Marine Fish-Derived Lysophosphatidylcholine: Properties, Extraction, Quantification, and Brain Health Application.

Long-chain omega-3 fatty acids esterified in lysophosphatidylcholine (LPC-omega-3) are the most bioavailable omega-3 fatty acid form and are considered important for brain health. Lysophosphatidylcholine is a hydrolyzed phospholipid that is generated from the action of either phospholipase PLA1 or PLA2. There are two types of LPC; 1-LPC (where the omega-3 fatty acid at the sn-2 position is acylated) and 2-LPC (where the omega-3 fatty acid at the sn-1 position is acylated). The 2-LPC type is more highly bioavailable to the brain than the 1-LPC type. Given the biological and health aspects of LPC types, it is important to understand the structure, properties, extraction, quantification, functional role, and effect of the processing of LPC. This review examines various aspects involved in the extraction, characterization, and quantification of LPC. Further, the effects of processing methods on LPC and the potential biological roles of LPC in health and wellbeing are discussed. DHA-rich-LysoPLs, including LPC, can be enzymatically produced using lipases and phospholipases from wide microbial strains, and the highest yields were obtained by Lipozyme RM-IM®, Lipozyme TL-IM®, and Novozym 435®. Terrestrial-based phospholipids generally contain lower levels of long-chain omega-3 PUFAs, and therefore, they are considered less effective in providing the same health benefits as marine-based LPC. Processing (e.g., thermal, fermentation, and freezing) reduces the PL in fish. LPC containing omega-3 PUFA, mainly DHA (C22:6 omega-3) and eicosapentaenoic acid EPA (C20:5 omega-3) play important role in brain development and neuronal cell growth. Additionally, they have been implicated in supporting treatment programs for depression and Alzheimer's. These activities appear to be facilitated by the acute function of a major facilitator superfamily domain-containing protein 2 (Mfsd2a), expressed in BBB endothelium, as a chief transporter for LPC-DHA uptake to the brain. LPC-based delivery systems also provide the opportunity to improve the properties of some bioactive compounds during storage and absorption. Overall, LPCs have great potential for improving brain health, but their safety and potentially negative effects should also be taken into consideration.

Omega-3 phospholipids in Pacific blue mackerel (Scomber australasicus) processing by-products.

The present study investigated phospholipid content, fatty acid composition and the positional distribution (sn-1,3 and sn-2) of n-3 fatty acids in four blue mackerel processing by-products (head, skin, roe, and male gonad). Total lipid was extracted using hexane/ethanol (1:2) and the analyses were carried out using NMR and GC-FID techniques. On the basis of g wet tissue, blue mackerel roe was a better source of phospholipids (38.6 µmol), compared to head (9.89 µmol), skin (13.5 µmol), and male gonad (10.0 µmol). Total lipid extracted from roe was found to have a higher proportion of n-3 fatty acids (44.4%) including EPA (11.3%) and DHA (27.5%), compared to head (total n-3 = 36.6%; EPA, 9.08%: DHA, 21.9%), skin (total n-3 = 34.8%; EPA, 9.63%; DHA, 19.5%) and male gonad (total n-3 = 42.5%; EPA, 12.1%; DHA, 24.7%). The proportion of EPA in the sn-2 position was substantially higher in fish roe (12.6%) compared to the other by-products (head, 1.91%; skin, 2.22%; male gonad, 2.02%). However, the DPA and DHA content in the sn-2 position did not vary significantly among the various parts (p > 0.05). Phospholipid esterified n-3 fatty acids were higher in roe (55.5%) compared to head (40.9%), skin (21.8%) and male gonad (32%). The present study suggests that blue mackerel roe is the best source of marine n-3 phospholipids among the blue mackerel commercially produced by-products.

The Effect of the Supplementation of a Diet Low in Calcium and Phosphorus with Either Sheep Milk or Cow Milk on the Physical and Mechanical Characteristics of Bone using A Rat Model.

This study assessed the effect of cow milk (CM) and sheep milk (SM) consumption on the micro-structure, mechanical function, and mineral composition of rat femora in a male weanling rat model. Male weanling rats were fed a basal diet with a 50% reduction in calcium and phosphorus content (low Ca/P-diet) supplemented with either SM or CM. Rats were fed for 28 days, after which the femora were harvested and stored. The femora were analyzed by µ-CT, three-point bending, and inductively coupled plasma-mass spectrometry (ICP-MS). The addition of either milk to the low Ca/P-diet significantly increased (p < 0.05) trabecular bone volume, trabecular bone surface density, trabecular number, cortical bone volume, and maximum force, when compared to rats that consumed only the low Ca/P-diet. The consumption of either milk resulted in a significant decrease (p < 0.05) in trabecular pattern factor, and cortical bone surface to volume ratio when compared to rats that consumed only the low Ca/P-diet. The results were achieved with a lower consumption of SM compared to that of CM (p < 0.05). This work indicates that SM and CM can help overcome the effects on bone of a restriction in calcium and phosphorus intake.

PHNQ from Evechinus chloroticus Sea Urchin Supplemented with Calcium Promotes Mineralization in Saos-2 Human Bone Cell Line.

Polyhydroxylated naphthoquinones (PHNQs), known as spinochromes that can be extracted from sea urchins, are bioactive compounds reported to have medicinal properties and antioxidant activity. The MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) cell viability assay showed that pure echinochrome A exhibited a cytotoxic effect on Saos-2 cells in a dose-dependent manner within the test concentration range (15.625-65.5 µg/mL). The PHNQ extract from New Zealand sea urchin Evechinus chloroticus did not induce any cytotoxicity within the same concentration range after 21 days of incubation. Adding calcium chloride (CaCl2) with echinochrome A increased the number of viable cells, but when CaCl2 was added with the PHNQs, cell viability decreased. The effect of PHNQs extracted on mineralized nodule formation in Saos-2 cells was investigated using xylenol orange and von Kossa staining methods. Echinochrome A decreased the mineralized nodule formation significantly (p < 0.05), while nodule formation was not affected in the PHNQ treatment group. A significant (p < 0.05) increase in mineralization was observed in the presence of PHNQs (62.5 µg/mL) supplemented with 1.5 mM CaCl2. In conclusion, the results indicate that PHNQs have the potential to improve the formation of bone mineral phase in vitro, and future research in an animal model is warranted.

The Effect of Sheep and Cow Milk Supplementation of a Low Calcium Diet on the Distribution of Macro and Trace Minerals in the Organs of Weanling Rats.

The aim of this study was to investigate the effect of either sheep or cow milk supplementation to a low calcium and phosphorus diet on growth and organ mineral distribution in weanling rats. Rats were fed diets consisting of either a control chow, a 50% reduced calcium and phosphorous chow (low Ca/P), low Ca/P and sheep milk, or low Ca/P and cow milk diet for 28 days. Food intake of the rats, the growth rate of the rats, and the concentrations of minerals in the soft organs and serum were determined. Rats fed the low Ca/P diet alone had lower weight gain than rats consuming either of the milk-supplemented diets (p < 0.05). Both sheep milk and cow milk supplementation overcame the effects of consuming a diet restricted in calcium and phosphorus but the sheep milk was effective at a significantly lower level of milk intake (p < 0.05). Significant differences (p < 0.05) in essential and trace mineral concentrations due to milk type were observed in the kidney, spleen, and liver. For non-essential minerals, significant differences (p < 0.05), related to diet, were observed in all organs for arsenic, cesium, rubidium, and strontium concentrations.

Marine Waste Utilization as a Source of Functional and Health Compounds.

Consumer demand for convenience has led to large quantities of seafood being value-added processed before marketing, resulting in large amounts of marine by-products being generated by processing industries. Several bioconversion processes have been proposed to transform some of these by-products. In addition to their relatively low value conventional use as animal feed and fertilizers, several investigations have been reported that have demonstrated the potential to add value to viscera, heads, skins, fins, trimmings, and crab and shrimp shells by extraction of lipids, bioactive peptides, enzymes, and other functional proteins and chitin that can be used in food and pharmaceutical applications. This chapter is focused on reviewing the opportunities for utilization of these marine by-products. The chapter discusses the various products and bioactive compounds that can be obtained from seafood waste and describes various methods that can be used to produce these products with the aim of highlighting opportunities to add value to these marine waste streams.

Do Dairy Minerals Have a Positive Effect on Bone Health?

This review aims to examine the relationship between the consumption of dairy products, mineral absorption, and bone health, and critically evaluates the methods that have been used to investigate this relationship. As people live longer and have lives that are more active in modern societies, bone health is of concern due to the possibility for the increasing incidence of bone disorders, such as osteoporosis. It has been suggested that dairy products can play a key role in bone health due to their high levels of minerals. Whether the positive effect of dairy consumption on bone health is due solely to the concentration of minerals, the action of vitamins, proteins, and lipids present in dairy products, and complex interactions between different milk components remains to be determined. Assessment of how dairy products affect bone health is complex, with apparent contradictory conclusions being reported in the literature. To gain a better understanding of the effects that dairy products have on bone health, this review presents an evaluation of a combination of data obtained using a variety of methods. From those data, we surmise that the preferable approach to investigate the effects of milk on bone health is to obtain data from human, animal, and cell line testing. A combined approach will enable various aspects to be identified, including mechanisms and the assessment of holistic effects, which will enable the effects in the human situation to be ascertained.

Generation of bioactive peptide hydrolysates from cattle plasma using plant and fungal proteases.

Four protease preparations from plant and fungal sources (papain, bromelain, FP400 and FPII) were used to hydrolyse plasma which was separated from slaughterhouse cattle blood. The o-phthaldialdehyde assay was used to follow the release of TCA-soluble peptides over a 24h period. Hydrolysis profiles were displayed using SDS-PAGE. The in vitro antioxidant and antimicrobial activities of the hydrolysates were determined. The results showed that hydrolysates of cattle plasma generated with fungal protease FPII had higher antioxidant activities. Overall than hydrolysates generated with papain, bromelain and FP400. None of the hydrolysates demonstrated antimicrobial activity. The FPII peptide hydrolysate was fractionated using gel permeation chromatography, OFFGEL isoelectric focusing and RP-HPLC. The RP-HPLC fraction with highest antioxidant activity contained 15 novel peptide sequences. The use of protease FPII to hydrolyse cattle plasma resulted in a hydrolysate with high antioxidant properties and unique peptide sequences.

Production of bioactive peptide hydrolysates from deer, sheep, pig and cattle red blood cell fractions using plant and fungal protease preparations.

Protease preparations from plant (papain and bromelain) and fungal (FP400 and FPII) sources were used to hydrolyze the red blood cell fractions (RBCFs) separated from deer, sheep, pig, and cattle abattoir-sourced blood. After 1, 2, 4 and 24h of hydrolysis, the antioxidant and antibacterial activities of the peptide hydrolysates obtained were investigated. The increase in trichloroacetic acid-soluble peptides over the hydrolysis period was examined using the o-phthaldialdehyde (OPA) assay and the hydrolysis profiles were illustrated using SDS-PAGE. Papain generated RBCF hydrolysates exhibited higher ferric reducing antioxidant power (FRAP) and oxygen radical absorbance capacity (ORAC) compared to those generated with bromelain, FP400 and FPII. At certain concentrations, 24h hydrolysates of RBCF using FP400 and FPII were able to inhibit the growth of Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa. The results indicated that the use of proteases from plant or fungal sources can produce animal blood hydrolysates with antioxidant and antimicrobial activities.

Composition and biological activities of slaughterhouse blood from red deer, sheep, pig and cattle.

BACKGROUND: Animal blood is a large-volume by-product of the meat industry. Besides blood meal fertiliser, blood is marketed for human consumption as a supplement. Minimal comparative work on slaughterhouse animal blood fractions has been carried out. In this study, slaughterhouse deer, sheep, pig and cattle blood parameters were compared. Some blood constituents were determined. Fractionated blood was assessed for antioxidant activity (2,2-diphenyl-1-picrylhydrazyl radical scavenging, oxygen radical scavenging capacity and ferric reducing antioxidant power). Angiotensin converting enzyme (ACE) inhibitory activity and antimicrobial activity were also assessed. RESULTS: Serum iron ranged from 35.3 ± 0.6 µmol L(-1) in cattle to 16.3 ± 3.1 µmol L(-1) in deer. Cattle had the highest total plasma proteins (81.7 ± 1.5 g L(-1)). While the plasma fractions contained considerable antioxidant activity, the red blood cell fractions of all four animal species contained higher antioxidant activity (P < 0.05). Negligible levels of ACE inhibitory activity were found for all animal blood fractions. Antimicrobial activity was detected towards Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa with sheep white blood cells from which a crude neutrophil extract was obtained which demonstrated concentration-dependent inhibitory effects on the growth rates of these bacterial strains. CONCLUSION: Fractionated animal blood obtained from local slaughterhouses contains native proteins that possess antioxidant activity and antimicrobial activity.

Production of bioactive peptide hydrolysates from deer, sheep and pig plasma using plant and fungal protease preparations.

Plasma separated from deer, sheep and pig blood, obtained from abattoirs, was hydrolysed using protease preparations from plant (papain and bromelain) and fungal (FP400 and FPII) sources. Antioxidant and antimicrobial activities of the peptide hydrolysates obtained after 1, 2, 4 and 24h of hydrolysis, were investigated. The release of trichloroacetic acid-soluble peptides over the hydrolysis period was monitored using the o-phthaldialdehyde (OPA) assay, while the hydrolysis profiles were visualised using SDS-PAGE. The major plasma proteins in the animal plasmas were identified using MALDI-TOF-TOF MS. Hydrolysates of plasma generated with fungal proteases exhibited higher DPPH radical-scavenging, oxygen radical-scavenging capacity (ORAC) and ferric reducing antioxidant power (FRAP) than those generated with plant proteases for all three animal plasmas. No antimicrobial activity was detected in the hydrolysates. The results indicated that proteolytic hydrolysis of animal blood plasmas, using fungal protease preparations in particular, produces hydrolysates with high antioxidant properties.

Effects of L- and iso-ascorbic acid on meat protein hydrolyzing activity of four commercial plant and three microbial protease preparations.

The present study investigated the effects of both l- and iso-ascorbic acid (AA) on the activity of four plant proteases (papain, bromelain, actinidin and zingibain) and three microbial proteases (Bacterial Protease G, Fungal 31,000 and Fungal 60,000) preparations using fluorescent-labelled casein, meat myofibrillar and connective tissue extracts to explore their effects on meat structure components upon treatment with individual proteases. While l-AA in the range 0.8-3.2mM inhibited the activity of papain, bromelain and zingibain, iso-AA acted as an inhibitor of papain but as an activator of zingibain and had no significant effect on bromelain. Both AA isoforms acted as an activator of the actinidin protease and the concentration of AA isoforms appeared to affect the level of activation of the protease. The effect of the two AA isoforms on collagen and myofibrillar protein hydrolyzing activity varied depending on the concentration of the two AA isoforms. The results indicate the ability to up and down regulate the activity of the investigated proteases by using an appropriate concentration of the AA isoform.