , Diller David J. , Dixon Steven L. , Egan William J. , Lauri George , Merz Kenneth M.
Published: August 2002

Very large data sets of molecules screened against a broad range of targets have become available due to the advent of combinatorial chemistry. This information has led to the realization that ADME (absorption, distribution, metabolism, and excretion) and toxicity issues are important to consider prior to library synthesis. Furthermore, these large data sets provide a unique and important source of information regarding what types of molecular shapes may interact with specific receptor or target classes. Thus, the requirement for rapid and accurate data mining tools became paramount. To address these issues Pharmacopeia, Inc. formed a computational research group, The Center for Informatics and Drug Discovery (CIDD).* In this review we cover the work done by this group to address both in silico ADME modeling and data mining issues faced by Pharmacopeia because of the availability of a large and diverse collection (over 6 million discrete compounds) of drug‐like molecules. In particular, in the data mining arena we discuss rapid docking tools and how we employ them, and we describe a novel data mining tool based on a 1D representation of a molecule followed by a molecular sequence alignment step. For the ADME area we discuss the development and application of absorption, blood–brain barrier (BBB) and solubility models. Finally, we summarize the impact the tools and approaches might have on the drug discovery process. © 2002 Wiley Periodicals, Inc. J Comput Chem 23: 172–183, 2002
Comparison of the Peabody Picture Vocabulary Test—Third Edition and Wechsler Adult Intelligence Scale—Third Edition with university students
Authors: Bell Nancy L. , Lassiter Kerry S. , Matthews T. Darin , Hutchinson Melody B.
Published: August 2002

This study examined the relationship between the Peabody Picture Vocabulary Test—Third Edition (PPVT‐III) and Wechsler Adult Intelligence Scale—Third Edition (WAIS‐III) using 40 adults who ranged in age from 18 to 41 (mean age of 22 years). Participants were administered the PPVT‐III and WAIS‐III in counterbalanced fashion to control for order effects. Results revealed that the PPVT‐III score was related to the WAIS‐III Verbal IQ (VIQ) and Full Scale IQ (FSIQ) scores but unrelated to the Performance IQ (PIQ) score. In addition, analyses indicated that, while there were no significant differences between the PPVT‐III score and WAIS‐III mean FSIQ and PIQ scores, the PPVT‐III mean score was significantly lower than the WAIS‐III VIQ. Further analysis indicated that the PPVT‐III adequately estimated WAIS‐III FSIQ and VIQ scores for participants who were classified as Average or High Average on the WAIS‐III. However, for participants in the Superior range, the PPVT‐III tended to underestimate FSIQ and VIQ scores by approximately 10 points. © 2001 John Wiley & Sons, Inc. J Clin Psychol 57: 417–422, 2001.
Naming and verbal memory skills in adults with Attention Deficit Hyperactivity Disorder and Reading Disability
Authors: Rashid Fontina L. , Morris Mary K. , Morris Robin
Published: August 2002

Research suggests that children with Attention Deficit Hyperactivity Disorder (ADHD) and Reading Disability (RD) can be differentiated based on their performance on measures of naming and verbal memory. It is not known whether this same pattern characterizes adults with these disorders. In this study, adults with and without ADHD and RD were compared on naming and verbal memory abilities. Results did not support the hypothesis that adults with ADHD and RD are significantly different from each other, or from a contrast group, on naming and verbal memory measures after controlling for Full‐Scale IQ (FSIQ). These results were replicated across two sets of classification criteria used to identify diagnostic groups. Factors that could account for the inconsistency in research findings with children and adult subjects are discussed. © 2001 John Wiley & Sons, Inc. J Clin Psychol 57: 829–838, 2001.
Molecular markers and determinants of prostate cancer metastasis
Authors: Gopalkrishnan Rahul V. , Kang Dong‐chul , Fisher Paul B.
Published: August 2002

Although intensely studied, the molecular and biochemical determinants of prostate cancer development and progression remain ill‐defined. Moreover, current markers and methodologies cannot distinguish between a tumor that will remain indolent and not impinge on patient survival, versus a tumor with aggressive traits culminating in metastatic spread and death. Once prostate cancer is confirmed the most significant threat to a patient's survival and quality of life involves tumor metastasis. Radical surgery notwithstanding, prostate cancer accounts for 10% of all cancer‐related deaths primarily arising through development of metastasis. Metastasis markers demonstrating an acceptable level of reliability are an obvious necessity if disproportionate and costly treatment is to be avoided and a reasonably accurate determination of clinical prognosis and measure of successful response to treatment is to be made. Therapeutic strategies that specifically inhibit metastatic spread are not presently possible and may not become available in the immediate future. This is because, while localized tumorigenesis has been relatively amenable to detection, analysis and treatment, metastasis remains a relatively undefined, complex and underexplored area of prostate cancer research. New findings in the field such subclasses of genes called metastasis suppressors and cancer progression suppressors, have opened up exciting avenues of investigation. We review current methodological approaches, model experimental systems and genes presently known or having potential involvement in human prostate cancer metastasis. © 2001 Wiley‐Liss, Inc.
Overexpression of the V3 variant of versican alters arterial smooth muscle cell adhesion, migration, and proliferation in vitro
Authors: Lemire Joan M. , Merrilees Mervyn J. , Braun Kathleen R. , Wight Thomas N.
Published: August 2002

Versican is an extracellular matrix proteoglycan produced by many cells. Although versican is generally known as a large chondroitin sulfate proteoglycan (CSPG), the smallest splice variant, V3, consists only of the amino‐ and carboxy‐terminal globular domains and is therefore predicted to be a small glycoprotein, lacking CS chains. The large size, negative charge, and ability of versican variants to form pericellular coats with hyaluronan are responsible for many of its effects. V3, lacking the large size and high charge density, but retaining the hyaluronan‐binding domain of the larger isoforms, may have different effects on cell phenotype. To determine whether V3 alters cell phenotype, Fisher rat arterial smooth muscle cells (ASMCs), which express the larger CSPG versican splice forms (V0 and V1) were retrovirally transduced with the rat V3 cDNA. Northern analysis for versican RNAs confirmed that cells transduced with V3 retrovirus, but not cells tranduced with the empty vector, expressed RNA of the size expected for V3/neor bicistronic RNA. V3 overexpressing cells were more spread on tissue culture plastic, had a smaller length‐to‐breadth ratio and were more resistant to release from the culture dish by trypsin. Interference reflection microscopy of sparsely plated cells showed larger areas of close contact between the V3 expressing cells and the coverslip, in comparison to control cells. Focal contacts in the periphery of V3 expressing cells were larger. Growth and migration studies revealed that V3 transduced cells grow slower and migrate a shorter distance in a scratch wound assay. The increased adhesion and the inhibition of migration and proliferation resulting from V3 overexpression are the opposites of the known and predicted effects of the other variants of versican. V3 may exert these effects through changes in pericellular coat formation, either by competing with larger isoforms for hyaluronan‐binding, or by altering other components of the pericellular matrix. J. Cell. Physiol. 190: 38–45, 2002. © 2002 Wiley‐Liss, Inc.
Functional analysis of mouse hepatocytes differing in DNA content: Volume, receptor expression, and effect of IFNγ
Authors: Martin Nicola C. , McCullough Christian T. , Bush Peter G. , Sharp Linda , Hall Andrew C. , Harrison David J.
Published: August 2002

Polyploidy and binuclearity are characteristics of the mammalian liver. Increasing polyploidisation occurs with age and after administration of various drugs and chemicals. This study was designed to examine the function of ploidy by addressing several questions: (1) Does the increase in size of polyploid hepatocytes have any physiological function by altering surface receptor expression such as intercellular adhesion molecule‐1 (ICAM‐1, CD54) or IFNγR? and (2) Do polyploid cells respond differently to inflammatory cytokines such as interferon gamma (IFNγ)? We have developed a method to accurately measure the volume of live isolated hepatocytes using confocal microscopy and image analysis. Using flow cytometry, we have shown that the expression of ICAM‐1 increases with increasing DNA content and IFNγR is not detectable on isolated mouse hepatocytes. Diploid (2n), tetraploid (4n) and octoploid (8n) hepatocytes were found to be equally susceptible to IFNγ‐induced apoptosis in vitro. Although the function of polyploidy remains unanswered, we have described some of the characteristics of polyploidy in isolated hepatocytes and in vitro. J. Cell. Physiol. 191: 138–144, 2002. © 2002 Wiley‐Liss, Inc.
Regulation of cell proliferation by Smad proteins
Authors: Ten Dijke Peter , Goumans Marie‐José , Itoh Fumiko , Itoh Susumu
Published: August 2002

Transforming growth factor‐β (TGF‐β) family members which include TGF‐βs, activins, and bone morphogenetic proteins (BMPs) regulate a broad spectrum of biological responses on a large variety of cell types. TGF‐β family members initiate their cellular responses by binding to distinct receptors with intrinsic serine/threonine kinase activity and activation of specific downstream intracellular effectors termed Smad proteins. Smads relay the signal from the cell membrane to the nucleus, where they affect the transcription of target genes. Smad activation, subcellular distribution, and stability have been found to be intricately regulated and a broad array of transcription factors have been identified as Smad partners. Important activities of TGF‐β are its potent anti‐mitogenic and pro‐apoptotic effects that, at least in part, are mediated via Smad proteins. Escape from TGF‐β/Smad‐induced growth inhibition and apoptosis is frequently observed in tumors. Certain Smads have been found to be mutated in specific types of cancer and gene ablation of particular Smads in mice has revealed increased rate of tumorigenesis. In late stage tumors, TGF‐β has been shown to function as a tumor promoter. TGF‐β can stimulate the de‐differentiation of epithelial cells to malignant invasive and metastatic fibroblastic cells. Interestingly, TGF‐β may mediate these effects directly on tumor cells via subverted Smad‐dependent and/or Smad‐independent pathways. J. Cell. Physiol. 191: 1–16, 2002. © 2002 Wiley‐Liss, Inc.
Cdc7 kinase complex: A key regulator in the initiation of DNA replication
Authors: Masai Hisao , Arai Ken‐Ichi
Published: August 2002

DNA replication results from the action of a staged set of highly regulated processes. Among the stages of DNA replication, initiation is the key point at which all the G1 regulatory signals culminate. Cdc7 kinase is the critical regulator for the ultimate firing of the origins of initiation. Cdc7, originally identified in budding yeast and later in higher eukaryotes, forms a complex with a Dbf4‐related regulatory subunit to generate an active kinase. Genetic evidence in mammals demonstrates essential roles for Cdc7 in mammalian DNA replication. Mini‐chromosome maintenance protein (MCM) is the major physiological target of Cdc7. Genetic studies in yeasts indicate additional roles of Cdc7 in meiosis, checkpoint responses, maintenance of chromosome structures, and repair. The interplay between Cdc7 and Cdk, another kinase essential for the S phase, is also discussed. J. Cell. Physiol. 190: 287–296, 2002. © 2002 Wiley‐Liss, Inc.
Hsp90: Chaperoning signal transduction
Authors: Richter Klaus , Buchner Johannes
Published: August 2002

Hsp90 is an ATP dependent molecular chaperone involved in the folding and activation of an unknown number of substrate proteins. These substrate proteins include protein kinases and transcription factors. Consistent with this task, Hsp90 is an essential protein in all eucaryotes. The interaction of Hsp90 with its substrate proteins involves the transient formation of multiprotein complexes with a set of highly conserved partner proteins. The specific function of each component in the processing of substrates is still unknown. Large ATP‐dependent conformational changes of Hsp90 occur during the hydrolysis reaction and these changes are thought to drive the chaperone cycle. Natural inhibitors of the ATPase activity, like geldanamycin and radicicol, block the processing of Hsp90 substrate proteins. As many of these substrates are critical elements in signal transduction, Hsp90 seems to introduce an additional level of regulation. © 2001 Wiley‐Liss, Inc.
Study on the enzymatic hydrolysis of racemic methyl ibuprofen ester
Authors: Madhav M Venu , Ching C B
Published: August 2002

The hydrolysis of racemic methyl ibuprofen ester in the presence of lipase from Candida rugosa was investigated in shake flasks. Experiments were performed to study the effect of temperature, pH and shaking speed on the reaction rate. Different hydrophobic co‐solvents were screened for the highest reaction rate and the presence of enzyme inhibition by substrate and products was examined. A kinetic expression was then proposed to describe the reaction. Kinetic parameters were determined for the optimum operating conditions and the proposed model was verified with the experimental results. Next, this reaction was scaled up to a fed batch stirred tank reactor. Batch reactor and fed batch reactor configurations were compared for better conversions. The effects of aqueous phase hold‐up, substrate concentration and feed flow rate on the conversion of the reaction were also studied. Higher conversions were obtained in a fed batch reactor when compared with the batch reactor. In the fed batch reactor, increased conversions were observed with lower feed flowrates and high aqueous phase hold‐up.

© 2001 Society of Chemical Industry
Kinetic studies of removal of ammonia from seawater by ozonation
Authors: Tanaka Junko , Matsumura Masatoshi
Published: August 2002

A simplified mathematical model for the estimation of the rate of removal of ammonia during the ozonation of seawater was developed, considering the kinetics of the interactions between ozone and both the chloride and bromide ions in seawater during oxidant formation and oxidation of ammonia. The model also considered effects of the rate of ozone mass transfer and the pH. The rates of removal of ammonia in the pH range between 6.5 and 9.0 coincided well with the values estimated by this model. It was also shown that stripping of ammonia during air/ozone treatment considerably affected the rate of removal of ammonia when the pH was higher than 8.5. The chloride ion, which is the dominant ion in seawater, showed a negligible contribution to the oxidation of ammonia during ozonation. The formation of BrO3−, classified as a carcinogen did not occur as long as ammonia was still present.

© 2002 Society of Chemical Industry
Ultrastructural studies define soluble macromolecular, particulate, and cellular transendothelial cell pathways in venules, lymphatic vessels, and tumor‐associated microvessels in man and animals
Authors: Feng Dian , Nagy Janice A. , Dvorak Harold F. , Dvorak Ann M.
Published: August 2002

We present de novo studies and review published efforts from our laboratory, spanning 12 years (from 1988 to 2000), where we have used ultrastructural approaches to study the functional anatomy of the microvasculature in man and animals in health and disease. These efforts have defined a new endothelial cell organelle, termed the vesiculo‐vacuolar organelle (VVO), which participates in the regulated transendothelial cell passage of soluble macromolecules. The studies defining this organelle utilized ultrathin serial sections, three‐dimensional computer‐assisted reconstructions, and ultrastructural electron‐dense tracers to establish luminal to abluminal transendothelial cell continuity of VVOs. Commonality of VVOs and caveolae is suggested by the ultrastructural anatomy of individual units of VVOs and caveolae, the presence of caveolin in both structures, and a mathematical analysis of morphometric data, all of which suggest that VVOs form from fusions of individual size units equivalent to vesicles of caveolar size. Ultrastructural studies have localized potent permeability factors and their specific receptors to VVOs in in vivo tumor and allergic inflammation models. Regulation of permeability through VVOs has been quantified and shown to be increased in tumor microvessels and in control vessels exposed to potent permeability‐inducing mediators. The transendothelial cell passage of particulate macromolecules occurs by vacuolar transport in tumor vessels; in permeability factor‐exposed control vessels, colloidal carbon traversed endothelial cells via the development of pores that did not communicate with or disrupt intercellular junctions by gap formation. Serial section and computer‐assisted reconstructions established these findings and suggested the possible development of transendothelial cell pores from VVOs. Serial sectioning and computer‐assisted three‐dimensional reconstructions of ultrastructural samples of an acute inflammation model revealed a transendothelial cell traffic route for motile neutrophils and platelets in the absence of classical ultrastructural criteria for regulated secretion from either cell. Microsc. Res. Tech. 57:289–326, 2002. © 2002 Wiley‐Liss, Inc.
Role of microglia in glioma biology
Authors: Badie Behnam , Schartner Jill
Published: August 2002

Microglia, a type of differentiated tissue macrophage, are considered to be the most plastic cell population of the central nervous system (CNS). In response to pathological conditions, resting microglia undergo a stereotypic activation process and become capable of phagocytosis, antigen presentation, and lymphocyte activation. Considering their immune effector function, it is not surprising to see microglia accumulation in almost every CNS disease process, including malignant brain tumors or malignant gliomas. Although the function of these cells in CNS inflammatory processes is being studied, their role in malignant glioma biology remains unclear. On one hand, microglia may represent a CNS anti‐tumor response, which is inactivated by local secretion of immunosuppressive factors by glioma cells. On the other hand, taking into account that microglia are capable of secreting a variety of immunomodulatory cytokines, it is possible that they are attracted by gliomas to promote tumor growth. A better understanding of microglia‐glioma interaction will be helpful in designing novel immune‐based therapies against these fatal tumors. Microsc. Res. Tech. 54:106–113, 2001. © 2001 Wiley‐Liss, Inc.
Drosophila as a focus in olfactory research: Mapping of olfactory sensilla by fine structure, odor specificity, odorant receptor expression, and central connectivity
Authors: Stocker Reinhard F.
Published: August 2002

This review intends to integrate recent data from the Drosophila olfactory system into an up‐to‐date account of the neuronal basis of olfaction. It focuses on (1) an electron microscopic study that mapped a large proportion of fruitfly olfactory sensilla, (2) large‐scale electrophysiological recordings that allowed the classification of the odor response spectra of a complete set of sensilla, (3) the identification and expression patterns of candidate odorant receptors in the olfactory tissues, (4) central projections of neurons expressing a given odorant receptor, (5) an improved glomerular map of the olfactory center, and (6) attempts to exploit the larval olfactory system as a model of reduced cellular complexity. These studies find surprising parallels between the olfactory systems of flies and mammals, and thus underline the usefulness of the fruitfly as an olfactory model system. Both in Drosophila and in mammals, odorant receptor neurons appear to express only one type of receptor. Neurons expressing a given receptor are scattered in the olfactory tissues but their afferents converge onto a few target glomeruli only. This suggests that in both phyla, the periphery is represented in the brain as a chemotopic map. The major difference between mammals and fruitflies refers to the numbers of receptors, neurons, and glomeruli, which are largely reduced in the latter, and particularly in larvae. Yet, if activated in a combinatorial fashion, even this small set of elements could allow discrimination between a vast array of odorants. Microsc. Res. Tech. 55:284–296, 2001. © 2001 Wiley‐Liss, Inc.
Effects of genic substitution at the pink‐eyed dilution locus on the proliferation and differentiation of mouse epidermal melanocytes in vivo and in vitro
Authors: Hirobe Tomohisa , Kawa Yoko , Mizoguchi Masako , Ito Shosuke , Wakamatsu Kazumasa
Published: August 2002

Cells positive to the dopa reaction (melanocytes) as well as to the combined dopa–premelanin reaction (melanoblasts and melanocytes) in the epidermis of C57BL/10JHir‐p/p (pink‐eyed dilution) mice were fewer and less reactive than in C57BL/10JHir (black, P/P) mice, suggesting that the proliferation and differentiation of p/p melanocytes are inhibited. To confirm the inhibitory effects of p gene on the proliferation and differentiation of epidermal melanocytes, we cultured epidermal cell suspensions of neonatal skins from P/P and p/p in a serum‐free medium. The proliferation and differentiation of p/p melanoblasts/melanocytes in primary culture were greatly inhibited as compared to P/P melanoblasts/melanocytes. The morphology of p/p melanoblasts/melanocytes cultured in melanocyte growth medium, though non‐pigmented, was similar to P/P melanocytes; namely, dendritic, polygonal, or epithelioid. About 8% of p/p cells cultured in melanocyte growth medium were positive to the dopa reaction, and about 25% were reactive to the combined dopa–premelanin reaction. Eumelanin content in p/p was extremely reduced compared to P/P. The immunocytochemical staining of p/p melanoblasts/melanocytes revealed that they are negative to tyrosinase, but reactive to tyrosinase‐related protein (TRP)‐1, TRP‐2, and c‐kit. However, the reactivities in p/p were lower than in P/P. Although the differentiation of p/p melanoblasts was not induced by endothelin (ET)‐1, ET‐2, and ET‐3, the proliferation of p/p melanoblasts was stimulated by them. These results suggest for the first time that p gene exerts its influence on the proliferative activities of mouse epidermal melanoblasts by affecting the regulatory mechanisms dependent on the function of ETs. J. Exp. Zool. 292:351–366, 2002. © 2002 Wiley‐Liss, Inc.
Preparation of metal ion buffers for biological experimentation: A methods approach with emphasis on iron and zinc
Authors: Aslamkhan Amy G. , Aslamkhan Abubakr , Ahearn Gregory A.
Published: August 2002

Transition metal ions are a challenge to study in physiology because of problems associated with solubility, oxidation, binding, and attaining appropriate free activities in solution. This review discusses these problems and potential ways of accommodating them. Special attention is given to iron and zinc ions, but many of the concepts can be applied for studying other transition metals. Selection of reagents appropriate for metal work (including water, salts, noncomplexing pH buffers) is briefly discussed. Calculation of the solubility product (Ksp) for common iron and zinc precipitates is covered, as well as techniques used to solubilize Fe3+ with organic chelates. Factors that affect Fe2+ oxidation are mentioned, and the use of ascorbate as a reducing agent is considered. Measurement of the rate of Fe2+ oxidation (or Fe3+ reduction) with the Fe2+ chromophores ferrozine and BPS is also discussed. Generation of a free metal ion activity through use of metal buffers (chelators) is discussed. Theoretical problems associated with this technique are explored, and selected shareware metal ion buffer calculators are described. Finally, techniques for measuring and minimizing nonspecific binding of iron and zinc ions to biological membranes are considered. J. Exp. Zool. 292:507–522, 2002. © 2002 Wiley‐Liss, Inc.
Differential induction of branchial carbonic anhydrase and NA+/K+ ATPase activity in the euryhaline crab, Carcinus maenas, in response to low salinity exposure
Authors: Henry Raymond P. , Garrelts Elizabeth E. , McCarty Melissa M. , Towle David W.
Published: August 2002

The time course of induction of activity of carbonic anhydrase (CA) and Na/K ATPase, two enzymes that are central to osmotic and ionic regulation in the eyryhaline green crab, Carcinus maenas, was measured in response to a transfer from 32 to 10 ppt salinity. CA activity was low in all gills in crabs acclimated to high salinity. Activity was induced in the posterior three gills (G6–G9) starting at 96 hr following transfer to low salinity, with activity peaking at seven post‐transfer. Na/K ATPase activity in posterior gills was already high in crabs acclimated to 32 ppt salinity, and it did not increase as a result of transfer to 10 ppt. Acclimation of crabs to hypersaline (40 ppt) conditions resulted in uniformly low levels of Na/K ATPase activity, and transfer from 40 ppt to 10 ppt stimulated a four‐fold induction of activity in the posterior gills that was evident by seven days of low salinity exposure. Low salinity stimulates the activity of both enzymes, but a different degree of salinity change appears to be necessary to cause the induction of each enzyme. The Na/K ATPase activity is already high at a salinity (32 ppt) at which the crab is still an osmotic and ionic conformer. CA activity, however, even when expressed in low levels, is still present in excess of what is needed to supply counterions at a rate adequate to match the rate of active ion transport. It is possible that two strategies exist for the regulation of these two enzymes that coincide with the crab’s intertidal and estuarine lifestyle: short‐term modulation of activity of highly expressed enzyme (Na/K ATPase) and long‐term modulation of enzyme concentration by changes in gene expression (CA). For all ranges of low salinity exposure, crabs undergo hemodilution, cell swelling, and subsequent cell volume readjustment as evidenced by the increase in concentration of TNPS in the hemolymph. This response takes place before the induction of enzyme activity, and it could serve as the initial signal in the induction pathway. J. Exp. Zool. 292:595–603, 2002. © 2002 Wiley‐Liss, Inc.
Sex reversal in Nile tilapia (Oreochromis niloticus) using a nonsteroidal aromatase inhibitor
Authors: Afonso Luís Orlando Bertolla , Wassermann Gustavo Javier , Terezinha de Oliveira Rosemari
Published: August 2002

Several studies have demonstrated that steroid hormones can influence sex differentiation in nonmammalian vertebrates and it has been hypothesized that male and female sex differentiation are driven by androgen and estrogen hormones, respectively. Estrogen biosynthesis is mediated by the steroidogenic enzyme cytochrome P450 aromatase, which converts androgens to estrogens. In the present study we examined the efficacy of a potent nonsteroidal aromatase inhibitor incorporated into the food, on sex reversal of Nile tilapia (Oreochromis niloticus) larvae. Nile tilapia larvae were divided in seven groups, which were fed with diets containing different amounts of the aromatase inhibitor Fadrozole (0, 50, 75 and 100 mg/kg) during 15 and 30 days, starting 9 days after hatching. Independent of the period, the proportion of males was significantly higher in the treated groups. Treatment with the highest doses (75 and 100 mg/kg) for 30 days produced 100% males. Histological examination revealed no differences in gonadal tissues between control males and treated fish. Furthermore, one intersex fish was identified in the group treated with 50 mg Fadrozole/kg for 30 days. This study reports that a 100% Nile tilapia male population can be obtained by suppressing aromatase activity and suggests that besides steroid hormones, nonsteroidal compounds, such as aromatase inhibitors, have potential for production of monosex population in tilapia. J. Exp. Zool. 290:177–181, 2001. © 2001 Wiley‐Liss, Inc.
Sustained delivery of therapeutic concentrations of human clotting factor IX – a comparison of adenoviral and AAV vectors administered in utero
Authors: Schneider Holm , Mühle Christiane , Marie Douar Anne , Waddington Simon , Jiang Qiu‐Jie , von der Mark Klaus , Coutelle Charles , Rascher Wolfgang
Published: August 2002

Background

Prenatal somatic gene therapy has been considered for genetic disorders presenting with morbidity at birth. Haemophilia is associated with an increased risk of catastrophic perinatal bleeding complications such as intracranial haemorrhage, which could be prevented by gene transfer in utero. Prenatal gene therapy may be more promising than postnatal treatment, as the fetus may be more amenable to uptake and integration of therapeutic DNA and the immaturity of its immune system may permit life‐long immune tolerance of the transgenic protein, thus avoiding the dominant problem in haemophilia treatment, the formation of inhibitory antibodies.

Methods

Adenovirus serotype 5‐derived or AAV serotype 2‐derived vectors carrying human clotting factor IX (hfIX) cDNA or a reporter gene were administered intramuscularly, intraperitoneally or intravascularly to late‐gestation mouse fetuses. Both vector types were evaluated with respect to the kinetics of hfIX delivery to the systemic circulation and possible immune responses against the vector or the transgene product.

Results

Mice treated in utero by intramuscular injection of an adenoviral vector carrying hfIX cDNA exhibited high‐level gene expression at birth and therapeutic – albeit continuously decreasing – plasma concentrations of hfIX over the entire 6 months of the study. Adenoviral vector spread to multiple organs was detected by polymerase chain reaction (PCR). Intramuscular, intraperitoneal or intravascular application of AAV vectors carrying hfIX cDNA led to much lower plasma concentrations of hfIX shortly after birth, which appeared to decline during the first month of life but stabilized in some of the mice at detectable levels. No signs of immune responses were found, either against the different viral vectors or against hfIX.

Conclusion

This study demonstrates for the first time that sustained systemic delivery of a therapeutic protein can be achieved by prenatal gene transfer. It thus shows the feasibility of gene therapy in utero and provides a basis for considering this concept as a preventive therapeutic strategy for haemophilia and perhaps also for other plasma protein deficiencies. Copyright © 2002 John Wiley & Sons, Ltd.
Use of a lentiviral flap vector for induction of CTL immunity against melanoma. Perspectives for immunotherapy
Authors: Firat Hüseyin , Zennou Véronique , Garcia‐Pons Francisco , Ginhoux Florent , Cochet Madeleine , Danos Olivier , Lemonnier François A. , Langlade‐Demoyen Pierre , Charneau Pierre
Published: August 2002

Background

A central triple‐stranded DNA structure created during HIV‐1 reverse transcription, the central flap, acts as a cis‐active nuclear import determinant of the HIV‐1 DNA genome. Insertion of the sequences responsible for formation of the central DNA flap into an HIV‐1‐derived vector strongly enhances its transduction efficiency.

Methods

HIV‐1 vectors with or without inclusion of the DNA flap and encoding the same melanoma polyepitope were constructed to transduce dendritic cells (DCs) and to evaluate their capacity for induction of melanoma‐specific cytotoxic T‐lymphocyte (CTL) responses ex vivo and in vivo.

Results

HIV‐1 vectors including the DNA flap transduced up to 100% of immature mouse and human DCs. Inoculation of HLA‐A2.1 transgenic mice with this flap vector elicited vigorous and multi‐specific long‐term anti‐melanoma CTL responses, whereas the parental vector lacking the flap sequence was less efficient. Furthermore, human DCs transduced ex vivo with the recombinant DNA flap vector displayed efficient multi‐specific primary human CTL responses against melanoma.

Conclusion

Lentiviral vectors including the DNA flap should be powerful tools both for active immunization and for the ex vivo priming of CTL for tumor immunotherapy. Copyright © 2002 John Wiley & Sons, Ltd.
Introducing psychology as an academic discipline in France: Théodule Ribot and the Collège de France (1888–1901)
Authors: Nicolas Serge , Charvillat Agnès
Published: August 2002

This paper describes the context in which the teaching of psychology as an autonomous discipline was introduced in France, and reproduces the first psychology lecture given in France by Théodule Ribot on 9 April 1888 at The Collège de France. In France, this recognition was delayed because of the negative influence of spiritualist philosophy. It took both the acknowledged status of a man (Ribot) and a minister's decision for this new type of teaching to be accepted in France. After describing the events that took place at the Collège de France and at the Academy of Moral and Political Sciences, we reproduce in full Ribot's inaugural lecture at the Collège, an important document for the history of French psychology. We conclude by describing the circumstances in which this teaching came to its end in 1901. © 2001 John Wiley & Sons, Inc.
The consultation practice of Northern NGOs: a study of British organizations in Guatemala
Authors: Lister Sarah
Published: August 2002

This paper presents the findings of research carried out in 1998–99 into the practices of seven British NGOs and solidarity organizations in consulting with local partner organizations in Guatemala. It considers this practice in the context of both the academic literature and the statements and intentions of the NGOs studied. The research found that while there was consultation on country‐specific issues, there was limited consultation on their broader programmes, strategies or advocacy work. Although the British NGOs claimed that they consulted for their own effectiveness, to strengthen local organizations and strengthen their partnership with these organizations, there was little evidence to suggest that their actual practice of consultation achieved these aims. Copyright © 2001 John Wiley & Sons, Ltd.
Information literacy needs of local staff in cross‐cultural development projects
Authors: McMahon Camille , Bruce Christine
Published: August 2002

Information literacy is about people's information practices in their information and communication technology (ICT) environment. Increasing access to ICTs to bridge the digital divide has implications for the information literacy needs of people in developing communities. The research described in this paper investigated development workers' perceptions of information literacy needs amongst local staff participating in community development projects in cross‐cultural situations. A phenomenographic approach was used to elicit five hierarchically related, qualitatively different understandings of information literacy needs within the development context. The results reveal possible directions for those involved in bringing ICTs into workplace settings within developing communities. Copyright © 2002 John Wiley & Sons, Ltd.
Explaining bank regulatory failure in Zambia
Authors: Maimbo Samuel Munzele
Published: August 2002

This paper discusses regulatory failure within the context of the 1995 and 1997–98 bank failures in Zambia. It starts by acknowledging the difficulties that exist in establishing credible techniques for measuring the quality of on‐going bank regulation and supervision performance and criterion for distinguishing between regulatory failure and regulatory success. It then proceeds to detail the choice of instruments the Bank of Zambia employed in responding to increasing financial distress amongst local banks. Using ‘excessive regulatory forbearance’ as an indicator of ‘regulatory failure’, the paper examines why regulatory forbearance occurred; why the Bank of Zambia failed to enforce the required corrective action in a timely and consistent manner, even at the risk of encouraging and permitting fraudulent and wrongful trading. Through in‐depth interviews and documentary analysis, the paper concludes that delays in regulatory decision‐making were not always the result of direct political interference, but rather bureaucratically institutionalised regulatory forbearance. Copyright © 2002 John Wiley & Sons, Ltd.
Synthesis of 3‐[(2S)‐azetidin‐2‐ylmethoxy]‐5‐[11C]‐methylpyridine, an analogue of A‐85380, via a Stille coupling
Authors: Karimi Farhad , Långström Bengt
Published: August 2002

3‐[(2S)‐azetidin‐2‐ylmethoxy]‐5‐[11C]‐methylpyridine (5d), which might be a novel ligand for nicotinic receptors, was synthesized via coupling [11C]iodomethane with tert‐butyl (2S)‐2‐({[5‐(trimethylstannyl)pyridin‐3‐yl]oxy}methyl) azetidine‐1‐carboxylate (4) at 80°C for 5 min with tri‐o‐tolylphosphine‐bound, unsaturated palladium(0), followed by deprotection using trifluoroacetic acid (TFA). The previous problem (solid‐phase extraction before injection on semi‐preparative LC) with automation of Stille coupling reactions has been overcome. In a typical experiment, 0.46 GBq of 5d was obtained from 5.2 GBq of [11C]iodomethane. The decay‐corrected radiochemical yield was 39% (based on the quantity [11C]iodomethane trapped). The synthesis time was 43 min from end of radionuclide production. During a production condition using 36 μAh of proton beam irradiation, a specific radioactivity of 50 GBq/μmol of the final product was obtained in biological buffer. Copyright © 2002 John Wiley & Sons, Ltd.
Evolution and mechanics of long jaws in butterflyfishes (Family Chaetodontidae)
Authors: Ferry‐Graham Lara A. , Wainwright Peter C. , Hulsey C. Darrin , Bellwood David R.
Published: August 2002

We analyzed the functional morphology and evolution of the long jaws found in several butterflyfishes. We used a conservative reanalysis of an existing morphological dataset to generate a phylogeny that guided our selection of seven short‐ and long‐jawed taxa in which to investigate the functional anatomy of the head and jaws: Chaetodon xanthurus, Prognathodes falcifer (formerly Chaetodon falcifer), Chelmon rostratus, Heniochus acuminatus, Johnrandallia nigrirostris, Forcipiger flavissimus, and F. longirostris. We used manipulations of fresh, preserved, and cleared and stained specimens to develop mechanical diagrams of how the jaws might be protruded or depressed. Species differed based on the number of joints within the suspensorium. We used high‐speed video analysis of five of the seven species (C. xanthurus, Chel. rostratus, H. acuminatus, F. flavissimus, and F. longirostris) to test our predictions based on the mechanical diagrams: two suspensorial joints should facilitate purely anteriorly directed protrusion of the lower jaw, one joint should allow less anterior protrusion and result in more depression of the lower jaw, and no joints in the suspensorium should constrain the lower jaw to simple ventral rotation around the jaw joint, as seen in generalized perciform fishes. We found that the longest‐jawed species, F. longirostris, was able to protrude its jaws in a predominantly anterior direction and further than any other species. This was achieved with little input from cranial elevation, the principal input for other known lower jaw protruders, and is hypothesized to be facilitated by separate modifications to the sternohyoideus mechanism and to the adductor arcus palatini muscle. In F. longirostris the adductor arcus palatini muscle has fibers oriented anteroposteriorly rather than medial‐laterally, as seen in most other perciforms and in the other butterflyfish studied. These fibers are oriented such that they could rotate the ventral portion of the quadrate anteriorly, thus projecting the lower jaw anteriorly. The intermediate species lack modification of the adductor arcus palatini and do not protrude their jaws as far (in the case of F. flavissimus) or in a purely anterior fashion (in the case of Chel. rostratus). The short‐jawed species both exhibit only ventral rotation of the lower jaw, despite the fact that H. acuminatus is closely related to Forcipiger. J. Morphol. 248:120–143, 2001. © 2001 Wiley‐Liss, Inc.
Functional morphology of the pectoral fins in bamboo sharks, Chiloscyllium plagiosum: Benthic vs. Pelagic station‐holding
Authors: Wilga Cheryl D. , Lauder George V.
Published: August 2002

Bamboo sharks (Chiloscyllium plagiosum) are primarily benthic and use their relatively flexible pectoral and pelvic fins to rest on and move about the substrate. We examined the morphology of the pectoral fins and investigated their locomotory function to determine if pectoral fin function during both benthic station‐holding and pelagic swimming differs from fin function described previously in leopard sharks, Triakis semifasciata. We used three‐dimensional kinematics and digital particle image velocimetry (DPIV) to quantify pectoral fin function in five white‐spotted bamboo sharks, C. plagiosum, during four behaviors: holding station on the substrate, steady horizontal swimming, and rising and sinking during swimming. During benthic station‐holding in current flow, bamboo sharks decrease body angle and adjust pectoral fin angle to shed a clockwise fluid vortex. This vortex generates negative lift more than eight times that produced during open water vertical maneuvering and also results in an upstream flow that pushes against the posterior surface of the pectoral fin to oppose drag. In contrast, there is no evidence of significant lift force in the wake of the pectoral fin during steady horizontal swimming. The pectoral fin is held concave downward and at a negative dihedral angle during steady horizontal swimming, promoting maneuverability rather than stability, although this negative dihedral angle is much less than that observed previously in sturgeon and leopard sharks. During sinking, the pectoral fins are held concave upward and shed a clockwise vortex with a negative lift force, while in rising the pectoral fin is held concave downward and sheds a counterclockwise vortex with a positive lift force. Bamboo sharks appear to sacrifice maneuverability for stability when locomoting in the water column and use their relatively flexible fins to generate strong negative lift forces when holding position on the substrate and to enhance stability when swimming in the water column. J. Morphol. 249:195–209, 2001. © 2001 Wiley‐Liss, Inc.