bluuebunny/crossref_metadata_2025_split huggingface dataset:
Fluorescent labeling of proteins in living cells using the FKBP12 (F36V) tag
Authors: Robers Matt , Pinson Patrick , Leong Louis , Batchelor Robert H. , Gee Kyle R. , Machleidt Thomas 
Published: October 2008

AbstractOver the past decade live cell imaging has become a key technology to monitor and understand the dynamic behavior of proteins in the physiological context of living cells. The visualization of a protein of interest is most commonly achieved by genetically fusing it to green fluorescent protein (GFP) or one of it variants. Considerable effort has been made to develop alternative methods of protein labeling to overcome the intrinsic limitations of fluorescent proteins. In this report we show the optimization of a live cell labeling technology based on the use of a mutant form of FKBP12 (FKBP12(F36V)) in combination with a synthetic high affinity ligand (SLF') that specifically binds to this mutant. It had been previously shown that the use of a fluorescein‐conjugated form of SLF' (5′‐fluorescein‐SLF') allowed the labeling of proteins genetically fused to FKBP‐F36V in living cells. Here we describe the identification of novel fluorescent SLF'dye conjugates that allow specific labeling of FKBP12(F36V) fusion proteins in living cells. To further increase the versatility of this technology we developed a number of technical improvements. We implemented the use of pluronics during the labeling process to facilitate the uptake of the SLF'‐dye conjugates and the use suppression dyes to reduce background signal. Furthermore, the time and dose dependency of labeling was investigated in order to determine optimal labeling conditions. Finally, the specificity of the FKBP12(F36V) labeling technology was extensively validated by morphological analysis using a diverse set of FKBP12(F36V) fusions proteins. In addition we show a number of different application examples, such as translocation assays, the generation of biosensors, and multiplex labeling in combination with different labeling technologies, such as FlAsH or GFP. In summary we show that the FKBP12(F36V)/SLF' labeling technology has a broad range of applications and should prove useful for the study of protein function in living cells. © 2008 International Society for Advancement of Cytometry
Policy profile: a framework for the classification, prioritization and analysis of research for European environmental regulators
Authors: Gardner M. J. , Hunt D. T. E. , Gardner S. , Shackell K. 
Published: October 2008

AbstractThe SKEP ERA‐NET (Scientific Knowledge for Environmental Protection) is a consortium of European public sector organizations funding environmental research. The network is funded by the European Commission's Sixth Framework Programme (FP6). The aim of the network is to facilitate the improvement of science‐into‐policy processes and to support evidence‐based regulation. This paper contains a description of the categorization and review of current environmental research within the network. It also illustrates how a comparison of current activities combined with perceptions of future needs can provide an analysis of research gaps. The importance of an assessment of relative research costs in such a gap analysis is emphasized. Copyright © 2008 John Wiley & Sons, Ltd and ERP Environment.
“This story is right on”: The impact of regulatory fit on narrative engagement and persuasion
Authors: Vaughn Leigh Ann , Hesse Sarah J. , Petkova Zhivka , Trudeau Lindsay 
Published: October 2008

AbstractWhen people read a story, feelings of rightness from regulatory fit (consistency between regulatory state and strategic means) could suggest that the story is “right on” relative to feelings of wrongness from regulatory nonfit. Under these conditions, individuals who are experiencing feelings of rightness should engage more with the narrative and be more persuaded by its implicit messages. Results from two experiments supported these hypotheses. Participants in Experiment 1 were more mentally engaged (transported) by the story when they experienced regulatory fit. We replicated this effect in Experiment 2 and extended it to endorsement of story‐consistent beliefs, an indicator of persuasion via narratives. Additionally, we found that drawing participants' attention to an earlier event as a source of feelings of rightness eliminated the regulatory fit effects on transportation and persuasion, suggesting attribution of feelings of regulatory fit/nonfit to the plausibility of the narrative world. Copyright © 2008 John Wiley & Sons, Ltd.
Interpersonal relationships moderate the effect of faces on person judgments
Authors: Tuk Mirjam A. , Verlegh Peeter W. J. , Smidts Ale , Wigboldus Daniel H. J. 
Published: October 2008

AbstractPrevious research suggests that people form impressions of others based on their facial appearance in a very fast and automatic manner, and this especially holds for trustworthiness. However, as yet, this process has been investigated mostly in a social vacuum without taking interpersonal factors into account. In the current research, we demonstrate that both the relationship context that is salient at the moment of an interaction and the performed behavior are important moderators of the impact of facial cues on impression formation. It is shown that, when the behavior of a person we encounter is ambiguous in terms of trustworthiness, the relationship most salient at that moment is of crucial impact on whether and how we incorporate facial cues communicating (un)trustworthiness in our final evaluations. Ironically, this can result in less positive evaluations of interaction partners with a trustworthy face compared to interaction partners with an untrustworthy face. Implications for research on facial characteristics, trust, and relationship theories are discussed. Copyright © 2008 John Wiley & Sons, Ltd.
European norms and equations for a two dimensional presentation of values as measured with Schwartz's 21‐item portrait values questionnaire
Authors: Verkasalo Markku , Lönnqvist Jan‐Erik , Lipsanen Jari , Helkama Klaus 
Published: October 2008

AbstractThe present study utilizes European Social Survey (ESS) data to introduce European norms and equations for computation of the Self‐Transcendence—Self‐Enhancement and Conservation—Openness to Change value dimensions, as measured with the 21‐item Portrait Values Questionnaire (PVQ). Our analysis of ESS round 1 and 2 data suggest that the two‐dimensional structure and the equations based on this structure are extremely robust. Presenting the two value dimensions besides the 10 basic values offers the advantages of (a) heightened reliability, (b) control of response tendency, and (c) possibility to present results in two‐dimensional space. Copyright © 2008 John Wiley & Sons, Ltd.
Comparative epigenomics of human and mouse mammary tumors
Authors: Demircan Berna , Dyer Lisa M. , Gerace Mallory , Lobenhofer Edward K. , Robertson Keith D. , Brown Kevin D. 
Published: October 2008

AbstractGene silencing by aberrant epigenetic chromatin alteration is a well‐recognized event contributing to tumorigenesis. Although genetically engineered tumor‐prone mouse models have proven a powerful tool in understanding many aspects of carcinogenesis, to date few studies have focused on epigenetic alterations in mouse tumors. To uncover epigenetically silenced tumor suppressor genes (TSGs) in mouse mammary tumor cells, we conducted initial genome‐wide screening by combining the treatment of cultured cells with the DNA demethylating drug 5‐aza‐2′‐deoxycytidine (5‐azadC) and the histone deacetylase inhibitor trichostatin A (TSA) with expression microarray. By conducting this initial screen on EMT6 cells and applying protein function and genomic structure criteria to genes identified as upregulated in response to 5‐azadC/TSA, we were able to identify two characterized breast cancer TSGs (Timp3 and Rprm) and four putative TSGs (Atp1B2, Dusp2, FoxJ1 and Smpd3) silenced in this line. By testing a panel of 10 mouse mammary tumor lines, we determined that each of these genes is commonly hypermethylated, albeit with varying frequency. Furthermore, by examining a panel of human breast tumor lines and primary tumors we observed that the human orthologs of ATP1B2, FOXJ1 and SMPD3 are aberrantly hypermethylated in the human disease whereas DUSP2 was not hypermethylated in primary breast tumors. Finally, we examined hypermethylation of several genes targeted for epigenetic silencing in human breast tumors in our panel of 10 mouse mammary tumor lines. We observed that the orthologs of Cdh1, RarB, Gstp1, RassF1 genes were hypermethylated, whereas neither Dapk1 nor Wif1 were aberrantly methylated in this panel of mouse tumor lines. From this study, we conclude that there is significant, but not absolute, overlap in the epigenome of human and mouse mammary tumors. © 2008 Wiley‐Liss, Inc.
2‐Arachidonoylglycerol elicits neuroprotective effects on excitotoxically lesioned dentate gyrus granule cells via abnormal‐cannabidiol‐sensitive receptors on microglial cells
Authors: Kreutz Susanne , Koch Marco , Böttger Charlotte , Ghadban Chalid , Korf Horst‐Werner , Dehghani Faramarz 
Published: October 2008

AbstractEndocannabinoids like 2‐arachidonoylglycerol (2‐AG) exert neuroprotective effects after brain injuries. According to current concepts, these neuroprotective effects are due to interactions between 2‐AG and cannabinoid (CB)1 receptors on neurons. Moreover, 2‐AG modulates migration and proliferation of microglial cells which are rapidly activated after brain lesion. This effect is mediated via CB2‐ and abnormal‐cannabidiol (abn‐CBD)‐sensitive receptors. In the present study, we investigated whether the abn‐CBD‐sensitive receptor on microglial cells contributes to 2‐AG‐mediated neuroprotection in organotypic hippocampal slice cultures (OHSCs) after excitotoxic lesion induced by NMDA (50 μM) application for 4 h. This lesion caused neuronal damage and accumulation of microglial cells within the granule cell layer. To analyze the role of abn‐CBD‐sensitive receptors for neuroprotection and microglial cell accumulation, two agonists of the abn‐CBD‐sensitive receptor, abn‐CBD or 2‐AG, two antagonists, 1,3‐dimethoxy‐5‐methyl‐2‐[(1R,6R)‐3‐methyl‐6‐(1‐methylethenyl)‐2‐cyclohexen1‐yl]‐benzene (O‐1918) or cannabidiol (CBD), and the CB1 receptor antagonist AM251, were applied to NMDA‐lesioned OHSC. Propidium iodide (PI) labeling was used as a marker of degenerating neurons and isolectin B4 (IB4) as a marker of microglial cells. Application of both, abn‐CBD or 2‐AG to lesioned OHSC significantly decreased the number of IB microglial cells and PI+ neurons in the dentate gyrus. In contrast to AM251, application of O‐1918 or CBD antagonized these effects. When microglial cells were depleted by preincubation of OHSC with the bisphosphonate clodronate (100 μg/mL) for 5 days before excitotoxic lesion, 2‐AG and abn‐CBD lost their neuroprotective effects. We therefore propose that the endocannabinoid 2‐AG exerts its neuroprotective effects via activation of abn‐CBD‐sensitive receptors on microglial cells. © 2008 Wiley‐Liss, Inc.
Depression of parallel and climbing fiber transmission to Bergmann glia is input specific and correlates with increased precision of synaptic transmission
Authors: Balakrishnan Saju , Bellamy Tomas C. 
Published: October 2008

AbstractIn the cerebellar cortex, Bergmann glia enclose the synapses of both parallel and climbing fiber inputs to the Purkinje neuron. The glia express Ca2+‐permeable AMPA receptors, and the GLAST and GLT‐1 classes of glutamate transporter, which are activated by glutamate released during synaptic transmission. We have previously reported that parallel fiber to Bergmann glial transmission in rat cerebellar slices exhibits a form of frequency‐dependent plasticity, namely long‐term depression, following repetitive stimulation at 0.1–1 Hz. Here, we report that this form of plasticity is also present at the climbing fiber input, that climbing and parallel fibers can be depressed independently, that discrete parallel fiber inputs can also be depressed independently, and that depression is maintained when a distributed array of parallel fibers are stimulated (in contrast to several forms of synaptic plasticity at the Purkinje neuron). Depression of glutamate transporter currents does not correlate with a decrease in the stringency with which Purkinje neuron synapses are isolated. Rather, postsynaptic currents in Purkinje neurons decay more rapidly and perisynaptic metabotropic glutamate receptors are activated less effectively after stimulation at 0.2 and 1 Hz, suggesting that depression arises from a decrease in extrasynaptic glutamate concentration and not from impairment of glutamate clearance in and around the synapse. These results indicate that neuron‐glial plasticity is activity dependent, input specific and does not require spillover between adjacent synapses to manifest. They also argue against a withdrawal of the glial sheath from synaptic regions as the putative mechanism of plasticity. © 2008 Wiley‐Liss, Inc.
Quantitative analysis of mitotic Olig2 cells in adult human brain and gliomas: Implications for glioma histogenesis and biology
Authors: Rhee Wootack , Ray Sutapa , Yokoo Hideaki , Hoane Megan E. , Lee Chong C. , Mikheev Andrei M. , Horner Philip J. , Rostomily Robert C. 
Published: October 2008

AbstractThe capacity of adult human glial progenitor cells (AGPs), to proliferate and undergo multipotent differentiation, positions them as ideal candidate cells of origin for human gliomas. To investigate this potential role we identified AGPs as mitotically active Olig2 cells in nonneoplastic adult human brain and gliomas. We conservatively estimated that one in 5,000 human temporal lobe neocortical gray or subcortical white matter cells is mitotic. Extrapolating from a mean Olig2/Mib‐1 labeling index (LI) of 52% and total cell number of 100 billion, we estimated the overall prevalence of mitotic Olig2 AGPs in nonneoplastic human brain parenchyma at 10 million. These data identify a large reservoir of Olig2 AGPs which could be potential targets for human gliomagenesis. The vast majority of mitotic cells in Grade II and Grade III gliomas of all histologic subtypes expressed Olig2 (mean LI 75%) but rarely S100B (LI 0.6%), identifying the Olig2 cell as a distinct contributor to the proliferating cell population of human gliomas of both oligodendroglial and astrocytic lineages. In the most malignant Grade IV glioma, or glioblastoma multiforme (GBM), the prevalence of Olig2/Mib‐1 cells was significantly decreased (24.5%). The significantly lower Olig2/Mib‐1 LI in GBMs suggests that a decrease in the prevalence of Olig2 cells to the total mitotic cell pool accompanies increasing malignancy. The novel framework provided by this quantitative and comparative analysis supports future studies to examine the histogenetic role of Olig2 AGPs in adult gliomas, their potential contribution to the tumor stroma and the molecular role of Olig2 in glioma pathogenesis. © 2008 Wiley‐Liss, Inc.
The giant protein AHNAK involved in morphogenesis and laminin substrate adhesion of myelinating Schwann cells
Authors: Salim Claudio , Boxberg Ysander V. , Alterio Jeanine , Féréol Sophie , Nothias Fatiha 
Published: October 2008

AbstractWithin the nervous system, expression of the intriguing giant protein AHNAK had been reported so far only for blood–brain barrier forming vascular endothelium. In a screen for genes upregulated after spinal cord injury, we recently identified ahnak as being highly expressed by non‐neuronal cells invading the lesion, delimiting the interior surface of cystic cavities in front of barrier‐forming astrocytes. Here, we show for the first time that AHNAK is constitutively expressed in peripheral nervous system, notably by myelinating Schwann cells (SCs), in which we investigated its function. During sciatic nerve development, AHNAK is redistributed from adaxonal toward abaxonal SC compartments in contact with basement membrane. AHNAK labeling on myelinated fibers from adult nerve delineates the so‐called “Cajal bands,” constituting the residual peripheral SC cytoplasm. Its distribution pattern is complementary to that of periaxin, known to be involved in the myelination process. In vitro, nonconfluent cultured primary SCs seeded on laminin express high levels of AHNAK concentrated in their processes, whereas at confluence, AHNAK is downregulated together with laminin receptor dystroglycan. AHNAK silencing by siRNA interference affects SC morphology and laminin‐substrate attachment, as well as expression and distribution of dystroglycan. Thus, our results clearly show the implication of AHNAK in SC adhesion to laminin, probably via targeting of the dystroglycan‐associated receptor complex. These findings are of high interest regarding the importance of SC‐basal lamina interactions for myelination and myelin maintenance, and open up new perspectives for investigations of the molecular mechanisms underlying demyelinating neuropathies. © 2008 Wiley‐Liss, Inc.
Oligodendrocytes and myelination: The role of iron
Authors: Todorich Bozho , Pasquini Juana M. , Garcia Corina I. , Paez Pablo M. , Connor James R. 
Published: October 2008

AbstractIron is an essential trophic factor that is required for oxygen consumption and ATP production. Thus it plays a key role in vital cell functions. Although the brain has a relatively high rate of oxygen consumption compared to other organs, oligodendrocytes are the principal cells in the CNS that stain for iron under normal conditions. The importance of iron in myelin production has been demonstrated by studies showing that decreased availability of iron in the diet is associated with hypomyelination. The timing of iron delivery to oligodendrocytes during development is also important because hypomyelination and the associated neurological sequelae persist long after the systemic iron deficiency has been corrected. Therefore, identifying the molecular roles of iron in oligodendrocyte development and myelin production, and the mechanisms and timing of iron acquisitions are important prerequisites to developing effective therapies for dysmyelinating disorders. It is the purpose of this review to give a comprehensive overview of the existing literature on role of iron in oligodendrocytes and the mechanisms of iron acquisition and intracellular handling. © 2008 Wiley‐Liss, Inc.
ATP induces long‐term potentiation of C‐fiber‐evoked field potentials in spinal dorsal horn: The roles of P2X4 receptors and p38 MAPK in microglia
Authors: Gong Qing‐Juan , Li Yu‐Ying , Xin Wen‐Jun , Zang Ying , Ren Wen‐Jie , Wei Xu‐Hong , Li Yong‐Yong , Zhang Tong , Liu Xian‐Guo 
Published: October 2008

AbstractMany studies have shown that adenosine triphosphate (ATP), as a neurotransmitter, is involved in plastic changes of synaptic transmission in central nervous system. In the present study, we tested whether extracellular ATP can induce long‐term potentiation (LTP) of C‐fiber‐evoked field potentials in spinal dorsal horn. The results showed the following: (1) ATP at a concentration of 0.3 mM induced spinal LTP of C‐fiber‐evoked field potentials, lasting for at least 5 h; (2) spinal application of 2′,3′‐O‐(2,4,6‐trinitrophenyl)adenosine‐5‐triphosphate (TNP‐ATP; an antagonist of P2X1–4 receptors), but not pyridoxal‐phosphate‐6‐azophenyl‐2′,4′‐disulphonic acid (PPADS; an antagonist of P2X1,2,3,5,7 receptors), 30 min before ATP blocked ATP‐induced LTP, indicating that ATP may induce spinal LTP by activation of P2X4 receptors; (3) at 60 min after LTP induction the level of phospho‐p38 mitogen‐activated protein kinase (p‐p38 MAPK) was significantly elevated and at 180 min after LTP the number of P2X4 receptors increased significantly; both p‐p38 and P2X4 receptors were exclusively co‐located with the microglia marker, but not with neuronal or astrocyte marker; (4) spinal application of TNP‐ATP but not PPADS prevented p38 activation; (5) spinal application of SB203580, a p38 MAPK inhibitor, prevented both spinal LTP and the upregulation of P2X4 receptors. The results suggested that ATP may activate p38 MAPK by binding to intrinsic P2X4 receptors in microglia, and subsequently enhance the expression of P2X4 receptors, contributing to spinal LTP. © 2008 Wiley‐Liss, Inc.
Flow cytometric isolation and clonal identification of self-renewing bipotent hepatic progenitor cells in adult mouse liver
Authors: Suzuki Atsushi , Sekiya Sayaka , Onishi Makiko , Oshima Naoko , Kiyonari Hiroshi , Nakauchi Hiromitsu , Taniguchi Hideki 
Published: October 2008

Abstract

The adult liver progenitor cells appear in response to several types of pathological liver injury, especially when hepatocyte replication is blocked. These cells are histologically identified as cells that express cholangiocyte markers and proliferate in the portal area of the hepatic lobule. Although these cells play an important role in liver regeneration, the precise characterization that determines these cells as self-renewing bipotent primitive hepatic cells remains to be shown. Here we attempted to isolate cells that express a cholangiocyte marker from the adult mouse liver and perform single cell-based analysis to examine precisely bilineage differentiation potential and self-renewing capability of these cells. Based on the results of microarray analysis and immunohistochemistry, we used an antibody against CD133 and isolate CD133+ cells via flow cytometry. We then cultured and propagated isolated cells in a single cell culture condition and examined their potential for proliferation and differentiation

in vitro

and

in vivo

. Isolated cells that could form large colonies (LCs) in culture gave rise to both hepatocytes and cholangiocytes as descendants, while maintaining undifferentiated cells by self-renewing cell divisions. The clonogenic progeny of an LC-forming cell is capable of reconstituting hepatic tissues

in vivo

by differentiating into fully functional hepatocytes. Moreover, the deletion of

p53

in isolated LC-forming cells resulted in the formation of tumors with some characteristics of hepatocellular carcinoma and cholangiocarcinoma upon subcutaneous injection into immunodeficient mutant mice. These data provide evidence for the stem cell-like capacity of isolated and clonally cultured CD133+ LC-forming cells.

Conclusion:

Our method for prospectively isolating hepatic progenitor cells from the adult mouse liver will facilitate study of their roles in liver regeneration and carcinogenesis. (Hepatology 2008;48:1964-1978.)
Great toe transplant versus thumb replant for isolated thumb amputation: Critical analysis of functional outcome
Authors: Rosson Gedge D. , Buncke Gregory M. , Buncke Harry J. 
Published: October 2008

AbstractBackground: Thumb replantation following thumb amputation is the standard of care. When replantation is impossible, microneurovascular great toe transplantation is a well‐established option. Methods: A retrospective review was conducted to evaluate functional outcome following isolated thumb replantation or great toe transplantation for thumb reconstruction. From 1974 to 1993, 384 thumb amputations were treated and 110 great toe‐to‐thumb transplantations were performed. Results: Ninety‐one patients with isolated thumb amputation had an 85% survival rate. Failed replants usually resulted from crushing or avulsing injuries. Function of replanted thumbs was better in sharp compared with crush/avulsion injuries. Forty‐three isolated thumb reconstructions had a 93% success rate. Function was comparable with thumb replants from sharp injuries. Interphalangeal motion was significantly better in great toe transplants than in replanted thumbs of the crush/avulsion type. Conclusions: Amputated thumbs should be replanted. When replantation is not possible or unsuccessful, a transplanted great toe functions as well as, or better than, a replanted thumb. © 2008 Wiley‐Liss, Inc. Microsurgery, 2008.