IL-6, a Therapeutic Target and Omega-3 PUFA, a Host Modulator in Chronic Periodontitis

Periodontitis is a common multifactorial inflammatory disease with gradual loss of supportive tissues around the teeth which eventually leads to decrease in the quality of life. Blocking Interleukin-6 (IL-6), a multifunctional cytokine with pro-inflammatory properties has demonstrated therapeutic efficacy in inflammatory diseases like Rheumatoid arthritis, SLE and multiple sclerosis. Host immune response, the underlying cause for this progressive disease is targeted by Host modulatory therapy (HMT), an emerging treatment modality. Omega-3 polyunsaturated fatty acids (ώ 3 PUFAs), one of the relatively safe HMTs, reduces tissue destruction, stabilizes or even regenerates the periodontium through its anti-inflammatory & immunoregulatory properties. ώ 3 PUFAs are essential for the synthesis of eicosanoids which are involved in anti-inflammatory, antiplatelet aggregatory, vasodilation, vasoconstriction, immune response, cell growth and proliferation. The key factor examined and extrapolated in this study is the anti-inflammatory property of ώ 3 PUFA. The aim of the study was to evaluate the immunological and clinical response to ώ 3 PUFA supplementation therapy in chronic periodontitis by measuring the inflammatory cytokine, IL-6 levels in serum. In this open label exploratory study, 40 patients with a Female: Male ratio of 4:1were enrolled and assessed clinically by measuring Oral Hygiene Index-Simplified (OHI-S), Probing Pocket Depth (PPD), Clinical Attachment Level (CAL) and their serum for IL-6 levels. Subsequently 300 mg (concentration of EPA 180/DHA120) of ώ 3 PUFA was prescribed twice daily for 3 months and periodically reviewed to assess their IL-6 levels and periodontal status. IL-6 levels which were at a maximum mean of 10.2 pg/ml prior to treatment, showed a gradual and notable reduction to 2.3 pg/ml at the end of the study following ώ 3 PUFA supplementation therapy. The coefficient of variation R2 and ANOVA showed statistically significant periodic variation in biomarker IL-6 and in all clinical measurements at all time intervals. ώ 3 PUFA adjunctive therapy significantly reduces the inflammatory cytokine (IL-6) levels and causes noteworthy improvement of the most relevant clinical parameters (OHI-S, PPD, CAL). Hence ώ 3 PUFA can be recommended as a dietary supplementation and a safe host modulatory treatment in chronic periodontitis.

Dietary Omega-3 Polyunsaturated Fatty-Acid Supplementation Upregulates Protective Cellular Pathways in Patients with Type 2 Diabetes that reduces Painful Diabetic Neuropathy

Background: Omega-3 polyunsaturated fatty acids (PUFAs) are increasingly reported to improve chronic neuroinflammatory diseases in peripheral and central nervous systems. Specifically, docosahexaenoic acid (DHA) protects nerve cells from noxious stimuli in vitro and in vivo. Recent reports link PUFA supplementation to improving painful diabetic neuropathy (pDN) symptoms. However, the molecular mechanism behind omega-3 PUFAs ameliorating pDN symptoms is lacking. Therefore, we sought to determine the distinct cellular pathways that omega-3 PUFAs dietary supplementation promotes in reducing painful neuropathy in type 2 diabetes mellitus (DM2) patients. Methods: Forty volunteers diagnosed with type 2 diabetes were enrolled in the "En Balance-PLUS" diabetes education study. The volunteers participated in weekly lifestyle/nutrition education and daily supplementation with 1,000 mg DHA and 200 mg eicosapentaenoic acid. The Short-Form McGill Pain Questionnaire validated clinical determination of baseline and post-intervention pain complaints. Laboratory and untargeted metabolomics analyses were conducted using blood plasma collected at baseline and after three months of participation in the dietary regimen. The metabolomics data was analyzed using random forest, hierarchical cluster, ingenuity pathway analysis, and metabolic pathway mapping. Results: We found that metabolites involved in oxidative stress and glutathione production shifted significantly to a more anti-inflammatory state post supplementation. Example of these metabolites include cystathionine (+90%), S-methylmethionine (+9%), glycine cysteine-glutathione disulfide (+157%) cysteinylglycine (+19%), glutamate (-11%), glycine (+11%) and arginine (+13.4%). In addition, the levels of phospholipids associated with improved membrane fluidity such as linoleoyl-docosahexaenoyl-glycerol (18:2/22:6) (+253 %) were significantly increased. Ingenuity pathway analysis suggested several key bio functions associated with omega-3 PUFA supplementation such as formation of reactive oxygen species (p = 4.38 × 10-4, z-score = -1.96), peroxidation of lipids (p = 2.24 × 10-5, z-score = -1.944), Ca2+ transport (p = 1.55 × 10-4, z-score = -1.969), excitation of neurons (p = 1.07 ×10-4, z-score = -1.091), and concentration of glutathione (p = 3.06 × 10-4, z-score = 1.974). Conclusion: The reduction of pro-inflammatory and oxidative stress pathways following omega-3 PUFAS supplementation is consistent with using omega-3 PUFAs as a complementary dietary strategy as part of the overall treatment of painful diabetic neuropathy.

Anticancer Activity of Ω-6 Fatty Acids through Increased 4-HNE in Breast Cancer Cells

Her2-amplified breast cancers resistant to available Her2-targeted therapeutics continue to be a challenge in breast cancer therapy. Dox is the mainstay of chemotherapy of all types of breast cancer, but its usefulness is limited by cumulative cardiotoxicity. Because oxidative stress caused by dox generates the pro-apoptotic Ω-6 PUFA metabolite 4-hydroxynonenal (4-HNE), we surmised that Ω-6 PUFAs would increase the effectiveness of dox chemotherapy. Since the mercapturic acid pathway enzyme RALBP1 (also known as RLIP76 or Rlip) that limits cellular accumulation of 4-HNE also mediates dox resistance, the combination of Ω-6 PUFAs and Rlip depletion could synergistically improve the efficacy of dox. Thus, we studied the effects of the Ω-6 PUFA arachidonic acid (AA) and Rlip knockdown on the antineoplastic activity of dox towards Her2-amplified breast cancer cell lines SK-BR-3, which is sensitive to Her2 inhibitors, and AU565, which is resistant. AA increased lipid peroxidation, 4-HNE generation, apoptosis, cellular dox concentration and dox cytotoxicity in both cell lines while sparing cultured immortalized cardiomyocyte cells. The known functions of Rlip including clathrin-dependent endocytosis and dox efflux were inhibited by AA. Our results support a model in which 4-HNE generated by AA overwhelms the capacity of Rlip to defend against apoptosis caused by dox or 4-HNE. We propose that Ω-6 PUFA supplementation could improve the efficacy of dox or Rlip inhibitors for treating Her2-amplified breast cancer.

A novel treatment protocol in severe acute pancreatitis: the first double blinded comparative trial to assess efficiency of omega 3 fatty acid infusion

Background: For decades, the treatment of acute pancreatitis (AP) has been largely supportive. However, a therapeutic window for intervention using immune modulators exists between the onset of clinical symptoms and peak pro-inflammatory cytokine expression. Soybean oil derived ω-3 PUFA is shown to inhibit the formation of ω-6 PUFA derived pro-inflammatory eicosanoids and suppresses pro-inflammatory activity of nuclear transcription factors (NF-κβ); thus, bringing about a much-needed immune regulation, both locally and systemically. Our aim was to study the efficacy of ω-3 PUFA infusion in severe AP (SAP) and set a new treatment protocol.Methods: This, first of its kind, doubled blinded randomised control trial was undertaken to assess the efficacy of early ω-3 PUFA supplementation in patients with predicted SAP by using serum amylase, serum lipase and C-reactive protein (CRP) with BISAP and Marshal scoring systems as markers for clinical outcome. 60 such patients were randomised into two groups: 30 were given ω-3 PUFA infusion and 30 were given a placebo. Chi square test and unpaired t test were used for comparison.Results: ω-3 PUFA infusion was found to be highly significant (p<0.05) in bringing about clinical improvement, reduced progression of SAP, reduced hospital stay and prompt reversal of organ dysfunction and SIRS.Conclusions: ω-3 PUFA infusion is the future for the treatment of patients with SAP. This drug is cheap and easily available, has no known side effects, reduces the morbidity and mortality, reduces the duration of hospital stay; thus, resulting in overall reduced medical expenditure.

Polyunsaturated Fatty Acids, Exercise, and Cancer-Related Fatigue in Breast Cancer Survivors

Cancer-related fatigue (CRF) is one of the most frequently reported and disabling symptoms in cancer survivors. With its negative impact on the activities of daily living, work, social activities, and mood, CRF causes severe impairment of quality of life. A previous study showed that omega-6 polyunsaturated fatty acid (PUFA) supplementation unexpectedly reduced CRF compared with omega-3 PUFA supplementation and that omega-6 PUFA supplementation reduced pro-inflammatory serum markers in fatigued American breast cancer survivors. Meanwhile, a recent meta-analysis of individual patient data revealed significant benefits of exercise interventions on CRF. Recently, we completed our randomized controlled trial among early-stage Japanese breast cancer survivors, in which we examined the effect of baseline blood PUFA characteristics on change in CRF during the 12-week trial by exercise group and confirmed that increased Cancer Fatigue Scale (CFS) was associated with both docosahexaenoic acid (DHA) (p = 0.06) and omega-3 index (p = 0.08) at baseline in all participants (n = 46, omega-6/omega-3 ratio = 6.79, SD = 1.90). On the contrary, DHA at baseline was positively correlated with change in CRF (r = 0.40, p = 0.06) in the control group (n = 24, omega-6/omega-3 ratio = 7.0). Moreover, eicosapentaenoic acid (EPA) at baseline was positively correlated with leg strength (r = 0.39, p = 0.10) in the exercise group. In conclusion, blood PUFA balance might be associated with the effect of exercise on CRF. In addition, higher EPA in individuals who conducted exercise likely has a beneficial effect on muscle strength. Further investigation is needed to clarify the interaction between PUFAs and exercise for alleviating CRF.

Changes in EPA and DHA during supplementation with omega-3 fatty acids and incident cardiovascular events: secondary analysis from the OMEMI trial

Background In the OMEMI trial, elderly post-MI patients did not achieve reduction in cardiovascular events from supplementation of 1.8g n-3 polyunsaturated fatty acids (PUFA). In two recent trials of hypertriglyceridaemic patients the REDUCE-IT trial demonstrated an association between high levels of serum eicosapentaenoic acid (EPA) and reduced risk of CV events with 4 g/day icosapent ethyl supplements while in the STRENGTH trial no such association was present in patients treated with 4 g/day of EPA+ docosahexaenoic acid (DHA). Purpose To assess associations between changes in concentrations of EPA and DHA during two years supplementation with n-3 PUFA and incident cardiovascular events in the OMEMI trial. Methods In the randomized controlled OMEMI trial, 1014 elderly patients with a recent acute myocardial infarction (AMI) were treated with 1.8g/day of EPA and docosahexaenoic acid (DHA) or placebo for two years, and followed for the primary outcome of MACE (AMI, coronary revascularization, stroke or heart failure hospitalization) and secondary outcome of new-onset atrial fibrillation (AF). Serum concentrations of EPA and DHA were measured at inclusion and at study completion by gas chromatography, and reported as % weight of total FA (%wt) in serum phospholipids. Results Serial EPA and DHA measurements at study inclusion and completion were available in 881 patients (92% of survivors). At baseline EPA and DHA concentrations were (mean±SD) 2.84±1.41 and 5.71±1.35%wt, respectively. Higher baseline EPA and DHA concentrations were associated with previous n-3 PUFA supplementation, lower prevalence of current smoking and diabetes, lower levels of triglycerides and higher levels of HDL-cholesterol (all p&lt;0.05). In patients randomized to n-3 PUFA, EPA and DHA increased with 2.32±1.92 and 0.91±1.19%wt, respectively, whereas in the placebo group EPA and DHA decreased with −0.39±1.37 and −0.43±1.13%wt, respectively. Greater increases in EPA and DHA during follow-up were associated with a lowering of triglyceride concentrations, increasing HDL concentrations, and lower baseline concentrations of EPA and DHA (all p&lt;0.001). Among patients treated with n-3 PUFA (n=438), a greater increase in EPA was associated with a lower risk of incident MACE (HR 0.89 [95% CI 0.78–1.00] per %wt, p=0.059) and higher risk of new-onset AF in patients free of AF at inclusion (n=339): HR 1.31 [1.06–1.62] per %wt, p=0.012 (Figure). There were no such associations for changes in DHA: HR 0.86 (95% CI 0.70–1.05), p=0.13 for MACE and HR 1.29 (0.91–1.83), p=0.16 for new-onset AF. Conclusion Patients treated with 1.8 g/day n-3 PUFA for two years experienced a doubling of serum EPA concentrations. Greater increases in EPA were associated with a lower risk of MACE, but also a higher risk of new-onset AF. Changes in DHA concentrations were not associated with outcomes, suggesting that EPA may be the more important n-3 PUFA with respect to risk of cardiovascular events. FUNDunding Acknowledgement Type of funding sources: Public hospital(s). Main funding source(s): Oslo University Hospital, Ullevål Figure 1

Effects of Polyunsaturated Fatty Acids Supplementation on the Meat Quality of Pigs: A Meta-Analysis

Polyunsaturated fatty acids (PUFAs) supplementation has been widely discussed as a strategy for improving meat quality in pig production, but the effects are inconsistent. This meta-analysis was performed to comprehensively evaluate its effects on the meat quality and growth performance of pigs. We searched the PubMed and the Web of Science databases (articles published from January 1, 2000 to October 16, 2020) and compared PUFAs-supplemented diets with control diets. We identified 1,670 studies, of which 14 (with data for 752 pigs) were included in our meta-analysis. The subgroup analysis was classified as PUFA source [conjugated linoleic acid (CLA) or linseed], concentration (high or low concentration), and initial stage (growing or finishing pigs). Our analysis found that PUFA supplementation increased the intramuscular fat (IMF) content (WMD = 0.467%, 95% CI: 0.312–0.621, p &lt; 0.001), decreased the meat color L* (WMD = −0.636, 95% CI: −1.225 to −0.047, p = 0.034), and pH 24 h (WMD = −0.021, 95% CI: −0.032 to −0.009, p &lt; 0.001) but had no influence on drip loss, meat color a* and b*, pH 45 min, and growth performance. CLA supplementation improved IMF content (WMD = 0.542%, 95% CI: 0.343–0.741, p &lt; 0.001) and reduced meat color b* (WMD = −0.194, 95% CI: −0.344 to −0.044, p = 0.011). Linseed supplementation increased IMF content (WMD = 0.307%, 95% CI: 0.047–0.566, p = 0.021), decreased meat color L* (WMD = −1.740, 95% CI: −3.267 to −0.213, p = 0.026), and pH 24 h (WMD = 0.034, 95% CI: −0.049 to −0.018, p &lt; 0.001). We discovered an increase on the IMF content in both high and low concentration PUFA supplementation (WMD = 0.461%, 95% CI: −0.344 to −0.044, p &lt; 0.001; WMD = 0.456%, 95% CI: 0.276–0.635, p &lt; 0.001). Furthermore, we also found the effects of PUFA supplementation on meat color L* and pH 24 h are concentration- and stage-dependent. PUFA supplementation can improve the meat quality of pigs, which mainly emerges in greatly increasing IMF content.

Zinc Deficiency, Plasma Fatty Acid Profile and Desaturase Activities in Hemodialysis Patients: Is Supplementation Necessary?

Background: Desaturation and elongation are critical processes in endogenous metabolic fatty acid pathways. Zinc (Zn) is a cofactor for desaturases and elongases enzymes. There is limited evidence regarding the relationships between biomarkers of Zn status, nutritional intake, plasma phospholipid fatty acid profile and clinical outcomes among patients undergoing hemodialysis (HD).Objective: To examine the relationships between dietary and serum levels of Zn and Cu/Zn ratio and to explore associations of these micronutrients with PUFA profile and estimated desaturase and elongase enzyme activities in serum phospholipids among HD patients.Methods: This study included 40 adult patients undergoing hemodialysis treatment. Repeated 24-h recalls were applied for dietary intake assessment. Serum concentration of Zn and Cu were determined using inductively coupled plasma mass spectrometry and fatty acid composition by gas-liquid chromatography. Desaturase and elongase activities were calculated from product-precursor fatty acid ratios.Results: Inadequate dietary Zn intake was found in 55% of HD patients. They all had serum Zn concentration below the reference value of 60 μg/dL (mean 38.8 ± 7.72 μg/dL). Adequate zinc intake was accompanied with significantly higher intake of energy, total fats, SFA, MUFA and proteins. There was no correlation between Zn serum status and Zn intake estimates. Serum Cu/Zn ratio was high, (2.76 ± 0.68), directly and significantly associated with HD period, CRP, BMI, VFA, and inversely with Kt/V, albumin, iron, and iPTH. The n-6/n-3 ratio in plasma phospholipids was elevated (12.25 ± 3.45) and patients with inadequate Zn intake had lower n-3 PUFA intake and status compared to those with adequate intake. Serum Zn concentrations were inversely correlated with linoleic/dihomo-γ-linolenic acid ratio (LA/DGLA) (p = 0.037), related to D6-desaturase activity (p = 0.033) and directly with DGLA relative abundances (p = 0.024). Cu status was inversely associated with EPA level (p = 0.03) and estimates of elongase activity (p = 0.001). Furthermore, positive relationship was found between the Cu/Zn ratio and determined elongase value (p = 0.01).Conclusion: Findings of this study underpin the high prevalence of Zn deficiency and inadequate n-3 PUFA intake and status among subjects undergoing HD. The results obtained indicate that the assessment of Zn status should be a standard parameter of nutritional status screening in HD patients while emphasizing the importance of Cu/Zn determination. Although further research is warranted, Zn and-n-3 PUFA supplementation in HD patients might be beneficial for the prevention and attenuation of adverse health outcomes

APOE Genotype Modifies the Plasma Oxylipin Response to Omega-3 Polyunsaturated Fatty Acid Supplementation in Healthy Individuals

The omega-3 polyunsaturated fatty acids (n-3 PUFAs), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), mediate inflammation in large part by affecting pro-inflammatory and anti-inflammatory/pro-resolving oxylipin concentrations. Common gene variants are thought to underlie the large inter-individual variation in oxylipin levels in response to n-3 PUFA supplementation, which in turn is likely to contribute to the overall heterogeneity in response to n-3 PUFA intervention. Given its known role in inflammation and as a modulator of the physiological response to EPA and DHA, here we explore, for the first time, the differential response of plasma hydroxy-, epoxy- and dihydroxy-arachidonic acid, EPA and DHA oxylipins according to apolipoprotein E (APOE) genotype using samples from a dose-response parallel design RCT. Healthy participants were given doses of EPA+DHA equivalent to intakes of 1, 2, and 4 portions of oily fish per week for 12 months. There was no difference in the plasma levels of EPA, DHA or ARA between the wildtype APOE3/E3 and APOE4 carrier groups after 3 or 12 months of n-3 PUFA supplementation. At 12 months, hydroxy EPAs (HEPEs) and hydroxy-DHAs (HDHAs) were higher in APOE4 carriers, with the difference most evident at the highest EPA+DHA intake. A significant APOE*n-3 PUFA dose effect was observed for the CYP-ω hydroxylase products 19-HEPE (p = 0.027) and 20-HEPE (p = 0.011). 8-HEPE, which, along with several other plasma oxylipins, is an activator of peroxisome proliferator activated receptors (PPARs), showed the highest fold change in APOE4 carriers (14-fold) compared to APOE3/E3 (4-fold) (p = 0.014). Low basal plasma EPA levels (EPA &lt; 0.85% of total fatty acids) were associated with a greater change in 5-HEPE, 9-HEPE, 11-HEPE, and 20-HEPE compared to high basal EPA levels (EPA &gt; 1.22% of total fatty acids). In conclusion, APOE genotype modulated the plasma oxylipin response to increased EPA+DHA intake, with APOE4 carriers presenting with the greatest increases following high dose n-3 PUFA supplementation for 12 months.

Fish Oil for Healthy Aging: Potential Application to Master Athletes

Master athletes perform high volumes of exercise training yet display lower levels of physical functioning and exercise performance when compared with younger athletes. Several reports in the clinical literature show that long chain n-3 polyunsaturated fatty acid (LC n-3 PUFA) ingestion promotes skeletal muscle anabolism and strength in untrained older persons. There is also evidence that LC n-3 PUFA ingestion improves indices of muscle recovery following damaging exercise in younger persons. These findings suggest that LC n-3 PUFA intake could have an ergogenic effect in master athletes. However, the beneficial effect of LC n-3 PUFA intake on skeletal muscle in response to exercise training in both older and younger persons is inconsistent and, in some cases, generated from low-quality studies or those with a high risk of bias. Other factors such as the choice of placebo and health status of participants also confound interpretation of existing reports. As such, when considered on balance, the available evidence does not indicate that ingestion of LC n-3 PUFAs above current population recommendations (250–500 mg/day; 2 portions of oily fish per week) enhances exercise performance or recovery from exercise training in master athletes. Further work is now needed related to how the dose, duration, and co-ingestion of LC n-3 PUFAs with other nutrients such as amino acids impact the adaptive response to exercise training. This work should also consider how LC n-3 PUFA supplementation may differentially alter the lipid profile of cellular membranes of key regulatory sites such as the sarcolemma, mitochondria, and sarcoplasmic reticulum.