Potassium bromate (KBrO3) Modulates Oxidative stress and inflammatory biomarkers in NaOH induced crohn’s colitis in Wistar rats

Potassium bromate (KBrO3) present in consumed ozonised water was recently documented to exacerbate experimental gastric ulcer. Report is however vague as regards its effects in the colon; where water reabsorption occurs. In this study, we observed the possible effects of KBrO3 on oxidative stress and inflammatory biomarkers in NaOH induced crohn’s colitis. Wistar rats (180-200g) were divided into 6 groups (n=10); 1-control, 2-untreated crohn’s colitis (induced by 1.4% NaOH; intra-rectal administration) and 3-6: crohn’s colitis treated with vitamin E, KBrO3, vitamin E+KBrO3 and sulphazalazine respectively for seven days. Body weight and stool score were monitored daily. By 3 and 7, excised colon was evaluated for ulcer scores, biochemical and histological analysis. Collected blood samples on days 3 and 7 were assayed for haematological indices using standard methods. Data were subjected to ANOVA and p ≤0.05 considered significant. Platelet/lymphocyte ratio, colonic ulcer score, , malondialdehyde and mast cells were significantly decreased while colonic sulfhydryl, Ca2+ and Na+/K+ ATPase activities were increased following KBrO3 treatment compared with crohn’s colitis untreated. Findings suggest that KBrO3 may mitigate against NaOH induced crohn’s colitis via inhibiting mast cell population, oxidative and inflammatory content but stimulating colonic sulfhydryl, Ca2+ and Na+/K+ ATPase activities.

Tryptase Profile of the Rat Skin Mast Cell Population During the Wound Healing

Mast cells cyclically synthesize and excrete a wide range of biogenesis products with different biological activities into the extracellular matrix and are regulators of local homeostasis both in normal conditions and in pathology – inflammation, oncogenesis, etc. The relative specificity of classical histochemical methods for detecting mast cells in relation to chromogenic to substrates causes certain difficulties in the selective study of the components of the secretome of mast cells, for example, heparin, histamine, chymase or tryptase. Therefore, immunomorphological techniques have become very popular, which identify specific substrates and allow differentiation of the components of the mast cell secretome. Mediators produced by mast cells promote neoangiogenesis, fibrillogenesis and re-epithelialization during the repair process.The aim of our work was to study the tryptase profile of the mast cell population of rat skin during the wound processusing an original combined method of immunohistochemical staining.Material and methods. The experiment involved 12 Wistar rats divided into two groups – intact (n=6) and with the existing wound process of the skin in the withers (n=6). The tryptase profile of mast cells was assessed on the 7th day of the wound process in comparison with the control group.Results. The results obtained showed a significant increase in the number of tryptase-positive mast cells on the 7th day of the wound process in the skin against the background of a general increase in the population of mast cells. Intragranular tryptase reserve was significantly increased. In contrast to the control, where mast cells with single tryptase-positive granules dominated, during the wound process, cells of this type were practically not detected in the skin (43.69±2.9% and 8.55±0.9%). The content of tryptase-positive mast cells with complete filling of the cytoplasm in the control group and the group of animals with a wound process was 14.24±1.2% and 38.03±2.9%, respectively.Conclusion. Thus, when modeling a wound, an increase in tryptase synthesis is detected both in individual MCs and within the entire MC population. This fact indicates that mast cell proteases can become a potential therapeutic target for improving wound regeneration by correcting immunogenesis, inflammation and fiber formation.

Novel histochemical approach for evaluation of tryptase expression in the mast cell population

The paper presents a novel histochemical approach for evaluation of tryptase expression in the mast cell population. To study the selective effect of mast cells (MC) on the parameters of a specific tissue microenvironment, it is necessary to detail the molecular composition of their secretome and analyze the pathways of degranulation. The developed method for combined immunomorphological and histochemical tryptase staining protocols contributes to a more objective determination of the integral level of specific protease expression in the skin MC population. Extra visualization potentials of cytological features and specific aspects of tryptase processing expand efficiency of morphological analysis in both normal and pathological conditions.

Flow Cytometry-Based Characterization of Mast Cells in Human Atherosclerosis

The presence of mast cells in human atherosclerotic plaques has been associated with adverse cardiovascular events. Mast cell activation, through the classical antigen sensitized-IgE binding to their characteristic Fcε-receptor, causes the release of their cytoplasmic granules. These granules are filled with neutral proteases such as tryptase, but also with histamine and pro-inflammatory mediators. Mast cells accumulate in high numbers within human atherosclerotic tissue, particularly in the shoulder region of the plaque. These findings are largely based on immunohistochemistry, which does not allow for the extensive characterization of these mast cells and of the local mast cell activation mechanisms. In this study, we thus aimed to develop a new flow-cytometry based methodology in order to analyze mast cells in human atherosclerosis. We enzymatically digested 22 human plaque samples, collected after femoral and carotid endarterectomy surgery, after which we prepared a single cell suspension for flow cytometry. We were able to identify a specific mast cell population expressing both CD117 and the FcεR, and observed that most of the intraplaque mast cells were activated based on their CD63 protein expression. Furthermore, most of the activated mast cells had IgE fragments bound on their surface, while another fraction showed IgE-independent activation. In conclusion, we are able to distinguish a clear mast cell population in human atherosclerotic plaques, and this study establishes a strong relationship between the presence of IgE and the activation of mast cells in advanced atherosclerosis. Our data pave the way for potential therapeutic intervention through targeting IgE-mediated actions in human atherosclerosis.

MDS with 5q deletion and rare cKIT positive mastocytosis: a diagnostic and therapeutic challenge

A patient with a diagnosis of myelodysplastic syndrome (MDS) with isolated 5q deletion underwent repeat bone marrow biopsy to assess haematological response after 6 months of initial lenalidomide therapy. Subsequent bone marrow biopsies revealed persistent MDS with del(5q) in addition to a small atypical mast cell population with >25% of mast cells with spindle-shaped morphology and immunohistochemistry characteristics consistent with mastocytosis. Molecular testing on the bone marrow was positive for cKIT D816V and the patient was diagnosed with systemic mastocytosis (SM) with an associated haematological neoplasm. MDS with SM is well known to be associated; however, to the best of our knowledge, only one prior case report identifies MDS with del(5q) and associated cKIT D816V positive mastocytosis. While the exact clonal origin of both chromosomal aberrations is unclear, this case illustrates the therapeutic efficacy of lenalidomide in a patient with MDS with del(5q) and rarely associated cKIT positive SM.

The effect of vitamin E on mast cells in small intestine of broilers under heat stress

The aim of this study is to identify the effect of vitamin E (DL-α-tocopherol acetate) (300 IU/kg) on mast cells in the small intestine (duodenum, jejunum and ileum) under heat stress. In the study, 42 one-day-old Ross 308 male broiler chicks were used. The chicks were randomly separated into 3 groups as follows; control (22±2°C), heat stress (35°C, 5 hours/per day) and vitamin E (300 IU/kg/per day) + heat stress (35°C, 5 hours/per day). The applications of heat stress and vitamin E began on the fifteenth day and ended on the thirty-fifth day. Tissue samples were taken from animals in each group of four and five-week-old chickens. Tissue samples were fixed in BLA (Basic Lead Acetate) solution. The sections were stained with toluidine blue (TB) (pH 0.5) and alcian blue-critical electrolyte concentration (AB-CEC) (pH 5.8, 0.3 M MgCl2) / Safranin O (SO) (pH 1.0) combined method. It was determined that increasing of the exposure duration to heat stress increased the number of mast cells in the small intestine of the boilers. Also, it was revealed that vitamin E reduced mast cell population under heat stress. Consequently, heat stress may play a role in the pathogenesis of small intestine-associated with disorders and the supplementation of vitamin E can contribute to regulate small intestine functions of broilers by decreasing mast cell proliferation and activation under heat stress.