Bioreduction of trichloroethene using a hydrogen-based membrane biofilm reactor

A H2-based, denitrifying membrane-biofilm reactor (MBfR) was effective for removing trichloroethene (TCE) by reductive dechlorination. When TCE was first added to the MBfR, reductive dechlorination took place immediately and then increased over 18 weeks, and TCE was completely dechlorinated to ethene by about 120 days. These results indicate that TCE-dechlorinating bacteria were present naturally in the H2-based biofilm, and that enrichment for TCE-dechlorinating bacteria occurred. Dehalococcoides were documented in the MBfR biofilm before and after TCE feeding. Their proportion, quantified using the 16S rRNA gene, increased from 2.9 to 12% after TCE addition. This is the first report in which Dehalococcoides are proven to be present as part of an autotrophic biofilm community active in reductive dechlorination of TCE to ethene in a laboratory controlled experiment. Based on the complete reduction of TCE to ethene, the 16S rRNA clone libraries results, and the amount of tceA and bvcA, it appears that at least two Dehalococcoides strains were present in the enriched biofilm. One of them seems to be a new strain that is unique for having tceA and bvcA reductive dehalogenases.     

Grain challenge affects systemic and hepatic molecular biomarkers of inflammation,stress, and metabolic responses to a greater extent in Holstein than Jersey cows

Long-term feeding of high-grain diets to dairy cows often results in systemic inflammation characterized by alterations in acute-phase proteins and other biomarkers, both in plasma and immune-responsive tissues like the liver. The molecular and systemic changes that characterize an acute grain feeding challenge remain unclear. The current study involved 6 Holstein and 6 Jersey cows in a replicated 2 × 2 Latin square. Periods (10 d) were divided into 4 stages (S): S1, d 1 to 3, served as baseline with total mixed ration (TMR) ad libitum; S2, d 4, served as restricted feeding, with cows offered 50% of the average daily intake observed in S1; S3, d 5, a grain challenge was performed, in which cows were fed a TMR ad libitum without (CON) or with an additional pellet wheat-barley (1:1; HIG) at 20% of dry matter intake top-dressed onto the TMR; S4, d 6 to 10, served as recovery during which cows were allowed ad libitum access to the TMR. Among the 28 biomarkers analyzed in blood 12 h after grain challenge on d 5, the concentrations of fatty acids and bilirubin increased in HIG Holstein but not Jersey cows. In Holsteins, feeding HIG also increased total protein and albumin while decreasing ceruloplasmin, myeloperoxidase, and alkaline phosphatase concentrations. At the molecular level, hepatic genes associated with inflammation (IL1B, IL6, TNF, TLR4, MYD88, and NFKB1) were upregulated in Holstein cows fed HIG versus CON. Despite such response, expression of the acute-phase proteins SAA and HP in Holsteins fed HIG compared with CON was markedly downregulated. In Holsteins fed HIG versus CON, the marked downregulation of SCD, ELOVL6, and MTTP along with upregulated CPT1A, ACOX1, and APOA5 indicated alterations in fatty acid and lipoprotein metabolism during grain challenge. Genes related to ketogenesis (HMGCS2 and ACAT1) were upregulated in Jerseys, and gluconeogenic genes (PDK4 and PCK1) were upregulated in Holstein cows fed HIG, suggesting alterations in ketone body and glucose production. Expression of phosphorylated p70S6K1, RPS6, and 4EBP1 proteins, as well as total mechanistic target of rapamycin (mTOR) protein, decreased in Holsteins fed HIG, whereas phosphorylated mTOR and 4EBP1 proteins increased in Jerseys fed HIG. From a metabolic and inflammatory biomarker standpoint, data indicate that Jersey cows better tolerated the acute grain challenge. Alterations in mTOR signaling proteins in both Jerseys and Holsteins fed HIG suggest a potential role for exogenous AA in the hepatic adaptations to grain challenge. It remains to be determined if these acute responses to a grain challenge can elicit long-term liver dysfunction, which could negatively affect welfare of the cow.     

The importance of landscape in wine quality perception: An integrated approach using choice-based conjoint analysis and combination-based permutation tests

This work’s aim is to evaluate the importance of landscape in wine quality perception using a novel integrated approach based on two statistical techniques, i.e. choice-based conjoint analysis (Gustafsson et al., 2001) and combination-based permutation tests (Corain and Salmaso, 2004, Pesarin and Salmaso, 2009a). To this end we developed a preference choice experiment which was submitted to a sample of respondents during blind wine tasting meetings. The obtained results allow us to weigh up and emphasise the importance of an attractive landscape to the perception of a finer quality of wine. Indeed, we noticed that associating wine to an image of greater visual impact, i.e. an evocative landscape, induces a significantly higher preference for the tasted wine. Moreover, the integrated application of choice-based conjoint analysis and combination-based permutation tests allows us to identify the specific relationship each market segment has with landscape, price and wine quality.     

Effect of natural fermentation on the content of inositol phosphates in lentils

The effects of natural fermentation upon phytic acid and less phosphorylated inositol phosphates ofLens culinaris var vulgaris cultivar Magda-20 were investigated. Seven fermentation runs were made following a 2² complete factorial design with three replicated centre points to study the effect of different conditions of temperature (28, 35 and 42°C) and broth concentration (79, 150 and 221 g/l). Samples were taken for each of them at daily intervals (0, 24, 48, 72 and 96 h). The pH value declined sharply in the first 24 h of fermentation, becoming stabilized from this time. The relation between lactic acid and titratable acidity presented important differences between the different fermentations, ranging from 30–80%. Phytic acid (IP ₆), inositol pentakis (IP ₅), tetrakis (IP ₄) and tris-(IP ₃) phosphates were quantitatively determined. The content of total inositol phosphates showed a maximum reduction of 63% at 72 h under the fermentation conditions of 42°C and 79 g/l.     

Association of hemoglobin Saint Etienne (alpha2beta295F8 His replaced by G1n) with hemoglobins A and F. Synthesis and subunit exchange in vitro

The unstable hemoglobin (Hb) Saint Etienne (alpha2beta295F8 His replaced by G1n) (betaSE) was found in the red blood cells of an 8-year-old boy. The composition of this hemoglobin was 26% Saint Etienne, 52% A, 3% A2 and 19% HbF. Studies of hemoglobin synthesis indicate: a) a balanced synthesis of alpha and non-alpha chains (alpha=betaA + betaSE + gamma), b) an increased pool of free alpha hemoglobin chains, and c) a rapid exchange of alpha chains between this pool and HbSE. The alpha chain pool resulted from the dissociation of HbSE and the greater instability of betaSE chains than alpha chains upon heating. Hemoglobin F is of the fetal type and is heterogeneously distributed among the red cells. Furthermore, two populations of red blood cells could be separated according to their i antigen content. Analysis of the hemoglobins revealed a heterogeneous distribution. Thus, F hemoglobin was preferentially associated with cells having low i antigen level, while Saint Etienne hemoglobin was increased in cells having a high i antigen level. HbF and HbSE were not present in the parents of the propositus. Study of the genetic markers confirmed the filiation. The parents were normal upon clinical and hematological examination; they exhibited a normal pattern and synthesis of hemoglobin. The Hb Saint Etienne case is compared with Hb Istanbul, which in spite of the same amino acid substitution is not associated with increased HbF level.     

Identification of Sialylated Oligosaccharides Derived from Ovine and Caprine Caseinomacropeptide by Graphitized Carbon Liquid Chromatography–Electrospray Ionization Ion Trap Tandem Mass Spectrometry

Isolation and characterization of oligosaccharides from caseinomacropeptide (CMP) are important in understanding the biological and functional properties of CMP. However, it is difficult to achieve this goal, due to the high degree of isomerism present in these types of compounds. In this study, the sialylated oligosaccharides derived from ovine and caprine CMP were released as oligosaccharide alditols by reductive β-elimination and subsequently separated and characterized using graphite carbon column liquid chromatography–negative electrospray ionization ion trap tandem mass spectrometry (LC/ESI(?)-MSⁿ). Although, the chromatographic resolution of isomeric oligosaccharides was not achieved perfectly, the characteristic tandem mass spectra of these compounds allowed differentiating and confirming unequivocally the structure of each one of the oligosaccharides. In CMP of both species, four trisaccharides and four tetrasaccharides were identified as O-glycans. Their chemical structures were identified to be Galβ1-3(NeuAcα2-6)GalNAcol, NeuAcα2-3Galβ1-3GalNAcol, Galβ1-3(NeuGcα2-6)GalNAcol, NeuGcα2-3Galβ1-3GalNAcol, NeuAcα2-3Galβ1-3(NeuGcα2-6)GalNAcol, NeuGcα2-3Galβ1-3(NeuAcα2-6)GalNAcol, NeuAcα2-3Galβ1-3(NeuAcα2-6)GalNAcol, and NeuGcα2-3Galβ1-3(NeuGcα2-6)GalNAcol. The LC/MSⁿ methodology using an ion trap-type mass analyzer shown in this study is of general applicability for determination of short O-glycan oligosaccharides.     

Radical Scavenging,Total Antioxidant Capacity,and Antiproliferative Activity of Phenolic Extracts from Extra Virgin Olive Oil by Cultivar ‘Frantoio’

The beneficial health effects of extra virgin olive oil are due to both its high content of monounsaturated fatty acids and its high content of phenolic compounds, which have recently attracted research interest. In this context, the aim of this work was to examine the in vitro antioxidant and antiproliferative activities of the phenolic extract obtained from extra virgin olive oil from O. europea cultivar ‘Frantoio’ (samples 1–4), one of the main varieties cultivated in Italy. The total phenolic content was determined by Folin–Ciocalteu method and instead the phenolic profile was obtained by HPLC coupled to a diode array detector and mass spectrometry. Extra virgin olive oil extracts exhibited high antioxidant activity through different mechanisms of action and these activities are related to the phenolic content. Sample 3 demonstrated the strongest free radical scavenging activity with IC₅₀ value of 56.5 μg/mL. The reducing ability measured with FRAP assay revealed that samples ranged from 91.3 to 156 μM Fe(II)/g. The same interesting trend was observed with Trolox equivalent antioxidant capacity value. Moreover, the virgin olive oils showed a good oxidative stability ranging between 19 to 32 h. Antiproliferative activity evaluated by SRB assay revealed that phenolic extracts from the cultivar ‘Frantoio’ showed a strong antiproliferative activity against CORL-23 cell line with an IC₅₀ value of 14.5 and 55.9 μg/mL for samples 3 and 1, respectively, and these results are comparable to the positive control vinblastine. Overall, these results showed that extra virgin olive oils from the cultivar ‘Frantoio’, may represent an interesting source of phenolic compounds as functional components that could be consumed in diets and/or used for the elaboration of functional food and pharmaceutical industries.     

Genetic analysis of leukosis incidence in United States Holstein and Jersey populations

Bovine leukosis (BL) is a retroviral disease caused by the bovine leukosis virus that affects only cattle. It is associated with decreased milk production and increased cull rates due to development of lymphosarcoma. The virus also affects the immune system. Infected cows display a weak response to some vaccinations. It is important to determine if the heritability of BL susceptibility is greater than zero, or if the environment is the only factor that can be used to reduce the transmission and incidence of the disease. Accordingly, the aim of this study was to estimate the heritability for BL incidence and the genetic merit of sires for leukosis resistance in Holstein and Jersey cattle. Continuous scores and binary milk ELISA results for 13,217 Holstein cows from 114 dairy herds across 16 states and 642 Jersey cows from 8 dairy herds were considered. Data were obtained from commercial testing records at Antel BioSystems (Lansing, MI). Out of the 13,859 animals tested, 38% were found to be infected with the disease. Linear and threshold animal models were used to analyze the continuous and binary data, respectively. Results from both models were similar in terms of estimated breeding values and variance components in their respective scales. Estimates of heritability obtained with the 2 approaches were approximately 8% for both breeds, indicating a considerable genetic component underlying BL disease incidence. The correlation between the estimated breeding values from the 2 models was larger than 0.90, and the lists of top 10% bulls selected from each model had about 80% overlap for both breeds. In summary, results indicate that a simple linear model using the continuous ELISA scores as the response variable was a reasonable approach for the genetic analysis of BL incidence in cattle. In addition, the levels of heritability found indicate that genetic selection could also be used to decrease susceptibility to bovine leukosis virus infection in Holstein and Jersey cattle. Further research is necessary to investigate the genetic correlations of BL with other production and reproduction traits, and to search for potential genomic regions harboring major genes affecting BL susceptibility.     

Identification and quantification of leaf surface flavonoids in wild-growing populations of Dracocephalum kotschyi by LC–DAD–ESI-MS

Dracocephalum kotschyi Boiss. (Lamiaceae) is an aromatic and perennial herb endemic to Iran with interesting pharmacological and biological properties. The flavonoids luteolin-7-O-glucoside, apigenin-7-O-glucoside (cosmosiin), luteolin 3′-O-β-d-glucuronide, luteolin, apigenin, cirsimaritin, isokaempferide, penduletin, xanthomicrol, calycopterin and the polyphenol rosmarinic acid were identified among 13 natural populations of the plant by ESI–MS, LC–DAD and LC–DAD–ESI-MS. The plant extracts containing the identified compounds showed significant antioxidant activity, which was correlated with the flavonoid content. Additionally, leaf and stem size and geographical variability among the studied populations were correlated with flavonoid accumulation. Canonical correlation analysis was used to find a relationship between plant dimensions and phytochemical composition, and the plants with the lowest growth indices were found to have the highest levels of methoxylated flavonoids.