Equilibrative nucleoside transporter 3 deficiency perturbs lysosome function and macrophage homeostasis

Lysosomal storage diseases (LSDs) are a group of heterogeneous disorders caused by defects in lysosomal enzymes or transporters, resulting in accumulation of undegraded macromolecules or metabolites. Macrophage numbers are expanded in several LSDs, leading to histiocytosis of unknown pathophysiology. Here, we found that mice lacking the equilibrative nucleoside transporter 3 (ENT3) developed a spontaneous and progressive macrophage-dominated histiocytosis. In the absence of ENT3, defective apoptotic cell clearance led to lysosomal nucleoside buildup, elevated intralysosomal pH, and altered macrophage function. The macrophage accumulation was partly due to increased macrophage colony-stimulating factor and receptor expression and signaling secondary to the lysosomal defects. These studies suggest a cellular and molecular basis for the development of histiocytosis in several human syndromes associated with ENT3 mutations and potentially other LSDs.     

Sole ulcers in Finnish dairy cattle

The Finnish Healthy Hooves project was set up to determine the frequency of, and risk factors for various hoof lesions in Finnish dairy herds. Data were collected in the years 2003 and 2004. A large dataset of over 74,000 cow-level observations recorded by hoof trimmers was merged with production data from the Finnish Agricultural Data Processing Centre Ltd. Ultimately, data from a single lactation from each of 16,792 cows in 703 herds were used for the analyses in this paper. Three-level hierarchical logistic models with hoof trimmer and farms (within hoof trimmer) as random effects were fit to data sets of tie stall (TS) and loose housing (LH) herds separately. The outcome of interest was the presence or absence of a sole ulcer in one or more legs of a cow during the lactation of interest.Cows examined once had a risk of sole ulcer 5.23% in tie stall herds and 7.58% in LH herds. As the number of examinations increased the odds of a diagnosis of sole ulcer increased substantially (2 and 3+ examinations had odds ratios (ORs) of 1.42 and 3.42 in TS herds and 2.77 and 6.89 in LH herds). Breed had a large effect on the risk of sole ulcer with Holsteins 2.89 times more likely to be affected than Ayrshires in TS herds and 2.94 times in LH herds. In TS herds, the presence of other hoof lesions such as haemorrhages (OR = 2.97), heel-horn erosions (OR = 2.10) and corkscrew claw (OR = 2.83) increased the risk of a sole ulcer developing. In LH herds, only haemorrhages (OR = 1.80) were a significant risk factor when parity was ≥2. In TS herds, use of mats (compared to hard flooring) significantly reduced the risk of sole ulcers (OR = 0.49). The effect of parity on the risk of sole ulcer was greatest when parity ≥4 but this effect was only significant in tie stalls (OR = 1.86).When analyses were restricted to cows with parity ≥2, similar results were obtained for the risk factors identified above. In addition, parity became highly significant in TS and LH (OR 2.31 and 2.23, respectively when parity was 4+). In TS herds, herd average milk production was significantly associated with a decrease risk of sole ulcer (OR = 1.28 per 1000 kg decrease) but there was no effect of production at the cow level (measured as deviation from the herd mean). No significant effects of production were observed in LH herds.     

Positional distribution of conjugated linoleic acid in triacylglycerol of Saccharomyces cerevisiae

Saccharomyces cerevisiae was cultivated in the presence of cis-9,trans-11 or trans-10,cis-12 isomers of free conjugated linoleic acid (CLA), and the effects of the isomers on the regioisomerisms of triacylglycerol (TAG) of the yeast were elucidated. Both isomers constituted about 34% of all fatty acids and increased drastically the number of different TAG species. Nearly all of the species contained CLA in at least one sn-position. In the most abundant species analyzed (20% of total species), the cis-9,trans-11 isomer appeared in combination with monounsaturated fatty acids (C16:1, C:18:1) whereas trans-10,cis-12 isomer was most frequently present with a medium chain fatty acid (C10:0 or C12:0) in the sn-2 position and C16:0 in one of the end positions (14% of total species). With either isomer, the amount of TAG species in which CLA encompassed all sn-positions was ca. 4%. Thus, S. cerevisiae can be used to produce edible single cell oil characterized by very heterogeneous distribution of CLA among the different TAG species.     

Microbially safe utilization of non-inactivated oats (Avena sativa L.) for production of conjugated linoleic acid

A microbially safe process for the enrichment of conjugated linoleic acid (CLA) in oats was developed. The process consists of hydrolysis of oat lipids by non-inactivated oat flour, followed by propionibacterium-catalyzed isomerization of the resulting free linoleic acid to CLA. The first stage was performed at water activity (a(w)) 0.7, where hydrolysis of triacylglycerols progressed efficiently without growth of the indigenous microflora of flour. Thereafter, the flour was incubated as a 5% (w/v) aqueous, sterilized slurry with Propionibacterium freudenreichii ssp. shermanii. The amount of CLA produced in 20 h was 11.5 mg/g dry matter corresponding to 116 mg/g lipids or 0.57 mg/mL slurry. The oat flour had also the capability to hydrolyze exogenous oils at a(w) 0.7. Sunflower oil, added to increase linoleic acid content in triacylglycerols 2.7-fold, was hydrolyzed rapidly. Isomerization of this oil-supplemented flour as a 5% slurry gave final CLA content of 22.3 mg/g dry matter after 50 h of fermentation, corresponding to 118 mg/g lipids or 1.14 mg/mL slurry. Storage stability of CLA in fermented oat slurries at 4 degrees C was good.     

Comparison of methods for measuring gap size distribution and canopy nonrandomness at Järvselja RAMI (RAdiation transfer Model Intercomparison) test sites

Methods for analyzing foliage nonrandomness by means of the TRAC instrument, digital hemispheric photography, and a gap fraction model are assessed at two RAMI (RAdiation transfer Model Intercomparison) mature stands in Järvselja, Estonia. The six different methods involve calculation of the canopy element clumping index, at scales coarser than that of a shoot. The major aim was to define the merits and limitations of the various methods. We conclude the gap size distribution and beyond-shoot clumping is very stable across the stands for the solar zenith angle range from 30° to 60°. Estimates based on the gap size distribution and the combination of gap size and logarithm methods performed the best while compared with an independent gap fraction model. We clarify the effect of the assumed leaf inclination angle distribution on gap size distribution and differences between estimates of beyond-shoot clumping. We show that the modified, gap-size distribution based method of Chen and Cihlar can provide reliable beyond-shoot clumping estimates without any a priori assumptions about the total gap fraction, segment length or the leaf inclination angle distribution. We also illustrate the changes in element clumping with measurement height. The compiled data extend the original parameter dataset to be used in the next phase of RAMI for different benchmark tests and reflectance modeling experiments, and contribute toward systematic validation efforts of radiative transfer models, operational algorithms, and field instruments, as promoted by the Committee on Earth Observation Satellites (CEOS).     

The effects of nitrogen fertilization on the ultrastructure of mesophyll cells of conifer needles in northern Finland

Cytological injuries in needles of Scots pine (Pinus sylvestris L.) and Norway spruce [Picea abies (L.) Karst.] were studied at light and electron microscope level in northern Finland. Needle samples were taken from fertilized (mainly with nitrogen) and nonfertilized trees during winter 1980. The results are discussed in relation to the severe climate and growth response of the trees.     

Serum cannabidiol,tetrahydrocannabinol (THC), and their native acid derivatives after transdermal application of a low-THC Cannabis sativa extract in beagles

Cannabinoids hold promise for treating health problems related to inflammation and chronic pain in dogs, in particular cannabidiol (CBD), and its native acid derivative cannabidiolic acid (CBDA). Information regarding systemic delivery of cannabinoids through transdermal routes is sparse. The purpose of this study was to determine pharmacokinetics of transdermal administration of a low-THC Cannabis sativa extract in healthy dogs. Six purpose-bred research beagles were treated with a transdermal CBD-CBDA-rich extract, and serum concentrations of CBD, CBDA, tetrahydrocannabinol (THC), and its acid derivative tetrahydrocannabinolic acid (THCA) were examined prior to and at the end of weeks 1 and 2. A 4 mg/kg dose of total cannabinoids twice daily resulted in appx 10 ng/ml of CBD, 21–32 ng/ml of CBDA, trace amounts of THCA, and unquantifiable amounts of THC in serum at the end of weeks 1 and 2 of treatment. Results showed that CBDA and THCA were absorbed better systemically than CBD or THC.