Mast Cell Tryptase Expression Research Articles

Expression of mast cell tryptase and immunoglobulin E is increased in cutaneous photodamage: implications for carcinogenesis

Purpose: Mast cells, their serine proteinase tryptase, and immunoglobulin E (IgE) can be involved in cutaneous carcinogenesis. Materials and methods: To study the association of tryptase+ and IgE+ cells with photodamage and skin cancers 385 adult patients (201 males, 184 females, 75 with immunosuppression) at risk of any type of skin cancer were examined. Skin biopsies were taken from the sun-protected medial arm and from the photodamaged dorsal forearm skin followed by immunohistochemical staining for tryptase and IgE. Results: The results show that tryptase+ and IgE+ cells are significantly higher in number in the photodamaged than sun-protected skin, both in immunocompetent and -compromised subjects, and there is a strong correlation between tryptase+ and IgE+ cells. The numbers of forearm tryptase+ and especially IgE+ cells associated significantly with the forearm photodamage severity. In the logistic regression analysis, the forearm to upper arm ratio of IgE+ cells produced a univariate odds ratio of 1.521 (p = .010) and a multivariate one of 3.875 (p = .047) for the history of squamous cell carcinoma. The serum level of total IgE correlated significantly to the IgE to tryptase ratio in both skin sites. Conclusions: Therefore, IgE+ mast cells participate in photodamage and carcinogenesis, though it is unclear whether they are tumor-protective or -causative.

Inhibition of vascular endothelial growth factor-A downregulates angiogenesis in psoriasis: A pilot study.

Vascular Endothelial Growth Factor (VEGF)-A-mediated angiogenesis participates in the pathogenesis of psoriasis, thus inviting the hypothesis that anti-VEGF-A therapy could be beneficial in psoriasis. While anti-angiogenic agents are used in oncology and ophthalmology, these therapeutic strategies remain unexplored for the management of psoriasis. Our objective was to investigate ex vivo how VEGF-A blockade impacts blood vessels, epidermis and immune cells in organ-cultured plaque and non-lesional skin from patients with psoriasis. Skin biopsies from patients with psoriasis (n=6; plaque and non-lesional skin) and healthy controls (n=6) were incubated with anti-VEGF-A monoclonal antibody (bevacizumab, Avastin®) or a human IgG1 isotype control for 72-h in serum-free organ culture. CD31/LYVE-1, Ki-67, and mast cell tryptase expression were assessed by quantitative immunohistomorphometry. VEGF-A levels in plasma, PBMCs and skin culture supernatants were measured. Inhibition of VEGF-A blocked all free VEGF-A ex vivo, reduced blood vessel area and the number of blood vessel endothelial cells in plaques of psoriasis (*p<0.05). The treatment effect correlated significantly with levels of VEGF-A in organ culture supernatants (r=0.94; *p<0.05) from plaque skin and with plasma levels of VEGF-A from patients with psoriasis (r=0.943; *p=0.017). These ex vivo data are the first studies to objectively investigate the potential of VEGF-A inhibition as a novel adjuvant treatment strategy for psoriasis. Taken together, our data encourage further investigation by clinical trial to explore whether downregulating pathological angiogenesis has clinical utility, especially in patients with severe psoriasis or those with elevated levels of VEGF-A in plasma and/or skin.

Involvement of Mast-Cell-Tryptase- and Protease-Activated Receptor 2-Mediated Signaling and Urothelial Barrier Dysfunction with Reduced Uroplakin II Expression in Bladder Hyperactivity Induced by Chronic Bladder Ischemia in the Rat.

We aimed to investigate the relationship between mast cell (MC) infiltration into the bladder with urothelial barrier dysfunction and bladder hyperactivity in a chronic bladder ischemia (CBI) rat model. We compared CBI rats (CBI group; n = 10) with normal rats (control group; n = 10). We measured the expression of mast cell tryptase (MCT) and protease-activated receptor 2 (PAR2), which are correlated with C fiber activation via MCT, and Uroplakins (UP Ia, Ib, II and III), which are critical to urothelial barrier function, via Western blotting. The effects of FSLLRY-NH2, a PAR2 antagonist, administered intravenously, on the bladder function of CBI rats were evaluated with a cystometrogram. In the CBI group, the MC number in the bladder was significantly greater (p = 0.03), and the expression of MCT (p = 0.02) and PAR2 (p = 0.02) was significantly increased compared to that of the control group. The 10 μg/kg FSLLRY-NH2 injection significantly increased the micturition interval of CBI rats (p = 0.03). The percentage of UP-II-positive cells on the urothelium with immunohistochemical staining was significantly lower in the CBI group than in the control group (p < 0.01). Chronic ischemia induces urothelial barrier dysfunction via impairing UP II, consequently inducing MC infiltration into the bladder wall and increased PAR2 expression. PAR2 activation by MCT may contribute to bladder hyperactivity.

MP49-16 IMPAIRED UROTHELIAL BARRIER FUNCTION INDUCED LOWER URINARY TRACT DYSFUNCTION BY PROTEASE-ACTIVATED RECEPTER 2 ACTIVATION

You have accessJournal of UrologyCME1 May 2022MP49-16 IMPAIRED UROTHELIAL BARRIER FUNCTION INDUCED LOWER URINARY TRACT DYSFUNCTION BY PROTEASE-ACTIVATED RECEPTER 2 ACTIVATION Hidenori Akaihata, Yusuke Hakozaki, Hiroki Natsuya, Tomoyuki Kumekawa, Shunsuke Yoshioka, Hitomi Imai, Akifumi Onagi, Kanako Matsuoka, Tomoyuki Koguchi, Junya Hata, Yuichi Sato, Masao Kataoka, and Yoshiyuki Kojima Hidenori AkaihataHidenori Akaihata More articles by this author , Yusuke HakozakiYusuke Hakozaki More articles by this author , Hiroki NatsuyaHiroki Natsuya More articles by this author , Tomoyuki KumekawaTomoyuki Kumekawa More articles by this author , Shunsuke YoshiokaShunsuke Yoshioka More articles by this author , Hitomi ImaiHitomi Imai More articles by this author , Akifumi OnagiAkifumi Onagi More articles by this author , Kanako MatsuokaKanako Matsuoka More articles by this author , Tomoyuki KoguchiTomoyuki Koguchi More articles by this author , Junya HataJunya Hata More articles by this author , Yuichi SatoYuichi Sato More articles by this author , Masao KataokaMasao Kataoka More articles by this author , and Yoshiyuki KojimaYoshiyuki Kojima More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002624.16AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Recently, attention has focused on the association between urothelial barrier dysfunction and lower urinary tract dysfunction (LUTD). Uroplakin (UP) is critical to normal urothelial barrier function. Some studies reported that UPs defects lead to abnormal voiding patterns. It was also reported that impairing UP II causes mast cell infiltration in the bladder and that mast cell contributed to bladder hyperactivity. Mast cell tryptase (MCT) can regulate neuronal activity by cleaving protease-activated recepter 2 (PAR2), which is expressed on C-fiber. On the other hands, the mechanisms underlying LUTD caused by chronic ischemia are not completely known. This study aimed to investigate the effects of chronic ischemia on UPs and mast cell in the bladder using a rat model of chronic bladder ischemia (CBI). METHODS: Adult male Sprague-Dawley rats (16 weeks old) were divided into two groups (control and CBI; n=10 each). The CBI group received balloon endothelial injury of bilateral iliac arteries and a 2% cholesterol diet for 8 weeks to induce chronic ischemia. The control group received regular diet for 8 weeks. After monitoring urine output for 24 h, bladders were harvested. Bladder was processed for immunohistochemical staining and methylene blue staining. We used western blotting to measure expressions of UPs(UP Ia, UP Ib, UP II, UP III) in the bladder of this rat model to examine urothelial barrier function. The expression of MCT and PAR2 were also measured to evaluate C-fiber activation by mast cell. RESULTS: Metabolic cage studies showed that mean voided volumes were significantly smaller in the CBI (1.46±0.33 ml) than in the control (1.01±0.21 ml; P=0.001). In western blot analysis, UP II expression was significantly decreased (P=0.011) in the CBI compared with the control. With immunohistochemical staining, UP II-positive cells are located mostly. The percentage of UP II-positive cells on urothelium was significantly lower in the CBI (63.0±0.11%) than in the control (92.1±0.1%, P<0.001). The expression of MCT (P=0.024) and PAR2 (P=0.021) were significantly increased in the CBI compared with the control in western blot analysis. The methylene blue staining showed the number of mast cell was significantly more in the CBI than in the control (P=0.034). CONCLUSIONS: Chronic ischemia might impair urothelial barrier function through reduced expression of UP II in urothelium. Urothelial barrier dysfunction contributed to mast cell infiltration and increased PAR2 on C-fiber, resulting PAR2 activation by MCT. The PAR2 activation may induce LUTD by stimulating C fiber in rat model of chronic bladder ischemia. Source of Funding: None © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e860 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Hidenori Akaihata More articles by this author Yusuke Hakozaki More articles by this author Hiroki Natsuya More articles by this author Tomoyuki Kumekawa More articles by this author Shunsuke Yoshioka More articles by this author Hitomi Imai More articles by this author Akifumi Onagi More articles by this author Kanako Matsuoka More articles by this author Tomoyuki Koguchi More articles by this author Junya Hata More articles by this author Yuichi Sato More articles by this author Masao Kataoka More articles by this author Yoshiyuki Kojima More articles by this author Expand All Advertisement PDF DownloadLoading ...

Electroacupuncture Inhibits Inflammatory Response in Rats With Interstitial Cystitis

OBJECTIVE: Electroacupuncture (EA) has been clinically utilized to manage interstitial cystitis (IC), but its related mechanism research is remained to be researched. This study aimed to explore the role of EA in inhibiting the inflammatory response of IC rats, and research the possible mechanism based on the changes of urodynamics, mast cell tryptase expression and the content of inflammatory substances.&lt;br/&gt; METHODS: EA (continuous wave, 30 Hz, 1 mA) was applied to stimulate the Ciliao point (BL32) and the Huiyang point (BL35) of rats. Changes of urodynamics, mast cell tryptase expression and the content of IL-6 and TNF-α were detected after 24 hours of the treatment.&lt;br/&gt; RESULTS: The results showed that after EA treatment, compared with the model group, the initial urination time of the rats was prolonged (3.23 ± 0.60 to 5.27 ± 0.27 min), the bladder capacity was increased (0.80 ± 0.15 to 1.31 ± 0.07 ml), and the degree of bladder mucosal damage was restored. The expression of mast cell tryptase, and the content of IL-6 and TNF-α in its bladder tissue and urine were reduced.&lt;br/&gt; CONCLUSION: These findings suggested that EA could suppress bladder overactivity and inhibit inflammatory response of IC rats, and its mechanism was proved to be related to the inhibition of mast cell degranulation and the down-regulation of inflammatory substances IL-6 and TNF-α in bladder tissue.

Mast cells in luminal vs non-luminal breast cancers

Background: Tumor growth and development is determined by the mutual interaction between the cancer cells themselves and the microenvironment. It contains various elements, including immune cells. Of all, mast cells have one of the most controversial roles. The aim of the present study was to evaluate the expression of mast cell tryptase in the luminal and non-luminal subtypes of breast cancer and establish a possible link between infiltration with MCs and expression of hormone receptors. Material and methods: The experimental study included 80 cases of breast carcinomas that were analyzed immunohistochemically in order to establish the molecular profile and the expression of tryptase, a specific marker of mast cells. The data were processed using the SPSS program. Pearson’s coefficient (r) and the other values were considered statistically significant in case of p≤0.05. Results: Both intratumoral mast cells (MCit) and peritumoral mast cells (MCpt) correlated with the expression of hormone receptors for estrogen (ER) and progesterone (PR). Thus, the following relations were established: MCit and ER (r=0.343, p=0.002), MCpt and ER (r=0.394, p=0.000295) and MCpt and PR (r=0.386, p=0.000409). Statistically significant correlations between HER2 expression and mast cells content have not been established. Conclusions: Mast cells invasion, peri- and intratumoral, is strongly influenced by the expression of hormone receptors. The luminal subtypes of breast cancer are characterized by a higher density of mast cells.

Stromal infiltrating mast cells identify immunoevasive subtype high-grade serous ovarian cancer with poor prognosis and inferior immunotherapeutic response

ABSTRACT Tumor infiltrating mast cells (TIMs), with pro- or anti-tumorigenic role in different types of malignancies, have been implicated in resistance to anti-PD1 therapy. Here, we aimed to identify the relevance of TIMs with the prognosis, immune contexture, and immunotherapy in high-grade serous ovarian cancer (HGSOC). Tissue microarrays containing 197 HGSOC patients were assessed by immunohistochemistry (IHC) for detecting the expression of mast cell tryptase and other immune markers. Kaplan-Meier curve, log-rank test, and Cox regression model were applied to perform survival analysis. Single-cell RNA-seq analysis and flow cytometric analysis were selected to characterize TIMs. Furthermore, short-term HGSOC organoids were employed to validate the effect of TIMs on anti-PD1 therapy. Abundance of stromal TIMs (sTIMs) predicted dismal prognosis and linked to immunoevasive subtype of HGSOC, characterized by increased infiltration of pro-tumor cells (Treg cells, M2-polarized macrophages, and neutrophils) and impaired anti-tumor immune functions. Intensive inter-cell interactions between TIMs and other immune cells were identified, suggesting potential cross-talks to foster an immunosuppressive microenvironment. Organoids derived from sTIMs-low patients were associated with increased response to anti-PD-1 treatment other than the presence of high sTIMs infiltration. A nomogram, constructed by combining FIGO stage, sTIMs, and PD-L1, with an area under the curve (AUC) for predicting 5-year overall survival of 0.771 was better than that of FIGO staging system of 0.619. sTIMs/PD-L1-based classifier has potential clinical application in predicting prognosis of patients with HGSOC. sTIMs-high tumors correlate with immunosuppressive tumor microenvironment (TME) and possess potential insensitivity to immunotherapy.

Tespa1 plays a role in the modulation of airway hyperreactivity through the IL-4/STAT6 pathway

BackgroundThymocyte-expressed, positive selection-associated 1 (Tespa1) is a critical signaling molecule in thymocyte development. This study aimed to investigate the regulatory effect of Tespa1 on mast cells in the pathogenesis of asthma and its relationship with the interleukin (IL)-4/signal transducers and activators of transcription 6 (STAT6) signaling pathway.MethodsTespa1 mRNA expression analysis and IgE levels were carried out using the induced sputum of 33 adults with stable asthma and 36 healthy controls. Tespa1-knockout mice (Tespa1−/−, KO) and C57BL/6 background (wild-type, WT) mice were sensitized and treated with ovalbumin (OVA) to establish an asthma model. Pathological changes, number and activity of mast cells, and changes in activation of the IL-4/STAT6 pathway in lung tissue were detected. The changes of tryptase expression and STAT6 activation after mast cell gene knockout were analyzed in vitro. The changes of enzyme expression and STAT6 activation after mast cell gene knockout were analyzed in vitro. The association between the Tespa1 and p-STAT6 was analyzed by co-immunoprecipitation method.ResultsCompared with the healthy controls, Tespa1 expression was decreased, and IgE levels were elevated in the sputum of asthmatic patients. Animal experiments showed that Tespa1−/− mice exhibited more severe inflammation, higher quantity of goblet cells and mast cells in the bronchium, and greater expression of mast cell tryptase, which is induced by ovalbumin, than WT mice. And IL-4, IL-13, phospho-Janus kinase 1, and p-STAT6 expressions presented a higher increase in the Tespa1−/− mouse model than in the WT mouse model. Further in vitro studies confirmed that IL-4 could more significantly promote tryptase and p-STAT6 activities in Tespa1−/− mast cells than their WT counterparts. Correlation analysis results showed a negative correlation between Tespa1 and p-STAT6. Co-immunoprecipitation results demonstrated an association between Tespa1 and p-STAT6.ConclusionsAltogether, our results indicate that Tespa1 can negatively regulate mast cell activity, and this event is related to the mast cell IL-4/STAT6 signaling pathway and could be therapeutically exploited to treat asthma attacks.

GPER‐mediated, oestrogen‐dependent visceral hypersensitivity in stressed rats is associated with mast cell tryptase and histamine expression

Visceral hypersensitivity (VH) is common in irritable bowel syndrome (IBS), and female patients are more likely to seek healthcare services for IBS-related abdominal pain. Oestrogen has been reported to mediate pain modulation via its receptor, and mast cells are known to participate in the development of visceral hypersensitivity. Our previous studies showed that the G-protein-coupled oestrogen receptor (GPER, also known as GPR30) was expressed by mast cells in human colonic tissues and was associated with IBS type and severity of visceral pain. However, whether GPER is involved in oestrogen-dependent visceral hypersensitivity via mast cell degranulation is still unknown. Rats were subjected to wrap partial restraint stress to induce visceral hypersensitivity and were ovariectomized (OVX) to eliminate the effects of oestrogen on visceral hypersensitivity. OVX rats were treated with oestrogen, an oestrogen receptor α and β antagonist (ICI 182.780), a GPER antagonist (G15) or a GPER agonist (G1), to evaluate the effects of oestrogen via its receptor. The colorectal distention test was performed to assess visceral sensitivity. Immunofluorescence studies were performed to evaluate GPER and mast cell tryptase co-expression. Mast cell number with degranulation was detected by specific staining. Mast cell tryptase expression in rat colon was also investigated by Western blot and immunohistochemistry. Substance P and histamine expression were examined by ELISA. GPER was expressed by the majority of tryptase-positive mast cells in the colonic mucosa. Stressed rats showed increased visceral sensitivity, increased mast cell degranulation, mast cell tryptase expression, and increased colon histamine levels. Ovariectomy reduced stress-induced VH in female rats and decreased mast cell degranulation, mast cell tryptase expression, and histamine levels, whereas oestrogen replacement reversed these effects. In OVX rats, the GPER antagonist G15 counteracted the enhancing effects of oestrogen on stress-induced VH, mast cell degranulation, mast cell tryptase, and histamine expression, whereas VH was preserved after treatment with ICI 182.780. On the other hand, pretreatment with the selective GPER agonist G1 at doses between 1 and 20μg/kg significantly increased VH, mast cell tryptase, and histamine expression in OVX-stressed rats, mimicking the effects of oestrogen. GPER plays a pivotal role in the regulation of mast cell degranulation, mast cell tryptase expression, and histamine levels and contributes to the development of colonic hypersensitivity in a female rat model of IBS.

Angiogenesis in bladder tissues is strongly correlated with urinary frequency and bladder pain in patients with interstitial cystitis/bladder pain syndrome.

To examine the correlation among bladder inflammation, angiogenesis, fibrosis and urothelial denudation in biopsied bladder specimens, and O'Leary-Sant symptom indexes, O'Leary-Sant problem indexes and visual analog scale pain scores in interstitial cystitis/bladder pain syndrome patients with or without Hunner lesions (Hunner type interstitial cystitis or non-Hunner type interstitial cystitis). Bladder biopsied tissues were collected from 12 Hunner type interstitial cystitis female patients, 12 non-Hunner type interstitial cystitis female patients and 12 age-matched non-interstitial cystitis female patients (controls). Immunohistochemical stainings of tissue necrotic factor-α, mast cell tryptase, vascular endothelial growth factor, CD31, transforming growth factor-β, SLUG associated with epithelial mesenchymal transition and E-cadherin as well as Masson trichrome staining were evaluated. The significant correlation between the expression of tissue necrotic factor-α, mast cell tryptase, vascular endothelial growth factor, CD31, transforming growth factor-β, collagen, SLUG or E-cadherin, and O'Leary-Sant symptom indexes, O'Leary-Sant problem indexes or visual analog scale pain scores was then examined. The expression of tissue necrotic factor-α, vascular endothelial growth factor, CD31, transforming growth factor-β and SLUG was significantly increased in non-Hunner type interstitial cystitis and Hunner type interstitial cystitis patients compared with controls whereas the significant increases in the expression of mast cell tryptase and collagen were observed in Hunner type interstitial cystitis patients compared with controls and non-Hunner type interstitial cystitis patients. On the other hand, the expression of E-cadherin was significantly decreased in Hunner type interstitial cystitis patients compared with controls and non-Hunner type interstitial cystitis patients. In addition, the increased expression of CD31 in bladder tissues was strongly correlated with O'Leary-Sant symptom indexes, O'Leary-Sant problem indexes and visual analog scale pain scores. These results suggest that bladder angiogenesis evident as the increased expression of CD31 is strongly correlated with urinary frequency and bladder pain in patients with non-Hunner type interstitial cystitis and Hunner type interstitial cystitis.

The role of toll-like receptor 4 and mast cell in the ameliorating effect of electroacupuncture on visceral hypersensitivity in rats.

Visceral hypersensitivity is one of the main pathogenesis of irritable bowel syndrome (IBS) and mast cell activation is closely related to visceral hypersensitivity. As a critical molecule of the innate immune system, toll-like receptor 4 (TLR4) may modulate the activation of mast cell. Alleviating effect of electroacupuncture (EA) on visceral hypersensitivity has been proved, whereas, whether the TLR4 and mast cell is involved in this process remains unclear. Forty Sprague-Dawley rats were randomly divided into five groups: control group, model group, EA group, sham EA group, and mast cell stabilizer (MCS) group. Visceral sensitivity during colorectal distension was assessed by the measurement of visceral motor reflex (VMR). TLR4 mRNA and protein expression were assessed by real-time PCR and immunohistochemistry, respectively. Mast cell number and mast cell tryptase (MCT) expression were detected. The level of inflammatory cytokine in serum was detected with ELISA. Visceral sensitivity was significantly higher in the model group than in the control group. EA and MCS significantly reduced VMR score at 0.8mL and 1.2mL distention pressures. Compared with model group, TLR4 mRNA expression, the protein expression of TLR4 and MCT, and the number of mast cells with degranulation in the colonic tissue, serum concentration of IL-1β and IL-8 were all significantly decreased in EA and MCS group. Electroacupuncture ameliorated visceral hypersensitivity in colon-sensitized model probably via decreasing the level of pro-inflammatory cytokines released by mast cell which were decreased when the TLR4 expression in the colonic tissue was downregulated by EA.

Alginate oligosaccharide enhances intestinal integrity of weaned pigs through altering intestinal inflammatory responses and antioxidant status.

Alginate oligosaccharide (AOS), prepared from depolymerised alginate, a natural polysaccharide occurring in the cell walls of brown algae, provides beneficial effects for intestinal health. However, the underlying mechanisms by which AOS supplementation maintains the intestinal integrity of weaned pigs remain obscure. Here, we aimed to determine how AOS modulates the intestinal integrity of weaned pigs. Twenty-four weaned pigs were assigned to two treatments: a control group (basal diet) and an AOS group (the basal diet supplemented with 100 mg kg−1 AOS). On day 15, eight pigs per treatment were randomly selected and sacrificed for serum and intestinal samples. We observed that AOS supplementation enhanced the intestinal integrity, as evidenced by the increased (P < 0.05) intestinal occludin protein abundance. Compared to the control group, AOS ingestion both elevated (P < 0.05) the jejunal and ileal catalase activity and decreased (P < 0.05) the duodenal and jejunal tumour necrosis factor-α concentration and mast cell tryptase expression. Furthermore, AOS down-regulated (P < 0.05) the duodenal toll-like receptor 4 (TLR4) and its down-stream signals, myeloid differentiation factor 88 (MyD88), interleukin-1 receptor-associated kinase 1 (IRAK1) and tumour necrosis factor receptor-associated factor 6 (TRAF6) mRNA levels, as well as jejunal nucleotide-binding oligomerisation domain protein 1 (NOD1) and its adaptor molecule, receptor-interacting serine/threonine-protein kinase 2 (RIPK2), mRNA levels. Additionally, phospho-nuclear factor-κB (p-NF-κB) p65 protein abundance in the duodenum and jejunum was down-regulated (P < 0.05) following AOS supplementation. According to the above results, the enhanced intestinal integrity in AOS-supplemented pigs appears to be associated with the elevated antioxidant capacity and the reduced mast cell degranulation, as well as the inhibited pro-inflammatory cytokines production via inhibiting the TLR4/NF-κB and NOD1/NF-κB signalling pathways.

Stress induces more serious barrier dysfunction in follicle-associated epithelium than villus epithelium involving mast cells and protease-activated receptor-2

Psychological stress has been associated with intestinal epithelial hyperpermeability, the basic process in various functional and organic bowel diseases. In the present study, we aimed to clarify the differences and underlining mechanisms in stress-induced barrier disruption in functionally and structurally distinct epitheliums, including the villus epithelium (VE) and follicle-associated epithelium (FAE), a specialized epithelium overlaid the domes of Peyer’s lymphoid follicles. Employing an Ussing Chamber system, the epithelial permeability was assessed in rats following water avoidance stress (WAS) in vivo and in mucosa tissues exposed to corticotropin-releasing factor (CRF) ex vivo. Decreased transepithelial resistance (TER) and increased paracellular and transcellular macromolecular permeability in colon, ileal VE and FAE had been observed in WAS rats and in CRF-exposed mucosa. Especially, the barrier dysfunction was more serious in the FAE. Moreover, WAS upregulated the expression of mast cell tryptase and protease-activated receptor-2 (PAR2), which positively correlated with epithelial conductance. Mast cell stabilizer cromolyn sodium obviously alleviated the barrier disruption induced by WAS in vivo and CRF in vitro. Serine protease inhibitor aprotinin and FUT-175, and selective PAR2 antagonist ENMD-1068 effectively inhibited the CRF-induced FAE hyperpermeability. Altogether, it concluded that the FAE was more susceptible to stress, and the mast cells and PAR2 signaling played crucial roles in this process.

Combined Analysis of Vascular Endothelial Growth Factor Expression with Cyclooxygenase-2 and Mast Cell Density in Oral Squamous Cell Carcinoma

Background: Overexpression of vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) and increasing mast cell density (MCD) in premalignant and malignant oral lesions have been documented. However, their correlation with clinicopathologic parameters and survival rate in oral squamous cell carcinoma (OSCC) is not completely clear. This study aimed to assess these subjects. Methods: VEGF, COX-2, and mast cell tryptase expression were examined immunohistochemically in 57 cases of OSCC. The relationships between the markers' expression and clinicopathologic data were assessed using bivariate and multivariate analysis. Results: Spearman's rank correlation coefficient showed a significant correlation between VEGF and COX-2 expression (r = 0.462, p < 0.001), as well as between VEGF expression and MCD (r = 0.306, p < 0.001). Multivariate analysis showed no significant correlation between the markers' immunoexpression and overall survival (OS), but a significant correlation between mode of invasion and OS [hazard ratio 0.362 (95% CI: 0.138- 0.974); p = 0.038] was observed. An association between MCD and gender (p = 0.042) was also found, as MCD was higher in males. Conclusion: The significant correlation of VEGF expression with COX-2 expression and MCD may represent the roles of COX-2 and MCD in tumor angiogenesis by modulating VEGF production. However, VEGF, COX-2, and MCD are not useful indicators to predict prognosis in OSCC. Nevertheless, the mode of invasion can be considered as an independent prognostic factor in OSCC patients.

Expressions of Mast Cell Tryptase and Brain Natriuretic Peptide in Myocardium of Sudden Death due to Hypersensitivity and Coronary Atherosclerotic Heart Disease.

To explore the value of mast cell tryptase and brain natriuretic peptide（BNP） in the differential diagnostic of sudden death due to hypersensitivity and coronary atherosclerotic heart disease. Totally 30 myocardial samples were collected from the autopsy cases in the Department of Forensic Pathology, Shanxi Medical University during 2010-2015. All samples were divided into three groups： death of craniocerebral injury group, sudden death of hypersensitivity group and sudden death of coronary atherosclerotic heart disease group, 10 cases in each group. Mast cell tryptase and BNP in myocardium were detected by immunofluorescence staining and Western Blotting. Immunofluorescence staining showed that the positive staining mast cell tryptase appeared in myocardium of sudden death of hypersensitivity group and coronary atherosclerotic heart disease group. Among the three groups, the expression of mast cell tryptase showed significantly differences through pairwise comparison （P<0.05）; The expression level of BNP in sudden death of coronary atherosclerotic heart disease group were significantly higher than the sudden death of hypersensitivity group and death of craniocerebral injury group （P<0.05）. The difference of the expression level of BNP between the sudden death of hypersensitivity group and the death of craniocerebral injury group had no statistical significance （P>0.05）. The combined detection of the mast cell tryptase and BNP in myocardium is expected to provide help for the forensic differential diagnosis of sudden death due to hypersensitivity and coronary atherosclerotic heart disease.

Acute Stress-Induced Changes in Follicular Dermal Papilla Cells and Mobilization of Mast Cells: Implications for Hair Growth.

BackgroundStress is a known cause of hair loss in many species.ObjectiveIn this study, we investigated the role of acute stress on hair growth using a rat model.MethodsRats were immobilized for 24 hours and blood samples, and skin biopsies were taken. The effect of stress-serum on the in vitro proliferation of rat and human dermal papilla cells (hDPCs), as well as serum cortisol and corticotropin-releasing hormone levels, were measured. Mast cell staining was performed on the biopsied tissue. In addition, Western blot and quantitative real time polymerase chain reaction were used to assess mast cell tryptase and cytokine expression, respectively in rat skin biopsies.ResultsStress-serum treatment reduced significantly the number of viable hDPCs and arrested the cell cycle in the G1 phase, compared to serum from unrestrained rats (p<0.05, respectively). Moreover, restrained rats had significantly higher levels of cortisol in serum than unrestrained rats (p<0.01). Acute stress serum increased mast cell numbers and mast cell tryptase expression, as well as inducing interleukin (IL)-6 and IL-1β up-regulation.ConclusionThese results suggest that acute stress also has an inhibitory effect on hair growth via cortisol release in addition to substance P-mast cell pathway.

Mo1870 Measuring Anti-Vinculin and Anti-CdtB Antibodies in Two Animal Models of Post-Infectious IBS

Diverticulosis is a common GI condition among older adults in the United States. However, limited information exists on the etiologic risk factors for diverticulosis. We hypothesized that the gut microbiome and chronic inflammation contribute to the development of diverticulosis.We evaluated the association between gut bacteria, mucosal inflammation and diverticulosis in a case-control study. Methods:Participants (n=196) were consented patients who underwent screening colonoscopy at UNC Hospitals. Subjects were classified as case if diverticula were present or control if no diverticula were present. The abundance of specific bacteria in the sigmoid mucosa was determined by qPCR based on the bacterial 16s rRNA gene for Bacteroides, Bifidobacterium, Clostridium, E. coli, Lactobacillus, and Eubacteria.Mucosal mRNA gene expression of cytokines of IL-6, IL-10, IL-17, and TNF-α and housekeeping gene HMBS were assessed by qRT-PCR. Immune cells in the lamina propria were stained for CD4, CD8, CD57 and Mast cell tryptase (MCT) by immunohistochemistry (IHC). The IHC stained sections were digitally imaged using the Aperio ScanScope XT. Analysis of biomarkers was performed using Definiens Tissue Studio software with the Composer_ Nuclei_and_Simulated_Cells algorithm. The percent cells with strong (+3), medium (+2) and weak (+1) positive signal, was used to compare biomarker levels among tissue samples. Statistical analysis included t-test and logistic regression to compare cases and controls as well as Spearman correlation to assess association between bacteria and inflammation. Results:We observed a higher relative abundance of Clostridium, Bacteroides, Bifidobacterium, E. coli, and Eubacteria in cases compared to controls. Cases had lower density of CD8 compared to controls. Although the cytokine gene expression levels of IL-6, IL-10, IL-17, and TNF-α did not differ between cases and controls, there was a significant positive correlation between CD57 expression and IL-6 (r=0.23, p=0.03), CD57 and TNFα (r0.33, p=0.001) and CD57 and IL-10 (r=0.22, p= 0.03) among cases. The association between bacterial abundance and immune cell density also varied depending on case-control status. Among cases, MCT expression showed positive correlations with Bacteroides (r=0.21, p= 0.05), Clostridium (r=0.18, p=0.08) and inverse correlation with Bifidobacteria (r=-0.26, p= 0.01). Among controls, CD57 expression was inversely correlated with Lactobacillus (r=0.19, p=0.06). Conclusions: Our findings suggest that gut bacteria and inflammation are associated with diverticulosis. This data combined with further research could have clinical implications for the prevention and treatment of diverticulosis.

Mo1869 Gut Bacteria and Inflammation Are Associated With Diverticulosis

Diverticulosis is a common GI condition among older adults in the United States. However, limited information exists on the etiologic risk factors for diverticulosis. We hypothesized that the gut microbiome and chronic inflammation contribute to the development of diverticulosis.We evaluated the association between gut bacteria, mucosal inflammation and diverticulosis in a case-control study. Methods:Participants (n=196) were consented patients who underwent screening colonoscopy at UNC Hospitals. Subjects were classified as case if diverticula were present or control if no diverticula were present. The abundance of specific bacteria in the sigmoid mucosa was determined by qPCR based on the bacterial 16s rRNA gene for Bacteroides, Bifidobacterium, Clostridium, E. coli, Lactobacillus, and Eubacteria.Mucosal mRNA gene expression of cytokines of IL-6, IL-10, IL-17, and TNF-α and housekeeping gene HMBS were assessed by qRT-PCR. Immune cells in the lamina propria were stained for CD4, CD8, CD57 and Mast cell tryptase (MCT) by immunohistochemistry (IHC). The IHC stained sections were digitally imaged using the Aperio ScanScope XT. Analysis of biomarkers was performed using Definiens Tissue Studio software with the Composer_ Nuclei_and_Simulated_Cells algorithm. The percent cells with strong (+3), medium (+2) and weak (+1) positive signal, was used to compare biomarker levels among tissue samples. Statistical analysis included t-test and logistic regression to compare cases and controls as well as Spearman correlation to assess association between bacteria and inflammation. Results:We observed a higher relative abundance of Clostridium, Bacteroides, Bifidobacterium, E. coli, and Eubacteria in cases compared to controls. Cases had lower density of CD8 compared to controls. Although the cytokine gene expression levels of IL-6, IL-10, IL-17, and TNF-α did not differ between cases and controls, there was a significant positive correlation between CD57 expression and IL-6 (r=0.23, p=0.03), CD57 and TNFα (r0.33, p=0.001) and CD57 and IL-10 (r=0.22, p= 0.03) among cases. The association between bacterial abundance and immune cell density also varied depending on case-control status. Among cases, MCT expression showed positive correlations with Bacteroides (r=0.21, p= 0.05), Clostridium (r=0.18, p=0.08) and inverse correlation with Bifidobacteria (r=-0.26, p= 0.01). Among controls, CD57 expression was inversely correlated with Lactobacillus (r=0.19, p=0.06). Conclusions: Our findings suggest that gut bacteria and inflammation are associated with diverticulosis. This data combined with further research could have clinical implications for the prevention and treatment of diverticulosis.

Mast cell tryptase and carboxypeptidase A expression in body fluid and gastrointestinal tract associated with drug-related fatal anaphylaxis.

To investigate the expression of mast cell tryptase and carboxypeptidase A in drug-related fatal anaphylaxis. The expression of mast cell tryptase and carboxypeptidase A in 15 autopsy cases of drug-related fatal anaphylaxis and 20 normal autopsy cases were detected. First, the expression of mast cell tryptase was determined in stomach, jejunum, lung, heart, and larynx by immunofluorescence. Different tissues were removed and fixed in paraformaldehyde solution, then paraffin sections were prepared for immunofluorescence. Using specific mast cell tryptase and carboxypeptidase A antibodies, the expression of tryptase and carboxypeptidase A in gastroenterology tract and other tissues were observed using fluorescent microscopy. The postmortem serum and pericardial fluid were collected from drug-related fatal anaphylaxis and normal autopsy cases. The level of mast cell tryptase and carboxypeptidase A in postmortem serum and pericardial fluid were measured using fluor enzyme linked immunosorbent assay (FEIA) and enzyme linked immunosorbent assay (ELISA) assay. The expression of mast cell tryptase and carboxypeptidase A was analyzed in drug-related fatal anaphylaxis cases and compared to normal autopsy cases. The expression of carboxypeptidase A was less in the gastroenterology tract and other tissues from anaphylaxis-related death cadavers than normal controls. Immunofluorescence revealed that tryptase expression was significantly increased in multiple organs, especially the gastrointestinal tract, from anaphylaxis-related death cadavers compared to normal autopsy cases (46.67 ± 11.11 vs 4.88 ± 1.56 in stomach, 48.89 ± 11.02 vs 5.21 ± 1.34 in jejunum, 33.72 ± 5.76 vs 1.30 ± 1.02 in lung, 40.08 ± 7.56 vs 1.67 ± 1.03 in larynx, 7.11 ± 5.67 vs 1.10 ± 0.77 in heart, P < 0.05). Tryptase levels, as measured with FEIA, were significantly increased in both sera (43.50 ± 0.48 μg/L vs 5.40 ± 0.36 μg/L, P < 0.05) and pericardial fluid (28.64 ± 0.32 μg/L vs 4.60 ± 0.48 μg/L, P < 0.05) from the anaphylaxis group in comparison with the control group. As measured by ELISA, the concentration of carboxypeptidase A was also increased more than 2-fold in the anaphylaxis group compared to control (8.99 ± 3.91 ng/mL vs 3.25 ± 2.30 ng/mL in serum, 4.34 ± 2.41 ng/mL vs 1.43 ± 0.58 ng/mL in pericardial fluid, P < 0.05). Detection of both mast cell tryptase and carboxypeptidase A could improve the forensic identification of drug-related fatal anaphylaxis.

The role of mast cells and mast cell tryptase in skin tags: an immunohistochemical study

Objective The aim of this study was to assess mast cell tryptase (MCT) expression in skin tags (STs) in comparison with normal skin. Background STs or acrochordons are common benign connective tissue of the dermis that are composed of loose fibrous tissue, in middle-aged and elderly individuals, located predominantly in intertriginous skin and neck, which consists of a bit of skin projecting from the surrounding skin. Patients and methods This study was carried out on 30 patients and 10 control participants. One biopsy was obtained from each patient. Mast cell count and mast cell density were assessed by immunohistochemical staining for MCT and expressed as mean, median, and range (the average mast cell count/high-power field). Results The results of the present study showed a statistically significant increase in the mast cells count in STs in comparison with normal skin. Conclusion Higher expression of MCT in ST compared with normal skin and this enzyme mediator may be responsible for induction of fibrosis in ST.