bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cell-cycle-regulated phosphorylation of cAMP response element-binding protein: identification of novel phosphorylation sites
Authors: SAEKI Kumiko , YUO Akira , TAKAKU Fumimaro 
Published: August 2015

We report that the cAMP response element binding protein (CREB) undergoes cell-cycle-regulated phosphorylation. In human amnion FL cells, CREB was expressed as two forms with different molecular masses, 45 and 45.5 kDa. Although asynchronous cells contained predominantly the 45 kDa forms, this form shifted to 45.5 kDa when the cells were synchronized with the early S-phase. Furthermore the expression of the 45.5 kDa band was increased when cells were treated with okadaic acid, confirming that the 45.5 kDa band was a phosphorylated form of the 45 kDa band. Mutation analysis indicated that neither Ser133, the target of cAMP-dependent protein kinase and calcium calmodulin kinase, nor Ser129, the target of glycogen synthetase kinase 3, was responsible for the expression of the 45.5 kDa band, but that Ser108, Ser111 and Ser114, located in a region matching the consensus sequence for the casein kinase II target, were required. A mutant in which Ser111 and Ser114 were each replaced by a glutamic residue, mimicking a phosphorylated state, had a higher activation potential in cAMP response element-mediated transcription. These results strongly suggest that the casein kinase II target region is involved in cell cycle-regulated phosphorylation of the CREB protein and also in transcriptional enhancement.
Regulation of the gene for human dipeptidyl peptidase IV by hepatocyte nuclear factor 1α
Authors: ERICKSON Roger H. , GUM James R. , LOTTERMAN Craig D. , HICKS James W. , LAI Roy S. , KIM Young S. 
Published: August 2015

Hepatocyte nuclear factor 1 was identified as the transcription factor binding to a 20 bp (-150 to -131) region of the gene for human dipeptidyl peptidase IV, which has been shown to be important for the expression of dipeptidyl peptidase IV in the human intestinal and hepatic epithelial cell lines Caco-2 and HepG2. Functional analysis of the hepatocyte nuclear factor 1 site was performed with two minimal dipeptidyl peptidase IV promoter constructs (-250 to -41, and -150 to -41) with and without a 3 bp mutation in the hepatocyte nuclear factor 1 sequence, and used in transient transfection experiments with Caco-2 cells. The results show that the mutated constructs were able to drive transcription at only 5–10% of the activity of the non-mutated controls. Co-transfection of 3T3 cells with hepatocyte nuclear factor 1 (α or β) and dipeptidyl peptidase IV promoter constructs (-250 to -41 or -150 to -41) resulted in a 2.5–6-fold increase in transcription over controls with hepatocyte nuclear factor 1α but not with hepatocyte nuclear factor 1β. The results of this study show that hepatocyte nuclear factor 1 binds to the -150 to -131 region of the human dipeptidyl peptidase IV promoter and is necessary for transcriptional activation of the gene for dipeptidyl peptidase IV.
Agonist-induced desensitization and phosphorylation of m1-muscarinic receptors
Authors: WAUGH Mark G. , CHALLISS R. A. John , BERSTEIN Gabriel , NAHORSKI Stefan R. , TOBIN Andrew B. 
Published: August 2015

Pre-stimulation of Chinese hamster ovary (CHO) cells expressing the human m1-muscarinic receptor (CHO-m1 cells) with a maximally effective concentration of the muscarinic agonist methacholine resulted in desensitization of Ins(1,4,5)P3 accumulation, apparent as a ∼ 4-fold shift in the agonist dose–response curve. Agonist-induced desensitization was rapid (detectable by 10 s) and concentration dependent (EC50 = 8.2±2.2 µM) and resulted in a complete loss of receptor reserve for the agonist-stimulated Ins(1,4,5)P3 response. An investigation of the possible mechanisms involved in m1-muscarinic receptor desensitization indicated that agonist-induced receptor internalization, PtdIns-(4,5)P2 depletion or an increased rate of Ins(1,4,5)P3 metabolism were not involved. m1-Muscarinic receptors did, however, undergo rapid agonist-induced phosphorylation with a time course that was consistent with an involvement in receptor desensitization. Characterization studies indicated that the receptor-specific kinase involved was distinct from protein kinase C and other second-messenger-dependent protein kinases. Since previous studies have suggested that the m3-muscarinic receptor subtype undergoes agonist-dependent phosphorylation via casein kinase 1α (CK1α) [Tobin, Totty, Sterlin and Nahorski (1997) J. Biol. Chem. 272, 20844–20849], we examined the ability of m1-muscarinic receptors to be phosphorylated by this kinase. In reconstitution experiments, CK1α was able to phosphorylate purified, soluble m1-muscarinic receptors in an agonist-dependent manner.
Differential regulation of vascular endothelial growth factor and its receptor fms-like-tyrosine kinase is mediated by nitric oxide in rat renal mesangial cells
Authors: FRANK Stefan , STALLMEYER Birgit , KÄMPFER Heiko , SCHAFFNER Christian , PFEILSCHIFTER Josef 
Published: August 2015

Under conditions associated with local and systemic inflammation, mesangial cells and invading immune cells are likely to be responsible for the release of large amounts of nitric oxide (NO) in the glomerulus. To further define the mechanisms of NO action in the glomerulus, we attempted to identify genes which are regulated by NO in rat glomerular mesangial cells. We identified vascular endothelial growth factor (VEGF) and its receptor fms-like tyrosine kinase (FLT-1) to be under the regulatory control of exogenously applied NO in these cells. Using S-nitroso-glutathione (GSNO) as an NO-donating agent, VEGF expression was strongly induced, whereas expression of its FLT-1 receptor simultaneously decreased. Expressional regulation of VEGF and FLT-1 mRNA was transient and occurred rapidly within 1–3 h after GSNO treatment. Expression of a second VEGF-specific receptor, fetal liver kinase-1 (FLK-1/KDR), could not be detected. The inflammatory cytokine interleukin-1β mediated a moderate increase in VEGF expression after 24 h and had no influence on FLT-1 expression. In contrast, platelet-derived growth factor–BB and basic fibroblast growth factor had no effect on VEGF expression, but strongly induced FLT-1 mRNA levels. Obviously, there is a differential regulation of VEGF and its receptor FLT-1 by NO, cytokines and growth factors in rat mesangial cells.
Thyroid hormone promotes the phosphorylation of STAT3 and potentiates the action of epidermal growth factor in cultured cells
Authors: LIN Hung-Yun , SHIH Ai , DAVIS Faith B. , DAVIS Paul J. 
Published: August 2015

We have examined the effects of l-thyroxine (T4) on the activation of signal transducer and activator of transcription 3 (STAT3) and on the STAT3-dependent induction of c-Fos expression by epidermal growth factor (EGF). T4, at a physiological concentration of 100 nM, caused tyrosine phosphorylation and nuclear translocation (i.e. activation) of STAT3 in HeLa cells in as little as 10–20 min. Activation by T4 of STAT3 was maximal at 30 min (15±4-fold enhancement; mean±S.E.M.) in 18 experiments. This effect was reproduced by T4–agarose (100 nM) and blocked by CGP 41251, genistein, PD 98059 and geldanamycin, inhibitors of protein kinase C (PKC), protein tyrosine kinase (PTK), mitogen-activated protein kinase (MAPK) kinase and Raf-1 respectively. Tyrosine-phosphorylated MAPK also appeared in nuclear fractions within 10 min of treatment with T4. In the nuclear fraction of T4-treated cells, MAPK immunoprecipitate also contained STAT3. The actions of T4 were similar in HeLa and CV-1 cells, which lack thyroid hormone receptor (TR), and in TR-replete skin fibroblasts (BG-9). T4 also potentiated the EGF-induced nuclear translocation of activated STAT1α and STAT3 and enhanced the EGF-stimulated expression of c-Fos. Hormone potentiation of EGF-induced signal transduction and c-Fos expression was inhibited by CGP 41251, geldanamycin and PD 98059. Therefore the non-genomically induced activation by T4 of STAT3, and the potentiation of EGF by T4, require activities of PKC, PTK and an intact MAPK pathway.
Lipoprotein-associated phospholipase A2, platelet-activating factor acetylhydrolase, generates two bioactive products during the oxidation of low-density lipoprotein: use of a novel inhibitor
Authors: MACPHEE Colin H. , MOORES Kitty E. , BOYD Helen F. , DHANAK Dash , IFE Robert J. , LEACH Colin A. , LEAKE David S. , MILLINER Kevin J. , PATTERSON Rebecca A. , SUCKLING Keith E. , TEW David G. , HICKEY Deirdre M. B. 
Published: August 2015

A novel and potent azetidinone inhibitor of the lipoprotein-associated phospholipase A2 (Lp-PLA2), i.e. platelet-activating factor acetylhydrolase, is described for the first time. This inhibitor, SB-222657 (Ki = 40±3 nM, kobs/[I] = 6.6×105 M-1·s-1), is inactive against paraoxonase, is a poor inhibitor of lecithin:cholesterol acyltransferase and has been used to investigate the role of Lp-PLA2 in the oxidative modification of lipoproteins. Although pretreatment with SB-222657 did not affect the kinetics of low-density lipoprotein (LDL) oxidation by Cu2+ or an azo free-radical generator as determined by assay of lipid hydroperoxides (LOOHs), conjugated dienes and thiobarbituric acid-reacting substances, in both cases it inhibited the elevation in lysophosphatidylcholine content. Moreover, the significantly increased monocyte chemoattractant activity found in a non-esterified fatty acid fraction from LDL oxidized by Cu2+ was also prevented by pretreatment with SB-222657, with an IC50 value of 5.0±0.4 nM. The less potent diastereoisomer of SB-222657, SB-223777 (Ki = 6.3±0.5 µM, kobs/[I] = 1.6×104 M-1·s-1), was found to be significantly less active in both assays. Thus, in addition to generating lysophosphatidylcholine, a known biologically active lipid, these results demonstrate that Lp-PLA2 is capable of generating oxidized non-esterified fatty acid moieties that are also bioactive. These findings are consistent with our proposal that Lp-PLA2 has a predominantly pro-inflammatory role in atherogenesis. Finally, similar studies have demonstrated that a different situation exists during the oxidation of high-density lipoprotein, with enzyme(s) other than Lp-PLA2 apparently being responsible for generating lysophosphatidylcholine.
Physical characterization of a low-charge glycoform of the MUC5B mucin comprising the gel-phase of an asthmatic respiratory mucous plug
Authors: SHEEHAN John K. , HOWARD Marjorie , RICHARDSON Paul S. , LONGWILL Toby , THORNTON David J. 
Published: August 2015

We have previously noted that sequential extraction of an asthmatic mucous exudate with 6 M guanidinium chloride yielded a fraction of the mucins that were most resistant to solubilization and of high Mr [Sheehan, Richardson, Fung, Howard and Thornton (1995) Am. J. Respir. Cell Mol. Biol. 13, 748–756]. Here we show that this mucin fraction is dominated (at least 96% of the total) by the low-charge glycoform of the MUC5B gene product. Seen in the electron microscope the mucins appeared mainly as compact ‘island ’ structures composed of linear threads often emanating from globular ‘nodes ’ rather than the discrete linear threads more typical of mucins that we have previously described. The effect of reducing agents was as expected for other gel-forming mucins, i.e. reduced subunits or monomers of Mr 3×106) were produced within 15 min of treatment. Kinetic experiments on the cleavage of the intact mucins with the proteinase trypsin indicated two clear regimes of fragmentation. An initial rapid cleavage generated mucins ranging from Mr = 4×106 to 30×106 that in the electron microscope appeared as polydisperse threads (500–3000 nm in length), similar to normal and other respiratory mucins that we have previously characterized. A subsequent slower fragmentation over many hours yielded a major fragment of Mr 3×106 and length 200–600 nm, very similar in size and Mr to the subunits obtained by reduction. The results suggest that the MUC5B mucin is assembled, first into polydisperse linear threads, which are then linked together via a protein-mediated process. This might involve part of the mucin polypeptide or an as yet unidentified protein(s). The high proteinase susceptibility of the linkage suggests that it might be a point of control for mucin size and thus mucus rheology.
Regulation of UDP-N-acetylglucosamine:dolichyl-phosphate N-acetylglucosamine-1-phosphate transferase by retinoic acid in P19 cells
Authors: MEISSNER Joachim D. , NAUMANN Andreas , MUELLER Walter H. , SCHEIBE Renate J. 
Published: August 2015

UDP-N-acetylglucosamine:dolichyl-phosphate N-acetylglucosamine-1-phosphate transferase (GPT) is the first enzyme in the dolichol pathway of protein N-glycosylation, and is implicated in the developmental programmes of a variety of eukaryotes. In the present study we describe the effects of all-trans-retinoic acid (RA) on the levels of GPT protein and enzymic activity, and on the transcription rate of the GPT gene, in mouse P19 teratocarcinoma cells. RA caused a dose-dependent and protein-synthesis-dependent induction of enzyme activity. The maximum induction of GPT activity (about 3-fold) required 2 days of exposure to 1 µM RA. Induced GPT activity also resulted in an increase in the rate of incorporation of [3H]mannose into Glc3Man9GlcNAc2. Enzymic activities paralleled GPT gene expression. The GPT gene was induced (2-fold) after 7 h of RA treatment. An approx. 3-fold increase in a 48 kDa GPT protein and approx. 4-fold increases in the levels of three GPT transcripts (1.8, 2.0 and 2.2 kb) were observed after 2 days of RA treatment. The enhanced levels of GPT protein and mRNAs began to decline 3 days after the initiation of differentiation, and GPT expression was down-regulated during cellular differentiation. GPT activity decreased about 2.8-fold to a constant level in differentiated P19 cells. The results indicate that the RA-induced enzyme activity was mainly determined by increased transcription of the GPT gene. RA-treated P19 cells were about 4-fold more resistant to tunicamycin, a fungal antibiotic which inhibits GPT, than were control cells. In addition, GPT activity in membranes from RA-treated P19 cells exhibited approx. 4-fold increased resistance to tunicamycin compared with activity in membranes from untreated control cells, demonstrating that resistance to tunicamycin is correlated with induced GPT activity. Furthermore, increased GPT activity had regulatory significance with regard to the rate of incorporation of [3H]mannose into Glc3Man9GlcNAc2-P-P-dolichol and into glycoproteins. Together, the data provide additional insights into the hormonal regulation of GPT and present evidence that the RA-mediated induction of GPT has a regulatory impact on the dolichol pathway.
Tumour necrosis factor-α regulates expression of the CCAAT-enhancer-binding proteins (C/EBPs) α and β and determines the occupation of the C/EBP site in the promoter of the insulin-responsive glucose-transporter gene in 3T3-L1 adipocytes
Authors: JAIN Renu , POLICE Shailaja , PHELPS Kelle , PEKALA Phillip H. 
Published: August 2015

We have demonstrated previously that treatment of 3T3-L1 adipocytes with tumour necrosis factor-α (TNF) results in a rapid (4 h) and significant (75–80%) reduction in the rate of transcription of the GLUT4 gene. Control of GLUT4 gene transcription has been suggested at least in part to reside with the CCAAT-enhancer-binding protein (C/EBP) family (α, β and δ isoforms) of transcription factors. Using electrophoretic mobility shift assays, we have examined the ability of TNF to alter the occupation of the C/EBP site in the GLUT4 promoter. The data suggest that in fully differentiated adipocytes the C/EBP site is a ligand for predominantly α/α homodimers; however, after exposure to TNF, a shift in occupancy of the site occurs and the ligands become α/β heterodimers and β/β homodimers. Partner selection in dimer formation appears to be controlled by selective translocation of the β-isoform from the cytosol to the nucleus after exposure of the cells to TNF.
Conserved polar residues in the transmembrane domain of the human tachykinin NK2 receptor: functional roles and structural implications
Authors: DONNELLY Dan , MAUDSLEY Stuart , GENT J. Paul , MOSER Rachel N. , HURRELL Craig R. , FINDLAY John B. C. 
Published: August 2015

We have studied the effects of agonist and antagonist binding, agonist-induced activation and agonist-induced desensitization of the human tachykinin NK2 receptor mutated at polar residues Asn-51 [in transmembrane helix 1 (TM1)], Asp-79 (TM2) and Asn-303 (TM7), which are highly conserved in the transmembrane domain in the rhodopsin family of G-protein-coupled receptors. Wild-type and mutant receptors were expressed in both COS-1 cells and Xenopus oocytes. The results show that the N51D mutation results in a receptor which, in contrast with the wild-type receptor, is desensitized by the application of a concentration of 1 µM of the partial agonist GR64349, indicating that the mutant is more sensitive to agonist activation than is the wild-type receptor. In addition, we show that, whereas the D79E mutant displayed activation properties similar to those of the wild-type receptor, the D79N and D79A mutants displayed a severely impaired ability to activate the calcium-dependent chloride current. This suggests that it is the negative charge at Asn-79, rather than the ability of this residue to hydrogen-bond, that is critical for the activity of the receptor. Interestingly, the placement of a negative charge at position 303 could compensate for the removal of the negative charge at position 79, since the double mutant D79N/N303D displayed activation properties similar to those of the wild-type receptor. This suggests that these two residues are functionally coupled, and may even be in close proximity in the three-dimensional structure of the human tachykinin NK2 receptor. A three-dimensional model of the receptor displaying this putative interaction is presented.
A Global Measure of Judicial Independence, 1948–2012
Authors: Linzer Drew A. , Staton Jeffrey K. 
Published: August 2015

AbstractWe present a new cross-national measure of de facto judicial independence, which is available for 200 countries from 1948 to 2012. To do so, we introduce a statistical measurement model for uncovering latent concepts commonly encountered in time-series, cross-sectional analyses in comparative politics and international relations. Our approach addresses unique challenges that arise in these data: temporal dependence in the observed and unobserved variables, conceptual boundedness in the latent quantity, and substantial missing data and measurement error in the observable indicators. The resulting measures match a common conceptual definition of independence with greater reliability than existing alternatives. The model is extensible to many concepts in comparative politics and international relations.
Reflecting on hospitality management education through a practice lens
Authors: Stierand Marc , Zizka Laura 
Published: August 2015

Purpose– The purpose of this paper is to reflect on hospitality management education from a “practice epistemology” and discuss how a connecting ofsavoir(theoretical knowledge or “knowing”),savoir-faire(knowing how to do tasks, i.e. task-related skills) andsavoir-être (knowing how to be, i.e. behavior) can develop into practical knowledge.Design/methodology/approach– The purpose of the paper is achieved through novel reading of the literature on practical knowledge and formativeness applied to a higher education context.Findings– The paper suggests that it is only through the creation of context that a sensation of practicing for students can be provided, which ultimately may lead to practical knowledge. Context must be actively created through situations that invite participation to explore the logic of practice. Therefore,savoirshould be treated as “organizing knowing” andsavoir-faireandsavoir-êtreas “practicing knowing” to do and to be, respectively. The termssavoir,savoir-faireandsavoir-êtrewere chosen for this paper, as they were the common reference terms used in hospitality (master-) apprenticeship systems in Europe.Originality/value– The value of the paper is a personal reflection on a practice epistemology for hospitality management education from the perspective of two academic faculty members who have been practitioners in the hospitality industry and who regularly teach hospitality executives.
Developing practice oriented undergraduate courses in a quality framework
Authors: Whitelaw Paul A. , Wrathall Jeffrey 
Published: August 2015

Purpose– The purpose of this paper is to reflect upon the stakeholder, scholarly, academic and jurisdictional influences on course development for a vocationally oriented bachelor’s degree.Design/methodology/approach– This paper takes the form of a case study.Findings– Vocationally oriented bachelor’s courses can be developed, especially when the developers focus on scholarly rigour while paying due heed to jurisdictional requirements as well as the needs of key stakeholders.Research limitations/implications– This case study is limited to a specialist vocational college in Australia.Practical implications– The paper provides a framework by which others can develop vocationally oriented bachelor’s degrees.Social implications– The study can provide impetus to the development of scholarly, and academically rigorous, yet industrially relevant vocational degrees.Originality/value– This is a relatively novel paper from a non-self accrediting higher education provider.
Response Assessment of Thermal Papers from Four Continents to Fingerprint Development by Heat
Authors: Bond John W. 
Published: August 2015

AbstractLatent fingerprint deposits on thermal paper sourced from the U.S., China, the U.K., and Australia have been visualized by heating. U.S. and Chinese sourced paper produced two distinct types of fingerprint development. In one type (type 1), the paper dye colors where the deposit is present (as previously reported) and in the other type (type 2) the ‘inverse’ of this gives paper coloring only in areas not coincident with the deposit. Both development types gave identifiable fingerprints, the majority fading within 24 h of heating. Fingerprint development from U.K. and Australian sourced paper was exclusively type 1 and resistant to fading. Temperatures for fingerprint visualization were higher for U.S. paper (64–71°C) and Chinese paper (75–95°C) than for U.K. and Australian sourced paper (43–50°C). Particularly for Chinese sourced paper, these temperatures were within a few degrees of the normal paper color temperature. A mechanism for type 2 fingerprint development is proposed.
A Test of the DSP Sexing Method on CT Images from a Modern French Sample
Authors: Mestekova Sarka , Bruzek Jaroslav , Veleminska Jana , Chaumoitre Kathia 
Published: August 2015

AbstractThe hip bone is considered to be one of the most reliable indicators in sex determination. The aim of this study was to test the reliability of the DSP method for the hip bone proposed by Murail et al. (Bull Mem Soc Anthropol Paris, 17, 2005, 167) on a sample from a present‐day population in France (52 males and 54 females). Ten linear measurements were collected from three‐dimensional models derived from computed tomography images (CTI). To quantify the proportions of correct sex determinations, a more rigorous posterior probability threshold of 0.95 was applied. Using all 10 measurements, 92.3% of males and 97.2% of females were sexed correctly. The percentage of undetermined specimens varied depending on the used combination of measurements; however, all sexes were assigned with a 100% accuracy. This study proves that DSP is an appropriate and reliable tool for sex determination, based on dimensions obtained from CTI.