Thyroid sonography as an effective tool to discriminate between euthyroid sick and hypothyroid dogs

The diagnosis of canine hypothyroidism and its differentiation from euthyroid sick syndrome still is a major diagnostic challenge. In this study, ultrasonography was shown to be an effective tool for the investigation of thyroid gland diseases. Healthy control dogs (n = 87), dogs with euthyroid sick syndrome (n = 26), thyroglobulin autoantibody-positive (TgAA-positive, n = 30) hypothyroid dogs, and TgAA-negative (n = 23) hypothyroid dogs were examined by thyroid ultrasonography. Maximal cross sectional area (MCSA), thyroid volume, and echogenicity were measured. Statistical analysis identified highly significant (P < .001) differences between euthyroid and hypothyroid dogs both in thyroid volume and in MCSA, whereas no significant differences in thyroid size were detected between healthy euthyroid dogs and dogs with euthyroid sick syndrome. In euthyroid and euthyroid sick dogs, parenchymal echotexture was homogeneous and hyperechoic, whereas relative thyroid echogenicity of both TgAA-positive and TgAA-negative hypothyroid dogs was significantly lower (P < .001). When using arbitrarily chosen cutoff values for relative thyroid volume, MCSA, and echogenicity, thyroid volume especially was found to have highly specific predictive value for canine hypothyroidism. In summary, the data reveal that thyroid sonography is an effective ancillary diagnostic tool to differentiate between canine hypothyroidism and euthyroid sick syndrome.     

Productivity and quality of statice (Limonium sinuatum cv. Soiree Mix) and cockscomb (Celosia argentea cv. Chief Mix) under organic and inorganic fertilization regiments

Very little information is available on organic specialty cut flower production, especially fertilization requirements. In order to better understand organic fertilization requirements of two specialty cut flower crops, Limonium sinuatum and Celosia argentea, we initiated a field and greenhouse experiment to study the effect of compost (organic) and conventional (inorganic) fertilization treatments on the growth and productivity of these crops. Optimum yields in the field, expressed as fresh weight per plot, were achieved at compost applications of 98.8 t ha⁻¹ for both Limonium and Celosia. However, when number of stems, height of the stems, and the environmental impacts of such compost applications are considered we concluded that the optimal organic fertilizer amounts were 12.4 and 24.7 t ha⁻¹, for Limonium and Celosia, respectively. Limonium and Celosia plants in the greenhouse experiment were fertilized with 100, 200, 300, and 400 mg L⁻¹ nitrogen, combined with 0, 5, 10, 20, 40, and 60 mg L⁻¹ phosphorus. The generation of response surfaces for total weight per pot, number of stems per pot, average weight per stem, and average stem length were attempted for each species. The results showed that nitrogen did not significantly contribute to any of the models, except for Celosia average weight and length per stem models. Total weights per pot on the other hand showed both a linear and quadratic relationship over the range of phosphorus applications we tested. Maximum number of stems and total weight per pot were observed between 30 and 46 mg L⁻¹ P in both Limonium and Celosia. Our results suggest that organic fertilizer recommendations, in the form of animal manure composts should be based on phosphorus content of the compost rather than nitrogen content especially for soils high in initial phosphorous content.     

Histology-guided high-resolution AP-SMALDI mass spectrometry imaging of wheat-<Emphasis Type="Italic">Fusarium graminearum</Emphasis> interaction at the root–shoot junction

Background

Fungal pathogens like Fusarium graminearum can cause severe yield losses and mycotoxin contamination of food and feed worldwide. We recently showed its ability to systemically colonize wheat via root infection. However, the molecular response of wheat to Fusarium root rot (FRR) infection and systemic spread is still unknown. As a molecular camera, mass spectrometry (MS) imaging combines label-free and multiplex metabolite profiling with histopathology.

Results

Atmospheric-pressure (AP)-SMALDI-MS imaging was combined with optical microscopy to study wheat-F. graminearum interaction at the root–shoot junction, which is a crucial line of defense against a pathogen that can invade all distal plant parts. To scope the functional, temporal and local aspects of FRR disease spread, metabolic changes were simultaneous visualized in diseased and healthy stem bases of the resistant cultivar Florence-Aurore at 10, 14 and 21 days after root inoculation. Histological information was used to identify disease relevant tissues and to assist the interpretation of molecular images. Detected mycotoxin compounds secreted by F. graminearum showed a route of stem infection that was consistent with observations made by microscopy. The outer epidermis and vasculature of leaf sheath were, at different disease stages, identified as prominent sites of pathogen migration and wheat protection. Wheat metabolites mapped to these relatively small tissues indicated cell wall strengthening and antifungal activity as direct defenses as well as conservation in the wheat reactions to F. graminearum diseases that affect different plant organs.

Conclusions

AP-SMALDI-MS imaging at high spatial resolution is a versatile technique that can be applied to basic and applied aspects of agricultural research. Combining the technology with optical microscopy was found to be a powerful tool to gain in-depth information on almost unknown crop disease. Moreover, the approach allowed studying metabolism at the host–pathogen interface. The results provide important hints to an understanding of the complex spatio-temporal organization of plant resistance. Defense-on-demand responses to pathogen ingress were found, which provide opportunities for future research towards an improved resistance that does not negatively impact yield development in the field by saving plant resources and, moreover, may control different Fusarium diseases.

     

Characterization of mast cell numbers and subtypes in biopsies from the gastrointestinal tract of dogs with lymphocytic-plasmacytic or eosinophilic gastroenterocolitis

It has been suggested but not proven that hypersensitivity type I reactions are involved in the pathogenesis of canine inflammatory bowel disease (IBD). The main effector cells in type I hypersensitivity reactions are mast cells (MCs). Canine MCs, as human MCs, can be subdivided into three subtypes according to their content of mast cell-specific proteases: tryptase (MC_T), chymase (MC_C), or tryptase and chymase bearing MCs (MC_TC). In this study, numbers and subsets of mast cells were investigated in biopsies from the gastrointestinal tract of dogs with histopathologically confirmed lymphocytic-plasmacytic enteritis (LPE) (n = 4), lymphocytic-plasmacytic colitis (LPC) (n = 1) and eosinophilic gastroenterocolitis (EGE) (n = 11). Paraffin sections of formalin-fixed samples from the stomach, small intestine (duodenum, jejunum, ileum) and colon were stained by using a metachromatic staining method (kresylecht-violet; KEV) and a combined enzyme histochemical and immunohistochemical technique for chymase and tryptase. Additionally, immunohistochemistry with antibodies against T cells (CD3), macrophages (myeloid/histiocyte antigen) and IgA, IgG and IgM bearing cells was conducted. Quantitative evaluation of mast cells and semiquantitative scoring of immunohistochemically stained cells were performed. Between the two histopathologically defined groups clear differences concerning mast cell numbers were detected. In most affected intestinal tissue locations of dogs with LPE/LPC a decrease in metachromatically (kresylecht-violet) stained granule-containing MCs and immunohistochemically stained MC_T,C,TC was found. This reduction could be due to mast cell degranulation, a T helper cell 1 dominated reaction pattern or a “thinning out” due to increasing T cells, IgA and IgG bearing cells. Dogs with EGE displayed higher variability in mast cell numbers but most of the affected large and small intestinal locations had increased numbers of MCs. In these cases, T cells, IgA bearing cells and macrophages also increased. Increased numbers of MCs and eosinophils seen in the intestinal mucosa of dogs with EGE could indicate the presence of a type I hypersensitivity reaction (T helper cell 2 pattern) in response to dietary antigens. Changes in cell numbers occurred also in unaffected locations of dogs with LPE/LPC and EGE which showed reduced MC_T,C,TC, increased KEV positive cells and partially increased leucocytes and macrophages.     

Role of Plasmalemma H+ ATPase in Sugar Retention by Roots of Intact Maize and Field Bean Plants

Net release and net uptake of sugars by roots of intact maize (Zea mays cv. Blizzard) and field bean (Vicia faba L. cv. Alfred) were studied at micromolar external sugar concentrations that are relevant to the rhizosphere. Besides various sugars not further characterized there was net release of glucose, fructose, sucrose, arabinose, ribose, and galactose. The net release of these sugars into the root medium (0.1 mM CaSO₄) was stimulated by the protonophore CCCP (10 μM), the sulfhydryl reagent NEM (300 μM), the specific inhibitor of plasmalemma H⁺ ATPase vanadate (0.5 mM), and by the inhibitor of the glucose carrier phlorizin (2 mM). Net uptake of glucose, fructose, and arabinose from 10 μM external concentrations was inhibited by these substances. Stimulation of net release and inhibition of net uptake was most pronounced for glucose. Sucrose added to the root medium was hydrolyzed by invertase activity leading to glucose and fructose uptake by roots. It is concluded that the retention of sugars by plant roots is not only determined by plasmalemma permeability but is also controlled by the H⁺ electrochemical gradient established by ATPase activity (retrieval mechanism). The proton gradient drives a sugar/H⁺ cotransport system that is selective for glucose but may also transport other sugars, particularly in the absence of glucose.     

Does H+ pumping by plasmalemma ATPase limit leaf growth of maize (Zea mays) during the first phase of salt stress?

In the first phase of salt stress, growth of plants is impaired mainly by osmotic stress. To elucidate the effect of NaCl salinity on elongation growth of maize leaves in the first phase of salt stress, we investigated the effect of NaCl on gene expression and activity of the plasmalemma H⁺ ATPase of elongating leaves of maize (Zea mays L.). Treatment of maize plants with 125 mM NaCl for 3 d decreased leaf growth relative to control plants (1 mM NaCl). Whereas H⁺ ATPase hydrolytic activity was unaffected, the ability of the H⁺ ATPase to establish a pH gradient was strongly reduced. Total mRNA of plasmalemma H⁺ ATPase was slightly increased. However, mRNA of the ATPase isoform MHA1 was significantly reduced and ATPase isoform MHA4 was strongly increased at the mRNA level. Synthesis of total H⁺ ATPase protein was unchanged as revealed by western blot. The results indicate that reduced pumping of H⁺ ATPase in leaf plasmalemma under salt stress may be caused by a switch to gene expression of the specific isoform MHA4, which shows inferior H⁺‐pumping efficiency in comparison to isoforms expressed under control conditions. We propose that reduced H⁺ pumping of plasmalemma H⁺ ATPase is involved in the reduction of leaf growth of maize during the first phase of salt stress.     

Cd translocation into generative organs of linseed (Linum usitatissimum L.)

Linseed is able to accumulate considerably high concentrations of Cd in generative organs, the dietary critical value of 0.3 μg Cd/g seed often being exceeded. Differences between genotypes of linseed in this respect, however, have been observed but the underlying mechanisms responsible for these differences are unknown. The aim of the present work was to identify these mechanisms by studying the Cd accumulation of the linseed genotypes Antares and McGregor which differ in their ability to accumulate Cd in the seeds. Cultivar Antares is a high and cv. McGregor a low Cd accumulator which was confirmed in a pot experiment. It was found that the differences between these genotypes were Cd-specific and were caused neither by single seed weight nor by Cd translocation into the shoot. The distribution pattern of Cd within mature capsules between the pericarp and the seeds differed from that of Ca which was used as a phloem-immobile reference ion. From these results we conclude that Cd was translocated from the pericarp into the seeds via the phloem. This conclusion was supported by direct Cd determination in collected phloem sap from linseed stems. As sources of seed-Cd we identified the pericarps of capsules and the leaves. The genotype differences concerning the Cd concentrations in the seeds may be explained in terms of differences in phloem translocation of Cd.     

A diarylquinoline drug active on the ATP synthase of Mycobacterium tuberculosis

The incidence of tuberculosis has been increasing substantially on a worldwide basis over the past decade, but no tuberculosis-specific drugs have been discovered in 40 years. We identified a diarylquinoline, R207910, that potently inhibits both drug-sensitive and drug-resistant Mycobacterium tuberculosis in vitro (minimum inhibitory concentration 0.06 mug/ml). In mice, R207910 exceeded the bactericidal activities of isoniazid and rifampin by at least 1 log unit. Substitution of drugs included in the World Health Organization's first-line tuberculosis treatment regimen (rifampin, isoniazid, and pyrazinamide) with R207910 accelerated bactericidal activity, leading to complete culture conversion after 2 months of treatment in some combinations. A single dose of R207910 inhibited mycobacterial growth for 1 week. Plasma levels associated with efficacy in mice were well tolerated in healthy human volunteers. Mutants selected in vitro suggest that the drug targets the proton pump of adenosine triphosphate (ATP) synthase.