bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of an evidence‐based answering service on GPs and their patients: a pilot study
Authors: Verhoeven Anita A. H. , Schuling Jan 
Published: August 2004

AbstractObjectives:  For general practitioners (GPs), an important obstacle to practising evidence‐based medicine is lack of time. An evidence‐based answering service was developed that took over searching and appraisal of medical evidence from the GPs. GPs sent in questions, and the informationist formulated the answers. Our objectives were to find out if such an evidence‐based answering service was feasible, including assessing the effect of the answers on GPs and their patients, as reported by the GPs.Methods:  After attending a workshop on building well‐formulated questions from daily practice, the GPs sent in questions to the informationist. The literature was searched, the relevant information was appraised, and the answers to the questions were formulated. With a questionnaire, the effect of the answers on the GP and the patient was assessed, as well as the perceived barriers to implementing the answers.Results:  From 26 GPs, 61 questions were received. For 12% of questions, information was found with the highest evidence level, while for 36%, no information was found. However, for 89% of the questions for which no information could be found, the answer ‘no information found’ did have an effect on the GP concerned. In total, 81% of all the answers had an effect on the GP, and, according to the GP, 52% had an effect on the patient. Few barriers to implementing the evidence were perceived. Most of the answers were found in Pubmed/Medline, the Cochrane Library and Embase.Conclusions:  This study indicates that an evidence‐based answering service can have an impact on GPs and their patients. Librarians can provide an evidence‐based answering service for GPs and their patients. The evidence‐based answering service for GPs in this study had an impact on 81% of the GPs and on 52% of their patients. Although for one‐third of the questions no evidence‐based answer was found, this message in itself had an impact on 89% of the GPs. An informationist as mediator between medical information and doctors can save doctors’ time.
Clathrin-Dependent Targeting of Receptors to the Flagellar Pocket of Procyclic-Form
Trypanosoma brucei
Authors: Hung Chien-Hui , Qiao Xugang , Lee Pei-Tseng , Lee Mary Gwo-Shu 
Published: August 2004

ABSTRACT

In trypanosomatids, endocytosis and exocytosis occur exclusively at the flagellar pocket, which represents about 0.43% of the pellicle membrane and is a deep invagination of the plasma membrane where the flagellum extends from the cell. Receptor molecules are selectively retained at the flagellar pocket. We studied the function of clathrin heavy chain (TbCLH) in the trafficking of the flagellar pocket receptors in
Trypanosoma brucei
by using the double-stranded RNA interference approach. It appears that TbCLH is essential for the survival of both the procyclic form and the bloodstream form of
T. brucei
, even though structures resembling large coated endocytic vesicles are absent in procyclic-form trypanosomes. Down-regulation of TbCLH by RNA interference (RNAi) for 24 h rapidly and drastically reduced the uptake of macromolecules via receptor-mediated endocytosis in procyclic-form trypanosomes. This result suggested the importance of TbCLH in receptor-mediated endocytosis of the procyclic-form trypanosome, in which the formation of large coated endocytic vesicles may not be required. Surprisingly, induction of TbCLH RNAi in the procyclic
T. brucei
for a period of 48 h prohibited the export of the flagellar pocket-associated transmembrane receptor CRAM from the endoplasmic reticulum to the flagellar pocket, while trafficking of the glycosylphosphatidylinositol-anchored procyclin coat was not significantly affected. After 72 h of induction of TbCLH RNAi, procyclics exhibited morphological changes to an apolar round shape without a distinct structure of the flagellar pocket and flagellum. Although trypanosomes, like other eukaryotes, use similar organelles and machinery for protein sorting and transport, our studies reveal a novel role for clathrin in the secretory pathway of trypanosomes. We speculate that the clathrin-dependent trafficking of proteins to the flagellar pocket may be essential for the biogenesis and maintenance of the flagellar pocket in trypanosomes.
Mutation of the Regulator of G Protein Signaling Crg1 Increases Virulence inCryptococcus neoformans
Authors: Wang Ping , Cutler Jim , King Jill , Palmer Daniel 
Published: August 2004

ABSTRACTThe regulator of G protein signaling homolog Crg1 was found to be a key regulator of pheromone-responsive mating in the opportunistic human fungal pathogenCryptococcus neoformans. A mutation in theCRG1gene has greatly increased virulence in the prevalently distributedMATα strains of the fungus. Mouse survival time was shortened by 40%, and the lethal dosage was 100-fold less than that of wild-type strains. In addition, the increased virulence ofcrg1mutant strains was dependent on the transcription factor homolog Ste12α but not on the mitogen-activated protein kinase homolog Cpk1. The enhanced mating due toCRG1mutation, however, was still dependent on Cpk1. Interestingly,crg1mutants ofMATα cells produced dark melanin pigment under normally inhibitory conditions, which may relate to the mechanism for increased virulence.
Multiple Triclosan Targets in
Trypanosoma brucei
Authors: Paul Kimberly S. , Bacchi Cyrus J. , Englund Paul T. 
Published: August 2004

ABSTRACT

Trypanosoma brucei
genes encoding putative fatty acid synthesis enzymes are homologous to those encoding type II enzymes found in bacteria and organelles such as chloroplasts and mitochondria. It was therefore not surprising that triclosan, an inhibitor of type II enoyl-acyl carrier protein (enoyl-ACP) reductase, killed both procyclic forms and bloodstream forms of
T. brucei
in culture with 50% effective concentrations (EC
50
s) of 10 and 13 μM, respectively. Triclosan also inhibited cell-free fatty acid synthesis, though much higher concentrations were required (EC
50
s of 100 to 200 μM). Unexpectedly, 100 μM triclosan did not affect the elongation of [
3
H]laurate (C
12:0
) to myristate (C
14:0
) in cultured bloodstream form parasites, suggesting that triclosan killing of trypanosomes may not be through specific inhibition of enoyl-ACP reductase but through some other mechanism. Interestingly, 100 μM triclosan did reduce the level of incorporation of [
3
H]myristate into glycosyl phosphatidylinositol species (GPIs). Furthermore, we found that triclosan inhibited fatty acid remodeling in a cell-free assay in the same concentration range required for killing
T. brucei
in culture. In addition, we found that a similar concentration of triclosan also inhibited the myristate exchange pathway, which resides in a distinct subcellular compartment. However, GPI myristoylation and myristate exchange are specific to the bloodstream form parasite, yet triclosan kills both the bloodstream and procyclic forms. Therefore, triclosan killing may be due to a nonspecific perturbation of subcellular membrane structure leading to dysfunction in sensitive membrane-resident biochemical pathways.
The Forkhead Transcription Factor Fkh2 Regulates the Cell Division Cycle of
Schizosaccharomyces pombe
Authors: Bulmer Richard , Pic-Taylor Aline , Whitehall Simon K. , Martin Kate A. , Millar Jonathan B. A. , Quinn Janet , Morgan Brian A. 
Published: August 2004

ABSTRACT

In eukaryotes the regulation of gene expression plays a key role in controlling cell cycle progression. Here, we demonstrate that a forkhead transcription factor, Fkh2, regulates the periodic expression of
cdc15
+
and
spo12
+
in the M and G
1
phases of the cell division cycle in the fission yeast
Schizosaccharomyces pombe
. We also show that Fkh2 is important for several cell cycle processes, including cell morphology and cell separation, nuclear structure and migration, and mitotic spindle function. We find that the expression of
fkh2
+
is itself regulated in a cell cycle-dependent manner in G
1
coincident with the expression of
cdc18
+
, a Cdc10-regulated gene. However,
fkh2
+
expression is independent of Cdc10 function. Fkh2 was found to be phosphorylated during the cell division cycle, with a timing that suggests that this posttranslational modification is important for
cdc15
+
and
spo12
+
expression. Related forkhead proteins regulate G
2
and M phase-specific gene expression in the evolutionarily distant
Saccharomyces cerevisiae
, suggesting that these proteins play conserved roles in regulating cell cycle processes in eukaryotes.
BANACH SPACE VALUED CAUCHY–RIEMANN EQUATIONS WITH TOTALLY REAL BOUNDARY CONDITIONS
Authors: WEHRHEIM KATRIN 
Published: August 2004

The main purpose of this paper is to give a general regularity result for Cauchy–Riemann equations in complex Banach spaces with totally real boundary conditions. The usual elliptic Lp-regularity results hold true under one crucial assumption: The Banach space is isomorphic to a closed subspace of an Lp-space. (Equivalently, the totally real submanifold is modelled on a closed subspace of an Lp-space.)Secondly, we describe a class of examples of such totally real submanifolds, namely gauge invariant Lagrangian submanifolds in the space of connections over a Riemann surface. These pose natural boundary conditions for the anti-self-duality equation on 4-manifolds with a boundary space-time splitting, leading towards the definition of a Floer homology for 3-manifolds with boundary, which is the first step in a program by Salamon for the proof of the Atiyah–Floer conjecture. The principal part of such a boundary value problem is an example of a Banach space valued Cauchy–Riemann equation with totally real boundary condition.
INVERSE TOPOLOGICAL PRESSURE WITH APPLICATIONS TO HOLOMORPHIC DYNAMICS OF SEVERAL COMPLEX VARIABLES
Authors: MIHAILESCU EUGEN , URBAŃSKI MARIUSZ 
Published: August 2004

In this paper, we introduce a few notions of inverse topological pressure [Formula: see text], defined in terms of backward orbits (prehistories) instead of forward orbits. This inverse topological pressure has some properties similar to the regular (forward) pressure but, in general, if the map is not a homeomorphism, they do not coincide. In fact, there are several ways to define inverse topological pressure; for instance, we show that the Bowen type definition coincides with the one using spanning sets. Then we consider the case of a holomorphic map [Formula: see text] which is Axiom A and such that its critical set does not intersect a particular basic set of saddle type Λ. We will prove that, under a technical condition, the Hausdorff dimension of the intersection between the local stable manifold and the basic set is equal to ts, i.e. [Formula: see text], for all points x belonging to Λ. Here ts represents the unique zero of the function t→P-(tϕs), with P- denoting the inverse topological pressure and [Formula: see text], y∈Λ. In general, [Formula: see text] will be estimated above by ts and below by [Formula: see text], where [Formula: see text] is the unique zero of the map t→P_(tϕs). As a corollary we obtain that, if the stable dimension is non-zero, then Λ must be a non-Jordan curve, and also, if f|Λ happens to be a homeomorphism (like in the examples from [13]), then the stable dimension cannot be zero.
Is the aneuploid chromosome in an apomictic <i>Boechera holboellii</i> a genuine B chromosome?
Authors: Sharbel T.F. , Voigt M.-L. , Mitchell-Olds T. , Kantama L. , de Jong H. 
Published: August 2004

The <i>Boechera holboellii</i> complex comprises <i>B. holboellii</i> and <i>B. drummondii</i>, both of which can reproduce through sex or apomixis. Sexuality is associated with diploidy, whereas apomictic individuals can either be diploid, aneuploid or triploid. Aneuploid individuals are found in geographically and genetically distinct populations and contain a single extra chromosome. It is unknown whether the supernumerary chromosomes are shared by common descent (single origin) or have originated via introgressive hybridizations associated with the repeated transition from diploidy to triploidy. Diploid plants containing the extra chromosome(s) reproduce apomictically, suggesting that the supernumerary elements are associated with apomixis. In this study we compared flow cytometry data, chromosome morphology, and DNA sequences of sexual diploid and apomictic aneuploids in order to establish whether the extra chromosome fits the classical concept of a B chromosome. Karyotype analyses revealed that the supernumerary chromosome in the metaphase complement is heterochromatic and often smaller than the A chromosomes, and differs in length between apomictic plants from different populations. DNA sequence analyses furthermore demonstrated elevated levels of non-synonymous substitutions in one of the alleles, likely that on the aneuploid chromosome. Although the extra chromosome in apomictic <i>Boechera</i> does not go through normal reductional meiosis, in which it may get eliminated or accumulated by a B-chromosome-specific process, its variable size and heterochromatic nature does meet the remaining criteria for a genuine B chromosome in other species. Its prevalence and conserved genetic composition nonetheless implies that this chromosome, if truly a B, may be atypical with respect to its influence on its carriers.
The occurrence of different Bs in <i>Cestrum intermedium</i> and <i>C. strigilatum</i> (Solanaceae) evidenced by chromosome banding
Authors: Fregonezi J.N. , Rocha C. , Torezan J.M.D. , Vanzela A.L.L. 
Published: August 2004

In this study, we examine the morphology, mitotic stability, meiotic behavior and the composition of heterochromatin of B chromosomes in <i>Cestrum intermedium</i> and <i>C. strigilatum</i>. The results showed that B chromosome number shows intraindividual variation in the root meristem, which seems to lead to a slight rate of B elimination in this somatic tissue. B chromosomes in both species were similar in size and shape, but differed with regard to the type, size and distribution of heterochromatin. Possible evolutionary pathways for B chromosome origin in <i>Cestrum</i> are discussed.
Synergistic effect of SU11248 with cytarabine or daunorubicin on FLT3 ITD–positive leukemic cells
Authors: Yee Kevin W. H. , Schittenhelm Marcus , O'Farrell Anne-Marie , Town Ajia R. , McGreevey Laura , Bainbridge Troy , Cherrington Julie M. , Heinrich Michael C. 
Published: August 2004

Abstract
Fetal liver tyrosine kinase 3 internal tandem duplication (FLT3 ITD) mutations are the most common molecular abnormality associated with adult acute myeloid leukemia (AML). To exploit this molecular target, a number of potent and specific FLT3 kinase inhibitors have been developed and are currently being tested in early phase clinical trials of patients with refractory AML. To explore the efficacy of combining a FLT3 inhibitor with standard AML chemotherapy drugs, we tested the effect of combining the FLT3 inhibitor SU11248 with cytarabine or daunorubicin on the proliferation and survival of cell lines expressing either mutant (FLT3 ITD or FLT3 D835V) or wild-type (WT) FLT3. SU11248 had additive-to-synergistic inhibitory effects on FLT3-dependent leukemic cell proliferation when combined with cytarabine or daunorubicin. The synergistic interaction of SU11248 and the traditional antileukemic agents was more pronounced for induction of apoptosis. SU11248 inhibited the proliferation of primary AML myeloblasts expressing mutant FLT3 ITD but not WT FLT3 protein. Combining SU11248 and cytarabine synergistically inhibited the proliferation of primary AML myeloblasts expressing FLT3 ITD but not WT FLT3 protein. These data suggest that the addition of potent FLT3 inhibitors such as SU11248 to AML chemotherapy regimens could result in improved treatment results.
Are interleukin-16 and thrombopoietin new tools for the in vitro generation of dendritic cells?
Authors: Bella Silvia Della , Nicola Stefania , Timofeeva Inna , Villa Maria Luisa , Santoro Armando , Berardi Anna C. 
Published: August 2004

AbstractThe effects of interleukin 16 (IL-16) on dendritic cell (DC) generation from human CD34+ progenitor cells are not known. Here, we show that IL-16 added to a basal cocktail comprised of granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-4, Flt-3 ligand (Flt3L), and tumor necrosis factor α (TNF-α) does induce the CD34+ hematopoietic cells to proliferate in vitro and to differentiate into phenotypically and functionally mature DCs. IL-16 exerts this function more efficiently than stem cell factor (SCF) as a control, thrombopoietin (TPO), or IL-16 plus TPO. Moreover, we show that the combination of IL-16 plus TPO induces the generation of tolerogenic DCs, able to induce an anergic state in T cells that persists when T cells are rechallenged with immunogenic DCs. An altered pattern of cytokine production, a reduced expression of the C-type lectin DC-SIGN, and an increased surface expression of the inhibitory molecules immunoglobulin-like transcript 2 (ILT-2), ILT-3, and ILT-4 may all contribute to confer the tolerogenic properties of these DCs. Generation of tolerogenic DCs may aid the exploration of new therapeutic strategies to promote tolerance to autoantigens and prevent disease development. (Blood. 2004;104:4020-4028)
Ultralarge complexes of PF4 and heparin are central to the pathogenesis of heparin-induced thrombocytopenia
Authors: Rauova Lubica , Poncz Mortimer , McKenzie Steven E. , Reilly Michael P. , Arepally Gowthami , Weisel John W. , Nagaswami Chandrasekaran , Cines Douglas B. , Sachais Bruce S. 
Published: August 2004

AbstractHeparin-induced thrombocytopenia and thrombosis (HITT) is a severe complication of heparin therapy caused by antibodies to complexes between unfractionated heparin (UFH) and platelet factor 4 (PF4) that form over a narrow molar range of reactants and initiate antibody-induced platelet activation. We observed that UFH and tetrameric PF4 formed ultralarge (&gt; 670 kDa) complexes (ULCs) only over a narrow molar range with an optimal ratio of PF4 to heparin of approximately 1:1. These ULCs were stable and visible by electron microscopy, but they could be dissociated into smaller complexes upon addition of heparin. ULCs formed inefficiently when PF4 was incubated with low-molecular-weight heparin, and none formed with the pentasaccharide fondaparinux sodium. In addition, mutation studies showed that formation of ULCs depended on the presence of PF4 tetramers. The ULCs were more reactive as determined by their capacity to bind to a HITT-like monoclonal antibody and showed greater capacity to promote platelet activation in an antibody- and FcγRIIA-dependent manner than were the smaller complexes. The capacity of PF4 to form ULCs composed of multiple PF4 tetramers arrayed in a lattice with several molecules of UFH may play a fundamental role in autoantibody formation, antibody-dependent platelet activation, and the propensity for thrombosis in patients with HITT.
Expression of the AID protein in normal and neoplastic B cells
Authors: Pasqualucci Laura , Guglielmino Roberta , Houldsworth Jane , Mohr Jessica , Aoufouchi Said , Polakiewicz Roberto , Chaganti R. S. K. , Dalla-Favera Riccardo 
Published: August 2004

Abstract
Somatic hypermutation (SHM) targets primarily the immunoglobulin variable region (IgV) genes in germinal center (GC) B cells, thereby allowing antibody affinity maturation. A malfunction of SHM, termed aberrant somatic hypermutation (ASHM), was found in about 50% of diffuse large B-cell lymphomas (DLBCLs), leading to mutations in the 5′ sequences of multiple genes, including oncogenes. Although the SHM mechanism is largely unknown, it was shown to require the activation-induced cytidine deaminase (AID) gene. AID mRNA is expressed in GC B cells and GC-derived lymphomas, but the pattern of expression of the AID protein is not known. Using 2 specific antibodies, here we show that the AID protein can be detected in GC centroblasts and their transformed counterpart (Burkitt lymphoma) but not in pre-GC B cells and post-GC neoplasms, including B-cell chronic lymphocytic leukemia and multiple myeloma. DLBCLs displayed variable levels of AID expression, which did not correlate with IgV ongoing hypermutation, ASHM, or disease subtype. Finally, both in normal and malignant B cells the AID protein appeared predominantly localized in the cytoplasm. These results indicate that the AID protein is specifically expressed in normal and transformed GC B cells; nonetheless, its predominantly cytoplasmic localization suggests that additional mechanisms may regulate its function and may be altered during lymphomagenesis. (Blood. 2004;104:3318-3325)
Similarities in transcription factor IIIC subunits that bind to the posterior regions of internal promoters for RNA polymerase III
Authors: Matsutani Sachiko 
Published: August 2004

Abstract
Background
In eukaryotes, RNA polymerase III (RNAP III) transcribes the genes for small RNAs like tRNAs, 5S rRNA, and several viral RNAs, and short interspersed repetitive elements (SINEs). The genes for these RNAs and SINEs have internal promoters that consist of two regions. These two regions are called the A and B blocks. The multisubunit transcription factor TFIIIC is required for transcription initiation of RNAP III; in transcription of tRNAs, the B-block binding subunit of TFIIIC recognizes a promoter. Although internal promoter sequences are conserved in eukaryotes, no evidence of homology between the B-block binding subunits of vertebrates and yeasts has been reported previously.

Results
Here, I reported the results of PSI-BLAST searches using the B-block binding subunits of human and Shizosacchromyces pombe as queries, showing that the same Arabidopsis proteins were hit with low E-values in both searches. Comparison of the convergent iterative alignments obtained by these PSI-BLAST searches revealed that the vertebrate, yeast, and Arabidopsis proteins have similarities in their N-terminal one-third regions. In these regions, there were three domains with conserved sequence similarities, one located in the N-terminal end region. The N-terminal end region of the B-block binding subunit of Saccharomyces cerevisiae is tentatively identified as a HMG box, which is the DNA binding motif. Although I compared the alignment of the N-terminal end regions of the B-block binding subunits, and their homologs, with that of the HMG boxes, it is not clear whether they are related.

Conclusion
Molecular phylogenetic analyses using the small subunit rRNA and ubiquitous proteins like actin and α-tubulin, show that fungi are more closely related to animals than either is to plants. Interestingly, the results obtained in this study show that, with respect to the B-block binding subunits of TFIIICs, animals appear to be evolutionarily closer to plants than to fungi.