bluuebunny/crossref_metadata_2025_split huggingface dataset:
Secreted expression of human lysozyme in the yeast Pichia pastoris under the direction of the signal peptide from human serum albumin
Authors: Xiong Runsong , Chen Jingjing , Chen Jinchun 
Published: May 2008

hLM (human lysozyme) has important potential application as a future safely administered human drug and food additive. To produce secreted rhLM (recombinant hLM) from the yeast Pichia pastoris, the signal peptide from HSA (human serum albumin) was employed to direct secreted expression. On the basis of the vector pPIC3.5k, an overexpression vector, pPIC3.5k‐hLM, carrying the strong promoter AOX1 (aldehyde oxidase 1), the HSA signal peptide, the enterokinase recognition motif, the lysozyme gene and other necessary genetic segments was constructed and this was followed by a series of genetic manipulations. A positive colony was picked off to test its expression pattern. The target protein, rhLM, was obtained from the supernatant and showed a gradual enrichment with the induction time course, reaching its highest level at 72 h. This pattern was identical with that shown by the secreted expression of a heterologous protein directed by Saccharomcyes cerevisiae a‐mating factor prepro‐signal peptide in P. pastoris. After a series of purification processes, including ultrafiltration with a hollow‐fibre membrane module, DEAE‐Sepharose, Sephadex G50 chromatography and enterokinase digestion, the mature protein was characterized by MALDI–TOF‐MS/MS (matrix‐assisted laser‐desorption ionization–time‐of‐flight tandem MS), N‐terminal amino acid sequencing, and Km and Kcat determination. The results confirmed that the rhLM was identical with native hLM. Moreover, the mature protein exhibited in vitro bacteriolytic activity against the Gram‐positive bacterium Micrococcus lysodeikticus and the Gram‐negative bacterium Escherichia coli. Taken together, it appeared that the HSA signal peptide was able direct secretive expression of a heterologous protein in P. pastoris.
Neuropeptide Y expression, localization and cellular transducing effects in HUVEC
Authors: Silva Antonio P. , Kaufmann Jocelyne E. , Vivancos Cécile , Fakan Stanislav , Cavadas Claudia , Shaw Phillip , Brunner Hans R. , Vischer Ulrich , Grouzmann Eric 
Published: May 2008

Background information. NPY (neuropeptide Y) may have an effect on the properties of vascular endothelial cells such as pro‐angiogenic effects and potentiation of noradrenaline‐induced vasoconstriction. In HUVEC (human umbilical‐vein endothelial cells), immunoreactive neuropeptide Y has been detected, but NPY synthesis, storage and secretion have not been studied. The aim of the present study was to establish NPY expression, storage and cellular transducing effects in HUVEC.Results. HUVEC contain 0.19 fmol of NPY/μg of protein and 0.46 fmol of pro‐NPY/μg of protein, as measured by ELISA. RT (reverse transcriptase)—PCR confirmed the expression of NPY in HUVEC. Immunofluorescence revealed the presence of NPY in small punctate structures, with a fluorescence pattern different from that observed for von Willebrand factor, indicating distinct storage compartments. Double labelling for NPY and Rab3A demonstrated similar granular patterns, with at least partial co‐localization. Electron microscopy showed NPY immunoreactivity in vesicle‐like cytoplasmic structures, of a fine fibrillar texture, as well as in mitochondria and in the nucleus. A similar general distribution pattern was also obtained for Rab3A. Y1 and Y2 receptors were expressed in HUVEC as assessed by RT—PCR, and they were functional since NPY induced a 42 nM intracellular calcium increase within 100 s, representing 22% of the histamine‐induced response. In contrast with histamine, NPY did not induce acute von Willebrand factor secretion.Conclusions. HUVEC produce, store and respond to NPY, suggesting an autocrine regulatory role for NPY in the endothelium.
Heat‐shock response down‐regulates interleukin‐18 expression in murine peritoneal macrophages
Authors: Wang Yun , Chang Zongliang 
Published: May 2008

Background information. The heat‐shock response is a self‐defence mechanism that protects cells and organisms from a wide range of harmful stresses. Recent studies revealed that it involved the regulation of cytokine expression. Interleukin‐18 (IL‐18) is an important cytokine in mediating immune response.Results. We studied interferon‐γ (IFN‐γ)‐induced IL‐18 expression in heat‐shock‐treated murine peritoneal macrophages. Our results showed that the heat‐shock response significantly inhibited the expression of IFN‐γ‐induced pro‐inflammatory cytokine IL‐18. Interferon consensus sequence binding protein (ICSBP) is a transcription factor that binds to the promoter of IL‐18 and regulates the transcription of IL‐18. Further research on the down‐regulation mechanism showed that the DNA‐binding activity of ICSBP was greatly reduced by the heat shock response.Conclusions. These results suggest that the inhibitory effect of heat‐shock response on IL‐18 production in IFN‐γ‐stimulated macrophages is related to the suppression of the binding activity of ICSBP.
Mercury chloride decreases the water permeability of aquaporin‐4‐reconstituted proteoliposomes
Authors: Yukutake Yoshinori , Tsuji Shoji , Hirano Yoshinori , Adachi Takeshi , Takahashi Toshiyuki , Fujihara Kazuo , Agre Peter , Yasui Masato , Suematsu Makoto 
Published: May 2008

Background information. Mercurials inhibit AQPs (aquaporins), and site‐directed mutagenesis has identified Cys189 as a site of the mercurial inhibition of AQP1. On the other hand, AQP4 has been considered to be a mercury‐insensitive water channel because it does not have the reactive cysteine residue corresponding to Cys189 of AQP1. Indeed, the osmotic water permeability (Pf) of AQP4 expressed in various types of cells, including Xenopus oocytes, is not inhibited by HgCl2. To examine the direct effects of mercurials on AQP4 in a proteoliposome reconstitution system, His‐tagged rAPR4 (rat AQP4) M23 was expressed in Saccharomyces cerevisiae, purified with an Ni2+‐nitrilotriacetate affinity column, and reconstituted into liposomes with the dilution method.Results. The water permeability of AQP4 proteoliposomes with or without HgCl2 was measured with a stopped‐flow apparatus. Surprisingly, the Pf of AQP4 proteoliposomes was significantly decreased by 5 μM HgCl2 within 30 s, and this effect was completely reversed by 2‐mercaptoethanol. The dose‐ and time‐dependent inhibitory effects of Hg2+ suggest that the sensitivity to mercury of AQP4 is different from that of AQP1. Site‐directed mutagenesis of six cysteine residues of AQP4 demonstrated that Cys178, which is located at loop D facing the intracellular side, is a target responding to Hg2+. We confirmed that AQP4 is reconstituted into liposome in a bidirectional orientation.Conclusions. Our results suggest that mercury inhibits the Pf of AQP4 by mechanisms different from those for AQP1 and that AQP4 may be gated by modification of a cysteine residue in cytoplasmic loop D.
Both ERK and JNK are required for enhancement of MD‐2 gene expression during differentiation of HL‐60 cells
Authors: Li Changlin , Yu Yongshen , Wang Yun , Liu Lei , Zhang Min , Sugano Sumio , Wang Zhengguo , Chang Zongliang 
Published: May 2008

Background information. MD‐2 is associated with the extracellular domain of TLR4 (Toll‐like receptor 4) and augments TLR4‐dependent LPS (lipopolysaccharide) responses in vitro. Our previous investigation found that PMA‐induced HL‐60 cell differentiation to macrophages is associated largely with TLR2 and CD14 and, to a much lesser extent, with TLR4.Results. We studied the MD‐2 expression during differentiation of HL‐60 cells induced by PMA. The results showed that PMA, but not VitD3 (1α,25‐dihydroxy‐vitamin D3), strongly induces MD‐2 gene expression by HL‐60 cells in a time‐ and dose‐dependent manner. Treatment with an MEK [MAPK (mitogen‐activated protein kinase)/ERK (extracellular‐signal‐regulated kinase) kinase] inhibitor (PD98059) and a JNK (c‐Jun N‐terminal kinase) inhibitor (SP600125) suppresses PMA‐induced MD‐2 gene expression, whereas impairment of p38 function by treatment with the inhibitor SB203580 has no effect on MD‐2 mRNA. In order to reveal the possible molecular mechanism for such a regulation of MD‐2 gene expression, we cloned and analysed the putative MD‐2 gene promoter. Transient transfection of different deletion mutants demonstrated that the region −185/−171 (5′‐TCCTTTACAGGAAGT‐3′) of the MD‐2 gene promoter is closely related to gene transcription in response to PMA. Additionally, the transcription factor Elk‐1 has been found to bind this specific motif.Conclusions. These results suggest that ERK and JNK pathways are involved in PMA‐mediated MD‐2 gene expression during HL‐60 cell differentiation, and the activation of the MEK/possible ERK/Elk signal pathway is the mechanism responsible for PMA‐induced MD‐2 gene expression in differentiated HL‐60 cells.
S1‐1 nuclear domains: characterization and dynamics as a function of transcriptional activity
Authors: Inoue Akira , Tsugawa Katsuji , Tokunaga Kazuaki , Takahashi Kenichi P. , Uni Shigehiko , Kimura Masatsugu , Nishio Koji , Yamamoto Naoki , Honda Ken‐ichi , Watanabe Takanori , Yamane Hideo , Tani Tokio 
Published: May 2008

Background information. The RNA‐binding protein S1‐1, also called RBM10 (RNA‐binding motif 10), is a paralogue of putative tumour suppressor RBM5 and has been correlated with cancer proliferation and apoptosis. In the present study, we have investigated the cell biology of S1‐1.Results. In the extranucleolar nucleoplasm, S1‐1 occurred in hundreds of punctate and irregular domains. Some 10–40 of these domains were larger than 0.5 μm and prominent for S1‐1 immunostaining. These domains (S1‐1 nuclear bodies) were commonly present in tissue cells and in cultured cells. When cellular transcription was globally reduced by heat shock, serum starvation, culture at high cell densities or inhibition with RNA polymerase II inhibitors, small S1‐1 domains (S1‐1 granules), with weak immunostaining signals, reduced in number, whereas S1‐1 nuclear bodies became prominent and increased in size. These altered S1‐1 domains were returned to initial states when the cells were placed under normal conditions. Similar to paraspeckles, S1‐1 nuclear bodies occurred closely adjacent to nuclear speckles or IGCs (interchromatin granule clusters), as determined by immunoelectron microscopy. However, the S1‐1 nuclear bodies did not correspond to paraspeckles or IGAZs (interchromatin‐granule‐associated zones), but coincided with TIDRs (transcription‐inactivation‐dependent RNA domains), which we had characterized previously at the RNA level. The enlarged S1‐1 nuclear bodies/TIDRs accumulated the S1‐1 protein and microinjected primary and spliced mRNAs, presumably for later elevation of gene expression. In addition, electron microscopy revealed that S1‐1 was also present on perichromatin fibrils, suggesting the structure of S1‐1 granules seen at higher resolution.Conclusions. S1‐1 constitutes hundreds of nuclear domains, which dynamically change their structures in a reversible manner. Upon globally reducing RNA polymerase II transcription, S1‐1 nuclear bodies enlarge and decrease in number. They are novel domains different from paraspeckles or IGAZs, despite their similar occurrence adjacent to nuclear speckles. We discuss S1‐1 granules in terms of their association with gene expression. In addition, this is the first report of a TIDR‐localized protein.
Effect of translocator protein (18 kDa)‐ligand binding on neurotransmitter‐induced salivary secretion in rat submandibular glands
Authors: Ostuni Mariano A. , Tumilasci Omar R. , Péranzi Gabriel , Cardoso Estela M. L. , Contreras Liliana N. , Arregger Alejandro L. , Papadopoulos Vassilios , Lacapere Jean‐Jacques 
Published: May 2008

Background information. TSPO (translocator protein), previously known as PBR (peripheral‐type benzodiazepine receptor), is a ubiquitous 18 kDa transmembrane protein that participates in diverse cell functions. High‐affinity TSPO ligands are best known for their ability to stimulate cholesterol transport in organs synthesizing steroids and bile salts, although they modulate other physiological functions, including cell proliferation, apoptosis and calcium‐dependent transepithelial ion secretion. In present study, we investigated the localization and function of TSPO in salivary glands.Results. Immunohistochemical analysis of TSPO in rat salivary glands revealed that TSPO and its endogenous ligand, DBI (diazepam‐binding inhibitor), were present in duct and mucous acinar cells. TSPO was localized to the mitochondria of these cells, whereas DBI was cytosolic. As expected, mitochondrial membrane preparations, which were enriched in TSPO, exhibited a high affinity for the TSPO drug ligand,3H‐labelled PK 11195, as shown byBmaxandKdvalues of 10.0±0.5 pmol/mg and 4.0±1.0 nM respectively. Intravenous perfusion of PK 11195 increased the salivary flow rate that was induced by muscarinic and α‐adrenergic agonists, whereas it had no effect when administered alone. Addition of PK 11195 also increased the K+, Na+, Cl−and protein content of saliva, indicating that this ligand modulated secretion by acini and duct cells.Conclusions. High‐affinity ligand binding to mitochondrial TSPO modulates neurotransmitter‐induced salivary secretion by duct and mucous acinar cells of rat submandibular glands.
TGFβ1 regulates endothelial cell spreading and hypertrophy through a Rac—p38‐mediated pathway
Authors: Varon Christine , Rottiers Patricia , Ezan Jerome , Reuzeau Edith , Basoni Caroline , Kramer IJsbrand , Génot Elisabeth 
Published: May 2008

Background information. TGFβ (transforming growth factor β) is a multifunctional cytokine and a potent regulator of cell growth, migration and differentiation in many cell types. In the vascular system, TGFβ plays crucial roles in vascular remodelling, but the signalling pathways involved remain poorly characterized.Results. Using the model of porcine aortic endothelial cells, we demonstrated that TGFβ stimulates cellular spreading when cells are on collagen I. TGFβ‐stimulated Rac1–GTP accumulation, which was associated with increased MAPK (mitogen‐activated protein kinase) p38 phosphorylation. Furthermore, ectopic expression of a dominant‐negative Rac mutant, or treatment of the cells with the p38 pharmacological inhibitor SB203580, abrogated TGFβ‐induced cell spreading. Our results demonstrate for the first time that prolonged exposure to TGFβ stimulates endothelial cell hypertrophy and flattening. Collectively, these data indicate that TGFβ‐induced cell spreading and increase in cell surface areas occurs via a Rac—p38‐dependent pathway.Conclusions. The Rac—p38 pathway may have conceptual implications in pathophysiological endothelial cell responses to TGFβ, such as wound healing or development of atherosclerotic lesions.
Donor insemination and parenting: Concerns and strategies of lesbian couples. A review of international studies
Authors: WERNER CAROLINE , WESTERSTÅHL ANNA 
Published: May 2008

AbstractThe concerns and strategies of lesbian couples at the time of donor insemination, throughout pregnancy and during early parenthood differ in several ways from those of heterosexual couples. For the majority of the lesbian couples insemination was the first option to conceive. They reported having strategies for their children growing up in a heteronormative society, and the parents shared time with and responsibility for the children equally. Issues considered most important by the couple were an open and non‐judgmental attitude from caregivers without heteronormative assumptions, knowledge about lesbian women's situation and acknowledgement of the co‐mother.
Perfectionism, achievement motives, and attribution of success and failure in female soccer players
Authors: Stoeber Joachim , Becker Claudia 
Published: May 2008

While some researchers have identified adaptive perfectionism as a key characteristic to achieving elite performance in sport, others see perfectionism as a maladaptive characteristic that undermines, rather than helps, athletic performance. Arguing that perfectionism in sport contains both adaptive and maladaptive facets, the present article presents a study of N = 74 female soccer players investigating how two facets of perfectionism—perfectionistic strivings and negative reactions to imperfection (Stoeber, Otto, Pescheck, Becker, & Stoll, 2007)—are related to achievement motives and attributions of success and failure. Results show that striving for perfection was related to hope of success and self‐serving attributions (internal attribution of success). Moreover, once overlap between the two facets of perfectionism was controlled for, striving for perfection was inversely related to fear of failure and self‐depreciating attributions (internal attribution of failure). In contrast, negative reactions to imperfection were positively related to fear of failure and self‐depreciating attributions (external attribution of success) and inversely related to self‐serving attributions (internal attribution of success and external attribution of failure). It is concluded that striving for perfection in sport is associated with an adaptive pattern of positive motivational orientations and self‐serving attributions of success and failure, which may help athletic performance. In contrast, negative reactions to imperfection are associated with a maladaptive pattern of negative motivational orientations and self‐depreciating attributions, which is likely to undermine athletic performance. Consequently, perfectionism in sport may be adaptive in those athletes who strive for perfection, but can control their negative reactions when performance is less than perfect.
Cultural variation in the motivational standards of self‐enhancement and self‐criticism among bicultural Asian American and Anglo American students
Authors: Zusho Akane 
Published: May 2008

Recent work on biculturalism has made theoretical and methodological inroads into our understanding of the relation of cultural processes with psychological functioning. Through the use of cultural priming methodologies, investigators have demonstrated that biculturals, or individuals who have experienced and identify with more than one culture, can switch between various “cultural frames of reference” in response to corresponding social cues (Hong, Morris, Chiu, & Benet‐Martinez, 2000). Drawing on this work on the cognitive implications of biculturalism, the purpose of the present study was to examine the assumption that independent and interdependent self‐construals are associated with the motivational standards of self‐enhancement and self‐criticism, respectively. More specifically, the effects of differential primes of self on ratings of self‐enhancement were investigated in a sample of bicultural Asian American (N = 42) and Anglo American (N = 60) college students; overall, more similarities than differences were noted between the two groups. It was hypothesized that Anglo American students would display marked tendencies toward self‐enhancement. However, this hypothesis was not supported. Nevertheless, consistent prime effects were observed for a selected number of ratings related to academic virtues, with those who received an independent‐self prime often exhibiting greater self‐enhancing tendencies than those who received an interdependent‐self prime. For example, participants in the independent‐self condition reported on average significantly higher ratings for self‐discipline and initiative, as well as the degree to which they perceived themselves to be hard working. Implications for the work on self‐representations, motivation, and acculturation are discussed.
Increased expression of vascular endothelial growth factor in hepatocellular carcinoma after transcatheter arterial chemoembolization
Authors: Wang B. , Xu H. , Gao Z. Q. , Ning H. F. , Sun Y. Q. , Cao G. W. 
Published: May 2008

Background: Changes in the biological behavior of residual viable hepatocellular carcinoma (HCC) tissue after transcatheter arterial chemoembolization (TACE) remain unclear. Several studies have reported that TACE inhibits tumor angiogenesis and induces tumor cell apoptosis, while other studies have found that TACE stimulates tumor angiogenesis and thus increases the proliferative activity of the tumor cells to some degree. Purpose: To investigate the intratumoral microvessel density (MVD) and vascular endothelial growth factor (VEGF) expression in residual surviving cancerous tissue after TACE in HCC. Material and Methods: Tumor specimens from 63 histopathologically diagnosed patients were studied: 42 comprising the control group (treated by surgery alone) and 21 comprising the TACE group (those treated by TACE 1–2 times prior to surgical resection). The number of VEGF-positive cells, MVD, and microvessel diameter were measured. Results: The MVD was 51.69±18.17 and 58.57±15.75 in the control and TACE groups, respectively. There was no significant difference between the two groups ( t=1.48, P>0.05). The microvessel diameter was 17.62±10.54 µm and 15.79±7.65 µm in the control and TACE groups, respectively, indicating no significant difference between the two groups ( t=0.71, P>0.05). The number of VEGF-positive cells in the TACE group, i.e., 243.66±88.88, was higher than that in the control group, i.e., 138.26±65.24 ( t=5.34, P<0.01). TACE increased VEGF expression in the residual surviving HCC tissues, and there was a positive correlation between VEGF expression and MVD ( r=0.4936, t=4.4329, P<0.05) in the HCC tissue. Conclusion: The study indicates that the residual surviving cancerous tissue in HCC after TACE has a rich vascularity. TACE increases VEGF expression in the residual surviving cancerous tissue.
Focal pancreatitis mimicking pancreatic mass: magnetic resonance imaging (mri)/magnetic resonance cholangiopancreatography (mrcp) findings including diffusion-weighted mri
Authors: Momtahen A. J. , Balci N. C. , Alkaade S. , Akduman E. I. , Burton F. R. 
Published: May 2008

Background: Focal pancreatitis (FP) is a confined inflammation that mimics a pancreatic mass. Its imaging diagnosis is important to avoid unnecessary procedures. Purpose: To describe the spectrum of magnetic resonance imaging (MRI)/magnetic resonance cholangiopancreatography (MRCP) and diffusion-weighted MRI (DWI) findings of focal pancreatitis mimicking pancreatic masses. Material and Methods: Findings of MRI/MRCP including DWI with a b value of 0 and 600 s/mm2 in 14 patients with pancreatic masses on MRI were retrospectively reviewed and compared to normal pancreas in 14 patients as a control group. Results: FP revealed hypointense signal intensity (SI) (3/14), hypo- to isointense SI (7/14), or isointense SI (4/14) on T1-weighted images, and hypointense SI (1/14), isointense SI (5/14), iso- to hyperintense SI (7/14), or hyperintense SI (1/14) on T2-weighted images compared to remaining pancreas (RP). MRCP images revealed dilatation of the common bile duct (CBD) and main pancreatic duct (MPD) (5/14), dilatation of the MPD only (3/14), dilatation of the CBD only (3/14), and normal MPD and CBD (3/14). Both FP and RP revealed three types of time–signal intensity curves: 1) rapid rise to a peak, with a rapid decline (FP=2, RP=4), 2) slow rise to a peak, followed by a slow decline (FP=5, RP=4), and 3) slower rise to a peak, with a slow decline or plateau (FP=7, RP=6). Mean apparent diffusion coefficient (ADC) values for FP and RP were 2.09±0.18 and 2.03±0.2×10−3 mm2/s, respectively. ADC values of FP and RP revealed no significant difference. Conclusion: The spectrum of imaging findings of focal pancreatitis on MRI/MRCP including DWI was described. Findings of FP were not distinctive as compared to the remaining pancreas.