Magnesium and calcium content in waters from the Mediterranean coastal area of south-eastern Spain and their relation with industrial activity

Se han determinado los niveles de calcio y magnesio en aguas (potables, de riego, residuales y marinas) de la zona industrial y costera del sureste de España. Las muestras de agua acidifi cadas fueron analizadas por espectrometría de absorción atómica. Previamente las muestras fueron tratadas con 0,2 ml de LaCl3 (10 mg/mL La) como modifi cador de matriz. Las concentraciones de magnesio oscilaron entre 18,2 μg/L y 2286,7 μg/L y las de calcio desde 48,8 μg/L a 627,1 μg/L. Los niveles medios de magnesio y calcio en aguas marinas fueron signifi cativamente superiores (P < 0,001). Las concentraciones de magnesio encontradas en los diferentes grupos de aguas estuvieron linear y signifi - cativamente correlacionadas con los niveles de calcio correspondientes (r= 0,942; P < 0,001). La actividad humana e industrial no infl uyó en los niveles de magnesio y calcio en las aguas de riego y marinas de la zona. Sin embargo, los efl uentes de la fábrica de papel existente en la zona y los de origen doméstico aumentaron signifi cativamente las concentraciones de calcio en las aguas residuales.Magnesium and calcium levels in waters (potable, irrigation, waste and sea waters) from a coastal and industrial zone in south-eastern Spain, have been determined. Acidifi ed water samples were analysed by atomic absorption spectrometry. The samples were previously treated with a matrix modifi er consisting in 0.2 mL of LaCl3 (10 mg/mL La). Magnesium concentrations ranged from 18.2 μg/L to 2286.7 μg/L and those for calcium from 48.8 μg/L to 627.1 μg/L. Mean magnesium and calcium levels in sea water samples were signifi cantly higher (P < 0.001). Magnesium concentrations found in different water groups were signifi cantly and linearly related with corresponding calcium levels (r= 0.942; P < 0.001). Human and industrial activity did not infl uence magnesium and calcium levels in irrigation and sea waters of the zone. Nevertheless, effl uents from the paper factory existing in the zone and from domestic origin increased signifi cantly the calcium concentrations in waste waters

Bioaccessible peptides released by in vitro gastrointestinal digestion of fermented goat milks

In this study, ultrafiltered goat milks fermented with the classical starter bacteria Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus salivarus subsp. thermophilus or with the classical starter plus the Lactobacillus plantarum C4 probiotic strain were analyzed using ultra-high performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) and/or liquid chromatography-ion trap (LC-IT-MS/MS). Partial overlapping of the identified sequences with regard to fermentation culture was observed. Evaluation of the cleavage specificity suggested a lower proteolytic activity of the probiotic strain. Some of the potentially identified peptides had been previously reported as angiotensin converting enzyme (ACE)-inhibitory, antioxidant and antibacterial and might account for the in vitro activity previously reported for these fermented milks. Simulated digestion of the products was conducted in presence of a dialysis membrane to retrieve the bioaccessible peptide fraction. Some sequences with reported physiological activity resisted digestion but were found in the non-dialyzable fraction. However, non-previously detected sequences such as the antioxidant αs1-casein 144YFYPQL149, the antihypertensive αs2-casein 90YQKFPQY96 and the antibacterial αs2-casein 165LKKISQ170 were found in the dialyzable fraction of both fermented milks. Moreover, in the fermented milk including the probiotic strain, the k-casein dipeptidyl peptidase IV inhibitor (DPP-IV) 51INNQFLPYPY60 as well as additional ACE-inhibitory or antioxidant sequences could be identified. With the aim to anticipate further biological outcomes, quantitative structure activity relationship (QSAR) analysis was applied to the bioaccessible fragments and led to propose potential ACE inhibitory sequences

Melatonin Improves Mitochondrial Dynamics and Function in the Kidney of Zücker Diabetic Fatty Rats

Obesity and associated diabetes (diabesity) impair kidney mitochondrial dynamics by augmenting fission and diminishing fusion, which results in mitochondrial and renal dysfunction. Based on available evidence, the antioxidant activities of melatonin may improve impaired renal mitochondrial function in obese diabetic animals by restoring the imbalanced dynamics through inhibiting fission and promoting fusion. Male Zücker diabetic fatty (ZDF) rats and lean littermates (ZL) were orally treated either with melatonin (10 mg/kg BW/day) (M-ZDF and M-ZL) or vehicle (C-ZDF and C-ZL) for 17 weeks. Kidney function was evaluated by measurement of total urine volume, proteinuria, creatinine clearance, and assessment of kidney mitochondrial dynamics and function. C-ZDF exhibited impaired dynamics and function of kidney mitochondria in comparison to C-ZL. Melatonin improved nephropathy of ZDF rats and modulated their mitochondrial dynamics by reducing expression of Drp1 fission marker and increasing that of fusion markers, Mfn2 and Opa1. Furthermore, melatonin ameliorated mitochondrial dysfunction by increasing respiratory control index and electron transfer chain complex IV activity. In addition, it lowered mitochondrial oxidative status. Our findings show that melatonin supplementation improves nephropathy likely via modulation of the mitochondrial fission/fusion balance and function in ZDF rats.Ministerio de Economia y Competitividad (Spain) SAF2016-79794-

Hypotensive effect of an extract of bioactive compounds of olive leaves: preliminary clinicals study

Objetivo: evaluar el posible efecto hipotensor en sujetos pre-hipertensos e hipertensos, de un extracto de hoja de olivo (EHO) estandarizado al 15% (m/m) en oleoeuropeína y con un contenido medio del 10% (m/m) en ácidos triterpénicos y del 1% (m/m) en hidroxitirosol. Asimismo, se ha valorado su acción sobre el estatus antioxidante y el perfil lipídico sanguíneos. Material y métodos: se ha llevado a cabo un ensayo de intervención en humanos con administración de 1.600 mg de extracto/día. Los parámetros evaluados han sido presión sistólica y diastólica, retardo de la oxidación del colesterol LDL (lag time) y niveles sanguíneos de óxido nítrico (NO), malondialdehído (MDA), capacidad antioxidante (CAO), perfil lipídico, glucosa, transaminasas y creatinina. Resultados: se observó en todos los sujetos una disminución estadísticamente significativa de la presión sistólica y diastólica, y un aumento de los niveles de NO (P < 0,050). En los sujetos con presión sistólica más elevada también se redujo significativamente su valor tras la intervención (P = 0,002). El “lag time” de las LDL aumentó significativamente (P = 0,047), y en todos los sujetos los niveles de colesterol (CHO) se redujeron significativamente. Los niveles de colesterol LDL, triglicéridos (TG) y del índice CHO/colesterol HDL disminuyeron con tendencia a la significancia (P = 0,076; P = 0,059; P = 0,056; respectivamente). Conclusión: este estudio, aunque preliminar, pone de relieve por primera vez la influencia positiva del EHO ensayado en la regulación de la hipertensión así como en la velocidad de oxidación de las LDL y en el perfil lipídico sanguíneo.Objective: to evaluate the possible hypotensive effect in pre-hypertensive and hypertensive subjects of an olive leaf extract (OLE) standardized to 15% (w/w) in oleoeuropein, and with a 10% (w/w) mean content of triterpenic acids and 1% (w/w) in hydroxytirosol. Moreover, the possible effects on the blood antioxidant status and lipid profile have been also evaluated. Materials and methods: this interventional study has been performed in human volunteers, to whom 1 600 mg OLE/days, was administered. The analyzed parameters at the beginning and end of the study were diastolic and systolic pressure, delay in the LDL-cholesterol oxidation “lag time” and blood levels of nitric oxide (NO), malonic dialdehyde (MDA), antioxidant capacity (AOC) lipid profile, glucose, transaminases and creatinine. Results: a decrease in the diastolic and systolic pressure, and an increase in the NO values all statistically significant (CHO) for all volunteers, was found. In volunteers with higher systolic pressure their levels were also significantly diminished after the intervention trial (P = 0,002). The LDL lag time increased significantly (P = 0,047). Additionally, in all volunteers CHO levels were significantly decreased, and those of LDL cholesterol, triglycerides (TG) and the CHO/HDL cholesterol ratio were diminished with a tendency to the significance (P = 0,076; P = 0,059; P = 0,056; respectively). Conclusions: this preliminary study reports by the first time the positive influence of the OLE assayed in the regulation of the hypertension, LDL lag time and blood lipid profile. Therefore, further studies are of great interest.Queremos manifestar nuestro agradecimiento a la empresa Biosearch Life y al grupo AGR-141, colaboradores directos en el desarrollo y financiación de los ensayos necesarios para la realización del estudio

Total Zn of foods and bioaccesible fractions in the small and large intestine after in vitro digestion and fermentation with fecal material of healthy adults and children: Influence of culinary techniques

This work was supported by the European Research Commission (Research Executive Agency) under the research project Stance4Health under Grant Contract No 816303 and by the Plan propio de Investigación y Transferencia of the University of Granada under the program “Intensificación de la Investigación, modalidad B”.The healthy status of human beings is associated with an appropriate nutritional status in Zn, which must firstly be bioavailable. We measured the total Zn amount and its bioaccesibility in raw foods and after cooking by common culinary techniques. These foods were submitted to an in vitro digestion and fermentation with faecal inocula from healthy adults and children to evaluate Zn bioaccesibility in the small and large intestine. Mean total Zn amount provided by foods was 8.080 μg/g. Zn amount released from food in the small intestine was significantly different among several food groups and lower in raw vegetal foods compared to cooked ones (frying, roasting and grilling; p < 0.05); the same behaviour was found in the large intestine for healthy children. Zn bioaccesibility in the large intestine varied statistically according to the subjects’ idiosyncrasies, and was higher in healthy children (p < 0.05) probably due to growth demands and different composition of the colonic microbiota. In healthy adults and children, the bioaccesible fractions were 33.0 ± 20.4 % for the small intestine, 16.4 ± 22.0 and 59.6 ± 29.9% for the large one, and the non-bioaccessible ones 50.6 ± 19.9 and 7.4 ± 9.1%, respectively.European Research Commission 816303Plan propio de Investigación y Transferencia of the University of GranadaUniversidad de Granada UG

Melatonin induces fat browning by transdifferentiation of white adipocytes and de novo differentiation of mesenchymal stem cells

This research was partially supported by grant SAF2016-79794R from the Ministerio de Ciencia e Innovacion (Spain), European Regional Development Fund (ERDF), University of Granada and FEDER Funds grant number B-CTS-102-UGR20. The authors thank Antonio Tirado for their technical assistance.The role of melatonin in obesity control is extensively accepted, but its mechanism of action is still unclear. Previously we demonstrated that chronic oral melatonin acts as a brown-fat inducer, driving subcutaneous white adipose tissue (sWAT) into a brown-fat-like function (beige) in obese diabetic rats. However, immunofluorescence characterization of beige depots in sWAT and whether melatonin is a beige-fat inducer by de novo differentiation and/or transdifferentiation of white adipocytes are still undefined. Lean (ZL) and diabetic fatty (ZDF) Zücker rats were subdivided into two groups, control (C) and oral melatonin-supplemented (M, 10 mg kg−1 day−1) for 6 weeks. Mesenchymal stem cells (MSCs) were isolated from both rat inguinal fat and human lipoaspirates followed by adipogenesis assays with or without melatonin (50 nM for 12 h in a 24 h period, 12 h+/12 h−) mimicking the light/dark cycle. Immunofluorescence and western-blot assays showed the partial transdifferentiation of white adipocytes in both ZL and ZDF rats, with increasing thermogenic and beige markers, UCP1 and CITED1 and decreasing white adipocyte marker ASC-1 expression. In addition, melatonin increased UCP1, CITED1, and PGC1- α expression in differentiated adipocytes in both rats and humans. These results demonstrate that melatonin increases brown fat in obese diabetic rats by both adipocyte transdifferentiation and de novo differentiation. Furthermore, it promotes beige MSC adipogenesis in humans. This may contribute to the control of body weight attributed to melatonin and its metabolic benefits in human diabesity.Spanish Government SAF2016-79794REuropean Commission B-CTS-102-UGR20University of Granad

Antioxidant, ACE-inhibitory and antimicrobial activity of fermented goat milk: activity and physicochemical property relationship of the peptide components

Increasing evidence on goat milk and the health benefits of its derived products beyond its nutritional value show its potential as a functional food. In this study, goat milk fractions were tested for their total antioxidant capacity using different methods (ORAC, ABTS, DPPH and FRAP), as well as their angiotensin-I-converting-enzyme inhibitory and antimicrobial (against Escherichia coli and Micrococcus luteus) activities. Different whey fractions (whey, cation exchange membrane permeate P and retentate R) of two fermented skimmed goat milks (ultrafiltered goat milk fermented with the classical starter bacteria or with the classical starter plus the Lactobacillus plantarum C4 probiotic strain) were assessed. Additionally, P fractions were divided into two sub-fractions after being passed through a 3 kDa cut-off membrane: (a) the permeate with peptides of MW 3 kDa (P > 3). No differences in biological activities were observed between the two fermented milks. However, the biological peptides present in the P < 3 fraction showed the highest total antioxidant capacity (for the ORAC assay) and angiotensin-I-converting-enzyme inhibitory activity. Those present in the R fraction showed the highest total antioxidant capacity against ABTS˙+ and DPPH˙ radicals. Some antimicrobial activity against E. coli was observed for the fermented milk containing the probiotic, which could be due to some peptides being released by the probiotic strain. In conclusion, small and non-basic bioactive peptides could be responsible for most of the angiotensin-I-converting-enzyme inhibitory and antioxidant activities. These findings reinforce the potential benefits of the consumption of fermented goat milk in the prevention of cardiovascular diseases associated with oxidative stress and hypertension

Hydrochars Derived from Spent Coffee Grounds as Zn Bio-Chelates for Agronomic Biofortification

Previous studies have attributed both phytotoxicity and the capacity to mobilize nutrient elements to the presence of polyphenols and melanoidins in spent coffee grounds (SCG) and SCG-hydrochars obtained through hydrothermal carbonization (HTC). This work aimed to evaluate SCG and two SCG-hydrochars obtained at 160 and 200 °C that were functionalized with Zn salts (bio-chelates), to achieve the in vitro biofortification of lettuce. Two application modes were established: (1) a fixed Zn concentration of 10 mg kg−1 of soil and (2) a fixed dose of 0.5% bio-product. Soil alone (control A) and commercial chelates (control B) were used as controls. Outcomes showed that SCG-hydrochars retain the capacity to mobilize Zn compared to SCG. However, the chelating capacity was reduced (Zn: 94%) and the toxicity was significantly increased (p < 0.05) with higher temperatures of HTC (200 °C). Both fresh and dry lettuce weights were less affected at doses of 0.5% of bio-product and registered a maximum increase of 136% of Zn in the plant content. The present study approaches the possibility of using these by-products as bioinorganic fertilizers at subtoxic doses, although more research is neededPY20_00585 from FEDER and RDPTC-2018 (AT17_6096_OTRI UGR) from the Andalusia n Ministry of Economic Transformation, Knowledge, Industry and Universities

Melatonin Enhances the Mitochondrial Functionality of Brown Adipose Tissue in Obese—Diabetic Rats

Developing novel drugs/targets remains a major effort toward controlling obesity-related type 2 diabetes (diabesity). Melatonin controls obesity and improves glucose homeostasis in rodents, mainly via the thermogenic effects of increasing the amount of brown adipose tissue (BAT) and increases in mitochondrial mass, amount of UCP1 protein, and thermogenic capacity. Importantly, mitochondria are widely known as a therapeutic target of melatonin; however, direct evidence of melatonin on the function of mitochondria from BAT and the mechanistic pathways underlying these effects remains lacking. This study investigated the effects of melatonin on mitochondrial functions in BAT of Zücker diabetic fatty (ZDF) rats, which are considered a model of obesity-related type 2 diabetes mellitus (T2DM). At five weeks of age, Zücker lean (ZL) and ZDF rats were subdivided into two groups, consisting of control and treated with oral melatonin for six weeks. Mitochondria were isolated from BAT of animals from both groups, using subcellular fractionation techniques, followed by measurement of several mitochondrial parameters, including respiratory control ratio (RCR), phosphorylation coefficient (ADP/O ratio), ATP production, level of mitochondrial nitrites, superoxide dismutase activity, and alteration in the mitochondrial permeability transition pore (mPTP). Interestingly, melatonin increased RCR in mitochondria from brown fat of both ZL and ZDF rats through the reduction of the proton leak component of respiration (state 4). In addition, melatonin improved the ADP/O ratio in obese rats and augmented ATP production in lean rats. Further, melatonin reduced mitochondrial nitrosative and oxidative status by decreasing nitrite levels and increasing superoxide dismutase activity in both groups, as well as inhibited mPTP in mitochondria isolated from brown fat. Taken together, the present data revealed that chronic oral administration of melatonin improved mitochondrial respiration in brown adipocytes, while decreasing oxidative and nitrosative stress and susceptibility of adipocytes to apoptosis in ZDF rats, suggesting a beneficial use in the treatment of diabesity. Further research regarding the molecular mechanisms underlying the effects of melatonin on diabesity is warranted.SAF2016-79794-R from the Ministerio de Ciencia e Innovación (Spain)European Regional Development Fund (ERDF

Melatonin Improves Levels of Zn and Cu in the Muscle of Diabetic Obese Rats

Melatonin improves metabolic alterations associated with obesity and its diabetes (diabesity). We intend to determine whether this improvement is exerted by changing Zn and/or Cu tissue levels in liver, muscle, pancreas, and brain, and in internal (perirenal, perigonadal, and omentum) and subcutaneous lumbar white adipose tissues (IWAT and SWAT, respectively). Male Zücker diabetic fatty (ZDF) rats and lean littermates (ZL) were orally supplemented either with melatonin (10 mg/kg body weight/day) or vehicle for 6 weeks. Zn and Cu concentrations were not significantly influenced by diabesity in the analyzed tissues (p > 0.05), with the exception of Zn in liver. In skeletal muscle Zn and Cu, and in perirenal WAT, only Zn levels increased significantly with melatonin supplementation in ZDF rats (p < 0.05). This cytoplasmic Zn enhancement would be probably associated with the upregulation of several Zn influx membrane transporters (Zips) and could explain the amelioration in the glycaemia and insulinaemia by upregulating the Akt and downregulating the inhibitor PTP1B, in obese and diabetic conditions. Enhanced Zn and Cu levels in muscle cells could be related to the reported antioxidant melatonin activity exerted by increasing the Zn, Cu-SOD, and extracellular Cu-SOD activity. In conclusion, melatonin, by increasing the muscle levels of Zn and Cu, joined with our previously reported findings improves glycaemia, insulinaemia, and oxidative stress in this diabesity animal model.SAF2016-79794-R from the Ministerio de Economía y Competitividad (Spain) and European Regional Development Fund (ERDF)