bluuebunny/crossref_metadata_2025 huggingface dataset:
Breast cancer mortality patterns and time trends in 10 new EU member states: Mortality declining in young women, but still increasing in the elderly
Authors: Tyczynski Jerzy E. , Plesko Ivan , Aareleid Tiiu , Primic‐Zakelj Maja , Dalmas Miriam , Kurtinaitis Juozas , Stengrevics Aivars , Parkin D. Maxwell
Published: August 2004

Breast cancer is the cancer diagnosed most frequently in women worldwide. In Europe it is the most common cancer in the female population, with approximately 350,000 new cases diagnosed each year including 130,000 deaths. Incidence rates are increasing in the majority of European countries, whereas a decline in mortality rates has been observed in many West European countries since the late 1980s and early 1990s. Our study examines breast cancer mortality patterns and time trends in the new European Union (EU) member states and compares them with the situation in current EU member states. A Joinpoint regression analysis was used to assess temporal changes in mortality rates and the trends examined in the light of known risk factors, screening programs and advances in treatment. In the majority of the countries analyzed, a deceleration in the increase of mortality rates appeared, followed by a decrease of mortality in many of them in the second half of the 1990s. The declining tendency was visible primarily in young women, and to a lesser extent in middle‐aged women, whereas in elderly women a continuing increase of mortality was observed. Analysis of mortality data, information from previous publications, as well as analysis of known factors influencing breast cancer risk suggest that changes observed are due mainly to recent advances in treatment rather than changes in lifestyle risk factors or the result of screening programs. Early detection and a shift toward more favorable stage distribution could have played the leading role for mortality decline in younger patients. © 2004 Wiley‐Liss, Inc.
Laboratory markers of tumor burden in nasopharyngeal carcinoma: A comparison of viral load and serologic tests for Epstein‐Barr virus
Authors: Fan Hongxin , Nicholls John , Chua Daniel , Chan K.H. , Sham Jonathan , Lee Shuko , Gulley Margaret L.
Published: August 2004

Epstein‐Barr virus (EBV) is present within the tumor cells of most cases of nasopharyngeal carcinoma (NPC). Recent studies suggest that tumor burden is proportional to the level of EBV DNA in blood and that rapid blood testing can be used to guide therapeutic intervention. The relative utility of viral load vs. serology has been insufficiently studied. In our study, EBV viral load was measured by quantitative PCR using either real‐time or end‐point detection systems in serum samples from 124 NPC patients (93 pretreatment, 13 relapsed, 18 in remission) and 40 controls. Serologic titers against EBV early antigen were measured in the same serum samples. EBV DNA was detectable in 64 of 93 untreated NPC patients (69%; mean viral load 11,211 copies/ml), 11 of 13 relapsed NPC patients (85%; mean 53,039 copies/ml) and 0 of 18 remission patients. EBV DNA was detectable in only 1 of 40 non‐NPC controls (3%). In 34 instances where paired plasma and serum samples were available for testing, both were effective sample types, and there was no significant difference between end‐point and real‐time methods for measuring viral load. Early antigen (EA) IgA and IgG titers were elevated in most NPC patients regardless of whether their disease was active or in remission. EBV viral load was more informative than was EA serology for distinguishing remission from relapsed disease. EBV DNA measurement appears to be a noninvasive way to monitor tumor burden after therapy. © 2004 Wiley‐Liss, Inc.
Long‐term activation of SAPK/JNK, p38 kinase and fas‐L expression by cisplatin is attenuated in human carcinoma cells that acquired drug resistance
Authors: Brozovic Anamaria , Fritz Gerhard , Christmann Markus , Zisowsky Jochen , Jaehde Ulrich , Osmak Maja , Kaina Bernd
Published: August 2004

Tumor cells chronically exposed to cisplatin (cDDP) acquire cDDP resistance that impacts tumor therapy. To elucidate the mechanism of acquired cDDP resistance (ACR), we compared HeLa cells that gained ACR upon chronic cDDP treatment with the parental strain. We show that ACR is due to a lower level of induced apoptosis. Further, upon cDDP treatment, the levels of Fas, Bax and Bid remained unchanged, whereas Bcl‐2 and p‐Bad were reduced at late times (120 hr) after treatment. At early times, Fas ligand (fas‐L) expression was significantly enhanced in sensitive compared to resistant cells and remained upregulated up to the onset of apoptosis. Thus, activation of the Fas system is critical, which is in line with the finding that in sensitive cells, caspase‐8 along with caspase‐9 and ‐3 were activated by cDDP. cDDP provoked the activation of stress‐activated protein kinase/c‐Jun N‐terminal kinase (SAPK/JNK) and p38 kinase dose‐dependently, with significantly lower levels in ACR cells than in the sensitive parental line. cDDP induces c‐Jun and AP‐1 activity, as measured by a reporter gene assay, which was again attenuated in ACR cells. Time course analysis revealed that SAPK/JNK and p38 kinase activity was sustained upregulated (> 72 hr postexposure), which occurred at much higher level in sensitive than in ACR cells. Inhibition of either JNK or p38 kinase (by JNK inhibitor II and SB 203580, respectively) attenuated cDDP‐induced apoptosis, supporting the role of JNK and p38 kinase in the cDDP response. Since several independently derived cDDP‐resistant cell lines displayed attenuated MAPK signaling, sustained SAPK/JNK and p38 kinase activation may be a general mechanism of cDDP‐induced cell death. ACR cells displayed a reduced level of DNA damage, indicating long‐term stimulation of SAPK/JNK and p38 kinase is triggered by nonrepaired cDDP‐induced DNA lesions. © 2004 Wiley‐Liss, Inc.
JTE‐522, a selective COX‐2 inhibitor, interferes with the growth of lung metastases from colorectal cancer in rats due to inhibition of neovascularization: A vascular cast model study
Authors: Kobayashi Hirotoshi , Gonda Tsuyoshi , Uetake Hiroyuki , Higuchi Tetsuro , Enomoto Masayuki , Sugihara Kenichi
Published: August 2004

The lung is a frequent site of metastasis from colorectal cancer, but angiogenesis of lung metastases has not been clarified. Some COX‐2 inhibitors prevent tumor growth, although the inhibitory mechanism at the metastatic site is obscure. We investigated the microvascular structure of small lung metastases and the effect of JTE‐522, a selective COX‐2 inhibitor, on the angiogenesis of pulmonary metastases from colorectal cancer in rats. The tail veins of 25 male F344/DuCrj rats, aged 5 weeks, were injected with a tumor suspension containing 5 × 106 RCN‐9, a rat colon cancer cell line. Three weeks later, pulmonary vascular resin corrosion casts were taken and the vascularity of metastases was studied using stereo and scanning electron microscopes. We investigated the effect of 0, 10 and 30 mg/kg/day of JTE‐522 on the angiogenesis of pulmonary metastases in 3 groups of 5 male rats out of 25. JTE‐522 reduced the diameter of tumor vessels as well as the number and size of metastatic tumors. The diameter of tumor vessels and the size of lung metastases significantly and positively correlated with neovascularization in the control group, but not in the JTE‐522‐treated groups. JTE‐522 also affected type of vasculature of metastases, which differed depending on their size. JTE‐522 interfered with the growth of hematogenous metastatic tumors by disrupting neovascularization. However, JTE‐522 may have some important mechanisms other than inhibition of neovascularization. JTE‐522 may be one of the therapeutic agents for the treatment of hematogenous metastasis of colorectal cancer. © 2004 Wiley‐Liss, Inc.
Expression of the nucleoside‐derived drug transporters hCNT1, hENT1 and hENT2 in gynecologic tumors
Authors: Farré Xavier , Guillén‐Gómez Elena , Sánchez Lydia , Hardisson David , Plaza Yolanda , Lloberas Jorge , Casado F. Javier , Palacios José , Pastor‐Anglada Marçal
Published: August 2004

Deoxynucleoside analogs are used in the treatment of a variety of solid tumors. Their transport across the plasma membrane may determine their cytotoxicity and thus nucleoside transporter (NT) expression patterns may be of clinical relevance. Lack of appropriate antibodies for use in paraffin‐embedded biopsies has been a bottleneck to undertake high‐throughput analysis of NT expression in solid tumors. Here we report the characterization of 2 new antibodies raised against the low‐affinity equilibrative NTs, hENT1 and hENT2, suitable for that purpose. These 2 antisera, along with a previously characterized antibody that specifically recognizes the high‐affinity Na‐dependent concentrative NT, hCNT1, have been used to analyze, using a tissue array approach, NT expression in gynecologic cancers (90 ovarian, 80 endometrial and 118 uterine cervix carcinomas). Human CNT1 was not detected in 33% and 39% of the ovarian and uterine cervix carcinomas, respectively, whereas hENT1 and hENT2 expression was significantly retained in a high percentage of tumors (91% and 96% for hENT1, 84% and 98% for hENT2, in ovarian and cervix carcinomas, respectively). Only a few endometrial carcinomas (15%) were found to be negative for hCNT1, but they all retained hENT1 and hENT2 expression. In ovarian cancer, the loss of all 3 NT proteins was a more common event in the clear cell histologic subtype than in the serous, mucinous and endometrioid histotypes. In uterine cervix tumors, the loss of expression of hCNT1 was significantly associated with the adenocarcinoma subtype. In summary, hCNT1 was by far the isoform whose expression was most frequently reduced or lost in the 3 types of gynecologic tumors analyzed. Moreover, NT expression is related to the type of gynecologic tumor and its specific subtype, hCNT1 protein loss being highly correlated with poor prognosis histotypes. Since hCNT1, hENT1 and hENT2 recognize fluoropyrimidines as substrates, but with different affinities, this study anticipates high variability in drug uptake efficiency in solid tumors. © 2004 Wiley‐Liss, Inc.
Downregulation of tissue factor by RNA interference in human melanoma LOX‐L cells reduces pulmonary metastasis in nude mice
Authors: Wang Xiaolin , Wang Mengyu , Amarzguioui Mohammed , Liu Fang , Fodstad Øystein , Prydz Hans
Published: August 2004

Tissue factor (TF) is the membrane receptor of the serine protease coagulation factor VIIa (FVIIa). Formation of the TF/FVIIa complex initiates the coagulation cascade. We used short hairpin RNA (shRNA)‐mediated RNA interference to knock down TF expression in the human metastatic melanoma cell line LOX‐L. After transfection with the shRNA construct, 3 stable clones with significantly downregulated TF expression were established. They exhibited decreased proliferation in vitro as determined by 14C thymidine incorporation and soft agar assay. The in vivo metastatic potential was assessed in an experimental pulmonary metastasis model in which cells from different clones were injected into the tail vein of nude mice. The incidence of pulmonary tumors was significantly lower in mice receiving shRNA‐expressing cells (33% ± 15%) than in control mice injected with wild‐type cells or cells stably transfected with empty expression vector (90% ± 10%). The mice injected with TF‐downregulated cells had markedly longer survival time (69 ± 17 days) compared to the control mice (35.6 ± 5 days; p = 0.03). Thus, reduction of TF levels in LOX‐L cells significantly delayed and reduced lung tumor formation. As a first step in elucidating the molecular basis for this effect, we compared the global gene expression profile in TF‐downregulated cells and control cells by using cDNA microarray analysis. Forty‐four known human genes were found to be significantly upregulated (> 2‐fold; p < 0.05) and 228 genes significantly downregulated (≥ 3‐fold; p < 0.05) in TF‐downregulated cells compared to control cells. The differentially expressed genes encode proteins functioning in transcription, translation, cell communication and cell growth/death. The results provide a basis for investigating molecular mechanisms underlying the effects of TF on the metastatic capacity of LOX‐L melanoma cells. © 2004 Wiley‐Liss, Inc.
Efficient use of nonequilibrium measurement to estimate free energy differences for molecular systems
Authors: Ytreberg F. Marty , Zuckerman Daniel M.
Published: August 2004

A promising method for calculating free energy differences ΔF is to generate nonequilibrium data via “fast‐growth” simulations or by experiments—and then use Jarzynski's equality. However, a difficulty with using Jarzynski's equality is that ΔF estimates converge very slowly and unreliably due to the nonlinear nature of the calculation—thus requiring large, costly data sets. The purpose of the work presented here is to determine the best estimate for ΔF given a (finite) set of work values previously generated by simulation or experiment. Exploiting statistical properties of Jarzynski's equality, we present two fully automated analyses of nonequilibrium data from a toy model, and various simulated molecular systems. Both schemes remove at least several kBT of bias from ΔF estimates, compared to direct application of Jarzynski's equality, for modest sized data sets (100 work values), in all tested systems. Results from one of the new methods suggest that good estimates of ΔF can be obtained using 5–40‐fold less data than was previously possible. Extending previous work, the new results exploit the systematic behavior of bias due to finite sample size. A key innovation is better use of the more statistically reliable information available from the raw data. © 2004 Wiley Periodicals, Inc. J Comput Chem 25: 1749–1759, 2004
Immunophenotypic analysis of human articular chondrocytes: Changes in surface markers associated with cell expansion in monolayer culture
Authors: Diaz‐Romero Jose , Gaillard Jean Philippe , Grogan Shawn Patrick , Nesic Dobrila , Trub Thomas , Mainil‐Varlet Pierre
Published: August 2004

Cartilage tissue engineering relies on in vitro expansion of primary chondrocytes. Monolayer is the chosen culture model for chondrocyte expansion because in this system the proliferative capacity of chondrocytes is substantially higher compared to non‐adherent systems. However, human articular chondrocytes (HACs) cultured as monolayers undergo changes in phenotype and gene expression known as “dedifferentiation.” To gain a better understanding of the cellular mechanisms involved in the dedifferentiation process, our research focused on the characterization of the surface molecule phenotype of HACs in monolayer culture. Adult HACs were isolated by enzymatic digestion of cartilage samples obtained post‐mortem. HACs cultured in monolayer for different time periods were analyzed by flow cytometry for the expression of cell surface markers with a panel of 52 antibodies. Our results show that HACs express surface molecules belonging to different categories: integrins and other adhesion molecules (CD49a, CD49b, CD49c, CD49e, CD49f, CD51/61, CD54, CD106, CD166, CD58, CD44), tetraspanins (CD9, CD63, CD81, CD82, CD151), receptors (CD105, CD119, CD130, CD140a, CD221, CD95, CD120a, CD71, CD14), ectoenzymes (CD10, CD26), and other surface molecules (CD90, CD99). Moreover, differential expression of certain markers in monolayer culture was identified. Up‐regulation of markers on HACs regarded as distinctive for mesenchymal stem cells (CD10, CD90, CD105, CD166) during monolayer culture suggested that dedifferentiation leads to reversion to a primitive phenotype. This study contributes to the definition of HAC phenotype, and provides new potential markers to characterize chondrocyte differentiation stage in the context of tissue engineering applications. © 2004 Wiley‐Liss, Inc.
Star coloring of graphs
Authors: Fertin Guillaume , Raspaud André , Reed Bruce
Published: August 2004

A star coloring of an undirected graph G is a proper vertex coloring of G (i.e., no two neighbors are assigned the same color) such that any path of length 3 in G is not bicolored. The star chromatic number of an undirected graph G, denoted by χs(G), is the smallest integer k for which G admits a star coloring with k colors. In this paper, we give the exact value of the star chromatic number of different families of graphs such as trees, cycles, complete bipartite graphs, outerplanar graphs, and 2‐dimensional grids. We also study and give bounds for the star chromatic number of other families of graphs, such as planar graphs, hypercubes, d‐dimensional grids (d ≥ 3), d‐dimensional tori (d ≥ 2), graphs with bounded treewidth, and cubic graphs. We end this study by two asymptotic results, where we prove that, when d tends to infinity, (i) there exist graphs G of maximum degree d such that $\chi _s(G) = \Omega({d^{{3\over 2}}\backslash ({\rm log}\ {d})^{1\over 2}})$ and (ii) for any graph G of maximum degree d, $\chi _s(G) = O({d^{{3\over 2}}})$. © 2004 Wiley Periodicals, Inc. J Graph Theory 47: 163–182, 2004
Biomechanical modeling and sensitivity analysis of bipedal running ability. I. Extant taxa
Authors: Hutchinson John R.
Published: August 2004

I used a simple mathematical model of the inverse dynamics of locomotion to estimate the minimum muscle masses required to maintain quasi‐static equilibrium about the four main limb joints at mid‐stance of fast running. Models of 10 extant taxa (a human, a kangaroo, two lizards, an alligator, and five birds) were analyzed in various bipedal poses to examine how anatomy, size, limb orientation, and other model parameters influence running ability. I examined how the muscle masses required for fast running compare to the muscle masses that are actually able to exert moments about the hip, knee, ankle, and toe joints, to see how support ability varies across the limb. I discuss the assumptions and limitations of the models, using sensitivity analysis to see how widely the results differed with feasible parameter input values. Even with a wide range of input values, the models validated the analysis procedure. Animals that are known to run bipedally were calculated as able to preserve quasi‐static equilibrium about their hindlimb joints at mid‐stance, whereas non‐bipedal runners (iguanas and alligators) were recognized as having too little muscle mass to run quickly in bipedal poses. Thus, this modeling approach should be reliable for reconstructing running ability in extinct bipeds such as nonavian dinosaurs. The models also elucidated how key features are important for bipedal running capacity, such as limb orientation, muscle moment arms, muscle fascicle lengths, and body size. None of the animals modeled had extensor muscle masses acting about any one joint that were 7% or more of their body mass, which provides a reasonable limit for how much muscle mass is normally apportioned within a limb to act about a particular joint. The models consistently showed that a key biomechanical limit on running ability is the capacity of ankle extensors to generate sufficiently large joint moments. Additionally, the analysis reveals how large ratite birds remain excellent runners despite their larger size; they have apomorphically large extensor muscles with relatively high effective mechanical advantage. Finally, I reconstructed the evolution of running ability in the clade Reptilia, showing that the ancestors of extant birds likely were quite capable runners, even though they had already reduced key hip extensors such as M. caudofemoralis longus. J. Morphol. 262:421–440, 2004. © 2004 Wiley‐Liss, Inc.
Functional morphology of the reproductive system of Galathea intermedia (Decapoda: Anomura)
Authors: Kronenberger Katrin , Brandis Dirk , Türkay Michael , Storch Volker
Published: August 2004

Spermatophore formation in Galathea intermedia begins in the proximal part of the vas deferens. The contents subsequently form a spermatophoric ribbon, the so‐called “secondary spermatophore,” in its distal part. A strongly muscular ductus ejaculatorius is present in the coxa of the fifth pereiopod which builds up pressure for the extrusion of the spermatophoric ribbon. After extrusion, the ribbon is caught by the first gonopod, while the second gonopod dissolves the matrix of the ribbon. During copulation the spermatophores are randomly placed on the sternum of the female, near the genital opening, by the fifth pereiopods of the male. Subsequent ovulation of the female via the genital opening, an active process accomplished through muscular activity, results in fertilization of the eggs by the exploding spermatophores. External intersexes are characterized by both male and female external sexual characters, but in all individuals only male gonads are present. No trace of a female reproductive system could be detected. Thus, these external intersexes are exclusively functional males. J. Morphol. 262:500–516, 2004. © 2004 Wiley‐Liss, Inc.
Prevalence of gait disorders in hospitalized neurological patients
Authors: Stolze Henning , Klebe Stephan , Baecker Christoph , Zechlin Christiane , Friege Lars , Pohle Sabine , Deuschl Günther
Published: August 2004

The prevalence of gait disorders among neurological inpatients is unknown, although disturbed gait is a common symptom. Gait disorders often lead to loss of independence with restraints for the patients and caregivers and costs for the health system. We designed a prospective study and investigated all patients admitted to a neurological hospital during a 100‐day period for the presence of a gait disorder. Clinical investigation and several disease‐specific rating scales were carried out for 493 patients. In 60% of the patients, a disturbance of gait was diagnosed. Most frequent diagnoses were stroke (21%), Parkinson's disease (17%), and polyneuropathy (7%). Within these diagnoses, the rate of patients with disturbed gait was high in Parkinson's disease (93%), subcortical arteriosclerotic encephalopathy (85%), and motor neuron disease (83%). Advanced age, dementia, alcohol abuse, and treatment with antiepileptics, neuroleptics, benzodiazepines, and chemotherapeutics were identified as risk factors for a gait disorder. A decline of cognitive function was accompanied by a reduction of walking speed. According to these results, gait disorders are among the most frequent symptoms in neurology. © 2004 Movement Disorder Society
A GIAO/DFT study of 1H, 13C and 15N shieldings in amines and its relevance in conformational analysis
Authors: Alkorta Ibon , Elguero José
Published: August 2004

The 1H, 13C and 15N absolute shieldings of 13 amines were calculated at the GIAO/B3LYP/6–311++G** level. For some compounds (ethylamine, piperidine and 1‐methylpiperidine) two conformations were calculated. The 13C and 15N data could be correctly correlated with experimental chemical shifts, allowing the conformation of 1‐methylpiperidine to be established. The 1H NMR absolute shieldings, although less well correlated with δ values, were used to account for the anisotropy effects of the N lone pair. Copyright © 2004 John Wiley & Sons, Ltd.
Motor conduction studies for prognostic assessment of obstetrical plexopathy
Authors: Heise Carlos O. , Lorenzetti Lucilia , Marchese Antonio J. T. , Gherpelli José Luiz D.
Published: August 2004

Early prognostic assessment of obstetrical brachial plexopathies (OBP) would facilitate rational selection of infants for brachial plexus surgery. We performed bilateral motor nerve conduction studies (MNCS) of axillary, musculocutaneous, radial, median, and ulnar nerves in 33 babies (age 10–60 days) with OBP in order to compare the amplitude of compound muscle action potentials (CMAPs). All babies were followed up until 6 months of age and the outcome was classified according to muscle strength and arm function. A CMAP amplitude reduction of more than 90%, compared to the unaffected side, predicted severe weakness of the corresponding root level (p < 0.01). Our results indicate that MNCS are a useful tool for very early prognostic assessment of OBP. Muscle Nerve 30: 451–455, 2004
Effects of exercise and steroid on skeletal muscle apoptosis in the mdx mouse
Authors: Lim Jeong‐Hoon , Kim Dai‐Youl , Bang Moon Suk
Published: August 2004

Reports concerning the influence of exercise loading and steroid administration on dystrophinopathy are inconsistent. To investigate the effect of muscle exercise in Duchenne muscular dystrophy (DMD), 15 control and 15 mdx mice, an animal model of DMD, were divided into free‐living (n = 6), exercise (n = 6), and immobilization (n = 3) groups. Free‐living and exercise groups were further divided into steroid‐treated and sham‐treated groups to evaluate the effect of steroid administration. We measured apoptotic changes by in situ DNA nick‐end labeling (TUNEL), DNA fragmentation assay, and Western blotting for Bcl‐2 and BAX. Apoptosis was most prominent in the sham‐treated exercise group, and it was significantly reduced in the steroid‐treated exercise group. The steroid‐treated free‐living group showed a higher rate of apoptotic change than the sham‐treated free‐living group. Apoptosis was minimized in the free‐living condition, whereas exercise loading and immobilization caused apoptotic change in this muscular dystrophy animal model. Steroid administration induced apoptosis in muscle of free‐living mice, but alleviated the apoptotic damage caused by exercise loading in mdx mice. These findings suggest that steroid administration may be effective in preventing a postexercise deterioration of skeletal muscle in animal models of DMD. Muscle Nerve 30: 456–462 2004
Collagen synthesis and degradation in polyneuropathy and myopathies
Authors: Ahtikoski Anne M. , Tuominen Hannu , Korpelainen Juha T. , Takala Timo E. S. , Oikarinen Aarne
Published: August 2004

To determine the mechanisms underlying the changes in collagen metabolism responsible for muscle fibrosis in patients with neuromuscular diseases, the synthesis and degradation of collagens was studied in muscles of patients with polyneuropathy and noninflammatory myopathies. The mRNA levels for type I, III, and IV collagens and immunohistochemical staining intensities for collagen propeptides and telopeptides were increased in polyneuropathy, suggesting enhanced synthesis of collagens. In myopathy, the mRNA levels were at the control level. Matrix metalloproteinase (MMP)‐2 mRNA level was increased in polyneuropathy, although the quantity of proMMP‐2 was not changed. An increase in type IV collagen concentration and proMMP‐9 expression was observed in polyneuropathy but not in myopathies. Our results suggest that intramuscular accumulation of type IV collagen occurs in polyneuropathy, possibly leading to thickening of the capillary and muscle fiber basement membranes. This may complicate the transportation of nutrients and cellular excreta between blood and muscle cells. Muscle Nerve, 2004
A universal method for structural static reanalysis of topological modifications
Authors: Chen Su Huan , Yang Zhi Jun
Published: August 2004

This paper presents a universal method, iterative combined approximation (iterative CA) approach, for structural static reanalysis of all types of topological modifications. The proposed procedure is basically an approximate two‐step method. First, the newly added degrees of freedom (DOFs) are assumed to be linked to the original DOFs of the modified structure by means of the Guyan reduction so as to obtain the condensed equation. Second, the displacements of the original DOFs of the modified structure are solved by using the iterative CA approach. And the displacements of the newly added DOFs resulting from topological modification can be recovered. Four numerical examples are given to illustrate the applications of the present approach. The results show that the proposed method is effective for structural static reanalysis of all types of the topological modifications and it is easy to implement on a computer. Copyright © 2004 John Wiley & Sons, Ltd.
Polyelectrolyte spin assembly: Influence of ionic strength on the growth of multilayered thin films
Authors: Lefaux Christophe J. , Zimberlin Jessica A. , Dobrynin Andrey V. , Mather Patrick T.
Published: August 2004

Layer‐by‐layer (LbL) assembly of polymer electrolyte multilayers is now a well‐established method for the fabrication of thin films by sequential adsorption of alternating layers of oppositely charged polyelectrolytes. Most commonly, such adsorptions have been from quiescent solutions of varying ionic strength and pH. Here, we report results on an alternative processing route for the achievement of polymeric multilayer assemblies of poly(sodium‐4‐styrene sulfonate) and poly(allylamine hydrochloride) that utilizes conventional spin coating. We investigated and describe herein the dependence of multilayer film buildup on solution ionic strength for comparison with similar dependence in quiescent adsorption. Using UV‐Vis spectroscopy we monitored the growth of the multilayered films, while with Atomic Force Microscopy (AFM) we examined the surface features and measured coating thicknesses at different salt concentrations. AFM and UV‐Vis data reveal two regimes of behavior with increasing salt: strong salt‐dependence at low salt contents, and weak salt‐dependence for high salt contents. To explain this observation, we introduce the relevance of the dimensionless groupDe=τ, the local Deborah Number, to the problem. As ionic strength increases, τ increases so that spin‐assembly flow influences adsorbed conformation, and thus LbL growth rate. Our results indicate the ability to design and control polyelectrolyte multilayered structures prepared via spin assembly by varying solution properties that influence the conformation of deposited polymer chains. Additionally, our studies reveal the need for study of the fundamental mechanisms of polyelectrolyte adsorption within complex flow fields. © 2004 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 42: 3654–3666, 2004
Engineering of a polymeric bacterial protein as a scaffold for the multiple display of peptides
Authors: Laplagne D.A. , Zylberman V. , Ainciart N. , Steward M. W. , Sciutto E. , Fossati C.A. , Goldbaum F. A.
Published: August 2004

Protein assemblies with a high degree of repetitiveness and organization are known to induce strong immune responses. For that reason they have been postulated for the design of subunit vaccines by means of protein engineering. The enzyme lumazine synthase from Brucella spp. (BLS) is highly immunogenic, presumably owing to its homodecameric arrangement and remarkable thermodynamic stability. Structural analysis has shown that it is possible to insert foreign peptides at the ten amino terminus of BLS without disrupting its general folding. These peptides would be displayed to the immune system in a highly symmetric three‐dimensional array. In the present work, BLS has been used as a protein carrier of foreign peptides. We have established a modular system to produce chimeric proteins decorated with ten copies of a desired peptide as long as 27 residues and have shown that their folding and stability is similar to that of the wild‐type protein. The knowledge about the mechanisms of dissociation and unfolding of BLS allowed the engineering of polyvalent chimeras displaying different predefined peptides on the same molecular scaffold. Moreover, the reassembly of mixtures of chimeras at different steps of the unfolding process was used to control the stoichiometry and spatial arrangement for the simultaneous display of different peptides on BLS. This strategy would be useful for vaccine development and other biomedical applications. Proteins 2004. © 2004 Wiley‐Liss, Inc.
Density matrix force‐balance equation applied to He, Be, and Ne atoms and to almost‐spherical methane‐like molecules
Authors: Howard I. A. , March N. H. , Bartha F. , Ray A. K. , Zhang M.‐L.
Published: August 2004

Holas and March (Phys Rev A 1995, 51, 2040) expressed the force −∇Vxc(r) associated with the exchange correlation potential Vxc(r) of density functional theory in terms of low‐order density matrices. Their result utilizes a many‐electron force balance equation for −∇Vext(r), where Vext(r) denotes the external potential. Here, insight is sought into this force‐balance equation by using both Hartree–Fock results and correlated electron calculations. Examples of two‐electron systems, including the exactly solvable model Hookean atom, are first set out, followed by treatments of the Be and Ne atoms. As a final, molecular, example, methane‐like systems are also more briefly studied, but now restricted solely to the Hartree–Fock level of approximation. © 2003 Wiley Periodicals, Inc. Int J Quantum Chem, 2004
Overlap integrals for Dirac–Slater orbitals
Authors: Talman James D.
Published: August 2004

The problem of computing overlap integrals of Slater‐type orbitals on different centers for relativistic orbitals with a behavior of rs at the nuclei is considered. The integrals can be expressed as integrals in momentum space involving a spherical Bessel function jL(kR). The problem of the oscillatory behavior of jL(kR) at large kR can be eliminated by transforming the integral to a contour integral in the upper half‐plane. A method of carrying out the numerical integration is described and the number of integration points required for a large number of cases are given. © 2003 Wiley Periodicals, Inc. Int J Quantum Chem, 2004
Modulation of bradykinin‐induced calcium changes in SH‐SY5Y cells by neurosteroids and sigma receptor ligands via a shared mechanism
Authors: Hong Weimin , Nuwayhid Samer J. , Werling Linda L.
Published: August 2004

In this study we investigated the effects of sigma receptor ligands and neurosteroids on bradykinin‐induced intracellular calcium concentration ([Ca2+]i) changes in SH‐SY5Y neuroblastoma cells. [Ca2+]i levels in cells loaded with fura‐2 were monitored with dual‐wavelength ratiometric fluorescence measurement. Submicromolar concentrations of bradykinin elicited [Ca2+]i responses with a fast rise followed by a slow decline in these cells. Preincubation of low micromolar concentrations of the neurosteroids pregnenolone, dehydroepiandrosterone (DHEA), or the prototypic sigma (σ) receptor agonist (+)pentazocine potentiated bradykinin‐induced [Ca2+]i changes in SH‐SY5Y cells. The σ receptor antagonist haloperidol blocked the enhancing effects on [Ca2+]i by (+)pentazocine or pregnenolone. Progesterone did not significantly affect the basal [Ca2+]i level or bradykinin‐induced [Ca2+]i changes in these cells. However, coincubation of progesterone with (+)pentazocine, pregnenolone, or DHEA reversed their potentiating effects. The antagonistic effects of haloperidol and progesterone on the potentiating effects of (+)pentazocine and pregnenolone suggested that these ligands might act through a common mechanism. We further showed that progesterone, pregnenolone, and DHEA competed for [3H](+)pentazocine binding in SH‐SY5Y cells with Ki values of 0.13 ± 0.03 μM, 0.98 ± 0.34 μM, and 5.2 ± 1.4 μM, respectively. Thus, the modulation of bradykinin‐induced [Ca2+]i changes by neurosteroids in these cells is likely due to their actions on σ receptors. Synapse 54:102–110, 2004. © 2004 Wiley‐Liss, Inc.
New international business perspectives on Pakistan
Authors: Khan Omar J. , Amine Lyn S.
Published: August 2004

What image comes to mind when one thinks of Pakistan? Prior to the attacks of September 11, 2001, and the subsequent invasion of Afghanistan, many businesspeople had little information about this ancient land. Recently, Pakistan has come to the world's attention as a powerful ally in the U.S.‐led war against terrorism. Situated at the crossroads of the Middle East, Southeast Asia, China, and member nations of the former Soviet Union, Pakistan has long been a leading player in regional political and economic activity. This article presents some new perspectives on Pakistan as a market worthy of a closer look. We survey Pakistan's history and geography, as well as its sociocultural, political‐legal, economic, and competitive environments with a view to identifying new opportunities for foreign investors and global marketers. Current business trends and marketing opportunities are discussed, concluding with an outlook for future growth and development. Valuable insights are offered to businesspeople interested in doing business in Pakistan, a little‐studied but vitally important newly industrializing country (NIC) and big emerging market (BEM). © 2004 Wiley Periodicals, Inc.
Problems and challenges facing the islamic banking system in the west: The case of the UK
Authors: Karbhari Yusuf , Naser Kamal , Shahin Zerrin
Published: August 2004

This study investigates the main problems, challenges, and opportunities facing Islamic banking in the United Kingdom. The study reports the results of interviews that were undertaken with senior officials of several key financial institutions who have had many years of experience in dealing with Islamic banking. Our interviews revealed that, although by strict definition Islamic banks do not currently exist in the UK, London is one of the major centers for Islamic banking and finance. It is apparent that the experience of previously established Islamic institutions, such as Al‐Baraka, has made other institutions realize that it is possible to provide Islamic banking services in the UK under nonbanking regulations. The interviews also revealed that the main problem that Islamic banking faces in the UK is heterogeneous clients and potential clients. Moreover, regulatory hurdles, competition from conventional banks, and lack of adequately qualified and trained personnel exacerbate the situation. The study concludes by identifying opportunities such as e‐banking that may have a significant impact on the future of Islamic banking in the UK. © 2004 Wiley Periodicals, Inc.
An analysis of U.S. small and medium‐sized manufacturers' international business relationships
Authors: Leonidou Leonidas C. , Kaminarides John S. , Hadjimarcou John
Published: August 2004

Notwithstanding the dominant role of the United States in world trade, little is known about the relationships between smaller indigenous manufacturers and their foreign customers and how these differ from domestic market relationships. Using data from a recent study among 201 U.S. small and medium‐sized enterprises (SMEs), the article reveals that (a) these firms tend to put much more emphasis on the domestic side of their business, while the foreign market receives secondary treatment; (b) the atmosphere governing their relational exchanges differs in the two types of markets, being relatively healthier in domestic business; and (c) their domestic business performance levels, although moderate, are significantly higher than those found in foreign markets. © 2004 Wiley Periodicals, Inc.
Global acquisitions, strategic integration and the human factor
Authors: Hunt John W.
Published: August 2004

Stan Lees. 2003. Global Acquisitions, Strategic Integration and the Human Factor. Houndmills, Basingstoke, UK: Palgrave Macmillan. ISBN: 0‐333‐77629‐1
MAP kinase kinase: A node connecting multiple pathways
Authors: Mordret Guy
Published: August 2004

Summry— Numerous studies have been published these last few years on the involvement of MAP kinases in signal transduction reflecting their importance in cell cycle and cell growth controls. The identification and the characterization of their direct upstream activator has considerably enlarged our understanding of the phosphorylation network. The MAP kinase kinases (MAPKKs) are dual‐specificity protein kinases which phosphorylate and activate MAP kinases. To date, MAPKK homologues have been found in yeast, invertebrates, amphibians, and mammals. Moreover, the MAPKK/MAPK phosphorylation switch constitutes a basic module activated in distinct pathways in yeast and in vertebrates. MAPKK regulation studies have led to the discovery of at least four MAPKK convergent pathways in higher organisms. One of these is similar to the yeast pheromone response pathway which includes the ste11 protein kinase. Two other pathways require the activation of either one or both of the serine/threonine kinase‐encoded oncogenes c‐Raf‐I and c‐Mos. Additionally, recent studies suggest a possible effect of the cell cycle control regulatory cyclin‐dependent kinase 1 (cdc2) on MAPKK activity. Finally, MAPKKs seem to be essential transducers through which signals must pass before reaching the nucleus.
A SV‐40 immortalized murine endothelial cell line from peripheral lymph node high endothelium expresses a new α‐L‐fucose binding protein
Authors: Bizourne Nadine , Denis Véronique , Legrand Alain , Monsigny Michel , Kieda Claudine
Published: August 2004

Summry— Endothelial cells from mouse peripheral lymph nodes were immortalized by cationic liposome‐mediate transfection using a plasmid construct containing both the gene coding for the large T antigen of simian virus 40 and a geneticin resistance gene suitable for selection. A cell line (HECal10) was isolated on the basis of its capacity to specifically bind fucoside carrying glycoconjugates; these cells present the main characteristics of endothelial cells: production of angiotensin converting enzyme and of factor VII‐related antigen. Upon stimulation, they express E‐selectin which binds oligosaccharides containing the Lewisx determinant (Fucα3[Galβ4 GlcNacβ3Galβ) and the MECA 79 addressin which is characteristics for the peripheral lymph node high endothelium and is a L‐selectin. HECa10 cells, as well as peripheral lymph node high endothelial cells in primary culture, express a second fucoside binding protein which differs from E‐selectin. Indeed, this new fucoside‐binding protein is constitutively expressed on unstimulated cells while E‐selectin is not. Furthermore, HECa10 cells mediate selective lymphoid cells adhesion in a selectin/addressin‐dependent mechanism, mainly inhibited by MECA 79 antibody and, in a fucose‐binding lectin‐dependent manner, mainly inhibited by the specifc neoglycoprotein.
Endocytosis and intracellular degradation of heterologous protein by eosinophilic granulocytes isolated from rainbow trout (Oncorhynchus mykiss) posterior intestine
Authors: Dorin Dominique , Sire Marie‐France , Vernier Jean‐Marie
Published: August 2004

Summry— Adherence capacity to tissue substrate, endocytosis capacity for heterologous proteins, and proteolytic activity were determined in intestinal granulocytes (EGCs) isolated from healthy adult rainbow trout. The percentage of cells that could adhere to a smooth plastic surface increased with increasing incubation time. Endocytosis was effective for heterologous (human immunoglobulin G, IgGh; ovine somatotropin, oST) but not homologous proteins (recombinant trout somatotropin, rtST). The activity of cathepsin D increased significantly after the endocytosis of a heterologous protein. Finaly, the analysis of immunoblots of homogenates of granulocytes incubated in the presence of the two different proteins was used to show the endocytosis and degradation of heterologous proteins. These results show that isolated EGCs can endocytose and degrade heterologous proteins.
The development and evolution of actin‐containing organelles during spermiogenesis of a primitive nematode
Authors: Noury‐Sraïra Nezha , Gourbault Nicole , Justine Jean‐Lou
Published: August 2004

Summry— Spermatogenesis in the primitive marine nematode Sphaerolaimus hirsutus (Chromadoria, Sphaerolaimidae) was investigated by examining the ultrastructure and cytochemistry. Spermatozoa are lenticular cells of about 15 μm in diameter and are devoid of flagellum and acrosome as in other nematodes. In spermatocytes, dictyosomes produced transient structures, the fibrous body‐membranous organelle complexes (FB‐MO). In spermatids, the FBs were arranged as cartwheel spokes, with the FBs in the centre and the MOs at the periphery. The FBs were first made up of parallel fibres and surrounded by a membrane, then, in a later stage, showed a dense central structure with a surrounding vermiculate region and were devoid of membrane. The FBs contain actin as shown by immunofluorescence using a monoclonal anti‐actin antibody and by affinity cytochemistry using fluorescent phalloidin. MOs contained mainly F‐actin as shown by their labelling by phalloidin. In spermatozoa, the MOs were no longer peripheral but arranged on a ring in the central region of the cell and the FBs disappeared to form the cytoskeleton of the cell outer region. It was assumed, by analogy with the ultrastructure of other nematodes, that this cytoskeleton was made up of major‐sperm‐protein (MSP). Labelling of spermatids of Caenorhabditis elegans also revealed the presence of actin, but cells and actin spots were very can be distinguished. In the few species in which it has been studied (C elegans and Ascaris suum), MSP is thought to constitute in spermatozoa a motile cytoskeleton excluding the presence of actin. However, the present study of Sphaerolaimus shows that the actin cytoskeleton is present during nematode spermiogenesis.
Phosphoinositidase C isozymes in SaOS‐2 cells: Immunocytochemical detection in nuclear and cytoplasmic compartments
Authors: Maraldi Nadir M , Zini Nicoletta , Santi Spartaco , Bavelloni Alberto , Valmori Aurelio , Marmiroli Sandra , Ognibene Andrea
Published: August 2004

Summry— SaOS‐2 cell line presents osteoblastic characteristics which can be modulated by specific agonists involving also phosphoinositide breakdown. In order to determine whether SaOS‐2 cells display a phosphoinositide signalling system not only at the cytosol‐cell membrane level but also, as recently reported for other cell lines, at the nuclear level, a study has been performed to evaluate the phosphoinositidase C (PIC) activity and to localize different isoforms of PIC in nuclear and cytoplasmic compartments. By immunochemicals methods, and by confocal and electron microscope immunocytochemistry, both PIC β1 and γ1 have been detected in the nucleus, while only PIC γ1 was found in the cytoplasm. A specific association with the inner nuclear matrix has been demonstrated for PIC β1 and γ1; this latter resulted, on the other hand, in relationship with cytoskeletal filaments after high salt extraction. These findings suggest that these enzymes are not completely soluble but functionally related with cytoskeletal and nucleoskeletal structures.
Localization and partial characterization of a rat ovarian granulose cell protein with a monoclonal antibody
Authors: Russinova Angelina , Vassilev Alex , Davidoff Michail
Published: August 2004

Summry— Hybridoma cell lines were obtained from mouse splenocytes sensitized to granulosa cells collected from rat ovaries after gonadotropin stimulation. A monoclonal antibody (5G5) was obtained which reacted with granulosa cells and showed a positive reaction with serum‐free conditioned medium containing granulosa cell secreted proteins. Immoblotting of the conditioned medium and light‐ and electron‐microscopic immunocytochemistry of rat ovary show that mAb 5G5 is directed against a 59‐kDa protein which is located on the plasma membrane of granulosa cells. Furthermore, the immunoreactivity of the granulosa cells depends both on the degree of follicle development and on the position of the granulosa cells within the follicles. Strong immunoreactivity was observed in the innermost granulosa cell layers, close to the oocyte and the antral cavity. The results obtained show that mAb 5G5 is a useful marker of a 59‐kDa granulosa cell protein which might be of importance for the follicle and the occyte maturation.