| 11. |
- Tavecchio, Michele, et al.
(författare)
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Multi-parametric flow cytometric cell cycle analysis using TO-PRO-3 iodide (TP3): detailed protocols.
- 2008
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Ingår i: Acta histochemica. - : Elsevier BV. - 0065-1281. ; 110:3, s. 232-44
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Tidskriftsartikel (refereegranskat)abstract
- TO-PRO-3 iodide (TP3), a monomeric cyanine nucleic acid stain with a peak absorbance at 642 nm and emission at 661 nm, is best excited by a helium-neon (HeNe) laser (633nm). It was tested in monocytes and different cell lines under conditions of different fixatives, dye concentrations, labeling kinetics and RNAse concentrations for mono-, bi- and tri-parametric flow cytometric cell cycle analysis to establish the best protocol for DNA analysis in terms of G1 peak CV, G2/G1 ratio and minimal amount of debris. A linear increase in G1 peak position was found from 0.1 to 2 microM TP3 concentrations. Fixatives 70% ethanol or 1% methanol-free formaldehyde, followed by 70% ethanol, resulted in the best DNA histograms. Although different protocols were found to be cell-type specific, in general, excellent results were obtained with 30 min incubation with 0.5 microM TP3 plus RNAse in almost all cell lines tested. These data show that TP3 is an alternative method to propidium iodide (PI), the most commonly used DNA-specific probe in flow cytometry. The most important advantage of using TP3 in combination with other fluorochromes, such as fluorescein isothiocyanate (FITC) or phycoerythrin (PE) in bi- or tri-parametric flow cytometric analysis, is that there is no need for fluorescence compensation for the TP3 signals.
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| 12. |
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| 13. |
- Wu, CP, et al.
(författare)
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Immunohistochemical localization of programmed death-1 ligand-1 (PD-L1) in gastric carcinoma and its clinical significance
- 2006
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Ingår i: Acta Histochemica. - : Elsevier BV. - 0065-1281. ; 108:1, s. 19-24
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Tidskriftsartikel (refereegranskat)abstract
- The present study examined programmed death-1 ligand-1 (PD-L1) detected by immunohistochemical. labeling in 102 cases of human gastric carcinoma, 10 adenoma and 10 normal tissues. The relationship between PD-L1 immunolocalization and clinical pathological features, as well as the prognosis of gastric carcinoma, was explored. There was no PD-L1 detectable in normal gastric tissues and very weak immunolabeling in gastric adenomas, but it could be detected in 42.2% of gastric carcinoma tissues. There was no correlation between PD-L1 immunolocalization and patient age, sex, tumor location or the degree of tumor differentiation in the gastric carcinomas. However, PD-L1 immunodetection was significantly correlated to tumor size, invasion, lymph node metastasis and survival time of patients. PD-L1 immunolabeling was significantly enhanced (P<0.01) when the tumor infiltrated into the deep muscular layers, with lymph node metastasis or survival time of less than 2 years, Moreover, multivariate analysis demonstrated that PD-L1 immunodetection could be used as an independent factor to evaluate the prognosis of gastric carcinoma. (C) 2006 Elsevier GmbH. All rights reserved.
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| 14. |
- Ye, Q, et al.
(författare)
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A novel pattern of pp65-positive cytomegalic endothelial cells circulating in peripheral blood from a renal transplant recipient
- 2004
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Ingår i: Acta Histochemica. - : Elsevier BV. - 0065-1281. ; 106:2, s. 107-110
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Tidskriftsartikel (refereegranskat)abstract
- The present study reports a novel pattern of cytomegalic endothelial cells (CEC) in peripheral blood from a female renal transplant recipient infected with human cytomegalovirus (HCMV), which has not been reported previously. Localization of specific early antigen of HCMV, pp65 antigen, was examined by immunohistochemistry. Staining of an endothelial cell marker (CD34) was used to characterize endothelial cells. It is demonstrated that many leukocytes surrounded and adhered to a protein-like material, in which pp65-positive CEC were detected. The composition and function of this protein-like material are yet unknown. The patient tacked clinical symptoms of HCMV disease. Furthermore, similar localization patterns were found in other renal. transplant recipients suffering from HCMV infections as determined by real-time PCR to detect HCMV DNA in blood. These patients showed no or only minor clinical symptoms of HCMV infection. It is suggested that these novel Localization patterns of CEC may play a role in the host defense in patients infected with HCMV, but the exact relation between HCMV infection and CEC formation needs further investigation. (C) 2004 Elsevier GmbH. All rights reserved.
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| 15. |
- Zhang, XY, et al.
(författare)
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Expression pattern of apolipoprotein M during mouse and human embryogenesis
- 2004
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Ingår i: Acta Histochemica. - : Elsevier BV. - 0065-1281. ; 106:2, s. 123-128
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Tidskriftsartikel (refereegranskat)abstract
- Apolipoprotein M (apoM) is a recently discovered human apolipoprotein predominantly present in high-density lipoprotein (HDL), and in minor proportions in triglyceride-rich lipoprotein (TGRLP) and low-density lipoprotein (LDL). The gene encoding apoM is present in all mammalian genomes. The identity of the apoM gene of human, rat and mouse is over 80%. However, the (patho)physiological functions of apoM are unknown yet. In the present study, we investigated apoM expression patterns during mouse and human embryogenesis. ApoM transcripts were detectable in mouse embryos from day 7.5 to day 18.5. ApoM was expressed at low levels at day 7.5, its expression increased significantly at day 9.7, decreased at day 10.5, and then increased continually up to day 18.5. ApoM-positive cells appeared mainly in liver of day-12 embryos as detected by in situ hybridization. In day-15 embryos, apoM was expressed in both liver and kidney. During human embryogenesis, apoM was mainly expressed in liver and kidney and little was found in small intestine as determined by mRNA array of human fetal. normal tissues. ApoM was also detected in stomach and skeletal muscle in early stages of embryogenesis (3-5 months). (C) 2004 Elsevier GmbH. All rights reserved.
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