bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inducible activation of IFI 16 results in suppression of telomerase activity, growth suppression and induction of cellular senescence
Authors: Clarke Christopher J.P. , Hii Linda L. , Bolden Jessica E. , Johnstone Ricky W. 
Published: November 2009

AbstractExpression of the human HIN‐200 family member IFI 16 has been reported to suppress cell growth and contribute to the onset of cellular senescence. However the molecular events involved in this process have not been fully characterised. We fused IFI 16 to the estrogen receptor ligand‐binding domain to establish an inducible model for studying the molecular events that cause these phenomena. In cells induced to express the ER‐IFI 16 within the nucleus there was a decrease in cellular proliferation and concomitant growth arrest in the G1 phase of the cell cycle. Unlike previous reports, this did not appear to involve the p53‐p21WAF1/CIP1‐cdk2‐pRb pathway. Following nuclear expression of ER‐IFI 16 we noted senescence‐like morphological changes and expression of senescence‐associated β‐galactosidase in growth arrested cells. Importantly, we also found a marked reduction in telomerase activity in arrested cells compared to controls. Moreover, IFI 16 and hTERT co‐localised within the nucleus and these two proteins physically interacted in vivo and in vitro. Together, these data suggest that IFI 16 may act as an endogenous regulator of telomerase activity and, through its interaction with hTERT, contributes to the inhibition of proliferation and induces a senescence‐like state. J. Cell. Biochem. 109: 103–112, 2010. © 2009 Wiley‐Liss, Inc.
Restricted dead‐end elimination: Protein redesign with a bounded number of residue mutations
Authors: Safi Maria , Lilien Ryan H. 
Published: November 2009

AbstractDead‐end elimination (DEE) has emerged as a powerful structure‐based, conformational search technique enabling computational protein redesign. Given a protein withnmutable residues, the DEE criteria guide the search toward identifying the sequence of amino acids with the global minimum energy conformation (GMEC). This approach does not restrict the number of permitted mutations and allows the identified GMEC to differ from the original sequence in up tonresidues. In practice, redesigns containing a large number of mutations are often problematic when taken into the wet‐lab for creation via site‐directed mutagenesis. The large number of point mutations required for the redesigns makes the process difficult, and increases the risk of major unpredicted and undesirable conformational changes. Preselecting a limited subset of mutable residues is not a satisfactory solution because it is unclear how to select this set before the search has been performed. Therefore, the ideal approach is what we define as the κ‐restricted redesign problem in which any κ of thenresidues are allowed to mutate. We introduce restricted dead‐end elimination (rDEE) as a solution of choice to efficiently identify the GMEC of the restricted redesign (the κGMEC). Whereas existing approaches requiren‐choose‐κ individual runs to identify the κGMEC, the rDEE criteria can perform the redesign in a single search. We derive a number of extensions to rDEE and present a restricted form of the A* conformation search. We also demonstrate a 10‐fold speed‐up of rDEE over traditional DEE approaches on three different experimental systems. © 2009 Wiley Periodicals, Inc. J Comput Chem, 2010
Electronic structure and reactivity of guanylthiourea: A quantum chemical study
Authors: Mehdi Ahmed , Adane Legesse , Patel Dhilon S. , Bharatam Prasad V. 
Published: November 2009

AbstractElectronic structure analysis of guanylthiourea (GTU) and its isomers has been carried out using quantum chemical methods. Two major tautomeric classes (thione and thiol) have been identified on the potential energy (PE) surface. In both the cases conjugation of pi‐electrons and intramolecular H‐bonds have been found to play a stabilizing role. Various isomers of GTU on its PE surface have been analyzed in two different groups (thione and thiol). The interconversion from the most stable thione conformer (GTU‐1) to the most stable thiol conformer (GTU‐t1) was found to take place via bimolecular process which involves protonation at sulfur atom of GTU‐1 followed by subsequent CN bond rotation and deprotonation. The detailed analysis of the protonation has been carried out in gas phase and aqueous phase (using CPMC model). Sulfur atom (S1) was found to be the preferred protonation site (over N4) in GTU‐1 in gas phase whereas N4 was found to be the preferred site of protonation in aqueous medium. The mechanism of S‐alkylation reaction in GTU has also been studied. The formation of alkylated analogs of thiol isomers (alkylated guanylthiourea) is believed to take place via bimolecular process which involves alkyl cation attack at S atom followed by CN bond rotation and deprotonation. The reactive intermediate RS(NH2)CNC(NH2)2+ belongs to the newly identified ⊕N(←L)2 class of species and provides the necessary dynamism for easy conversion of thione to thiol. © 2009 Wiley Periodicals, Inc. J Comput Chem, 2010
Molecular dissection of Otx1 functional domains in the zebrafish embryo
Authors: Bellipanni Gianfranco , Murakami Tohru , Weinberg Eric S. 
Published: November 2009

AbstractOtx proteins are involved in the induction of neurectoderm patterning and morphogenetic movements, leading to the formation of the vertebrate central nervous system. Despite lack of homology of sequence outside the homeodomain, a large body of evidence has shown that the Otx/Otd class of proteins has similar functions in many animal phyla. Thus, characterization of functional domains in proteins of this family would help in understanding how this functional equivalence operates. Our previous analysis using the zebrafish embryo (Bellipanni et al., 2000, Dev Biol 223:339–353), has suggested that induction of cell aggregation is a morphoregulatory role of Otx/Otd factors in embryonic development. We now use the induction of cell aggregation as an in vivo assay to examine the functional requirement for particular domains of the zOtx1 protein. We demonstrate that zOtx1 induces cell aggregation by acting as a transcriptional activator through its C‐terminal region. Further, we show that a region of 37 amino acids in the C‐terminal third of zOtx1 is necessary but not sufficient for this activation potential. The effects of selective deletion of each of the three homeodomain α‐helices of zOtx1 on cell aggregation were also tested. Surprisingly, we find that helix 3, which is required for binding to DNA, is dispensable for stimulation of cell aggregation. Our results suggest that for transcriptional activation of at least one gene in the cell aggregation pathway, zOtx1 need not bind directly to DNA, but does require helix 1 and 2 of its homeodomain to interact with an as yet undefined DNA binding protein. J. Cell. Physiol. 222: 286–293, 2010. © 2009 Wiley‐Liss, Inc.
FEZ1/LZTS1 protein expression in ovarian cancer
Authors: Califano Daniela , Pignata Sandro , Pisano Carmela , Greggi Stefano , Laurelli Giuseppe , Losito Nunzia Simona , Ottaiano Alessandro , Gallipoli Adolfo , Pasquinelli Rosa , De Simone Veronica , Cirombella Roberto , Fusco Alfredo , Chiappetta Gennaro 
Published: November 2009

AbstractThe FEZ1/LZTS1 (FEZ1) gene maps to chromosome 8p22 and is frequently altered in human cancer. FEZ1 has been proposed as a candidate tumour suppressor gene and its loss may contribute to tumour progression. We have analysed the expression of FEZ1 protein in tissues from ovarian carcinomas in relation to clinico‐pathological variables, response to chemotherapy and disease‐free and overall survival. FEZ1 status was assessed by immunohistochemistry. Cytoplasmic staining for FEZ1 protein was absent or drastically reduced in 38% of tumours. FEZ1 protein expression was not related to tumour grade, histotype, disease‐free survival, or overall survival. On the contrary, it was significantly correlated with age and with FIGO stage of disease. This finding indicates that FEZ1 is involved in ovarian carcinogenesis. Moreover, loss of FEZ1 protein significantly predicted a complete treatment response in patients who received taxane‐based chemotherapy. In conclusion, the reduction or loss of FEZ1 protein could be an aid to the clinical management of patients affected by ovarian carcinoma. J. Cell. Physiol. 222: 382–386, 2010. © 2009 Wiley‐Liss, Inc.
Inhibition of PARP activity by PJ‐34 leads to growth impairment and cell death associated with aberrant mitotic pattern and nucleolar actin accumulation in M14 melanoma cell line
Authors: Chevanne Marta , Zampieri Michele , Caldini Riccardo , Rizzo Angela , Ciccarone Fabio , Catizone Angela , D'Angelo Carmen , Guastafierro Tiziana , Biroccio Anna , Reale Anna , Zupi Gabriella , Caiafa Paola 
Published: November 2009

AbstractThe capability of PARP activity inhibitors to prevent DNA damage recovery suggested the use of these drugs as chemo‐ and radio‐sensitisers for cancer therapy. Our research, carried out on cultured human M14 melanoma cells, was aimed to examine if PJ‐34, a potent PARP activity inhibitor of second generation, was per se able to affect the viability of these cancer cells without any DNA damaging agents. Using time‐lapse videomicroscopy, we evidenced that 10 µM PJ‐34 treatment induced severe mitotic defects leading to dramatic reduction of cell proliferation and to cell death. PJ‐34 cytotoxic effect was further confirmed by analysis of cell viability and clonogenic assay. Absence of canonic apoptosis markers allowed us to exclude this kind of cell death. No single and/or double stranded DNA damage was evidenced. Immunofluorescence analysis showed an aberrant mitotic scenario in several cells and subsequent multinucleation suggesting an atypical way for cells to die: the mitotic catastrophe. The detection of aberrant accumulation of polymerised actin inside the nucleolus was noteworthy. Taken toghether, our results demonstrate that, targeting PARP activity by PJ‐34, cancer cell survival is affected independently of DNA damage repair. Two findings are remarkable: (a) cisplatin concentration can be reduced by three quarters if it is followed by treatment with 10 µM PJ‐34 for 24 h to obtain the same citotoxic effect; (b) effects dependent on PJ‐34 treatment are reversible. Our data suggest that, to reduce the harm done to non‐tumour cells during chemotherapy with cisplatin, the latter could be coupled with PJ‐34 treatment. J. Cell. Physiol. 222: 401–410, 2010. © 2009 Wiley‐Liss, Inc.
Characterization of Bombyx embryo segmentation process: expression profiles of engrailed, even‐skipped, caudal, and wnt1/wingless homologues
Authors: Nakao Hajime 
Published: November 2009

AbstractTo gain insight into segmentation processes, the expression at embryonic stages of the silkmoth Bombyx mori homologues of even‐skipped (eve), engrailed (en), caudal (cad), and wnt1/wingless (wg) transcripts were examined by whole mount in situ hybridization. Pair‐rule eve stripes and segmental en and wnt1/wg stripes were generated sequentially from anterior to posterior, confirming the previous results that showed that Bombyx belongs to short‐germ insects. However, unlike in previously described short germ insects, the segmentation of Bombyx occurred without marked germ band elongation: the putative growth zone was expanded compared with previously described short germ insects. This may indicate that Bombyx represents an evolutionarily intermediate state in a transition from short to long germ type. The expressions of cad and wnt1/wg, which are known to be present in the growth zone in short germ insects, initially showed a large median expression domain that, as segmentation proceeded, later retracted to the posterior pole. This is also unique to this insect. Detailed analysis of their relative expressions indicated that wnt1/wg domain retracted faster than the cad domain, and double stain in situ hybridization suggested that the eve stripe appears from cells that have ceased to express wnt1/wg. Another unique aspect of Bombyx embryogenesis is that gastrulation began at later embryonic stage compared with other insects and proceeded slowly from anterior to posterior. On the basis of these results, conserved and divergent aspects of the evolution of insect segmentation mechanisms and germ cell formation are discussed. J. Exp. Zool. (Mol. Dev. Evol.) 314B:224–231, 2010. © 2009 Wiley‐Liss, Inc.
Synthesis and molecular recognition of novel multiimidazole cyclophanes
Authors: Xu Xiao‐Wei , Wang Xin‐Long , Wu Ai‐Ming , Zheng Zhi‐Ming , Yi Mei‐Gui , Xiao Rong 
Published: November 2009

Abstract
magnified image Cyclophanes based on 2,2′‐biimidazole and 2,2′‐bibenzimidazole were synthesized as receptors. UV spectroscopic titration in chloroform at 25°C showed 1:1 complexes between the cyclophanes and the guests, and the binding constants (K) and Gibbs free energy changes (−ΔG0) were calculated according to the modified Benesi‐Hildebrand equation. J. Heterocyclic Chem., 00, 00 (2009).
111In‐labeled KCCYSL peptide as an imaging probe for ErbB‐2‐expressing ovarian carcinomas
Authors: Deutscher Susan L. , Figueroa Said D. , Kumar Senthil R. 
Published: November 2009

AbstractAberrant expression of ErbB‐2, a member of the epidermal growth factor family of receptors, has been implicated in the formation of various malignancies including ovarian cancer. The objective of this study was to determine whether the phage display‐selected ErbB‐2 targeting peptide, KCCYSL, once radiolabeled with 111In would serve as a tumor targeting and single photon emission computed tomography (SPECT/CT) imaging agent in a mouse model of human ovarian carcinoma expressing ErbB‐2. The KCCYSL peptide was synthesized with a chelator 1,4,7,10‐tetra‐azacyclododecane‐N,N′,N″,N′″‐tetraacetic acid (DOTA), and a Gly‐Ser‐Gly (GSG) spacer between DOTA and amino terminus of the peptide and radiolabeled with 111InC13. In vitro cell binding studies indicated that 111In‐DOTA‐GSG‐KCCYSL bound to cultured ovcar‐3 carcinoma cells. Bio‐distribution studies in scid mice bearing human ovcar‐3 tumor xenografts revealed a tumor uptake of 0.50±0.05 percent injected dose per gram (%ID/g) at 1 h, and 0.39±0.1 %ID/g at 2 h. Blocking studies with non‐radiolabeled counterpart indicated a partial inhibition (41%) (P=0.04) in tumor uptake of 111In‐DOTA‐GSG‐KCCYSL. In vivo tumor uptake of 111In‐DOTA‐GSG‐KCCYSL was clearly evident through SPECT/CT images after 2 h post injection. These studies suggest the potential of this peptide as a radiopharmaceutical for imaging of ErbB‐2‐expressing ovarian tumors. Copyright © 2009 John Wiley & Sons, Ltd.
Periaqueductal gray afferents synapse onto dopamine and GABA neurons in the rat ventral tegmental area
Authors: Omelchenko Natalia , Sesack Susan R. 
Published: November 2009

AbstractThe midbrain central gray (periaqueductal gray; PAG) mediates defensive behaviors and is implicated in the rewarding effects of opiate drugs. Projections from the PAG to the ventral tegmental area (VTA) suggest that this region might also regulate behaviors involving motivation and cognition. However, studies have not yet examined the morphological features of PAG axons in the VTA or whether they synapse onto dopamine (DA) or GABA neurons. In this study, we injected anterograde tracers into the rat PAG and used immunoperoxidase to visualize the projections to the VTA. Immunogold‐silver labeling for tyrosine hydroxylase (TH) or GABA was then used to identify the phenotype of innervated cells. Electron microscopic examination of the VTA revealed axons labeled anterogradely from the PAG, including myelinated and unmyelinated fibers and axon varicosities, some of which formed identifiable synapses. Approximately 55% of these synaptic contacts were of the symmetric (presumably inhibitory) type; the rest were asymmetric (presumably excitatory). These findings are consistent with the presence of both GABA and glutamate projection neurons in the PAG. Some PAG axons contained dense‐cored vesicles indicating the presence of neuropeptides in addition to classical neurotransmitters. PAG projections synapsed onto both DA and GABA cells with no obvious selectivity, providing the first anatomical evidence for these direct connections. The results suggest a diverse nature of PAG physiological actions on midbrain neurons. Moreover, as both the VTA and PAG are implicated in the reinforcing actions of opiates, our findings provide a potential substrate for some of the rewarding effects of these drugs. © 2009 Wiley‐Liss, Inc.
Oxidative stress up‐regulates presenilin 1 in lipid rafts in neuronal cells
Authors: Oda Akiko , Tamaoka Akira , Araki Wataru 
Published: November 2009

AbstractOxidative stress is associated with β‐amyloid peptide (Aβ) accumulation in the brains of Alzheimer's disease patients. Aβ is generated upon the sequential proteolytic cleavage of transmembrane amyloid precursor protein (APP) by two membrane‐bound proteases, β‐secretase (BACE1) and the γ‐secretase complex comprising presenilin 1 (PS1), nicastrin, APH‐1 and PEN‐2. Recent evidence suggests that significant amounts of BACE1 and γ‐secretase components localize in the cholesterol‐rich region of membranes known as lipid rafts, where Aβ production occurs preferentially. In this study, we investigated the effects of oxidative stress on the BACE1 and γ‐secretase components in lipid rafts using human neuroblastoma SH‐SY5Y cells exposed to ethacrynic acid (EA), a compound that induces cellular glutathione depletion. Following exposure of cells to EA, heme oxygenase‐1, a marker protein of oxidative stress, was strongly induced. Moreover, treatment with EA resulted in a significant increase in PS1 protein levels, but not those of nicastrin, APH‐1, PEN‐2 or BACE1, in both cell lysates and the lipid raft fraction. This increase in PS1 protein expression was prevented by co‐treatment with an antioxidant, N‐acetylcysteine (NAC). EA additionally induced a significant increase in PS1 mRNA expression, which was inhibited by NAC. Finally, EA treatment was found to promote Aβ secretion from cells expressing Swedish mutant APP. It appears that in our cell culture model, oxidative stress enhances PS1 protein levels in lipid rafts via up‐regulation of PS1 transcription, which may constitute the mechanism underlying the oxidative stress‐associated promotion of Aβ production. © 2009 Wiley‐Liss, Inc.
Protective role of COMP‐Ang1 in ischemic rat brain
Authors: Shin Hye Young , Lee Yoon Jin , Kim Hak Joo , Park Chul‐Kee , Kim Jong Hoon , Wang Kyu Chang , Kim Dong Gyu , Koh Gou Young , Paek Sun Ha 
Published: November 2009

AbstractIn cerebral ischemia, the induction of angiogenesis may represent a natural defense mechanism that enables the hypoxic brain to avoid progression into infarction. Angiopoietin‐1 (Ang1) is known to produce non‐leaky and stable blood vessel formation mainly by the Tie2 receptor. Therefore, we envisioned that the application of cartilage oligomeric matrix protein‐Ang1 (COMP‐Ang1), a soluble, stable, and potent form of Ang1, would promote angiogenesis and provide a protective effect following unilateral middle cerebral artery occlusion (MCAO) in rats. To this end, we employed a 2‐hour‐MCAO model, and treated rats with adenovirus encoding COMP‐Ang1 (Ade‐COMP‐Ang1) or control virus encoding β‐gal (Ade‐β‐gal). Time course magnetic resonance images (MRIs) revealed significantly reduced infarct volume in the rats treated with Ade‐COMP‐Ang1 with an improvement of post‐ischemic neurological deficits compared with rats treated with Ade‐β‐gal. Moreover, compared to the rats treated with Ade‐β‐gal, the rats treated with Ade‐COMP‐Ang1 showed an increase in blood vessels, especially in the border zone adjacent to the infarction, increased number of endogenous neuronal progenitor cells in the ischemic brain, and decreased number of TUNEL‐positive cells. Taken together, COMP‐Ang1 reduced infarct volume and consequently attenuated post‐ischemic neurological deficits through enhanced angiogenesis and increased viable cell mass of neuronal cells. © 2009 Wiley‐Liss, Inc.
Conserved fate and function of ferumoxides‐labeled neural precursor cells in vitro and in vivo
Authors: Cohen Mikhal E. , Muja Naser , Fainstein Nina , Bulte Jeff W.M. , Ben‐Hur Tamir 
Published: November 2009

AbstractRecent progress in cell therapy research for brain diseases has raised the need for non‐invasive monitoring of transplanted cells. For therapeutic application in multiple sclerosis, transplanted cells need to be tracked both spatially and temporally, in order to assess their migration and survival in the host tissue. Magnetic resonance imaging (MRI) of superparamagnetic iron oxide‐(SPIO)‐labeled cells has been widely used for high resolution monitoring of the biodistribution of cells after transplantation into the central nervous system (CNS). Here we labeled mouse glial‐committed neural precursor cells (NPCs) with the clinically approved SPIO contrast agent ferumoxides and examined their survival and differentiation in vitro, as well as their functional response to environmental signals present within the inflamed brain of experimental autoimmune encephalomyelitis (EAE) mice in vivo. We show that ferumoxides labeling does not affect NPC survival and pluripotency in vitro. Following intracerebroventricular (ICV) transplantation in EAE mice, ferumoxides‐labeled NPCs responded to inflammatory cues in a similar fashion as unlabeled cells. Ferumoxides‐labeled NPCs migrated over comparable distances in white matter tracts and differentiated equally into the glial lineages. Furthermore, ferumoxides‐labeled NPCs inhibited lymph node cell proliferation in vitro, similarly to non‐labeled cells, suggesting a preserved immunomodulatory function. These results demonstrate that ferumoxides‐based MRI cell tracking is well suited for non‐invasive monitoring of NPC transplantation. © 2009 Wiley‐Liss, Inc.
Combined usage of cascade affinity fractionation and LC‐MS/MS for the proteomics of adult mouse testis
Authors: Tan Qingqiao , Dong Dexian , Ye Long , Li Rongxiu 
Published: November 2009

AbstractIn this report, the proteomics of adult mouse testis were analyzed by the combined usage of cascade affinity fractionation and LC‐MS/MS. The differences between the selected affinity ligands in size, shape, structure, and biochemical characteristics, result in each ligand exhibiting a specific affinity to some protein groups. Therefore, a cascade composition of different ligands can be applied to the fractionation of complex tissue proteins. Ultimately, the fractions collected from cascade affinity fractionation were analyzed by LC‐MS/MS, which resulted in high confidence identification of a total of 1378 non‐redundant mouse testis protein groups, over 2.6 times as many proteins as were detected in the un‐fractionated sample (526). All detected proteins were bioinformatically categorized according to their physicochemical characteristics (such as relative molecular mass, pI, grand average hydrophobicity value, and transmembrane helices), subcellular location, and function annotation. This approach highlighted the sensitivity of this method to a wide variety of protein classes. Utilizing a combination of cascade affinity fractionation and LC‐MS/MS, we have established the largest proteomic database for adult mouse testis at the present time.