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排序方式: 共有43条查询结果,搜索用时 31 毫秒
1.
Steenbergen EJ; Verhagen OJ; van Leeuwen EF; van den Berg H; von dem Borne AE; van der Schoot CE 《Blood》1995,86(2):692-702
Crosslineage T-cell receptor delta (TCR delta) rearrangements are widely used as tumor markers for the follow up of minimal residual disease in childhood B-precursor acute lymphoblastic leukemia (ALL) by polymerase chain reaction (PCR). The major drawback of this approach is the risk of false-negative results due to clonal evolution. We investigated the stability of V delta 2D delta 3 rearrangements in a group of 56 childhood B-precursor ALL patients by PCR and Southern blot analysis. At the PCR level, V delta 2D delta 3-to-J alpha rearranged subclones (one pathway for secondary TCR delta recombination) were demonstrated in 85.2% of V delta 2D delta 3-positive patients tested, which showed that small subclones are present in the large majority of patients despite apparently monoclonal TCR delta Southern blot patterns. Sequence analysis of V delta 2D delta 3J alpha rearrangements showed a biased J alpha gene usage, with HAPO5 and J alpha F in 26 of 32 and 6 of 32 clones, respectively. Comparison of V delta 2D delta 3 rearrangement status between diagnosis and first relapse showed differences in seven of eight patients studied. In contrast, from first relapse onward, no clonal changes were observed in six patients studied. To investigate the occurrence of crosslineage TCR delta rearrangements in normal B and T cells, fluorescence-activated cell sorter-sorted peripheral blood CD19+/CD3- and CD19-/CD3+ cell populations from three healthy donors were analyzed. V delta 2D delta 3 rearrangements were detected at low frequencies in both B and T cells, which suggests that V delta 2-to-D delta 3 joining also occurs during normal B-cell differentiation. A model for crosslineage TCR delta rearrangements in B-precursor ALL is deduced that explains the observed clonal changes between diagnosis and relapse and is compatible with multistep leukemogenesis of B-precursor ALL. 相似文献
2.
Transition metal complexes containing vanadium IV have been shown to
modulate the cellular redox potential and catalyse the generation of
reactive oxygen intermediates (ROI). Since sperm function is exquisitely
susceptible to ROI, we examined the effects of stable chelate complexes of
vanadocenes on human sperm motility. We synthesized seven structurally
distinct chelate complexes of bis(cyclopentadienyl)vanadium(IV) with
bidentate ligands [i.e. vanadocene acetylacetonato monotriflate (VDacac),
vanadocene hexafluoro acetylacetonato monotriflate (VDHfacac), vanadocene
N-phenyl benzohydroxamato monotriflate (VDPH), vanadocene acethydroxamato
monotriflate (VDH), vanadocene catecholate (VDCAT), vanadocene bipyridino
ditriflate (VDBPY), and vanadocene dithiocarbamate monotriflate (VDDTC)],
and evaluated their spermicidal activity using computer-assisted sperm
analysis (CASA; Hamilton-Thorne). All seven chelate complexes of vanadocene
elicited potent spermicidal activity at micromolar concentrations (EC50
values: 3.9-106 microM) without affecting the sperm acrosome integrity. The
catecholate and acetylacetonate complexes of vanadocene were the most
active and the bipyridyl complex the least active with an order of efficacy
VDCAT > VDacac > VDDTC > VDPH > VDH > VDHfacac > VDBPY.
The spermicidal activity of chelate complexes of vanadocenes was rapid and
irreversible since the treated spermatozoa underwent apoptosis, as
determined by the flow cytometric analysis of mitochondrial membrane
potential, surface annexin V binding assay, in-situ nick-end labelling of
sperm nuclei, and confocal laser scanning microscopy. These results provide
unprecedented evidence that chelate complexes of vanadocene with bidentate
ligands have spermicidal and apoptosis inducing properties. These
vanadocene complexes, especially VDacac, may be useful as contraceptive
agents.
相似文献
3.
Production of immuno and biologically active erythropoietin was documented to occur in the human hepatoblastoma cell line HepG-2. The expression of the erythropoietin gene was further verified by Northern blot analysis using a single stranded RNA probe. In vitro studies showed that erythropoietin production by these cells was not stimulated by hypoxia or cobalt chloride, but was related to the proliferative activity of the cells in culture. In addition it was found that the secretion of erythropoietin was almost completely abrogated by tunicamycin, an inhibitor of N-linked glycosylation. This effect of tunicamycin was also observed in a permanently transfected cell line that secretes erythropoietin in large quantities. 相似文献
4.
Mice are a widely utilized in vivo model for translational salivary gland research but must be used with caution. Specifically, mouse salivary glands are similar in many ways to human salivary glands (i.e., in terms of their anatomy, histology, and physiology) and are both readily available and relatively easy and affordable to maintain. However, there are some significant differences between the two organisms, and by extension, the salivary glands derived from them must be taken into account for translational studies. The current review details pertinent similarities and differences between human and mouse salivary glands and offers practical guidelines for using both for research purposes. 相似文献
5.
Oral Diseases (2013) 19 , 236–244 Saliva plays a major role in maintaining oral health. Patients afflicted with a decrease in saliva secretion (symptomatically, xerostomia) exhibit difficulty in chewing and swallowing foods, tooth decay, periodontal disease, and microbial infections. Despite recent improvements in treating xerostomia (e.g., saliva stimulants, saliva substitutes, and gene therapy), there is a need of more scientific advancements that can be clinically applied toward restoration of compromised salivary gland function. Here we provide a summary of the current salivary cell models that have been used to advance restorative treatments via development of an artificial salivary gland. These models represent initial steps toward clinical and translational research, to facilitate creation of clinically safe salivary glands. Further studies in salivary cell lines and primary cells are necessary to improve survival rates, cell differentiation, and secretory function. Additionally, the characterization of salivary progenitor and stem cell markers are necessary. Although these models are not fully characterized, their improvement may lead to the construction of an artificial salivary gland that is in high demand for improving the quality of life of many patients suffering from salivary secretory dysfunction. 相似文献
6.
To define further the role of hemin-controlled repressor (HCR) in globin synthesis, we studied its effect on the synthesis of individual globin chains in a rabbit reticulocyte lysate cell-free system. In the presence of HCR there was a marked globin chain imbalance, resulting in a lowered alpha/beta ratio. These findings in vitro may have relevance to certain clinical heme deficiency states in which a similar globin chain imbalance has been observed. 相似文献
7.
8.
The human cell line PER.C6 provides a new manufacturing system for the production of influenza vaccines 总被引:5,自引:0,他引:5
Influenza viruses for vaccine production are currently grown on embryonated eggs. This manufacturing system conveys many major drawbacks such as inflexibility, cumbersome down stream processing, inability of some strains to replicate on eggs to high enough yields, and selection of receptor-binding variants with reduced antigenicity. These limitations emphasize the need for a cell line-based production system that could replace eggs in the production of influenza virus vaccines in a pandemic proof fashion. Here we present the efficient propagation of influenza A and B viruses on the fully characterized and standardized human cell line PER.C6. 相似文献
9.
W. Koudstaal L. Hartgroves M. Havenga I. Legastelois C. Ophorst M. Sieuwerts D. Zuijdgeest R. Vogels J. Custers E. de Boer-Luijtze O. de Leeuw L. Cornelissen J. Goudsmit W. Barclay 《Vaccine》2009
Reverse genetics, the generation of influenza viruses from cDNA, presents a rapid method for creating vaccine strains. The technique necessitates the use of cultured cells. Due to technical and regulatory requirements, the choice of cell lines for production of human influenza vaccines is limited. PER.C6® cells, among the most extensively characterized and documented cells, support growth of all influenza viruses tested to date, and can be grown to high densities in large bioreactors in the absence of serum or micro carriers. Here, the suitability of these cells for the generation of influenza viruses by reverse genetics was investigated. A range of viruses reflective of vaccine strains was rescued exclusively using PER.C6 cells by various transfection methods, including an animal component-free procedure. Furthermore, a whole inactivated vaccine carrying the HA and NA segments of A/HK/156/97 (H5N1) that was both rescued from and propagated on PER.C6 cells, conferred protection in a mouse model. Thus PER.C6 cells provide an attractive platform for generation of influenza vaccine strains via reverse genetics. 相似文献
10.
Koldijk MH Bogaards JA Kostense S de Vocht M Gijsbers L Ter Haak M Ophorst C Brakenhoff JP Weverling GJ Guichoux JY Uytdehaag F Lewis J Goudsmit J Marzio G 《Vaccine》2007,25(39-40):6872-6881
Ensuring complete viral inactivation is critical for the safety of vaccines based on an inactivated virus. Detection of residual infectious virus is dependent on sensitivity of the assay, sample volume analyzed and the absence of interference with viral infection. Here we describe the development and qualification of a sensitive cell-based assay for the detection of residual infectious West Nile Virus (WNV). The results of the assay are in good agreement with the assumption that at low concentrations the number of infectious units in relatively small samples follows a Poisson distribution. The assay can detect 1 infectious unit with a confidence of 99%, provides statistical controls for interference and can easily be scaled up to test large amounts of vaccine material. Furthermore, we show equivalence in sensitivity between the cell-based assay and an in vivo assay for detection of infectious WNV. Finally, the assay has been used for successful release testing of clinical lots of inactivated WNV vaccine. Given the principle and generic setup of the method we envision broad applicability to the detection of very low concentrations of infectious virus. 相似文献