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At least 433 records · Page 24Linked to original sources
PubMed1996

Density shift of endocytic vesicles induced by uptake of colloidal gold particles in rat hepatocytes.

The aim of this study was to demonstrate the influence of endocytosed asialoorosomucoid gold complexes (ASOR-Au10, 10 = diameter of gold particles) on the density of endocytic compartments in rat hepatocytes when fractionated on linear sucrose gradients. We show that by loading hepatocytes with ASOR-Au10 for different time intervals, organelles containing gold particles are denser than organelles containing unconjugated asialoorosomucoid (ASOR). This density shift was observed in endosomes, which were loaded with a short pulse (15 minutes) of ASOR- Au10. Also lysosomes, loaded with ASOR-Au10 for 120 minutes at 37 degrees C, display a similar density shift.

Animals↗
PubMed1987

Receptor-mediated binding, endocytosis and cellular processing of macromolecules conjugated with colloidal gold.

Receptor-mediated expression of cellular functions assures biological specificity, regulation, and control of nutritive and metabolic requirements. The study of receptor-ligand interactions is a central theme in many published reports employing colloidal gold labeling. With the ligand directly conjugated with colloidal gold, the investigator is afforded the opportunity to observe all phases of cellular binding, endocytosis, lysosomal delivery and catabolism. Reviewed are published studies employing direct ligand conjugation with colloidal gold, the relative merits and disadvantages of this type of procedure and data from recent studies investigating endothelial receptor binding of proteins in the coagulation and fibrinolysis cascades.

Animals↗
PubMed1987

Radioactive colloidal gold as a tool to quantify extravasation of macromolecules in the rat cremaster muscle.

Since previously described simple methods were too insensitive to quantify the extent of microvascular permeability increase in the rat cremaster muscle, we applied a new quantitative technique using radioactive colloidal gold (198Au) as the macromolecular tracer. Dose-response effects on microvascular permeability were found with increasing doses of subscrotally injected serotonin or histamine. The presence of colloidal gold particles in the subendothelial space of the postcapillary venules, as verified ultrastructurally, indicated endothelial permeation of the tracer macromolecules. The increased permeability elicited by serotonin was inhibited in a dose-dependent way by serotonin receptor antagonists (potency: methysergide greater than or equal to ketanserin = ritanserin greater than cinanserin), suggesting the presence of functional S2-serotonergic receptors on postcapillary venular endothelial cells. A dose-related inhibition of the histamine-induced extravasation was seen with the H1 antagonists astemizole and azatidine (astemizole greater than azatidine); the H2 antagonists cimetidine and ranitidine had no inhibitory effect. The radioactive colloidal gold technique is a sensitive and reliable tool for pharmacological experiments aimed at investigating the extravasation phenomenon in small tissue samples such as the rat cremaster muscle.

Animals↗
PubMedDate not supplied

[Use of colloid gold conjugates for identification of actins of various origin].

Some optimized methods of purifying actin from animal, plant, and bacterial cells were developed. Variants of the preparation of antiactin antibodies are described which use both traditional methods of immunization and phase display technology and antigen adsorption on colloidal gold particles. The conjugates of colloidal gold with phalloidin and heavy meromyosin as well as with antibodies were proposed. It was shown that these markers make it possible to reliably identify actins of different origin by the methods of light, and electron microscopy and dot-analysis.

Actins↗
PubMed1987

A model of protein-colloidal gold interactions.

We prepared homogeneous populations of colloidal gold particles of various sizes. These were analyzed for size distribution and number of particles per unit volume. On exposure to increasing concentrations of insulin, myoglobin, protein A, peroxidase, serum albumin, galactosylated serum albumin, lactoferrin, transferrin, catalase, low-density lipoprotein, ferritin, and polymeric IgA, protein binding was a saturable process. Using serum albumin, we verified that a reversible equilibrium was reached within 15 minutes. Scatchard analysis of the interactions between all of these proteins and the gold particles resulted in a single component, linear relation. For a given particle size, the number of binding sites for various proteins was inversely proportional to their molecular weight. Conversely, when the size of particles was varied, the number of binding sites was directly proportional to the average area of each gold particle. All results are compatible with a monomolecular shell of protein surrounding the particle at saturation, the binding capacity being inversely proportional to the projection area of the protein. We present direct morphological evidence for this model. The affinity of the various proteins for the colloid also increased with molecular weight, and was not related to the protein isoelectric point. For globular proteins, the monomolecular shell model makes possible prediction of the number of molecules that will saturate a gold particle, if the average diameter of the gold particles and the molecular weight of the protein are known.

Adsorption↗
PubMed1988

The use of colloidal gold immunoelectron microscopy to diagnose varicella-zoster virus (VZV) infections by rapid discrimination between VZV, HSV-1 and HSV-2.

Colloidal gold immunoelectron microscopy was used to diagnose rapidly 53 cases clinically suspected of varicella-zoster virus (VZV) infection and one special case selected from another study on typical herpes simplex virus (HSV) infections. The viruses were identified and subsequently typed within 2.5 h by a direct labelling test for VZV, and within 3.5 h by an indirect labelling test with monoclonal antibodies against HSV type 1 and type 2. The protein A purified IgG fraction of human anti-VZV immunoglobulins was adsorbed to colloidal gold particles, and the specificity of the gold-labelled antibodies was tested with several human and animal herpesviruses. Viral envelopes did not crossreact in the direct labelling test. However, an indirect labelling procedure revealed that a small fraction of the anti-VZV antibodies crossreacted with the cores of herpes simplex virus and pseudorabies virus (Aujeszky disease virus). Virus-infected cellular material taken from typical herpetic lesions was used directly without virus propagation for virus typing. All cases (N = 54) were analyzed without knowing the clinical description of the results of cytopathologic examination (Tzanck smear) and viral culture. Forty-four cases were identified as VZV; however, 5 of the supposed VZV infections were proved to be HSV infections. Although the viral culture of the one HSV case was HSV- and VZV-negative, colloidal gold labelling identified the case as VZV infection. In 16 cases virus immunoglobulin complexes were detected by using gold-tagged antibodies against human immunoglobulins. Immunoglobulins on the viral envelopes did not interfere with virus typing by immunogold labelling.

Antibodies, Viral↗