bluuebunny/crossref_metadata_2025 huggingface dataset:
‘Golden Age’ versus ‘Bad Old Days’
Authors: Locke Abigail , Horton-Salway Mary
Published: May 2010

Childbirth is seen as a medical event, and pregnancy, a time when parents-to-be are in need of advice. This article provides a discursive analysis of how such advice is given in antenatal classes. Using audio-recorded data from National Childbirth Trust (NCT) antenatal classes, we analyse how class leaders talk to class members about pregnancy, childbirth and infant care. We identify a pattern of advice giving in which class leaders construct ‘golden age’ or ‘bad old days’ stories variably to contrast the practices of the past (‘then’) with current practices (‘now’). These contrasting repertoires operate against a backdrop of medicalization and societal expectations that are both current and out-dated, providing a constitutive framework to support class leaders’ evaluations and advice on pregnancy, childbirth and infant care.
Political leanings vary with facial expression processing and psychosocial functioning
Authors: Vigil Jacob M.
Published: May 2010

Conservative, Republican sympathizers show heightened threat reactivity, but greater felt happiness than liberal, Democrat sympathizers. Recent evolutionary models interpret these findings in the context of broader perceptual and expressive proclivities for advertising cues of competency (Republicans) and trustworthiness (Democrats) to others, and in ways that facilitate the formation of distinct social networks, in coordination with individuals’ life histories. Consistent with this perspective, I found that Republican sympathizers were more likely to report larger social networks and interpret ambiguous facial stimuli as expressing more threatening emotions as compared to Democrat sympathizers, who also reported greater emotional distress, relationship dissatisfaction, and experiential hardships. The findings are discussed in the context of proximate and ultimate explanations of social cognition, relationship formation, and societal cohesion.
Do the ideological beliefs of peers predict the prejudiced attitudes of other individuals in the group?
Authors: Poteat V. Paul , Spanierman Lisa B.
Published: May 2010

The authors used multilevel modeling to examine whether peer group ideological beliefs ( n = 109 friendship groups) predicted the homophobic and racist attitudes of other individuals within the group ( n = 395 college students). Results indicated that the social dominance orientation (SDO), right-wing authoritarianism (RWA), and universal-diverse orientation (UDO) ideological beliefs of peers predicted the prejudiced attitudes of other group members, over and above individuals’ own ideological views. Additionally, the strength with which individuals’ own ideological beliefs predicted their prejudiced attitudes varied systematically across peer groups. Affiliations with high-RWA peers strengthened the extent to which individuals’ own SDO and RWA predicted their prejudiced attitudes. Results suggest the ideological beliefs of peers are relevant to predicting the prejudiced attitudes of the individuals with whom they affiliate. Although specific peer ideologies differentially predicted forms of prejudice, the overall contribution of these peer ideology beliefs to the prediction of individuals’ prejudiced attitudes was comparable for both homophobic and racist attitudes. Attention to proximal social networks and the social dynamics within these networks can contribute to better explanations of individual differences in prejudiced attitudes.
Acute hyperhomocysteinaemia blunts endothelial dependent and endothelial independent vasodilatation in diabetic patients
Authors: Chousos Ilias , Perrea Despoina , Kyriaki Despoina , Liatis Stavros , Katsilambros Nicholas , Makrilakis Konstantinos
Published: May 2010

The aim of this study was to investigate the effects of acute, methionine-induced hyperhomocysteinaemia on endothelial function of skin microcirculation in diabetic compared with non-diabetic persons. Endothelial- and non-endothelial-dependent vasodilatation of the skin microcirculation were evaluated in 20 persons with type 2 diabetes (59.95±2.36 years old) and 24 age- and gender-matched controls using laser Doppler flowmetry, during fasting and four hours after 0.1 g/kg methionine-induced acute hyperhomocysteinaemia. Methionine loading caused a comparable increase of homocysteine in the two groups. Four hours after induction of acute hyperhomocysteinaemia, endothelial-dependent vasodilatation (acetylcholine effect) was lower in the diabetic patients compared with the controls ([mean±SEM] 8.09±1.18-fold increase in diabetic patients vs. 11.11±1.33-fold increase in controls, p=0.027). The same was observed for the endothelial-independent vasodilatation (sodium-nitroprusside effect, 7.55±0.80-fold increase in the diabetic patients vs. 12.19±1.26-fold increase in controls, p=0.008). Acute hyperhomocysteinaemia causes a decreased response of both endothelial-dependent and endothelial-independent vasodilatation of the skin microcirculation in diabetic compared with non-diabetic persons. These alterations might contribute to the vascular complications of hyperhomocysteinaemia in diabetes, and especially the microvascular ones.
Tissue-specific effects of valsartan on rstn and fiaf gene expression in the ob/ob mouse
Authors: Imran SA , Brown RE , Wilkinson M
Published: May 2010

The RAS is a novel target in the study of diabetes, and clinical trials have indicated that ARBs, such as valsartan, may exert some of their clinical effects through an influence on adipose tissue. We studied the effect of valsartan on adipokine genes resistin ( rstn) and fasting-induced adipose factor ( fiaf) using obese and diabetic ob/ob mice. In addition to visceral and subcutaneous fat, rstn and fiaf mRNA levels were also measured in several other tissues known to express these adipokines, including the pituitary, cerebral cortex and hypothalamus. The significant findings were that (a) fiaf gene expression was elevated two- to fourfold in visceral and subcutaneous fat from ob/ob mice, compared with lean controls; (b) the increase in fiaf mRNA in subcutaneous, but not visceral, fat from ob/ob mice was returned to lean control levels following 2 weeks of valsartan treatment; (c) fiaf expression was reduced in the hypothalamus, but not in the cortex or pituitary, of ob/ob mice; (d) rstn expression was greatly reduced in visceral fat from ob/ob mice, compared with lean controls, but this was unaffected by valsartan; and (e) rstn expression was unchanged in all other tissues from ob/ob mice, with or without valsartan treatment.
Law and the Foucauldian Wild West in Michael Cimino’s Heaven’s Gate
Authors: Young Diana
Published: May 2010

Classic western films often conceive of the west as existing in a legal void, where the central conflict is a binary one between lawlessness and legalization. The law is a monolith, and the legalization process is linear — a narrative of the west’s inexorable evolution toward a modern state governed by the rule of law. Cimino’s Heaven’s Gate presents a more postmodernist, pluralist conception. There is no grand narrative of legalization; the film envisages a discourse of justice emerging from the interaction of a variety of discourses, and which appears to be a unity only from the vantage point of history.
Subversives All! Ronald Reagan & the Paternal Roots of “Law and Order” at Home and Abroad
Authors: Dudas Jeffrey
Published: May 2010

Law and society scholars have identified the call for “law and order” during the Cold War-era as a method for opposing egalitarian social change. But the law and order narrative was not simply a coded phrase meant to appeal to the anxieties of traditionally-powerful Americans. Instead, the depiction of egalitarian politics consistently offered in law and order narratives – that such politics was the product of unruly children who, due to a chronic lack of paternal influence, were deficient in the prototypical American virtue of self-discipline and, so, were vulnerable to communist indoctrination – performed constitutive work. The law and order narrative emphasized, first, that egalitarian politics was subversive of the American nation. Second, it offered a counter-subversive response to egalitarian politics that infantilized marginalized people and recommended the exercise of a stern, frequently violent, fatherly authority that would impart to them the discipline necessary for productive American citizenship. Exploration of Ronald Reagan’s employment of the law and order narrative to oppose student protestors while Governor of California and the Sandinista regime in Nicaragua while President of the United States exposes the paternal roots of American law and order both at home and abroad.
Living well and sexual self-determination: Expanding human rights discourse about sex and sexuality
Authors: Weinman Michael
Published: May 2010

This work argues that a reformulated capability approach expands the horizons established in earlier and contemporary classical rights-based approaches to gay rights and reproductive rights, by raising the threshold from recognizing an individual’s right to non-interference against the state or other entities to recognizing a shared responsibility to support everyone’s capacity to fulfill their potential. The “right to choose” thus comes to be cast not a right to be left alone, but as a social responsibility to provide for each individual’s fulfillment of our ergon, usually “function,” but really more like one’s work in being alive.
Sacrifice and the Singular
Authors: Slaughter Marty
Published: May 2010

One kind of sacrifice the law undergoes is that of the singular. As discussed by Levinas, Derrida, Agamben and Johann van der Walt, ethics and law must apply norms and rules that are general to the absolutely unique individual, his particular acts and their circumstances. These are subsumed under a rule which at the same time limits their unique characteristics, i.e., sacrifices their particularity. To some extent the law acknowledges this problem and attempts to recuperate the singular through the concept and application of equity. This is, however, never completely successful; there is always a remainder of the singular that is sacrificed. Sacrifice is a very part of the law.
Cuts, Flows, and the Geographies of Property
Authors: Blomley Nicholas
Published: May 2010

How is property geographical? The making of liberal property, I argue, relies upon a topographical logic, premised on the production of bounded, coherent spaces, through which the individuated subjects and objects of property can be rendered legible. Such a spatialization helps sustain the territorialization of property, in which the government of space becomes a means for the enactment of property. The production of such spaces requires conscious ‘cuts’ in the processual networks through which social spaces are produced. As such, property should be seen as a conditional achievement, ever threatened by unwanted relationality and boundary crossing. I draw from Kate Grenville’s novel The Secret River to explore property’s spaces, and their ambivalent ethical and practical work.
Paracellular permeability is increased by basal lipopolysaccharide in a primary culture of colonic epithelial cells; an effect prevented by an activator of Toll-like receptor-2
Authors: Hanson Peter J. , Moran Anthony P. , Butler Kate
Published: May 2010

Lipopolysaccharide (LPS), which generally activates Toll-like receptor 4 (TLR4), is expressed on commensal colonic bacteria. In a number of tissues, LPS can act directly on epithelial cells to increase paracellular permeability. Such an effect in the colon would have an important impact on the understanding of normal homeostasis and of pathology. Our aim was to use a novel primary culture of colonic epithelial cells grown on Transwells to investigate whether LPS, or Pam3CSK4, an activator of TLR2, affected paracellular permeability. Consequently, [14C]-mannitol transfer and transepithelial electrical resistance (TEER) were measured. The preparation consisted primarily of cytokeratin-18 positive epithelial cells that produced superoxide, stained for mucus with periodic acid-Schiff reagent, exhibited alkaline phosphatase activity and expressed TLR2 and TLR4. Tight junctions and desmosomes were visible by transmission electron microscopy. Basally, but not apically, applied LPS from Escherichia coli increased the permeability to mannitol and to a 10-kDa dextran, and reduced TEER. The LPS from Helicobacter pylori increased paracellular permeability of gastric cells when applied either apically or basally, in contrast to colon cells, where this LPS was active only from the basal aspect. A pan-caspase inhibitor prevented the increase in caspase activity caused by basal E. coli LPS, and reduced the effects of LPS on paracellular permeability. Synthetic Pam3CSK4in the basal compartment prevented all effects of basal E. coli LPS. In conclusion, LPS applied to the base of the colonic epithelial cells increased paracellular permeability by a mechanism involving caspase activation, suggesting a process by which perturbation of the gut barrier could be exacerbated. Moreover, activation of TLR2 ameliorated such effects.
Hemin and a metabolic derivative coprohemin modulate the TLR4 pathway differently through different molecular targets
Authors: Piazza Matteo , Damore Gaetana , Costa Barbara , Gioannini Theresa L. , Weiss Jerrold P. , Peri Francesco
Published: May 2010

Heme is a prosthetic group in a large number of essential proteins that have a pivotal role in oxygen transport, storage and electron shuttling. High amounts of free heme are associated with pathological states. Recently, it has been suggested that activation of Toll-like receptor 4 (TLR4) is one of the ways in which the ‘danger signal’ of free heme is detected. Here, we examine the biochemical basis of the modulation of the TLR4 pathway by hemin (iron(III)-protoporphyrin IX) and its metabolic, oxidated derivative coprohemin (iron(III)-coproporphyrin I). High concentrations of hemin (50 μM) triggered TLR4-mediated IL-8 production in the human HEK293/TLR4 cell line in the absence of the co-receptors CD14 and MD-2; the latter an essential co-receptor for TLR4 activation by endotoxin. Hemin and endotoxin have additive effects when co-administrated to HEK/TLR4 cells, suggesting that hemin and endotoxin activate TLR4 by different mechanisms. Coprohemin, in contrast to hemin, is unable to trigger TLR4-dependent activation of HEK/TLR4 cells, but instead causes dose-dependent inhibition of endotoxin-stimulated IL-8 production. The inhibitory effect of coprohemin is paralleled by reduced delivery of endotoxin to MD-2 (-TLR4) that is necessary for activation of TLR4 by endotoxin. Thus, despite their similar chemical structure, hemin and coprohemin have very different effects on the TLR4 pathway, the former acting as a mild agonist of TLR4, the latter as an antagonist selectively targeting the endotoxin—MD-2 interaction.
CD14 (C-260T) polymorphism is not associated with sudden infant death syndrome (SIDS) in a large South Australian cohort
Authors: Highet Amanda R. , Gibson Catherine S. , Goldwater Paul N.
Published: May 2010

Similarities have been drawn between models of endotoxic shock and gross and microscopic pathology observed in sudden infant death syndrome (SIDS) cases. Polymorphisms in genes that influence the expression of endotoxin receptors could affect the outcome of toxaemia, and could, therefore, play a role in SIDS. The CD14 gene promoter contains a single nucleotide polymorphism that affects the level of CD14 gene expression. The TT genotype of the CD14 (C-260T) polymorphism causes a significantly higher density of CD14 receptor expression on monocytes which makes the individual more sensitive to endotoxin than those with the wild-type (CC). This investigation was designed to determine whether SIDS infants have a higher frequency of the CD14 (C-260T) polymorphism compared with non-SIDS controls. One hundred and sixteen SIDS and 228 control infants were genotyped using PCR followed by restriction fragment length analysis of amplified product. Carriage of the TT or CT genotypes did not significantly differ between SIDS and control infants (P = 0.218 and 0.081, respectively). The frequencies observed in the control group were consistent with Hardy—Weinberg equilibrium and did not differ significantly from the published frequencies in Caucasian Australians. These results suggest that CD14 (C-260T) polymorphism is unlikely to be implicated in SIDS.
The Future of Emotion Research in the Study of Psychopathology
Authors: Kring Ann M.
Published: May 2010

Research on emotion and psychopathology has blossomed due in part to the translation of affective science theory and methods to the study of diverse disorders. This translational approach has helped the field to hone in more precisely on the nature of emotion deficits to identify antecedent causes and maintaining processes, and to develop promising new interventions. The future of emotion research in psychopathology will benefit from three inter-related areas, including an emphasis on emotion difficulties that cut across traditional diagnostic boundaries (i.e., a transdiagnostic approach), the explicit linking of emotion and cognition in behavioral and neuroimaging studies in psychopathology, and continued translation of the latest conceptualizations of emotion to the study of psychopathology.
The development of a new technique for the decontamination of trans-oesophageal echocardiography probes
Authors: Richardson Andrew , Cowan Charlie , Waters Jan , Diprose Paul
Published: May 2010

Decontamination of trans-oesophageal echocardiography (TOE) probes to a validated standard is a troublesome area, since their electrical components prohibit full immersion. We report the development of a novel device designed to keep the electrical components of the TOE probe dry while allowing the rest of the probe to be subjected to a validated decontamination process.
Effect of the dried residues of two hand gels on the survival of meticillin-resistant Staphylococcus aureus and Acinetobacter calcoaceticus-baumannii
Authors: Hall Tony J , Wren Michael WD , Wareham David W , Jeanes Annette , Gant Vanya A
Published: May 2010

We assessed the ability of the residues of an alcohol-based hand gel (hand gel A) and an Aloe vera-based CuAL42 copper biocide-containing hand gel (hand gel B), to support the survival of meticillin-resistant Staphylococcus aureus (MRSA) and Acinetobacter calcoaceticus-baumannii (ACCB). One-millilitre samples of hand gels A and B were spread over marked 20 cm2 areas and dried overnight. MRSA or ACCB (1 × 104 CFU) were spread onto the dried residue and contact plated at various times. MRSA and ACCB survived for 8 hours on hand gel A residue, whilst MRSA did not survive on hand gel B residue and ACCB survived less than 30 min. Low concentrations of hand gel A facilitate the growth of ACCB, but this is not the case for hand gel B. Hand gel A is extensively used in UK hospitals and its residue left on high-touch surfaces may support the survival of bacteria that cause healthcare-acquired infections.
Lifting the lid: a clinical audit on commode cleaning
Authors: Bucior Helen , Cochrane Joan
Published: May 2010

Many healthcare-associated infections (HCAIs) are preventable by infection control procedures designed to interrupt the transmission of organisms from a source. Commodes are in use constantly throughout healthcare facilities. Therefore commode surfaces are constantly handled, and any pathogens present have the potential to be transferred to not only other surfaces but also, more importantly, to patients, thus compromising patient safety. In order to examine the effectiveness and thoroughness of cleaning commodes an audit was undertaken to assess compliance with evidence-based practice. This audit demonstrates a cycle which includes defining best practice, implementing best practice, monitoring best practice and taking action to improve practice. The audit results confirmed an issue that the authors had long suspected. That is, that commodes allocated to individual patients are not always cleaned after every use. Using adenosine triphosphate (ATP) bioluminescence as an indicator of organic soiling also demonstrated that commodes that were considered clean were not always cleaned to a high standard. Implementing the audit recommendations improves staff knowledge through education, standardises cleaning procedures and ultimately improves patient safety.
Critical Conversations
Authors: Rizk Dahlia , Greeno Ron , Maldonado Lawrence , Limenes Lydia , Stewart Samantha , Wittnebel Karl , Killu Claude , Hackner Dani
Published: May 2010

Vignette: An 88-year-old woman visiting from out of town, and out of network, presents to the emergency department with chest pain and rapidly progressing. Delays in the emergency department due to bed capacity lead the patient to be placed under the care of a hospitalist and emergency department physician while waiting for a floor bed. She develops increasing hemodynamic instability and is “upgraded” to an ICU bed with a presumptive diagnosis of septic shock. The ICU team is called.
Leukemia regression by vascular disruption and antiangiogenic therapy
Authors: Madlambayan Gerard J. , Meacham Amy M. , Hosaka Koji , Mir Saad , Jorgensen Marda , Scott Edward W. , Siemann Dietmar W. , Cogle Christopher R.
Published: May 2010

Acute myelogenous leukemias (AMLs) and endothelial cells depend on each other for survival and proliferation. Monotherapy antivascular strategies such as targeting vascular endothelial growth factor (VEGF) has limited efficacy in treating AML. Thus, in search of a multitarget antivascular treatment strategy for AML, we tested a novel vascular disrupting agent, OXi4503, alone and in combination with the anti-VEGF antibody, bevacizumab. Using xenotransplant animal models, OXi4503 treatment of human AML chloromas led to vascular disruption in leukemia cores that displayed increased leukemia cell apoptosis. However, viable rims of leukemia cells remained and were richly vascular with increased VEGF-A expression. To target this peripheral reactive angiogenesis, bevacizumab was combined with OXi4503 and abrogated viable vascular rims, thereby leading to enhanced leukemia regression. In a systemic model of primary human AML, OXi4503 regressed leukemia engraftment alone and in combination with bevacizumab. Differences in blood vessel density alone could not account for the observed regression, suggesting that OXi4503 also exhibited direct cytotoxic effects on leukemia cells. In vitro analyses confirmed this targeted effect, which was mediated by the production of reactive oxygen species and resulted in apoptosis. Together, these data show that OXi4503 alone is capable of regressing AML by a multitargeted mechanism and that the addition of bevacizumab mitigates reactive angiogenesis.
Serum ferritin is derived primarily from macrophages through a nonclassical secretory pathway
Authors: Cohen Lyora A. , Gutierrez Lucia , Weiss Avital , Leichtmann-Bardoogo Yael , Zhang De-liang , Crooks Daniel R. , Sougrat Rachid , Morgenstern Avigail , Galy Bruno , Hentze Matthias W. , Lazaro Francisco J. , Rouault Tracey A. , Meyron-Holtz Esther G.
Published: May 2010

The serum ferritin concentration is a clinical parameter measured widely for the differential diagnosis of anemia. Its levels increase with elevations of tissue iron stores and with inflammation, but studies on cellular sources of serum ferritin as well as its subunit composition, degree of iron loading and glycosylation have given rise to conflicting results. To gain further understanding of serum ferritin, we have used traditional and modern methodologies to characterize mouse serum ferritin. We find that both splenic macrophages and proximal tubule cells of the kidney are possible cellular sources for serum ferritin and that serum ferritin is secreted by cells rather than being the product of a cytosolic leak from damaged cells. Mouse serum ferritin is composed mostly of L-subunits, whereas it contains few H-subunits and iron content is low. L-subunits of serum ferritin are frequently truncated at the C-terminus, giving rise to a characteristic 17-kD band that has been previously observed in lysosomal ferritin. Taken together with the fact that mouse serum ferritin is not detectably glycosylated, we propose that mouse serum ferritin is secreted through the nonclassical lysosomal secretory pathway.
Interferon-γ and tumor necrosis factor-α induce an immunoinhibitory molecule, B7-H1, via nuclear factor-κB activation in blasts in myelodysplastic syndromes
Authors: Kondo Asaka , Yamashita Taishi , Tamura Hideto , Zhao Wanhong , Tsuji Takashi , Shimizu Masumi , Shinya Eiji , Takahashi Hidemi , Tamada Koji , Chen Lieping , Dan Kazuo , Ogata Kiyoyuki
Published: May 2010

During disease progression in myelodysplastic syndromes (MDS), clonal blasts gain a more aggressive nature, whereas nonclonal immune cells become less efficient via an unknown mechanism. Using MDS cell lines and patient samples, we showed that the expression of an immunoinhibitory molecule, B7-H1 (CD274), was induced by interferon-γ (IFNγ) and tumor necrosis factor-α (TNFα) on MDS blasts. This induction was associated with the activation of nuclear factor-κB (NF-κB) and nearly completely blocked by an NF-κB inhibitor, pyrrolidine dithiocarbamate (PDTC). B7-H1+ MDS blasts had greater intrinsic proliferative capacity than B7-H1− MDS blasts when examined in various assays. Furthermore, B7-H1+ blasts suppressed T-cell proliferation and induced T-cell apoptosis in allogeneic cocultures. When fresh bone marrow samples from patients were examined, blasts from high-risk MDS patients expressed B7-H1 molecules more often compared with those from low-risk MDS patients. Moreover, MDS T cells often overexpressed programmed cell death 1 (PD-1) molecules that transmit an inhibitory signal from B7-H1 molecules. Taken together, these findings provide new insight into MDS pathophysiology. IFNγ and TNFα activate NF-κB that in turn induces B7-H1 expression on MDS blasts. B7-H1+ MDS blasts have an intrinsic proliferative advantage and induce T-cell suppression, which may be associated with disease progression in MDS.
EGR-1 activation by EGF inhibits MMP-9 expression and lymphoma growth
Authors: Bouchard Frédéric , Bélanger Simon D. , Biron-Pain Katherine , St-Pierre Yves
Published: May 2010

Progression of hematologic malignancies is strongly dependent on bidirectional interactions between tumor cells and stromal cells. Expression of members of the matrix metalloproteinase (MMP) family by stromal cells is a central event during these interactions. However, although several studies have focused on the mechanisms responsible for induction of MMP in stromal cells, the signals that negatively regulate their secretion of in these cells remain largely unknown. Here, we provide evidence that MMP-9 production by stromal cells is suppressed through activation of early growth response protein 1 (EGR-1), thereby inhibiting the growth of thymic lymphoma. We found that EGR-1 expression is induced in stromal cells after contact with lymphoma cells via epidermal growth factor (EGF). Moreover, development of thymic lymphoma was inhibited when induced by lymphoma cells overexpressing EGF compared with control lymphoma cells. Using transgenic mice containing MMP-9 promoter-driven luciferase transgene in its genome, we further demonstrated that EGF/EGR-1 repressed transcriptional activation of the MMP-9 gene by stromal cells. De novo expression of EGR-1 alone by gene transfer or exposure to recombinant human EGF also inhibited MMP-9 expression. Taken together, these results indicate that EGR-1 could be a source of novel targets for therapeutic intervention in lymphoid tumors in which MMP-9 plays a critical role.
Myeloproliferative neoplasm induced by constitutive expression of JAK2V617F in knock-in mice
Authors: Marty Caroline , Lacout Catherine , Martin Antoine , Hasan Salma , Jacquot Sylvie , Birling Marie-Christine , Vainchenker William , Villeval Jean-Luc
Published: May 2010

The Jak2V617F mutation is found in most classical BCR/ABL-negative myeloproliferative neoplasms (MPNs). Usually, heterozygosity of the mutation is associated with essential thrombocythemia (ET) and homozygosity with polycythemia vera (PV). Retrovirally transduced or transgenic animal models have shown that the mutation is sufficient for MPN development but that the level of expression is crucial for MPN phenotypes. Therefore we investigated the effect of an endogenous heterozygous expression of Jak2V617F in knock-in (KI) mice. These animals displayed constitutive JAK2 activation and autonomous erythroid progenitor cell growth. Mice suffered from marked polycythemia, granulocytosis and thrombocytosis. Spleens and marrows displayed myeloid trilineage hyperplasia. Most animals survived to develop advanced fibrosis in these organs at around 9 months of age. In conclusion, constitutive heterozygous expression of JAK2V617F in mice is not embryo-lethal but results in severe PV-like disease with secondary myelofibrosis and not in ET-like disease as expected from patient study.
Elucidation of the integrin LFA-1–mediated signaling pathway of actin polarization in natural killer cells
Authors: Mace Emily M. , Zhang Jinyi , Siminovitch Katherine A. , Takei Fumio
Published: May 2010

The leukocyte integrin LFA-1 is critical for natural killer (NK) cell cytotoxicity as it mediates NK-cell adhesion to target cells and generates activating signals that lead to polarization of the actin cytoskeleton. However, the LFA-1–mediated signaling pathway is not fully understood. Here, we examined the subcellular localization of actin-associated proteins in wild-type, talin-deficient, and Wiskott-Aldrich Syndrome protein (WASP)–deficient NK cells bound to beads coated with the LFA-1 ligand intercellular adhesion molecule-1 (ICAM-1). In addition, we carried out coimmunoprecipitation analyses and also used a pharmacologic reagent to reduce the level of phosphatidylinositol-4,5-bisphosphate (PIP2). The results revealed the following signaling pathways. Upon ICAM-1 binding to LFA-1, talin redistributes to the site of LFA-1 ligation and initiates 2 signaling pathways. First, talin recruits the actin nucleating protein complex Arp2/3 via constitutive association of vinculin with talin and Arp2/3. Second, talin also associates with type I phosphatidylinositol 4-phosphate 5-kinase (PIPKI) and binding of LFA-1 to ICAM-1 results in localized increase in PIP2. This increase in PIP2 recruits WASP to the site of LFA-1 ligation where WASP promotes Arp2/3-mediated actin polymerization. These processes are critical for the initiation of NK cell–mediated cytotoxicity.
B7-H1/CD80 interaction is required for the induction and maintenance of peripheral T-cell tolerance
Authors: Park Jang-June , Omiya Ryusuke , Matsumura Yumiko , Sakoda Yukimi , Kuramasu Atsuo , Augustine Mathew M. , Yao Sheng , Tsushima Fumihiko , Narazaki Hidehiko , Anand Sudarshan , Liu Yingjia , Strome Scott E. , Chen Lieping , Tamada Koji
Published: May 2010

T-cell tolerance is the central program that prevents harmful immune responses against self-antigens, in which inhibitory PD-1 signal given by B7-H1 interaction plays an important role. Recent studies demonstrated that B7-H1 binds CD80 besides PD-1, and B7-H1/CD80 interaction also delivers inhibitory signals in T cells. However, a role of B7-H1/CD80 signals in regulation of T-cell tolerance has yet to be explored. We report here that attenuation of B7-H1/CD80 signals by treatment with anti–B7-H1 monoclonal antibody, which specifically blocks B7-H1/CD80 but not B7-H1/PD-1, enhanced T-cell expansion and prevented T-cell anergy induction. In addition, B7-H1/CD80 blockade restored Ag responsiveness in the previously anergized T cells. Experiments using B7-H1 or CD80-deficient T cells indicated that an inhibitory signal through CD80, but not B7-H1, on T cells is responsible in part for these effects. Consistently, CD80 expression was detected on anergic T cells and further up-regulated when they were re-exposed to the antigen (Ag). Finally, blockade of B7-H1/CD80 interaction prevented oral tolerance induction and restored T-cell responsiveness to Ag previously tolerized by oral administration. Taken together, our findings demonstrate that the B7-H1/CD80 pathway is a crucial regulator in the induction and maintenance of T-cell tolerance.
Selective abrogation of the uPA-uPAR interaction in vivo reveals a novel role in suppression of fibrin-associated inflammation
Authors: Connolly Brian M. , Choi Eun Young , Gårdsvoll Henrik , Bey Alexandra L. , Currie Brooke M. , Chavakis Triantafyllos , Liu Shihui , Molinolo Alfredo , Ploug Michael , Leppla Stephen H. , Bugge Thomas H.
Published: May 2010

The urokinase plasminogen activator receptor (uPAR) has emerged as a potential regulator of cell adhesion, cell migration, proliferation, differentiation, and cell survival in multiple physiologic and pathologic contexts. The urokinase plasminogen activator (uPA) was the first identified ligand for uPAR, but elucidation of the specific functions of the uPA-uPAR interaction in vivo has been difficult because uPA has important physiologic functions that are independent of binding to uPAR and because uPAR engages multiple ligands. Here, we developed a new mouse strain (PlauGFDhu/GFDhu) in which the interaction between endogenous uPA and uPAR is selectively abrogated, whereas other functions of both the protease and its receptor are retained. Specifically, we introduced 4 amino acid substitutions into the growth factor domain (GFD) of uPA that abrogate uPAR binding while preserving the overall structure of the domain. Analysis of PlauGFDhu/GFDhu mice revealed an unanticipated role of the uPA-uPAR interaction in suppressing inflammation secondary to fibrin deposition. In contrast, leukocyte recruitment and tissue regeneration were unaffected by the loss of uPA binding to uPAR. This study identifies a principal in vivo role of the uPA-uPAR interaction in cell-associated fibrinolysis critical for suppression of fibrin accumulation and fibrin-associated inflammation and provides a valuable model for further exploration of this multifunctional receptor.
The human collagen beta(1-O)galactosyltransferase, GLT25D1, is a soluble endoplasmic reticulum localized protein
Authors: Liefhebber Jolanda MP , Punt Simone , Spaan Willy JM , van Leeuwen Hans C
Published: May 2010

Background

Glycosyl transferases transfer glycosyl groups onto their substrate. Localization partially defines their function. Glycosyl transferase 25 domain 1 (GLT25D1) was recently shown to have galactosyltransferase activity towards collagens and another well known substrate, mannose binding lectin (MBL). To gain more insight in the role of galactosylation of lysines in the Gly-X-Lys repeats of collagenous proteins, we investigated the subcellular localization of GLT25D1.

Results

Immunofluorescence analysis of GLT25D1 expressed in the human hepatoma cell line (Huh7), revealed a perinuclear lattice like staining, resembling localization to the endoplasmic reticulum (ER). Possible targeting signals, an N-terminal signal sequence and a C-terminal ER-retention signal, were identified using prediction programs. These signals were then investigated by constructing a series of epitope-tagged forms of GLT25D1 that were analyzed by immunofluorescence and western blotting. In agreement with the predictions our results show that GLT25D1 is directed to the ER lumen as a soluble protein and retained there. Moreover, using two endoglycosidase enzymes EndoH and EndoF, we demonstrate that the putative bi-functional glycosyl transferase itself is a glycoprotein. Additionally we examined co-localization of GLT25D1 with MBL and lysyl hydroxylase 3 (LH3, PLOD3), which is a protein able to catalyze hydroxylation of lysine residues before they can be glycosylated. We demonstrate overlapping localization patterns of GLT25D1, MBL and LH3.

Conclusions

Taken together our data indicate that galactosylation of collagenous proteins by the soluble GLT25D1 occurs in the early secretory pathway.
A three-stage approach for genome-wide association studies with family data for quantitative traits
Authors: Chen Ming-Huei , Larson Martin G , Hsu Yi-Hsiang , Peloso Gina M , Guo Chao-Yu , Fox Caroline S , Atwood Larry D , Yang Qiong
Published: May 2010

Background

Genome-wide association (GWA) studies that use population-based association approaches may identify spurious associations in the presence of population admixture. In this paper, we propose a novel three-stage approach that is computationally efficient and robust to population admixture and more powerful than the family-based association test (FBAT) for GWA studies with family data.

We propose a three-stage approach for GWA studies with family data. The first stage is to perform linear regression ignoring phenotypic correlations among family members. SNPs with a first stage p-value below a liberal cut-off (e.g. 0.1) are then analyzed in the second stage that employs a linear mixed effects (LME) model that accounts for within family correlations. Next, SNPs that reach genome-wide significance (e.g. 10-6 for 34,625 genotyped SNPs in this paper) are analyzed in the third stage using FBAT, with correction of multiple testing only for SNPs that enter the third stage. Simulations are performed to evaluate type I error and power of the proposed method compared to LME adjusting for 10 principal components (PC) of the genotype data. We also apply the three-stage approach to the GWA analyses of uric acid in Framingham Heart Study's SNP Health Association Resource (SHARe) project.

Results

Our simulations show that whether or not population admixture is present, the three-stage approach has no inflated type I error. In terms of power, using LME adjusting PC is only slightly more powerful than the three-stage approach. When applied to the GWA analyses of uric acid in the SHARe project of FHS, the three-stage approach successfully identified and confirmed three SNPs previously reported as genome-wide significant signals.

Conclusions

For GWA analyses of quantitative traits with family data, our three-stage approach provides another appealing solution to population admixture, in addition to LME adjusting for genetic PC.
Identification of Mycobacterium tuberculosis clinical isolates in Bangladesh by a species distinguishable multiplex PCR
Authors: Nakajima Chie , Rahim Zeaur , Fukushima Yukari , Sugawara Isamu , van der Zanden Adri GM , Tamaru Aki , Suzuki Yasuhiko
Published: May 2010

Background

Species identification of isolates belonging to the Mycobacterium tuberculosis complex (MTC) seems to be important for the appropriate treatment of patients, since M. bovis is naturally resistant to a first line anti-tuberculosis (TB) drug, pyrazinamide, while most of the other MTC members are susceptible to this antimicrobial agent. A simple and low-cost differentiation method was needed in higher TB burden countries, such as Bangladesh, where the prevalence of M. bovis among people or cattle has not been investigated.

Methods

Genetic regions cfp32, RD9 and RD12 were chosen as targets for a species distinguishable multiplex PCR and the system was evaluated with twenty reference strains of mycobacterial species including non-tubercular mycobacteria (NTM). A total of 350 clinical MTC isolates obtained in Bangladesh were then analyzed with this multiplex PCR.

Results

All of the MTC reference strains gave expected banding patterns and no non-specific amplifications were observed in the NTM strains. Out of 350 clinical isolates examined by this method, 347 (99.1%) were positive for all of the cfp32, RD9 and RD12 and determined as M. tuberculosis. Two isolates lacked cfp32 PCR product and one lacked RD12, however, those three samples were further examined and identified as M. tuberculosis by the sequence analyses of hsp65 and gyrB.

Conclusions

The MTC-discrimination multiplex PCR (MTCD-MPCR) developed in this study showed high specificity and was thought to be very useful as a routine test because of its simplicity. In the current survey, all the 350 MTC isolates obtained from Bangladesh TB patients were determined as M. tuberculosis and no other MTC were detected. This result suggested the general TB treatment regimen including pyrazinamide to be the first choice in Bangladesh.
Common polymorphisms of calpain-10 and the risk of Type 2 Diabetes in a Tunisian Arab population: a case-control study
Authors: Ezzidi Intissar , Turki Amira , Messaoudi Safia , Chaieb Molka , Kacem Maha , Al-Khateeb Ghada M , Mahjoub Touhami , Almawi Wassim Y , Mtiraoui Nabil
Published: May 2010

Background

Genetic variations in the calpain-10 gene (CAPN10), in particular the at-risk diplotype (112/121), were previously implicated with increased risk of type 2 diabetes (T2D).

Methods

We examined the association of CAPN10 UCSNP-43 (rs3792267), UCSNP-19 (rs3842570), and UCSNP-63 (rs5030952) SNPs with T2D in 917 Tunisian T2D patients and 748 non-diabetic controls. CAPN10 genotyping was done by PCR-RFLP.

Results

Enrichment of UCSNP-19 2R (minor) allele and 2R/2R genotype was found in T2D patients; the allele and genotype distribution of UCSNP-43 and UCSNP-63 alleles and genotypes were not significantly different between patient groups and non-diabetic control subjects. Regression analysis demonstrated progressive increases in T2D risk in 3R/2R [OR (95% CI) = 1.35 (1.08 - 1.68)] and 2R/2R [OR (95% CI) = 1.61 (1.20 - 2.18)] genotypes. Of the six haplotypes detected, enrichment of haplotype 111 (UCSNP-43/UCSNP-19/UCSNP-63) was seen in patients (Pc = 0.034); the distribution of the other haplotypes was comparable between patients and control subjects; neither haplotype 211 nor haplotype 212 was observed. Furthermore, the frequency of all CAPN10 diplotypes identified, including the "high-risk diplotype (112/121) reported for Mexican-Americans and Northern Europeans, were comparable between patients and controls.

Conclusions

CAPN10 UCSNP-19 variant, and the 111 haplotype contribute to the risk of T2D in Tunisian subjects; no significant associations between CAPN10 diplotypes and T2D were demonstrated for Tunisians.
Late tamoxifen in patients previously operated for breast cancer without postoperative tamoxifen: 5-year results of a single institution randomised study
Authors: Veronesi Andrea , Miolo GianMaria , Magri Maria D , Crivellari Diana , Scalone Simona , Bidoli Ettore , Lombardi Davide
Published: May 2010

Background

A population of breast cancer patients exists who, for various reasons, never received adjuvant post-operative tamoxifen (TAM). This study was aimed to evaluate the role of late TAM in these patients.

Methods

From 1997 to 2003, patients aged 35 to 75 years, operated more than 2 years previously for monolateral breast cancer without adjuvant TAM, with no signs of metastases and no contraindication to TAM were randomized to TAM 20 mg/day orally for 2 years or follow-up alone. Events were categorized as locoregional relapse, distant metastases, metachronous breast cancer, tumours other than breast cancer and death from any causes, whichever occurred first. The sample size (197 patients per arm, plus 10% allowance) was based on the assumption of a 30% decrease in the number of events occurring at a rate of 5% annually in the 10 years following randomization. Four hundred and thirty-three patients were randomized in the study (TAM 217, follow-up 216). Patients characteristics (TAM/follow-up) included: median age 55/55 years, median time from surgery 25/25 months (range, 25-288/25-294), in situ carcinoma 18/24, oestrogen receptor (ER) positive in 75/68, negative in 70/57, unknown in 72/91 patients. Previous adjuvant treatment included chemotherapy in 131/120 and an LHRH analogue in 11/13 patients.

Results

Thirty-six patients prematurely discontinued TAM after a median of 1 month, mostly because of subjective intolerance. Eighty-three events (TAM 39, follow-up 44) occurred: locoregional relapse in 10/8, distant metastases in 14/16, metachronous breast cancer in 4/10, other tumours in 11/10 patients. Less ER-positive secondary breast cancers occurred in the TAM treated patients than in follow-up patients (1 vs 10, p = 0.005). Event-free survival was similar in both groups of patients.

Conclusions

This 5-year analysis revealed significantly less metachronous ER-positive breast cancers in the TAM treated patients. No other statistically significant differences have emerged thus far.
Decreased transcription-coupled nucleotide excision repair capacity is associated with increased p53- and MLH1-independent apoptosis in response to cisplatin
Authors: Stubbert Lawton J , Smith Jennifer M , McKay Bruce C
Published: May 2010

Background

One of the most commonly used classes of anti-cancer drugs presently in clinical practice is the platinum-based drugs, including cisplatin. The efficacy of cisplatin therapy is often limited by the emergence of resistant tumours following treatment. Cisplatin resistance is multi-factorial but can be associated with increased DNA repair capacity, mutations in p53 or loss of DNA mismatch repair capacity.

Methods

RNA interference (RNAi) was used to reduce the transcription-coupled nucleotide excision repair (TC-NER) capacity of several prostate and colorectal carcinoma cell lines with specific defects in p53 and/or DNA mismatch repair. The effect of small inhibitory RNAs designed to target the CSB (Cockayne syndrome group B) transcript on TC-NER and the sensitivity of cells to cisplatin-induced apoptosis was determined.

Results

These prostate and colon cancer cell lines were initially TC-NER proficient and RNAi against CSB significantly reduced their DNA repair capacity. Decreased TC-NER capacity was associated with an increase in the sensitivity of tumour cells to cisplatin-induced apoptosis, even in p53 null and DNA mismatch repair-deficient cell lines.

Conclusion

The present work indicates that CSB and TC-NER play a prominent role in determining the sensitivity of tumour cells to cisplatin even in the absence of p53 and DNA mismatch repair. These results further suggest that CSB represents a potential target for cancer therapy that may be important to overcome resistance to cisplatin in the clinic.
Comparison of group counseling with individual counseling in the comprehension of informed consent: a randomized controlled trial
Authors: Sarkar Rajiv , Sowmyanarayanan Thuppal V , Samuel Prasanna , Singh Azara S , Bose Anuradha , Muliyil Jayaprakash , Kang Gagandeep
Published: May 2010

Background

Studies on different methods to supplement the traditional informed consent process have generated conflicting results. This study was designed to evaluate whether participants who received group counseling prior to administration of informed consent understood the key components of the study and the consent better than those who received individual counseling, based on the hypothesis that group counseling would foster discussion among potential participants and enhance their understanding of the informed consent.

Methods

Parents of children participating in a trial of nutritional supplementation were randomized to receive either group counseling or individual counseling prior to administration of the informed consent. To assess the participant's comprehension, a structured questionnaire was administered approximately 48-72 hours afterwards by interviewers who were blinded to the allocation group of the respondents.

Results

A total of 128 parents were recruited and follow up was established with 118 (90.2%) for the study. All respondents were aware of their child's participation in a research study and the details of sample collection. However, their understanding of study purpose, randomization and withdrawal was poor. There was no difference in comprehension of key elements of the informed consent between the intervention and control arm.

Conclusions

The results suggest that the group counseling might not influence the overall comprehension of the informed consent process. Further research is required to devise better ways of improving participants' understanding of randomization in clinical trials.

Trial Registration

Clinical Trial Registry - India (CTRI): CTRI/2009/091/000612