Human Cytomegalovirus Can Induce Alteration of β-Actin mRNA and Microfilaments as Well as Induce Apoptosis in Mesenchymal Stem Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4261-4261
Author(s):  
Guoqing Wei ◽  
Maofang Lin ◽  
Zhen Cai ◽  
He Huang
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Mesenchymal Stem Cells ◽  
Electron Microscope ◽  
Human Cytomegalovirus ◽  
Transmission Electron Microscope ◽  
Cmv Infection ◽  
Transmission Electron ◽  
Stem Cells Transplantation ◽  
Actin Mrna

Abstract Mesenchymal stem cells (MSCs) is a very important cell system in bone marrow stromal and human cytomegalovirus (CMV) infection is one of the most common complications following stem cells transplantation. Our studies were to investigate the cytoskeleton change as well as the apoptosis induction effects of CMV infection on MSCs. In our study, MSCs were infected by 100,10,1 TCID50 of CMV. After 48h of culture with DMEM supplemented with 10% (v/v) fetal calf serum, MSCs shapes were observed with microscope. CMV particles and ultra structure in the cells were detected with transmission electron microscope. RT-PCR assay was used to detect the mRNA expression of CMV immediate early (IE) gene,β-actin gene and GAPDH genes. Flow Cytometry was used to detect the apoptotic cells. After the infection of CMV, shape of MSCs changed, turning from thin shuttle shape to round and thick ball shape, even escaping from wall. Cell shape changed more dominantly as the CMV titer increased. With transmission electron microscope, CMV particles and typical cellular apoptotic character as well as the ruptured- Microfilaments could be seen in the cells infected by CMV. MSCs infected by CMV could express IE mRNA and the expression of β-actin mRNA decreased in a time- and titer- dependent manner compared with the uninfected cells whose expression of GAPDH mRNA did not change much. With Flow Cytometry, it was found there were more apoptotic and necrosis cells in the cells infected by CMV. These results indicate that CMV can not only infect MSCs and destroy the cytoskeleton in the cells but also can induce apoptosis of the cells, much more attention of CMV infection on MSCs should be paid after stem cells transplantation.

Human Cytomegalovirus Can Activate and Induce Apoptosis in Mesenchymal Stem Cells.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4947-4947
Author(s):  
Guoqing Wei ◽  
Maofang Lin ◽  
He Huang ◽  
Zhen Cai
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Electron Microscope ◽  
Human Cytomegalovirus ◽  
Transmission Electron Microscope ◽  
Calf Serum ◽  
Cell System ◽  
Cmv Infection ◽  
Gapdh Gene ◽  
Transmission Electron

Abstract Objective Mesenchymal stem cells (MSCs) is a very important cell system in bone marrow stromal and human cytomegalovirus (CMV) infection is one of the most common complications following stem cells transplantation. Our studies were to investigate the CMV infection of MSCs as well as the apoptosis induction effects. Methods MSCs were infected by 100,10,1 TCID50 of CMV. After 48h of culture with DMEM supplemented with 10% (v/v) fetal calf serum, MSCs shapes were observed. CMV particles and ultra structure in the cells were detected with transmission electron microscope. RT-PCR assay was used to detect the mRNA expression of CMV immediate early (IE) gene and GAPDH gene. Flow Cytometry was used to detect the apoptotic cells. Results After the infection of CMV, shape of MSCs changed, turning from thin shuttle shape to round and thick ball shape, even escaping from wall. Cell shape changed more dominantly as the CMV titer increased. With transmission electron microscope, CMV particles and typical cellular apoptotic character could be seen in the cells. MSCs infected by CMV could express IE mRNA compare with the uninfected and there were more apoptotic and necrosis cells in the cells infected by CMV. Conclusion CMV can not only infect MSCs and activate in the cells but also can induce apoptosis of the cells.

NF-κB Inhibitor Pyrrolidinedithiocarbamate Can Increase the Infection and Activation of Cytomegalovirus in Mesenchymal Stem Cells.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4948-4948
Author(s):  
Guoqing Wei ◽  
Maofang Lin ◽  
He Huang ◽  
Zhen Cai
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Mesenchymal Stem Cells ◽  
Calf Serum ◽  
Close Relation ◽  
Clinical Work ◽  
Rt Pcr ◽  
Cmv Infection ◽  
Pcr Assay ◽  
Pp65 Antigen

Abstract Objective Because of the central role of the transcription factor nuclear factor-κB (NF-κB) in cell survival and proliferation in many kinds of cancer cells, NF-κB inhibitor may have a potential role in the therapy of cancer. Human cytomegalovirus (CMV) infection is one of the most common complications following stem cells transplantation. Some data report that CMV infection has a very close relation with NF-κB activation. But it is unknown what effect of NF-κB inhibitor pyrrolidinedithiocarbamate (PTDC) on the infection and activation of cytomegalovirus in mesenchymal stem cells(MSCs). Methods MSCs were infected by 1 TCID50 of CMV combined with/without 1μmol/L of PTDC. After 48h of culture with DMEM supplemented with 10% (v/v) fetal calf serum, MSCs shapes were observed. RT-PCR assay was used to detect the mRNA expression of CMV immediate early (IE) gene and GAPDH gene. Flow Cytometry was used to detect the CMV PP65 antigen positive cells. Results Shape of some MSCs changed after the infection of 1 TCID50 of CMV. But in MSCs infected by 1 TCID50 of CMV combined with 1μmol/L of PTDC, Cell shape changed more dominantly, almost all cells changed from thin shuttle shape to round and thick ball shape. While in the cells treated only with PDTC, shape of the cells did not changed. RT-PCR assay showed that there was a very bright band of CMV IE mRNA in MSCs infected by 1 TCID50 of CMV combined with 1μmol/L of PTDC compared with the cells infected only with 1 TCID50 of CMV. More CMV pp65 antigen positive cells were found in MSCs infected by 1 TCID50 of CMV combined with 1μmol/L of PTDC with Flow Cytometry. Conclusion NF-κB activation may affect CMV infection of cells. NF-κB inhibitor PTDC can increase the infection and activation of CMV in MSCs and we should pay more attention to CMV infection when we use NF-κB inhibitor in clinical work..

A Reproducible Selective Stain for Cardiac Sarcoplasmic Reticulum

1974 ◽  
Vol 32 ◽  
pp. 214-215
Author(s):  
R. A. Waugh ◽  
J. R. Sommer
Keyword(s):  
Complex System ◽  
Electron Microscope ◽  
Sarcoplasmic Reticulum ◽  
Transmission Electron Microscope ◽  
Electron Microscopic ◽  
Cardiac Sarcoplasmic Reticulum ◽  
Transmission Electron

Cardiac sarcoplasmic reticulum (SR) is a complex system of intracellular tubules that, due to their small size and juxtaposition to such electron-dense structures as mitochondria and myofibrils, are often inconspicuous in conventionally prepared electron microscopic material. This study reports a method with which the SR is selectively “stained” which facilitates visualizationwith the transmission electron microscope.

Surface Structure of the Spores of Glugea Weissenbergi

1969 ◽  
Vol 27 ◽  
pp. 238-239
Author(s):  
Sanford H. Vernick ◽  
Anastasios Tousimis ◽  
Victor Sprague
Keyword(s):  
Electron Microscope ◽  
Surface Structure ◽  
Transmission Electron Microscope ◽  
Mature Spore ◽  
Spore Surface ◽  
Sectioned Material ◽  
Transmission Electron ◽  
Surface Detail

Recent electron microscope studies have greatly expanded our knowledge of the structure of the Microsporida, particularly of the developing and mature spore. Since these studies involved mainly sectioned material, they have revealed much internal detail of the spores but relatively little surface detail. This report concerns observations on the spore surface by means of the transmission electron microscope.

A New High Resolution Transmission Electron Microscope with Second-Zone Objective Lens

1969 ◽  
Vol 27 ◽  
pp. 176-177 ◽  
Author(s):  
H. Tochigi ◽  
H. Uchida ◽  
S. Shirai ◽  
K. Akashi ◽  
D. J. Evins ◽  
...  
Keyword(s):  
Electron Microscope ◽  
High Resolution ◽  
Transmission Electron Microscope ◽  
Objective Lens ◽  
High Excitation ◽  
Transmission Electron ◽  
New Instrument

A New High Excitation Objective Lens (Second-Zone Objective Lens) was discussed at Twenty-Sixth Annual EMSA Meeting. A new commercially available Transmission Electron Microscope incorporating this new lens has been completed.Major advantages of the new instrument allow an extremely small beam to be produced on the specimen plane which minimizes specimen beam damages, reduces contamination and drift.

X-ray microanalysis of thin specimens in the transmission electron microscope at voltages up to 1000 Kv.

1978 ◽  
Vol 36 (1) ◽  
pp. 540-541
Author(s):  
G. Cliff ◽  
M.J. Nasir ◽  
G.W. Lorimer ◽  
N. Ridley
Keyword(s):  
Electron Microscope ◽  
Transmission Electron Microscope ◽  
Weight Fraction ◽  
Present Series ◽  
Constant Independent ◽  
X Ray ◽  
Transmission Electron ◽  
Series Of Experiments ◽  
Image Position ◽  
X Ray Absorption

In a specimen which is transmission thin to 100 kV electrons - a sample in which X-ray absorption is so insignificant that it can be neglected and where fluorescence effects can generally be ignored (1,2) - a ratio of characteristic X-ray intensities, I1/I2 can be converted into a weight fraction ratio, C1/C2, using the equationwhere k12 is, at a given voltage, a constant independent of composition or thickness, k12 values can be determined experimentally from thin standards (3) or calculated (4,6). Both experimental and calculated k12 values have been obtained for K(11<Z>19),kα(Z>19) and some Lα radiation (3,6) at 100 kV. The object of the present series of experiments was to experimentally determine k12 values at voltages between 200 and 1000 kV and to compare these with calculated values.The experiments were carried out on an AEI-EM7 HVEM fitted with an energy dispersive X-ray detector.

Aspects of microanalysis in a transmission electron microscope

1978 ◽  
Vol 36 (1) ◽  
pp. 510-511
Author(s):  
R. Sinclair ◽  
B.E. Jacobson
Keyword(s):  
Grain Size ◽  
Electron Beam ◽  
Electron Microscope ◽  
Chemical Analysis ◽  
Transmission Electron Microscope ◽  
Vapor Deposition ◽  
Critical Currents ◽  
X Ray ◽  
Fine Grain ◽  
Transmission Electron

INTRODUCTIONThe prospect of performing chemical analysis of thin specimens at any desired level of resolution is particularly appealing to the materials scientist. Commercial TEM-based systems are now available which virtually provide this capability. The purpose of this contribution is to illustrate its application to problems which would have been intractable until recently, pointing out some current limitations.X-RAY ANALYSISIn an attempt to fabricate superconducting materials with high critical currents and temperature, thin Nb3Sn films have been prepared by electron beam vapor deposition [1]. Fine-grain size material is desirable which may be achieved by codeposition with small amounts of Al2O3 . Figure 1 shows the STEM microstructure, with large (∽ 200 Å dia) voids present at the grain boundaries. Higher quality TEM micrographs (e.g. fig. 2) reveal the presence of small voids within the grains which are absent in pure Nb3Sn prepared under identical conditions. The X-ray spectrum from large (∽ lμ dia) or small (∽100 Ǻ dia) areas within the grains indicates only small amounts of A1 (fig.3).

The direct determination of magnetic domain wall profiles using a split detector STEM

1978 ◽  
Vol 36 (1) ◽  
pp. 466-467
Author(s):  
J.N. Chapman ◽  
P.E. Batson ◽  
E.M. Waddell ◽  
R.P. Ferrier
Keyword(s):  
Electron Microscope ◽  
Domain Wall ◽  
Transmission Electron Microscope ◽  
Domain Walls ◽  
Photographic Plate ◽  
Magnetic Domain ◽  
Direct Determination ◽  
Quantitative Information ◽  
Scanning Transmission Electron Microscope ◽  
Transmission Electron

By far the most commonly used mode of Lorentz microscopy in the examination of ferromagnetic thin films is the Fresnel or defocus mode. Use of this mode in the conventional transmission electron microscope (CTEM) is straightforward and immediately reveals the existence of all domain walls present. However, if such quantitative information as the domain wall profile is required, the technique suffers from several disadvantages. These include the inability to directly observe fine image detail on the viewing screen because of the stringent illumination coherence requirements, the difficulty of accurately translating part of a photographic plate into quantitative electron intensity data, and, perhaps most severe, the difficulty of interpreting this data. One solution to the first-named problem is to use a CTEM equipped with a field emission gun (FEG) (Inoue, Harada and Yamamoto 1977) whilst a second is to use the equivalent mode of image formation in a scanning transmission electron microscope (STEM) (Chapman, Batson, Waddell, Ferrier and Craven 1977), a technique which largely overcomes the second-named problem as well.

A General Method for Spectrometer Design in the Scoff Approximation

1976 ◽  
Vol 34 ◽  
pp. 538-539
Author(s):  
J. R. Fields
Keyword(s):  
Electron Microscope ◽  
Transmission Electron Microscope ◽  
Energy Analysis ◽  
Incident Beam ◽  
Scanning Transmission Electron Microscope ◽  
Chemical Identification ◽  
Scanning Transmission Electron ◽  
Transmission Electron ◽  
Scanning Transmission ◽  
General Method

The energy analysis of electrons scattered by a specimen in a scanning transmission electron microscope can improve contrast as well as aid in chemical identification. In so far as energy analysis is useful, one would like to be able to design a spectrometer which is tailored to his particular needs. In our own case, we require a spectrometer which will accept a parallel incident beam and which will focus the electrons in both the median and perpendicular planes. In addition, since we intend to follow the spectrometer by a detector array rather than a single energy selecting slit, we need as great a dispersion as possible. Therefore, we would like to follow our spectrometer by a magnifying lens. Consequently, the line along which electrons of varying energy are dispersed must be normal to the direction of the central ray at the spectrometer exit.

Resolution in the Scanning Transmission Electron Microscope

1972 ◽  
Vol 30 ◽  
pp. 454-455
Author(s):  
M. G. R. Thomson
Keyword(s):  
Electron Microscope ◽  
Transmission Electron Microscope ◽  
Spherical Aberration ◽  
Signal To Noise Ratio ◽  
Scanning Transmission Electron Microscope ◽  
Dark Field ◽  
Objective Lens ◽  
Scanning Transmission Electron ◽  
Transmission Electron ◽  
Scanning Transmission

The variation of contrast and signal to noise ratio with change in detector solid angle in the high resolution scanning transmission electron microscope was discussed in an earlier paper. In that paper the conclusions were that the most favourable conditions for the imaging of isolated single heavy atoms were, using the notation in figure 1, either bright field phase contrast with β0⋍0.5 α0, or dark field with an annular detector subtending an angle between ao and effectively π/2.The microscope is represented simply by the model illustrated in figure 1, and the objective lens is characterised by its coefficient of spherical aberration Cs. All the results for the Scanning Transmission Electron Microscope (STEM) may with care be applied to the Conventional Electron Microscope (CEM). The object atom is represented as detailed in reference 2, except that ϕ(θ) is taken to be the constant ϕ(0) to simplify the integration. This is reasonable for θ ≤ 0.1 θ0, where 60 is the screening angle.

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