Ultrastructural histochemistry of the dento-epithelial junction. I. Phosphotungstic acid periodic acid-silver methenamine and periodic acid-thiosemicarbazide-silver proteinate.
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Animals↗
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Immunohistological characterization of mucin epitopes by pre-treatment of gastro-intestinal sections with periodic acid.
Periodate pretreatment of paraffin sections of ethanol-fixed gastrointestinal mucosae was used to characterize the carbohydrate or peptidic nature of mucin epitopes by immunoperoxidase. Immunoreactivity of monoclonal antibodies (MAbs) against histo-blood group related carbohydrate epitopes such as A, Lea, Lec, Sialosyl Lea, H type 2, I, T, Tn and sialosyl Tn dramatically decreased or even disappeared after periodate pretreatment of deparaffinized sections. In contrast, the immunoreactivity of MAbs against peptide mucin epitopes such as the gastric M1 mucin epitopes was almost unaffected by this treatment. Moreover, periodate treatment revealed cryptic peptide M1 mucin epitopes and the peptide MUSE11 epitope associated with the 20 amino acid tandem repeat (PDTRPAPGSTAPPAHGVTSA). An increase of cross-reactions of anti-human M1 MAbs with gastric epithelium of different vertebrate species was detected with periodate treatment. Our results suggest that this method can be useful for preliminary characterization of the biochemical nature of mucin epitopes (peptidic or saccharidic) and for demasking peptidic tumour markers which are hidden by saccharide molecules in normal tissues.
Adolescent↗
Enhancement of the periodic acid--Schiff (PAS) and periodic acid--thiocarbohydrazide--silver proteinate (PA-TCH-SP) reaction in LR white sections.
LR White is a well-suited resin for the demonstration of carbohydrates with the PAS or PA-TCH-SP reaction in semithin and ultrathin sections. The intensity of these reactions can be greatly enhanced by using 3 steps in tissue preparation, either singly or in combination: 1) The PAS reaction in semithin sections turns out stronger after partial (70% ethanol) than complete (100% ethanol) dehydration of the tissue before its transfer to 100% LR White. 2) Silver enhancement of the PA-TCH-SP reaction product can simply be effected by physical development of ultrathin sections (PA-TCH-SP-SE reaction). Least precipitates are formed in this procedure, when sections are mounted on uncoated gold grids, processed for cytochemistry, and thinly coated with carbon in the end. 3) The use of hot silver proteinate (50 degrees C) plus strong silver enhancement (15-20 min silver lactate developer) reveals minute concentrations of TCH-labelled aldehyde groups in the tissue that do not react with silver proteinate at room temperature.--Silver enhancement and the use of hot silver proteinate do not depend on LR White, but may also be applied to ultrathin sections of tissue embedded in other resins.
Acrylic Resins↗
Calcium is required for the mitogenic activation of lymphocytes by periodic acid oxidation.
This investigation of Ca2+ requirements for the mitogenic activation of lymphocytes by periodic acid has shown that oxidation by periodate causes an immediate and transient increase of Ca2+ influx and efflux in oxidized cells. Oxidized lymphocytes maintained in the medium containing 0.2 mM Ca2+ failed to proliferate or to produce IL-2, whereas a 1.4 mM Ca2+ concentration was shown to be sufficient to sustain cellular proliferation and IL-2 secretion. These results indicate that mitogenic activation of lymphocytes by periodic acid oxidation is Ca(2+)-dependent.
Animals↗
Indirect sulphonation of mucopolysaccharides and glycogen in tissue sections with the use of bisulphite or dithionite after periodic acid oxidation.
Tissue mucopolysaccharides and glycogen can be indirectly sulphated after being oxidized by periodic acid and treated with sodium bisulphite or dithionite in aqueous solution. The sulphated sites are darkly stained by Roluidine Blue and realted dyes at pH less than 1.0. The background is very pale or colourless. The stained sections resist extraction with 1% hydrochloric acid for 48 hr, but can be extracted by 5% ammonium hydroxide in ethanol in 1 hr. Other oxidizing agents cannot be substituted for periodic acid.
Dithionite↗
Periodic acid incubation can replace hydrochloric acid hydrolysis and trypsin digestion in immunogold--silver staining of bromodeoxyuridine incorporation in plastic sections and allows the PAS reaction.
We have examined the possibility of improving the present methods of detecting bromodeoxyuridine (BrdU) and for combining the PAS reaction with the BrdU detection by means of immunogold-silver staining (IGSS). This was done in testes fixed in Carnoy or Bouin, and in parts of the small intestine which were fixed in Carnoy or periodate-lysine-paraformaldehyde (PLP). All tissues were embedded in a mixture of glycol methacrylate and butanediol-monoacrylate. It was found to be impossible to carry out BrdU detection using HCl hydrolysis and trypsin digestion in combination with a PAS reaction. However, incubation of the plastic sections in periodic acid for a period of 30 minutes appeared to make it possible to eliminate the HCl denaturation step and to carry out a specific PAS reaction. Moreover, after incubation in periodic acid, trypsin digestion was no longer required to make the BrdU label accessible in GMA-embedded sections, nor to re-expose the antigenic sites in plastic sections of tissues fixed with cross-linking fixatives. In this way the loss of cell structures, which is inevitable when trypsin is used, can be avoided. Now a BrdU detection with improved morphology can be combined with the PAS reaction in the same plastic section in order to stain tissue carbohydrates. This is important for tumour diagnosis, where the PAS reaction can be very useful.
Animals↗
Oxidation of tissue polysaccharides by periodic acid in dimethyl sulfoxide and its anhydrous and aqueous mixtures.
Periodic acid (1% w/v) solvated by anhydrous dimethyl sulfoxide (DMSO) readily induced a strong Schiff reaction in a variety of structures containing polysaccharides, but not glycogen. With the increasing amounts of water added to DMSO, glycogen was also oxidized, while the selective localization of other polysaccharides remained unimpaired. Periodate, solvated in the anhydrous acetic acid-DMSO mixture, rapidly induced concomitant oxidation of nucin and glycogen-containing structures. Sodium bisulfite addition derivatives of carbonyls, induced by periodate oxidation in DMSO, were stained meta- and orthochromatically with toluidine blue at controlled pH. Certain metachromatic tissue components were strongly birefringent in polarized light in contrast to the identical structures oxidized by aqueous periodate. Marked differences in staining reactions elicited in identical structures by periodate in DMSO as compared with aqueous periodate suggest that DMSO-periodate method considerably enhances the range of histochemical oxidations by periodate.
Animals↗
[Detection of glomerular deposits of various renal diseases on light microscopy using periodic acid thionin [PATS]-chromotrope staining].
We devised a periodic acid thionine schiff (PATS)-chromotrope method to detect the glomerular deposits more distinctly than conventional staining methods. The PATS-chromotrope method was compared with other immunological staining methods, such as immunofluorescence method and avidin biotin complex method. Formalin-fixed, and paraffin-embedded renal tissues were obtained from 26 patients with IgA nephropathy, 8 with lupus nephritis, 4 with minimal change nephrotic syndrome, 3 with membrano-proliferative glomerulonephritis, and 3 with hepatitis-B associated nephropathy. Thionine Schiff reagent was used instead of fuchsin-schiff reagent to stain the basement membrane blue. Subsequently, chromotrope 2R was used to stain the glomerular deposits. For immunofluorescence method, frozen renal tissues were stained with FITC-labelled anti-human IgG, IgA, C3 and fibrinogen. For avidin biotin complex method, the same sections as PATS-chromotrope method were stained with anti-human IgG, IgA, and C3. In PATS-chromotrope method, deposits were identified in 9 of 26 specimens with IgA nephropathy, 3 of 8 specimens with lupus nephritis, and one of 3 specimens with hepatitis-B associated nephropathy. Localization of deposits in PATS-chromotrope method was identified more distinctly than immunofluorescence method or avidin biotin complex method. PATS-chromotrope method is useful to detect the deposition of immune complex on routine light microscopy in human glomerular disease.
Child↗
[Demonstration of an alkali PAS-effect using periodic acid at low concentration (author's transl)].
It has been shown that an isolated KOH PAS-effect of epithelial mucosubstances in the colonic mucosae of the rat can be demonstrated also without the conventionally used preliminary oxidation/reduction step. The method is based on the use of strongly diluted periodic acid after previous treatment of tissue sections with ethanolic KOH. As the positive reacting material has been proven to be sensitive against treatment with neuraminidase the modified KOH PAS-reaction should be related to the presence of acylated sialic acid residues in mucosubstances of the colonic mucosae of the rat.
Animals↗
[Application of "mild" periodic acid oxidation to the ultrahistochemical detection of sialic acid-containing compounds in human fallopian tube epithelium].
2 histochemical methods were used to analyse the carbohydrates in the human oviductal epithelium, namely the periodic acid-semicarbazide-silver proteinate and the 'mild' periodic acid-thiosemicarbazide-silver proteinate technique. It was possible to detect periodate reactive substances and sialic acid-containing compounds, namely separately. The 'mild' periodic acid oxidation is a selective technique for the histochemical demonstration of the existence of unsubstituted acylneuraminic acids. The ultrahistochemical results show that, firstly, periodate reactive particles are located in the cytoplasm of ciliated and nonciliated cells. The material appears in form of beta-granula (glycogen). Secondly, we could observe fine granular reactive products in the lysosomal-like and lipid droplet-like structures. In these subjects the positive granula encircling ring-like the homogeneous matrix occur in the peripheral zone of the morphological substrate investigated. By the way, these granula situated in the periphery of organelles are alone reacting after staining with 'mild' periodic acid-thiosemicarbazide-silver proteinate; this means, the glycogen particles are unstained. Thus the findings indicate that, in the cells of the human oviductal epithelium, sialic acid-containing compounds are ultrahistochemically visualized by 'mild' periodic acid oxidation in combination with thiosemicarbazide and silver proteinate.
Cilia↗
Thiosemicarbazide used after periodic acid makes methenamine silver staining of renal glomerular basement membranes faster and cleaner.
The periodic acid-methenamine silver staining technique, which is frequently used for demonstrating the renal glomerular basement membrane, requires a high degree of skill, and in some cases it may be difficult to obtain a good result. To overcome such difficulty and inconsistency, we have improved the method by performing methenamine silver staining after oxidation with periodic acid and subsequent application of thiosemicarbazide. In this procedure, this semicarbazide enhanced the reaction of methenamine silver with the glomerular basement membrane and the reaction was completed within a shorter time in comparison with the conventional method. This modification also eliminated any nonspecific reaction with the surface of the glass slide and the solution container and yielded excellent and reproducible results irrespective of the fixation method and material employed. It was also found to stain the renal glomerular basement membrane of rabbits, which is demonstrable only with difficulty by the conventional method.
Animals↗
Enhanced protein phosphorylation, IL-2 receptor expression, and IL-2 production precede cellular proliferation in lymphocytes oxidized by periodic acid.
Increased rates of protein phosphorylation, IL-2 receptor (IL-2R) expression, IL-2 production and DNA synthesis were quantified in rat lymphocytes oxidized by periodic acid (H5IO6). Enhanced phosphorylation of 97, 59 and 37 to 29 kDa proteins in lymphocytes was detected at 18 and 36 hrs post-oxidation, whereas IL-2R expression and IL-2 elaboration were at their maximum by 24 hrs. The number of oxidized cells entering the S-phase of the cell cycle and their thymidine incorporation rates reached their maximum at 72 hrs. These results indicate that H5IO6-induced blastogenesis elicits a progression of responses temporally similar to those of lymphocytes stimulated by lectins.
Animals↗
Periodic acid methenamine silver (pam) staining of the human subcutaneous lymphatic vessel.
The fine structure of the subcutaneous lymphatic vessel taken from the dorsum pedis of the human adult was investigated without and after impregnation by periodic acid methenamine silver (PAM). To check the stainability of another tissue, the renal glomerulus of the rat was also examined in the same way as outlined above. The membrane system of various cells stains positive. No special surface coat of the endothelial cell of the lymphatic vessel was detected. The fine intracellular filament as well as a continuous basal lamina of the endothelial cell were almost PAM-negative, although a part of the basal lamina was occasionally weakly positive. The chemical composition of the basal lamina of the lymphatic vessel is considered to differ from that of the glomerular blood capillary of the kidney, which is PAM-positive. This difference is thought to be due to the smaller polysaccharide content in the former than in the latter.
Adult↗
Pararosaniline or acriflavine-Schiff staining of epoxy embedded tissue after periodic acid oxidation in ethanol: a method suitable for morphometric and fluorometric analysis of glycogen.
A method for preparing tissue sections for automatic image analysis of glycogen is described. Large semithin sections of epoxy embedded tissue fixed in glutaraldehyde-osmium were stained with Schiff reagent and acriflavine (fluorescent staining) after resin removal and periodic acid oxidation in ethanol. We found it essential to avoid tissue rehydration before final staining. The Schiff stain permits an assessment of the cellular volume of glycogen, and the acriflavine allows a fluorometric evaluation of glycogen density.
Acriflavine↗
Periodic acid thiosemicarbazide gelatin methenamine silver (PATSC-GMS) staining for transmission electron microscopy.
A new electron microscopic method is described in which superior staining of basement membranes, collagen fibers and glycogen particles in various tissues was achieved. This method uses a gelatin methenamine silver solution applied to the ultrathin sections on nickel grids after oxidation in a periodic acid. The staining results were excellent and tissue components were evenly stained with no background silver grains. This method proved to be consistent, reproducible and reliable.
Animals↗
Senile plaque-like structures: observation of a probably unknown type of senile plaque by periodic-acid methenamine silver (PAM) electron microscopy.
Numerous diffuse senile plaque-like structures (SPLSs) were found in the cerebral cortex from cases with dementia of the Alzheimer type by means of the methenamine-Bodian method. SPLSs varied in shape and size. They were never recognized in the original Bodian, PAS and Congo red preparations, but were positive with anti-beta-protein immunostaining and periodic-acid methenamine silver (PAM) methods, which are thought to specifically stain amyloid substance. With PAM electron microscopy, we found sparse aggregations of amorphous, often ramified, structures with fine granular silver deposits in SPLS. Routine electron microscopic examination on the same portion where SPLS were confirmed by PAM electron microscopy revealed amorphous, partially fibrous structures. These structures might be amyloid or amyloid-precursor substance. In SPLSs only a few degenerated neurites and astrocytic processes with glycogen granules were seen. We consider SPLSs to be a kind of senile plaque.
Adult↗
[Mechanism of lymphocyte activation by periodic acids. Blastogenesis and induction of immunomodulator populations].
Mild periodate treatment is mitogenic for T lymphocytes. With murine spleen cells periodate oxidation is effective between C8 and C9 on sialyl acid residues. With human blood lymphocytes this oxidation occurs between C7 and C8 of these residues. In vitro immune response is inhibited by periodate treatment. Activation of an immunosuppressive T lymphocyte population is obtained. Similar results are performed with human blood lymphocytes.
Animals↗
Periodic acid--methenamine silver strain for demonstration of cells in Gaucher's disease.
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Gaucher Disease↗