OVA Group Research Articles

Hyperoxia increases interleukin-17 in airway epithelial cells, alveolar type II cells and alveolar macrophages after ovalbumin-induced lung inflammation

Methods The experimental design was approved by the Ethics Committee of Federal University of Ouro Preto (UFOP)No.092/2012. Groups of female BALB/c mice (8 weeks old; 24.53±0.31g) were divided randomly in to five experimental groups as follows: Control group (CG) remained to air room; PBS group received aluminum hydroxide in phosphate buffered saline (PBS); OVA group (OVA) received ovalbumin and aluminum hydroxide in PBS; O2 group (O2) was exposed to 100% oxygen in a chamber for 24h; OVA+O2 group (OVA+O2) was exposed to 100% oxygen for 24h, received ovalbumin and aluminum hydroxide in PBS. The data were presented as the mean ± standard error of the mean. For continuous data, we used a One-Way Anova followed by the Student–Newman–Keuls post hoc test. For non-continuous data, we used the Kruskal–Wallis test followed by the Dunn’s post hoc test. In all instances, the significance level was set at 5% (p<0.05). Results In bronchoalveolar lavage the hyperoxia decreases macrophage number in O2 (2.82±0.20) and OVA+O2 (1.72±0.15) and increases neutrophils number in O2 (1.79±0.13) and OVA+O2 (1.72±0.15), compared to CG (macrophage: 5.36±0.33) and (neutrophils: 0.02±0.00). The Lymphocytes number were higher in O2 (1.08 ±0.07) and OVA+O2 (0.97±0.08) compared to CG (0.43 ±0.02). When the animals were exposed to oxygen and ovalbumin, concomitantly (OVA+O2 4.55±0.23), the hyperoxia decreases lymphocytes number when compared to OVA (4.55±0.23). The TNF-alpha content were higher in PBS (134.00±7.03), OVA (126.30±3.40), O2 (141.60±6.08) and OVA+O2 (129.60±5.05) when compared to CG (94.67±2.03). In lung sections, the hyperoxia increases interleukin-17 in airway epithelial cells, alveolar type II cells and alveolar macrophages after ovalbumin-induced lung inflammation. When the animals were exposed to oxygen and ovalbumin, concomitantly, the staining with IL17 were increase when compared to OVA (p<0.001).

Chronic low dose chlorine exposure aggravates allergic inflammation and airway hyperresponsiveness and activates inflammasome pathway.

BackgroundEpidemiologic clinical studies suggested that chronic exposure to chlorine products is associated with development of asthma and aggravation of asthmatic symptoms. However, its underlying mechanism was not clearly understood. Studies were undertaken to define the effects and mechanisms of chronic low-dose chlorine exposure in the pathogenesis of airway inflammation and airway hyperresponsiveness (AHR).MethodsSix week-old female BALB/c mice were sensitized and challenged with OVA in the presence and absence of chronic low dose chlorine exposure of naturally vaporized gas of 5% sodium hypochlorite solution. Airway inflammation and AHR were evaluated by bronchoalveolar lavage (BAL) cell recovery and non-invasive phlethysmography, respectively. Real-time qPCR, Western blot assay, and ELISA were used to evaluate the mRNA and protein expressions of cytokines and other inflammatory mediators. Human A549 and murine epithelial (A549 and MLE12) and macrophage (AMJ2-C11) cells were used to define the responses to low dose chlorine exposure in vitro.ResultsChronic low dose chlorine exposure significantly augmented airway inflammation and AHR in OVA-sensitized and challenged mice. The expression of Th2 cytokines IL-4 and IL-5 and proinflammatory cytokine IL-1β and IL-33 were significantly increased in OVA/Cl group compared with OVA group. The chlorine exposure also activates the major molecules associated with inflammasome pathway in the macrophages with increased expression of epithelial alarmins IL-33 and TSLP in vitro.ConclusionChronic low dose exposure of chlorine aggravates allergic Th2 inflammation and AHR potentially through activation of inflammasome danger signaling pathways.

Airway remodeling is reversed by aerobic training in a murine model of chronic asthma.

The aim of this study was to investigate if the aerobic training (AT) reverses airway remodeling (AR) in an asthma model. BALB/c were divided into four groups: control (unsensitized and untrained); ovalbumin (OVA: sensitized and untrained); AT (unsensitized and trained) and OVA + AT. Allergic inflammation was induced with intraperitoneal and OVA inhalation. AT (low intensity; 5×/week; 60 min/session) was performed at 7, 15, and 30 days. Leukocyte counting in the bronchoalveolar lavage fluid; the expression of IL-5, eotaxin, RANTES, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1); AR features (airway smooth muscle, epithelium thickness, collagen and elastic fibers, mucus production); and AR inducers (transforming growing factor-beta, osteopontin, vascular endothelial growth factor). OVA induced an increase in leukocyte airway migration and increased AR features (P < 0.05). After 7 days, AT reversed the OVA-induced eosinophil and macrophage airway migration, the expression of IL-5, eotaxin, RANTES, ICAM-1, VCAM-1, and all AR inducers. However, total reversion of the AR features and inducers and airway inflammation occurred only after 15 days of AT compared with the OVA groups (P < 0.05) and the effects were maintained until the 30th day. AT reverses AR after 15 days and this effect is preceded by the inhibition of leukocyte migration and occurs simultaneously with the reduction in the expression of inflammatory mediators and AR inducers.

Influence of Oral Tolerance on Lung Cytokines Expression and Oxidative Stress Activation in Guinea Pigs with Chronic Inflammation

Objective: We had previously demonstrated that oral induced tolerance attenuates lung tissue hyperresponsiveness, eosinophil inflammation and extracellular matrix remodelling in a model of chronic inflammation in guinea pigs. In the present study, we evaluated if these responses were associated to alterations on Th1/Th2 cell expression on airways and distal lung. Methods: Animals received seven inhalations of ovalbumin (1-5 mg/mL; OVA group) or saline (SAL group) during 4 wk. Oral tolerance (OT) was induced by offering ad libitum ovalbumin 2% in sterile drinking water starting with the 1st inhalation (OT1 group) or after the 4th (OT2 group). After the last inhalation, lungs were removed for the histological analysis using morphometry. We assessed IL-2, IL-4, IL-13, IFN-γ and iNOS both in airways and distal lung. Results: There was an increase in IL-2, IL-4, IL-13, IFN-γ and iNOS positive cells both in airways and alveolar septa in ovalbumin-exposed guinea pigs compared with controls (P<0.05). Both in airways and in lung tissue there was a decrease in IL-4, IL-13 and iNOS positive cells in OT1 and OT2 compared to OVA (P<0.05). Considering IL-2 expression, there was an increase in OT1 and OT2 compared to OVA (P<0.05). We observed positive correlations among the functional responses and some inflammation and oxidative stress pathway activation markers evaluated, especially in alveolar wall. Conclusion: Oral tolerance induces a shift in Th1/Th2 and influences oxidative stress activation both in airways and distal lung of animals with chronic pulmonary allergic inflammation. These results may clarify the mechanisms involved in the attenuation of mechanical responsiveness, inflammation and remodelling of airways and distal lung by oral tolerance, as previously shown in this animal model.

Effects of Rho-kinase inhibition in lung tissue with chronic inflammation

We evaluated whether Rho-kinase inhibition (Y-27632) modulated distal lung responsiveness, inflammation, extracellular matrix remodeling and oxidative stress activation in guinea pigs (GPs) with chronic allergic inflammation. GPs were submitted to inhalation of ovalbumin (OVA-2×/week/4 weeks). From the 5th inhalation on, the Rho-kinase inhibitor group animals were submitted to Y-27632 inhalation 10min before each inhalation of OVA. Seventy-two hours after the seventh inhalation, the oscillatory mechanics of the distal lung strips were assessed under the baseline condition and after the ovalbumin challenge. Subsequently, the lung slices were submitted to morphometry. Rho-kinase inhibition in the ovalbumin-exposed animals attenuated distal lung elastance and resistance, eosinophils, IL-2, IL-4, IL-5, IL-13, TIMP-1, MMP-9, TGF-β, IFN-γ, NF-κB and iNOS-positive cells and the volume fraction of 8-iso-PGF2α, elastic, collagen and actin in alveolar walls compared with the OVA group (P<0.05). Rho-kinase inhibition contributed to the control of distal lung responsiveness, eosinophilic and Th1/Th2 responses and extracellular matrix remodeling in an animal model of chronic allergic inflammation.

Bone marrow-derived mononuclear cells vs. mesenchymal stromal cells in experimental allergic asthma

We compared the effects of bone marrow-derived mononuclear cells (BMMCs) and mesenchymal stromal cells (MSCs) on airway inflammation and remodeling and lung mechanics in experimental allergic asthma. C57BL/6 mice were sensitized and challenged with ovalbumin (OVA group). A control group received saline using the same protocol. Twenty-four hours after the last challenge, groups were further randomized into subgroups to receive saline, BMMCs (2×106) or MSCs (1×105) intratracheally. BMMC and MSC administration decreased cell infiltration, bronchoconstriction index, alveolar collapse, collagen fiber content in the alveolar septa, and interleukin (IL)-4, IL-13, transforming growth factor (TGF)-β and vascular endothelial growth factor (VEGF) levels compared to OVA-SAL. Lung function, alveolar collapse, collagen fiber deposition in alveolar septa, and levels of TGF-β and VEGF improved more after BMMC than MSC therapy. In conclusion, intratracheal BMMC and MSC administration effectively modulated inflammation and fibrogenesis in an experimental model of asthma, but BMMCs was associated with greater benefit in terms of reducing levels of fibrogenesis-related growth factors.

Bone marrow mononuclear cell therapy in experimental allergic asthma: Intratracheal versus intravenous administration

We hypothesized that the route of administration would impact the beneficial effects of bone marrow-derived mononuclear cell (BMDMC) therapy on the remodelling process of asthma. C57BL/6 mice were randomly assigned to two main groups. In the OVA group, mice were sensitized and challenged with ovalbumin, while the control group received saline using the same protocol. Twenty-four hours before the first challenge, control and OVA animals were further randomized into three subgroups to receive saline (SAL), BMDMCs intravenously (2×106), or BMDMCs intratracheally (2×106). The following changes were induced by BMDMC therapy in OVA mice regardless of administration route: reduction in resistive and viscoelastic pressures, static elastance, eosinophil infiltration, collagen fibre content in airways and lung parenchyma; and reduction in the levels of interleukin (IL)-4, IL-13, transforming growth factor-β and vascular endothelial growth factor. In conclusion, BMDMC modulated inflammatory and remodelling processes regardless of administration route in this experimental model of allergic asthma.

NPY and NPY receptors in airway structural and inflammatory cells in allergic asthma

PurposeNeuropeptide Y (NPY) level is elevated in allergic asthmatic airways and activation of NPY receptor-1 (NPY-Y1) on antigen‐presenting cells (APCs) is essential for T cell priming. Paradoxically, NPY-Y1 modulates hyper-responsiveness in T cells, suggesting a bimodal role for NPY in APCs and T cells. Therefore, determination of the temporal and spatial expression pattern of NPY and its receptors in asthmatic airways is essential to further understand the role of NPY in allergic asthma. MethodsLungs were isolated from control and acute and chronic stages of OVA-sensitized and challenged mice (OVA). Stains, including H&E, PAS, and trichrome, were used to determine the severity of lung pathology. The expression patterns of NPY and NPY-Y receptors in the airways were determined using ELISA and immunofluorescence. Cytokine levels in the BALF were also measured. ResultsNPY levels were undetectable in the BALF of control mice, but significantly increased in the OVA group at day 80. Levels of IL-4, TGF-β1 and TGF-β2, significantly increased and peaked on day 45 and decreased on day 80 in the OVA group, exhibiting an inverse correlation with NPY levels. NPY expression was localized to macrophage-like cells in the peri-bronchial and peri-vascular areas in the lung tissue. NPY-Y1 and -Y5 receptors were constitutively expressed by both structural and inflammatory cells in the lung tissue. ConclusionsNPY produced by activated macrophage-like cells may be involved in regulating cytokine production and cellular activities of immune cells in asthma. However, it remains unclear whether such an increase in NPY is a defensive/compensatory mechanism to modulate the effects of inflammatory cytokines.

Haptoglobin baseline value in jennies and the effect of ovariectomy on its serum concentration

The study was conducted to determine the baseline concentration of serum haptoglobin (Hp) in jennies during the breeding and nonbreeding season and to evaluate the effects of ovariectomy on serum Hp concentrations in jennies. Eighteen adult jennies were divided in three groups: nine jennies (OVA) were ovariectomized using laparoscopic surgery, six jennies (LAP) were exploratory examined by laparoscopic surgery, and three jennies were used as a control group. Blood samples were collected from the animals at Day −6, −2, −1, 0, 1, 2, 5, 8, 15, 22, 29 and 36 of surgery. Serum samples were analyzed by an ELISA specifically developed for determining equine Hp. The mean weekly Hp concentration ranged between 149.76±7.55 and 178.94±6.67mg/L. The Hp concentrations of clinically healthy jennies revealed no significant variation among time, and there was no effect of reproductive season on Hp concentrations in jennies. Serum Hp concentration was elevated at the first day after operations in the OVA and LAP group. Five days after the operation, the Hp concentration reached the maximum in the LAP and OVA group (278.84±34.22 and 359.88±35.45mg/L, respectively) and decreased at Day 8 after the operations. On Day 22, 29 and 32 after the operations, the concentration of Hp in LAP and OVA animals was close to its concentration in the control group. In conclusion, Hp is not related to reproductive status of jennies and it can be used as an indicator for cell and tissue damage after surgical operations.

Prolonged allergen challenge in a guinea pig model of allergic rhinitis leads to nasal mucosa remodeling

The aim of this study was to use a guinea pig model of prolonged allergic-induced rhinitis to characterize the feature of nasal mucosa remodeling. Forty-eight male Hartley guinea pigs were randomly divided into six groups: allergen challenged groups (Group OVA(2w) , Group OVA(6w) and Group OVA(12w)) and control groups respectively (Group Sal(2w), Group Sal(6w) and Group Sal(12w)). Each group had 8 guinea pigs. To develop a guinea pig model of nasal mucosa remodeling, ovalbumin-sensitized guinea pigs were repeatedly challenged with allergen twice a week from two weeks to 12 weeks. Matched control groups were challenged with physiological saline. Nasal lavage was performed 24 hours after the last intranasal challenge. Then nasal mucosa were obtained. HE, AB-PAS, MT, and immunohistochemical staining against transforming growth factor-beta1 (TGF-beta1) were performed. Eosinophil cationic protein (ECP) and OVA-special IgE (OVA-sIgE) were detected by ELISA in nasal lavage fluid. (1) The levels of OVA-sIgE in nasal lavage fluid in Group OVA(6w) and Group OVA(12w) were significantly different from Group OVA(2w), while the levels of ECP had no significant difference among the experiment groups. The levels of OVA-sIgE and ECP in experiment groups were significantly different from control groups respectively (P < 0.01). (2) Grade 0 and Grade 1 of epithelial damage were significantly different in Group OVA(6w) and Group OVA(12w) when compared with from Group OVA(2w) (P < 0.01). At the same time, Grade 0 and Grade 1 of epithelial damage were statistically different in the experiment groups when compared with the respectively control groups (P < 0.05). (3) Goblet gland hyperplasia and collagen deposit within the extracellular matrix (ECM) were easily found in Group OVA(6w) and Group OVA(12w) compared with Group OVA(2w) (P < 0.01). The number of goblet gland and the ratio of collagen deposit were statistically more in Group OVA(6w) and Group OVA(12w) than in Group Sal(6w) and Group Sal(12w) (P < 0.05). That feature of the ratio of collagen deposit did not show in Group OVA(2w) versus Group Sal(2w). (4) Increased TGF-beta1 expressions were observed in Group OVA(6w) and Group OVA(12w) compared with Group OVA(2w) (P < 0.01). Those increasing expressions were also observed in experiment groups rather than in the respectively control groups (P < 0.01). Epithelial damage, goblet cells hyperplasia and collagen deposition in ECM were observed as the features of remodeling in this guinea pig model of allergic rhinitis under prolonged allergen challenge. Epithelial damage, excessive expression of related cytokines and enhancement activity of enzymes were observed in early time after challenge of allergen. The features of goblet cells hyperplasia and collagen deposition in ECM were observed at a later stage.

The inhibitory effects and mechanism of Pyrin recombination protein on airway inflamation in mouse with bronchial asthma

Objective To investigate the blocking effects of Pyrin domain protein on mouse allergic airway inflammation induced by ovalbumin and its mechanism.Methods Forty male BALB/c mice were randomly divided into 4 groups with 10 mice in each group.Control group:mice were treated with saline;OVA group:mice were sensitized and challenged with OVA; Pyrin domain protein 3 d group:mice were sensitized and challenged as asthmatic group and treated with 100 μg/kg triptolide before challenged; Pyrin domain protein 7 d group:mice were sensitized and challenged as asthmatic group and treated with 100 μg/kg triptolide before challenged.All mice were killed 24 h after final OVA challenge.The left lung was isolated for pathological examination.Lung sections were stained with hematoxylin and eosin(HE),Masson's trichrome.The thickness of bronchial airway,bronchial smooth muscle thickness,and the collagen deposition area were measured by image analysig system.The concentrations of IL-4,IL-5,TNF-α and IFN-γ in BALF were measured by ELISA,the RT-PCR and Western blot was performed to detect the mRNA expression of connective tissue growth factor (CTGF),TGF-β 1 mRNA and NF -kB from the right lung tissues.Results In OVA group,the number of inflammatory cells and the concentrations of IL-4,IL-5,TNF-a in BALF and Bronchial airway thickness,bronchial smooth muscle thickness,the collagen deposition area,CTGF,TGF-β1 mRNA and NF-kB in lung tissue were significantly higher than those in control group(P＜0.05).IFN-γ were lower than those in control group( P＜0.05 ).While in Pyrin domain protein 3 d group and Pyrin domain protein 7 d group,the number of inflammatory cells,the concentrations of IL-4,IL-5,TNF-α in BALF and bronchial airway thickness,bronchial smooth muscle thickness,the collagen deposition area and NF-kB in lung tissue were significantly lower than those in OVA group,IFN-γwere higher than those in OVA group( P ＜0.05).In Pyrin domain protein 7 d group CTGF and TGF-β1 mRNA in lung tissue were significantly lower than those in OVA group.Conclusion Pyrin domain protein might inhibits airway inflammation and development of airway remodeling in asthmatic mouse,the possible mechanism might in part by associated with reducing the expression of NF-kB and further inhibiting CTGF,TGF-β1. Key words: Bvonchial asthma; Airway remodeling; Mice; Pyrin recombinant protein

Oral Administration Recombinant Bifidobacterium-LTB (B Subunit of Heat-Labile Enterotoxin) Enhances the OVA (Ovalbumin)-Specific sIgA in Jejunal Mucosa of Sprague-Dawley (SD) Rat

The LTB of enterotoxigenic Escherichia coli (ETEC) expressed in Bifidobacterium infantis (BI) has been testified as mucosal adjuvant with co-vaccination BI-CfaB (the major fimbrial subunit) together in vivo in our previous study. In order to investigate the mucosal adjuvant effect of BI-LTB to purified antigens, we oral vaccinated SD rats with recombinant BI-LTB plus OVA (rBI-LTB + OVA), and wild type BI plus OVA (wBI + OVA), OVA and PBS (Phosphate buffered saline) were vaccinated as controls, respectively. The OVA-specific sIgA in jejunal mucosa and specific IgG in serium were measured with ELISA (Enzyme-linked immunosorbent assay) and the sIgA producing cells were detected with immunohistochemistry technology (IHC) and Qwin image manipulation tools subsequently. The results shown rBI-LTB could stimulate SD rats produce high titer OVA-specific sIgA in rBI-LTB + OVA group and the OVA-specific sIgA titer in rBI-LTB + OVA group was found significant greater than that of the wBI + OVA group or OVA single group (p < 0.05). However no such significant difference was detected between the group wBI + OVA and OVA. IHC results suggested that intestinal mucosa and submucosa was the main field of sIgA secretion. These results suggested that recombinant LTB expression in BI could be used as a wide range mucosal adjuvant with different form antigens.

Residual oil fly ash worsens pulmonary hyperreactivity in chronic allergic mice

BALB/c mice received saline (SAL groups) or ovalbumin (OVA groups) intraperitoneally (days 1, 3, 5, 7, 9, 11 and 13). After 27 days, a burst of intratracheal OVA or SAL (days 40, 43 and 46) was performed. Animals were then divided into four groups (N=8, each) and intranasally instilled with saline (SAL-SAL and OVA-SAL) or residual oil fly ash (SAL-ROFA and OVA-ROFA). 24h later, total, initial and difference resistances (Rtot, Rinit, Rdiff) and static elastance (Est) were measured. Lung responsiveness to methacholine was assessed as slope and sensitivity of Est, Rtot, Rinit, and Rdiff. Lung morphometry (collapsed and normal areas and bronchoconstriction index) and cellularity (polymorphonuclear, mononuclear and mast cells) were determined. OVA or ROFA similarly impaired lung mechanics and increased the amount of polymorphonuclear cells and collapsed areas. OVA-ROFA showed even higher hyperresponsiveness, bronchoconstriction and mast cell infiltration. Thus, we concluded that ROFA exposure may add an extra burden to hyperresponsive lungs.

Salbutamol improves markers of epithelial function in mice with chronic allergic pulmonary inflammation

We investigated the effects of salbutamol on the markers of epithelial function in a murine model of chronic allergic pulmonary inflammation by recording the ciliary beat frequency (CBF) and the transepithelial potential difference (PD) in vivo. Mice were sensitized and received four challenges of ovalbumin (OVA group) or 0.9% saline (control group). Forty-eight hours after the 4th inhalation, we observed eosinophilia in the bronchoalveolar lavage and epithelium remodeling with stored acid mucus in the OVA group (P < 0.001). No difference in the baseline CBF was noticed between the groups; however, the OVA group had a significantly lower baseline PD (P = 0.013). Salbutamol increased the CBF in all groups studied, and the dose response curve to salbutamol increased the PD in the OVA group from 10(-4)M to 10(-2)M. We suggest that salbutamol affects the CBF and the depth of the periciliary layer, which, in great part, determines the ability of the cilia to propel the mucus layer. This effect may have a positive impact on airway mucociliary transport in asthma and may have clinical implications.

Intranasal pretreatment with toll like receptor 9 ligand CpG oligodeoxynucleotides prevents the development of allergic rhinitis in juvenile guinea pigs

To evaluate the therapeutic effect of intranasal oligodeoxynucleotides with CpG motifs (CpG ODN) in prevention of allergic rhinitis in juvenile guinea pigs. Juvenile guinea pigs aged from 7 to 10 weeks were administrated with CpG ODN alone or combined with OVA at single dose concentration intranasally (on day 0, 5, 10, 15 in sequence) while control and blank group were administrated with saline. Both experimental and control animals were again sensitized by OVA (on day 18, 25), and 14 days after second sensitization animals were challenged by OVA intranasally (on days 39 and 46). Two hours after challenge, the animals were sacrificed. Then Hemotoxin and Eosin stain were carried out to analyze local eosinophilic reactions and nasal lesions. Local and systemic cytokines interleukin IL-5 and IFN-γ levels were examined by ELISA. Immunofluorescence was carried out with ICAM-1 antibody. Statistical analysis was performed using a SPSS 11.0 software. In CpG ODN-administration or CpG ODN with OVA-administration group allergic rhinitis symptoms were not as severe as model control group (P < 0.05). Compared with the model control group, CpG ODN-administration did not increase production of OVA-specific Th1 cytokine IFN-γ but decreased productions of ovalbumin-specific Th2 cytokines IL-5 both in serum and nasal specimen (q value were 3.890 and 4.019, P < 0.05). Moreover, nasal lesions with infiltration of mean (x ± s) eosinophils (20.0 ± 9.6) in CpG group animal were prominently reduced by the CpG ODN-treatment compared with the control animals (53.5 ± 19.8) and CpG+OVA group (9.5 ± 5.7) were lower than CpG-M+OVA group (49.2 ± 18.9), the differences were significant (q value were 3.785 and 4.576, P < 0.05). Immunofluorescence results showed lower ICAM-1 expression in nasal specimen of CpG group compared with model group and CpG plus OVA group animal to CpG mimics plus OVA group (Z value were 3.697 and 3.765, P < 0.05). Intranasal administration of CpG oligodeoxynucleotides with or without allergen may be an effective way to prevent the development of allergic rhinitis.

TSLP promotes lung Inflammation via activating dendritic cells in OVA-induced mice asthmatic model

Objective To study the expression of thymic stromal lymphopoietin(TSLP) and the activation of DCs in OVA-induced murine asthma model, and investigate the effects and underlying mecha-nisms of TSLP on lung inflammation. Methods Thirty BALB/c mice were randomly divided into control group, OVA group and TSLP neutralizing antibody treated group. The asthma model was evaluated by airway responsiveness and histological analysis of lung tissues ; The levels of TSLP mRNA in lungs were determined by quantitative real-time PCR; The expression of TSLP in lungs were determined by immunohistochemistry and Western blot; The expression of CD40, CD80, CD86 in BALF was detected by FACS. Results Both the histological analysis of lung tissues and the airway responsiveness were all consistent with the characteris-tic of murine asthma model. The expression of TSLP and TSLP mRNA in the OVA group was significantly in-creased compared with blank group. The expression of CD40, CD80, CD86 in BALF from OVA group was increased significantly compared with the control group. Furthermore, treating mice with TSLP neutralizing antibody reduced the expression of CD40, CD80, CD86 on dendritic cells, and IL-4, IL-5, IL-13 in the OVA group. Conclusion Our study indicate that TSLP was highly expressed in the bronchial epithelia of murine asthma model, via upregulation of CD40, CD80, CD86, induce DCs to active CD4~+ T cells and pro-duce type 2 responses, so that aggravating the lung inflammation of asthma. Blocking TSLP is capable of in-hibiting the production of Th2 cytokines, thus presents a promising strategy for the treatment of asthma. Key words: Airway epithelium; TSLP; Dendritic cell; Bronchial asthma

The reproductive biology and the histological and ultrastructural characteristics in ovaries of the female gadidae fish Merluccius merluccius from the Egyptian Mediterranean water

Monthly analysis of the maturity stages distribution revealed that the gadidae fish Merluccius merluccius has a long spawning period extending from early January to early June. All the females over 34.4 cm in body length are mature. The peak value of gonadosomatic index (GSI) was attained in January and continued to May and thereafter decreased gradually from June to August. The analysis of ova diameter revealed that there are eight ova groups in each ripe and spawning ovary; the first four groups are small and transparent and the remaining four groups are yolky. Fecundity shows a wide range for a given length, the absolute fecundity has a linear relationship with length groups and gutted weight. Relative fecundity ranged from 743 to 1699 egg per 1 cm and varied from 92 to 148 eggs per 1 g. The general pattern of the histological development of the ovaries includes six periods: 1. Immaturation period characterized by “small spherical cells each with a large nucleus”. 2. The maturation period characterized by “appearance of isolated follicular epithelial cells around the oocyte and the formation of yolk nuclei”. 3. Vacuolization period characterized by appearance of “marginal vacuoles; the oocyte wall consists of zona radiata coated with a follicular epithelial layer”. 4. The yolk deposition period characterized by “the presence of yolk granules in the cytoplasm”. 5. Ripening period characterized by “migration of the nucleus to the animal pole”. 6. Spawning-spent period characterized by presence of “empty follicles, different stages of cytoplasmic growth”. The ultrastructure of the vacuolized and ripe oocyte wall showing the presence of five different layers, in which the outer most layers, is theca layer; the second layer is the follicular epithelial layer and then the third and fourth layers are zona radiata externa and zona radiata interna, respectively. The fifth layer is known as “cortical alveoli. Key words: Merluccius merluccius, reproductive biology, oogenesis- histology, ultrastructure.

Fouling and boring organisms that deteriorate various European and tropical woods at Turkish seas

Monthly analysis of the maturity stages distribution revealed that the gadidae fish Merluccius merluccius has a long spawning period extending from early January to early June. All the females over 34.4 cm in body length are mature. The peak value of gonadosomatic index (GSI) was attained in January and continued to May and thereafter decreased gradually from June to August. The analysis of ova diameter revealed that there are eight ova groups in each ripe and spawning ovary; the first four groups are small and transparent and the remaining four groups are yolky. Fecundity shows a wide range for a given length, the absolute fecundity has a linear relationship with length groups and gutted weight. Relative fecundity ranged from 743 to 1699 egg per 1 cm and varied from 92 to 148 eggs per 1 g. The general pattern of the histological development of the ovaries includes six periods: 1. Immaturation period characterized by “small spherical cells each with a large nucleus”. 2. The maturation period characterized by “appearance of isolated follicular epithelial cells around the oocyte and the formation of yolk nuclei”. 3. Vacuolization period characterized by appearance of “marginal vacuoles; the oocyte wall consists of zona radiata coated with a follicular epithelial layer”. 4. The yolk deposition period characterized by “the presence of yolk granules in the cytoplasm”. 5. Ripening period characterized by “migration of the nucleus to the animal pole”. 6. Spawning-spent period characterized by presence of “empty follicles, different stages of cytoplasmic growth”. The ultrastructure of the vacuolized and ripe oocyte wall showing the presence of five different layers, in which the outer most layers, is theca layer; the second layer is the follicular epithelial layer and then the third and fourth layers are zona radiata externa and zona radiata interna, respectively. The fifth layer is known as “cortical alveoli.

Effect of inactivated schistosome ova on intestinal epithelial tight junctions during tinitrobenzene sulfonic acid-induced colitis

Objective To observe the possible effect of inactivated schistosome ova on the expression of intestinal epithelial tight junctions ZO-1 and Occludin gene in mouse colitis induced by tinitrobenzene sulfonic acid (TNBS) and its mechanism. Methods Fifty mice were divided into control group (group A, n= 10),TNBS plus normal saline(NS) group(group B, n= 20) and TNBS plus inactivated schistosome ova group(group C, n= 20). Group C was exposed to 10 000 freeze-killed schistosome ova by intraperitoneal injection at day 14 and day 3 before colitis induction. Meanwhile,group B was exposed to 1 ml NS by intraperitoneal injection. The mice in group B and C were challenged with 3 mg TNBS to induce colitis. All mice were killed 7-day after colitis induction and assessed with following variables including mortality, pathological change with HE staining of colon.The transcription levels of ZO-1 and Oceludin in colon tissues were examined using Real-time PCR.The expression and distribution of ZO-1 and Occludin proteins were detected by Western blotting and immunohistochemistry. Results In comparison with group B inactivated schistosome ova most effectively reduced the mortality (30% vs 15 %) and histopathologic severity of TNBS-induced colitis (4.21±0.40 vs 1.74±0.10). The transcription levels of ZO-1 and Occludin in group B were decreased compared with those in group A and group C (P<0.01). When compared with group B,group C showed a significant elevation of the alteration of ZO-1, Occludin proteins expression and localization. Conclusion The results clearly show that schistosome ova treatment reduced the severity of experimental colitis through the regulation of tight junction proteins. Key words: Inflammatory bowel diseases; Schistosorna; Tight junctions protein; Modles, animal

The Soluble Tumor Necrosis Factor-Alpha Receptor Suppresses Airway Inflammation in a Murine Model of Acute Asthma

PurposeTumor necrosis factor-alpha (TNF-α) is a proinflammatory cytokine that has been implicated in many aspects of the airway pathology in asthma. TNF-α blocking strategies are now being tried in asthma patients. This study investigated whether TNF-α blocking therapy inhibits airway inflammation and airway hyperresponsiveness (AHR) in a mouse model of asthma. We also evaluated the effect of TNF-α blocking therapy on cytokine production and adhesion molecule expression.Materials and MethodsOvalbumin (OVA) sensitized BALB/c female mice were exposed to intranasal OVA administration on days 31, 33, 35, and 37. Mice were treated intraperitoneally with soluble TNF-α receptor (sTNFR) during the OVA challenge.ResultsThere were statistically significant decreases in the numbers of total cell and eosinophil in bronchoalveolar lavage fluid (BALF) in the sTNFR treated group compared with the OVA group. However, sTNFR-treatment did not significantly decrease AHR. Anti-inflammatory effect of sTNFR was accompanied with reduction of T helper 2 cytokine levels including interleukin (IL)-4, IL-5 and IL-13 in BALF and vascular cell adhesion molecule 1 expression in lung tissue.ConclusionThese results suggest that sTNFR treatment can suppress the airway inflammation via regulation of Th2 cytokine production and adhesion molecule expression in bronchial asthma.