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The purpose of this research was to investigate the gene expression levels of inflammatory cytokines interferon (IFN)

Multiple sclerosis (MS) is a chronic autoimmune disorder of central nervous system which is increasing worldwide. Although immunosuppressive agents are used for the treatment of MS disease, nevertheless the lack of non-toxic and efficient therapeutic method is perceptible. Hence, this study aims to evaluate the effect of Contactin-associated protein (Caspr) antibody-, poly ethylene glycol (PEG)- and exosome combined gold nanoparticles (GNPs) in comparison to Glatiramer acetate as a selective treatment of MS disease in the experimental autoimmune encephalomyelitis (EAE) mouse model. EAE was induced in female C57BL/6 mice and 25-day treatment with anti-Caspr-, PEG- and exosome combined GNPs was evaluated. Histopathological examination of spinal cord, regulatory T cells as well as inflammatory pathway including IFN-ɣ and IL-17 and mir-326 were investigated. The results showed the severity of MS symptoms was significantly decreased in all treated groups. Histological examination of the spinal cord indicated the reduced demyelination and immune cell infiltration. Besides, regulatory T cells were significantly increased following all treatments. Remarkably, the cytokine levels of IFN-ɣ and IL-17 as well as mir-326 is altered in treated groups. Taken together, the obtained findings demonstrate that the administration of anti-Caspr-, PEG- and exosome combined GNPs can be considered a potential treatment in MS disease.

Background Pollen is one of the most common allergens that cause respiratory allergies worldwide. Pollen grains from poplars have been reported as important sources of pollinosis in many countries. Objective The aim of the present study was to determine the molecular and immunochemical characterization of Pop n 2, a novel allergen of Populus nigra (P nigra) pollen extract. Methods In this study, the pollen extract of P nigra was analysed by SDS-PAGE, and the allergenic profile was determined by IgE immunoblotting and specific ELISA using the sera of twenty allergic patients. The coding sequence of Pop n 2 was cloned and expressed in the Escherichia coli BL21 (DE3) using plasmid the pET-21b (+). Finally, the expressed recombinant Pop n 2 was purified by affinity chromatography. Results Pop n 2 belongs to the profilin family with a molecular weight of approximately 14 kDa. Pop n 2 is the most IgE-reactive protein (about 65%) in the P nigra pollen extract. The cDNA sequencing results indicated an open reading frame 396 bp that encodes 131 amino acid residues. The results of ELISA and Immunoblotting assays showed that recombinant Pop n 2 could react with the IgE antibody in patients' sera, like its natural counterpart. Conclusion Our data revealed that Pop n 2 is a significant allergen in the P nigra pollen extract. Moreover, we observed that the recombinant Pop n 2 produced by the pET-21b (+) vector in the E colisystem acts as its natural counterpart.

Long-term exposure to airborne particles of 10 µm and less in size (PM10) in dust can lead to increased risk of diseases such as respiratory, cardiovascular, lung cancer and atherosclerosis. The aim of the study was to evaluate the effects of water-soluble PM10 particles in the Middle East Dust (MED) storm in Ahvaz, Iran, on the production of TNF-α by human monocytes. In addition, we assessed the level of induction of apoptosis in isolated A549 human pulmonary epithelial cells. For this purpose, isolated human blood monocytes (250,000 to 300,000 cell/ ml) as well as isolated human pulmonary A549 epithelial cells (100,0000 cell/ ml) were exposed to various concentrations (62.5, 125, 250, 500 µg/ml) of water-soluble PM10 particles for different incubation periods (12, 24, 48 h). The results showed that exposure to PM10 particles increased the production of TNF-α in human monocytes and promoted apoptosis induction in A549 cells, in both concentration and incubation of period-dependent manner. In conclusion, airborne dust particles in Ahvaz city contain active compounds capable of increasing production of the pro-inflammatory cytokine, TNF-α, and inducing apoptosis of lung epithelial cells.

Background Nutritional and environmental benefits of mycoprotein verify its beneficial role on the health of humankind in the next decades. Agro-industrial wastes can be used as cheap substrates to decrease the total cost of product. However, fungi may produce toxins or lead to allergic reactions in consumers. Therefore, the study of the safety and nutritional aspects of this product are very important. Results Fusarium venenatum IR372C was cultured on date wastes and ammonium salts in submerge fermentation. The safety and nutritional issues of produced mycoprotein were investigated including allergy tests and analyses of toxins, as well as existence of toxin genes, and content of heavy metals, metals, amino acids and fatty acids. The results showed that fumonisin genes in F. venenatum IR372C remain without any gene expression during 1 week fermentation. Zearalenone and deoxynivalenol cannot be detected in the fermentation medium after 3 weeks. Prick tests on 30 volunteers demonstrated no sensitivities to mycoprotein. The content of lead was 658 μg kg-1 as the highest heavy metal followed by arsenic, cadmium and mercury at 161, 30.57 and 0 μg kg-1 , respectively. Produced mycoprotein includes essential amino acids at appropriate contents and the ratio of unsaturated to saturated fatty acid was nearly 2:1. Also, calcium, iron, magnesium and zinc were found in mycoprotein, which could have health beneficial impacts on consumers. Conclusion This study has provided information on safety aspects of mycoprotein production by F. venentaum IR372C from date wastes. However, further studies with focus on long-term clinical benefits of diets containing mycoprotein are necessary. © 2020 Society of Chemical Industry.

Fraxinus excelsior (Ash) is a common tree and is important cause of winter–spring pollinosis in many temperate regions in the world. In this study, a high molecular weight allergen from ash pollen was identified. In all, 20 individuals allergic to ash participated in the study. Characterization and immunoreactivity of ash pollen proteins was performed using sodium dodecyl sulfate polyacrylamide electrophoresis (SDS-PAGE), two-dimensional (2D) gel electrophoresis and immunoblotting. Immunoglobulin E (IgE)-binding proteins with apparent molecular mass ranging from 9 to 110 kDa were detected in ash pollen extract. Serum IgE of 7 (35%) patients reacted with a 53-kDa protein band. Analysis of 2D immunoblots showed several IgE-binding proteins. Moreover, mass spectrometry analysis indicated that the 53-kDa allergen was homologous to calreticulin. We defined a novel allergen from F. excelsior pollen with a molecular weight of about 53 kDa. This allergen could be considered as an important high molecular weight allergenic protein for further studies on cross-reactivity and development of diagnostic and therapeutic approaches.

After the early rainfall in the autumn of 2013, respiratory syndromes spread in the Khuzestan province of Iran with the most severity in Ahvaz. There have been recurring outbreaks in recent years. Considering that pollen-derived airborne allergens are regarded as key aeroallergens and the main cause of allergic rhinitis and asthma, this work aimed to forecast total pollen concentration in Ahvaz through an artificial neural network (ANN), followed by evaluating the pollen spatial distribution across the city and the association between pollen concentrations and environmental parameters. The utilized ANN in this work included an input layer with 13 parameters, a hidden layer of five neurons, and an output layer. Data were classified into training, validation, and testing sets. The ANN was implemented with 70% and 80% of data for training. The value of the correlation coefficient for the data validation of these two networks was 0.89 and 0.92, respectively. The results also indicated that despite the difference in the mean concentration of the pollens in various areas of Ahvaz, this difference was not statistically significant (P > 0.05). Furthermore, there was a negative correlation between the concentration of total pollen and relative humidity, precipitation, and air pressure. However, it had a positive correlation with temperature. Consequently, considering the logistical challenges of monitoring bioaerosols in the air, the ANN approach could predict total pollen concentrations. Therefore, in addition to measurements, the ANN technique can be a good tool to enable authorities to mitigate the impact of airborne pollen on people.

Background Systemic sclerosis (SSc) or scleroderma is a multiorgan rheumatoid disease characterized by skin tightening or organ dysfunction due to fibrosis, vascular damage, and autoimmunity. No specific cause has been discovered for this illness, and hence no effective treatment exists for it. On the other hand, due to the lack of diagnostic biomarkers capable of effectively and specifically differentiating the patients, early diagnosis has not been possible. Due to their potent regulatory roles in molecular pathways, microRNAs are among the novel candidates for the diagnosis and treatment of diseases like SSc. MiR-27a is a microRNA known for its role in the pathogenesis of fibrosis and cancer, both of which employ similar signaling pathways; hence we hypothesized that Mir-27a could be dysregulated in the blood of individuals affected by SSc and it might be useful in the diagnosis or treatment of this disease. Methods Blood was collected from 60 SSc patients (30 limited and 30 diffused) diagnosed by rheumatologist according to ACR/AULAR criteria; following RNA isolation and cDNA synthesis; real-time qPCR was performed on the samples using Taq-Man probes and data were analyzed by the ΔΔCT method. Also, potential targets of miR-27a were evaluated using bioinformatics. Results It was revealed that miR-27a was significantly down-regulated in SSc patients in comparison to the healthy individuals, but there was no difference in miR-27 expression between limited and diffused SSc patients. Besides, miR-27a was found to target several contributing factors to SSc. Conclusion It seems that miR-27a has a protective role in SSc, and its downregulation could result in the disease's onset. Based on bioinformatics analyses, it is speculated that miR-27a likely targets factors contributing to the pathogenesis of SSc, which are elevated upon the downregulation of miR-27a; hence, miR-27a mimics could be considered as potential therapeutic agents for the treatment of SSc in future studies. Since no difference was observed between limited and diffused patient groups, it is unlikely that this microRNA has a role in disease progression. According to ROC analysis of qPCR data, miR-27a could be employed as a valuable diagnostic biomarker for SSc.

Seyed Ahmad Hosseini,1 Nazanin Zahrooni,2 Ahmad Ahmadzadeh,3 Kambiz Ahmadiangali,4 Mohammad-Ali Assarehzadegan5 1Nutrition and Metabolic Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 2Department of Nutrition, Faculty of Paramedicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 3Thalassemia and Hemoglobinopathy Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 4Biostatistics Division, Health School, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 5Immunology Research Center, Institute of Immunology and Infectious Diseases, Iran University of Medical Science, Tehran, IranCorrespondence: Nazanin ZahrooniDepartment of Nutrition, Faculty of Paramedicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Tel +98 61 3336 7543Fax +98-6133720299Email nzahroonii@gmail.comBackground: Survival rates among breast cancer patients and the number of patients living with treatment side effects have improved, leading to increased focus on quality of life (QOL). The objective of this study was to determine the efficacy of CoQ10 on QOL scores among breast cancer patients in Iranian undergoing tamoxifen therapy.Methods: Thirty breast cancer patients were randomized into two groups. The first group received 100 mg CoQ10, and the second group took fplacebo once a day for 8 weeks. QOL was evaluated by a standard QOL questionnaire and a specific questionnaire on QOL of breast cancer patients at baseline and the end of the study. Also, physical activity of patients was assessed with the IPAQ questionnaire and dietary intake determined by a 3-day dietary record.Results: The data of 30 subjects were analyzed. According to QOL C30 data, CoQ10 led to a significant increase in physical functioning (P=0.029), emotional functioning (P=0.031), and cognitive functioning (P=0.023) compared to placebo. Symptom scales revealed a notable reduction in appetite loss in the first group (P=0.01). Global health status showed no significant changes in either study arm. On the QOL BR23, progress in functions and decline in symptoms were not statistically significant. Arm symptoms showed significant reduction (P=0.022) in patients that received placebo.Conclusion: This trial indicates that CoQ10 supplementation has effects in ameliorating some dimensions of QOL in breast cancer patients. To generalize the results, larger and longer intervention studies are needed.Clinical Trial Registration: IRCT2015042021874N1.Keywords: breast cancer, CoQ10, quality of life, tamoxifen

Airway epithelial cells secrete Interleukin-33 in response to the different allergens. Several single nucleotide polymorphisms (SNP) of this cytokine have been reported to be involved in the development of asthma. We conducted this study to evaluate the impact of the two most common SNPs of the IL-33 gene (rs1342326 and rs3939286) and environmental factors on the susceptibility to asthma in the Iranian population. In this study, we enrolled 126 asthmatics patients and 300 age, sex-matched controls. Genotyping was performed by real-time PCR using the TaqMan SNP genotyping assay. Moreover, total serum IgE level, eosinophil count, and skin prick test were accomplished and complete history was taken from all the participants. The frequencies of mutant genotypes in both SNPs were significantly higher in asthmatics than controls. C/C genotype of rs1342326 [OR (95% CI) 2.50 (1.33–4.69)] and A/A genotype of rs3939286 [OR (95% CI) 2.18 (1.05–4.52)] were associated with higher risk of asthma development. While A/C+C/C genotype of rs1342326 was more prevalent in mild asthma [OR (95% CI) 2.36 (1.14–4.89)], G/A+A/A genotype of rs3939286 was associated with increased risk of moderate and severe asthma [OR (95% CI) 2.53 (1.30–4.94)]. This study revealed that both IL-33 SNPs were associated with an increased risk of asthma. The rs1342326 was associated with atopic, mild and adult-onset asthma and a higher level of eosinophils in peripheral blood. However, rs3939286 was more frequent in moderate and severe asthma. Moreover, rs3939286 was associated with non-atopic and childhood-onset asthma.

Nazanin Zahrooni,1 Seyed Ahmad Hosseini,2 Ahmad Ahmadzadeh,3 Kambiz Ahmadi Angali,4 Mohammad Ali Assarehzadegan5 1Department of Nutrition, Faculty of Paramedicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 2Nutrition and Metabolic Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 3Thalassemia and Hemoglobinopathy Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 4Biostatistics Division, Health School, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 5Department of Immunology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IranCorrespondence: Seyed Ahmad HosseiniNutrition and Metabolic Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IranTel +98-6133367543Fax +98-6133720299Email Seyedahmadhosseini@yahoo.comBackground: To better evaluate the efficacy of CoQ10 on the inflammatory markers in breast cancer patients, we conducted a clinical study of patients with breast cancer undergoing tamoxifen therapy. CoQ10 serves as an antioxidant and inhibits oxidation caused by reactive oxygen species. The aim of the current study was to assess the effect of coenzyme Q10 supplementation on serum levels of interleukin 6, 8, and vascular endothelial growth factor (VEGF) in patients with breast cancer undergoing tamoxifen therapy by a double-blind, placebo-controlled, randomized clinical trial.Methods: In the study, 30 breast cancer patients and 29 healthy subjects were randomized into four groups. Two groups of intervention received 100 mg CoQ10, and two control groups took placebo once a day for 2 months. Blood draws were obtained at baseline and at the end of the study. Serum levels of IL-6, IL-8 and VEGF were analyzed using ELISA kits.Results: The data of the 59 participants were analyzed. Supplementation with CoQ10 demonstrated a significant decrease in IL-8 and IL-6 serum levels compared to placebo (P< 0.05). Although the downward trend was evident, CoQ10 supplementation did not reveal any significant effect on serum VEGF concentration. The group of patients who received supplements showed the most reduction in serum levels of cytokines among other groups.Conclusion: CoQ10 supplementation could be effective in ameliorating inflammatory cytokine levels, thereby reducing the consequences of inflammation caused by breast cancer. To generalize the results, larger and longer intervention studies with higher safe doses are needed and should take account of possible costs and harms as well as benefits (registration number: IRCT2015042021874N1).Keywords: breast cancer, CoQ10, inflammatory cytokines, quality of life, tamoxifen

A disastrous outbreak of thunderstorm-induced asthma attacks posed a major public health threat in Ahvaz, Iran in autumn of 2013 (>15,000 referrals to hospital emergency rooms and pulmonary clinics). City officials claimed that promoting factors included acid rain, particulate matter, ozone, and polycyclic aromatic hydrocarbon compounds (PAHS) could lead to this phenomenon. After these potential causes were rejected by careful research, the next possible cause that this study addresses is a potential link of the asthma attacks to airborne pollen grains and fungal spores. In order to sample these parameters, a Hirst-type volumetric spore sampler was used, which was installed on the rooftop of the sampling site. Sampling was done during two six-month periods to characterize the regional bioaerosol. The first period included the autumn (September 22 to December 20 of 2016) and winter seasons (December 21 of 2016 to March 19 of 2017), while the second period included the following autumn (September 23 to December 21 of 2017) and winter seasons (December 22 of 2017 to March 20 of 2018). In both studied periods, airborne weed pollen grains, especially Amaranthaceae sp., were by far the greatest contributor to total airborne pollen grain concentrations (50.45%, 3757 pollen grains/m3 and 52.12%, 2392 pollen grains/m3, respectively). Among fungal spores, Cladosporium had the highest concentrations (75.29%, 258677 spores/m3 and 80.33%, 247693 spores/m3, respectively). Findings also showed that concentrations of airborne pollen grains were higher in autumn seasons (6361 and 4167 pollen grains/m3, respectively) in comparison with winter seasons (1085 and 423 pollen grains/m3, respectively), while the concentrations of fungal spores were enhanced in winter seasons (189216 and 160962 spores/m3, respectively) versus autumn seasons (154377 and 147377 spores/m3, respectively). Fungal spores exhibited a uniform distribution during the entire study period, but airborne pollen grains were significantly higher in autumn seasons. The numbers of referrals associated with thunderstorm asthma attacks decreased drastically after implementing Conocarpus erectus tree pruning activities. We concluded that the autumnal thunderstorm asthma attacks in Ahvaz might be triggered by a combination of bio-allergens (fungal spores and airborne pollen grains such as from Conocarpus erectus) and high levels of air pollutants from industrial activity. The results of this work suggest controlling bio-allergen levels by preventive measures such as tree pruning, weed removal, and emasculating urban green infrastructures before the flowering season.

Introduction:Little is known about the laboratory and radiological characteristics and clinical significance of peripheral immune alterations in patients with coronavirus disease 2019 (COVID-19). This study aims to clarify these aspects in children and adults with COVID-19.Methods:In this consecutive pilot study, COVID-19 patients with the confirmed pneumonia and real-time RT-PCR were recruited prospectively in June 2020. The clinical, chest CT, and laboratory features, such as lymphocyte subpopulations, were analyzed for each individual.Results:Forty confirmed COVID-19 patients, 11 severe children, 12 severe adults, and 17 critical adult patients, besides 20 healthy pediatrics and 14 healthy adults as controls, were enrolled prospectively. Adult patients, especially critical ones, had a much higher prevalence of laboratory and chest CT abnormalities. Data regarding immune cell subsets in children patients, compared with matched controls, had higher CD3+ CD8+ T cells (p= 0.004) and lower CD4+/CD8+ ratio (p= 0.042), while adult patients, compared with matched controls, had lower CD14+ monocytes (p= 0.032). Adult patients were also categorized as experiencing critical or severe illness on admission and, compared with severe patients, had lower total lymphocytes (p< 0.047), CD3+ T-lymphocytes (p< 0.002), and CD3+ CD8+ T cells (p= 0.001) and, on the other hand, had higher CD3+ CD4+ T cells (p= 0.012) and CD4+/CD8+ ratio (p= 0.003). Non survived adults, compared with survived patients, had significantly lower CD3+ T-lymphocyte (p= 0.005).Conclusion:Unlike adult patients, who compared with matched controls and had more comorbidities, higher frequency of severe clinical symptoms, laboratory abnormalities, and immune cells alteration, clinical manifestations of COVID-19 in children (compared with matched controls) were relatively mild, and fewer clinical complications were seen either, perhaps because of a milder inflammatory response following their peripheral innate and adaptive immune cell alteration pattern.

Background: Overexpression of miR-21 is a characteristic feature of patients with Multiple Sclerosis (MS) and is involved in gene regulation and the expression enhancement of pro-inflammatory factors including IFNγ and TNF-α following stimulation of T-cells via the T Cell Receptor (TCR). In this study, a novel integrated bioinformatics analysis was used to obtain a better understanding of the involvement of miR-21 in the development of MS, its protein biomarker signatures, RNA levels, and drug interactions through existing microarray and RNA-seq datasets of MS. Methods: In order to obtain data on the Differentially Expressed Genes (DEGs) in patients with MS and normal controls, the GEO2R web tool was used to analyze the Gene Expression Omnibus (GEO) datasets, and then Protein-Protein Interaction (PPI) networks of co-expressed DEGs were designed using STRING. A molecular network of miRNA-genes and drugs based on differentially expressed genes was created for T-cells of MS patients to identify the targets of miR-21, that may act as important regulators and potential biomarkers for early diagnosis, prognosis and, potential therapeutic targets for MS. Results: It found that seven genes (NRIP1, ARNT, KDM7A, S100A10, AK2, TGFβR2, and IL-6R) are regulated by drugs used in MS and miR-21. Finally, three overlapping genes (S100A10, NRIP1, KDM7A) were identified between miRNA-gene-drug network and nineteen genes as hub genes which can reflect the pathophysiology of MS. Conclusion: Our findings suggest that miR-21 and MS-related drugs can act synergistically to regulate several genes in the existing datasets, and miR-21 inhibitors have the potential to be used in MS treatment.

Ligustrum vulgare (Privet) pollen proteins are responsible for allergies in susceptible individuals in many regions of the world. This study investigated the immunochemical characterization of Privet pollen extract and the occurrence of skin prick test reactivity to Privet and other allergenic pollen grains in allergic rhinitis patients. All subjects experienced a skin prick test with twenty-two allergen extracts. sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) separated Privet pollen extract, IgE-immunoblotting, and specific ELISA procedures determined the allergenic profile on forty-five Privet allergic patients. A positive allergic reaction to L. vulgare pollen extract was observed in forty-five (31.4%) out of 145 patients. Ten resolved protein fractions were found on SDS-PAGE, ranging from 10 to 80 kDa. IgE-specific antibodies interacted with several allergenic protein bands from Privet-allergic patients in the immunoblotting assay. The most significant interaction was observed in proteins with molecular weights of approximately 15, 18, 43, and 66 kDa. Privet pollen is regarded as a potent allergen composed of IgE-binding constituents. Considering the high allergenicity of Privet pollen grains and since many countries are rich in this plant, identification and production of recombinant forms of common allergens in this species can be used for developing more efficient diagnostic, therapeutic, and preventive approaches.

Leukemia inhibitory factor (LIF) is a multi-functional cytokine secreted from cells such as lymphocytes and hepatocytes. This study aimed to evaluate the effect of LIF on natural killer group 2 member D (NKG2D) receptors' expression and presentation on natural killer (NK) cells. For this purpose, peripheral blood mononuclear cells taken from 4 young male healthy blood donors were isolated and the effect of LIF (25 ng/mL) after 12, 24, and 48 hours of incubation, on NKG2D receptors expression and presentation was investigated using flow cytometry and real-time-polymerase chain reaction (PCR). All of the steps of the experiment were performed in duplicate. After periods of 12, 24, and 48 hours, LIF reduced both the expression and presentation of the NKG2D receptor on NK cells. The results suggest that this cytokine has a direct modulating activity on the body’s immune response through suppression of NKG2D receptor expression and presentation on NK cells.

Bevacizumab with anti-angiogenesis properties reduces the vascular endothelial growth factor (VEGF) level and has widely been used to treat various diseases such as lung diseases and chronic obstructive pulmonary disease (COPD). This study, therefore, aimed to consider the effects of bevacizumab on VEGF receptor 2 (VEGFR2) and lung inflammation of the ovalbumin-induced rat model of airway hypersensitivity. Twenty-one male Wistar rats were randomly divided into 3 groups (n = 7 in each group): (1) control, (2) ovalbumin (OVA)-sensitized, and (3) OVA-sensitized with bevacizumab (OVA + Bmab). Groups 2 and 3 were sensitized with ovalbumin (OVA) and aluminum hydroxide on days 1, 8 and challenged with OVA on day 15 by atomization for 10 days (inhalation). After OVA sensitization, the OVA + Bmab was treated with bevacizumab for 2 weeks. VEGFR2 was semiquantitatively analyzed in the lungs by immunohistochemistry. VEGF was measured in the lung tissue by ELISA method. The mRNA of IL-10 and IL-6 lung tissue were measured by real-time PCR. Ovalbumin exposure promoted the expression of VEGF and resulted in inflammatory factors overexpression (p ≤ 0.05). However, rats in OVA + Bmab group showed significantly a decrease in VEGFR2 and IL-1β, IL-6, TNFα, and an increase in IL-10 (p ≤ 0.05). The results show that bevacizumab efficiently diminishes bronchial inflammation via reducing the expression of VEGFR2, and IL-6 genes and enhancing the expression of IL-10 gene. Hence, bevacizumab could be considered as a potential candidate drug to control pathological conditions relevant to airway hypersensitivity.

— Studies on the bronchial vascular bed have revealed that the number of blood vessels in the lamina propria and under the mucosa of the lung tissue increases in patients suffering from mild to severe asthma. Thus, in this study, a new strategy was employed in respiratory system disorders by angiogenesis inhibition in an ovalbumin (OVA)-induced rat model of asthma. Twenty-one male Wistar albino rats, 8 weeks old, were randomly divided into three groups (n = 7 in each group), including (1) control group, (2) OVA-treated group, and (3) OVA + Bmab (bevacizumab drug). On days 1 and 8, 1 mg of OVA and aluminum hydroxide in sterile phosphate-buffered saline (PBS) were intraperitoneally injected to rats in groups 2 and 3. The control group was only subject to intraperitoneal injection of saline on days 1 and 8. One week after the last injection, the rats (groups 2 and 3) were exposed to OVA inhalation for 30 min at 2-day intervals from days 15 to 25. After sensitization and challenge with OVA, the OVA + Bmab group (group 3) were treated with a 5 mg/kg bevacizumab drug. Genes and protein expression of IL-1β and TNF-α and the expression of vascular endothelial growth factor (VEGF) protein were assessed by real-time PCR and immunohistochemistry respectively, in lung tissue. OVA exposure increased mucosal secretion and inflammatory cell populations in lung tissue and OVA-specific IgE level in serum. Also, VEGF and cytokine factor expression were significantly elevated in the OVA-induced asthma model (p ≤ 0.05). However, rats in OVA + Bmab group showed significantly a decrease in VEGF and IL-1β and TNF-α genes as well as proteins (p ≤ 0.05). The results showed that bevacizumab efficiently diminished bronchial inflammation via downregulation of VEGF expression, followed by inflammatory cells population and cytokines reduction. Angiogenesis inhibition in rats with induced asthma not only suppresses the inflammatory process through blocking VEGF expression but also inhibits the development of new blood vessels and progressing asthmatic attacks.

Long-term exposure to airborne particles of 10 µm and less in size (PM(10)) in dust can lead to increased risk of diseases such as respiratory, cardiovascular, lung cancer and atherosclerosis. The aim of the study was to evaluate the effects of water-soluble PM(10) particles in the Middle East Dust (MED) storm in Ahvaz, Iran, on the production of TNF-α by human monocytes. In addition, we assessed the level of induction of apoptosis in isolated A549 human pulmonary epithelial cells. For this purpose, isolated human blood monocytes (250,000 to 300,000 cell/ ml) as well as isolated human pulmonary A549 epithelial cells (100,0000 cell/ ml) were exposed to various concentrations (62.5, 125, 250, 500 µg/ml) of water-soluble PM(10) particles for different incubation periods (12, 24, 48 h). The results showed that exposure to PM(10) particles increased the production of TNF-α in human monocytes and promoted apoptosis induction in A549 cells, in both concentration and incubation of period-dependent manner. In conclusion, airborne dust particles in Ahvaz city contain active compounds capable of increasing production of the pro-inflammatory cytokine, TNF-α, and inducing apoptosis of lung epithelial cells.

Natural killer (NK) cells are a subset of lymphocytes in humans that release cytokines such as tumor necrosis factor alpha and interferon gamma-γ during infection. NKG2D is one of the most important stimulating NK receptors binding MIC-A, MIC-B, and ULBPs, which leads to activation of NK cells against tumor cells. In this study, the authors evaluated the effect of G2 adjuvant on gene expression and delivery of NKG2D receptor on NK cells in peripheral blood.Peripheral blood mononuclear cells were isolated from venous blood obtained from healthy volunteers after adding G2 adjuvant within 12, 24, and 48 hours of incubation. Then, total RNA was extracted from the cells, cDNA synthesis was performed, and gene expression was evaluated by real-time PCR. In addition, NK cells were stained with the appropriate monoclonal antibodies, and the receptors expressed on cell surface were quantified.G2 adjuvant leads to upregulation of gene expression and increases the expression of NKG2D receptor on the surface of NK cells after incubation.The findings of this study demonstrated that G2 adjuvant can increase NK cell cytotoxicity. It may play an important role in killing tumor cells, preventing tumor growth and metastasis.

Acacia farnesiana is the main source of allergenic pollen and one of the most important causes of respiratory allergic disease in tropical and subtropical regions of the world. The purpose of this study was to produce a recombinant variety of allergenic Ole e 1-like protein from the pollen of this tree. To predict its allergenic cross-reactivity with other members of the Ole e 1-like protein family of common allergenic plants, the nucleotide sequence homology of the Acacia Ole e 1-like protein was evaluated. Amplification of cDNA strands encoding Acacia Ole e 1-like protein was performed by polymerase chain reaction (PCR) and sequenced. Following expression in Escherichia coli using the pET-21b(+) vector, the recombinant protein was purified using metal-affinity chromatography. IgE-binding competence of purified recombinant Ole e 1- like protein (rAca f 1) was analysed by immunoassay using 25 sera collected from Acacia pollen-sensitised patients. Nucleotide sequencing revealed an open reading frame of 453 bp encoding 150 amino acid residues that belonged to the Ole e 1-like protein family, and 11 patients (44%) had considerable specific IgE levels for the rAca f 1. Immunodetection and inhibition assays indicated that the purified rAca f 1 may be the same as that in the crude extract. Aca f 1, the second allergen from Acacia pollen, was identified as a member of the family of Ole e 1-like protein. A high degree of homology was found among amino acid sequences of Aca f 1 and several allergenic members of Ole e 1-like protein family.

Kochia scoparia pollen has been demonstrated as an important cause of pollinosis in tropical and sub-tropical regions of the world. The aim of this study was to characterize the IgE-binding protein of the Kochia pollen extract and production of recombinant form of allergenic profilin of this weed. To predict its allergenic cross-reactivity with profilins of common allergenic plants, nucleotide sequence homology of Kochia profilin was evaluated. Specific ELISA and immunoblotting assay were applied to determine the IgE-binding reactivity of 28 sera collected from patients who were sensitized to Kochia pollen. In cloning procedure, the Kochia profilin-coding sequence was inserted into PTZ57R/T vector and expressed using pET-21b(+) vector. IgE-binding competence of purified recombinant Kochia profilin (rKoc s 2) was analyzed by in vitro assays. There were several protein bands in Kochia pollen extract with molecular weights approximately ranging from 14.5 to 85 kDa. Nucleotide sequencing revealed an open reading frame of 399 bp encoding for 133 amino acid residues which belonged to the profilin family; 15 patients (15/28, 53.1 %) had significant specific IgE levels for the rKoc s 2. Immunodetection and inhibition assays indicated that the purified rKoc s 2 might be the same as that in the crude extract. Koc s 2, the first allergen from the Kochia pollen was identified as a member of the family of profilins. High degree of homology was found among amino acid sequences of Kochia profilin and several profilin molecules from unrelated plant families.

Background: Today, leukemia is one of the biggest problems worldwide. The Wilms’ tumor gene (WT1) and the vascular endothelial growth factor (VEGF) gene are highly expressed in patients with various cancers. This study concerned the relationship between expression of WT1 and VEGF in patients with acute leukemia. Materials and Methods: We evaluated expression of WT1 mRNA and VEGF mRNA using real-time quantitative RT-PCR in the peripheral blood (PB) of 8 newly diagnosed AML and 4 newly diagnosed ALL patients, serially monitored for 2 months. A further 12 normal PB samples served as controls. Results: In the patient group, in comparison with the normal ranges, WT1 and VEGF gene expression was increased, the average values for the expression of these two genes being 0.2852±0.11 and 0.2029±0.018, respectively. While was no significant relevance between the two genes pre-treatment, a positive link between the two genes in 75% of patients with AML was noted during the procedure of chemotherapy, whereas in 75% of patients with ALL an antiparallel association was observed. Conclusions: Leukemia is associated with production of WT1, which may affect the expression of VEGF.

Hemiscorpius lepturus (H. lepturus), one of the most venomous scorpions in tropical and sub-tropical areas, belongs to the Hemiscorpiidae family. Studies of antibodies in sera against the protein component of the venom from this organism can be of great use for the development of engineered variants of proteins for eventual use in the diagnosis/treatment of, and prevention of reactions to, stings. In the present in vitro study, the proteins of H. lepturus venom, which could specifically activate the production of immunoglobulin G (IgG) in victims accidently exposed to the venom from this scorpion, were evaluated and their cross-reactivity with venoms from two other important scorpion species including Androctonus crassicauda and Mesobuthus eupeus assessed. H. lepturus venom was analyzed with respect to its protein composition and its antigenic properties against antibodies found in sera collected from victims exposed to the venom of this scorpion within a previous 2-month period. The cross-reactivity of the H. lepturus venom with those from A. crassicauda and M. eupeus was assessed using ELISA and immunoblotting. Electrophoretic analysis of the venom of H. lepturus revealed several protein bands with weights of 8-116 KDa. The most frequent IgG-reactive bands in the test sera had weights of 34, 50, and 116 kDa. A weak cross-reactivity H. lepturus of venom with venoms from A. crassicauda and M. eupeus was detected. The results of immunoblotting and ELISA experiments revealed that H. lepturus venom activated the host immune response, leading to the production of a high titer of antibodies. Clearly, a determination of the major immunogenic components of H. lepturus venom could be valuable for future studies and ultimately of great importance for the potential production of recombinant or hypo-venom variants of these proteins.

The aim of this study was to report the characteristics and treatment strategies of all patients with acute bronchospasm who were presented to the emergency departments of Ahvaz, Iran, following the occurrence of a thunderstorm on November 2, 2013. A total of 2000 patients presenting with asthma attacks triggered by thunderstorm were interviewed and an initial questionnaire was completed for each individual. After twenty days, patients were asked to complete a supplementary questionnaire, but only 800 of them accepted to do so. The majority of subjects was aged 20–40 years (60.5%) and had no history of asthma in most cases (60.0%). The symptoms had started outdoors for 60.0% of the participants. In most patients, the onset of the condition was on November 2. Short-actingβ2-agonist (salbutamol) and aminophylline were the most commonly prescribed medications in the emergency department. Upon the second interview, 85.3% of the patients were still symptomatic. Overall, 63.6% did not have a follow-up visit after hospital discharge, although all of them were referred to the specialist. The findings of the present study suggest that thunderstorm-associated asthma could affect young adults with no gender priority, with or without asthma history, which put a strain on emergency medical services.

Background. Vitis vinifera vitis (grape) cultivar Sultana is a causative agent of oral allergy syndrome (OAS) in Iran. In this study, the IgE-reactive proteins of this cultivar were characterised. Methods. IgE-reactivity of sera from patients with history of OAS to grape and positive skin prick test (SPT) were evaluated. Grape immunoreactive components were also purified and characterised. Results. The purified 10 kDa allergen showed the highest immunoreactivity in component-resolved ELISA and the results were in correlation with SPT results. The patients' sera were mostly reactive with 10, 28 and 60 kDa allergens; meanwhile, a 16 kDa protein can be considered as a novel marker for OAS to grape. The 24, 28 and 30 kDa allergens were identified as thaumatin-like protein, class IV chitinase and beta-1,3-glucanase, respectively. Conclusion. The Sultana cultivar showed a rather similar pattern of IgE reactivity with other cultivars. Several allergens play role in OAS to grapes, however, the 10 kD protein is th...

Background: Pulmonary fibrosis (PF) is among the world’s most prevalent and common respiratory system diseases. Several previous studies have revealed the contribution of angiotensin in the pathogenesis of PF; subsequently, angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin receptor antagonists (ARDs) have been proposed for the treatment of pulmonary fibrosis using therapeutic approaches. Objectives: This study aimed to investigate the anti-fibrotic effect of valsartan as an angiotensin receptor blocker. Methods: Rats were given a single intratracheal administration of bleomycin (7.5 IU/kg); valsartan (20, 40, 80 mg/kg/day) was administrated to the rats orally, starting seven days before the induction of lung fibrosis and continuing until the end of the study. The control group received a vehicle. Results: The rats that received valsartan exhibited decreased hydroxyprolin content and pulmonary index values. Pathological examination showed that valsartan could prevent inflammation and fibrotic scarring induced by bleomycin. Conclusions: Valsartan displayed an anti-fibrotic and protective effect against bleomycin-induced lung fibrosis in an animal model.

Pollen from mesquite (Prosopis juliflora) is one of the important causes of immediate hypersensitivity reactions in the arid and semi-arid regions of the world. The aim of present study is to produce and purify the recombinant form of allergenic Ole e 1-like protein from the pollen of this allergenic tree. Immunological and cross-inhibition assays were performed for the evaluation of IgE-binding capacity of purified recombinant protein. For molecular cloning, the coding sequence of the mesquite Ole e 1-like protein was inserted into pTZ57R/T vector and expressed in Escherichia coli using the vector pET-21b(+). After purification of the recombinant protein, its immunoreactivity was analysed by in vitro assays using sera from twenty one patients with an allergy to mesquite pollen. The purified recombinant allergen was a member of Ole e 1-like protein family and consisted of 150 amino acid residues, with a predicted molecular mass of 16.5 kDa and a calculated isoelectric point (pI) of 4.75. Twelve patients (57.14%) had significant specific IgE levels for this recombinant allergen. Immunodetection and inhibition assays indicated that the purified recombinant allergen might be the same as that in the crude extract. Herein, we introduce an important new allergen from P. juliflora pollen (Pro j 1), which is a member of the Ole e 1-like protein family and exhibits significant identity and similarity to other allergenic members of this family.

Background Pollinosis from Amaranthus retroflexus pollen is a common cause of respiratory allergy in Iran with a high positive rate (68.8%) among Iranian allergic patients. The aim of the present study was to evaluate the allergenicity of the A. retroflexus pollen profilin. Methods Using sera from twelve patients allergic to A. retroflexus pollen, IgE-binding proteins from the A. retroflexus pollen extract was identified by immunoblotting. The cDNA of A. retroflexus pollen profilin was amplified, then cloned into the pET-21b (+) vector, expressed in Escherichia coli, and finally purified by metal affinity chromatography. The IgE-binding capacity of the recombinant protein was then analyzed by the ELISA, immunoblotting, and inhibition assays, as well as by the skin prick test (SPT). Results Immunoblotting results indicated a 14.6 kDa protein with IgE-reactivity to 33% (4/12) among A. retroflexus pollen-allergic patients. Nucleotide sequencing of the cDNA revealed an open reading frame of 399 bp encoding for 133 amino acid residues which was belonged to the profilin family and designated as Ama r 2. A recombinant Ama r 2 (rAma r 2) was then produced in E. coli as a soluble protein which showed a strong IgEreactivity via ELISA confirmed by the SPT. Inhibition experiments revealed high IgE cross-reactivities with the profilins from other plants. Conclusions The profilin from the A. retroflexus pollen, Ama r 2, was firstly identified as an allergen. Moreover, rAma r 2 was produced in E. coli as a soluble immunoreactive protein with an IgE-reactivity similar to that of its natural counterpart.

The inhalation of Chenopodium album (C. album) pollen has been reported as an important cause of allergic respiratory symptoms. The aim of this study was to produce the recombinant profilin of C. album (rChe a 2) pollen and to investigate its cross-reactivity with other plant-derived profilins based on potential conformational epitopes and IgE reactivity analysis. Che a 2-coding sequence was cloned, expressed, and purified using one step metal affinity chromatography to recover high-purity target protein. We assessed cross-reactivity and predicted IgE potential epitopes among rChe a 2 and other plant-derived profilins. Immunodetection and inhibition assays using sixteen individual sera from C. album allergic patients demonstrated that purified rChe a 2 could be the same as that in the crude extract. The results of inhibition assays among rChe a 2 and other plant-derived profilins were in accordance with those of the homology of predicted conserved conformational regions. In this study, amino acid sequence homology analysis showed that a high degree of IgE cross-reactivity among plant-derived profilins may depend on predicted potential IgE epitopes.

Salsola kali pollen is a common cause of pollinosis during summer and early fall in desert and semi-desert regions. The aim of this study was the identification and characterization of Sal k 3, a new allergen from S. kali pollen. S. kali pollen extract was fractionated by SDS-PAGE and the allergenic profile was determined by IgE-immunoblotting using twelve S. kali allergic patients. Protein identification was carried out by the means of mass spectrometry. Using degenerated primers, two DNA fragments encoding N- and C-terminal domain of Sal k 3 were amplified by PCR, then cloned into the PTZ57R/T vector and sequenced. The open reading frame of Sal k 3 fragments were subcloned in the pET-32b(+) vector, expressed in E. coli, and purified by Ni2+ affinity chromatography. The IgE-binding capacity of rSal k 3 fragments was then studied by IgE-immunoblotting, inhibition assays, and skin prick tests. A 45-kDa allergen was identified as a fragment of the cobalamin-independent methionine synthase (MetE) by mass spectrometry and was detected in the sera of 8/12 (66.6%) of S. kali allergic patients. Moreover, inhibition assays demonstrated that the purified rSal k 3 fragments were similar to their counterparts in the crude extract. Sal k 3 represents a new allergen of S. kali pollen and seems to be an important allergenic compound in S. kali pollen.

Background Pollens from the Salsola spp. are an important source of respiratory allergy in tropical countries. Our aim was to characterize the IgE binding proteins of S. incanescens pollen extract and study its cross-reactivity with S. kali pollen allergens. Methods Prick tests with S . kali and S . incanescens pollen extracts were performed on eight respiratory allergy patients from Mashhad, Northeast Iran. The antigenic profiles and IgE-binding patterns of S. kali and S. incanescens pollen extracts were compared by SDS-PAGE and Western blotting, using individual sera from the salsola pollen-sensitive patients. Cross-reactivity of proteins in the two weeds was assessed by IgE- immunoblotting inhibition. Results S. kali and S. incanescens pollen extracts showed similar IgE-binding profiles in Western blotting. The IgE binding components of 39, 45, 66 and 85 kDa were detected in both pollen extracts. Furthermore, inhibition of the immunoblots revealed extensive inhibition of IgE binding to proteins and a close relationship between these two weeds allergens. Conclusions S. incanescens pollen is a potent allergen source with several IgE binding components that shows a close allergenic relationship with S. kali . Our results suggest that in S. incanescens - rich areas, S. kali pollen extracts could be used as a diagnostic reagent for allergic patients to S. incanescens pollen.