Background: Colorectal cancer, the third most common type of cancer is a major cause of mortality... more Background: Colorectal cancer, the third most common type of cancer is a major cause of mortality worldwide. If colorectal cancer is detected at the early stages, the 5-year survival rate is 90%. MEIS1 homeobox gene is implicated in numerous solid tumors and hematological malignancies. Therefore, the present study aims to investigate the methylation status of the MEIS1 gene in colorectal cancer. Methods: We used real-time quantitative methylation-specific PCR to detect MEIS1 promoter methylation in 42 colorectal cancer tissues and 42 normal colorectal tissues. Results: Methylation was observed only in the positive control samples - CG Genome Universal Unmethylated DNA and CG Genome Universal Methylated DNA. There was no change observed in MEIS1 promoter methylation status in 42 patients. Conclusion: The results of the current study indicated that the MEIS1 gene promoter was not methylated in the cases. Gene expression study confirmed the unmethylated status of the MEIS1 gene in the ...
Objective(s): Human Cytomegalovirus (HCMV) remains a major morbidity and mortality cause in immun... more Objective(s): Human Cytomegalovirus (HCMV) remains a major morbidity and mortality cause in immuno suppressed patients. Therefore, significant effort has been made towards the development of a vaccine. In this study, the expression of the pp65 and gB fusion peptides and Fc domain of mouse IgG2a as a novel delivery system for selective uptake of antigens by antigen-presenting cells (APCs) in Pichia pastoris yeast system were studied. Materials and Method: In this study, four immune dominant sequences in pp65 protein and 3 immuno dominant sequences in gB protein were selected according to literature review. Peptide linker-GGGGS-was used for construction of fusion peptide. This fusion peptide was cloned in the pPICZαA expression vector and transfected into P. pastoris host cells. Results: Dot blot and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) techniques showed that a high level of pp65-gB-Fc fusion peptide was expressed. Conclusion: This CMV pp65-gB-Fc fusion...
Tissue factor (TF) is the core reagent in the prothrombin time (PT) assay. In this study, express... more Tissue factor (TF) is the core reagent in the prothrombin time (PT) assay. In this study, expression and α-factor mediated secretion of three forms of tissue factor (full-length TF (Full-TF), extracellular plus transmembrane domain (TED-TF), and only extracellular domain (ED-TF) were investigated in Pichia pastoris. The amino acid sequence of TF was obtained from the UniProt database, back-translated and codon-optimized for expression in Pichia pastoris. The Full-TF sequence was synthesized but the ED-TF, TED-TF coding fragments were extracted from the Full-TF by PCR. All the coding sequences were cloned into pPICZαA vector in-frame with the α-factor; and electroporated into KM71H. The culture supernatants and the cell lysates were analyzed using SDS-PAGE, dot-blotting, and Western-blotting for expression of TF. The Full-TF and TED-TF expression vector pPICZαA were successfully inserted into the KM71H, but the product was not detected in the SDS-PAGE analysis of the culture supernatant. However, ED-TF expression and secretion was verified by SDS-PAGE, dot blotting, and Western blotting. It seems that the TM domain in the Full-TF and TED-TF have an important role in impairing α-factor-mediated secretion of TF. Therefore, further investigation is necessary to overcome challenges of expressing Full-TF as a heterologous protein in P. pastoris.
Mutant forms FMS-like tyrosine kinase-3 (FLT3), are reported in 25% of childhood acute lymphoid l... more Mutant forms FMS-like tyrosine kinase-3 (FLT3), are reported in 25% of childhood acute lymphoid leukemia (ALL) and 30% of acute myeloid leukemia (AML) patients. In this study, drug response, growth promoting, and protein trafficking of FLT3 wild-type was compared with two active mutants (Internal Tandem Duplication (ITD)) and D835Y. FLT3 was expressed on factor-dependent cells (FDC-P1) using retroviral transduction. The inhibitory effects of CEP701, imatinib, dasatinib, PKC412 and sunitinib were studied on cell proliferation and FLT3 tyrosine phosphorylation. Total expression and proportion of intracellular and surface FLT3 was also determined. FDC-P1 cells became factor-independent after expression of human FLT3 mutants (ITD and D835Y). FDC-P1 cells expressing FLT3-ITD grow 3 to 4 times faster than those expressing FLT3-D835Y. FD-FLT3-ITD cells were three times more resistant to sunitinib than the FD-FLT3-WT cells. The Geo means for surface FLT3 expression in FD-FLT3-ITD and -D835Y...
Introduction: Galectin-3 is known as a biomarker in patients with heart failure. This protein par... more Introduction: Galectin-3 is known as a biomarker in patients with heart failure. This protein participates in different mechanisms involved in atherosclerosis including inflammation and plaque formation. This study was conducted to investigate whether this factor could be a predictive biomarker for the severity of atherosclerosis. Materials and Methods: The study group consisted of 80 patients with coronary atherosclerosis referred to the Department of Cardiac Surgery of Ghaem Hospital, affiliated to Mashhad University of Medical Sciences, Mashhad, Iran. The serum level of galectin-3 was measured using a commercial enzyme-linked immunosorbent assay kit. The severity of coronary artery disease (CAD), evaluated by the serum levels of galectin-3, was expressed as the number of involved vessels. Results: Galectin-3 concentration was directly correlated with the number of involved vessels.The serum level of galectin-3 was significantly higher in patients with four involved vessels (20.76...
Canadian journal of physiology and pharmacology, 2020
Diabetic cardiomyopathy (DC) is associated with impaired endoplasmic reticulum (ER) function, dev... more Diabetic cardiomyopathy (DC) is associated with impaired endoplasmic reticulum (ER) function, development of ER stress, and induction of cardiac cell apoptosis. Preventive effects of BiP inducer X (BIX) were investigated against DC characteristic changes in a type 2 diabetes rat model. To establish diabetes, a high-fat diet and a single dose of streptozotocin were administered. Then, animals were assigned into following groups: control, BIX, diabetic animals monitored for one, two, and three weeks. Diabetic rats treated with BIX for one, two, and three weeks. Expressions of various ER stress and apoptotic markers were assessed by immunoblotting method. CHOP gene expression was assessed by Real-time PCR. Tissue expression of BiP was evaluated by immunohistochemistry method. Hematoxylin and eosin and Masson's trichrome staining were performed to assess histological changes in the left ventricle. Cardiac cell apoptosis was examined using TUNEL assay. BIX administration suppressed t...
Objective Early diagnosis is has a crucial role in both prevention and treatment of asphyxia-rela... more Objective Early diagnosis is has a crucial role in both prevention and treatment of asphyxia-related complications. The current study aimed to evaluate the prognostic value of interleukin-6 (IL-6) and hypoxic-ischemic encephalopathy grade in the prediction of mortality and the developmental status of neonates affected by prenatal asphyxia. Materials & Methods This cohort study was conducted on 38 term asphyxiated infants at Ghaem hospital, Mashhad, Iran, from 2013 to 2017. The HIE grade and serum IL-6 levels were determined at the time of birth. The developmental status was evaluated using the Denver II test at the end of the two-year follow-up. Results HIE grade 3 resulted in 83% mortality rate and developmental delay among all survivors. The mean IL-6 level was 2.7 ng/ml in the control group (not affected HIE), which increased up to 29, 175, and 136 ng/ml in those with HIE grades of 1, 2, and 3, respectively. According to the ROC curve analysis, the cut-off level of 24 pg/ml could...
Golestan Rheumatology Research Center, Golestan University of Medical Sciences, Gorgan, Iran Depa... more Golestan Rheumatology Research Center, Golestan University of Medical Sciences, Gorgan, Iran Department of New Sciences and Technology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Pharmacological Research Center of Medicinal Plants, Mashhad University of Medical Sciences, Mashhad, Iran Non‐Communicable Diseases Research Center, Jahrom University of Medical Sciences, Jahrom, Iran Internal Medicine Department, Ghaem Hospital, Mashhad University of Medical Sciences, Mashhad, Iran Next Generation Genetic Clinic, Mashhad, Iran Department of Medical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Department of Molecular Biology and Biotechnology, Krebs and Sheffield Institute of Nucleic Acids, University of Sheffield, Sheffield, UK Infectious Diseases Research Center, Golestan University of Medical Sciences, Gorgan, Iran Division of Neurocognitive Sciences, Psychiatry and Behavioral Sciences Research Center, Mashhad U...
Objective(s): Mutant forms FMS-like tyrosine kinase-3 (FLT3), are reported in 25% of childhood ac... more Objective(s): Mutant forms FMS-like tyrosine kinase-3 (FLT3), are reported in 25% of childhood acute lymphoid leukemia (ALL) and 30% of acute myeloid leukemia (AML) patients. In this study, drug response, growth promoting, and protein trafficking of FLT3 wild-type was compared with two active mutants (Internal Tandem Duplication (ITD)) and D835Y. Materials and Methods: FLT3 was expressed on factor-dependent cells (FDC-P1) using retroviral transduction. The inhibitory effects of CEP701, imatinib, dasatinib, PKC412 and sunitinib were studied on cell proliferation and FLT3 tyrosine phosphorylation. Total expression and proportion of intracellular and surface FLT3 was also determined. Results: FDC-P1 cells became factor-independent after expression of human FLT3 mutants (ITD and D835Y). FDC-P1 cells expressing FLT3-ITD grow 3 to 4 times faster than those expressing FLT3-D835Y. FD-FLT3-ITD cells were three times more resistant to sunitinib than the FD-FLT3-WT cells. The Geo means for sur...
Objective(s): Human Cytomegalovirus (HCMV) remains a major morbidity and mortality cause in immun... more Objective(s): Human Cytomegalovirus (HCMV) remains a major morbidity and mortality cause in immuno suppressed patients. Therefore, significant effort has been made towards the development of a vaccine. In this study, the expression of the pp65 and gB fusion peptides and Fc domain of mouse IgG2a as a novel delivery system for selective uptake of antigens by antigen-presenting cells (APCs) in Pichia pastoris yeast system were studied. Materials and Method: In this study, four immune dominant sequences in pp65 protein and 3 immuno dominant sequences in gB protein were selected according to literature review. Peptide linker -GGGGS- was used for construction of fusion peptide. This fusion peptide was cloned in the pPICZαA expression vector and transfected into P. pastoris host cells. Results: Dot blot and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) techniques showed that a high level of pp65-gB-Fc fusion peptide was expressed. Conclusion: This CMV pp65-gB-Fc fusi...
Background and objective: β-galactosidase enzymes hydrolyze lactose into glucose and galactose fo... more Background and objective: β-galactosidase enzymes hydrolyze lactose into glucose and galactose for production of lactose free dairy products. However, different ions and fat content in milk may act as the inhibitor or activator for β-galactosidase enzymes. A cold-active β-galactosidase enzyme (BGalP), originally from Planococcus sp. L4 , was previously expressed in Pichia pastoris to perform lactose hydrolysis in the refrigerated milk. In this study, the effects of milks major ingredients were evaluated on the enzymatic kinetics to confirm its capacity for hydrolyzing milk lactose. Material and methods: The activity was determined in different concentrations of NaCl, KCl, MgCl 2, and CaCl 2 as well as in the milk with low, medium or high-fat content. In these experiments ortho-Nitrophenyl β-galactoside was used as the substrate. Additionally, glucose was measured as the product after incubation of milk with BGalP enzyme for 24 h at room temperature. Results and conclusion: This stud...
Background: The MET receptor is a critical member of cancer-associated RTKs and plays an importan... more Background: The MET receptor is a critical member of cancer-associated RTKs and plays an important role in different biological activities, including differentiation, migration, and cell proliferation. Methods: In this study, novel MET inhibitors were introduced and applied on esophageal squamous carcinoma cell line KYSE-30, and the level of proliferation and migration, as well as the activated form of MET receptor protein were assessed in the examined cells. The human KYSE-30 cell line was cultured according to ATCC recommendations. The mRNA level of the MET gene was measured in the examined cell line using the quantitative RT-PCR assay. Cytotoxicity evaluation test was performed at different concentrations of heterocyclic anti-MET compounds (i.e. D1, D2, D5, D6, D7, and D8). Finally, the capability of these compounds in MET receptor inhibition was evaluated using the migration assay and Western blot. All experiments were performed in triplicate and repeated three times with simila...
Immunotherapy through immune checkpoints blockade and its subsequent clinical application has rev... more Immunotherapy through immune checkpoints blockade and its subsequent clinical application has revolutionized the treatment of a spectrum of solid tumors. Blockade of Programmed cell death protein-1 and its ligand has shown promising results in clinical studies. The clinical trials that enrolled patients with different hematopoietic malignancies including non-Hodgkin lymphoma, Hodgkin lymphoma, and acute myeloid leukemia (AML) showed that anti-PD-1 agents could have potential therapeutic effects in the patients. Adult T-cell leukemia/lymphoma (ATLL) is a non-Hodgkin T-cell Lymphoma that is developed in a minority of HTLV-1-infected individuals after a long latency period. The inhibition of PD-1 as a treatment option is currently being investigated in ATLL patients. In this review, we present a summary of the biology of the PD-1/PD-L1 pathway, the evidence in the literature to support anti-PD-1/PDL-1 application in the treatment of different lymphoid, myeloid, and virus-related hematological malignancies, and controversies related to PD-1/PD-L1 blocking in the management of ATLL patients.
Tachykinins such as Substance P (SP) are a group of neuropeptides that are involved in cancer dev... more Tachykinins such as Substance P (SP) are a group of neuropeptides that are involved in cancer development. Neurokinin-1 receptor (NK-1R) is the main tachykinin receptor mediating the effects of SP, which is overexpressed in human esophageal squamous cell carcinoma (ESCC) and other malignant tissues. However, the effects of SP/NK-1R system on the migration of esophageal cancer cells and angiogenesis is not clear yet. This study seeks to obtain data to address these research gaps. In order to assess the effects of the FDA-approved aprepitant drug, a commercially available NK-1R antagonist, on the viability of KYSE-30 ESCC cells, resazurin assay was performed. The influence of SP/NK-1R system on the migration potential of these cells was examined using scratch assay. The effects of this system on the expression levels of metastatic factors were also examined by RT-PCR and western blot analyses. The half-maximal inhibitory concentration (IC 50 ) value for KYSE-30 cells treated with aprepitant found to be 29.88 μM. Treatment with SP significantly promoted KYSE-30 esophageal cancer cell migration, and aprepitant blocked this effect. In addition, SP significantly induced the expression of matrix metalloproteinase-2 (MMP-2), MMP-9, vascular endothelial growth factor-A (VEGF-A), and VEGF receptor1 (VEGFR1) in the cells, whereas aprepitant inhibited the up-regulation effects caused by SP. SP plays important roles in the development of human esophageal squamous cell carcinoma by promoting cancer cell invasion and enhancing the expression of factors involved in cellular migration and angiogenesis, which can be blocked by the NK-1R antagonist, aprepitant.
Introoduction: Inhibition of the reverse transcriptase (RT) enzyme of the human immunodeficiency ... more Introoduction: Inhibition of the reverse transcriptase (RT) enzyme of the human immunodeficiency virus (HIV) by low molecular weight inhibitors is still an active area of research. Here, protein-ligand interactions and possible binding modes of novel compounds with the HIV-1 RT binding pocket (the wild-type as well as Y181C and K103N mutants) were obtained and discussed. Methods: A molecular fragment-based approach using FDA-approved drugs were followed to design novel chemical derivatives using delavirdine, efavirenz, etravirine and rilpivirine as the scaffolds. The drug-likeliness of the derivatives was evaluated using Swiss-ADME. The parent molecule and derivatives were then docked into the binding pocket of related crystal structures (PDB ID: 4G1Q, 1IKW, 1KLM and 3MEC). Genetic Optimization for Ligand Docking (GOLD) Suite 5.2.2 software was used for docking and the results analyzed in the Discovery Studio Visualizer 4. A derivative was chosen for further analysis, if it passed d...
Crocetin, the major carotenoid in saffron, exhibits potent anticancer effects. However, the antil... more Crocetin, the major carotenoid in saffron, exhibits potent anticancer effects. However, the antileukemic effects of crocetin are still unclear, especially in primary acute promyelocytic leukemia (APL) cells. In the current study, the potential antipromyelocytic leukemia activity of crocetin and the underlying molecular mechanisms were investigated. Crocetin (100 µM), like standard anti‐APL drugs, all‐trans retinoic acid (ATRA, 10 µM) and As2O 3 (arsenic trioxide, 50 µM), significantly inhibited proliferation and induced apoptosis in primary APL cells, as well as NB4 and HL60 cells. The effect was associated with the decreased expressions of prosurvival genes Akt and BCL2, the multidrug resistance (MDR) proteins, ABCB1 and ABCC1 and the inhibition of tyrosyl‐DNA phosphodiesterase 1 (TDP1), while the expressions of proapoptotic genes CASP3, CASP9, and BAX/BCL2 ratio were significantly increased. In contrast, crocetin at relatively low concentration (10 µM), like ATRA (1 µM) and As 2O 3 (0.5 µM), induced differentiation of leukemic cells toward granulocytic pattern, and increased the number of differentiated cells expressing CD11b and CD14, while the number of the immature cells expressing CD34 or CD33 was decreased. Furthermore, crocetin suppressed the expression of clinical marker promyelocytic leukemia/retinoic acid receptor‐α ( PML/RARα) in NB4 and primary APL cells, and reduced the expression of histone deacetylase 1 ( HDAC1) in all leukemic cells. The results suggested that crocetin can be considered as a candidate for future preclinical and clinical trials of complementary APL treatment.
Objective Thrombin, platelets, and plasmin are three key factors involved in hemostasis and throm... more Objective Thrombin, platelets, and plasmin are three key factors involved in hemostasis and thrombolysis. Thrombolytic therapy with clinically approved drugs is often followed by recurrent thrombosis caused by thrombin-induced platelet aggregation from the clot debris. In order to minimize these problems, new constructs were designed for the expression of recombinant staphylokinase (rSAK) and also a fusion protein composed of staphylokinase, 20 amino acids containing 2 RGD followed by tsetse thrombin Inhibitor (SAK-2RGD-TTI) in Pichia pastoris . Result Modeling the tertiary structure of SAK-2RGD-TTI showed that the linker containing RGD and TTI did not interfere with proper folding of SAK. In laboratory testing, the purified SAK-2RGD-TTI (420 μg/mL) dissolved an average of 45% of the blood clot. The activity of the SAK-2RGD-TTI was also confirmed in various tests including human plasminogen activation assay, fibrin clot lysis assay, well diffusion method, activated partial thromboplastin time and platelet rich clot lysis assay. Conclusion Our findings suggest that SAK-2RGD-TTI has improved therapeutic properties preventing reocclussion. It further confirms that it is practicable to assemble and produce a hybrid multifunctional protein that targets hemostatic process at various stages.
Background: Colorectal cancer, the third most common type of cancer is a major cause of mortality... more Background: Colorectal cancer, the third most common type of cancer is a major cause of mortality worldwide. If colorectal cancer is detected at the early stages, the 5-year survival rate is 90%. MEIS1 homeobox gene is implicated in numerous solid tumors and hematological malignancies. Therefore, the present study aims to investigate the methylation status of the MEIS1 gene in colorectal cancer. Methods: We used real-time quantitative methylation-specific PCR to detect MEIS1 promoter methylation in 42 colorectal cancer tissues and 42 normal colorectal tissues. Results: Methylation was observed only in the positive control samples - CG Genome Universal Unmethylated DNA and CG Genome Universal Methylated DNA. There was no change observed in MEIS1 promoter methylation status in 42 patients. Conclusion: The results of the current study indicated that the MEIS1 gene promoter was not methylated in the cases. Gene expression study confirmed the unmethylated status of the MEIS1 gene in the ...
Objective(s): Human Cytomegalovirus (HCMV) remains a major morbidity and mortality cause in immun... more Objective(s): Human Cytomegalovirus (HCMV) remains a major morbidity and mortality cause in immuno suppressed patients. Therefore, significant effort has been made towards the development of a vaccine. In this study, the expression of the pp65 and gB fusion peptides and Fc domain of mouse IgG2a as a novel delivery system for selective uptake of antigens by antigen-presenting cells (APCs) in Pichia pastoris yeast system were studied. Materials and Method: In this study, four immune dominant sequences in pp65 protein and 3 immuno dominant sequences in gB protein were selected according to literature review. Peptide linker-GGGGS-was used for construction of fusion peptide. This fusion peptide was cloned in the pPICZαA expression vector and transfected into P. pastoris host cells. Results: Dot blot and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) techniques showed that a high level of pp65-gB-Fc fusion peptide was expressed. Conclusion: This CMV pp65-gB-Fc fusion...
Tissue factor (TF) is the core reagent in the prothrombin time (PT) assay. In this study, express... more Tissue factor (TF) is the core reagent in the prothrombin time (PT) assay. In this study, expression and α-factor mediated secretion of three forms of tissue factor (full-length TF (Full-TF), extracellular plus transmembrane domain (TED-TF), and only extracellular domain (ED-TF) were investigated in Pichia pastoris. The amino acid sequence of TF was obtained from the UniProt database, back-translated and codon-optimized for expression in Pichia pastoris. The Full-TF sequence was synthesized but the ED-TF, TED-TF coding fragments were extracted from the Full-TF by PCR. All the coding sequences were cloned into pPICZαA vector in-frame with the α-factor; and electroporated into KM71H. The culture supernatants and the cell lysates were analyzed using SDS-PAGE, dot-blotting, and Western-blotting for expression of TF. The Full-TF and TED-TF expression vector pPICZαA were successfully inserted into the KM71H, but the product was not detected in the SDS-PAGE analysis of the culture supernatant. However, ED-TF expression and secretion was verified by SDS-PAGE, dot blotting, and Western blotting. It seems that the TM domain in the Full-TF and TED-TF have an important role in impairing α-factor-mediated secretion of TF. Therefore, further investigation is necessary to overcome challenges of expressing Full-TF as a heterologous protein in P. pastoris.
Mutant forms FMS-like tyrosine kinase-3 (FLT3), are reported in 25% of childhood acute lymphoid l... more Mutant forms FMS-like tyrosine kinase-3 (FLT3), are reported in 25% of childhood acute lymphoid leukemia (ALL) and 30% of acute myeloid leukemia (AML) patients. In this study, drug response, growth promoting, and protein trafficking of FLT3 wild-type was compared with two active mutants (Internal Tandem Duplication (ITD)) and D835Y. FLT3 was expressed on factor-dependent cells (FDC-P1) using retroviral transduction. The inhibitory effects of CEP701, imatinib, dasatinib, PKC412 and sunitinib were studied on cell proliferation and FLT3 tyrosine phosphorylation. Total expression and proportion of intracellular and surface FLT3 was also determined. FDC-P1 cells became factor-independent after expression of human FLT3 mutants (ITD and D835Y). FDC-P1 cells expressing FLT3-ITD grow 3 to 4 times faster than those expressing FLT3-D835Y. FD-FLT3-ITD cells were three times more resistant to sunitinib than the FD-FLT3-WT cells. The Geo means for surface FLT3 expression in FD-FLT3-ITD and -D835Y...
Introduction: Galectin-3 is known as a biomarker in patients with heart failure. This protein par... more Introduction: Galectin-3 is known as a biomarker in patients with heart failure. This protein participates in different mechanisms involved in atherosclerosis including inflammation and plaque formation. This study was conducted to investigate whether this factor could be a predictive biomarker for the severity of atherosclerosis. Materials and Methods: The study group consisted of 80 patients with coronary atherosclerosis referred to the Department of Cardiac Surgery of Ghaem Hospital, affiliated to Mashhad University of Medical Sciences, Mashhad, Iran. The serum level of galectin-3 was measured using a commercial enzyme-linked immunosorbent assay kit. The severity of coronary artery disease (CAD), evaluated by the serum levels of galectin-3, was expressed as the number of involved vessels. Results: Galectin-3 concentration was directly correlated with the number of involved vessels.The serum level of galectin-3 was significantly higher in patients with four involved vessels (20.76...
Canadian journal of physiology and pharmacology, 2020
Diabetic cardiomyopathy (DC) is associated with impaired endoplasmic reticulum (ER) function, dev... more Diabetic cardiomyopathy (DC) is associated with impaired endoplasmic reticulum (ER) function, development of ER stress, and induction of cardiac cell apoptosis. Preventive effects of BiP inducer X (BIX) were investigated against DC characteristic changes in a type 2 diabetes rat model. To establish diabetes, a high-fat diet and a single dose of streptozotocin were administered. Then, animals were assigned into following groups: control, BIX, diabetic animals monitored for one, two, and three weeks. Diabetic rats treated with BIX for one, two, and three weeks. Expressions of various ER stress and apoptotic markers were assessed by immunoblotting method. CHOP gene expression was assessed by Real-time PCR. Tissue expression of BiP was evaluated by immunohistochemistry method. Hematoxylin and eosin and Masson's trichrome staining were performed to assess histological changes in the left ventricle. Cardiac cell apoptosis was examined using TUNEL assay. BIX administration suppressed t...
Objective Early diagnosis is has a crucial role in both prevention and treatment of asphyxia-rela... more Objective Early diagnosis is has a crucial role in both prevention and treatment of asphyxia-related complications. The current study aimed to evaluate the prognostic value of interleukin-6 (IL-6) and hypoxic-ischemic encephalopathy grade in the prediction of mortality and the developmental status of neonates affected by prenatal asphyxia. Materials & Methods This cohort study was conducted on 38 term asphyxiated infants at Ghaem hospital, Mashhad, Iran, from 2013 to 2017. The HIE grade and serum IL-6 levels were determined at the time of birth. The developmental status was evaluated using the Denver II test at the end of the two-year follow-up. Results HIE grade 3 resulted in 83% mortality rate and developmental delay among all survivors. The mean IL-6 level was 2.7 ng/ml in the control group (not affected HIE), which increased up to 29, 175, and 136 ng/ml in those with HIE grades of 1, 2, and 3, respectively. According to the ROC curve analysis, the cut-off level of 24 pg/ml could...
Golestan Rheumatology Research Center, Golestan University of Medical Sciences, Gorgan, Iran Depa... more Golestan Rheumatology Research Center, Golestan University of Medical Sciences, Gorgan, Iran Department of New Sciences and Technology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Pharmacological Research Center of Medicinal Plants, Mashhad University of Medical Sciences, Mashhad, Iran Non‐Communicable Diseases Research Center, Jahrom University of Medical Sciences, Jahrom, Iran Internal Medicine Department, Ghaem Hospital, Mashhad University of Medical Sciences, Mashhad, Iran Next Generation Genetic Clinic, Mashhad, Iran Department of Medical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Department of Molecular Biology and Biotechnology, Krebs and Sheffield Institute of Nucleic Acids, University of Sheffield, Sheffield, UK Infectious Diseases Research Center, Golestan University of Medical Sciences, Gorgan, Iran Division of Neurocognitive Sciences, Psychiatry and Behavioral Sciences Research Center, Mashhad U...
Objective(s): Mutant forms FMS-like tyrosine kinase-3 (FLT3), are reported in 25% of childhood ac... more Objective(s): Mutant forms FMS-like tyrosine kinase-3 (FLT3), are reported in 25% of childhood acute lymphoid leukemia (ALL) and 30% of acute myeloid leukemia (AML) patients. In this study, drug response, growth promoting, and protein trafficking of FLT3 wild-type was compared with two active mutants (Internal Tandem Duplication (ITD)) and D835Y. Materials and Methods: FLT3 was expressed on factor-dependent cells (FDC-P1) using retroviral transduction. The inhibitory effects of CEP701, imatinib, dasatinib, PKC412 and sunitinib were studied on cell proliferation and FLT3 tyrosine phosphorylation. Total expression and proportion of intracellular and surface FLT3 was also determined. Results: FDC-P1 cells became factor-independent after expression of human FLT3 mutants (ITD and D835Y). FDC-P1 cells expressing FLT3-ITD grow 3 to 4 times faster than those expressing FLT3-D835Y. FD-FLT3-ITD cells were three times more resistant to sunitinib than the FD-FLT3-WT cells. The Geo means for sur...
Objective(s): Human Cytomegalovirus (HCMV) remains a major morbidity and mortality cause in immun... more Objective(s): Human Cytomegalovirus (HCMV) remains a major morbidity and mortality cause in immuno suppressed patients. Therefore, significant effort has been made towards the development of a vaccine. In this study, the expression of the pp65 and gB fusion peptides and Fc domain of mouse IgG2a as a novel delivery system for selective uptake of antigens by antigen-presenting cells (APCs) in Pichia pastoris yeast system were studied. Materials and Method: In this study, four immune dominant sequences in pp65 protein and 3 immuno dominant sequences in gB protein were selected according to literature review. Peptide linker -GGGGS- was used for construction of fusion peptide. This fusion peptide was cloned in the pPICZαA expression vector and transfected into P. pastoris host cells. Results: Dot blot and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) techniques showed that a high level of pp65-gB-Fc fusion peptide was expressed. Conclusion: This CMV pp65-gB-Fc fusi...
Background and objective: β-galactosidase enzymes hydrolyze lactose into glucose and galactose fo... more Background and objective: β-galactosidase enzymes hydrolyze lactose into glucose and galactose for production of lactose free dairy products. However, different ions and fat content in milk may act as the inhibitor or activator for β-galactosidase enzymes. A cold-active β-galactosidase enzyme (BGalP), originally from Planococcus sp. L4 , was previously expressed in Pichia pastoris to perform lactose hydrolysis in the refrigerated milk. In this study, the effects of milks major ingredients were evaluated on the enzymatic kinetics to confirm its capacity for hydrolyzing milk lactose. Material and methods: The activity was determined in different concentrations of NaCl, KCl, MgCl 2, and CaCl 2 as well as in the milk with low, medium or high-fat content. In these experiments ortho-Nitrophenyl β-galactoside was used as the substrate. Additionally, glucose was measured as the product after incubation of milk with BGalP enzyme for 24 h at room temperature. Results and conclusion: This stud...
Background: The MET receptor is a critical member of cancer-associated RTKs and plays an importan... more Background: The MET receptor is a critical member of cancer-associated RTKs and plays an important role in different biological activities, including differentiation, migration, and cell proliferation. Methods: In this study, novel MET inhibitors were introduced and applied on esophageal squamous carcinoma cell line KYSE-30, and the level of proliferation and migration, as well as the activated form of MET receptor protein were assessed in the examined cells. The human KYSE-30 cell line was cultured according to ATCC recommendations. The mRNA level of the MET gene was measured in the examined cell line using the quantitative RT-PCR assay. Cytotoxicity evaluation test was performed at different concentrations of heterocyclic anti-MET compounds (i.e. D1, D2, D5, D6, D7, and D8). Finally, the capability of these compounds in MET receptor inhibition was evaluated using the migration assay and Western blot. All experiments were performed in triplicate and repeated three times with simila...
Immunotherapy through immune checkpoints blockade and its subsequent clinical application has rev... more Immunotherapy through immune checkpoints blockade and its subsequent clinical application has revolutionized the treatment of a spectrum of solid tumors. Blockade of Programmed cell death protein-1 and its ligand has shown promising results in clinical studies. The clinical trials that enrolled patients with different hematopoietic malignancies including non-Hodgkin lymphoma, Hodgkin lymphoma, and acute myeloid leukemia (AML) showed that anti-PD-1 agents could have potential therapeutic effects in the patients. Adult T-cell leukemia/lymphoma (ATLL) is a non-Hodgkin T-cell Lymphoma that is developed in a minority of HTLV-1-infected individuals after a long latency period. The inhibition of PD-1 as a treatment option is currently being investigated in ATLL patients. In this review, we present a summary of the biology of the PD-1/PD-L1 pathway, the evidence in the literature to support anti-PD-1/PDL-1 application in the treatment of different lymphoid, myeloid, and virus-related hematological malignancies, and controversies related to PD-1/PD-L1 blocking in the management of ATLL patients.
Tachykinins such as Substance P (SP) are a group of neuropeptides that are involved in cancer dev... more Tachykinins such as Substance P (SP) are a group of neuropeptides that are involved in cancer development. Neurokinin-1 receptor (NK-1R) is the main tachykinin receptor mediating the effects of SP, which is overexpressed in human esophageal squamous cell carcinoma (ESCC) and other malignant tissues. However, the effects of SP/NK-1R system on the migration of esophageal cancer cells and angiogenesis is not clear yet. This study seeks to obtain data to address these research gaps. In order to assess the effects of the FDA-approved aprepitant drug, a commercially available NK-1R antagonist, on the viability of KYSE-30 ESCC cells, resazurin assay was performed. The influence of SP/NK-1R system on the migration potential of these cells was examined using scratch assay. The effects of this system on the expression levels of metastatic factors were also examined by RT-PCR and western blot analyses. The half-maximal inhibitory concentration (IC 50 ) value for KYSE-30 cells treated with aprepitant found to be 29.88 μM. Treatment with SP significantly promoted KYSE-30 esophageal cancer cell migration, and aprepitant blocked this effect. In addition, SP significantly induced the expression of matrix metalloproteinase-2 (MMP-2), MMP-9, vascular endothelial growth factor-A (VEGF-A), and VEGF receptor1 (VEGFR1) in the cells, whereas aprepitant inhibited the up-regulation effects caused by SP. SP plays important roles in the development of human esophageal squamous cell carcinoma by promoting cancer cell invasion and enhancing the expression of factors involved in cellular migration and angiogenesis, which can be blocked by the NK-1R antagonist, aprepitant.
Introoduction: Inhibition of the reverse transcriptase (RT) enzyme of the human immunodeficiency ... more Introoduction: Inhibition of the reverse transcriptase (RT) enzyme of the human immunodeficiency virus (HIV) by low molecular weight inhibitors is still an active area of research. Here, protein-ligand interactions and possible binding modes of novel compounds with the HIV-1 RT binding pocket (the wild-type as well as Y181C and K103N mutants) were obtained and discussed. Methods: A molecular fragment-based approach using FDA-approved drugs were followed to design novel chemical derivatives using delavirdine, efavirenz, etravirine and rilpivirine as the scaffolds. The drug-likeliness of the derivatives was evaluated using Swiss-ADME. The parent molecule and derivatives were then docked into the binding pocket of related crystal structures (PDB ID: 4G1Q, 1IKW, 1KLM and 3MEC). Genetic Optimization for Ligand Docking (GOLD) Suite 5.2.2 software was used for docking and the results analyzed in the Discovery Studio Visualizer 4. A derivative was chosen for further analysis, if it passed d...
Crocetin, the major carotenoid in saffron, exhibits potent anticancer effects. However, the antil... more Crocetin, the major carotenoid in saffron, exhibits potent anticancer effects. However, the antileukemic effects of crocetin are still unclear, especially in primary acute promyelocytic leukemia (APL) cells. In the current study, the potential antipromyelocytic leukemia activity of crocetin and the underlying molecular mechanisms were investigated. Crocetin (100 µM), like standard anti‐APL drugs, all‐trans retinoic acid (ATRA, 10 µM) and As2O 3 (arsenic trioxide, 50 µM), significantly inhibited proliferation and induced apoptosis in primary APL cells, as well as NB4 and HL60 cells. The effect was associated with the decreased expressions of prosurvival genes Akt and BCL2, the multidrug resistance (MDR) proteins, ABCB1 and ABCC1 and the inhibition of tyrosyl‐DNA phosphodiesterase 1 (TDP1), while the expressions of proapoptotic genes CASP3, CASP9, and BAX/BCL2 ratio were significantly increased. In contrast, crocetin at relatively low concentration (10 µM), like ATRA (1 µM) and As 2O 3 (0.5 µM), induced differentiation of leukemic cells toward granulocytic pattern, and increased the number of differentiated cells expressing CD11b and CD14, while the number of the immature cells expressing CD34 or CD33 was decreased. Furthermore, crocetin suppressed the expression of clinical marker promyelocytic leukemia/retinoic acid receptor‐α ( PML/RARα) in NB4 and primary APL cells, and reduced the expression of histone deacetylase 1 ( HDAC1) in all leukemic cells. The results suggested that crocetin can be considered as a candidate for future preclinical and clinical trials of complementary APL treatment.
Objective Thrombin, platelets, and plasmin are three key factors involved in hemostasis and throm... more Objective Thrombin, platelets, and plasmin are three key factors involved in hemostasis and thrombolysis. Thrombolytic therapy with clinically approved drugs is often followed by recurrent thrombosis caused by thrombin-induced platelet aggregation from the clot debris. In order to minimize these problems, new constructs were designed for the expression of recombinant staphylokinase (rSAK) and also a fusion protein composed of staphylokinase, 20 amino acids containing 2 RGD followed by tsetse thrombin Inhibitor (SAK-2RGD-TTI) in Pichia pastoris . Result Modeling the tertiary structure of SAK-2RGD-TTI showed that the linker containing RGD and TTI did not interfere with proper folding of SAK. In laboratory testing, the purified SAK-2RGD-TTI (420 μg/mL) dissolved an average of 45% of the blood clot. The activity of the SAK-2RGD-TTI was also confirmed in various tests including human plasminogen activation assay, fibrin clot lysis assay, well diffusion method, activated partial thromboplastin time and platelet rich clot lysis assay. Conclusion Our findings suggest that SAK-2RGD-TTI has improved therapeutic properties preventing reocclussion. It further confirms that it is practicable to assemble and produce a hybrid multifunctional protein that targets hemostatic process at various stages.
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Papers by Baratali Mashkani