The mobilization of copper in sheep by chelating agents.

D-penicillamine and 2,3-dimercapto-propane-l-sulphonate (DMPS) were administered orally and disodium-calcium-ethylene-diamine-tetraacetate (Ca-EDTA) subcutaneously into copper loaded sheep. The results showed that D-penicillamine has a copper mobilizing effect, while DMPS and Ca-EDTA do not seem to have this effect to any significant degree. Penicillamine, 52 mg/kg of body weight daily for 6 days, increased the urinary excretion of copper by a factor of 10 to 20. The same dosage (on weight basis) of DMPS increased the excretion of copper in urine by 2 fold. Following Ca-EDTA treatment no increase in urinary excretion of copper was observed. Penicillamine could be used in prophylactic treatment of copper toxicosis in sheep, but it is still too expensive for practical use.     

Enzymatic hydrogenation of trans-2-nonenal in barley

Conversion of undesirable, taste-active compounds is crucial for using barley as a suitable raw material for beer production. Here, ALH1, a barley alkenal hydrogenase enzyme that reduced the alpha,beta-unsaturated double bond of aldehydes and ketones, was found to convert trans-2-nonenal (T2N), a major contributor to the cardboard-like flavor of aged beer. Although the physiological function of ALH1 in barley development remains elusive, it exhibited high specificity with NADPH as a cofactor in the conversion of several oxylipins-including T2N, trans-2-hexenal, traumatin, and 1-octen-3-one. ALH1 action represents a previously unknown mechanism for T2N conversion in barley. Additional experimental results resolved the genomic sequence for barley ALH1, as well as the identification of a paralog gene encoding ALH2. Interestingly, T2N was not converted by purified, recombinant ALH2. The possibility to enhance ALH1 activity in planta is discussed--not only with respect to the physiological consequences thereof--but also in relation to improved beer quality.     

Two-year toxicity and carcinogenicity study of methyleugenol in F344/N rats and B6C3F(1) mice

Methyleugenol (MEG) was tested for toxicity/carcinogenicity in a 2-yr carcinogenesis bioassay because of its widespread use in a variety of foods, beverages, and cosmetics as well as its structural resemblance to the known carcinogen safrole. F344/N rats and B6C3F(1) mice (50 animals/sex/dose group) were given MEG suspended in 0.5% methylcellulose by gavage at doses of 37, 75, or 150 mg/kg/day for 2 yr. Control groups (60 rats/sex and 50 mice/sex) received only the vehicle. A stop-exposure group of 60 rats/sex received 300 mg/kg/day by gavage for 53 weeks followed by the vehicle only for the remaining 52 weeks of the study. A special study group (10 animals/sex/species/dose group) were used for toxicokinetic studies. All male rats given 150 and 300 mg/kg/day died before the end of the study; survival of female rats given 150 mg/kg/day and all treated female mice was decreased. Mean body weights of treated male and female rats and mice were decreased when compared to control. Area under the curve results indicated that greater than dose proportional increases in plasma MEG occurred for male 150 and 300 mg/kg/day group rats (6 and 12 month) and male 150 mg/kg/day mice (12 month). Target organs included the liver, glandular stomach, forestomach (female rats) and kidney, mammary gland, and subcutaneous tissue (male rats). Liver neoplasms occurred in all dose groups of rats and mice and included hepatoadenoma, hepatocarcinoma, hepatocholangioma (rats only), hepatocholangiocarcinoma, and hepatoblastoma (mice only). Nonneoplastic liver lesions included eosinophilic and mixed cell foci (rats only), hypertrophy, oval cell hyperplasia, cystic degeneration (rats only), and bile duct hyperplasia. Mice also exhibited necrosis, hematopoietic cell proliferation, and hemosiderin pigmentation. Glandular stomach lesions in rats and mice included benign and malignant neuroendocrine tumors, neuroendocrine cell hyperplasia, and atrophy and in mice included glandular ectasia/chronic active inflammation. In female rats, the forestomach showed a positive trend in the incidences of squamous cell papilloma or carcinoma (combined). Male rats also exhibited kidney (renal tubule hyperplasia, nephropathy, and adenomacarcinoma), mammary gland (fibroadenoma), and subcutaneous tissue (fibroma, fibrosarcoma) lesions. Male rats also exhibited malignant mesotheliomas and splenic fibrosis. These data demonstrate that MEG is a multisite, multispecies carcinogen.     

Combining conservative and variable markers to infer the evolutionary history of Prunus subgen. Amygdalus s.l. under domestication

The genus Prunus L. is large and economically important. However, phylogenetic relationships within Prunus at low taxonomic level, particularly in the subgenus Amygdalus L. s.l., remain poorly investigated. This paper attempts to document the evolutionary history of Amygdalus s.l. and establishes a temporal framework, by assembling molecular data from conservative and variable molecular markers. The nuclear s6pdh gene in combination with the plastid trnSG spacer are analyzed with bayesian and maximum likelihood methods. Since previous phylogenetic analysis with these markers lacked resolution, we additionally analyzed 13 nuclear SSR loci with the δµ² distance, followed by an unweighted pair group method using arithmetic averages algorithm. Our phylogenetic analysis with both sequence and SSR loci confirms the split between sections Amygdalus and Persica, comprising almonds and peaches, respectively. This result is in agreement with biogeographic data showing that each of the two sections is naturally distributed on each side of the Central Asian Massif chain. Using coalescent based estimations, divergence times between the two sections strongly varied when considering sequence data only or combined with SSR. The sequence-only based estimate (5 million years ago) was congruent with the Central Asian Massif orogeny and subsequent climate change. Given the low level of differentiation within the two sections using both marker types, the utility of combining microsatellites and data sequences to address phylogenetic relationships at low taxonomic level within Amygdalus is discussed. The recent evolutionary histories of almond and peach are discussed in view of the domestication processes that arose in these two phenotypically-diverging gene pools: almonds and peaches were domesticated from the Amygdalus s.s. and Persica sections, respectively. Such economically important crops may serve as good model to study divergent domestication process in close genetic pool.     

Heavy metals in tropical Lake Kariba,Zimbabwe

Metal mining is carried out in the drainage basin of Lake Kariba, Zimbabwe. In an attempt to assess the distribution of heavy metals in the lake ecosystem, the concentrations of chromium (Cr), cadmium (Cd), copper (Cu), lead (Pb), manganese (Mn), nickel (Ni), zinc (Zn), and selenium (Se) were analysed in the following ecosystem components at different locations in the lake; water, sediment, aquatic macrophytes, mussels, and carnivorous and herbivorous fish. Concentrations were generally in the same range as in other temperate and tropical aquatic ecosystems, except for Pb and Cd, which were higher in marcrophytes and fish from the lake. Apart from Cd, there were no signs of biomagnification and the highest levels of metals were found in the macrophytes and sediments. While metal contamination in fish is probably a combination of uptake from the food and an equilibrium with the surrounding water, the levels in macrophytes appear to be determined mainly by uptake from the sediment through the roots.     

Molecular cloning of the chicken progesterone receptor

To define the functional domains of the progesterone receptor required for gene regulation, complementary DNA (cDNA) clones encoding the chicken progesterone receptor have been isolated from a chicken oviduct lambda gt11 cDNA expression library. Positive clones expressed antigenic determinants that cross-reacted with six monospecific antibodies derived from two independent sources. A 36-amino acid peptide sequence obtained by microsequencing of purified progesterone receptor was encoded by nucleotide sequences in the longest cDNA clone. Analysis of the amino acid sequence of the progesterone receptor deduced from the cDNA clones revealed a cysteine-rich region that was homologous to a region found in the estrogen and glucocorticoid receptors and to the avian erythroblastosis virus gag-erb-A fusion protein. Northern blot analysis with chicken progesterone receptor cDNA's indicated the existence of at least three messenger RNA species. These messages were found only in oviduct and could be induced by estrogens.     

Kettle holes reflect the biogeochemical characteristics of their catchment area and the intensity of the element-specific input

Journal of Soils and Sediments - Kettle holes are small inland water bodies known to be dominated by terrigenous material; however, the processes and structures that drive the enrichment and...     

Co-incubation with IL-18 potentiates antigen-specific IFN-γ response in a whole-blood stimulation assay for measurement of cell-mediated immune responses in pigs experimentally infected with Lawsonia intracellularis

The whole-blood interferon-gamma (IFN-γ) assay is a quantitative in vitro assay for a direct read-out of Ag-specific cell-mediated immune (CMI) responses to infectious diseases. The IFN-γ assay is robust in severe intracellular infections like Brucella or mycobacteria, but more difficult to evaluate for less severe or immunocompromising infections. Here we investigated the performance of the assay when recombinant co-stimulatory cytokines IL-12 and/or IL-18 were added along with Ag or PBS to cultures of whole-blood from pigs infected with Lawsonia intracellularis. In pigs recovering from a natural infection, addition of rIL-12 or rIL-18 alone increased the Ag-specific IFN-γ release while addition of both cytokines resulted in increased IFN-γ release also in PBS cultures. In analyses after experimental infections with L. intracellularis, significant increased levels of Ag-specific IFN-γ production were measured in Ag+rIL-18 cultures from infected pigs compared to the background response in PBS+rIL-18 control samples (p<0.01) or to Ag+rIL-18 cultures from non-inoculated control pigs (p<0.05). Flow cytometry identified two lymphocyte subsets as the Ag-specific IFN-γ producers. The highest IFN-γ production was by CD4(+)CD8(+) cells while a more numerous population of CD4(-)CD8(+) cells produced lower amounts of IFN-γ in response to rIL-18 and L. intracellularis Ag.     

Bayesian estimation of true between-herd and within-herd prevalence of Salmonella in Danish veal calves

Specialised veal producers that purchase and raise calves from several dairy herds are potentially at high risk of delivering Salmonella-infected animals to slaughter. However, the true prevalence of Salmonella infected veal producing herds and the prevalence of infected calves delivered to slaughter from infected herds are unknown in Denmark. Due to uncertainties about test sensitivity and specificity, these prevalences are not straightforward to assess. The objective of this study was to estimate the within-herd- and between-herd prevalence of Salmonella in veal calves delivered for slaughter to abattoirs in Denmark. Furthermore, it was investigated to which extent the estimates differed between a setup using both serological tests and faecal culture, compared to just serological tests, and whether the applied sampling scheme in the national surveillance programme in Denmark was sufficient to establish high posterior estimates of freedom from infection in individual herds. We used Bayesian analysis to avoid bias as a result of fixed test validity estimates. Serological test results from 753 animals and faecal culture from 1233 animals from 68 randomly selected Danish veal producing herds that delivered more than 100 calves to slaughter per year were used to estimate the prevalences and estimates of freedom from Salmonella. Serological test results of 7726 animals from 185 herds were used to compare the difference in prevalence estimates between serology alone vs. faecal culture combined with serology. We estimated that 34-57% of specialised veal producing herds were infected with Salmonella. Within the infected herds, 21-49% of the animals were infected. Few herds obtained high posterior estimates for the probability of freedom from infection given the collected data, with only six of 68 herds obtaining posterior probability of being infected less than 10%. Furthermore, this study indicated that serology is sufficiently sensitive and specific to be used for estimating the prevalence of Salmonella-infected specialised veal producing herds.