scholarly journals Bioactive Lipids of Marine Microalga Chlorococcum sp. SABC 012504 with Anti-Inflammatory and Anti-thrombotic Activities

Marine Drugs ◽  
2021 ◽  
Vol 19 (1) ◽  
pp. 28
Author(s):  
Katie Shiels ◽  
Alexandros Tsoupras ◽  
Ronan Lordan ◽  
Constantina Nasopoulou ◽  
Ioannis Zabetakis ◽  
...  
Keyword(s):  
Essential Fatty Acids ◽  
Lipid Fraction ◽  
Platelet Activating Factor ◽  
Human Platelets ◽  
Bioactive Molecules ◽  
Bioactive Lipids ◽  
Alternative Source ◽  
Lipid Fractions ◽  
Anti Inflammatory

Microalgae are at the start of the food chain, and many are known producers of a significant amount of lipids with essential fatty acids. However, the bioactivity of microalgal lipids for anti-inflammatory and antithrombotic activities have rarely been investigated. Therefore, for a sustainable source of the above bioactive lipids, the present study was undertaken. The total lipids of microalga Chlorococcum sp., isolated from the Irish coast, were fractionated into neutral-, glyco-, and phospho-lipids, and were tested in vitro for their anti-inflammatory and antithrombotic activities. All tested lipid fractions showed strong anti-platelet-activating factor (PAF) and antithrombin activities in human platelets (half maximal inhibitory concentration (IC50) values ranging ~25–200 μg of lipid) with the highest activities in glyco- and phospho-lipid fractions. The structural analysis of the bioactive lipid fraction-2 revealed the presence of specific sulfoquinovosyl diacylglycerols (SQDG) bioactive molecules and the HexCer-t36:2 (t18:1/18:1 and 18:2/18:0) cerebrosides with a phytosphingosine (4-hydrosphinganine) base, while fraction-3 contained bioactive phosphatidylcholine (PC) and phosphatidylethanolamine (PE) molecules. These novel bioactive lipids of Chlorococcum sp. with putative health benefits may indicate that marine microalgae can be a sustainable alternative source for bioactive lipids production for food supplements and nutraceutical applications. However, further studies are required towards the commercial technology pathways development and biosafety analysis for the use of the microalga.

scholarly journals In Vitro Antithrombotic Properties of Salmon (Salmo salar) Phospholipids in a Novel Food-Grade Extract

Marine Drugs ◽  
2019 ◽  
Vol 17 (1) ◽  
pp. 62 ◽  
Author(s):  
Alexandros Tsoupras ◽  
Ronan Lordan ◽  
Katie Shiels ◽  
Sushanta Saha ◽  
Constantina Nasopoulou ◽  
...  
Keyword(s):  
Platelet Aggregation ◽  
Salmo Salar ◽  
Platelet Activating Factor ◽  
Human Platelets ◽  
Food Grade ◽  
Farmed Salmon ◽  
Lipid Fractions ◽  
Antithrombotic Effects ◽  
Layer Chromatography

Marine and salmon polar lipids (PLs) extracted by conventional extractions with non-food-grade solvents (CE-salmon-PLs) possess antithrombotic bioactivities against platelet-activating factor (PAF) and thrombin. Similar effects of food-grade-extracted (FGE) marine PLs have not yet been reported. In this study, food-grade solvents were used to extract PLs from Irish organic farmed salmon (Salmo salar) fillets (FGE-salmon-PLs), while their antithrombotic bioactivities were assessed in human platelets induced by platelet aggregation agonists (PAF/thrombin). FGE-salmon-PLs were further separated by thin layer chromatography (TLC) into lipid subclasses, and the antithrombotic bioactivities of each subclass were also assessed. LC-MS was utilized to elucidate the structure-activity relationships. FGE-salmon-PLs strongly inhibited PAF-induced platelet aggregation, while their relevant anti-thrombin effects were at least three times more potent than the previously reported activities of CE-salmon-PLs. TLC-derived lipid fractions corresponding to phosphatidylcholines (PC) and phosphatidylethanolamines (PE) were the most bioactive lipid subclasses obtained, especially against thrombin. Their LC-MS analysis elucidated that they are diacyl- or alkyl-acyl- PC and PE moieties baring ω3 polyunsaturated fatty acids (PUFA) at their sn-2 position, such as eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA). Our results concerning the potent antithrombotic effects of FGE-salmon-PLs against both PAF and thrombin pathways strongly suggest that such food-grade extracts are putative candidates for the development of novel cardioprotective supplements and nutraceuticals.

Continuous in vitro culture of Babesia divergens in a serum-free medium

Parasitology ◽  
1997 ◽  
Vol 115 (1) ◽  
pp. 81-89 ◽  
Author(s):  
N. GRANDE ◽  
E. PRECIGOUT ◽  
M. L. ANCELIN ◽  
K. MOUBRI ◽  
B. CARCY ◽  
...  
Keyword(s):  
Human Serum ◽  
Reduced Glutathione ◽  
Lipid Fraction ◽  
Basal Medium ◽  
Defined Medium ◽  
Specific Lipids ◽  
Serum Free ◽  
Babesia Divergens ◽  
Lipid Fractions

Babesia divergens was cultivated in RPMI 1640 (25 mM HEPES) supplemented with 10% human serum (RPMI-10% HS) with a high percentage of parasitized erythrocytes (PPE) ([ges ]40%). Standardization of in vitro tests, purification of exoantigens, biochemical studies and the safety of the culture handler motivated the development of a serum-free defined medium. Removal of serum greatly reduced the PPE but, after a period of adaptation, the culture was continuous and the parasite was able to develop a 3% routine PPE. Addition of vitamins or reduced glutathione in basal medium (RPMI) did not improve the PPE. The supplementation of basal medium with lipidic carrier (Albumax I or bovine serum albumin–Cohn's fraction V) promoted the growth of B. divergens with high PPE (>30%) close to those obtained in RPMI–10% HS. Neither protein nor lipid fractions alone were able to restore the growth of B. divergens. Nevertheless, the whole lipid fraction from serum or Albumax I added to delipidated albumin partially restored the growth (7% PPE), indicating that the presentation of specific lipids by a carrier is crucial for the parasite. All the data indicate that Albumax I can replace human serum offering the advantages of safety, standardization for chemosensitivity tests, and exoantigen purification.

scholarly journals In-vitro Assessment of Biological Properties of Lipids Isolated from Gonads of Stomopneustes variolaris

2020 ◽  
Vol 2 (1) ◽  
pp. 66
Keyword(s):  
Biological Effects ◽  
Lipid Fraction ◽  
Radical Scavenging ◽  
Biological Properties ◽  
Total Phenolic ◽  
Klebsiella Pneumonia ◽  
Spectroscopic Techniques ◽  
Lipid Fractions ◽  
Anti Fungal

Lipid fractions of gonads present in sea urchins serves as a source of bioactive agents with potent pharmaceutical properties. The present study reports the in-vitro biological effects of lipids isolated from gonads of sea urchin: Stomopneustes variolaris collected from the East coast of India. The extracted lipids were characterized by spectroscopic techniques such as GCMS and FTIR and tested for in-vitro biological effects. GCMS analysis of the lipid extract detected high levels of hexa triacontane (17.023 %), tetratetracontane (15.913%), and octacosane (15.628%) and low concentrations of oleic acid (2.206%) and sulfurous acid, pentadecy 2-propyl ester (1.744%). FTIR analysis identified rich composition of functional groups present in the lipids such as 3418.93 cm-1 (hydroxyl), 2921.08cm-1 and 2854.81 cm-1 (alkane), 2660.69 cm-1 (carboxylic acid), 1596.11 cm-1 (amine), 1291.76 cm-1 (aromatic amine). The lipid fraction evaluated by agar diffusion assay measured in terms of zone of inhibition showed bactericidal effects against gram-positive bacteria: Streptococcus aureus (30 mm); Pseudomonas aeruginosa (28.5 mm) and gram-negative bacteria: Escherichia coli (29.5 mm); Klebsiella pneumonia (27.5 mm) and Vibrio cholera (28 mm) respectively. The lipid fraction also showed an effective anti-fungal effect against C.albicans (25 mm). Further, the lipid fractions showed good radical scavenging effect against total phenolic, flavonoid content (15.12 mg GAE/g and 32.72 mg QE/g), and hydrogen peroxide radicals (IC50- 48.28mg/ml) confirming its anti-oxidant potential. Based on the observed results, it was identified that the lipid fraction of gonads of Stomopneustes variolaris demonstrated various biological effects such as bactericidal, anti-fungal and radical scavenging activities which could have a great scope in the formulation of biopharmaceutical agents.

scholarly journals In vitro health beneficial activities of Pumpkin seeds from Cucurbita moschata cultivated in Lemnos

2015 ◽  
Vol 4 (2) ◽  
Author(s):  
Danai Sakka ◽  
Haralabos C. Karantonis
Keyword(s):  
Platelet Activating Factor ◽  
Health Improvement ◽  
Cucurbita Moschata ◽  
Pumpkin Seed ◽  
Anti Oxidant ◽  
Pumpkin Seeds ◽  
Anti Inflammatory ◽  
Locally Grown ◽  
Seed Extracts

Pumpkin seeds are commonly consumed in Greece. Although Cucurbita moschata is locally grown in Lemnos and is traditionally used in pumpkin pies, the seeds are currently discarded after consumption of the fruit flesh. The aim of the present study was to investigate the nutritional functionality of pumpkin seeds from Cucurbita moschata grown in Lemnos.Cucurbita moschatas’ seeds, raw or roasted, were appropriately extracted and the results are presented for raw versus (vs) roasted seed extracts. The phenolic content was expressed as μg gallic acid/g of seeds according to Folin-Ciocalteau assay (370.3 ± 19.1 vs 551.0 ± 22.0). Antioxidant capacity was expressed as equivalent amount for 50% scavenging in mg of seeds for DPPH (50.03 ± 5.91 vs 25.82 ± 6.77) and ABTS (17.85 ± 0.77 vs 12.77 ± 0.76) assays, and as μmol of trolox/g of seeds for FRAP (1.19 ± 0.05 vs 2.50 ± 0.23) and CUPRAC (2.13 ±0.11 vs 3.25 ± 0.06) assays. Anti-inflammatory/anti-thrombotic and anti-diabetic activities were expressed as mg of seeds for 50% inhibition of platelet activating factor (0.62 vs 0.15) and as μg of seeds for 25% inhibition of alpha-glycosidase (40.0 vs 61.0) activities respectively. Moreover, anti-atherogenic activity was expressed as the % increase in lag time of human plasma oxidation (62.7 versus 163.2)Raw and roasted pumpkin seed extracts exert anti-oxidant, anti-thrombotic/anti-inflammatory, anti-atherogenic and antidiabetic activities. Cucurbita moschata seeds may represent a novel opportunity for development of functional foods, with a local interest in Lemnos that would contribute also to the regional public health improvement.

IS IT POSSIBLE TO IDENTIFY HUMAN PLATELETS IN VITRO AS BEING DESENSITIZED TO PLATELET ACTIVATING FACTOR (PAF-ACETHER,PAF) IN VIVO?

1987 ◽  
Author(s):  
Cs Perger ◽  
A von Felten
Keyword(s):  
Platelet Activating Factor ◽  
Test System ◽  
Human Platelets ◽  
Vacuum Filtration ◽  
Reversible Aggregation ◽  
The Individual ◽  
20 Nm ◽  
And Control

PAF is suggested to be of pathophysiological importance in a variety of diseases. Since platelets exhibit a reduced sensitivity to PAF after a contact with this agent, this behavior may be used as indicator of PAF released into the circulation. In extrinsic asthma, platelets show a diminished reaction to PAF after exposition of the patients to the antigen compared to their own platelets before exposition (Beer and von Felten, Adv. Inflamm. Res. 10:323,1986). We were therefore looking for a test system indicating directly whether platelets had been in contact with PAF.Preparation of PAF-desensitized platelets: Citrated PRP was placed in a cuvette of an aggregometer, and PAF was added in 10 portions at intervals of 10 sec (37oc, constant stirring) to a final concentration of 10 to 100 nM, depending on the individual sensitivity of each platelet preparation. Therby, only a minimal, completely reversible aggregation was registered without any release of serotonin (ST) or 3-thromboglobulin (BTG). Control platelets were pretreated with buffer instead of PAF. Both platelets preparations were kept at 37°C for 45 min. Whereas control platelets showed a secondary aggregation to PAF (5x conc. used for desensitization), PAF-pretreated piatelets were only reversibly aggregated.Sensitivity of PAF-desensitized and control platelets to other platelet agonists: No difference in aggregation, ST-or BTG-relea-se was observed after stimulation with several concentrations of ADP, collagen and arachidonate (p>0.05,n= 41).Binding of 3H-PAF to platelets: PAF-desensitized and control platelets were separated from plasma by filtration through sepharose CL-2B (Pharmacia) in hepes-buffered Tyrode’s solution. After incubation with 3H-PAF, platelets were washed on Whatman 934-AH filters (vacuum filtration). On desensitized and control platelets, we found 175±48 (mean±sd) and 231±70 3H-PAF molecules / platelet respectively after incubation with 5 nM ^h-PAF, 399±36 and 504±66 ^H-PAF molecules / platelet after incubation with 20 nM. In spite of a statistically significant reduction of PAF-binding after desensitization (p<0.01),the variability of PAF-binding between platelets of different individuals is too high to allow a discrimination of normal from PAF-desensitized platelets.

scholarly journals Calcium-dependent cysteine protease activity in the sera of patients with thrombotic thrombocytopenic purpura

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1683-1687 ◽  
Author(s):  
WG Murphy ◽  
JC Moore ◽  
JG Kelton
Keyword(s):  
Thrombotic Thrombocytopenic Purpura ◽  
Cysteine Protease ◽  
Platelet Activating Factor ◽  
Aminocaproic Acid ◽  
Human Platelets ◽  
Thrombocytopenic Purpura ◽  
Platelet Surface ◽  
Calcium Dependent

Abstract Plasma and serum from patients with thrombotic thrombocytopenic purpura (TTP) can cause activation and aggregation of normal human platelets in vitro. It is possible that this platelet-activating factor contributes to the disease. In this report we describe studies designed to identify the platelet-activating factor in TTP. Platelet activation by sera from 15 patients with TTP was inhibited by leupeptin, iodoacetamide, and antipain but not by phenylmethylsulphonylfluoride, epsilon-aminocaproic acid, soybean trypsin inhibitor, aprotinin, and D-phenylanyl-1-prolyl-1- arginine chloromethyl ketone. These studies suggested that the platelet- activating factor in TTP serum was a cysteine protease. We confirmed that a calcium-dependent cysteine protease (CDP) was present in the sera of each of the 15 patients when we used an assay based on the ability of CDP to proteolyse platelet membrane glycoprotein 1b (GP1b) and hence to abolish the ability of CDP-treated normal platelets to agglutinate in the presence of ristocetin and von Willebrand factor. This proteolytic activity was inhibited by EDTA, leupeptin, antipain, iodoacetamide, and by N-ethyl-maleamide (NEM) but not by the serine protease inhibitors. Activity was detected in 15 of 15 patients with TTP tested before therapy was begun. In contrast, no activity was detected in the serum of any of five of the TTP patients tested in remission or in any of the sera from 36 patients with thrombocytopenia and 423 nonthrombocytopenic controls. To look for in vivo CDP activity in patients with TTP, we studied platelets from two patients with acute TTP (drawn into acid-citrate-dextrose, NEM, and leupeptin). These platelets showed a loss of GP1b from the platelet surface. Both patients were also studied in remission: GP1b on the platelet surface had returned to normal. These studies provide evidence that CDP is present in the sera of patients with TTP, that it is specific to this disease, and that is is active in vivo as well as in vitro. We postulate that a disorder of CDP homeostasis plays a major role in the pathophysiology of TTP.

scholarly journals Potential cardioprotective peptides generated in Spanish dry-cured ham

2019 ◽  
Vol 6 ◽  
Author(s):  
Marta Gallego ◽  
Leticia Mora ◽  
Fidel Toldrá
Keyword(s):  
Cardiovascular Health ◽  
Platelet Activating Factor ◽  
Developed Countries ◽  
Platelet Activating Factor Acetylhydrolase ◽  
Angiotensin I Converting Enzyme ◽  
Lipoxygenase Inhibition ◽  
Dry Cured Ham ◽  
Anti Inflammatory

Food-derived bioactive peptides are promising compounds for the prevention and treatment of cardiovascular diseases, the main cause of mortality in developed countries. The aim of this work was to determine the in vitro anti-inflammatory, antioxidant, and angiotensin I-converting enzyme (ACE-I) inhibitory activities of twenty-four peptides that were identified in Spanish dry-cured hams. For the first time, some peptides such as PSNPP, HCNKKYRSEM and FNMPLTIRITPGSKA showed anti-inflammatory activity expressed as platelet-activating factor-acetylhydrolase, autotaxin, and lipoxygenase inhibition. Peptides MDPKYR and TKYRVP were the strongest antioxidants, whereas GGVPGG, TKYRVP, and HCNKKYRSEM showed the highest ACE-I inhibitory activity. Additionally, several peptides such as KPVAAP, MDPKYR, TKYRVP, and HCNKKYRSEM presented more than one of the assayed activities, increasing their health-enhancing potential. More studies are needed to evaluate the bioavailability of such peptides and their in vivo effect. This would contribute to consider dry-cured ham as a source of peptides beneficial for cardiovascular health.

scholarly journals In vitro anti-inflammatory and anti-coagulant effects of antibiotics towards Platelet Activating Factor and thrombin

2011 ◽  
Vol 8 (1) ◽  
pp. 17 ◽  
Author(s):  
Alexandros B Tsoupras ◽  
Maria Chini ◽  
Nickolaos Tsogas ◽  
Athina Lioni ◽  
George Tsekes ◽  
...  
Keyword(s):  
Platelet Activating Factor ◽  
Anti Inflammatory

Nonsteroidal anti-inflammatory drugs in-vitro and in-vivo treatment and Multidrug Resistance Protein 4 expression in human platelets

2016 ◽  
Vol 76 ◽  
pp. 11-17 ◽  
Author(s):  
Flavia Temperilli ◽  
Manuela Di Franco ◽  
Isabella Massimi ◽  
Maria Luisa Guarino ◽  
Maria Paola Guzzo ◽  
...  
Keyword(s):  
Multidrug Resistance ◽  
Human Platelets ◽  
Multidrug Resistance Protein ◽  
Inflammatory Drugs ◽  
Resistance Protein ◽  
In Vivo Treatment ◽  
Anti Inflammatory
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