bluuebunny/crossref_metadata_2025 huggingface dataset:
Multifunctional anti‐angiogenic activity of the cyclic peroxide ano‐2 with antitumor activity
Authors: Arakawa Kazuhito , Endo Yoshio , Kimura Masashi , Yoshida Takanori , Kitaoka Takeshi , Inakazu Tomoyasu , Nonami Yuji , Abe Manabu , Masuyama Araki , Nojima Masatomo , Sasaki Takuma
Published: October 2002

Our focus was to develop an anti‐angiogenic drug possessing the inhibitory activity of urokinase‐type plasminogen activator (u‐PA) production. During preliminary screening, the effects of 13 ozonides on the inhibition of u‐PA production in human fibrosarcoma HT‐1080 cells and on the inhibition of angiogenesis on chicken embryonic chorioallantoic membranes were determined. Of the ozonides tested, 9 inhibited in vitro u‐PA production of HT‐1080 cells and 7 of these 9 exhibited strong anti‐angiogenic activity. Interestingly, 6 of the 13 ozonides also inhibited cathepsin B activity. 1‐Phenyl‐1, 4‐epoxy‐1H,4H‐naphtho[1,8‐de][1, 2]dioxepin (ANO‐2) potently inhibited cathepsin B (IC50 = 0.47 μM) as well as u‐PA production. Consequently, ANO‐2 was selected for further study. ANO‐2 inhibited tube formation by human umbilical vein endothelial cells cultured on Matrigel while exhibiting no cytotoxicity. Additionally, in vivo administration of ANO‐2 inhibited angiogenesis induced by mouse Sarcoma‐180 cells tested using the mouse dorsal air sac assay. Moreover, ANO‐2 also suppressed primary tumor growth and reduced the number of pulmonary metastases caused by Lewis lung carcinoma cells in mice. These in vitro and in vivo activities indicate that ANO‐2 has considerable potential as a new and potent anti‐angiogenic drug that inhibits both u‐PA production and enzymatic activity of cathepsins, indicating that ANO‐2 may be a multifunctional inhibitor of angiogenesis. © 2002 Wiley‐Liss, Inc.
Sustainable use of groundwater for irrigation: a numerical analysis of the subsoil water fluxes
Authors: Ahmad Mobin‐ud‐Din , Bastiaanssen W. G. M. , Feddes R. A.
Published: October 2002

The food‐producing regions of the world increasingly rely on irrigation from groundwater resources. Further increases of groundwater use can adversely affect the sustainability of irrigated agriculture and put food security at risk. Sustainability of irrigation at field scale with groundwater is obtained if groundwater recharge is in equilibrium with tubewell extractions and capillary rise. Traditional information on phreatic surface behaviour does not explain the processes causing a phreatic surface to decline or incline. In this study, the physically based numerical model Soil–Water–Atmosphere–Plant (SWAP) was applied to compute soil moisture content and vertical soil water fluxes in the unsaturated zone for the cotton–wheat and rice–wheat cropping system of Punjab, Pakistan. SWAP has been calibrated and verified with in situ measurements of soil moisture content and evapotranspiration fluxes measured by means of the Bowen ratio surface energy balance technique. Accurate data of the soil hydraulic properties are critical for the calibration of the soil moisture distribution. With knowledge of the van Genuchten–Mualem parameters available, SWAP could be applied to assess recharge and capillary rise for most field conditions, including basin irrigation. The results under Pakistani conditions show that deep percolation cannot always be estimated from root zone water balances. An annual recharge of 23.3 cm was computed for the cotton–wheat area. Sustainability of irrigation with groundwater is obtained if a reduction in irrigation with groundwater by 36% is obtained. An annual recharge of 38.9 cm is estimated in rice–wheat systems, and a reduction of 62% in groundwater extraction is required to reach sustainability of groundwater use at field scale. Such information cannot be obtained from classical phreatic surface fluctuation data, and unsaturated zone modelling therefore provides additional insights for groundwater policy making. Copyright © 2002 John Wiley & Sons, Ltd.
Bioactivity of three CaO–P2O5–SiO2 sol‐gel glasses
Authors: Salinas A. J. , Martin A. I. , Vallet‐Regí M.
Published: October 2002

Three gel glasses containing 25 mol % of CaO and SiO2 + P2O5 contents (in mol %): 75 + 0 (S75); 72.5 + 2.5 (S72.5P2.5); and 70 + 5 (S70P5), respectively, were obtained, characterized, and studied when soaked in a simulated body fluid (SBF). The influence of composition in both textural properties (surface area and porosity) and in vitro behavior of glasses was studied. In as prepared S72.5P2.5 and S70P5 glasses, crystalline phosphate nuclei were detected through XRD and FTIR. In addition, N2 adsorption and Hg porosimetry measurements showed that the surface area increased, whereas the pore volume and the pore diameter decreased as P2O5 in glasses increased. These variations were explained on the basis of the withdrawal of calcium from the glass silica network, due to the calcium‐phosphorous bonding. In vitro studies showed that the three compositions were bioactive, because an apatite layer was formed after soaking in SBF. S75 presented the highest initial reactivity but the lowest crystallization rate of the apatite‐like phase. For S72.5P2.5, and S70P5 the amorphous calcium phosphate formation was slower than for S75, but the crystallization of apatite was observed after shorter periods in SBF. Furthermore, after 7 days of soaking, the layer thickness decreased as P2O5 in glasses increased. © 2002 Wiley Periodicals, Inc. J Biomed Mater Res 61: 524–532, 2002
Mononuclear cell collection in patients undergoing extra‐corporeal photo‐chemotherapy for acute and chronic graft‐vs.‐host‐disease (GvHD): Comparison between COBE Spectra version 4.7 and 6.0 (AutoPBSC)
Authors: Perseghin Paolo , Dassi Maria , Balduzzi Adriana , Rovelli Attilio , Bonanomi Sonia , Uderzo Cornelio
Published: October 2002

A constant improvement in the performance of blood cell separators has been observed in recent years, allowing better yields in peripheral blood stem cell collection (PBSC) either from healthy donors or for autologous purposes. Nevertheless, to our knowledge, no reports on the efficiency of mononuclear cell (MNC) collection in patients undergoing extra‐corporeal photochemotherapy (ECP) for graft‐vs.‐host‐disease (GvHD) have been published. We retrospectively investigated the efficiency of 167 MNC collections performed consecutively in 12 patients between January 1999 and June 2001 by means of the COBE Spectra version 4.7 (V 4.7) or version 6.0 (V 6.0), for 109 and 58 procedures, respectively. MNC fractional extraction (FE) was higher in the V 6.0 group compared to the V 4.7 group : 0.59 ± 0.21 vs. 0.51 ± 0.22 (P < 0.05). However, platelet contamination was lower in the products obtained with V.6.0 compared to those obtained with V.4.7: 740 ( 630 × 103/(L vs. 2,073 ( 1,429 × 103/(L (P < 0.05). Only two patients with acute GvHD, both from V 4.7 group, required post‐ECP platelet transfusion. The recently released version 6.0 allowed a satisfactory MNC yield with minimal platelet contamination in patients scheduled to undergo ECP for acute or chronic GvHD. J. Clin. Apheresis 17:65–71, 2002. © 2002 Wiley‐Liss, Inc.
Retroviral mediated expression of the human myeloid nuclear antigen in a null cell line upregulates Dlk1 expression
Authors: Doggett Kevin L. , Briggs Judith A. , Linton MacRae F. , Fazio Sergio , Head David R. , Xie Jingping , Hashimoto Yuko , Laborda Jorge , Briggs Robert C.
Published: October 2002

The human myeloid nuclear differentiation antigen (MNDA) is a hematopoietic cell specific nuclear protein. MNDA and other related gene products interact with and alter the activity of a large number of proteins involved in regulating specific gene transcription. MNDA and related genes exhibit expression characteristics, which suggest functions unique to specific lineages of cells, in addition to mediating the effects of interferons. Cells of the human K562 myeloid line do not express MNDA and are relatively immature compared to lines that express MNDA (HL‐60, U937, and THP1). The hypothesis that MNDA influences the expression of specific genes was tested by creating MNDA expressing K562 cells using stable retroviral mediated gene transfer followed by evaluation of transcription profiles. Two macroarrays containing a total of 2,350 cDNAs of known genes showed a specific up‐regulation of Dlk1 expression in MNDA expressing K562 cell clones. Real time quantitative RT‐PCR analysis confirmed an average of over 3‐ and 7‐fold upregulation of Dlk1 in two clones of MNDA expressing K562 cells. The effects on Dlk1 were also confirmed by Northern blotting. Dlk1 is essential for normal hematopoiesis and abnormal expression is a proposed marker of myelodysplastic syndrome. Additional screening of transcription profiles after induced erythroid and megakaryoblastic differentiation showed no additional gene transcripts altered by the presence of MNDA. These results indicate that MNDA alters expression of a gene essential for normal hematopoiesis. J. Cell. Biochem. 86: 56–66, 2002. © 2002 Wiley‐Liss, Inc.
Purification and characterization of protein tyrosine phosphatase PTP‐MEG2
Authors: Qi Ying , Zhao Runxiang , Cao Hongxi , Sui Xingwei , Krantz Sanford B. , Zhao Z. Joe
Published: October 2002

PTP‐MEG2 is an intracellular protein tyrosine phosphatase with a putative lipid‐binding domain at the N‐terminus. The present study reports expression, purification, and characterization of the full‐length form of the enzyme plus a truncated form containing the catalytic domain alone. Full‐length PTP‐MEG2 was expressed with an adenovirus system and purified from cytosolic extracts of human 293 cells infected with the recombinant adenovirus. The purification scheme included chromatographic separation of cytosolic extracts on fast flow Q‐Sepharose, heparin‐agarose, l‐histidyldiazobenzylphosphonic acid agarose, and hydroxylapatite. The enrichment of PTP‐MEG2 from the cytosol was about 120‐fold. The truncated form of PTP‐MEG2 was expressed in E. coli cells as a non‐fusion protein and purified by using a chromatographic procedure similar to that used for the full‐length enzyme. The purified full‐length and truncated enzymes showed single polypeptide bands on SDS–polyacrylamide gel electrophoresis under reducing conditions and behaved as monomers on gel exclusion chromatography. With para‐nitrophenylphosphate and phosphotyrosine as substrates, both forms of the enzyme exhibited classical Michaelis‐Menten kinetics. Their responses to pH, ionic strength, metal ions, and protein phosphatase inhibitors are similar to those observed with other characterized tyrosine phosphatases. Compared with full‐length PTP‐MEG2, the truncated ΔPTP‐MEG2 displayed significantly higher Vmax and lower Km values, suggesting that the N‐terminal putative lipid‐binding domain may have an inhibitory role. The full‐length and truncated forms of PTP‐MEG2 were also expressed as GST fusion proteins in E. coli cells and purified to near homogeneity through affinity columns. However, the specific phosphatase activities of the GST fusion proteins were 10–25‐fold below those obtained with the correspondent non‐fusion proteins. J. Cell. Biochem. 86: 79–89, 2002. © 2002 Wiley‐Liss, Inc.
Effects of bone morphogenetic protein‐7 stimulation on osteoblasts cultured on different biomaterials
Authors: Açil Yahya , Springer Ingo N.G. , Broek Vanessa , Terheyden Hendrik , Jepsen Søren
Published: October 2002

The objective of the present study was to investigate the effects of an in vitro stimulation of human osteoblasts by recombinant human bone morphogenetic protein‐7 (rhBMP‐7) on the collagen types and the quantity of the collagen cross‐links synthesized in a three‐dimensional culture on various biomaterials for bone replacement. Trabecular bone chips were harvested from human iliac crests, and cell cultures were established at standard conditions. One hundred and fifty nanograms per milliliter of rhBMP‐7 was added. For the second passage a cell scraper was used to bring the cells into suspension, and 100 μl osteoblasts (at a density of 3.3 × 105) were transferred onto nine blocks of either Bio‐Oss®, Tutoplast®, or PepGen p‐15™. Blocks incubated with cells that were not treated with rhBMP‐7 served as controls. Cell colonization of the biomaterials was observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) after a period of 2, 4, and 6 weeks. Throughout the experiment medium, supernatants were collected and collagen was characterized by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). Finally, the collagen cross‐link residues hydroxylysylpyridinoline (HP) and lysylpyridinoline (LP) were quantified by HPLC. Within 4 weeks the cells became confluent on all of the studied biomaterials. All samples synthesized bone specific LP and collagen type I. However, in rhBMP‐7‐stimulated samples, the amount of HP and LP found was increased by 45% compared to non‐stimulated samples. Cell proliferation and collagen synthesis was similar on the different biomaterials, but was consistently reduced in specimen not stimulated with rhBMP‐7. In vitro stimulation of osteoblasts on Bio‐Oss, Tutoplast, or PepGen p‐15 with rhBMP‐7 and subsequent transplantation of the constructs might lead to an enhanced osseointegration of the biomaterials in vivo. J. Cell. Biochem. 86: 90–98, 2002. © 2002 Wiley‐Liss, Inc.
Characterization of the nuclear matrix proteins in a transgenic mouse model for prostate cancer
Authors: Leman Eddy S. , Arlotti Julie A. , Dhir Rajiv , Greenberg Norman , Getzenberg Robert H.
Published: October 2002

The nuclear matrix (NM) contains a number of proteins that have been found to be associated with transformation. We have previously identified changes in the NM associated with prostate cancer. In this study, we examine the molecular changes that are associated with prostate cancer development in transgenic adenocarcinoma of mouse prostate (TRAMP) model by studying the differences in the NM proteins (NMPs). We collected prostates from the TRAMP males at six critical time points: 6 weeks (puberty), 11 and 19 weeks (development of mild hyperplasia), 25 weeks (development of severe hyperplasia), 31 and 37 weeks (development of neoplasia). The nuclear matrices from the prostates collected at these time points were then isolated and the NMPs were characterized by high‐resolution two‐dimensional gel electrophoresis. We found three NMPs (E1A, E1B, and E1C) that were present in the 6‐week‐old prostate and two NMPs (E2A and E2B) that were present in the 11‐week‐old prostate. These NMPs were absent in the 31‐ and 37‐week‐old prostate. We also found five NMPs (E3A–E3E) that were present in the 31‐week‐old prostate, but absent in the earlier time points. In addition, three NMPs (Le1, Le2, Le3) were present at higher expression in the 6‐, 11‐, 19‐, and 25‐weeks old TRAMP prostates, but they were expressed lower during the development of neoplasia at 31‐ and 37‐weeks old. Identification of these NMPs permits the development of novel markers that can characterize various stages of prostate cancer development as well as potentially therapeutic targets. J. Cell. Biochem. 86: 203–212, 2002. © 2002 Wiley‐Liss, Inc.
Crystal structure predictions using five space groups with two independent molecules. The case of small organic acids
Authors: Van Eijck Bouke P.
Published: October 2002

Crystal structure generations with two independent molecules have been performed for a series of carboxylic acids, using a slightly modified version of the OPLS force field. It was found that in this way the experimental structures with one independent molecule were produced as special cases, except for the molecules with four or more internal degrees of freedom. This work shows that a search with two independent molecules in only five space groups, although costly in computer power, can automatically also find structures with one independent molecule in many supergroups. Considering the observed abundances of structural classes, such a search should cover more than 95% of the possible homomolecular crystal structures. © 2002 Wiley Periodicals, Inc. J Comput Chem 4: 456–462, 2002; DOI 10.1002/jcc.10042
Mechanistic aspects of the dehydration and dehydrohalogenation of halo‐hydroxyformaldoxime conformers. A quantum chemical model study
Authors: Tsipis Athanassios C. , Tsipis Constantinos A.
Published: October 2002

A detailed exploration of the configurational and conformational space of chloro‐ and bromo‐hydroxyformaldoximes,Xhfaox (X = Cl, Br) has been carried out with the aid of the B3LYP level of density functional theory, using the 6‐31G(d,p) basis set. The most stable configuration in each series of theClhfaoxandBrhfaoxconformers corresponds to theZ‐s‐cis, s‐transconfiguration, while the highest energyZ‐(s‐trans, s‐cis)conformers were found at 7.0(7.6) and 6.0(6.6) kcal mol−1, respectively, at the B3LYP(QCISD(T)) levels of theory. Saddle points were also located on the PES of theClhfaoxandBrhfaoxcompounds corresponding toZ‐(s‐cis, s‐cis)conformers at 13.8(14.9) and 13.6(14.6) kcal mol−1, respectively, at the B3LYP(QCISD(T)) levels. Upon dehydrationXhfaoxcould afford a number of isomeric CXNO species. The dehydration processes ofXhfaoxare predicted to be endothermic, the computed heats of reactions found in the range of 20.5 to 86.2 kcal mol−1and 15.9 to 100.4 kcal mol−1at the B3LYP and QCISD(T) levels, respectively. The reaction pathways for the addition of water to halo‐fulminates yielding the most stableXhfaoxconformers was predicted to be concerted with a single transition structure, but are asynchronous with activation barriers of 32.8 and 43.0 kcal mol−1for the chloro‐ and bromo‐derivatives, respectively. The PES governing the isomerization reactions of the CXNO isomers have also been calculated, and possible isomerization pathways have been delineated. Upon dehydrohalogenation theXhfaoxconformers yield hydroxy‐isocyanate or hydroxy‐fulminate, the former being more stable by 31.8(18.8) kcal mol−1at the B3LYP(QCISD(T)) levels of theory. The reaction pathways for the addition of HX to hydroxy‐isocyanate were predicted to be slightly exothermic, the heats of reactions being −3.2 and −5.5 kcal mol−1, respectively, and have to surmount high activation barriers of 39.7 and 35.0 kcal mol−1, respectively. Similarly, the addition of HX to hydroxy‐fulminate was predicted to be much more exothermic, the heats of reactions being −34.7 and −37.3 kcal mol−1, respectively, and have to surmount much lower activation barriers of only 10.5 and 7.5 kcal mol−1respectively, at the B3LYP level. Finally, calculated structures, relative stability, and bonding properties of all stationary points located on the PES of the systems and reactions studied are thoroughly discussed with respect to computed electronic properties. © 2002 Wiley Periodicals, Inc. J Comput Chem 23: 1266–1280, 2002
Orthogonal double covers by super‐extendable cycles
Authors: Bey Christian , Hartmann Sven , Leck Uwe , Leck Volker
Published: October 2002

An Orthogonal Double Cover (ODC) of the complete graph Kn by an almost‐hamiltonian cycle is a decomposition of 2Kn into cycles of length n−1 such that the intersection of any two of them is exactly one edge. We introduce a new class of such decompositions. If n is a prime, the special structure of such a decomposition allows to expand it to an ODC of Kn+1 by an almost‐hamiltonian cycle. This yields the existence of an ODC of Kp+1 by an almost‐hamiltonian cycle for primes p of order 3 mod 4 and its eventual existence for arbitrary primes p. © 2002 Wiley Periodicals, Inc. J Combin Designs 10: 283–293, 2002; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jcd.10011
A conjecture on small embeddings of partial Steiner triple systems
Authors: Bryant Darryn
Published: October 2002

A well‐known, and unresolved, conjecture states that every partial Steiner triple system of order u can be embedded in a Steiner triple system of order υ for all υ ≡ 1 or 3, (mod 6), υ ≥ 2u + 1. However, some partial Steiner triple systems of order u can be embedded in Steiner triple systems of order υ <2u + 1. A more general conjecture that considers these small embeddings is presented and verified for some cases. © 2002 Wiley Periodicals, Inc. J Combin Designs 10: 313–321, 2002; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jcd.10017
A longitudinal study of the effects of seizure symptoms on the Minnesota Multiphasic Personality Inventory—2 (MMPI‐2) clinical interpretation
Authors: Derry Paul A. , Harnadek Michael C.S. , McLachlan Richard S. , Sontrop Jennifer , Blume Warren T. , Girvin John P.
Published: October 2002

This study investigated whether the Minnesota Multiphasic Personality Inventory—2 (MMPI‐2) items reflecting seizure symptoms spuriously inflate MMPI‐2 scores in persons with epilepsy. In a repeated measures prospective design, patients with epilepsy completed the MMPI‐2 prior to epilepsy surgery and again two years later when they were either seizure‐free or not seizure‐free. Compared to baseline scores, only seizure‐free patients showed a decline in endorsement of seizure items, and only on one scale, Hypochondriasis (Hs). Clinical profiles were not meaningfully different when readministered when seizures were no longer present. Moreover, profiles generated by individuals with and without seizures were not significantly different. We conclude the MMPI‐2 is a valid test in an epilepsy population, as seizure content did not alter its clinical interpretation. In using the MMPI‐2 in patients with epilepsy, a modest correction on the Hs and Schizophrenia (Sc) scales is warranted in selected cases. In MMPI‐2 research, assessing both statistical and clinical significance is recommended. © 2002 Wiley Periodicals, Inc. J Clin Psychol 58: 817‐826.
Integrating diversity and fostering interdependence: Ecological lessons learned about refugee participation in multiethnic communities
Authors: Goodkind Jessica R. , Foster‐Fishman Pennie G.
Published: October 2002

Communities in the United States are becoming increasingly diverse, in part, because of the continual resettlement of refugees and immigrants from around the world. The promotion of refugees' well‐being and integration depends upon how our communities value diversity and provide opportunities for meaningful involvement. However, refugees often face challenges to such involvement. An ecological perspective suggests that it is important to consider not only the characteristics of individuals but also to examine the extent to which particular settings are able to facilitate the participation of community members. The purpose of this study was to understand the participation experiences of 54 Hmong refugees living in multiethnic housing developments. Interviews revealed that while Hmong residents valued participation highly, most were excluded from meaningful avenues of participation because of multiple barriers, including language differences, time constraints, and discrimination. No supports to address these barriers existed in their communities. It is important to understand and build individuals' capacities to participate and communities' capacities to promote involvement, integrate diversity, and foster interdependence. Participation is fundamental to the process of enabling refugees to become an integral part of their new communities and is a potential way for them to regain a sense of control over their lives and the decisions that affect them. © 2002 Wiley Periodicals, Inc.
Framing communication: Communicating the antismoking message effectively to all smokers
Authors: Wong Carissa O. , McMurray Nancy E.
Published: October 2002

There is a need to effectively communicate the antismoking issue across all sectors of the community, particularly to smokers with no intentions to quit smoking. Information can be framed using positively or negatively focused language to communicate the message content with maximum effect. Current cigarette smokers, with and without intentions to quit smoking (N = 70), were asked to respond to a Quit Smoking message framed either positively (benefits of quitting) or negatively (costs of not quitting). Participants were assessed pre‐post message and at 3‐months follow‐up. Results obtained suggest message frames had differential effects on message processing and increasing smokers' self‐confidence to quit smoking. In particular, trends in results suggest smokers with and without intentions to quit smoking interacted differently to framed messages. These findings may facilitate the improvement of community‐based health communications, by encouraging the use of differential message frames to effectively communicate the antismoking information to encourage all smokers to stop smoking and discourage others from adopting the smoking behavior. © 2002 Wiley Periodicals, Inc.
Anti‐proliferative effects of 8‐chloro‐cAMP and other cAMP analogs are unrelated to their effects on protein kinase A regulatory subunit expression
Authors: Lamb Darija , Steinberg Robert A.
Published: October 2002

Conflicting reports have attributed 8‐chloro‐cAMP (Cl‐cAMP)‐mediated inhibition of tumor cell growth to either a toxic 8‐chloro‐adenosine (Cl‐AdR) breakdown product or a Cl‐cAMP‐mediated decrease in ratio of Type I to Type II regulatory (R) subunits of protein kinase A (PKA). Using the MCF‐7 human breast cancer and S49 mouse lymphoma cell lines as models, we show that the effects of Cl‐cAMP and other cAMP analogs on growth and R subunit expression are unrelated. MCF‐7 cell growth was insensitive to most analogs and inducers of cAMP, but was potently inhibited by Cl‐cAMP acting through uptake and phosphorylation of its Cl‐AdR breakdown product. Possible roles of adenosine receptors or P2 purinoceptors in these Cl‐cAMP‐mediated growth effects were ruled out by studies with agonists and antagonists. Cholera toxin markedly decreased the ratio of Type I to Type II R subunits in MCF‐7 cells without affecting growth, while growth inhibitory concentrations of Cl‐cAMP or Cl‐AdR had insignificant effects on this ratio. In S49 cells, where PKA activation is known to inhibit cell growth, PKA‐deficient mutants retained sensitivity to both Cl‐cAMP and the related 8‐bromo‐cAMP. Adenosine kinase (AK)‐deficient S49 cells were inhibited only by higher concentrations of these 8‐halogenated cAMP analogs. Of the commonly used cAMP analogs, only 8‐(4‐chlorophenylthio)‐cAMP acted purely as a cyclic nucleotide—having no effect on PKA‐deficient cells, but strongly inhibiting both wild‐type and AK‐deficient cells. Where growth inhibitory concentrations of most cAMP analogs reduced RI expression in the AK‐deficient mutant, a functionally equivalent concentration of (N6, O2′)dibutyryl‐cAMP maintained or increased this expression. © 2002 Wiley‐Liss, Inc.
Oxidative stress in phagocytes—“The enemy within”
Authors: Splettstoesser Wolf D. , Schuff‐Werner Peter
Published: October 2002

Phagocytes represent a powerful defense system against invading microorganisms that threaten the life or functional integrity of the host. The capacity to generate and release substantial amounts of reactive oxygen species is a unique property of activated polymorphonuclear and mononuclear phagocytes. The crucial role of these molecules in killing microorganisms and their consecutive contribution to tissue damage during injury and inflammation is widely known. Although much research has been done to explore the molecular events involved in the interaction of oxygen intermediates with microbes or host tissue, surprisingly little attention has been paid to the effect of reactive metabolites on the phagocyte itself. This fact is especially surprising, since it is apparent that the activated phagocyte is directly exposed to its own toxic metabolites. The potential damage occurring during excessive radical formation might notably alter the vital functions of these primarily immunocompetent cells. Moreover, the critical role of oxygen radicals in apoptosis of leukocytes has been recently revealed. Apoptosis is now supposed to represent a key mechanism in neutrophil deactivation and resolution of inflammation. Therefore, this review will focus on the delicate balance between released oxidants and antioxidative protection within the phagocytes themselves. General and phagocyte‐specific antioxidative mechanisms, which have co‐evolved with the radical generating machinery of phagocytes, are discussed, since the outcome of local inflammation can directly depend on this antioxidative capacity and might range from adequate elimination of the pathogen with minimal acute tissue damage to progression towards a systemic inflammatory response syndrome. Microsc. Res. Tech. 57:441–455, 2002. © 2002 Wiley‐Liss, Inc.
How local irregularity gets global in a graph
Authors: Rautenbach Dieter , Volkmann Lutz
Published: October 2002

We prove asymptotically tight bounds on the difference between the maximum degree and the minimum degree of a simple graph in terms of its order and of the maximum difference between the degrees of adjacent vertices. Examples showing tightness and a conjecture are presented. © 2002 Wiley Periodicals, Inc. J Graph Theory 41: 18–23, 2002
Synthesis of tritium labeled isotopomers of L‐tyrosine
Authors: Augustyniak W. , Suchecki P. , Jemielity J. , Kański R. , Kańska M.
Published: October 2002

The synthesis of four selectively labeled isotopomers of L‐tyrosine, (L‐Tyr), using chemical and enzymatic methods is reported. Four tritium labeled isotopomers of L‐phenylalanine (L‐Phe) – [2‐3H]‐, [2′,6′‐3H2]‐, [3R‐3H]‐ and [3S‐3H]‐ have been synthesized using a combination of chemical and enzymatic methods. The labeled isotopomers of L‐Phe have been converted into [2 ‐3H]‐, [2′,6′‐3H2]‐, [3R‐3H]‐, and [3S‐3H]‐L‐Tyr by using the enzyme L‐phenyl‐alanine 4′‐monooxygenase. Copyright © 2002 John Wiley & Sons, Ltd.
Comparative studies of the development and differentiation of chloride cells in tilapine fish with different reproductive styles
Authors: Fishelson Lev , Bresler Vladimir
Published: October 2002

Using light and electron microscopy and fluorescent probes, we followed the ontogenesis of selected organs in embryos of several species of tilapia (Cichlidae, Pisces) with emphasis on chloride cell differentiation in species with two different reproductive styles: we compared the substrate‐brooder Tilapia zillii and the mouth‐brooders Oreochromis niloticus, O. aureus, Sarotherodon galilaeus, and Tristramella sacra. In all species a transitory blood network system nurtured by the vena caudalis inferiores supplied the yolk sac and preanal finfold during the advanced stages of embryonic and initial stages of larval development. During these stages chloride cells occurred on the yolk sac, as a part of the abdominal epithelium. The cells and their associated blood plexus remained active here until the gill‐lamellae, operculum, and mouth became functional. The chloride cells of their epithelium and blood system then took over, concomitant with a gradual degradation of the transitory blood system on the yolk sac. Ontogenesis of these systems (transitory and permanent) progressed at a higher rate in substrate‐brooders than in mouth‐brooders and was correlated with the earlier functioning of the gill‐operculum system. Thus, at a constant temperature of 26°C, the more exposed T. zillii progeny completed metamorphosis at 7–8 days after fertilization, calculated around 5,000 ± 80 h/temp, whereas juveniles of more protected mouth‐brooders attained a similar stage only 15 ± 1 days after fertilization and around 9,000 ± 200 h/temp. This earlier development of chloride cells and other pivotal organs in environmentally exposed progeny of substrate‐brooders, as compared to the protected progeny of mouth‐brooders, shows that their ontogeny was selected for the optimal survival style under specific etho‐ecological conditions. J. Morphol. 253:118–131, 2002. © 2002 Wiley‐Liss, Inc.
Ephemeroptera egg chorion characters: A test of their importance in assessing phylogenetic relationships
Authors: Domínguez Eduardo , Gabriela Cuezzo María
Published: October 2002

The egg chorion ultrastructures of the Hermanella–Traverella (Insecta: Ephemeroptera) species complex were studied from a comparative point of view and used for the first time in a cladistic analysis. Egg characters, along with other nymphal and adult morphological characters, were used to assess the phylogenetic relationships of the species complex. In order to test the value of egg characters, analyses were performed on three matrices: 1) egg characters alone, 2) adult and nymphal characters, and 3) adult, nymphal, and egg characters. The computer program Pee‐Wee was used to carry out the analysis. The cladistic analysis confirmed the value and potential of egg chorionic characters in assessing the phylogenetic relationships among ephemeropteran species. Egg characters, when added to the nymphal and adult character matrix, provided extra support to the monophyletic nature of the Hermanella–Traverella complex. Previously weakly defined clades were also resolved based on the new evidence. In the species studied the egg chorionic structures as well as their shape did not change after oviposition or water immersion, remaining constant through the different maturation stages of each species (mature nymph, subimago, and imago). For this reason, the eggs are a valuable source of information to unambiguously identify and associate a nymph to its correspondent adult stage when rearing is not possible. J. Morphol. 253:148–165, 2002. © 2002 Wiley‐Liss, Inc.
Serous cutaneous glands in new world hylid frogs: An ultrastructural study on skin poisons confirms phylogenetic relationships between Osteopilus septentrionalis and Phrynohyas venulosa
Authors: Delfino Giovanni , Brizzi Rossana , Nosi Daniele , Terreni Alessandro
Published: October 2002

Transmission electron microscope investigations of the serous (poison) skin glands in the New World tree frogs Osteopilus septentrionalis and Phrynohyas venulosa revealed that they produce granules with closely similar substructures, namely, a dense cortex and pale medulla. In both species these features, that contrast the complex, sometimes repeating patterns described in other hylid frogs, derive from similar secretory and maturational processes starting from the Golgi phase of poison biosynthesis. Observations on secretory discharge showed that the two species share common release mechanisms, based on bulk discharge (holocrine) processes. Our data provide novel evidence of the extensive ultrastructural polymorphism of serous skin products in Hylidae and agree with phylogenies that regard this family as polyphyletic in origin. Assuming that ultrastructural features of cutaneous poison biosynthesis and maturation are adequate clues for tracking anuran phylogeny, the present findings also support a close relationship between Osteopilus and Phrynohyas taxa as previously suggested by osteological evidence. J. Morphol. 253:176–186, 2002. © 2002 Wiley‐Liss, Inc.
In vitro susceptibility to infection with SIVcpz and HIV‐1 is lower in chimpanzee than in human peripheral blood mononuclear cells*
Authors: Ondoa Pascale , Davis David , Kestens Luc , Vereecken Chris , Ribas Sergio Garcìa , Fransen Katrien , Heeney Jonathan , van der Groen Guido
Published: October 2002

This study was undertaken to evaluate and compare the susceptibility of chimpanzee versus human peripheral blood mononuclear cells (PBMCs) to infection with SIVcpz and HIV‐1 non‐syncitium inducing primary isolates. The results demonstrate clearly that chimpanzee PBMCs have a lower capacity to support viral replication as compared to human PBMCs. There was no experimental evidence that this difference was due to a lower availability of target cells for viral infection (PBMCs positive for CD4 and CCR5 molecules) or to a differential susceptibility to apoptosis (PBMCs positive for CD4 and CD95 molecules). A lower capacity of chimpanzee PBMCs to support SIVcpz and HIV‐1 replication in vitro is related to a post‐entry barrier to virus replication. J. Med Virol. 67:301–311, 2002. © 2002 Wiley‐Liss, Inc.
Detection of parvovirus B19 NS1‐specific antibodies by ELISA and western blotting employing recombinant NS1 protein as antigen
Authors: Heegaard Erik D. , Rasksen Cecilie J. , Christensen Jesper
Published: October 2002

Human parvovirus B19 (B19) encodes a number of nonstructural proteins, including the major protein, NS1, and two structural proteins, VP1 and VP2. The use of denatured NS1 in enzyme‐linked immunosorbent assay (ELISA) and Western blot (WB) assay has provided an opportunity to study some of the immunologic properties of NS1, but the results have been equivocal and the diagnostic sensitivity poor, probably because of the absence of conformational epitopes. Various viral isolates and baculovirus vectors were employed to produce recombinant B19 NS1 under nondenaturing conditions for the first time. To assess the antigenicity of purified B19 NS1, the reaction patterns of 252 samples were compared by B19 NS1 and VP2 ELISA. In sera from individuals with past infection (VP2 IgG‐positive), the use of this new antigen increased significantly the sensitivity of ELISA compared with WB (78% vs. 33%, P = 0.001), contradicting perpetuated claims that B19 NS1 IgG is detected primarily in patients with arthralgia or chronic infection. Previous reports of the absence of NS1 IgG during the initial phase of infection (< 6 weeks) were proved incorrect by the detection of NS1 IgG in 60% of samples from patients recently infected by B19. Including conformational epitopes in the ELISA increases the diagnostic sensitivity, although immunologically, a temporal (years) attenuation of NS1 antibodies appears to take place. This novel diagnostic tool may be useful as a supplement in case of borderline results by VP2 ELISA and for monitoring the efficacy of future capsid‐based B19 vaccines. J. Med Virol. 67:375–383, 2002. © 2002 Wiley‐Liss, Inc.
Strong interaction between human herpesvirus 6 and peripheral blood monocytes/macrophages during acute infection
Authors: Kondo Kazuhiro , Kondo Toshio , Shimada Kazuya , Amo Kiyoko , Miyagawa Hiromi , Yamanishi Koichi
Published: October 2002

Human herpesvirus 6 (HHV‐6) encodes a viral chemokine and chemokine receptors that may modify the functions of monocytes/macrophages (MO/Mϕ) during productive HHV‐6 infection. The interactions between HHV‐6 and MO/Mϕ during acute infection, however, remain poorly understood. In this study, we investigated the tropism of HHV‐6 in peripheral blood mononuclear cells (PBMCs) during acute infection. We detected 637 ± 273 copies of viral DNA in 104 MO/Mϕ. in contrast, in 104 CD4+ T cells, which have been reported to be viral carriers during the acute infection of HHV‐6, we found only 115 ± 42 copies of viral DNA. Consistent with these data, virus was isolated from MO/Mϕ an order of magnitude more frequently than from CD4+ T cells. Viral mRNA U79/80, which indicates viral replication, was detectable in the MO/Mϕ. In addition, the mRNAs that encode viral chemokine receptors U12 and U51, which may modify the function of MO/Mϕ, were expressed in the cells. Therefore, productively infected MO/Mϕ may be the dominant cell population that is responsible for HHV‐6 viremia during acute HHV‐6 infection. The strong interaction of HHV‐6 with MO/Mϕ may be partly responsible for the pathogenesis of this virus. J. Med Virol. 67:364–369, 2002. © 2002 Wiley‐Liss, Inc.
Prevention and control of hepatitis B in China
Authors: Sun Zongtang , Ming Lihua , Zhu Xu , Lu Jianhua
Published: October 2002

About 170 million Chinese are infected chronically with HBV and 10% suffer from chronic hepatitis. Around half a million Chinese die from hepatitis B caused hepatocellular carcinoma and endstage cirrhosis each year. From 1983 to the present, a controlled clinical trial involving 80,000 children on a universal hepatitis B vaccination programme to prevent chronic hepatitis, hepatocellular carcinoma, and endstage cirrhosis was implemented in Qidong, China. A pilot study demonstrated that the HBsAg rate reached the adult level before the fifth year of age, and neonatal vaccination with either plasma‐derived or recombinant hepatitis B vaccines provided a similar 75% protective efficacy against HBV infection. The high rate of follow‐up and blood tests coverage of the cohorts provided data to show 75% protection at the tenth to eleventh years of age against serum HBsAg and also against prolonged hepatic dysfunction. The strategy of controlling hepatitis B nationwide was based on the universal immunisation of newborns, beginning in cities and then the rural areas. The large‐scale vaccine source was provided by domestic plants through technology transfer, first providing plasma‐derived vaccine replaced completely by recombinant DNA vaccine in 1997. An official survey in 1999 using a cluster sampling of 25,878 children from 31 provinces reported an average coverage rate of three dose of hepatitis B vaccination of 70.7%, being higher in urban areas. The Ministry of Public Health of China has planned to integrate hepatitis B vaccination into the nationwide EPI program with Government‐provided vaccines starting January 1, 2002. J. Med. Virol. 67:447–450, 2002. © 2002 Wiley‐Liss, Inc.
Immunoglobulin G antibody avidity assay for serodiagnosis of hepatitis C virus infection
Authors: Kanno Atsushi , Kazuyama Yukumasa
Published: October 2002

It has been reported that the avidity of specific IgG antibody is lower in primary viral infection than in chronic viral infection. However, few studies have been reported on the IgG avidity in hepatitis C virus (HCV) infection. In the present study, 36 patients with antibody to HCV (anti‐HCV) were examined for IgG avidity by an enzyme immunoassay with or without urea elution. The avidity index was significantly low in patients with primary HCV infection (7.7 ± 6.8%, mean ± SD), compared with patients with chronic HCV infection (77.0 ± 21.8%) and individuals with past HCV infection (44.5 ± 12.6%). Temporal changes of IgG avidity were examined in six patients with primary HCV infection. The avidity index was low in the acute phase of the infection and then increased with time. These results suggest that the avidity assay for IgG anti‐HCV is a useful method for distinguishing primary HCV infection from chronic or past HCV infection. J. Med. Virol. 68:229–233, 2002. © 2002 Wiley‐Liss, Inc.
Variation of hepatitis C virus load, hypervariable region 1 quasispecies and CD81 hepatocyte expression in hepatocellular carcinoma and adjacent non‐cancerous liver
Authors: Young Kung‐Chia , Lin Pin‐Wen , Hsiao Wei‐Chiang , Chang Ting‐Tsung , Chang Yu‐Chung , Wu Hwa‐Lin
Published: October 2002

Hepatitis C virus (HCV) infection is etiologically associated with the development of hepatocellular carcinoma (HCC) worldwide. HCV has been reported to exist and replicate in both HCC and adjacent non‐cancerous liver tissue, but limited information was available on HCV viral load and quasispecies composition in HCC relative to adjacent non‐cancerous hepatocytes. Previous study has also suggested CD81, a surface hepatocyte protein, as a receptor for HCV. To clarify the above, HCV‐RNA and CD81‐RNA titers in 20 paired hepatectomized liver and serum were quantitatively measured by chemiluminescent RT‐cPCR. Hypervariable region 1 (HVR‐1) variations of parallel specimens were analyzed after subcloning in 6 patients. HCV‐RNA levels in serum and non‐cancerous liver were markedly higher for HCV genotype 1 than genotype non‐1. HCV levels were markedly higher in non‐cancerous liver than in HCC (P = 0.001) in a genotype‐independent manner, with a mean ratio of 56:1 for non‐cancerous tissue to HCC. Both non‐cancerous and HCC tissues had the same level of CD81‐RNA expression, which was not linked to HCV load. HCV‐RNA quantity in both HCC and non‐cancerous liver correlated with the number of HVR‐1 quasispecies in the tissue, and distinct HVR‐1 subclones existed. J. Med. Virol. 68:188–196, 2002. © 2002 Wiley‐Liss, Inc.
Replication of enhancer‐deficient amphotropic murine leukemia virus in human fibrosarcoma but not in primary human fibroblasts
Authors: Reuss Frank U. , Berdel Bianca , Heber Ricarda , Bantel‐Schaal Ursula
Published: October 2002

Amphotropic murine leukemia virus (MLV) replicates in cells from various mammalian species including humans and is a potential contaminant in MLV vector preparations for human gene transfer studies. In general, MLV replication depends on the expression of viral genes under the control of 75 bp enhancer elements in the long terminal repeat. However, in specific human fibrosarcoma and lymphoma lines replication of amphotropic MLV is possible without these enhancers. Fibrosarcomas are malignant tumors of fibroblast origin. To test the replication potential of intact and enhancerless amphotropic MLV in untransformed cells, infection studies with these viruses were carried out in three types of primary human fibroblasts. Replication of amphotropic MLV is observed in two of three tested fibroblast strains. None of these primary human fibroblasts is permissive for enhancer‐deficient MLV, suggesting that replication of this virus may be limited to transformed cells. J. Med. Virol. 68: 278–284, 2002. © 2002 Wiley‐Liss, Inc.
Transition probability coefficients and the stability of finite difference schemes for the diffusion and Telegraphers' equations
Authors: Malachowski Michal J.
Published: October 2002

A probabilistic approach has been used to analyse the stability of the various finite difference formulations for propagation of signals on a lossy transmission line. If the sign of certain transition probabilities is negative, then the algorithm is found to be unstable. We extend the concept to consider the effects of space and time discretizations on the signs of the coefficients in a probabilistic finite difference implementation of the Telegraphers' equation and draw parallels with the transmission line matrix (TLM) technique. Copyright © 2002 John Wiley & Sons, Ltd.
TLM for diffusion: consistent first time step. Two‐dimensional case
Authors: Enders Peter , H. Pulko Susan , Stubbs David M.
Published: October 2002

In initializing a transmission line matrix (TLM) diffusion model it is necessary to consider both initial concentration (temperature) and initial flow. As usual, only one of them is given; an auxiliary formula is necessary to calculate the distribution for the first time step. It has been shown that the standard formula may introduce additional numerical errors (International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 1993; 6:135; International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 1993; 6:161) and that these errors can persist over many time steps. In this paper, we show how an initial modification to the normal TLM algorithm can remove such errors, and we demonstrate the method by applying it to a two‐dimensional TLM heat diffusion model for a copper plate. Copyright © 2002 John Wiley & Sons, Ltd.
TLM model of waves in moving media
Authors: O'Connor William J.
Published: October 2002

Waves in moving media arise in fluid mechanics and elastic mechanics and are especially significant in acoustics. They are characterized by direction‐dependent wave speeds. When compared with the wave equation, the governing differential equation, ytt+αyxt+βyxx=0 has an extra, cross derivative term. An original solution method based on transmission line matrix (TLM) modelling is presented. By means of supplementary link lines controlled by notional ‘diodes’ the TLM mesh wave propagation characteristics are directionally biased. The algorithm is derived and performance assessed for the one‐dimensional case. Numerical results agree closely with analytical solutions. The algorithm has interesting properties promising new application areas for TLM. Copyright © 2002 John Wiley & Sons, Ltd.