The Cytotoxic Effect of Small and Large Molecules of PMF Fraction Extracted from Camel Urine on Cancer Cells.

Aim of the work: Animal urine, including that of camels, has long been used for the therapeutic management of human ailments. In this study, we sought to characterize the cytotoxic properties of newly derived purified fractions from previously described camel urine extract (PMF) on various cancer cell lines. Methodology: Two new size dissimilar fractions of PMF (large and small) were obtained by fractionalizing PMF using 3kD and 50kD membrane filters. A SRB cytotoxicity assay of the PMF fractions was performed on cancer cell lines (A549, HCT116, HepG2, MCF-7, U251 and Hela) as well as normal cell lines (human fibroblast cell line and Vero). Results: This study showed that the newly derived and more purified fraction of PMF (new PMF) possesses effective and selective anti-cancer properties against several types of cancer cell lines. Conclusion: This study, as well as previous ones, suggests that camel urine extracts (old and new PMF) may provide newer therapeutic alternatives to clinically manage cancer patients. However, further studies are needed to verify these positive preliminary results.

Influence of pulsed magnetic field on soybean (Glycine max L.) seed germination, seedling growth and soil microbial population.

The effects of pulsed magnetic field (PMF) treatment of soybean (Glycine max L. cv CO3) seeds were investigated on rate of seed germination, seedling growth, physico-chemical properties of seed leachates and soil microbial population under laboratory conditions. Seeds were exposed to PMF of 1500 nT at 0.1, 1.0 10.0 and 100.0 Hz for 5 h per day for 20 days, induced by enclosure coil systems. Non-treated seeds were considered as controls. All PMF treatments significantly increased the rate of seed germination, while 10 and 100 Hz PMFs showed the most effective response. The 1.0 and 10 Hz PMFs remarkably improved the fresh weight of shoots and roots, leaf area and plant height from seedlings from magnetically-exposed seeds compared to the control, while 10 Hz PMF increased the total soluble sugar, total protein and phenol contents. The leaf chlorophyll a, b and total chlorophyll were higher in PMF (10 and 100 Hz) pretreated plants, as compared to other treatments. In addition, activities of α-amylase, acid phosphatase, alkaline phosphatase, nitrate reductase, peroxidase and polyphenoloxidase were increased, while β-amylase and protease activities were declined in PMF (10 Hz)-exposed soybean plants. Similarly, the capacity of absorbance of water by seeds and electrical conductivity of seed leachates were significantly enhanced by 10 Hz PMF exposure, whereas PMF (10 Hz) pretreated plants did not affect the microbial population in rhizosphere soil. The results suggested the potential of 10 Hz PMF treatment to enhance the germination and seedling growth of soybean.

A primary study of the different protein expression profiles between human oral squamous cell carcinoma and normal oral mucosa tissues / 实用口腔医学杂志

Objective: To identify the different protein expression profiles between human oral squamous cell carcinoma (OSCC) and normal oral mucosa tissues, and provide experimental data for further study of the development mechanism of OSCC. Methods: 10 cases of OSCC and paired normal oral mucosa tissues were collected and analyzed through two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Results: (1) The average protein spots of OSCC were 2 325±390, while that of normal oral mucosa tissues were 2 487±281. (2) 29 differential protein spots were found between OSCC and normal oral mucosa. Moreover, these protein spots were all down- regulated in OSCC compared with normal oral mucosa. Among these spots, 3 were identified as fibrin beta, triosephosphate isomerase (TIM) and unknown protein through mass spectrometry and bioinformation. Conclusion: Down-regulation of fibrin beta, Triosephosphate isomerase(TIM) and unknown protein are found in the development of OSCC and the mechanism needs further study.

Protein Profile Changes in Platelet Concentrates According to Storage and Leukoreduction- Analysis Using Proteomics Technology / 대한진단검사의학회지

BACKGROUND: Knowing how the protein profile of platelet products changes with storage or leukoreduction may give us greater insight into cell physiology and the cause of transfusion reactions other than cytokines and chemokines. METHODS: We filtered four packs of platelet concentrates (PC) within 24 hr of blood collection and after 120 hrs of storage. Four aliquots of each supernatant in PC were obtained: pre-storage+prefiltration, pre-storage+post-filtration, post-storage+pre-filtration and post-storage+post-filtration. Routine chemistry tests and a two-dimensional electrophoresis (2-DE) were performed. The stained images were analyzed and the significant spots were identified using a peptide mass finger printing (PMF) with matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis after trypsin digestion. RESULTS: The protein spots increased with storage and decreased after filtration (P<0.05, prestorage+post-filtration). The spot density of various proteins, including macrophage inflammatory protein-2 alpha, megakaryocyte colony stimulating factor and interleukin-22 changed with storage and leukoreduction. CONCLUSIONS: The database of identified protein spots and their changes produced in this study is a useful basic tool for future studies on the mechanism of transfusion reactions. Further studies should validate the significance of each protein spot.

Optimization of the major parameters of MS-Fit in peptide mass fingerprint identify proteins / 重庆医科大学学报

Objective:To optimize the major parameters of bioinformatics analysis conditions in process of PMF(Peptide Mass Fingerprinting) identify target proteins. Methods:In this process of experiment design and analysis,we made the Bovine Carbonic anhydrase-2 and BSA from 2-DE gel,after proteolysis with porcine trypsin and gathered the peptides,subsequently identified with MALDI-TOF MS and got the peptides mass data. Then,we selected database of Swissprot and MS-Fit search engine,the standard Carbonic anhydrase-2 protein used as model to set up protein search standard conditions. Results:Our study showed that the cysteine was modified by carbamidomethylation more convenient in protein search,at the same time,the better mode of mass tolerance was percentage and 0.1% was suitable tolerance in our search. Then we used BSA mass data to check the accuracy of MS search standard conditions. Conclusions:Results show that the optimized parameters are reliable.

2D-PAGE Analysis of Chinese Rose Leaf Protein Under Heat Shock Stress / 中国生物工程杂志

Proteins extracted from two varieties of Chinese roses leaves were separated by two- dimensional polyacrylamide gel electrophoresis (2-DE) with immobilized pH gradient (IPG). Many difference proteins were isolated with molecular weights ranging 10-30 kDa and pI5-6. Three proteins of high levels observed in a gel were excised and identified using peptide mass fingerprinting and MS-MS. A summary of the identified proteins and their putative functions are presented. They are identified as eIF-5A、LEA protein and Hsp17. 5. Functions of these proteins in plant tolerance to high temperature were discussed.

Screening of differential expression proteins from human oral lichen planus and normal oral mucosa by two-dimensional difference gel electrophoresis and mass spectrometry / 实用口腔医学杂志

Objective:To find biomarkers for oral lichen planus by comparing differential expressing proteins. Methods:10 cases of oral lichen planus and normal oral mucosa tissues were collected.Total protein was extracted; differential proteome profiles were established and analyzed by two-dimensional polyacrylamide gel electrophoresis(2D-PAGE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Results:(1)The well-resolved,reproducible 2-DE patterns of oral lichen planus and normal oral mucosa were obtained. The results showed that average protein spots were 1 576?67 and 1 608?73 in oral lichen planus and normal oral mucosa respectively, (2) The 13 differential protein spots were identified by Imaging Master 2D image analysis software between oral lichen planus and normal oral mucosa. There were 7 protein spots in oral lichen planus were higher than those in normal oral mucosa, 6 protein spots in oral lichen planus were lower than those in normal oral mucosa. 10 differential expressing proteins were analyzed by mass spectrometry and bioinformation. 4 of them were well characterized including manganese superoxide dismutase (Mn-SOD), Annexin I, vimentin and unknown proteins. Conclusion:Differential expression proteins might be candidate biomarkers for diagnosis of oral lichen planus;and proteomic technique is valuable for screening the diagnostic biomarkers.

MALDI-TOF MS identification of protein of mitochondria from cultured hepatoma cells / 重庆医科大学学报

Objective:To explore the optimal conditions associated with proteolytic digestion in 2-DE gel,PMF analysis and database inquiry,and to construct a mass identification strategy in research for mitochondrial comparative proteomics.Methods:The BSA standard protein was incised from 2-DE gel by Coomassie Brilliant Blue dyeing and was mixed with matrix (CHCA),then analyzed by MALDI-TOF MS.The differentially expressed mitochondria protein L9,an unknown protein obtained from hepatoma cells QGY-7703,was identified by condition with BSA optimizing.The PMF was inquired by MS-Fit database.Results:The BSA was identified in Masses Matched number 10/11 and Sequence Overcast 19% of PMF,which proved to the reliability of experimental conditions.By database Inquiry about PMF from L9 protein,OXCT(3-oxoacid CoA transferase 1 precursor)was identified with Masses Matched number 9/13 and Sequence Overcast 24%.The OXCT was in accord with position on gel.Conclusion:This method is applicable to research for mitochondrial comparative proteomics.