Encapsulated Salmon Polar Lipids Modestly Reduce Postprandial Platelet Sensitivity to PAF and Thrombin in Healthy Adults: A Pilot Study

 The postprandial effects of a novel food-grade extracted salmon polar lipids (SPL) supplement against platelet-activating factor (PAF) and thrombin-induced platelet aggregation in a human pilot study was evaluated. This study was double-blinded, crossover, and placebo controlled in design. Five healthy volunteers completed 4 h time-course trials on 5 separate days. Blood was drawn at baseline before subjects consumed a standardized breakfast with one of the following treatments: a low-dose (0.25 g) or a high-dose (0.5 g) of SPL encapsulated within a stomach resistant capsule (LDSR and HDSR respectively) or a non-resistant capsule (LDSNR and HDSNR, respectively), or placebo capsules containing food-grade glycerin (0.5 g; Placebo). Blood was analyzed at 1 h intervals for 4 h. Among the treatments tested, the high-dose non-resistant capsule (HDSNR) produced the clearest postprandial effects, inducing significant increases in EC50 (reduced platelet sensitivity) for PAF at 2–3 h and for thrombin at 3–4 h post-ingestion. Smaller or delayed effects were observed with the high-dose resistant capsule (HDSR), while low-dose formulations produced minimal changes. Postprandial plasma glucose, lipid profile (TC, HDL-C, LDL-C, TG), fibrinogen, prothrombin time, and activated partial thromboplastin time remained unaffected across all trials. This pilot study provides the first in vivo indication that SPL supplementation may modestly attenuate platelet responsiveness to both PAF and thrombin without altering standard haemostatic or metabolic biomarkers. Larger controlled studies are warranted to confirm these preliminary findings and define optimal dosing and formulation strategies.

[source]

New publication from our group: Oil from Mullet Roe Byproducts: Effect of Oil Extraction Method on Human Erythrocytes and Platelets

 

 

Tsamesidis I, Tzika P, Georgiou D, Charisis A, Hans S, Lordan R, Zabetakis I, Kalogianni EP (2024) Oil

from Mullet Roe Byproducts: Effect of Oil Extraction Method on Human Erythrocytes and Platelets.

Foods 13(1) 79. https://doi.org/10.3390/foods13010079

Abstract

Background: The valorization of byproducts to obtain high nutritional value foods is of utmost importance for our planet where the population is booming. Among these products are oils rich in ω-3 fatty acids produced from fishery byproducts. Recently, mullet roe oil from roe byproducts was produced that was rich in the ω-3 fatty acids eicosatetraenoic acid (EPA) and docosahexaenoic acid (DHA). Oils are customarily characterized for their composition and degree of oxidation but little is known of their biological effects, especially the effect of the extraction method. 

 

Methods: The purpose of this study was to evaluate the effects of freshly extracted mullet roe oil from mullet roe byproducts and the effect of the extraction method on human red blood cells (hRBCs) and platelets. To this end, the hemocompatibility (cytotoxicity), oxidative effects, and erythrocyte membrane changes were examined after 1 and 24 h of incubation. Antiplatelet effects were also assessed in vitro.  

 

Results: The expeller press oil extraction method and alcalase-assisted extraction produced the most biocompatible oils, as shown by hemocompatibility measurements and the absence of erythrocyte membrane alterations. Solvent extracts and protease-assisted extraction oils resulted in the rupture of red blood cells at different examined dilutions, creating hemolysis.  

 

Conclusions: It seems that the proper functioning of oil–erythrocyte interactions cannot be explained solely by ROS. Further investigations combining chemical analysis with oil–cell interactions could be used as an input to design high nutritional value oils using green extraction technologies. All samples exhibited promising antiplatelet and antiblood clotting effects in vitro.

 

Keywords:

hemocompatibility; mullet roe; cytotoxicity; marine oil; reactive oxygen species; oil extraction; antiplatelet therapeutics

 

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This is the first publication stemmed from our collaboration with the research group of Prof Eleni P Kalogianni in the International Hellenic University in Greece.

Our publication is focusing on the valorisation of mullet roe oil and the bioactivities of extracts obrained from this byproduct. We show a promising way to valorise this oil. Our results should be useful for the aquaculture sector working on mullet farming.

Platelet activation: a promoter for psoriasis and its comorbidity, cardiovascular disease

Abstract

Psoriasis is a chronic inflammatory skin disease with a prevalence of 0.14% to 1.99%. The underlying pathology is mainly driven by the abnormal immune responses including activation of Th1, Th17, Th22 cells and secretion of cytokines. Patients with psoriasis are more likely to develop cardiovascular disease (CVD) which has been well recognized as a comorbidity of psoriasis. As mediators of hemostasis and thromboinflammation, platelets play an important part in CVD. However, less is known about their pathophysiological contribution to psoriasis and psoriasis-associated CVD. A comprehensive understanding of the role of platelet activation in psoriasis might pave the path for more accurate prediction of cardiovascular (CV) risk and provide new strategies for psoriasis management, which alleviates the increased CV burden associated with psoriasis. Here we review the available evidence about the biomarkers and mechanisms of platelet activation in psoriasis and the role of platelet activation in intriguing the common comorbidity, CVD. We further discussed the implications and efficacy of antiplatelet therapies in the treatment of psoriasis and prevention of psoriasis-associated CVD.

This paper is very important in helping us understanding the underlying biochemical phenomenon that causes inflammation and hence a number of chronic diseases.

I have a particular interest in skin diseases as I suffer from vitiligo and two friends of mine have psoriasis.

In this paper, the authors give us a novel insight linking (our work on) platelet activation to the development of CVD and psoriasis (see references 91 and 92)

A must read paper for all who are active in this field of research!

Ioannis   

New Book Chapter: Investigation of Platelet Aggregation in Atherosclerosis

 

A new book on Atherosclerosis has just been published where we have contributed a chapter.

The title of the book is

Atherosclerosis: Methods and Protocols

edited by Dipak Ramji and

our chapter is

Investigation of Platelet Aggregation in Atherosclerosis

Platelet activation and aggregation is implicated in all stages of inflammation-related atherosclerosis from the initial steps of endothelial dysfunction and plaque formation, to plaque rupture and atherothrombotic events, such as acute coronary syndrome, myocardial infarction, and ischemic incidences. Platelet aggregometry assays are the mainstream for evaluating and monitoring platelet reactivity in such conditions and for the investigation of prophylactic and therapeutic approaches. The most established methodology is light transmittance aggregometry (LTA). Here we describe the appropriate preparation of platelet suspensions from human blood and the methodology of LTA-based assays that is used for basic and clinical research for monitoring and evaluating the activities of several thrombotic mediators, as well as determining the dose efficacy and safety of several pharmaceutical and nutraceutical compounds intended for therapeutic and prophylactic interventions for atherosclerosis.

Key words

Atherosclerosis Inflammation Thrombosis Platelet aggregation Light transmission aggregometry Platelet-activating factor Thrombin ADP Collagen Antiplatelet 

This chapter is dedicated to Prof. Constantinos A. Demopoulos, Εmeritus Professor at the University of Athens, Greece, our valued teacher, colleague, and friend.

new paper from our group: Anti-Platelet Properties of Apple Must/Skin Yeasts and of Their Fermented Apple Cider Products

 Anti-Platelet Properties of Apple Must/Skin Yeasts and of Their Fermented Apple Cider Products

 

 

Abstract

Alcoholic beverages like apple cider are considered functional beverages with several health benefits, when consumed in moderation, which are mainly attributed to their microbiota and the plethora of their bioactive compounds. Among them, bio-functional polar lipids (PL) have recently been found in apple cider, which despite low quantities, have exhibited strong anti-inflammatory and anti-platelet properties, while fermentation seems to affect the functionality of apple cider’s PL bioactives. The aim of the present study was to elaborate yeast strains isolated from the complex mixtures of apple surface and must yeasts for evaluating their effects on the anti-platelet functional properties of PL bioactives from their final fermented apple cider products. First, bio-functional PL were extracted and separated from the biomass of the different isolated apple surface/must yeast strains, and were further assessed for their anti-platelet potency against human platelet aggregation induced by the potent inflammatory and thrombotic mediator platelet-activating factor (PAF), or by a classic platelet agonist like adenosine diphopshate (ADP). Novel functional apple ciders were then produced from the fermentation of apple juice by elaborating the most bioactive and resilient yeast strains isolated from the apple must with optimum fermentation properties. PL bioactives extracted from these novel apple cider products were also further assessed for their anti-platelet properties against both the PAF and ADP pathways of human platelet aggregation. These novel cider products were found to contain PL bioactives with lower IC50 values (~40 μg) and thus increased anti-platelet potency against platelet aggregation induced by PAF and ADP. GC-MS analysis of the PL bioactives extracted from these novel apple ciders showed that apple cider PL bioactives are rich in monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA), such as the omega-6 linoleic acid (LA) and the omega-3 alpha linolenic acid (ALA), with favorably lower levels for their omega-6/omega-3 PUFA ratio, which further support the observed strong anti-platelet properties putative anti-inflammatory potency for the apple cider PL bioactives. However, further studies are needed in order to elucidate and fully characterize the apple yeast strains that can be utilized for increasing the anti-inflammatory, anti-platelet and cardioprotective functional properties of their fermented apple cider products. View Full-Text

New Paper from our group: Antithrombotic and antiplatelet activity of an organometallic rhodium(I) complex incorporating a substituted thieno‐[2,3‐d]‐pyrimidine ligand: Synthesis, structural characterization, and molecular docking calculations

 New paper just published in Appl. Organomet. Chem.

Abstract

The antiplatelet and antithrombotic activity of a novel organometallic rhodium(I) complex of the formula [Rh(cod)Cl(tpc)] (1) (cod = cis‐1,5‐cyclooctadiene; tpc = methyl 2‐amino‐4‐(diethylamino)‐thieno‐[2,3‐d]‐pyrimidine‐6‐carboxylate) was investigated. Complex 1 was easily synthesized by a one‐pot, high‐yield reaction and was fully characterized by standard spectroscopic techniques including FT‐IR, UV–Vis, and NMR spectroscopy and by elemental analysis. The molecular structures of tpc and 1 were determined by single‐crystal X‐ray crystallography. Complex 1 displayed a slightly distorted square planar geometry and is the first crystallographically characterized example of a coordination compound bearing the ligand precursor tpc. Biological studies demonstrate that 1 displays strong antiplatelet and antithrombotic properties in vitro, by inhibiting both the aggregation of human and washed rabbit platelets induced by the potent inflammatory and thrombotic mediator, platelet‐activating factor (PAF) in the micromolar range. This is an approach of continuous interest in the field. Molecular docking calculations suggest that 1 can fit at the ligand‐binding site of the PAF receptor (PAFR) and thus block PAF thrombotic activities, through an antagonistic effect on the PAF/PAFR‐related pathway, which is in accord with the experimental findings. Complex 1 constitutes an interesting example of a metal‐based PAF inhibitor with promising antiplatelet, antithrombotic, and anti‐inflammatory activity, because PAF is the most potent inflammatory lipid mediator. This is also supported by the fact that 1 is an inhibitor of other inflammatory and thrombotic mediators like thrombin, along with well‐established platelet agonists like ADP and collagen.

New paper from our group

 

Platelet activation and prothrombotic mediators at the nexus of inflammation and atherosclerosis: Potential role of antiplatelet agents

 Blood Reviews

Available online 21 April 2020, 100694

Abstract

Platelets are central to inflammation-related manifestations of cardiovascular diseases (CVD) such as atherosclerosis. Platelet-activating factor (PAF), thrombin, thromboxane A₂ (TxA₂), and adenosine diphosphate (ADP) are some of the key agonists of platelet activation that are at the intersection between a plethora of inflammatory pathways that modulate pro-inflammatory and coagulation processes. The aim of this article is to review the role of platelets and the relationship between their structure, function, and the interactions of their constituents in systemic inflammation and atherosclerosis. Antiplatelet therapies are discussed with a view to primary prevention of CVD by the clinical reduction of platelet reactivity and inflammation. Current antiplatelet therapies are effective in reducing cardiovascular risk but increase bleeding risk. Novel therapeutic antiplatelet approaches beyond current pharmacological modalities that do not increase the risk of bleeding require further investigation. There is potential for specifically designed nutraceuticals that may become safer alternatives to pharmacological antiplatelet agents for the primary prevention of CVD but there is serious concern over their efficacy and regulation, which requires considerably more research.

 

new paper from our group

Total, Neutral, and Polar Lipids of Brewing Ingredients, By-Products and Beer: Evaluation of Antithrombotic Activities 

Ronan Lordan ^*, Eoin O’Keeffe, Alexandros Tsoupras and Ioannis Zabetakis

Department of Biological Sciences, University of Limerick, Limerick V94 T9PX, Ireland

^*

Author to whom correspondence should be addressed.

Foods 2019, 8(5), 171; https://doi.org/10.3390/foods8050171 (registering DOI)

Received: 24 April 2019 / Revised: 14 May 2019 / Accepted: 19 May 2019 / Published: 20 May 2019 

 

the full text is here.

New paper of our team

 

 

Platelet aggregometry assay for evaluating the effects of platelet agonists and antiplatelet compounds on platelet function in vitro

 AlexandrosTsoupras IoannisZabetakis RonanLordan

Department of Biological Sciences, University of Limerick, Limerick, V94 T9PX, Ireland

 

MethodsX

Volume 6, 2019, Pages 63-70

 

Abstract

Platelet aggregometry assays are generally used for the analysis of platelet function but can also be adapted for further research and therapy focused applications. This method describes the procedures for the preparation of human platelet-rich plasma (PRP) and platelet-poor plasma (PPP) for the assessment of human platelet aggregation induced by agonists such as platelet-activating factor (PAF), thrombin, collagen, adenosine diphosphate (ADP), arachidonic acid, etc.

•

This method can be applied in vitro to evaluate the aggregatory effects of these agonists and to assess the antiaggregatory effects of several bioactive antiplatelet agents (compounds of natural or pharmacological origin) in human PRP.

•

This versatile method can be used in both basic and clinical research for the assessment of platelet aggregation (a major cardiovascular risk factor), platelet agonists, and inhibitors, in physiological or pathological conditions.

•

This method can be adapted to assess platelet activity in postprandial and intervention studies ex vivo.

 

Structural Elucidation of Irish Organic Farmed Salmon (Salmo salar) Polar Lipids with Antithrombotic Activities

just published in Marine Drugs

Alexandros Tsoupras ¹, Ronan Lordan ¹, Martina Demuru ^1,2, Katie Shiels ³, Sushanta Kumar Saha ³, Constantina Nasopoulou ^4,* and Ioannis Zabetakis ¹

¹

Department of Biological Sciences, University of Limerick, V94 T9PX Limerick, Ireland

²

Department of Life and Environmental Sciences, University of Cagliari, via Ospedale 72, 09124 Cagliari, Italy

³

Shannon Applied Biotechnology Centre, Limerick Institute of Technology, Moylish Park, V94 E8YF Limerick, Ireland

⁴

Department of Food Science and Nutrition, School of the Environment, University of the Aegean, GR 81400 Myrina, Lemnos, Greece

^*

Author to whom correspondence should be addressed.

Received: 17 April 2018 / Revised: 16 May 2018 / Accepted: 22 May 2018 / Published: 23 May 2018

(This article belongs to the Special Issue Marine-derived Bioactive Peptides and Lipids Applied in Non-communicable Diseases)

View Full-Text   |   Download PDF [3602 KB, uploaded 23 May 2018]   |  

Browse Figures

 

 

Graphical Abstract

 

Abstract

While several marine polar lipids (PL) have exhibited cardioprotective properties through their effects on the platelet-activating factor (PAF) pathways, salmon PL have not been tested so far. In this study, the antithrombotic activities of salmon PL were assessed in human platelets and the structural characterisation of bioactive salmon PL was performed by GC-MS and LC-MS analyses. PL from fillets of Irish organic farmed salmon (Salmo salar) were extracted and separated into several lipid subclasses by thin-layer chromatography (TLC), while their fatty acid profile was fully characterised by GC-MS. Salmon total lipids (TL), total neutral lipids (TNL), total polar lipids (TPL), and each PL subclass obtained by TLC were further assessed for their in vitro effects towards PAF-induced and thrombin-induced platelet aggregation in human platelets. Salmon PL exhibited antithrombotic effects on human platelet aggregation, mostly through their strong inhibitory effects against the PAF pathway with IC₅₀ values comparable to other marine PL, but with lower effects towards the thrombin pathway. PL fractions corresponding to phosphatidylcholine and phosphatidylethanolamine derivatives exhibited the most potent anti-PAF effects, while LC-MS analysis putatively elucidated their structure/function relationship. Several diacyl-PC/PE and alkyl-acyl-PC/PE species containing mostly docosahexaenoic acid at their sn-2 glycerol-backbone may be responsible for the bioactivity. The data presented suggests that salmon contains PL with strong antithrombotic bioactivities.

 

 View Full-Text

 

Keywords: salmon; polar lipids; platelet aggregation; platelet-activating factor (PAF); thrombin; antithrombotic; LC-MS; GC-MS; phosphatidylcholine; phosphatidylethanolamine; polyunsaturated fatty acids (PUFA); docosahexaenoic acid (DHA)

Evaluation of olive pomace in the production of novel broilers with enhanced in vitro antithrombotic properties

Evaluation of olive pomace in the production of novel broilers with enhanced in vitro antithrombotic properties^†

^{C.N, K.L and I.Z.}

• ^†This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. Please cite this article as doi: [10.1002/ejlt.201700290].

Abstract

Several attempts have been made not only to improve nutritional value of broilers but also to attenuate dependence on raw materials such as corn in compounded broilers feed. Therefore the aim of this study was to evaluate the impact of diets enriched with olive pomace (OP) on Ross 308 broilers growth performance, sensory characteristics and nutritional value in terms of cardioprotection. Broilers were fed four experimental diets containing 0 % OP (control group), 2.5 % OP (group A), 5.0 % OP (group B) and 7.5 % OP (group C). The obtained broiler meat samples were evaluated for their lipid and phenol content and their in vitro antithrombotic properties according to biological assay in human platelets. Groups B and C exhibited significantly increased (P < 0.05) growth rate compared to the ones of control group. Additionally group B exhibited significantly more potent (P < 0.05) in vitro antithrombotic properties (EC₅₀ = 10.5±0.92) compared to the ones of control group (EC₅₀ = 420±21.3). Grilled broiler meat of group B was found to have acceptable sensory properties. The overal conclusion of this paper is the potential use of OP in compounded broilers feed in the production of functional broilers meat.

Practical applications: The objective of this research is to assess the use of olive industry by-products as functional feed ingredients. For this purpose, broilers were fed experimental diets containing olive pomace (OP). Our results suggest that OP can be used in broiler feed to produce functional broilers meat with increased in vitro antithrombotic properties. These scientific data could have considerable practical value towards the valorisation of OP and increasing the sustainable production of functional broiler meat and therefore the overall food security.

New paper: Evaluation of sensory and in vitro anti-thrombotic properties of traditional Greek yogurts derived from different types of milk

In vitro biological activities of total lipids (TL) and total polar lipids (TPL) of cow, ewe and goat yogurts towards washed rabbit platelet aggregation, expressed in micrograms of TL and TPL. Washed rabbit platelet concentration was approximately 500,000 platelets μL ⁻¹. The final concentration of PAF in the cuvette was 2.5 × 10⁻¹¹ M. The experimental analyses were carried out in triplicate (n = 3).

 

Our latest paper on yogurts has just been published in Heliyon.

Evaluation of sensory and in vitro anti-thrombotic properties of traditional Greek yogurts derived from different types of milk

Abstract

Given that fermented dairy products exhibit high bioactivities against cardiovascular diseases (CVDs), the anti-thrombotic properties, fatty acid profiles and sensory properties of cow, goat and ewe derived Greek yogurts have been assessed and compared. The total lipids (TL), total polar lipids (TPL), total neutral lipids (TNL) were obtained and the polar lipid fractions were further separated by thin layer chromatography (TLC). These lipid samples (TL, TPL and TLC fractions) were subsequently assessed for their biological activity against atherosclerosis based on the in vitro inhibition of Platelet Activating Factor (PAF)-induced platelet aggregation. The fatty acid compositions of all yogurts were analyzed by Gas Chromatography with flame ionization detector (GC-FID). Goat yogurt lipids have been found to exert more potent inhibitory activity (i.e. lower IC₅₀ values in both TL and TPL samples) in contrast to the corresponding fractions of cow and ewe yogurts. The observed sensory data indicates that ewe yogurt was the most palatable of all three Greek yogurts.

 

You can download it here.