bluuebunny/crossref_metadata_2025 huggingface dataset:
The interplay between parental beliefs about children's emotions and parental stress impacts children's attachment security
Authors: Stelter Rebecca L. , Halberstadt Amy G.
Published: May 2010

This study investigated how parental beliefs about children's emotions and parental stress relate to children's feelings of security in the parent–child relationship. Models predicting direct effects of parental beliefs and parental stress, and moderating effects of parental stress on the relationship between parental beliefs and children's feelings of security were tested. Participants were 85 African American, European American, and Lumbee American Indian 4th and 5th grade children and one of their parents. Children reported their feelings of security in the parent–child relationship; parents independently reported on their beliefs and their stress. Parental stress moderated relationships between three of the four parental beliefs about the value of children's emotions and children's attachment security. When parent stress was low, parental beliefs accepting and valuing children's emotions were not related to children's feelings of security; when parent stress was high, however, parental beliefs accepting and valuing children's emotions were related to children's feelings of security. These findings highlight the importance of examining parental beliefs and stress together for children's attachment security. Copyright © 2010 John Wiley & Sons, Ltd.
VEGF receptor binding peptide‐linked high mobility box group‐1 box A as a targeting gene carrier for hypoxic endothelial cells
Authors: Han Jee Seung , Kim Hyun Ah , Lee Sanghyun , Lee Minhyung
Published: May 2010

High mobility group box‐1 (HMGB‐1) is a nuclear protein that can bind to and condense plasmid DNA. In this study, we developed a recombinant VEGF receptor binding peptide (VRBP) linked to HMGB‐1 box A (VRBP‐HMGB1A) as a targeting gene carrier to hypoxic endothelial cells. Hypoxic endothelial cells in ischemic tissues of solid tumors are important targets for gene therapy. A recombinant VRBP‐HMGB1A expression vector, pET21a‐VRBP‐HMGB1A was constructed. VRBP‐HMGB1A was over‐expressed in BL21 strain and purified by nickel‐chelate affinity chromatography. Complex formation between VRBP‐HMGB1A and pCMV‐Luc was confirmed by gel retardation assay. pCMV‐Luc was retarded completely at a 2/1 weight ratio (peptide/plasmid). For transfection assays, calf pulmonary artery endothelial (CPAE) cells were incubated under hypoxia for 24 h, prior to transfection to induce the VEGF receptors on the cells. VRBP‐HMGB1A/pCMV‐Luc complexes were transfected to hypoxic CPAE cells. The highest transfection efficiency was at a 30/1 weight ratio (peptide/plasmid). In addition, VRBP‐HMGB1A had higher efficiency than poly‐L‐lysine (PLL) specifically in hypoxic CPAE cells, However, VRBP‐HMGB1A had lower efficiency than PLL in 293, H9C2, and normoxic CPAE cells. In MTT assay, VRBP‐HMGB1A was less toxic than PLL to cells. In conclusion, VRBP‐HMGB1A is a potential gene carrier for targeting hypoxic endothelial cells and thus, may be useful for cancer gene therapy. J. Cell. Biochem. 110: 1094–1100, 2010. Published 2010 Wiley‐Liss, Inc.
Transcriptional suppression of mir‐29b‐1/mir‐29a promoter by c‐Myc, hedgehog, and NF‐kappaB
Authors: Mott Justin L. , Kurita Satoshi , Cazanave Sophie C. , Bronk Steven F. , Werneburg Nathan W. , Fernandez‐Zapico Martin E.
Published: May 2010

MicroRNAs regulate pathways contributing to oncogenesis, and thus the mechanisms causing dysregulation of microRNA expression in cancer are of significant interest. Mature mir‐29b levels are decreased in malignant cells, and this alteration promotes the malignant phenotype, including apoptosis resistance. However, the mechanism responsible for mir‐29b suppression is unknown. Here, we examined mir‐29 expression from chromosome 7q32 using cholangiocarcinoma cells as a model for mir‐29b downregulation. Using 5′ rapid amplification of cDNA ends, the transcriptional start site was identified for this microRNA locus. Computational analysis revealed the presence of two putative E‐box (Myc‐binding) sites, a Gli‐binding site, and four NF‐κB‐binding sites in the region flanking the transcriptional start site. Promoter activity in cholangiocarcinoma cells was repressed by transfection with c‐Myc, consistent with reports in other cell types. Treatment with the hedgehog inhibitor cyclopamine, which blocks smoothened signaling, increased the activity of the promoter and expression of mature mir‐29b. Mutagenesis analysis and gel shift data are consistent with a direct binding of Gli to the mir‐29 promoter. Finally, activation of NF‐κB signaling, via ligation of Toll‐like receptors, also repressed mir‐29b expression and promoter function. Of note, activation of hedgehog, Toll‐like receptor, and c‐Myc signaling protected cholangiocytes from TRAIL‐induced apoptosis. Thus, in addition to c‐Myc, mir‐29 expression can be suppressed by hedgehog signaling and inflammatory pathways, both commonly activated in the genesis of human malignancies. J. Cell. Biochem. 110: 1155–1164, 2010. Published 2010 Wiley‐Liss, Inc.
ERp57 is up‐regulated in free fatty acids‐induced steatotic L‐02 cells and human nonalcoholic fatty livers
Authors: Wang Hui , Chan Ping‐Kei , Pan Si‐Yuan , Kwon Kwok‐Ho , Ye Yan , Chu Jian‐Hong , Fong Wang‐Fun , Tsui Wilson M.S. , Yu Zhi‐Ling
Published: May 2010

Pathogenesis of nonalcoholic fatty liver disease (NAFLD) is not clear. In this study we aimed to identify proteins involved in NAFLD development in free fatty acids (FFA)‐induced hepatosteatotic cells and in human liver biopsies. Steatosis was induced by incubating a normal human hepatocyte‐derived cell line L‐02 with FFA. Differentially expressed proteins in the steatotic cells were analyzed by two‐dimensional gel electrophoresis‐based proteomics. Involvement of one of the up‐regulated proteins in steatosis was characterized using the RNA interference approach with the steatotic cells. Protein expression levels in liver biopsies of patients with NAFLD were assessed by immunohistochemistry. Proteomic analysis of L‐02 steatotic cells revealed the up‐regulation of ERp57, a condition not previously implicated in NAFLD. Knockdown of ERp57 expression with siRNA significantly reduced fat accumulation in the steatotic cells. ERp57 expression was detected in 16 out of 17 patient biopsies and correlated with inflammation grades or fibrosis stages, while in 5 normal biopsies ERp57 expression was not detectable in hepatocytes. In conclusion, ERp57 was up‐regulated in FFA‐induced steatotic hepatic cells and in NAFLD patient livers and demonstrated steatotic properties in cultured cells. Further investigations are warranted to verify the involvement of ERp57 in NAFLD development. J. Cell. Biochem. 110: 1447–1456, 2010. © 2010 Wiley‐Liss, Inc.
Differential effect of ionizing radiation exposure on multipotent and differentiation‐restricted bone marrow mesenchymal stem cells
Authors: Mussano Federico , Lee Kenneth J. , Zuk Patricia , Tran Lisa , Cacalano Nicholas A. , Jewett Anahid , Carossa Stefano , Nishimura Ichiro
Published: May 2010

Debilitating effects of bone marrow from ionizing radiation exposure has been well established for hematopoietic stem cells; however, radiation toxicity of mesenchymal stem cells (MSCs) has been controversial. The present study addressed if ionizing radiation exposure differently affected bone marrow MSCs with various differentiation commitments. Mouse bone‐marrow‐derived MSCs, D1 cells of early passages (≤5 passages; p5) maintained the complete characteristics of multipotent MSCs, whereas, after ≥45 passages (p45) the differentiation capability of D1 cells became partially restricted. Both p5 and p45 D1 cells were subjected to single dose irradiation by radioactive isotope 137Cs. Radiation treatment impaired cell renewal and differentiation activities of p5 D1 cells; however, p45 D1 cells were less affected. Radiation treatment upregulated both pro‐ and anti‐apoptotic genes of p5 D1 cells in a dose‐dependent manner, potentially resulting in the various apoptosis thresholds. It was found that constitutive as well as radiation‐induced phosphorylation levels of histone H2AX was significantly higher in p45 D1 cells than in p5 D1 cells. The increased repair activity of DNA double‐strand breakage may play a role for p45 D1 cells to exhibit the relative radioresistance. In conclusion, the radiation toxicity predominantly affecting multipotent MSCs may occur at unexpectedly low doses, which may, in part, contribute to the catabolic pathology of bone tissue. J. Cell. Biochem. 111: 322–332, 2010. © 2010 Wiley‐Liss, Inc.
Temporal and spatial expression of osteoactivin during fracture repair
Authors: Abdelmagid Samir M. , Barbe Mary F. , Hadjiargyrou Michael , Owen Thomas A. , Razmpour Roshanak , Rehman Saqib , Popoff Steven N. , Safadi Fayez F.
Published: May 2010

We previously identified osteoactivin (OA) as a novel secreted osteogenic factor with high expression in developing long bones and calvaria, and that stimulates osteoblast differentiation and matrix mineralization in vitro. In this study, we report on OA mRNA and protein expression in intact long bone and growth plate, and in fracture calluses collected at several time points up to 21 days post‐fracture (PF). OA mRNA and protein were highly expressed in osteoblasts localized in the metaphysis of intact tibia, and in hypertrophic chondrocytes localized in growth plate, findings assessed by in situ hybridization and immunohistochemistry, respectively. Using a rat fracture model, Northern blot analysis showed that expression of OA mRNA was significantly higher in day‐3 and day‐10 PF calluses than in intact rat femurs. Using in situ hybridization, we examined OA mRNA expression during fracture healing and found that OA was temporally regulated, with positive signals seen as early as day‐3 PF, reaching a maximal intensity at day‐10 PF, and finally declining at day‐21 PF. At day‐5 PF, which correlates with chondrogenesis, OA mRNA levels were significantly higher in the soft callus than in intact femurs. Similarly, we detected high OA protein immunoexpression throughout the reparative phase of the hard callus compared to intact femurs. Interestingly, the secreted OA protein was also detected within the newly made cartilage matrix and osteoid tissue. Taken together, these results suggest the possibility that OA plays an important role in bone formation and serves as a positive regulator of fracture healing. J. Cell. Biochem. 111: 295–309, 2010. © 2010 Wiley‐Liss, Inc.
Anti‐invasion effect of rosmarinic acid via the extracellular signal‐regulated kinase and oxidation–reduction pathway in Ls174‐T cells
Authors: Xu Yichun , Xu Guanglin , Liu Li , Xu Desheng , Liu Jianwen
Published: May 2010

Rosmarinic acid is a major phenylpropanoid isolated fromPrunella vulgarisL., which is a composition of herbal tea for centuries in China. However, the anti‐invasion activity on Ls174‐T human colon carcinoma cells has not been studied. In this study, we investigated the anti‐metastasis functions according to wound healing assay, adhesion assay, and Transwell assay and found that rosmarinic acid could inhibit migration, adhesion, and invasion dose‐dependently. Rosmarinic acid also could decrease the level of reactive oxygen species by enhancing the level of reduced glutathione hormone. In addition, rosmarinic acid repressed the activity and expression of matrix metalloproteinase‐2,9. According to Western blot and quantitative real‐time PCR assay, rosmarinic acid may inhibit metastasis from colorectal carcinoma mainly via the pathway of extracellular signal‐regulated kinase. In animal experiment, intraperitoneal administration of 2 mg of rosmarinic acid reduced weight of tumors and the number of lung nodules significantly compared with those of control group. Therefore, these results demonstrated that rosmarinic acid can effectively inhibit tumor metastasis in vitro and in vivo. J. Cell. Biochem. 111: 370–379, 2010. © 2010 Wiley‐Liss, Inc.
Identification of phospholipase C gamma1 as a protein tyrosine phosphatase mu substrate that regulates cell migration
Authors: Phillips‐Mason Polly J. , Kaur Harpreet , Burden‐Gulley Susan M. , Craig Sonya E.L. , Brady‐Kalnay Susann M.
Published: May 2010

The receptor protein tyrosine phosphatase PTPµ has a cell‐adhesion molecule‐like extracellular segment and a catalytically active intracellular segment. This structure gives PTPµ the ability to transduce signals in response to cell–cell adhesion. Full‐length PTPµ is down‐regulated in glioma cells by proteolysis which is linked to increased migration of these cells in the brain. To gain insight into the substrates PTPµ may be dephosphorylating to suppress glioma cell migration, we used a substrate trapping method to identify PTPµ substrates in tumor cell lines. We identified both PKCδ and PLCγ1 as PTPµ substrates. As PLCγ1 activation is linked to increased invasion of cancer cells, we set out to determine whether PTPµ may be upstream of PLCγ1 in regulating glioma cell migration. We conducted brain slice assays using U87‐MG human glioma cells in which PTPµ expression was reduced by shRNA to induce migration. Treatment of the same cells with PTPµ shRNA and a PLCγ1 inhibitor prevented migration of the cells within the brain slice. These data suggest that PLCγ1 is downstream of PTPµ and that dephosphorylation of PLCγ1 is likely to be a major pathway through which PTPµ suppresses glioma cell migration. J. Cell. Biochem. 112: 39–48, 2011. © 2010 Wiley‐Liss, Inc.
Cytokine Stimulation Promotes Increased Glucose Uptake Via Translocation at the Plasma Membrane of GLUT1 in HEK293 Cells
Authors: Zambrano Angara , Jara Evelyn , Murgas Paola , Jara Clara , Castro Maite A. , Angulo Constanza , Concha Ilona I.
Published: May 2010

Interleukin‐3 (IL‐3) and granulocyte/macrophage colony‐stimulating factor (GM‐CSF) are two of the best‐characterized cell survival factors in hematopoietic cells; these factors induce an increase in Akt activity in multiple cell lines, a process thought to be involved in cellular survival. It is known that growth factors require sustained glucose metabolism to promote cell survival. It has been determined that IL‐3 and GM‐CSF signal for increased glucose uptake in hematopoietic cells. Interestingly, receptors for IL‐3 and GM‐CSF are present in several non‐hematopoietic cell types but their roles in these cells have been poorly described. In this study, we demonstrated the expression of IL‐3 and GM‐CSF receptors in HEK293 cells and analyzed their effect on glucose uptake. In these cells, both IL‐3 and GM‐CSF, increased glucose uptake. The results indicated that this increase involves the subcellular redistribution of GLUT1, affecting glucose transporter levels at the cell surface in HEK293 cells. Also the data directly demonstrates that the PI 3‐kinase/Akt pathway is an important mediator of this process. Altogether these results show a role for non‐insulin growth factors in the regulation of GLUT1 trafficking that has not yet been directly determined in non‐hematopoietic cells. J. Cell. Biochem. 110: 1471–1480, 2010. © 2010 Wiley‐Liss, Inc.
DNA‐backbone radio resistivity induced by spin blockade effect
Authors: Abolfath Ramin M. , Brabec Thomas
Published: May 2010

Coherent control of OH‐free radicals interacting with the spin‐triplet state of a DNA molecule is investigated. A model Hamiltonian for molecular spin singlet‐triplet resonance is developed. We illustrate that the spin‐triplet state in DNA molecules can be efficiently populated, as the spin‐injection rate can be tuned to be orders of magnitudes greater than the decay rate due to small spin‐orbit coupling in organic molecules. Owing to the nano‐second life‐time of OH free radicals, a non‐equilibrium free energy barrier induced by the injected spin triplet state that lasts approximately longer than one‐micro second in room temperature can efficiently block the initial Hydrogen abstraction and DNA damage. For a direct demonstration of the spin‐blockade effect, a molecular simulation based on an ab‐initio Car‐Parrinello molecular dynamics is deployed. © 2010 Wiley Periodicals, Inc. J Comput Chem 2010
Computing ensembles of transitions from stable states: Dynamic importance sampling
Authors: Perilla Juan R. , Beckstein Oliver , Denning Elizabeth J. , Woolf Thomas B.
Published: May 2010

There is an increasing dataset of solved biomolecular structures in more than one conformation and increasing evidence that large‐scale conformational change is critical for biomolecular function. In this article, we present our implementation of a dynamic importance sampling (DIMS) algorithm that is directed toward improving our understanding of important intermediate states between experimentally defined starting and ending points. This complements traditional molecular dynamics methods where most of the sampling time is spent in the stable free energy wells defined by these initial and final points. As such, the algorithm creates a candidate set of transitions that provide insights for the much slower and probably most important, functionally relevant degrees of freedom. The method is implemented in the program CHARMM and is tested on six systems of growing size and complexity. These systems, the folding of Protein A and of Protein G, the conformational changes in the calcium sensor S100A6, the glucose–galactose‐binding protein, maltodextrin, and lactoferrin, are also compared against other approaches that have been suggested in the literature. The results suggest good sampling on a diverse set of intermediates for all six systems with an ability to control the bias and thus to sample distributions of trajectories for the analysis of intermediate states. © 2010 Wiley Periodicals, Inc. J Comput Chem, 2011
Expression of GFAT1 and OGT in podocytes: Transport of glucosamine and the implications for glucose uptake into these cells
Authors: Rogacka Dorota , Piwkowska Agnieszka , Jankowski Maciej , Kocbuch Katarzyna , Dominiczak Marek H. , Stępiński Jan K. , Angielski Stefan
Published: May 2010

Glutamine:fructose‐6‐phosphate amidotransferase (GFAT) and N‐acetylglucosaminyltransferase (OGT) participate in glucosamine (GlcN) production and its utilization in O‐glycosylation, one of key post‐translational modifications of nuclear and cytoplasmic proteins. For this purpose, cells require a high rate of intracellular production of GlcN and/or significant GlcN delivery. We studied the expression of GFAT1 and OGT and measured uptake of glucose and GlcN in cultured rat podocytes, the main cellular component of glomerular filtration barrier. RT‐PCR revealed the presence of both GFAT1 and OGT mRNA. Immunofluorescence of GFAT1 has shown staining signal diffused within the cytoplasm of the cell body and processes. However, OGT was distinctly visible around the nucleus and, in diffuse form, within the cytoplasm of cell bodies and processes. Glucose was transported (1.3 ± 0.2 nmol/min/mg protein) mainly by facilitative transporter systems whilst GlcN uptake (1.1 ± 0.2 nmol/min/mg protein) in a significant part, involved a sodium‐dependent transporter. There was interplay between glucose and GlcN uptake. In the presence of GlcN (50 µM), the rate of glucose uptake decreased by about 50%. The rate of GlcN uptake decreased by 28% in the presence of 5.6 mM glucose. Our results suggest that cultured podocytes possess limited ability to synthesize GlcN internally and therefore may need to receive GlcN from the extracellular environment. J. Cell. Physiol. 225: 577–584, 2010. © 2010 Wiley‐Liss, Inc.
Presence of a functional receptor for GLP‐1 in osteoblastic cells, independent of the cAMP‐linked GLP‐1 receptor
Authors: Nuche‐Berenguer Bernardo , Portal‐Núñez Sergio , Moreno Paola , González Nieves , Acitores Alicia , López‐Herradón Ana , Esbrit Pedro , Valverde Isabel , Villanueva‐Peñacarrillo María L.
Published: May 2010

Glucagon‐like peptide 1 (GLP‐1) controls glucose metabolism in extrapancreatic tissues through receptors other than the pancreatic cAMP‐linked GLP‐1 receptor; also, GLP‐1 induces an insulin‐ and PTH‐independent bone anabolic action in insulin‐resistant and type‐2 diabetic rats. Here we searched for the presence and characteristics of GLP‐1 receptors in osteoblastic MC3T3‐E1 cells. [125I]‐GLP‐1 specific binding to MC3T3‐E1 cells was time‐ and temperature‐dependent, reaching maximal value at 30 min at 25°C; in these conditions, [125I]‐GLP‐1 binding was dissociable, and displaced by GLP‐1, partially by GLP‐2, but not by exendin‐4 (Ex‐4), exendin‐9 (Ex‐9), glucagon or insulin; Scatchard analysis of the unlabeled GLP‐1 data showed high and low affinity binding sites; cross‐linking of GLP‐1 binding revealed an estimated 70 kDa band, almost undetectable in the presence of 10−6 M GLP‐1. GLP‐1, Ex‐9, insulin or glucagon failed to modify cellular cAMP content, while GLP‐2 and Ex‐4 increased it. However, GLP‐1 induced an immediate hydrolysis of glycosylphosphatidylinositols (GPIs) generating short‐lived inositolphosphoglycans (IPGs), and an increase in phosphatidylinositol‐3 kinase (PI3K) and mitogen activated protein kinase (MAPK) activities; Ex‐4 also affected GPIs, but its action was delayed with respect to that of GLP‐1. This incretin was found to decrease Runx2 but increased osteocalcin gene expression, without affecting that of osteoprotegerin or the canonical Wnt pathway activity in MC3T3‐E1 cells which do not express the pancreatic GLP‐1 receptor. Our data demonstrate for the first time that GLP‐1 can directly and functionally interact with osteoblastic cells, possibly through a GPI/IPG‐coupled receptor. J. Cell. Physiol. 225: 585–592, 2010. © 2010 Wiley‐Liss, Inc.
1α,25‐(OH)2D3 acts in the early phase of osteoblast differentiation to enhance mineralization via accelerated production of mature matrix vesicles
Authors: Woeckel V.J. , Alves R.D.A.M. , Swagemakers S.M.A. , Eijken M. , Chiba H. , van der Eerden B.C.J. , van Leeuwen J.P.T.M.
Published: May 2010

1α,25‐dihydroxyitamin D3 (1,25D3) deficiency leads to impaired bone mineralization. We used the human pre‐osteoblastic cell line SV‐HFO, which forms within 19 days of culture an extracellular matrix that starts to mineralize around day 12, to examine the mechanism by which 1,25D3 regulates osteoblasts and directly stimulates mineralization. Time phase studies showed that 1,25D3 treatment prior to the onset of mineralization, rather than during mineralization led to accelerated and enhanced mineralization. This is supported by the observation of unaltered stimulation by 1,25D3 even when osteoblasts were devitalized just prior to onset of mineralization and after 1,25D3 treatment. Gene Chip expression profiling identified the pre‐mineralization and mineralization phase as two strongly distinctive transcriptional periods with only 0.6% overlap of genes regulated by 1,25D3. In neither phase 1,25D3 significantly altered expression of extracellular matrix genes. 1,25D3 significantly accelerated the production of mature matrix vesicles (MVs) in the pre‐mineralization. Duration rather than timing determined the extent of the 1,25D3 effect. We propose the concept that besides indirect effects via intestinal calcium uptake 1,25D3 directly accelerates osteoblast‐mediated mineralization via increased production of mature MVs in the period prior to mineralization. The accelerated deposition of mature MVs leads to an earlier onset and higher rate of mineralization. These effects are independent of changes in extracellular matrix protein composition. These data on 1,25D3, mineralization, and MV biology add new insights into the role of 1,25D3 in bone metabolism and emphasize the importance of MVs in bone and maintaining bone health and strength by optimal mineralization status. J. Cell. Physiol. 225: 593–600, 2010. © 2010 Wiley‐Liss, Inc.
Voltage‐dependent κ‐opioid modulation of action potential waveform‐elicited calcium currents in neurohypophysial terminals
Authors: Velázquez‐Marrero Cristina M. , Marrero Héctor G. , Lemos José R.
Published: May 2010

Release of neurotransmitter is activated by the influx of calcium. Inhibition of Ca2+ channels results in less calcium influx into the terminals and presumably a reduction in transmitter release. In the neurohypophysis (NH), Ca2+ channel kinetics, and the associated Ca2+ influx, is primarily controlled by membrane voltage and can be modulated, in a voltage‐dependent manner, by G‐protein subunits interacting with voltage‐gated calcium channels (VGCCs). In this series of experiments we test whether the κ‐ and µ‐opioid inhibition of Ca2+ currents in NH terminals is voltage‐dependent. Voltage‐dependent relief of G‐protein inhibition of VGCC can be achieved with either a depolarizing square pre‐pulse or by action potential waveforms. Both protocols were tested in the presence and absence of opioid agonists targeting the κ‐ and µ‐receptors in neurohypophysial terminals. The κ‐opioid VGCC inhibition is relieved by such pre‐pulses, suggesting that this receptor is involved in a voltage‐dependent membrane delimited pathway. In contrast, µ‐opioid inhibition of VGCC is not relieved by such pre‐pulses, indicating a voltage‐independent diffusible second‐messenger signaling pathway. Furthermore, relief of κ‐opioid inhibition during a physiologic action potential (AP) burst stimulation indicates the possibility of activity‐dependent modulation in vivo. Differences in the facilitation of Ca2+ channels due to specific G‐protein modulation during a burst of APs may contribute to the fine‐tuning of Ca2+‐dependent neuropeptide release in other CNS terminals, as well. J. Cell. Physiol. 225: 223–232, 2010. © 2010 Wiley‐Liss, Inc.
Overexpression of fucosyltransferase IV promotes A431 cell proliferation through activating MAPK and PI3K/Akt signaling pathways
Authors: Yang Xue‐Song , Liu Shuai , Liu Yue‐Jian , Liu Ji‐Wei , Liu Ting‐Jiao , Wang Xiao‐Qi , Yan Qiu
Published: May 2010

Lewis Y (LeY) is a carbohydrate tumor‐asssociated antigen. The majority of cancer cells derived from epithelial tissue express LeY type difucosylated oligosaccharide. Fucosyltransferase IV (FUT4) is an essential enzyme that catalyzes the synthesis of LeY oligosaccharide. Our previous studies have shown that FUT4 overexpression promotes A431 cell proliferation, but the mechanism is still largely unknown. Herein, we investigated the role of the mitogen‐activated protein kinases (MAPKs) and phosphoinositide‐3 kinase (PI3K)/Akt signaling pathways on FUT4‐induced cell proliferation. Results show that overexpression of FUT4 increases the phosphorylation of ERK1/2, p38 MAPK, and PI3K/Akt. Inhibitors of PI3K (LY294002 and Wortmannin) prevented the phosphorylation of ERK1/2, p38 MAPK, and Akt PI3K). Moreover, phosphorylation of Akt is abolished by inhibitors of ERK1/2 (PD98059) and p38 MAPK (SB203580). These data suggested that FUT4 not only activates MAPK and PI3K/Akt signals, but also promotes the crosstalk among these signaling pathways. In addition, FUT4‐induced stimulation of cell proliferation correlates with increased cell cycle progression by promoting cells into S‐phase. The mechanism involves in increased expression of cyclin D1, cyclin E, CDK 2, CDK 4, and pRb, and decreased level of cyclin‐dependent kinases inhibitors p21 and p27, which are blocked by the inhibitors of upstream signal molecules, MAPK and PI3K/Akt. In conclusion, these studies suggest that FUT4 regulates A431 cell growth through controlling cell cycle progression via MAPK and PI3K/Akt signaling pathways. J. Cell. Physiol. 225: 612–619, 2010. © 2010 Wiley‐Liss, Inc.
The culture and transplantation of human limbal stem cells
Authors: Ahmad Sajjad , Osei‐Bempong Charles , Dana Reza , Jurkunas Ula
Published: May 2010

The cornea is the clear front of the eye and its surface is composed of an epithelium. This is renewed by stem cells located at the limbus, which encircles the periphery of the cornea. These limbal stem cells become lost or deficient in the blinding disease of limbal stem cell deficiency. In this review article, we discuss the historical perspective in managing limbal stem cell deficiency as well as describing the more contemporary treatment options, and in particular the culture and transplantation of human limbal stem cells. This treatment was first proposed 13 years ago and many case series have been presented to date showing promising outcomes of this technique. However, challenges still remain in treating the debilitating disease of limbal stem cell deficiency. Here we discuss some of the questions, which remain to be answered in this field. J. Cell. Physiol. 225: 15–19, 2010. © 2010 Wiley‐Liss, Inc.
B‐type natriuretic peptide and extracellular matrix protein interactions in human cardiac fibroblasts
Authors: Huntley Brenda K. , Ichiki Tomoko , Sangaralingham S. Jeson , Chen Horng H. , Burnett John C.
Published: May 2010

Cardiac fibroblasts (CFs) regulate myocardial remodeling by proliferating, differentiating, and secreting extracellular matrix (ECM) proteins. B‐type natriuretic peptide (BNP) is anti‐fibrotic, inhibits collagen production, augments matrix metalloproteinases, and suppresses CF proliferation. Recently, we demonstrated that the ECM protein fibronectin (FN) augmented production of BNP's second messenger, 3′, 5′ cyclic guanosine monophosphate (cGMP) in CFs, supporting crosstalk between FN, BNP, and its receptor, natriuretic peptide receptor A (NPR‐A). Here, we address the specificity of FN to augment cGMP generation by investigating other matrix proteins, including collagen IV which contains RGD motifs and collagen I and poly‐L‐lysine, which have no RGD domain. Collagen IV showed increased cGMP generation to BNP similar to FN. Collagen I and poly‐L‐lysine had no effect. As FN also interacts with integrins, we then examined the effect of integrin receptor antibody blockade on BNP‐mediated cGMP production. On FN plates, antibodies blocking RGD‐binding domains of several integrin subtypes had little effect, while a non‐RGD domain interfering integrin αvβ3 antibody augmented cGMP production. Further, on uncoated plates, integrin αvβ3 blockade continued to potentiate the BNP/cGMP response. These studies suggest that both RGD containing ECM proteins and integrins may interact with BNP/NPR‐A to modulate cGMP generation. J. Cell. Physiol. 225: 251–255, 2010. © 2010 Wiley‐Liss, Inc.
Link between high‐affinity adenosine concentrative nucleoside transporter‐2 (CNT2) and energy metabolism in intestinal and liver parenchymal cells
Authors: Huber‐Ruano Isabel , Pinilla‐Macua Itziar , Torres Gonzalo , Casado F. Javier , Pastor‐Anglada Marçal
Published: May 2010

Concentrative nucleoside transporter 2 (CNT2) is a high‐affinity adenosine transporter that may play physiological roles beyond nucleoside salvage. Previous reports relate CNT2 function to modulation of purinergic signaling and energy metabolism in intestinal and liver parenchymal cells (Duflot et al., 2004, Mol Cell Biol 24:2710–2719; Aymerich et al., 2006, J Cell Sci 119:1612–1621). In the present study, to further examine the link between CNT2 and energy metabolism, CNT2 protein partners were identified using the bacterial two‐hybrid and GST pull‐down approaches. The N‐terminal segment of CNT2 was used as bait, since proteins lacking this domain display impaired plasma membrane insertion and intracellular retention. Glucose‐regulated protein 58 (GRP58) was identified as a potential rCNT2 partner in pull‐down experiments. Two‐hybrid screening performed against a liver human cDNA library led to the identification of aldolase B as another hCNT2 partner. Aldolase B‐RFP and endogenous GRP58 separately co‐localized with CNT2 in HeLa cells transfected with YFPrCNT2. CNT2 interaction with GRP58 was validated using co‐immunoprecipitation experiments. In HeLa cells, fluorescence resonance energy transfer (FRET) efficiency increased upon fructose addition, consistent with a transient interaction between aldolase B and the transporter. The physiological basis for in vivo interactions was derived from experiments in which GRP58 was inhibited or overexpressed and aldolase B activity stimulated towards glycolysis. GRP58 appeared to be a negative effector of CNT2 function, whereas aldolase B flux modulated CNT2 activity via a mechanism involving acquisition of higher affinity for its substrates. These findings support the theory that CNT2 plays roles other than salvage and establishes links with energy metabolism. J. Cell. Physiol. 225: 620–630, 2010. © 2010 Wiley‐Liss, Inc.
Late glacial to Holocene planktic foraminifera bioevents and climatic record in the South Adriatic Sea
Authors: Siani Giuseppe , Paterne Martine , Colin Christophe
Published: May 2010

Quantitative analyses of planktic foraminifera, sea‐surface temperatures (SSTs), oxygen isotope and radiocarbon dating from a deep‐sea core recovered in the South Adriatic Sea have been used to reconstruct a subcentennial climatic and biochronological record since the late glacial (the last 24 cal. ka BP). The identification and relative abundance of 25 species of planktic foraminifera along the core have provided a continuous record of the faunal changes over this time interval. These results have permitted the establishment of 10 biozones in the South Adriatic Sea based on the appearance and/or disappearance of the main specific taxa, from peaks of abundance and/or by modification in marine conditions. The robust chronology of the South Adriatic core allowed correlation of SST estimates to the GISP2 ice core record, indicating that the main climate changes recorded in Greenland ice cores over the last 24 ka are recorded and globally synchronous with those observed in the South Adriatic Sea. This finding further allows comparison of the planktic foraminifera record with the event stratigraphic scheme proposed by the INTIMATE group in order to better identify the relationship between past climatic changes and the response of microfaunal assemblages in the South Adriatic. Copyright © 2010 John Wiley & Sons, Ltd.
Response of testate amoeba assemblages to environmental and climatic changes during the Lateglacial–Holocene transition at Lake Lautrey (Jura Mountains, eastern France)
Authors: Wall Adeline A. J. , Magny Michel , Mitchell Edward A. D. , Vannière Boris , Gilbert Daniel
Published: May 2010

We tested the response of lacustrine testate amoebae (thecamoebians) to climate and environmental changes for the Lateglacial–Holocene transition. The palaeoenvironmental history of the study site (Lake Lautrey, Jura Mountains, eastern France) was previously established based on high‐resolution multi‐proxy studies of the same core. The present study is characterised by a high taxonomic resolution (54 taxa), inclusion of small species (down to 25 µm) and high total counts (>500 individuals per sample on average). Changes in the composition of testate amoeba assemblages (dominant species and assemblage structure), as well as in the accumulation rate (tests cm−2 a−1), corresponded to major climatic phases (i.e. Oldest Dryas, Bølling–Allerød Interstadial, Younger Dryas, Preboreal) as well as changes in organic matter inputs. Furthermore, decreases in the accumulation rate characterised minor short‐lived cooling events, such as Older Dryas event or Gerzensee oscillation. However, the Preboreal oscillation, which was well registered by other proxies at Lake Lautrey, could not be recognised in the testate amoeba record. This work demonstrates that lacustrine testate amoebae can be used for palaeoclimatic and palaeoecological reconstructions. Nevertheless, a better understanding of the relation between climate, organic matter and lacustrine testate amoebae requires further high‐resolution studies based on multi‐proxy approaches and the development of appropriate modern analogues. Copyright © 2010 John Wiley & Sons, Ltd.
Chironomid‐inferred late‐glacial summer air temperatures from Lough Nadourcan, Co. Donegal, Ireland
Authors: Watson Jenny E. , Brooks Stephen J. , Whitehouse Nicki J. , Reimer Paula J. , Birks H. John B. , Turney Chris
Published: May 2010

Western Ireland, located adjacent to the North Atlantic, and with a strongly oceanic climate, is potentially sensitive to rapid and extreme climate change. We present the first high‐resolution chironomid‐inferred mean July temperature reconstruction for Ireland, spanning the late‐glacial and early Holocene (LGIT, 15–10 ka BP). The reconstruction suggests an initial rapid warming followed by a short cool phase early in the interstadial. During the interstadial there are oscillations in the inferred temperatures which may relate to Greenland Interstadial events GI‐1a–e. The temperature decrease into the stadial occurs in two stages. This two‐stage drop can also be seen in other late‐glacial chironomid‐inferred temperature records from the British Isles. A stepped rise in temperatures into the Holocene, consistent with present‐day temperatures in Donegal, is inferred. The results show strong similarities with previously published LGIT chironomid‐inferred temperature reconstructions, and with the NGRIP oxygen‐isotope curve, which indicates that the oscillations observed in the NGRIP record are of hemispherical significance. The results also highlight the influence of the North Atlantic on the Irish climate throughout the LGIT. Copyright © 2010 John Wiley & Sons, Ltd.
A forest history for the last 10 900 years at the site Storasjö, southern Sweden: implications from beetle assemblages
Authors: Olsson Fredrik , Lemdahl Geoffrey
Published: May 2010

A continuous Holocene beetle record from southern Sweden is presented and compared with results from a previous study, at Stavsåkra. The comparison reveals both regional trends and local differences. The early Holocene, ca. 8900–7500 BC, was characterised by open, pine‐dominated woodlands at both sites. Fire and grazing seem to have maintained the open structure and favoured the established vegetation and fauna. During the middle Holocene, ca. 7500– 900 BC, the woodlands grew denser. There are no indications of grazing at the sites during this time period, whereas fires were frequent. The late Holocene, ca. 900 BC to the present, was characterised by a progressive opening up of the woodlands, mainly by land use such as grazing and clearance by fire. This process started ca. 2400 BC at Stavsåkra, where there were already settlements during the late Neolithic. At Storasjö, which probably was mainly used as outland until medieval times, the woodlands gradually opened up. Heather‐dominated heathland formed between ca. 900 and 800 BC at both sites. The two studies confirm that insect data contribute unique information concerning changes in structures and disturbances in forest ecosystems on long timescales. Copyright © 2010 John Wiley & Sons, Ltd.
Vegetation responses to climatic variability in the Swiss Southern Alps during the Misox event at the early–mid Holocene transition
Authors: Valsecchi Verushka , Tinner Willy
Published: May 2010

Sedimentary pollen, charcoal and plant macrofossil analyses with high resolution and precision suggest a strong shift in vegetation composition during the early to mid‐Holocene transition in the upper mountain belt. At Piano mire (1439 m above sea level (a.s.l.), Ticino, Switzerland) forests were dominated by Abies alba during the early Holocene (prior to ca. 8000 cal. a BP). Abrupt collapses of A. alba at ca. 7800–7400 cal. a BP enabled the expansion of the light‐demanding pioneer Betula. Afterwards A. alba populations regained their previous abundance in the forests. Within the dating uncertainties of our record we assume that a unique combination of wet and cold years between 8400 and 7500 cal. a BP led to repeated lethal disadvantages for Abies. Our record of Abies oscillations is in good biostratigraphic agreement with the record that has been used to define the Misox cold event (Pian di Signano, 1540 m a.s.l.), which has been previously correlated with the 8200 cal. a BP event. Given the age estimates of the Abies collapses in our well‐dated record, our results suggest that additional efforts are needed to understand the linkage between the Misox and the 8200 cal. a BP event. They imply a high sensitivity of mountain vegetation far below the tree line (∼800 m) to Holocene climatic changes of about 2°C in annual air temperature. Copyright © 2010 John Wiley & Sons, Ltd.
Differential gene expression profile of MAGE family in taiwanese patients with colorectal cancer
Authors: Chung Fu‐Yen , Cheng Tian‐Lu , Chang Hui‐Jen , Chiu Hua‐Hsien , Huang Ming‐Yii , Chang Ming‐Sung , Chen Chung‐Chi , Yang Ming‐Je , Wang Jaw‐Yuan , Lin Shiu‐Ru
Published: May 2010

Background

The melanoma‐associated antigen (MAGE) gene family consists of different expression patterns in various tumor types. They are considered tumor‐specific antigens and are ideal targets for cancer immunotherapy. The purpose of this study is to identify the expression profiles of the MAGE family genes in Taiwanese colorectal cancer patients.

Methods

In this study, a well‐constructed chip array platform was used to analyze the expression of the MAGE family genes of 100 colorectal cancer tissues. Statistical analysis of the experimental results and patients' clinical manifestations were also conducted.

Results

The results showed MAGE‐A2 (87%), ‐A7 (83%), ‐A8 (75%), ‐A12 (71%), ‐B2 (75%), ‐B3 (79%), ‐D2 (75%), ‐F1 (79%), and ‐H1 (70%) were significantly overexpressed genes in colorectal cancer tissues. MAGE‐A2 was the most highly overexpressed gene among the MAGE family. MAGE‐B3 gene expression is statistically correlated with tumor size, lymph node, and UICC stage. In addition, the overexpression of MAGE‐D2 and ‐H1 genes are statistically correlated to the tumor size and depth, respectively (P < 0.05).

Conclusions

This is the first comprehensive report to clarify the differential expression profile of whole MAGE family in CRCs, and it might provide some crucial information about the carcinogenesis and progression in Taiwanese patients with CRC. J. Surg. Oncol. 2010;102:148–153. © 2010 Wiley‐Liss, Inc.
Caspase‐8 polymorphisms and risk of gallbladder cancer in a Northern Indian population
Authors: Srivastava Kshitij , Srivastava Anvesha , Mittal Balraj
Published: May 2010

Caspase‐8 (CASP8) is a key controller of apoptosis, and its deregulation plays an important role in carcinogenesis. To evaluate the role of CASP8 polymorphisms in gallbladder cancer (GBC), we examined the risk associated with three single‐nucleotide polymorphisms (SNPs) in a case–control study in North Indian population. Genotypes and haplotypes of the CASP8 polymorphisms (−652 6N ins/del; rs3834129, Ex13 + 51G > C; rs1045485 and IVS12‐19 G > A; rs3769818) were determined for 230 GBC patients and 230 cancer‐free controls randomly selected from the population. Odds ratio (OR) and 95% confidence interval (95% CI) were calculated in multivariate logistic regression analysis for the association of individual SNPs and haplotypes with GBC. Carriers for the “del” allele of rs3834129 SNP were associated with a 0.60‐fold decreased risk for GBC (95% CI = 0.42–0.88; Ptrend = 0.005). In the combined analysis of the three CASP8 variants, we found that the individuals with the diplotypes carrying two copies of the common CASP8 del‐G‐G haplotype had 0.35‐fold reduced risk (95% CI = 0.14–0.85) when compared with the diplotype containing 0–1 copy. The false‐positive report probability (FPRP) approach advocated that these results were noteworthy (FPRP < 0.5). The molecular modeling results of rs1045485 polymorphism indicated that the overall configuration of both wild‐type and polymorphic CASP8 protein were similar, with negligible deviation at the site of the polymorphism itself. In summary, low penetrance variants in CASP8 gene may alter the susceptibility toward GBC. © 2010 Wiley‐Liss, Inc.
Disposition of hop prenylflavonoids in human breast tissue
Authors: Bolca Selin , Li Jinghu , Nikolic Dejan , Roche Nathalie , Blondeel Phillip , Possemiers Sam , De Keukeleire Denis , Bracke Marc , Heyerick Arne , van Breemen Richard B. , Depypere Herman
Published: May 2010

Hop‐derived products may contain xanthohumol (XN), isoxanthohumol (IX), and the potent phytoestrogen 8‐prenylnaringenin (8‐PN). To evaluate the potential health effects of these prenylflavonoids on breast tissue, their concentration, nature of metabolites, and biodistribution were assessed and compared with 17β‐estradiol (E2) exposure. In this dietary intervention study, women were randomly allocated to hop (n=11; 2.04 mg XN, 1.20 mg IX, and 0.1 mg 8‐PNpersupplement) or control (n=10). After a run‐in of ≥4 days, three supplements were taken daily for 5 days preceding an aesthetic breast reduction. Blood and breast biopsies were analyzed using HPLC‐ESI‐MS/MS. Upon hop administration, XN and IX concentrations ranged between 0.72 and 17.65 nmol/L and 3.30 and 31.50 nmol/L, and between 0.26 and 5.14 pmol/g and 1.16 and 83.67 pmol/g in hydrolyzed serum and breast tissue, respectively. 8‐PN however, was only detected in samples of moderate and strong 8‐PN producers (0.43–7.06 nmol/L and 0.78–4.83 pmol/g). Phase I metabolism appeared to be minor (∼10%), whereas extensive glucuronidation was observed (> 90%). Total prenylflavonoids showed a breast adipose/glandular tissue distribution of 38/62 and their derived E2‐equivalents were negligible compared with E2in adipose (384.6±118.8 fmol/g,p=0.009) and glandular (241.6±93.1 fmol/g,p<0.001) tissue, respectively. Consequently, low doses of prenylflavonoids are unlikely to elicit estrogenic responses in breast tissue.
Vitamin D and multiple health outcomes in the Harvard cohorts
Authors: Wei Melissa Y. , Giovannucci Edward L.
Published: May 2010

The hypothesis that vitamin D is inversely associated with multiple health outcomes has been studied in the Harvard cohorts, including the Nurses' Health Study I (n=121 700 female nurses aged 37–64 at baseline in 1984), Nurses' Health Study II (n=116 671 female nurses aged 27–44 years at baseline in 1991), Health Professionals Follow‐up Study (n=51 529 male health professionals aged 40–75 years at baseline in 1986), and Physicians' Health Study (n=22 071 male physicians aged 40–84 years at baseline in 1982). These studies assessed vitamin D through circulating 25‐hydroxyvitamin D, dietary and supplemental intake, predicted 25‐hydroxyvitamin D, and vitamin D receptor polymorphisms. This review summarizes studies of vitamin D and various endpoints considered in these cohorts, including risk of cardiovascular disease, hypertension, elevated plasma C‐peptide, various cancers, bone fractures, and multiple sclerosis. Based on the multiple observed benefits of vitamin D, this article postulates recommendations for vitamin D intake in the US population for reduced incidence of multiple health outcomes.
Characterization of a reverse molding sequence at the mesoscale for in‐mold assembly of revolute joints
Authors: Ananthanarayanan Arvind , Gupta Satyandra K. , Bruck Hugh A.
Published: May 2010

In‐mold assembly and miniature molding have been combined to realize miniature assemblies. Our previous work has shown that for realizing in‐mold assembly at the mesoscale, the molding sequence is the reverse of that used at the macroscale. Moreover, special features are needed in the mold to prevent plastic bending of the pin because of the pressure exerted by the second stage melt. This article presents novel mold designs and computational methods to enable the use of the reverse molding sequence to prevent: (1) plastic bending of the premolded component, and (2) joint jamming during the mesoscale in‐mold assembly process. The computational methods are also validated through experimentation. Results reported in this article show that for making in‐mold assembled revolute joints with the polymer combination of ABS and LDPE, a reversed molding sequence needs to be used for joints sizes less than 1.5 mm. POLYM. ENG. SCI., 50:1843–1852, 2010. © 2010 Society of Plastics Engineers