Ida-Johanne Jensen

ω-3 Fatty Acids and Cardiovascular Diseases: Effects, Mechanisms and Dietary Relevance.

ω-3 fatty acids (n-3 FA) have, since the 1970s, been associated with beneficial health effects. They are, however, prone to lipid peroxidation due to their many double bonds. Lipid peroxidation is a process that may lead to increased oxidative stress, a condition associated with adverse health effects. Recently, conflicting evidence regarding the health benefits of intake of n-3 from seafood or n-3 supplements has emerged. The aim of this review was thus to examine recent literature regarding health aspects of n-3 FA intake from fish or n-3 supplements, and to discuss possible reasons for the conflicting findings. There is a broad consensus that fish and seafood are the optimal sources of n-3 FA and consumption of approximately 2–3 servings per week is recommended. The scientific evidence of benefits from n-3 supplementation has diminished over time, probably due to a general increase in seafood consumption and better pharmacological intervention and acute treatment of patients with cardiovascular diseases (CVD).

Changes in antioxidative capacity of saithe (Pollachius virens) and shrimp (Pandalus borealis) during in vitro digestion.

The health effects of seafood have primarily been linked to eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3). These omega-3 fatty acids are highly susceptible to oxidation. Peptides exhibiting antioxidative capacity (AOC) are encrypted in seafood muscle proteins. Such components may protect the fatty acids during digestion and uptake, as well as exhibit antioxidative effects in general. The oxygen radical absorbance capacity (ORAC) assay and the ferric reducing antioxidant power (FRAP) assay were used to study the changes in AOC of fish and shrimp muscle and their aqueous fractions, referred to as press juice (PJ), during a simulated in vitro gastrointestinal (GI) digestion. Blueberry (Vaccinium myrtillus), well-known for its AOC, was included for comparison. During digestion the AOC increased in all samples. After digestion the AOC of muscle of both autumn saithe and shrimp were higher (130-165 micromol/g), and winter saithe comparable (110 micromol/g) to digested blueberry when measured by ORAC. The AOC of PJ was low in general (5-20 micromol/g). When measured by FRAP, blueberry exhibited ten times the AOC of seafood muscle. Antioxidative compounds in seafood increased significantly during digestion, and may offer protection of other beneficial food components such as EPA and DHA.

Proximate composition, antihypertensive and antioxidative properties of the semimembranosus muscle from pork and beef after cooking and in vitro digestion

The aims of this study were to evaluate and compare proximate composition, antihypertensive activity and antioxidative capacity of the semimembranosus muscle from pork and beef and to study how these characteristics were affected by household preparation and subsequent digestion. The proximate composition was similar between pork and beef. Both pork and beef contained protein with the essential amino acids. Cooking in a heated pan did not affect the retention of lipid or sum of amino acids, but reduced the amount of the free amino acid taurine. The antihypertensive effect did not differ significantly between pork and beef, whereas the antioxidative capacity did. Cooking affected the antioxidative capacity negatively. The results from this study show that pork and beef are equally good sources of protein and bioactive properties, and whereas the nutritional composition is not affected, bioactive properties may be reduced after household preparations.

Preclinical and Clinical Studies on Antioxidative, Antihypertensive and Cardioprotective Effect of Marine Proteins and Peptides—A Review

High seafood consumption has traditionally been linked to a reduced risk of cardiovascular diseases, mainly due to the lipid lowering effects of the long chained omega 3 fatty acids. However, fish and seafood are also excellent sources of good quality proteins and emerging documentation show that, upon digestion, these proteins are sources for bioactive peptides with documented favorable physiological effects such as antioxidative, antihypertensive and other cardioprotective effects. This documentation is mainly from in vitro studies, but also animal studies are arising. Evidence from human studies evaluating the positive health effects of marine proteins and peptides are scarce. In one study, a reduction in oxidative stress after intake of cod has been documented and a few human clinical trials have been performed evaluating the effect on blood pressure. The results are, however, inconclusive. The majority of the human clinical trials performed to investigate positive health effects of marine protein and lean fish intake, has focused on blood lipids. While some studies have documented a reduction in triglycerides after intake of lean fish, others have documented no effects.

Dietary intake of cod and scallop reduces atherosclerotic burden in female apolipoprotein E-deficient mice fed a Western-type high fat diet for 13 weeks.

BackgroundIt is now increasingly recognized that the beneficial effects of seafood consumption is not limited to lipids and fatty acid, but that the protein part, i.e., peptides and amino acids, together with vitamins and even unknown bioactive constituents also are important for disease prevention. This study was designed to evaluate the putative anti-atherogenic effects of different protein sources (a lean seafood and a nonseafood) in apolipoprotein E-deficient (apoE−/−) mice.MethodsTwenty-four 5-week-old female apoE−/− mice were fed Western type diets containing chicken or a combination of cod and scallops as dietary protein sources for 13 weeks. Atherosclerotic plaque burden, weight, serum levels of leptin, glucose and LDL cholesterol as well as gene expressions from liver and heart were evaluated. Statistical analyses were performed using SPSS. Differences between the variables were evaluated using independent t-test or Mann–Whitney U test for normally and non-normally distributed variables, respectively. Normality was defined by the Shapiro-Wilk test.ResultsThe mice fed cod-scallop had a 24 % (p < 0.05) reduced total aorta atherosclerotic plaque burden compared to the chicken fed group, whereas the reduction in plaque in the less lesion prone thoracic and abdominal parts of the descending aorta were 46 % (p < 0.05) and 56 % (p < 0.05), respectively. In addition, mice fed cod-scallop gained less weight, and had lower serum levels of leptin, glucose and LDL cholesterol, compared to those fed chicken. Analysis of expression of the genes from liver and heart showed that hepatic endogenous antioxidant paraoxonase 2 (Pon2 gene) and the vascular cell adhesion molecule VCAM-1 (Vcam1 gene) were down regulated in mice fed cod-scallop compared to mice fed chicken.ConclusionThe present study revealed a metabolic beneficial effect of lean seafood compared to chicken, as atherosclerotic plaque burden, serum glucose, leptin and LDL cholesterol levels were reduced in mice fed cod-scallop.

The potential of cod hydrolyzate to inhibit blood pressure in spontaneously hypertensive rats

Hypertension is an independent yet controllable risk factor for cardiovascular diseases. Synthetic angiotensin-converting enzyme (ACE) inhibitors used to treat hypertension are often associated with adverse effects, and the interest in diet-related inhibitors is increasing. We hypothesized that North Atlantic fish hydrolysate might inhibit ACE, thus preventing hypertension. We assessed the ACE inhibitory potential of various North Atlantic fish species and evaluated the effect of dietary supplementation of fish hydrolysates on the blood pressure of spontaneously hypertensive rats. Fish samples were hydrolyzed using simulated gastrointestinal digestion, and ACE inhibitory activity was evaluated using an ACE inhibitory activity assay. In vivo anti-hypertensive effects were evaluated by administering hydrolysates of wild Atlantic cod (Gadus morhua L.), haddock (Melanogrammus aeglefinus L.), or farmed Atlantic salmon (Salmo salar L.) to 10-week-old male, spontaneously hypertensive rats for 4 weeks. The dosing was 200 mg/kg body weight for 21 days, followed by 500 mg/kg body weight for 7 days. Water and Captopril (20 mg/kg body weight) were administered as the negative and positive controls, respectively. The analyzed fish hydrolysates exhibited a 50% ACE inhibition coefficient (IC50) of 1 to 2.7 μg/mU ACE. Fish hydrolysate supplements did not significantly inhibit the increase in blood pressure during the experimental period. The group receiving cod supplement had a lower (not significant) increase in blood pressure compared to the other groups. Although further studies are necessary to verify the antihypertensive effect of cod, the results obtained in this study indicate the potential that cod hydrolysate may have in inhibiting hypertension.

Cellular and Chemical Assays for Discovery of Novel Antioxidants in Marine Organisms

Low-molecular weight antioxidants receive increased interest both as protectors from disease and therapeutics, but are also used in other applications such as sun screens and food additives. Antioxidants are present in all organisms and are well characterized in terrestrial organisms whereas little knowledge exists for marine organisms. Only in Norwegian Arctic and sub-Arctic waters, thousands of species (invertebrates, algae, bacteria) are yet to be chemically characterized. Here we describe cell-based antioxidant assays (CAA and CLPAA in microplate format, and the Comet assay) measuring both hydrophilic and lipophilic antioxidant activities of single compounds and extract mixtures. Contrary to our chemical methods (ORAC and FRAP), cell-based assays assess cell membrane permeability and operate under more physiologically relevant conditions. A brief overview of our group’s work on finding novel antioxidants in marine organisms including testing activities of semipurified natural product extracts, identification of active compounds, purification of compounds in greater amounts, as well as how antioxidants can be tested in animals and humans is also described.

Protein Determination—Method Matters

The reported protein content of foods depends on the analytical method used for determination, making a direct comparison between studies difficult. The aim of this study was to examine and compare protein analytical methods. Some of these methods require extraction preceding analysis. The efficacy of protein extraction differs depending on food matrices and thus extraction yield was determined. Overall, most analytical methods overestimated the protein contents. The inaccuracies were linked to indirect measurements, i.e., nitrogen determination and subsequent conversion to protein, or interference from other chemical substances. Amino acid analysis is the only protein analysis method where interfering substances do not affect the results. Although there is potential for improvement in regards to the hydrolysis method, we recommend that this method should be the preferred for food protein determination.

Enzymatic Pre-Treatment Increases the Protein Bioaccessibility and Extractability in Dulse (Palmaria palmata)

Several common protein extraction protocols have been applied on seaweeds, but extraction yields have been limited. The aims of this study were to further develop and optimize existing extraction protocols and to examine the effect of enzymatic pre-treatment on bioaccessibility and extractability of seaweed proteins. Enzymatic pre-treatment of seaweed samples resulted in a three-fold increase in amino acids available for extraction. Combining enzymatic pre-treatment with alkaline extraction resulted in a 1.6-fold increase in the protein extraction yield compared to a standard alkaline extraction protocol. A simulated in vitro gastrointestinal digestion model showed that enzymatic pre-treatment of seaweed increased the amount of amino acids available for intestinal absorption 3.2-fold. In conclusion, enzymatic pre-treatment of seaweeds is effective for increasing the amount of amino acids available for utilization and may thus be an effective means for increasing the utilization potential of seaweed proteins. However, both the enzymatic pre-treatment protocol and the protein extraction protocol need further optimization in order to obtain optimal cost-benefit and results from the in vitro gastrointestinal digestion model need to be confirmed in clinical models.

Antioxidant and Anti-Inflammatory Activities in Extracts from Minke Whale (Balaenoptera acutorostrata) Blubber

Intake of long-chain omega-3 polyunsaturated fatty acids (LC-n3-PUFA) is commonly recognized to reduce cardiovascular disease (CVD). In previous studies, cold-pressed whale oil (CWO) and cod liver oil (CLO) were given as a dietary supplement to healthy volunteers. Even though CWO contains less than half the amount of LC-n3-PUFA of CLO, CWO supplement resulted in beneficial effects on anti-inflammatory and CVD risk markers compared to CLO. In the present study, we prepared virtually lipid-free extracts from CWO and CLO and evaluated the antioxidative capacity (AOC) and anti-inflammatory effects. Oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) assays were used to test the AOC, and the results indicated high levels of antioxidants present in all extracts. The anti-inflammatory effects of the extracts were tested with lipopolysaccharide- (LPS-) treated THP-1 cells, measuring its ability to reduce cytokine and chemokine secretion. Several CWO extracts displayed anti-inflammatory activity, and a butyl alcohol extract of CWO most effectively reduced TNF-α (50%, p < 0.05) and MCP-1 (85%, p < 0.001) secretion. This extract maintained a stable effect of reducing MCP-1 secretion (60%, p < 0.05) even after long-term storage. In conclusion, CWO has antioxidant and anti-inflammatory activities that may act in addition to its well-known LC-n3-PUFA effects.