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Urinary and faecal N-methylhistamine concentrations do not serve as markers for mast cell activation or clinical disease activity in dogs with chronic enteropathies.

Anfinsen KP, Berghoff N, Priestnall SL, Suchodolski JS, Steiner JM, Allenspach K - Acta Vet. Scand. (2014)

Bottom Line: Post hoc analysis revealed a statistically significant difference in toluidine blue positive mast cells between two treatment groups (exclusion diet with/without metronidazole versus immunosuppression (IS)), with higher numbers among dogs not requiring IS.The number of duodenal mast cells was higher in dogs that did not need IS, i.e. in dogs responding to an exclusion diet (with/without metronidazole), than in dogs requiring IS.Further studies comparing the role of mast cells in dogs with different forms of CE are needed.

View Article: PubMed Central - PubMed

Affiliation: Department of Veterinary Clinical Sciences, Royal Veterinary College, University of London, Hatfield, AL9 7TA, England. kristin.anfinsen@nmbu.no.

ABSTRACT

Background: This study sought to correlate faecal and urinary N-methylhistamine (NMH) concentrations with resting versus degranulated duodenal mast cell numbers in dogs with chronic enteropathies (CE), and investigate correlations between intestinal mast cell activation and clinical severity of disease as assessed by canine chronic enteropathy clinical activity index (CCECAI), and between urinary and faecal NMH concentrations, mast cell numbers, and histopathological scores. Twenty-eight dogs with CE were included. Duodenal biopsies were stained with haematoxylin and eosin (H&E), toluidine blue, and by immunohistochemical labelling for tryptase. Duodenal biopsies were assigned a histopathological severity score, and duodenal mast cell numbers were counted in five high-power fields after metachromatic and immunohistochemical staining. Faecal and urinary NMH concentrations were measured by gas chromatography-mass spectrometry.

Results: There was no correlation between the CCECAI and faecal or urinary NMH concentrations, mast cell numbers, or histopathological score - or between faecal or urinary NMH concentration and mast cell numbers. Post hoc analysis revealed a statistically significant difference in toluidine blue positive mast cells between two treatment groups (exclusion diet with/without metronidazole versus immunosuppression (IS)), with higher numbers among dogs not requiring IS.

Conclusion: Faecal and urinary NMH concentrations and duodenal mast cell numbers were not useful indicators of severity of disease as assessed by the CCECAI or histological evaluation. The number of duodenal mast cells was higher in dogs that did not need IS, i.e. in dogs responding to an exclusion diet (with/without metronidazole), than in dogs requiring IS. Further studies comparing the role of mast cells in dogs with different forms of CE are needed.

No MeSH data available.


Related in: MedlinePlus

Duodenal section from a dog with chronic enteropathy stained with toluidine blue for metachromatic staining of mast cells (×200). Purple cells represent mast cells. Bar = 100 μm.
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Fig1: Duodenal section from a dog with chronic enteropathy stained with toluidine blue for metachromatic staining of mast cells (×200). Purple cells represent mast cells. Bar = 100 μm.

Mentions: Metachromatic staining with toluidine blue was performed as described elsewhere [12] (Figure 1). Immunohistochemical staining for mast cell tryptase was performed using an automated staining machine (Leica BondMax) and amplification kit (Leica Refine), according to the standard immunohistochemical protocol used at the RVC Diagnostic Laboratory (Figure 2). Briefly, 4 μm sections of duodenal mucosa were pretreated with a pH 9.0 retrieval solution (Dako Epitope Retrieval 2 solution, Dako, Ely) for 10 minutes then a mouse anti-human mast cell tryptase monoclonal antibody (Dako) was used at a 1:800 dilution. Sections were counterstained with haematoxylin.Figure 1


Urinary and faecal N-methylhistamine concentrations do not serve as markers for mast cell activation or clinical disease activity in dogs with chronic enteropathies.

Anfinsen KP, Berghoff N, Priestnall SL, Suchodolski JS, Steiner JM, Allenspach K - Acta Vet. Scand. (2014)

Duodenal section from a dog with chronic enteropathy stained with toluidine blue for metachromatic staining of mast cells (×200). Purple cells represent mast cells. Bar = 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4288550&req=5

Fig1: Duodenal section from a dog with chronic enteropathy stained with toluidine blue for metachromatic staining of mast cells (×200). Purple cells represent mast cells. Bar = 100 μm.
Mentions: Metachromatic staining with toluidine blue was performed as described elsewhere [12] (Figure 1). Immunohistochemical staining for mast cell tryptase was performed using an automated staining machine (Leica BondMax) and amplification kit (Leica Refine), according to the standard immunohistochemical protocol used at the RVC Diagnostic Laboratory (Figure 2). Briefly, 4 μm sections of duodenal mucosa were pretreated with a pH 9.0 retrieval solution (Dako Epitope Retrieval 2 solution, Dako, Ely) for 10 minutes then a mouse anti-human mast cell tryptase monoclonal antibody (Dako) was used at a 1:800 dilution. Sections were counterstained with haematoxylin.Figure 1

Bottom Line: Post hoc analysis revealed a statistically significant difference in toluidine blue positive mast cells between two treatment groups (exclusion diet with/without metronidazole versus immunosuppression (IS)), with higher numbers among dogs not requiring IS.The number of duodenal mast cells was higher in dogs that did not need IS, i.e. in dogs responding to an exclusion diet (with/without metronidazole), than in dogs requiring IS.Further studies comparing the role of mast cells in dogs with different forms of CE are needed.

View Article: PubMed Central - PubMed

Affiliation: Department of Veterinary Clinical Sciences, Royal Veterinary College, University of London, Hatfield, AL9 7TA, England. kristin.anfinsen@nmbu.no.

ABSTRACT

Background: This study sought to correlate faecal and urinary N-methylhistamine (NMH) concentrations with resting versus degranulated duodenal mast cell numbers in dogs with chronic enteropathies (CE), and investigate correlations between intestinal mast cell activation and clinical severity of disease as assessed by canine chronic enteropathy clinical activity index (CCECAI), and between urinary and faecal NMH concentrations, mast cell numbers, and histopathological scores. Twenty-eight dogs with CE were included. Duodenal biopsies were stained with haematoxylin and eosin (H&E), toluidine blue, and by immunohistochemical labelling for tryptase. Duodenal biopsies were assigned a histopathological severity score, and duodenal mast cell numbers were counted in five high-power fields after metachromatic and immunohistochemical staining. Faecal and urinary NMH concentrations were measured by gas chromatography-mass spectrometry.

Results: There was no correlation between the CCECAI and faecal or urinary NMH concentrations, mast cell numbers, or histopathological score - or between faecal or urinary NMH concentration and mast cell numbers. Post hoc analysis revealed a statistically significant difference in toluidine blue positive mast cells between two treatment groups (exclusion diet with/without metronidazole versus immunosuppression (IS)), with higher numbers among dogs not requiring IS.

Conclusion: Faecal and urinary NMH concentrations and duodenal mast cell numbers were not useful indicators of severity of disease as assessed by the CCECAI or histological evaluation. The number of duodenal mast cells was higher in dogs that did not need IS, i.e. in dogs responding to an exclusion diet (with/without metronidazole), than in dogs requiring IS. Further studies comparing the role of mast cells in dogs with different forms of CE are needed.

No MeSH data available.


Related in: MedlinePlus