bluuebunny/crossref_metadata_2025 huggingface dataset:
Lack of ifn‐γ‐mediated induction of the class II transactivator (CIITA) through promoter methylation is predominantly found in developmental tumor cell lines
Authors: van der Stoep Nienke , Biesta Paula , Quinten Edwin , van den Elsen Peter J.
Published: August 2002

Downregulation of major histocompatibility complex (MHC) molecules by tumor cells impairs cellular immune recognition and contributes to inefficient cell‐mediated tumor eradication. Low or lack of expression of MHC molecules is frequently observed in early developmental or embryonically derived tumor cells. Considering the central role of the class II transactivator (CIITA) in MHC class II‐ and class I‐mediated antigen presentation, we compared the induction of CIITA by interferon‐γ (IFN‐γ) in a diverse panel of developmental and more differentiated tumor cell lines. In contrast to the more differentiated tumor cell lines, none of the developmental tumor cell lines were capable of expressing CIITA after treatment with IFN‐γ. Remarkably, in transient transfection assays, CIITA promoter IV (CIITA‐PIV) was found to be induced by IFN‐γ. Southern blot analysis of genomic DNA obtained from the developmental tumor cell lines indicated that the absence of endogenous CIITA induction was due to methylation of the CIITA‐PIV region. Exposure to 5‐azacytidine restored induction of CIITA and congruent HLA‐DRA expression in these cells. The observation that only developmental tumor cell lines, originating from various tissues, employ methylation to silence CIITA expression may reflect the natural status of CIITA expression during early development rather than oncogenic transformation. © 2001 Wiley‐Liss, Inc.
Demographic responses to environmental stress in the drought‐ and famine‐prone areas of northern Ethiopia
Authors: Ezra Markos
Published: August 2002

Research on the demographic consequences of environmental stress (drought and famine, in particular) has not been conclusive. Using primary survey data on the demographic consequences of drought and famine in the drought‐prone areas of northern Ethiopia, this study argues that growing environmental stress and persisting food insecurity have stimulated changes in the demographic behaviours and attitudes of farming communities, including: an increase in acceptance rates of family planning services; changing attitudes towards early marriage and having a large number of children; actual reduction in fertility; migration (particularly of the youth) out of the communities; and the tendency by many farmers to be involved in non‐farm income‐generating activities and employment sectors. Copyright © 2001 John Wiley & Sons, Ltd.
Regional fertility differences in western Germany: an overview of the literature and recent descriptive findings
Authors: Hank Karsten
Published: August 2002

In the years following German unification, researchers' attention has been attracted mainly by the rapid fertility decline in eastern Germany. This has often resulted in neglect of regional fertility differentials within Germany, which reach much further back in time than those currently observed between the ‘old’ and the ‘new’ Länder of the Federal Republic. This paper provides an overview of studies investigating regional fertility differences in post‐war western Germany. Additionally, results of an analysis of official statistics for western German Kreise (districts) in the period 1995–97 are presented. Geographical patterns of high‐ and low‐fertility areas are detected, which have remained basically unchanged by the general fertility decline of the late 1960s and early 1970s. Consistent with findings from earlier studies, population density, family migration and the local occupational structure are still found to be closely associated with regional total fertility rates in the 1990s. The paper concludes with a call for more multilevel studies that –theoretically as well as empirically –link regional social contexts with individual (reproductive) behaviour. Copyright © 2001 John Wiley & Sons, Ltd.
Alcohol and drug‐related effects on development: A new emphasis on contextual factors
Authors: Jacobson Sandra W. , Jacobson Joseph L.
Published: August 2002

The articles in this special issue bring together two lines of research that reflect different perspectives on the effects of parental alcohol and drug abuse—the long‐term impact of teratogenic insult due to prenatal exposure and the developmental risks associated with being reared by a substance‐abusing parent. These articles reflect major methodological advances, including the use of moderator variables—particularly, parental psychopathology and child temperament—for detecting the adverse impact of parental substance abuse on child outcome. In several articles, more comprehensive sets of control variables are considered than in prior studies, and investigators examine the use of aggregate and cumulative risk factors in an attempt to better model underlying conditions of risk. Another advance is that most of these studies recognize the importance of quantifying levels of prenatal alcohol and drug exposure. In focusing our attention simultaneously on prenatal exposure, parental substance abuse and comorbid psychopathology, child temperament, and contextual factors, these studies offer a better understanding of the complex interrelated influences of risk factors on the child's cognitive and socio‐emotional development. © 2001 Michigan Association for Infant Mental Health.
Serially correlated variables in dynamic, discrete choice models
Authors: Stinebrickner Todd R.
Published: August 2002

This paper discusses the problems that are encountered when dynamic, discrete choice models are specified with continuous, serially correlated state variables. A variety of approximation methods that can deal with these problems is examined, and an empirical example that allows continuous variables to be serially correlated is presented. Copyright © 2000 John Wiley & Sons, Ltd.
A multivariate latent factor decomposition of international bond yield spreads
Authors: Dungey Mardi , Martin Vance L , Pagan Adrian R
Published: August 2002

A factor analysis of long‐term bond spreads is performed by decomposing international interest rate spreads into national and global factors. The factors are latent, and are assumed to have GARCH‐type specifications as well as exhibiting serial dependence. An indirect estimator is used to compute estimates of the unknown parameters. The sampling performance of this estimator is investigated and compared with an alternative direct estimator based on the Kalman predictor. The factor model is applied to weekly data on long‐bond spreads between five countries ‐ Australia, Japan, Germany, Canada and the UK ‐ and the USA over the period 1991 to 1999. The resulting factor decomposition is used to examine the international investor's optimal portfolio decision in a mean‐variance framework. Copyright © 2000 John Wiley & Sons, Ltd.
Bayesian semiparametric estimation of discrete duration models: an application of the dirichlet process prior
Authors: Campolieti Michele
Published: August 2002

This paper proposes a Bayesian estimator for a discrete time duration model which incorporates a non‐parametric specification of the unobserved heterogeneity distribution, through the use of a Dirichlet process prior. This estimator offers distinct advantages over the Nonparametric Maximum Likelihood estimator of this model. First, it allows for exact finite sample inference. Second, it is easily estimated and mixed with flexible specifications of the baseline hazard. An application of the model to employment duration data from the Canadian province of New Brunswick is provided. Copyright © 2001 John Wiley & Sons, Ltd.
Trend‐stationary GNP: evidence from a new exact pointwise most powerful invariant unit root test
Authors: Shively Philip A.
Published: August 2002

There has been a substantial debate whether GNP has a unit root. However, statistical tests have had little success in distinguishing between unit‐root and trend‐reverting specifications because of poor statistical properties. This paper develops a new exact small‐sample, pointwise most powerful unit root test that is invariant to the unknown mean and scale of the time series tested, that generates exact small‐sample critical values, powers and p‐values, that has power which approximates the maximum possible power, and that is highly robust to conditional heteroscedasticity. This test decisively rejects the unit root null hypothesis when applied to annual US real GNP and US real per capita GNP series. This paper also develops a modified version of the test to address whether a time series contains a permanent, unit root process in addition to a temporary, stationary process. It shows that if these GNP series contain a unit root process in addition to the stationary process, then it is most likely very small. Copyright © 2001 John Wiley & Sons, Ltd.
Combined pulmonary toxicity of cadmium chloride and sodium diethyldithiocarbamate
Authors: Tátrai Erzsébet , Brózik Márta , Náray Miklós , Adamis Zoltán , Ungváry György
Published: August 2002

The pulmonary toxicity of sodium diethyldithiocarbamate and cadmium chloride, each separately and in combination, was compared in Sprague‐Dawley rats after single intratracheal instillation in sequential experiments by chemical, immunological and morphological methods. With combined exposure, the cadmium content of the lungs increased permanently relative to that of the lungs of just cadmium‐treated animals. Immunoglobulin levels of the whole blood did not change, whereas in bronchoalveolar lavage the IgA and IgG levels increased significantly. Morphological changes were characteristic of the effects of cadmium but were more extensive and more serious than in the case of cadmium administration alone: by the end of the first month, interstitial fibrosis, emphysema and injury of membranes of type I pneumocytes developed and hypertrophy and loss of microvilli in type II pneumocytes were detectable. These results showed that although dithiocarbamates as chelating agents are suitable for the removal of cadmium from organisms, they alter the redistribution of cadmium within the organism, thereby increasing the cadmium content in the lungs, and structural changes are more serious than observed upon cadmium exposure alone. Copyright © 2001 John Wiley & Sons, Ltd.
In vitro skin absorption and decontamination of sulphur mustard: comparison of human and pig‐ear skin
Authors: Chilcott R. P. , Jenner J. , Hotchkiss S. A. M. , Rice P.
Published: August 2002

The aim of this study was to evaluate the use of an in vitro skin diffusion cell system as a model for assessing decontaminants against the chemical warfare agent sulphur mustard (SM). The in vitro absorption rates of SM through heat‐separated human (157 ± 66 µg cm−2 h−1) and pig‐ear (411 ± 175 µg cm−2 h−1) epidermal membranes were in agreement with previous in vivo studies that quoted skin absorption rates of 150 and 366 mu g cm−2 h−1, respectively.

Decontaminants (fuller's earth, Ambergard and BDH spillage granules) were ranked in order of effectiveness by measuring the skin absorption rates and the percentage of applied dose of SM that penetrated human and pig‐ear epidermal membranes. The effectiveness of fuller's earth measured in this in vitro study using human epidermal membranes was in agreement with a previous in vivo human volunteer study. Similarly, the effectiveness of fuller's earth and Ambergard measured in vitro with pig‐ear epidermal membranes was in agreement with a previous in vivo study conducted on rats. However, there was complete disparity in the ranking of decontaminants between human and pig‐ear epidermal membranes measured in vitro.

Thus, although pig‐ear skin may be a relatively good model for predicting the human skin absorption of SM, it is a poor model for testing decontamination systems. The results of this study further validate the use of Franz‐type glass diffusion cells containing human epidermal membranes as a model for predicting in vivo human skin absorption. Copyright © 2001 John Wiley & Sons, Ltd.
Oxidative preconditioning affords protection against carbon tetrachloride‐induced glycogen depletion and oxidative stress in rats
Authors: Candelario‐Jalil E. , Mohammed‐Al‐Dalain S. , Fernández O. S. León , Menéndez S. , Pérez‐Davison G. , Merino N. , Sam S. , Ajamieh H. H.
Published: August 2002

The rectal insufflation of a judicious dose of ozone, selected from that used in clinical practice, is able to promote oxidative preconditioning or oxidative stress tolerance preventing the hepatocellular damage mediated by free radicals. In order to evaluate the effects of ozone oxidative preconditioning on carbon tetrachloride‐mediated hepatotoxicity, the following experimental protocol was designed: group 1 (negative control, sunflower oil i.p.); group 2 (CCl4 in sunflower oil, 1 ml kg−1 i.p.); group 3 (15 ozone–oxygen pretreatments at a dose of 1 mg kg−1 via rectal insufflation + CCl4 as in group 2); group 4 (ozone control group, 15 ozone–oxygen pretreatments + sunflower oil i.p.). Ozone pretreatment prevented glycogen depletion (as demonstrated by biochemical and histopathological findings) and avoided lactate overproduction associated with the hepatotoxic effects of CCl4. The administration of CCl4 increased lipid peroxidation (as measured by thiobarbituric acid‐reactive substances) and uric acid levels and inhibited superoxide dismutase activity. All these deleterious effects induced by CCl4 were prevented by ozone pretreatment. The administration of ozone without CCl4 (ozone control group) did not produce any changes in the evaluated parameters. Our results showed that ozone treatment, in our experimental conditions, was able to prevent anaerobic glycolysis and oxidative stress induced by CCl4. Copyright © 2001 John Wiley & Sons, Ltd.
Sulfur mustard‐stimulated protease: a target for antivesicant drugs
Authors: Ray P. , Chakrabarti A. K. , Broomfield C. A. , Ray R.
Published: August 2002

One of the mechanisms of the skin blistering effect (vesication) of sulfur mustard (bis‐(2‐chloroethyl)sulfide, HD) is believed to be via the stimulation of specific protease(s) at the dermal–epidermal junction. Cultured normal human epidermal keratinocytes (NHEK) were used as a model to study and characterize protease stimulated by the mustards 2‐chloroethyl ethyl sulfide (CEES), 2‐chloro‐N‐(2‐chloroethyl)‐N‐methylethanamine hydrochloride (nitrogen mustard, HN2) and HD. The results obtained using a chromozym (TRY) peptide substrate protease assay revealed the optimum mustard concentrations and time for protease stimulation to be about 200 µM (CEES), 100 µM(HN2) and 100 µM (HD) and 16 h. The mustard‐stimulated protease was membrane bound and was inhibited by adding a Ca2+ chelator (either 2 mM EGTA (ethylene glycol‐bis(amino ethyl ether) N,N,N′,N′ tetraacetic acid) or 50 µM BAPTA AM (1,2‐bis(z‐aminophenoxy)ethane‐N,N,N′,N′‐tetraacetic acid, tetraacetoxy methyl ester) alone or in combination), a serine protease inhibitor diisopropyl fluoro‐phosphate (DFP, 1 mM), or a protein synthesis inhibitor cycloheximide (35 µM) in the extracellular medium. These results suggest that mustard toxicity may involve the stimulation of trypsin/chymotrypsin‐like serine protease, dependent on Ca2+ and new protein synthesis. Protein purification by gel exclusion and hydrophobic chromatography produced a 70–80 kDa protease, which had an amino acid sequence homologous with a mammalian‐type bacterial serine endopeptidase. Based on this information, research is in progress to identify the protease stimulated by HD in NHEK and to determine whether its inhibitors are useful as prospective antivesicant drugs. Copyright © 2002 John Wiley & Sons, Ltd.
Dose‐dependent behavior of bioactive glass dissolution
Authors: Jones Julian R. , Sepulveda Pilar , Hench Larry L.
Published: August 2002

The effect of glass dosage (0.001 g ml−1 to 0.015 g ml−1) on the in vitro dynamic dissolution behavior of melt‐derived 45S5 and sol‐gel–derived 58S bioactive glasses, in simulated body fluid (SBF) at 37 °C, was evaluated. These glasses differ significantly in texture, especially the specific surface area and porosity, as a result of differences in manufacturing route. The concentrations of elements (Si, Ca, P, and Na) leached from the glasses into the dissolution medium, from 1 to 22 h, were evaluated with the use of induced coupled plasma analysis (ICP). The reacted powders were analyzed with the use of FTIR to observe the formation of a hydroxycarbonate apatite layer on the surface. The results show that the rate of HCA formation on both gel‐ and melt‐derived bioactive glass powders in vitro depends on the concentration of the powders in solution. This result must be taken into account when carrying out in vitro cell‐culture studies to simulate conditions in vivo and in experiments using extracts of the bioactive glass powders. © 2001 John Wiley & Sons, Inc. J Biomed Mater Res (Appl Biomater) 58: 720–726, 2001
Effect of fibrin sealant composition on human neutrophil chemotaxis
Authors: Hanson Angela J. , Quinn Mark T.
Published: August 2002

The use of fibrin sealants offers one of the most physiologically compatible approaches to preventing postoperative adhesions. Although a number of fibrin sealant formulations have been developed, little is known about how the various components of these preparations affect the wound‐healing process. Because one of the key steps in wound healing is the migration of phagocytic leukocytes, such as neutrophils, into the site of injury, we performed studies to characterize systematically the effects of various fibrin sealant components on neutrophil chemotaxis. Using a transwell chemotaxis assay, we found that increasing fibrin concentration resulted in an inhibition of the ability of the cells to migrate through the clots in a dose‐dependent manner, and at fibrin clot concentrations >2.0 mg/mL chemotaxis was completely blocked. Factor XIII crosslinking of the clots also had a significant impact on neutrophil chemotaxis, and sealant preparations deficient in Factor XIII allowed neutrophil migration at much higher fibrin concentrations. The presence of various other fibrin sealant components such as plasminogen and fibrinolysis inhibitors (aprotinin and tranexamic acid) did not have any significant effects on the ability of neutrophils to migrate through fibrin clots as compared to control clots without these components. Overall, these studies show that the composition of fibrin sealant preparations can significantly affect neutrophil migration into the site of injury, which could possibly influence the wound healing process. © 2002 Wiley Periodicals, Inc. J Biomed Mater Res 61: 474–481, 2002
Micromorphological changes in resin‐dentin bonds after 1 year of water storage
Authors: Hashimoto Masanori , Ohno Hiroki , Sano Hidehiko , Tay Franklin R. , Kaga Masayuki , Kudou Yoshiyuki , Oguchi Haruhisa , Araki Yoshima , Kubota Minoru
Published: August 2002

The purpose of this study was to evaluate the degradation of resin–dentin bonds after 1 year of water storage. Resin–dentin‐bonded specimens were prepared with the use of an adhesive resin system (One‐Step: Bisco). Half of the experimental specimens were sectioned perpendicular to the adhesive interface to produce a beam (adhesive area: 0.9 mm2) before being stored in distilled water at 37 °C for 1 year. The remaining half of the bonded specimens were sectioned into beams of similar dimensions after 1 year of water storage. Additional bonded specimens that had been stored in water for 24 h before sectioning into beams were used as controls. The beams in the two experimental groups and the control group were subjected to microtensile bond testing. Fractography was performed on all fractured beams with the use of FE‐SEM. There were significant (p < .05) differences in bond strength among the control specimens (55.9 ± 12.9 MPa), specimens that had been sectioned into beams after water storage (68.9 ± 18.6 MPa), and specimens that had been sectioned into beams before water storage (28.1 ± 9.3 MPa). Fractography revealed that the resin material was gradually extracted from the periphery to the center portion of the beam. This probably accounted for the decrease in bond strength after 1 year of water storage. © 2002 Wiley Periodicals, Inc. J Biomed Mater Res (Appl Biomater) 63: 306–311, 2002
Angiogenesis and neovascularization associated with extracellular matrix‐modified porous implants
Authors: Kidd Kameha R. , Nagle Raymond B. , Williams Stuart K.
Published: August 2002

Therapies directed toward stimulation of angiogenesis seek to accelerate the development of new blood vessels in tissues rendered dysfunctional because of an insufficient microvascular supply. The goal of the current study was the stimulation of an angiogenic response around and within porous biomedical implants, such as vascular grafts, constructed from a base polymer composed of expanded polytetrafluoroethylene (ePTFE). Similar to many biomaterials, ePTFE does not elicit a significant angiogenic response and the porous interstices of this material remain avascular after implantation. Studies were performed to evaluate the ability of a tumorigenic cell line, the 804‐G rat kidney cell to secrete an angiogenic extracellular matrix on and within the porous structures of ePTFE. A rat model was used to evaluate and compare implant‐associated healing responses between nonmodified materials and extracellular matrix‐modified ePTFE. Results demonstrated that, in contrast to untreated ePTFE, the matrix‐modified ePTFE stimulated both angiogenesis in implant‐associated tissue and neovascularization of the pores within the ePTFE interstices. Deposition of an insoluble matrix stimulates an angiogenic response and has a potential application for the improvement of medical device function. © 2001 Wiley Periodicals, Inc. J Biomed Mater Res 59: 366–377, 2002
Supercharging Your ABC: Use Revenue Data!
Authors: Brewer Peter C. , Campbell Robert J. , McClure Richard H.
Published: August 2002

As in most managerial decision making, activity‐based costing (ABC) requires that managers
first determine what information is relevant. Historically, the focus of ABC systems has been almost entirely on
costs, yet revenue‐oriented information is often highly relevant and useful. Incorporating both the revenue
and expense sides of the income statement can make ABC far more powerful. © 2002 Wiley Periodicals,
Inc.
CITGO Petroleum Meeting the Challenge with ABM
Authors: Rodriguez Marlene B. , Muras Andrew D. , Calhoun Dennis D.
Published: August 2002

Activity‐based management (ABM) has helped process and energy industries cut costs and improve
efficiencies, because it provides the baseline data needed for performance measurement, outsourcing, product
costing, and capacity analysis. However, implementations of ABM have to be tailored to meet the unique challenges
posed by Shared Services functions and operational units. This article outlines CITGO's implementation
techniques for ABM and describes the company's results for its Shared Services and operations business units.
© 2002 Wiley Periodicals, Inc.
How to Use Transactional Databases for M&A
Authors: Pratt Shannon P.
Published: August 2002

Want to price M&A transactions more accurately and structure them more favorably? Then take a look
at other completed transactions, advises the author. Databases now contain details on thousands of M&A
transactions. The author shows you how to use them. © 2002 Wiley Periodicals, Inc.
Remodeling of sperm chromatin after fertilization involves nucleosomes formed by sperm histones H2A and H2B and two CS histone variants
Authors: Oliver M. Isabel , Concha Carolina , Gutiérrez Soraya , Bustos Alejandra , Montecino Martín , Puchi Marcia , Imschenetzky María
Published: August 2002

The composition of nucleosomes at an intermediate stage of male pronucleus formation was determined in sea urchins. Nucleosomes were isolated from zygotes harvested 10 min post‐insemination, whole nucleoprotein particles were obtained from nucleus by nuclease digestion, and nucleosomes were subsequently purified by a sucrose gradient fractionation. The nucleosomes derived from male pronucleus were separated from those derived from female pronucleus by immunoadsorption to antibodies against sperm specific histones (anti‐SpH) covalently bound to Sepharose 4B (anti‐SpH‐Sepharose). The immunoadsorbed nucleosomes were eluted, and the histones were analyzed by Western blots. Sperm histones (SpH) or alternatively, the histones from unfertilized eggs (CS histone variants), were identified with antibodies directed against each set of histones. It was found that these nucleosomes are organized by a core formed by sperm histones H2A and H2B combined with two major CS histone variants. Such a hybrid histone core interacts with DNA fragments of approximately 100 bp. It was also found that these atypical nucleosome cores are subsequently organized in a chromatin fiber that exhibits periodic nuclease hypersensitive sites determined by DNA fragments of 500 bp of DNA. It was found that these nucleoprotein particles were organized primarily by the hybrid nucleosomes described above. We postulate that this unique chromatin organization defines an intermediate stage of male chromatin remodeling after fertilization. J. Cell. Biochem. 85: 851–859, 2002. © 2002 Wiley‐Liss, Inc.
Inhibition of nitric oxide synthase inhibitors and lipopolysaccharide induced inducible NOS and cyclooxygenase‐2 gene expressions by rutin, quercetin, and quercetin pentaacetate in RAW 264.7 macrophages
Authors: Chen Yen‐Chou , Shen Shing‐Chuan , Lee Woan‐Ruoh , Hou Wen‐Chi , Yang Ling‐Ling , Lee Tony J.F.
Published: August 2002

Several natural flavonoids have been demonstrated to perform some beneficial biological activities, however, higher‐effective concentrations and poor‐absorptive efficacy in body of flavonoids blocked their practical applications. In the present study, we provided evidences to demonstrate that flavonoids rutin, quercetin, and its acetylated product quercetin pentaacetate were able to be used with nitric oxide synthase (NOS) inhibitors (N‐nitro‐L‐arginine (NLA) or N‐nitro‐L‐arginine methyl ester (L‐NAME)) in treatment of lipopolysaccharide (LPS) induced nitric oxide (NO) and prostaglandin E2 (PGE2) productions, inducible nitric oxide synthase (iNOS) and cyclooxygenase‐2 (COX‐2) gene expressions in a mouse macrophage cell line (RAW 264.7). The results showed that rutin, quercetin, and quercetin pentaacetate‐inhibited LPS‐induced NO production in a concentration‐dependent manner without obvious cytotoxic effect on cells by MTT assay using 3‐[4,5‐dimethylthiazol‐2‐yl]‐2,5‐diphenyltetrazolium bromide as an indicator. Decrease of NO production by flavonoids was consistent with the inhibition on LPS‐induced iNOS gene expression by western blotting. However, these compounds were unable to block iNOS enzyme activity by direct and indirect measurement on iNOS enzyme activity. Quercetin pentaacetate showed the obvious inhibition on LPS‐induced PGE2 production and COX‐2 gene expression and the inhibition was not result of suppression on COX‐2 enzyme activity. Previous study demonstrated that decrease of NO production by L‐arginine analogs effectively stimulated LPS‐induced iNOS gene expression, and proposed that stimulatory effects on iNOS protein by NOS inhibitors might be harmful in treating sepsis. In this study, NLA or L‐NAME treatment stimulated significantly on LPS‐induced iNOS (but not COX‐2) protein in RAW 264.7 cells which was inhibited by these three compounds. Quercetin pentaacetate, but not quercetin and rutin, showed the strong inhibitory activity on PGE2 production and COX‐2 protein expression in NLA/LPS or L‐NAME/LPS co‐treated RAW 264.7 cells. These results indicated that combinatorial treatment of L‐arginine analogs and flavonoid derivates, such as quercetin pentaacetate, effectively inhibited LPS‐induced NO and PGE2 productions, at the same time, inhibited enhanced expressions of iNOS and COX‐2 genes. J. Cell. Biochem. 82: 537–548, 2001. © 2001 Wiley‐Liss, Inc.
Regulatory role of endogenous regucalcin in the enhancement of nuclear deoxyribonuleic acid synthesis with proliferation of cloned rat hepatoma cells (H4‐II‐E)
Authors: Inagaki Shyuichiroh , Yamaguchi Masayoshi
Published: August 2002

The role of endogenous regucalcin in the regulation of deoxyribonuleic acid (DNA) synthesis in the nuclei of the cloned rat hepatoma cells (H4‐II‐E) with proliferative cells was investigated. Cells were cultured for 6–96 h in a α‐minimum essential medium (α‐MEM) containing fetal bovine serum (FBS; 1 or 10%). Cell number was significantly increased between 24 and 96 h after culture with 10% FBS; cell proliferation was markedly stimulated by culture with 10% FBS as compared with that of 1% FBS. In vitro DNA synthesis activity in the nuclei of cells was significantly elevated 6 h after culture with 10% FBS and its elevation was remarkable at 12 and 24 h after the culture. Nuclear DNA synthesis activity was significantly reduced in the presence of various protein kinase inhibitors (PD98059, staurosprine, or trifluoperazine) in the reaction mixture containing the nuclei of cells cultured for 12 and 24 h with FBS (1 and 10%). The addition of regucalcin (10−7 and 10−6M) in the reaction mixture caused a significant inhibition of nuclear DNA synthesis activity. The presence of anti‐regucalcin monoclonal antibody (25–100 ng/ml) in the reaction mixture containing the nuclei of cells cultured for 24 h with 10% FBS resulted in a significant increase in nuclear DNA synthesis activity. This increase was completely blocked by the addition of regucalcin (10−6 M). The effect of anti‐regucalcin antibody (100 ng/ml) in increasing nuclear DNA synthesis activity was significantly inhibited in the presence of various protein kinase inhibitors. DNA synthesis activity was significantly enhanced in the presence of anti‐regucalcin antibody (100 ng/ml) in the reaction mixture containing the nuclei of cells cultured for 24 h with 10% FBS in the presence of Bay K 8644 (2.5 × 10−6 M). Culture with Bay K 8644 did not cause a significant increase in DNA synthesis activity in the absence of anti‐regucalcin antibody. The present study demonstrates that endogenous regucalcin plays a suppressive role in the enhancement of nuclear DNA synthesis with proliferative cells. J. Cell. Biochem. 82: 704–711, 2001. © 2001 Wiley‐Liss, Inc.
Cloning and characterization of osteoactivin, a novel cDNA expressed in osteoblasts
Authors: Safadi Fayez. F. , Xu Jie , Smock Steven L. , Rico Mario C. , Owen Thomas A. , Popoff Steven N.
Published: August 2002

Osteoblast development is a complex process involving the expression of specific growth factors and regulatory proteins that control cell proliferation, differentiation, and maturation. In this study, we used the rat mutation, osteopetrosis (op), to examine differences in skeletal gene expression between mutant op and normal littermates. Total RNA isolated from long bone and calvaria was used as a template for mRNA differential display. One of many cDNAs that were selectively expressed in either normal or mutant bone was cloned and sequenced and found to share some homology to the human nmb and Pmel 17 genes. This novel cDNA was named osteoactivin. Osteoactivin has an open reading frame of 1716 bp that encodes a protein of 572 amino acids with a predicted molecular weight of 63.8 kD. Protein sequence analysis revealed the presence of a signal peptide and a cleavage site at position 23. The protein also has thirteen predicted N‐linked glycosylation sites and a potential RGD integrin recognition site at position 556. Northern blot analysis confirmed that osteoactivin was 3‐ to 4‐fold overexpressed in op versus normal bone. RT‐PCR analysis showed that osteoactivin is most highly expressed in bone compared with any of the other non‐osseous tissues examined. In situ hybridization analysis of osteoactivin in normal bone revealed that it is primarily expressed in osteoblasts actively engaged in bone matrix production and mineralization. In primary rat osteoblast cultures, osteoactivin showed a temporal pattern of expression being expressed at highest levels during the later stages of matrix maturation and mineralization and correlated with the expression of alkaline phosphatase and osteocalcin. Our findings show that osteoactivin expression in bone is osteoblast‐specific and suggest that it may play an important role in osteoblast differentiation and matrix mineralization. Furthermore, osteoactivin overexpression in op mutant bone may be secondary to the uncoupling of bone resorption and formation resulting in abnormalities in osteoblast gene expression and function. J. Cell. Biochem. 84: 12–26, 2002. © 2001 Wiley‐Liss, Inc.
Inducible expression of the regulatory protein kinase CK2β subunit: Incorporation into complexes with catalytic CK2 subunits and re‐examination of the effects of CK2β on cell proliferation
Authors: Vilk Greg , Derksen D. Richard , Litchfield David W.
Published: August 2002

The regulatory subunit of protein kinase CK2, designated CK2β, exists both free in cells and in complexes with the CK2 catalytic subunits. Growing evidence suggests that CK2β has functions dependent and independent of the CK2 catalytic subunits. There have been indications that CK2β has functions associated with DNA damage responses and in the control of cell proliferation. For example, transient and stable constitutive overexpression of CK2β in mammalian cells was previously shown to perturb cell cycle progression and to attenuate proliferation. To systematically investigate the molecular mechanisms responsible for these effects of CK2β on cell proliferation, we generated human osteosarcoma U2OS cell lines with tetracycline‐regulated expression of CK2β. Increased expression of CK2β results in increases in total cellular CK2 activity, but no changes in cell cycle profiles or proliferation. Furthermore, following exposure to ultraviolet radiation, p53 induction was identical regardless of the levels of CK2β in cells. Mouse 3T3‐L1 cells stably transfected with CK2β also showed no alterations in cell proliferation. The differences between these results and those previously reported emphasize the complex nature of CK2β and its cellular functions. Furthermore, these results indicate that increased expression of CK2β is not by itself sufficient to effect alterations in cell proliferation. J. Cell. Biochem. 84: 84–99, 2002. © 2001 Wiley‐Liss, Inc.
Four‐component relativistic Kohn–Sham theory
Authors: Saue Trond , Helgaker Trygve
Published: August 2002

A four‐component relativistic implementation of Kohn–Sham theory for molecular systems is presented. The implementation is based on a nonredundant exponential parametrization of the Kohn–Sham energy, well suited to studies of molecular static and dynamic properties as well as of total electronic energies. Calculations are presented of the bond lengths and the harmonic and anharmonic vibrational frequencies of Au2, Hg, HgAu+, HgPt, and AuH. All calculations are based on the full four‐component Dirac–Coulomb Hamiltonian, employing nonrelativistic local, gradient‐corrected, and hybrid density functionals. The relevance of the Coulomb and Breit operators for the construction of relativistic functionals is discussed; it is argued that, at the relativistic level of density‐functional theory and in the absence of a vector potential, the neglect of current functionals follows from the neglect of the Breit operator. © 2002 Wiley Periodicals, Inc. J Comput Chem 23: 814–823, 2002
Generate: A program for 3‐D structure generation and conformational analysis of peptides and peptidomimetics
Authors: Bultinck Patrick , Augustynen Stijn , Hilbers Hans W. , Moret Ed E. , Tollenaere Jan P.
Published: August 2002

The program Generate, aimed at generating 3‐D structures for peptides and peptidomimetics, is presented. The algorithm is based on a build‐up procedure, using a library of conformations of amino acid residues. This library is built from conformational analysis of amino acids placed in a di‐ or tripeptide environment to mimic the surroundings of the amino acid in a true peptide, considering different positions of the residue in the peptide chain (peptidyl fragment, NH‐terminus or COO−‐terminus). Cis‐trans isomerism in the amide bonds is taken into account by construction of rotamer libraries for different isomers. Water solvation is included through the GB/SA model. New amino acid residues can easily be added to the libraries, making it possible to generate conformations of peptidomimetics. © 2002 Wiley Periodicals, Inc. J Comput Chem 23: 746–754, 2002
Basis‐set completeness profiles in two dimensions
Authors: Auer Alexander A. , Helgaker Trygve , Klopper Wim
Published: August 2002

A two‐electron basis‐set completeness profile is proposed by analogy with the one‐electron profile introduced by D. P. Chong (Can J Chem 1995, 73, 79). It is defined as Y(α,β)=∑m∑n〈Gα(1)Gβ(2)∣(1/r12)∣Ψm(1)Ψn(2)〉〈Ψm(1)Ψn(2)∣r12∣Gα(1)Gβ(2)〉 and motivated by the expression for the basis‐set truncation correction that occurs in the framework of explicitly correlated methods (Gα is a scanning Gaussian‐type orbital of exponent α and {Ψm} is the orthonormalized one‐electron basis under study). The two‐electron basis‐set profiles provide a visual assessment of the suitability of basis sets to describe electron‐correlation effects. Furthermore, they provide the opportunity to assess the quality of the basis set as a whole—not only of the individual angular momentum subspaces, as is the case for the one‐electron basis‐set profiles. The two‐electron completeness profiles of the cc‐pVXZ (X=D, T, Q), aug‐cc‐pVTZ, cc‐pCVTZ, and SVP‐auxiliary basis sets for the carbon atom are presented as illustrative examples. © 2002 Wiley Periodicals, Inc. J Comput Chem 23: 420–425, 2002
Therapist countertransference: In search of common themes and empirical referents
Authors: Kiesler Donald J.
Published: August 2002

Throughout this issue on countertransference (CT) and elsewhere, the multiple terms used to describe CT events provide rich conceptual diversity, but also result in theoretical isolation and clinical compartmentalization. Our study and understanding of CT would benefit remarkably if we could agree on empirical‐therapist features that any CT conceptualization must target. Subsequent discussions of CT then could specify which of the respective empirical features are being addressed. Accordingly, I offer a descriptive framework that asserts that CT and its hypothesized subtypes (e.g., subjective and objective) can be detected empirically whenever a therapist's experiences and actions with a particular client assume one of several definable “deviations from baseline.” If, in our clinical or research pursuits, we are to understand CT, we have to take the time to assess the necessary behavioral baselines for therapists and their clients. © 2001 John Wiley & Sons, Inc. J Clin Psychol/In Session 57: 1053–1063, 2001.
The Hispanic MMPI‐A across five countries
Authors: Scott Ronald L. , Butcher James N. , Young Tracy L. , Gomez Nelson
Published: August 2002

The Hispanic MMPI‐A was developed for use with Spanish‐speaking adolescents living in the United States. In order to sample its potential usefulness in other countries, the test was administered to 385 Spanish‐speaking adolescents in Colombia, Mexico, Peru, Spain, and the United States. The results showed a high degree of similarity across the five countries on the basic content and supplementary scales. Most scales were within 1/2 standard deviation of the U.S. Hispanic mean, and no scale elevations were greater than T = 65. Slight differences (T = 55–60) were obtained on scale F for males and scales F, Hs, Mf (Peru and Colombia), Pa, and Sc for females. This study would suggest that the Hispanic MMPI‐A, with its established norms, is appropriate for adaptation in Spanish‐speaking countries other than the United States. Further studies with the Hispanic MMPI‐A are recommended to determine its effectiveness in clinical assessment in Spanish‐speaking countries. © 2002 Wiley Periodicals, Inc. J Clin Psychol 58: 407–417, 2002.
Can neighborhood associations be allies in health policy efforts? Political activity among neighborhood associations
Authors: Lenk Kathleen M. , Toomey Traci L. , Wagenaar Alexander C. , Bosma Linda M. , Vessey John
Published: August 2002

We examined organizational characteristics and types of political actions of neighborhood associations, and factors influencing the amount of political activity among the associations. We hypothesized that four neighborhood characteristics (population size, income, educational level, and percentage of owner‐occupied households) and six organizational characteristics (budget, number of staff, size of board, newsletter publication, coalition involvement, and resident involvement) would influence the amount of political activity of the associations. We obtained data from the 1990 U.S. Census and a survey of neighborhood associations (n = 84) in Minneapolis and St. Paul, Minnesota. We found that neighborhood associations engaged in numerous and diverse political activities. Results from multiple regression analyses revealed that median household income was negatively associated with amount of political activity. Population size of the neighborhood and intensity of involvement in multi‐organization coalitions were both positively associated with political activity (all significant at p < .05). We conclude that neighborhood associations, particularly those in larger and poorer neighborhoods, can be key allies in health and social policy efforts. © 2002 John Wiley & Sons, Inc.
Oxygen‐mediated regulation of skeletal muscle satellite cell proliferation and adipogenesis in culture
Authors: Csete Marie , Walikonis Jean , Slawny Nicole , Wei Yuewang , Korsnes Sheryl , Doyle John C. , Wold Barbara
Published: August 2002

Major problems in stem cell biology revolve around defining the developmental potential of cell populations and understanding how their potential is maintained or progressively restricted. Oxygen (O2) is an obvious environmental factor which has received little attention in culturing skeletal muscle progenitor cells. In this work, we examine the effects of O2 levels on the developmental potential, proliferative capacity, and phenotype of the adult skeletal muscle fiber progenitor population (satellite cells), and cell lines that model multipotential embryonic paraxial mesoderm from which skeletal muscle develops. Both satellite cell proliferation and survival of mature fibers increased in physiologic (6%) O2 vs. non‐physiologic 20% O2 used in virtually all traditional cell culture. Six percent O2 conditions also accelerated the up‐regulation of multiple MyoD family myogenic regulatory factors (MRFs). An unexpected finding was that fiber‐adherent satellite cells could assume a non‐myogenic phenotype. By the criteria of molecular markers and gross lipid accumulation, satellite cells were found to assume an adipocyte phenotype, and did so more prominently in 20% O2 than in physiologic O2. Selection of the adipogenic fate and execution of adipogenesis by multipotential mesenchymal cell lines was also dramatically higher in traditional 20 vs. 6% O2, and decreased adipogenesis in physiologic O2 was associated with significantly less expression of the adipogenic regulator, PPARγ. These results suggest that regulatory pathways affected by O2 are important for satellite cell proliferation, execution of cell fate, and parent muscle survival in culture, and so may play a role in vivo under normal or pathologic conditions. © 2001 Wiley‐Liss, Inc.
Malondialdehyde binding to proteins dramatically alters fibroblast functions
Authors: Rittié Laure , Monboisse Jean‐Claude , Gorisse Marie‐Claude , Gillery Philippe
Published: August 2002

The regulation of cell metabolism by the surrounding environment is deeply altered by the posttranslational nonenzymatic modifications of extracellular proteins that occur throughout lifespan in vivo and modify their structural and functional properties. Among them are protein adducts formed by components generated from oxidative processes, such as malondialdehyde (MDA). We have investigated here the effects of MDA‐binding to proteins on cultured fibroblast functions. Type I collagen and/or serum proteins were incubated with 0–100 mM MDA for 3 h before use in fibroblast cultures. In tridimensional lattice cultures, MDA‐treated collagen inhibited the contracting activity of fibroblasts. A similar inhibition of lattice contraction was reproduced by the addition of MDA‐treated serum to the culture medium. In monolayer cultures, the addition of MDA‐modified serum proteins completely inhibited fibroblast multiplication without effect on initial adhesion steps. MDA‐modified proteins decreased the proliferative capacities of cells, strongly altered cell cycle progression by blocking passage to G2/M phases, and induced apoptotic features in fibroblasts. Our results show, for the first time, that MDA‐modified proteins are potentially as deleterious as free MDA, and could be involved in aging as well as in degenerative complications of diseases with increased oxidative stress such as diabetes mellitus or atherosclerosis. J. Cell. Physiol. 191: 227–236, 2002. © 2002 Wiley‐Liss, Inc.
Nitric oxide chemistry and cellular signaling
Authors: Gow Andrew J. , Ischiropoulos Harry
Published: August 2002

Nitric oxide (NO) has been shown to mediate a number of different physiological functions within every major organ system. This wide variety of functional roles is made all the more remarkable when one considers that NO is a simple diatomic molecule. However, despite the simplicity of the molecule, NO possesses a wide range of chemical reactivity and multiple potential reactive targets. It is the variability of NO reactivity, which leads to its capability to control such a vast range of biological functions. In essence the functionality of NO is controlled by its chemical reactivity. In order to understand this possibility further it is necessary to consider the biologically relevant reactions of nitric oxide. © 2001 Wiley‐Liss, Inc.
COP‐1, a member of the CCN family, is a heparin‐induced growth arrest specific gene in vascular smooth muscle cells
Authors: Delmolino Laurie M. , Stearns Nancy A. , Castellot John J.
Published: August 2002

Vascular smooth muscle cell (VSMC) hyperplasia is responsible for the failure of 15–30% of vascular surgical procedures such as coronary artery bypass grafts and angioplasties. We and others have shown that heparin suppresses VSMC proliferation in vivo and in cell culture. We hypothesize that heparin inhibits VSMC proliferation by binding to cell surface receptors, resulting in selective modulation of mitogenic signal transduction pathways and altered transcription of a specific subset of growth regulatory genes. To test this idea, we used subtractive hybridization to identify differentially expressed mRNAs in heparin‐treated and untreated VSMC. We identified a heparin induced mRNA identical to Cop‐1, a member of the CCN family of proteins which are secreted, cysteine‐rich modular proteins involved in growth regulation and migration. Cop‐1 from smooth muscle cells appears to have a different expression pattern and possibly different functions than Cop‐1 from other cells. Cop‐1 mRNA is expressed at high levels in quiescent VSMC and at low levels in proliferating VSMC, an expression pattern highly characteristic of growth arrest specific genes. Cop‐1 mRNA is expressed at high levels in heparin treated VSMC and COP‐1 protein is secreted into culture medium. In tissues, Cop‐1 expression is observed in the uninjured rat aorta suggesting a possible role for Cop‐1 in vivo. We found PDGF, but not EGF, inhibits the expression of Cop‐1 in VSMC. Neither TGF‐β nor interferon‐β, two inhibitors of VSMC proliferation, were able to induce Cop‐1 expression. In addition, heparin does not induce Cop‐1 mRNA in endothelial cells and VSMC resistant to the antiproliferative effect of heparin. Conditioned medium from cells over‐expressing COP‐1 protein inhibits VSMC proliferation in culture. Together, our data indicate that COP‐1 may play a role in the antiproliferative mechanism of action of heparin. © 2001 Wiley‐Liss, Inc.