New methods to simultaneously localize proteoglycan by LM and EM

1978 ◽  
Vol 36 (2) ◽  
pp. 672-673
Author(s):  
Nona Shepard
Keyword(s):  
Articular Cartilage ◽  
Electron Micrographs ◽  
Toluidine Blue ◽  
Chemical Interaction ◽  
Ruthenium Red ◽  
Cationic Dyes ◽  
Keratan Sulphate ◽  
New Methods ◽  
Pathological Conditions ◽  
Sectioned Tissue

Proteoglycan is a major component ofa cartilage and up to 70% can be lost during routine processing unless special measures are taken. With this in mind and the fact that proteoglycan loss and or alteration is primary in most pathological conditions of articular cartilage, full retention is, therefore, essential to validate the interpreting of electron micrographs.The cationic dyes, Safranin 0 and Toluidine Blue 0, seemed like likely candidates as their chemical interaction with proteoglycan has been well documented by Rosenberg. Both dyes bind stoichlometrically to polyanions; one molecule of dye to each negatively charged chondroitin 6 - sulfate or keratan sulphate molecule. Szirmal has reported that cationic dyes bind proteoglycan of fresh sectioned tissue. Therefore, following Insitu fixation with Safaranin 0 or Toluidinz Blue 0, thz resulting Image would be a quantitative example of proteoglycan localization and if present in sufficent mass would scatter electrons. Ruthenium red is widely used for E. M., but we found it of little value as an L. M. stain

Pinocytotic-Like Forms of Mitochondria From Rat Anterior Pituitary

1968 ◽  
Vol 26 ◽  
pp. 146-147
Author(s):  
Burton B. Silver
Keyword(s):  
Electron Micrographs ◽  
Conformational Changes ◽  
Anterior Pituitary ◽  
Dynamic State ◽  
Surrounding Cell ◽  
The Matrix ◽  
The Moment ◽  
Cell Medium ◽  
Sectioned Tissue ◽  
State Of Activity

Sectioned tissue rarely indicates evidence of what is probably a highly dynamic state of activity in mitochondria which have been reported to undergo a variety of movements such as streaming, divisions and coalescence. Recently, mitochondria from the rat anterior pituitary have been fixed in a variety of configurations which suggest that conformational changes were occurring at the moment of fixation. Pinocytotic-like vacuoles which may be taking in or expelling materials from the surrounding cell medium, appear to be forming in some of the mitochondria. In some cases, pores extend into the matrix of the mitochondria. In other forms, the remains of what seems to be pinched off vacuoles are evident in the mitochondrial interior. Dense materials, resembling secretory droplets, appear at the junction of the pores and the cytoplasm. The droplets are similar to the secretory materials commonly identified in electron micrographs of the anterior pituitary.

Application of selected cationic dyes for the semiquantitative estimation of glycosaminoglycans in histological sections of articular cartilage by microspectrophotometry

1996 ◽  
Vol 28 (8) ◽  
pp. 577-590 ◽  
Author(s):  
K. Király ◽  
T. Lapveteläinen ◽  
J. Arokoski ◽  
K. Törrönen ◽  
L. Módis ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Cationic Dyes ◽  
Histological Sections

scholarly journals Age-related changes in the composition and structure of human articular-cartilage proteoglycans

1978 ◽  
Vol 176 (3) ◽  
pp. 683-693 ◽  
Author(s):  
M T Bayliss ◽  
S Y Ali
Keyword(s):  
Hyaluronic Acid ◽  
Articular Cartilage ◽  
Age Groups ◽  
Human Articular Cartilage ◽  
Human Cartilage ◽  
Keratan Sulphate ◽  
Age Related ◽  
Composition And Structure ◽  
Normal Human ◽  
Cartilage Proteoglycans

1. Analysis of the purified proteoglycans extracted from normal human articular cartilage with 4M-guanidinium chloride showed that there was an age-related increase in their content of protein and keratan sulphate. 2. The hydrodynamic size of the dissociated proteoglycans also decreased with advancing age, but there was little change in the proportion that could aggregate. 3. Results suggested that some extracts of aged-human cartilage had an increased content of hyaluronic acid compared with specimens from younger patients. 4. Dissociated proteoglycans, from cartilage of all age groups, bind to hyaluronic acid and form aggregates in direct proportion to the hyaluronic acid concentration. 5. Electrophoretic heterogeneity of the dissociated proteoglycans was demonstrated on polyacrylamide/agarose gels. The number of proteoglycan species observed was also dependent on the age of the patient.

scholarly journals Biosynthesis of proteoglycan in vitro by cartilage from human osteochondrophytic spurs

1982 ◽  
Vol 206 (2) ◽  
pp. 329-341 ◽  
Author(s):  
Charles J. Malemud ◽  
Victor M. Goldberg ◽  
Roland W. Moskowitz ◽  
Lee L. Getzy ◽  
Robert S. Papay ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Articular Cartilage ◽  
Deae Cellulose ◽  
Culture Conditions ◽  
Human Articular Cartilage ◽  
Guanidinium Chloride ◽  
Tissue Slices ◽  
Keratan Sulphate ◽  
Defined Culture

Proteoglycan biosynthesis by human osteochondrophytic spurs (osteophytes) obtained from osteoarthritic femoral heads at the time of surgical joint replacement was studied under defined culture conditions in vitro. Osteophytes were primarily present in two anatomic locations, marginal and epi-articular. Minced tissue slices were incubated in the presence of [35S]sulphate or [14C]glucosamine. Osteophytes incorporated both labelled precursors into proteoglycan, which was subsequently characterized by CsCl-isopycnic-density-gradient ultracentrifugation and chromatography on Sepharose CL-2B. The material extracted with 0.5m-guanidinium chloride showed 78.1% of [35S]sulphate in the A1 fraction after centrifugation. Only 23.0% of the [35S]sulphate in this A1 fraction was eluted in the void volume of Sepharose CL-2B under associative conditions. About 60–80% of the [35S]sulphate in the tissue 4m-guanidinium chloride extract was associated with monomeric proteoglycan (fraction D1). The average partition coefficient (Kav.) of the proteoglycan monomer on Sepharose CL-2B was 0.28–0.33. Approx. 12.4% of this monomer formed stable aggregates with high-molecular-weight hyaluronic acid in vitro. Sepharose CL-2B chromatography of fractions with lower buoyant densities (fractions D2–D4) demonstrated elution profiles on Sepharose CL-2B substantially different than that of fraction D1, indicative of the polydisperse nature of the newly synthesized proteoglycan. Analysis of the composition and chain size of the glycosaminoglycans showed the following: (1) preferential elution of both [35S]sulphate and [14C]glucosamine in the 0.5m-LiCl fraction on DEAE-cellulose; (2) the predominant sulphated glycosaminoglycan was chondroitin 6-sulphate (60–70%), with 9–11% keratan sulphate in the monomer proteoglycan; (3) Kav. values of 0.38 on Sephadex G-200 and 0.48 on Sepharose CL-6B were obtained with papain-digested and NaBH4-treated D1 monomer respectively. A comparison of the synthetic with endogenous glycosaminoglycans indicated similar types. These studies indicated that human osteophytes synthesized in vitro sulphated proteoglycans with some characteristics similar to those of mature human articular cartilage, notably in the size of their proteoglycan monomer and predominance of chondroitin 6-sulphate. They differed from articular cartilage primarily in the lack of substantial quantities of keratan sulphate and aggregation properties associated with monomer interaction with hyaluronic acid.

Malaria blood mass

1937 ◽  
Vol 33 (8) ◽  
pp. 948-953
Author(s):  
G. Y. Repin
Keyword(s):  
Human Body ◽  
Medical Knowledge ◽  
New Methods ◽  
Pathological Conditions

Along with the progress of medical knowledge, the discovery of new methods of research, both clinical and laboratory, the concepts of some pathological conditions of the human body are changing.

scholarly journals Fucose content of keratan sulphates from bovine articular cartilage

1991 ◽  
Vol 273 (2) ◽  
pp. 307-310 ◽  
Author(s):  
G H Tai ◽  
G M Brown ◽  
H G Morris ◽  
T N Huckerby ◽  
I A Nieduszynski
Keyword(s):  
Molecular Weight ◽  
Articular Cartilage ◽  
Molecular Size ◽  
Gel Permeation Chromatography ◽  
Repeat Sequence ◽  
Keratan Sulphate ◽  
Bovine Articular Cartilage ◽  
Gel Permeation ◽  
Fucose Content ◽  
Galactose Content

Alkaline-borohydride-reduced keratan sulphate chains were isolated from bovine articular cartilage (6-8-year-old animals). Nine keratan sulphate fractions of increasing molecular weight were prepared by gel-permeation chromatography on a calibrated column of TSK 30 XL. The samples were analysed for fucose and galactose contents (% by wt. of keratan sulphate) and fucose/galactose ratio. The fucose content increased with molecular size, but the galactose content remained constant. It was concluded that the alpha(1→3)-linked fucose [Thornton, Morris, Cockin, Huckerby, Nieduszynski, Carlstedt, Hardingham & Ratcliffe (1989) Biochem. J. 260, 277-282] was located within the poly-N-acetyl-lactosamine repeat sequence of articular-cartilage keratan sulphate.

Tegumental glands in the paragnaths of Palaemon serratus (Crustacea: Natantia)

1989 ◽  
Vol 69 (1) ◽  
pp. 53-63 ◽  
Author(s):  
C. G. Alexander
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Apparatus ◽  
Electron Micrographs ◽  
Rough Endoplasmic Reticulum ◽  
Central Region ◽  
Toluidine Blue ◽  
Binding Agent ◽  
Food Stimulus ◽  
Digestive Function

The three lobed paragnath of Palaemon serratus (Pennant) contains numerous rosette-type tegumental glands often arranged in clusters of up to ten glands. Each gland is made up of around ten cells most of which stain deeply with toluidine blue and whose contents have a reticulate appearance in electron micrographs. One, or occasionally two, cells stain faintly and have a fine granular appearance in electron micrographs. The central region of each gland contains the main drainage duct together with feeder canals from the vesicular storage areas. The actively secreting region of each cell is peripheral and typified by abundant rough endoplasmic reticulum and Golgi apparatus. There are nerves adjacent to the glands and their contents are probably discharged in response to a food stimulus. The structure and possible function of the glands are discussed in relation to those of other species The secretions could act as a lubricant and as a binding agent to aid in the ingestion of particulate food. There is no evidence for any direct digestive function.

scholarly journals Articular cartilage cultured with interleukin 1. Increased release of link protein, hyaluronate-binding region and other proteoglycan fragments

1986 ◽  
Vol 238 (2) ◽  
pp. 571-580 ◽  
Author(s):  
A Ratcliffe ◽  
J A Tyler ◽  
T E Hardingham
Keyword(s):  
Articular Cartilage ◽  
Interleukin 1 ◽  
Binding Region ◽  
Rapid Loss ◽  
Keratan Sulphate ◽  
Link Protein ◽  
The Matrix ◽  
The Media ◽  
Average Size

Pig articular cartilage was maintained in culture for 3 days with and without porcine interleukin 1. The proteoglycans remaining in the cartilage and those released into the medium were analysed by using radioimmunoassays for the hyaluronate-binding region, link protein and keratan sulphate. In interleukin 1-treated cultures after 3 days there was 38% release of total glycosaminoglycans into the medium, 18% release of binding region, 14% release of link protein and 20% release of keratan sulphate epitope, whereas in control cultures the proportions released were much less (16, 9, 10 and 7% respectively). Characterization of the proteoglycans in the media after 1.5 days and 3 days of culture showed that interleukin 1 promoted the release of proteoglycan of large average size and also the release of link protein and of low-Mr binding region which was unattached to proteoglycan. Both the link protein and binding region released were able to bind to exogenously added hyaluronate, whereas the proteoglycan in the medium was not. The proteoglycans extracted from cultured cartilage were similar to those from fresh cartilage: they contained a high proportion of aggregating proteoglycans and some low-Mr binding region. The proportion of this binding region extracted from the interleukin 1-treated cartilage was increased. The presence of interleukin 1 in the cultures therefore appeared to increase the rate of proteolytic degradation of proteoglycan in the matrix and to lead to a more rapid loss of intact binding region, of link protein and of large proteoglycan fragments into the medium.

scholarly journals Abnormal mast cell granules in the beige (Chédiak-Higashi syndrome) mouse.

1975 ◽  
Vol 23 (2) ◽  
pp. 117-122 ◽  
Author(s):  
E Y Chi ◽  
D Lagunoff
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Alcian Blue ◽  
Toluidine Blue ◽  
Ruthenium Red ◽  
Chediak Higashi Syndrome

Mast cells of beige (C57BL/6J) (bg-j/bg-j) mice were examined histochemically and ultrastructurally. Mast cell granules in the beige mice were markedly enlarged and irregular in shape. Granule contents stained uniformly with acidified toluidine blue, but with ruthenium red and Alcian Blue-safranin, two components were evident. The rims of the abnormal granules stained with ruthenium red and with Alcian Blue; the centers of the granules were clear with ruthenium red and stained with safranin. Mast cell granules thus represent another abnormal organelle in the Chédiak-Higashi syndrome.

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