Ángel Luis López de Pablo

Cocoa has cardiovascular beneficial effects related to its content of antioxidant phytochemicals. Cocoa manufacturing produces large amounts of waste, but some by-products may be used as ingredients with health-promoting potential. We aimed to investigate the vasoactive actions of an extract from cocoa shell (CSE), a by-product containing theobromine (TH), caffeine (CAF) and protocatechuic acid (PCA) as major phytochemicals. In carotid and iliac arteries from 5-month and 15-month-old rats, we investigated CSE vasoactive properties, mechanism of action, and the capacity of CSE, TH, CAF and PCA to improve age-induced endothelial dysfunction. Vascular function was evaluated using isometric tension recording and superoxide anion production by dihydroethidium (DHE) staining and confocal microscopy. CSE caused endothelium-dependent vasorelaxation, blocked by L-NAME, but not indomethacin, regardless of sex, age, or vessel type. CSE maximal responses and EC₅₀ were significantly lower compared to acetylcholine (ACh). Arterial preincubation with CSE, TH, CAF or PCA, significantly reduced the number of vascular DHE-positive cells. Compared to adult males, iliac arteries from aged males exhibited reduced ACh concentration-dependent vasodilatation but larger CSE responses. In iliac arteries from aged male and female rats, preincubation with 10⁻⁴ M CSE and PCA, but not TH or CAF, improved ACh-relaxations. In conclusion, CSE has vasodilatory properties associated with increased nitric oxide bioavailability, related to its antioxidant phytochemicals, being particularly relevant PCA. Therefore, CSE is a potential food ingredient for diseases related to endothelial dysfunction.

Fetal undernutrition is a risk factor for cardiovascular diseases. Male offspring from rats exposed to undernutrition during gestation (MUN) exhibit oxidative stress during perinatal life and develop cardiac dysfunction in ageing. Angiotensin-II is implicated in oxidative stress-mediated cardiovascular fibrosis and remodeling, and lactation is a key developmental window. We aimed to assess if alterations in RAS during lactation participate in cardiac dysfunction associated with fetal undernutrition. Control dams received food ad libitum, and MUN had 50% nutrient restriction during the second half of gestation. Both dams were fed ad libitum during lactation, and male offspring were studied at weaning. We assessed: ventricular structure and function (echocardiography); blood pressure (intra-arterially, anesthetized rats); collagen content and intramyocardial artery structure (Sirius red, Masson Trichromic); myocardial and intramyocardial artery RAS receptors (immunohistochemistry); plasma angiotensin-II (ELISA) and TGF-β1 protein expression (Western Blot). Compared to Control, MUN offspring exhibited significantly higher plasma Angiotensin-II and a larger left ventricular mass, as well as larger intramyocardial artery media/lumen, interstitial collagen and perivascular collagen. In MUN hearts, TGF-β1 tended to be higher, and the end-diastolic diameter and E/A ratio were significantly lower with no differences in ejection fraction or blood pressure. In the myocardium, no differences between groups were detected in AT1, AT2 or Mas receptors, with MrgD being significantly lower in the MUN group. In intramyocardial arteries from MUN rats, AT1 and Mas receptors were significantly elevated, while AT2 and MrgD were lower compared to Control. Conclusions. In rats exposed to fetal undernutrition, RAS disbalance and associated cardiac remodeling during lactation may set the basis for later heart dysfunction.

Fetal undernutrition programs cardiometabolic diseases, with higher susceptibility in males. The mechanisms implicated are not fully understood and may be related to sex differences in placental adaptation. To evaluate this hypothesis, we investigated placental oxidative balance, vascularization, glucocorticoid barrier, and fetal growth in rats exposed to 50% global nutrient restriction from gestation day 11 (MUN, n = 8) and controls (n = 8). At gestation day 20 (G20), we analyzed maternal, placental, and fetal weights; oxidative damage, antioxidants, corticosterone, and PlGF (placental growth factor, spectrophotometry); and VEGF (vascular endothelial growth factor), 11β-HSD2, p22^phox, XO, SOD1, SOD2, SOD3, catalase, and UCP2 expression (Western blot). Compared with controls, MUN dams exhibited lower weight and plasma proteins and higher corticosterone and catalase without oxidative damage. Control male fetuses were larger than female fetuses. MUN males had higher plasma corticosterone and were smaller than control males, but had similar weight than MUN females. MUN male placenta showed higher XO and lower 11β-HSD2, VEGF, SOD2, catalase, UCP2, and feto-placental ratio than controls. MUN females had similar feto-placental ratio and plasma corticosterone than controls. Female placenta expressed lower XO, 11β-HSD2, and SOD3; similar VEGF, SOD1, SOD2, and UCP2; and higher catalase than controls, being 11β-HSD2 and VEGF higher compared to MUN males. Male placenta has worse adaptation to undernutrition with lower efficiency, associated with oxidative disbalance and reduced vascularization and glucocorticoid barrier. Glucocorticoids and low nutrients may both contribute to programming in MUN males.

One of the consequences of high altitude (hypobaric hypoxia) exposure is the development of right ventricular hypertrophy (RVH). One particular type of exposure is long-term chronic intermittent hypobaric hypoxia (CIH); the molecular alterations in RVH in this particular condition are less known. Studies show an important role of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex-induced oxidative stress and protein kinase activation in different models of cardiac hypertrophy. The aim was to determine the oxidative level, NADPH oxidase expression and MAPK activation in rats with RVH induced by CIH. Male Wistar rats were randomly subjected to CIH (2 days hypoxia/2 days normoxia; n = 10) and normoxia (NX; n = 10) for 30 days. Hypoxia was simulated with a hypobaric chamber. Measurements in the RV included the following: hypertrophy, Nox2, Nox4, p22phox, LOX-1 and HIF-1α expression, lipid peroxidation and H₂O₂ concentration, and p38α and Akt activation. All CIH rats developed RVH and showed an upregulation of LOX-1, Nox2 and p22phox and an increase in lipid peroxidation, HIF-1α stabilization and p38α activation. Rats with long-term CIH-induced RVH clearly showed Nox2, p22phox and LOX-1 upregulation and increased lipid peroxidation, HIF-1α stabilization and p38α activation. Therefore, these molecules may be considered new targets in CIH-induced RVH.

Coffee pulp is a by-product generated during coffee processing, producing environmental pollution when discarded in rivers. The revalorization of coffee by-products is currently being studied due to their high potential as new value-added food ingredients. This work aims to explore the chemical composition and functional properties of coffee pulp and validate its safety for use as a novel food ingredient. Coffee pulp composition was assessed following the standard methods (AOAC). Physicochemical properties, total phenolic compounds (TPC), antioxidant capacity, hypoglycemic and hypolipidemic properties were determined using in vitro techniques. Acute and sub-chronic oral toxicity experiments were carried out following OECD Test Guidelines 452 and 408. Coffee pulp showed a high content of dietary fiber (51.2%), highlighting the insoluble fraction. Proteins (9.2%) and lipids (2.6%) were considered a min fraction. Coffee pulp presented high content of TPC (40.6 mg g⁻¹), of which 29% were linked to DF, giving coffee pulp its antioxidant potential (102.3 mg Trolox eq g⁻¹). The by-product showed good physicochemical properties. Coffee pulp reduced the absorption of cholesterol (84%) and bile salts (85%), inhibited pancreatic lipase (64%), and presented the capacity to diminish the diffusion of glucose (34%). The intake of coffee pulp did not cause significant lesions in vital organs. In conclusion, the coffee pulp could be used as a potential ingredient with beneficial health properties.

Fetal undernutrition programs hypertension and cardiovascular diseases, and resistance artery remodeling may be a contributing factor. We aimed to assess if fetal undernutrition induces resistance artery remodeling and the relationship with hypertension. Sprague–Dawley dams were fed ad libitum (Control) or with 50% of control intake between days 11 and 21 of gestation (maternal undernutrition, MUN). In six-month-old male and female offspring we assessed blood pressure (anesthetized and tail-cuff); mesenteric resistance artery (MRA) structure and mechanics (pressure myography), cellular and internal elastic lamina (IEL) organization (confocal microscopy) and plasma MMP-2 and MMP-9 activity (zymography). Systolic blood pressure (SBP, tail-cuff) and plasma MMP activity were assessed in 18-month-old rats. At the age of six months MUN males exhibited significantly higher blood pressure (anesthetized or tail-cuff) and plasma MMP-9 activity, while MUN females did not exhibit significant differences, compared to sex-matched controls. MRA from 6-month-old MUN males and females showed a smaller diameter, reduced adventitial, smooth muscle cell density and IEL fenestra area, and a leftward shift of stress-strain curves. At the age of eighteen months SBP and MMP-9 activity were higher in both MUN males and females, compared to sex-matched controls. These data suggest that fetal undernutrition induces MRA inward eutrophic remodeling and stiffness in both sexes, independent of blood pressure level. Resistance artery structural and mechanical alterations can participate in the development of hypertension in aged females and may contribute to adverse cardiovascular events associated with low birth weight in both sexes.

Early breastfeeding cessation is a major public health problem. Several factors can affect breastfeeding pattern, and psychological aspects have been poorly explored. We hypothesize that psychological factors and breastfeeding pattern have a relationship. We have assessed in mothers during the first six months of lactation if breastfeeding pattern is associated with maternal stress, postpartum depression, and dispositional optimism, and if these psychological factors play a role on breastfeeding adherence. In total, 711 women participated, answering online the following questionnaires: sociodemographic, perceived stress scale, Edinburgh postpartum depression scale, life orientation test, and breastfeeding adherence score. Women were categorized according to infant feeding practices as exclusive breastfeeding (EBF) or mixed breastfeeding (MBF). The EBF group had a lower score of perceived stress compared to those giving MBF (first month: EBF = 1.5 [1.1; 1.9], MBF = 1.8 [1.5; 2.0]; p-Value = 0.030; third month: EBF = 1.6 [1.2; 2.0], MBF = 1.8 [1.5; 2.4]; p-Value = 0.038) and also had a lower score of postpartum depression (third month: EBF = 8.0 [6.0; 11.0], MBF = 11.0 [9.0; 15.0]; p-Value = 0.001). The breastfeeding adherence score showed a positive correlation with maternal perceived stress (first month: ρ = 0.27; p-Value = 0.018), and postpartum depression (third month: ρ = 0.30; p-Value < 0.001), and a negative correlation with maternal dispositional optimism (second month: ρ = −0.20; p-Value = 0.028). MBF was positively associated with breastfeeding adherence score (odd ratio (OR) = 1.4 [1.2–1.6]; p-Value < 0.001) and with postpartum depression (OR = 1.1 [1.0; 1.1]; p-Value = 0.020). In the third month of breastfeeding, women with MBF exhibited higher perceive stress and postpartum depression compared to those with EBF and no difference in dispositional optimism. The maternal psychological aspects are associated with breastfeeding pattern. Evaluation of maternal psychological concerns and providing support to lactating mothers may help improving breastfeeding adherence.

Breast milk (BM) is beneficial due to its content in a wide range of different antioxidants, particularly relevant for preterm infants, who are at higher risk of oxidative stress. We hypothesize that BM antioxidants are adapted to gestational age and are negatively influenced by maternal age. Fifty breastfeeding women from two hospitals (Madrid, Spain) provided BM samples at days 7, 14 and 28 of lactation to assess total antioxidant capacity (ABTS), thiol groups, reduced glutathione (GSH), superoxide dismutase (SOD) and catalase activities, lipid peroxidation (malondialdehyde, MDA + 4-Hydroxy-Trans-2-Nonenal, HNE), protein oxidation (carbonyl groups) (spectrophotometry) and melatonin (ELISA). Mixed random-effects linear regression models were used to study the influence of maternal and gestational ages on BM antioxidants, adjusted by days of lactation. Regression models evidenced a negative association between maternal age and BM melatonin levels (β = −7.4 ± 2.5; p-value = 0.005); and a negative association between gestational age and BM total antioxidant capacity (β = −0.008 ± 0.003; p-value = 0.006), SOD activity (β = −0.002 ± 0.001; p-value = 0.043) and protein oxidation (β = −0.22 ± 0.07; p-value = 0.001). In conclusion, BM antioxidants are adapted to gestational age providing higher levels to infants with lower degree of maturation; maternal ageing has a negative influence on melatonin, a key antioxidant hormone.

We aimed to design and validate a new questionnaire of adherence to healthy food pyramid (HFP) (AP-Q), to improve previous instruments. The questionnaire was self-administered and included 28 questions from 10 categories (physical activity, health habits, hydration, grains, fruits, vegetables, oil type, dairy products, animal proteins, and snacks). A population of 130 Spanish adults answered it, obtaining scores from each category and a global score of HFP adherence (AP-Q score). Validation was performed through principal components analysis (PCA) and internal consistency by Cronbach’s alpha. AP-Q was also externally validated with Kidmed-test, answered by 45 individuals from the cohort. The global AP-Q score was 5.1 ± 1.3, with an internal consistency of 64%. The PCA analysis extracted seven principal components, which explained 68.5% of the variance. The global AP-Q score was positively associated with Kidmed-test score. Our data suggest that AP-Q is a complete and robust questionnaire to assess HFP adherence, with several advantages: easy to complete, cost-effective, timesaving and has the competency to assess, besides diet, several features affecting health status, lacking in other instruments. We suggest that AP-Q could be useful in epidemiological research, although it requires additional calibration to analyze its reproducibility and validation in other populations.

Twin pregnancies are increasing due to the rise in mothers’ childbearing age and have a higher risk of fetal growth restriction (FGR) and prematurity. Therefore, early prediction of these events is important. Our aim was to analyze in the first trimester of pregnancy a possible association between antioxidants, including melatonin, in maternal plasma and the development of fetal complications in twin pregnancies. A single-center, prospective, and observational study was performed in 104 twin-pregnant women. A blood sample was extracted between the 9th and the 11th week of gestation, and plasma was obtained. Antioxidants (thiols, reduced glutathione, phenolic compounds, catalase, superoxide dismutase) and oxidative damage biomarkers (carbonyl groups and malondialdehyde) were assessed by spectrophotometry, and global scores were calculated from these parameters (Antiox-S, Prooxy-S). Melatonin and cortisol were evaluated by a competitive immunoassay. In the first trimester of pregnancy, Antiox-S was significantly lower in women who developed FGR compared to those with normal fetal growth; plasma melatonin was significantly lower in women with preterm compared to those with full-term births and exhibited a positive correlation with birth weight. Maternal cortisol showed a negative correlation with birth weight. We conclude that, for twin gestations, maternal plasma antioxidant status and melatonin could be potential biomarkers to be included in algorithms to predict FGR and preterm labor.