Identification of Antimicrobial and Protective Protein Distribution in Salivary Protein Complexes via Gel Filtration High Performance Liquid Chromatography

2016 ◽  
Vol 40 (6) ◽  
pp. 891-898
Author(s):  
Yeon Sook Kim ◽  
◽  
Suk Keun Lee
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Protein Complexes ◽  
Gel Filtration ◽  
Salivary Protein ◽  
Protein Distribution

Aspirin Preferentially Inhibits Arachidonic Acid Metabolism In Collagen Vs. Thrombin-Stimulated Platelets

1981 ◽  
Author(s):  
D Deykin ◽  
R Vaillancourt
Keyword(s):  
High Performance Liquid Chromatography ◽  
Arachidonic Acid ◽  
Liquid Chromatography ◽  
High Performance ◽  
Acid Metabolism ◽  
Gel Filtration ◽  
Human Platelets ◽  
Arachidonic Acid Metabolism ◽  
Selective Effect ◽  
Oxygenase Activity

The purpose of this study was to compare the effect of aspirin on the release of metabolites of arachidonic acid from thrombin and collagen stimulated platelets. Human platelets were incubated with tritium-labeled arachidonic acid and then isolated by gel filtration. The labeled platelets were stimulated with varied doses of either thrombin or collagen for 15 minutes. The platelets were then pelleted and the released metabolites of arachidonic acid were separated by high-performance liquid chromatography. In experiments with aspirin, the aspirin was added 5 minutes before either thrombin or collagen. The total release of radioactivity was comparable at 15 μg/ml of collagen and 1.0 units/ml of thrombin (approximately 10% of the total) and at 100 μg/ml of collagen and 5 units/ml of thrombin (approximately 30%). Aspirin (25 μg/ml) preferentially inhibited collagen-stimulated release of radioactivity (62% inhibition of release with 15 μg/ml of collagen vs. 25% inhibition of release with 1.0 units/ml of thrombin; 54% inhibition of release with 100 μg/ml of collagen vs. 8% inhibition of release with 5.0 units/ml of thrombin). At all concentrations of collagen or thrombin, cyclo-oxygenase activity was markedly reduced by aspirin. The selective effect of aspirin on collagen reflects primarily preferential suppression of HETE formation. We conclude that aspirin inhibits the formation of both lipoxygenase and cyclooxygenase-derived products in collagen-stimulated platelets.

The use of reversed-phase high-performance liquid chromatography to detect proteolytic activity from human pancreas in a radioimmunoassay for corticotrophin-releasing factor

1987 ◽  
Vol 114 (1) ◽  
pp. 147-151 ◽  
Author(s):  
D. S. Jessop ◽  
R. L. Patience ◽  
D. Cunnah ◽  
L. H. Rees
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Proteolytic Activity ◽  
High Performance ◽  
Gel Filtration ◽  
Reversed Phase ◽  
Pancreatic Tissue ◽  
Acid Extraction ◽  
Human Pancreas ◽  
Corticotrophin Releasing Factor

ABSTRACT Degradation of tracer during a radioimmunoassay (RIA) can result in false-positive concentrations of immunoreactivity being reported in a biological sample. A technique has been developed using reversed-phase high-performance liquid chromatography (HPLC) to detect proteolytic degradation of corticotrophin-releasing factor-41 (CRF-41) during incubation with tissue extracts under RIA conditions. Human pancreatic tissue was extracted in HCl or urea and incubated with 125I-labelled CRF-41 at neutral pH for 18 h. When samples were analysed by HPLC and fractions counted for radioactivity, tracer was extensively degraded. Heating extracts at 85 °C or adding lima bean trypsin inhibitor to the medium prevented degradation. Pancreatic tissue extracted in HCl was analysed by gel filtration and HPLC, and fractions were subjected to RIA for CRF-41. A peak of immunoreactivity was detected by both chromatographic methods. However, when this material was incubated with tracer and analysed by HPLC, the tracer was degraded, indicating that proteolytic activity remained after acid extraction and two forms of chromatography. J. Endocr. (1987) 114, 147–151

scholarly journals Detection of cheese whey and caseinomacropeptide in fermented milk beverages using high performance liquid chromatography

2014 ◽  
Vol 66 (3) ◽  
pp. 959-964 ◽  
Author(s):  
E.H.P. Andrade ◽  
M.R. Souza ◽  
L.M. Fonseca ◽  
C.F.A.M. Penna ◽  
M.M.O.P. Cerqueira ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Storage Time ◽  
Cheese Whey ◽  
Starter Culture ◽  
Gel Filtration ◽  
Fermented Milk ◽  
Refrigerated Storage ◽  
Four Levels

Cheese whey level and caseinomacropeptide (CMP) index of fermented milk beverages added with four levels of cheese whey (0, 10, 20, and 40%) and stored at 8-10oC for 0, 7, 14 and 21 days were determined by high performance liquid chromatography-gel filtration (HPLC-GF). Additionally, the interference of the starter culture and the storage time on the detection of cheese whey and CMP were investigated. Refrigerated storage up to 21 days did not affect (P>0.05) cheese whey and CMP amounts in milk (0% of cheese whey) and in fermented milk beverages added with 10 and 20% of cheese whey (P>0.05). However, cheese whey and CMP amounts were higher than expected (P<0.05) in fermented milk beverages added with 40% of cheese whey and stored for 21 days.

Isolation of endogenous antimicrobial peptides from blood cells

Bacteriology ◽  
2020 ◽  
Vol 5 (1) ◽  
pp. 33-36
Author(s):  
I.A. Bazikov ◽  
◽  
A.N. Maltsev ◽  
O.I. Sedykh ◽  
V.A. Baturin ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Antimicrobial Peptides ◽  
Blood Cells ◽  
High Performance ◽  
Pore Diameter ◽  
Gel Filtration ◽  
Antibiotic Resistant ◽  
Filter Surface ◽  
Peptide Fractions

A technique for isolating the endogenous antimicrobial peptides (AMP) has been developed. Leukocyte-erythrocyte-platelet blood mass of donors was used as the material for preparing endogenous AMP. The hydrolysate was obtained with the trypsin solution from this mass, and then the components were fractionated using the filter chromatography column (Simax CSN ISO 3585, Russia). For gel filtration, 1.5 g of Sephadex G-25 was applied to the filter surface followed by sterilizing filtration of the obtained substance through bactericidal filters having a pore diameter of 0.2 μm. The antimicrobial peptide fractions were isolated using high performance liquid chromatography. Subsequently, AMP were encapsulated into the organosilicon niosomes. Key words: antimicrobial peptides, defensins, niosomes, antibiotic-resistant microorganisms

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