scholarly journals HCMV pp65 antigenemia assay using indirect alkaline phosphatase staining method

1995 ◽  
Vol 16 (2) ◽  
pp. 125-129 ◽  
Author(s):  
TAKASHI KURIHARA ◽  
JUNKO HAYASHI ◽  
TOHRU MATSUOKA ◽  
AKIO ITO
Keyword(s):  
Alkaline Phosphatase ◽  
B Method ◽  
Alkaline Phosphatase Staining ◽  
Antigenemia Assay ◽  
Pp65 Antigenemia

scholarly journals Rapid Cytomegalovirus pp65 Antigenemia Assay by Direct Erythrocyte Lysis and Immunofluorescence Staining

1998 ◽  
Vol 36 (3) ◽  
pp. 638-640 ◽  
Author(s):  
Stephen K. N. Ho ◽  
Chi-Yuen Lo ◽  
Ignatius K. P. Cheng ◽  
Tak-Mao Chan
Keyword(s):  
Alkaline Phosphatase ◽  
Immunofluorescence Staining ◽  
Renal Transplant Recipients ◽  
Alkaline Phosphatase Staining ◽  
Erythrocyte Lysis ◽  
Indirect Immunofluorescence Staining ◽  
Antigenemia Assay ◽  
Immunosuppressed Patients ◽  
Pp65 Antigenemia ◽  
Assay Time

A rapid cytomegalovirus (CMV) pp65 antigenemia assay with direct erythrocyte lysis (DL) with 0.8% NH4Cl, followed by indirect immunofluorescence staining (IF), was evaluated with 82 blood samples from renal transplant recipients, and the results were compared to those of the conventional antigenemia assay with dextran sedimentation and two-cycle alkaline phosphatase, anti-alkaline phosphatase staining (DS-APAAP). The DL-IF modification gave a higher leukocyte yield compared to DS-APAAP (75.4 versus 54.9%;P < 0.05), with similar leukocyte viability rates of >95%. The DL-IF methodology involved fewer technical steps, and the assay time was shortened from 5 h to less than 3 h. Nineteen of the 82 samples concordantly tested positive for pp65 antigenemia by both assays, and the readings showed a good correlation (r = 0.996; P < 0.01). No discordant results were observed. We conclude that the CMV pp65 antigenemia assay by this novel DL-IF modification is technically simpler, cheaper, and less time-consuming but yields results comparable to those of the conventional DS-APAAP assay. The shortened assay time and increased capacity to handle more samples confer distinct advantages in the rapid diagnosis and prompt treatment of CMV disease in immunosuppressed patients.

Biocompatibility of Calcium Phosphate Ceramics Synthesized from Eggshell

2007 ◽  
Vol 330-332 ◽  
pp. 23-26 ◽  
Author(s):  
Nam Sik Oh ◽  
Yun Ho Na ◽  
S. W. Ji ◽  
S.W. Song ◽  
S.H. Oh ◽  
...  
Keyword(s):  
Stem Cells ◽  
Alkaline Phosphatase ◽  
Cell Attachment ◽  
Morphological Characteristics ◽  
Osteoblastic Differentiation ◽  
Human Bone ◽  
Pcr Analysis ◽  
Mixing Ratio ◽  
Culture Plate ◽  
Alkaline Phosphatase Staining

The aim of this paper was the HA and β-TCP powers were synthesized by a new wetchemical method using eggshell and phosphoric acid. The biocompatibility of synthesized natural HA, HA/β-TCP(50:50) and β-TCP derived from eggshell was compared with those of as commercial chemical powder with mesenchymal stem cells derived from human bone marrow. Development of crystalline phases of the mixtures was studied as functions of mixing ratio and temperature using X-ray diffractometer. The morphological characteristics of the calcined eggshell and synthesized powders were examined by scaning electron microscopy. The in-vitro cytotoxicity and cell attachment of sintered disks were examined using human bone marrowderived multipotent stem cells(hBMSCs). Cell response was characterized by MTT assay , Alkaline phosphatase stain and RT-PCR analysis. Pure HA was synthesized in the mixing ratio of 1:1.1 wt% at 900°C for 1h. the crystallization of HA was started at 800°C in the 1:1.1 mixing ratio, ant the HA phase was continued up to the high temperatures. In the ratio of 1:1.3 and 1:1.5 wt%, β-TCP was effectively synthesized at 900°C. In the 1:1.5 ratio, β-TCP phase was detected at 700°C, and complete crystallized β-TCP was observed above 900°C. At the higher temperature than 1000°C, the β-TCP was gradually decreased and α-TCP was observed. The HA and β-TCP disk does not exert cytotoxic effect on the hBMSCs undergoing osteoblastic differentiation. In addition, the hBMSCs are adhered on the surface of synthesized natural HA and β-TCP disk as successfully as on the culture plate or as commercial chemical HA and β-TCP disk. The hBMSCs adhered on either synthesized natural HA, β-TCP or as commercial chemical HA, β-TCP disk displays undistinguishable actin arrangement and cellular phenotypes, indicating that synthesized natural HA, β-TCP does not disrupt normal cellular responses. Analysis of differentiation of the hBMSCs cultured on culture plate, synthesized natural HA, β-TCP and as commercial chemichal HA, β-TCP disk shows that three matrices are able to support osteoblastic differentiation of the hBMSCs as accessed by alkaline phosphatase staining.

Massive cerebral fat embolism from hip surgery: a histologic study with alkaline phosphatase staining and thick celloidin sections

2000 ◽  
Vol 70 (5) ◽  
pp. 1793 ◽  
Author(s):  
William R Brown ◽  
Dixon M Moody ◽  
David A Stump ◽  
David M Colonna ◽  
Douglas J Kilgus
Keyword(s):  
Alkaline Phosphatase ◽  
Fat Embolism ◽  
Hip Surgery ◽  
Histologic Study ◽  
Alkaline Phosphatase Staining

Impaired Osteogenic Differentiation Of Mesenchymal Stem Cells Derived From Bone Marrow Of Patients With Low-Risk Myelodysplastic Syndromes

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 1579-1579
Author(s):  
Chunkang Chang ◽  
Chengming Fei ◽  
Youshan Zhao ◽  
Juan Guo ◽  
Xiao Li
Keyword(s):  
Alkaline Phosphatase ◽  
Osteogenic Differentiation ◽  
Lower Risk ◽  
Conflicts Of Interest ◽  
Osteoblastic Differentiation ◽  
Differentiation Markers ◽  
Alizarin Red ◽  
Hematopoietic Microenvironment ◽  
Alkaline Phosphatase Staining ◽  
Osteogenic Induction

Abstract Background The pathogenesis of MDS has not been completely understood, and insufficiency of the hematopoietic microenvironment can be an important factor. MSCs and osteoblasts are key components of the hematopoietic microenvironment. Studying osteoblastic differentiation of MSCs quantitatively may help to understand the pathogenesis of MDS. Methods 38 patients with MDS and 15 normal donors were investigated in this study. Osteoblastic differentiation assays were performed in 16 MDS cases and 8 controls. The expression of osteogenic differentiation markers were measured by real-time PCR. Alkaline phosphatase staining was performed with Alkaline Phosphatase staining kit after 3,7,14 days of incubation. ALP activity was assessed at 3, 7, and 10 days after osteogenic differentiation. Mineralization analysis was performed at 7, 14 and 21 days of osteogenic induction. The areas of mineralization were measured by Image-Pro Plus 6.0 software. Results Both MDS-MSCs and normal cells displayed same fibroblast-like morphology and similar antigen expression. The expression level of RUNX2 was significantly decreased in MSCs from MDS, compaired with normal controls, especially in lower-risk MDS. After osteogenic induction, lower-risk MDS showed lower alkaline phosphatase activity, less intense alizarin red S staining, and lower gene expression of osteogenic differentiation markers, however, higher-risk MDS was normal. Conclusions We concluded that impaired osteogenic differentiation of MSCs was seen mainly in patients with lower-risk MDS. It may contribute to the ineffective hamatopoiesis of MDS. Disclosures: No relevant conflicts of interest to declare.

Sign in / Sign up

Export Citation Format

Share Document