bluuebunny/crossref_metadata_2025_split huggingface dataset:
Postnatal expression and distribution of Refsum disease gene associated protein in the rat retina and visual cortex: effect of binocular visual deprivation
Authors: Ahn Kyu Youn , Nam Kwang Il , Kim Baik Yoon , Cho Chul Woong , Jeong Sang Ki , Yang Kun Jin , Kim Kyung Keun 
Published: October 2002

AbstractPreviously, phytanoyl‐CoA alpha‐hydroxylase‐associated protein 1 (PAHX‐AP1) was isolated as a novel neuron‐specific protein to interact with Refsum disease (RfD) gene PAHX. Its expression in the brain increased after eyelid opening, and the elevated level was maintained through adulthood. In this report, to verify the hypothesis that light could trigger this increase, we have examined the developmental distribution pattern of PAHX‐AP1 in rat retina and visual cortex, and changes of its expression by binocular deprivation. Northern blot analyses demonstrated PAHX‐AP1 expression reached its highest level in the visual cortex and eyeball at 4 weeks after birth, and these levels were maintained through adult life. Two weeks after visual deprivation, its expression in the eyeball and visual cortex decreased compared with the control. In situ hybridization analyses of the retina showed that PAHX‐AP1 expression was limited to the ganglionic cell layer at 10 days after birth, but expressed in the inner nuclear cell layer and extended to the outer nuclear cell layer at 2 and 3 weeks after birth, respectively. Two weeks after visual deprivation, however, it decreased in the ganglionic and inner nuclear cell layer, and disappeared in the rod and cone cell layers. In the visual cortex, strong signals of PAHX‐AP1 were detected in layers IV and VI, and II–VI at 10 days and 2 weeks after birth, respectively. Its expression decreased after 2 weeks of visual deprivation. These results indicate that visual stimulation is essential for the maintenance of PAHX‐AP1 expressions in the retina, especially in the rod and cone cell layers, and visual cortex, and suggest that PAHX‐AP1 may be involved in the developmental regulation of the photoreceptor's function.
What are the reciprocal influences of randomized clinical trials and the patient–psychiatrist relationship?
Authors: Catteau J. , Cyran C. , Bordet R. , Thomas C.E. , Dupuis B.A. 
Published: October 2002

SummaryThe goal of this prospective investigation was to study the course and the quality of patient-psychiatrist relationships during phase II / phase III clinical trials of antidepressant medication prescribed for depressive disorders. All patients who participated in the clinical trials (and subsequently in this survey) signed written informed consent statements and were subject to random double blind treatment assignment. Retrospective analysis of 118 investigations was carried out, and the patients involved were questioned concerning their experiences and impressions during and after the study. Data show that the outcome of clinical trials of antidepressant drugs are not a function of pre-existing good patient-psychiatrist relationships. On the other hand, no effects on the patient-psychiatrist relationship were found as a result of the experimental procedure, and it can be concluded that no detrimental effects on future patient-psychiatrist relationships were incurred.
‘I never even gave it a second thought’: PGCE students' attitudes towards the inclusion of children with speech and language impairments
Authors: Marshall Julie , Stojanovik Vesna , Ralph Sue 
Published: October 2002

AbstractApproximately 7% of young school‐aged children have specific language impairments. Many such children are now being educated in mainstream settings. However, there is a dearth of up‐to‐date and valid research that considers UK (student) teachers' attitudes towards such children. This study aimed to investigate trainee teachers' attitudes towards teaching children with speech and language impairments, to investigate the reported effects of those attitudes on participants' acceptance of teaching such children, and to consider any implications for speech and language therapy (SLT) services and inclusive education. Nineteen trainee teachers (PGCE students) from a university in the North West of England took part in semistructured group interviews. The data were transcribed and analysed qualitatively, and recurrent themes identified. A range of attitudes was expressed, and six major themes were identified from the data. Participants discussed concerns about the resources and knowledge they considered necessary to support fully children with specific language impairments. There was some differentiation of attitudes related to the types of disability that a child may have and the subject being taught. Much of the discussion was about disabilities in general rather than specific to speech and language impairments. Many of the participants reflected on their own previous experiences to inform their opinions. Although many of the participants expressed positive attitudes, some had concerns about workload and at least one was openly hostile to the idea of teaching children with disabilities within mainstream settings. The implications of these findings are discussed in relation to previous research, the quantitative data obtained in this project, SLT services and the increases in the inclusion of children with specific language impairments into mainstream educational settings.
18O Spatial Patterns of Vein Xylem Water, Leaf Water, and Dry Matter in Cotton Leaves
Authors: Gan Kim Suan , Wong Suan Chin , Yong Jean Wan Hong , Farquhar Graham Douglas 
Published: October 2002

Abstract
Three leaf water models (two-pool model, Péclet effect, and string-of-lakes) were assessed for their robustness in predicting leaf water enrichment and its spatial heterogeneity. This was achieved by studying the 18O spatial patterns of vein xylem water, leaf water, and dry matter in cotton (Gossypium hirsutum) leaves grown at different humidities using new experimental approaches. Vein xylem water was collected from intact transpiring cotton leaves by pressurizing the roots in a pressure chamber, whereas the isotopic content of leaf water was determined without extracting it from fresh leaves with the aid of a purpose-designed leaf punch. Our results indicate that veins have a significant degree of lateral exchange with highly enriched leaf water. Vein xylem water is thus slightly, but progressively enriched in the direction of water flow. Leaf water enrichment is dependent on the relative distances from major veins, with water from the marginal and intercostal regions more enriched and that next to veins and near the leaf base more depleted than the Craig-Gordon modeled enrichment of water at the sites of evaporation. The spatial pattern of leaf water enrichment varies with humidity, as expected from the string-of-lakes model. This pattern is also reflected in leaf dry matter. All three models are realistic, but none could fully account for all of the facets of leaf water enrichment. Our findings acknowledge the presence of capacitance in the ground tissues of vein ribs and highlight the essential need to incorporate Péclet effects into the string-of-lakes model when applying it to leaves.
Mutations associated with hemophilia A in the 558-565 loop of the factor VIIIa A2 subunit alter the catalytic activity of the factor Xase complex
Authors: Jenkins P. Vincent , Freas Jan , Schmidt Kyla M. , Zhou Qian , Fay Philip J. 
Published: October 2002

AbstractThe 558-565 loop region in the A2 subunit of factor (F) VIIIa forms a direct interface with FIXa. We have expressed and purified B-domainless FVIII (FVIIIWT) and B-domainless FVIII containing the hemophilia A–associated mutations Ser558Phe, Val559Ala, Asp560Ala, Gln565Arg, and the activated protein C cleavage site mutant Arg562Ala. Titration of FVIIIa in FXa generation assays showed that the mutant and wild-type proteins had similar functional affinities for FIXa (dissociation constant [Kd] values ∼5 nM-20 nM and ∼100 nM-250 nM in the presence and absence of phospholipid, respectively). The catalytic activities of the factor Xase complex composed of the hemophilia A–associated FVIII species were markedly reduced both in the presence and absence of phospholipid. FVIIIWT and FVIIIArg562Ala showed catalytic rate constant (kcat) values of approximately 60 minute−1 in the presence of phospholipid, whereas the hemophilia A–associated mutants showedkcat values ranging from 3.3 minute−1 to 7.5 minute−1. In the absence of phospholipid, all kcat values were reduced but FVIIIWT and FVIIIArg562Ala retained higher activities as compared with the hemophilic mutant FVIII forms. Fluorescence anisotropy experiments using fluorescein-modified FIXa confirmed that all FVIII forms interacted with FIXa. However, the presence of factor X yielded minimal increases in anisotropy observed with the mutant factor VIII forms, consistent with their reduced activity. These results show that residues within the 558-565 loop are critical in modulating FIXa enzymatic activity but do not contribute significantly to the affinity of FVIIIa for FIXa.
Complete allogeneic hematopoietic chimerism achieved by a combined strategy of in utero hematopoietic stem cell transplantation and postnatal donor lymphocyte infusion
Authors: Hayashi Satoshi , Peranteau William H. , Shaaban Aimen F. , Flake Alan W. 
Published: October 2002

AbstractIn utero hematopoietic stem cell transplantation (IUHSCTx) can achieve mixed hematopoietic chimerism and donor-specific tolerance without cytoreductive conditioning or immunosuppression. The primary limitation to the clinical application of IUHSCTx has been minimal donor cell engraftment, well below therapeutic levels for most target diseases. Donor lymphocyte infusion (DLI) has been used in postnatal circumstances of mixed chimerism as targeted immunotherapy to achieve a graft-versus-hematopoietic effect and to increase levels of donor cell engraftment. In this report we demonstrate in the murine model that a combined approach of IUHSCTx followed by postnatal DLI can convert low-level, mixed hematopoietic chimerism to complete donor chimerism across full major histocompatibility complex barriers with minimal risk for graft-versus-host disease (GVHD). Time-dated embryonic day 14 (E14) to E15 Balb/c (H-2Kd, CD45.2) fetuses underwent intraperitoneal injection of 5 × 106T-cell–depleted B6 (H-2Kb, CD45.2) bone marrow cells. Chimeric recipients then received transplants at either 4 or 8 weeks of age with 1 of 3 doses (5, 15, or 30 × 106cells) of donor congenic splenocytes (B6-Ly5.2/Cr, H-2Kb, CD45.1). The response to DLI was dose dependent, with conversion to complete donor peripheral blood chimerism in 100% of animals that received high-dose (30 × 106 cells) DLI. Only 1 of 56 animals receiving this dose succumbed to GVHD. This study directly supports the potential therapeutic strategy of prenatal tolerance induction to facilitate nontoxic postnatal cellular therapy and organ transplantation, and it has broad implications for the potential treatment of prenatally diagnosed genetic disorders.
Influence of plasma immunoglobulin level on antibody synthesis
Authors: Goldammer Andreas , Derfler Kurt , Herkner Kurt , Bradwell Arthur R. , Hörl Walter H. , Haas Martin 
Published: October 2002

Abstract
In previous experimental animal studies it has been demonstrated that antibody depletion is not followed by increased antibody synthesis. To assess whether these results are conferrable to antibody-depleted humans, we measured free light chains (flcs) as markers of current antibody synthesis in 8 patients treated with immunoadsorption (IA) therapy. Specific and bulk immunoglobulin levels were obtained simultaneously. The mean serum flc concentration increased to the preapheresis value within 1 day and remained unchanged thereafter. Total immunoglobulin G (IgG) and specific antibody concentrations increased to pretreatment values in 88% and 43% of the patients, respectively, and remained below the original values in the others. In conclusion, the lack of increased flc synthesis after IA confirms the absence of a feedback mechanism regulating antibody synthesis. The restoration of serum IgG levels after IA, therefore, does not result from increased antibody synthesis but is probably related to changes of catabolism and immunoglobulin backflow.
Vesicle-associated membrane protein 3 (VAMP-3) and VAMP-8 are present in human platelets and are required for granule secretion
Authors: Polgár János , Chung Sul-Hee , Reed Guy L. 
Published: October 2002

AbstractSecretion of platelet granules is necessary for normal hemostasis. Platelet secretion requires soluble N-ethylmaleimide–sensitive factor attachment protein (SNAP) receptor (SNARE) complex formation between different members of the syntaxin, SNAP-25, and vesicle-associated membrane protein (VAMP) gene families. Using microcapillary reverse-phase high-performance liquid chromatography–nano-electrospray tandem mass spectrometry, we identified VAMP-3 and VAMP-8 as VAMP isoforms coimmunoprecipitated from platelets with syntaxin 4. Immunoblotting experiments confirmed the presence of VAMP-3 and VAMP-8 but not VAMP-1 or VAMP-2 in platelets. To examine the effect of VAMP proteins on platelet secretion, soluble recombinant (r) VAMP-2, rVAMP-3, and rVAMP-8 were incubated with streptolysin O–permeabilized platelets. Secretion of α granules (monitored by flow cytometric measurement of P-selectin) was blocked, and dense-granule secretion (assessed by release of carbon 14–serotonin) was almost completely inhibited by rVAMP-3, whereas rVAMP-8 inhibited secretion of dense granules but not α granules. In contrast, rVAMP-2, which formed SNARE complexes in vitro, had no effect on platelet exocytosis. We conclude that VAMP-3 and VAMP-8 form SNARE complexes with platelet syntaxin 4 and are required for platelet granule secretion.
Lentiviral vectors containing the human immunodeficiency virus type-1 central polypurine tract can efficiently transduce nondividing hepatocytes and antigen-presenting cells in vivo
Authors: VandenDriessche Thierry , Thorrez Lieven , Naldini Luigi , Follenzi Antonia , Moons Lieve , Berneman Zwi , Collen Desire , Chuah Marinee K. L. 
Published: October 2002

Abstract
High-titer self-inactivating human immunodeficiency virus type-1 (HIV-1)–based vectors expressing the green fluorescent protein reporter gene that contained the central polypurine and termination tract and the woodchuck hepatitis virus posttranscriptional regulatory element were constructed. Transduction efficiency and biodistribution were determined, following systemic administration of these improved lentiviral vectors. In adult severe combined immunodeficiency (SCID) mice, efficient stable gene transfer was achieved in the liver (8.0% ± 6.0%) and spleen (24% ± 3%). Most transduced hepatocytes and nonhepatocytes were nondividing, thereby obviating the need to induce liver cell proliferation. In vivo gene transfer with this improved lentiviral vector was relatively safe since liver enzyme concentration in the plasma was only moderately and transiently elevated. In addition, nondividing major histocompatibility complex class II–positive splenic antigen-presenting cells (APCs) were efficiently transduced in SCID and normal mice. Furthermore, B cells were efficiently transduced, whereas T cells were refractory to lentiviral transduction in vivo. However, in neonatal recipients, lentiviral transduction was more widespread and included not only hepatocytes and splenic APCs but also cardiomyocytes. The present study suggests potential uses of improved lentiviral vectors for gene therapy of genetic blood disorders resulting from serum protein deficiencies, such as hemophilia, and hepatic disease. However, the use of liver-specific promoters may be warranted to circumvent inadvertent transgene expression in APCs. In addition, these improved lentiviral vectors could potentially be useful for genetic vaccination and treatment of perinatal cardiac disorders.
The Stat5-RARα fusion protein represses transcription and differentiation through interaction with a corepressor complex
Authors: Maurer Alexander B. , Wichmann Christian , Gross Alexandra , Kunkel Hana , Heinzel Thorsten , Ruthardt Martin , Groner Bernd , Grez Manuel 
Published: October 2002

Abstract
The transcription factor Stat5 mediates the cellular response to activation of multiple cytokine receptors involved in the regulation of proliferation and differentiation of hematopoietic cells. Recently, the human Stat5 gene was found to be translocated to the RARα gene in a patient with acute promyelocytic leukemia indicating that Stat5 might also play a role in cellular transformation. We investigated the mechanism by which Stat5 might exert this function and studied the biochemical and cellular functions of fusion proteins comprising Stat5 and RARα. The expression of Stat5-RARα causes the transcriptional repression of gene transcription, a process that requires the coiled-coil domain of Stat5 (amino acid positions 133-333). Oligomerization of this domain in the Stat5-RARα fusion protein leads to stable binding of the corepressor SMRT independent of all-trans retinoic acid (ATRA) stimulation and is accompanied by an impaired response to differentiation signals in hematopoietic cells. This inhibitory effect on myeloid differentiation cannot be overcome by simultaneous coexpression of RARα. We conclude that Stat5 is capable of interacting with a corepressor complex that alters the pattern of corepressor binding to RARα and its dissociation in response to ATRA stimulation, leading to enhanced repressor activity and a block of hematopoietic differentiation.
Intrathymic and extrathymic development of human plasmacytoid dendritic cell precursors in vivo
Authors: Weijer Kees , Uittenbogaart Christel H. , Voordouw Arie , Couwenberg Franka , Seppen Jurgen , Blom Bianca , Vyth-Dreese Florry A. , Spits Hergen 
Published: October 2002

AbstractThe development of plasmacytoid dendritic cells (pDC2) from human CD34+ stem cells in vivo was studied in RAG-2−/− interleukin (IL)-2Rγ−/− mice that lack functional T and B cells and natural killer cells. CD34+ cells isolated from fetal liver or thymus were labeled with 5- and 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) and were injected into a human thymus grafted subcutaneously in the RAG-2−/− IL-2Rγ−/− mice. One to 4 weeks later the CFSE label was found not only in T cells but also in CD123+/high CD4+CD45RA+ pDC2, indicating that the CD34+ cells can develop into pDC2 within a thymus. In addition to pDC2, CFSE-labeled dendritic cells with a mature phenotype, determined by the cell surface markers CD11c, CD83, and CD80, were found in the injected human thymus graft. pDC2 was not found in the periphery of mice carrying a human thymic graft, indicating that the intrathymic pDC2 failed to emigrate from the thymus. We also demonstrate that pDC2 can develop outside the thymus because relatively high percentages of pDC2 were found in the periphery after the intravenous injection of CD34+CD38−fetal liver cells in RAG-2−/− IL-2Rγ−/−mice without a human thymus graft. These data indicate that the thymus and the peripheral pDC2 develop independently of each other.
Identification of a myeloid intrathymic pathway of dendritic cell development marked by expression of the granulocyte macrophage–colony-stimulating factor receptor
Authors: de Yébenes Virginia G. , Carrasco Yolanda R. , Ramiro Almudena R. , Toribio Marı́a L. 
Published: October 2002

AbstractIn this study, the finding that a significant proportion of all dendritic cells (DCs) resident in vivo in the human postnatal thymus displayed a myeloid-related phenotype prompted us to re-examine the developmental origin of thymic DCs, a cell type hitherto considered to represent a homogeneous lymphoid-derived population. We show here that these novel intrathymic DCs are truly myeloid, as they arise from CD34+ early thymic progenitors through CD34lointermediates which have lost the capacity to generate T cells, but display myelomonocytic differentiation potential. We also demonstrate that phenotypically and functionally equivalent myeloid precursors devoid of T-cell potential do exist in vivo in the postnatal thymus. Moreover, although interleukin 7 (IL-7) supports the generation of such myeloid intermediates, we show that their developmental branching from the main intrathymic T-cell pathway is linked to the up-regulation of the myelomonocytic granulocyte macrophage–colony-stimulating factor (GM-CSF) receptor, to the down-regulation of the IL-7 receptor and to the lack of pre–T-cell receptor α (pTα) gene transcriptional activation. Taken together, these data challenge the current view that the thymus is colonized by a lymphoid-restricted progenitor and provide evidence that a more immature precursor population with lymphoid and myelomonocytic potential is actually seeding the human postnatal thymus.
Caspase-catalyzed cleavage and activation of Mst1 correlates with eosinophil but not neutrophil apoptosis
Authors: De Souza Patricia M. , Kankaanranta Hannu , Michael Ashour , Barnes Peter J. , Giembycz Mark A. , Lindsay Mark A. 
Published: October 2002

AbstractWe have examined the role of caspase-mediated cleavage of the Ste20-like kinases, mammalian sterile 20-like 1 and 2 (Mst1/Mst2), in the mechanism of human eosinophil and neutrophil apoptosis. Initial measurements of kinase activity, using myelin basic protein (MBP) as a substrate in “in-gel” renaturation assays, showed that constitutive eosinophil and neutrophil apoptosis were associated temporally with the activation of a 36-kd MBP kinase (p36 MBPK) and a 34-kd MBP kinase (p34 MBPK), respectively. A constitutively active 63-kd MBP kinase (p63 MBPK) was also detected in freshly prepared eosinophils but not neutrophils, whose activity was transiently augmented during spontaneous apoptosis. Immunoblotting studies demonstrated the expression of Mst1 and Mst2 in eosinophils but not neutrophils whereas immunoprecipitation studies identified the p63 MBPK activity as being Mst1 and Mst2 and showed that the p36 MBPK activity represented the N-terminal catalytic fragment of Mst1. A role for the p36 MBPK in eosinophil cell death was supported by studies showing increased activation upon exposure to the proapoptotic Fas/CD95-activating antibody, CH-11, and attenuation in the presence of the survival-promoting cytokine, interleukin-5. Furthermore, spontaneous and Fas-induced activation of p36 MBPK was inhibited by catalase and the general caspase inhibitor, z-Asp-CH2-DCB, at concentrations that suppressed eosinophil apoptosis. These studies therefore implicate a role for caspase- and H2O2-mediated cleavage of the Mst1 and the subsequent release of the 36-kd catalytic fragment in the mechanism of eosinophil apoptosis. In contrast, neutrophil apoptosis occurs independently of Mst1 and Mst2 but instead is correlated with the activation of an as-yet-unidentified 34-kd MBPK.