Ultrastructural localization of proteoglycans in tissue using Cuprolinic Blue according to the critical electrolyte concentration method: Comparison with biochemical data from the literature

1987 ◽  
Vol 19 (9) ◽  
pp. 520-526 ◽  
Author(s):  
Toin H. M. S. M. Van Kuppevelt ◽  
Twan L. M. Rutten ◽  
Charles M. A. Kuyper
Keyword(s):  
Electrolyte Concentration ◽  
Method Comparison ◽  
Ultrastructural Localization ◽  
Biochemical Data ◽  
Concentration Method ◽  
Cuprolinic Blue

scholarly journals Discrimination of the nucleolus by a critical electrolyte concentration method.

1993 ◽  
Vol 26 (1) ◽  
pp. 1-3 ◽  
Author(s):  
MARIA LUIZA S. MELLO ◽  
BENEDICTO DE CAMPOS VIDAL ◽  
MARLY M. DANTAS ◽  
ANA L. P. MONTEIRO
Keyword(s):  
Electrolyte Concentration ◽  
Concentration Method

scholarly journals Dermatan sulphate-rich proteoglycan associates with rat tail-tendon collagen at the d band in the gap region

1981 ◽  
Vol 197 (1) ◽  
pp. 213-216 ◽  
Author(s):  
J E Scott ◽  
C R Orford
Keyword(s):  
Phosphotungstic Acid ◽  
Electrolyte Concentration ◽  
Proteinase Inhibitors ◽  
Uranyl Acetate ◽  
Dermatan Sulphate ◽  
Concentration Method ◽  
Collagen Molecules ◽  
Tail Tendon ◽  
Rat Tail ◽  
Tendon Collagen

Rat tail tendon was stained with a cationic phthalocyanin dye, Cupromeronic Blue, in a ‘critical-electrolyte-concentration’ method [Scott (1980) Biochem. J. 187, 887-891] specifically to demonstrate proteoglycan by electron microscopy. Hyaluronidase digestion in the presence of proteinase inhibitors corroborated the results. Collagen was stained with uranyl acetate and/or phosphotungstic acid to demonstrate the banding pattern a-e in the D period. Proteoglycan was distributed about the collagen fibrils in an orthogonal array, the transverse elements of which were located almost exclusively at the d band, in the gap zone. The proteoglycan may inhibit (1) fibril radial growth by accretion of collagen molecules or fibril fusion, through interference with cross-linking, and (2) calcification by occupying the holes in the gap region later to be filled with hydroxyapatite.

scholarly journals Presence of human noro- and adenoviruses in river and treated wastewater, a longitudinal study and method comparison

2011 ◽  
Vol 10 (1) ◽  
pp. 87-99 ◽  
Author(s):  
Leena Maunula ◽  
Kirsi Söderberg ◽  
Heli Vahtera ◽  
Veli-Pekka Vuorilehto ◽  
Carl-Henrik von Bonsdorff ◽  
...  
Keyword(s):  
River Water ◽  
Method Comparison ◽  
Average Concentration ◽  
Treated Wastewater ◽  
Virus Concentration ◽  
Reverse Transcription Pcr ◽  
Concentration Method ◽  
Charged Membrane ◽  
Causative Agents ◽  
One Year

Norovirus (NoV) is one of the most common causative agents of waterborne gastroenteritis outbreaks. The main objective of the study was to determine the presence of human NoVs in river water and in treated wastewater (TW) released into the river. During a one-year survey in 2007/2008, NoVs were detected in 30.8% of river samples (20/65), and 40.5% of TW samples (17/45) with a real-time reverse transcription-PCR assay. NoVs were present in the river water in the winter and spring, coinciding with the NoV epidemiological peak in the community and the presence of NoVs in TW. Later in 2009, the concentration method used, pre-filtration with a Waterra filter combined with filtration through a negatively charged membrane, was evaluated against glass wool filtration and freeze-drying for the detection of adenoviruses in river water. The virus amounts measured varied greatly depending on the virus concentration method. The continued monitoring in the spring of 2009 also revealed that the average concentration of noro- and adenoviruses in TW was 2.64 × 103 and 1.29 × 104 pcr units per mL, respectively. No correlation between the presence of viruses and Escherichia coli was found. These results may be useful for risk assessment studies.

scholarly journals Collagen–proteoglycan interactions. Localization of proteoglycans in tendon by electron microscopy

1980 ◽  
Vol 187 (3) ◽  
pp. 887-891 ◽  
Author(s):  
J E Scott
Keyword(s):  
Electron Microscopy ◽  
Electrolyte Concentration ◽  
Staining Procedure ◽  
Adult Rabbit ◽  
Adult Rat ◽  
Concentration Method ◽  
Extended Space ◽  
Tail Tendon ◽  
Filamentous Material ◽  
Rat Tail

Proteoglycan in foetal- and adult-rat tail tendon and adult-rabbit achilles tendon was stained for electron microscopy with a cationic phthalocyanin-like dye, based on cinchomeronic acid, in a ‘critical electrolyte concentration’ method [Scott (1973) Biochem. Soc. Trans. 1, 787-806). Provided that the tissue was fixed with glutaraldehyde or formaldehyde, regular orthogonal perifibrillar arrays of filamentous material (proteoglycan) were observed, but no intra-fibrillar proteoglycan was seen. Specific proteoglycan-collagen interactions are inferred, and a model is proposed. Without fixation, the filamentous arrays disaggregated in the MgCl2 solutions (0.3 M) used during staining. End-to-end proteoglycan aggregation is implied. Tendon and cartilage are compared. Problems of electron-histochemical localization of extended space-filling polyanions by the use of cationic electron-dense precipitants are discussed, particularly polyanion-domain collapse, specificity of staining and fixation. A two-stage staining procedure that markedly enhances contrast is described, based on the multivalent nature of the dye, and the consequent anion-exchange properties of the dye-polyanion complex.

Apoptosis: identification by a critical electrolyte concentration method

APOPTOSIS ◽  
1996 ◽  
Vol 1 (3) ◽  
pp. 218-221 ◽  
Author(s):  
B. C. Vidal ◽  
L. F. Barbisan ◽  
S. S. Maria ◽  
J. Russo ◽  
M. L. S. Mello
Keyword(s):  
Electrolyte Concentration ◽  
Concentration Method

Ultrastructural localization of specific nucleoprotein fractions

1978 ◽  
Vol 36 (2) ◽  
pp. 204-205
Author(s):  
G. L. Brown
Keyword(s):  
Gel Electrophoresis ◽  
Mouse Liver ◽  
Polyacrylamide Gel Electrophoresis ◽  
Ultrastructural Localization ◽  
Polyacrylamide Gel ◽  
Acid Extraction ◽  
Acid Solutions ◽  
Low Salt ◽  
Liver Nuclei ◽  
Phosphate Groups

Bismuth (Bi) stains nucleoproteins (NPs) by interacting with available amino and primary phosphate groups. These two staining mechanisms are distinguishable by glutaraldehyde crosslinking (Fig. 1,2).Isolated mouse liver nuclei, extracted with salt and acid solutions, fixed in either formaldehyde (form.) or gl utaraldehyde (glut.) and stained with Bi, were viewed to determine the effect of the extractions on Bi stainina. Solubilized NPs were analyzed by SDS-polyacrylamide gel electrophoresis.Extraction with 0.14 M salt does not change the Bi staining characteristics (Fig. 3). 0.34 M salt reduces nucleolar (Nu) staining but has no effect on interchromatinic (IC) staining (Fig. 4). Proteins responsible for Nu and glut.- insensitive IC staining are removed when nuclei are extracted with 0.6 M salt (Fig. 5, 6). Low salt and acid extraction prevents Bi-Nu staining but has no effect on IC staining (Fig. 7). When nuclei are extracted with 0.6 M salt followed by low salt and acid, all Bi-staining components are removed (Fig. 8).

Ultrastructural Localization of Acid Mucosubstance in Skeletal Muscle

1967 ◽  
Vol 25 ◽  
pp. 52-53 ◽  
Author(s):  
William J. Dougherty ◽  
Samuel S. Spicer
Keyword(s):  
Striated Muscle ◽  
Divalent Cations ◽  
Ultrastructural Localization ◽  
Major Elements ◽  
Frozen Sections ◽  
Thorium Dioxide ◽  
Iron Solution ◽  
Coupling System ◽  
Psoas Muscles ◽  
Formalin Fixed

In recent years, considerable attention has focused on the morphological nature of the excitation-contraction coupling system of striated muscle. Since the study of Porter and Palade, it has become evident that the sarcoplastic reticulum (SR) and transverse tubules constitute the major elements of this system. The problem still exists, however, of determining the mechamisms by which the signal to interdigitate is presented to the thick and thin myofilaments. This problem appears to center on the movement of Ca++ions between myofilaments and SR. Recently, Philpott and Goldstein reported acid mucosubstance associated with the SR of fish branchial muscle using the colloidal thorium dioxide technique, and suggested that this material may serve to bind or release divalent cations such as Ca++. In the present study, Hale's iron solution adapted to electron microscopy was applied to formalin-fixed myofibrils isolated from glycerol-extracted rabbit psoas muscles and to frozen sections of formalin-fixed rat psoas muscles.

Immunoenzymatic demonstration of the simultaneous presence of transferrin, hemopexin and albumin in a same hepatocyte and their ultrastructural localization

1978 ◽  
Vol 36 (2) ◽  
pp. 88-89
Author(s):  
M. Kraemer ◽  
J. Foucrier ◽  
J. Vassy ◽  
M.T. Chalumeau
Keyword(s):  
Plasma Proteins ◽  
Rat Hepatocytes ◽  
Ultrastructural Localization ◽  
Carrier Proteins ◽  
Normal Cells ◽  
Adult Rat ◽  
Adult Rat Hepatocytes ◽  
Monospecific Antisera ◽  
Different Levels

Some authors using immunofluorescent techniques had already suggested that some hepatocytes are able to synthetize several plasma proteins. In vitro studies on normal cells or on cells issued of murine hepatomas raise the same conclusion. These works could be indications of an hepatocyte functionnal non-specialization, meanwhile the authors never give direct topographic proofs suitable with this hypothesis.The use of immunoenzymatic techniques after obtention of monospecific antisera had seemed to us useful to bring forward a better knowledge of this problem. We have studied three carrier proteins (transferrin = Tf, hemopexin = Hx, albumin = Alb) operating at different levels in iron metabolism by demonstrating and localizing the adult rat hepatocytes involved in their synthesis.Immunological, histological and ultrastructural methods have been described in a previous work.

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