Synergistic Bactericidal Effects of Quaternary Ammonium Compounds with Essential Oil Constituents.

Antimicrobial tolerance is a significant concern in the food industry, as it poses risks to food safety and public health. To overcome this challenge, synergistic combinations of antimicrobials have emerged as a potential solution. In this study, the combinations of two essential oil constituents (EOCs), namely carvacrol (CAR) and eugenol (EUG), with the quaternary ammonium compounds (QACs) benzalkonium chloride (BAC) and didecyldimethylammonium chloride (DDAC) were evaluated for their antimicrobial effects against Escherichia coli and Bacillus cereus, two common foodborne bacteria. The checkerboard assay was employed to determine the fractional inhibitory concentration index (FICI) and the fractional bactericidal concentration index (FBCI), indicating the presence of bactericidal, but not bacteriostatic, synergy in all QAC-EOC combinations. Bactericidal synergism was clearly supported by Bliss independence analysis. The bactericidal activity of the promising synergistic combinations was further validated by time-kill curves, achieving a >4-log10 reduction of initial bacterial load, which is significant compared to typical industry standards. The combinations containing DDAC showed the highest efficiency, resulting in the eradication of bacterial population in less than 2-4 h. These findings emphasize the importance of considering both bacteriostatic and bactericidal effects when evaluating antimicrobial combinations and the potential of EOC-QAC combinations for sanitization and disinfection in the food industry.

Valorisation of pork by-products to obtain antioxidant and antihypertensive peptides.

The porcine liver could be used for the extraction of zinc-protoporphyrin (ZnPP) as a natural red meat pigment. During the autolysis process, porcine liver homogenates was incubated at pH 4.8 and 45 °C under anaerobic conditions to obtain insoluble ZnPP. After incubation, the homogenates were readjusted at pH 4.8, and at pH 7.5 before being centrifuged at 5500 × g for 20 min at 4 °C and the resulting supernatant were compared with the obtained at pH 4.8 at the beginning of the incubation. The molecular weight distributions of the porcine liver fractions at both pHs were very similar, however, eight essential amino acids were more abundant in fractions obtained at pH 4.8. Regarding the ORAC assay, porcine liver protein fraction at pH 4.8 showed the highest antioxidant capacity but antihypertensive inhibition was similar for both pHs. Peptides with strong bioactivity potential from aldehyde dehydrogenase, lactoylglutathione lyase, SEC14-like protein 3 and others were identified. The findings have demonstrated the potential of the porcine liver to extract natural pigments and bioactive peptides.

Hyaluronan Brush-like Copolymers Promote CD44 Declustering in Breast Cancer Cells.

We report on the synthesis of hyaluronan (HA) brush-like copolymers and their application as antagonists of tumorigenic CD44-HA interactions. HA (4.8 kDa, ca. 24 saccharides) was grafted on 2-hydrohyethyl methacrylate (HEMA) by end-on oxime ligation. The obtained copolymers were compared with low and high molecular weight HA in terms of hydrolysis kinetics in the presence of hyaluronidase (isothermal titration calorimetry) and interactions with CD44 (surface plasmon resonance). The results evidenced that the high molecular weight HA and HA-g-HEMA have a much higher affinity to CD44 than low molecular weight HA. Additionally, slower enzymatic degradation was observed for the copolymer, making it an excellent candidate for active targeting of tumorigenic CD44-HA interactions. We, therefore, investigated the effect of the copolymer on cancer cell lines with different expression of CD44 and observed an efficient declustering of CD44 that is usually associated with reduction of metastasis and drug resistance.

Ionogels Derived from Fluorinated Ionic Liquids to Enhance Aqueous Drug Solubility for Local Drug Administration.

Gelatin is a popular biopolymer for biomedical applications due to its harmless impact with a negligible inflammatory response in the host organism. Gelatin interacts with soluble molecules in aqueous media as ionic counterparts such as ionic liquids (ILs) to be used as cosolvents to generate the so-called Ionogels. The perfluorinated IL (FIL), 1-ethyl-3-methylpyridinium perfluorobutanesulfonate, has been selected as co-hydrosolvent for fish gelatin due to its low cytotoxicity and hydrophobicity aprotic polar structure to improve the drug aqueous solubility. A series of FIL/water emulsions with different FIL content and their corresponding shark gelatin/FIL Ionogel has been designed to enhance the drug solubility whilst retaining the mechanical structure and their nanostructure was probed by simultaneous SAXS/WAXS, FTIR and Raman spectroscopy, DSC and rheological experiments. Likewise, the FIL assisted the solubility of the antitumoural Doxorubicin whilst retaining the performing mechanical properties of the drug delivery system network for the drug storage as well as the local administration by a syringe. In addition, the different controlled release mechanisms of two different antitumoral such as Doxorubicin and Mithramycin from two different Ionogels formulations were compared to previous gelatin hydrogels which proved the key structure correlation required to attain specific therapeutic dosages.

Characterization of Tuna Gelatin-Based Hydrogels as a Matrix for Drug Delivery.

The skin of yellowfin tuna is one of the fishery industry solid residues with the greatest potential to add extra value to its circular economy that remains yet unexploited. Particularly, the high collagen content of fish skin allows generating gelatin by hydrolysis, which is ideal for forming hydrogels due to its biocompatibility and gelling capability. Hydrogels have been used as drug carriers for local administration due to their mechanical properties and drug loading capacity. Herein, novel tuna gelatin hydrogels were designed as drug vehicles with two structurally different antitumoral model compounds such as Doxorubicin and Crocin to be administrated locally in tissues with complex human anatomies after surgical resection. The characterization by gel permeation chromatography (GPC) of purified gelatin confirmed their heterogeneity composition, exhibiting three major bands that correspond to the ß and α chains along with high molecular weight species. In addition, the Fourier Transform Infrared (FT-IR) spectra of gelatin probed the secondary structure of the gelatin showing the simultaneous existence of α helix, ß sheet, and random coil structures. Morphological studies at different length scales were performed by a multi-technique approach using SAXS/WAXS, AFM and cryo-SEM that revealed the porous network formed by the interaction of gelatin planar aggregates. In addition, the sol-gel transition, as well as the gelation point and the hydrogel strength, were studied using dynamic rheology and differential scanning calorimetry. Likewise, the loading and release profiles followed by UV-visible spectroscopy indicated that the novel gelatin hydrogels improve the drug release of Doxorubicin and Crocin in a sustained fashion, indicating the structure-function importance in the material composition.

Deciphering Structural Determinants in Chondroitin Sulfate Binding to FGF-2: Paving the Way to Enhanced Predictability of their Biological Functions.

Controlling chondroitin sulfates (CSs) biological functions to exploit their interesting potential biomedical applications requires a comprehensive understanding of how the specific sulfate distribution along the polysaccharide backbone can impact in their biological activities, a still challenging issue. To this aim, herein, we have applied an "holistic approach" recently developed by us to look globally how a specific sulfate distribution within CS disaccharide epitopes can direct the binding of these polysaccharides to growth factors. To do this, we have analyzed several polysaccharides of marine origin and semi-synthetic polysaccharides, the latter to isolate the structure-activity relationships of their rare, and even unnatural, sulfated disaccharide epitopes. SPR studies revealed that all the tested polysaccharides bind to FGF-2 (with exception of CS-8, CS-12 and CS-13) according to a model in which the CSs first form a weak complex with the protein, which is followed by maturation to tight binding with k D ranging affinities from ~ 1.31 µM to 130 µM for the first step and from ~ 3.88 µM to 1.8 nM for the second one. These binding capacities are, interestingly, related with the surface charge of the 3D-structure that is modulated by the particular sulfate distribution within the disaccharide repeating-units.

Production, Characterization, and Bioactivity of Fish Protein Hydrolysates from Aquaculture Turbot (Scophthalmus maximus) Wastes.

The valorization of wastes generated in the processing of farmed fish is currently an issue of extreme relevance for the industry, aiming to accomplish the objectives of circular bioeconomy. In the present report, turbot (Scophthalmus maximus) by-products were subjected to Alcalase hydrolysis under the optimal conditions initially defined by response surface methodology. All the fish protein hydrolysates (FPHs) showed a high yield of digestion (>83%), very remarkable degrees of hydrolysis (30-37%), high content of soluble protein (>62 g/L), an excellent profile of amino acids, and almost total in vitro digestibility (higher than 92%). Antioxidant and antihypertensive activities were analyzed in all cases, viscera hydrolysates being the most active. The range of average molecular weights (Mw) of turbot hydrolysates varied from 1200 to 1669 Da, and peptide size distribution showed that the hydrolysate of viscera had the highest content of peptides above 1000 Da and below 200 Da.

Impact of Prevalence Ratios of Chondroitin Sulfate (CS)- 4 and -6 Isomers Derived from Marine Sources in Cell Proliferation and Chondrogenic Differentiation Processes.

Osteoarthritis is the most prevalent rheumatic disease. During disease progression, differences have been described in the prevalence of chondroitin sulfate (CS) isomers. Marine derived-CS present a higher proportion of the 6S isomer, offering therapeutic potential. Accordingly, we evaluated the effect of exogenous supplementation of CS, derived from the small spotted catshark (Scyliorhinus canicula), blue shark (Prionace glauca), thornback skate (Raja clavata) and bovine CS (reference), on the proliferation of osteochondral cell lines (MG-63 and T/C-28a2) and the chondrogenic differentiation of mesenchymal stromal cells (MSCs). MG-G3 proliferation was comparable between R. clavata (CS-6 intermediate ratio) and bovine CS (CS-4 enrichment), for concentrations below 0.5 mg/mL, defined as a toxicity threshold. T/C-28a2 proliferation was significantly improved by intermediate ratios of CS-6 and -4 isomers (S. canicula and R. clavata). A dose-dependent response was observed for S. canicula (200 µg/mL vs 50 and 10 µg/mL) and bovine CS (200 and 100 µg/mL vs 10 µg/mL). CS sulfation patterns discretely affected MSCs chondrogenesis; even though S. canicula and R. clavata CS up-regulated chondrogenic markers expression (aggrecan and collagen type II) these were not statistically significant. We demonstrate that intermediate values of CS-4 and -6 isomers improve cell proliferation and offer potential for chondrogenic promotion, although more studies are needed to elucidate its mechanism of action.

Respuestas evocadas visuales mediante estimulación con diodos emisores de luz montados en gafas / Visual evoked responses to light-emitting diodes in goggles

Introducción: El potencial evocado visual es una técnica que permite la evaluación de la integridad funcional de la vía visual; su aplicación en niños pequeños resulta de gran dificultad. Objetivo: Caracterizar desde el punto de vista fisiológico el potencial evocado visual en niños menores de 5 años. Métodos: Estudio descriptivo transversal, donde se registran respuestas visuales obtenidas con diodos (montados en gafas) en 112 niños sanos (65 varones, 47 hembras) con rango de edad entre 3 meses y 5 años. Se calcularon los valores medios y desviación estándar de latencias y amplitudes de cada componente de la respuesta y se estimaron sus dinámicas madurativas en función de la edad. Se utilizó un análisis de componentes principales para explicar la variabilidad morfológica (complejo N1-P1-N2) y, se calculó una ecuación discriminante (con indicadores de replicación y relación señal/ruido), para evaluar la contribución en su mejor detección. Resultados: La morfología típica encontrada resultó consistente, replicable y mostró un nivel de detección visual > 90 por ciento (para cada componente), con 85 por ciento de acierto global (clasificaciones correctas) mediante la ecuación discriminante. El complejo N1-P1-N2, consigue explicar > 70 por ciento de la varianza de esta respuesta. Se demostró un acortamiento significativo de latencia con la edad (regresión, p≤ 0,05) para los componentes N1, P1 (marginal, N2). Los valores de amplitud resultaron de mayor dispersión y su tendencia al incremento no alcanzó significación. Conclusiones: El registro del potencial evocado visual mediante diodos/gafas demuestra la obtención de respuestas fisiológicas consistentes y replicables (alta detectabilidad) con una variabilidad morfológica controlable(AU)

Introduction: The Visual Evoked Potentials is a neurophysiological technique to provide an objective assessment of the functional integrity of the visual pathway. However, the young children population is a particularly difficult (uncooperative) group to obtain visual responses. Objective: The purpose of this study was to describe electrophysiological characteristics of the visual response in children. Methods: Descriptive, cross-sectional study. Transient visual evoked potentials with goggles were recorded from 112 healthy children (65 males, 47 females) aged between 3 months to 5 years old. The mean (and standard deviations) for latencies and amplitudes of each component was calculated and both statistical significance in function of age was analyzed (linear regression model). Principal component analysis is use to explain the visual waveform variability. A discriminant equation (with indicators of reliability and noise/signal ratio) is calculate to evaluate contribution in detectability. Results: The typical waveform of (N1, P1, N2, components) is consistent and reliable and showed a visual detection level >90 percent (for each component) and 85 percent of global success (good classifications) with the discriminant equation. N1-P1-N2 complex is able to explain >70 percent of the visual response variance. A significant reduction of N1, P1 (N2, marginal) latencies with increasing age is demonstrated (linear regression, p≤0.05). There was no significant difference for age-dependent increased pattern of amplitude data (high variability). Conclusions: The visual evoked potentials/goggles obtained in infants and young children are consistent and reliable physiological responses (high detectability) with recognizable morphological variability(AU)

Respuestas evocadas auditivas obtenidas mediante un estímulo chirp de banda ancha / Auditory evoked responses obtained by a broad-band chirp stimulus

Introducción: La estimación de la audición con Potenciales Evocados Auditivos de Tallo Cerebral obtenidos mediante estímulos tipo chirp constituye una alternativa de reciente aplicación. Varios autores han demostrado que, este tipo de estimulación compensa el retardo de la onda sonora en la codificación de frecuencias, generándose Potenciales Evocados Auditivos de Tallo Cerebral con componentes de amplitudes mayores. Objetivos: Diseñar y generar un estímulo chirp (banda ancha) para implementar en el sistema AUDIX (La Habana, Neuronic SA) y, realizar una serie de controles para evaluar su viabilidad en el registro de Potenciales Evocados Auditivos de Tallo Cerebral en sujetos con audición normal. Métodos: Las formulaciones que se utilizaron en la generación del estímulo fueron implementadas como una función en Matlab®, y luego, insertada en el sistema AUDIX con las siguientes especificaciones técnicas, frecuencia de muestreo: 48 kHz, composición de frecuencias (ascendente): 350-11300 Hz, y duración total: 4,95 ms. Se obtuvieron registros de PEATC mediante estímulos chirp y click a un nivel de intensidad fija (60 dB nHL) en 9 sujetos adultos (18 oídos) con audición normal. Resultados: El estímulo chirp diseñado tuvo un comportamiento funcional similar a lo reportado en la literatura. Cuando se compara con los Potenciales Evocados Auditivos de Tallo Cerebral -click, la onda V de Potenciales Evocados Auditivos de Tallo Cerebral -chirp mostró valores de amplitud significativamente mayores (relación de amplitud chirp/click: 1,62), con una ganancia promedio de 54 por ciento (p< 0,001, n= 18, prueba de rangos de Wilcoxon). Conclusiones: El estímulo chirp (banda ancha) diseñado resulta más eficiente que el estímulo click para obtener registros de Potenciales Evocados Auditivos de Tallo Cerebral. Con respecto a la amplitud de la onda V, el sistema muestra un funcionamiento lineal (mejor sincronía neural). Este tipo de estimulación pudiera resultar de mucha utilidad en programas de pesquisa auditiva neonatal pues la obtención de una onda V de mayor amplitud permitiría su fácil y rápida detección, y posible automatización(AU)

Introduction: Estimation of audition through brainstem auditory evoked potentials obtained by chirp stimuli is an alternative of recent application. It has been shown by several authors that this type of stimulation compensates for retardation of the sound wave in the coding of frequencies, generating auditory evoked responses with components of higher amplitudes. Objectives: Design and develop a broad-band chirp stimulus to be implemented in the AUDIX system and conduct a control series evaluation of its viability to register brainstem auditory evoked potentials in normal-hearing subjects. Methods: The formulations used to generate the stimulus were implemented as a function on Matlab® and then incorporated into the AUDIX system with the following technical specifications: sampling frequency: 48 kHz, frequency composition (rising): 350-11 300 Hz, and total duration: 4.95 ms. BAEP registries were obtained with chirp and click stimuli at a fixed intensity level (60 dB nHL) in nine normal-hearing adult subjects (18 ears). Results: The chirp stimulus designed had a functional behavior similar to the one reported in the literature. When compared with the click-BAEP, the V wave of chirp-BAEP displayed significantly higher amplitude values (chirp/click amplitude ratio: 1.62), with an average gain of 54 percent (p< 0.001, n= 18, Wilcoxon rank test). Conclusions: The broad band chirp stimulus designed proved to be more efficient than the click stimulus to obtain registries of Brainstem Auditory Evoked Potentials. Regarding V wave amplitude, the system was found to function linearly (better neural synchrony). This type of stimulation could be very useful in neonatal hearing screening programs, since a higher amplitude V wave could facilitate its fast and easy detection and possible automation(AU)

Enzymatic purification of glucomannan from Amorphophallus oncophyllus using α-Amylase / Purificação enzimática de glucomanan de amorphophallus oncophyllus usando α -amilasas

The international standards for top glucomannan flour require a minimum of 70% glucomannan. The glucomannan content of Amorphophallus oncophyllus flour was approximately 60%, with starch as the major impurity. Elimination of starch was expected to increase the purity of glucomannan. The purpose of this research was to study starch hydrolysis of the flour using α-amylase. Temperature (35.5-84.5oC), time (0.4-3.6 h) and pH (2.2-8.8) of hydrolysis were selected as independent variables. A central composite design of response surface methodology (RSM) was performed to obtain the optimum condition. This approach was a novelty of this enzymatic purification of A. oncophyllus. Glucomannan content, starch content, and solubility were chosen as the response variables. The models were reliable for predicting the responses (R2≥ 0.771). It was predicted that the highest glucomannan content (93.0%) obtained at the lowest starch content (1.14%), which hydrolysed at pH 6.17, 84.5oC and 3.6 h. Prior the verification of the optimum hydrolysed condition from the model, the glucomannan and starch content was 81.59% and 2.27%, respectively. After purification, the absorbance of the ß-1,4 glycosidic bond increased as a sign of higher glucomannan purity. A less rough surface and irregular shape of the grain morphology was observed after purification.

Os padrões internacionais para a farinha de alta calidade de glucomanan requerem um mínimo de 70% de glucomanan. O conteúdo de glucomanano da farinha de Amorphophallus oncophyllus foi de aproximadamente 60%, com o amido como a maior impureza. Esperava-se que a eliminação do amido aumentasse a pureza do glucomanan. O objetivo desta pesquisa foi estudar a hidrólise do amido da farinha usando α-amilase. A temperature (35,5-84,5oC), o tempo (0,4-3,6 h) e o pH (2,2-8,8) da hidrólise foram selecionados como variáveis independentes. Um desenho central composto pertencente á metodologia da superfície de resposta (MSR) foi realizado para obter a condição ótima. Esta abordagem foi uma novidade desta purificação enzimática de A. oncophyllus. O conteúdo de glucomanan, conteúdo de amido e solubilidade foram escolhidos como as respostas. Os modelos foram confiáveis para predizer as respostas (R2≥ 0,771). Os modelos indicaram que o maior conteúdo de glucomanan (93,0%) foram obtidos no menor conteúdo de amido (1,14%),que hidrolisou a um pH 6,17, 84,5ºC e 3,6 h. Antes da verificação da condição hidrolisada ótima do modelo, o conteúdo de glucomanan e amido foi de 81,59% e 2,27%, respectivamente. Após a purificação, a absorbância da ligação ß-1,4 glicosídica aumentou com um sinal de maior pureza de glucomanan. Uma superfície mais lisa e forma irregular da morfologia do grão foi observada após a purificação.

Isolation and Chemical Characterization of Chondroitin Sulfate from Cartilage By-Products of Blackmouth Catshark (Galeus melastomus).

Chondroitin sulfate (CS) is a glycosaminoglycan actively researched for pharmaceutical, nutraceutical and tissue engineering applications. CS extracted from marine animals displays different features from common terrestrial sources, resulting in distinct properties, such as anti-viral and anti-metastatic. Therefore, exploration of undescribed marine species holds potential to expand the possibilities of currently-known CS. Accordingly, we have studied for the first time the production and characterization of CS from blackmouth catshark (Galeus melastomus), a shark species commonly discarded as by-catch. The process of CS purification consists of cartilage hydrolysis with alcalase, followed by two different chemical treatments and ending with membrane purification. All steps were optimized by response surface methodology. According to this, the best conditions for cartilage proteolysis were established at 52.9 °C and pH = 7.31. Subsequent purification by either alkaline treatment or hydroalcoholic alkaline precipitation yielded CS with purities of 81.2%, 82.3% and 97.4% respectively, after 30-kDa membrane separation. The molecular weight of CS obtained ranges 53⁻66 kDa, depending on the conditions. Sulfation profiles were similar for all materials, with dominant CS-C (GlcA-GalNAc6S) units (55%), followed by 23⁻24% of CS-A (GlcA-GalNAc4S), a substantial amount (15⁻16%) of CS-D (GlcA2S-GalNAc6S) and less than 7% of other disulfated and unsulfated disaccharides.

Glycosaminoglycans from marine sources as therapeutic agents.

Glycosaminoglycans (GAGs) in marine animals are different to those of terrestrial organisms, mainly in terms of molecular weight and sulfation. The therapeutic properties of GAGs are related to their ability to interact with proteins, which is very much influenced by sulfation position and patterns. Since currently GAGs cannot be chemically synthesized, they are sourced from natural products, with high intra- but also inter-species variability, in terms of chain length, disaccharide composition and sulfation pattern. Consequently, sulfated GAGs are the most interesting molecules in the marine environment and constitute the focus of the present review. In particular, chondroitin sulfate (CS) appears as the most promising compound. CS-E chains [GlcA-GalNAc(4S,6S)] extracted from squid possess antiviral and anti-metastatic activities and seem to impart signalling properties and improve the mechanical performance of cartilage engineering constructs; Squid CS-E and octopus CS-K [GlcA(3S)-GalNAc(4S)], dermatan sulfate (DS) from sea squirts [-iK units, IdoA(3S)-GalNAc(4S)] and sea urchins [-iE units, IdoA-GalNAc(4S,6S)] and hybrids CS/DS from sharks (-B/iB [GlcA/IdoA(2S)-GalNAc(4S)], -D/iD [GlcA/IdoA(2S)-GalNAc(6S)] and -E/iE units [GlcA/IdoA-GalNAc(4S,6S)]) promote neurite outgrowth and could be valuable materials for nerve regeneration. Also displaying antiviral and anti-metastatic properties, a rare CS with fucosylated branches isolated from sea cucumbers is an anticoagulant and anti-inflammatory agent. In this same line, marine heparin extracted from shrimp and sea squirt has proven anti-inflammatory properties, with the added advantage of decreased risk of bleeding because of its low anticoagulant activity.

Hydrolysates of Fish Skin Collagen: An Opportunity for Valorizing Fish Industry Byproducts.

During fish processing operations, such as skinning and filleting, the removal of collagen-containing materials can account for up to 30% of the total fish byproducts. Collagen is the main structural protein in skin, representing up to 70% of dry weight depending on the species, age and season. It has a wide range of applications including cosmetic, pharmaceutical, food industry, and medical. In the present work, collagen was obtained by pepsin extraction from the skin of two species of teleost and two species of chondrychtyes with yields varying between 14.16% and 61.17%. The storage conditions of the skins appear to influence these collagen extractions yields. Pepsin soluble collagen (PSC) was enzymatically hydrolyzed and the resultant hydrolysates were ultrafiltrated and characterized. Electrophoretic patterns showed the typical composition of type I collagen, with denaturation temperatures ranged between 23 °C and 33 °C. In terms of antioxidant capacity, results revealed significant intraspecific differences between hydrolysates, retentate, and permeate fractions when using ß-Carotene and DPPH methods and also showed interspecies differences between those fractions when using DPPH and ABTS methods. Under controlled conditions, PSC hydrolysates from Prionace glauca, Scyliorhinus canicula, Xiphias gladius, and Thunnus albacares provide a valuable source of peptides with antioxidant capacities constituting a feasible way to efficiently upgrade fish skin biomass.

Ambient temperature and prevalence of obesity in the Spanish population: The Di@bet.es study.

OBJECTIVE: The aim of this study was to examine possible associations between ambient temperature and obesity in the Spanish population using an ecological focus. METHODS: The Di@bet.es study is a national, cross-sectional, population-based survey of cardiometabolic risk factors and their association with lifestyle. SAMPLE: 5,061 subjects in 100 clusters. VARIABLES: Clinical, demographic and lifestyle survey, physical examination, and blood sampling. The mean annual temperature (°C) for each study site was collected from the Spanish National Meteorology Agency (1971-2000). RESULTS: The prevalence rates of obesity in the different geographical areas divided according to mean annual temperature quartiles were 26.9% in quartile 1 (10.4-14.5°C), 30.5% in quartile 2 (14.5-15.5°C), 32% in quartile 3 (15.5-17.8°C), and 33.6% in quartile 4 (17.8-21.3°C) (P = 0.003). Logistic regression analyses including multiple socio-demographic (age, gender, educational level, marital status) and lifestyle (physical activity, Mediterranean diet score, smoking) variables showed that, as compared with quartile 1, the odd ratios for obesity were 1.20 (1.01-1.42), 1.35 (1.12-1.61), and 1.38 (1.14-1.67) in quartiles 2, 3, and 4, respectively (P = 0.001 for difference, P < 0.001 for trend). CONCLUSIONS: Our study reports an association between ambient temperature and obesity in the Spanish population controlled for known confounders.

Optimisation of antioxidant extraction from Solanum tuberosum potato peel waste by surface response methodology.

This study reports the optimised conditions (temperature, ethanol concentration and processing-time) for antioxidant extraction from potato peel (Agria variety) waste. At short extraction times (34 min), optimal yields of phenolic (TP) and flavonoid (Fv) compounds were reached at 89.9°C and ethanol concentrations of 71.2% and 38.6%, respectively. The main phenolic compounds identified in the extracts were chlorogenic (Cl) and ferulic (Fer) acids. A significant positive correlation was found between antioxidant activity and TP, Fv, Fer and Cl responses. Potato peel extracts were able to stabilize soybean oil under accelerated oxidation conditions, minimising peroxide, totox and p-anisidine indices. The production of hexanal and 2-hexenal in soybean oil samples was maximal for extracts obtained at intermediate temperatures and ethanol concentrations. Our results demonstrate potato peel waste is a good source of antioxidants able to effectively limit oil oxidation, while contributing to the revalorisation of these agrifood by-products.

Variable patterns of obesity and cardiometabolic phenotypes and their association with lifestyle factors in the Di@bet.es study.

BACKGROUND AND AIM: Prevalence rates of "metabolically healthy obese" (MHO) subjects vary depending on the criteria used. This study examined the prevalence and characteristics of MHO subjects and metabolically abnormal normal-weight subjects and compared the findings with the NHANES 1999-2004 study. The aims of the present study were, first, to determine the prevalence rates of MHO and MNHNO subjects using the same criteria as those of the National Health and Nutrition Examination Survey (NHANES) (1999-2004) study, and second to compare the prevalence and correlates of obese subjects who are resistant to the development of adiposity-associated cardiometabolic abnormalities (CA) and normal-weight individuals who display cardiometabolic risk factor clustering between the Spanish and the US populations. METHODS AND RESULTS: Di@bet.es study is a national, cross-sectional population-based survey of 5728 adults conducted in 2009-2010. Clinical, metabolic, sociodemographic, and anthropometric data and information about lifestyle habits, such as physical activity, smoking habit, alcohol intake and food consumption, were collected. Subjects were classified according to their body mass index (BMI) (normal-weight, <25 kg/m(2); overweight, 25-29.9 kg/m(2); and obese, >30 kg/m(2)). CA included elevated blood pressure; elevated levels of triglycerides, fasting glucose, and high-sensitivity C-reactive protein (hs-CRP); and elevated homeostasis model assessment of insulin resistance (HOMA-IR) value and low high-density lipoprotein cholesterol (HDL-c) level. Two phenotypes were defined: metabolically healthy phenotype (0-1 CA) and metabolically abnormal phenotype (≥2 CA). The prevalence of metabolically abnormal normal-weight phenotype was slightly lower in the Spanish population (6.5% vs. 8.1%). The prevalence of metabolically healthy overweight and MHO subjects was 20.9% and 7.0%, respectively, while in NHANES study it was 17.9% and 9.7%, respectively. Cigarette smoking was associated with CA in each phenotype, while moderate physical activity and moderate alcohol intake were associated with being metabolically healthy. Olive oil intake was negatively associated with the prevalence of CA. CONCLUSIONS: Smoking, physical activity level, and alcohol intake contribute to the explanation of the prevalence of CA in the Spanish population, as in the US population. However in Spain, olive oil intake contributes significantly to the explanation of the variance in the prevalence of CA.

Identification of the major ACE-inhibitory peptides produced by enzymatic hydrolysis of a protein concentrate from cuttlefish wastewater.

The aim of this work was the purification and identification of the major angiotensin converting enzyme (ACE) inhibitory peptides produced by enzymatic hydrolysis of a protein concentrate recovered from a cuttlefish industrial manufacturing effluent. This process consisted on the ultrafiltration of cuttlefish softening wastewater, with a 10 kDa cut-off membrane, followed by the hydrolysis with alcalase of the retained fraction. Alcalase produced ACE inhibitors reaching the highest activity (IC50 = 76.8 ± 15.2 µg mL⁻¹) after 8 h of proteolysis. Sequential ultrafiltration of the 8 h hydrolysate with molecular weight cut-off (MWCO) membranes of 10 and 1 kDa resulted in the increased activity of each permeate, with a final IC50 value of 58.4 ± 4.6 µg mL⁻¹. Permeate containing peptides lower than 1 kDa was separated by reversed-phase high performance liquid chromatography (RP-HPLC). Four fractions (A-D) with potent ACE inhibitory activity were isolated and their main peptides identified using high performance liquid chromatography coupled to an electrospray ion trap Fourier transform ion cyclotron resonance-mass spectrometer (HPLC-ESI-IT-FTICR) followed by comparison with databases and de novo sequencing. The amino acid sequences of the identified peptides contained at least one hydrophobic and/or a proline together with positively charged residues in at least one of the three C-terminal positions. The IC50 values of the fractions ranged from 1.92 to 8.83 µg mL⁻¹, however this study fails to identify which of these peptides are ultimately responsible for the potent antihypertensive activity of these fractions.

Study design for the "effect of METOprolol in CARDioproteCtioN during an acute myocardial InfarCtion" (METOCARD-CNIC): a randomized, controlled parallel-group, observer-blinded clinical trial of early pre-reperfusion metoprolol administration in ST-segment elevation myocardial infarction.

BACKGROUND: Infarct size predicts post-infarction mortality. Oral ß-blockade within 24 hours of a ST-segment elevation acute myocardial infarction (STEMI) is a class-IA indication, however early intravenous (IV) ß-blockers initiation is not encouraged. In recent magnetic resonance imaging (MRI)-based experimental studies, the ß(1)-blocker metoprolol has been shown to reduce infarct size only when administered before coronary reperfusion. To date, there is not a single trial comparing the pre- vs. post-reperfusion ß-blocker initiation in STEMI. OBJECTIVE: The METOCARD-CNIC trial is testing whether the early initiation of IV metoprolol before primary percutaneous coronary intervention (pPCI) could reduce infarct size and improve outcomes when compared to oral post-pPCI metoprolol initiation. DESIGN: The METOCARD-CNIC trial is a randomized parallel-group single-blind (to outcome evaluators) clinical effectiveness trial conducted in 5 Counties across Spain that will enroll 220 participants. Eligible are 18- to 80-year-old patients with anterior STEMI revascularized by pPCI ≤6 hours from symptom onset. Exclusion criteria are Killip-class ≥III, atrioventricular block or active treatment with ß-blockers/bronchodilators. Primary end point is infarct size evaluated by MRI 5 to 7 days post-STEMI. Prespecified major secondary end points are salvage-index, left ventricular ejection fraction recovery (day 5-7 to 6 months), the composite of (death/malignant ventricular arrhythmias/reinfarction/admission due to heart failure), and myocardial perfusion. CONCLUSIONS: The METOCARD-CNIC trial is testing the hypothesis that the early initiation of IV metoprolol pre-reperfusion reduces infarct size in comparison to initiation of oral metoprolol post-reperfusion. Given the implications of infarct size reduction in STEMI, if positive, this trial might evidence that a refined use of an approved inexpensive drug can improve outcomes of patients with STEMI.

ApoB100/LDLR-/- hypercholesterolaemic mice as a model for mild cognitive impairment and neuronal damage.

Recent clinical findings support the notion that the progressive deterioration of cholesterol homeostasis is a central player in Alzheimer's disease (AD). Epidemiological studies suggest that high midlife plasma total cholesterol levels are associated with an increased risk of AD. This paper reports the plasma cholesterol concentrations, cognitive performance, locomotor activity and neuropathological signs in a murine model (transgenic mice expressing apoB100 but knockout for the LDL receptor [LDLR]) of human familial hypercholesterolaemia (FH). From birth, these animals have markedly elevated LDL-cholesterol and apolipoprotein B100 (apoB100) levels. These transgenic mice were confirmed to have higher plasma cholesterol concentrations than wild-type mice, an effect potentiated by aging. Further, 3-month-old transgenic mice showed cholesterol (total and fractions) concentrations considerably higher than those of 18-month-old wild-type mice. The hypercholesterolaemia of the transgenic mice was associated with a clear locomotor deficit (as determined by rotarod, grip strength and open field testing) and impairment of the episodic-like memory (determined by the integrated memory test). This decline in locomotor activity and cognitive status was associated with neuritic dystrophy and/or the disorganization of the neuronal microtubule network, plus an increase in astrogliosis and lipid peroxidation in the brain regions associated with AD, such as the motor and lateral entorhinal cortex, the amygdaloid basal nucleus, and the hippocampus. Aortic atherosclerotic lesions were positively correlated with age, although potentiated by the transgenic genotype, while cerebral ß-amyloidosis was positively correlated with genetic background rather than with age. These findings confirm hypercholesterolaemia as a key biomarker for monitoring mild cognitive impairment, and shows these transgenic mice can be used as a model for cognitive and psycho-motor decline.