bluuebunny/crossref_metadata_2025 huggingface dataset:
The association of reduced folate carrier 80G>A polymorphism to outcome in childhood acute lymphoblastic leukemia interacts with chromosome 21 copy number
Authors: Gregers Jannie , Christensen Ib Jarle , Dalhoff Kim , Lausen Birgitte , Schroeder Henrik , Rosthoej Steen , Carlsen Niels , Schmiegelow Kjeld , Peterson Curt
Published: March 2010

The reduced folate carrier (RFC) is involved in the transport of methotrexate (MTX) across the cell membrane. The RFC gene (SLC19A1) is located on chromosome 21, and we hypothesized that the RFC80 G>A polymorphism would affect outcome and toxicity in childhood leukemia and that this could interact with chromosome 21 copy number in the leukemic clone. A total of 500 children with acute lymphoblastic leukemia treated according to the common Nordic treatment protocols were included, and we found that the RFC AA variant was associated with a 50% better chance of staying in remission compared with GG or GA variants (P = .046). Increased copy numbers of chromosome 21 appear to improve outcome also in children with GA or GG variant. In a subset of 182 children receiving 608 high-dose MTX courses, we observed higher degree of bone marrow toxicity in patients with the RFC AA variant compared with GA/GG variants (platelet 73 vs 99/105 × 109/L, P = .004, hemoglobin 5.6 vs 5.9/6.0 mmol/L, P = .004) and a higher degree of liver toxicity in patients with RFC GG variant (alanine aminotransferase 167 vs 127/124 U/L, P = .05). In conclusion, the RFC 80G>A polymorphism interacts with chromosome 21 copy numbers and affects both efficacy and toxicity of MTX.
Reciprocal regulation of metabolic and signaling pathways
Authors: Barth Andreas S , Kumordzie Ami , Colantuoni Carlo , Margulies Kenneth B , Cappola Thomas P , Tomaselli Gordon F
Published: March 2010

Background

By studying genome-wide expression patterns in healthy and diseased tissues across a wide range of pathophysiological conditions, DNA microarrays have revealed unique insights into complex diseases. However, the high-dimensionality of microarray data makes interpretation of heterogeneous gene expression studies inherently difficult.

Results

Using a large-scale analysis of more than 40 microarray studies encompassing ~2400 mammalian tissue samples, we identified a common theme across heterogeneous microarray studies evident by a robust genome-wide inverse regulation of metabolic and cell signaling pathways: We found that upregulation of cell signaling pathways was invariably accompanied by downregulation of cell metabolic transcriptional activity (and vice versa). Several findings suggest that this characteristic gene expression pattern represents a new principle of mammalian transcriptional regulation. First, this coordinated transcriptional pattern occurred in a wide variety of physiological and pathophysiological conditions and was identified across all 20 human and animal tissue types examined. Second, the differences in metabolic gene expression predicted the magnitude of differences for signaling and all other pathways, i.e. tissue samples with similar expression levels of metabolic transcripts did not show any differences in gene expression for all other pathways. Third, this transcriptional pattern predicted a profound effect on the proteome, evident by differences in structure, stability and post-translational modifications of proteins belonging to signaling and metabolic pathways, respectively.

Conclusions

Our data suggest that in a wide range of physiological and pathophysiological conditions, gene expression changes exhibit a recurring pattern along a transcriptional axis, characterized by an inverse regulation of major metabolic and cell signaling pathways. Given its widespread occurrence and its predicted effects on protein structure, protein stability and post-translational modifications, we propose a new principle for transcriptional regulation in mammalian biology.
GanedenBC30™ cell wall and metabolites: anti-inflammatory and immune modulating effects in vitro
Authors: Jensen Gitte S , Benson Kathleen F , Carter Steve G , Endres John R
Published: March 2010

Background

This study was performed to evaluate anti-inflammatory and immune modulating properties of the probiotic, spore-forming bacterial strain: Bacillus coagulans: GBI-30, (PTA-6086, GanedenBC30TM). In addition, cell wall and metabolite fractions were assayed separately to address whether biological effects were due to cell wall components only, or whether secreted compounds from live bacteria had additional biological properties. The spores were heat-activated, and bacterial cultures were grown. The culture supernatant was harvested as a source of metabolites (MTB), and the bacteria were used to isolate cell wall fragments (CW). Both of these fractions were compared in a series of in vitro assays.

Results

Both MTB and CW inhibited spontaneous and oxidative stress-induced ROS formation in human PMN cells and increased the phagocytic activity of PMN cells in response to bacteria-like carboxylated fluorospheres. Both fractions supported random PMN and f-MLP-directed PMN cell migration, indicating a support of immune surveillance and antibacterial defense mechanisms. In contrast, low doses of both fractions inhibited PMN cell migration towards the inflammatory mediators IL-8 and LTB4. The anti-inflammatory activity was strongest for CW, where the PMN migration towards IL-8 was inhibited down to dilutions of 1010.

Both MTB and CW induced the expression of the CD69 activation marker on human CD3- CD56+ NK cells, and enhanced the expression of CD107a when exposed to K562 tumor cells in vitro.

The fractions directly modulated cytokine production, inducing production of the Th2 cytokines IL-4, IL-6, and IL-10, and inhibiting production of IL-2.

Both fractions further modulated mitogen-induced cytokine production in the following manner: Both fractions enhanced the PHA-induced production of IL-6 and reduced the PHA-induced production of TNF-alpha. Both fractions enhanced the PWM-induced production of TNF-alpha and IFN-gamma. In addition, MTB also enhanced both the PHA- and the PWM-induced expression of IL-10.

Conclusion

The data suggest that consumption of GanedenBC30TM may introduce both cell wall components and metabolites that modulate inflammatory processes in the gut. Both the cell wall and the supernatant possess strong immune modulating properties in vitro. The anti-inflammatory effects, combined with direct induction of IL-10, are of interest with respect to possible treatment of inflammatory bowel diseases as well as in support of a healthy immune system.
In vitro and in vivo protective efficacies of antibodies that neutralize the RNA N-glycosidase activity of Shiga toxin 2
Authors: Jeong Kwang-il , Chapman-Bonofiglio Susan , Singh Pradeep , Lee Jongo , Tzipori Saul , Sheoran Abhineet S
Published: March 2010

Backgound

Shiga toxin 2 (Stx2), one of two Stx liberated by Stx-producing Escherichia coli, is composed of an A subunit monomer and a B subunit pentamer, and is directly linked with hemolytic uremic syndrome in children. The pentameric B subunit binds to its cell surface receptor Gb3 for toxin internalization, and the A subunit follows intracellular retrograde transport to the cytosol where its RNA N-glycosidase activity (RNA-NGA) shuts down the protein synthesis, and leads to cell death. The present study investigated the ability of 19 Stx2 A subunit-specific human monoclonal antibodies (HuMAbs) to neutralize the RNA-NGA, and the association this neutralizing activity with protection of HeLa cells and mice against Stx2-induced death.

Results

The HuMAbs that were stronger inhibitors of RNA-NGA were also better at neutralizing Stx2 mediated HeLa cell death, and those that were weaker inhibitors of RNA-NGA activity were also weaker in protecting HeLa cells. These results suggest that the ability of an A subunit-specific antibody to block the RNA-NGA of the toxin is directly related to its ability to neutralize Stx2-mediated HeLa cell death. However, with the exception of the best RNA-NGA blocking antibodies 5C12 and 2F10, the efficacies of antibody neutralization of RNA-NGA of Stx2 did not correlate with their in vivo protective efficacies. The HuMAb 6C3, which neutralized RNA N-glycosidase activity of Stx2 less effectively than the HuMAbs 6D8 and 6B7, protected 100% of the mice against Stx2 challenge at 50 μg/mouse dose. In contrast, the HuMAbs 6D8 and 6B7, which neutralized RNA N-glycosidase activity of Stx2 more effectively than 6C3, protected 20% and 0% mice at that dose, respectively.

Conclusions

The neutralization efficiency of the RNA-NGA of Stx2 by A subunit-specific antibodies correlate strongly with their abilities to protect HeLa cells against Stx2-mediated toxicity but only the strongest RNA-NGA-neutralizing antibodies correlate very well with both protecting HeLa cells and mice against Stx2 challenge.
Does treatment of intestinal helminth infections influence malaria? Background and methodology of a longitudinal study of clinical, parasitological and immunological parameters in Nangapanda, Flores, Indonesia (ImmunoSPIN Study)
Authors: Wiria Aprilianto E , Prasetyani Margaretta A , Hamid Firdaus , Wammes Linda J , Lell Bertrand , Ariawan Iwan , Uh Hae Won , Wibowo Heri , Djuardi Yenny , Wahyuni Sitti , Sutanto Inge , May Linda , Luty Adrian JF , Verweij Jaco J , Sartono Erliyani , Yazdanbakhsh Maria , Supali Taniawati
Published: March 2010

Background

Given that helminth infections are thought to have strong immunomodulatory activity, the question whether helminth infections might affect responses to malaria antigens needs to be addressed. Different cross-sectional studies using diverse methodologies have reported that helminth infections might either exacerbate or reduce the severity of malaria attacks. The same discrepancies have been reported for parasitemia.

Methods/Design

To determine the effect of geohelminth infections and their treatment on malaria infection and disease outcome, as well as on immunological parameters, the area of Nangapanda on Flores Island, Indonesia, where malaria and helminth parasites are co-endemic was selected for a longitudinal study. Here a Double-blind randomized trial will be performed, incorporating repeated treatment with albendazole (400 mg) or placebo at three monthly intervals. Household characteristic data, anthropometry, the presence of intestinal helminth and Plasmodium spp infections, and the incidence of malaria episodes are recorded. In vitro cultures of whole blood, stimulated with a number of antigens, mitogens and toll like receptor ligands provide relevant immunological parameters at baseline and following 1 and 2 years of treatment rounds. The primary outcome of the study is the prevalence of Plasmodium falciparum and P. vivax infection. The secondary outcome will be incidence and severity of malaria episodes detected via both passive and active follow-up. The tertiary outcome is the inflammatory cytokine profile in response to parasite antigens. The project also facilitates the transfer of state of the art methodologies and technologies, molecular diagnosis of parasitic diseases, immunology and epidemiology from Europe to Indonesia.

Discussion

The study will provide data on the effect of helminth infections on malaria. It will also give information on anthelminthic treatment efficacy and effectiveness and could help develop evidence-based policymaking.

Trial registration

This study was approved by The Ethical Committee of Faculty of Medicine, University of Indonesia, ref:194/PT02.FK/Etik/2006 and has been filed by ethics committee of the Leiden University Medical Center. Clinical trial number:ISRCTN83830814. The study is reported in accordance with the CONSORT guidelines for cluster-randomized studies.
Promoter knock-in: a novel rational method for the fine tuning of genes
Authors: De Mey Marjan , Maertens Jo , Boogmans Sarah , Soetaert Wim K , Vandamme Erick J , Cunin Raymond , Foulquié-Moreno Maria R
Published: March 2010

Background

Metabolic engineering aims at channeling the metabolic fluxes towards a desired compound. An important strategy to achieve this is the modification of the expression level of specific genes. Several methods for the modification or the replacement of promoters have been proposed, but most of them involve time-consuming screening steps. We describe here a novel optimized method for the insertion of constitutive promoters (referred to as "promoter knock-in") whose strength can be compared with the native promoter by applying a promoter strength predictive (PSP) model.

Results

Our method was successfully applied to fine tune the ppc gene of Escherichia coli. While developing the promoter knock-in methodology, we showed the importance of conserving the natural leader region containing the ribosome binding site (RBS) of the gene of interest and of eliminating upstream regulatory elements (transcription factor binding sites). The gene expression was down regulated instead of up regulated when the natural RBS was not conserved and when the upstream regulatory elements were eliminated. Next, three different promoter knock-ins were created for the ppc gene selecting three different artificial promoters. The measured constitutive expression of the ppc gene in these knock-ins reflected the relative strength of the different promoters as predicted by the PSP model. The applicability of our PSP model and promoter knock-in methodology was further demonstrated by showing that the constitutivity and the relative levels of expression were independent of the genetic background (comparing wild-type and mutant E. coli strains). No differences were observed during scaling up from shake flask to bioreactor-scale, confirming that the obtained expression was independent of environmental conditions.

Conclusion

We are proposing a novel methodology for obtaining appropriate levels of expression of genes of interest, based on the prediction of the relative strength of selected synthetic promoters combined with an optimized promoter knock-in strategy. The obtained expression levels are independent of the genetic background and scale conditions. The method constitutes therefore a valuable addition to the genetic toolbox for the metabolic engineering of E. coli.
PPARγ gene C161T substitution alters lipid profile in Chinese patients with coronary artery disease and type 2 diabetes mellitus
Authors: Wan Jing , Xiong Shixi , Chao Shengping , Xiao Jianming , Ma Yexin , Wang Jinghua , Roy Sabita
Published: March 2010

Background

Peroxisome proliferator-activated receptor γ (PPARγ) is a ligand-activated transcription factor, which regulates gene expression of the key proteins involved in lipid metabolism, vascular inflammation, and proliferation. PPARγ may contribute to attenuating atherogenesis and postangioplasty restenosis. PPARγ C161→T substitution is associated with a reduced risk of coronary artery disease (CAD). Whether or not the gene substitution alters the risk of CAD in type 2 diabetes mellitus (T2DM) patients remains unclear.

Methods

A total of 556 unrelated subjects from a Chinese Han population, including 89 healthy subjects, 78 CAD patients, 86 T2DM patients, and 303 CAD combined with T2DM patients, were recruited to enroll in this study. PPARγC161→T gene polymorphism was determined by polymerase chain reaction and restriction fragment length polymorphisms. Plasma levels of lipoproteins, apolipoproteins, glucose, and insulin were measured by ELISA or radioimmunoassay (RIA). The coronary artery lesions were evaluated by coronary angiography.

Results

The frequency of the 161T allele in CAD, T2DM, and CAD combined with T2DM patients was similar to that observed in the healthy control group. However, in CAD combined with T2DM patients, the group with angiographically documented moderate stenoses had a higher frequency of the 161T allele in comparison to the group with severe stenoses (P < 0.05). Moreover, in CAD with T2DM patients, the triglyceride levels and apoB in CC homozygote carriers were significantly higher than those in "T" allele carriers.

Conclusions

PPARγC161→T genotypes weren't significantly associated with the risk of CAD, but were markedly correlated with severity of disease vessels in patients with CAD and T2DM. Furthermore, PPARγC161→T substitution was associated with an altered adipose, but not glucose metabolism. These results indicate that the PPARγ C161→T polymorphism may reduce the risk of severe atherogenesis by modulation of adipose metabolism, especially triglycerides and apoB, in Chinese patients with CAD and T2DM.
Direct anti-metastatic efficacy by the DNA enzyme Dz13 and downregulated MMP-2, MMP-9 and MT1-MMP in tumours
Authors: Tan Mei Lin , Choong Peter FM , Dass Crispin R
Published: March 2010

The DNA enzyme Dz13, targeted against the oncogene c-Jun, is capable of inhibiting various model tumours in mice albeit in ectopic models of neoplasia. In previous studies using orthotopic models of disease, the inhibitory effects of Dz13 on secondary growth was a direct result of growth inhibition at the primary lesion site. Thus, the direct and genuine effects on metastasis were not gauged. In this study, Dz13 was able to inhibit both locoregional and distal metastasis of tumour cells in mice, in studies where the primary tumours were unaffected due to the late and clinically-mimicking nature of treatment commencement. In addition, the effect of Dz13 against tumours has now been extended to encompass breast and prostate cancer. Dz13 upregulated the matrix metalloproteinase (MMP)-2 and MMP-9, and decreased expression of MT1-MMP (MMP-14) in cultured tumour cells. However, in sections of ectopic tumours treated with Dz13, both MMP-2 and MMP-9 were downregulated. Thus, not only is Dz13 able to inhibit tumour growth at the primary site, but also able to decrease the ability of neoplastic cells to metastasise. These findings further highlight the growing potential of Dz13 as an antineoplastic agent.
A gel-free approach in vascular smooth muscle cell proteome: perspectives for a better insight into activation
Authors: Rocchiccioli Silvia , Citti Lorenzo , Boccardi Claudia , Ucciferri Nadia , Tedeschi Lorena , Lande Caterina , Trivella Maria Giovanna , Cecchettini Antonella
Published: March 2010

Background

The use of chromatography coupled with mass spectrometry (MS) analysis is a powerful approach to identify proteins, owing to its capacity to fractionate molecules according to different chemical features. The first protein expression map of vascular smooth muscle cells (VSMC) was published in 2001 and since then other papers have been produced. The most detailed two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) map was presented by Mayr et al who identified 235 proteins, corresponding to the 154 most abundant unique proteins in mouse aortic VSMC. A chromatographic approach aimed at fractionating the VSMC proteome has never been used before.

Results

This paper describes a strategy for the study of the VSMC proteome. Our approach was based on pre-fractionation with ion exchange chromatography coupled with matrix assisted laser desorption-time of flight mass spectrometry analysis assisted by a liquid chromatography (LC-MALDI-TOF/TOF). Ion exchange chromatography resulted in a good strategy designed to simplify the complexity of the cellular extract and to identify a large number of proteins. Selectivity based on the ion-exchange chemical features was adequate if evaluated on the basis of protein pI. The LC-MALDI approach proved to be highly reproducible and sensitive since we were able to identify up to 815 proteins with a concentration dynamic range of 7 orders of magnitude.

Conclusions

In our opinion, the large number of identified proteins and the promising quantitative reproducibility made this approach a powerful method to analyze complex protein mixtures in a high throughput way and to obtain statistical data for the discovery of key factors involved in VSMC activation and to analyze a label-free differential protein expression.
Papillomavirus pseudovirions packaged with the L2 gene induce cross-neutralizing antibodies
Authors: Combelas Nicolas , Saussereau Emilie , Fleury Maxime JJ , Ribeiro Tatiana , Gaitan Julien , Duarte-Forero Diego F , Coursaget Pierre , Touzé Antoine
Published: March 2010

Background

Current vaccines against HPVs are constituted of L1 protein self-assembled into virus-like particles (VLPs) and they have been shown to protect against natural HPV16 and HPV18 infections and associated lesions. In addition, limited cross-protection has been observed against closely related types. Immunization with L2 protein in animal models has been shown to provide cross-protection against distant papillomavirus types, suggesting that the L2 protein contains cross-neutralizing epitopes. However, vaccination with L2 protein or L2 peptides does not induce high titers of anti-L2 antibodies. In order to develop a vaccine with the potential to protect against other high-risk HPV types, we have produced HPV58 pseudovirions encoding the HPV31 L2 protein and compared their capacity to induce cross-neutralizing antibodies with that of HPV L1 and HPV L1/L2 VLPs.

Methods

The titers of neutralizing antibodies against HPV16, HPV18, HPV31 and HPV58 induced in Balb/c mice were compared after immunization with L2-containing vaccines.

Results

Low titers of cross-neutralizing antibodies were detected in mice when immunized with L1/L2 VLPs, and the highest levels of cross-neutralizing antibodies were observed in mice immunized with HPV 58 L1/L2 pseudovirions encoding the HPV 31 L2 protein.

Conclusions

The results obtained indicate that high levels of cross-neutralizing antibodies are only observed after immunization with pseudovirions encoding the L2 protein. HPV pseudovirions thus represent a possible new strategy for the generation of a broad-spectrum vaccine to protect against high-risk HPVs and associated neoplasia.
The cytoprotective drug amifostine modifies both expression and activity of the pro-angiogenic factor VEGF-A
Authors: Dedieu S , Canron X , Rezvani HR , Bouchecareilh M , Mazurier F , Sinisi R , Zanda M , Moenner M , Bikfalvi A , North S
Published: March 2010

Background

Amifostine (WR-2721, delivered as Ethyol®) is a phosphorylated aminothiol compound clinically used in addition to cis-platinum to reduce the toxic side effects of therapeutic treatment on normal cells without reducing their efficacy on tumour cells. Its mechanism of action is attributed to the free radical scavenging properties of its active dephosphorylated metabolite WR-1065. However, amifostine has also been described as a potent hypoxia-mimetic compound and as a strong p53 inducer; both effects are known to potently modulate vascular endothelial growth factor (VEGF-A) expression. The angiogenic properties of this drug have not been clearly defined.

Methods

Cancer cell lines and endothelial cells were used in culture and treated with Amifostine in order to study (i) the expression of angiogenesis related genes and proteins and (ii) the effects of the drug on VEGF-A inducedin vitroangiogenesis.

Results

We demonstrated that the treatment of several human cancer cell lines with therapeutical doses of WR-1065 led to a strong induction of different VEGF-A mRNA isoforms independently of HIF-1α. VEGF-A induction by WR-1065 depends on the activation of the eIF2alpha/ATF4 pathway. This up-regulation of VEGF-A mRNA was accompanied by an increased secretion of VEGF-A proteins fully active in stimulating vascular endothelial cells (EC). Nevertheless, direct treatment of EC with amifostine impaired their ability to respond to exogenous VEGF-A, an effect that correlated to the down-regulation of VEGFR-2 expression, to the reduction in cell surface binding of VEGF-A and to the decreased phosphorylation of the downstream p42/44 kinases.

Conclusions

Taken together, our results indicate that amifostine treatment modulates tumour angiogenesis by two apparently opposite mechanisms - the increased VEGF-A expression by tumour cells and the inhibition of EC capacity to respond to VEGF-A stimulation.
Keratin 18 phosphorylation as a progression marker of chronic hepatitis B
Authors: Shi Ying , Sun Shihui , Liu Yali , Li Junfeng , Zhang Tong , Wu Hao , Chen Xinyue , Chen Dexi , Zhou Yusen
Published: March 2010

Background

The intermediate filament proteins keratins 18 (K18) and 8 (K8) polymerize to form the cytoskeletal network in the mature hepatocytes. It has been shown that the phosphorylation of K18 at two serine residues, 33 and 52, correlates with the progression of hepatitis C, but little is known of chronic hepatitis B (CHB). In this study, we examined K18 phosphorylation in relation to CHB.

Results

Site-specific phosphorylation of K18 was determined in livers of twelve healthy donors, and non-cirrhosis (n = 40) and cirrhosis (n = 21) patients. On average, progressively higher level of Ser52 phosphorylation was observed in non-cirrhotic and cirrhotic livers, while elevated Ser33 phosphorylation was detected in both livers but no significant difference. Progressive increase of Ser33 and Ser52 phosphorylation correlated with the elevation of both histological lesions and enzymatic activities of alanine aminotransferase in non-cirrhotic livers. In the hepatocytes of an inactive HBV carrier, strong signals of Ser33 phosphorylation were co-localized with viral infection, while only basal level of Ser52 phosphorylation was detected in infected cells.

Conclusion

Assuming all obtained data, our data suggest that K18 phosphorylation is a progression marker for CHB.
Paraneoplastic limbic encephalitis presenting as a neurological emergency: a case report
Authors: Xia Zongqi , Mehta Brijesh P , Ropper Allan H , Kesari Santosh
Published: March 2010

Introduction

Paraneoplastic limbic encephalitis remains a challenging clinical diagnosis with poor outcome if it is not recognized and treated early in the course of the disease.

Case Presentation

A 65-year-old Caucasian woman presented with generalized tonic-clonic seizures and increasing confusion shortly after a lung biopsy that led to the diagnosis of small-cell lung cancer. She had a complicated hospital course, and had recurrent respiratory distress due to aspiration pneumonia, and fluctuating mental status and seizures that were refractory to anti-epileptic drug treatment. Routine laboratory testing, magnetic resonance imaging of the brain, electroencephalogram, lumbar puncture, serum and cerebrospinal fluid tests for paraneoplastic antibodies, and chest computed tomography were performed on our patient. The diagnosis was paraneoplastic limbic encephalitis in the setting of small-cell lung cancer with positive N-type voltage-gated calcium channel antibody titer. Anti-epileptic drugs for seizures, chemotherapy for small-cell lung cancer, and intravenous immunoglobulin and steroids for paraneoplastic limbic encephalitis led to a resolution of her seizures and improved her mental status.

Conclusion

Early recognition of paraneoplastic limbic encephalitis and prompt intervention with immune therapies at the onset of presentation will probably translate into more favorable neurological outcomes.
Expression and aberrant promoter methylation of Wnt inhibitory factor-1 in human astrocytomas
Authors: Yang Zhuanyi , Wang Ying , Fang Jiasheng , Chen Fenghua , Liu Jinfang , Wu Jun , Wang Yanjin
Published: March 2010

Background

Wnt inhibitory factor-1(WIF-1) acts as a Wnt-antagonists and tumor suppressor, but hypermethylation of WIF-1 gene promoter and low expression activate Wnt signaling aberrantly and induce the development of various human tumors. With this work we intended to investigate the expression and promoter methylation status of WIF-1 gene in human astrocytomas.

Methods

The tissue samples consisted of 53 astrocytomas and 6 normal brain tissues. The expression levels of WIF-1 were determined by immunohistochemistry and semiquantitative RT-PCR. The results were analyzed in correlation with clinicopathological data. Methylation status of WIF-1 gene promoter was investigated using methylation specific PCR. The relationship between methylation and expression of the genes was analyzed.

Results

The average expression levels of WIF-1 protein and mRNA in astrocytomas were decreased significantly compared with normal control tissues. The protein and mRNA expression of WIF-1 gene in astrocytomas was decreased with the increase of pathological grade. Furthermore, WIF-1 promoter methylation was observed by MS-PCR in astrocytomas which showed significant reduction of WIF-1 expression. The WIF-1 promoter hypermethylation was associated with reduced expression of WIF-1 expression.

Conclusion

Our results demonstrate that the WIF-1 gene is frequently down-regulated or silenced in astrocytomas by aberrant promoter methylation. This may be an important mechanism in astrocytoma carcinogenesis.
No Trace of the Bland: An Interview with Holly Hughes
Authors: Carr C.
Published: March 2010

Holly Hughes began her career at New York's wow Cafe, that “home for wayward girls” featuring work by lesbians. There she developed her first play, The Well of Horniness (1983)—now a classic in certain circles. Hughes moved very quickly from that playful soap opera parody to experimental non-narrative pieces such as The Lady Dick (1985) and Dress Suits to Hire (1987). All of Hughes's work has an undercurrent of dark comic rage. And her subject has always been relationships between women. Despite her notoriety as a “lesbian artist,” this does not necessarily mean sexual relationships. Her characters do tend to drive each other to emotional extremes; they do put their sexuality up front. But they are basically trying to understand their own identities in a world where they don't have a place.

In 1990, Hughes was touring in World Without End, a monologue inspired by the death of her mother, when she and three other performance artists—Karen Finley, John Fleck, and Tim Miller—were defunded by then-chairman of the National Endowment for the Arts, John Frohnmayer. Although the chairman declared that the work of these artists lacked artistic merit, the NEA 4 went to court, charging that they'd been defunded for political reasons. Never interested in offering simple answers or definitions—especially for a word like “lesbian”—Hughes now found such simplifications both demanded of her and projected onto her. Becoming visible for all the wrong reasons brought daily reminders of her lesbianism as an embattled identity, but in 1992, Hughes completed a new play about racism, No Trace of the Blonde. And she continues the task of extricating herself and her work from the distorting notoriety that surrounds the censorship crisis.
Reports from the 21st Century: A Sarajevo Interview
Authors: Munk Erika
Published: March 2010

The following interview was done backstage at the Sarajevo Youth Theater, August 25, 1993, with actors from its productions of Alkestis and Waiting for Godot. Haris Pasovic—professor at Sarjevo's Performing Arts Academy, Alkestis's director, and head of the Sarajevo Theater Festival, translated with miraculous speed and stamina. The participants are Amina Begovic (Alcestis 2 in Alkestis); Izudin Bajrovic (Hercules in Alkestis, Vladimir in Waiting for Godot); Sead Bejtovic (Admetus and the second Vladimir); Admir Glamocak (the second Death in Alkestis, Lucky); Mirza Halilovic (composer and leader of the chorus for Alkestis, the boy in Godot); Jasna Huseinovic (second chorus leader); Emina Muftic (Alcestis 1); Irena Mulamuhic (Estragon and a servant in Alkestis).
Bellarmine's Revenge? On Some Recent Trends in the Roman Catholic Church Concerning the Relation of Faith and Science
Authors: De Caro Mario , Pievani Telmo
Published: March 2010

For many centuries, since the beginning of the modern age, the Roman Catholic Church held an ambivalent attitude toward science, until an enlightened position seemed to prevail at the end of the twentieth century, with two papal statements that respectively concerned Galileo's trial and Darwin's theory. In the last few years, however, within the higher circles of the Vatican, a less open-minded stance with regard to both the natural and the social sciences seems to have gained strength again.

In this article, the roots, nature, and implications of this less-tolerant trend are discussed by analyzing two key case studies. The first concerns the view of the Roman Catholic Church with regard to the Darwinian theory of evolution (which, surprisingly, seems now less accepted than it used to be in the recent past, in favor of views clearly at odds with the scientific worldview, such as purposeful evolution or even intelligent design). The second case study concerns the methodological issue of how Jesus's historical figure should be studied.
Affirming the Human in China
Authors: Davies Gloria
Published: March 2010

In engaging with Chinese perceptions of human rights, we must first consider how the human is understood in Chinese. The Confucian idea of the human as synonymous with benevolence is a morally exacting one. Across the centuries, it has found articulation in the enduring demand for educated persons to conduct themselves as moral exemplars. In this essay, the divergent ramifications of this Confucian dictum are explored via several Chinese thinkers including Huang Zongxi, Liang Qichao, Lu Xun, and Mao Zedong. Their intellectual legacies are considered in relation to present-day uses of exemplarity in the People's Republic that reflect the continued importance of Confucian benevolence as a guiding concept. The essay highlights rhetorical similarities between official discourse and the discourse of dissidents to draw attention to a common historical presumption in both to speak on behalf of the Chinese people. The argument I present is that, no matter how diverse their political goals, state representatives and dissidents share a common intellectual ancestry in the Confucian language of moral exemplarity. A large part of the discussion revolves around the Confucian notion of “cleaving to the true path” as having an affective force that tends toward the magisterial and the prescriptive. The essay explores, in this context, the dangers associated with anticipating “the true path” discursively structured around the figure of the selfless, exemplary human.
Abnormal hair follicle development and altered cell fate of follicular keratinocytes in transgenic mice expressing ΔNp63α
Authors: Romano Rose-Anne , Smalley Kirsten , Liu Song , Sinha Satrajit
Published: March 2010

The transcription factor p63 plays an essential role in epidermal morphogenesis. Animals lacking p63 fail to form many ectodermal organs, including the skin and hair follicles. Although the indispensable role of p63 in stratified epithelial skin development is well established, relatively little is known about this transcriptional regulator in directing hair follicle morphogenesis. Here, using specific antibodies, we have established the expression pattern of ΔNp63 in hair follicle development and cycling. ΔNp63 is expressed in the developing hair placode, whereas in mature hair its expression is restricted to the outer root sheath (ORS), matrix cells and to the stem cells of the hair follicle bulge. To investigate the role of ΔNp63 in hair follicle morphogenesis and cycling, we have utilized a Tet-inducible mouse model system with targeted expression of this isoform to the ORS of the hair follicle. ΔNp63 transgenic animals display dramatic defects in hair follicle development and cycling, eventually leading to severe hair loss. Strikingly, expression of ΔNp63 leads to a switch in cell fate of hair follicle keratinocytes, causing them to adopt an interfollicular epidermal (IFE) cell identity. Moreover, ΔNp63 transgenic animals exhibit a depleted hair follicle stem-cell niche, which further contributes to the overall cycling defects observed in the mutant animals. Finally, global transcriptome analysis of transgenic skin identified altered expression levels of crucial mediators of hair morphogenesis, including key members of the Wnt/β-catenin signaling pathway, which, in part, account for these effects. Our data provide evidence supporting a role for ΔNp63α in actively suppressing hair follicle differentiation and directing IFE cell lineage commitment.
The NPIY motif in the integrin β1 tail dictates the requirement for talin-1 in outside-in signaling
Authors: Nieves Bethsaida , Jones Christopher W. , Ward Rachel , Ohta Yasutaka , Reverte Carlos G. , LaFlamme Susan E.
Published: March 2010

Protein interactions with the integrin β-subunit cytoplasmic domain (β-tail) are essential for adhesion-dependent processes, including cell spreading and the connection of integrins with actin filaments at adhesion sites. Talin-1 binds to the conserved membrane-proximal NPxY motif of β-tails (NPIY in β1 integrin) promoting the inside-out activation of integrins and providing a linkage between integrins and the actin cytoskeleton. Here, we characterize the role of interactions between talin-1 and β-tail downstream of integrin activation, in the context of recombinant integrins containing either the wild type (WT) or the (YA) mutant β1A tail, with a tyrosine to alanine substitution in the NPIY motif. In addition to inhibiting integrin activation, the YA mutation suppresses cell spreading, integrin signaling, focal adhesion and stress-fiber formation, as well as microtubule assembly. Constitutive activation of the mutant integrin restores these integrin-dependent processes, bringing into question the importance of the NPIY motif downstream of integrin activation. Depletion of talin-1 using TLN1 siRNA demonstrated that talin-1 is required for cell spreading, focal adhesion and stress-fiber formation, as well as microtubule assembly, even when cells are adhered by constitutively activated WT integrins. Depletion of talin-1 does not inhibit these processes when cells are adhered by constitutively activated mutant integrins, suggesting that the binding of an inhibitory protein to the NPIY motif negatively regulates integrin function when talin-1 is depleted. We identified filamin A (FLNa) as this inhibitory protein; it binds to the β1A tail in an NPIY-dependent manner and inhibition of FLNa expression in talin-1-depleted cells restores integrin function when cells are adhered by constitutively activated WT integrins. FLNa binds FilGAP, which is a negative regulator of Rac activation. Expression of the dominant inhibitory mutant, FilGAPΔGAP, which lacks GAP activity restores spreading in cells adhered by constitutively activated integrins containing the β1A tail, but not by integrins containing the β1D tail, which is known to bind poorly to FLNa. Together, these results suggest that the binding of talin-1 to the NPIY motif is required downstream of integrin activation to promote cell spreading by preventing the inappropriate recruitment of FLNa and FilGAP to the β1A tail. Our studies emphasize the importance of understanding the mechanisms that regulate the differential binding FLNa and talin-1 to the β1 tail downstream of integrin activation in promoting integrin function.
The hinge region of the human estrogen receptor determines functional synergy between AF-1 and AF-2 in the quantitative response to estradiol and tamoxifen
Authors: Zwart Wilbert , de Leeuw Renée , Rondaij Mariska , Neefjes Jacques , Mancini Michael A. , Michalides Rob
Published: March 2010

Human estrogen receptors α and β (ERα and ERβ) greatly differ in their target genes, transcriptional potency and cofactor-binding capacity, and are differentially expressed in various tissues. In classical estrogen response element (ERE)-mediated transactivation, ERβ has a markedly reduced activation potential compared with ERα; the mechanism underlying this difference is unclear. Here, we report that the binding of steroid receptor coactivator-1 (SRC-1) to the AF-1 domain of ERα is essential but not sufficient to facilitate synergy between the AF-1 and AF-2 domains, which is required for a full agonistic response to estradiol (E2). Complete synergy is achieved through the distinct hinge domain of ERα, which enables combined action of the AF-1 and AF-2 domains. AF-1 of ERβ lacks the capacity to interact with SRC-1, which prevents hinge-mediated synergy between AF-1 and AF-2, thereby explaining the reduced E2-mediated transactivation of ERβ. Transactivation of ERβ by E2 requires only the AF-2 domain. A weak agonistic response to tamoxifen occurs for ERα, but not for ERβ, and depends on AF-1 and the hinge-region domain of ERα.
The Gab1 scaffold regulates RTK-dependent dorsal ruffle formation through the adaptor Nck
Authors: Abella Jasmine V. , Vaillancourt Richard , Frigault Melanie M. , Ponzo Marisa G. , Zuo Dongmei , Sangwan Veena , Larose Louise , Park Morag
Published: March 2010

The polarised distribution of signals downstream from receptor tyrosine kinases (RTKs) regulates fundamental cellular processes that control cell migration, growth and morphogenesis. It is poorly understood how RTKs are involved in the localised signalling and actin remodelling required for these processes. Here, we show that the Gab1 scaffold is essential for the formation of a class of polarised actin microdomain, namely dorsal ruffles, downstream from the Met, EGF and PDGF RTKs. Gab1 associates constitutively with the actin-nucleating factor N-WASP. Following RTK activation, Gab1 recruits Nck, an activator of N-WASP, into a signalling complex localised to dorsal ruffles. Formation of dorsal ruffles requires interaction between Gab1 and Nck, and also requires functional N-WASP. Epithelial cells expressing Gab1ΔNck (Y407F) exhibit decreased Met-dependent Rac activation, fail to induce dorsal ruffles, and have impaired cell migration and epithelial remodelling. These data show that a Gab1-Nck signalling complex interacts with several RTKs to promote polarised actin remodelling and downstream biological responses.
Insulin regulates alveolar epithelial function by inducing Na+/K+-ATPase translocation to the plasma membrane in a process mediated by the action of Akt
Authors: Comellas Alejandro P. , Kelly Aileen M. , Trejo Humberto E. , Briva Arturo , Lee Joyce , Sznajder Jacob I. , Dada Laura A.
Published: March 2010

Stimulation of Na+/K+-ATPase translocation to the cell surface increases active Na+ transport, which is the driving force of alveolar fluid reabsorption, a process necessary to keep the lungs free of edema and to allow normal gas exchange. Here, we provide evidence that insulin increases alveolar fluid reabsorption and Na+/K+-ATPase activity by increasing its translocation to the plasma membrane in alveolar epithelial cells. Insulin-induced Akt activation is necessary and sufficient to promote Na+/K+-ATPase translocation to the plasma membrane. Phosphorylation of AS160 by Akt is also required in this process, whereas inactivation of the Rab GTPase-activating protein domain of AS160 promotes partial Na+/K+-ATPase translocation in the absence of insulin. We found that Rab10 functions as a downstream target of AS160 in insulin-induced Na+/K+-ATPase translocation. Collectively, these results suggest that Akt plays a major role in Na+/K+-ATPase intracellular translocation and thus in alveolar fluid reabsorption.
Combination of Metaheuristic and Exact Algorithms for Solving Set Covering-Type Optimization Problems
Authors: Muter İbrahim , Birbil Ş. İlker , Şahin Güvenç
Published: March 2010

We propose a new generic framework for solving combinatorial optimization problems that can be modeled as a set covering problem. The proposed algorithmic framework combines metaheuristics with exact algorithms through a guiding mechanism based on diversification and intensification decisions. After presenting this generic framework, we extensively demonstrate its application to the vehicle routing problem with time windows. We then conduct a thorough computational study on a set of well-known test problems, where we show that the proposed approach not only finds solutions that are very close to the best-known solutions reported in the literature, but also improves them. We finally set up an experimental design to analyze the effects of different parameters used in the proposed algorithm.
Vasopressin Antagonists: Pharmacotherapy for the Treatment of Heart Failure
Authors: Reilly Timothy , Schork Michelle R
Published: March 2010

Objective

To evaluate acute hemodynamic, short-term, and long-term effects of vasopressin antagonists in patients with heart failure (HF).

Data sources

Searches of the PubMed database (1966–February 2010) were conducted. Search terms included AVP receptor antagonist, CHF, tolvaptan, conivaptan, lixivaptan, HF, and hyponatremia. Manufacturers’ prescribing information, manufacturer Web site searches, and searches made on www.clinicaltrials.gov were also included.

Study selection and data extraction

All clinical trials identified from the reference search and data sources were reviewed. Articles were included if they were relevant to short-term and long-term outcomes of patients with HF who were treated with vasopressin antagonists.

Data synthesis

Trials of conivaptan, tolvaptan, and lixivaptan were evaluated. The evidence indicates that all agents increase urine output >10 mL/h, and conivaptan and tolvaptan decrease pulmonary capillary wedge pressure (–2.6 ± 0.7, –5.4 ± 0.7, and –4.6 ± 0.7 mm Hg for placebo, conivaptan 20 mg, and conivaptan 40 mg, respectively; –5.67 ± 4.58 to –6.38 ± 4.12 mm Hg for tolvaptan, and –4.16 ±4.57 mm Hg for placebo) in patients with HF. Both of these changes occur without inducing electrolyte abnormalities or renal dysfunction. Trials with conivaptan in acute HF have not demonstrated a benefit in cardiac index, mean arterial pressure, or vascular resistance. Data from clinical trials indicate that tolvaptan may decrease dyspnea (p < 0.05) and pedal edema (p < 0.001). To date, no large-scale trials of any agent have demonstrated an improvement in left ventricular systolic function, rehospitalization, worsening HF, or death.

Conclusions

Vasopressin antagonists cannot be considered routine pharmacotherapy for HF, Further, conivaptan should not be used for the treatment of acute HF. There is not enough literature to advocate for or against the use of lixivaptan in patients with HF. Tolvaptan may be considered for the treatment of hyponatremia.
Konstruktionen in der kommunikativen Praxis. Zur Notwendigkeit einer interaktionalen Anreicherung konstruktionsgrammatischer Ansätze
Authors: Günthner Susanne
Published: March 2010

The following contribution is dedicated to questions about the interface between interactionally based studies on the grammar of everyday language use and “usage-based” approaches from Construction Grammar.

Studies of grammatical structures in interactional use make it clear that concepts of grammar which assume homogeneous, static categories are only partly suitable for the description of spoken language, since grammatical structures in communicative practice are more heterogeneous, open and context dependent than postulated. At the same time, these analyses show that prefabricated patterns form an important means of solving communicative tasks precisely in oral communication, which takes place under time pressure. These factors make usage-based approaches from Construction Grammar attractive as a framework for a practically oriented examination of grammar, though many divergences exist all the same.

On the basis of empirical analyses of pseudo-clefts and wenn constructions in spoken German, the article discusses the possibility of extending usage-based approaches from Construction Grammar, which aims to integrate the circumstances of language production and reception in communicative practice into the analysis, and therefore the fact that we use language in order to carry out social activities in the process of interaction.