bluuebunny/crossref_metadata_2025_split huggingface dataset:
Targeting lentiviral vector expression to hepatocytes limits transgene-specific immune response and establishes long-term expression of human antihemophilic factor IX in mice
Authors: Follenzi Antonia , Battaglia Manuela , Lombardo Angelo , Annoni Andrea , Roncarolo Maria Grazia , Naldini Luigi 
Published: January 2004

Abstract
Stable gene replacement by in vivo administration of lentiviral vectors (LVs) has therapeutic potential for metabolic disorders and other systemic diseases. We studied the expression of intracellular and secreted proteins by LVs in immunocompetent mice. Liver, spleen, and bone marrow cells were efficiently transduced. However, transgene expression, driven by a ubiquitous promoter, was limited by transgene-specific cellular and humoral immune responses, leading to the clearance of transduced cells. After green fluorescent protein (GFP) gene transfer, the liver showed infiltration of CD8+ cytotoxic T cells, and GFP-specific CD8+ T cells were isolated from the spleen. After human factor IX (hF.IX) gene transfer, anti-hF.IX antibodies were induced. These immune responses were not detected in mice injected with heat-inactivated or genome-lacking LVs or in GFP-transgenic mice, indicating that they were specifically triggered by transgene expression in vivo. Intriguingly, selective targeting of LV expression to hepatocytes limited the immune responses to the transgenes. By this approach, high levels of hF.IX, potentially in the therapeutic range, were reached and maintained long term in immunocompetent mice, without inducing antibody formation. These results prompt further studies in relevant animal models to explore the potential of in vivo LV administration for the gene therapy of hemophilias and other liver-based diseases.
Gene expression in human embryonic stem cell lines: unique molecular signature
Authors: Bhattacharya Bhaskar , Miura Takumi , Brandenberger Ralph , Mejido Josef , Luo Yongquan , Yang Amy X. , Joshi Bharat H. , Ginis Irene , Thies R. Scott , Amit Michal , Lyons Ian , Condie Brian G. , Itskovitz-Eldor Joseph , Rao Mahendra S. , Puri Raj K. 
Published: January 2004

Abstract
Human embryonic stem (huES) cells have the ability to differentiate into a variety of cell lineages and potentially provide a source of differentiated cells for many therapeutic uses. However, little is known about the mechanism of differentiation of huES cells and factors regulating cell development. We have used high-quality microarrays containing 16 659 seventy–base pair oligonucleotides to examine gene expression in 6 of the 11 available huES cell lines. Expression was compared against pooled RNA from multiple tissues (universal RNA) and genes enriched in huES cells were identified. All 6 cell lines expressed multiple markers of the undifferentiated state and shared significant homology in gene expression (overall similarity coefficient &gt; 0.85).A common subset of 92 genes was identified that included Nanog, GTCM-1, connexin 43 (GJA1), oct-4, and TDGF1 (cripto). Gene expression was confirmed by a variety of techniques including comparison with databases, reverse transcriptase–polymerase chain reaction, focused cDNA microarrays, and immunocytochemistry. Comparison with published “stemness” genes revealed a limited overlap, suggesting little similarity with other stem cell populations. Several novel ES cell–specific expressed sequence tags were identified and mapped to the human genome. These results represent the first detailed characterization of undifferentiated huES cells and provide a unique set of markers to profile and better understand the biology of huES cells. (Blood. 2004;103: 2956-2964)
Interleukin-6 deficiency affects bone marrow stromal precursors, resulting in defective hematopoietic support
Authors: del Carmen Rodríguez María , Bernad Antonio , Aracil Miguel 
Published: January 2004

AbstractInterleukin-6 (IL-6) is a critical factor in the regulation of stromal function and hematopoiesis. In vivo bromodeoxyuridine incorporation analysis indicates that the percentage of Lin-Sca-1+ hematopoietic progenitors undergoing DNA synthesis is diminished in IL-6-deficient (IL-6-/-) bone marrow (BM) compared with wild-type BM. Reduced proliferation of IL-6-/- BM progenitors is also observed in IL-6-/- long-term BM cultures, which show defective hematopoietic support as measured by production of total cells, granulocyte macrophage-colony-forming units (CFU-GMs), and erythroid burst-forming units (BFU-Es). Seeding experiments of wild-type and IL-6-/- BM cells on irradiated wild-type or IL-6-deficient stroma indicate that the hematopoietic defect can be attributed to the stromal and not to the hematopoietic component. In IL-6-/- BM, stromal mesenchymal precursors, fibroblast CFUs (CFU-Fs), and stroma-initiating cells (SICs) are reduced to almost 50% of the wild-type BM value. Moreover, IL-6-/- stromata show increased CD34 and CD49e expression and reduced expression of the membrane antigens vascular cell adhesion molecule-1 (VCAM-1), Sca-1, CD49f, and Thy1. These data strongly suggest that IL-6 is an in vivo growth factor for mesenchymal precursors, which are in part implicated in the reduced longevity of the long-term repopulating stem cell compartment of IL-6-/- mice. (Blood. 2004;103:3349-3354)
Safety and efficacy of the monoclonal anti–interleukin-5 antibody SCH55700 in the treatment of patients with hypereosinophilic syndrome
Authors: Klion Amy D. , Law Melissa A. , Noel Pierre , Kim Yae-Jean , Haverty Thomas P. , Nutman Thomas B. 
Published: December 2003

Abstract
Four patients with hypereosinophilic syndrome (HES) refractory to or intolerant of treatment with conventional therapy were treated with a single 1 mg/kg dose of SCH55700. SCH55700 was extremely well tolerated. Two of the 4 patients responded with a fall in eosinophil counts to within the normal range within 48 hours of receiving the drug, accompanied by marked improvement in clinical signs and symptoms. Response was not predicted by serum interleukin-5 (IL-5) levels or presence of the FIP1L1/PDGFRA mutation. Eosinophil counts remained suppressed for up to 12 weeks after treatment; however, exacerbation of symptoms and eosinophilia above baseline levels occurred as drug levels waned. Reinstitution of treatment with monthly SCH55700 led to decreased eosinophilia and symptomatic improvement, albeit to a lesser degree than that seen after the initial dose. These data suggest that anti–IL-5 therapy may be useful in the treatment of HES irrespective of the underlying etiology, although the observed rebound eosinophilia and attenuation of response require further study. (Blood. 2004;103:2939-2941)
Down-regulation of Hox A7 is required for cell adhesion and migration on fibronectin during early HL-60 monocytic differentiation
Authors: Leroy Pascale , Berto Fréderick , Bourget Isabelle , Rossi Bernard 
Published: January 2004

AbstractHox genes, which are key regulators of cell fate and pattern formation during embryogenesis, are also important regulators of hematopoiesis, and different combinations of Hox gene products are involved in lineage commitment or maturation. However, their molecular and cellular modes of action are not yet completely understood. Recent studies have indicated that Hox genes are involved in the regulation of cell–extracellular matrix (ECM) interactions and cell migration. Here, we report that Hox A7, a gene frequently overexpressed in acute myeloid leukemia, is down-regulated during HL-60 monocytic differentiation. Using a model in which HL-60 cells are induced to differentiate toward the monocytic lineage with bone marrow stromal-like cells, we demonstrate that Hox A7-sustained expression disturbs the regulation of cell adhesive and migratory capacities on fibronectin during early differentiation. We show that this is accompanied by a partial blockage of the transcriptional induction of proline-rich tyrosine kinase 2, a gene coding for a focal adhesion kinase active in monocytes, and of tissue transglutaminase, a gene coding for a fibronectin coreceptor in monocytes. This is the first report that demonstrates the involvement of a Hox gene in the regulation of adhesion and migration of hematopoietic cells and that links it to the deregulation of genes involved in cell–ECM interactions and downstream signaling pathways.
Expression and synthesis of fibroblast growth factor-9 in human γδ T-lymphocytes. Response to isopentenyl pyrophosphate and TGF-β1/IL-15
Authors: Workalemahu Grefachew , Foerster Martin , Kroegel Claus 
Published: January 2004

Abstractγδ T-lymphocytes are believed to play a role in maintaining the normal configuration of epithelial tissue. As little is known about the factors mediating this function, we addressed the question of whether γδ T-lymphocytes produce fibroblast growth factor (FGF)-9 as well as two other growth factors associated with epithelial tissue reconstitution. Blood γδ T cells isolated from healthy donors were grown in the presence of isopentenyl pyrophosphate (IPP) or transforming growth factor-β1 (TGF-β1)/interleukin-15 (IL-15) for 24 h and were assessed for the expression and synthesis of FGF-9, keratinocyte growth factor (KGF), and epidermal growth factor (EGF). Resting human γδ T cells constitutively expressed KGF and FGF-9 mRNA but no EGF mRNA. In the presence of IPP, FGF-9 mRNA expression significantly increased in a dose-dependent manner, expression of KGF remained unaltered, and EGF mRNA could not be detected. In contrast to IPP, stimulation of the cells with TGF-β1/IL-15 did not alter FGF-9 expression. Moreover, stimulation with anti-CD3 does not induce FGF-9 expression but triggers a high signal of interferon-γ mRNA. Western blot analysis of γδ T cell lysates, prepared 4 days following stimulation with IPP, showed an increase of FGF-9 protein as compared with control cells. In conclusion, the results demonstrate for the first time that human blood and bronchoalveolar lavage γδ T-lymphocytes are capable of expressing FGF-9. The data also provide novel evidence that immunoregulatory cells can synthesize FGF-9.
Limitations with in vitro production of dendritic cells using cytokines
Authors: O’Neill Helen C , Wilson Heather L 
Published: January 2004

AbstractDendritic cells (DC) are the most effective antigen-presenting cells. Many studies now show that DC can be generated in vitro from a number of starting cell populations containing hematopoietic precursors. The protocols used involve different combinations of cytokines including granulocyte macrophage-colony stimulating factor (GM-CSF), which supports myeloid precursors, or interleukin-7, which supports lymphoid precursors. DC are commonly generated by in vitro culture of bone marrow or monocytes with GM-CSF and other cytokines. However, these cultures do not sustain DC production for long periods of time and do not allow the identification or study of intermediate stages in cell development. In vitro cytokine-dependent cultures of DC precursors do provide a reliable source of DC for stimulating immune responses. However, use of cells produced in cytokine-dependent cultures for the study of DC differentiation is limited, as DC development in vivo differs in cytokine dependency.
Effects of cholera toxin on innate and adaptive immunity and its application as an immunomodulatory agent
Authors: Lavelle Ed C , Jarnicki Andrew , McNeela Edel , Armstrong Michelle E , Higgins Sarah C , Leavy Olive , Mills Kingston H G 
Published: January 2004

AbstractCholera toxin (CT) is a potent vaccine adjuvant when administered via parenteral, mucosal, or transcutaneous routes. It also inhibits innate inflammatory responses induced by pathogen-derived molecules, such as lipopolysaccharide (LPS). We demonstrated previously that CT promotes the induction of regulatory type 1 T cells (Tr1) as well as T helper type 2 cells (Th2). T cells from mice immunized with antigen in the presence of CT produced high levels of interleukin (IL)-10 and IL-5 and low levels of IL-4 and interferon-γ (IFN-γ). Here, we demonstrate that immunization with antigen in the presence of CT induced a population of antigen-specific CD4+ T cells that produced IL-10 in the absence of IL-4, in addition to cells that coexpressed IL-4 and IL-10 or produced IL-4 only. CT-generated Tr1 cells inhibited antigen-specific proliferation as well as IFN-γ production by Th1 cells, and this suppression was cell contact-independent. It is interesting that coincubation with Th1 cells significantly enhanced IL-10 production by the Tr1 cells. As IL-10 can promote the differentiation of Tr1 cells, we investigated cytokine production by dendritic cells (DC) following exposure to CT. Previous data showed that CT can modulate the expression of costimulatory molecules and inhibit the production of chemokines and cytokines, including IL-12 and tumor necrosis factor α and enhance IL-10 production. Here, we show that CT synergizes with LPS to induce IL-6 and IL-1β in addition to IL-10 production by immature DC. Therefore, CT may promote the induction of Th2 and Tr1 cells in part via selective modulation of DC cytokine production and costimulatory molecule expression.
Is hand splinting effective for adults following stroke? A systematic review and methodological critique of published research
Authors: Lannin N A , Herbert R D 
Published: January 2004

Background: Upper limb hemiplegia after stroke is common and disabling. Hand splints are widely used to prevent contracture and reduce spasticity. Objective: To assess the effectiveness of hand splinting on the hemiplegic upper extremity following stroke. Search strategy: A search was conducted of the Cochrane Central Register of Controlled Trials; the electronic databases MEDLINE, EMBASE, CINAHL, PEDro, SCI, SSCI; websites of professional associations; reference lists in trial reports and other relevant articles. Selection criteria: Studies of the effect of upper extremity splinting on motor control, functional abilities, contracture, spasticity, or pain in the hand or wrist. Data collection and analysis: Validity of studies was assessed systematically and a content analysis was conducted of the methodologies used. Methodological quality of randomized trials was rated by two independent assessors using the PEDro scale. Results: Nineteen studies were appraised for content. Of these, most (63%) were reports of case series. Four studies (21%) were randomized controlled trials. Methodological scores of trials ranged from 2 to 8 (maximum possible score 10). One trial of nominally ‘medium’ quality reported that in‘atable arm splinting makes no difference to hand function (mean difference on Fugl-Meyer Assessment –0.12, 95% confidence interval (CI) –9.8 to 9.6). The remaining trials investigated effects of thermoplastic splints; one trial of ‘high quality’ reported no difference in contracture formation in the wrist and finger ‘exor muscles after wearing a hand splint which positioned the wrist in the traditional functional position for 12 hours each night for four weeks (mean difference in range of movement after four weeks was 1°, 95% CI –3.7° to 6.1°; power >80%). All remaining trials were of poor methodological quality. Limited research and lack of a no-splint control group in all trials to date limit the usefulness of these results. Reviewer's conclusion: There is insufficient evidence to either support or refute the effectiveness of hand splinting for adults following stroke.
The effect of a slider shoe on hemiplegic gait
Authors: Cross J , Tyson S F 
Published: January 2004

Objective: To assess the effect of a slider shoe on the gait speed and energy efficiency of hemiplegic gait. Design: A–B–A single-case design to compare walking with and without the slider shoe. Results were assessed graphically using the 2SD method. Setting: Stroke unit of an NHS general hospital in the UK. Subjects: Four acute stroke patients undergoing gait rehabilitation. Intervention: Walking practice with and without a slider shoe worn over the real shoe of the weak leg. Main outcome measures: Gait speed (10-m walk test) and energy efficiency (Physiological Cost Index). Results: All subjects showed an improvement in speed and efficiency when wearing the slider shoe compared with the baseline phase (A1). Three subjects showed a sustained improvement in efficiency and two showed a sustained improvement in speed in the second baseline phase (A2). Conclusion: A slider shoe may improve the speed and efficiency of hemiplegic gait for people in the early stages of gait rehabilitation. Further investigation is warranted.
Reliability and validity of a postal version of the Reintegration to Normal Living Index, modified for use with stroke patients
Authors: Daneski K , Coshall C , Tillingand K , Wolfe C D A 
Published: January 2004

Objective: To adapt the Reintegration to Normal Living Index (RNLI) for postal use with stroke patients. Design: Reliability was examined using test–retest on 26 stroke patients. Construct validity was tested on 76 patients by examining correlations between the modified RNLI and related scales. Subjects: Patients at three months to one year post stroke. Results: All items demonstrated better than chance agreement between test and retest and seven items substantial agreement (kappa= >0.61). The modified RNLI correlated positively with related scales. Patients with stronger reintegration to normal living had better outcomes in anxiety, depression, daily activity and quality of life. Conclusion: This postal instrument appears reliable and valid and may be a useful outcome measure in stroke studies.
Assessing mood in patients with multiple sclerosis
Authors: Groom Madeleine J , Lincoln Nadina B , Francis Valerie M , Stephan Talal F 
Published: January 2004

Objective: To assess the validity and reliability of mood measures for use with people with multiple sclerosis (MS). Design: Four mood measures designed for use with people with communication and cognitive problems were completed; two were completed with the patient at interview and two by hospital staff or carers of the patients. This procedure was repeated two weeks later to assess test–retest reliability. Subjects: People with MS at a rehabilitation unit ( n = 22) and living in the community ( n = 27). Main outcome measures: The Stroke Aphasic Depression Questionnaire (SADQ) and Signs of Depression Scale (SODS) were completed by rehabilitation unit staff or by carers of the patients. An assistant psychologist completed the Visual Analogue Self-Esteem Scale (VASES) and the Visual Analogue Mood Scales (VAMS) with each patient. The Guy's Neurological Disability Scale (GNDS) was administered to assess physical disability. Nonprofessional carers were asked to complete the General Health Questionnaire 12 (GHQ-12) about themselves. Results: In both samples there were significant correlations between scores on the two self-report measures (VASES and VAMS) ( rs = 0.51–0.79) and between scores on the two observational measures (SADQ and SODS) ( rs = 0.62–0.81) but not between the observational and self-report measures. There was a significant correlation between the SODS and the carer GHQ ( rs = 0.68, p < 0.01), indicating that carer mood infuenced reporting of patients' mood. There was no significant infuenced of profession on the observational measures, indicating good inter-rater reliability. Test–retest reliability was high for patients in the community but not for patients in hospital. Conclusions: In both the inpatient and community samples, the self-report and observational mood measures appeared to be measuring different aspects of patients' mood. Further investigation is needed to determine which of these is providing the most accurate information.
Effects of knee joint angle and tilt table incline on force distribution at the feet and supporting straps
Authors: Morgan Claire L , Cullen Gerard P , Stokes Maria , Swan Anthony V 
Published: January 2004

Objective: To investigate the effects of tilt table incline and knee flexion angle on the degree of weight bearing and forces exerted across the supporting straps. Design: A quantitative and exploratory study to investigate the effects of a mechanical procedure. Setting: Physiotherapy gymnasium. Subjects: Twelve healthy subjects (9 female, 3 male) aged 22–45 years. Interventions: Subjects stood on a tilt table, on two occasions, with simulated contractures of 10° and 40° knee flexion. Nine tilt angles, between 10° and 90°, were maintained for 1 minute each in random order. Main outcome measures: Force was recorded from single-point load cells placed under the feet, and at knee and chest straps. Results: The degree of simulated knee contracture (10° or 40°) influenced the distribution of forces at different recording sites. Weight bearing increased with table incline and was significantly less with the 40° than the 10° knee angle ( p < 0.001). Conversely, forces across the knee straps were systemically higher with the 40° knee angle ( p < 0.0001). The effects were accentuated by greater body weight. Forces across the chest strap also increased with tilt and were significantly larger with the 40° knee angle ( p < 0.05). Conclusions: The relationships between table incline, angle of knee flexion and distribution of forces generated during tilt table standing have been quantified and described. Standing with flexed knees involved less weight bearing under the feet and greater force exerted across the supporting straps. These effects were more pronounced at the higher knee angle and with greater body weight, and could be modified by reducing table incline.
Prevalence of pressure sores in a community sample of spinal injury patients
Authors: Raghavan P , Raza W A , Ahmed Y S , Chamberlain M A 
Published: January 2004

Objectives: To estimate the point prevalence of pressure sores in a community sample of spinal cord injured patients who were followed up by a spinal injuries unit and to evaluate whether self-management strategies were associated with decreased risk of pressure sores. Setting: A regional spinal injuries unit, UK. Design: Postal questionnaire survey. Main outcome measure: Presence of pressure sores. Subjects: All patients who were being followed up on a regular basis by the unit. Results: Out of 760, 520 replied to the questionnaire; 472 were eligible for analysis. Point prevalence of pressure sores was 23% (99). Failure to inspect the skin daily for pressure damage was associated with decreased prevalence of pressure sores (odds ratio (OR) 0.5; 95% confidence interval (CI) 0.2–0.83). Those who inspected their skin daily, however, had a higher proportion of stage I pressure sores, but this was not statistically significant. Smoking (OR 1.8; 95% CI 1–3.3) and pre-existing medical problems (OR 1.8; 95% CI 1–3) were associated with increased prevalence of pressure sores. Regular lifting of weight at least once in an hour while seated, age, gender, neurological level, employment status, living alone and faecal and urinary incontinence were not significant predictors of pressure sores. Conclusions: Nearly one-quarter of participants had pressure sores at the time of the survey. Periodic weight lifts and daily inspection of skin for pressure damage were not associated with decreased prevalence of pressure sores in this sample. However, those who inspected skin daily tended to detect pressure damage early.