Effects of hyperbaric oxygen therapy on autistic behaviors and GRIN2B gene expression in valproic acid-exposed rats.

Introduction: Autism is a complex neurodevelopmental condition characterized by deficits in social interaction, communication, and restricted repetitive behaviors. Hyperbaric oxygen therapy (HBOT) has emerged as a potential treatment for autism, although its effects on behavior and gene expression are not well understood. The GRIN2B gene, known for its involvement in encoding a glutamate receptor subunit crucial for neuron communication and associated with autism, was a focus of this study. Methods: Using a rat model induced by prenatal exposure to valproic acid, we examined the impact of HBOT on autism-like behaviors and GRIN2B gene expression. Male Wistar rats were categorized into four groups: control, VPA (valproic acid-exposed), VPA+HBOT [2 atmosphere absolute (ATA)], and VPA+HBOT (2.5 ATA). The rats underwent several behavioral tests to assess social behavior, anxiety, stereotype and exploratory behaviors, and learning. Following the behavioral tests, the HBOT groups received 15 sessions of HBOT at pressures of 2 and 2.5 (ATA), and their behaviors were re-evaluated. Subsequently, real-time PCR was employed to measure GRIN2B gene expression in the frontal lobe. Results: Our results indicated that HBOT significantly increased social interaction and exploratory behaviors in VPA-exposed rats, alongside elevated GRIN2B gene expression in their frontal lobe. Discussion: Our findings imply that HBOT might have a potential role in ameliorating autism-related behaviors in the VPA rat model of autism through potential modulation of GRIN2B gene expression. However, additional research is essential to fully comprehend the underlying mechanisms and refine the HBOT protocol for optimizing its effectiveness in improving autism-related symptoms.

The association between MUC5AC and MUC5B genes expression and remodeling progression in severe neutrophilic asthma: A direct relationship.

BACKGROUND: MUC5 dysregulation is a hallmark of severe neutrophilic asthmatic patients. This study investigates the expression of MUC5AC and MUC5B at mRNA levels on asthma severity and airway wall thickness in severe neutrophilic asthmatic patients. METHOD: In this case-control clinical trial, twenty-five severe neutrophilic asthmatic patients and ten control subjects were enrolled. Subjects underwent ACT, pulmonary functions tests, and fractional exhaled nitric oxide (FENO). Also, induced sputum has been obtained to assess the expression of MUC5AC and MUC5B by the real-time PCR. In addition, the thickness of the airway wall was assessed by high-resolution computed tomography (HRCT), and bioinformatic analysis was implemented to approve the selection of the appropriate genes and for further investigations. RESULT: A significant difference was observed between the asthmatic and control in MUC5AC and MUC5B mRNA expression. Meanwhile, the expression of MUC5AC increased remarkably by asthma severity; also, it is associated with airway wall thickness (WT) (both P-value <0.05). The expression of MUC5B in asthmatic patients was lower than in control. There is no significant correlation between MUC5B mRNA level and WT and asthma severity. Notably, MUC5AC transcription level was correlated to sputum neutrophil percentage, while MUC5B transcription level had a positive correlation with sputum macrophages and a negative one with sputum neutrophils. CONCLUSION: In severe neutrophilic asthma, airway wall thickness increases with MUC5AC mRNA overexpression, which is probably related to asthma severity and the formation of mucus plugs. However, the expression of MUC5B was decreased, resulting in poor mucociliary clearance in the airways. TRIAL REGISTRATION: IR.IAU.MSHD.REC.1400.124.

Effect of azithromycin on bronchial wall thickness in severe persistent asthma: A double-blind placebo-controlled randomized clinical trial.

BACKGROUND: Azithromycin reduced airway remodeling in animal models of asthma. However, its effect on human subjects has not been studied yet. This study aimed to investigate the effect of long-term treatment with azithromycin on airways wall thickness in patients with severe persistent asthma. METHODS: In this randomized, double-blind, placebo-controlled clinical trial, patients with severe persistent asthma received azithromycin (250 mg, BID, three days a week), prednisolone (5 mg, BID), or placebo for eight months in three separate groups in addition to the standard therapy. The improvement in right upper lobe apical segmental bronchus (RB1) wall thickness obtained by high resolution computed tomography was set as the primary outcome. Secondary outcomes included: cough severity, dyspnea severity, asthma control test (ACT) score, asthma exacerbation rate, pulmonary function tests, and fractional exhaled nitric oxide (FENO). RESULTS: Seventy-eight out of ninety randomized subjects completed eight months of treatment with azithromycin (n = 25), prednisolone (n = 27), or placebo (n = 26). Bronchial wall thickness percentage did not change significantly in any of the groups. However, the inner radius and lumen area of azithromycin and prednisolone-treated subjects increased significantly (p < 0.05 for both). Azithromycin also significantly improved the dyspnea severity, ACT score, FENO, and FEV1, FEF25-75, and FEV1/FVC (p < 0.05 for all). Cough severity or asthma exacerbation rate did not change significantly after eight months of treatment with azithromycin. CONCLUSION: Long-term treatment with azithromycin increased lumen radius and lumen area in patients with severe persistent asthma. However, there was no significant change in wall thickness in any of the treatment groups. TRIAL REGISTRATION: IRCT.com (IRCT20091111002695N8).

Is Serum TGF-ß1 and TGF-ß2 levels Correlated to Children with Autism Intensity?

Objective: Transforming growth factor-beta (TGF-ß), a group of multifunctional growth factors, plays an important role in the neuron survival and neurodevelopmental functions. Some studies have evaluated the correlation between TGF-ß1 and TGF-ß2 abnormalities and autism spectrum disorders. In this study, we compared the TGF-ß1 and TGF-ß2 levels between autistic and intellectually normal individuals. Materials & Methods: The study population consisted of 39 autistic and 30 age-matched intellectually normal individuals (control group). Blood samples were taken from all individuals, and all patients were divided into 2 groups (mild-to-moderate and severe) according to the childhood autism rating scale. The cytokines levels were measured by Enzyme Linked Immunosorbent Assay (ELISA). Results: The mean concentration of TGF-ß1 was significantly lower (P < 0.0001) in children with autism compared to the control group (25.3 ± 6.5 versus 35.1 ± 9.4 ng/mL, respectively). Also, the mean concentration of TGF-ß2 in children with autism (32.35± 7.75 ng/ mL) was higher compared to those in the control group (30.47± 4.36 ng/mL); however, this difference did not reach statistical significance (P = 0.21). A positive correlation was observed between TGF-ß1 concentration and autism severity (r = 0.41; P = 0.02), whereas a negative correlation was found between TGF-ß2 concentration and autism severity (r = -0.41; P = 0.02). severity (r = 0.41; P = 0.02), whereas a negative correlation was found between TGF-ß2 concentration and autism severity (r = -0.41; P = 0.02). Conclusion: The results of the present investigation suggest that there is a decrease in the levels of TGF-ß1 in the serum of patients with autism and this cytokine may be effective in the treatment of the pathophysiological aspects of autism.

Unilateral Buccinator Flap for Lengthening of Short Palate.

BACKGROUND: Velopharyngeal insufficiency is one of the most frequent complications after cleft palate repair. PURPOSE: To evaluate the results and complications of unilateral Buccinator flap (BMF) in velopharyngeal insufficiency. MATERIALS AND METHODS: During 4 years the authors performed unilateral BMF in all short palates. Age, sex, demographic data, length of palate, cause of short palate, nasopharyngoscopy and videofluroscopy results, hyper nasality, nasal escape, nasal emission, nasal fluid leak, speech evaluation and results, outcome and complications of the treatment were surveyed before surgery and in 1, 3, 6 months after treatment. RESULTS: The authors had 43 patients, 29 below 8 years old and 14 adults. Velopharyngeal gap was between 10 and 27 mm, mean 21 mm. Buccinator flap were measuring 15 to 19 mm in width and 32 to 56 mm in length. The operation time was 80 to 100 minutes, mean 86 minutes.Nasal emission, nasal escape, and nasal leak were treated in all patients.Hyper nasality was completely improved in all of the patients below 8 years old (29 patients) and in 10 patients of the adults (totally 39 patients, 90.6%). And it was improved significantly in other 4 patients (9.4%). The speech evaluation reported between 70% and 86% improvements.The lengthening of the palate was between 12 and 19 mm, mean 17 mm.The satisfaction of the patients was as 0% poor, 2.3% fair, 72.1% good, and 25.6% excellent. CONCLUSION: Unilateral BMF is reliable, promising, and safe flap for lengthening of short palate and it can lengthen the palate up to 19 mm. The time of surgery is very short compared with other methods. It is an anatomical treatment versus pharyngeal flap which is not an anatomical one. Speech improvement will achieve in 70% to 86% patients.

Optimization of PLGA formulation containing protein or peptide-based antigen: Recent advances.

Protein or peptide-based antigens are the most promising forms to generate custom protective immune responses for clinical applications. Over the last decades, poly(lactic-co-glycolic acid) (PLGA) as a biodegradable polymer has gained more attention for delivery of protein and peptide. Besides many appropriate characteristics, to improve its properties to overcome some obstacles such as release profile and it is important instability of antigen during both encapsulation and storage. Therefore, optimized procedures conditions require to be used to maintain the integrity of protein structure under several stress factors in formulation process. In this review article, the properties of PLGA particles, their preparation techniques and strategies for improvement of protein stability during encapsulation into PLGA, release from particle and storage as well as stabilization approaches were summarized. © 2018 Wiley Periodicals, Inc. J. Biomed. Mater. Res. Part A: 106A: 2540-2551, 2018.

Comparison of the Effect of Dexamethasone and Tranexamic Acid, Separately or in Combination on Post-Rhinoplasty Edema and Ecchymosis.

BACKGROUND: Dexamethasone and tranexamic acid are used to decrease post-rhinoplasty periorbital edema and ecchymosis. We compared the impact of each medication separately or in combination in this regard. METHODS: A prospective, randomized triple-blinded study was undertaken on 60 patients who underwent primary open rhinoplasty. They were divided into four groups: Group D (n = 15) received 8 mg dexamethasone, group T (n = 15) received 10 mg/kg tranexamic acid, group DT (n = 15) received both 8 mg dexamethasone and 10 mg/kg tranexamic acid, and group P (n = 15) received neither medication and served as the placebo control group. The medications were given intravenously (IV) 1 h before and three doses every 8 h postoperatively. Digital photographs were taken on the first, third and seventh postoperative days. One expert examiner blinded to the study evaluated the periorbital edema and ecchymosis on a scale of 0-4. Periorbital edema and ecchymosis were examined in all groups. RESULTS: In group D, group T and group DT, periorbital edema and ecchymosis ratings were significantly lower compared with the control group (p < 0.01). No statistically significant difference was seen in preventing or decreasing both periorbital edema and ecchymosis among group D, group T and group DT. CONCLUSION: Tranexamic acid and dexamethasone, separately or in combination, had similar effects in reducing periorbital edema and ecchymosis in open rhinoplasty. Combined application did not show a significantly higher beneficial effect in this regard. LEVEL OF EVIDENCE III: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Artemisinin therapeutic efficacy in the experimental model of multiple sclerosis.

CONTEXT: The immune system through T-helper 1 (Th1) and Th17 cells play a critical role in the pathogenesis of experimental autoimmune encephalomyelitis (EAE), whereas the Th2 responses inhibit myelin degeneration. Artemisinin, as an anti-malaria as its agent, has been used widely in the treatment of malaria, shifts the lymphocyte responses from Th1 to Th2. OBJECTIVE: In this study, we have investigated the therapeutic effects of artemisinin on the EAE treatment. MATERIALS AND METHODS: EAE was induced in the inbred C57BL6 mice. High and low doses of prednisolone and artemisinin were injected daily with the control and test groups, respectively. The spleen and the brain of the mice were removed and used for ELISA and histological studies. RESULTS: The mean weight of mice was significantly (p value < .05) higher in artemisinin-treated group compared with the untreated group, whereas, the mean EAE score of mice was significantly (p value < .05) lower in the artemisinin-treated group compared with the untreated group. The brain histology shows the absence of plaque formation in the artemisinin treated group. The concentration of IFN-γ in the low dose of artemisinin treated group showed significantly (p value < .05) lower in comparison to the untreated group. IL-4 concentration was significantly (p value < .05) higher in the treated groups than the control group. CONCLUSIONS: Since, artemisinin can shift the immune responses from Th1 to Th2, therefore, it can be helpful in the treatment of MS after more investigation.

Evaluation of transforming growth factor-beta1 gene expression in pterygium tissue of atopic patients.

BACKGROUND: The exact pathogenesis of pterygium is still not fully understood. Growth factors are considered to play an important role in the formation of pterygium. Transforming growth factor (TGF)-ß1 is considered to be one of the main mediators of fibroblast stimulation and tissue remodeling in allergic conditions. The objective of the present study was to investigate the association between TGF-ß1 gene expression and pterygium in atopic and nonatopic participants. METHODS: We used questionnaires to record demographic and clinical information from patients who underwent pterygium excision surgery. Skin prick examination was done to confirm or rule out atopy in 30 patients with atopy (Case Group) and 30 individuals without atopy (Control Group). Additionally, measurement of serum immunoglobulin E, cytokines, including interleukin-4 and interferon-γ, and peripheral blood eosinophil count was performed to confirm atopy in 30 consecutive patients (Case Group). A semiquantitative reverse transcription polymerase chain reaction was performed to determine TGF-ß1 gene expression in all individuals. RESULTS: TGF-ß1 mRNA gene expression was significantly higher (p = 0.0001) in atopic patients 2.50 ± 1.11 compared to nonatopic individuals 1.40 ± 0.46. Eosinophil count and serum immunoglobulin E were significantly higher (p = 0.031 and p = 0.001, respectively) in atopic patients compared to the Control Group. Serum interleukin-4 was also significantly higher (p = 0.01) in atopic patients compared with nonatopic individuals. CONCLUSION: Excess expression of TGF-ß1 gene in pterygium tissue of atopic individuals suggests that growth factors play a role in the pathogenesis of pterygium.

Influence of splenectomy on immunoglobulins and complement components in major thalassemia.

BACKGROUND: Thalassemia is the most common anemia with hereditary base in Iran. The aim of this study was to evaluate the humoral immune system and assess the effect of splenectomy on the serum level of immunoglobulins IgG, IgM, and IgG and complement components in patients with major thalassemia. METHODS: This quasi-experimental study (before-after) was performed on 40 patients with major thalassemia that referred to the treatment centers of Babol for splenectomy from March 2011 to March 2013. RESULTS: The mean age of patients under study was 25.38±6.89 years. The results of this study showed that the rate of IgA and IgM had a significant decrease after splenectomy. However, the increase of serum level in IgG in this study was not significant. The serum levels of C3 and CH50 were evaluated in all patients that its reduction was statistically significant, but the decrease of serum levels in C4 was not significant. CONCLUSION: The spleen plays a role in releasing immunoglobulins and starter proteins of complement activation pathways and splenectomy causes reduction in the serum levels of immunoglobulins and complement components.

Measurement of Serum Superoxide Dismutase and Its Relevance to Disease Intensity Autistic Children.

BACKGROUND: Autism is a pervasive disorder and its prevalence increased in recent surveys. An estimated 1 out of every 88 children is affected by autism. Autism disorder symptoms appear before the age of three. It is believed that serum levels of superoxide dismutase may play a role in etiology of autism. MATERIALS AND METHODS: Between October and November 2014, 27 Iranian children from Mashhad city were selected in this study. Given these assumptions, the amount of SOD serum in autistic patients and healthy individuals and correlation between the amount of SOD and autism severity were examined. Blood samples of 30 autistic children and 18 age-matched healthy children were collected between 9 to 11 am. Serum level of SOD in both groups was measured by ELISA method. RESULTS: The mean SOD level in the treatment group (1.04 ±1.33 ng/ml) was significantly lower than the control group (p = 0.001). However, SOD level was not significantly associated with the autism severity (p = 0.667). Conclusions: Decreased serum levels of superoxide dismutase in the early diagnosis of autistic children can be considered as a diagnostic biomarker.

Transforming growth factor beta 1 869T/C and 915G/C polymorphisms and risk of autism spectrum disorders.

BACKGROUND: Transforming growth factor-ß1 (TGF-ß1) has been found to play a crucial role in early central nervous system development. Several studies have illustrated decreased TGF-ß1 levels in sera and brains of autistic children. Two point mutations in the TGF-ß1 signal peptide at 869T/C and 915G/C have been reported to influence TGF-ß1 expression. The aim of the present study was to investigate the correlation of TGF-ß1 polymorphisms and their haplotypes with autism. METHODS: This study was performed on 39 autistic patients and 35 age- and sex-matched normal controls in an Iranian population, using the sequence specific primed-polymerase chain reaction (PCR-SSP) technique. Patients were divided into mild-to-moderate and severe groups according to the childhood autism rating scale. RESULTS: No significant differences were observed for allele, genotype, or haplotype frequencies between the autistics and controls. Only a slight difference was observed in GC25 between the controls and all children with autism. CONCLUSION: Thus, these results indicate that the polymorphisms in TGF-ß1 gene may not play an important role in the development of autism.

The expression of vascular endothelial growth factor in pterygium tissue of atopic patients.

The exact pathogenesis of pterygium has not been completely elucidated. Growth factors have been considered to play a role in pterygium formation. Vascular endothelial growth factor (VEGF) is one of the principal mediators of angiogenesis, fibroblast stimulation and tissue remodeling in allergic conditions. The aim of this study was to compare the association between pterygium and VEGF gene expression between atopic and non-atopic individuals. At first visit, all patients with pterygium underwent blood tests, serum immunoglobulin E (IgE), serum cytokines including interleukin-4 (IL-4) and interferon-γ (IFN-γ) and peripheral blood eosinophil count. After obtaining informed consents, questionnaires were used to obtain demographic and clinical data from patients who underwent pterygium excision surgery. Skin prick test was performed to confirm or rule out atopy in 30 patients with (case group) and 30 patients without (control group) atopy. Pterygium tissues were then removed by surgery. A semi-quantitative reverse transcriptase polymerase chain reaction was performed to determine VEGF gene expression in all patients. Our results illustrated that VEGF mRNA expression in atopic patients was significantly higher than in the non-atopic group (P = 0.01). Eosinophil count, serum IgE and IL-4 were also significantly higher in atopic patients than in the non-atopic group (P = 0.03, 0.001 and 0.001, respectively). However, no significant difference was noted in serum IFN-γ between the two groups (P = 0.06). The excessive expression of VEGF gene in pterygium tissue of patients with atopy suggests that growth factors may play a role in the pathogenesis of pterygium or accelerate its formation.

The preventive effects of natural adjuvants, G2 and G2F on tracheal responsiveness and serum IL-4 and IFN-γ (th1/th2 balance) in sensitized guinea pigs.

OBJECTIVE: The effects of natural adjuvants on lung inflammation and tracheal responsiveness were examined in sensitized guinea pigs. METHODS: The responses of guinea pig tracheal chains and the serum levels of interleukin-4 and interferon-gamma were examined in control pigs and three other groups of guinea pigs: the sensitized group and two other sensitized groups treated with either adjuvant G2 or adjuvant G2F (n=7 for each group). Sensitization of the animals was achieved by injection and inhalation of ovalbumin. RESULTS: The results showed that sensitized animals had increased tracheal responsiveness and increased serum levels of interleukin-4 and interferon-gamma compared to controls (p<0.05 to p<0.001). Treatments with either G2 or G2F prevented the increase in tracheal responsiveness and serum interleukin-4 (p<0.01 to p<0.001). However, the serum levels of interferon-gamma and the interleukin-4-to-interferon-gamma ratio was increased in the treated groups (p<0.001 for all cases). CONCLUSIONS: These results indicate important preventive effects of two natural adjuvants, particularly G2, on the changes in tracheal responsiveness, serum cytokines and the interleukin-4-to-interferon-gamma ratio (T helper 1/T helper 2 balance) in sensitized guinea pigs.

The preventive effects of natural adjuvants, G2 and G2F on tracheal responsiveness and serum IL-4 and IFN-? (th1/th2 balance) in sensitized guinea pigs

OBJECTIVE: The effects of natural adjuvants on lung inflammation and tracheal responsiveness were examined in sensitized guinea pigs. METHODS: The responses of guinea pig tracheal chains and the serum levels of interleukin-4 and interferon-gamma were examined in control pigs and three other groups of guinea pigs: the sensitized group and two other sensitized groups treated with either adjuvant G2 or adjuvant G2F (n = 7 for each group). Sensitization of the animals was achieved by injection and inhalation of ovalbumin. RESULTS: The results showed that sensitized animals had increased tracheal responsiveness and increased serum levels of interleukin-4 and interferon-gamma compared to controls (p<0.05 to p<0.001). Treatments with either G2 or G2F prevented the increase in tracheal responsiveness and serum interleukin-4 (p<0.01 to p<0.001). However, the serum levels of interferon-gamma and the interleukin-4-to-interferon-gamma ratio was increased in the treated groups (p<0.001 for all cases). CONCLUSIONS: These results indicate important preventive effects of two natural adjuvants, particularly G2, on the changes in tracheal responsiveness, serum cytokines and the interleukin-4-to-interferon-gamma ratio (T helper 1/T helper 2 balance) in sensitized guinea pigs. .

Value of the polymerase chain reaction method for detecting tuberculosis in the bronchial tissue involved by anthracosis.

BACKGROUND: Anthracofibrosis is the black discoloration of the bronchial mucosa with deformity and obstruction. Association of this disease with tuberculosis (TB) was approved. The objective of this study was to find the additional benefit of assessment of TB by the polymerase chain reaction (PCR) method. METHODS: Bronchoscopy was performed on 103 subjects (54 anthracofibrosis and 49 control subjects) who required bronchoscopy for their pulmonary problems. According to bronchoscopic findings, participants were classified to anthracofibrosis and nonanthracotic groups. They were examined for TB with traditional methods such as direct smear (Ziehl-Neelsen staining), Löwenstein-Jensen culture, and histopathology and the new method "PCR" for Mycobacterium tuberculosis genome (IS6110). RESULTS: Age, sex, smoking, and clinical findings were not significantly different in the TB and the non-TB groups. Acid-fast bacilli could be detected by a direct smear in 12 (25%) of the anthracofibrosis subjects, and adding the results of culture and histopathology traditional tests indicated TB in 27 (31%) of the cases. Mycobacterium tuberculosis was diagnosed by PCR in 18 (33%) patients, but the difference was not significant. Detection of acid-fast bacilli in control nonanthracosis subjects was significantly lower (3, 6%), but PCR (20, 40%) and accumulation of results from all traditional methods (22, 44%) showed a nonsignificant difference. CONCLUSIONS: The PCR method showed a result equal to traditional methods including accumulation of smear, culture, and histopathology.

TLR4 and TLR2 expression in biopsy specimens from antral and corporal stomach zones in Helicobacter pylori infections.

BACKGROUND: It is not yet known which types of Toll-like receptors (TLRs) are most effective in Helicobacter pylori (H. pylori) recognition. It is also not known which gastric zones have the most prominent roles in TLR-mediated bacterial recognition. The aim of this work was to analyze the expression of TLR2 and TLR4 in biopsy specimens from H. pylori-infected patients. METHODS: Thirty-eight patients with gastrointestinal disorders were divided into four groups in this study. The groups were: (A) H. pylori infection and peptic ulcer (n=15), (B) peptic ulcer only (n=5), (C) H. pylori infection only (n=10) and (D) control, with neither H. pylori infection nor peptic ulcer (n=8). Biopsy specimens from sites of redness or atrophic mucosa from gastric antrum and body in patients with gastritis were collected. RNAs from the antrum and body specimens were isolated. TLR2 and TLR4 mRNA expression was assessed by RT-PCR and quantified as densitometric ratios of TLR2 and TLR4/ß-actin mRNA. RESULTS: In the antral zones of H. pylori-infected patients (Groups A and C) TLR2 and TLR4 expression was significantly greater than in uninfected patients (Groups B and D) regardless of peptic ulcers (p < 0.05). In the gastric body samples TLR2 expression was significantly greater in Group C (H. pylori infection only) than in Group B (peptic ulcer only) and TLR4 expression was significantly greater in group A (H. pylori infection and peptic ulcer) than in Group B (peptic ulcer only) (p < 0.05). No significant differences in expression of TLR4 and TLR2 were observed between samples from the antrum and body in same groups. CONCLUSIONS: We conclude that H. pylori infection leads to significant increase in TLR2 and TLR4 molecules expression in antral region related to the control group. Considering the stimulatory effect of H. pylori on TLRs expression in the gastric tissue, we assume that colonization of H. pylori infection might occurs more in the gastric antral region than in the gastric body.

The effect of natural adjuvants (G2, G2F) on lung inflammation of sensitized guinea pigs.

OBJECTIVE: The effects of natural adjuvants were examined on total and differential WBC counts in lung lavage of sensitized guinea pigs. MATERIALS AND METHODS: In three sensitized groups of guinea pigs including: untreated sensitized animals (S), sensitized animals treated with adjuvant G2 (S+G2) and G2F (S+G2F) as well as non-sensitized group (C) (n=6 for each group), total and differential WBC counts of lung lavage were examined. Sensitization of animals was achieved by injection and inhalation of ovalbumin (OA). RESULTS: The results showed increased total WBC, eosinophil, neutrophil, and basophil counts, and decreased lymphocytes in lung lavage of sensitized animals compared with the control group (p<0.01 for all cases). However, neutrophil, lymphocyte, eosinophil, and basophil counts in lung lavage were decreased in treated groups with either G2 or G2F but total WBC was decreased in lung lavage of treated group with only G2. CONCLUSION: These results indicate important preventive effects of two natural adjuvants, especially G2, on lung inflammation of sensitized guinea pigs.

Effect of verapamil on bronchial goblet cells of asthma: an experimental study on sensitized animals.

INTRODUCTION: Goblet cell hyperplasia (GCH) and mucus hypersecretion in the airway is recognized as an important contributor to morbidity and mortality in asthma and COPD. Verapamil is a calcium channel blocker that binds to the alpha-subunit of L-type calcium channels and inhibits the mucin gene via the calmodulin and CaM kinase pathway. The objective of this study was to determine the in vivo effect of verapamil on GCH and eosinophilic inflammation in sensitized mice. METHODS: Male BALB/c mice were sensitized to ovalbumin using the standard method. Two groups of animals were received verapamil via an intramuscular injection: 1-low dose (0.5 mg/kg/day for two weeks), 2-high dose (1.5 mg/kg/day for two weeks). Serum and bronchoalveolar lavage fluid (BALF) was collected and analyzed for inflammatory cells, interferon-γ and IL-4. The left lung was sent for histopathological evaluation, especially for periodic acid-Schiff (PAS), to identify goblet cells in the epithelium. The degree of inflammatory cell infiltration, including eosinophils, mucus plugging, and smooth muscle thickness of the airways were classified on a semi quantitative scale. RESULTS: Inflammatory cell infiltration in peribronchial and perivascular areas was observed in all sensitized groups. Eosinophils percentage in the BALF significantly decreased in verapamil-treated mice compared with sensitized mice (from 19.8% in asthmatic to 5.4% for low dose and 4.4% for high dose). The ratio of airway goblet cells per epithelial cells were significantly lower in verapamil-treated mice versus sensitized mice (1.57±1.30% for low dose; 1.50±0.93% for high dose versus 12.93±7.55%, P<0.05, respectively). Mucus production of goblet cells decreased significantly in verapamil-treated mice versus sensitized mice (mean score was 1.45±0.30 for low dose; 0.81±1.00 for high dose versus 2.85±0.86 in the sensitized control group, P<0.05, respectively). The concentration of serum and BALF-IFN-γ in verapamil-treated mice markedly increased by the verapamil treatment when compared to sensitized mice (15.1±0.43 versus 4.7±0.96, P<0.05 and 91.8±47.7 versus 14.8±4.6, P<0.01, respectively). CONCLUSION: Verapamil is a useful drug with therapeutic targeting on GCH and a potential way to limit mucous production and improve bronchial inflammation.

Decreased levels of soluble Toll-like Receptor 2 in patients with asthma.

BACKGROUND: Recently, reports have indicated a role for the membrane form of Toll-like Receptor 2 (TLR2) in asthma pathogenesis. In this study we examined soluble TLR2 levels in serum and sputum of asthmatic and healthy subjects. METHODS: Serum and sputum samples were obtained from 33 asthmatic and 19 healthy subjects. The asthmatics were classified into four groups according to the Global Initiative for Asthma. A sandwich ELISA was developed to measure soluble TLR2 (sTLR2) in serum and sputum. TLR2 mRNA expression was determined by semi-quantitative RT-PCR of all sputum samples. RESULTS: The mean sTLR2 levels from serum and sputum of asthmatics were significantly lower than those from healthy subjects. Moreover, sTLR2 concentration decreased concomitantly with asthma severity. The differences observed, however, were not statistically significant. TLR2/GAPDH mRNA of sputum leukocytes was also significantly lower in asthmatics than in healthy subjects. CONCLUSION: This study demonstrated for the first time thatsTLR2 levels are lower in serum and sputum samples from asthmatic than from healthy subjects, and this could be an indicator of TLR2 expression. We also found that sTLR2 concentration in serum decreased concomitantly with an increase of asthma severity clinical score .