bluuebunny/crossref_metadata_2025 huggingface dataset:
Decrease in motor performance in children with cancer is independent of the cumulative dose of vincristine
Authors: Hartman Annelies , van den Bos Cor , Stijnen Theo , Pieters Rob
Published: February 2006

BACKGROUND

Impaired performance on motor tasks in children treated for acute lymphoblastic leukemia (ALL) after completion of treatment is often attributed to vincristine (VCR). Whether motor problems persist in other children who receive different cumulative doses of VCR is to the authors' knowledge not known. The objective of the current study was to determine the extent of motor problems in children with ALL, Wilms tumor (WT), B‐cell non‐Hodgkin lymphoma (B‐NHL), and malignant mesenchymal tumors (MMT) and whether these motor problems were related to VCR dose.

METHODS

In 128 children ages 4–12 years who completed treatment after at least 1 year, motor performance was measured using the Movement Assessment Battery for Children (m‐ABC).

RESULTS

The m‐ABC scores of the total study group were significantly lower than those of the normal population (P < 0.001). There were no differences in scores noted between children with ALL, WT, B‐NHL, or MMT. There also were no differences noted between those children with ALL who had received pulses of VCR and steroids during maintenance therapy and those who had not. All groups demonstrated large variability in scores. Scores were not found to be significantly different between those children who had received low (< 20 mg/m2), intermediate (20–40 mg/m2), or high (> 40 mg/m2) cumulative doses of VCR. Cumulative doses of corticosteroids and methotrexate did not affect scores, nor did age at diagnosis and time since the completion of therapy.

CONCLUSIONS

Although motor performance was impaired in all patient groups, no correlation was found between motor performance and the cumulative dose of VCR or other drugs, age, and follow‐up time. Future studies have to address several issues, including whether polymorphisms in drug metabolizing genes or drug target genes explain the significant variability noted in the long‐term motor outcome of children with cancer. Cancer 2006. © 2006 American Cancer Society.
Intranasal interleukin‐12 gene therapy enhanced the activity of ifosfamide against osteosarcoma lung metastases
Authors: Duan Xiaoping , Jia Shu‐Fang , Koshkina Nadezhda , Kleinerman Eugenie S.
Published: February 2006

BACKGROUND

Cyclophosphamide (CTX) and ifosfamide (IFX) are alkylating agents used to treat osteosarcoma (OS). It was previously demonstrated that the sensitivity of OS cells to 4‐hydroperoxycyclophosphamide (4‐HC, the active metabolite of CTX) is augmented by interleukin (IL)‐12 in vitro through a mechanism involving the Fas/FasL pathway. The purpose of these studies was to determine whether this synergistic effect is operational in vivo.

METHODS

Mice were injected intravenously with human LM7 osteosarcoma cells. Treatment was initiated with IFX (2.5 mg/kg intraperitoneally) with or without intranasal polyethylenimine (PEI):IL‐12 gene therapy given twice weekly for 6 weeks.

RESULTS

Expression of IL‐12 protein in the lung was demonstrated in all mice receiving intranasal PEI:IL‐12 but not in control mice or those treated with IFX alone. Increased expression of FasL was detected in lungs of all mice receiving IFX. Both IFX and PEI:IL‐12 alone significantly inhibited lung metastasis when compared with control groups (P < 0.05). However, the most significant tumor effect was observed in mice receiving IFX+PEI:IL‐12 (P < 0.01). Immunohistochemical staining for CD31 and basic fibroblast growth factor (bFGF) and the number of proliferating cells as quantified by proliferating cell nuclear antigen (PCNA) staining were also most significantly decreased in mice receiving combination therapy.

CONCLUSIONS

These data indicate that combining IFX and IL‐12 may have therapeutic potential and that this increased efficacy may be mediated through a mechanism involving the Fas/FasL pathway. Cancer 2006. © 2006 American Cancer Society
Visual, auditory, sensory, and motor impairments in long‐term survivors of hematopoietic stem cell transplantation performed in childhood
Authors: Gurney James G. , Ness Kirsten K. , Rosenthal Joseph , Forman Stephen J. , Bhatia Smita , Baker K. Scott
Published: February 2006

BACKGROUND

Because of treatment‐related toxicity, research is increasingly being focused on long‐term sequelae secondary to hematopoietic stem cell transplantation (HSCT) in survivor populations.

METHODS

This study describes the incidence of auditory, sensory, motor, and visual impairments, including cataracts, among 235 individuals who were treated with HSCT during childhood or adolescence. Outcomes were compared with 705 siblings of childhood cancer survivors. Participants completed a survey with questions on posttransplant organ system impairments. Approximately half of survivors were transplanted when younger than 10 years of age. The median length of followup was 11 years.

RESULTS

The cumulative incidence of cataracts was 36% at 15 years post‐HSCT, although cataracts occurred only in those who received total body irradiation as an HSCT conditioning agent or head irradiation before transplant. Persistent pain was reported by 21% of survivors. Loss of hearing in one or both ears, and legal blindness in one or both eyes, each occurred after transplant in 2% of survivors. Occurrences were uncommon, but survivors were 4.3 times (95% confidence interval [CI]: 2.0–9.4) more likely to report coordination problems, 7.7 times (95% CI: 3.2–18.5) more likely to report chewing or swallowing problems, and 3.5 times (3.5; 95% CI: 1.6–7.9) more likely to report muscle weakness than those in the comparison group. Muscle weakness was strongly associated with positive history of chronic graft‐versus‐host disease.

CONCLUSIONS

Increased risks were found for motor impairments, hearing loss, vision loss, and persistent pain among study participants. Cataracts were a frequent adverse effect, suggesting that close monitoring with appropriate intervention for preservation of vision, particularly among those who received total body irradiation, should be a primary goal in survivors of HSCT performed in childhood. Cancer 2006. © 2006 American Cancer Society.
Neural activity related to self‐ versus externally generated painful stimuli reveals distinct differences in the lateral pain system in a parametric fMRI study
Authors: Helmchen Christoph , Mohr Christian , Erdmann Christian , Binkofski F. , Büchel Christian
Published: February 2006

Self‐generated sensory stimulation can be distinguished from externally generated stimulation that is otherwise identical. To determine how the brain differentiates external from self‐generated noxious stimulation and which structures of the lateral pain system use neural signals to predict the sensory consequences of self‐generated painful stimulation, we used functional magnetic resonance imaging to examine healthy human subjects who received thermal‐contact stimuli with noxious and non‐noxious temperatures on the resting right hand in random order. These stimuli were internally (self‐generated) or externally generated. Two additional conditions served as control conditions: to account for stimulus onset uncertainty, acoustic stimuli preceding the same thermal stimuli were used with variable or fixed delays but without any stimulus‐eliciting movements. Whereas graded pain‐related activity in the insula and secondary somatosensory cortex (SII) was independent of how the stimulus was generated, it was attenuated in the primary somatosensory cortex (SI) during self‐generated stimulation. These data agree with recent concepts of the parallel processing of nociceptive signals to the primary and secondary somatosensory cortices. They also suggest that brain areas that encode pain intensity do not distinguish between internally or externally applied noxious stimuli, i.e., this adaptive biological mechanism prevents harm to the individual. The attenuated activation of SI during self‐generated painful stimulation might be a result of the predictability of the sensory consequences of the pain‐related action. Hum Brain Mapp, 2006. © 2006 Wiley‐Liss, Inc.
The role of folate receptor α in cancer development, progression and treatment: Cause, consequence or innocent bystander?
Authors: Kelemen Linda E.
Published: February 2006

Folate receptor α (FRα) is a membrane‐bound protein with high affinity for binding and transporting physiologic levels of folate into cells. Folate is a basic component of cell metabolism and DNA synthesis and repair, and rapidly dividing cancer cells have an increased requirement for folate to maintain DNA synthesis, an observation supported by the widespread use of antifolates in cancer chemotherapy. FRα levels are high in specific malignant tumors of epithelial origin compared to normal cells, and are positively associated with tumor stage and grade, raising questions of its role in tumor etiology and progression. It has been suggested that FRα might confer a growth advantage to the tumor by modulating folate uptake from serum or by generating regulatory signals. Indeed, cell culture studies show that expression of the FRα gene, FOLR1, is regulated by extracellular folate depletion, increased homocysteine accumulation, steroid hormone concentrations, interaction with specific transcription factors and cytosolic proteins, and possibly genetic mutations. Whether FRα in tumors decreases in vivo among individuals who are folate sufficient, or whether the tumor's machinery sustains FRα levels to meet the increased folate demands of the tumor, has not been studied. Consequently, the significance of carrying a FRα‐positive tumor in the era of folic acid fortification and widespread vitamin supplement use in countries such as Canada and the United States is unknown. Epidemiologic and clinical studies using human tumor specimens are lacking and increasingly needed to understand the role of environmental and genetic influences on FOLR1 expression in tumor etiology and progression. This review summarizes the literature on the complex nature of FOLR1 gene regulation and expression, and suggests future research directions. © 2006 Wiley‐Liss, Inc.
Regional DNA hypermethylation and DNA methyltransferase (DNMT) 1 protein overexpression in both renal tumors and corresponding nontumorous renal tissues
Authors: Arai Eri , Kanai Yae , Ushijima Saori , Fujimoto Hiroyuki , Mukai Kiyoshi , Hirohashi Setsuo
Published: February 2006

To evaluate the significance of altered DNA methylation during renal tumorigenesis, tumorous tissues (T) and corresponding nontumorous renal tissues (N) from 94 patients with renal tumors, and normal renal tissues (C) from 16 patients without renal tumors were investigated. DNA methylation status on CpG islands of the p16, human MutL homologue 1 (hMLH1), von‐Hippel Lindau (VHL) and thrombospondin‐1 (THBS‐1) genes and the methylated in tumor (MINT) ‐1, ‐2, ‐12, ‐25 and ‐31 clones and DNA methyltransferase (DNMT) 1 expression were examined by bisulfite modification and immunohistochemistry, respectively. The average number of methylated CpG islands was significantly higher in N than in C, and was even higher in T. The average number of methylated CpG islands in N was significantly correlated with a higher histological grade of corresponding conventional renal cell carcinomas (RCCs). The average number of methylated CpG islands in RCCs was significantly correlated with macroscopic configuration with extranodular or multinodular growth, higher histological grade, infiltrating growth pattern and vascular involvement. The recurrence‐free survival rate of patients with RCCs showing accumulation of DNA methylation was significantly lower than that of patients not showing this feature. The incidence of nuclear immunoreactivity for DNMT1 tended to be higher in proximal tubules from N than in those from C, and was significantly higher in RCCs. From the viewpoint of altered DNA methylation, N is at the precancerous stage, and N showing accumulation of DNA methylation may generate more malignant RCCs. Regional DNA hypermethylation may be associated with renal tumorigenesis from a precancerous condition to malignant progression and become a predictor of patient prognosis. © 2006 Wiley‐Liss, Inc.
Hepatocyte growth factor and the Met system as a mediator of tumor–stromal interactions
Authors: Matsumoto Kunio , Nakamura Toshikazu
Published: February 2006

Crosstalk between carcinoma cells and host stromal cells such as fibroblasts has a great deal of influence on the invasive and metastatic behavior of cancer cells. The oncogenic action of fibroblasts has been demonstrated through genetic alterations that occur specifically in fibroblasts. Hepatocyte growth factor (HGF), a ligand for the Met receptor tyrosine kinase, plays a definitive role, particularly in the progression to invasive and metastatic cancers, predominantly as a stroma‐derived paracrine mediator. Many types of cancer cells secrete molecules that enhance HGF production in fibroblasts, while fibroblast‐derived HGF, in turn, is a potent stimulator of the invasion of cancer cells. Fibroblast‐specific genetic alterations leading to an overexpression of HGF are associated with the development of epithelial neoplasia and invasive carcinoma. Strategies for targeting the HGF–Met axis are being pursued, in attempts to block the malignant behavior of cancers. In normal tissues, the HGF–Met axis plays diverse roles in organogenesis and in wound healing. The simile that “cancer is a never‐healing wound” appears to be pertinent here. © 2006 Wiley‐Liss, Inc.
Development of a reinforced porcine elastin composite vascular scaffold
Authors: Hinds Monica T. , Rowe Rebecca C. , Ren Zhen , Teach Jeffrey , Wu Ping‐Cheng , Kirkpatrick Sean J. , Breneman Kathryn D. , Gregory Kenton W. , Courtman David W.
Published: February 2006

Elastin, a principal structural component of native arteries, has distinct biological and mechanical advantages when used as a biomaterial; however, its low ultimate tensile strength has limited its use as an arterial conduit. We have developed a scaffold, consisting of a purified elastin tubular conduit strengthened with fibrin bonded layers of acellular small intestinal submucosa (aSIS) for potential use as a small diameter vascular graft. The addition of aSIS increased the ultimate tensile strength of the elastin conduits nine‐fold. Burst pressures for the elastin composite vascular scaffold (1396 ± 309 mmHg) were significantly higher than pure elastin conduits (162 ± 36 mmHg) and comparable to native saphenous veins. The average suture pullout strength of the elastin composite vascular scaffolds was 14.612 ± 3.677 N, significantly higher than the pure elastin conduit (0.402 ± 0.098 N), but comparable to native porcine carotid arteries (13.994 ± 4.344 N). Cyclic circumferential strain testing indicated that the composite scaffolds were capable of withstanding physiological loading conditions for at least 83 h. Implantation of the elastin composites as carotid interposition grafts in swine demonstrated its superiority to clinically acceptable ePTFE with significantly longer average patency times of 5.23 h compared to 4.15 h. We have developed a biologically based elastin scaffold with suitable mechanical properties and low thrombogenicity for in vivo implantation, and with the potential for cellular repopulation and host integration reestablishing an appropriate elastic artery. © 2006 Wiley Periodicals, Inc. J Biomed Mater Res, 2006
Substrate specificity of soluble and membrane‐associated ADP‐ribosyltransferase ART2.1
Authors: Zheng Xuexiu , Morrison Alan R. , Chung An‐Sik , Moss Joel , Bortell Rita
Published: February 2006

ADP‐ribosyltransferases (ARTs) are a family of enzymes that catalyze the covalent transfer of an ADP‐ribose moiety, derived from NAD, to an amino acid of an acceptor protein, thereby altering its function. To date, little information is available on the protein target specificity of different ART family members. ART2 is a T‐cell‐specific transferase, attached to the cell surface by a glycosylphosphatidylinositol (GPI) anchor, and also found in serum. Here we investigated the role of ART2 localization in serum or on the cell surface, or solubilized with detergents or enzymes, on its target protein specificity. We found that detergent solubilization of cell membranes, or release of ART2 by phosphoinositide‐specific phospholipase C treatment, altered the ability of ART2 to ADP‐ribosylate high or low molecular weight histone proteins. Similarly, soluble recombinant ART2 (lacking the GPI anchor) showed a different histone specificity than did cell‐bound ART2. When soluble ART2 was incubated with serum proteins in the presence of [32P]‐labeled NAD, several serum proteins were ADP‐ribosylated in a thiol‐specific manner. Mass spectrometry of labeled proteins identified albumin and transferrin as ADP‐ribosylated proteins in serum. Collectively, these studies reveal that the membrane or solution environment of ART2 plays a pivotal role in determining its substrate specificity. J. Cell. Biochem. 98: 851–860, 2006. © 2006 Wiley‐Liss, Inc.
Role of AP1 element in the activation of human eNOS promoter by lysophosphatidycholine
Authors: Xing Feiyue , Jiang Yong , Liu Jing , Zhao Kesen , Mo Yongyan , Qin Qinghe , Wang Jingzhen , Ouyang Jianming , Zeng Yaoying
Published: February 2006

Human endothelial nitric oxide synthase (eNOS) plays a crucial role in maintaining blood pressure homeostasis and vascular integrity. It, therefore, is very essential to elucidate the regulation of it. In the current study, a red fluorescent protein (RFP) reporter system containing human eNOS promoter was first constructed, being characteristics of real time morphologic and quantitative analysis for the same sample. It was observed by DNA sequence deletion that 68% of the basal activity of the promoter was controlled by the region from −1 to −166 bp, and 32% of it was dependent on the region from −1,033 to −1,600 bp. The mutation of SSRE element (−999∼−994 bp) and wild‐type SSRE decoy oligodeoxynucleotides (ODN) did not alter the basal activity and the stimulating activity by lysophosphatidycholine (LPC). The mutation of upstream AP1 element (−1,530∼−1,524 bp) did not affect the basal activity, but resulted in near 30% reduction in the stimulating activity by LPC. Moreover, wild‐type AP1 decoy ODN also remarkably attenuated it. It was proved by EMSA analysis that LPC indeed enhanced the activity of AP1 transcriptional factor binding to AP1 element. However, the role of AP1 was dependent on the presence of SP1, which was proved by the combining mutation of AP1 with SP1. The mutation of downstream AP1 element (−662∼−656 bp) had no influence on the basal and stimulating activities by LPC. These results strongly suggest that the main functional region of the promoter is from −1 bp to −166 bp, that the upstream AP1 participates in the activation of the promoter by LPC on the premise of the presence of SP1, and that the downstream AP1 and SSRE do not involve the basal and stimulating activity by LPC. J. Cell. Biochem. 98: 872–884, 2006. © 2006 Wiley‐Liss, Inc.
Expression of DNMT3A transcripts and nucleolar localization of DNMT3A protein in human testicular and fibroblast cells suggest a role for de novo DNA methylation in nucleolar inactivation
Authors: Galetzka Danuta , Tralau Tim , Stein Raimund , Haaf Thomas
Published: February 2006

Transcriptional silencing during differentiation of human male germ cells and serum starvation of human fibroblasts is controlled by epigenetic mechanisms that involve de novo DNA methylation. It is associated with high expression of different trancripts of the DNA methyltransferase 3A (DNMT3A) gene that encode two isoforms with de novo methyltransferase activity and one without catalytic activity. Western blots revealed that DNMT3A protein (with catalytic domain) is present at low levels in several tissues and at increased levels in testicular cells and growth‐arrested fibroblasts. Immunofluorescence experiments localized DNMT3A to discrete nucleolar foci in B spermatogonia and resting fibroblasts. The data here suggest a role for de novo DNA methylation in nucleolar inactivation. J. Cell. Biochem. 98: 885–894, 2006. © 2006 Wiley‐Liss, Inc.
Functional and structural properties of stannin: Roles in cellular growth, selective toxicity, and mitochondrial responses to injury
Authors: Billingsley M.L. , Yun J. , Reese B.E. , Davidson C.E. , Buck‐Koehntop B.A. , Veglia G.
Published: February 2006

Stannin (Snn) was discovered using subtractive hybridization methodology designed to find gene products related to selective organotin toxicity and apoptosis. The cDNAs for Snn were first isolated from brain tissues sensitive to trimethyltin, and were subsequently used to localize, characterize, and identify genomic DNA, and other gene products of Snn. Snn is a highly conserved, 88 amino acid protein found primarily in vertebrates. There is a minor divergence in the C‐terminal sequence between amphibians and primates, but a nearly complete conservation of the first 60 residues in all vertebrates sequenced to date. Snn is a membrane‐bound protein and is localized, in part, to the mitochondria and other vesicular organelles, suggesting that both localization and conservation are significant for the overall function of the protein. The structure of Snn in a micellar environment and its architecture in lipid bilayers have been determined using a combination of solution and solid‐state NMR, respectively. Snn structure comprised a single transmembrane domain (residues 10–33), a 28‐residue linker region from residues 34–60 that contains a conserved CXC metal binding motif and a putative 14‐3‐3ξ binding region, and a cytoplasmic helix (residues 61–79), which is partially embedded into the membrane. Of primary interest is understanding how this highly‐conserved peptide with an interesting structure and cellular localization transmits both normal and potentially toxic signals within the cell. Evidence to date suggests that organotins such as trimethyltin interact with the CXC region of Snn, which is vicinal to the putative 14‐3‐3 binding site. In vitro transfection analyses and microarray experiments have inferred a possible role of Snn in several key signaling systems, including activation of the p38‐ERK cascade, p53‐dependent pathways, and 14‐3‐3ξ protein‐mediated processes. TNFα can induce Snn mRNA expression in endothelial cells in a PKC‐ε dependent manner. Studies with Snn siRNA suggest that this protein may be involved in growth regulation, since inhibition of Snn expression alone leads to reduced endothelial cells growth and induction of COP‐1, a negative regulator of p53 function. A key piece of the puzzle, however, is how and why such a highly‐conserved protein, localized to mitochondria, interacts with other regulatory proteins to alter growth and apoptosis. By knowing the structure, location, and possible signaling pathways involved, we propose that Snn constitutes an important sensor of mitochondrial damage, and plays a key role in the mediation of cross‐talk between mitochondrial and nuclear compartments in specific cell types. J. Cell. Biochem. 98: 243–250, 2006. © 2006 Wiley‐Liss, Inc.
Recombinant human uteroglobin/CC10 inhibits the adhesion and migration of primary human endothelial cells via specific and saturable binding to fibronectin
Authors: Antico Giovanni , Lingen Mark W. , Sassano Antonella , Melby James , Welch Richard W. , Fiore Stefano , Pilon Aprile L. , Miele Lucio
Published: February 2006

Uteroglobin (UG) or Clara Cell 10 kDa protein (CC10) is a small, stable, epithelial secretory anti‐inflammatory protein. Uteroglobin has been shown to inhibit neointimal formation in vivo after balloon angioplasty through an unknown mechanism. An interaction between UG and plasma fibronectin (Fn) has been demonstrated in mice. Since Fn plays a key role in endothelial cell (EC) migration and angiogenesis, we investigated whether recombinant human UG (rhUG) affects EC migration via Fn binding. In this report, we show a saturable binding of rhUG to Fn depending on Fn conformation and that rhUG is covalently cross‐linked to Fn by transglutaminase (TGase). Additionally, our study highlights that rhUG can also bind to exogenously added or self‐secreted Fn on the membrane of human primary microvascular endothelial cells (HMVEC), although these complexes are weakly associated with the plasmalemma. Upon the interaction with Fn in solid phase, rhUG strongly inhibits HMVEC attachment on Fn, but not on other ECM proteins. Consequently, rhUG also inhibits cell migration in a dose dependent fashion (I.C.50 = 65 nM) and hinders the “wound healing” in vitro. The small size, stability and human tolerability of rhUG suggest that rhUG in slow‐release form or genetically delivered could be used in humans to modulate cell/Fn interactions in the context of tumor microenvironment or in the context of inflammation and fibrosis. J. Cell. Physiol. 207: 553–561, 2006. © 2006 Wiley‐Liss, Inc.
Interleukin‐1 beta induction of matrix metalloproteinase‐1 transcription in chondrocytes requires ERK‐dependent activation of CCAAT enhancer‐binding protein‐beta
Authors: Raymond Lauren , Eck Sarah , Mollmark Jessica , Hays Ezra , Tomek Ivan , Kantor Stephen , Elliott Sarah , Vincenti Matthew
Published: February 2006

Interleukin‐1 beta (IL‐1β) is a central mediator of inflammation and connective tissue destruction in rheumatoid arthritis. IL‐1β activates articular chondrocytes to produce matrix metalloproteinase‐1 (MMP‐1), an enzyme capable of dismantling the collagen scaffold of articular cartilage. To define the transcription factors and signaling intermediates that activate MMP‐1 transcription in chondrocytes, we performed transient transfection of MMP‐1 promoter constructs followed by reporter assays. These studies identified an IL‐1β‐responsive region of the human MMP‐1 promoter that contains a consensus CCAAT enhancer‐binding protein (C/EBP) binding site. Deletion of this site reduced overall transcriptional activity of the MMP‐1 promoter, as well as decreased fold induction by IL‐1β. IL‐1β stimulation of chondrocytes increased binding of C/EBP‐β to the MMP‐1 C/EBP site. Extracellular signal regulated kinase (ERK) pathway‐dependent phosphorylation of C/EBP‐β on threonine 235 activates this transcription factor. Here we show that IL‐1β stimulation of chondrocytes induced phosphorylation of C/EBP‐β on threonine 235, and that the ERK pathway inhibitor PD98059 reduced this phosphorylation. We further show that PD98059 reduces IL‐1β‐induced MMP‐1 mRNA expression in chondrocytes. Moreover, inhibition of the ERK pathway by expression of dominant‐negative forms of ERK1 and ERK2 impaired the ability of IL‐1β to transactivate the MMP‐1 promoter. Our findings demonstrate a novel role for C/EBP‐β in IL‐1β‐induced connective tissue disease and define a new nuclear target for the ERK pathway in MMP‐1 gene activation. J. Cell. Physiol. © 2006 Wiley‐Liss, Inc.
Src oncogene activates MMP‐2 expression via the ERK/Sp1 pathway
Authors: Kuo LunYu , Chang Hui‐Chiu , Leu Tzeng‐Horng , Maa Ming‐Chie , Hung Wen‐Chun
Published: February 2006

Previous studies demonstrated that activation of Src oncogene increased matrix metalloproteinase‐2 (MMP‐2) expression in various types of cells. In this study, we elucidated the underlying mechanism of Src‐induced MMP‐2. We first used murine fibroblast cell line C3H10T1/2 and its v‐Src transfectant IV5 to address this issue. RT‐PCR and promoter activity assay indicated that Src stimulated MMP‐2 via transcriptional activation. Transfection of constitutively active Src into C3H10T1/2 cells also stimulated MMP‐2 mRNA expression. Deletion and mutation analysis indicated that the Sp1 site located within the −91/−84 region of human MMP‐2 promoter is the major responsive element for Src. Electrophoresis mobility shift assays showed that Src enhanced the binding of Sp1 to this consensus site to stimulate MMP‐2 gene expression. We next investigated the signaling pathway that mediated the effect of Src on MMP‐2. Our data showed that extracellular signal‐regulated kinase (ERK) pathway inhibitor PD98059, but not the c‐Jun N‐terminal kinase (JNK) inhibitor SP600125, p38 kinase inhibitor SB203580, and PI3K inhibitor wortmannin, attenuated Src‐induced MMP‐2 promoter activity. These inhibitors did not show significant effect on basal MMP‐2 promoter activity in C3H10T1/2 cells. In addition, the dominant negative mutant of ERK‐2 suppressed the activation of MMP‐2 by Src. Treatment of PD98059 or an Src specific inhibitor PP1 reduced Sp1 DNA binding activity in IV5 cells. Taken together, our results strongly suggest that Src induces MMP‐2 expression via transcription activation and the ERK/Sp1 signaling pathway is involved in this process. J. Cell. Physiol. © 2006 Wiley‐Liss, Inc.
Off‐line combination of reversed‐phase liquid chromatography and laser desorption/ionization time‐of‐flight mass spectrometry with seamless post‐source decay fragment ion analysis for characterization of square‐planar nickel(II) complexes
Authors: Řehulka Pavel , Popkov Alexander , Nádvorník Milan , Planeta Josef , Mazanec Karel , Chmelík Josef
Published: February 2006

Characterization of square‐planar nickel(II) complexes of the Schiff base of (S)‐N‐benzylproline (2‐benzoylphenyl)amide and various amino acids that are used as efficient α‐amino acids synthons was carried out using laser desorption/ionization time‐of‐flight mass spectrometry (LDI‐TOF MS) in off‐line combination with liquid chromatography. A mixture of four square‐planar nickel(II) complexes was separated using reversed‐phase liquid chromatography (RPLC) and the separated fractions from the chromatographic run were spotted on the metal target directly from the column outlet using a lab‐made sample deposition device. The separated fractions were then analyzed by LDI‐TOF MS. Seamless postsource decay (sPSD) fragment ion analysis was used for their structural characterization, which made possible the confirmation of expected chemical structures of the analyzed compounds. The off‐line combination of the separation by RPLC and analysis by LDI‐TOF MS allowed successful separation, sensitive detection and structure elucidation of the square‐planar nickel(II) complexes. Copyright © 2006 John Wiley & Sons, Ltd.
Analysis of fungal cyclopentenone derivatives from Hygrophorus spp. by liquid chromatography/electrospray‐tandem mass spectrometry
Authors: Lübken Tilo , Arnold Norbert , Wessjohann Ludger , Böttcher Christoph , Schmidt Jürgen
Published: February 2006

Fruitbodies of the genus Hygrophorus (Basidiomycetes) contain a series of anti‐biologically active compounds. These substances named hygrophorones possess a cyclopentenone skeleton. LC/ESI‐MS/MS presents a valuable tool for the identification of such compounds. The mass spectral behaviour of typical selected members of this group under positive and negative ion electrospray conditions is discussed. Using the ESI collision‐induced dissociation (CID) mass spectra of the [M + H]+ and [M − H]− ions, respectively, the compounds can be classified with respect to the substitution pattern at the cyclopentenone ring and the type of oxygenation at C‐6 (hydroxy/acetoxy or oxo function) of the side chain. The elemental composition of the fragment ions was determined by ESI‐QqTOF measurements. Thus, in case of the negative ion CID mass spectra an unusual loss of CO2 from the deprotonated molecular ions could be observed. Copyright © 2006 John Wiley & Sons, Ltd.
Canine ACL fibroblast integrin expression and cell alignment in response to cyclic tensile strain in three‐dimensional collagen gels
Authors: Henshaw D. Ross , Attia Erik , Bhargava Madhu , Hannafin Jo A.
Published: February 2006

Tissue‐engineered ligament substitutes have the potential to become an alternative graft source for ligament reconstruction. If this approach is to become viable, one must first understand and define the mechanisms responsible for creation, maintenance, and remodeling of the native anterior cruciate ligament. It is well accepted that mechanical load alters fibroblast phenotypic expression in a variety of cell sources; however, the mechanosensitive pathways responsible for alteration in matrix production, remodeling, and alignment are unknown. We hypothesize that cell surface integrins play a role in this mechanotransduction process, and as such respond to application of cyclic tensile load. Linear 3D collagen gels containing canine ACL fibroblasts were created in Flexercell Tissue‐Train Culture Plates. Gels were untethered (control), tethered without external strain (tethered), or tethered and exposed to 2.5% cyclic strain for 2 h per day for 4 days (strain). Quantitation of α1, α5, and β1 integrin subunit was performed using flow cytometry. Cell and matrix alignment was studied using light, polarized light, and fluorescent microscopy. Expression of α5 and β1 integrin subunits was increased significantly in fibroblasts in tethered and strained 3D collagen gels compared with the control, unloaded constructs (p < 0.05). These integrins are known to function as mechanotransducers in other tissues, implicating a similar role in mechanotransduction in ACL fibroblasts. Histologic analysis of the tethered and strained gels demonstrated a linear arrangement of cells and parallel collagen fibril architecture. In contrast, cell distribution and collagen alignment were disorganized in the control, unloaded gels. The alignment of cells and collagen in the 3D gels parallel to applied strain is similar to the in vivo state. These data add to our understanding of the behavior of ACL fibroblasts in vitro. The ability to manipulate signal transduction pathways may enhance our ability to engineer implantable ACL grafts or to modify ACL healing response. © 2006 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 24:481–490, 2006
The effect of thrombin on ACL fibroblast interactions with collagen hydrogels
Authors: Murray M.M. , Forsythe B. , Chen F. , Lee S.J. , Yoo J.J. , Atala A. , Steinert A.
Published: February 2006

Premature loss of provisional scaffold formation has been identified as one of the factors responsible for poor healing of intraarticular tissues. To address this deficiency, substitute provisional scaffolds are being developed. The function of these scaffolds can be enhanced by the addition of specific extracellular matrix proteins. In this study, it was hypothesized that the addition of thrombin to a provisional scaffold material would result in increases in cell proliferation, collagen production, and cell migration within the scaffold. These three parameters are thought to be critical components of wound healing. Gels containing fibrin and collagen supplemented with either 0, 10.5, 21, or 42 U/mL of thrombin were placed in contact with explants of tissue from the anterior cruciate ligament. The addition of thrombin stimulated cell migration at low concentrations and impaired migration at higher concentrations, and had no significant effect on cell proliferation or collagen production. The use of all concentrations of thrombin resulted in mechanically weaker gels. Thus, the use of thrombin to optimize a collagen‐platelet rich plasma (PRP) provisional scaffold must be done with caution, and use of high concentrations of thrombin (>42 IU/mL) should be avoided specifically in situations where gel strength or cell ingrowth is important. Use of low concentrations of thrombin (10.5 IU/mL) may be beneficial in applications where a faster set time and enhanced cell migration are desirable and the gel mechanical strength is of secondary importance. © 2006 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 24:508–515, 2006
When service fails: The role of the salesperson and the customer
Authors: Chang Chia‐Chi
Published: February 2006

Service failures and consequent recoveries have been identified as a critical determinant of customer retention. Hence, effective service recovery programs warrant further exploration. In particular, with the current emphasis on relationship selling, salespeople should learn how to improvise to serve their customers effectively, including solving problems when service failure occurs. Therefore, alternatives to current common practices in industry should be investigated. The results of the present study suggest that by providing customers a choice of service recovery options, salespeople can augment customers' sense of control and ultimately their satisfaction with the service recovery and the overall service transaction. The investigation also proposed that customer satisfaction would be contingent upon the importance of the service. However, study findings revealed that this was not the case. Implications of the findings are also discussed. © 2006 Wiley Periodicals, Inc.
Overcoming the illusion of will and self‐fabrication: Going beyond naïve subjective personal introspection to an unconscious/conscious theory of behavior explanation
Authors: Woodside Arch G.
Published: February 2006

Naïve subjective personal introspection includes the failure to recognize the confirmability of one's own attitudes and personal meanings learned explicitly from self‐examining such topics and explaining one's own behavior. Unconscious/conscious theory of behavior explanation follows from unifying the research on unintended thought–behavior with folk explanations of behavior. This article describes advances in research confirming own attitudes and personal meaning and suggests the need for applying multiple methods to overcome the fundamental attribution error, inherent cultural prejudices, and the general bias toward self‐fabrication. The discussion is valuable for achieving a deep understanding of how customers think, advancing from subjective to confirmatory personal introspection, and understanding the need to apply research tools useful for enlightening knowledge and overcoming the inherent bias within subjective personal introspection. © 2006 Wiley Periodicals, Inc.
Blood donations, iron stores, and risk of Parkinson's disease
Authors: Logroscino Giancarlo , Chen Honglei , Wing Al , Ascherio Alberto
Published: February 2006

Iron overload and systemic iron stores may be important in the pathogenesis of Parkinson's disease (PD). We therefore examined the association between blood donations, which reduce body iron stores, and risk of PD in the Health Professionals Follow‐Up Study, a large cohort investigation of U.S. men. Our hypothesis was that blood donation reduces the risk of PD by lowering systemic iron stores. Although the number of blood donations was inversely related to the ferritin levels in a subsample of the study population, no association was found between the number of blood donations and risk of PD (P for trend = 0.6). Unexpectedly, the risk of PD was higher among men who reported recent multiple blood donations (P for trend = 0.05). The results of this study do not support the hypothesis that reduced systemic iron stores lower the risk of PD. © 2006 Movement Disorder Society
Automatic selection of arterial input function using cluster analysis
Authors: Mouridsen Kim , Christensen Søren , Gyldensted Louise , Østergaard Leif
Published: February 2006

Quantification of cerebral blood flow (CBF) using dynamic susceptibility contrast MRI requires determination of the arterial input function (AIF) representing the delivery of intravascular tracer to tissue. This is typically accomplished manually by inspection of concentration time curves (CTCs) in regions containing the ICA, VA, and MCA. This is, however, a time consuming and operator dependent procedure. We suggest a completely automatic procedure for establishing the AIF based on a cluster analysis algorithm. In 20 normal subjects CBF maps calculated in 2 slices by the automatic procedure were compared to maps obtained with AIFs selected individually by 7 experienced operators. The average manual to automatic CBF ratio was 1.03 ± 0.15 in the lower slice and 1.05 ± 0.12 in the upper slice, demonstrating excellent agreement between the manual and automatic method. The algorithm provides means for objectively assessing AIF candidates in local AIF search algorithms designed to reduce bias due to delay and dispersion. Given the reproducibility and speed (10 s) of the automatic method, we speculate that it will greatly improve the accuracy of perfusion images and facilitate their use in clinical diagnosis and decision‐making, particularly in acute stroke but also in cerebrovascular disease in general. Magn Reson Med, 2006. © 2006 Wiley‐Liss, Inc.
Mice with a deletion in the first intron of the Col1a1 gene develop dissection and rupture of aorta in the absence of aneurysms: High‐resolution magnetic resonance imaging, at 4.7 T, of the aorta and cerebral arteries
Authors: Marjamaa J. , Tulamo R. , Abo‐Ramadan U. , Hakovirta H. , Frösen J. , Rahkonen O. , Niemelä M. , Bornstein P. , Penttinen R. , Kangasniemi M.
Published: February 2006

Deletion of the majority of the first intron of the Col1a1 gene in mice leads to decreased type I collagen synthesis and content in the aortic wall. In 54% of cases, mice homozygous for the Col1a1 mutation die of thoracic hemorrhage by the age of 18 months. It is unknown whether the fatal bleeding results from an acute dissection of the aortic wall or a gradually developing dilatation of the medial layer prior to rupture. We optimized high‐resolution MRI methods using a 4.7 T MR scanner to obtain in vivo images of the entire mouse aorta. The MR images were acquired in three imaging planes using gradient echo, spin echo, and spin echo with inversion recovery pulse sequences with a maximum in‐plane resolution of 68 × 68 μm and acquisition times less than 10 min. In five Col1a1 mutated mice aged 16 months, the MR images showed no signs of aneurysmal dilatation, wall defects, or former dissection, suggesting that the mechanism for aortic rupture is an acute dissection of the aortic medial layer. Cerebral arteries were imaged using a three‐dimensional time of fight pulse sequence. The resolution of 73 × 73 × 94 μm showed normal cerebral arteries. Histology showed a 22% thinner cerebral artery wall in Col1a1 mutated mice. Magn Reson Med, 2006. © 2006 Wiley‐Liss, Inc.
In vivo detection of brain glycine with echo‐time‐averaged 1H magnetic resonance spectroscopy at 4.0 T
Authors: Prescot Andrew P. , Frederick Blaise deB , Wang Liqun , Brown John , Jensen J. Eric , Kaufman Marc J. , Renshaw Perry F.
Published: February 2006

A single‐voxel proton magnetic resonance spectroscopy (1H‐MRS) method is described that enables the in vivo measurement of endogenous brain glycine (Gly) levels in human subjects. At 4.0 T, TE‐averaging 1H‐MRS dramatically attenuates the overlapping myo‐inositol (mI) resonances at 3.52 ppm, permitting a more reliable measure of the Gly singlet peak. This methodology initially is described and tested in phantoms. The phantom data infers that the 3.55‐ppm peak predominantly is Gly with a smaller contribution from mI. The composite resonance thus is differentiated from pure Gly and mI and is labeled Gly*. The mI contribution was calculated as <2% of the total Gly* signal for a 1:1 mI/Gly mixture. The technique subsequently was used to acquire TE‐averaged 1H‐MRS data from the occipital cortex of healthy control subjects. The resultant spectra closely resembled experimental phantom data. LC‐model analysis provided a means for quantifying TE‐averaged 1H‐MRS spectra and a mean test–retest variability measure of 15% was established for brain Gly* levels in studies of six healthy subjects. Magn Reson Med, 2006. © 2006 Wiley‐Liss, Inc.
Analysis of residual coronary artery motion for breath hold and navigator approaches using real‐time coronary MRI
Authors: Fischer R. W. , Botnar R. M. , Nehrke K. , Boesiger P. , Manning W. J. , Peters D. C.
Published: February 2006

Coronary artery MRI methods utilize breath holds, or diaphragmatic navigators, to compensate for respiratory motion. To increase image quality and navigator (NAV) gating efficiency, slice tracking is used, with more sophisticated affine motion models recently introduced. This study assesses the extent of remaining coronary artery motion in free breathing NAV and single and multi breath hold coronary artery MRI. Additionally, the effect of the NAV gating window size was examined. To visualize and measure the respiratory induced motion, an image containing a coronary artery cross section was acquired at each heartbeat. The amount of residual coronary artery displacement was used as a direct measure for the performance of the respiratory motion correction method. Free breathing studies with motion compensation (slice tracking with 5 mm gating window) had a similar amount of residual motion (0.76 ± 0.17 mm) as a single breath hold (0.52 ± 0.20 mm) and were superior to multiple breath holds (1.22 ± 0.60 mm). Affine NAV methods allowed for larger gating windows (∼10 mm windows) with similar residual motion (0.74 ± 0.17 mm). In this healthy adult cohort (N = 10), free‐breathing NAV methods offered respiratory motion suppression similar to a single breath hold. Magn Reson Med, 2006. © 2006 Wiley‐Liss, Inc.
Auto‐calibrated parallel spiral imaging
Authors: Heberlein and Keith , Hu Xiaoping
Published: February 2006

This work describes an auto‐calibrated method for parallel imaging with spiral trajectory. The method is a k‐space approach where an interpolation kernel, accounting for coil sensitivity factors, is derived from experimental data and used to interpolate the reduced data set in parallel imaging to estimate the missing k‐space data. For the case of spiral imaging, this interpolation kernel is defined along radial directions so that missing spiral interleaves can be estimated directly from neighboring interleaves. This kernel is invariant along the radial direction but varies azimuthally. Therefore, the k‐space is divided into angular sectors and sector‐specific kernels are used. It is demonstrated experimentally that relatively few sectors are sufficient for accurate reconstruction, allowing for efficient implementation. The interpolation kernels can be derived either from a separate calibration scan or self‐calibration data available with a dual‐density spiral acquisition. The reconstruction method is implemented with two sampling strategies and experimentally demonstrated to be robust. Magn Reson Med, 2006. © 2006 Wiley‐Liss, Inc.
Peripheral neuropathy in an outpatient cohort of patients with Sjögren's syndrome
Authors: Lopate Glenn , Pestronk Alan , Al‐Lozi Muhammad , Lynch Timothy , Florence Julaine , Miller Timothy , Levine Todd , Rampy Tom , Beson Brent , Ramneantu Irena
Published: February 2006

Peripheral neuropathy is common in patients with Sjögren's syndrome (SS), but its precise prevalence is unknown. Most prior studies were conducted at neurology or rheumatology specialty clinics and likely selected for a more severely affected population. We evaluated 22 SS patients and 10 controls for evidence of neuropathy in an outpatient setting at a regional meeting of the Sjögren's Syndrome Foundation. We performed neurological examinations and nerve conduction studies (NCSs) and measured serum antinuclear antibody (ANA) and SS‐A and SS‐B antibody levels. Participants filled out a questionnaire pertaining to symptoms, diagnosis, and treatment. We found that signs and symptoms related to small axons were more common in patients with SS than in controls. Complaints of painful distal paresthesias in the feet were noted in 59% of patients but in only 10% of controls, and of abnormal sweating in 41% and 0%, respectively. Examination revealed decreased pinprick sensation in 64% of patients with SS, but in only 30% of controls. Overall, 45% of the patients but none of the controls were thought to have an isolated small‐fiber neuropathy. Large‐fiber dysfunction (as measured by testing vibration, deep tendon reflexes, and NCSs) was similar between the two groups. We conclude that small‐fiber neuropathy is common in patients with SS. Muscle Nerve 2006