I. Navarro-Blasco

Selenium, iodine, ω-3 PUFA and natural antioxidant from Melissa officinalis L.: A combination of components from healthier dry fermented sausages formulation

A new formulation of dry fermented sausage, including ingredients that improve the nutritional and health benefits of this type of product is presented. Se yeast (2g/kg), iodized salt (26g/kg), linseed:algae (3:2) emulsion (62.5g/kg), and lyophilized water extract of Melissa officinalis L. as a source of natural antioxidants (686mg/kg), yielded dry fermented sausages with technological and sensory properties similar to traditional ones. From the nutritional standpoint, a 50g portion of this product would give 100% of the recommended daily intake for Se, 70% of DRI for iodine, and 40% and 100% of the reference intake for alpha-linolenic and EPA+DHA, respectively. The omega-6/omega-3 ratio decreased from 15.7 in the control product to 1.96 in the modified one. Despite the high PUFA content, no oxidation was detected by TBARS (<0.15mgMDA/kg) and volatile compounds, showing the effectiveness of the natural antioxidants from M. officinalis. The modified formulation presented good acceptability for panelists with similar appearance, odor, taste and juiciness as the control products.

Solidification/stabilization of toxic metals in calcium aluminate cement matrices.

The ability of calcium aluminate cement (CAC) to encapsulate toxic metals (Pb, Zn and Cu) was assessed under two curing conditions. Changes in the consistency and in the setting time were found upon the addition of the nitrates of the target metals. Both Pb and Cu caused a delay in CAC hydration, while Zn accelerated the stiffening of the mortar. Compressive strengths of the metal-doped mortars, when initially cured at 60 °C/100% RH, were comparable with that of the free-metal mortar. Three different pore size distribution patterns were identified and related to the compounds identified by XRD and SEM. Sorbent capacities of CAC for the toxic metals were excellent: a total uptake was achieved for up to 3 wt.% loading of the three metals. In this way, CAC mortars were perfectly able to encapsulate the toxic metals, allowing the use of CAC for waste management as proved by the leaching tests.

Selenium content of Spanish infant formulae and human milk: Influence of protein matrix, interactions with other trace elements and estimation of dietary intake by infants

The selenium content of infant formulae varies as a result of differences in the amount of intrinsic selenium compounds. Manufacturers have been gradually changing the protein profile of infant formulae to reflect human milk contents more closely. Because of these variations in infant formula composition and their potential impact on selenium content, this trace element was analysed with regard to the different protein sources. The aims of this study were to determine the selenium content in infant formulae sold commercially in Spain, to estimate a daily dietary intake for infants fed on formulae and to compare with the selenium provided by Spanish breast milk samples used as a reference. We have also identified certain trace elements added to formulae which interact with selenium according to the type and protein matrix of the infant formulae. Selenium concentration was determined by inductively coupled plasma atomic emission spectrometry (ICP-AES) with a hydride generator. The selenium concentrations in human milk and infant formulae determined in this study are similar to those found by other researchers in different countries. The daily selenium intake from the formulae studied was estimated according to the recommended doses from the manufacturers. The theoretical selenium intake of nursed infants has been studied in relation to the Recommended Dietary Allowance (RDA: 10 microg Se/day) and the specific recommendations for infant formula nutrient contents (10-35 microg Se/L) set by the Expert Panel of life Sciences Research Office (LSRO) of the American Society for Nutritional Sciences. According to our results, on an overall view, infants fed on the studied infant formulae have an intake between basal and normative requirements. This might be considered as providing an adequate selenium supply. However, the intake of setenium provided by several formulae included in this research did not reach the RDA for the first month of neonate life.

High in omega-3 fatty acids bologna-type sausages stabilized with an aqueous-ethanol extract of Melissa officinalis.

A new formulation of bologna-type sausage enriched in ω-3 polyunsaturated fatty acids (PUFA) (8.75% linseed oil) was developed, using a lyophilized aqueous-ethanolic extract of Melissa officinalis. A comparison with the effectiveness of butylhydroxy anisole (BHA) synthetic antioxidant to decrease the oxidation of PUFAs was performed. The formulation increased the ω-3 PUFAs content, especially α-linolenic acid, decreasing significantly the ω-6/ω-3 ratio from 17.3 to 1.9, and also the Atherogenic Index and Thrombogenic Index (0.38-0.31 and 1.03-0.54, respectively). Modified sausages with BHA and Melissa extract showed significantly lower peroxides value (2.62 and 6.11 meq O₂/kg) and thiobarbituric acid value (0.26 and 0.27 mg malondialdehyde/kg) and higher antioxidant capacity (hydrophilic fraction ABTS: 0.45 and 0.74 meq Trolox/g product; lipofilic fraction ABTS: 0.44 and 0.37 meq Trolox/g product) than those without these ingredients (16.49 meq O₂/kg, 2.08 mg malondialdehyde /kg, 0.26 and 0.27 meq Trolox/g product, respectively). Sensorial tests showed that acceptability of the new formulations was similar to control products.

Optimization of a gelled emulsion intended to supply ω-3 fatty acids into meat products by means of response surface methodology

The optimization of a gelled oil-in-water emulsion was performed for use as fat replacer in the formulation of ω-3 PUFA-enriched cooked meat products. The linseed oil content, carrageenan concentration and surfactant-oil ratio were properly combined in a surface response design for maximizing the hardness and minimizing the syneresis of the PUFA delivery system. The optimal formulation resulted in a gelled emulsion containing 40% of oil and 1.5% of carrageenan, keeping a surfactant-oil ratio of 0.003. The gel was applied as a partial fat replacer in a Bologna-type sausage and compared to the use of an O/W emulsion also enriched in ω-3. Both experimental sausages contributed with higher ω-3 PUFA content than the control. No sensory differences were found among formulations. The selected optimized gelled oil-in-water emulsion was demonstrated to be a suitable lipophilic delivery system for ω-3 PUFA compounds and applicable in food formulations as fat replacer.

Long-term metabolic correction of Wilson’s disease in a murine model by gene therapy

BACKGROUND & AIMS Wilsons disease (WD) is an autosomal recessively inherited copper storage disorder due to mutations in the ATP7B gene that causes hepatic and neurologic symptoms. Current treatments are based on lifelong copper chelating drugs and zinc salts, which may cause side effects and do not restore normal copper metabolism. In this work we assessed the efficacy of gene therapy to treat this condition. METHODS We transduced the liver of the Atp7b(-/-) WD mouse model with an adeno-associated vector serotype 8 (AAV8) encoding the human ATP7B cDNA placed under the control of the liver-specific α1-antitrypsin promoter (AAV8-AAT-ATP7B). After vector administration we carried out periodic evaluation of parameters associated with copper metabolism and disease progression. The animals were sacrificed 6months after treatment to analyze copper storage and hepatic histology. RESULTS We observed a dose-dependent therapeutic effect of AAV8-AAT-ATP7B manifested by the reduction of serum transaminases and urinary copper excretion, normalization of serum holoceruloplasmin, and restoration of physiological biliary copper excretion in response to copper overload. The liver of treated animals showed normalization of copper content and absence of histological alterations. CONCLUSIONS Our data demonstrate that AAV8-AAT-ATP7B-mediated gene therapy provides long-term correction of copper metabolism in a clinically relevant animal model of WD providing support for future translational studies.

Aflatoxin levels and exposure assessment of Spanish infant cereals

Aflatoxins (AFB1, AFB2, AFG1 and AFG2) are immunosuppressant, mutagenic, teratogenic and carcinogenic agents with a widespread presence in foodstuffs. Since human exposure to aflatoxins occurs primarily by contaminated food intake, and given the greater susceptibility of infants to their adverse effects, the quantification of these mycotoxins in infant food based on cereals is of relevance. Aflatoxin levels were determined in 91 Spanish infant cereals classified in terms of non- and organically produced and several types from 10 different manufacturers, using a extraction procedure followed by inmunoaffinity column clean-up step and HPLC with fluorescence detection (FLD) and post-column derivatisation (Kobra Cell system). Daily aflatoxin intake was also assessed. Preliminary analysis showed a valuable incidence of detected infant cereal samples at an upper concentration level than the detection limit for total aflatoxin (66%), corresponding to a 46, 40, 34 and 11% for AFB1, AFB2, AFG1 and AFG2, respectively. Lower aflatoxin values (median, Q1, Q3) in conventional infant cereal (n = 74, AFB1: <LOD (n.d.; 0.02), AFB2: n.d. (n.d.; 0.01), AFG1: <LOD (n.d.; 0.004), and AFG2: n.d. (n.d.; <LOD) and total AF (AFtotal): 0.01 (<LOD; 0.04  µg kg−1) in comparison with infant cereal ecologically produced (n = 17, AFB1: 0.02 (0.02; 0.21), AFB2: n.d. (n.d.; 0.03), AFG1: 0.02 (0.01; 0.05), and AFG2: 0.007 (n.d.; 0.02) and AFtotal: 0.05 (0.03; 0.31  µg kg−1) were found. In addition, five organic formulations (3.11, 1.98, 0.94, 0.47 and 0.21  µg kg−1) exceeded European AFB1 legislation (0.10 µg kg−1) versus two conventional cereals (0.35 and 0.12  µg kg−1). According to the type of infant cereal, those with cocoa had the highest aflatoxin levels. Gluten‐free and cereals with dehydrated fruits had an intermediate level and milk- or honey-based cereals and multi-cereals contained the lowest levels. With the exception of the non-compliant cocoa-based organic formulation, none of the infant cereals analyzed gave a higher intake of 1 ng kg−1 body weight per day, suggesting that infants fed on infant cereals are exposed to a low health hazard. Nevertheless, manufacturers are advised for continued efforts in routine monitoring and a more careful selection of raw material to minimize aflatoxin levels in these infant foods.

Healthy reduced-fat bologna sausages enriched in ALA and DHA and stabilized with Melissa officinalis extract

Reduced-energy and reduced-fat Bologna products enriched with ω-3 polyunsaturated fatty acids were formulated by replacing the pork back-fat by an oil-in-water emulsion containing a mixture of linseed-algae oil stabilized with a lyophilized Melissa officinalis extract. Healthier composition and lipid profile was obtained: 85 kcal/100 g, 3.6% fat, 0.6 g ALA and 0.44 g DHA per 100 g of product and ω-6/ω-3 ratio of 0.4. Technological and sensory problems were not detected in the new formulations. Reformulation did not cause oxidation problems during 32 days of storage under refrigeration. The results suggest that it is possible to obtain reduced-fat Bologna-type sausages rich in ALA and DHA and stabilized with natural antioxidants, applying the appropriate technology without significant effects on the sensory quality, yielding interesting products from a nutritional point of view.

Encapsulation, solid-phases identification and leaching of toxic metals in cement systems modified by natural biodegradable polymers.

Cement mortars loaded with Cr, Pb and Zn were modified by polymeric admixtures [chitosans with low (LMWCH), medium (MMWCH) and high (HMWCH) molecular weight and hydroxypropylchitosan (HPCH)]. The influence of the simultaneous presence of the heavy metal and the polymeric additive on the fresh properties (consistency, water retention and setting time) and on the compressive strength of the mortars was assessed. Leaching patterns as well as properties of the cement mortars were related to the heavy metals-bearing solid phases. Chitosan admixtures lessened the effect of the addition of Cr and Pb on the setting time. In all instances, chitosans improved the compressive strength of the Zn-bearing mortars yielding values as high as 15 N mm(-2). A newly reported Zn phase, dietrichite (ZnAl(2)(SO(4))(4)·22H(2)O) was identified under the presence of LMWCH: it was responsible for an improvement by 24% in Zn retention. Lead-bearing silicates, such as plumalsite (Pb(4)Al(2)(SiO(3))(7)), were also identified by XRD confirming that Pb was mainly retained as a part of the silicate network after Ca ion exchange. Also, the presence of polymer induced the appearance and stabilization of some Pb(IV) species. Finally, diverse chromate species were identified and related to the larger leaching values of Cr(VI).

Aluminium content of Spanish infant formula

Levels of aluminium in 82 different infant formulae from nine different manufacturers in Spain were determined by acid-microwave digestion and graphite furnace atomic absorption spectrophotometry. The influence of aluminium content in tap water in reconstituted powder formulae was examined and an estimate was made of the theoretical toxic aluminium intake in comparison with the provisional tolerable weekly intake (PTWI). Possible interactions between aluminium and certain essential trace elements added to infant formulations have been studied according to the type or main protein-based infant formula. In general, the infant formulae contained a higher aluminium content than that found in human milk, especially in the case of soya, preterm or hydrolysed casein-based formulae. Standard formulae gave lower aluminium intakes amounting to about 4% PTWI. Specialized and preterm formulae resulted in a moderate intake (11–12 and 8–10% PTWI, respectively) and soya formulae contributed the highest intake (15% PTWI). Aluminium exposure from drinking water used for powder formula reconstitution was not considered a potential risk. In accordance with the present state of knowledge about aluminium toxicity, it seems prudent to call for continued efforts to standardize routine quality control and reduce aluminium levels in infant formula as well as to keep the aluminium concentration under 300 μg l-1 for all infant formulae, most specifically those formulae for premature and low birth neonates.