A highactivity ATP translocator in mesophyll chloroplasts of Digitaria sanguinalis, a plant having the C-4 dicarboxylic acid pathway of photosynthesis

The effect of exogenous adenine nucleotides on CO2 fixation and oxygen evolution was studied with mesophyll protoplast extracts of the C4 plant Digitaria sanguinalis. Exogenous ATP was found to stimulate the rate of pyruvate and pyruvate+oxalacetate induced CO2 fixation, as well as reverse the inhibition of CO2 fixation of carbonyl cyanide m-chlorophenyl hydrazone and several electron transport inhibitors. The ATP-dependent stimulation of CO2 fixation varied from 40 to 70 mumol CO2 fixed/mg chlorophyll per h, suggesting that ATP was crossing the chloroplast membranes at rates of 80-140 mumol/mg chlorophyll per h, since 2 ATP are required for each CO2 fixed. Fixation of CO2 could also be induced in the dark by exogenous ATP, in which case ADP accumulated outside the chloroplasts...     

Molecular studies to identify the Fusarium species responsible for HT-2 and T-2 mycotoxins in UK oats

High levels of Fusarium mycotoxins HT-2 and T-2 have been detected in UK oats since surveys started in 2002. Fusarium langsethiae and the closely related species F. sporotrichioides have previously been associated with the contamination of cereals with type A trichothecenes HT-2 and T-2 in Nordic countries. Preliminary microbiological analysis of UK oat samples with high concentrations of HT-2 and T-2 detected and isolated F. langsethiae and F. poae but not the other type A trichothecene producing species F. sporotrichioides, F. sibiricum and F. armeniacum. Two hundred and forty oat flour samples with a known mycotoxin profile were selected from a previous four year study (2002-2005) to cover the full concentration range from below the limit of quantification (<20 μg/kg) to 9,990 μg/kg HT-2+T-2 combined. All samples were analysed for the DNA of F. langsethiae, F. poae and F. sporotrichioides based on previously published PCR assays. F. langsethiae was detectable in nearly all samples; F. poae was detected in 90% of samples whereas F. sporotrichioides was not detected in any sample. A real-time PCR assay was developed to quantify F. langsethiae DNA in plant material. The assay could quantify as low as 10(-4)ngF. langsethiae DNA/μl. Based on this assay and a previously published assay for F. poae, both species were quantified in the oat flour samples with known HT-2+T-2 content. Results showed a good regression (P<0.001, r(2)=0.60) between F. langsethiae DNA and HT-2+T-2 concentration. F. poae DNA concentration was not correlated to HT2+T2 concentration (P=0.448) but was weakly correlated to nivalenol concentration (P<0.001, r(2)=0.09). Multiple regression with F. langsethiae and F. poae DNA as explanatory variates identified that both F. langsethiae and F. poae DNA were highly significant (P<0.001) but F. poae DNA only accounted for an additional 4% of the variance and the estimate was negative, indicating that higher concentrations of F. poae DNA were correlated with slightly lower concentrations of HT2+T2 detected. A stronger regression (P<0.001, r(2)=0.77) between F. langsethiae DNA and HT-2+T-2 was obtained after extraction and quantification of DNA and mycotoxins from individual oat grains. The results from this study provide strong evidence that F. langsethiae is the primary, if not sole, fungus responsible for high HT-2 and T-2 in UK oats.     

Gene expression profiles for detecting and distinguishing potential endocrine-disrupting compounds in environmental samples

Industrial and municipal processes may produce and release endocrine-disrupting compounds (EDCs) into the environment, but the exact nature of their effects is difficult to investigate. EDCs typically exert their effect by affecting gene expression aberrantly. To determine if gene expression profiles could be used to detect and distinguish estrogenic EDCs, an estrogen receptor positive human breast cancer cell line (MCF-7) was exposed to known estrogenic compounds, suspected EDCs, and extracts from three effluent samples. A set of specifically estrogen-regulated genes was identified by microarray analysis. Nine estrogen up-regulated genes (IGFBP4, HSPA8, B4GALT1, XBP1, KRT8, GTPBP4, HNRPAB, SLC2A1, and CALM1) and two estrogen down-regulated genes (ID2 and ZNF217) were consistently detectable in response to estrogen and other estrogenic compounds. Gene expression patterns in cells that were exposed to effluent sample extracts were compared to gene expression patterns in cells that were exposed to known endocrines. Using this technique, two of the effluent samples were shown to have estrogenic activity. This approach could easily be extended to screen for other types of receptor-mediated endocrine disruption. For example, cells expressing androgen or aryl hydrocarbon receptors could be used in gene expression profiling assays to detect androgenic effects or for the presence of bioactive aromatic hydrocarbons. Gene expression profiling is emerging as a sensitive and specific method to screen complex samples for endocrine disrupting activity.     

Using activity and milk yield as predictors of fresh cow disorders

The objective was to determine whether daily walking activity and milk yields could be used as predictors of metabolic and digestive disorders early in lactation. Data were collected from 1996 through 1999 from 1445 dairy cows in 3 Florida herds. Walking activity, milk yield, and other measures were collected from a computerized dairy management system. Mixed models analysis was used for data on cows before their first detected estrus, as identified by difference in activity. Healthy cows were defined as those without any metabolic or digestive disorder during the prebreeding stage, whereas a sick cow had an occurrence of those disorders at any time during the prebreeding stage. Metabolic disorders were ketosis, retained placenta, and milk fever. Digestive disorders included displaced abomasum, indigestion, reduced feed intake, traumatic gastritis, acidosis, and bloat. Data from cows with known cases of ketosis, left displaced abomasum, and digestive disorders were analyzed to determine changes in activity and milk yield before those specific disorders were clinically diagnosed. Although walking activity was generally lower among sick cows, cows with ketosis, left displaced abomasum, and digestive disorders had higher than average activity 8, 9, and 8 d, respectively, before each diagnosed disorder. Daily milk yields of sick cows were approximately 15 kg/d less than milk yields of healthy cows. Milk yields were lower by 6, 7, and 5 d, respectively, before diagnoses of ketosis, left displaced abomasum, and digestive disorders. Cows with ketosis, left displaced abomasum, and general digestive disorders could possibly be detected about 5 to 6 d earlier than clinical diagnoses based on changes in daily walking activity and milk yield.     

Fluorotelomer alcohol biodegradation yields poly- and perfluorinated acids

The widespread detection of environmentally persistent perfluorinated acids (PFCAs) such as perfluorooctanoic acid (PFOA) and its longer chained homologues (C9>C15) in biota has instigated a need to identify potential sources. It has recently been suggested that fluorinated telomer alcohols (FTOHs) are probable precursor compounds that may undergo transformation reactions in the environment leading to the formation of these potentially toxic and bioaccumulative PFCAs. This study examined the aerobic biodegradation of the 8:2 telomer alcohol (8:2 FTOH, CF3(CF2)7CH2CH2OH) using a mixed microbial system. The initial measured half-life of the 8:2 FTOH was approximately 0.2 days mg(-1) of initial biomass protein. The degradation of the telomer alcohol was monitored using a gas chromatograph equipped with an electron capture detector (GC/ECD). Volatile metabolites were identified using gas chromatography/ mass spectrometry (GC/MS), and nonvolatile metabolites were identified and quantified using liquid chromatography/ tandem mass spectrometry (LC/MS/MS). Telomer acids (CF3(CF2)7CH2COOH; CF3(CF2)6CFCHCOOH) and PFOA were identified as metabolites during the degradation, the unsaturated telomer acid being the predominant metabolite measured. The overall mechanism involves the oxidation of the 8:2 FTOH to the telomer acid via the transient telomer aldehyde. The telomer acid via a beta-oxidation mechanism was furthertransformed, leading to the unsaturated acid and ultimately producing the highly stable PFOA. Telomer alcohols were demonstrated to be potential sources of PFCAs as a consequence of biotic degradation. Biological transformation may be a major degradation pathway for fluorinated telomer alcohols in aquatic systems.     

Variability in carbon isotopic fractionation during biodegradation of chlorinated ethenes: implications for field applications

Stable carbon isotopic analysis has the potential to assess biodegradation of chlorinated ethenes. Significant isotopic shifts, which can be described by Rayleigh enrichment factors, have been observed for the biodegradation of trichloroethlyene (TCE), cis-dichloroethylene (cDCE), and vinyl chloride (VC). However, until this time, no systematic investigation of isotopic fractionation during perchloroethylene (PCE) degradation has been undertaken. In addition, there has been no comparison of isotopic fractionation by different microbial consortia, nor has there been a comparison of isotopic fractionation by consortia generated from the same source, but growing under different conditions. This study characterized carbon isotopic fractionation during reductive dechlorination of the chlorinated ethenes, PCE in particular, for microbial consortia from two different sources growing under different environmental conditions in order to assess the extent to which different microbial consortia result in different fractionation factors. Rayleigh enrichment factors of -13.8@1000, -20.4@1000, and -22.4@1000 were observed for TCE, cDCE, and VC, respectively, for dechlorination by the KB-1 consortium. In contrast, isotopic fractionation during reductive dechlorination of perchloroethylene (PCE) could not always be approximated by a Rayleigh model. Dechlorination by one consortium followed Rayleigh behavior (epsilon = -5.2), while a systematic change in the enrichment factor was observed over the course of PCE degradation by two other consortia. Comparison of all reported enrichment factors for reductive dechlorination of the chlorinated ethenes shows significant variation between experiments. Despite this variability, these results demonstrate that carbon isotopic analysis can provide qualitative evidence of the occurrence and relative extent of microbial reductive dechlorination of the chlorinated ethenes.     

Stormwater and fire as sources of black carbon nanoparticles to Lake Tahoe

Emitted to the atmosphere through fire and fossil fuel combustion, refractory black carbon nanoparticles (rBC) impact human health, climate, and the carbon cycle. Eventually these particles enter aquatic environments, where they may affect the fate of other pollutants. While ubiquitous, the particles are still poorly characterized in freshwater systems. Here we present the results of a study determining rBC in waters of the Lake Tahoe watershed in the western United States from 2007 to 2009. The study period spanned a large fire within the Tahoe basin, seasonal snowmelt, and a number of storm events, which resulted in pulses of urban runoff into the lake with rBC concentrations up to 4 orders of magnitude higher than midlake concentrations. The results show that rBC pulses from both the fire and urban runoff were rapidly attenuated suggesting unexpected aggregation or degradation of the particles. We find that those processes prevent rBC concentrations from building up in the clear and oligotrophic Lake Tahoe. This rapid removal of rBC soon after entry into the lake has implications for the transport of rBC in the global aquatic environment and the flux of rBC from continents to the global ocean.     

NMR-based metabonomics detection of differences in the metabolism of hydrolysed versus intact protein of similar amino acid profile

BACKGROUND: Proton nuclear magnetic resonance (NMR)‐based metabonomics has only recently been applied to nutritional research. The limitation of any analytical technique is its sensitivity in detecting the smallest variation. Alterations in nutrition often produce only subtle metabolic modulations. The objective of this study was to determine if NMR‐based metabonomics could detect variations in the metabolic profile of urine from pigs digesting either native casein (NC) or the same casein that had been enzymatically hydrolysed (EHC). NMR permits simultaneous detection of a large number of metabolites, thus allowing detection of unanticipated metabolic fluctuations that may otherwise have gone undetected with the use of only targeted analysis. RESULTS: Partial least squares discriminant analysis identified significantly (P < 0.05) higher urinary excretions of leucine, valine, taurine and glycine by pigs on the EHC‐based diet. CONCLUSION: NMR‐based metabonomics is a sensitive method that can uncover unanticipated metabolic changes brought about by physicochemical changes to the feedstock (i.e. hydrolysis). The data show a lower efficiency of retention by the kidney of some amino acids following ingestion of a hydrolysed protein. Copyright © 2012 Society of Chemical Industry     

Meat quality in pigs subjected to minimal preslaughter stress

The experiment compared meat quality in carcasses from pigs subjected to the absolute minimum of stress preslaughter with that from carcasses of pigs subjected to simulated commercial handling. A total of 120 pigs (85 kg) were used. They were a commercial hybrid and were processed in six equal batches. The minimal stress group were slaughtered with as little stress as possible. The simulated commercial handling group were mixed and transported for 2hr (80 km) and lairaged for 30 min. Simulated commercial handling increased plasma cortisol, plasma lactate, CPK and beta-endorphin. Muscle temperature measured in the m. longissimus dorsi (LD) at 45 min increased as did the ultimate pH in the m. adductor (AD). Overall eating quality was not affected. There were batch affects on most variables. Males were leaner and produced meat that had greater abnormal fat odour, darker cooked colour, poorer pork flavour, lower overall acceptability and was tougher.