scholarly journals Glycol methacrylate embedding for the histochemical study of the gastrointestinal tract of dogs naturally infected with Leishmania infantum

2015 ◽  
Vol 59 (4) ◽  
Author(s):  
A.J.W. Pinto ◽  
I.F.G. De Amorim ◽  
L.J. Pinheiro ◽  
I.M.V.M. Madeira ◽  
C.C. Souza ◽  
...  
Keyword(s):  
Mast Cell ◽  
Gastrointestinal Tract ◽  
Toluidine Blue ◽  
Cell Morphology ◽  
Histological Study ◽  
Periodic Acid ◽  
Clinical Status ◽  
Parasite Load ◽  
Periodic Acid Schiff ◽  
Glycol Methacrylate

<p>In canine visceral leishmaniasis a diffuse chronic inflammatory exudate and an intense parasite load throughout the gastrointestinal tract has been previously reported. However, these studies did not allow a properly description of canine cellular morphology details. The aim of our study was to better characterize these cells in carrying out a qualitative and quantitative histological study in the gastrointestinal tract of dogs naturally infected with <em>Leishmania</em> <em>infantum </em>by examining gut tissues embedded in glycol methacrylate. Twelve infected adult dogs were classified in asymptomatic and symptomatic. Five uninfected dogs were used as controls. After necropsy, three samples of each gut segment, including esophagus, stomach, duodenum, jejunum, ileum, cecum, colon, and rectum were collected and fixed in Carnoy’s solution for glycol methacrylate protocols. Sections were stained with hematoxylin-eosin, toluidine blue borate, and periodic acid–Schiff stain. <em>Leishmania</em> amastigotes were detected by immunohistochemistry employed in both glycol methacrylate and paraffin embedded tissues. The quantitative histological analysis showed higher numbers of plasma cells, lymphocytes and macrophages in lamina propria of all segments of GIT of infected dogs than controls. The parasite load was more intense and cecum and colon, independently of the clinical status of these dogs. Importantly, glycol methacrylate embedded tissue stained with toluidine blue borate clearly revealed mast cell morphology, even after mast cell degranulation. Infected dogs showed lower numbers of mast cells in all gut segments than did controls.  Despite the glycol methacrylate (GMA) protocol requires more attention and care than the conventional paraffin processing, this embedding procedure proved to be especially suitable for the present histological study, where it allowed to preserve and observe cell morphology in fine detail.</p>

scholarly journals Canine Mast Cell Tumors: A Comparison of Staining Techniques

1994 ◽  
Vol 6 (4) ◽  
pp. 458-465 ◽  
Author(s):  
Jose Pedro Cannas Simoes ◽  
Polly Schoning
Keyword(s):  
Mast Cell ◽  
Toluidine Blue ◽  
Periodic Acid ◽  
Relative Effectiveness ◽  
Periodic Acid Schiff ◽  
Additional Information ◽  
Histochemical Methods ◽  
Mast Cell Tumors ◽  
Staining Methods ◽  
Safranin O

Twelve histochemical methods; affinity staining with avidin peroxidase, wheat germ agglutinin, and concavalin-A agglutinin; and an immunohistochemical stain with Kpl (CD68) antibody were compared for their relative effectiveness in staining canine mast cell tumors. Stains were compared in 28 mast cell tumors and 19 histiocytomas. The effectiveness of the histochemical methods and the lectins decreased as the mast cells became less differentiated. None of the staining methods were positive on histiocytomas. Periodic acid-Schiff (PAS) gave positive results in a few cases of mast cell tumors where other histochemical stains were negative. Although avidin peroxidase and Kpl antibody stained more mast cell tumors than any other method, they did not differ significantly from Luna's method, toluidine blue pH 0.5, toluidine blue pH 4.5, alcian blue pH 2.5, safranin O, Unna's method, and Giemsa. No stain was ideal for the diagnosis of canine mast cell tumors; however, this study suggests that the use of avidin peroxidase, Kpl antibody, and PAS may give additional information for individual poorly differentiated tumors without substantial increase in time or cost.

scholarly journals THE HISTOCHEMICAL INTERPRETATION OF THE COMPLEX RESULTS OF METHYLATION UPON GASTROINTESTINAL TRACT MUCINS, WITH SPECIAL REFERENCE TO THE PERIODIC ACID-SCHIFF REACTIVITY

1974 ◽  
Vol 22 (10) ◽  
pp. 986-991 ◽  
Author(s):  
P. E. REID ◽  
C. F. A. CULLING ◽  
W. L. DUNN
Keyword(s):  
Sialic Acid ◽  
Gastrointestinal Tract ◽  
Special Reference ◽  
Periodic Acid ◽  
Separate Study ◽  
Sulfate Ester ◽  
Periodic Acid Schiff ◽  
Smith Degradation ◽  
Vicinal Diol ◽  
Schiff Reaction

The histochemical use of methylation has complex results; particularly in respect of the periodic acid-Schiff reaction, these are analyzed and discussed. Methods are described which allow the separate study of the following effects: (a) the removal of the KOH/periodic acid-Schiff effect; (b) removal of sialic acid from a potential vicinal diol; and (c) the removal of O-sulfate ester from a potential vicinal diol. The use of the Smith degradation technique, in addition to the above, also allows inferences to be drawn in respect of the structure of the mucins (glycoproteins) being investigated.

A physiological, biochemical and histological study of goose tracheal mucin and its secretion

1977 ◽  
Vol 279 (969) ◽  
pp. 513-540 ◽  
Keyword(s):  
Sialic Acid ◽  
Nerve Stimulation ◽  
Histological Study ◽  
Periodic Acid ◽  
Gel Filtration ◽  
Goblet Cells ◽  
Mucin Secretion ◽  
Electron Microscopic ◽  
Periodic Acid Schiff ◽  
Tracheal Mucus

Tracheal mucin secretion has been measured from a segment of trachea, isolated in situ , in anaesthetized geese by a method that involves radioactive labelling of tracheal mucus glycoproteins (Gallagher et al. 1975). Goose tracheal mucus comes entirely from goblet cells, since the goose trachea does not contain submucosal mucous or serous glands, and this method has been used to investigate the nervous and pharmacological control of the mucin secretion from these epithelial goblet cells. The mucins secreted have been collected, fractionated, and chemically analysed. Intracellular mucin has been examined histochemically, and the results of electron microscopic observations of epithelial cells and nerves are presented. Acetylcholine increased tracheal mucin secretion, and this effect was completely blocked by atropine. Neither α- nor β-stimulant sympathomimetic amines affected tracheal mucin secretion. Stimulation of the peripheral cut ends of the descending oesophageal nerves increased tracheal mucin secretion and the majority of this response, approximately three-quarters, appeared to be cholinergic since this proportion was blocked by atropine. The mediator for the atropine-resistant part of the response is not known, but it appears not to be a β-adrenoreceptor stimulant since the response to nerve stimulation was unaffected by propranolol given at 34 μm intrasegmentally. Other possibilities are discussed. Atropine itself decreased the resting level of tracheal mucin secretion. The local anaesthetic, lignocaine, increased tracheal mucin secretion, while at the same time blocking the responses to acetylcholine and descending oesophageal nerve stimulation. The implications of this are discussed. The electrophoretic, gel filtration and ion-exchange properties of goose tracheal mucins showed that they represented high molecular mass, negatively charged glycoproteins which could be labelled biosynthetically with [ 35 S]sulphate, [ 3 H]- and [ 14 C]glucose. These mucins could be stained with Alcian blue or periodic acid Schiff reagent. The carbohydrate composition was unusual for an epithelial glycoprotein in that fucose was absent and mannose was present in small quantities. The monosaccharides present in larger quantity were galactose, N -acetylglucosamine, N -acetylgalactosamine and sialic acid. Histochemical analysis of tissue sections of gosling tracheas demonstrated that nearly all of the glycoprotein in epithelial goblet cells contained both sialic acid and sulphate residues. Sialated mucin was present also, but to a lesser extent, and many cells contained a mixture of sialated and sulphated mucins. The adult goose trachea had a high proportion of sialated glycoprotein. Electron microscopy showed a range of epithelial cell types and intra-epithelial nerves also. Many of the nerves had neurosecretory vesicles suggestive of motor function and some were near to goblet cells.

scholarly journals Histochemical and Autoradiographic Studies on the Effects of Aging on the Mucopolysaccharides of the Periosteum

1959 ◽  
Vol 6 (2) ◽  
pp. 171-178 ◽  
Author(s):  
Edgar A. Tonna ◽  
Eugene P. Cronkite
Keyword(s):  
Chondroitin Sulfate ◽  
Toluidine Blue ◽  
Bone Growth ◽  
Periodic Acid ◽  
Neutral Type ◽  
Autoradiographic Study ◽  
Colloidal Iron ◽  
Periodic Acid Schiff ◽  
Schiff Reaction ◽  
Effects Of Aging

An autoradiographic study was made using S35-sulfate for the localization, distribution, and variation in the mucopolysaccharide content of the femoral periosteum of rats from birth to old age. The mucopolysaccharides were also studied histochemically, using toluidine blue O, Rinehart and Abu'l-Haj's colloidal iron method, and the periodic acid-Schiff reaction, before and after hyaluronidase treatment. Autoradiograms revealed the uptake of S35 particularly in the vicinity of the preosseous zone and adjacent osteoblasts. This labelling was highest at the period of rapid bone growth. With increasing age, the S35 uptake became progressively less. The preosseous zone showed γ-metachromatic staining at all ages after treatment with toluidine blue. Active osteoblasts were mostly orthochromatic, however, ß-metachromasia was exhibited at a later age. Abundant amounts of intra- and extracellular mucopolysaccharides of both the acid and neutral type were demonstrated in the periosteum. S35 uptake and γ-metachromasia show the presence of sulfated mucopolysaccharides, of which chondroitin sulfate predominates in the preosseous zone. Since S35 uptake is high in active osteoblasts, the inability to demonstrate metachromasia in osteoblasts may indicate either that chondroitin sulfate is liberated as fast as it is being produced, or that it may be present within the cells in a precursor form not detectable by histochemical methods.

scholarly journals METACHROMASIA INHIBITING COMPONENTS IN AMYLOID

1958 ◽  
Vol 6 (3) ◽  
pp. 181-184 ◽  
Author(s):  
BØRGE LARSEN
Keyword(s):  
Chondroitin Sulfate ◽  
Toluidine Blue ◽  
Serum Proteins ◽  
Periodic Acid ◽  
Methyl Violet ◽  
Inhibitory Effect ◽  
Blue Staining ◽  
Periodic Acid Schiff ◽  
Toluidine Blue Staining ◽  
Tissue Sections

I): The degree of inhibitory effect of serum proteins and a periodic acid Schiff-positive amyloid fraction on metachromasia resulting from a metachromatic amyloid fraction was studied and compared to the metachromatic potency of chondroitin sulfate exposed to the same type of inhibition. The metachromatic properties of chondroitin sulfate were close to those of the metachromatic amyloid fraction. II): In comparing the metachromasia obtained with toluidine blue and methyl violet, it was noted that on incubation with the periodic acid Schiff-positive amyloid fraction, amyloid metachromosia after methyl violet staining was inhibited less than after toluidine blue staining. III): It is emphasized that a competition between dye and proteins occurs in metachromatic staining reactions. IV): Inhibition of amyloid metachromasia in histological tissue sections could not be demonstrated with the techniques used.

Mast cell infiltration associated with tubulointerstitial fibrosis in chronic Aristolochic Acid Nephropathy

2005 ◽  
Vol 24 (2) ◽  
pp. 41-47 ◽  
Author(s):  
Yichao Wu ◽  
Zhihong Liu ◽  
Weixin Hu ◽  
Leishi Li
Keyword(s):  
Mast Cell ◽  
Aristolochic Acid ◽  
Interstitial Fibrosis ◽  
Smooth Muscle Actin ◽  
Periodic Acid ◽  
Basement Membranes ◽  
Cell Infiltration ◽  
Alpha Smooth Muscle Actin ◽  
Periodic Acid Schiff ◽  
Aristolochic Acid Nephropathy

Aristolochic Acid Nephropathy (AAN) is regarded as a kind of toxic nephropathy caused by the formation of DNA- aristolochic acid adducts in renal parenchymal cells. However, the underlying mechanisms driving the progression of renal interstitial fibrosis in AAN still remains unclear. This study aims to elucidate the role of some immunological factors, especially mast cells (MCs), in the pathogenesis of AAN. Sixteen patients with AAN were enrolled in this study, including five acute and 11 chronic AAN. Monoclonal antibodies against human tryptase, alpha smooth muscle actin (α-SMA), and CD68 were applied on serial sections, which were further counterstained with Periodic Acid-Schiff. It was found that massive tryptase-positive MCs were observed in the fibrotic areas in chronic AAN, especially around thickened tubular basement membranes where myofibroblasts accumulated too. In contrast, MCs infiltrated to a less extent in acute AAN, and were barely found in normal control kidneys. In chronic AAN, the number of MCs in the tubulointerstitium was positively correlated with the degree of renal fibrosis ( r=0.64, P<0.05), but not with serum creatinine levels. Meanwhile, the recruitment of MCs into the renal interstitium is accompanied with local proliferation of myofibroblasts. Macrophages were not abundant, neither in acute nor in chronic AAN. Our findings show for the first time that mast cell infiltration seems to be associated with the progression of fibrosis in the renal tubulointerstitium in chronic AAN.

scholarly journals The presence of mast cell precursors in rat peripheral blood

Blood ◽  
1981 ◽  
Vol 58 (3) ◽  
pp. 544-551 ◽  
Author(s):  
D Zucker-Franklin ◽  
G Grusky ◽  
N Hirayama ◽  
E Schnipper
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Alcian Blue ◽  
Peripheral Blood ◽  
Periodic Acid ◽  
Phenotypic Expression ◽  
Soft Agar ◽  
Sulfated Polysaccharides ◽  
Periodic Acid Schiff ◽  
Agar Culture

Abstract Soft agar culture of mononuclear cell fractions prepared from rat peripheral blood yielded numerous colonies consisting of mast cells. The mast cell nature of the cells was established by ultrastructural and histochemical analyses as well as by the demonstration the the colonies contained histamine and that the cells possessed receptors for the Fc component of IgE. Stringent criteria for the distinction of mast cells from monocytes/macrophages that could have metachromatic inclusions were applied. The alcian-blue-safranin technique delineated the maturation of mast cell granules by showing the loss of alcian-blue and increase in safranin-positive organelles presumed to reflect the increase in N-sulfated polysaccharides representing heparin. The mast cells exhibited low or absent reactions for peroxidase, alpha-naphthyl butyrate, periodic acid Schiff, and Sudan black reacting lipid, whereas macrophages stained in parallel were positive for these substances. Since it is known that extracellular conditions may cause variations in phenotypic expression, the observations have led to the hypothesis that mast cells and macrophages may have a common precursor.

Trophoblast ultrastructure in murine interspecies pregnancy

1988 ◽  
Vol 46 ◽  
pp. 286-287
Author(s):  
M. Crepeau ◽  
S. Yamashiro ◽  
T. Bast ◽  
B.A. Croy
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Mus Musculus ◽  
Histological Study ◽  
Periodic Acid ◽  
Periodic Acid Schiff ◽  
Pregnancy Failure ◽  
Mus Caroli ◽  
Implantation Sites ◽  
Time Point

Mus caroli blastocysts transferred to the uterus of pseudopregnant Mus musculus implant but do not survive past midgestation. Blastocyst microsurgery has been used to show that trophoblast is the tissue responsible for pregnancy failure but the nature of the failure remains undefined. Therefore a histological study, was undertaken to compare development of M. caroli embryos in M. musculus and M. caroli uteri at a time point just prior to development of grossly observable lesions.Seven transferred M. caroli embryos and six transferred M. musculus embryos were recovered from three M. musculus recipients on day 8.5 of gestation. Four M. caroli embryos recovered from a M. caroli uterus on day 7.5 of gestation were used as stage-matched controls. For light microscopy, 2 μm thick, glycol methacrylate embedded (3), sections of implantation sites were stained with H & E or periodic acid-Schiff (PAS). For electron microscopy, small pieces of the tissues were fixed and processed routinely.

scholarly journals Histology and Mucous Histochemistry of the Integument and Body Wall of a Marine Polychaete Worm,Ophryotrochan. sp. (Annelida: Dorvilleidae) Associated with Steelhead Trout Cage Sites on the South Coast of Newfoundland

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
H. M. Murray ◽  
D. Gallardi ◽  
Y. S. Gidge ◽  
G. L. Sheppard
Keyword(s):  
Alcian Blue ◽  
Body Wall ◽  
Histological Study ◽  
Periodic Acid ◽  
The Body ◽  
South Coast ◽  
Steelhead Trout ◽  
The South ◽  
Periodic Acid Schiff ◽  
Marine Polychaete

Histology and mucous histochemistry of the integument and body wall of a marine polychaete worm,Ophryotrochan. sp. (Annelida: Dorvilleidae) associated with Steelhead trout cage sites on the south coast of Newfoundland. A new species of polychaete (Ophryotrochan. sp. (Annelida: Dorvilleidae)) was identified from sediment below Steelhead trout cages on the south coast of Newfoundland, Canada. The organisms were observed to produce a network of mucus in which groups of individuals would reside. Questions regarding the nature and cellular source of the mucus were addressed in this study. Samples of worms were taken from below cages and transported to the laboratory where individuals were fixed for histological study of the cuticle and associated mucus histochemistry. The body wall was organized into segments with an outer cuticle that stained strongly for acid mucopolysaccharides. The epidermis was thin and supported by loose fibrous connective tissue layers. Channels separating individual segments were lined with cells staining positive for Alcian blue. Mucoid cellular secretions appeared thick and viscous, strongly staining with Alcian blue and Periodic Acid Schiff Reagent. It was noted that lateral channels were connected via a second channel running through the anterior/posterior axis. The role of mucus secretion is discussed.

scholarly journals EXTRACTION RESISTANCE OF SUDAN STAINED MAST CELLS AFTER PREVIOUS ACID TREATMENT

1964 ◽  
Vol 12 (7) ◽  
pp. 530-532 ◽  
Author(s):  
JOHN R. FEAGLER ◽  
J. F. A. MCMANUS
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Chemical Property ◽  
Cell Nucleus ◽  
Acid Treatment ◽  
Periodic Acid ◽  
Hydrogen Ion ◽  
Periodic Acid Schiff ◽  
Sudan Black B ◽  
Sudan Dye

1. Sudan black B alone stains the mast cells very faintly. The granules do not appear very distinct or numerous. 2. The periodic acid Schiff (PAS) technique stains the mast cells; however, the results are variable. When stained, the mast cell granules appear slightly darker than the cytoplasm. 3. When PAS and Sudan black are used together, the mast cell granules gain an avidity for the dye and are stained black. 4. Brief acid treatment prior to immersion in Sudan black results in granules gaining an avidity for the Sudan dye that is extraction resistant. The reaction appears dependent upon modifying the mast cell granules by acid treatment. It appears as though the hydrogen ion changes some physical or chemical property of the granules so that they gain an avidity for the dye. 5. Acetylated Sudan black shows an affinity for the mast cell granules greater than Sudan black alone. This is not increased by previous acid treatment. Acetylated Sudan black also stains the mast cell nucleus.

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