Simultaneous microwave-assisted extraction of bioactive compounds from aged garlic.

In this work, the simultaneous extraction of bioactives (organosulfur compounds, such as S-allyl-L-cysteine (SAC), carbohydrates, such as neokestose and neonystose, and total phenolic compounds) from aged garlic has been optimized for the first time to obtain multifunctional extracts for further application as food ingredients. Analytical methods using liquid chromatography coupled to mass spectrometry (HPLC-MS) and by hydrophilic interaction liquid chromatography with evaporative light scattering detection (HILIC-ELSD) were also previously optimized. High sensitivity (limits of detection between 0.013 and 0.77 µg mL-1) and appropriate repeatability (< 12%) and accuracy (> 92%) for the analysis of bioactives were achieved. After selecting water as the extraction solvent and microwave-assisted extraction (MAE) as the most efficient technique, operation conditions were optimized using a Box-Behnken experimental design (60 min; 120 °C; 0.05 g mL-1; 1 cycle) to maximize the content of bioactives from different aged garlic samples. Regarding organosulfur compounds, only SAC (traces-2.32 mg g-1 dry sample) and cycloalliin (1.23-3.01 mg g-1 dry sample) were detected in all samples, while amino acids such as arginine (0.24-3.45 mg g-1 dry sample) and proline (0.43-3.91 mg g-1 dry sample) were, in general, the most abundant. Bioactive carbohydrates (from trisaccharides to nonasaccharides) were only detected in fresh garlic and aged garlic processed under mild conditions, whereas all garlic extracts showed antioxidant activity. The developed MAE methodology is shown as a successful alternative to other procedures for the simultaneous extraction of aged garlic bioactives intended by the food and nutraceutical industries, among others.

Exploitation of artichoke byproducts to obtain bioactive extracts enriched in inositols and caffeoylquinic acids by Microwave Assisted Extraction.

Byproducts from artichoke represent the majority of the mass collected from the plant and constitute an interesting source of bioactive compounds such as inositols and caffeoylquinic acids. In this work, a microwave assisted extraction (MAE) methodology was developed for the simultaneous extraction of these compounds from artichoke stalks, leaves, receptacles and external bracts. Optimal MAE conditions to maximize the extraction of these bioactives and the antioxidant activity were 97 °C, 3 min, ethanol:water (50:50, v/v). Moreover, a GC-MS methodology was also developed for the simultaneous determination of these compounds in a single run; optimal derivatization conditions were achieved using hexamethyldisilazane and N,O-bis(trimethylsilyl)trifluoroacetamide with 1% trimethylchlorosilane. Artichoke receptacle extracts were the richest in caffeoylquinic acids (28-35 mg g-1 dry sample), followed by the bracts (9-18 mg g-1 dry sample), while those from leaves showed the highest concentrations of inositols (up to 15 mg g-1 dry sample). Receptacle extracts also had the highest antioxidant activity (123 mg TE g-1 dry sample) and the greatest concentration of total phenolic compounds (47 mg GAE g-1 dry sample). Therefore, the developed methodology could be considered as a valuable procedure to obtain and characterize bioactive ingredients with industrial interest from artichoke byproducts, opening new routes of revalorization of artichoke agro-industrial residues.

Structural changes of a protein extract from apple with polyphenoloxidase activity obtained by cationic reversed micellar extraction induced by high-pressure carbon dioxide and thermosonication.

Polyphenoloxidase from apple was extracted and further concentrated by cationic reversed micellar extraction. Previous to reversed micellar extraction a crude protein extract was obtained using AG2-X8 as adsorbent of phenolic compounds and the detergent Triton X-100. Forward and backward extraction conditions were optimized by using dodecyl trimethyl ammonium bromide as surfactant in the organic phase. Optimization was carried out to obtain the highest value of PPO activity recovery and the purification fold at the different experimental conditions. Under the optimum extraction conditions, PPO activity recovery was 99% and purification fold reached a value of 17, showing that reversed micellar extraction was a good technique as a first step to concentrate on a targeted enzyme. After removing some impurities by centrifuge ultrafiltration, the protein extract with PPO activity was treated by pressurized carbon dioxide and thermosonication achieving residual PPO activity values of 16 ± 3 and 9 ± 1%, respectively. Quenching experiments by iodide performed in the non-treated extract and in the treated extracts revealed conformational changes of this protein fraction reflected in the greater exposure of the fluorophore to the quencher.

Gas chromatographic-based techniques for the characterization of low molecular weight carbohydrates and phenylalkanoid glycosides of Sedum roseum root supplements.

An extensive characterization of low molecular weight carbohydrates (LMWC) and phenylalkanoid glycosides (PAG) of Sedum roseum root supplements has been carried out for the first time by gas chromatography coupled to mass spectrometry (GC-MS) and by comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC × GC-ToF MS). Optimization of the required derivatization procedure for improved determination of PAG showed the combined use of trimethylsilylimidazole and trimethylchlorosilane as the most appropriate reagents. Up to 37 compounds were qualitative- and quantitatively characterized in different dietary supplements of S. roseum by GC-MS. In addition to the well-known rosin, rosarin, rosavin and salidroside, other carbohydrates, polyalcohols, acids, etc. were determined. Among them, several seven-carbon monosaccharides such as coriose and 2,7-anhydro-ß-d-altro-heptulose were detected for the first time in S. roseum root supplements. Sedoheptulose was found to be the most abundant compound (9-151 mg g-1), followed by rosiridin (20-81 mg g-1) and rosavin (11-56 mg g-1). The use of GC × GC-ToF MS allowed the detection and tentative assignation of 48 additional compounds mainly belonging to the phenylalkanoid glycoside, pentosyl-hexose and hexosyl-hexose families.

Predictive factors of hospital stay, mortality and functional recovery after surgery for hip fracture in elderly patients. / Factores determinantes de estancia hospitalaria, mortalidad y evolución funcional tras cirugía por fractura de cadera en el anciano.

OBJECTIVES: Due to its high prevalence and serious consequences it is very important to be well aware of factors that might be related to medical complications, mortality, hospital stay and functional recovery in elderly patients with hip fracture. MATERIAL AND METHODS: A prospective study of a group of 130 patients aged over 75 years admitted for osteoporotic hip fracture. Their medical records, physical and cognitive status prior to the fall, fracture type and surgical treatment, medical complications and functional and social evolution after hospitalization were evaluated. RESULTS: Patients with greater physical disability, more severe cognitive impairment and those who lived in a nursing home before the fracture had worse functional recovery after surgery. Treatment with intravenous iron to reduce transfusions reduced hospital stay and improved walking ability. Infections and heart failure were the most frequent medical complications and were related to a longer hospital stay. The prescription of nutritional supplements for the patients with real indication improved their physical recovery after the hip fracture CONCLUSIONS: Evaluation of physical, cognitive and social status prior to hip fracture should be the basis of an individual treatment plan because of its great prognostic value. Multidisciplinary teams with continuous monitoring of medical problems should prevent and treat complications as soon as possible. Intravenous iron and specific nutritional supplements can improve functional recovery six months after hip fracture.

Estrategias para la implantación del cribado poblacional de cáncer de cuello uterino con test VPH-AR / Strategies for the implementation of population-based screening for cervical cancer with the HR-HPV test

Introducción: desde el año 2005 se trabaja coordinadamente Atención Primaria y especializada para revertir el cribado de cáncer de cuello de útero de oportunista a poblacional. En 2011 se añadió el test de virus de papiloma humano de alto riesgo a la citología (co-test) como cribado primario. Objetivo: potenciar el cribado poblacional de cáncer de cuello de útero. Material y métodos: captación activa de la población a través de las matronas de Atención Primaria y atención en ginecología de las mujeres positivas a virus de papiloma humano de alto riesgo. En Anatomía Patológica se realiza control de calidad de las pruebas y se monitorizan los resultados. Resultados: se ha duplicado la cobertura poblacional, el 80,7% de las pruebas de cribado se realiza en Atención Primaria y el número de neoplasias intraepiteliales cervicales se ha quintuplicado. Conclusiones: la implicación de todos los profesionales en el cribado ha permitido conseguir una cobertura del 66,6% y la introducción del test de virus de papiloma humano de alto riesgo ha incrementado los diagnósticos de neoplasias intraepiteliales cervicales (AU)

Introduction: Since 2005, a coordinated effort by primary care and gynaecology services has shifted screening from an opportunistic setting to a population-based strategy. High-risk Human papilloma viruses testing was added to the Papanicolau smear (co-testing) as the primary screening test in 2011. Objective: To implement population-based cancer screening. Material and methods: Primary care midwives arrange appointments for women and, if tested positive, they are then attended in gynaecology services. Quality control and the monitoring of results is carried out by the pathology service. Results: Coverage has doubled, 80% of screening tests are performed in primary care and the number of CIN 2/3 intraepithelial neoplasia cases have increased 5 fold. Conclusion: The participation of the entire health screening team has increased coverage to 66%. The introduction of the human papilloma viruses test has increased the detection of intraepithelial neoplasia cases (AU)

Experimental study showed that adding fortifier and extra-hydrolysed proteins to preterm infant mothers' milk increased osmolality.

AIM: Measuring milk osmolality after adjustable fortification is clinically relevant, as values exceeding recommended safety thresholds might result in gastrointestinal consequences. The aim of this study was to evaluate the effect of four fortification levels and storage time on the osmolality of human milk. METHODS: This was an experimental study using 71 spare samples of fresh breastmilk collected from 31 mothers of preterm infants. Osmolality was measured before and after adding commercial human milk fortifier containing dextrinomaltose and hydrolysed proteins at four different concentrations. Measurements were performed at various points during the 23 hours after fortification. RESULTS: The mean basal osmolality of the 71 human milk samples was 296 ± 14 milliosmoles (mOsm)/kg, and these remained stable over a period of 23 hours. Just after fortification, the four fortified formulas showed higher osmolalities than the nonfortified human milk, ranging between 384 ± 14 and 486 ± 15 mOsm/kg, respectively (p < 0.01). This osmolality increased significantly from 20 minutes to 23 hours after fortification (p < 0.05). CONCLUSION: Adding fortifier and extra-hydrolysed proteins to human preterm milk increased osmolality, and these osmolality levels also increased with time. We recommend evaluating the risk of hyperosmolality when a higher fortification level is needed, to avoid gastrointestinal problems.

Is the ISAC 112 Microarray Useful in the Diagnosis of Pollinosis in Spain?

BACKGROUND: Multiple sensitization is frequent among pollen-allergic patients. The goal of this study was to determine the diagnostic accuracy of the ImmunoCAP ISAC 112 (ISAC112) microarray in allergy to pollen from several taxa and its clinical utility in a Spanish population. METHODS: Specific IgE was determined in 390 pollen-allergic patients using the ISAC112 microarray. Diagnostic accuracy (sensitivity, specificity, predictive values, and area under the ROC curve) was calculated for the diagnosis of allergy to pollen from grass (n=49), cypress (n=75), olive tree (n=33), plane tree (n=63), and pellitory of the wall (n=17) and compared with that of the singleplex ImmunoCAP immunoassay. RESULTS: The sensitivity of the ISAC112 microarray ranged from 68.2% for allergy to plane tree pollen to 93.9% for allergy to grass pollen. The specificity was >90%. The AUC for the diagnosis of allergy to plane tree pollen was 0.798, whereas the AUC for the remaining cases was ≥0.876. The accuracy of ISAC112 was higher than that of ImmunoCAP for plane tree pollen and similar for the remaining pollens. The frequency of sensitization to most species-specific allergenic components and profilins varied between the different geographical regions studied. A total of 73% of pollen-allergic patients were sensitized to species-specific components of more than 1 pollen type. CONCLUSIONS: The ISAC112 microarray is an accurate tool for the diagnosis of allergy to pollen from grass, cypress, olive tree, plane tree, and pellitory of the wall. The features of the ISAC112 microarray are similar or superior (in the case of plane tree pollen) to those of ImmunoCAP. This microarray is particularly useful for the etiologic diagnosis of pollinosis in patients sensitized to multiple pollen species whose pollination periods overlap.

Pressurized liquid extraction of Aglaonema sp. iminosugars: Chemical composition, bioactivity, cell viability and thermal stability.

Pressurized liquid extraction of Aglaonema sp. iminosugars has been optimized. A single cycle under optimal conditions (80mg, 100°C, 2min) was enough to extract ⩾96% of most iminosugars. Further incubation with Saccharomyces cerevisiae for 5h removed coextracted interfering low molecular weight carbohydrates from extracts of different Aglaonema cultivars. A complete characterization of these extracts was carried out by gas chromatography-mass spectrometry: three iminosugars were tentatively identified for the first time; α-homonojirimycin and 2,5-dideoxy-2,5-imino-d-mannitol were the major iminosugars determined. α-Glucosidase inhibition activity, cell viability and thermal stability of Aglaonema extracts were also evaluated. Extracts with IC50 for α-glucosidase activity in the 0.010-0.079mgmL(-1) range showed no decrease of Caco-2 cell viability at concentrations lower than 125µgmL(-1) and were stable at 50°C for 30days. These results highlight the potential of Aglaonema extracts as a source of bioactives to be used as functional ingredients.

Characterization of goat colostrum oligosaccharides by nano-liquid chromatography on chip quadrupole time-of-flight mass spectrometry and hydrophilic interaction liquid chromatography-quadrupole mass spectrometry.

A detailed qualitative and quantitative characterization of goat colostrum oligosaccharides (GCO) has been carried out for the first time. Defatted and deproteinized colostrum samples, previously treated by size exclusion chromatography (SEC) to remove lactose, were analyzed by nanoflow liquid chromatography-quadrupole-time of flight mass spectrometry (Nano-LC-Chip-Q-TOF MS). Up to 78 oligosaccharides containing hexose, hexosamine, fucose, N-acetylneuraminic acid or N-glycolylneuraminic acid monomeric units were identified in the samples, some of them detected for the first time in goat colostra. As a second step, a hydrophilic interaction liquid chromatography coupled to mass spectrometry (HILIC-MS) methodology was developed for the separation and quantitation of the main GCO, both acidic and neutral carbohydrates. Among other experimental chromatographic conditions, mobile phase additives and column temperature were evaluated in terms of retention time, resolution, peak width and symmetry of target carbohydrates. Narrow peaks (wh: 0.2-0.6min) and good symmetry (As: 0.8-1.4) were obtained for GCO using an acetonitrile:water gradient with 0.1% ammonium hydroxide at 40°C. These conditions were selected to quantify the main oligosaccharides in goat colostrum samples. Values ranging from 140 to 315mgL(-1) for neutral oligosaccharides and from 83 to 251mgL(-1) for acidic oligosaccharides were found. The combination of both techniques resulted to be useful to achieve a comprehensive characterization of GCO.

Analysis of iminosugars and other low molecular weight carbohydrates in Aglaonema sp. extracts by hydrophilic interaction liquid chromatography coupled to mass spectrometry.

A method by hydrophilic interaction liquid chromatography coupled to tandem mass spectrometry (HILIC-MS(2)) has been successfully developed for the simultaneous analysis of bioactive iminosugars and other low molecular weight carbohydrates in Aglaonema leaf extracts. Among other experimental chromatographic conditions, mobile phase eluents, additives and column temperature were evaluated in terms of retention time, resolution, peak width and symmetry provided for target carbohydrates. In general, narrow peaks (wh: 0.2-0.6min) with good symmetry (As: 0.9-1.3) and excellent resolution (Rs>1.8) were obtained for iminosugars using an acetonitrile:water gradient with 5mM ammonium acetate in both eluents at 55°C. Tandem mass spectra were used to confirm the presence of previously detected iminosugars in Aglaonema extracts and to tentatively identify for the first time others such as miglitol isomer, glycosyl-miglitol isomers and glycosyl-DMDP isomers. Concentration of total iminosugars varied from 1.35 to 2.84mgg(-1) in the extracts of the different Aglaonema samples analyzed. To the best of our knowledge, this is the first time that a HILIC-MS(2) method has been proposed for the simultaneous analysis of iminosugars and other low molecular weight carbohydrates of Aglaonema sp. extracts.

CYP11A1 expression in bone is associated with aromatase inhibitor-related bone loss.

Aromatase inhibitors (AIs) used as adjuvant therapy in postmenopausal women with hormone receptor-positive breast cancer cause diverse musculoskeletal side effects that include bone loss and its associated fracture. About half of the 391 patients treated with AIs in the Barcelona-Aromatase induced bone loss in early breast cancer cohort suffered a significant bone loss at lumbar spine (LS) and/or femoral neck (FN) after 2 years on AI-treatment. In contrast, up to one-third (19.6% LS, 38.6% FN) showed no decline or even increased bone density. The present study aimed to determine the genetic basis for this variability. SNPs in candidate genes involved in vitamin D and estrogen hormone-response pathways (CYP11A1, CYP17A1, HSD3B2, HSD17B3, CYP19A1, CYP2C19, CYP2C9, ESR1, DHCR7, GC, CYP2R1, CYP27B1, VDR and CYP24A1) were genotyped for association analysis with AI-related bone loss (AIBL). After multiple testing correction, 3 tag-SNPs (rs4077581, s11632698 and rs900798) located in the CYP11A1 gene were significantly associated (P<0.005) with FN AIBL at 2 years of treatment. Next, CYP11A1 expression in human fresh bone tissue and primary osteoblasts was demonstrated by RT-PCR. Both common isoforms of human cholesterol side-chain cleavage enzyme (encoded by CYP11A1 gene) were detected in osteoblasts by western blot. In conclusion, the genetic association of CYP11A1 gene with AIBL and its expression in bone tissue reveals a potential local function of this enzyme in bone metabolism regulation, offering a new vision of the steroidogenic ability of this tissue and new understanding of AI-induced bone loss.

¿Impacto de la concentración mínima inhibitoria de vancomicina o incorrecto manejo terapéutico en el fracaso clínico por Staphylococcus aureus resistentes a meticilina? / Impact of vancomycin minimum inhibitory concentration or inadequate therapeutic management in the clinical outcome failure due to methicillin-resistant Staphylococcus aureus?

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Are basophil activation and sulphidoleukotriene determination useful tests for monitoring patients with peach allergy receiving sublingual immunotherapy with a Pru p 3-enriched peach extract?

INTRODUCTION: Treatment of food allergy essentially consists of food avoidance, but immunotherapy with food is emerging as a new therapeutic option. OBJECTIVE: To evaluate clinical improvement and immunological changes in patients with peach allergy following sublingual immunotherapy (SLIT) with a Prup3 quantified peach extract. METHODS: A randomized, double-blind, placebo-controlled clinical trial with peach SLIT was conducted. We assessed clinical efficacy after 6 months of treatment by means of double-blind, placebo-controlled oral challenges with peach and also evaluated immunological changes (basophil activation test [BAT] and determination of sulphidoleukotriene production) following stimulation with peach peel and pulp, rPrup3, rMald 1, and rMal d 4 stimulation. We also measured specific IgE and IgG4 to Pru p3. RESULTS: After 6 months of SLIT (T6), the active group showed a 3-fold improvement in tolerance to Prup3 and a significant increase in IgE to rPrup3 and in sLT production following stimulation with peach peel and rPrup3. There was also a significant increase in BAT results after stimulation with rPrup3 at 1 month of SLIT (T1). Statistically significant between-group differences were only observed for BAT with peach peel and pulp at T1 and T6 and for BAT with rPru p3 at T6. No changes were observed in BAT with rMal d 1 or rMal d 4 or in IgG4 levels to nPrup3. CONCLUSIONS: SLIT with a Pru p 3 quantified peach extract is clinically effective and leads to an increase in basophil activation and sulphidoleukotriene production following stimulation with rPru p3 and peach peel in the first months of treatment.

Efficacy of a pectin-lecithin complex for treatment and prevention of gastric ulcers in horses.

The objective of this study was to evaluate the effect of a commercial feed supplement containing pectin-lecithin on squamous mucosa ulceration in horses exposed to an experimental ulceration model. Five mares were treated while five mares were controls for this crossover, blinded study. The mares were fed concentrates and hay and were stabled with a two-hour turn out per day for a period of four weeks. The pectin-lecithin complex was fed for the duration of the study on the treated group. At the end of a four-week period, all mares underwent a seven-day alternating feed deprivation (week 5). The study was repeated again after a four-week washout period. Gastroscopy was performed on days 1, 28 and 35 of the study and was digitally recorded. Independent evaluation of the recordings and scoring of the lesions using the Equine Gastric Ulcer Syndrome (EGUS), severity and number scores were performed by three experienced gastroscopists. The prevalence and severity of squamous ulcers significantly increased after intermittent feed deprivation (P<0.001). No significant effect of the treatment was observed (P>0.05). In this study, the addition of a commercially available pectin-lecithin complex to the feed of horses for five weeks did not prevent or minimise the risk for gastric ulceration of the squamous mucosa.

Homeopatía y productos dietéticos que no precisan prescripción médica en niños: ¿son siempre inofensivos? / Homeopathy and over-the-counter diet products in children: Are they always inoffensive?

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Developing implant dentistry education in Europe: the continuum from undergraduate to postgraduate education and continuing professional development.

INTRODUCTION: Implant dentistry is a treatment modality which has mainstream clinical practice of comprehensive care, which however is not adequately represented in the undergraduate dental curricula. A consensus workshop organised by ADEE in 2008, set the benchmarks for the knowledge and competences a modern dental practitioner must possess with regard to implant dentistry, as well as defined undergraduate and postgraduate pathways for the acquisition of these competences. Today, 5 years later, there exist several challenges for the implementation of these benchmarks in both undergraduate curricula but also post-graduation educational pathways. METHODS: A consensus workshop was organised by ADEE, bringing together 48 opinion leaders, including academic teachers of all disciplines related to implant dentistry, specialists, representatives of relevant scientific and professional associations, as well as industry delegates. The objectives of the workshop were to evaluate the existing scientific literature, reported experience and best practices in order to identify potential and limitations for the implementation of implant dentistry in the undergraduate curriculum, as well produce recommendations for the optimal educational structures for postgraduate programmes and continuing professional development. RESULTS: The scientific committee conducted two European-wide questionnaire surveys to better document the current state of education in implant dentistry. Upon completion of the surveys, reviewers were appointed to produce three scientific review papers, identifying current achievements and future challenges. Finally, during the 3 days of the workshop, all the evidence was reviewed and the main conclusions and recommendations that were adopted by all participants are reported in the present Consensus Paper. CONCLUSIONS: Implementation of implant dentistry in the undergraduate curriculum has improved significantly, but still lags behind the benchmarks set in 2008 and the diversity between institutions remains big. At the post-graduation level, there is currently a wide diversity of courses and pathways towards competences related to implant dentistry and there is at present a great need for quality assurance, as well as standardisation and transparency of the learning outcomes.

Optimization of pressurized liquid extraction of inositols from pine nuts (Pinus pinea L.).

Pressurized liquid extraction (PLE) has been used for the first time to extract bioactive inositols from pine nuts. The influence of extraction time, temperature and cycles of extraction in the yield and composition of the extract was studied. A quadratic lineal model using multiple linear regression in the stepwise mode was used to evaluate possible trends in the process. Under optimised PLE conditions (50°C, 18 min, 3 cycles of 1.5 mL water each one) at 10 MPa, a noticeable reduction in extraction time and solvent volume, compared with solid-liquid extraction (SLE; room temperature, 2h, 2 cycles of 5 mL water each one) was achieved; 5.7 mg/g inositols were extracted by PLE, whereas yields of only 3.7 mg/g were obtained by SLE. Subsequent incubation of PLE extracts with Saccharomyces cerevisiae (37°C, 5h) allowed the removal of other co-extracted low molecular weight carbohydrates which may interfere in the bioactivity of inositols.