scholarly journals A Double Histochemical/Immunohistochemical Staining for the Identification of Canine Mast Cells in Light Microscopy

2021 ◽  
Vol 8 (10) ◽  
pp. 229
Author(s):  
Francesca Gobbo ◽  
Giuseppe Sarli ◽  
Margherita De De Silva ◽  
Giorgia Galiazzo ◽  
Roberto Chiocchetti ◽  
...  
Keyword(s):  
Mast Cells ◽  
Cannabinoid Receptor ◽  
Immunohistochemical Analysis ◽  
Inflammatory Cells ◽  
Staining Technique ◽  
Cannabinoid Receptor 2 ◽  
Mast Cell Tumors ◽  
Immunohistochemical Markers ◽  
Poorly Differentiated ◽  
Cutaneous Mastocytosis

Immunohistochemistry (IHC) is a widely used technique in diagnostic pathology, but the simultaneous analysis of more than one antibody at a time with different chromogens is rather complex, time-consuming, and quite expensive. In order to facilitate the identification of mast cells (MCs) during immunohistochemical analysis of membrane and/or nuclear markers, we propose a new staining method that includes the association of IHC and toluidine blue as a counterstain. To achieve this goal, we tested c-kit, Ki67, and cannabinoid receptor 2 on several cases of cutaneous canine mast cell tumors (MCTs), cutaneous mastocytosis, and atopic dermatitis. The results obtained show how this double staining technique, although limited to non-cytoplasmic markers and of little use in poorly differentiated MCTs in which MC metachromasia is hard to see, can be used during the evaluation of nuclear and/or membranous immunohistochemical markers in all canine cutaneous disorders, especially if characterized by the presence of a low number of MCs. It can help to evaluate those MCTs in which neoplastic MCs must be clearly distinguished from inflammatory cells that can infiltrate the tumor itself, in facilitating the calculation of the Ki67 index. Moreover, it can be used to study the expression of new markers in both animal and human tissues containing MCs and in MC disorders.

scholarly journals Chymase and tryptase in dog mastocytoma cells: asynchronous expression as revealed by enzyme cytochemical staining.

1988 ◽  
Vol 36 (8) ◽  
pp. 1053-1060 ◽  
Author(s):  
G H Caughey ◽  
N F Viro ◽  
L D Calonico ◽  
D M McDonald ◽  
S C Lazarus ◽  
...  
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Staining Technique ◽  
Staining Intensity ◽  
Cell Development ◽  
Cytochemical Staining ◽  
Mast Cell Tumors ◽  
General Utility ◽  
Developmental Relationship ◽  
Mastocytoma Cells

Mast cell populations can be distinguished by differences in the content and substrate specificity of their two major cytoplasmic granule proteases, the chymases and the tryptases. To explore the origins of differences in the types of proteases present in mast cells, we used a double cytochemical staining technique to reveal both chymase and tryptase in cells from four lines of dog mast cell tumors containing both enzymes. We expected that if chymase and tryptase were expressed together during cell development the relative staining intensity of chymase compared to tryptase would be constant among different cells of each tumor. Instead, we found substantial variation in the relative intensity of chymase and tryptase staining among cells of a given mastocytoma line, each of which contained cells presumed to be monoclonal in origin but heterogeneous with respect to cell development. The overall staining intensity for chymase or tryptase correlated with the amount of protease activity in extracts of tumor homogenates. Staining specificity was established by use of selective inhibitors and competitive substrates and was tested on various types of dog cells obtained by bronchoalveolar lavage. The results suggest that active chymase and tryptase may be expressed differently during mast cell differentiation and support the possibility of a close developmental relationship between mast cells differing in protease phenotype. Moreover, the success of the staining procedures applied to mastocytoma cells suggests that they may be of general utility in phenotyping of mast cells according to the protease activities present in their granules.

scholarly journals Cannabinoid Receptor 2 Alters Social Memory and Microglial Activity in an Age-Dependent Manner

Molecules ◽  
2021 ◽  
Vol 26 (19) ◽  
pp. 5984
Author(s):  
Joanna Agnieszka Komorowska-Müller ◽  
Tanushka Rana ◽  
Bolanle Fatimat Olabiyi ◽  
Andreas Zimmer ◽  
Anne-Caroline Schmöle
Keyword(s):  
Cannabinoid Receptor ◽  
Social Memory ◽  
Brain Aging ◽  
Endocannabinoid System ◽  
Immunohistochemical Analysis ◽  
Cellular Level ◽  
Spatial Learning And Memory ◽  
Dependent Manner ◽  
Cannabinoid Receptor 2 ◽  
Age Related

Physiological brain aging is characterized by gradual, substantial changes in cognitive ability, accompanied by chronic activation of the neural immune system. This form of inflammation, termed inflammaging, in the central nervous system is primarily enacted through microglia, the resident immune cells. The endocannabinoid system, and particularly the cannabinoid receptor 2 (CB2R), is a major regulator of the activity of microglia and is upregulated under inflammatory conditions. Here, we elucidated the role of the CB2R in physiological brain aging. We used CB2R−/− mice of progressive ages in a behavioral test battery to assess social and spatial learning and memory. This was followed by detailed immunohistochemical analysis of microglial activity and morphology, and of the expression of pro-inflammatory cytokines in the hippocampus. CB2R deletion decreased social memory in young mice, but did not affect spatial memory. In fact, old CB2R−/− mice had a slightly improved social memory, whereas in WT mice we detected an age-related cognitive decline. On a cellular level, CB2R deletion increased lipofuscin accumulation in microglia, but not in neurons. CB2R−/− microglia showed an increase of activity markers Iba1 and CD68, and minor upregulation in tnfa and il6 expression and downregulation of ccl2 with age. This was accompanied by a change in morphology as CB2R−/− microglia had smaller somas and lower polarity, with increased branching, cell volume, and tree length. We present that CB2Rs are involved in cognition and age-induced microglial activity, but may also be important for microglial activation itself.

Equine Cutaneous Mastocytosis

1970 ◽  
Vol 7 (1) ◽  
pp. 43-55 ◽  
Author(s):  
K. Altera ◽  
L. Clark
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Fibrinoid Necrosis ◽  
Cutaneous Lesions ◽  
Collagen Diseases ◽  
Focal Necrosis ◽  
Mast Cell Tumors ◽  
Cutaneous Mast Cell ◽  
Cutaneous Mastocytosis ◽  
Necrotic Debris

A description of equine cutaneous mastocytosis is given. The lesion was characterized by focal aggregates of mast cells, and by eosinophils, fibrinoid necrosis of collagen, and focal necrosis with dystrophic mineralization of necrotic debris. This is an uncommon, previously undescribed lesion in horses. The authors encountered 14 cases in a 16-year period. The etiology and nature of the lesion are undetermined. Morphologic similarities to canine cutaneous mast-cell tumors, cutaneous lesions of collagen diseases, and calcinosis circumscripta are noted and the possibility of relationships to these lesions is discussed.

Increased levels of lysosomal cysteinyl cathepsins in human varicose veins: A histology study

2014 ◽  
Vol 111 (02) ◽  
pp. 333-344 ◽  
Author(s):  
Na Xu ◽  
Yuan-Yuan Zhang ◽  
Yan Lin ◽  
Bin Bao ◽  
Lei Zheng ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
T Cells ◽  
Mast Cells ◽  
Cystatin C ◽  
Varicose Veins ◽  
Immunohistochemical Analysis ◽  
Inflammatory Cells ◽  
Cysteine Proteases ◽  
Chronic Venous Disease ◽  
Venous Disease

SummaryVaricose veins are a major chronic venous disease characterised by extensive remodelling of the extracellular matrix architecture in the vascular wall. Although matrix metalloproteinases have been implicated in these pathologic events, little is known about the functional relevance of other protease family members. Here, we studied the distribution of lysosomal cysteine proteases, cathepsins B, L, K, and S, and their endogenous inhibitor, cystatin C, in long saphenous vein specimens from nine normal donors and 18 patients with varicose veins (VVs). Immunohistochemical analysis demonstrated increased levels of cathepsins L, K, B, and S and reduced levels of cystatin C in VVs. This imbalance between cysteinyl cathepsins and cystatin C may favour VV remodelling. To investigate the inflammatory mechanism of their expression, we examined a detailed inflammatory cell profile in VVs, including macrophages, T lymphocytes, and mast cells. Increased numbers of CD3-positive T cells and tryptase-positive mast cells were found in VVs, and enhanced levels of cysteinyl cathepsins were detected from lesion CD3-positive T cells, chymase-positive mast cells, endothelial cells, and smooth-muscle cells. Elevated cathepsins, and their co-localisation to infiltrated inflammatory cells and to vascular cells, suggest that these proteases participate in extracellular matrix degradation in response to inflammation during VV pathogenesis.

scholarly journals Optical Control of Cannabinoid Receptor 2-Mediated Ca2+ Release Enabled by Synthesis of Photoswitchable Probes

2021 ◽  
Author(s):  
Roman C. Sarott ◽  
Alexander E. G. Viray ◽  
Patrick Pfaff ◽  
Anastasiia Sadybekov ◽  
Gabriela Rajic ◽  
...  
Keyword(s):  
Cannabinoid Receptor ◽  
Optical Control ◽  
Cannabinoid Receptor 2

scholarly journals The Role of Immunohistochemical Markers for the Diagnosis and Prognosis of Adrenocortical Neoplasms

2021 ◽  
Vol 11 (3) ◽  
pp. 208
Author(s):  
Anna Angelousi ◽  
Georgios Kyriakopoulos ◽  
Fani Athanasouli ◽  
Anastasia Dimitriadi ◽  
Eva Kassi ◽  
...  
Keyword(s):  
Transcriptome Analysis ◽  
Cox Regression ◽  
P53 Protein ◽  
Immunohistochemical Analysis ◽  
Ki 67 ◽  
Diagnosis And Prognosis ◽  
Immunohistochemical Markers ◽  
Small Series ◽  
Morphological Criteria ◽  
Survival And Development

Adrenal cortical carcinoma (ACC) is a rare cancer with poor prognosis that needs to be distinguished from adrenocortical adenomas (ACAs). Although, the recently developed transcriptome analysis seems to be a reliable tool for the differential diagnosis of adrenocortical neoplasms, it is not widely available in clinical practice. We aim to evaluate histological and immunohistochemical markers for the distinction of ACCs from ACAs along with assessing their prognostic role. Clinical data were retrospectively analyzed from 37 patients; 24 archived, formalin-fixed, and paraffin-embedded ACC samples underwent histochemical analysis of reticulin and immunohistochemical analysis of p27, p53, Ki-67 markers and were compared with 13 ACA samples. Weiss and Helsinki scores were also considered. Kaplan−Meier and univariate Cox regression methods were implemented to identify prognostic effects. Altered reticulin pattern, Ki-67% labelling index and overexpression of p53 protein were found to be useful histopathological markers for distinguishing ACAs from ACCs. Among the studied markers, only pathological p53 nuclear protein expression was found to reach statistically significant association with poor survival and development of metastases, although in a small series of patients. In conclusion, altered reticulin pattern and p53/Ki-67 expression are useful markers for distinguishing ACCs from ACAs. Immunohistopathology alone cannot discriminate ACCs with different prognosis and it should be combined with morphological criteria and transcriptome analysis.

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