bluuebunny/crossref_metadata_2025 huggingface dataset:
Metaphysics and the Origins of Modern Science: Descartes and the Importance of Laws of Nature
Authors: Henry John
Published: June 2004

This paper draws attention to the crucial importance of a new kind of precisely defined law of nature in the Scientific Revolution. All explanations in the mechanical philosophy depend upon the interactions of moving material particles; the laws of nature stipulate precisely how these interact; therefore, such explanations rely on the laws of nature. While this is obvious, the radically innovatory nature of these laws is not fully acknowledged in the historical literature. Indeed, a number of scholars have tried to locate the origins of such laws in the medieval period. In the first part of this paper these claims are critically examined, and found at best to reveal important aspects of the background to the later idea, which could be drawn upon for legitimating purposes by the mechanical philosophers. The second part of the paper argues that the modern concept of laws of nature originates in René Descartes's work. It is shown that Descartes took his concept of laws of nature from the mathematical tradition, but recognized that he could not export it to the domain of physico-mathematics, to play a causal role, unless he could show that these laws were underwritten by God. It is argued that this is why, at an early stage of his philosophical development, Descartes had to turn to metaphysics.
Batak and Wealth: A Critical Study of Materialism in the Batak Churches in Indonesia
Authors: Sihombing Batara
Published: June 2004

In this article, Indonesian theologian Batara Sihombing analyses the place and function of wealth among the Toba Batak of Northern Sumatra, the people into whom he was born. After surveying the Batak kinship system and the Batak people in general, Sihombing reflects on the three highest ideals of the Batak people and provides a number of proverbs which point to the centrality of wealth and honor in Batak culture. In a third section of his essay, Sihombing reflects on the practices of auction, patronage and corruption among the Batak, and offers a strong critique of each in the light of Scripture and Christian principles.
A Pastoral Approach to Spirit Possession and Witchcraft Manifestations among the Fijian People
Authors: Hoare Frank
Published: June 2004

In this article Columban missionary, Frank Hoare, examines incidents of spirit possession and witchcraft accusations in a Catholic Fijian village. Spirit possession has a communal dimension in socio-centric cultures and the tensions of the community are played out in and through the messages of the spirit. Crises of evil are understood as essentially moral and connect the individual, community and spirit world and the community is energized to resolve the situation and ward off the evil. The recognition by a community of individuals who have effective spiritual power may surface alternative dormant ritual economics and politics based on a traditional cosmology that may be more or less articulated to the dominant cosmology. A foreign missionary should beware of ethnocentrism and the reductionism that dismisses the local idiom and traditional cosmology. Instead, crises of evil offer an opportunity for deeper dialogue between the Christian gospel and traditional beliefs and practice. Study of the history, social relations and culture of the community is time well spent and the missionary should rely on mature local Christians. A liturgy that takes community crises into account can provide a wider context of meaning for the concerns of the community.
Flt3 ligand enhances thymic-dependent and thymic-independent immune reconstitution
Authors: Fry Terry J. , Sinha Manoj , Milliron Matthew , Chu Yu-Waye , Kapoor Veena , Gress Ronald E. , Thomas Elaine , Mackall Crystal L.
Published: June 2004

Despite recent progress in our understanding of the biology of T-cell homeostasis, clinically available therapies to substantially improve immune reconstitution in patients sustaining T-cell depletion are lacking. T cells are regenerated via a dynamic interplay between thymopoiesis and thymic-independent homeostatic peripheral expansion (HPE). Using athymic mice subjected to T-cell depletion, we observed that HPE is critically dependent on dendritic cells (DCs) for presentation of antigen, raising the possibility that the availability of DCs might be limiting in vivo for HPE to occur efficiently. Indeed, flt3 ligand (flt3L) treatment of athymic mice subjected to T-cell depletion (without DC depletion) substantially enhanced HPE and improved immune competence. Following bone marrow transplantation (BMT) in athymic hosts, both dendritic cells and T cells were profoundly depleted and flt3L therapy restored DC numbers and enhanced HPE. In addition, thymus-bearing BMT recipients treated with flt3L regenerated increased numbers of thymic-dependent progeny with increased numbers of T-cell receptor excision circle (TREC)-positive T cells, indicating increased thymopoiesis. Therefore, flt3L is a potent immunorestorative agent that enhances both thymic-dependent and thymic-independent pathways of T-cell regeneration. (Blood. 2004;104:2794-2800)
Chlorogenic acid inhibits Bcr-Abl tyrosine kinase and triggers p38 mitogen-activated protein kinase–dependent apoptosis in chronic myelogenous leukemic cells
Authors: Bandyopadhyay Gautam , Biswas Tanusree , Roy Keshab C. , Mandal Swapan , Mandal Chhabinath , Pal Bikas C. , Bhattacharya Samir , Rakshit Srabanti , Bhattacharya Dilip K. , Chaudhuri Utpal , Konar Aditya , Bandyopadhyay Santu
Published: June 2004

We report that chlorogenic acid (Chl) induces apoptosis of several Bcr-Abl–positive chronic myelogenous leukemia (CML) cell lines and primary cells from CML patients in vitro and destroys Bcr-Abl–positive K562 cells in vivo. In contrast, this compound has no effect on the growth and viability of Bcr-Abl–negative lymphocytic and myeloid cell lines and primary CML cells. Sodium chlorogenate (NaChl) exhibits 2-fold higher efficiency in killing K562 cells compared with Chl. NaChl also induces growth inhibition of squamous cell carcinoma (HSC-2) and salivary gland tumor cells (HSG), although at 50-fold higher concentration. NaChl inhibits autophosphorylation of p210Bcr-Abl fusion protein rapidly. We demonstrate that p38 phosphorylation is increased in Bcr-Abl–positive cells after treatment with NaChl and closely paralleled the inhibition of Bcr-Abl phosphorylation. NaChl did not increase phosphorylation of p38 in Bcr-Abl–negative cells including HSC-2 and HSG that are responsive to this compound, indicating that p38 activation by NaChl is dependent on Bcr-Abl kinase inhibition. Inhibition of p38 activity by SB203580 significantly reduced NaChl-induced apoptosis of K562 cells, whereas activation of p38 by anisomycin augmented the apoptosis. These findings indicate that inhibition of Bcr-Abl kinase leading to activation of p38 mitogen-activated protein (MAP) kinase may play an important role in the anti-CML activity of Chl.
The tetraspanin superfamily member CD151 regulates outside-in integrin αIIbβ3 signaling and platelet function
Authors: Lau Lai-Man , Wee Janet L. , Wright Mark D. , Moseley Gregory W. , Hogarth P. Mark , Ashman Leonie K. , Jackson Denise E.
Published: June 2004

The tetraspanin family member CD151 forms complexes with integrins and regulates cell adhesion and migration. While CD151 is highly expressed in megakaryocytes and to a lesser extent in platelets, its physiologic role in platelets is unclear. In this study, we investigate the physical and functional importance of CD151 in murine platelets. Immunoprecipitation/Western blot studies reveal a constitutive physical association of CD151 with integrin αIIbβ3 complex under strong detergent conditions. Using CD151-deficient mice, we show that the platelets have impaired “outside-in” integrin αIIbβ3 signaling with defective platelet aggregation responses to protease-activated receptor 4 (PAR-4) agonist peptide, collagen, and adenosine diphosphate (ADP); impaired platelet spreading on fibrinogen; and delayed kinetics of clot retraction in vitro. This functional integrin αIIbβ3 defect could not be attributed to altered expression of integrin αIIbβ3. CD151–/– platelets displayed normal platelet alpha granule secretion, dense granule secretion, and static platelet adhesion. In addition, CD151–/– platelets displayed normal “inside-out” integrin αIIbβ3 signaling properties as demonstrated by normal agonist-induced binding of soluble fluorescein isothiocyanate (FITC)–fibrinogen, JON/A antibody binding, and increases in cytosolic-free calcium and inositol 1,4,5 triphosphate (IP3) levels. This study provides the first direct evidence that CD151 is essential for normal platelet function and that disruption of CD151 induced a moderate outside-in integrin αIIbβ3 signaling defect.
An association of candidate gene haplotypes and bleeding severity in von Willebrand disease (VWD) type 1 pedigrees
Authors: Kunicki Thomas J. , Federici Augusto B. , Salomon Daniel R. , Koziol James A. , Head Steven R. , Mondala Tony S. , Chismar Jeffrey D. , Baronciani Luciano , Canciani Maria Teresa , Peake Ian R.
Published: June 2004

von Willebrand disease (VWD) type 1 is difficult to diagnose because of bleeding variability and low heritability of von Willebrand factor (VWF) levels. We compared a bleeding severity score and bleeding times to candidate gene haplotypes within pedigrees of 14 index cases, using a covariance components model for multivariate traits (Mendel: QTL Association). These pedigrees included 13 affected and 40 unaffected relatives, as defined by plasma ristocetin cofactor (VWF:RCo) levels. The bleeding severity score was derived from a detailed history. Donors were genotyped using a primer extension method, and 9 candidate genes were selected for analysis. VWF:RCo levels had the strongest influence on bleeding severity score and bleeding time. ITGA2 haplotype 2 (807C) and ITGA2B haplotype 1 (Ile843) were each associated with increased bleeding severity scores (P < .01 and P < .01, respectively). GP6 haplotype b (Pro219) was also associated with increased scores (P = .03) after adjustment for donor age. No association was observed with 6 other candidate genes, GP1BA, ITGB3, VWF, FGB, IL6, or TXA2R. Increased plasma VWF:Ag levels were associated with VWF haplotype 1 (–1793G; P = .02). These results establish that genetic differences in the adhesion receptor subunits α2, αIIb, and GPVI can influence the phenotype of VWD type 1.
Cyclin D1 activation in B-cell malignancy: association with changes in histone acetylation, DNA methylation, and RNA polymerase II binding to both promoter and distal sequences
Authors: Liu Hui , Wang Jin , Epner Elliot M.
Published: June 2004

Cyclin D1 expression is deregulated by chromosome translocation in mantle cell lymphoma and a subset of multiple myeloma. The molecular mechanisms involved in long-distance gene deregulation remain obscure, although changes in acetylated histones and methylated CpG dinucleotides may be important. The patterns of DNA methylation and histone acetylation were determined at the cyclin D1 locus on chromosome 11q13 in B-cell malignancies. The cyclin D1 promoter was hypomethylated and hyperacetylated in expressing cell lines and patient samples, and methylated and hypoacetylated in nonexpressing cell lines. Domains of hyperacetylated histones and hypomethylated DNA extended over 120 kb upstream of the cyclin D1 gene. Interestingly, hypomethylated DNA and hyperacetylated histones were also located at the cyclin D1 promoter but not the upstream major translocation cluster region in cyclin D1-nonexpressing, nontumorigenic B and T cells. RNA polymerase II binding was demonstrated both at the cyclin D1 promoter and 3′ immunoglobulin heavy-chain regulatory regions only in malignant B-cell lines with deregulated cyclin D1 expression. Our results suggest a model where RNA polymerase II bound at IgH regulatory sequences can activate the cyclin D1 promoter by either long-range polymerase transfer or tracking.
From the bench to the bedside: ways to improve rituximab efficacy
Authors: Cartron Guillaume , Watier Hervé , Golay Josée , Solal-Celigny Philippe
Published: June 2004

Rituximab (MabThera, Rituxan) is a chimeric IgG1 monoclonal antibody that specifically targets the CD20 surface antigen expressed on normal and neoplastic B-lymphoid cells. Rituximab is currently used in the treatment of both follicular and aggressive B-cell non-Hodgkin lymphomas. Despite its demonstrated clinical effectiveness, its in vivo mechanisms of action remain unknown and could differ by subtype of lymphoma. Rituximab has been shown to induce apoptosis, complement-mediated lysis, and antibody-dependent cellular cytotoxicity in vitro, and some evidence points toward an involvement of these mechanisms in vivo. Rituximab also has a delayed therapeutic effect as well as a potential “vaccinal” effect. Here, we review the current understanding of the mechanism of action of rituximab and discuss approaches that could increase its clinical activity. A better understanding of how rituximab acts in vivo should make it possible to develop new and more effective therapeutic strategies. (Blood. 2004;104:2635-2642)
In vitro and in vivo evaluation of a new active heat moisture exchanger
Authors: Chiumello Davide , Pelosi Paolo , Park Gilbert , Candiani Andrea , Bottino Nicola , Storelli Ezio , Severgnini Paolo , D'Onofrio Dunia , Gattinoni Luciano , Chiaranda Massimo
Published: June 2004

Introduction

In order to improve the efficiency of heat moisture exchangers (HMEs), new hybrid humidifiers (active HMEs) that add water and heat to HMEs have been developed. In this study we evaluated the efficiency, both in vitro and in vivo, of a new active HME (the Performer; StarMed, Mirandola, Italy) as compared with that of existing HMEs (Hygroster and Hygrobac; Mallinckrodt, Mirandola, Italy).

Methods

We tested the efficiency by measuring the temperature and absolute humidity (AH) in vitro using a test lung ventilated at three levels of minute ventilation (5, 10 and 15 l/min) and at two tidal volumes (0.5 and 1 l), and in vivo in 42 patients with acute lung injury (arterial oxygen tension/fractional inspired oxygen ratio 283 ± 72 mmHg). We also evaluated the efficiency in vivo after 12 hours.

Results

In vitro, passive Performer and Hygrobac had higher airway temperature and AH (29.2 ± 0.7°C and 29.2 ± 0.5°C, [P < 0.05]; AH: 28.9 ± 1.6 mgH2O/l and 28.1 ± 0.8 mgH2O/l, [P < 0.05]) than did Hygroster (airway temperature: 28.1 ± 0.3°C [P < 0.05]; AH: 27 ± 1.2 mgH2O/l [P < 0.05]). Both devices suffered a loss of efficiency at the highest minute ventilation and tidal volume, and at the lowest minute ventilation. Active Performer had higher airway temperature and AH (31.9 ± 0.3°C and 34.3 ± 0.6 mgH2O/l; [P < 0.05]) than did Hygrobac and Hygroster, and was not influenced by minute ventilation or tidal volume. In vivo, the efficiency of passive Performer was similar to that of Hygrobac but better than Hygroster, whereas Active Performer was better than both. The active Performer exhibited good efficiency when used for up to 12 hours in vivo.

Conclusion

This study showed that active Performer may provide adequate conditioning of inspired gases, both as a passive and as an active device.
Advance statements in old age psychiatry
Authors: Williams Lisa , Rigby John
Published: June 2004

Advance statements, originally designed for situations of terminal illness, are becoming increasingly relevant in psychiatry. This article examines their background, rationale and use in old age psychiatry. The difference between advance statements and advance directives is highlighted, and different roles are suggested for each. We discuss the advantages, and the possible limitations, of advance statements, including the unique issue of the Mental Health Act 1983.
Psychodynamic lessons in risk assessment and management1
Authors: Doctor Ronald
Published: June 2004

Risk assessment is notoriously difficult and offers a particular challenge to mental health workers. There are two main conceptual approaches to the consideration of risk assessment and management: the actuarial and the clinical methods. I use case material to examine the clinical, and in particular the psychodynamic, approach in order to illustrate that violent behaviour has meaning. Understanding its meaning and learning from it can substantially reduce the risks of further violent behaviour. Managing risk with the aid of psychodynamic psychotherapy enables the patient to become aware of his or her own mind and its function. Although this process may initially be distressing to the patient, in the longer term the aim is to enable the development of a healthier, more stable internal reality. This should, in turn, reduce the risk of dangerous acting out.
Violent figures, risky stories
Authors: Witteman Cilia
Published: June 2004

Doctor (2004, this issue) raises the important question of how to assess and manage violent behaviour. He claims that actuarial models of risk assessment based on epidemiology have failed, and that indeed the most reliable risk assessment is one based on clinical methods. He proposes that psychodynamic psychotherapy be used for this purpose, to uncover the meaning of the violent behaviour. Such therapy will, Doctor argues, help the violent person be aware of and understand the function of their behaviour. This understanding in turn will reduce the need in the patient to act out violently.
The doctor–nurse relationship
Authors: Fagin Leonard , Garelick Antony
Published: June 2004

In this article on getting on with colleagues in the workplace we explore how the nurse–doctor relationship in psychiatry has evolved and discuss its current status in both the in-patient ward and community mental health teams. In particular, we outline the changed roles and expanding responsibilities of nurses in the UK today. We suggest ways in which doctors can improve the relationship and give areas of possible future collaboration between doctors and nurses.
Decision-making about children's mental health care: ethical challenges
Authors: Paul Moli
Published: June 2004

Children and young people are usually referred to specialist child and adolescent mental health services (CAMHS) by adults because of concerns raised by other adults. Most CAMHS consider them from a developmental perspective and as individuals in the context of their families and other relationships/systems. In this article I discuss ethical and legal challenges posed by making decisions with and about children and young people within CAMHS, with particular reference to duty of care; the rights of minors' and parents'; capacity and consent; and disagreement between decision-makers. It is important to involve children and young people in decision-making, and I suggest ways of acheiving this.
Boundary violation and sexual exploitation in psychiatry and psychotherapy: a review
Authors: Sarkar Sameer P.
Published: June 2004

In psychiatric and psychotherapeutic practice, ‘boundaries' delineate the personal and the professional roles and the differences that should characterise the interpersonal encounters between the patient/client and the professional. Boundaries are essential to keep both parties safe. The author outlines the various types of boundary violation that can arise in clinical practice, their consequences (both clinical and legal), how professionals can avoid them and how health care institutions might respond, should they occur. He concentrates on sexual boundary violations, because these have been the subject of most empirical study.
Direct-to-consumer advertising of psychotropics
Authors: Gilbody Simon , Wilson Paul , Watt Ian
Published: June 2004

‘Advertising may be described as the science of arresting the human intelligence long enough to get money from it’ (Leacock, 1924).

Until recently, the main targets of pharmaceutical advertisements were clinicians and the major vehicles for ‘educational promotion’ were advertisements in academic and professional journals and visits and gifts from pharmaceutical representatives. All is now changed and patients have become legitimate targets of advertising. Direct-to-consumer advertising represents a new and expanding area of pharmaceutical promotion that harnesses the mass media.
Cumulative incidence of mental disorders among offspring of mothers with psychotic disorder
Authors: Niemi Laura T. , Suvisaari Jaana M. , Haukka Jari K. , Wrede Gunnel , Lönnqvist Jouko K.
Published: June 2004

Background

The Helsinki High-Risk Study follows up all women born between 1916 and 1948 and treated for schizophrenia-spectrum disorders in psychiatric hospitals in Helsinki, their offspring born between 1960 and 1964, and controls.

Aims

To determine the cumulative incidence of adulthood Axis I disorders among offspring.

Method

Using all hospital and out-patient treatment records we rediagnosed parents and offspring according to DSM– IV-TR criteria. Offspring were grouped by mother's diagnosis (schizophrenia n = 104, schizoaffective disorder n=20, other schizophrenia-spectrum disorder n=30, and affective disorder n=25) and compared with a control group (n = 176). The cumulative incidences of Axis I disorders among offspring were calculated.

Results

The cumulative incidences of any psychotic disorder were 13.5%, 10.0%, 10.0%, 4.0% and 1.1% among offspring of mothers with schizophrenia, schizo-affective disorder, other schizophrenia-spectrum disorders, affective disorders and controls, respectively. The corresponding figures for schizophrenia were 6.7%, 5.0%, 6.7%, 0% and 0.6%, and for any mental disorder 23.1%, 20.0%, 20.0%, 12.0% and 6.9%.

Conclusions

Offspring of mothers with a psychotic disorder have heightened risk of developing a wide range of severe mental disorders.
Explanatory models of illness in schizophrenia: Comparison of four ethnic groups
Authors: McCabe Rosemarie , Priebe Stefan
Published: June 2004

Background

Explanatory models of illness may differ between ethnic groups and influence treatment satisfaction and compliance.

Aims

To compare explanatory models among people with schizophrenia from four cultural backgrounds and explore their relationship with clinical and psychological characteristics.

Method

Explanatory models, insight, treatment compliance, health locus of control, quality of life, treatment satisfaction, therapeutic relationships and symptomatology were assessed in UK Whites and Bangladeshis, African-Caribbeans and West Africans.

Results

When biological and supernatural causes of illness were compared, Whites cited biological causes more frequently than the three non-White groups, who cited supernatural causes more frequently. When biological and social causes were compared, Whites cited biological causes more frequently than African-Caribbeans and Bangladeshis, who cited social causes more frequently. A biological explanatory model was related to enhanced treatment satisfaction and therapeutic relationships but not treatment compliance.

Conclusions

Explanatory models of illness contribute to patient satisfaction with treatment and relationships with clinicians.
Clinical efficacy of computerised cognitive-behavioural therapy for anxiety and depression in primary care: Randomised controlled trial
Authors: Proudfoot Judith , Ryden Clash , Everitt Brian , Shapiro David A. , Goldberg David , Mann Anthony , Tylee Andre , Marks Isaac , Gray Jeffrey A.
Published: June 2004

Background

Preliminary results have demonstrated the clinical efficacy of computerised cognitive-behavioural therapy (CBT) in the treatment of anxiety and depression in primary care.

Aims

To determine, in an expanded sample, the dependence of the efficacy of this therapy upon clinical and demographic variables.

Method

A sample of 274 patients with anxiety and/or depression were randomly allocated to receive, with or without medication, computerised CBTor treatment as usual, with follow-up assessment at 6 months.

Results

The computerised therapy improved depression, negative attributional style, work and social adjustment, without interaction with drug treatment, duration of preexisting illness or severity of existing illness. For anxiety and positive attributional style, treatment interacted with severity such that computerised therapy did better than usual treatment for more disturbed patients. Computerised therapy also led to greater satisfaction with treatment.

Conclusions

Computer-delivered CBT is a widely applicable treatment for anxiety and/or depression in general practice.
Repetition of deliberate self-harm and subsequent suicide risk: Long-term follow-up study of 11 583 patients
Authors: Zahl Daniel Louis , Hawton Keith
Published: June 2004

Background

Repetition of deliberate self-harm (DSH) is a risk factor for suicide. Little information is available on the risk for specific groups of people who deliberately harm themselves repeatedly.

Aims

To investigate the long-term risk of suicide associated with repetition of DSH by gender, age and frequency of repetition.

Method

A mortality follow-up study to the year 2000 was conducted on 11583 people who presented to the general hospital in Oxford between 1978 and 1997. Repetition of DSH was determined from reported episodes prior to the index episode and episodes presenting to the same hospital during the follow-up period. Deaths were identified through national registers.

Results

Thirty-nine percent of patients repeated the DSH. They were at greater relative risk of suicide than the single-episode DSH group (2.24; 95% CI 1.77–2.84). The relative risk of suicide in the repeated DSH group compared with the single-episode DSH group was greater in females (3.5; 95% C11.3–2.4) than males (1.8; 95% C1 2.3–5.3) and was inversely related to age (up to 54 years). Suicide risk increased further with multiple repeat episodes of DSH in females.

Conclusions

Repetition of DSH is associated with an increased risk of suicide in males and females. Repetition may be a better indicator of risk in females, especially young females.
Nucleolar localization and mobility analysis of the NF-κB repressing factor NRF
Authors: Niedick Ina , Froese Natali , Oumard André , Mueller Peter P. , Nourbakhsh Mahtab , Hauser Hansjörg , Köster Mario
Published: June 2004

NF-κB plays a central role in mediating pathogen and cytokine-stimulated gene transcription. NF-κB repressing factor (NRF) has been shown to interact with specific negative regulatory DNA elements (NRE) to mediate transcriptional repression by inhibition of the NF-κB activity at certain promoters. mRNA ablation experiments demonstrated that the trans-acting NRF protein is involved in constitutive but not post-stimulated silencing of IFN-β, IL-8 and iNOS genes by binding to cis-acting NRE elements in their promoters.

We have examined the subcellular localization and mobility of the NRF protein. Since neither tagging nor overexpression perturbs NRF localization the GFP-tagged protein was used for detailed localization and mobility studies. Owing to an N-terminal nuclear localization sequence, all NRF fragments that contain this signal show a constitutive nuclear accumulation. C-terminal NRF fragments also localize to the nucleus although no canonical NLS motifs were detected. Full-length NRF is highly enriched in nucleoli and only a small fraction of NRF is found in the nucleoplasm and cytoplasm. This relationship was found to be independent of the protein expression rate. FRAP analysis proved to be a sensitive method to determine protein mobility and made it possible to differentiate between the NRF protein fragments. Nucleolar localization correlated inversely with mobility. The data demonstrate that a series of neighboring fragments in a large central domain of the protein contribute to the strong nucleolar affinity. These properties were not altered by viral infection or LPS treatment. Several sequence motifs for RNA binding were predicted by computer-mediated databank searches. We found that NRF binds to double stranded RNA (dsRNA). This property mapped to several NRF fragments which correlate with the nucleolar affinity domain. Since treatment with actinomycin D releases NRF from nucleoli the identified RNA binding motifs might act as nucleolar localization signals.
Annexin 2 is a phosphatidylinositol (4,5)-bisphosphate binding protein recruited to actin assembly sites at cellular membranes
Authors: Rescher Ursula , Ruhe Daniela , Ludwig Carsten , Zobiack Nicole , Gerke Volker
Published: June 2004

Annexin 2 is a Ca2+-regulated membrane protein and an F-actin-binding protein enriched at actin assembly sites both, on the plasma membrane and on endosomal vesicles. Here, we identify annexin 2 as a phosphatidylinositol (4,5)-bisphosphate (PtdIns(4,5)P2)-interacting protein, thereby explaining this specific membrane association. Using the pleckstrin-homology (PH) domain of phospholipase Cδ1 fused to yellow fluorescent protein as a marker for PtdIns(4,5)P2, we show that annexin 2 and its ligand p11 (S100A10) are targeted to sites of PtdIns(4,5)P2 enrichment where F-actin accumulates. At the plasma membrane, adhesion of pedestal-forming enteropathogenic Escherichia coli induces a recruitment of 1-phosphatidylinositol-4-phosphate 5-kinase (PtdIns4P 5-kinase) and an enrichment of PtdIns(4,5)P2 and annexin 2-p11 at sites of bacterial adhesion. Induction of PtdIns(4,5)P2-enriched ruffles and PtdIns(4,5)P2-positive, actin-coated vacuoles by Arf6-mediated activation of PtdIns4P 5-kinase also leads to a concomitant accumulation of the annexin 2-p11 complex and the PH domain. Binding studies with immobilized phosphoinositides and phosphoinositide-containing liposomes reveal that the purified annexin 2-p11 complex directly and specifically binds to PtdIns(4,5)P2 with an affinity comparable to that of the PH domain of phospholipase Cδ1. Experiments using individual subunits identify annexin 2 as the PtdIns(4,5)P2-binding entity. Thus, the direct interaction of annexin 2 with PtdIns(4,5)P2 is a means of specifically recruiting the annexin 2-p11 complex to sites of membrane-associated actin assembly.
Caldesmon mutant defective in Ca2+-calmodulin binding interferes with assembly of stress fibers and affects cell morphology, growth and motility
Authors: Li Yan , Lin Jenny L. C. , Reiter Rebecca S. , Daniels Karla , Soll David R. , Lin Jim J. C.
Published: June 2004

Despite intensive in vitro studies, little is known about the regulation of caldesmon (CaD) by Ca2+-calmodulin (Ca2+-CaM) in vivo. To investigate this regulation, a mutant was generated of the C-terminal fragment of human fibroblast CaD, termed CaD39-AB, in which two crucial tryptophan residues involved in Ca2+-CaM binding were each replaced with alanine. The mutation abolished most CaD39-AB binding to Ca2+-CaM in vitro but had little effect on in vitro binding to actin filaments and the ability to inhibit actin/tropomyosin-activated heavy meromyosin ATPase. To study the functional consequences of these mutations in vivo, we transfected an expression plasmid carrying CaD39-AB cDNA into Chinese hamster ovary (CHO) cells and isolated several clones expressing various amounts of CaD39-AB. Immunofluorescence microscopy revealed that mutant CaD39-AB was distributed diffusely throughout the cytoplasm but also concentrated at membrane ruffle regions. Stable expression of CaD39-AB in CHO cells disrupted assembly of stress fibers and focal adhesions, altered cell morphology, and slowed cell cycle progression. Moreover, CaD39-AB-expressing cells exhibited motility defects in a wound-healing assay, in both velocity and the persistence of translocation, suggesting a role for CaD regulation by Ca2+-CaM in cell migration. Together, these results demonstrate that CaD plays a crucial role in mediating the effects of Ca2+-CaM on the dynamics of the actin cytoskeleton during cell migration.
Snake presynaptic neurotoxins with phospholipase A2 activity induce punctate swellings of neurites and exocytosis of synaptic vesicles
Authors: Rigoni Michela , Schiavo Giampietro , Weston Anne E. , Caccin Paola , Allegrini Federica , Pennuto Maria , Valtorta Flavia , Montecucco Cesare , Rossetto Ornella
Published: June 2004

The mechanisms of action of four snake presynaptic phospholipase A2 neurotoxins were investigated in cultured neurons isolated from various parts of the rat brain. Strikingly, physiological concentrations of notexin, β-bungarotoxin, taipoxin or textilotoxin induced a dose-dependent formation of discrete bulges at various sites of neuronal projections. Neuronal bulging was paralleled by the redistribution of the two synaptic vesicle markers synaptophysin I (SypI) and vesicle-attached membrane protein 2 (VAMP2) to the bulges, and by the exposure of the luminal domain of synaptotagmin on the cell surface. These neurotoxins induced glutamate release from cultured neurons similarly to the known evoked release of acetylcholine from neuromuscular junctions. In addition, partial fragmentation of F-actin and neurofilaments was observed in neurons, but not in astrocytes. These findings indicate that these snake presynaptic neurotoxins act with by same mechanism and that the observed phenotype results from the fusion of synaptic vesicles with the plasma membrane not balanced by an adequate membrane retrieval. These changes closely resemble those occurring at neuromuscular junctions of intoxicated animals and fully qualify these primary neuronal cultures as pertinent models for studying the molecular mode of action of these neurotoxins.
CD133, a novel marker for human prostatic epithelial stem cells
Authors: Richardson Gavin D. , Robson Craig N. , Lang Shona H. , Neal David E. , Maitland Norman J. , Collins Anne T.
Published: June 2004

Stem cells are clonogenic cells with self-renewal and differentiation properties, which may represent a major target for genetic damage leading to prostate cancer and benign prostatic hyperplasia. Stem cells remain poorly characterised because of the absence of specific molecular markers that permit us to distinguish them from their progeny, the transit amplifying cells, which have a more restricted proliferative potential. Human CD133 antigen, also known as AC133, was recently identified as a haematopoietic stem cell marker. Here we show that a small population (approximately 1%) of human prostate basal cells express the cell surface marker CD133 and are restricted to the α2β1hi population, previously shown to be a marker of stem cells in prostate epithelia (Collins, A. T., Habib, F. K., Maitland, N. J. and Neal, D. E. (2001). J. Cell Sci. 114, 3865-3872). α2β1hi/CD133+ cells exhibit two important attributes of epithelial stem cells: they possess a high in vitro proliferative potential and can reconstitute prostatic-like acini in immunocompromised male nude mice.
Sister-chromatid cohesion mediated by the alternative RF-CCtf18/Dcc1/Ctf8, the helicase Chl1 and the polymerase-α-associated protein Ctf4 is essential for chromatid disjunction during meiosis II
Authors: Petronczki Mark , Chwalla Barbara , Siomos Maria F. , Yokobayashi Shihori , Helmhart Wolfgang , Deutschbauer Adam M. , Davis Ronald W. , Watanabe Yoshinori , Nasmyth Kim
Published: June 2004

Cohesion between sister chromatids mediated by a multisubunit complex called cohesin is established during DNA replication and is essential for the orderly segregation of chromatids during anaphase. In budding yeast, a specialized replication factor C called RF-CCtf18/Dcc1/Ctf8 and the DNA-polymerase-α-associated protein Ctf4 are required to maintain sister-chromatid cohesion in cells arrested for long periods in mitosis. We show here that CTF8, CTF4 and a helicase encoded by CHL1 are required for efficient sister chromatid cohesion in unperturbed mitotic cells, and provide evidence that Chl1 functions during S-phase. We also show that, in contrast to mitosis, RF-CCtf18/Dcc1/Cft8, Ctf4 and Chl1 are essential for chromosome segregation during meiosis and for the viability of meiotic products. Our finding that cells deleted for CTF8, CTF4 or CHL1 undergo massive meiosis II non-disjunction suggests that the second meiotic division is particularly sensitive to cohesion defects. Using a functional as well as a cytological assay, we demonstrate that CTF8, CHL1 and CTF4 are essential for cohesion between sister centromeres during meiosis but dispensable for cohesin's association with centromeric DNA. Our finding that mutants in fission yeast ctf18 and dcc1 have similar defects suggests that the involvement of the alternative RF-CCtf18/Dcc1/Ctf8 complex in sister chromatid cohesion might be highly conserved.
Differentiation of Acute Myeloid Leukemia from B- and T-Lineage Acute Lymphoid Leukemias by Real-Time Quantitative Reverse Transcription-PCR of Lineage Marker mRNAs
Authors: Saussoy Pascale , Vaerman Jean-Luc , Straetmans Nicole , Deneys Véronique , Cornu Guy , Ferrant Augustin , Latinne Dominique
Published: June 2004

Background: Flow cytometry of lineage markers is useful in the classification of leukemias. Our aim was to assess whether the study of lineage genes at the RNA level would enable differentiation of acute myeloid leukemias (AMLs) from B-and T-lineage acute lymphoid leukemias (ALLs).

Methods: We measured mRNA of four lineage markers [CD19, CD79a, CD3e, and myeloperoxidase (MPO)] by reverse transcription followed by real-time quantitative (RTQ)-PCR. We investigated 72 acute leukemias (40 AMLs with 23–93% blast cells plus 27 B-lineage ALLs and 5 T-lineage ALLs) defined by morphologic criteria at diagnosis. RTQ-PCR analysis was performed on bone marrow without cell sorting. The expression of each gene was calculated as the difference in the threshold cycle [ΔCT; CT value of target gene minus CT value of housekeeping gene (Abelson)].

Results: Three patterns of expression were detected. In the first, CD19, CD79a, and MPO mRNAs were less abundant than CD3e. In the second pattern, MPO mRNA was more abundant than the other three mRNAs. In the third, CD19 or CD79a was more highly expressed than CD3e and MPO. The three patterns corresponded to T-ALL, AML, and B-ALL, respectively. The use of cutoffs to establish qualitatively the pattern of coexpression of the four lineage markers provided the same information as the comparison among the four ΔCT values. Prospective use of the scoring system correctly classified each of 13 additional cases (8 AML, 4 B-lineage ALL, and 1 T-lineage ALL).

Conclusion: Study of lineage markers at diagnosis by RTQ-PCR allows differentiation of AML from B-ALL or T-ALL without cell sorting, even when the bone marrow contains few blast cells.
Genotyping of Single-Nucleotide Polymorphisms by High-Resolution Melting of Small Amplicons
Authors: Liew Michael , Pryor Robert , Palais Robert , Meadows Cindy , Erali Maria , Lyon Elaine , Wittwer Carl
Published: June 2004

Background: High-resolution melting of PCR amplicons with the DNA dye LCGreen™ I was recently introduced as a homogeneous, closed-tube method of genotyping that does not require probes or real-time PCR. We adapted this system to genotype single-nucleotide polymorphisms (SNPs) after rapid-cycle PCR (12 min) of small amplicons (≤50 bp).

Methods: Engineered plasmids were used to study all possible SNP base changes. In addition, clinical protocols for factor V (Leiden) 1691G>A, prothrombin 20210G>A, methylenetetrahydrofolate reductase (MTHFR) 1298A>C, hemochromatosis (HFE) 187C>G, and β-globin (hemoglobin S) 17A>T were developed. LCGreen I was included in the reaction mixture before PCR, and high-resolution melting was obtained within 2 min after amplification.

Results: In all cases, heterozygotes were easily identified because heteroduplexes altered the shape of the melting curves. Approximately 84% of human SNPs involve a base exchange between A::T and G::C base pairs, and the homozygotes are easily genotyped by melting temperatures (Tms) that differ by 0.8–1.4 °C. However, in ∼16% of SNPs, the bases only switch strands and preserve the base pair, producing very small Tm differences between homozygotes (<0.4 °C). Although most of these cases can be genotyped by Tm, one-fourth (4% of total SNPs) show nearest-neighbor symmetry, and, as predicted, the homozygotes cannot be resolved from each other. In these cases, adding 15% of a known homozygous genotype to unknown samples allows melting curve separation of all three genotypes. This approach was used for the HFE 187C>G protocol, but, as predicted from the sequence changes, was not needed for the other four clinical protocols.

Conclusions: SNP genotyping by high-resolution melting analysis is simple, rapid, and inexpensive, requiring only PCR, a DNA dye, and melting instrumentation. The method is closed-tube, performed without probes or real-time PCR, and can be completed in less than 2 min after completion of PCR.
Intrinsic Resistance of Oligodendrocytes to Prion Infection
Authors: Prinz Marco , Montrasio Fabio , Furukawa Hisako , van der Haar Marjan E. , Schwarz Petra , Rülicke Thomas , Giger Olivier T. , Häusler Karl-Georg , Perez Daniel , Glatzel Markus , Aguzzi Adriano
Published: June 2004

Within the CNS, the normal form of cellular prion protein (PrPC) is expressed on neurons, oligodendrocytes, and astrocytes. The contribution of these cell types to prion replication and pathogenesis is unclear. To assess the role of oligodendrocytes, we expressed PrPCunder the control of the myelin basic protein (MBP) promoter in mice lacking endogenous PrPC. PrPCwas detected in oligodendrocytes and Schwann cells but not in neurons and astrocytes. MBP-PrP mice never developed scrapie after intracerebral, intraperitoneal, or intraocular challenge with scrapie prions. Transgenic brains did not contain protease-resistant prion protein and did not transmit scrapie when inoculated into PrPC-overexpressing indicator mice. To investigate whether prion spread within the CNS depends on oligodendrocytic PrPC, we implanted PrPC-overexpressing neuroectodermal grafts into MBP-PrP brains. After intraocular prion inoculation, none of the grafts showed spongiform encephalopathy or prion infectivity. Hence oligodendrocytes do not support cell-autonomous prion replication, establishment of subclinical disease, and neural spread of prions. Prion resistance sets oligodendrocytes aside from both neurons and astrocytes.
The γ2 Subunit of GABAAReceptors Is a Substrate for Palmitoylation by GODZ
Authors: Keller Cheryl A. , Yuan Xu , Panzanelli Patrizia , Martin Michelle L. , Alldred Melissa , Sassoè-Pognetto Marco , Lüscher Bernhard
Published: June 2004

The neurotransmitter GABA activates heteropentameric GABAAreceptors, which are composed mostly of α, β, and γ2 subunits. Regulated membrane trafficking and subcellular targeting of GABAAreceptors is important for determining the efficacy of GABAergic inhibitory function. Of special interest is the γ2 subunit, which is mostly dispensable for assembly and membrane insertion of functional receptors but essential for accumulation of GABAAreceptors at synapses. In a search for novel receptor trafficking proteins, we have used the SOS-recruitment system and isolated a Golgi-specific DHHC zinc finger protein (GODZ) as a novel γ2 subunit-interacting protein. GODZ is a member of the superfamily of DHHC cysteine-rich domain (DHHC-CRD) polytopic membrane proteins shown recently in yeast to represent palmitoyltransferases. GODZ mRNA is found in many tissues; however, in brain the protein is detected in neurons only and highly concentrated and asymmetrically distributed in the Golgi complex. GODZ interacts with a cysteine-rich 14-amino acid domain conserved specifically in the large cytoplasmic loop of γ1-3 subunits but not in other GABAAreceptor subunits. Coexpression of GODZ and GABAAreceptors in heterologous cells results in palmitoylation of the γ2 subunit in a cytoplasmic loop domain-dependent manner. Neuronal GABAAreceptors are similarly palmitoylated. Thus, GODZ-mediated palmitoylation represents a novel posttranslational modification that is selective forγ subunit-containing GABAAreceptor subtypes, a mechanism that is likely to be important for regulated trafficking of these receptors in the secretory pathway.
Intrinsic and Circuit Properties Favor Coincidence Detection for Decoding Oscillatory Input
Authors: Perez-Orive Javier , Bazhenov Maxim , Laurent Gilles
Published: June 2004

In the insect olfactory system the antennal lobe generates oscillatory synchronization of its output as a framework for coincidence detection by its target, the mushroom body (MB). The intrinsic neurons of the MB (Kenyon cells, KCs) are thus a good model system in which to investigate the functional relevance of oscillations and neural synchronization. We combine electrophysiological and modeling approaches to examine how intrinsic and circuit properties might contribute to the preference of KCs for coincident input and how their decoding of olfactory information is affected by the absence of oscillatory synchronization in their input. We show that voltage-dependent subthreshold properties of KCs bring about a supralinear summation of their inputs, favoring responses to coincident EPSPs. Abolishing oscillatory synchronization weakens the preference of KCs for coincident input and causes a large reduction in their odor specificity. Finally, we find that a decoding strategy that is based on coincidence detection enhances both noise tolerance and input discriminability by KCs.
YB-1 and CTCF Differentially Regulate the 5-HTT Polymorphic Intron 2 Enhancer Which Predisposes to a Variety of Neurological Disorders
Authors: Klenova Elena , Scott Alison C. , Roberts Julian , Shamsuddin Shaharum , Lovejoy Elizabeth A. , Bergmann Stephan , Bubb Vivien J. , Royer Hans-Dieter , Quinn John P.
Published: June 2004

The serotonin transporter (5-HTT) gene contains a variable number tandem repeat (VNTR) domain within intron 2 that is often associated with a number of neurological conditions, including affective disorders. The implications of this polymorphism are not yet understood, however, we have previously demonstrated that the 5-HTT VNTR is a transcriptional regulatory domain, and the allelic variation supports differential reporter gene expressionin vivoandin vitro. The aim of this study was to identify transcription factors responsible for the regulation of this VNTR. Using a yeast one-hybrid screen, we found the transcription factor Y box binding protein 1 (YB-1) interacts with the 5-HTT VNTR. Consistent with this, we demonstrate in a reporter gene assay that the polymorphic VNTR domains differentially respond to exogenous YB-1 and that YB-1 will bind to the VNTRin vitroin a sequence-specific manner. Interestingly, the transcription factor CCTC-binding factor (CTCF), previously shown to interact with YB-1, interferes with the ability of the VNTR to support YB-1-directed reporter gene expression. In addition, CTCF blocks the binding of YB-1 to its DNA recognition sequencesin vitro, thus providing a possible mechanism of regulation of YB-1 activation of the VNTR by CTCF. Therefore, we have identified YB-1 and CTCF as transcription factors responsible, at least in part, for modulation of VNTR function as a transcriptional regulatory domain. Our data suggest a novel mechanism that explains, in part, the ability of the distinct VNTR copy numbers to support differential reporter gene expression based on YB-1 binding sites.