A method for quantitation of collagen was established by detecting marker peptide with high performance liquid chromatography-mass spectrometry (HPLC-MS). Theoretical marker peptides were selected by sequence comparison. Bovine collagen type I was digested with trypsin. Marker peptides typical for collagen type I were identified with HPLC-MS. The relationship between the abundance of marker peptides and collagen concentration was established. The results show that GEAGPSGPAGPTGAR and the other 5 peptides showed high resolution during chromatographic separation and high signal intensity during MS analysis. Peptide signal intensity and collagen concentration showed a good linear relationship in the range from 0.1 to 3 mg/mL. Bovine tendon and collagen sponge were used as actual samples and collagen contents were determined as 90.2% and 93.4% respectively. Quantitation of marker peptides of collagen was a feasible method to identify and quantify collagens in medical device research and development.
Animals ; Cattle ; Chromatography, High Pressure Liquid ; Collagen Type I ; analysis ; Mass Spectrometry ; Peptides ; analysis

OBJECTIVETo optimize the preparation technology of effective fraction of Catharsius molossus, and investigate the feasibility of process control by the physical and chemical characterization of extracts.

METHODUsed single-factor test method, choosed the main effective components of peptides and amino acids as indexes, combined with theology, chemistry, electricity, and other characterization, the study researched the prepared technology of effective fraction of C. molossus including extraction, concentration, separation, purification, drying and so on.

RESULTThe optimal preparation technology of effective fraction of C. molossus was that soaked an amount of crude drugs with three times of 85% ethanol for 48 h, added 10 times of 85% ethanol, percolated in 4 mL x min(-1) x kg(-1), collected percolation liquid, concentrated to 1:1 at 50-55 degrees C, removed fat by frozen, adopted DA201-C macroporous resin, used 1 BV of water and 4 BV of 70% ethanol as eluting agent, collected eluant respectively. The water part was concentrated and dried, then washed twice with 85% ethanol, collected washing liquid and mixed with 70% ethanol eluant. The product was obtained by concentrating and dring. At the same time, the liquid-phase system of each link was characterized in preparation of effective fraction of C. molossus, which showed that the surface tension related to polypeptide was essentially unchanged, and the conductivity related to salt decreased by about 90% with ineffective substances closely related to salt The results showed that the preparation technology maximumly retained the effective information, and removed the invalid information.

CONCLUSIONThe preparation technology of effective fraction of C. molossus is stable and reliable, and the process control in physico-chemical characterization of extracts is feasible.

Amino Acids ; analysis ; Animals ; Chemical Fractionation ; Chemical Phenomena ; Chemistry, Pharmaceutical ; Coleoptera ; chemistry ; Peptides ; analysis

An overview of the background for proteomics and a description of the present state of art are given with a description of the main strategies in proteomics. The advantages and limitations of the two major strategies, 2D-gel based and LC-MS based, are discussed and a combination for the two, CeLC-MS is described. A number of challenging problems which have been solved using different proteomics strategies including the advantage of organell enrichment or modifications specific peptide isolation to get deeper into the proteome are described. Finally the present status and future needs discussed.
Chromatography, High Pressure Liquid ; Electrophoresis, Gel, Two-Dimensional ; Mass Spectrometry ; Peptides ; analysis ; chemistry ; Phosphorylation ; Protein Isoforms ; analysis ; Proteome ; analysis ; Proteomics

Pterygium is a proliferative disease. Recent research has reported that stem cells are involved in the pathogenesis of various proliferative diseases, including solid tumors and diabetic proliferate vitreoretinopathy. In previous literature, we hypothesized that adult stem cells originated from bone marrow were involved in the pathogenesis of pterygium. We proved this by immunohistochemical staining with various stem cell markers. The staining showed adult stem cells in the pterygium. c-kit positive cells were observed primarily in the stroma, and some cells were also found in the basal epithelium. AC133 and CD34 positive cells were primarily found in the basal epithelium and were ovoid shaped, similar to the c-kit cells. However, some cells were found in vascular endothelium. STRO-1 positive cells were found mainly in the stroma and were spindle shaped. In recurrent pterygium, cells were more scattered and the expression pattern was denser. Therefore, we suggest a new theory of pterygium pathogenesis. Inflammation caused by environmental factors triggers the abnormal production of some growth factors and cytokines in order to recover from cellular damage. If these healing signals are excessive, limbal basal cells will be changed to abnormally-altered pterygial cells. The excessive wound healing process and remnant altered cells result in recurrence using the same mechanism.
Stem Cells/*physiology ; Pterygium/*etiology ; Proto-Oncogene Proteins c-kit/analysis ; Peptides/analysis ; Middle Aged ; Humans ; Glycoproteins/analysis ; Bone Marrow Cells/*physiology ; Antigens, CD34/analysis ; Antigens, CD/analysis

BACKGROUND/AIMS: Preoperative N-terminal pro-brain natriuretic peptide (NT-proBNP) is a useful predictor of postoperative cardiovascular complications. The present study investigated whether blood NT-proBNP values are suitable for predicting postoperative cardiovascular complications after non-cardiac surgery in elderly patients showing normal left ventricular (LV) function on preoperative echocardiograms. METHODS: This study was performed by analyzing the medical records of elderly patients referred to the cardiology department for the purpose of assessing their cardiac function before orthopedic surgery. Of the patients who underwent echocardiography and NT-proBNP assessment simultaneously, 275 patients aged > or = 70 years and with an LV ejection fraction of > or = 55% were included in the study. RESULTS: Major adverse cardiac and cerebrovascular events (MACCEs) occurred in 33 (12%) of the 275 patients, and the NT-proBNP concentration was higher in patients with complications than in those without complications (1,904.20 +/- 2,300.23 vs. 530.58 +/- 882.27 pg/mL, p < 0.01). The ROC area under the curve was 0.756 (95% confidence interval 0.701-0.805, p < 0.001) with an optimal cutoff of 416.3 pg/mL (69.7% sensitivity, 67.36% specificity). A multivariate analysis showed that a preoperative age of > 80 years (odds ratio, 2.313; p = 0.047) and an increased blood NT-proBNP concentration (odds ratio, 3.189; p = 0.009) were independent risk factors for the prediction of MACCEs. CONCLUSIONS: Although elderly patients scheduled to undergo non-cardiac surgery may show normal LV systolic function on echocardiography, measurement of their preoperative blood NT-proBNP concentration is useful for predicting MACCEs occurring after non-cardiac surgery.
Aged* ; Cardiology ; Echocardiography ; Humans ; Medical Records ; Multivariate Analysis ; Natriuretic Peptides ; Orthopedics* ; Risk Factors

OBJECTIVETo identify and characterize the polypeptides specifically binding to human B type natriuretic peptide (BNP) screened from 12TM phage display peptide library.

METHODSThe BNP-binding peptides were screened from 12TM phage display peptide library and identified by ELISA.

RESULTSAfter 4 rounds of screening, 10 of the 16 phage clones were identified as the positive clones which could bind to BNP. Five amino acid sequences were obtained in the 10 positive clones. Dose-dependent ELISA results demonstrated that the screened polypeptides could specifically bind to BNP.

CONCLUSIONThese screened polypeptides can bind specifically to BNP, which provides a basis for further research on expression and purification of anti-BNP polypeptides and the development of the detection kit of BNP.

Humans ; Natriuretic Peptide, Brain ; metabolism ; Peptide Library ; Peptides ; analysis ; isolation & purification ; metabolism ; Protein Binding

OBJECTIVETo separate and identify cyclopeptides of tubers of Rubia schumanniana.

METHODThe 70% methanol extracts from tubers of Rubia schumanniana were separated and purified by silica gel, RP-18, Sephedax LH-20 and HPLC. Their structures were identified by spectral analysis.

RESULTNine cyclopeptides were separated and identified as RA- II (1), RA-V (2), RA-VIII (3), rubiyunnanin C (4), RA-X (5), RY-II (6), RA- I (7), RA-XIII (8) and RA-XIII-OMe (9), respectively.

CONCLUSIONAll of nine cyclopeptides were separated from R. schumanniana for the first time.

Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Peptides, Cyclic ; analysis ; isolation & purification ; Rubia ; chemistry

Small proteins (SPs) are defined as peptides of 100 amino acids or less encoded by short open reading frames (sORFs). SPs participate in a wide range of functions in cells, including gene regulating, cell signaling and metabolism. However, most annotated SPs in all living organisms are currently lacking expression evidence at the protein level and regarded as missing proteins (MPs). High efficient SPs identification is the prerequisite for their functional study and contribution to MPs searching. In this study, we identified 72 SPs and successfully validated 9 MPs from Saccharomyces cerevisiae based on SPs enrichment strategy. In-depth analysis showed that the missing factors of MPs were low molecular weight, low abundant, hydrophobicity, lower codon usage bias and unstable. The small protein-based enrichment can be used as MPs searching strategy, which might provide the foundation for their further function research.
Codon ; Open Reading Frames ; Peptides ; Saccharomyces cerevisiae ; Saccharomyces cerevisiae Proteins ; analysis

The acquisition of somatic mutations is the most common event in cancer. Neoantigens expressed from genes with mutations acquired during carcinogenesis can be tumor-specific. Since the immune system recognizes tumor-specific peptides, they are potential targets for personalized neoantigen-based immunotherapy. However, the discovery of druggable neoantigens remains challenging, suggesting that a deeper understanding of the mechanism of neoantigen generation and better strategies to identify them will be required to realize the promise of neoantigen-based immunotherapy. Alternative splicing and RNA editing events are emerging mechanisms leading to neoantigen production. In this review, we outline recent work involving the large-scale screening of neoantigens produced by alternative splicing and RNA editing. We also describe strategies to predict and validate neoantigens from RNA sequencing data.
Alternative Splicing ; Carcinogenesis ; Humans ; Immune System ; Immunotherapy ; Mass Screening ; Peptides ; RNA Editing ; RNA ; Sequence Analysis, RNA

A Bacillus sp. BS061 significantly reduced disease incidence of gray mold and powdery mildew. To identify the active principle, the culture filtrate was partitioned between butanol and water. The antifungal activity against B. cinerea was evident in the butanol-soluble portion, and active substances were identified as cyclic lipopeptides, iturin A series, by nuclear magnetic resonance spectrometry (NMR) and mass analysis. Interestingly, antifungal activity against powdery mildew was observed in the water-soluble portion, suggesting that cyclic lipopeptides have no responsibility to suppress powdery mildew. This finding reveals that biocontrol agents of Bacillus origin suppress gray mold and powdery mildew through the secretion of different bioactive substances.
Bacillus* ; Fungi* ; Incidence ; Lipopeptides ; Magnetic Resonance Spectroscopy ; Peptides, Cyclic ; Spectrum Analysis ; Water