Payam Noor University of Mashhad

of Mashhad, Iran

Payam Noor University of Mashhad

of Mashhad, Iran
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Mosadeghi P.,Payam Noor University of Mashhad | Zarnagh H.H.,Sabzevar University of Medical Science | Mohammad-Zadeh M.,Sabzevar University of Medical Science | Moghaddam M.S.,Payam Noor University of Mashhad
Iranian Journal of Allergy, Asthma and Immunology | Year: 2015

Expression of HTLV-I p19 protein in an Escherichia coli expression system always leads to the formation of inclusion body. Solubilisation and refolding of the inclusion bodies is complex, time consuming and difficult during large-scale preparation. This study aimed to express and purify a soluble form of recombinant HTLV-I p19 protein in an E. coli expression system. The synthetic DNA encoding the p19 was subcloned into a pGS21a vector along with a His-GST solubility/purification tag. The recombinant pGS21a-p19 vector was then transformed into chemically competent E. coli BL21 (DE3) cells, and expression of the recombinant His-GST-p19 protein was induced by IPTG. Expression and distribution of the His-GST-p19 protein in soluble and insoluble fractions were evaluated using SDS-PAGE. Antigenicity of the His-GST-p19 protein was evaluated using ELISA after purifying the protein using Ni-NTA affinity chromatography, then compared to the results of synthetic immunodominant p19 peptide ELISA. The fusion His-GST-p19 protein accounted for 30% of the total cellular proteins. The SDS-PAGE results indicated that approximately 50% of the expressed His-GST-p19 proteins were soluble and accounted for 50% of the total soluble proteins. ELISA showed that the His-GST tag did not impair the antigenicity of the p19 protein and that the fusion protein reacted with HTLV-I antibodies in a concentration-dependent manner. The results of His-GST-p19 ELISA indicated that specificity of p19 reactivity was compatible to the results of p19 peptide ELISA. Combination of key strategies for the soluble expression of proteins, like fusion with solubility/purification tags, low IPTG concentration and induction at low temperature, provide an efficient and facile platform for producing soluble HTLV-I p19 protein. © Autumn 2015, Iran J Allergy Asthma Immunol. All rights reserved.


PubMed | Sabzevar University of Medical Science and Payam Noor University of Mashhad
Type: Journal Article | Journal: Iranian journal of allergy, asthma, and immunology | Year: 2016

Expression of HTLV-I p19 protein in an Escherichia coli expression system always leads to the formation of inclusion body. Solubilisation and refolding of the inclusion bodies is complex, time consuming and difficult during large-scale preparation. This study aimed to express and purify a soluble form of recombinant HTLV-I p19 protein in an E. coli expression system. The synthetic DNA encoding the p19 was subcloned into a pGS21a vector along with a His-GST solubility/purification tag. The recombinant pGS21a-p19 vector was then transformed into chemically competent E. coli BL21 (DE3) cells, and expression of the recombinant His-GST-p19 protein was induced by IPTG. Expression and distribution of the His-GST-p19 protein in soluble and insoluble fractions were evaluated using SDS-PAGE. Antigenicity of the His-GST-p19 protein was evaluated using ELISA after purifying the protein using Ni-NTA affinity chromatography, then compared to the results of synthetic immunodominant p19 peptide ELISA. The fusion His-GST-p19 protein accounted for 30% of the total cellular proteins. The SDS-PAGE results indicated that approximately 50% of the expressed His-GST-p19 proteins were soluble and accounted for 50% of the total soluble proteins. ELISA showed that the His-GST tag did not impair the antigenicity of the p19 protein and that the fusion protein reacted with HTLV-I antibodies in a concentration-dependent manner. The results of His-GST-p19 ELISA indicated that specificity of p19 reactivity was compatible to the results of p19 peptide ELISA. Combination of key strategies for the soluble expresion of proteins, like fusion with solubility/purification tags, low IPTG concentration and induction at low temperature, provide an efficient and facile platform for producing soluble HTLV-I p19 protein.


Jamialahmadi K.,Mashhad University of Medical Sciences | Sadeghnia H.R.,Mashhad University of Medical Sciences | Mohammadi G.,Payam Noor University of Mashhad | Kazemabad A.M.,Islamic Azad University at Tehran | Hosseini M.,Mashhad University of Medical Sciences
Pathophysiology | Year: 2013

Introduction and objective: Beneficial effects of glucosamine in spatial learning and memory impairment induced by scopolamine has been evaluated in rats by using Morris water maze. Methods: Male Wistar rats were randomly divided into control, scopolamine and scopolamine plus glucosamine groups. All injections were given in 5 consecutive days and 30. min after each injection, the rats were tested in the Morris water maze test. Escape latency and path length to reach the hidden platform were subjected to analysis of variance [ANOVA]. Results: The rats treated with scopolamine showed increased escape latency and path length to reach the hidden platform compared to control group (P< 0.001). Both escape latency and traveled path length to reach the hidden platform in glucosamine treated animals (1 and 2. g/kg) were significantly lower (P< 0.05 to P<. 0.001) than in the scopolamine group. Conclusion: The results of this study showed that the glucosamine can inhibit scopolamine-induced impairments of spatial learning and memory in rats. Glucosamine might offer a promise in either the prevention or the treatment of neurodegenerative diseases such as Alzheimer's disease. © 2013 Elsevier Ireland Ltd.


Abbasnejad E.,Education 3rd District Mashhad | Farahani A.,University of Tehran | Nakhaei A.,Payam Noor University of Mashhad
Advances in Environmental Biology | Year: 2013

The present study was carried out to investigate about the relationship between time management and jobrelated stress. For this purpose, 625 teachers working in the education department of Mashhad were selected as samples. These subjects filled time management questionnaire with reliability of 0.86, and job stress questionnaire with reliability of 0.74. Results of Kolmogorov-Smirnov test, Pearson correlation test, 2-string Spirman test, t-test and Mann-Whitney U test showed that: - There is a difference between time management in physical and non-physical education teachers, but it's not a significant difference. Time management of non-PE teachers is higher. - There is a difference between job stress of PE and non-PE teachers, but it's not a significant difference. Job stress of PE teachers is higher than of the other group. - There is a significant but inverse relationship between time management and job stress. - There is no significant relationship between job stress and age, gender, work experience and education level in non-physical education teachers. - There is a direct and significant relationship between time management and age and work experience (in contrast with gender and education level) in PE and non-PE teachers.


Minai-Tehrani D.,Shahid Beheshti University | Masoudnia A.,Payam Noor University of Mashhad | Alavi S.,Shahid Beheshti University | Osmani R.,Shahid Beheshti University | And 4 more authors.
Drug Metabolism and Drug Interactions | Year: 2012

Background: Methocarbamol is a skeletal muscle relaxant and is widely used to relieve pain in muscles. Many drugs may have interactions with each other when used at the same time. Yeast sucrase is taken as a drug by patients with congenital sucrase-isomaltase deficiency (CSID). Methods: In this study, the interaction between methocarbamol and yeast sucrase was investigated. Results: Our results showed that methocarbamol can inhibit sucrase activity and reduce the maximum reaction velocity (Vmax) of the enzyme by a non-competitive pattern. Measurement of IC50 and Ki of the drug revealed that methocarbamol did not bind the enzyme with high affinity. Fluorescence measurement showed that the drug binds to free enzyme and enzyme-substrate complexes that were accompanied by structural changes on the enzyme. Guaifenesin, which has a similar structure to methocarbamol, does not affect the activity of sucrase. Conclusions: Methocarbamol inhibits sucrase activity and its carbamate group plays a main role in the binding of drug to sucrase.


Malvandi A.M.,Payam Noor University of Mashhad | Mehrzad J.,Ferdowsi University of Mashhad | Saleh-Moghaddam M.,Payam Noor University of Mashhad
Immunopharmacology and Immunotoxicology | Year: 2013

Context: Aflatoxins (AFs) are highly hazardous carcinogenic mycotoxins originated from very common fungi present in the environment. Their effect on key immune-surveillance molecules is unclear. Objective: We aimed to examine the effect of mixed AFs on immunologically relevant molecules and on viability in human peripheral blood mononuclear cells (PBMCs), in conditions similar to those occurring naturally, i.e. using a mixture of environmentally relevant levels of AFB1, AFB2, AFG1 and AFG2. Materials and methods: We evaluated the mRNA expression of MyD88, toll-like receptor (TLR)-2, TLR4 and CD14, in human PBMCs treated with a mixture of AFB1, AFB2, AFG1 and AFG2 at different doses for 2, 12 and 24th. We used qRT-PCR to assess changes in transcripts of MyD88, TLR2, TLR4 and CD14 in PBMCs. We also evaluated the viability of PBMCs exposed to AFs. Results: Biologically relevant levels of mixed AFs elicited early immune modulation in human PBMCs. qRT-PCR results showed several folds increase of MyD88, TLR2, TLR4 and CD14 transcripts in PBMCs as early as 2th post-exposure to mixed AFs. Kinetics and dose-response of the up-regulation differed for mentioned gene transcripts. Further, prolonged exposure to mixed AFs decreased PBMCs viability. Conclusion: Immunotoxicity of AFs on PBMCs may be mediated by up-regulation of key immune-surveillance molecule transcripts. The description of these effects induced by AFs on PBMCs are novel and should be taken into account when considering AF-related infectious and noninfectious diseases in areas highly exposed to AFs. © 2013 Informa Healthcare USA, Inc. All rights reserved: reproduction in whole or part not permitted.


Mohammadi G.,Payam Noor University of Mashhad
Pakistan journal of biological sciences: PJBS | Year: 2011

The aim of this study was to investigate the expression of membranous epidermal growth factor receptor in colorectal adenocarcinoma and it's correlation with clinicopathological features. Fifty formalin-fixed, paraffin embedded archival specimens of colorectal cancer were included randomly as cases. Immunohistochemical staining was performed to assess EGFR expression. The results were correlated with the clinicopathological features of colorectal tumor tissues. More than 1% of membranous EGFR expression was found in 24 (48%) of cancer specimens. The immunoreactions intensity was classified as weak, moderate and strong representing 2, 22 and 24%, respectively. According to multivariate analysis, EGFR expression was not significantly associated with age, sex, tumor site, stage, grade and type of tumor in cases. These results suggest that the assessment of EGFR expression in colorectal cancer by conventional immunohistochemistry has not proven its predictive value and can not be useful to predict about outcome of patients.


Mombeinipour M.,Payam Noor University of Mashhad
Archives of Iranian medicine | Year: 2013

Agiogenesis is the development of new blood vessels from pre-existing vasculatures. Although essential in the physiological process, it becomes pathological in various diseases including cancer. Preventing the formation of new blood vessels causes reductions in tumor size and metastasis. This study has been undertaken to elucidate the anti-angiogenesis effects of ICD-85 (derived peptides from venom). We evaluated the ICD-85 anti-angiogenesis activity by the in vivo CAM assay and in vitro tube formation assay of human umbilical vein endothelial cells (HUVECs). The anti-proliferative activity of ICD-85 was also determined through MTT assay on HUVECs. Results of this study revealed the anti-proliferative activity of ICD-85 on the HUVEC cell line with an IC50 of 12 μg/mL. The in vivo CAM assay also clearly showed the prevention of new vascular formation when the chick embryos were exposed to 0.15 μg/disc of ICD-85. In vitro tube formation assay of HUVECs also showed the complete prevention of capillary tube formation on 18 μg/mL. Based on the results obtained in this study, ICD-85 has anti-angiogenesis activity as shown by the prevention of capillary tube formation and the CAM assay.


PubMed | Payam Noor University of Mashhad
Type: Journal Article | Journal: Archives of Iranian medicine | Year: 2013

Agiogenesis is the development of new blood vessels from pre-existing vasculatures. Although essential in the physiological process, it becomes pathological in various diseases including cancer. Preventing the formation of new blood vessels causes reductions in tumor size and metastasis. This study has been undertaken to elucidate the anti-angiogenesis effects of ICD-85 (derived peptides from venom).We evaluated the ICD-85 anti-angiogenesis activity by the in vivo CAM assay and in vitro tube formation assay of human umbilical vein endothelial cells (HUVECs). The anti-proliferative activity of ICD-85 was also determined through MTT assay on HUVECs.Results of this study revealed the anti-proliferative activity of ICD-85 on the HUVEC cell line with an IC50 of 12 g/mL. The in vivo CAM assay also clearly showed the prevention of new vascular formation when the chick embryos were exposed to 0.15 g/disc of ICD-85. In vitro tube formation assay of HUVECs also showed the complete prevention of capillary tube formation on 18 g/mL.Based on the results obtained in this study, ICD-85 has anti-angiogenesis activity as shown by the prevention of capillary tube formation and the CAM assay.


PubMed | Payam Noor University of Mashhad
Type: Journal Article | Journal: Drug metabolism and drug interactions | Year: 2013

Methocarbamol is a skeletal muscle relaxant and is widely used to relieve pain in muscles. Many drugs may have interactions with each other when used at the same time. Yeast sucrase is taken as a drug by patients with congenital sucrase-isomaltase deficiency (CSID).In this study, the interaction between methocarbamol and yeast sucrase was investigated.Our results showed that methocarbamol can inhibit sucrase activity and reduce the maximum reaction velocity (Vmax) of the enzyme by a non-competitive pattern. Measurement of IC50 and Ki of the drug revealed that methocarbamol did not bind the enzyme with high affinity. Fluorescence measurement showed that the drug binds to free enzyme and enzyme-substrate complexes that were accompanied by structural changes on the enzyme. Guaifenesin, which has a similar structure to methocarbamol, does not affect the activity of sucrase.Methocarbamol inhibits sucrase activity and its carbamate group plays a main role in the binding of drug to sucrase.

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