Unusual production of gross murine leukemia virus (GrMuLV) in murine lymphoblasts as studied by electron microscopy
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In view of the known differences in sensitivity to glucocorticoids among functional subcategories of lympocytes in the mouse and the awareness that different subcategories of acute lymphocytic leukemia may represent proliferation of different clones of lymphoblasts, we examined lymphoblast populations with known surface markers for differences in specific cytoplasmic glucocorticoid receptor activity. Lymphoblasts were divided into those that formed sheep erythrocyte rosettes (T-lymphoblasts) and those that lacked all surface markers (null lymphoblasts). The latter were further subdivided by their ability to stimulate allogeneic donors in mixed lymphocyte culture (MLC). Lack of stimulation in MLC is a characteristic of mature T-cells. Eighteen patients had null lymphoblasts that did stimulate in MLC, and glucocorticoid binding sites in these ranged from 4,096 to 21,869 sites/cell (median, 7,571). Eighteen patients with T-lymphoblasts had binding sites ranging from 0 to 5,887 sites/cell (median, 2,173). This difference in specific glucocorticoid receptor levels is highly significant ( p < 0.001). Nine patients whose lymphoblasts lacked identifiable surface markers but failed to stimulate in MLC had intermediate values for receptor sites. Thus it appears that null lymphoblasts are more likely to have higher numbers of specific glucocorticoid receptor than T-lymphoblasts. The differential capacity of lymphoblasts to bind steroid may prove useful in designing therapeutic regimens for the different subcategories of acute lymphocytic leukemia.
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We aim to detect differential expressions of LMP genes(LMP-1,LMP-2A,LMP-2B) in normal human lymphocytes and EBV-transformed lymphoblasts in vitro.Firstly,expression levels of LMP genes(LMP-1,LMP-2A,LMP-2B) in the two kinds of cells were detected by using real-time PCR.Then LMP-1 protein expression was detected by Western bolt in the normal lymphocytes and EBV-transformed lymphoblasts.It was found that LMP-1 mRNA expression was 863 times up-regulated in the transformed lymphoblasts,as compared with that in the normal lymphocytes(P 0.01).Consequently,the expression of LMP-1 protein in EBV-transformed lymphoblasts was higher than that in the normal lymphocytes.LMP-2A mRNA expression was 1763 times up-regulated in the transformed lymphoblasts as compared with,that in the normal lymphocytes(P 0.05);LMP-2B mRNA expression was 90078 times up-regulated in the transformed lymphoblasts as compared with that in the normal lymphocytes(P 0.05).All the result show that the expressions of LMP-1,LMP-2A and LMP-2B genes would be upward in EBV-transformed lymphoblasts.
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Lymphoblasts from children with acute lymphoblastic leukemia (ALL) or malignant lymphoblastic lymphoma were studied using surface markers characteristic of T and B lymphocytes. A B-cell marker, i.e. surface immunoglobulin, was absent in all cases studied. Fourteen of 22 children (64%) had lymphoblasts with one or both markers of T lymphocytes, i.e. receptors for sheep erythrocytes (E) and/or human T-lymphocyte antigen (HTLA) detectable using heterologous antithymocyte sera absorbed with B lymphocytes. In all instances, lymphoblasts which carried E receptors also carried HTLA. However, lymphoblasts in 6 cases carried HTLA but not E receptors. It is possible that ALL may often involve T lymphocytes which are early in differentiation (i.e. prior to development of E receptors) or, alternatively, that E receptors may be lost from T cells following malignant transformation. Thymus enlargement was found only in cases of ALL or lymphoma where T markers were present. Lymphoblasts carried the same markers when examined in various sites and at various times from the same patient.
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Summary Cell volumes of lymphoblasts from 75 cases of acute lymphocytic leukaemia (ALL) and lymphocytes from 33 normal individuals were determined using a Coulter Counter Model H4 Channelyzer. The average mean cell volume (MCV) and the model volume (MV) of lymphoblasts were larger than the MCV and MV of normal lymphocytes (P < 0.01). In addition, the cell volumes of lymphoblasts from patients with ALL were more heterogeneous than normal lymphocytes. When the volumes of lymphoblasts were compared to the FAB classification, lymphoblasts from cases of L3 were larger than those from L2 and the latter were larger than lymphoblasts from the L1 subtype. When the volumes of lymphoblasts were compared to an immunological classification, lymphoblasts from cases of B cell ALL were larger than those from non-B, non-T and T cell ALL. The volume of lymphoblasts, however, showed no significant predictive value in the determination of complete remission, duration of first remission, or survival.
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Acute lymphocytic leukemia
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Correlative light and electron microscopy (CLEM) is revolutionizing how cell samples are studied. CLEM provides a combination of the molecular and ultrastructural information about a cell. For the execution of CLEM experiments, multimodal fiducial landmarks are applied to precisely overlay light and electron microscopy images. Currently applied fiducials such as quantum dots and organic dye-labeled nanoparticles can be irreversibly quenched by electron beam exposure during electron microscopy. Generally, the sample is therefore investigated with a light microscope first and later with an electron microscope. A versatile fiducial landmark should offer to switch back from electron microscopy to light microscopy while preserving its fluorescent properties. Here, we evaluated green fluorescent and electron dense nanodiamonds for the execution of CLEM experiments and precisely correlated light microscopy and electron microscopy images. We demonstrated that green color emitting fluorescent nanodiamonds withstand electron beam exposure, harsh chemical treatments, heavy metal straining, and, importantly, their fluorescent properties remained intact for light microscopy.
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Precursor cell
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A rapid method is described for obtaining ultrathin sections from light microscopy sections. Five‐micrometre epoxy sections, heat‐flattened to slides, were affixed to the tips of plastic blocks by light‐curable dental bond, and cured while still on the microscope stage by illumination with blue light for 2 min. Sections were detached from the slides by rapid cooling and then resectioned for electron microscopy.
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Cryo-Electron Microscopy
Biological specimen
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This article presents an overview of microscopy and its ability to assist in understanding what happens in cells and tissues. From the 1960s to 1980s, electron microscopy was the best way to understand cell processes, but the advent in the mid-1980s of light microscopy and the ability to do fluorescence imaging displaced electron microscopy in this area. However, the 21st century has seen several improvements in electron microscopy that, along with the need for more detailed ultrastructural information, make it again very attractive in the study of cells, tissues, and organs, and electron microscopy has resumed its place as the preeminent method in understanding cell processes.
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