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Aim: Detection of protein expression changes in human cystic echinococcosis sera by 2D gel electrophoresis
Background: Diagnosis and successful treatment of cystic echinococcosis [CE] is a major challenge, up to now. Identification of related expressed proteins using proteomics tools and bioinformatics analysis of patientsf sera have not been investigated, so far
methods: Sera from eight confirmed CE patients and three healthy controls were collected, tested by 2-DE for total protein separation of serum and analyzed using proteomics and bioinformatics methods. The gels were stained by Coomassie blue followed by scan imaging of the gels. The protein spots in each gel were analyzed using progenesis same spots software. Proteins names were obtained from TagIdent server
Results: A total of 263 protein spots with different expression were detected in both normal and diseased samples. Comparison between diseased and normal gels showed the expression of 45 up-regulated protein spots with fold.2 in diseased gel of which 10 were new proteins with statistical difference by normal gel [p-value<0.05]. On the other hand, the expression of 50 down-regulated protein spots were observed of which 11 proteins have been suppressed. Clustering of all detected sera proteins [263] using correlation analysis, divided the proteins into 2 clusters based on up-regulated and down-regulated expression of proteins. Clustering results were approved by principal component analysis [PCA]
Conclusion: Significant protein expression changes in human CE sera which is demonstrable by application of proteomics and bioinformatics analysis makes it an impressing tool for diagnosis of CE patients
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Introduction: During the last 3 decades, human is exposed to extremely low frequency electromagnetic fields [ELF-EMF] emitted by power lines and electronic devices. It is now well accepted that ELF-EMF are able to produce a variety of biological effects, although the molecular mechanism is unclear and controversial. Investigation of different intensities effects of 50 Hz ELF-EMF on cell morphology and protein expression is the aim of this study
Methods: SH-SY5Y human neuroblastoma cell line was exposed to 0.5 and 1 mT 50 Hz [ELF-EMF] for 3 hours. Proteomics techniques were used to determine the effects of these fields on protein expression. Bioinformatic and statistical analysis of proteomes were performed using Progensis SameSpots software
Results: Our results showed that exposure to ELF-EMF changes cell morphology and induces a dose-dependent decrease in the proliferation rate of the cells. The proteomic studies and bioinformatic analysis indicate that exposure to 50 Hz ELF-EMF leads to alteration of cell protein expression in both dose-dependent and intensity dependent manner, but the later is more pronounced
Conclusion: Our data suggests that increased intensity of ELF-EMF may be associated with more alteration in cell protein expression, as well as effect on cell morphology and proliferation
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Aim: The aim of this study was to provide a biomarker panel for esophageal, gastric and colorectal cancers. It can help introducing some diagnostic biomarkers for these diseases
Background: Gastrointestinal cancers [GICs] including esophageal, gastric and colorectal cancers are the most common cancers in the world which are usually diagnosed in the final stages and due to heterogeneity of these diseases, the treatments usually are not successful. For this reason, many studies have been conducted to discover predictive biomarkers
Methods: In the present study, 507 genes related to esophageal, gastric and colon cancers were extracted. The network was constructed by Cytoscape software [version 3.4.0]. Then a main component of the network was analyzed considering centrality parameters including degree, betweenness, closeness and stress. Three clusters of the protein network accompanied with their seed nodes were determined by MCODE application in Cytoscape software. Furthermore, Gene Ontology [GO] analysis of the key genes in combination to the seed nodes was performed
Results: The network of 17 common differential expressed genes in three esophageal, gastric and colon adenocarcinomas including 1730 nodes and 9188 edges were constructed. Eight crucial genes were determined. Three Clusters of the network were analyzed by GO analysis
Conclusion: The analyses of common genes of the three cancers showed that there are some common crucial genes including TP53, EGFR, MYC, AKT1, CDKN2A, CCND1 and HSP90AA1 which are tightly related to gastrointestinal cancers and can be predictive biomarkers for these cancers
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The aim of this investigation was conducted to proteomic analysis of plasma obtained from pregnant women who destined to develop late-onset preeclampsia without intrauterine growth restriction [IUGR] during 16[th] week of gestation. Plasma was obtained from primiparous women during 16[th] week of gestation. 2-DE proteomic analysis was done for plasma from 11 healthy pregnant women and 11 women who developed preeclampsia later. Using bioinformatic analysis with Progenesis SameSpots ver4.0 software and ANOVA test, expression of 2 spots were statistically different between two groups. In preeclamptic state, expression of both were decreased, one of these spots was vitamin D binding protein [p-value: 0.047], the other one will be discussed in another paper. According to results, we concluded that during 16[th] week of gestation, occurance of late-onset preeclampsia without IUGR is predictable. During this week, pathology of disease is present and may be the process of placental degeneration and impaired placentation are include in disease pathology
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Different treatment strategies of Alzheimers disease [AD] are being studied for treating or slowing the progression of AD. Many pharmaceutically important regulation systems operate through proteins as drug targets. Here, we investigate the drug target proteins in beta-amyloid [A beta] injected rat hippocampus treated with Lavandula angustifolia [LA] by proteomics techniques. The reported study showed that lavender extract [LE] improves the spatial performance in AD animal model by diminishing A beta production in histopathology of hippocampus, so in this study neuroprotective proteins expressed in A beta injected rats treated with LE were scrutinized. Rats were divided into three groups including normal, A beta injected, and A beta injected that was treated with LE. Protein expression profiles of hippocampus tissue were determined by two-dimensional electrophoresis [2DE] method and dysregulated proteins such as Snca, NF-L, Hspa5, Prdx2, Apoa1, and Atp5a1were identified by MALDI-TOF/TOF. KEGG pathway and gene ontology [GO] categories were used by searching DAVID Bioinformatics Resources. All detected protein spots were used to determine predicted interactions with other proteins in STRING online database. Different isoforms of important protein, Snca that exhibited neuroprotective effects by anti-apoptotic properties were expressed. NF-L involved in the maintenance of neuronal caliber. Hspa5 likewise Prdx2 displays as anti-apoptotic protein that Prdx2 also involved in the neurotrophic effects. Apoa1 has anti-inflammatory activity and Atp5a1, produces ATP from ADP. To sum up, these proteins as potential drug targets were expressed in hippocampus in response to effective components in LA may have therapeutic properties for the treatment of AD and other neurodegenerative diseases
Subject(s)
Animals, Laboratory , Alzheimer Disease/drug therapy , Drug Delivery Systems , Protein Transport , Models, Animal , RatsABSTRACT
Extremely low frequency electromagnetic fields [ELF-EMF] have been common in daily life all over the world. They have produced by power lines and electrical appliances, but higher levels of them have raised a lot of concerns about their carcinogenesis. Both epidemiological and laboratory studies have suggested that EMFs might increase cancer incidence, including acute childhood leukemia, brain and breast cancer. In the present study, SH-SY5Y human neuroblastoma cell line has exposed to 2mT, 50 Hz magnetic field for 3 h. Next, effect of this exposure on protein expression including over-expression or under-expression has assessed by proteomics. Bioinformatics and statistical analysis using progenesis same spot software on the obtained 2D electrophoresis has shown that expression of 189 proteins in exposed group has changed relative to control. Besides, PCA analysis has verified results of clustering, and has shown that protein data has clustered according to experimental conditions. The results of this study have shown that ELF-EMF changes cell morphology via altering protein expression, but more profound studies have needed to determine the kind of proteins altered
Subject(s)
Humans , Neuroblastoma , Cell Line , ProteomicsABSTRACT
Ethanol known as ethyl alcohol is being widely used around the world. Many serious diseases are related with its consumption. Alcohol posses many divers effects on human body including risk of cirrhosis and/or hepatocellular carcinoma. Therefore, analysis of this component is prominent. Fibroblast cells were cultured in various dosages of ethanol. The effective dosage was then investigated by proteomic methods. Separated proteins of fibroblast cells by Two-Dimensional Gel [2DG] Electrophoresis method based on pI and MW were analyzed based on spots alteration by Same Spot Software. Furthered analysis was carried out with vigorous statistical analysis based on significant folding changes and one-way ANOVA. About 372 protein spots were identified and among them 65 of them were having significant expression profile, which is evaluated as p <0.05. Therefore, ethanol can induce a great impact on protein profile of fibroblast. It is concluded that altering morphologic features and viability, as well as protein expression changes, confirm toxic properties of ethanol in human body
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Saprophitic Pseudomonas species are root-colonizing bacteria that can improve plant health. Efficient exploitation of these bacteria in agriculture requires knowledge of traits that enhance ecological performance in the rhizosphere. Some Pseudomonas fluorescens strains present biocontrol properties, protecting the roots of some plant species against plant pathogens. These bacteria induce systemic resistance in the host plant, so it can better resist attack by a true pathogen. The bacteria outcompete other [pathogenic] soil microbes, e.g., by siderophores, giving a competitive advantage at scavenging for iron. The bacteria produce compounds antagonistic to other soil microbes, such as phenazine - type antibiotics or hydrogen cyanide. In this study the changes in the protein profile of P. fluorescens strain UTPF68, involved in the multiple interactions between plant [tomato] and an antagonistic agent [Trichoderma atroviride strain P1] investigated. Two-dimensional electrophoresis was used to analyze separately collected proteins from each one, two or three partner interactions. The results about differential produced spots in Pseudomonas proteome in each collation, showed that 18 differential spots became visible as new, 16 spots were absent, 17 spots were up-regulated and 1 spot was down-regulated, when Tomato-Pseudomonas [TP] condition was compared with control Pseudomonas alone [P]. Also more than 84 differential spots were accumulated in proteome of Pseuodomonas due to the presence of Trichoderma, as new, absent, increased and decreased spots. By comparison of conditions revealed 2 protein spots that detected by MS, have newly expressed in present of Plant and Trichoderma. These proteins corresponded to arginine deiminase of P. putida GB-1 and Chaperonin GroEL protein of P. putida S16 that their expressions associated to stress condition.The results indicated that the presence of Plant and Trichoderma induces major changes in the protein profile of Pseudomonas
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Skin cancer has a broad and burdensome impact on the health and well-being of Iranian and account for substantial health care costs to the nation. The first most common form of skin cancer is basal cell carcinoma [BCC]; followed by the squamous cell carcinoma [SCC] and the incidence of malignant melanoma [MM] is lower but is fatal. Kaposi sarcoma [KS] is a tumor caused by human herpes virus 8. In this study, prevalence and incidence of four skin cancer [BCC, SCC, MM and KS] was investigated by considering to risk factors include age, sex, skin color, sun exposure levels, Lesion location, occupations and timeout to seek treatment. In this study, 95 patients with skin cancers registered in Loghman Hakim hospital during the 7 years from 1998 to 2004 were analyzed. Result depicted that BCC is the most common skin cancer in both sexes and in all types male incidence was significant. Age prevalence of all was about 50 to 80 years. The most common sites of tumor involvement in BCC and SCC were head and neck; KS was lower limb and MM had sporadic lesions. Almost all of patients referred to diagnosis or treatment 1 to 5 years after the initial onset of the disease. Occupations of the majority of patients with skin cancer were farmers. More patients lived in the area with warm and dry climate. In sum up, the skin cancer risk factors are included older ages, residence of warm and dry regions, be male and farmer, and most importantly rate of exposure to sunlight can influence lifestyle of patients that everyone can easily take to protect in different ways
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Because of the huge amounts of proteomic data and demand for new methods of laboratory analysis results, proteins collective analysis, in addition to taking less time, biostatistician assist at identification of new patterns in the data set. In this study, rat hippocampus proteome in normal and Alzheimer's disease [AD] were analyzed by using proteomic techniques and bioinformatics' analysis. Protein extracts from normal and Alzheimer's rats were separated by using two-dimensional electrophoresis [2DE]. The silver staining method was used for detecting spots. Bioinformatics analysis of proteome were performed by progensis same spots software. Bioinformatics and statistical analysis of 2DE gel techniques obtained 760 protein spots were detected in both normal and AD rats. Comparisons between controls and Alzheimer gel containing 20 common proteins were expressed significantly differences. 16 new proteins were expressed in AD, while 36 proteins were suppressed. Proteins clustering by using correlation analysis evaluated 3 clusters in the proteome; Principal component analysis also confirmed the results of clustering. Finally, we can conclude that a significant expression of Alzheimer changes in the hippocampus proteome which are associated with specific biological processes summarized in 3 main clusters indicated 3 principal biological pathways of AD
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The edible leafy vegetables contain nutritional ingredients that are necessary for human health and it is important that nutrients protection be monitored during processing and storage. The aim of this study was to study some organoleptic attributes of a very popular Iranian meal named Coco-Sabzi, which was prepared with a mixture of edible grinded leafy vegetable pre-stored at different frozen conditions. So, by sensorial evaluation we can conclude about nutritional loss of products. The mixture of five edible grinded leafy vegetables including Allium ampeloprasum var. porrum, Lepidium sativum, M. spicata [M. sativa], Ocimum basilicum and Allium porrum were stored at -9, -12 and -18 C for 120, 150 and 180 days. The organoleptic attribute of the prepared Coco-Sabzi was compared with the above three different time-temperature combinations during the frozen storage period. Results indicated that the best colors were observed at -18, -12 and -9 C, respectively. Taste and overall acceptability at -18oC after 120 and 150 days and also at -12 C after 180 days ranked 1st [P< 0.05]. Data analysis showed that the color, taste and overall acceptability of samples were not statistically different at three different time-temperature combinations during the frozen storage period. As a result, organoleptic attribute during six months of frozen storage was affected by freezing temperature, not by frozen storage period
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There are several types of cancer, which cause millions of deaths worldwide every year. Many studies have confirmed that plants are adequate natural sources to be examined as anti-cancer drugs with fewer side effects than chemotherapy and radiotherapy. In this study the anti-cancer properties of Lavender aqueous extract on lymphocytes derived from patients with Hodgkin's lymphoma has been studied. In order to determine the cytotoxic effects of the extract on lymphocytes of patients in stages III and IV of Hodgkin's lymphoma and two different cell lines in the presence of different concentrations of aqueous extract of Lavender, MTT colorimetric assay and flow cytometry analysis were used. Findings indicated that Lavender inhibited cell proliferation in both lymphocytes and cell lines with different effects. The effective concentration of Lavender that decreased viability of Hodgkin's lymphoma cells below Lethal Concentration 50 [LC50] value was 100 micro g/ml and this was half of the therapeutic dose. In addition, apoptosis was the main mechanism the Hodgkin's lymphoma cell encountered when exposed to the aqueous extract of Lavender. Conclusion: This experiment proposes that aqueous Lavender extract can be regarded as a potential anti-cancer agent in future studies
Subject(s)
Humans , Plant Extracts , Hodgkin Disease , Lymphocytes , Cell Proliferation , Cell Line , CytotoxinsABSTRACT
In this paper effect of combinational usage of calprotectin and etoposide on AGS cell line is studied. Application of combined toxic agents such as etoposide and cicplatin are commonly used for chemotherapy purposes. As a matter of fact, calprotectin and etoposide were both applied on human gastric adenocarcinoma cell line [AGS] as antitumor agents. Both calprotectin and etoposide are topo II inhibitor. Etoposide is a lipophilic agent that can easily transport from membrane while calprotectin active intracellular pathway, probably by membrane surface receptor. Calprotectin was purified from human neutrophil by chromatography methods. The human gastric adenocarcinoma cell line was exposed to different concentrations and combinations of calprotectin and etoposide. MTT assay was applied for evaluation of cytotoxicity assay. Viability of AGS cell line was reduced in high dosages of calprotectin and etposide. In fact, overnight incubation of these two agents together has been shown less effective than individual usage. The result indicates that, the combination of both calprotectin and etoposide is considerably less cytotoxic on gastric cancer cells [AGS] than applying individually
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In this study the anticancer activity of Lavender aqueous extract against MKN45 cell line was evaluated. Plant-based drugs are regarded as promising therapies. Lavender is a plant that has been cultivated from ancient times. An aqueous extract of Lavender has shown therapeutic effects on the nervous system in the high doses based on in-vivo studies. Gastric cancer is one of the frequent cancers in Iranian population. We therefore assessed the effect of Lavender upon a gastric cancer cell line. The MKN45 cancer cell line was selected for treatment with aqueous extract of Lavender Survival of MKN45 cell line was studied in the presence of various concentrations of Lavender extract by MTT assay method. Morphological studies were performed via microscopic analyses. Flow cytometry and proteomics techniques were applied to determining pharmaceutical mechanism of lavender cytotoxic effects. The survival and morphological studies revealed anticancer characteristics of extract. Flow cytometry findings indicate that Lavender extract had a cytotoxic effect upon the cell line. Proteomics analysis identified a significant spots showed changes in protein expression levels by informatics analysis. Of the proteins, expression of three cancer biomarkers, Annexin1, Anolase1 and HSP70 were suppressed by extract. This study suggest that Lavender extract is cytotoxic and alter protein expression in a gastric cancer cell line
Subject(s)
Stomach Neoplasms/therapy , Cell Line, Tumor/drug effects , Plant Extracts/pharmacology , Biomarkers , Plants, MedicinalABSTRACT
Esophagus cancer is the eighth most common cancer worldwide and particularly high in an area extending from the southern border of the Caspian Sea in Iran across central Asia to China. Since information about this mysterious disease is poor, proteomics may be solving this enigma. Altering gene expression in cancer cell is a remarkable indicator can be detected by proteomics techniques and bioinformatic analysis. In this study, normal and cancerous cells were obtain from patients, total proteins were purified by standard methods, and proteins separated by two dimensional electrophoresis [2DE]. Some of proteins were identified by Mass spectrometry [MS-MALDI method]. By using bioinformatic analysis illustrate molecular mechanism in this disease. Analysis of gels base on Flicker software and Mass Spectrometry led the same result. 61 protein spots detected in both gels that 21 spots have down regulated and 12 spots have up regulated in cancerous cell than normal. About 14 spots were disappeared in cancer cell while 14 new spots expressed. By using flicker detected 8 Protein that refer to TRFE, SZ07, C1 TC, Kininogen, anexin, keratin, fructosebisphosphate aldolase A and heat shock. Mass spectrometry [MS-MALDI method] identified anexin, keratin, fructosebisphosphate aldolase A and heat shock. Identified proteins were functionally categorized based on Gene Ontology [GO] annotation terms using the DAVID program package. The major molecular functions that annotated with PIR include phosphoprotein, disease mutation while annotated by GO include response to organic substance, response to wounding and cellular homeostasis. The cellular component and molecular function presenting the greatest enrichment that concluded two clusters that the two most importants are cellular homeostasis and extracellular region part. Results reveal that the most of molecular function in cancerous tissue maintenance cellular homeostasis, cell regeneration and repair, so tissues undergo stress try to survive. It can be also concluded that aldolase A, fructose-bisphosphate, keratin 14, formyltetrahydrofolate synthetase and transferrin can be some diagnostic biomarkers and also drug targets in esophagus cancer
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One of the prominent types of connective tissue cells is fibroblast that synthesizes and maintains the extracellular matrix of many animal tissues. Previous studies illustrated that calprotectin protein has different cytotoxicity effects on fibroblast cells. Calprotectin is abundant in the mneutrophil cytosol; it has growth-inhibitory and apoptosis-inducing activities against various mcell types such mas tumor cells. The present study tries to introduce mechanism of growth inhibitory effect of calprotectin on human foreskin fibroblast cells [HFFF] and compare to etoposide [chemotherapy agent as control]. Calprotectin was purified from human neutrophil by chromatography methods. HFFF cell lines were used, maintained in RPMI 1640 medium supplemented with 10% FCS in a humidified incubator [37 degreeC and 5% CO2]. The HFFF cells were exposed to the different concentrations of calprotectin and etoposide for 24, 48 and 72 hours. Cell proliferation was assessed by using dimethylthiazol diphenyl tetrazolium bromide assay. Flow cytometric analysis was performed to evaluate the cytotoxic mechanism of calprotectin on HFFF cells. Our results revealed that calprotectin and etoposide induce growth inhibition of HFFF in dose- and time-dependent manners. Sensitivity of HFFF cells to cytotoxic effect of human calprotectin was highly remarkable. In addition, growth inhibitory effect of this cytotoxic agent mostly was governed through induction of apoptosis in the HFFF cells. Taken together, calprotectin not only has more potent anticancer activity in comparison with the etoposide, but it also is an apoptosis inducer that acts on the proliferation of normal cells like fibroblasts
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Aldolase C as a glycolytic enzyme is associated with cellular structure at developmental stages of all cells, and this is particularly evident during the early stages of morphogenesis. It seems that expression of aldolase C can be regulated by the rate of differentiation that depends on the level of transcription or mRNA stability. There are several techniques to detect gene expression here proteomics was used for determining expression of aldolase C [as a differentiation factor] in several cell types and basal cell carcinoma [BCC] tissue. The human astrocytes were differentiated from mesenchymal stem cells, fibroblast cells were cultured as primary cell culture and BCC tissue was taken from the patient. The fibroblast cells divided into two groups including sham and exposed groups. The exposed cells are them that were exposed to continue Extremely Low-Frequency Electromagnetic Fields [ELF-EMF]. The analysis of 2DE gels, showed different expression of aldolase C in mentioned cells. The findings indicate that the amount of aldolase C expression decreases as differentiation process develops
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Human fibroblasts are the part of the dermis that secrete extracellular matrix for the purpose of tissue repair. Culturing fibroblasts, which leads to formation of a monolayer of these cells, is used for treating various conditions including thermal burns and other skin defects such as diabetic and varicose vein leg ulcers. Therefore, we aimed at developing a fibroblast bank to accomplish multiple goals including skin repair in defects such as burns and ulcers and also performing various research projects on these cells in order to further study of the mechanisms involved in wound healing, rejuvenation and medication effects. We initially developed primary cultures of skin fibroblasts in a DMEM medium. These primary cultures were formed by washing and trypsinizing foreskin specimens followed by separation of epidermis from dermis and cutting the dermis into small pieces. In about 10 days, a monolayer of fibroblasts was formed. We were able to develop the fibroblast bank successfully and to initiate other projects utilizing this bank. With these cultured cells, we would be able to perform different research projects including studying the mechanisms of wound healing, rejuvenation, drug affects, inflammatory mediators, growth factors, etc. Moreover, further progress in this field will result in our independence from requesting these cells from external sources