We utilized high-performance liquid chromatography-mass spectrometry (HPLC-MS/MS), on dry achenes from four cultivars of Cannabis Sativa L.; CA, Cannabis Sativa L. Cultivar Amnesia; CB, Cannabis Sativa L. Cultivar Beldia; CM, Cannabis Sativa L. Cultivar Mexicana; CK, Cannabis Sativa L. Cultivar Khardala, to detect and quantify N-acylethanolamines (NAEs), which are bioactive compounds involved in lipid and energy metabolism. These plants were grown in Chefchaouen, northern Morocco. All four varieties displayed identical NAE lipid profiles, dominated by those derived from 16C and 18C fatty acids. In general, the NAE species presented the following concentration order: [LEA] > [OEA > POEA] > [SEA] > [PEA]. NAE-MUFA was the most abundant type, followed by NAE-PUFA and NAE-SFA, comprising 44, 37, and 19% of all NAEs, respectively, across the varieties. This research provides first-time quantification of NAEs in Cannabis achenes, thus enriching our understanding of these plants’ pharmaceutical and nutritional potential.

Objectives:Dietary n-3 polyunsaturated fatty acids (n-3PUFA)may influence the brain fatty acid (FA) profile and, thereby, the biosynthesis of FA bioactive metabolite, such as N-acylethanolamines (NAE). We investigated the influence of different dietary formulation of n-3PUFA from vegetable or marine oils on rat FA incorporation and modulation of NAE biosynthesis. Methods: Rats were fed for 10 weeks with diets containing 12% of fat from milk + 4% soybean oil and 4% of oils with different n-3 PUFA species: soybean oil as control (MilkFat), linseed oil with α-linolenic (ALA) (LSO), Buglossoides arvensis oil with ALA and stearidonic (SDA) (Buglos), fish oil with EPA and DHA (FO), Nannochloropsis microalga oil with EPA (Nanno) or Schizochytriummicroalga oil withDHA(Schy). Brain FA and NAE profile were assessed LC-DAD-MS/MS. Statistical Kruskal-Wallis test followed by Dunn’s correction was applied. Results: Different dietary n-3PUFA formulations induced FA change in the brain, compared to MilkFat group, DHA levels remained unchanged despite its elevated intake with FO and Schy diets. However, we found 9 fold (P < 0.001) increase of EPA with FO, arachidonic (AA) decreased significantly about 16% (P < 0.05) with LSO and FO diets. n-6 PUFA docosatetraenoic (DTA) (26% P < 0.05) and docosapentaenoic (DPA) (73% P < 0.05) were reduced with FO, probably due to a competition with EPA in the incorporation into phospholipids. We observed a 44% (P < 0.05) significant decrease of N-arachidonoylethanolamine (AEA)with LSO and FO, and a reduction of N-linoleoylethanolamine (LEA) by 49% (P< 0.05) with LSO. On the other hand, we found an increase of N-eicosapentanoylethanolamine (EPEA), derived from EPA, by 225% (P < 0.05) with FO, compared to MilkFat group. Conclusions: Our data indicate that reduction of AEA levels might attenuate its effects on the CB1 cannabinoid receptors, the main molecular target of the endogenous partial agonist AEA, which regulate physiological processes in the central nervous system. Therefore, in those pathophysiological conditions where is desirable to downregulate an overactive endocannabinoid system, a mixture of EPA and DHA as in FO, and ALA as in LSO, may potentially be a preferred nutritional source of n-3 PUFA. Funding Sources: University of Cagliari (FIR 2019) and Fundação FCT (Portugal).

Sustainable nutrition, equaling earth health, involves a personalized approach designed in terms of precision and avoidance of still cogent but unjustified dogmas, equaling public health. For instance, current dietary recommendations continue to dwell on the need to limit as much as possible the intake of saturated fatty acids (SFA), notwithstanding the mounting evidence that the effects of food on health cannot be predicted from the content of single nutrients without considering the overall macronutrient composition and the role of the food matrix. The traditional recommendation to restrict SFA ignores that their effects on health depend on the interaction between naturally occurring food components and those introduced by food processing. It is warranted to modify the still widely promoted dietary guidelines based upon such single nutrients as SFA and instead personalize dietary habits on the basis of the whole pattern of the food matrix. Accordingly, the double edge of malnutrition, that involves deficiency as well as excess and materializes in many individuals throughout their life course, might be tackled by implementing sustainability, with the additional effect of overcoming global inequalities. Within this context SFA may regain their position of tasty and cheap sources of energy to be adapted to each individual lifestyle.

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Numerous studies demonstrated the possibility to improve the properties of dairy products by increasing Polyunsaturated Fatty and lowering the concentrations of saturated ones. The present study aimed to examine the impact of the consumption of a dairy product (Pecorino cheese) rich in Conjugated Linoleic Acid (CLA) on lipid and health parameters in elderly subjects. During the first phase of the study, a Pecorino cheese rich in CLA was produced from milk of ewes fed a specific diet. In the second phase, a double blind randomized crossover intervention study was conducted to verify the impact of enriched Pecorino cheese on LDL-Cholesterol (LDL-Ch) in elder individuals. 58 subjects were enrolled in a nursing home in Padua (Italy) a, with a mean age of 85.9±6.0 years, 42 subjects (72.4%) were women. Study participants were randomly assigned to two groups: in one, they had a daily intake of up to 90g of Pecorino cheese rich in CLA (higher than 2.5%), in the other, they were given the same quantity of a Pecorino control cheese (CLA lower than 1%) for a two-month period. The active phase was followed by a one-month wash out period. The crossover was then carried out and participants ate the other type of cheese for two months. No significant differences were found in relation to LDL-Ch (estimated mean difference between group: 3.07 mg/dL, p=0.3171), while participants who consumed enriched cheese had a lower increase in glycaemia with respect to those assuming the control cheese (estimated mean difference between group: -0.07 g/L, p=0.0442). A significant carry-over effect was detected for HDL-Ch, tryglicerides, creatinine, sodium and vitamin D. Participants who consumed Pecorino cheese rich in CLA did not register an increase in lipid levels. More studies on humans are necessary to evaluate the role of CLA enriched cheese on cardiovascular protection.

The genetic polymorphisms in ewe and goat milk protein are of great importance as they are associated to quantitative and qualitative parameters and also may be responsible for the generation of bioactive peptides. In goat milk genetic polymorphisms also play a role in eliciting different degrees of allergic reaction in patients affected by cow milk protein allergy. Feeding strategies in lactating goats have been directed to investigate the relationship between diet and genotype. In sheep breeding feeding techniques as pasture, probiotic, vegetable and marine oils supplementation enhance nutritional properties of fat in milk, cheese and meat products. Technological strategies for cheese production have been set up to improve the health benefits trough probiotic and prebiotic adjunct. Experimental model and clinical studies have been performed to evaluate the impact of dairy products on human health.

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To extend our previous observations on the beneficial effect of dietary Pistacia lentiscus L. essential oil during cerebral bilateral common carotid artery occlusioninduced injury, we evaluated the activity of one of its major components, beta-caryophyllene (BCP), already known to possess peculiar biological activities, in Wistar rat cerebral cortex. Cerebral hypoperfusion was produced by a 30 min bilateral common carotid artery occlusion followed by 60 min reperfusion (BCCAO/R). Animals were starved for 12 hours before surgery and, 6 hours prior to hypoperfusion, BCP (40 mg/kg/0, 45 ml of sunflower oil as vehicle) was administered via gavage. Biological samples of brain tissue, plasma and cerebrospinal fluid (CSF) were examined by HPLC, western blot, gel zymography and immunohistochemistry and analyzed for fatty acids, expression of the enzyme ciclooxygenase-2 (COX-2), CB receptors for endocannabinoids (eCBs), and peroxisome proliferator-activated receptor (PPAR)-alpha and enzymatic activity of matrix-metalloprotease-9 (MMP9). Data obtained indicate that BCP appears to influence the outcome of BCCAO/R cerebral injury by modulating changes in levels of polyunsaturated fatty acids, biosynthesis of eCBs and eCB congeners, expression of CB1 and CB2 receptors, COX-2 protein levels and enzymatic activity of MMP9. Brain tissue response to the hypoperfusion/reperfusion-induced cerebral insult is modulated by dietary administration of BCP, suggesting the possible use of this molecule as nutritional treatment in neuroprevention. Work funded by Fondazione Banco di Sardegna.

Dietary fatty acids regulate several physiological functions. However, to exert their properties, they have to be present in the diet in an optimal balance. Particular attention has been focused on tissue highly polyunsaturated fatty acids (HPUFAs) n-6/n-3 ratio, influenced by the type and the esterified form of dietary fatty acids. Dietary EPA and DHA when esterified to phospholipids (PLs) are more efficiently incorporated into tissue PLs and seem to possess peculiar properties through specific mechanism(s) of action, such as the capacity to affect endocannabinoid biosynthesis at much lower doses than EPA and DHA in triglyceride form, probably because of the above mentioned higher incorporation into tissue PLs. Downregulation of the endocannabinoid system seems to mediate the positive effects exerted by omega-3-enriched PLs on several parameters of metabolic syndrome. PLs are one of the major dietary forms of EPA and DHA we are exposed to with the everyday diet; therefore, it is not surprising that it guarantees an effective EPA and DHA nutritional activity. Future studies should address whether EPA and DHA in PL form are also more effective than other formulations in ameliorating other pathological conditions where n-3 HPUFAs seem to exert beneficial activities such as cancer and psychiatric disorders.

In this study Pistacia lentiscus L. essential oil (E.O.), a mixture of terpenes and sesquiterpenes, was tested for its protective effects in cerebral ischemia/reperfusion-induced injury in Wistar rat frontal cortex and plasma. Cerebral ischemia was produced by a 20 min bilateral common carotid artery occlusion followed by 30 min reperfusion. Pistacia lentiscus L. essential oil (E.O.) (200 mg/0, 45 ml of sunflower oil as vehicle) was administered via gavage 6 hours prior to ischemia. Rats were randomly assigned to four groups, ischemic/reperfused (I/R) and sham-operated rats treated with the vehicle or with E.O.. Different brain areas were analysed for fatty acid changes and expression of the enzyme cyclooxygenase-2 (COX-2). Ischemia/reperfusion triggered in frontal cortex a decrease of docosahexaenoic acid (DHA), the membrane highly polyunsaturated fatty acid (HPUFA) most susceptible to oxidation. Pre-treatment with E.O. prevented this change and led further to decreased levels of COX-2, as assessed by Western Blot. In plasma of ischemic/reperfused rats, E.O. administration increased both the DHA-to-eicosapentaenoic acid (EPA) ratio and levels of the endocannabinoid congeners palmytoylethanolamide (PEA) and oleoylethanolamide (OEA). The results obtained suggest that ischemia/reperfusion triggers a cerebral insult sufficient to cause a a region specific lipid peroxidation as evidenced by the detectable, significant decrease in the tissue level of DHA, the most abundant essential fatty acid of neuronal membrane phospholipids. Acute dietary pre-treatment with E.O. triggers modifications both in the frontal cortex, where COX-2 expression decreases and the decrease of DHA is apparently prevented, and in plasma, where PEA and OEA levels increase. We suggest that the activity of PEA and OEA, as endogenous ligands of the peroxisome proliferator-activated receptor (PPAR)-alpha, by inducing the peroxisomal beta oxidation, may explain the observed increase in the DHA/EPA ratio. The latter, in fact, might account for an increased metabolism of n-3 aimed at restoring DHA within damaged brain tissue. The possibility that changes in fatty acid metabolism and plasmatic availability of PEA and OEA are correlated events represents an issue worth future investigations.