Use of urea-polyacrylamide electrophoresis for discrimination of A1 and A2 beta casein variants in raw cow's milk.

Beta-casein (BC) in cow's milk occurs in several genetic variants, where BC A1 (BCA1) and BC A2 (BCA2) are the most frequent. This work deals with a method based on modified polyacrylamide gel electrophoresis using urea PAGE to discriminate BCA1 and BCA2 variants from Holstein Friesian (HF) and genetically selected Jersey A2/A2 (JA2) cow's milk. Two well defined bands were obtained from BC fraction of HF milk, while that of JA2 showed a single band. Proteins from these bands were sequenced by HPLC-quadrupole linear ion trap/mass spectrometry, resulting in BCA1 and BCA2 separation from the BC fraction of HF milk, whereas BCA2 was the only constituent of JA2 fraction. This method represents a feasible and useful tool to on site phenotyping of BC fraction of cow's milk for pharmaceutical and food industries applications.

Effect of starch-beeswax coatings on quality parameters of blackberries (Rubus spp.).

There is increased interest in berry fruits due to health benefits, and maintenance of fruit quality for longer periods of time has been a priority. We previously found that starch based coatings applied on raspberries was associated to volatile compounds production due to anoxic conditions. The objective of this work was to design more hydrophobic coatings with reduced thickness. A starch-beeswax dispersion containing 2 % (w/v) modified tapioca starch added with either 0.5 or 1.0 % (w/v) beeswax microparticles was produced, and used for spray coating freshly harvested blackberries (Rubus spp.). Coatings were air dried, packed in plastic trays and stored up to 16 days at 4 °C and 88 % relative humidity. Storage quality parameters such as hardness, respiration rate, anthocyanins content, total phenols, color changes and weight loss were evaluated. We did not find Interactions among coating ingredients, and incorporation of beeswax reduced moisture transfer rate. Coatings did not occlude the stomata and apparently did not over-hydrate the cuticle. This characteristic allowed appropriate gas exchange (O2 and CO2), and reduced accumulation of volatile compounds associated to fermentative metabolism. Respiration rates were 4.207 ± 0.157, 4.557 ± 0.220 and 4.780 ± 0.050 mmol CO2 kg(-1) h(-1) for control, 0.5 and 1 % of wax content in coatings, respectively. However, ethylene production increased throughout storage time along with beeswax concentration, indicating stressful conditions for the fruit. This trend appears to be related with changes in total phenols and anthocyanins during storage. Edible coatings based on starch and hydrophobic particles should be reformulated to maintain quality of stored berry fruits.

Antimicrobial edible films and coatings for meat and meat products preservation.

Animal origin foods are widely distributed and consumed around the world due to their high nutrients availability but may also provide a suitable environment for growth of pathogenic and spoilage microorganisms. Nowadays consumers demand high quality food with an extended shelf life without chemical additives. Edible films and coatings (EFC) added with natural antimicrobials are a promising preservation technology for raw and processed meats because they provide good barrier against spoilage and pathogenic microorganisms. This review gathers updated research reported over the last ten years related to antimicrobial EFC applied to meat and meat products. In addition, the films gas barrier properties contribute to extended shelf life because physicochemical changes, such as color, texture, and moisture, may be significantly minimized. The effectiveness showed by different types of antimicrobial EFC depends on meat source, polymer used, film barrier properties, target microorganism, antimicrobial substance properties, and storage conditions. The perspective of this technology includes tailoring of coating procedures to meet industry requirements and shelf life increase of meat and meat products to ensure quality and safety without changes in sensory characteristics.

The combined effect of diffuse radiation and leaf wetness on functional traits and transpiration efficiency on a cloud forest species.

Cloud forests are unique biomes that thrive in foggy environments for a substantial part of the season. Fog in cloud forests plays two critical roles: it reduces incoming radiation and creates a humid environment, leading to the wetting of the canopy. This paper aims to investigate the combined effect of both radiation and wetness on Myrica faya Wilbur-a cloud forest species present in subtropical regions-both directly in plants and through simulations. Experiments consisted of a controlled environment with two levels of radiation and leaf wetness: low radiation/wet conditions, and high radiation/no-wetness; and three treatments: continuous low radiation and wetness, continuous high radiation and no wetness and alternate high low radiation and alternate wetness. The results revealed that a combination of low radiation and leaf wetness significantly improves leaf stomata conductance and increases the specific leaf area (SLA). Changes in SLA were driven by leaf size changes. However, the minimum leaf conductance (gmin) did not respond to any of the treatments. The simulations focused on exploring the impact of radiation and canopy wetness on transpiration efficiency (TE), i.e. the ratio between photosynthesis (An) and transpiration (Tc). The simulations demonstrated that TE increased exponentially as the canopy was gradually wetted, regardless of the radiation environment. This increase in TE results from Tc approaching zero while An maintains positive values. Overall, this study provides an integrated understanding of how fog alters M. faya functioning and, potentially, other cloud forest tree species.

Electrostatic Fermentation: Molecular Response Insights for Tailored Beer Production.

Electrostatic fermentation avoids the cellular redox imbalance of traditional fermentation, but knowledge gaps exist. This study explores the impact of electrostatic fermentation on the growth, volatile profile, and genetic response of Saccharomyces pastorianus Saflager S-23. The applied voltage (15 and 30 V) in the electrostatic fermentation system increased the growth and substrate utilization of S. pastorianus while decreasing ethanol production. The aromas typically associated with traditional fermentation, such as alcoholic, grape, apple, and sweet notes, were diminished, while aromas like roses, fruits, flowers, and bananas were augmented in electrostatic fermentation. RNA-seq analysis revealed upregulation of genes involved in cell wall structure, oxidoreductase activity, and iron ion binding, while genes associated with protein synthesis, growth control, homeostasis, and membrane function were downregulated under the influence of applied voltage. The electrostatic fermentation system modulates genetic responses and metabolic pathways in yeast, rendering it a promising method for tailored beer production. Demonstrating feasibility under industrial-scale and realistic conditions is crucial for advancing towards commercialization.

A Liposomal Formulation to Exploit the Bioactive Potential of an Extract from Graciano Grape Pomace.

Antioxidant compounds with health benefits can be found in food processing residues, such as grape pomace. In this study, antioxidants were identified and quantified in an extract obtained from Graciano red grape pomace via a green process. The antioxidant activity of the extract was assessed by the DPPH and FRAP tests, and the phenolic content by the Folin-Ciocalteu test. Furthermore, nanotechnologies were employed to produce a safe and effective formulation that would exploit the antioxidant potential of the extract for skin applications. Anthocyanins, flavan-3-ols and flavanols were the main constituents of the grape pomace extract. Phospholipid vesicles, namely liposomes, were prepared and characterized. Cryo-TEM images showed that the extract-loaded liposomes were predominantly spherical/elongated, small, unilamellar vesicles. Light scattering results revealed that the liposomes were small (~100 nm), homogeneously dispersed, and stable during storage. The non-toxicity of the liposomal formulation was demonstrated in vitro in skin cells, suggesting its possible safe use. These findings indicate that an extract with antioxidant properties can be obtained from food processing residues, and a liposomal formulation can be developed to exploit its bioactive value, resulting in a promising healthy product.

Untargeted Metabolomic Liquid Chromatography High-Resolution Mass Spectrometry Fingerprinting of Apple Cultivars for the Identification of Biomarkers Related to Resistance to Rosy Apple Aphid.

Liquid chromatography high-resolution mass spectrometry fingerprinting together with pattern recognition techniques was used to determine the metabolites involved in the susceptibility of apple cultivars to rosy apple aphid (RAA). Preprocessing of ultra-high-performance liquid chromatography coupled to electrospray ionization and quadrupole time-of-flight mass spectrometry raw data of resistant and susceptible apple cultivars was carried out with XCMS and CAMERA packages. Univariate statistical tools and multivariate data analysis highlighted significant different profiles of the apple metabolomes according to their tolerance to RAA. Optimized and cross-validated Partial least squares discriminant analysis and orthogonal projections to latent structures discriminant analysis models confirmed trans-4-caffeoylquinic acid and 4-p-coumaroylquinic acid as biomarkers for the identification of resistant and susceptible apple cultivars to RAA and disclosed that only hydroxycinnamic acids are involved in the disease susceptibility of cultivars. In this sense, the final steps of the biosynthesis of caffeoylquinic acid (CQA) and p-coumaroylquinic acid (p-CoQA) become decisive because the isomerization of 5-CQA to 4-CQA is favored in resistant cultivars, whereas the isomerization of 5-p-CoQA to 4-p-CoQA is favored in susceptible cultivars.

Tempranillo Grape Extract in Transfersomes: A Nanoproduct with Antioxidant Activity.

Polyphenols are gaining increasing interest due to their beneficial properties to human health. Grape pomace, the by-product of wine production, is a source of these bioactive compounds. An extract from Tempranillo grape pomace was obtained and characterized qualitatively and quantitatively. The major components found were anthocyanins, flavan-3-ols, and flavonols. To improve the bioavailability of these compounds, the extract was formulated in phospholipid vesicles, namely transfersomes. Spherical unilamellar vesicles around 100 nm each were obtained. The antioxidant activity of both the extract and the transfersomes was evaluated by using colorimetric assays (i.e., DPPH, FRAP, and Folin-Ciocalteu). The cells' viability and the antioxidant activity were assessed in keratinocytes. The results showed that the extract and the transfersomes had no cytotoxic effects and exerted remarkable antioxidant activity, which was more evident in a vesicle formulation. These findings highlighted the potential of the Tempranillo grape pomace extract and the efficacy of the incorporation into phospholipid vesicles.

Stepwise strategy based on 1H-NMR fingerprinting in combination with chemometrics to determine the content of vegetable oils in olive oil mixtures.

1H NMR fingerprinting of edible oils and a set of multivariate classification and regression models organised in a decision tree is proposed as a stepwise strategy to assure the authenticity and traceability of olive oils and their declared blends with other vegetable oils (VOs). Oils of the 'virgin olive oil' and 'olive oil' categories and their mixtures with the most common VOs, i.e. sunflower, high oleic sunflower, hazelnut, avocado, soybean, corn, refined palm olein and desterolized high oleic sunflower oils, were studied. Partial least squares (PLS) discriminant analysis provided stable and robust binary classification models to identify the olive oil type and the VO in the blend. PLS regression afforded models with excellent precisions and acceptable accuracies to determine the percentage of VO in the mixture. The satisfactory performance of this approach, tested with blind samples, confirm its potential to support regulations and control bodies.

Fog reduces transpiration in tree species of the Canarian relict heath-laurel cloud forest (Garajonay National Park, Spain).

The ecophysiologic role of fog in the evergreen heath-laurel 'laurisilva' cloud forests of the Canary Islands has not been unequivocally demonstrated, although it is generally assumed that fog water is important for the survival and the distribution of this relict paleoecosystem of the North Atlantic Macaronesian archipelagos. To determine the role of fog in this ecosystem, we combined direct transpiration measurements of heath-laurel tree species, obtained with Granier's heat dissipation probes, with micrometeorological and artificial fog collection measurements carried out in a 43.7-ha watershed located in the Garajonay National Park (La Gomera, Canary Islands, Spain) over a 10-month period. Median ambient temperature spanned from 7 to 15 degrees C under foggy conditions whereas higher values, ranging from 9 to 21 degrees C, were registered during fog-free periods. Additionally, during the periods when fog water was collected, global solar radiation values were linearly related (r2=0.831) to those under fog-free conditions, such that there was a 75+/-1% reduction in median radiation in response to fog. Fog events greatly reduced median diurnal tree transpiration, with rates about 30 times lower than that during fog-free conditions and approximating the nighttime rates in both species studied (the needle-like leaf Erica arborea L. and the broadleaf Myrica faya Ait.). This large decrease in transpiration in response to fog was independent of the time of the day, tree size and species and micrometeorological status, both when expressed on a median basis and in cumulative terms for the entire 10-month measuring period. We conclude that, in contrast to the turbulent deposition of fog water droplets on the heath-laurel species, which may be regarded as a localized hydrological phenomenon that is important for high-altitude wind-exposed E. arborea trees, the cooler, wetter and shaded microenvironment provided by the cloud immersion belt represents a large-scale effect that is crucial for reducing the transpirational water loss of trees that have profligate water use, such as those of the 'laurisilva'.

Improved Thermal and Reusability Properties of Xylanase by Genipin Cross-Linking to Magnetic Chitosan Particles.

Enzymes are gradually increasingly preferred over chemical processes, but commercial enzyme applications remain limited due to their low stability and low product recovery, so the application of an immobilization technique is required for repeated use. The aims of this work were to produce stable enzyme complexes of cross-linked xylanase on magnetic chitosan, to describe some characteristics of these complexes, and to evaluate the thermal stability of the immobilized enzyme and its reusability. A xylanase was cross-linked to magnetite particles prepared by in situ co-precipitation of iron salts in a chitosan template. The effect of temperature, pH, kinetic parameters, and reusability on free and immobilized xylanase was evaluated. Magnetization, morphology, size, structural change, and thermal behavior of immobilized enzyme were described. 1.0 ± 0.1 µg of xylanase was immobilized per milligram of superparamagnetic chitosan nanoparticles via covalent bonds formed with genipin. Immobilized xylanase showed thermal, pH, and catalytic velocity improvement compared to the free enzyme and can be reused three times. Heterogeneous aggregates of 254 nm were obtained after enzyme immobilization. The immobilization protocol used in this work was successful in retaining enzyme thermal stability and could be important in using natural compounds such as Fe3O4@Chitosan@Xylanase in the harsh temperature condition of relevant industries.

Broccoli processing wastes as a source of peroxidase.

A peroxidase isozyme (BP) was purified to homogeneity from broccoli stems ( Brassica oleraceae var. maraton) discarded from industrial processing wastes. BP specific activity was 1216 ABTS [2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)] units/mg, representing 466-fold that of crude extract. BP is a monomeric glycoprotein containing 16% carbohydrates, with a molecular mass of 49 kDa and an isoelectric point close to 4.2. From kinetic data it showed a two-substrate ping-pong mechanism, and the catalytic efficiency measured as the rate-limiting step of free BP regeneration was 3.4 x 10(6) M(-1) s(-1). The ABTS K m value was 0.2 mM, which was about 20 times lower than that reported for acidic commercial horseradish peroxidase (HRP). Assessment of BP secondary structure showed 30% helical character, similar to HRP and cytochrome c peroxidase. BP lost only 25% activity after 10 min of heating at 55 degrees C and pH 6; it was stable in the pH range from 4 to 9 and showed an optimum pH of 4.6 using ABTS as substrate. BP was active on substrates normally involved in lignin biosynthesis, such as caffeic and ferulic acids, and also displayed good catechol oxidation activity in the presence of hydrogen peroxide. Reverse micellar extraction was successfully used as potential large-scale prepurification of broccoli peroxidase, achieving a purification factor of 7, with 60% activity yield. Stems from the broccoli processing industry have a high potential as an alternative for peroxidase purification.

An alternative method to estimate zero flow temperature differences for Granier's thermal dissipation technique.

Calibration of the Granier thermal dissipation technique for measuring stem sap flow in trees requires determination of the temperature difference (DeltaT) between a heated and an unheated probe when sap flow is zero (DeltaT(max)). Classically, DeltaT(max) has been estimated from the maximum predawn DeltaT, assuming that sap flow is negligible at nighttime. However, because sap flow may continue during the night, the maximum predawn DeltaT value may underestimate the true DeltaT(max). No alternative method has yet been proposed to estimate DeltaT(max) when sap flow is non-zero at night. A sensitivity analysis is presented showing that errors in DeltaT(max) may amplify through sap flux density computations in Granier's approach, such that small amounts of undetected nighttime sap flow may lead to large diurnal sap flux density errors, hence the need for a correct estimate of DeltaT(max). By rearranging Granier's original formula, an optimization method to compute DeltaT(max) from simultaneous measurements of diurnal DeltaT and micrometeorological variables, without assuming that sap flow is negligible at night, is presented. Some illustrative examples are shown for sap flow measurements carried out on individuals of Erica arborea L., which has needle-like leaves, and Myrica faya Ait., a broadleaf species. We show that, although DeltaT(max) values obtained by the proposed method may be similar in some instances to the DeltaT(max) predicted at night, in general the values differ. The procedure presented has the potential of being applied not only to Granier's method, but to other heat-based sap flow systems that require a zero flow calibration, such as the Cermák et al. (1973) heat balance method and the T-max heat pulse system of Green et al. (2003).

On-site hydrolytic enzymes production from fungal co-cultivation of Bermuda grass and corn cob.

Solid state fermentation (SSF) is used to produce industrial enzymes. The objective of this study was to use a co-culture of Aspergillus niger GS1 and Trichoderma reesei, grown on a mixture of Bermuda grass and corn cob to obtain fermented forage (FF) rich in hydrolytic enzymes, as a value added ingredient for animal feed. FPase, amylase and xylanase productivities (dry matter, DM) were 8.8, 181.4, and 42.1Ug(-1)h(-1), respectively (1U=reducing sugars released min(-1)), after 12-16h of SSF with C/N=60. Cellulose, hemicellulose and lignin decreased 1.6-, 2.7- and 1.9-fold (DM), respectively. In vitro ruminal and true digestibility of DM was improved 2.4- and 1.4-fold. Ruminal digestion of FF reduced 1.32-fold the acetate:propionate ratio, which may reduce the environmental impact of ruminants feeding. On-site hydrolytic enzymes productivity using SSF without enzymes extraction could be of economic potential for digestibility improvement in animal feed.

Monosaccharide composition and properties of a deglycosylated turnip peroxidase isozyme.

A neutral peroxidase isozyme (TP) purified from turnip (Brassica napus L. var. purple top white globe) was partially deglycosylated, using chemical and enzymatic treatment. A 32% carbohydrate removal was achieved by exposing TP to a mixture of PNGase F, O-glycosidase, NANase, GALase III and HEXase I, while m-periodate treatment removed about 88% of TP carbohydrate moiety. The glycoprotein fraction of the TP contained a relatively high mannose and fucose content (37 and 31%, w/w, respectively), 16% (w/w) galactose, and 15% (w/w) GlcNAc. Thus, the carbohydrate moiety was classified as a hybrid type. Partially deglycosylated TP had reduced activity (by 50-85%), was more susceptible to proteolysis, and showed a slight decrease in thermostability compared to the native enzyme. Circular dichroism studies strongly suggested that although the carbohydrate moiety of TP did not influence the conformation of the polypeptide backbone, its presence considerably enhanced protein conformational stability toward heat. Removal of oligosaccharide chains from TP caused a decrease in K(m) and V(max) for hydrogen peroxide. Native and chemically deglycosylated TP were similarly immunodetected by rabbit polyclonal antibodies raised against TP. The results suggest that the carbohydrate moiety of TP is important for peroxidase activity and stability.

Isolation and thermal characterization of an acidic isoperoxidase from turnip roots.

An acidic peroxidase (pI approximately 2.5) was purified from turnip roots (TAP), and its thermal properties were evaluated. TAP is a monomeric protein having a molecular weight (MW) of 49 kDa and a carbohydrate content accounting for 18% of the MW. The yield of pure TAP was relatively high ( approximately 2 mg/kg of fresh roots), with a specific activity of 1810 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) units/mg at pH 6. The activity increased 4-fold at the optimum pH (4.0) to 7250 ABTS units/mg, higher than that of most peroxidases. TAP was heat stable; heat treatment of 25 min at 60 degrees C resulted in 90% initial activity retention, whereas an activity of 20% was retained after 25 min of heating at 80 degrees C. TAP regained 85% of its original activity within 90 min of incubation at 25 degrees C, following heat treatment at 70 degrees C for 25 min. Thermal inactivation caused noticeable changes in the heme environment as evaluated by circular dichroism and visible spectrophotometry. TAP was rapidly denatured by heating in the presence of 1.0 mM ethylene glycol bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid, but the Soret band and activity were fully recovered by adding an excess of Ca(2+). This is further evidence that Ca(2+) plays an important role in the stability of TAP. The high specific activity of TAP, together with its relatively high thermal stability, has high potential for applications in which a thermally stable enzyme is required.

Scaling Erica arborea transpiration from trees up to the stand using auxiliary micrometeorological information in a wax myrtle-tree heath cloud forest (La Gomera, Canary Islands).

We investigate evapotranspiration, sap flow and top soil water content variations in a wax myrtle-tree heath ('fayal-brezal' in Spanish) cloud forest in the Garajonay National Park (La Gomera, Canary Islands) over a 1-year period. We provide transpiration estimates for one of the representative species, the shrubby needle-like Erica arborea L., present in this relict subtropical forest. An ad hoc tree up to the stand scaling method that combines the sap flow and auxiliary reference evapotranspiration data is illustrated, showing to be useful when sap flow in a limited number of trees has been monitored. Individual daily-based scaling curves of the Gompertz type were necessary to explain the observed sap flow variability in E. arborea during the 1-year period investigated (r(2) ≥ 0.953 with mode of r(2) = 0.9999). The mean daily sap flow of an E. arborea individual amounted to 8.37 ± 5.65 kg day(-1) tree(-1), with a maximum of 20.48 kg day(-1) tree(-1), yielding an annual total of 3052.89 kg tree(-1). A comparison of the computed daily transpiration with the continuous micrometeorological time series monitored in the studied plot suggested that solar radiation was the main driving force of transpiration in E. arborea (cross correlation index = 0.94). Fog may also affect tree transpiration via its reduction of radiation and temperature, such that during foggy periods the mean daily water loss estimate of E. arborea was 5.35 ± 4.30 kg day(-1) tree(-1), which sharply contrasted with the 2.4-fold average transpiration values obtained for fog-free days, i.e., 12.81 ± 4.33 kg day(-1) tree(-1). The annual water balance rendered a 288 mm year(-1) water input to the forest and evidenced the need for accurately quantifying the contribution of fog water dripping from the canopy.

Xylanolytic enzymes production by Aspergillus niger GS1 from solid-state fermentation on corn stover and their effect on ruminal digestibility

Hemicellulosic agricultural by-products such as corn stover (CS) are highly available materials which represent an opportunity to develop value added products. Native Aspergillus niger GS1 was used for solid-state fermentation (SSF) on alkali pre-treated CS (ACS) aimed to optimize xylanolytic enzymes production, and their effect on in vitro ruminal and true digestibility of ACS. Enzyme production was empirically modelled using a fractional factorial design 2(9-5), and the resulting significant factors were glucose, yeast extract and two mineral salts, which were arranged in a Draper-Lin optimization design at two levels. Predicted optimum xylanolytic activity of 33.6 U (mg protein)-1 was achieved at 48 hrs of SSF, and was validated by confirmatory experiments. ACS was incubated with a semipurified enzymatic extract (EE) showing a xylanolytic activity of 1600 U kg-1 dry ACS for 12 hrs before exposure to cow's ruminal liquid for 72 hrs, which led to 5 percent and 10 percent increase of in vitro ruminal and true digestibility, respectively. CS is a readily available by-product in different regions which after alkaline treatment and partial hydrolysis with the EE, may be advantageously used as supplement for ruminant feed.

Morbimortalidad materna por aborto / Maternal morbimortality due to abortion

Antecedentes: Diversas técnicas efectuadas para provocar el aborto se asocian a complicaciones y a las posibilidades y alteraciones que inciden en la probabilidades de la mujer en este trance de morir y/o enfermar. Materiales y métodos: A 106 mujeres admitidas en la Maternidad Nuestra Señora de la Altagracia en Santo Domingo, República Dominicana, con el diagnóstico de aborto incompleto le aplicamos un instrumento para la recolección de información respecto a edad, escolaridad, estado civil, ocupación, procedencia, mortalidad, morbilidad, método abortivo y complicaciones. Resultados: El 39.6// de las pacientes tenían de 15 a 20 años de edad y 32.1// de 21 a 25 años. El 56.6// vivían en unión libre y 30.2// eran solteras. Un 52.8// procedían de la zona urbana y 47.2// de la zona rural. El 82.0// eran estudiantes o se dedicaban a quehaceres domésticos. El 56.5// tenían estudios secundarios y/o universitarios, 36.8// alcanzaron estudios primarios o intermedios. En 76.4// la condiciones mórbidas eran sangrado y dolor y 23.6// presentaban ademas de estas condiciones fiebre, mareos e infección. Un 78.3// no presentó complicación alguna, 11.3// hemorragia y un 8.5// infección. Registramos un caso de perforación uterina y un caso de laceración del cervix. Un 63.2// habían usado anticonceptivos orales y en 29.3// el aborto era espontáneo