1-O-alkyl-glycerols from Squid Berryteuthis magister Reduce Inflammation and Modify Fatty Acid and Plasmalogen Metabolism in Asthma Associated with Obesity.

Asthma associated with obesity is considered the most severe phenotype and can be challenging to manage with standard medications. Marine-derived 1-O-alkyl-glycerols (AGs), as precursors for plasmalogen synthesis, have high biological activity, making them a promising substance for pharmacology. This study aimed to investigate the effect of AGs from squid Berryteuthis magister on lung function, fatty acid and plasmalogen levels, and cytokine and adipokine production in obese patients with asthma. The investigational trial included 19 patients with mild asthma associated with obesity who received 0.4 g of AGs daily for three months in addition to their standard treatment. The effects of AGs were evaluated at one and three months of treatment. The results of the study demonstrated that intake of AGs increased the FEV1 and FEV1/VC ratios, and significantly decreased the ACQ score in 17 of the 19 patients after three months of treatment. The intake of AGs increased concentration of plasmalogen and n-3 PUFA in plasma, and modified leptin/adiponectin production by adipose tissue. The supplementation of AGs decreased the plasma levels of inflammatory cytokines (TNF-α, IL-4, and IL-17a), and oxylipins (TXB2 and LTB4), suggesting an anti-inflammatory property of AGs. In conclusion, 1-O-alkyl-glycerols could be a promising dietary supplement for improving pulmonary function and reducing inflammation in obese asthma patients, and a natural source for plasmalogen synthesis. The study highlighted that the beneficial effects of AG consumption can be observed after one month of treatment, with gradual improvement after three months of supplementation.

[Squid fat as a promising source of polyunsaturated fatty acids].

Omega-3 polyunsaturated fatty acids (ω-3 PUFA) are substances that play an important role in human metabolism. They are essential nutritional factors and can improve the functioning of individual systems and the body as a whole. The main source of ω-3 PUFA has long been fish fat, which contains PUFA in the triglyceride form. A fairly new and promising alternative to fish fat is the liver fat of the Commander squid (Berryteuthis magister), which additionally contains alkylglycerols, contains PUFA in the phospholipid form and can be obtained from squid fishery waste. The objective of the research was to carry out an analysis of scientific data, including the results of studies of the biological activity of squid fat, as well as its components that are part of other similar raw materials, and evaluate the prospects for its use in medical practice. Material and methods. During the study, various sources were analyzed, including scientific literature from electronic databases eLibrary, PubMed, Scopus, Web of Science and electronic search engines Google Academy from 2000 to 2022. Results. It is noted that squid fat has a pronounced biological activity. Its components increase innate immunity, have antitumor potential, improve the state of the body under stress, have hypolipidemiс and hypotensive effect, improve memory and attention, and also positively affect the composition and rheological parameters of blood. In addition to these effects, a positive effect of ω-3 PUFA and alkylglycerols on spermatogenesis, sperm quality and the female reproductive system has been noted. In a number of studies, alkylglycerol esters increased the permeability of the blood-brain barrier and, due to their structure, are able to form vesicles, therefore, they can be considered as raw materials for the production of new dosage forms for targeted therapy of brain tumors. In the available literature, in the case of the use of squid fat and its components, undesirable side effects have not been identified. Conclusion. Squid fat is a complete source of ω-3 PUFA and alkylglycerols, therefore it can be recommended as a dietary supplement, especially in a diet low in ω-3 PUFA.

In silico evaluation of marine fish proteins as nutritional supplements for COVID-19 patients.

To date, no specific drug has been discovered for the treatment of COVID-19 and hence, people are in a state of anxiety. Thus, there is an urgent need to search for various possible strategies including nutritional supplementation. In this study, we have tried to provide a reference for protein supplementation. Specifically, 20 marine fish proteins were subjected to in silico hydrolysis by gastrointestinal enzymes, and a large number of active peptides were generated. Then, the binding abilities of these peptides to SARS-CoV-2 main protease and monoamine oxidase A were assessed. The results showed that NADH dehydrogenase could be a good protein source in generating potent binders to the two enzymes, followed by cytochrome b. In addition, some high-affinity oligopeptides (VIQY, ICIY, PISQF, VISAW, AIPAW, and PVSQF) were identified as dual binders to the two enzymes. In summary, the supplementation of some fish proteins can be helpful for COVID-19 patients; the identified oligopeptides can be used as the lead compounds to design potential inhibitors against COVID-19 and anxiety.

Discovery of Novel Angiotensin-Converting Enzyme Inhibitory Peptides from Todarodes pacificus and Their Inhibitory Mechanism: In Silico and In Vitro Studies.

In order to rapidly and efficiently excavate antihypertensive ingredients in Todarodes pacificus, its myosin heavy chain was hydrolyzed in silico and the angiotensin-converting enzyme (ACE) inhibitory peptides were predicted using integrated bioinformatics tools. The results showed the degree of hydrolysis (DH) theoretically achieved 56.8% when digested with papain, ficin, and prolyl endopeptidase (PREP), producing 126 ACE inhibitory peptides. By predicting the toxicity, allergenicity, gastrointestinal stability, and intestinal epithelial permeability, 30 peptides were finally screened, of which 21 had been reported and 9 were new. Moreover, the newly discovered peptides were synthesized to evaluate their in vitro ACE inhibition, showing Ile-Ile-Tyr and Asn-Pro-Pro-Lys had strong effects with a pIC50 of 4.58 and 4.41, respectively. Further, their interaction mechanisms and bonding configurations with ACE were explored by molecular simulation. The preferred conformation of Ile-Ile-Tyr and Asn-Pro-Pro-Lys located in ACE were successfully predicted using the appropriate docking parameters. The molecular dynamics (MD) result indicated that they bound tightly to the active site of ACE by means of coordination with Zn(II) and hydrogen bonding and hydrophobic interaction with the residues in the pockets of S1 and S2, resulting in stable complexes. In summary, this work proposed a strategy for screening and identifying antihypertensive peptides from Todarodes pacificus.

Mercury Concentrations and Se:Hg Molar Ratios in Flyingfish (Exocoetus volitans) and Squid (Uroteuthis chinensis).

We measured total mercury (Hg) and selenium (Se) concentrations as well as stable nitrogen (N) isotopic composition in flyingfish and squid muscle tissues from the eastern Indian Ocean and western South China Sea. The results showed that the mean Hg concentration in squid muscle from the western South China Sea was lower than that in the eastern Indian Ocean. The Hg concentrations in flyingfish and squid muscle samples were positively correlated with organism size (length and weight) and δ15N in all the study areas. Furthermore, we found a negative correlation between Se and Hg in molar content of flyingfish and squid muscle from the western South China Sea. The Se:Hg molar ratio was significantly negative correlated with fish weight and δ15N, suggesting that the Se:Hg molar ratio decreases with the increase of fish size and trophic level in the food web.

Effects of melanin from Sepiella Maindroni ink (MSMI) on the intestinal Microbiome of mice.

BACKGROUND: By the search for new natural compounds with beneficial health effects, cephalopod ink has been considered as an attempt to develop new drugs and functional foods, which is an especially active field in Asia, where cephalopods are a major fishery catch, for which ink sacs are a bi-product and where homeopathic medicine has deep roots. There is a demand to evaluate the safety and influence to the organism. The specific composition and relative abundance of the gut microbiota, which is potentially a major modulator of host metabolism, drives the interaction between functional foods and host health. We explore the effects of melanin from Sepiella Maindroni, most common cuttlefish in China, on the intestinal microbiome of mice. RESULTS: ICR mice were randomly divided four groups, which were normal group (S), low melanin dose group (D; 120 mg/kg), medium melanin dose group (Z; 240 mg/kg), and high melanin dose group (G; 480 mg/kg). Melanin was delivered for 28 consecutive days. Fecal samples were used to generate 7715 operational taxonomic units (OTUs) via high-throughput sequencing. There were significant shifts in relative abundance of the dominant taxa at the phylum, class, order, family, and genus levels following melanin treatment. CONCLUSIONS: MSMI had no significant effect on the structure of intestinal flora in mice. The main effect was in the proportion of dominant bacterial communities. The effect positively correlated with the dose. From a health point of view, the use of melanin does not cause intestinal flora disorder. Our results may have important implications for MSMI as functional food component and potential therapeutic for manipulating gut microbiota.

Glucomannan or Glucomannan Plus Spirulina-Enriched Squid-Surimi Diets Reduce Histological Damage to Liver and Heart in Zucker fa/fa Rats Fed a Cholesterol-Enriched and Non-Cholesterol-Enriched Atherogenic Diet.

Glucomannan-enriched squid surimi improves cholesterolemia and liver antioxidant status. The effect of squid surimi enriched with glucomannan or glucomannan plus spirulina on liver and heart structures and cell damage markers was tested in fa/fa rats fed highly saturated-hyper-energetic diets. Animals were fed 70% AIN-93M rodent diet plus six versions of 30% squid surimi for 7 weeks: control (C), glucomannan (G), and glucomannan plus spirulina (GS). The cholesterol-control (HC), cholesterol-glucomannan (HG), and cholesterol-glucomannan plus spirulina (HGS) groups were given similar diets that were enriched with 2% cholesterol and 0.4% cholic acid. G and GS diets versus C diet significantly inhibited weight gain and lowered plasma alanine aminotransferase and aspartate aminotransferase, liver steatosis, lipogranulomas, and total inflammation and alteration scores. The hypercholesterolemic agent significantly increased the harmful effects of the C diet. Liver weight, the hepatosomatic index, all damage markers, and total histological scoring rose for HC versus C (at least P < .05). The addition of glucomannan (HG vs. HC) improved these biomarkers, and non-additional effects from spirulina were observed except for the total liver alteration score. In conclusion, glucomannan and glucomannan plus spirulina blocked the highly saturated-hyper-energetic diet negative effects both with and without added cholesterol. Results suggest the usefulness of including these functional ingredients in fish products.

Levels of ²¹°Po and ²¹°Pb in fish and molluscs in Slovenia and the related dose assessment to the population.

²¹°Po and ²¹°Pb activity concentrations in fish from the Slovenian part of Adriatic Sea, in the vicinity of a former uranium mine at Zirovski vrh and from the Slovenian market were determined. In addition, ²¹°Po and ²¹°Pb activity concentrations in squid from the Slovenian market and in mussels from the Slovenian part of the Adriatic Sea were also determined. Fish, squid and mussel consumption in Slovenia was assessed from the data available from Eurostat and Food and Agriculture Organisation (FAO) and the data used for the corresponding dose calculation. Fish species with the highest activity concentrations were grilled to assess possible loss of ²¹°Po during the food preparation process. Samples were freeze dried and radiochemical separation of ²¹°Po and ²¹°Pb was performed. Measurements of ²¹°Po were performed by alpha spectrometry and ²¹°Pb by a low background gas-flow proportional counter. ²¹°Po activity concentrations in fish, squid and mussels were from 0.039 to 35.0 Bqkg⁻¹ fresh weight and ²¹°Pb activity concentrations were from 0.08 to 3.03 Bqkg⁻¹ fresh weight. Grilling of fish resulted in no significant loss of ²¹°Po at 90°C. The assessed combined annual effective ingestion dose due to ²¹°Po and ²¹°Pb for fish, squid and mussels consumed in Slovenia is 47.6 µSv year⁻¹.

Levels of total mercury in marine organisms from Adriatic Sea, Italy.

The presence of total mercury in fish, crustacean and cephalopod from Adriatic Sea, was investigated. The highest concentrations were observed in decreasing order in: Norway lobster (0.97 +/- 0.24 mg/kg; mean +/- SE), European hake (0.59 +/- 0.14 mg/kg), red mullet (0.48 +/- 0.09 mg/kg), blue whiting (0.38 +/- 0.09 mg/kg), Atlantic mackerel (0.36 +/- 0.08 mg/kg) and European flying squid (0.25 +/- 0.03 mg/kg). A significant difference (p < 0.01) was found between the levels of total mercury in Norway lobster and those detected in all other species. The 25% of all samples exceeded the maximum limit fixed by Commission Regulation (EC) No 1881/2006. The results show that fish and fishery products can exceed the maximum levels and stress the need of more information for consumers in particular for people that eat large amount of fish.

Squid p196, a new member of the myosin-V class of motor proteins, is associated with motile axoplasmic organelles.

Axoplasmic organelles obtained from the squid giant axon move on actin filaments at an average velocity of 1 microm/s [Nature 356 (1992) 722]. The unconventional myosins, in particular, the myosin-V class of motor proteins, represent the most likely candidates to have a role in this motility. Experiments were performed to determine whether a member of the myosin-V class of unconventional myosins is present in axoplasm and optic lobes. Western blots of axoplasm probed with an affinity purified antibody to chicken brain myosin-V (CBM-V) showed cross-reactivity with a protein of Mr 196 kD (p196) which was subsequently purified from squid optic lobes using a modification of a protocol for the purification of CBM-V [Methods Enzymol. 298 (1998) 3; Cell 75 (1993) 215]. Western blots of CBM-V probed with an alpha-p196 polyclonal IgG showed cross-reactivity with CBM-V. Purified p196 has been found to be a calmodulin (CaM) binding protein that possesses calcium-stimulated actin-activated ATPase activity. Equilibrium density fractionation of motile axoplasmic organelle preparations has revealed that p196 cosedimented with the peak organelle fraction into Percoll gradients in the presence of cytochalasin B and ATP. Based on this evidence, we conclude that the p196 present in axoplasm and purified from optic lobes is a squid homolog of CBM-V and functions as a motor for fast transport of membranous organelles on actin filaments in neurons.

Molecular cloning and expression of a full-length cDNA encoding acetylcholinesterase in optic lobes of the squid Loligo opalescens: a new member of the cholinesterase family resistant to diisopropyl fluorophosphate.

Acetylcholinesterase cDNA was cloned by screening a library from Loligo opalescens optic lobes; cDNA sequence analysis revealed an open reading frame coding for a protein of 610 amino acids that showed 20-41% amino acid identity with the acetylcholinesterases studied so far. The characteristic structure of cholinesterase (the choline binding site, the catalytic triad, and six cysteines that form three intrachain disulfide bonds) was conserved in the protein. The heterologous expression of acetylcholinesterase in COS cells gave a recovery of acetylcholinesterase activity 20-fold higher than in controls. The enzyme, partially purified by affinity chromatography, showed molecular and kinetic features indistinguishable from those of acetylcholinesterase expressed in vivo, which displays a high catalytic efficiency. Both enzymes are true acetylcholinesterase corresponding to phosphatidylinositol-anchored G2a dimers of class I, with a marked substrate specificity for acetylthiocholine. The deduced amino acid sequence may explain some particular kinetic characteristics of Loligo acetylcholinesterase, because the presence of a polar amino acid residue (S313) instead of a nonpolar one [F(288) in Torpedo] in the acyl pocket of the active site could justify the high substrate specificity of the enzyme, the absence of hydrolysis with butyrylthiocholine, and the poor inhibition by the organophosphate diisopropyl fluorophosphate.

A family of delayed rectifier Kv1 cDNAs showing cell type-specific expression in the squid stellate ganglion/giant fiber lobe complex.

Squid giant axons are formed by giant fiber lobe (GFL) neurons of the stellate ganglion (SG). Other large motoneurons in the SG form a parallel system. A small family of cDNAs (SqKv1A-D) encoding Kv1 alpha-subunits was identified in a squid (Loligo opalescens) SG/GFL library. Members have distinct 5' untranslated regions (UTRs) and initial coding regions, but beyond a certain point (nucleotide 34 of SqKv1A) only nine differences exist. 3' UTRs are identical. Predicted alpha-subunits are nearly identical, and only the N termini differ significantly, primarily in length. RNase protection assays that use RNA isolated from specific SG regions show that SqKv1A mRNA is expressed prominently in the GFL but not in the SG proper. SqKv1B yields the opposite pattern. SqKv1D also is expressed only in the SG. SqKv1C expression was not detectable. In situ hybridizations confirm these results and reveal that SqKv1B mRNA is abundant in many large neurons of the SG, whereas SqKv1D expression is limited to small isolated clusters of neurons. SqKv1A and B are thus the predominant Kv1 mRNAs in the SG/GFL complex. Activation properties of SqKv1A and B channels expressed in oocytes are very similar to one another and compare favorably with properties of native delayed rectifier channels in GFL neurons and large SG neurons. The Kv1 complement in these squid neurons thus seems to be relatively simple. Several differences exist between cloned and native channels, however, and may reflect differences in the cellular environments of oocytes and neurons.

A beta-subclass phosphatidylinositol-specific phospholipase C from squid (Loligo forbesi) photoreceptors exhibiting a truncated C-terminus.

A PCR-based strategy has been used to isolate a full length cDNA encoding a phosphatidylinositol-specific phospholipase C from a sized cDNA squid (Loligo forbesi) retinal library. The predicted protein sequence contains 875 amino acids, with calculated M(r) 98,181, and has marked similarity with PLC beta-isoforms, including conservation of the 'X' and 'Y' regions. It is unique in having a major C-terminal truncation. A major protein of apparent M(r) 120,000 estimated by SDS-PAGE has been isolated from squid photoreceptors and identified by partial protein sequence analysis to correspond to the protein sequence predicted from the cDNA clone. This protein has been shown to hydrolyse phosphatidylinositol 4,5-bisphosphate. It is not yet clear whether this represents the major light-activated PLC in squid vision.

Molecular characterization and functional expression of squid retinal-binding protein. A novel species of hydrophobic ligand-binding protein.

The primary structure of squid retinal-binding protein (RALBP) was determined by cDNA and protein sequencing. Squid RALBP contains 342 amino acid residues in a single N-terminal-blocked chain with a molecular weight of 39,111. The N alpha-blocking group was identified as an acetyl moiety by mass spectrometry. The amino acid sequence revealed that the protein is highly hydrophilic and acidic, but it has several hydrophobic regions that are located mainly in the middle part of the polypeptide chain. It is also predicted that these hydrophobic regions form beta-sheet structures. The primary structure of RALBP is, however, quite distinct from those of other retinoid-binding proteins, showing that squid RALBP is a novel hydrophobic ligand-binding protein that functions in intracellular retinoid transport. Using the cloned cDNA, squid RALBP was expressed in vitro. By carrying out the translation at 20 degrees C in reticulocyte lysates, the protein having retinol binding activity was produced.