The use of a carbodiimide-containing fixative for the immunohistochemical demonstration of coagulation factor VIII in rat vascular tissue

Pelt-Verkuil, E. van; Emeis, J.J.

doi:10.1007/bf00507823

Cited by 14 publications

(3 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Rand et al (20) showed dense FVIIIR:Ag precipitates concentrated in the basement membrane in umbilical vessels. However, Pelt-Verkuil et al (18) reported they did not detect FVIIIR:Ag in the subendothelium of intact arteries or de-endothelialized rat carotid artery using similar immunohistochemical methods. Our data localized FVIIIR:Ag to the basement membrane.…”

Section: Discussionmentioning

confidence: 99%

Immunocytochemical localization of factor VIII-related antigen in tumors of the human central nervous system

Miyagami¹,

Smith²,

McKeever³

et al. 1987

J Neuro-Oncol

View full text Add to dashboard Cite

Using both immunohisto- and immunocytochemical techniques with periodate-lysine-paraformaldehyde (PLP) fixation, we have studied the distribution of Factor VIII-related antigen (FVIIIR:Ag) in 12 cases of tumors of the human central nervous system (CNS) and one sample of non-tumor brain tissue. FVIIIR:Ag was found both extracellularly and intracellularly. It was localized in the vascular lumen, between endothelial cells, and in the endothelial cell basement membrane. In the endothelial cell cytoplasm, FVIIIR:Ag was found in the endoplasmic reticulum, perinuclear space, and in intracytoplasmic vacuoles and vesicles. Characteristic of malignant tumors (six out of seven) was a strongly-positive dilated endoplasmic reticulum. This may reflect increased FVIIIR:Ag synthesis in the endothelial cells of malignant tumors. Only one of five benign tumors showed such staining. Six of 12 tumors and the non-tumor brain showed perinuclear FVIIIR:Ag. Both ad- and abluminal vesicles in the tumor endothelial cells contained FVIIIR:Ag suggesting that endocytosis, transcellular transport, and/or endocytosis, as well as FVIIIR:Ag synthesis occurs. The non-tumor brain showed normal capillary structure and very little FVIIIR:Ag immunoreactivity. The relationship of these FVIIIR:Ag abnormalities to the hypercoagulable state seen in some malignant brain tumor patients remains to be clarified.

show abstract

Section: Discussionmentioning

confidence: 99%

Immunocytochemical localization of factor VIII-related antigen in tumors of the human central nervous system

Miyagami¹,

Smith²,

McKeever³

et al. 1987

J Neuro-Oncol

View full text Add to dashboard Cite

show abstract

“…We used rabbit antihuman F VIII R:Ag (Centraal Laboratorium Bloed-transfusiedienst, Amsterdam, The Netherlands) which was free of antifibronectin and antifibrinogen antibodies 16 at a 1:320 dilution. This antiserum yielded a single precipitation line against human cryoprecipitate in double immunodiffusion and immunoelectrophoresis.…”

Section: Immunohlstochemical Proceduresmentioning

confidence: 99%

“…The peroxidase conjugated antibody was used in the second step. 16 Smooth muscle cell (SMC) myosin (rabbit-antichicken gizzard myosin kindly provided by Professor Dr. Ute Groschel-Stewart, TH Darmstadt, West Germany 17 ) was demonstrated using a 1 :600 dilution of the primary antibody, followed by the FITC conjugated antibody in the second step.…”

Section: Immunohlstochemical Proceduresmentioning

confidence: 99%

Morphometry of air-drying-induced arteriosclerosis in rat carotid artery. Effect of air-flow rate.

Pelt-Verkuil¹,

Pelt²,

Jense³

1983

Arteriosclerosis

View full text Add to dashboard Cite

Arteriosclerosis in the Wistar rat carotid artery was induced by air drying of the endothelium, a procedure that caused media necrosis. We describe a number of technical modifications that facilitate the procedure and minimize damage to the vessel wall (i.e., media necrosis). A morphometric study of vessel wall changes induced by endothelial denudation at various air-flow rates showed that necrosis of the inner medial layer and lesion size and position were constant at all flow rates used (=s 28 ml/min). The extent of necrosis of the outer medial layers, the endothelial repair, and the shape of the lesions varied with the air-flow rate used. Since at an air-flow rate of 28 ml/min medial necrosis was minimal and myointimal lesion development was as extensive as at higher flow rates, we consider this air-flow rate to be optimal for this model of experimental arteriosclerosis. We hypothesize that the shape of the myointimal lesion is determined by the timespan of endothelial denudation and by the availability of smooth muscle cells in necrotic and normal media. Furthermore, the proliferation of the smooth muscle cells and their migration into the intima is probably dominant over the repopulation of the media by these cells. (Arteriosclerosis 3:441-451, September/October 1983) T he formation of arteriosclerotic, fibromuscularelastic intimal thickening following experimental endothelial denudation has been well documented in a number of animal models. 1 " 4 In rat arteries mechanical injury, 5 air-drying, 6 and balloon catheterization 78 have been shown to remove the endothelium and to cause myointimal thickening. Although these experiments were aimed at specificially causing endothelial injury, the underlying media was often affected as well.8 " 10 Fishman et al. 6 and Clowes et al. 11used the air-drying technique in the rat and claimed that injury, at least at the time of denudation, was confined to the endothelium. This apparent selectivity led us to choose the air-drying model for a study of the effect of experimental conditions on lesion development.In our study however, we found that this injury was not as selective as in other studies; 6 media necrosis (disappearance of smooth muscle cells) occurred in certain areas of the vessel wall. Furthermore, a different intimal thickening pattern was observed. Preliminary studies indicated that the velocity of infused air was important. To find the optimal conditions, we performed the air-drying method by lowering the airflow rate in steps of approximately 5 ml/min down to the lower limit of lesion induction. The air-drying technique of Fishman et al. 6 was modified, thereby facilitating surgery, avoiding mechanical injury at air inflow and outflow holes, and minimizing media necrosis.In this paper we shall present a univariate quantitative analysis of the changes in vessel wall morphology that we found when applying diminished air-flow rates to denude the artery. Special attention will be paid to parameters describing the resulting myointimal lesions, endothelial repai...

show abstract

The peroxidase antiperoxidase staining of factor VIII‐related antigen on cultured endothelial cells

Baughman

Herring

Glover

et al. 1984

J. Biomed. Mater. Res.

View full text Add to dashboard Cite

To prevent the premature occlusion of vascular prostheses, endothelium is being cultured experimentally onto synthetic flow surfaces. A rapid method of identifying cultured endothelium on the prosthesis is valuable for determining the degree of fibroblast and smooth muscle cell contamination and to screen for endothelial cell transformation. Fluorescent Factor VIII related antigen (FVIII-RA) staining has been used to identify cultured endothelium, but results in excessive staining of the underlying prosthesis, loss of morphologic detail, and deterioration of the FVIII-RA antibody reaction with time. We have applied the peroxidase antiperoxidase (PAP) method of antigen staining to permit staining of FVIII-RA and thereby to permit a sensitive and specific identification of human or canine endothelium with a concurrent analysis of morphologic detail.

show abstract

The use of a carbodiimide-containing fixative for the immunohistochemical demonstration of coagulation factor VIII in rat vascular tissue

Cited by 14 publications

References 21 publications

Immunocytochemical localization of factor VIII-related antigen in tumors of the human central nervous system

Immunocytochemical localization of factor VIII-related antigen in tumors of the human central nervous system

Morphometry of air-drying-induced arteriosclerosis in rat carotid artery. Effect of air-flow rate.

The peroxidase antiperoxidase staining of factor VIII‐related antigen on cultured endothelial cells

Contact Info

Product

Resources

About