scholarly journals Quantitation of mast cell heparin by flow cytofluorometry.

1976 ◽  
Vol 24 (12) ◽  
pp. 1231-1238 ◽  
Author(s):  
L Enerbäck ◽  
G Berlin ◽  
I Svensson ◽  
I Rundquist

Mast cells can be automatically identified in a mixed cell population by flow cytofluorometry after Berberine sulphate staining. Volume specific counts of the total number of cells and number of mast cells, as well as frequency distributions of fluorescence intensities of mast cells, based on a large number of cells, can be rapidly obtained. Results obtained by microscope fluorometry of cells identified by phase contrast microscopy showviously published results it may be inferred that the fluorescence intensity of individual mast cells is proportional to mast cell heparin content. The automated cell counts correlated very well with manual hemocytometer counts. Both cell counts and the determination of mean mast cell fluorescence showed excellent reproducibility.

1978 ◽  
Vol 26 (1) ◽  
pp. 14-21 ◽  
Author(s):  
G Berlin ◽  
L Enerbäck

A cytofluorometric method, based on berberine staining of mast cell heparin, was used for flow cytofluorometric counting and heparin quantitation of mast cells in crude peritoneal suspensions of growing rats. The automatic flow cytofluorometric counting of mast cells correlated well with hemocytometer cell counts. The mean mast cell heparin content obtained by flow cytofluorometry showed good agreement with such obtained by cytofluorometry of microscopically identified mast cells. The number of peritoneal mast cells and the mean mast cell heparin content was found to increase as the animals grew older. The results of the microscope fluorometric measurements suggested that the heparin content was normally distributed within mast cell populations of both young and old rats. However, the heparin distributions obtained by flow cytofluorometry were often positively skewed but did not fulfill the condition of the log-normal distribution.


1996 ◽  
Vol 80 (4) ◽  
pp. 1322-1330 ◽  
Author(s):  
M. Longphre ◽  
L. Y. Zhang ◽  
J. R. Harkema ◽  
S. R. Kleeberger

Ozone (O3) exposure produces inflammation in the airways of humans and animal models. However, the mechanism by which O3 affects these changes is uncertain. Mast cells are strategically located below the epithelium of the airways and are capable of releasing a number of proinflammatory mediators. We tested the hypothesis that mast cells contribute to inflammation, epithelial sloughing, and epithelial proliferation in the nasal and terminal bronchiolar murine airways after O3 exposure. Mast cell-sufficient (+/+), mast cell-deficient (W/Wv), and mast cell-repleted [bone marrow-transplanted (BMT) W/Wv] mice were exposed to 2 ppm O3 or filtered air for 3 h. Nasal and bronchoalveolar lavage fluids were collected 6 and 24 h after exposure. Differential cell counts and protein content of the lavage fluids were used as indicators of inflammation and permeability changes in the airways. O3-induced epithelial injury was assessed by light microscopy, and O3-induced DNA synthesis in airway epithelium was estimated by using a 5-bromo-2′-deoxyuridine-labeling index in the nasal and terminal bronchiolar epithelia. Relative to air control mice, O3 caused significant increases in inflammation, epithelial injury, and epithelial DNA synthesis in +/+ mice. There was no significant effect of O3 exposure on any measured parameter in the W/Wv mice. To further assess the role of mast cells in O3-induced epithelial damage, mast cells were restored in W/Wv mice by BMT from +/+ congeners. Relative to sham-transplanted W/Wv mice, O3 caused significant increases in epithelial damage and DNA synthesis as well as inflammatory indicators in BMT W/Wv mice. These observations are consistent with the hypothesis that mast cells significantly modulate the inflammatory and proliferative responses of the murine airways to O3.


1970 ◽  
Vol 49 (3) ◽  
pp. 480-486
Author(s):  
F.M. Sorenson ◽  
J.S. Bennett ◽  
D. Fujita ◽  
F.R. Poindexter ◽  
W.B. Hall

Simple counts of mast cells per unit of human gingiva are often difficult to interpret because of the large numbers and varying sizes and shapes of the counted structures. The relatively simple photoelectric scanning method described herein eliminates tedious counting procedures while providing a measure of the relative quantity of stainable mast cell granules within the area scanned. Thus, the method may provide a better estimate of the total biologic activity than would simple mast cell counts.


2020 ◽  
Vol 12 (02) ◽  
pp. 141-146
Author(s):  
Bhavya P. Mohan ◽  
K.P. Aravindan

Abstract Background and Objective Serotonin levels are increased in acute appendicitis. We investigated the possible source of this increase. The aim of this study was to compare the distribution and density of epithelial and nonepithelial enterochromaffin (EC) cells as well as numbers of degranulated and nondegranulated mast cells in different layers of normal appendices and acute appendicitis. Methods Sections from 15 cases of acute appendicitis and 10 cases where the appendix was morphologically normal were stained with Hematoxylin & Eosin, Toluidine blue, and immunohistochemically for chromogranin and CD-117. EC cells stained by chromogranin were counted per crypt and extraepithelial EC cells counted and expressed as cells per unit area (mm2). Mast cells stained by Toluidine blue and CD-117 were counted in lamina propria, submucosa, and muscle layers. The difference between Toluidine blue and CD117 stained mast cells was taken to be an estimate of degranulated cells. The cell counts were expressed per unit area (mm2) as well as per cross-sectional area of the appendix. Results There was no statistically significant difference in epithelial and extraepithelial EC cells between acute appendicitis and normal appendix. Estimated mast cell degranulation as indicated by mast cell counts per cross-sectional area is greatly increased in acute appendicitis when compared with normal. Conclusion Degranulated mast cells rather than EC cells may be the main source of raised serotonin in acute appendicitis.


1966 ◽  
Vol 33 (1) ◽  
pp. 51-64 ◽  
Author(s):  
L. W. Phipps ◽  
F. H. S. Newbould

SummaryA technique for determining the concentration of leucocytes in cow's milk is described which is more rapid and accurate as well as much less tedious than conventional microscope methods. The leucocytes are isolated from the fat globules of comparable size by a novel centrifuging procedure. Sizing of the leucocytes with a Coulter electronic counter shows that their volumes range from about 45 to 1770 µm3but a partially overlapping distribution of other particulate matter prevents the accurate determination of the total number of cells present. The numbers counted in truncated leucocyte distributions, however, correlate linearly with total cell counts determined with an improved microscope method up to 6 × 106cells/ml of milk.


2020 ◽  
Vol 9 ◽  
pp. 1962
Author(s):  
Soheila Nasiri ◽  
Alireza Salehi ◽  
Azadeh Rakhshan

Background: Alopecia areata (AA) and androgenic alopecia (AGA) are the most common types of alopecias. Recently, the role of mast cells in inflammatory diseases has become the focus of many studies. However, few studies have been conducted on their role in AA and AGA. Therefore, our study aimed to quantitatively evaluate the presence of mast cells in the AA and AGA specimens.Materials and Methods: Three groups of AA, AGA, and healthy control were studied (each group with 20 subjects). Patients were randomly selected from those referred to the dermatology clinics of Shahid Beheshti University. Specimens were obtained from the scalp, and perifollicular and perivascular areas were investigated. Results: Significantly higher perifollicular and perivascular mast cell counts were seen in both AGA and AA groups compared to healthy control (P<0.001 for both). Moreover, AA patients had more frequent perivascular mast cells than the AGA group (P=0.042). Among patients aged <40 years, perifollicular and perivascular mast cell counts were not significantly different among the three groups; however, subjects over 40 years of age in both groups had significantly more perifollicular and perivascular mast cells than healthy participants. There was a significant positive correlation between disease severity and mast cell counts in both perifollicular and perivascular areas in AA patients (P=0.001 for both). Conclusion: There was a significantly increased infiltration of mast cells in AA and AGA patients, and this increase was age and severity dependent. Moreover, the increase in mast cell proliferation is more dominant in AA patients. [GMJ.2020;9:e1962]


2006 ◽  
Vol 61 (5-6) ◽  
pp. 421-426 ◽  
Author(s):  
Dürdane Kolankaya ◽  
Hakan Şentürkb ◽  
Aslı Özkök Tüylü ◽  
Sibel Hayretdağ ◽  
Güldeniz Selmanoğlua ◽  
...  

Three kinds of pollen taxa belonging to 3 families (Fabaceae - Trifolium spp., Brassicaceae - Raphanus spp. and Cistaceae - Cistus spp.) and commonly collected by honeybees were fed to mature male rats separately, in the form of 60 mg/animal/day for a 30-day period. The objective of this study was to investigate any positive effects or possible side effects of the use of pollen on the immune system. This was achieved through blood analysis and cell count on blood, hemoglobin, erythrocyte and immune system cells. The cell concentration of mast cells, degranulization and cell localization were investigated in prepared mesentery tissue samples. Histological investigations of the stomach and duedenum sections of pollen-fed rats were carried out to learn the reason for eosinophil gastroenteritis in the alimentary canal. The eosinophil and lymphocyte levels of rats fed with pollen of Trifolium spp., Raphanus spp., and Cistus spp. were observed to have increased blood cell counts, while neutrophil and monocyte levels decreased; different values were found in basophil leucocytes between the pollen groups. Differing reductions in mesentery mast cell concentration, degranulization and cell localization were found. Within the three separate pollens, the rats having been fed with Cistus spp. pollen were observed to have higher blood lymphocyte, eosinophil, hemoglobin and hematocrit values than those fed with the others, as well as low mesentery mast cell concentration. Hemoglobin values were determined to increase at a proportion of between 10.0-11.3%. No difference was found in other blood parameters. The fat proportion of the male rats fed with the three taxa was between 4.03-8.75%, while that for protein proportion was between 16.11-24.25%. Male rats receiving these taxa did not experience allergic reactions and it is possible to argue that the low protein and fat content of these pollens have a strengthening effect on the immune systems by the increase in lymphocyte content and the amount of hemoglobin leads to an increase of oxygen transport capacity in the tissues.


1997 ◽  
Vol 11 (1) ◽  
pp. 63-66 ◽  
Author(s):  
Gilead Berger ◽  
Arnon Goldberg ◽  
Dov Ophir

The number of mast cells in the inferior turbinates of patients with perennial allergic rhinitis and perennial nonallergic rhinitis was compared with normal controls. Mast cell counts expressed as the mean number in 100 high-power fields, assessed after Carnoy's fixation and toluidine blue staining were 1.84 in normal controls (n = 11), 4.39 in patients with perennial allergic rhinitis (n = 13), and 4.00 in those with perennial nonallergic rhinitis (n = 26). Statistical analysis confirmed that the density of mast cells in allergic as well as in nonallergic patients was significantly higher than in normal controls, whereas no significant difference was found between the number of mast cells in allergic and nonallergic patients. It is concluded that the number of mast cells in the inferior turbinate mucosa of patients with perennial rhinitis is increased compared with normal controls, and the increased number is not necessarily allergy-dependent.


2018 ◽  
Vol 48 (9) ◽  
Author(s):  
Bárbara Hess Rodrigues Gonçalves ◽  
Bruna Dantas Matos ◽  
Mariana Batista Rodrigues Faleiro ◽  
Emmanuel Arnhold ◽  
Moema Pacheco Chediak Matos ◽  
...  

ABSTRACT: In this study the correlation between the clinical score, mast cell count and interleukin 31 (IL-31) immunostaining in the skin of dogs with atopic dermatitis was determined. A total of 31 dogs of different breeds, from one to eight years of age, were chosen for the study. The 20 females and 11 males were categorized based on the CADESI-4 system, as having discrete, moderate or marked atopic dermatitis. Skin samples were collected from the axillary and interdigital regions and stained with hematoxylin and eosin for cytohistomorphological analyses and toluidine blue to evaluate the mast cell counts, and immunohistochemistry for the IL-31 immunostaining. Animals revealing higher atopic dermatitis scores had greater numbers of mast cells and IL-31 immunolabeled cells. More numbers of cells immunolabeled for IL-31 were evident in the axillary skin compared with the interdigital skin in dogs having this condition. A correlation was identified between the clinical scores and mast cell numbers in the interdigital region, as well as between the clinical scores and number of cells immunolabeled for IL-31 in the axillary area. A correlation was also reported between the mast cell numbers and IL-31 immunolabeled cells only in the axillary skin, and none in the interdigital regions. It was thus concluded that the mast cells and IL-31 are involved in the pathogenesis of the canine atopic dermatitis (CAD), as well as lymphocytes and plasma cells. It was also observed that the higher the degree of clinical severity of the disease, the more the numbers of mast cells and IL-31 in the skin of those animals suffering from CAD, which implies the influence of these immunological constituents on the genesis of pruritus and disease progression.


1993 ◽  
Vol 107 (5) ◽  
pp. 418-422 ◽  
Author(s):  
Hirokuni Otsuka ◽  
Kimihiro Ohkubo ◽  
Harumi Seki ◽  
Masaki Ohnishi ◽  
Terumichi Fujikura

The distribution and abundance of mast cells in nasal polyps, the maxillary sinus mucosa of patients with sinusitis and the turbinate mucosa of allergic rhinitis was microscopically examined using different methods of fixation. In the epithelium of the surface and the ducts of nasal polyps (n = 8), the mean number of mast cells was over 20,000 per mm3 using Mota's fixation and the increase was correlated with the epithelial thickness (P<0.05). On the other hand those of the maxillary sinus mucosa (n = 6) and the nasal turbinate mucosa (n = 7) were less than 6,000 per mm3. In the subepithelial layer or areas deeper than the area with the glands, however, mast cell counts were less than 3,200 per mm3 in all diseases. More than 70–90 per cent of all mast cells in the epithelium of the mucosal surface and the ducts of the polyp, the maxillary sinus mucosa and nasal turbinates were formalin sensitive. Most of the mast cells in the subepithelial and deeper areas were formalin resistant in all diseases.These results suggest that conditions for mast cell growth differ between polyps and the other diseases, and that the conditions which affect mast cells may contribute to polyp development.


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