bluuebunny/crossref_metadata_2025 huggingface dataset:
The nature and dissemination of UHMWPE wear debris retrieved from periprosthetic tissue of THR
Authors: Elfick Alistair P. D. , Green Sarah M. , Krikler Steve , Unsworth Anthony
Published: March 2003

The role of wear debris in provoking joint replacement failure through bone resorption is now supported by much research. This study presents the analysis of 104 tissue samples using laser diffraction wear particle analysis in conjunction with standard histologic methods. The number and volume distributions were correlated to a range of joint and patient parameters. The median particle diameter by number was 0.69 μm. No particles smaller than 0.113 μm were resolved. No variation in terms of particle distribution was found among joint types. The ability of particles to migrate away from their point of origin was found to be inversely proportional to their size. The numbers of particles per gram of tissue found in various regions around the prosthesis varied little. Further, the numbers of particles in tissue samples shown to have a chronic foreign‐body reaction was > 1 × 109 particles/gram. © 2003 Wiley Periodicals, Inc. J Biomed Mater Res 65A: 95–108, 2003
Diabetes, oxidative stress, and antioxidants: A review
Authors: Maritim A. C. , Sanders R. A. , Watkins J. B.
Published: March 2003

Increasing evidence in both experimental and clinical studies suggests that oxidative stress plays a major role in the pathogenesis of both types of diabetes mellitus. Free radicals are formed disproportionately in diabetes by glucose oxidation, nonenzymatic glycation of proteins, and the subsequent oxidative degradation of glycated proteins. Abnormally high levels of free radicals and the simultaneous decline of antioxidant defense mechanisms can lead to damage of cellular organelles and enzymes, increased lipid peroxidation, and development of insulin resistance. These consequences of oxidative stress can promote the development of complications of diabetes mellitus. Changes in oxidative stress biomarkers, including superoxide dismutase, catalase, glutathione reductase, glutathione peroxidase, glutathione levels, vitamins, lipid peroxidation, nitrite concentration, nonenzymatic glycosylated proteins, and hyperglycemia in diabetes, and their consequences, are discussed in this review. In vivo studies of the effects of various conventional and alternative drugs on these biomarkers are surveyed. There is a need to continue to explore the relationship between free radicals, diabetes, and its complications, and to elucidate the mechanisms by which increased oxidative stress accelerates the development of diabetic complications, in an effort to expand treatment options. © 2003 Wiley Periodicals, Inc. J Biochem Mol Toxicol 17:24–38, 2003; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.10058
Why some organizations go “beyond budgeting”
Authors: Player Steve
Published: March 2003

The vision of going “beyond budgeting” is to transform the performance potential of an
organization. This is done by freeing the organization from the performance trap of annual budgets and, at the
same time, releasing the full power of people in the front lines. It's a radical approach, but it's
been proven, and the potential benefits are both huge and enduring. © 2003 Wiley Periodicals,
Inc.
Identification of reciprocally regulated gene modules in regenerating dorsal root ganglion neurons and activated peripheral or central nervous system glia
Authors: Cameron Adrian A. , Vansant Gordon , Wu Wen , Carlo Dennis J. , ILL Charles R.
Published: March 2003

Differential gene expression in the rat after injury of dorsal root ganglion neurons in vivo, and simulation injury of Schwann cells and oligodendrocytes in vitro was analyzed using high‐density cDNA microarrays. The analyses were carried out to study the genetic basis of peripheral nerve regeneration, and to compare gene regulation in glia of the central (oligodendrocyte) and peripheral (Schwann cell) nervous systems. The genes showing significant differential regulation in the three study groups represented all aspects of cellular metabolism. However, two unexpected observations were made. Firstly, a number of identical genes were differentially regulated in activated Schwann cells, activated oligodendrocytes and regenerating DRG neurons. Specifically, a group of 113 out of 210 genes that were down‐regulated in Schwann cells upon lipopolysaccharide (LPS) treatment, were identical to genes up‐regulated in the injured, regenerating DRG. Furthermore, a group of 53 out of 71 genes that were down‐regulated in interferon gamma (IFN‐γ)/LPS‐activated oligodendrocytes, were identical to genes up‐regulated in the DRG neurons. Finally, 22 genes were common to these three groups, i.e., down‐regulated in activated oligodendrocytes, down‐regulated in activated Schwann cells, and up‐regulated in regenerating DRG neurons. Secondly, a group of 16 cell‐cycle and proliferation‐related genes were up‐regulated in the DRG following sciatic nerve crush, despite the absence of cells undergoing mitosis in the DRG, or any significant presence of apoptosis‐related gene expression. Therefore, it appears that in these three cell types, large sets of genes are reciprocally regulated upon injury and/or activation. This suggests that the activation of the injury‐related gene expression program in cell derivatives of the neuroectoderm involves, in part, highly conserved genetic elements. J. Cell. Biochem. 88: 970–985, 2003. © 2003 Wiley‐Liss, Inc.
Mouse PSP94 expression is prostate tissue‐specific as demonstrated by a comparison of multiple antibodies against recombinant proteins
Authors: Thota Anuradha , Karajgikar Meera , Duan Wenming , Gabril Manal Y. , Chan Franky L. , Wong Yong‐Chuan , Sakai Hideki , Chin Joseph L. , Moussa Madeleine , Xuan Jim W.
Published: March 2003

Prostate tissue‐specific gene expression is crucial for driving potentially therapeutic genes to target specifically to the prostate. Prostate secretory protein of 94 amino acids (PSP94), also known as β‐MSP (microseminoprotein), is one of the three most abundant secretory proteins of the prostate gland, and is generally considered to be prostate tissue‐specific. We have previously demonstrated that the expression of the rat PSP94 gene is strictly prostate tissue‐specific by an antibody against a recombinant rat PSP94. In order to study prostate targeting utilizing the PSP94 gene in a mouse pre‐clinical experimental model, we need to establish antibodies against mouse PSP94 to confirm if it is prostate tissue‐specific as well. In this study, firstly we raised a polyclonal antibody against a recombinant glutathione‐S‐transferase‐ (GST‐) mouse mature form of PSP94. However, it showed very poor immunoreactivity against prostate tissue PSP94 as tested in Western blotting experiments. Neither antibodies against rat PSP94 nor mouse PSP94 showed significant cross‐reactivity. Thus a second antibody was established against a recombinant mouse mature PSP94 containing N‐terminal polyhistidines, and stronger immunoreactivity against mouse prostate tissue PSP94 was identified in Western blotting experiments. Both of these antibodies showed immunohistochemical reactivity, while the latter showed stronger reactivity in IHC when tested with different fixatives. By studying tissue distribution, we demonstrated that, as with rat PSP94, mouse PSP94 is strictly prostate tissue‐specific in experiments of both Western blotting and immunohistochemistry (IHC). This conclusion was also derived from a comparison among antibodies against human, rat, and mouse PSP94, showing very different immunoreactivities in Western blotting and IHC. Finally, a competitive assay between different species was performed. We demonstrated that antibodies against PSP94 from different species (human, primate, rodents) have poor cross‐reactivities. These observations also indicate that the PSP94 gene is a rapidly evolving gene in all species. Results from this study have led to the possibility of utilizing PSP94 as a targeting agent specifically to the prostate in a mouse experimental model. J. Cell. Biochem. 88: 999–1011, 2003. © 2003 Wiley‐Liss, Inc.
Modulation of TNF‐α expression in bone marrow macrophages: Involvement of vitamin D response element
Authors: Hakim Imad , Bar‐Shavit Zvi
Published: March 2003

The calcium‐regulating hormone, 1,25(OH)2D3, induces tumor necrosis factor‐α (TNF‐α) synthesis and release from bone marrow macrophages (BMMs). To investigate the mechanism of this regulation, we have examined the effects of 1,25(OH)2D3 on the cytokine message. 1,25(OH)2D3 increased TNF‐α mRNA abundance in a dose‐ and time‐dependent manner. The combined treatment of BMMs with LPS and 1,25(OH)2D3 resulted in a synergistic increase of TNF‐α. The steroid also increased the expression of CD14 (LPS receptor). Vitamin D receptors (VDRs) mediate 1,25(OH)2D3 genomic effects by forming homodimers or heterodimers with retinoic acid receptors (RARs) or retinoic X receptors (RXRs). The RXR ligand, 9‐cis retinoic acid (9cRA), reduced TNF‐α mRNA abundance in BMMs, but increased CD14 mRNA levels. 1,25(OH)2D3 or LPS did not affect TNF‐α transcript stability. 9cRA, however, caused TNF‐α mRNA destabilization. Next, we searched for potential vitamin D response elements (VDREs) in the promoter region (1.2 kb) of the TNF‐α gene, and identified six such sequences. Using electrophoresis mobility shift assay (EMSA) we identified one of those sequences (−1008 to −994) as a likely candidate to be a VDRE (tnfVDRE). The binding of tnfVDRE to BMM‐derived nuclear extract was increased following cell treatment with 1,25(OH)2D3. No induction was observed with 9cRA treatment, but the retinoid enhanced the activity of 1,25(OH)2D3 when added together. Previously characterized VDREs (mouse osteopontin and rat osteocalcin) competed effectively with tnfVDRE, demonstrating the nature of the TNF‐α‐derived sequence as a VDRE. We observed super‐shift and block‐shift of the complex in the presence of either anti‐VDR or anti‐RXR antibodies. Our data suggest that 1,25(OH)2D3 increases TNF‐α transcript abundance in BMMs via a transcriptional mechanism; 9cRA decreases TNF‐α mRNA by destabilizing the transcript, and possibly also by forming transcriptionally inactive complex with 1,25(OH)2D3 on the tnfVDRE. The receptor complex interacting with tnfVDRE found in the promoter of the cytokine gene is probably composed of VDR‐RXR heterodimer. J. Cell. Biochem. 88: 986–998, 2003. © 2003 Wiley‐Liss, Inc.
α4 integrin increases anoikis of human osteosarcoma cells
Authors: Marco Rex A. W. , Díaz‐Montero C. Marcela , Wygant James N. , Kleinerman Eugenie S. , McIntyre Bradley W.
Published: March 2003

Cell motility, growth, and proliferation are regulated by adhesion to the extracellular matrix. Detachment of adherent cells from extracellular matrix results in induction of apoptosis (“anoikis”). Transformed cells often show an anchorage‐independent growth that enables them to acquire a motile, invasive phenotype. This phenotype has been associated with the altered expression and function of the integrin family of transmembrane proteins that mediate cell adhesion to the extracellular matrix. Although α4 integrin is normally expressed on leukocyte subpopulations, a number of metastatic melanomas and sarcomas express it as well. In this study, we demonstrated the expression of α4 integrins on the human osteosarcoma cell line SAOS and on metastatic osteosarcoma lesions from the lung and pericardium. We further demonstrated that α4 integrin is coupled to the β1 subunit by biochemical analysis and by using a mAb directed against a combinatorial epitope unique to the α4β1 molecule. SAOS cells undergo anoikis when adherence is denied. Anoikis involved the activation of caspase 3 and the release of cytochrome c from mitochondria. Treatment of non‐adherent SAOS with an anti‐α4 mAb increased anoikis while anti‐β1 integrin mAbs did not alter anoikis, thus indicating a novel function for the α4 subunit in the control of cell death. Since integrins can control cell migration, proliferation, and apoptosis these results demonstrate a potential role for α4 integrin during multiple aspects of osteosarcoma metastasis. J. Cell. Biochem. 88: 1038–1047, 2003. © 2003 Wiley‐Liss, Inc.
Concerning the reaction between singlet nitrenium ions and water: A computational investigation on competitive reaction paths
Authors: Facchini Paola , Grandinetti Felice
Published: March 2003

The reaction between singlet nitrenium ions XNH+ (X = F and Cl) and H2O has been investigated by high‐level of theory ab initio calculations. The geometries of the involved intermediates, transition structures, and dissociation products have been optimized at the MP2(full)/6‐31G(d) level of theory, and accurate total energies have been obtained using the Gaussian‐3 (G3) procedure. The reaction commences by the exothermic formation of the FNHOH and ClNHOH intermediates, which are in turn able to undergo two distinct low‐energy reaction paths, namely, the isomerization to the N‐protonated isomers of the hydroxylamines FNHOH or ClNHOH, and the eventual extrusion of HF or HCl. The competitive or alternative occurrence of these two processes strictly depends on the nature of the substituent X. In the reaction between FNH+ and H2O, the energy gained in the formation of the complex FNHOH from the association between FNH+ and H2O, 52.1 kcal mol−1, is by far larger than the activation barrier for the loss of HF from FNHOH, computed as 24.9 kcal mol−1. In addition, the FNHOH intermediate requires 33.0 kcal mol−1 to overcome the barrier for the isomerization to FNH2OH+. Therefore, the reaction between FNH+ and H2O is expected to occur practically exclusively by HF elimination with formation of the HNOH+ ionic product. On the other hand, for the reaction between ClNH+ and H2O, it is not possible to get a definitive conclusion on the competitive or alternative occurrence of the two reaction paths. In fact, the transition structure involved in the elimination of HCl from ClNHOH is only 3.4 kcal mol−1 lower in energy than the transition structure for the isomerization of ClNHOH to ClNH2OH+. In addition, the absolute values of the energy barriers of these two processes, 24.2 and 27.6 kcal mol−1, respectively, are comparable with the energy gained in the formation of the complex ClNHOH from the association between ClNH+ and H2O, 24.0 kcal mol−.1 Therefore, the ClNH+ cation is predicted to react with water significantly slower than FNH+. © 2003 Wiley Periodicals, Inc. J Comput Chem 24: 547–564, 2003
Development of hardware accelerator for molecular dynamics simulations: A computation board that calculates nonbonded interactions in cooperation with fast multipole method
Authors: Amisaki Takashi , Toyoda Shinjiro , Miyagawa Hiroh , Kitamura Kunihiro
Published: March 2003

Evaluation of long‐range Coulombic interactions still represents a bottleneck in the molecular dynamics (MD) simulations of biological macromolecules. Despite the advent of sophisticated fast algorithms, such as the fast multipole method (FMM), accurate simulations still demand a great amount of computation time due to the accuracy/speed trade‐off inherently involved in these algorithms. Unless higher order multipole expansions, which are extremely expensive to evaluate, are employed, a large amount of the execution time is still spent in directly calculating particle–particle interactions within the nearby region of each particle. To reduce this execution time for pair interactions, we developed a computation unit (board), called MD‐Engine II, that calculates nonbonded pairwise interactions using a specially designed hardware. Four custom arithmetic‐processors and a processor for memory manipulation (“particle processor”) are mounted on the computation board. The arithmetic processors are responsible for calculation of the pair interactions. The particle processor plays a central role in realizing efficient cooperation with the FMM. The results of a series of 50‐ps MD simulations of a protein–water system (50,764 atoms) indicated that a more stringent setting of accuracy in FMM computation, compared with those previously reported, was required for accurate simulations over long time periods. Such a level of accuracy was efficiently achieved using the cooperative calculations of the FMM and MD‐Engine II. On an Alpha 21264 PC, the FMM computation at a moderate but tolerable level of accuracy was accelerated by a factor of 16.0 using three boards. At a high level of accuracy, the cooperative calculation achieved a 22.7‐fold acceleration over the corresponding conventional FMM calculation. In the cooperative calculations of the FMM and MD‐Engine II, it was possible to achieve more accurate computation at a comparable execution time by incorporating larger nearby regions. © 2003 Wiley Periodicals, Inc. J Comput Chem 24: 582–592, 2003
Sparse matrix multiplications for linear scaling electronic structure calculations in an atom‐centered basis set using multiatom blocks
Authors: Saravanan Chandra , Shao Yihan , Baer Roi , Ross Philip N. , Head–Gordon Martin
Published: March 2003

A sparse matrix multiplication scheme with multiatom blocks is reported, a tool that can be very useful for developing linear‐scaling methods with atom‐centered basis functions. Compared to conventional element‐by‐element sparse matrix multiplication schemes, efficiency is gained by the use of the highly optimized basic linear algebra subroutines (BLAS). However, some sparsity is lost in the multiatom blocking scheme because these matrix blocks will in general contain negligible elements. As a result, an optimal block size that minimizes the CPU time by balancing these two effects is recovered. In calculations on linear alkanes, polyglycines, estane polymers, and water clusters the optimal block size is found to be between 40 and 100 basis functions, where about 55–75% of the machine peak performance was achieved on an IBM RS6000 workstation. In these calculations, the blocked sparse matrix multiplications can be 10 times faster than a standard element‐by‐element sparse matrix package. © 2003 Wiley Periodicals, Inc. J Comput Chem 24: 618–622, 2003
New enzymatic assay for serum urea nitrogen using urea amidolyase
Authors: Kimura Shigeki , Iyama Shigeru , Yamaguchi Yoshihisa , Kanakura Yuzuru
Published: March 2003

We established an enzymatic assay for measurement of serum urea nitrogen using urea amidolyase (EC 3.5.1.45) from yeast species. The method is based on hydrolysis of urea by the enzyme. In this assay, we eliminated endogenous ammonium ion by use of glutamate dehydrogenase (EC 1.4.1.4). Then in the presence of urea amido‐lyase, ATP, bicarbonate, magnesium, and potassium ions, ammonium ion was produced proportionally to urea concentration in serum. The concentra‐tion of ammonium ion formed was determined by adding GLDH to produce NADP+ in the presence of 2‐oxoglutarate and NADPH. We then monitored the change of absorbance at 340 nm. The inhibitory effect of calcium ion on this assay was eliminated by adding glyco‐letherdiamine‐N, N, N′, N′‐tetraacetic acid to the reaction system. The with‐in‐assay coefficient of variations (CVs) of the present method were 1.80–3.76% (n = 10) at 2.8–19.0 mmol/L, respectively. The day‐to‐day CVs were 2.23–4.59%. Analytical recovery was 92–115%. The presence of ascorbic acid, bilirubin, hemoglobin, lipemic material, ammo‐nium ion, or calcium ion did not affect this assay system. The correlation be‐tween values obtained with the present method (y) and those by another enzy‐matic method (x) was 0.997 (y = 1.02x − 0.10 mmol/L, Sy/x = 0.841, n = 100), with a mean difference of –0.18 ± 0.86 mmol/L [(values by reference method – that of present method) ± SD] using the Bland‐Altman technique. J. Clin. Lab. Anal. 17:52–56, 2003. © 2003 Wiley‐Liss, Inc.
The Depression—Arkansas scale: A validation study of a new brief depression scale in an HMO
Authors: Walter Lawrence J. , Meresman Joel F. , Kramer Teresa L. , Evans Richard B.
Published: March 2003

Recent trends in mental‐health care have increased the need for practical depression instruments. The Depression—Arkansas (D‐ARK), a brief, economical, multipurpose instrument, has been validated for assessing major depressive disorder (MDD) and depressive‐symptom severity. Psychometric properties of the D‐ARK were compared with standard depression scales (Beck Depression Inventory and Geriatric Depression Scale) among 294 adult and 193 senior primary‐care patients, respectively, and 163 patients enrolled in cognitive–behavioral depression classes. The severity scale displayed adequate internal reliability (coefficient α = .81–.86), high correlation with the BDI‐2 (r = .78–.83) and GDS (r = .75), and similar factor structure to the BDI‐2. The D‐ARK was calibrated against the BDI‐2 and GDS, providing familiar severity category cutpoints with the new instrument. This study yields further data supporting the reliability, validity, and practical utility of the D‐ARK. © 2003 Wiley Periodicals, Inc. J Clin Psychol 59: 465–481, 2003.
Regulatory mechanisms of periodontal regeneration
Authors: Shimono Masaki , Ishikawa Tatsuya , Ishikawa Hiroki , Matsuzaki Hirokazu , Hashimoto Sadamitsu , Muramatsu Takashi , Shima Kaori , Matsuzaka Ken‐Ichi , Inoue Takashi
Published: March 2003

The periodontal ligament, located between the cementum and the alveolar bone, has a width ranging from 0.15 to 0.38 mm. Regeneration and homeostasis of the periodontal ligament are highly significant functions in relation to periodontal therapy, tooth transplantation or replantation, and orthodontic tooth movement. The purpose of this review is to discuss the regulatory mechanisms of regenerative and homeostatic functions in the periodontal ligament based on currently published studies and also on our own experimental data. We consider the capability of the ligament tissue to promote or to suppress calcification in connection with bone and cementum formation and the maintenance of the periodontal ligament space. Also discussed are the involvement of the periodontal ligament tissue in the regenerative ability, cell proliferation, growth and differentiation factors, extracellular matrix proteins, homeostatic phenomena, function of Malassez epithelial rests, tooth movement, or occlusal loading. Regulatory mechanisms for regeneration and homeostasis of the periodontal ligament are hypothetically proposed. Microsc. Res. Tech. 60:491–502, 2003. © 2003 Wiley‐Liss, Inc.
Help, risk and deceit: microentrepreneurs talk about microfinance
Authors: Eversole Robyn
Published: March 2003

Microenterprise lending has emerged as an important development strategy internationally. Its goal is to support micro and small businesses as motors for impoverished local economies and to build sustainable microfinance organizations to serve these businesses. This article explores the experiences of microentrepreneurs in Sucre, Bolivia to probe the relationships underlying the microenterprise lending process. This study illustrates how these ostensibly commercial transactions were grounded in complex assumptions about social development, external assistance and power. The article illustrates the divide between ‘developers’ and the ‘developed’ in their shared quest to ‘help businesses grow’, and concludes that building strong lending institutions does not automatically translate into broad‐based benefits for microentrepreneurs or their businesses. Copyright © 2003 John Wiley & Sons, Ltd.
Epidermal differentiation during ontogeny and after hatching in the snake Liasis fuscus (Pythonidae, Serpentes, Reptilia), with emphasis on the formation of the shedding complex
Authors: Alibardi L. , Thompson M.B.
Published: March 2003

Differentiation and localization of keratin in the epidermis during embryonic development and up to 3 months posthatching in the Australian water python, Liasis fuscus, was studied by ultrastructural and immunocytochemical methods. Scales arise from dome‐like folds in the skin that produce tightly imbricating scales. The dermis of these scales is completely differentiated before any epidermal differentiation begins, with a loose dermis made of mesenchymal cells beneath the differentiating outer scale surface. At this stage (33) the embryo is still unpigmented and two layers of suprabasal cells contain abundant glycogen. At Stage 34 (beginning of pigmentation) the first layers of cells beneath the bilayered periderm (presumptive clear and oberhautchen layers) have not yet formed a shedding complex, within which prehatching shedding takes place. At Stage 35 the shedding complex, consisting of the clear and oberhautchen layers, is discernible. The clear layer contains a fine fibrous network that faces the underlying oberhautchen, where the spinulae initially contain a core of fibrous material and small beta‐keratin packets. Differentiation continues at Stage 36 when the beta‐layer forms and beta‐keratin packets are deposited both on the fibrous core of the oberhautchen and within beta‐cells. Mesos cells are produced from the germinal layer but remain undifferentiated. At Stage 37, before hatching, the beta‐layer is compact, the mesos layer contains mesos granules, and cells of the alpha‐layer are present but are not yet keratinized. They are still only partially differentiated a few hours after hatching, when a new shedding complex is forming underneath. Using antibodies against chick scale beta‐keratin resolved at high magnification with immunofluorescent or immunogold conjugates, we offer the first molecular confirmation that in snakes only the oberhautchen component of the shedding complex and the underlying beta cells contain beta‐keratin. Initially, there is little immunoreactivity in the small beta‐packets of the oberhautchen, but it increases after fusion with the underlying cells to produce the syncytial beta layer. The beta‐keratin packets coalesce with the tonofilaments, including those attached to desmosomes, which rapidly disappear in both oberhautchen and beta‐cells as differentiation progresses. The labeling is low to absent in forming mesos‐cells beneath the beta‐layer. This study further supports the hypothesis that the shedding complex in lepidosaurian reptiles evolved after there was a segregation between alpha‐keratogenic cells from beta‐keratogenic cells during epidermal renewal. J. Morphol. 256:29–41, 2003. © 2003 Wiley‐Liss, Inc.
Autapomorphies of the endophragmal system in trichodactylid freshwater crabs (Crustacea: Decapoda: Eubrachyura)
Authors: Sternberg Richard v. , Cumberlidge Neil
Published: March 2003

An examination of the thoracic endophragmal system in trichodactylid freshwater crabs revealed several derived features that appear restricted to this family. Trichodactylids have a highly reduced sella turcica with a rim‐like overall configuration; as a consequence of sella turcica reduction the median septum is dorsoventrally diminished and exposed along its length. In addition, interosternite 7/8 is medially erased with the remnants restricted to the lateral regions of the sternum. The configurations of the sella turcica and interosternite 7/8 in trichodactylids underscore the monophyly of this family. The Dilocarcininae is characterized by having interosternites 5/6 through 7/8 restricted to the lateral sectors of the sternum, the lateral remnants of interosternite 4/5 forming prominent, rib‐like ridges oriented diagonally to the longitudinal axis of the crab, and the median septum forming with interosternite 4/5 a Y‐shaped pattern. These autapomorphies of the endophragmal system separate the trichodactylids from all other eubrachyurans and emphasize the morphological distinctness of the group. A description of the trichodactylid endophragmal junction plate is also provided. J. Morphol. 256:23–28, 2003. © 2003 Wiley‐Liss, Inc.
Anatomical features of Leiopelma embryos and larvae: Implications for anuran evolution
Authors: Bell Ben D. , Wassersug Richard J.
Published: March 2003

A controversial issue in anuran systematics is the relationship of Leiopelma to other anurans because recent phylogenetic constructions imply different relationships among the basal frogs. Of particular evolutionary interest is whether early development of Leiopelma resembles an ancestral salamander‐like larva, an anuran tadpole, or neither. In the 1950s, Neville G. Stephenson hypothesized that direct development is the primary mode of development in amphibians, based on the fact that Leiopelma spp. lack a free‐living (=feeding) larval stage. Although this hypothesis has not been generally accepted, it has not been formally refuted. We review Stephenson's work on Leiopelma and examine the anatomy of embryos/“larvae” of the four extant Leiopelma species for evidence of vestigial larval features that might refute the “direct‐developing ancestor” hypothesis. We describe internal oral features in early developmental stages of Leiopelma and compare Leiopelma with a closely related basal anuran, Ascaphus, to assess whether their early developmental stages share any derived features. In Leiopelma hochstetteri, embryos/larvae have open gill slits and some faint rugosities around one gill slit that may be vestiges of gill rakers or filters. They also have more intestinal loops, indicative of an elongated alimentary tract, at earlier rather than late embryonic/larval stages. Collectively, these features support the view that the ancestor of Leiopelma had a free‐swimming, free‐feeding, aquatic larva. The palatoquadrate of Leiopelma archeyi reorients approximately 40° from a more horizontal to a more vertical position through embryonic/“larval” development. This amount of cranial remodeling is intermediate between that seen in salamanders (17–27°) and that reported for Ascaphus (64°) and other basal frogs (71–78°) at metamorphosis. We found no internal oral features that Leiopelma shares specifically with Ascaphus. However, Leiopelma embryos have a ventrally positioned mouth and a downturned rostrum, characteristic of Ascaphus and other stream‐adapted tadpoles. J. Morphol. 256:160–170, 2003. © 2003 Wiley‐Liss, Inc.
Mutational patterns of paired blood and rectal biopsies in HIV‐infected patients on HAART
Authors: Monno Laura , Punzi Grazia , Scarabaggio Teresa , Saracino Annalisa , Brindicci Gaetano , Fiore José Ramon , Caputi Iambrenghi Onofrio , Di Stefano Mariantonietta , Pastore Giuseppe , Angarano Gioacchino
Published: March 2003

Blood and concurrent rectal biopsy samples of human immunodeficiency virus type 1 (HIV‐1)‐positive highly active antiretroviral therapy (HAART)‐treated patients were tested for genotypic resistance by direct sequencing of reverse transcriptase (RT) and protease (PR) regions to compare the patterns of resistance in these compartments. Fourteen subjects (five with undetectable plasma viral load (pVL) and nine persistently viremic) were studied. Four of five patients with undetectable pVL also had undetectable mucosal HIV RNA; sequence analyses from proviral DNA (PBMCs and rectal biopsy) were obtained with none or few resistance‐associated mutations and no alteration of susceptibility profile. All viremic patients, and one with negative pVL, had detectable levels of mucosal HIV RNA (1.93–4.21 log10 copies/mg); sequences of HIV RNA (plasma and/or rectal biopsy) were also obtained, and multiple mutations generally compatible with current/past medications were detected. Overall, 40 HIV‐1 PR and 42 RT sequences were analyzed, yielding a total of 42 PR and 47 RT sequence pairs (plasma/tissue‐RNA; plasma‐RNA/tissue‐DNA; PBMC/tissue‐DNA; tissue‐DNA/RNA; tissue‐RNA/PBMC‐DNA; PBMC‐DNA/plasma‐RNA), which almost always differed at the total amino acid level (median percentage discordance 8.08% in the PR, 4.8% in RT). The median percentage of resistance position discordance equaled 88.8% (IQR = 20–100) in the PR and 74.55% (IQR = 31.75–100%) in the RT pairs, respectively. Different resistance levels were detected by means of a computer‐assisted interpretation of mutational profiles. The results support the multiform evolution of HIV genotype in various body compartments and emphasize the participation of intestinal mucosa in HIV genotype selection. Samples from diverse tissues should be used for resistance evaluation to obtain a complete picture of drug resistance for antiretroviral‐treated patients. J. Med. Virol. 70: 1–9, 2003. © 2003 Wiley‐Liss, Inc.
Elevated levels of total and dengue virus‐specific immunoglobulin E in patients with varying disease severity
Authors: Koraka Penelopie , Murgue Bernadette , Deparis Xavier , Setiati Tatty E. , Suharti Catarina , van Gorp Eric C.M. , Hack C.E. , Osterhaus Albert D.M.E , Groen Jan
Published: March 2003

The kinetics of total and dengue virus‐specific immunoglobulin E (IgE) were studied in serial serum samples obtained from 168 patients, 41 of whom suffered from primary dengue virus infection and 127 suffered from secondary dengue virus infection. Seventy‐one patients were classified as dengue fever, 30 as dengue hemorrhagic fever, and 67 as dengue shock syndrome. A control group included single serum samples from patients with a herpes virus infection (n = 14), non‐dengue febrile patients (n = 10), and healthy blood donors (n = 10). Patients with dengue virus infection had higher levels of total and dengue virus‐specific IgE than non‐dengue patients (P < 0.05). Patients with secondary dengue virus infections had not significantly increased levels of both total and dengue virus‐specific IgE in the acute phase of disease compared to patients with primary dengue virus infections. Dengue virus‐specific IgE was significantly higher in dengue hemorrhagic fever and/or dengue shock syndrome patients compared to dengue fever and non‐dengue patients (P < 0.05). In conclusion, this study showed elevated total and dengue virus‐specific IgE serum antibody levels in the acute stage of disease. Therefore, measurement of both total and dengue virus‐specific IgE serum antibodies can be used as an additional prognostic marker in the development of severe complications in dengue virus infections. In addition, the presence and increase of dengue virus‐specific IgE serum antibodies in patients with dengue virus infections is suggestive of the pathogenetic role that IgE may play in the hemostatic disorders observed in dengue hemorrhagic fever and dengue shock syndrome. J. Med. Virol. 70:91–98, 2003. © 2003 Wiley‐Liss, Inc.
Recombinant murine L20B cell line supports multiplication of group A coxsackieviruses
Authors: Nadkarni Sushma S. , Deshpande Jagadish M.
Published: March 2003

Sensitive, reliable, and rapid methods of virus culture are essential for wild poliovirus isolation and identification from stool specimens collected from cases of acute flaccid paralysis. Recently, recombinant murine cell lines expressing human poliovirus receptor (CD155) on the cell surface have become available. These cells are sensitive selectively to poliovirus because the poliovirus receptor is not used by other enteroviruses. In early field studies non‐polio enteroviruses were not isolated from stool samples of cases of acute flaccid paralysis using L20B cells. For the past 3 years, L20B cells were used extensively in our laboratory for virus culture. The objective of the present study was to identify non‐polio enteroviruses causing cytopathic changes in L20B cells. Stool specimens collected from 1,153 cases of acute flaccid paralysis and 2,670 apparently healthy children were tested for enteroviruses using RD and L20B cell lines. A small number of viruses other than poliovirus causing cytopathic effect in L20B cells were isolated. Such isolates detected in other polio network laboratories in India were also included. The virus isolates were typed using partial VP1 nucleotide sequence analysis and virus neutralization tests. Of the 111 viruses studied, 8 were non‐enteroviruses. Among the 103 non‐polio enteroviruses, 73 were identified as group A coxsackieviruses. Of the 30 isolates that could not be characterized, 1 remained unidentified even by sequence analysis and 29 did not reach high titers in L20B as well as RD cells. In conclusion, coxsackie A viruses multiply in L20B cells causing cytopathic effect. Coxsackievirus A8 and coxsackievirus A10 were predominant among the eight coxsackie A virus types so far identified. J. Med. Virol. 70:81–85, 2003. © 2003 Wiley‐Liss, Inc.
Inhibition of enterovirus 71‐induced apoptosis by allophycocyanin isolated from a blue‐green alga spirulina platensis
Authors: Shih Shin‐Ru , Tsai Kun‐Nan , Li Yi‐Shuane , Chueh Chuang‐Chun , Chan Err‐Cheng
Published: March 2003

Enterovirus 71 infection causes significant morbidity and mortality in children, yet there is no effective treatment. In this study, a protein‐bound pigment, allophycocyanin purified from blue‐green algae is first reported to exhibit anti‐enterovirus 71 activity. Allophycocyanin neutralized the enterovirus 71‐induced cytopathic effect in both human rhabdomyosarcoma cells and African green monkey kidney cells. The 50% inhibitory concentration of allophycocyanin for neutralizing the enterovirus 71‐induced cytopathic effect was approximately 0.045 ± 0.012 μM in green monkey kidney cells. The cytotoxic concentrations of allophycocyanin for rhabdomyosarcoma cells and African green monkey kidney cells were 1.653 ± 0.003 μM and 1.521 ± 0.012 μM, respectively. A plaque reduction assay showed that the concentrations of allophycocyanin for reducing plaque formation by 50% were approximately 0.056 ± 0.007 μM and 0.101 ± 0.032 μM, when allophycocyanin were added at the state of viral adsorption and post‐adsorption, respectively. Antiviral activity was more efficient in cultures treated with allophycocyanin before viral infection compared with that in the cultures treated after infection. Allophycocyanin was also able to delay viral RNA synthesis in the infected cells and to abate the apoptotic process in enterovirus 71‐infected rhabdomyosarcoma cells with evidence of characteristic DNA fragmentation, decreasing membrane damage and declining cell sub‐G1 phase. It is concluded that allophycocyanin possesses antiviral activity and has a potential for development as an anti‐enterovirus 71 agent. J. Med. Virol. 70:119–125, 2003. © 2003 Wiley‐Liss, Inc.
Introduction of HIV type 1 non‐B subtypes into Eastern Andalusia through immigration
Authors: Alvarez Marta , García Federico , Martínez Nora Mariela , García Fernando , Bernal Carmen , Vela Carmen Maroto , Angulo Gonzalo Piédrola , Quero José Hernández
Published: March 2003

A study of the distribution of HIV‐1 subtypes in the native and immigrant populations of Eastern Andalusia (Southern Spain) was conducted to determine any changes between 1983 and 2001 and to identify antiretroviral resistance mutations in non‐B subtype strains among the immigrant population. The study included 111 native patients from Eastern Andalusia: 94 infected with HIV before 1996 and 17 infected since 1996. A parallel study was conducted on 26 HIV‐positive immigrants from Africa. Subtyping was done with the heteroduplex mobility assay. Resistance mutations were determined by line probe assay. A total of 137 patients were studied: 9.2% had subtype A (n = 12), 80.8% subtype B (n = 105), and 1.5% subtype C (n = 2). Among the Eastern Andalusia population infected before 1996, 10.9% had non‐B subtypes, compared with 23.5% of those infected after that year. The greatest percentage of non‐B subtypes (52.4%) was found among the immigrant population. Resistance mutation K70R was detected in one of the six immigrants with non‐B subtype and M41L in another. There has been a slight increase in the diversity of HIV‐1 subtypes in Eastern Andalusia over the past few years, possibly influenced by non‐B subtypes introduced by immigrants from sub‐Saharan Africa. J. Med. Virol. 70: 10–13, 2003. © 2003 Wiley‐Liss, Inc.
Epstein‐barr virus infection of rat lymphocytes expressing human CD21 results in restricted latent viral gene expression and not in immunoblastic transformation
Authors: Yang Lixin , Ikeda Hitoshi , Lai Yurong , Yoshiki Takashi , Takada Kenzo
Published: March 2003

Transgenic rats expressing human CD21 gene (hCD21) driven by the mouse immunoglobulin enhancer were generated. hCD21 was expressed in lymphoid tissues, especially in the spleen and in the brain. Flow cytometric analysis indicated that about 20% of spleen cells, most having a B‐lymphocyte marker, expressed hCD21. After Epstein‐Barr virus (EBV) infection of spleen cells, EBV‐determined nuclear antigen (EBNA) was first detected on Day 4 and reached a maximum of 0.3% on Day 5, but the infection was abortive and was not followed by blastogenesis, cellular DNA synthesis or proliferation. Reverse transcription‐polymerase chain reaction (RT‐PCR) analyses demonstrated that EBV‐infected spleen cells expressed EBNA1 and EBV‐encoded small RNA (EBER), but not other latent EBV products. EBNA promoter analysis by RT‐PCR indicated that the Q promoter was active, whereas C and W promoters were not active. The present findings indicate that human and rat lymphocytes respond to EBV infection differently in vitro. J. Med. Virol. 70:126‐130, 2003. © 2003 Wiley‐Liss, Inc.
GB Virus C genotype determination in GB Virus‐C/HIV co‐infected individuals
Authors: Muerhoff A. Scott , Tillmann Hans L. , Manns Michael P. , Dawson George J. , Desai Suresh M.
Published: March 2003

Several recent studies have indicated that patients infected with human immunodeficiency virus (HIV) exhibit a beneficial effect of co‐infection with GB virus C (GBV‐C). The benefit is demonstrated by slower progression to acquired immunodeficiency syndrome (AIDS) and prolonged survival time after the development of AIDS. In some but not all studies, a significant association between GBV‐C/HIV co‐infection and increased CD4+ cell counts has been reported. To understand further the possible role that GBV‐C might play in the reduced morbidity and mortality among HIV‐infected patients, we sought to examine the presence of different GBV‐C genotypes in a cohort of co‐infected patients. PCR products derived from the 5′‐untranslated region (5′‐UTR) and the second envelope gene (E2) were sequenced directly and genotyped by phylogenetic analysis. While 5′‐UTR analysis delineated the major type, analysis of the complete E2 gene was required for identification of group 2 subtypes, designated 2a and 2b. Among 35 patients tested, GBV‐C genotype was determined for 33: two patients were infected with genotype 1, 12 with type 2a, and 19 with type 2b. Clinical data were available for 25 genotyped patients: one infected with genotype 1, nine with genotype 2a, and 15 with type 2b. CD4 cell counts tended to be lower in patients infected with genotype 2a compared with those with genotype 2b (310 ± 136 vs 430 ± 199, P = 0.054). Additional studies with larger cohorts from separate geographical regions are needed to determine whether a particular GBV‐C genotype is associated with reduced morbidity or mortality among HIV co‐infected patients. J. Med. Virol. 70:141–149, 2003. © 2003 Wiley‐Liss, Inc.
Time‐domain simulation of the full hydrodynamic model
Authors: Aste Andreas , Vahldieck Rüdiger
Published: March 2003

A simple upwind discretization of the highly coupled non‐linear differential equations which define the hydrodynamic model for semiconductors is given in full detail. The hydrodynamic model is able to describe inertia effects which play an increasing role in different fields of opto‐ and microelectronics. A silicon n+−n−n+‐structure is simulated, using the energy‐balance model and the full hydrodynamic model. Results for stationary cases are then compared, and it is pointed out where the energy‐balance model, which is implemented in most of today's commercial semiconductor device simulators, fails to describe accurately the electron dynamics. Additionally, a GaAs n+−n−n+‐structure is simulated in time domain in order to illustrate the importance of inertia effects at high frequencies in modern submicron devices. Copyright © 2003 John Wiley & Sons, Ltd.
Hydroxyl radical formation is greater in striatal core than in penumbra in a rat model of ischemic stroke
Authors: Liu Shimin , Liu Miao , Peterson Steven , Miyake Minoru , Vallyathan Val , Liu Ke Jian
Published: March 2003

Although hydroxyl radical (•OH) formation has been implicated in the pathophysiological changes of ischemic stroke, •OH production in the core and penumbra regions is not clear. It is extremely important to distinguish penumbra from ischemic core in focal cerebral ischemia studies, because the penumbra contains viable tissue, which can be salvaged by appropriate treatment. This study evaluated •OH production in both core and penumbra regions of ischemic striatum during ischemia and reperfusion. Microdialysis probes were placed in striatal tissue of rats subjected to the middle cerebral artery occlusion model of ischemic stroke. The •OH‐trapping agent 4‐hydroxybenzoic acid (4‐HBA) was administered by both i.v. and probe infusion. Dialysate levels of the 4‐HBA oxidation products, 3,4‐dihydroxybenzoic acid (3,4‐DHBA), were determined by HPLC‐ECD. After microdialysis probe delivery of 4‐HBA, •OH production was significantly increased in the striatal core during both ischemia and reperfusion. Penumbra •OH production increased only during reperfusion. Alterations of 3,4‐DHBA concentration in dialysate following i.v. 4‐HBA administration were likely related to alterations in tissue blood flow. The findings were confirmed by a greater oxidation of dihydroethidium in the ischemic core than in the penumbra as determined by fluorescent microscopy. The findings of •OH production in ischemic striatum are the opposite of those reported for ischemic cortex and suggest critical regional variations in •OH production that may have significant clinical implications in the treatment of ischemic stroke. © 2003 Wiley‐Liss, Inc.
Neurotoxicity from glutathione depletion is dependent on extracellular trace copper
Authors: White Anthony R. , Cappai Roberto
Published: March 2003

Glutathione (GSH) is an important antioxidant, and its depletion in neurons has been implicated in several neurodegenerative disorders. Aberrant copper metabolism is also implicated in neurodegeneration and may result in the generation of toxic free radicals. However, little is known about the relationship between GSH depletion and copper homeostasis. In the present study, we examined the role of extracellular trace biometals in neuronal cell death induced by GSH depletion. Treatment of primary cortical neurons with buthionine sulfoximine (BSO), an inhibitor of GSH synthesis, induced a rapid loss of intracellular GSH, leading to decreased neuronal cell viability. Neuronal cell death induced by GSH depletion was dependent on trace levels of extracellular copper in the culture medium (1.6 μM). Neurons were protected against GSH depletion‐mediated toxicity when cultured in Chelex 100‐treated medium containing tenfold less copper (0.16 μM) than normal medium. The addition of copper, but not iron or zinc, to Chelex 100‐treated medium restored the neurotoxicity induced by GSH depletion. Moreover, BSO toxicity in normal medium was inhibited by copper chelators. The neurotoxic effects of copper in GSH‐depleted neurons involved generation of copper(I) and subsequent free radical‐mediated oxidative stress. These studies demonstrate a critical role for extracellular trace copper in neuronal cell death caused by GSH depletion and may have important implications for the understanding of toxic processes in neurodegenerative diseases. © 2003 Wiley‐Liss, Inc.