Comparative proteome analysis of human esophageal cancer and adjacent normal tissues

Objective(s): Ranking as the sixth commonest cancer, esophageal squamous cell carcinoma (ESCC) represents one of the leading causes of cancer death worldwide. One of the main reasons for the low survival of patients with esophageal cancer is its late diagnosis. Materials and Methods: We used proteomics approach to analyze ESCC tissues with the aim of a better understanding of the malignant mechanism and searching candidate protein biomarkers for early diagnosis of esophageal cancer. The differential protein expression between cancerous               and normal esophageal tissues was investigated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Then proteins were identified by matrix-assisted laser desorption/ ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS) and MASCOT web based search engine. Results:We reported 4 differentially expressed proteins involved in the pathological process of esophageal cancer, such as annexinA1 (ANXA1), peroxiredoxin-2 (PRDX2), transgelin (TAGLN) andactin-aortic smooth muscle (ACTA2). Conclusion: In this report we have introduced new potential biomarker (ACTA2). Moreover, our data confirmed some already known markers for EC in our region

Aptamer-Conjugated PLGA Nanoparticles for Delivery and Imaging of Cancer Therapeutic Drugs

Most problems associated with chemotherapeutic agents involve non-specific cytotoxicity, low intratumoral accumulation and drug resistance. Targeted drug delivery systems (TDDS) based on nanoparticles (NPs) are a new strategy for better therapeutic efficiency, along with reduction of side effects commonly seen with cancer drugs. Poly (lactic-co-glycolic acid) (PLGA), as one of the furthest developed synthetic polymer, has gained significant attention because of excellent properties—including biodegradability and biocompatibility, controlled release of drug, protection of drug or gene from decomposition and ability to modify surface with targeting agents for both cancer diagnosis and therapy. Aptamers are single-stranded RNA or DNA that can fold through intramolecular interactions into specific three-dimensional structures to selectively and exclusively bind with interested biomarkers. In this review, we explain the latest developments regarding the application of aptamer-decorated PLGA NPs in delivery of therapeutic agents or cancer-related genes into cancer cells. Additionally, we discuss the most recent efforts in the field of aptamer-grafted PLGA-based NPs as theranostics and stimuli-responsive agents

Evaluation of Neuroprotective Effect of Althaea Officinalis Flower Aqueous and Methanolic Extracts against H2O2-Induced Oxidative Stress in PC12 Cells: Protective activity of Althaea Officinalis against oxidative stress

This study was conducted to evaluate the possible antioxidant activity and neuroprotective effects of aqueous and methanolic extracts of Althaeaofficinalis flowers against H2O2-induced oxidative stress in PC12 cells.The antioxidant potential of extracts was evaluated by radical scavenging activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. For cytoprotective activity, the cells were pretreated with different concentrations (12.5, 25, 50, 100, 200and 400 μg/ml) of the extracts for 24h and then incubated with H2O2(480 μM) for 3 h. In co-treatment protocol, cells were simultaneously treated with H2O2 (480 μM) and the same concentrations of extracts, used in pretreatment protocol. Percentage of viability was measured using MTT assay. The aqueous and methanolic extracts did not show strong DPPH radical scavenging activity (IC50 value of 128 and 255 μg/ml respectively) in comparison with ascorbic acid (IC50 value of 6.1 μg/ml). The cytoprtection study revealed that neither the methanolic, northe aqueous extractsat tested concentrations could protect the cells against H2O2-induced cytotoxicity compared to H2O2 alone, in either co-treatment or pre-treatment experiments. Despite reporting the antioxidant activity of A. officinalis L. flowers, it seems that such a negligible cytoprotective activity may be related to some other factors. On the other hand, the presence of moderate antioxidant activity does not guarantee the protective activity against oxidative stress

Anti-Melanogenic Activity of Different Extracts from Aerial Parts of Nepeta glomerulosa on Murine Melanoma B16F10 Cells: Anti-Melanogenic Activity of Nepeta glomeruloas on Murine Melanoma B16F10 Cells

Nepeta glomerulosa Boiss. is a medicinal plant used in traditional medicine. The aim of this study was to evaluate the anti-melanogenesis inhibitory activity of Methanol (MeOH), n-hexane, acetate (EtAc), dichloromethane (CH2Cl2), n-butanol (BuOH), and H2O extracts isolated from N. glomerulosa in B16 melanoma cell line. The B16F10 cell line viability after treatment with increasing concentrations of different extracts of the plant (50-200 μg/ml) was measured by using MTT. The inhibitory effect on synthesis of melanin, mushroom tyrosinase activity, cellular tyrosinase, and effect on oxidative stress was determined by the colorimetric and fluorometric method. The data showed that at concentrations <200 μg/ml, all extracts did not show significant toxicity on melanoma cells. The amount of melanin synthesis by MeOH and CH2Cl2 extracts and mushroom tyrosinase activity by the MeOH extract declined in B16F10 cells. In addition to the capacity of MeOH, n-hexane and CH2Cl2 extracts in decreasing the amount of reactive oxygen species (ROS) in melanoma cells all extracts revealed the remarkable antioxidant activity. Our results showed the melanogenesis inhibitory and antioxidant effects of N. glomerulosa on B16F10 cells may recommend a novel agent in diminishing skin pigmentation and skin aging in cosmetic industry

Involvement of brain-derived neurotrophic factor (BDNF) on malathion induced depressive-like behavior in subacute exposure and protective effects of crocin

Objective(s): In this study the effect of crocin, a carotenoid isolated from saffron, on malathion (an organophosphate insecticide) induced depressive- like behavior in subacute exposure was investigated. Moreover the molecular mechanism of malathion induced depressive- like behavior and its decreasing effect on the level of brain derived neurotrophic factor (BDNF) in rat hippocampus and cerebral cortex were evaluated. Materials and Methods: Male Wistar rats were exposed to malathion (50 mg/kg/day, IP) alone or in combination with crocin (10, 20 and 40 mg/kg/day, IP), imipramine (20 mg/kg/day, IP) and vitamin E (200 mg/kg, three times a week, IP) respectively for 14 days. The forced swimming test (FST) was performed on days 1st, 7th and 14st. The level of malondealdehyde (MDA) and reduced glutathione (GSH) were measured in cerebral cortex and hippocampus of rats. The protein level of BDNF was evaluated using Western blot analysis. Results: Malathion (50 mg/kg, IP) increased immobility time in the FST, without affecting total locomotor activity in open-field test. Malathion increased the malondealdehyde (MDA) and decreased the glutathione (GSH), whereas these effects were reversed by crocin and vitamin E. Malathion decreased plasma acetylcholinesterase  activity,  however  this effect was not reversed by crocin or vitamin E. Malathion reduced the protein level of BDNF in rat hippocampus. Imipramine and crocin  prevented the decreasing effect of malathion on BDNF. Conclusion: These results showed that crocin attenuates some neurochemical and behavioral effects induced by malathion. This neuroprotective effect of crocin may be in part due to its effect on BDNF

Screening and identification of SUMP-proteins in sub-acute treatment with diazinon

Objective(s):Small ubiquitin-like modifiers (SUMOs) are a family of ubiquitin-related, proteins that are involved in a wide variety of signaling pathways. SUMOylation, as a vital post translational modification, regulate protein function in manycellular processes. Diazinon (DZN), an organophosphate insecticide, causses oxidative stress and subsequently programmed cell death in different tissues. The aim of this study was to evaluate the role and pattern of SUMO modificationas a defense mechanism against stress oxidative, in the heart tissuesof the DZN treated rats. Materials and Methods: Diazinon (15 mg/kg/day), corn oil (control) were administered via gavageto male Wistar rats for four weeks. SUMO1 antibody was covalently crosslinked to protein A/G agarose. heart tissue lysate were added to agarosebeads,After isolation of target proteins(SUMO1- protein)SDS-PAGE gel electrophoresis was performed. Protein bands were identified using MALDI-TOF/TOF and MASCOT). Fold change of (DZN/Ctrl) separated proteins was evaluated using UVband software (UVITEC, UK). Results:Our result showed that subacute exposure to DZN increased SUMOylationoffour key proteins involved in the metabolic process including; Acyl-CoA dehydrogenase, creatine kinase, glyceraldehyde-3-phosphate dehydrogenase and ATP synthase, in the heart tissue of animals .A probability value of  less than 0.05 was considered significant (

CD44-specific short peptide A6 boosts cellular uptake and anticancer efficacy of PEGylated liposomal doxorubicin in vitro and in vivo

Abstract Although liposomes have improved patient safety and the pharmacokinetic profile of free drugs, their therapeutic efficacy has only shown marginal improvement. The incorporation of active-targeted ligands to enhance cellular uptake has shown promise in preclinical studies. However, no active-targeted liposomes have successfully translated into clinical use thus far. This study aimed to evaluate the targeting ability and antitumor efficiency of A6, a specific short peptide (KPSSPPEE) when incorporated into PEGylated liposomal doxorubicin (PLD). The results revealed significantly enhanced cellular uptake. The cytotoxicity of the formulations was determined by 3 h and 6 h incubation of formulations with cells, followed by 48 h incubation to evaluate the targeted ability of the formulations and the results indicated the higher cytotoxicity of A6-PLD (IC50 of 7.52 µg/mL after 6 h incubation) in the CD44 overexpressing C26 cell line compared to non-targeted PLD (IC50 of 15.02 µg/mL after 6 h incubation). However, CD44-negative NIH-3T3 cells exhibited similar uptake and in vitro cytotoxicity for both A6-PLD (IC50 of 38.05 µg/mL) and PLD (IC50 of 34.87 µg/mL). In animal studies, A6-PLD demonstrated significantly higher tumor localization of doxorubicin (Dox) (~ 8 and 15 µg Dox/g tumor for 24 and 48 after injection) compared to PLD (~ 6 and 8 µg Dox/g tumor for 24 and 48 after injection), resulting in effective inhibition of tumor growth. The median survival time (MST) for Dextrose 5% was 10, PLD was 14 and A6-PLD was 22 days. In conclusion, A6-PLD, a simple and effective targeted liposome formulation, exhibits high potential for clinical translation. Its improved targetability and antitumor efficacy make it a promising candidate for future clinical applications

Recognition and characterization of Erythropoietin binding-proteins in the brain of mice

Objective(s): Erythropoietin (EPO), is a 34KDa glycoprotein hormone, which belongs to type 1 cytokine superfamily. EPO involves in erythrocyte maturation through inhibition of apoptosis in erythroid cells. Besides its main function, protective effects of EPO in heart and brain tissues have been reported. EPO has a critical role in development, growth, and homeostasis of brain. Furthermore EPO has great potential in the recovery of different brain diseases which are still under studying. In this research, EPO binding pattern to brain proteins in animal model was studied. Materials and Methods:EPO antibody was covalently crosslinked to protein A/G agarose. in order to interact between EPO  and its target in brain,  about 5µg EPO added to brain homogenates(500ul of 1 mg/ml) and incubate at 4ο C for 30 min. brain tissue lysate were added to agarose beads, After isolation of target proteins(EPO - protein) both one and two-dimensional gel electrophoresis were performed. Proteins were identified utilizing MALDI-TOF/TOF and MASCOT software. Results: This research showed that EPO could physically interact with eightproteins including  Tubulin beta, Actin cytoplasmic 2, T-complex protein 1, TPR and ankyrin repeat-containing protein 1, Centromere-associated protein E, Kinesin-like protein KIF7, Growth arrest-specific protein 2 and  Pleckstrin homology-like domain family B member 2. Conclusion: Since EPO is a promising therapeutic drug for the treatment of neurological diseases, identified proteins may help us to have a better understanding about the mechanism of protective effects of EPO in the brain. Our data needs to be validated by complementary bioassays