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Aims: Some bacterial responses to oxidative stress also diminish antibiotic susceptibility; also, some antibiotics do increase oxidative stress within bacterial cells. Linkage or cross-resistance to prooxidants and antibiotics could facilitate the selection of antibiotic resistance and/or virulence. We made this survey in order to detect this possible linkage in Escherichia coli isolates. Methodology: The susceptibility of 102 E. coli clinical (causative of urinary or gastrointestinal infections) and environmental (rural or urban dust) isolates towards paraquat, H2O2, and antibiotics was measured using disc assays. Catalase and superoxide-dismutase (SOD) activities were measured. Results: Susceptibility to prooxidants was similar across isolates of all four sources, but urinary and urban dust isolates were more resistant to antibiotics. H2O2 "resistant" organisms had more antibiotic resistance phenotypes, particularly towards sulfadiazine and tetracycline. Paraquat "resistance" seems associated to beta-lactam resistance; but paraquat "susceptibility" seems associated to resistance towards chloramphenicol, gentamicin, ciprofloxacin and nitrofurantoin. Prooxidant disc assays correlate to catalase and superoxide-dismutase activities. A weak relationship H2O2/antibiotic-resistance, but not superoxide/antibiotic-resistance, is suggested. Conclusion: Overall, antibiotics exerting their action through oxidative stress, do not seem to have resulted in the co-selection of oxidative stress resistance, or vice versa. However, a possible link between resistance to some antibiotics and to H2O2 might contribute to co-selection between these two chemical insults.

Objetivo: comparar el gasto en alimentos de baja calidad nutricional (ABCN), de los hogares de Sonora y a nivel nacional de acuerdo con su decil de ingreso. Metodología: con base en la llamada Encuesta Nacional de Ingresos y Gastos de los Hogares (ENIGH 2016-2018) y mediante la técnica estadística ANOVA de un solo factor, se busca determinar las diferencias del gasto de los hogares entre los distintos deciles socioeconómicos. Para la selección de alimentos de baja calidad nutricional se utilizaron los criterios propuestos por Darmon y Drewnosky (2008), y el aporte energético de los alimentos de Ledwike et al (2006). Resultados: se encontraron diferencias significativas que muestran que los hogares con deciles bajos gastan más en ABCN que los deciles altos. Sonora tiene un consumo alto en alimentos como galletas y botanas. En tanto el gasto en refrescos es homogéneo en todos los deciles socioeconómicos. Limitaciones: la ENIGH solo permite estudiar el gasto en productos energéticos, no permite establecer conclusiones sobre cuantas kilocalorías consume la población. Conclusiones: el gasto en alimentos de baja calidad nutricional es significativo entre los hogares de bajos ingresos y se adecua a los resultados encontrados por Darmon y Drewnosky en otros países europeos.

The effects of the carbonic maceration and conventional winemaking on the volatile composition and aromatic sensory characteristics of Tempranillo Blanco wines were studied for the first time, during three consecutive vintages. Relationships between instrumental (volatiles) and sensory variables were analyzed applying partial least squares regression (PLS). Carbonic macerated wines had higher contents of alcohols and carbonyl compounds, yet lower concentrations of C6 alcohols and volatile acids than wines conventionally produced. The Odor Activity Values (OAV) exhibited an increase in wines when carbonic maceration was applied. According to the geometric mean (% GM) obtained from aroma descriptors the effect of the winemaking process was significant for seed fruit, ripe fruit and floral notes. When subjected to PLS the data from the instrumental analysis yielded a satisfactory model for the prediction of aroma descriptors in this set of wines.

Vitis vinífera L. cv. Maturana Blanca is an autochthonous minor variety recently recovered in the Rioja Qualified Denomination of Origin (D.O.Ca Rioja, Spain) for the production of monovarietal white wines with singular and differentiated characteristics. In this paper, Maturana Blanca wines made with different technologies were analyzed by sensory analysis and aromatic profile by gas chromatography-mass detector. Maturana Blanca wines were characterized by low pH, high acidity, and yellowish tonalities. The compounds that most influenced the aroma of Maturana Blanca wines were those related to fruity (acetates and ethyl esters), floral aromas (2-phenylethanol), and spicy notes (γ-decalactone). These wines were mainly characterized by volatile compounds of fruity aromas of banana and apple. The use of pre-fermentative maceration increased the concentration of ethyl esters and acetates and produced wines with higher odor activity values, indicating a greater aromatic intensity. The aromatic profile of Maturana Blanca wines fermented in oak barrels showed a greater complexity as they were also characterized by the presence of important amounts of furfural, whiskey lactone, and eugenol. The sensory analysis confirmed the results obtained in the aromatic analysis, and described the wines as fresh and balanced in mouth, with notes of acidity and medium to high persistence. These results will contribute to a better knowledge of this white variety.

This paper studied the effect on the molecular weight and polysaccharide composition of musts and wines of the application of high-power ultrasound (US) at 20 and 28 kHz on crushed grapes. Two different pomace maceration times (short and mid) were tested for sonicated and control vinifications. A long pomace maceration time was also tested for non-treated wines. In must samples, US significantly increased the content of monosaccharides and polysaccharides rich in arabinose and galactose (PRAG), and the average molecular weight of smaller PRAG, mannoproteins (MP) or mannans. The 28 kHz had a major effect on most wine monosaccharides and grape polysaccharides. The wine obtained from sonicated grapes at 28 kHz and with mid maceration had higher rhamnogalacturonans type II and PRAG content than its control, and closer polysaccharide and monosaccharide content to long maceration control wines. No significant differences were obtained in the MP content between sonicated and control wines., This research was funded by the Ministerio de Ciencia, Innovación y Universidades from the Spanish Government and Feder Funds, grant number RTI2018-093869-B-C21.

The aim of this paper is to optimize a method to recover polysaccharides from white grape pomace (non-fermented), the main waste by-product of the food industry. Different conditions are tested and the polysaccharides extracted are analyzed by high performance size exclusion chromatography with refractive index detector (HPSEC-RID) and gas chromatography with mass detector (GC-MS). The extraction solvent did not show a significant effect on the polysaccharide extraction, acid pH yielded to higher efficiencies, and longer extraction times extracted more smaller polysaccharides (≤5.4 kg mol). The highest efficiencies were obtained with both solvents at pH 1 and 1:4 solid to liquid ratio. The optimum conditions selected (TA as solvent of extraction, 2.5 g L solvent concentration, pH = 1, 1:4 solid to liquid ratio, and 18 h of extraction time) allow the extraction of polysaccharides rich in arabinose and galactose, rhamnogalacturonans, homogalacturonans and glucosyl polysaccharides, under efficient and food-safe conditions., The authors would like to thank the Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA) and the Ministerio de Ciencia e Innovación for the funding provided for this study through the project RTA2017-00005-C02-02.

The aim of this work was to study oenological techniques to obtain adequate base wine for red sparkling wine elaboration. Four winemaking techniques were carried out: pre-fermentative cold maceration with dry ice and delestage with premature grapes; and sugar reduction in must and partial dealcoholisation of wine with mature grapes. Their effect on oenological parameters, volatile composition, foam and sensory characteristics was valuated. Reduction of sugar content and partial dealcoholisation allow obtaining base wines with more adequate alcohol content. No differences were found between the oenological parameters during the ageing time. Pre-fermentative cold maceration and partial dealcoholisation had a greater influence on the volatile composition of the base and red sparkling wines. Oenological technique did not affect the foam instrumental parameters. Red sparkling wines from premature grapes showed higher vegetal aromas, and pre-fermentative cold maceration the best foam sensory descriptors., The authors would like to thank the ‘Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria’ (INIA) for the funding provided for this study through the project RTA2012-092-C02-01 (with FEDER funds). M. G-L. thanks the Comunidad Autónoma de La Rioja and Universidad de La Rioja for her FPI-UR-CAR grant.

The water vapor permeability (WVP) and hardness of biodegradable nanocomposite films based on glutelin from Jatropha curcas L. were determined and the results were analyzed by response surface meth...

Finding new alternatives to traditional live preys such as Artemia and rotifers, which do not always promote optimal fish growth and survival, is required for the successful aquaculture of highly specialized predatory species, including seahorses. The present study assessed the nutritional value of an interesting marine amphipod (Parhyale hawaiensis), and evaluates through a feeding trial its potential use as a natural prey for 10-months lined seahorse, Hippocampus erectus. P. hawaiensis showed high levels of valuable lipids (20.4–26.7% on dry matter basis) and polyunsaturated fatty acids (PUFAs) ( 26.4–41% of total FAs), including the long-chain PUFAs (LC-PUFAs) arachidonic acid (ARA) (2.9–7.7%), eicosapentaenoic acid (EPA) (4.3–6.5%) and docosahexaenoic acid (DHA) (2.1–6.2%). A comparison between wild-captured and cultured amphipods revealed a significant improvement of the amphipod FA profile in terms of DHA%, total omega-3 (n3) FAs and n3/n6 ratio when employing both a conventional amphipod culture based on a commercial shrimp diet, and, to a lesser extent, a large (3,500 L) biofloc system. Seahorses fed with frozen/wild amphipods, either singly or in combination with Artemia enriched with Super Selco® (INVE Aquaculture, Belgium) for 57 days, substantially improved seahorse growth and FA profiles in terms of ARA, EPA and DHA%, including indices associated to marine sources, such as Σn3 and n3/n6, compared to a diet based solely on enriched Artemia. These results support the use of marine amphipods as an alternative food organism for juvenile H. erectus and suggest a potential use for general marine aquaculture.

Consumer concern about the relationship between food and health has generated a growing interest in functional foods. Conjugated linoleic acid (CLA) has been associated with several beneficial health properties. Therefore, this study reports the design of co-cultures containing CLA-producing strains and their performance in MRS and in skim milk. The four designed co-cultures were tested under different conditions. In MRS, co-culture 2 (containing autochthonous Lactococcus lactis strains and Lactobacillus plantarum TAUL 1588) was the highest CLA-producer (98.01 µg/mL) after 48 h incubation. In milk, the highest total CLA production (56.51 µg/mL) was reached with co-culture 4 (containing autochthonous Lactococcus lactis, Lactobacillus plantarum TAUL 1588 and Lactobacillus casei SS 1644) after 72 h. These results open the way for the use of co-cultures containing CLA-producing strains to manufacture cheese and obtain functional dairy products rich in bioactive fatty acids.

BACKGROUND: Verdejo and Tempranillo are traditional varieties for producing still wines; however, they could provide an alternative for the manufacturing of sparkling wines. Sparkling wines were elaborated by the traditional method, followed by ageing on lees for 9 months. A study on the changes that take place in polysaccharides, oligosaccharides and nitrogenous compounds during the ageing on lees of Tempranillo and Verdejo sparkling wines has been undertaken. RESULTS: Mannoproteins and the glucose residue of oligosaccharides were the major carbohydrates detected in all vinification stages. Yeast polysaccharides and glucan-like structures of the oligosaccharides increased after 3 months of ageing. The evolution of yeast polysaccharides and the composition of PRAG-like structure were different among grape varieties. A decrease in amino acids and biogenic amines was observed during the ageing. The contents of polysaccharides, oligosaccharides and nitrogenous compound were significantly higher in Tempranillo than in Verdejo sparkling wines at the end of the ageing period. CONCLUSION: Polysaccharides and oligosaccharides from yeast were more significant autolysis markers of sparkling wines than the nitrogenous compounds. Our data suggest a potential cultivar effect on the evolution of yeast polysaccharides and on the composition of PRAG, which may influence the physico-chemical and sensory properties of sparkling wines.

In this study, 85 strains of lactic acid bacteria isolated from artisanal cheeses were screened to determine their capacity to synthesise bioactive compounds. Four Lactobacillus plantarum and two Lactobacillus casei subsp. casei strains were capable of synthesizing conjugated linoleic acid (CLA). The highest level of CLA formed in MRS after 48 h incubation was 19.26 µg/mL. Six Lactobacillus brevis and four Lactococcus lactis subp. lactis strains produced gamma-aminobutyric acid (GABA) and the highest concentration (2524.05 µg/mL) was found after 72 h incubation. The detection of genes encoding linoleate isomerase or glutamate decarboxylase system could be suitable for use as screening method of CLA or GABA-producer strains, although this should be studied in a larger number of strains. Our findings suggest that the above strains are potential candidates for the design of starter cultures with the capacity to generate bioactive compounds, offering new possibilities for the manufacture of functional dairy products.

Polysaccharides are the main macromolecules of colloidal nature in wines, and play a fundamental role in the technological properties and organoleptic characteristics of the wines. The role of the different wine polysaccharides will depend on their quantity but also on their chemical composition, molecular structure and origin. Wine polysaccharides originate from grapes and yeast acting during the winemaking. The main polysaccharides present in wines can be grouped into three major families: (i) polysaccharides rich in arabinose and galactose (PRAG), (ii) polysaccharides rich in rhamnogalacturonans (RG-I and RG-II), which both come from the pectocellulosic cell walls of grape berries, and (iii) mannoproteins (MP) released by yeasts. This paper describes the origin, structure and role of the different wine polysaccharide families through a bibliographic revision of their origin and extraction into the wines, as well as their technological and sensory properties.

Mechanisms contributing to pulmonary and systemic injury induced by high tidal volume (VT) mechanical ventilation are not well known. We tested the hypothesis that increased peroxynitrite formation is involved in organ injury and dysfunction induced by mechanical ventilation. Male Sprague-Dawley rats were subject to low- (VT, 9 mL/kg; positive end-expiratory pressure, 5 cmH2O) or high- (VT, 25 mL/kg; positive end-expiratory pressure, 0 cmH2O) VT mechanical ventilation for 120 min, and received 1 of 3 treatments: 3-aminobenzamide (3-AB, 10 mg/kg, intravenous, a poly adenosine diphosphate ribose polymerase [PARP] inhibitor), or the metalloporphyrin manganese(III) tetrakis(1-methyl-4-pyridyl)porphyrin (MnTMPyP, 5 mg/kg intravenous, a peroxynitrite scavenger), or no treatment (control group), 30 min before starting the mechanical ventilation protocol (n = 8 per group, 6 treatment groups). We measured mean arterial pressure, peak inspiratory airway pressure, blood chemistry, and gas exchange. Oxidation (fluorescence for oxidized dihydroethidium), protein nitration (immunofluorescence and Western blot for 3-nitrotyrosine), PARP protein (Western blot) and gene expression of the nitric oxide (NO) synthase (NOS) isoforms (quantitative real-time reverse transcription polymerase chain reaction) were measured in lung and vascular tissue. Lung injury was quantified by light microscopy. High-VT mechanical ventilation was associated with hypotension, increased peak inspiratory airway pressure, worsened oxygenation; oxidation and protein nitration in lung and aortic tissue; increased PARP protein in lung; up-regulation of NOS isoforms in lung tissue; signs of diffuse alveolar damage at histological examination. Treatment with 3AB or MnTMPyP attenuated the high-VT mechanical ventilation-induced changes in pulmonary and cardiovascular function; down-regulated the expression of NOS1, NOS2, and NOS3; decreased oxidation and nitration in lung and aortic tissue; and attenuated histological changes. Increased peroxynitrite formation is involved in mechanical ventilation-induced pulmonary and vascular dysfunction. Sin financiación 3.048 JCR (2015) Q1, 33/200 Surgery; Q2, 12/33 Critical care medicine, 29/70 Hematology, 23/63 Peripheral vascular disease UEM

This study was performed to determine how two of the most important isoflavones, genistein and daidzein, affect the gonadal axis in male prepuberal rats. One hundred and seventy-five prepuberal male Wistar rats were allocated into seven groups: one control group and six experimental groups that were orally administered a high or low dose of genistein, daidzein or a mixture of both. Testosterone determination was assayed by EIA. The testes and body weights were measured, and the histology of the epididymis with the sperm content and epididymal sperm count were evaluated. In the control group, we observed an increase in the serum testosterone levels (>2.5 ng ml(-1) ) at the third week (52 days), which corresponded to the onset of puberty in these rats. The same increase in serum testosterone levels was observed at the fourth week in rats that received low doses of isoflavones; therefore, we concluded that the onset of puberty was delayed. At high doses, there was no significant increase in testosterone levels, which could be related to the fact that these male rats did not reach puberty. These findings were supported by the results obtained from the analysis of the epididymal content as well as the testes/body weight ratio.

Traditional cheeses are an important reservoir of microbial diversity that can have important biotechnological applications, especially with a view to improving the characteristics unique to each type of cheese, and in this respect, starter cultures consisting of autochthonous lactic acid bacteria strains are of particular interest. In the present study, we investigated lactic acid bacteria population diversity in San Simon da Costa cheese (PDO, Galicia, Spain) and found a predominance of the genus Lactobacillus, which by the end of ripening accounted for 78% of the strains isolated in Rogosa agar, around 40% of those in M17 agar and about 10% of those in MSE agar. The main species of lactic acid bacteria identified were Lactococcus lactis subsp. lactis, Lactobacillus casei subsp. casei, Lb. paracasei subsp. paracasei, Leuconostoc mesenteroides and Enterococcus faecalis. Virtually all the strains studied from a technological point of view yielded more than or equal to 0.24 g 100 mL-1 lactic acid. Lactococcus lactis subsp. lactis (SS 194), Lactobacillus paracasei (SS 1695 and SS 1689) and Enterococcus faecalis (SS 1378 and SS 1449) strains exhibited the greatest proteolytic capacity. Based on the overall technological aptitude of the tested strains, we can propose starter cultures and co-cultures that include different combinations of previous strains with a view to manufacturing San Simon da Costa cheese from pasteurised milk.

CIGB 247 is a novel cancer therapeutic vaccine based on human vascular endothelial growth factor (VEGF) variant molecule as antigen, in combination with a bacterial adjuvant. This vaccine candidate has previously demonstrated efficacy and safety in mice, rats, rabbits and non-human primates. In the present study we evaluated the effects on the clinical, hematological and biochemical parameters of CIGB 247 vaccine in Chlorocebus aethiops monkeys. Three groups of monkeys were immunized with three doses of vaccine formulation to measure physiological values of clinical, hematological and serum biochemical parameters. Monkeys' body weight and temperature were kept stable and close to standard values throughout the study. Variations in the levels of red blood cells and hemoglobin were observed among the different groups for all injected doses, but these hematological parameters recovered normal values at the end of the study. On the other hand, biochemical parameters such as the total bilirubin and total protein counts showed variations along the study, while they were not associated with the test substance. In summary, no negative effects on clinical, hematological and biochemical parameters were detected. Together, our results put forward the potential and support the safety of the CIGB 247 vaccine candidate for use in clinical applications. The data presented here can be used to estimate a human dosing regimen from preclinical data.

BACKGROUND Physicochemical characteristics, proteolysis and lipolysis were studied throughout the ripening of eight batches of a traditional Spanish variety made from raw cow's milk, in order to establish a basis for its industrial production. RESULTS The main compositional characteristics of this cheese after 60 days of ripening were its high proportion of total solids (TS; 752 g kg−1 of cheese), an average content of protein (452.8 g kg−1 TS) and fat (475.1 g kg−1 TS) and the presence of residual lactose (12.5 g kg−1 TS). Its pH value (4.04) was extremely low. Phosphorus (5.13 g kg−1 TS) and sodium (8.29 g kg−1 TS) were the most abundant mineral elements in cheese, whereas calcium levels (1.92 g kg−1 TS) were very low. Proteolysis extension and depth were very low, which resulted in almost zero degradation of αs1- and β-casein. Fat acidity increased during ripening, reaching final values of 50.1 mg KOH kg−1 of fat. The main free fatty acid was C16:0, followed by C18:1 and C14:0. CONCLUSION These results suggest that this variety undergoes a limited proteolysis and moderate lipolysis during ripening. The low pH, low calcium content and limited proteolysis led to a crumbly texture in this cheese variety. © 2014 Society of Chemical Industry

Isoflavones are the most common phytoestrogens found in human diets. However, it is still not clear whether isoflavones have effects on the reproductive and the endocrine systems under normal dietary intake and overdose. The aim of this study was to determine how the most important isoflavones, genistein and daidzein, affect androgen and glucorticoid levels on male prepuberal rats. A hundred and seventy-five 30-day-old male Wistar rats were dosed orally by stomach tube every day for 35 days, with saline solution, low and high doses of genistein, daidzein and a mixture of both. Serum samples were analysed by an enzyme immunoassay for hormone determinations. In control group, there was a peak of testosterone (T) and dihydrotestosterone levels associated to the onset of puberty, at the third week. However, in low-dose groups, the same peak was found at the fourth week (p < 0.05), indicating a delay in the onset of puberty in these groups. Moreover, high doses groups serum androgen levels were significantly lower (p < 0.05) than the control group from the first week until fifth week. This fact was supported by a epididymal histological analysis that indicate in low doses there were several content of spermatozoa at fourth week and in high doses there were few content of spermatozoa. Besides, corticosterone levels followed the same pattern of androgens in all groups. We can conclude that oral administration of isoflavones in male rats decreased the secretion of androgens and glucocorticoids causing a delay in the onset of puberty and may cause physiological and developmental problems.

[EN] The relationship between cell wall composition and extractability of anthocyanins from red grape skins was assessed in Tempranillo grape samples harvested at three stages of ripening (pre-harvest, harvest and over-ripening) and three different contents of soluble solids (22, 24 and 26 !Brix) within each stage. Cell wall material was isolated and analysed in order to determine cellulose, lignin, non-cellulosic poly- saccharides, protein, total polyphenols index and the degree of esterification of pectins. Results showed the influence of ripeness degree and contents of soluble solids on cell wall composition. Furthermore, principal components analysis was applied to the obtained data set in order to establish relationships between cell wall composition and extractability of anthocyanins. Total insoluble material exhibits the biggest opposition to anthocyanin extraction, while the highest amounts of cellulose, rhamnogalacturonans-II and polyphenols were positively correlated with anthocyanin extraction. Moreover, multiple linear regression was performed to assess the influence of the cell wall composition on the extraction of anthocyanin compounds. A model connecting cell wall composition and anthocyanin extractabilities was built, explaining 96.2% of the observed variability.

The aim of this work was to focus on the study of the volatile composition of white and rosé base wines elaborated from different autochthonous grape varieties in Spain (Verdejo,Viura, Malvasía, Albarín, Godello,PrietoPicudo and Garnacha) and the evolution of the sparkling wines elaborated from them following the “champenoise” method. The amino acids and biogenic amines were also studied. Multivariate analyses (factorial and stepwise discriminant) were carried out to study the influence of these compounds all together. Base and sparkling wines from Albarín, Verdejo, Godello and PrietoPicudo were the richest in most of the volatile compounds analyzed, especially of ethyl esters and alcohol acetates, compounds that contribute to the fruity aroma of wines. During the aging on lees in the bottle of sparkling wines, an increase in branched ethyl esters, ethyl lactate and γ-butyrolactone, and a decrease in terpenes (mainly citronellol and geraniol) were observed. In general, the levels of biogenic amines in all the sparkling wines studied were very low, which does not represent a negative effect on their quality and safety., The authors would like to thank the “Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria” (INIA) for the funding provided for this study through the project RTA2009-029-C02 (with FEDER funds). L. Martínez also thanks to La Rioja Government for the financing of her pre-doctoral fellowship.

The effect of egg albumin fining, progressive clarification and cross-flow microfiltration on the polysaccharide and proanthocyanidin composition of four red varietal wines was studied in this work. Discriminant analyses were applied to achieve a possible differentiation of the wines according to treatment or grape variety. Egg albumin fining did no produce a significant decrease in the content of wine polysaccharides. Progressive clarification caused a significant reduction of mannoproteins, homogalacturonans and polysaccharides rich in arabinose and galactose in Graciano wines. However, both treatments reduced the total content of proanthocyanidins in all varietal wines. Cross-flow microfiltration produced the highest retention of polysaccharides and proanthocyanidins in all the wines, mainly polysaccharides rich in arabinose and galactose, yeast mannoproteins and highly polymerized phenols. Polysaccharides rich in arabinose and mannoproteins were more retained on the ceramic membrane than polysaccharides rich in galactose and proanthocyanidins. Discriminant analyses allowed a clear differentiation of cross-flow microfiltrated wines from the rest of the wines.

The foam of a sparkling wine is a key parameter of its quality, and the main characteristic differentiating sparkling wines from the so-called still wines. Both foam formation and duration are directly related to the chemical composition of sparkling wines. This chapter reviews the most recent studies made to determine the influence of chemical compounds on the foamability and foam stability of sparkling wines. Foam properties of sparkling wines are ruled by a large number of molecules, but some compounds seem to be more relevant than others to explain their behavior. The content of total amino acids, polysaccharides, anthocyanins, coumaric acid, and isorhamnetin showed high correlation values with foam quality parameters. The alcohol content and the concentration of acid polysaccharides, proanthocyanidins and free SO2 are the factors which most negatively affect foam quality. A recent study, by means of prediction models, has concluded that the different forms of malvidin show the highest influence on the foamability parameters in rosé sparkling wines, followed by amino acid compounds, while foam stability model was only predicted by polysaccharides rich in arabinose and galactose. These research findings provide industry with a better understanding of the compositional factors influencing the foam quality of sparkling wines.

This paper studied how grape maturity affected complex carbohydrate composition during red sparkling wine making and wine aging. Grape ripening stage (premature and mature grapes) showed a significant impact on the content, composition, and evolution of polysaccharides and oligosaccharides of sparkling wines. Polysaccharides rich in arabinose and galactose, mannoproteins, rhamnogalacturonans II, and oligosaccharides in base wines increased with maturity. For both maturity stages, polysaccharides rich in arabinose and galactose, and the glucuronic acid glycosyl residue of the oligosaccharides were the major carbohydrates detected in all vinification stages. The total glycosyl content of oligosaccharides decreased during the whole period of aging on yeast lees. The reduction of polysaccharides rich in arabinose and galactose and rhamnogalacturonans type II during the aging was more pronounced in mature samples. To our knowledge, this is the first report of the polysaccharide and oligosaccharide composition of red sparkling wines., The authors would like to thank the Spanish Ministry of Economy and Competitiveness and the “Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria” (INIA) for the funding provided for this study through project RTA2012-00092-C02-02 (with FEDER funds). L. Martínez-Lapuente also thanks the La Rioja Government for the financing of her predoctoral fellowship.

Spanish grape varieties that have been traditionally used to produce still wines were examined for their potential to make white and rose sparkling wines. Sparkling wines manufactured from Vitis vinifera cv. Verdejo, Viura, Malvasia, Albarin, Godello, Garnacha, and Prieto varieties were examined for sensory attributes and for the evolution of monomeric and polymeric phenolics during different stages of winemaking and aging. Stabilization and clarification of the base wines significantly decreased the concentrations of anthocyanins and proanthocyanidins. During the initial months of aging on yeast lees, all types of polyphenols decreased, although some were released back into the wine during the final months. Garnacha rose and Albarin white wines had high phenolic potential. Garnacha rose sparkling wines had particularly high hydroxycinnamic acid concentrations, while Prieto Picudo rose sparkling wines had the highest color intensity and anthocyanin concentrations. Among white sparkling wines, Albarin had the most catechin, proanthocyanidins, and, together with Viura, hydroxycinnamates. In sensory profiling, Prieto Picudo had more visual color intensity, red tones, olfactory intensity, freshness sensations, and foam quality than Garnacha wines. Albarin and Verdejo had more visual color and olfactory intensity than the other white wines, and Verdejo had better foam quality. Prieto Picudo, Albarin, and Verdejo were the most promising varieties for the production of high-quality sparkling wines.

Forty-one strains of Geotrichum candidum isolated from Armada cheese, Sobado variety, were screened for their enzymatic activities, including proteolytic and lipolytic activities and aminopeptidase activity. The highest extracellular proteolytic activity was detected for 8 strains with values ranging between 2.086 and 4.685 mM Gly L −1 of milk. Extracellular lipolytic activity was high for all but one of the G. candidum strains, with values ranging between 67 and 131 μmol oleic mL −1 . Cell-bound lipase activity showed values which were considerably lower than those for extracellular activity, ranging between 32.50 and 42.50 μmol oleic mL −1 and falling below 20 μmol oleic mL −1 in 28 strains. Aminopeptidase activity was not very high in the cell-free extract (CFE) of any of the strains, showing the highest values toward the substrate Lys- p -nitroanilide, with levels of activity ranging between 6.12 and 10.05 UE mg −1 protein in 8 strains. In accordance with the results obtained, four G. candidum strains were selected as co-cultures in order to study their role in the ripening of a semi-hard goat’s milk cheese.

The mechanisms contributing to sepsis vascular dysfunction are not well known. We tested the hypothesis that peroxynitrite scavenging ameliorates sepsis-induced macrovascular and microvascular dysfunction. Male Sprague-Dawley rats were killed 48 h after cecal ligation (n = 15) and puncture or sham procedure (n = 15). Their aortas and mesenteric vessels were mounted in organ baths for isometric tension recording. We studied contraction in resting vessels (norepinephrine 1 nM-10 μM and 10 nM-10 μM) and endothelium-dependent relaxation (acetylcholine, 10 nM-10 μM and 1 nM-10 μM) for aortas and microvessels, respectively. Vascular rings were preincubated for 30 min with the superoxide scavenger Cu-Zn-superoxide dismutase (SOD) (100 U/mL), the SOD mimetic and peroxynitrite scavenger tempol (10 M), the NO synthase inhibitor N-nitro-l-arginine methyl ester (10 M), or the peroxynitrite decomposition catalyst manganese tetrakis(4-N-methylpyridyl)porphyrin (MnTMPyP) (10 M). Fluorescence to 3-nitrotyrosine, oxidized dihydroethidium, and NOS2 was assessed in vascular tissue. Vascular NOS2, endothelial nitric oxide synthase (NOS1), NADPH-oxidase-1 (NOX-1), and SOD expression was analyzed by reverse transcription-polymerase chain reaction. Sepsis induced (i) in macrovessels, impairment of norepinephrine-induced contractions; (ii) in microvessels, impairment in norepinephrine-induced contractions and acetylcholine-induced relaxations; (iii) aortic and microvascular tissue increased reactivity to 3-nitrotyrosine, oxidized dihydroethidium, NOS2, and increased expression of NOS2, as well as increased expression of NOX-1 in microvascular tissue. Contractile responses in aortic and microvascular rings improved by ex vivo treatment with MnTMPyP and tempol, whereas vascular relaxation in microvessels improved only with MnTMPyP. Peroxynitrite scavenging protects from vascular dysfunction in sepsis.

Psoriasis is a chronic autoimmune systemic disease with an approximate incidence of 2% worldwide, it is commonly characterized by squamous lesions on the skin that present the typical pain, stinging, and bleeding associated with an inflammatory response. In this work, poly(methyl vinyl ether-alt-maleic ethyl monoester) (PMVEMA-ES) nanofibers have been designed as a delivery vehicle for three therapeutic agents with palliative properties for the symptoms of this disease (salicylic acid, methyl salicylate, and capsaicin). For such a task, the production of these nanofibers by means of the electrospinning technique has been optimized. Their morphology and size have been characterized by optical microscopy and scanning electron microscopy (SEM). By selecting the optimal conditions to achieve the smallest and most uniform nanofibers, approximate diameters of up to 800&ndash, 900 nm were obtained. It was also determined that the therapeutic agents that were used were encapsulated with high efficiency. The analysis of their stability over time by GC-MS showed no significant losses of the encapsulated compounds 15 days after their preparation, except in the case of methyl salicylate. Likewise, it was demonstrated that the therapeutic compounds that were encapsulated conserved, and even improved, their capacity to activate the transient receptor potential cation channel 1 (TRPV1) channel, which has been associated with the formation of psoriatic lesions.

Exposure to particulate matter (PM) induces inflammatory cytokines. In the present study, we evaluated the secretion of IL-6 and IL-8 by an airway cell line exposed to PM with a mean aerodynamic size equal to or less than 10 or 2.5 microm (PM10 and PM2.5, respectively) collected in Mexico City, using a modified high-volume sampling method avoiding the use of solvents or introducing membrane components into the samples. PM was collected on cellulose-nitrate (CN) membranes modified for collection on high-volume samplers. Composition of the particles was evaluated by particle-induced X-ray emission (PIXE) and scanning electron microscopy. The particles (10-160 microg/cm2) were tested on Calu-3 cells. Control cultures were exposed to LPS (10 ng/mL to 100 microg/mL) or silica (10-160 microg/cm2). IL-6 and IL-8 secretions were evaluated by ELISA. An average of 10 mg of PM was recovered form each cellulose-nitrate filter. No evidence of contamination from the filter was found. Cells exposed to PM10 presented an increase in the secretion of IL-6 (up to 400%), while IL-8 decreased (from 40% to levels below the detection limit). A similar but weaker effect was observed with PM2.5. In conclusion, our modified sampling method provides a large amount of urban PM free of membrane contamination. The urban particles induce a decrease in IL-8 secretion that contrasts with the LPS and silica effects. These results suggest that the regulation of IL-8 expression is different for urban particles (complex mixture containing combustion-related particles, soil and biologic components) than for biogenic compounds or pure mineral particles.

The aim of this work was to study the antimicrobial activity of 395 strains of lactic acid bacteria isolated from Genestoso cheese throughout manufacture and ripening. These were: Lactobacillus (137 strains), Lactococcus (125 strains), Leuconostoc (58 strains) and Enterococcus (75 strains). After the extract had been neutralized and treated with catalase to exclude the action of hydrogen peroxide, 13 lactococci, three lactobacilli, three leuconostocs and five enterococci continued to show antimicrobial activity against one or more of the reference varieties used as indicator strains (Enterococcus faecalis CECT 481, Staphylococcus aureus CECT 240, Lactobacillus plantarum CECT 748, Clostridium tyrobutyricum CECT 4011 and Listeria monocytogenes CECT 4031). The 24 strains of lactic acid bacteria showing antimicrobial activity after excluding the action of hydrogen peroxide were identified at species level by genetic methods. The majority of these strains were isolated from the batch ripened in autumn (37.5% of strains) and the batch ripened in spring (29.2% of 15 strains) manufactured by the same cheesemaker and mainly during manufacture and the first days of ripening.

Thirty-one lactic acid bacteria isolated from Armada cheese and previously selected in accordance with their technological properties, were screened for antimicrobial activity one against another. Four strains showed inhibitory activity against 14 strains when tested by well diffusion assay after the effects of organic acid and hydrogen peroxide were eliminated. Extracts of the strains did not show inhibitory activity after treatment with proteinase K, trypsin, α-chymotrypsin or pepsin. None of the strains showed antimicrobial activity against several pathogenic and spoilage reference strains. Eighteen representative strains were tested for their antibiotic resistance. None of the strains were totally susceptible to all antibiotics tested and multiple resistance was observed. Most of the tested strains were resistant to cefotoxin, oxacillin, vancomycin, teicoplanin, nitrofurantoin and trimethoprim.

Background and Purpose— Elevated plasma levels of homocysteine are associated with an increased risk of deep-vein thrombosis. Through a case–control study, we examined the potential association among homocysteine, folate and vitamin B 12 levels, and the common C677→T mutation in the methylene tetrahydrofolate reductase ( MTHFR ) gene in patients with cerebral venous thrombosis (CVT). Methods— Forty-five patients with CVT and 90 control subjects were studied. Plasma levels of homocysteine (fasting and after methionine load), folate, and vitamin B 12 were measured. Genotyping of the MTHFR gene was also performed. The estimated risk of CVT associated with hyperhomocysteinemia, low vitamin levels, and MTHFR mutation were expressed as odds ratio (OR) and its 95% CI (crude and after adjusting by other independent variables). Results— The adjusted OR for CVT associated with high (>90th percentile) fasting levels of homocysteine was 4.6 (1.6 to 12.8). The association between low plasma folate values (MTHFR mutation in patients with CVT (22% versus 10%), but it was not statistically significant ( P =0.098). Patients with MTHFR mutation and low folate levels presented the highest homocysteine levels. Conclusions— High plasma concentrations of homocysteine and low plasma folate levels were associated with an increased risk of CVT in this population in which low socioeconomic conditions and deficient nutritional status may contribute to its relatively high incidence.

The aim of this work was to study the microbiological and physico-chemical changes in four batches of Genestoso cheese (two ripened in autumn–winter and the other two in spring–summer) throughout manufacture and ripening. In general, the pH decreased more quickly in the cheeses manufactured in spring which, moreover showed values lower in aw and higher in salt/moisture content. The highest counts of aerobic mesophilic bacteria, presumptive lactococci and presumptive leuconostoc were reached in 2 day old cheeses. Lactobacilli and fungic flora became predominant with ripening. The counts of enterococci differed between the batches, but in general the maximum counts in kanamycin aesculin azide were obtained after ripening for 2 or 7 days. The counts on mannitol salt agar were very low and disappeared in the first days of ripening. Enterobacteriaceae were not detected after 15 days of ripening in the cheeses manufactured in spring nor in the 7 day old cheeses of an autumn batch.

Exposure to urban airborne particulate matter (PM) is associated with adverse health effects. We previously reported that the cytotoxic and proinflammatory effects of Mexico City PM10 (less than or equal to 10 micro m mean aerodynamic diameter) are determined by transition metals and endotoxins associated with these particles. However, PM2.5 (less than or equal to 2.5 micro m mean aerodynamic diameter) could be more important as a human health risk because this smaller PM has the potential to reach the distal lung after inhalation. In this study, we compared the cytotoxic and proinflammatory effects of Mexico City PM10 with those of PM2.5 using the murine monocytic J774A.1 cell line in vitro. PMs were collected from the northern zone or the southeastern zone of Mexico City. Elemental composition and bacterial endotoxin on PMs were measured. Tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) production by J774A.1 cells was measured in the presence or absence of recombinant endotoxin-neutralizing protein (rENP). Both northern and southeastern PMs contained endotoxin and a variety of transition metals. Southeastern PM10 contained the highest endotoxin levels, 2-fold higher than that in northern PM10. Northern and southeastern PM2.5 contained the lowest endotoxin levels. Accordingly, southeastern PM10 was the most potent in causing secretion of the proinflammatory cytokines TNF-alpha and IL-6. All PM2.5 and PM10 samples caused cytotoxicity, but northern PMs were the most toxic. Cytokine secretion induced by southeastern PM10 was reduced 50-75% by rENP. These results indicate major differences in PM10 and PM2.5. PM2.5 induces cytotoxicity in vitro through an endotoxin-independent mechanism that is likely mediated by transition metals. In contrast, PM10 with relatively high levels of endotoxin induces proinflammatory cytokine release via an endotoxin-dependent mechanism.

© 2015 American Chemical Society. In sparkling wines, mannoproteins released during yeast autolysis largely affect their final quality. This process is very slow and may take several months. The aim of this work was to study the effect of several commercial dry yeast autolysates on the chemical composition, foam, and sensory properties of white and rosé sparkling wines aged on lees for 9 months during two consecutive vintages. The addition of these products in the tirage phase did not affect either the content of phenolic compounds, amino acids, and biogenic amines or the foam properties. The commercial product with the highest mannoprotein content and the highest purity caused significant changes in the volatile composition of the wines and enhanced the fruity aromas in both Verdejo and Godello sparkling wines., We thank the Instituto Nacional de Investigación y Tecnologı́a Agraria y Alimentaria (INIA) for the funding provided for this study through Project RTA2009-029-C02 (with FEDER funds). L.M.-L. also thanks La Rioja Government for the financing of her predoctoral fellowship.

The aim of this work was focused on the study of the influence of grape variety and aging time in contact with lees and without lees, on volatile composition and foamability of white and rosé sparkling wines. Seven different grape varieties were used and the sparkling wines were studied until 30 months of aging on lees and after 12 months of aging in bottle after disgorging. Sparkling wines from Albarín, Verdejo, Godello and Prieto Picudo grape varieties were the richest in most of the volatile compounds analyzed, especially those that contribute to the fruity aroma of wines, and maintained their varietal characteristics even after long aging time (30 months). Verdejo and Prieto Picudo sparkling wines presented the best foam characteristics, followed by Albarín and Godello wines. Considering all the results, Albarín, Verdejo, Godello and Prieto Picudo were the most interesting grape varieties to elaborate sparkling wines, following the traditional or “champenoise” method., The authors would like to thank the Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria for the funding provided for this study through the project RTA2009-029-C02 (with FEDER funds). L. Martínez also thanks to La Rioja Government for the financing of her pre-doctoral fellowship.

© 2014 Elsevier Ltd. All rights reserved. The chemical composition of sparkling wines is directly related to their foam quality, but the compounds responsible are not yet completely established. This work aims at identifying the contribution of the different wine compounds to the foaming properties of white and rosé sparkling wines. Our results demonstrated the positive contribution of anthocyanins and amino acids to the foamability parameters HM (maximum height reached by foam after CO2 injection) and HS (foam stability height during CO2 injection), and the negative contribution of proanthocyanidins. Mannoproteins and polysaccharides rich in arabinose and galactose (PRAG) were poor foam formers but good foam stabilizers. The different forms of malvidin showed the highest influence on the HM and HS parameters, followed by amino acid compounds, mainly β-alanine. The model to explain foam stability was only predicted by polysaccharides from grapes, concretely PRAG. To our knowledge, this is the first time these correlations in sparkling wines have been described., The authors would like to thank the ‘Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria’ (INIA) for the funding provided for this study through the project RTA2009-029-C02 (with FEDER funds). Leticia Martínez-Lapuente also thanks La Rioja Government for the financing of her pre-doctoral fellowship.

The aim of this work was to study the microbiological, physico-chemical and proteolytic changes in Valdeon blue-veined cheese during ripening. Eight replicas of cheese were produced and a total of 48 cheeses were analysed. Lactic acid bacteria, mainly lactococci, were the predominant flora during the early stages of ripening, gradually being replaced by moulds and yeasts (8 log units). Enterococci and Enterobacteriaceae counts were very low or zero. This variety was characterised by a total solids content of 61.80g per 100g(-1) of cheese, a salt/moisture ratio of 8.92g salt per 100g(-1) moisture, a pH of 6.4-7.6 and a water activity of 0.917. At the end of ripening, primary and secondary proteolysis were very high, which resulted in an almost total degradation of αs1- and β-casein (approximately 90%). The peptide profile of the aqueous soluble extracts at pH 4.6 showed great complexity during ripening.

β-conglutin from Lupinus campestris was extracted and purified by gel filtration chromatography. The protein with molecular mass around 190 kDa showed by SDS-PAGE three major polypeptides of 53, 60 and 63 kDa and a minor polypeptide of 30 kDa. Isoelectric focusing of the native protein gave three isoforms with isoelectric points of 6.2, 6.3 and 6.4. Its sedimentation coefficient was 8.0S, with a hydrodynamic diameter of 110 A and possessed antigenic determinants in common with 7S proteins from common bean and soybean.

An aerobiological study was performed to evaluate the potential exposure of animals and workers to dust constituents generated during routine animal house work. Different rooms of air conditioned (A, control) and passively ventilated (B, non-air conditioned) animal facilities were sampled, in order to evaluate total airborne culturable fungi and bacteria, fungal spore concentrations and particle levels. Airborne room particles were analyzed gravimetrically and for endotoxin content. All parameters, except for culturable fungi, were higher in facility B and statistically significant, with respect to those from the control facility A. Median values for airborne particle concentration, endotoxin and fungal spores in facility B were: 115 µg m−3, 25 EU m−3, and 2173 spores m−3, respectively. Median values for facility A were: 66 µg m−3, 9 EU m−3, and 248 fungal spores m−3. Broncheoalveolar lavage from rats kept in the rat room of B, presented median concentrations of total cells and lactate dehydrogenase, higher than those found in the control facility (4.4 × 105vs. 1.1 × 105 and 2.7 UmL-1vs. 0.39 UmL−1, respectively). Values of total and biological particles of both facilities, as well as the time spent in different rooms, showed that worker exposure was higher during cage washing. It was especially high in the passively ventilated facility (airborne particles 686 µg m−3 3.5 h−1vs. 976 µg m−3 3.5 h−1, endotoxin 70 EU m−3 3.5 h−1vs. 108 EU m−3 3.5 h−1). The type of basidiospores and ascospores found, as well as the significant correlation between particle levels and endotoxin contents suggests that wood chip bedding disturbance during cage washing is an important source for airborne biological particles. The changes in broncheoalveolar lavage components found in rats from these facilities and previously reported changes in pro-inflammatory cellular responses found in workers, indicate that these relatively low levels of exposure are enough to induce a biological response. Studies considering the composition of mixed organic dusts, would be needed to reevaluate current occupational standards.

13 Págs., 3 Figs., 4 Tabls., Water-deficit irrigation to grapevines reduces plant growth, yield, and berry growth, altering the ripening process, all of which may influence fruit composition and wine quality. Therefore, the goals of this study were (1) to investigate the influence of the main endogenous berry hormones, abscisic acid (ABA), indole-3-acetic acid (IAA), salicylic acid (SA), and jasmonic acid (JA), on berry growth and ripening under water-deficit conditions and (2) to analyze changes in fruit composition, specifically N compounds, under water deprivation. The study was carried out using container-grown Tempranillo grapevines grown under controlled conditions in a greenhouse. Two irrigation treatments were imposed: control (well-watered) and sustained deficit irrigation (SDI). Water deficit decreased leaf area and the source-to-sink ratio, reduced yield and berry size, and decreased concentrations of the main phenolic compounds. SDI also modified berry hormonal status. At the pea-size stage, SDI berries had lower IAA and higher JA and SA than nonstressed berries. At veraison (onset of ripening), accumulation of ABA was less accentuated in SDI than in control berries. At harvest, the content of amino acids and free ammonium was low in both treatments but SDI-treated berries showed a significant accumulation of amines. Results suggest that water restrictions to grapevines might be playing a physiological role in reducing berry growth through affecting hormone dynamics, phenolic synthesis, and the berry amino acid content and composition, which could compromise fruit quality. Possible roles of endogenous IAA controlling berry size and endogenous ABA and SA controlling levels of anthocyanins and flavonols at harvest are discussed. © 2013 Springer Science+Business Media New York., This work was supported by Fundación Universitaria de Navarra (2011) and Ministerio de Ciencia e Innovación (MCINN BFU2011-26989), Spain. F. Morales thanks Gobierno de Aragón (A03 research group) for financial support. M. Niculcea was the recipient of a grant from Asociación de Amigos de la Universidad de Navarra.

The evolution in polysaccharide composition and molecular weights during sparkling wine making and aging was studied for the first time in this work. Different autochthonous grape varieties from Spain (Verdejo, Viura, Malvasía, Albarín, Godello, Garnacha and Prieto Picudo) were used to elaborate sparkling wines following the champenoise method. Principal component analysis showed differentiation of wines according to polysaccharide families. This differentiation was due to the process of aging on yeast lees, but not to the variety employed. The content of mannoproteins during aging was positively correlated (r = 0.792) with total polysaccharides from grapes. After six months of aging the highest content of mannoproteins and polysaccharides rich in arabinose and galactose was obtained. Also a shift to lower molecular weights was observed. The combination of these two characteristics could imply a better foam stability and thus sensory quality of sparkling wines. © 2013 American Chemical Society., The authors would like to thank the “Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria” (INIA) for the funding provided for this study through the project RTA2009-029-C02 (with FEDER funds). L. Martínez-Lapuente also thanks the La Rioja Government for the financing of her predoctoral fellowship.

This research studies the sensory properties and chemical composition of varietal wines made with minority varieties from La Rioja (Spain) and it analyses the transformations in phenolic compounds and colour parameters occurring during the malolactic fermentation. In this sense, all the analysed parameters underwent changes of the same magnitude during this stage and both anthocyanin and hydroxycinnamic acid distribution was found to be dependent on the grape variety and not on the winemaking process. Wines made with these varieties showed high values of resveratrol that could lead to healthier wines, and the variety Maturana Tinta de Navarrete was found to share quite similarities in the chemical parameters with Tempranillo, studied as a reference variety. In the sensory analysis, and although all the wines obtained good punctuations, Monastel and Maturana Tinta de Navarrete were the best valued. Monastel had the highest colour intensity and both varieties showed high aromatic intensity and great complexity. In mouth, Monastel showed the greater persistence, mount length and structure and Maturana Tinta de Navarrete was described as fresh and pleasant. In conclusion, this research shows the first characterization of these wines and provides data to be used as a chemotaxonomic tool to fingerprint them. Moreover, it opens the door to the use of minority varieties providing a viable alternative to traditional grape varieties cultivated in La Rioja and favouring consumer offer and wine differentiation. © 2012 Elsevier Ltd.

The mechanisms involved in sepsis-induced acute kidney injury (AKI) are unknown. We investigated the role of nitrosative stress in sepsis-induced AKI by studying the effects of manganese (III) tetrakis-(1-methyl-4-pyridyl) porphyrin pentachloride (MnTMPyP), a peroxynitrite decomposition catalyst, and aminoguanidine (AG), a selective nitric oxide synthase 2 (NOS2) inhibitor and peroxynitrite scavenger, on kidney function of rats subjected to cecal ligation and puncture (CLP). Sprague-Dawley rats (weighing 350 [SD, 50] g) were treated with MnTMPyP (6 mg/kg i.p.) or AG (50 mg/kg i.p.) at t = 12 and 24 h after CLP or sham procedure. At t = 36 h, mean arterial pressure and aortic blood flow were measured, and blood and urine samples were obtained for biochemical determinations, including creatinine clearance, fractional excretion of sodium, and neutrophil gelatinase-associated lipocalin concentration in the urine. Kidney tissue samples were obtained for (i) light microscopy, (ii) immunofluorescence and Western blot for 3-nitrotyrosine and NOS2, (iii) gene expression (quantitative real-time polymerase chain reaction) studies (NOS1, NOS2, NOS3, and superoxide dismutase 1), and (iv) matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Mean arterial pressure was unchanged and aortic blood flow decreased 25% in CLP animals. The sepsis-induced (i) decreased urine output and creatinine clearance and increased fractional excretion of sodium and urinary neutrophil gelatinase-associated lipocalin concentration, (ii) increased protein nitration and NOS2 protein, and (iii) NOS1 and NOS2 upregulation were all significantly attenuated by treatment with MnTMPyP or AG. Nitrated proteins in renal tissue from CLP animals (matrix-assisted laser desorption ionization time-of-flight mass spectrometry) were glutamate dehydrogenase, methylmalonate-semialdehyde dehydrogenase, and aldehyde dehydrogenase, mitochondrial proteins involved in energy metabolism or antioxidant defense. Nitro-oxidative stress is involved in sepsis-induced AKI, and protein nitration seems to be one mechanism involved.

The purpose of this study was to investigate how surfactant membranes can be perturbed by C-reactive protein (CRP) and whether surfactant protein A (SP-A) might overcome CRP-induced surfactant membrane alterations. The effect of CRP on surfactant surface adsorption was evaluated in vivo after intratracheal instillation of CRP into rat lungs. Insertion of CRP into surfactant membranes was investigated through monolayer techniques. The effect of CRP on membrane structure was studied through differential scanning calorimetry and fluorescence spectroscopy and microscopy using large and giant unilamellar vesicles. Our results indicate that CRP inserts into surfactant membranes and drastically increases membrane fluidity, resulting in surfactant inactivation. At 10% CRP/phospholipid weight ratio, CRP causes disappearance of liquid-ordered/liquid-disordered phase coexistence distinctive of surfactant membranes. SP-A, the most abundant surfactant lipoprotein structurally similar to C1q, binds to CRP (K(d)=56±8 nM), as determined by solid-phase binding assays and dynamic light scattering. This novel SP-A/CRP interaction reduces CRP insertion and blocks CRP effects on surfactant membranes. In addition, intratracheal coinstillation of SP-A+CRP into rat lungs prevents surfactant inhibition induced by CRP, indicating that SP-A/CRP interactions might be an important factor in vivo in controlling harmful CRP effects in the alveolus.

To demonstrate that increased formation of reactive oxygen (ROS) and nitrogen species (RNS) is involved in VILI-induced vascular dysfunction.Male Sprague-Dawley anesthetized rats were ventilated for 60 min using low V(T) ventilation [V(T) 9 ml/kg, positive end-expiratory pressure (PEEP) 5 cmH(2)O, n = 18], and high V(T) ventilation (V(T) 35 ml/kg, zero PEEP, n = 18). Arterial pressure and respiratory system mechanics were monitored. Blood samples for the determination of arterial blood gases and lactate concentration were drawn. Vascular rings from the thoracic aortae were mounted in organ baths for isometric tension recording. We studied endothelium-dependent relaxation in norepinephrine-precontracted rings (acetylcholine, 10 nM-10 microM) and contraction induced by norepinephrine (1 nM-10 microM) in resting vessels. Vascular rings were preincubated for 30 min with Zn-Mn-SOD (100 u/ml) or tempol (10(-4) M) (extracellular and intracellular superoxide scavengers, respectively) or MnTMPyP (10(-5) M) (a superoxide and peroxynitrite scavenger). The presence of superoxide and nitrotyrosine in aortic rings was evaluated by immunofluorescence.High V(T) ventilation induced hypotension, systemic acidosis, hypoxemia and hyperlactatemia, as well as impairment in acetylcholine and norepinephrine-induced responses in vitro. Responses to acetylcholine were improved by tempol (P = 0.004) and completely corrected (P0.001) by MnTMPyP. Responses to norepinephrine were also improved by treatment with tempol (P0.001) and MnTMPyP (P0.001). However, Zn-Mn-SOD did not improve acetylcholine- or norepinephrine-induced responses. Immunostaining for both superoxide and nitrotyrosine was increased in aortic rings from the high V(T) group.Our data support a role for intracellular ROS and peroxynitrite in the high V(T) ventilation-induced vascular dysfunction.

In the evaluation of particulate matter (PM) toxicity, one faces the challenge of identifying components that could be addressed as markers of toxicity. This study examines the use of statistical methods to determine which components present in the complex mixtures are related to toxic effects. The work is based on data previously published, where we demonstrated that particles collected in different zones of Mexico City showed different ability of inducing cell death, causing DNA damage, or creating proinflammatory effects. Empirically, we correlated those differences to variations in the concentration of transitional metals or endotoxin. In order to test those correlations, in this study we evaluated the role of the concentration of six transitional metals (Ni, V, Zn, Cu, Fe, and Pb) and endotoxin using the following tools: Pearson's regression analysis, correlation matrix between components, and multiple regression followed by a stepwise analysis. Due to collinearity found among several of the components, only Ni, Zn, and Pb were considered as independent variables. Among these, Ni and Zn show better correlations and help to explain the loss in viability, whereas Pb is the component that betters explains the proinflammatory effects. The statistical identification of composition markers facilitates the generation of a hypothesis relating the role played by the composition of PM and its biological effects.

Background and Purpose: Elevated homocysteine (Hcy) plasma levels are associated with an increased risk of spontaneous cervical artery dissection (sCAD). We examined the potential association between Hcy, folate, vitamin B12 levels and 5,10-methylenetetrahydrofolate reductase (MTHFR) polymorphisms in patients with cerebral infarct caused by sCAD. Patients and Methods: 39 patients who survived a cerebral infarct caused by sCAD [20 (51%) women; 24 (61.5%) vertebral and 15 (38.5%) internal carotid arteries], and 76 healthy control subjects were included. Hcy plasma levels (fasting and after methionine load), folate and vitamin B12 levels were measured. We also performed polymorphisms of MTHFR. Hcy, vitamin B12, folates and polymorphisms of MTHFR were assessed and any associations were analyzed using multivariate statistics. Results: Mean plasma fasting Hcy level was 9.81 µmol/l for cases and 6.38 for controls (p = 0.001). The occurrence of sCAD was associated with elevated fasting Hcy levels (>95th percentile over the control group) with an adjusted odds ratio of 7.9 (95% CI 1.66–35). The association between low plasma folate values (MTHFR genotype showed a higher TT mutant frequency among CAD patients (p = 0.034). Conclusions: High plasma concentrations of Hcy and low plasma levels of folate were associated with an increased risk of sCAD in the sample studied. We conclude that deficiencies in nutritional status may contribute to the relatively high incidence of CAD in Mexico.