scholarly journals Identification of food-derived peptides in human blood after ingestion of corn and wheat gluten hydrolysates

2018 ◽  
Vol 2 ◽  
Author(s):  
Akika Ejima ◽  
Megumi Nakamura ◽  
Yasushi A. Suzuki ◽  
Kenji Sato
Keyword(s):  
Matrix Protein ◽  
Leucine Aminopeptidase ◽  
Wheat Gluten ◽  
Protein Hydrolysates ◽  
The Body ◽  
Size Exclusion ◽  
Extracellular Matrix Protein ◽  
Reverse Phase Hplc ◽  
Before And After ◽  
Exclusion Chromatography

Bioactive peptides in the body after ingestion of plant protein hydrolysates have been speculated but not identified. We aimed to establish an approach to identify small amounts of food-derived peptides in humans after ingestion of non-extracellular matrix protein hydrolysates. Corn and wheat gluten hydrolysates were digested using pancreatin and leucine aminopeptidase; the resultant peptides were identified via size-exclusion chromatography and reverse-phase HPLC-tandem mass spectrometry (MS/MS). Structures of indigestible peptides were confirmed via LC-MS/MS in multi-reaction monitoring mode. All indigestible peptides in the exopeptidase digest were diprolyl and di- and tripyroglutamyl peptides. Blood collected from healthy volunteers (n = 4) before and after ingestion of 9 g of the hydrolysates was assessed for the indigestible peptides via LC-MS/MS. Six peptides (Pro-Ala, Pro-Gly, Pro-Gln, pyroGlu-Pro, pyroGlu-Leu-Pro, and pyroGlu-Gln-Pro) significantly increased in human plasma up to 10–100 nM compared to the baseline. This may hence be a powerful tool for identifying foodderived peptides in blood.

scholarly journals Orphan Designation and Cisplatin/Hyaluronan Complex in an Intracavitary Film for Malignant Mesothelioma

Pharmaceutics ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 362
Author(s):  
Sabrina Banella ◽  
Eride Quarta ◽  
Paolo Colombo ◽  
Fabio Sonvico ◽  
Antonella Pagnoni ◽  
...  
Keyword(s):  
Sodium Hyaluronate ◽  
Chloride Ions ◽  
Complete Resection ◽  
Size Exclusion ◽  
European Medicines Agency ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Pharmaceutical Development ◽  
Film Forming ◽  
Exclusion Chromatography

Pleural mesothelioma is a lung diffuse tumor, whose complete resection is unlikely. Consequently, metastases reappear where the primary tumor was removed. This paper illustrates the orphan medicine designation procedure of an intracavitary cisplatin film and related pharmaceutical development aspects requested by the European Medicines Agency (EMA) in its Scientific Advice. Since cisplatin pharmacokinetics from the implanted film in sheep resulted substantially modified compared to intravenous administration, the formation of a cisplatin/hyaluronan complex had been hypothesized. Here, the interaction between sodium hyaluronate (NaHA) and cisplatin (CisPt) was demonstrated. Size exclusion chromatography qualitatively evidenced the complex in the film-forming mixture, only showing the NaHA peak. Atomic absorption spectroscopy of the corresponding fraction revealed platinum, confirming the interaction. Reverse phase HPLC quantified about 5% free cisplatin in the film-forming mixture, indirectly meaning that 95% was complexed. Finally, a study of CisPt release from the film assessed how CisPt/NaHA complex affected drug availability. In water, a medium without chloride ions, there was no release and the film remained intact for 48 h and longer, whereas the placebo film dissolved in 15 min. In 0.9% NaCl medium, the film became more soluble, dissolving within 3–4 h. However, cisplatin release was still controlled by the existing complex in solution until chloride ions displaced it. While the film modified its dissolution with aging, CisPt release remained unaffected (90% released in 48 h).

In vitro digestibility study of thermal oxidized palm oleins Estudio de la digestibilidad in vitro de oleínas de palma termooxidadas

2000 ◽  
Vol 6 (6) ◽  
pp. 449-456 ◽  
Author(s):  
F.J. Sánchez-Muniz ◽  
R. Arroyo ◽  
J.M. Sánchez-Montero ◽  
C. Cuesta
Keyword(s):  
Pancreatic Lipase ◽  
High Performance ◽  
Palm Olein ◽  
Size Exclusion ◽  
In Vitro Digestibility ◽  
Moderate Degree ◽  
Before And After ◽  
Exclusion Chromatography ◽  
Hydrolysis Of

Information on digestibility and absorption of oils and fats used for frying is under debate. To get knowledge on this, unused palm olein (9.27 ± 0.10% w/w polar content), used frying palm olein with a moderate degree of alteration (14.81 ± 0.90% w/w polar content) and highly altered used frying palm olein (26.36 ± 0.30% w/w polar content) and their respective nonpolar and polar fractions were studied. Samples were analyzed by high-performance size-exclusion chromatography before and after a 20-min in vitro incubation with pancreatic lipase. Formation of monoacylglycerols and free fatty acids reflected no relevant differences between unused and moderately altered oleins, whereas the most altered olein was hydrolyzed to a much lesser degree. The presence of oligomers (dimers and polymers of triacylglycerols) negatively affected the hydrolysis of triacylglycerol monomers in whole oleins. The hydrolysis of these monomers in the isolated nonpolar and polar fractions ranked between 60.2% and 78.5%. Oligomers were efficiently hydrolyzed by pancreatic lipase in whole un used and moderately altered oleins but not in the most altered one. Polymers from isolated polar fractions were poorly hydrolyzed or not hydrolyzed at all. These data suggest that whole oleins contained some compounds that increase susceptibility of oligomers to enzymatic hydrolysis and that such compounds were not present in the polar fraction.

scholarly journals Biochemical and Pharmacological Characterization of TLBbar, a New Serine Protease with Coagulant Activity from Bothrops barnetti Snake Venom

2013 ◽  
Vol 2013 ◽  
pp. 1-11
Author(s):  
Magaly Alejandra Brousett-Minaya ◽  
Paulo Aparecido Baldasso ◽  
Salomón Huancahuire-Vega ◽  
Sérgio Marangoni
Keyword(s):  
Serine Protease ◽  
Snake Venom ◽  
Serine Proteases ◽  
Kinetic Studies ◽  
Size Exclusion ◽  
Reverse Phase Hplc ◽  
Pharmacological Characterization ◽  
Coagulant Activity ◽  
Exclusion Chromatography

A thrombin-like enzyme named TLBbar was isolated from Bothrops barnetti snake venom and its biochemical and pharmacological characteristics were determined. TLBbar was purified using size exclusion chromatography and reverse phase HPLC, showing molecular mass of 28750.7 Da determined by mass spectrometry. TLBbar serine protease is basic (pI 7.4) and its structure shows similarity with other serine proteases of snake venom. Optimal proteolytic activity was at 37°C and pH 8; this activity was strongly inhibited by PMSF and Leupeptin, however; heparin, and soybean trypsin inhibitor (SBT-I) were ineffective. Kinetic studies on BApNA chromogenic substrate have revealed that TLBbar presents a Michaelis-Menten kinetics, with values of Km and Vmax of 0.433 mM and 0.42 nmol/min, respectively. TLBbar showed high clotting activity upon bovine and human plasma, presenting IC of 125 and minimum dose coagulant (MDC) of 2.23 μg/μL. TLBbar cleavages the Aα chain of bovine fibrinogen, with maximal efficiency at 30–40°C in the presence of calcium after two hours incubation; this fibronogenolityc activity was inhibited by PMSF and Leupeptin, confirming its classification in the group of serine proteases. In addition, TLBbar is capable of aggregating platelets in the same way that thrombin in concentrations of 2.5 μg/μL.

scholarly journals Purification of bacteriocins using size-exclusion chromatography

2016 ◽  
Vol 11 (2) ◽  
pp. 281 ◽  
Author(s):  
Vivek K. Bajpai ◽  
Irfan A. Rather ◽  
Alshammari Fanar Hamad
Keyword(s):  
Mass Spectrometry ◽  
Ammonium Sulfate ◽  
Size Exclusion Chromatography ◽  
Size Exclusion ◽  
Ammonium Sulfate Precipitation ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Cell Free Supernatant ◽  
Video Clips ◽  
Exclusion Chromatography

<p>The bacteriocin purification involves following main steps. a). Extraction of cell-free-supernatant of bacteria. b). Ammonium sulfate precipitation. c). Dialysis. d). Diafiltration using PVP and e). Size-exclusion chromatography. However, depending on the nature of work, the compound could be further analyzed by reverse-phase HPLC, NMR, mass spectrometry and sequencing.</p><p><strong>Video Clips</strong></p><p><a href="https://youtube.com/v/u1BmWfOTS9w">Part 1</a>: 4 min 52 sec</p><p><a href="https://youtube.com/v/aF45JFnwErc">Part 2</a>: 1 min 47 sec</p>

scholarly journals Comprehensive Profiling of Plasma Exosomes Using Data-Independent Acquisitions – New Tools for Aging Cohort Studies

2021 ◽  
Author(s):  
Sandip K. Patel ◽  
Roland Bruderer ◽  
Nathan Basisty ◽  
Joanna Bons ◽  
Pierre-Yves Desprez ◽  
...  
Keyword(s):  
Reversed Phase ◽  
The Body ◽  
Size Exclusion ◽  
Biomarkers Of Aging ◽  
Age Related ◽  
Complex Biological Process ◽  
High Throughput Method ◽  
Aging Study ◽  
Exclusion Chromatography ◽  
Using Data

AbstractAging is a complex biological process associated with progressive loss of physiological function and susceptibility to several diseases, such as cancer and neurodegeneration. Exosomes are involved in many cellular signaling pathways, and their cargo may serve as promising disease or aging biomarkers. These membrane-bound extracellular vesicles facilitate the transport of intracellular contents to proximal and distal cells in the body. Here, we investigated two omics approaches for exosome analysis. To overcome the challenges of plasma exosome contamination with abundant soluble plasma proteins, we developed a high-throughput method to isolate highly purified exosomes from human plasma by sequential size-exclusion chromatography and ultrafiltration. First, we used data-dependent acquisitions from offline high-pH reversed-phase fractions of exosome lysate to generate a deep spectral library comprising ∼2,300 exosome proteins. Second, in a pilot aging study, we used comprehensive data-independent acquisitions to compare plasma exosomes from young (20–26 yrs) and old (60–66 yrs) individuals. We quantified 1,318 exosome proteins, and levels of 144 proteins were significantly different in young and old plasma groups (Q<0.05 and >1.5-fold change). We also analyzed exosome miRNA cargo and detected 331 miRNAs. Levels of several were significantly different in young and old individuals. In addition, 88 and 17 miRNAs were unique to old and young individuals, respectively. Plasma exosome biomarkers have great potential for translational studies investigating biomarkers of aging and age-related diseases and to monitor therapeutic aging interventions.

Purification Method of Silver Nanoparticles (AgNPs) and its Identification Using UV-Vis Spectrophotometer

2020 ◽  
Vol 840 ◽  
pp. 484-491
Author(s):  
Umi Nur Sholikhah ◽  
Deni Pranowo ◽  
Rizky Ibnufaatih Arvianto ◽  
Endang Sarmini ◽  
Triani Widyaningrum
Keyword(s):  
Silver Nanoparticles ◽  
Size Exclusion Chromatography ◽  
Chemical Reduction ◽  
Size Exclusion ◽  
Chemical Reduction Method ◽  
Purification Method ◽  
Resonance Characteristic ◽  
Before And After ◽  
Exclusion Chromatography ◽  
Centrifugation Method

The development of nanotechnology applications is rapidly growing in many sectors. One of them is silver nanoparticles (AgNPs) which are metal nanoparticles that play an important role, especially in nanomedicine. The most effective method of purifying to obtain stable AgNPs is very important to study. Experiments on the separation of AgNPs have been carried out using the size exclusion chromatography and centrifugation methods to see the effectiveness of refining the two methods. This experiment begins with the synthesis of AgNPs using the chemical reduction method. Then, the synthesized AgNPs were purified by Size Exclusion Chromatography (SEC) and centrifugation method then analyzed using UV-Vis spectrophotometer to determine the maximum peaks before and after purification. The experimental results were obtained that centrifugation methods and SEC having the same effectiveness in refining AgNPs. The centrifugation method at various speed (0, 3000, 6000, 9000, 12000 and 15000 rpm) gave wavelength results 403, 404, 404, 405, 404, and 404 nm. The SEC method using Sephadex-25 column showed the 4th to 8th fractions gave the maximum wavelength 404, 404, 404, 405, and 404 nm, respectively. The maximum wavelength of both methods showed the surface plasmon resonance characteristic of AgNPs. However, centrifugation at 3000 rpm has better homogeneity than SEC method.

scholarly journals Influenza A M2 Channel Oligomerization is Sensitive to its Chemical Environment

2021 ◽  
Author(s):  
Julia A Townsend ◽  
Henry M Sanders ◽  
Amber D Rolland ◽  
James S Prell ◽  
Jun Wang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Drug Targets ◽  
Influenza A ◽  
Matrix Protein ◽  
Antiviral Drug ◽  
Native Mass Spectrometry ◽  
Chemical Environment ◽  
Size Exclusion ◽  
Oligomeric State ◽  
Exclusion Chromatography

Viroporins are small viral ion channels that play important roles in the viral infection cycle and are proven antiviral drug targets. Matrix protein 2 from influenza A (AM2) is the best characterized viroporin, and the current paradigm is that AM2 forms monodisperse tetramers. Here, we used native mass spectrometry, ion mobility spectrometry, and size-exclusion chromatography to characterize the oligomeric state of full-length AM2 in a variety of different pH and detergent conditions. Unexpectedly, we discovered that AM2 formed a range of different oligomeric complexes that were strongly influenced by its local chemical environment. The monodisperse tetramer was only observed in select conditions when the antiviral drug, amantadine, was added. Native mass spectrometry of AM2 in lipid nanodiscs with different lipids showed that lipids also affected the oligomeric states of AM2. Finally, nanodiscs uniquely enabled measurement of amantadine binding stoichiometries to AM2 in the intact lipid bilayer. These unexpected results reveal that AM2 forms a wider range of oligomeric states than previously thought possible, which provides new potential mechanisms of influenza pathology and pharmacology.

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